key: cord-0010250-9xpa492r
authors: nan
title: Abstracts for the 9th Congress of the Asian Pacific Society of Respirology 10‐13 December, Hong Kong
date: 2004-11-25
journal: Respirology
DOI: 10.1111/j.1440-1843.2004.00673.x
sha: e11f22b4126bd4ab6d18efdc0db8d56d9f0df77b
doc_id: 10250
cord_uid: 9xpa492r

nan

Background The most important pathological feature of asthma is airway inflammation which leads to airway hyperresponsiveness. The oxidation process is contributing in the airway inflammation of asthmatic patients. In the present study, the effects of long and short term ascorbic acid (which has antioxidant effect) administration were studied on airway responsiveness of sensitized guinea pigs. Methods An experimental model of asthma was induced in guinea pigs by sensitization of animals with injection and inhalation of ovalbumin (OA) on two groups; one with ordinary drinking water (group 2) and another with ascorbic acid (AA) in their drinking water (group 3). The responses of tracheal chains of control animals (group 1) and both groups of sensitized guinea pigs (group 2 and 3, for all groups, n = 10) to cumulative concentrations of methacholine (M) were measured, and the effective concentration of M causing 50% of maximum response (EC 50 M) was obtained. The responses of tracheal chains to 0.1% OA, relative to contraction obtained by 10 mM methacholine, were also measured. The tracheal responses to methacholine and OA were measured on both nonincubated and incubated tissues with AA. Results The tracheal responses of group 2 were significantly greater than those of group 1 and group 3 to both OA and methacholine (P < 0.05 to P < 0.001). There were no significant differences in tracheal responses to OA and methacholine between group 1 and 3 animals. Incubation of tissues caused reduction of tracheal response to methacholine in all three groups of animals but only in group 3 this difference was statistically significant (P < 0.05). However, incubation of tissues did not change tracheal response to OA significantly. Conclusion These results showed that although short term administration of ascorbic acid had no major effect on tracheal responsiveness of sensitized animals, long term administration of ascorbic acid can lead to decrease in airway responsiveness of sensitized guinea pigs to both OA and methacholine.

Background Bronchial asthma is associated with overreaction of Th2 cells Ag85B DNA vaccine was effective in stimulating Th1 protective immunity in Turberculosis, PBMC from TB patients in response to Ag85B stimulation showed enhance Th1 response after anti-TB therapy. The effect of Ag85B on T-helper cells cytokines in PBMC from asthmatic is unknown. Methods PBMCs collected from 18 mite-sensitized asthmatic patients and 13 healthy control were stimulated with mites, Ag85B protein, supernatant of cultured 16HBE cells transfected with pMG-Ag85B alone, or the combination of mites plus Ag85B protein or plus Ag85B supernatant. After 96 hours incubation, levels of IL-5 and IFN-g were determined. Results In the asthmatic group, there was an significant increase of IL-5 level after stimulation with mites as compared with control. Level of IFN-g increased significantly by co-stimulation of mites plus Ag85B protein or plus the Ag85B supernatant in comparison with the mites alone (87.60 ± 36.45) pg/ml or (111.5 ± 15.9) pg/ml vs (58.8 ± 7.8) pg/ml, respectively, P < 0.05. There was no significant difference among groups in IL-5 level in those patients or either IL-5 or IFN-g levels in normal subjects. Conclusions Ag85B protein or the supernatant harvested from cultured 16HBE cells transfected with pMG-Ag85B could upregulated Th1 response in PBMC with allergen stimulation from asthmatic patients. The annual asthma death toll in the Tohoku district was surveyed from 1992 to 2001. The survey inquired whether physicians or pediatricians had cases of asthma death during the period of the previous year, and was conducted by sending postcards in every February. Then, details of each fatality were further examined by questionnaire. A total of 299 cases of fatal asthma attack (male: 186, female: 113) were compiled from the survey conducted over a period of 10 years. Middle-aged and elderly asthmatics over 50 years old accounted for two-thirds of all the cases. Among asthmatics younger than 50 years age, the death toll of patients in their first decade was the highest. 78% of the total cases were outpatients. 60% of them were in cardiopulmonary arrest and 89% were classified as severe exacerbation or worse upon their arrival at the hospital. 25.6% and 39.6% of the death patients were diagnosed as dead only within 2 hrs and 24 hrs after the onset of asthmatic attacks, respectively. Cases of duration of 1 to 10 years of asthma outnumbered others, whereas 16 cases (5.3%) died within 1 year after the onset of their asthma. 34% of the cases had been scored as severe asthma before the death, whereas 45% and 21% had been diagnosed as moderate and mild, respectively. The number of deaths from asthma attacks in the Tohoku district was clearly reduced over the past 5 years. The current annual death toll plummeted by 75% compared to a decade ago, when our annual survey was commenced. In Tohoku district, death from asthma attacks was surveyed from 1992 to 2001. After presence of death from asthma attacks during the previous year were confirmed by postcard at the beginning of every February, details of each case were asked by questionnaire. In total, 299 cases (male: 186, female: 113) were recognized as death from asthma attacks. Middle-aged and older asthmatics over 50 years accounted for 2/3 of the cases and the most death was recorded in asthmatics in the second decade of life under 50. Outpatients accounted for 78%. When they visited hospitals, 60% was in cardiopulmonary arrest and 89% was in more critical condition than severe attacks. About duration of attacks from the onset to their death, within 2 hrs accounted for 25.6%, within 1 day did for 39.6%, and over 1 day did for 34.8%. Although the most death was found in patients whose duration of asthma was between 1 and 10 years, 16 cases died within 1 year from the onset of their asthma. About severity during the past year, 34% had severe asthma, 45% moderate, and 21% mild. Number of death from asthma attacks in Tohoku district was clearly reduced over the past 5 years. The number was reduced to about one-fourth of that at the beginning of survey.

Background & Objective p38 Mitogen-activated protein kinase (MAPK) plays a pivotal role in the activation of immune responses. In this study we investigated the effects of a respirable p38a MAPK antisense oligonucleotide (p38a-ASO) on airway inflammation in a murine model of asthma. Methods The p38a MAPK knock-down effect of p38a-ASO was confirmed in a mouse T cell line (EL4) and a mouse macrophage line RAW264.7 using Western blotting. BALB/c mice were actively sensitized by i.p. injections of ovalbumin (OVA) for 18 days followed by aerosol OVA challenge for the next 3 days. Daily aerosolized p38a-ASO treatments (estimated doses of 0.3, 1.5 and 3 mg/kg) of the mice were given from day 14 to day 20. Mismatched ASO was used as a negative control. Results p38a MAPK ASO but not the mismatched ASO dose-dependently suppressed methacholine-induced airway hyperresponsiveness in sensitized mice measured by whole body plethysmography. The p38a MAPK ASO significantly (P < 0.05) reduced OVA-induced increases in total cell counts and eosinophils recovered in bronchoalveolar lavage (BAL) fluid. In addition, histological studies reveal that p38a-ASO markedly inhibited OVA-induced inflammatory cell infiltration into the lungs (H&E stain) and mucus hypersecretion in the airway epithelium (PAS stain). The p38a-ASO dosedependently inhibited OVA-induced increase in Th2 cytokine levels in BAL fluid. Further, p38a MAPK ASO significantly (P < 0.05) abated the blood levels of total IgE, OVA-specific IgE and OVA-specific IgG 1 in sensitized mice as compared to the mismatched ASO. Conclusion Taken together, respirable p38a MAPK ASO may have therapeutic potential for the treatment of asthma. (This work was supported by a NMRC grant NMRC/0433/2000.)

H-M PIAO 1 Background The mechanism of goblet cell hyperplasia in the airway of asthma is incompletely understood. We examined the role of histamine in goblet cell hyperplasia using histamine deficient mice (HDC KO mouse) in which HDC gene was disrupted. Method Wild and HDC KO C57bl/6 mice were sensitized with ovalbumin (OVA). After two-week exposure to OVA, lungs were excised and goblet cell hyperplasia was evaluated histologically. Bronchoalveolar lavage fluid (BALF) was collected and cell differentials were calculated. RNA was extracted from lung tissues and Gob-5 mRNA was analyzed by the quantitative RT-PCR. Result While the total number of cells in BALF was significantly higher in KO-mice than in wild mice (28.2 ¥ 104 ± 3.4 vs 62.0 ¥ 104 ± 5.6 ¥ 104; mean ± SEM), the ratio of eosinophils was lower in KO-mice than in wild mice (91 ± 8% vs 42 ± 7%). The mRNA level of Gob-5 was greatly increased in KO mouse compared to wild mice (108 ± 35 vs 182 ± 61; Gob-5/GAPDH). The ratio of the goblet cell in the airway epithelium was markedly increased in KO-mice compared to wild mice. Conclusion It was suggested that histamine plays a role in goblet cell hyperplasia of airway epithelium in allergic airway inflammation.

Background With respect to patient education in the successful management of asthma, the impact of clinical practice guidelines in the local tertiary care setting is largely unknown. Objectives The aims of this study were to audit the level of knowledge of adult asthmatic outpatients at a tertiary care hospital, to determine the sources of asthma information and to determine variables associated with poor asthma knowledge. Methods We conducted a prospective, cross sectional survey on 94 asthmatic subjects using an interviewer-administered questionnaire. Results Of the 94 subjects, 39.4% were ignorant of the inflammatory nature of asthma while 56.4% did not understand the role of prednisolone in acute exacerbation of asthma. Only 17.0% reported having a written action plan. Lower educational level and older age were significantly associated with lower asthma knowledge scores. The doctor was the main source of asthma information. Asthma knowledge scores were significantly higher among those who named the doctor, pamphlets, newspapers, internet and books as a source of asthma information. In multiple linear regression analysis, educational level, having doctors and newspapers as a source of asthma information were independently associated with asthma knowledge scores. Conclusions Our study demonstrates that a large number of asthmatics have poor understanding of some aspects of their disease and have no written asthma action plan. Asthmatics with lower educational level have poor asthma knowledge and should be specifically targeted in asthma education programmes.

YJ CHOI, V SAZONOV KOCEVAR, L RADICAN Worldwide Outcomes Research, Merck & Co., Inc. USA Background and Objective Evidence from Western countries suggests that asthma is associated with high economic burden and impact on patients' and families' quality of life. We reviewed currently available literature to evaluate economic burden of asthma in adult patients from Asia. Methods Structured literature review searched available data between 1990 and 2004 using Medline Database, World Wide Web, abstract books from recent academic conferences and available reports. The following key words were used alone or in combination: asthma, allergic rhinitis, prevalence, resource use, cost, quality of life and functioning, and specific country names. Results Asthma-related resource use in Asia is mainly driven by the use of hospital and emergency services, as on average 40% but up to 86% of patients with asthma experienced hospitalization or required emergency assistance during any given one year period. The latter figure may be even higher in case of the patients with more severe asthma. Despite development and introduction of novel therapies, trends in asthma-related hospitalizations did not decline consistently over the past 2 decades mainly because of inconsistent use of available therapies and potentially due to patient aversion to inhaled treatment options. In addition, concomitant allergic rhinitis, which has been shown to worsen outcomes in patients with asthma, appears to be highly associated with asthma in Asia as well. Prevalence of AR among asthmatics was reported as 38.2% in all patients, and 10.1 ~74.4% in adults. This may further increase economic burden of asthma in the region. Conclusion This review shows that evidence on high economic burden of asthma in Asia is available, though may be of variable quality. Despite this it further emphasizes the need for improvement of current asthma control and treatment in Asia. Special attention should be devoted to patients with asthma and comorbid disease such as allergic rhinitis, as preliminary evidence suggests added burden in this population.

WP LIAO, H ZHU, J ZHAO, HPJ CHAN, WSF WONG Department of Pharmacology, National University of Singapore, Singapore

Background The inflammatory and remodeling processes that underlie asthma result from a highly complex interaction between various cell types and various cytokines. Apart from inflammatory cells, resident cells such as fibroblasts are also proposed to play critical roles in the switch from acute inflammation to adaptive immunity and tissue repair. Although asthma is perceived as a disease with a particular profile of Th2 cytokines, increasing evidence indicates that TNF-a, a classical Th1 cytokine, is also associated with the inflammatory response that characterize human asthma. Methods Total proteins from both cell lysates and supernatants between normal human lung fibroblasts (NHLF) and TNF-a induced NHLF were separated by two-dimensional (2-DE) gel electrophoresis, and proteins with significant changes were subsequently identified by MALDI-TOF mass spectrometry. RT-PCR was further applied to confirm the proteomics findings.

Results Several proteins were found to be markedly induced by TNF-a. These included some cell protective proteins: antiviral MxA GTPase, ubiquitin-like protein ISG-15, serine proteinase inhibitor PAI-2, and antioxidant protein MnSOD; and two intracellular enzymes for collagen modification: prolyl hydroxylase (PH) a subunit and lysyl hydroxylase 2, isoform a (LH2a). In addition, some proteins with unclear functions were found to be significantly down-regulated. Conclusion These results not only demonstrate that proteomics is an efficient way to study the underlying molecular mechanisms after cytokine stimulation, but also indicate that Th1 cytokine TNF-a, which is increased in asthmatic airway, can induce major alterations in global protein expression in NHLF leading to airway inflammation and remodeling. (Supported by a grant BMRC 01/1/21/17/046 by the Biomedical Research Council in Singapore.) Background and Objectives With increasing need to use limited resources effectively, study reports on burden of asthma in Asia are gradually growing both in terms of numbers and scope. We therefore reviewed and synthesized currently available data on resource use, school-absence, functioning and quality of life among pediatric patients with asthma from Asia. Methods Structured literature review searched available data between 1990 and 2004 using Medline Database, World Wide Web, abstract books from recent academic conferences and available reports. The following key words were used alone or in combination: asthma, child, resource use (i.e. hospitalizations), cost, school absence, functioning, quality of life and specific country names. Results Similar to adults, asthma-related resource use among children in Asia is mainly driven by the use of hospital and emergency services. Annually between 42.6-44.7% of children neither confirmed or suspected asthma visited asthma clinics/emergency departments (ED) in Hong Kong and China, where 3.5% of asthmatic children were also hospitalized due to an asthma attack. Similarly, asthma was the 2 nd most common reason for discharge and the 2 nd leading condition seen by general physicians and government polyclinics in Singapore during late 80s and early 90s. As seen in other regions, younger children (<5 years) were more than 2-times more likely hospitalized compared to older children (>5 years) and were thus responsible for about 60 ~80% of all asthmatic admission. School-loss, impaired functioning and impact on quality of life were also significant in young asthmatics. Conclusion Across Asia, there are currently few studies of resource use and general burden of asthma in pediatric patients. However, they consistently show higher impact on younger children (<5 years) and high reliance on emergency and hospital care for asthma. There appears to be room for improvement in terms of treatment options in order to reduce reliance on secondary care for asthma which will likely result in decreased economic burden on societies.

Objectives To evaluate the severity of asthma and its management at primary healthcare clinics in Melaka Tengah district of Malaysia. Methods A cross-sectional study with systematic sampling of asthma patients from August to October 2003 involving all primary healthcare clinics. Results A total of 207 patients were enrolled with mean age of 49 years and mean duration of asthma of 15.5 years. Malay ethnic group formed the majority (65.7%) and 54.1% were women. Most patients had severe persistent asthma (41.5%) and they were in older age group as well. Inhaled salbutamol sulfate was the most widely used quickrelieving medication (51.2%) whereas inhaled beclomethasone dipropionate was the commonest long-term preventive medication (35.7%) followed by sustained-release theophylline (8.7%) and inhaled budesonide (6.3%). Over half (55.2%) of the persistent asthma patients did not receive any inhaled corticosteroids therapy. Among those patients receiving inhaled corticosteroids, the median dosage was only 200 mg daily (min = 100 mg, max = 600 mg). Of the moderate and severe persistent asthma patients without any inhaled corticosteroids therapy, most of them were prescribed with regular oral short-acting bronchodilator (salbutamol, terbutaline and theophylline) acting as 'preventive treatment'. Inhaled corticosteroids were less frequently prescribed to patients with severe persistent asthma (39.5%) compared with mild persistent asthma (65.1%). Conclusion Most patients had severe persistent asthma and the asthma management was sub-optimal in which underuse of preventive medications is wide spread especially among those with severe persistent asthma. 1 

Background International guidelines recommend a range of clinical tests to confirm the diagnosis of asthma. These focus largely on identifying variable airflow obstruction and responses to bronchodilator or corticosteroid. More recently, the use of exhaled nitric oxide (FE NO ) measurements and induced sputum analysis to assess airway inflammation have been highlighted. However, to date, no systematic comparisons to confirm the diagnostic utility of each of these methods have been carried out. Methods We investigated 47 consecutive patients who were referred by their general practitioner with symptoms suggestive of asthma using a comprehensive fixed sequence of investigations and diagnostic accuracies were calculated. Results Mean FE NO levels were significantly higher in the asthmatic (n = 17) compared to the non-asthmatic (n = 30) patients (52.0 ppb (95% CI: 35.8, 68.2) and 15.7 ppb (95% CI: 11.1, 20.3) respectively, P < 0.001). Sensitivities and specificities respectively for a diagnosis of asthma were as follows; > 20% peak flow variation (0%, 100%); >15% peak flow response to steroid (24%, 100%); FEV 1 < 80% predicted (29%, 100%); FEV 1 /FVC ratio <70% (35%, 100%); >15% FEV 1 response to steroid (12%, 100%); induced sputum eosinophils analysis (86%, 88%); FE NO measurement (88%, 79%). Overall, the diagnostic accuracy based on comparisons of receiver operated characteristic (ROC) curves was significantly greater for FE NO and sputum eosinophils. Conclusion FE NO measurements and induced sputum analysis are superior to conventional approaches as diagnostic tools in asthma, with the advantage for FE NO that the test is quick and easy to perform. Supported by Otago Medical Research Foundation. A.D.S. is a GSK research fellow. Backgroud Airway smooth muscle proliferation plays an important part in airway remodeling in asthma. It is crucial to study the intracellular signal pathway in the airway smooth muscle cells (ASMCs) proliferation for understanding the mechanism of asthma and developing potential therapies. Protein kinase C (PKC) participates in the proliferation of many sorts of cells, but currently further study of PKC signal transduction in regulating ASMCs proliferation and the downstream of PKC activation is rarely found both home and abroad. Nuclear factor-kB (NF-kB), an important transduction factor, also assisting in proliferation of many cells, is mainly studied in vascular smooth muscle cells, and takes the part of downstream common factor in a great many signal transduction pathways. However, there has been no report whether NF-kB contributes to regulating ASMCs proliferation and whether it is the downstream factor of activated PKC or not. Methods As ASMCs from asthmatic patients is difficult to obtained, investigators abroad etablished a model for studying the biological behavior of asthmatic ASMCs by using serum from asthmatic patients to passively sensitized non-asthmatic human airway smooth muscle cells (HASMCs). In our experiment, 10% asthmatic patients' serum was prepared, as well as 10% non-asthmatic human serum as control. PKC agonist Phorbol 12-myristate 13-acetate (PMA), PKC inhibitor Ro-31-8220 and NF-kB inhibitor pyrrolidine dithiocarbamate (PDTC) were used to treat these cells. The NF-kB activation was detected by NF-kBp65 immunofluorescence staining and electrophoretic mobility shift assay (EMSA). The expression of inhibitory protein kBa (I-kBa) was detected by Western blot. The proliferation of HASMCs was examined by Flow Cytometry (FCM), MTT colorimetric assay and proliferating cell nuclear antigen (PCNA) immunofluorescence staining. The aim was to explore the contribution of NF-kB to the proliferation of passively sensitized HASMCs, and whether NF-kB was the downstream factor of activated PKC or not. Results According to results of the experiments, it was found that: (1) The proliferation of passively sensitized non-asthmatic HASMCs, expression of NF-kBp65 in nucleus and the combination of NF-kB with DNA were significantly increased, compared with those treated with controlled serum (P < 0.05); wheras when treated with 100 mmol/L PDTC, the proliferation of passively sensitized HASMCs, expression of NF-kBp65 and the combination of NF-kB with DNA were significantly decreased, only compared with passively sensitized non-asthmatic HASMCs (P < 0.05).

(2) I-kBa was degraded 15 minutes after HASMCs stimulated with asthmatic serum, and the degradation reached the peak after 60 minutes. After 90 minutes, the amount of I-kBa began to come back, and was returned to the level of control after 180 minutes. (3) After treated with 10 nmol/L PMA, the proliferation of HASMCs, expression of NF-kBp65 and the combination of NF-kB with DNA were significantly increased, only compared with passively sensitized HASMCs, HASMCs treated with both 10 nmol/L PMA and 100 mmol/L PDTC, HASMCs treated with 10 nmol/L PMA (P < 0.05). Conclusion All these findings indicated that NF-kB might take part in the proliferation of passively sensitized non-asthmatic HASMCs, in which there is PKC -NF-kB signal pathway. To block this signal transduction pathway on various points is significant to suppress asthmatic ASMCs proliferation and relieve airway remodeling.

Background In vitro studies show that single nucleotide polymorphisms (SNPs) of the beta2-adrenoceptor (ADRB2) alter receptor function. Clinical studies show that the Arg-16 variant is associated with increased response to single doses of beta-agonist, and, paradoxically, with decreased lung function during regular beta2-agonist use. The importance of ADRB2 haplotypes (combinations of SNPs) has received only limited attention. Our aim was to compare acute bronchodilator responsiveness for the five commonest ADRB2 haplotypes in patients with asthma. Methods 154 patients underwent prospective DNA analysis for ADRB2 haplotype (2/2, 2/4, 2/6, 4/4, 4/6, and 'other'). Inhaled beta-agonists were withdrawn completely and ipratropium was substituted as 'reliever'. Inhaled corticosteroids were withdrawn until 'loss of control' or for three weeks maximum. Spirometry was measured before and 20 minutes after inhaled salbutamol (2.5 mg nebulised) Results There were 28, 49, 25, 17, 21 and 14 patients harbouring the 2/2, 2/4, 2/6, 4/4, 4/6 and 'other' haplotypes respectively. The mean pre-bronchodilator FEV 1 (all) was 2.69 litres (SD 0.98), 81.6% predicted (SD 23.0%). The mean postbronchodilator FEV 1 (all) was 3.00 litres (SD 0.98), an increase of 13.9% (SD 11.9). For individual haplotypes, the % reversibility was: 2/2: 15.9 (10.6); 2/4: 11.2% (8.3); 2/6: 15.9% (15.3); 4/4: 17.6% (16.0); 4/6: 13.3% (12.5). Analysis of co-variance with baseline FEV 1 as co-variate did not reveal any significant relationship between ADRB2 haplotype and bronchodilator response. Conclusion In contrast to previous data, we were unable to confirm that ADRB2 haplotype is an important determinant of bronchodilator response in asthma. This study was funded by the Health Research Council of New Zealand.

Background Myofibroblasts, the main feature of which is represented by an important contractile apparatus similar to that of smooth muscle and in particular by the neo-expression of a-smooth muscle actin (a-SMA), have been thought to participate in airway remodeling in asthma. Bronchial epithelium with injury has been demonstrated to be responsible for the increase in levels of pro-inflammatory mediators. Thus we hypothesize that injured airway epithelial cells (AECs) might be driving sub-epithelial fibroblasts to differentiate towards myofibroblasts, through which, injured AECs play a critical role in the development of airway hyperreactivity (AHR). Methods Human primary cultured airway subepithelial fibroblasts (HPAFs) were co-cultured with human bronchial epithelial cells (16HBE-14o, HBECs) treated with lipopolysaccharide (LPS) for 12 hrs prior to co-culture followed by being scratched. The procedure was also performed in the presence or absence of ET-1 receptor A inhibitor (BQ123), anti-TGFb-1 neutralized antibody and BQ123 plus anti-TGFb-1 respectively. Following that, the HPAFs were subjected to the detection of alpha-SMA expression and cells proliferation by using western blotting, immunostaining and Brdu incorporation respectively. In addition, using MAPK signal pathways specific inhibitors SB203580, PD98058 and SP600125, the activity of main members of MAPK family including p38 kinase, ERK 1/2 and JNK in the processes was also tested for evaluating the role of MAPK pathways in the induction of alpha-SMA. To investigate the source of the increased ET-1 in the co-culture, the interaction between matrix metalloproteinas (MMPs) and ET-1 was studied by using cell transfection with anti-endothelin converting enzyme (anti-ECE) mRNA expression plasmid (P-anti-ECE) and gelatin zymography. Results HBECs treated with LPS plus mechanical injury induced a-SMA expression and accelerated proliferation in HPAFs. Addition of the both of BQ123 and anti-TGFb-1 blocked the induction completely. The co-culture with injured HBECs induced the production of Phosphorated p38 and ERK 1/2 in the HPAFs, but not in those HPAFs with normal 16 HBE. SB203580 and PD98059 abolished the induction of a-SMA expression in the HPAFs co-cultured with the injured HBECs. Both of MMP-2 and MMP9 released from the injured HBECs transfected with blank vector into supernatants were increased significantly, compared to normal control and the HBECs transfected with P-anti-ECE. Conclusion Our results suggest that Myofibroblasts transdifferentiation was induced in the subepithelial fibroblasts living with injury HBE cells, leading to the occurrence of AHR. The increase in levels of ET-1 and TGF associated with injured airway epithelium was contributing to the transdifferentiation through MARK signal pathways of P38 and ERK 1/2. Background The recent identification of new costimulatory pathways with specialized effector activities has afforded novel, prospective therapeutics for a variety of immunologic disorders. The inducible costimulator (ICOS)/B7-homlog 2 (B7-H2) has received attention based on its putative role in T cell effector function. The Requirement of ICOS/B7-H2 costimulatory pathway for asthma is controversial. Methods Female BALB/c mice sensitized with ovalbumin were administered with recombinant B7-H2 fusion protein 50 mg before the first challenge. The lungs were lavaged and removed 48 hours after the last challenge. The nucleated cells in bronchoalveolar lavage fluid (BALF) were counted and morphologically differentiated. The levels of IL-5 and INF-g in the BALF were determined by enzyme-linked immunosorbent assay (ELISA). The routine histologic analysis was performed. Results In recombinant B7-H2 group (n = 6), the pathologic change of airway inflammation was exacerbated, and the number of nucleated cells [(48.08 ± 4.50) ¥ 10 6 /L], eosinophils [(2.91 ± 0.20) ¥ 10 6 /L] and lymphocytes [(24.87 ± 0.94) ¥ 10 6 /L] in BALF was significantly increased as compared with asthma control group [(29.63 ± 5.22) ¥ 10 6 /L, (0.86 ± 0.10) ¥ 10 6 /L, (10.50 ± 0.55) ¥ 10 6 /L, respectively] (P < 0.05). The levels of IL-5 [(536.40 ± 112.30) ng/L] in the BALF from recombinant B7-H2 group were significantly higher than those in asthma control group [(379.84 ± 73.02) ng/L] (P < 0.05). INF-g levels [(100.20 ± 15.92) ng/L] in the BALF from recombinant B7-H2 group showed no differences as compare to that from the asthmatic control group [(98.73 ± 16.41) ng/L] (P > 0.05). Conclusion The costimulatory signals provided by B7-H2 and its receptor, ICOS, may be required for airway inflammation through regulating Th2 response in a murine model of allergic asthma.

TH ONG, KL TAN, P ENG Department of Respiratory and Critical Care medicine, Singapore General Hospital, Singapore

Purpose In Singapore, a National Program for asthma control has been set up since 2002, enrolling high-risk asthmatics and providing subsidized medication, a structured education program, and more rigorous tracing of defaulters. We sought to examine the impact of smoking among asthmatics enrolled in this program. Methods Data was extracted from a database of asthmatics from a single center in a nation-wide program for asthma control. Results There were a total of 438 patients enrolled as of October 2003. Among these, 109 (24.9%) reported a history of ever having smoked. 52 (11.9%) were current smokers, a figure similar to the national average of 13.8%. Current smokers were significantly younger (median age 38 years vs. 47 years in nonsmokers, P < 0.05), more likely to be male (76.9% vs 71.5%, P < 0.005) and reported a shorter duration of asthma (14.7 years vs. 17.1 yrs, P = 0.02). There was no difference in FEV1, PEFR or severity of asathma according to GINA classification. Interestingly, among those who had quit smoking, most (11/58 or 18.9%) reported that they had quit within the first year after their asthma was diagnosed. Conclusion Newly diagnosed asthma seems to be a significant impetus for smokers to quit smoking and efforts to encourage smoking cessation could be focussed on this group of patients. Smoking cessation should be an integral part of the asthma control program.

Pranlukast and zafirlukast are leukotriene D 4 (CysLT 1 ) receptor antagonists that are prescribed for asthma treatment. This study was addressed to evaluate the inhibitory potential of pranlukast and compare with that of zafirlukast, a known CYP2C9 inhibitor.

From the microsomal incubation studies in vitro, both pranlukast and zafirlukast showed no inhibition on CYP1A2-catalyzed phenacetin O-deethylation, CYP2C19-catalyzed S-mephenytoin 4¢-hydroxylation, CYP2D6-catalyzed dextromethorphan O-demethylation, CYP2E1-catalyzed chlorzoxazone 6-hydroxylation, CYP3A4-catalyzed midazolam 1hydroxylation. For the CYP2C9-catalyzed tolbutamide 4-methylhydroxylation, however, pranlukast and zafirlukast showed moderate inhibition with the estimated mean IC 50 s of 62.1 ± 12.3 mM and 25.5 ± 6.3 mM, respectively. From the full inhibition studies, both drugs appear to inhibit competitively the tolbutamide 4-methylhydroxylation with the estimated mean Ki values of 45.6 ± 4.0 mM and 16.3 ± 0.5 mM, respectively. These results suggest that pranlukast has also moderate inhibitory potential on CYP2C9-catalyzed tolbutamide 4-methylhydroxylation even though it appears to be not stronger than zafirlukast. Therefore, the inhibitory potential of pranlukast should be considered when CYP2C9 substrates with narrow therapeutic range (e.g., S-warfarin, phenytoin) are co-administered. Background Food is one of the causative agents associated with allergies. The true prevalence of food allergies unknown among population in India. Some studies have already been done to find out prevalence of food allergies in asthma.

Objective Objective was to determine the prevalence of immunoglobulin E-mediated food allergy against commonly consumed food in respiratory allergies.

Methods Cross sectional epidemiological studies were done in patients attending ENT and Allergy Centre. 500 Patients of respiratory allergy were screened to detect the senstivity of food allergens on the basis of questions. Patients clinical history was noted. Associated clinical examination was done. Associated problems were also noted down like rhinitis, sinusitis, utricaria or eczema. Relevent blood examination and other relevent tests were done. After confirmation of diagnosis of allergic asthma patients were taken for skin prick tests. Skin prick tests for environmental allergens (dusts, pollens, fungi and insects). In addition to environmental allergens food allergen tests were done for 40 to 50 food allergens depending on the food habits.

The results of various skin prick tests showed that patients having allergic asthma also show allergy to one or more food allergens. There is prevalence of 5-8% of food allergy. The common food allergens in our study are onion, mango, rice, mustard, citrus, yeast, bengal gram and kabuli chana and milk. Conclusion It is worth to do skin prick tests for food allergens in patients suffering from allergic asthma. 5-8% of patients suffering from allergic asthma showed excellent improvement after avoiding the food allergens. The remaining patients also showed good response after avoiding the food allergens along with the treatment of allergic asthma.

Background A diagnostic kit for influenza A and B was widely used in the Japanese medical scene about 2 years. We investigated the clinical relationships between influenza virus infection diagnosed by kit and exacerbation of asthma, including through the patients who had a medication at our hospital, Tokyo Emergency Room (ER) in two winter seasons.

Methods A total of 292 people suspected with influenza virus infection in winter 2002/3, and 739 people in winter 2003/4. All patients came to our hospital for cold, and received a checkup by the diagnostic kit for influenza A and B. Results We diagnosed 30 asthmatic patients among patients of influenza positive with the diagnostic kit. Among 30 patients, 27 people were diagnosed Influenza A, the other 3 patients were diagnosed Influenza B. We divided the influenza A patients into two groups; the cases accompanying with an acute exacerbation of asthma was group 1 (G-1): 12 patients (44%), and the cases without exacerbation was group 2 (G-2): 15 patients (56%). The average age of G-1 is higher than G-2. (mean age ± SE: G-1 51.1 ± 5.1 y, G-2: 28.0 ± SE2.9 y).There was numerically superior in female in G-1 (men : female = 1 : 11). We also divided asthma patients by the classification of JGL'2003 (Guidelines for the diagnosis and management of bronchial asthma by JAPANESE SOCIETY OF ALLERGOLOGY: JSA) based on the serious degree; step 3 and step 4 were predominant in G-1 (step 1: 25%, step 2: 8.3%, step 3: 33.3%, step 4: 25%): step 1 and step 2 were predominant in G-2 (step 1: 61.1%, step 2: 22.2%, step 3: 11.1%, step 4: 0%). There was no clinical exacerbation of asthma in patients with influenza B.

Conclusion We experienced the cases with acute exacerbation of asthma caused by influenza A virus infection. These asthma patients with exacerbation had a tendency to more serious than without exacerbation.

Introduction Asthma is major cause of morbidity in Hong Kong. Asthma self-management plan has been shown beneficial in reducing asthma morbidity, use of health services and even mortality in studies in many parts of the world. However, data on the effectiveness of such programme in Hong Kong are lacking. Study Objectives To evaluate the effectiveness of an asthma selfmanagement programme on asthma-related use of medical services, absenteeism from work and morbidity in patients admitted to the Queen Mary Hospital for acute asthma in Hong Kong. Method Patients were randomized into the intervention and the control group, and followed up for 12 months. The intervention group received the Asthma Self-management Program with a written self-action plan and usual care while the control group received only usual care. Both groups were given asthma education. Comparisons of outcomes at 12 months were made between the 2 groups with respect to the ratio of asthma related hospitalization, A&E visits, visits to general practitioners, days off work, asthma symptomatology, use of medications and lung function.

Results Forty patients were randomised into the intervention and 36 into the control group. Both groups have comparable baseline characteristics and similar rate of lost to follow up (~13%). At 12 month, patients in the intervention group were significantly less likely to have cumulative days off work or off school (RR = 0.23, 95% CI: 0.13 to 0.34) and unscheduled doctor visits (RR = 0.60, 95% CI: 0.30 to 1.00). There were no significant differences in the incidence of hospitalization, A&E visits, frequency of asthma symptoms and lung function between the two groups. Conclusion Asthma self-management plan is an acceptable and effective way of reducing asthma related absenteeism from work and unscheduled visits to doctors. It should be included in the management of adult patients with asthma in Hong Kong. Background Nebulized coticosteroid have a greater antiinflamatory potency and fewer systemic effects than intravenous or oral corticosteroid. However, their role in acute asthma has not been established. We prospectively investigated the efficacy and safety of nebulized corticosteroid in controlling severe acute asthma in the emergency department.

Methods The study was randomized clinical trial. Thirty eight subjects severe acute asthma treated with combination nebulized fluticasone 0.5 mg and salbutamol 2.5 mg, 38 subjects treated with methylprednisolone intravenous 125 mg and nebulized salbutamol 2.5 mg. We evaluated peak expiratory flow rate (PEFR) and symtom scores for six hours.

Result Subjects were 76 patien severe acute asthma. At the end of the study there was similar improvement PEFR in the two group. (r > 0.05).

Nebulized fluticasone goup showed early respons than methylprednisolone.

Conclusion The study shows that nebulized fluticasone is same as methylprednisolone intravenous in controlling acute asthma. We have known the important role of the peripheral nerve in the onset of asthma, Some Phenomenon has shown that psychological factor affected the onset of asthma, but up to now the role of the peripheral nerve in the onset of asthma has not been proved accurately. Our experiment investigates the influence of central nerve function on the onset of asthma. Methods Sixteen adult guinea pigs were divided into four groups of four cases in random: control group, saline group (NS), exhilarant stimulant group and inhibitor group. Injected the ovalbumin to abdominal cavity to establish the models of asthma guinea pigs, and then injected saline, aurintricarboxylic-acid (ATA) and MEK inhibitor (PD98059) into the lateral ventricle respectively. After that we observed the variations of praxiology, respiratory resistance and airway exudation. The experimental animal caused asthma by absorbing the ovalbumin aerosol, but the control group absorbed saline instead of the ovalbumin aerosol, and then 1-2 minutes later, we assay the respiratory resistance and respiratory rate respectively. In order to assay the airway exudation, we use Evinse Blue and measured the OD value 1 minute after guinea pigs inhaling ovalbumin aerosol.

Results (1) The values of respiratory resistance of four groups is as follows respectively: control group (0.11 ± 0.03) cmH 2 O; Ns group (0.74 ± 0.11) cmH 2 O; PD98059 group (0.42 ± 0.03) cmH 2 O; ATA group (1.45 ± 0.17) cmH 2 O. There were significant differences among the variations of each group. Between the NS group and the control group, t = 13.858, P = 0.01; between the PD98059 group and the control group, t = 7.020, P = 0.006; between the ATA group and the control group, t = 14.138, P = 0.001; between Ns group and PD98059 group, t = 20.239, P = 0.000; between Ns group and ATA group, t = 9.530, P = 0.002; between the PD98059 group and the ATA group, t = 16.895, P = 0.000.

(2) assay of the airway exudation:

We can detected EB in the airway of control group guinea pigs with the amount of (30 ± 8) ng/mg. NS group with the amount of (67 ± 25) ng/mg, which was higher than that of control group, t = 6.798, P = 0.007; PD98059 group with the amount of (45 ± 10) ng/mg, which was lower than that of control group, t = 3.367, P = 0.043; ATA group with the amount of (142 ± 46) ng/mg, which was significantly higher than that of NS group, t = 6.032, P = 0.009.

(3) variation of praxiology: there were significant difference between each group in the respiratory rate and reaction to stimulation. Conclusion The changes of central nerve system have an influence on the onset of asthma. This influence includes respiratory resistance, airway exudation and variation of praxiology.

LQ SONG 1 Background The increase of expression and activity of mice calciumactivated chloride channel 3 (mCLCA3) on airway goblet cells plays a key role in inducing mucus overproduction in asthmatic mice. The inhibition of mCLCA3 by active immunity should be a useful approach to prevent asthma. Methods We recombined the DNA vaccine based on xenogeneic homologous human CLCA1 (hCLCA1) and named it pSecTag2B/hCLCA1. BALB/c mice were vaccinated by i.m. once every two weeks with vaccine. Asthma was induced with ovalbumin in DNA vaccine group when antiserum was identified having the binding activity to extracellular domains (ED) of mCLCA3 by ELISA analysis. Other mice were divided into control-vector, control-saline and normal group. Asthma was also induced in the two control groups. The number of airway goblet cells was detected with periodic acid-Schiff staining, and lung MUC5AC mRNA level was detected by reverse transcription polymerase chain reaction assay. Results Antiserum of DNA vaccine group showed good binding activity to three mCLCA3 EDs, and the activity to N-terminal and C-terminal ED was stronger than middle ED. After asthma induction, the number of goblet cells in small bronchioles and MUC5AC mRNA level of lung in vaccine group were significantly lower than those in two control groups. There was no significant difference between vaccine group and normal group. Conclusion hCLCA1 DNA vaccine can induce mouse to produce serum antibody cross-binding to mCLCA3 itself, and thus airway mucus overproduction of asthmatic mouse is effectively inhibited. The mechanism of vaccine may be that mCLCA3 activity were blocked by antibody.

Background Bronchial asthma is characteristic of chronic inflammatory process in airway, and IL-4 and IL-13 secreted by Th2 cells play very important roles in the pathogenesis of asthma [1] .

Recently, it has been found that CD4+45RO+ T cell/memory T cell is a kind of dominating cell in asthma, for example, about 95 percent of total cells in bronchial walls of asthma are memory T cells [2] . The selective activation of CD4+45RO+ T cell can conduce assembling of eosinocytes, airway hyperreactivity and asthmatic attack [3] . Furthermore, more and more studies about asthmatic pathogenesis focus on costimulatory molecules now. Besides the main CD28-B7 pathway [4], CD152-B7 pathway has been paid more attention, which plays a negative regulatory role in proliferation of T cells and can inhibit the production of cytokines induced by allergens [5] . The object of our study is researching the effect of anti-human CD152 monoclonal antibody on secretion of IL-4 and IL-13 of CD4+45RO+ T cell in patients with atopic asthma. In our study, 16 subjects were recruited, including 8 patients with atopic asthma, who had medical history of mite allergy, positive bronchiectasis test, positive skin test about mite, positive mite-specific IgE in sera, and who had not been given glucocorticoid before drawn blood, and 8 healthy control ones. We drew 10 ml anticoagulated blood from every subject, and then got CD4+45RO+ T cell by immunomagnetic microbead abstraction, whose purity checked by flow cytometer was more than 95%. When we mixed cultured CD4+45RO+T cells with self B cells, we designed three groups: without stimulation, single mite extractive antigen stimulation and stimulation with mite extractive and antihuman CD152 antibody together. Then we collected supernatant of every sample after culturing 36 hours, and detected the concentration of IL-4 and IL-13 in supernatant by ELISA. Results 1. There was no difference between asthma and control in the production of IL-4 and IL-13, when CD4+45RO+T cells were cultured without stimulating (P > 0.05). 2. The concentration of IL-4 and IL-13 secreted by CD4+45RO+T cells increased on the asthmatic group after mite extractive stimulating (P < 0.05), and was higher than control group (P < 0.05), who had no change after mite extractive stimulating (P > 0.05). 3. The CD4+45RO+ T cells of control subjects secreted significantly less IL-4 and IL-13 when stimulated by mite extractive and antihuman CD152 antibody together than when stimulated by mite extractive singly (P < 0.005). But there was no significant change in IL-4 secretion of asthmatic patients' memory T cells after adding antihuman CD152 antibody (P > 0.05). There was slightly decrease in IL-13 secretion of asthmatic patients'memory T cells after adding antihuman CD152 (P < 0.05), but the percentage of decrease was less than control subjects (P < 0.05). Because CD4+45RO+ T cells of asthmatic patients were mite specific in our study, when they were stimulated with mite extractive once, they were activated and secreted more IL-4 and IL-13, as maybe play an important role in the pathogenesis of asthma. Our data show that antihuman CD152 antibody can inhibit the secretion of IL-4 and IL-13 in CD4+45RO+ T cell of control subjects significantly. It has been shown that CD152 plays an important role in regulating initiation and termination of T cell response in some studies. Especially at the later stage of immune reaction, CD152 can bind to B7 more efficiently than CD28, so that it can inhibit T cell entering S stage from G1 stage, inhibit the activity of IL-2 transcription factor, and cause downregulation and termination of T cell response. In our study, we adopted a small dose of antihuman CD152 antibody as stimulant, which can activate but not block CD152 on CD4+45RO+ T cells, open the CD152-B7 pathway, and inhibit secretion of IL-4 and IL-13 by memory T cells. But in asthmatic subjects, antihuman CD152 antibody can not downregulate IL-4 secretion, and the decrease of IL-13 is lower than control subjects. The reason for this result, we think is that: 1. CD152 on allergen-specific CD4+45RO+ T cell of asthmatic patients has been over activated (but still less activated than CD28), so it is not sensitive to small dose antihuman CD152 antibody. 2. There is some disfunction on CD152 on CD4+45RO+ T cell of asthmatic subjects, so the response to CD152 antibody is weak. Conclusions CD4+45RO+ T cell of patients with atopic asthma can secrete more IL-4 and IL-13, when touching the same allergen once. Antihuman CD152 antibody can activate CD152 on CD4+45RO+ T cell of healthy subjects, simulate the inhibition of CD152-B7 pathway, and then reduce the secretion of IL-4 and IL-13. But in asthmatic patients, the activation of antihuman CD152 antibody to CD152 is weaken. The results hint that CD152-B7 pathway or CD152 on CD4+45RO+ T cell of asthmatic patients is abnormal, as maybe can explain partly the increase of secretion of IL-4 and IL-13 in asthmatic patients. Background Increased activation of nuclear factor kB (NF-kB) might be the basis for chronicity of airway inflammation in asthma.

Objective To investigate the effect of arsenic trioxide (As 2 O 3 ) on activation of NF-kB and expression of IkBa in a murine model of asthma, and to explore its possible mechanism for antiinflammation. Methods Thirty-six BALB/c mice were randomly divided into the control group, the asthmatic group (subdivided into 1, 4, 12 and 24 h timepoints after the last airway challenge of ovalbumin), and the therapeutic group of As 2 O 3 (4 mg/kg). The characteristic recruitment of eosinophils (EOS) in bronchoalveolar lavage fluid (BALF), the pulmonary NF-kB activity and IkBa expression were detected by Diff-Quick staining, electrophoretic mobility shift assay (EMSA) and Western blot analysis, respectively. Results In the control group, the recruitment of EOS in BALF, pulmonary NF-kB activity and IkBa expression were (0.5 ± 0.2)%, 51.4 ± 4.5 and 0.80 ± 0.25, respectively. The asthmatic group had higher recruitment of EOS in BALF (P < 0.01) and pulmonary NF-kB activity (P < 0.01) and lower pulmonary IkBa expression (P < 0.01) than did the control group, respectively, in which NF-kB activation was increased within 1 h (P < 0.01), peaked by 4 h (P < 0.01), and dissipated by 12 h (P < 0.01) to 24 h (P < 0.05) after the last airway challenge. In contrast to the asthmatic group, the three parameters changed reversedly significantly (P < 0.01) after exposure to As 2 O 3 , and there was a significant negative correlation between either recruitment of EOS in BALF or pulmonary NF-kB activity and pulmonary IkBa expression (r = -0.82 and -0.94, P < 0.01). Conclusion It is a vital mechanism for As 2 O 3 to exert its multiple antiinflammation by inhibition of pulmonary NF-kB activation through induction of IkBa expression. Background Chronic obstructive pulmonary disease (COPD) is a complex disease which is influenced by genetic factors, environmental influences, and genotype-environment interactions. Smoking is the major causal factor in the development of the disease, While only 10-20% of chronic heavy cigarette smokers have respiratory symptoms, which suggests the presence of genetic susceptibility. Protease-antiprotease imblance is the mechinsm of COPD, MMP-9 has proteolytic activity against connective tissue proteins and it plays important roles in the destruction and remodeling of lung tissue of COPD. So we investigate the correlation between Matrix Metalloproteinase-9 gene polymorphism and chronic obstructive pulmonary disease susceptibility in Chinese population of Han nationality.

Methods Examine the frequency of polymorphic genotypes of the promoter of MMP-9 (-1562C/T) in 100 COPD patients and 98 healthy smoking by restriction fragment length polymorphism.

The frequencies of polymorphic genotypes in promoter of MMP-9 in COPD group were C/C 86%, C/T 14%, the frequencies of polymorphic genotypes in control group were C/C 98%, C/T 2%. There were significant difference between two group (P < 0.01). The difference in allele frequencies were also significantly between two group (C allele frequency: 93% vs 99, T allele frequency: 7% vs 1%, P < 0.05).

Conclusion The genetic polymorphism in promoter of MMP-9 gene is associated with the susceptibility to COPD in Chinese population of Han nationality. Background Pulmonary epithelial cells can act as 'effector' cells, synthesizing and releasing cytokines in response to inflammation of Chronic obstractive pulmonary disease (COPD). Cyclooxygenase-2 (COX-2), an inducible enzyme expressed in response to inflammatory cytokines, responsible for the synthesis of pro-inflammatory PGs such as PGE 2 . Statins are inhibitor of HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase, which is crucial in cholesterol biosynthesis. Resently, antiinflammatory effects of statins have been described. So, we suppose that atorvastatin, a kind of statin, affect the expression of COX-2 and production of PGE 2 , IL-6 in cultured human pulmonary epithelial cells.

Objective To assess the effect of atorvastatin on the expression of COX-2 and inflammatory cytokines in human epithelial cells (A549). Methods A549 cells were co-incubated in the same medium in presence of LPS for 12 h in different concentration of atorvastatin (0, 10, 15, 20 mM, respectively). After stimulation, supernatants were collected to measure the stable metabolite of prostaglandin E 2 (PGE 2 ), interleukin-6 (IL-6) using enzyme-linked immunosorbent assays (ELISA Chronic obstructive lung disease is one of the major causes of morbidity and mortality all over the world. Tobacco smoking is principal risk factor for COPD and traditional baking with wood and animal dung is another risk factor especially in non-smoking females in our country and some other places. We have studied reversibility of airflow limitation to salbutamol and oral steroid in female non-smoking bakers affected by COPD. Materials and methods Eighty-four non-smoking baker females presented with dyspnea, cough, sputum, and wheezing with FEV1/FVC% < 70 and without any interfering drug intake were selected with convenience method, and rapid bronchodilator response to salbutamol and delayed response to 30 mg prednisone for 2 weeks was assessed using ATS guideline for bronchodilator responsiveness (FEV1 or FVC ≥ 12% with 200 ml increase over baseline for salbutamol and FEV1 or FVC ≥ 20% for steroid). Sixtynine of 84 patients was selected for final report due to completion of the tests. Uncooperative patients or those unable to take steroid were excluded from study. Results Twenty-eight patients (40%) showed response to salbutamol. In remaining 34 patients only 6 showed >20% increase in FEV1 after 2 weeks of oral steroid. The overall response was about 50% considering both tests. The same picture was noted for FVC. Conclusion bronchodilator response in non-smoking female bakers to salbutamol and oral steroid is higher than we expect in common smoker's COPD and so asthmatic component may play a major role airflow limitation of these patients. Long-term effect of inhaled steroids has worth to be tried in this population of patients. Background Hypersecretion of mucus in airway is an important feature in chronic obstructive disease but effective treatment still lacks. In this study, we investigate the effect of dexamethasone on acrolein-induced MUC5AC expression in rat airways and whether it affects through signal transduction pathway mediated by nuclear factor-kB and protein kinase C. Methods Rat model of mucus hypersecretion of airways was established by exposing to 3.0 ppm acrolein fog 6 hours per day for 12 days. Animals were also injected intraperitoneally with dexamethasone (0.1 mg.kg -1 .d -1 or 0.5 mg.kg -1 .d -1 ) with or without exposing to acrolein fog. The mRNA of MUC5AC and the protein of MUC5AC, nuclear factor-kB, IkB-a and PKC in trachea of each animal were detected by RT-PCR and immunohistochemistry. The release of TNFa and IL-8 in BALF was measured by ELISA.

Results Exposure to acrolein for 6 days increased the expression of MUC5AC mRNA and its protein, the percentage of NF-kB positive nucleistained cells and PKC staining in trachea but decreased I-kB staining when compared with those in control (P < 0.01). The treatment with dexamethasone significantly reduced the expression of MUC5AC, PKC and the percentage of NF-kB positive nuclei-stained cells but increased expression of I-kB (P < 0.01). The increased release of TNFa and IL-8 in BALF after exposed to acrolein were also attenuated when treated with dexamethasone.

Conclusion Dexamethasone decreased NF-kB activation through upregulation of the expression of IkB-a, and reduce PKC expression, through which dexamethasone might reduce the expression of MUC5AC and be effected on over-expression of MUC5AC in rat airway exposed to acrolein fog. Background Airflow limitation is a major feature of severe COPD. One of the contributing mechanisms is airway narrowing that we already showed a newly developed cinema as dynamic imaging of high speed CT during quiet breathing in a patient with severe emphysematous lung in APSR 2003.

In that presentation, airways those diameters were around 3 mm were profoundly changed in cross-sectional area upto 75% decrease in expiratory phase. Since we observed airway narrowing not only in the small diameters airways but also in the trachea, in the present study, we analyzed and compared tracheal cross-sectional areas.

Methods Seven patients with severe COPD (%FEV1; 24.8 ± 4.0%), and 3 healthy subjects (age; 64.3 ± 3.5, and 64.1 ± 8.2 years, respectively) were studied. The chest was scanned using electron-beam computed tomography (Imatron C-150). CT images were captured during resting breathing with a series of twenty 100-msec images at an interval of 500 msec. The thickness of these slices was 3 mm. Each CT image was saved as a DICOM file, and was used to construct a digital video image using personal computer software.

In the cinema, the cross-sectional area of the trachea was maximum in inspiratory phase, and clearly decreased in expiratory phase in patients with severe COPD. The percentile decrease was statistically significant compared to the healthy subjects (27.5 ± 13.5% vs 7.6 ± 3.5%, respectively; P < 0.05). Conclusion These results suggest that even trachea as well as the airways in small diameter were compressed and/or collapsed in expiratory phase during quiet breathing in patients with severe emphysema. Background In previous work, we designed and tested questions to help identify COPD among two risk groups: 1) persons with a positive smoking history but no prior evidence of obstructive lung disease ('case-finding'), and 2) patients with prior evidence of obstructive lung disease ('differential diagnosis'). For these questionnaires, we now present a scoring system for use in primary care. Methods We developed separate scoring systems for each questionnaire. Scores were developed by weighting responses to each question based upon its regression coefficient from the final regression model. Two thresholds or 'cutpoints' for the scores were chosen to assign subjects to one of three zones of likelihood of having fixed obstruction: 1) High Likelihood (high predictive value of a positive test), 2) Intermediate Likelihood, or 3) Low Likelihood (high predictive value of a negative test).

The following shows how the questionnaires and scoring systems performed relative to spirometry:

Background The importance and the need of pulmonary rehabilitation that reduces dyspnoea, increases exercise capacity and improves the QOL of patient with chronic respiratory failure are widely recognized. These effects are expected additionally even in the patients who have already been reduced their symptoms with medication. To evaluate the influence of singing as one of the breathing conditionings upon patients heartrates and the oxygen saturation (SpO2), we made comparisons between singing, strech exercise and walking.

Methods 84 patients with chronic respiratory failure (COPD 64, bronchial asthma 12, interstitial pneumonia 5, bronchiectasis 1, old TB 2) were included, and they had attended pulmonary rehabilitation once to 17 times (mean 4.4 times). Patients heartrates and SpO2 were observed with percutaneous pulsoxymeter before and after the strech exercize, 6-min walking and singing in seated position. Background Chronic obstructive pulmonary disease (COPD) is a major public health concern in Asian countries since smoking prevalence rates are extremely high and continue to increase. Furthermore in the Asian-Pacific regions, which comprise both developed and developing countries, indoor air pollution from biomass fuel, utilized for cooking and to generate heat (heating) in poorly vented dwellings, is postulated to be another important risk factor for the development of COPD. Methods We conducted a questionnaire survey toward chest physicians or internists to investigate the awareness and consensus on the GOLD guideline as well as how COPD patients have been cared and managed in Asian region since the release of the GOLD guideline. The survey was conducted twice, in June 2001 and March 2004, in 9 countries including Japan, China, South Korea, Indonesia, Pakistan, Hong Kong, Taiwan, Thailand, and Philippines. A questionnaire was sent by mail in Japan while answered in a face-to-face setting in other countries.

Results Overall, about 84% of the responders were aware of the publication and updating of the GOLD guidelines. In 5 of 9 countries surveyed, COPD patients were cared and managed according to the combination of local and GOLD guidelines (43.1-71.1% of the responders in each country), or mainly according to the local guideline in China (45.2%), Hong Kong (53.3%) and Taiwan (48.1%) while the GOLD guideline was used mainly in Korea (50%). The majority of responders in each country (71.0-100%) answered that it is necessary to develop a local or regional guideline for best practice in the Asian-Pacific regions. Conclusion GOLD is successfully recognized and utilized in daily practice of the care and management of COPD in the Asian-Pacific regions as well as stimulated the development of local and regional guideline to optimize prevention, care, and management of COPD according to local characteristics. Background The purpose of this study was to analyze the coexistence of diastolic dysfunction in patients with advanced chronic obstructive pulmonary disease (COPD), in order to validate our clinical experience of high prevalence of diastolic dysfunction and the beneficial effects of intervention in these patients.

We studied a random sample of 21 consecutive patients of advanced COPD (GOLD stage III) diagnosed on the basis of pulmonary function tests, for evidence of coexistent diastolic dysfunction in 2-D echocardiography and doppler studies. None of them had documented cardiac illness or hypertension, and all had normal ECG.

The study population comprised of elderly males, aged between 53 and 83 years (median: 63 years). Ten patients had predominant emphysema (47.6%), 4 had predominant chronic bronchitis (19%), and the rest (n = 7) had mixed clinical pictures of both emphysema and chronic bronchitis (33.3%). The median FEV1 percent was 28. Overall, diastolic dysfunction was present in 15 patients (75%), of which eight patients (53.3%) had predominant emphysema, six patients had mixed disease (40%), and one patient had chronic bronchitis (6.7%). We found that patients with higher FEV1 had lower likelihood of having concomitant diastolic dysfunction (Odds Ratio: 0.18, 90% CI: 0.04-0.78), and that resting tachycardia was positively associated with diastolic dysfunction (OR: 9.75, 90% CI: 1.38-68.8). Conclusion Diastolic dysfunction is highly prevalent among advanced COPD patients. Low FEV1 and resting tachycardia are potential clinical markers for diastolic dysfunction among COPD patients. Background Measurement of health status is an important component to compliment physiologic measures in assessing the natural history and/or response to therapy in patients with COPD. There are relatively few reports of the relation of these measures in Japanese COPD patients. Methods Two clinical trials in the tiotropium development program in Japan (n = 193 and 147) afforded the opportunity to evaluate the association of health status, symptoms, and spirometry. Measured parameters included spirometry, respiratory symptoms, as well as patient impression (PI), physician evaluation (PE) and St. Geoge's Respiratory Questionnaire (SGRQ) (one study). Associations between the responses (i.e. inherent changes or deltas from baseline) in each parameter after 4 weeks or one year treatment with tiotropium or oxitropium were evaluated. Results Patients global impression (PI) after one year or 4 weeks were positively associated with physician global evaluation (PE) (r = 0.69 in both studies), with physician evaluation correlating more strongly with FEV 1 (r = -0.45 and -0.29) than patient impression (r = -0.27 and -0.06 respectively). For health status; however, associations of SGRQ with PI and PE were mores similar in magnitude (r = 0.38 and 0.35). Conclusion These data indicate that the direction and magnitude of associations of spirometry with health status in Japanese COPD patients is similar to those found in Western cohorts. The data also suggest that, in this clinical trial setting, overall health status as measured by SGRQ is better correlated with patient and physician assessments than FEV 1 . Breathlessness most often had the strongest correlations with primary outcomes, perhaps related to the chronic and troublesome nature of the sensation. Conclusion These data suggest that the smallest perceived change in patient status by overall patient assessment and physician global evaluation corresponds to a meaningful improvement in SGRQ scores, the magnitude of which supports an effect size of 4 units as clinically meaningful in a cohort of Japanese patient. Additionally, these data suggest that, in this clinical trial setting, the mean FEV 1 or FVC associated with the patient feeling better is less than the mean FEV 1 or FVC associated with the physician evaluation or patient impression. M ICHINOSE 1 , TJ WITEK 2 , M NISHIMURA 3 , A NAGAI 4 , K HIRATA 5 , M YOSHIDA 6 ,  Y FUKUCHI 7 Background COPD has been an increasingly recognized as a public health issue in Japan. However, the pattern of pharmacological therapy currently being utilized has not been well characterized.

Methods We examined data from two clinical trial cohorts at entry which evaluated the effect of the new inhaled COPD drug tiotropium. There were no medication requirements in the inclusion criteria with the only medication exclusion criteria being patients using oral steroids at a dose in excess of the equivalent of 10 mg prednisolone per day. Patients (n = 201 and 161) were on average (±SD) in the seventh decade (69.3 ± 6.9 and 68.7 ± 7.6) and had moderate to severe airflow obstruction (FEV 1 37.3 ± 13.7% and 36.8% ± 13.8% predicted).

Results Inhaled anticholinergics were the most frequently prescribed therapy with theophylline prescribed more frequently than beta agonists. Expectorant therapy was not uncommon, and inhaled steroids appeared to be used in a minority of patients.

The pattern of use as evident from a clinical trial cohort finds current therapy to be in line with current guidelines. Specifically, anticholinergics are used as first-line therapy and inhaled steroids are obviously used selectively relative to Western cohorts in the tiotropium program. Two exceptions to guidelines are the higher use of theophylline and expectorants. The pattern of therapy in patients treated by general practitioners needs to be evaluated. Results Significant improvement (P < 0.05) in performance was seen in 6MWD, symptoms of dyspnoea measured by Peak VAS and CRQ. The improvement was sustained at the time of follow up at 6 months. However, no additional significant improvement in performance was seen in the intervention group when compared to control. Conclusion Tiotropium therapy has improved health outcomes in patients presented with COPD in primary care settings. A 6 weekly Pulmonary Rehabilitation Programme did not give any additional benefits in patients already given tiotropium.

To assess neural respiratory drive in patients with chronic obstructive pulmonary disease (COPD) the oesophageal diaphragm EMG during CO 2 rebreathing and treadmill exercise was measured in seven COPD patients. The oesophageal catheter consisted of 9 consecutive recording electrode coils which formed 5 pairs of electrodes with an inter-electrode distance 3.2 cm within a recording pair. Each coil was 1 cm in length and the gap between adjacent coils was 0.5 mm. Maximal isometric contractions at functional residual capacity (FRC) and maximal voluntary inspirations from FRC to total lung capacity (TLC) were performed. All subjects underwent CO 2 rebreathing until end-tidal CO 2 was approximately 9% or they became intolerably breathless. Exhaustive treadmill exercise was performed at least 20 minutes after CO 2 rebreathing. There was a good linear relationship between root mean square of the diaphragm EMG (RMS) and end-tidal CO 2 (r = 0.92 ± 0.06) during CO 2 rebreathing. The amplitude of the RMS increased initially and then reached a plateau during exercise. We conclude that the diaphragm EMG is a useful technique to assess neural respiratory drive in patients with COPD and there may be an inhibition of respiratory centre output in patients with COPD during exhaustive exercise. Objective To study the change of diaphragmatic function of patients with COPD and the value of ultrasonography Evaluating diaphragmatic function.

Methods 34 subjects were divided into three groups: 14 patients diagnosed as COPD without history of respiratory failure were in emphysema group, 9 patients diagnosed as COPD with history of respiratory failure were in respiratory failure group, 11 subjects without history of respiratory and cardiac diseases were in control group. We measured the diaphragmatic thickness at the end of normal inspiration, normal expiration, forced inspiration and forced expiration, then calculated the average value. We also measured the diaphragmatic movement range during normal respiration and forced respiration.

The average diaphragmatic thickness were 5.6 ± 0.7 mm, 3.2 ± 0.5 mm and 4.5 ± 0.7 mm in emphysema group, respiratory failure group and control group respectively. The average diaphragmatic thickness in emphysema group was increased (P < 0.05) and that in respiratory failure group was decreased (P < 0.05). The diaphragmatic movement range during normal respiration were 18.7 ± 3.0 mm, 13.4 ± 3.1 mm and 17.1 ± 4.2 mm, and that during forced respiration were 32.2 ± 6.6 mm, 24.3 ± 5.7 mm and 30.9 ± 6.1 mm in emphysema group, respiratory failure group and control group respectively. The diaphragmatic movement range in respiratory failure group during normal respiration and forced respiration were all decreased (P < 0.05).

The diaphragmatic function in patients with COPD is normal or enhanced at early stage and is dropped at later stage. Ultrasonography is a useful non-invasive method for evaluating diaphragmatic function. Objective To investigate the association between genetic polymorphism of glutathione-s-transferases and susceptibility to chronic pulmonary disease (COPD). Methods The genotypes of GSTP1 were determined by Restricted FregmentLength Polymorphism (RFLP) and the genotypes of GST M1 and GST T1 were determined by multiplex polymerase chain reaction in patients with COPD and healthy smoking control subjects. Data were analyzed with SPSS 11.5. Results Three were no significant differences in the genetic polymorphism of GST P1, GST M1 and GST T1 between the two groups when ananlized separatly even adjusted by age, sex, and smoking index (P ജ 0.05). There was no significant differences in the genenotypes of combined GSTP1 A/A and GST M1 null, combined GST M1 null and GST T1 null genotypebetween the two groups even adjusted by subjects'age, sex and smoking index (P ജ 0.05). The proportion of combined GSTP1 A/A and GST T1 null genotype was significant higher in the cases than in the controls after adjusted by subjects'age, sex and smoking index (36.6% vs 26.6%, P = 0.03). Proportion of the genotype of combined GSTP1 A/A, GST M1 null and GST T1 null was significant higher in the cases than in the controls (24.1% vs 11.9%, P = 0.014). After adjusted by age, sex and smoking index, OR for this genotype versus all others genotypes was 2.74 (P = 0.007).

The results suggest that none of the GST genotypes had obvious effect on susceptibility to COPD when analysized respectively. However, there was probably synergetic effect between the genotypes of GSTP1 A/A and GST T1 null when each gene sites was combined to analysize. When all three gene sites was combined to analysize, there was probably synergetic effect among the three gene sites. Objective In China, there were different results for the levels of serum a 1 -AT reported by several investigators. No a 1 -AT genotype study has been reported. In this study, we will detect genotype and expression of a 1 -AT in the patients with COPD to investigate the effect of a 1 -AT on pathogenesis of COPD in Chinese.

Methods 132 COPD patients were studied. 110 subjects with normal pulmonary function were studied aas control. Genomic DNA was extracted from peripheral blood monocytes. Detection of PiZ allele in exon V and S allele in exon III were performed by PASA. The levels of serum a 1 -AT were measured by ELISA in 83 controls and 77 patients.

The mean age was 69.91 years old in COPD, and 66.97 years old in control group. The PCR production of S mutation was 285 base pairs. The PCR production of Z-mutation was 250 base pairs. The results showed that (1) All subjects were PiMM genotype of a 1 -AT, including COPD and control group.

(2) In control group, the mean serum a 1 -AT concentration was 2.88 ± 0.99 g/L in current smokers, and 2.60 ± 0.65 g/L in life time nonsmokers. The mean serum a 1 -AT concentration was 2.43 ± 0.84 g/L in patients with AECOPD and 2.23 ± 1.33 g/L in the COPD patients in the stable stage. Compared with control, the mean serum a 1 -AT concentration was significantly higher in AECOPD (P = 0.047), and in COPD patients in stable stage (P = 0.005) also. Conclusion There was no difference in genetic polymorphism of a 1 -AT between the patients with COPD and controls. However, the levels of serum a 1 -AT were significantly lower in the patients with COPD compared with control subjects. a 1 -AT deficiency may be a factor of the pathogenesis of COPD. The mechanism of a 1 -AT deficiency needs to be addressed.

Objective To explore the expression of NF-kB regulating MCP-1 in COPD rats and COPD patients, and investigate the effects of budesonide and theophylline on it.

Methods (1) The NF-kB activation in PBMC and the plasma concentrations of MCP-1 were measured by ELISA. The expression of NF-kB and MCP-1 in airway epithelial cells were detected by immunohistochemistry and in situ hybridization.

Results (1) The NF-kB activation in PBMC and the MCP-1 level of COPD rats were higher than that in normal rats (controls) (P < 0.01), and also higher than that in budesonide and theophylline group (P < 0.01).

(2) The percentages of positive cells for NF-kB as well as the MCP-1 mRNA and its protein expression in COPD rats were all higher than that in controls (all P < 0.01); (3) The percentages of positive cells for NF-kB in budesonide and in theophylline therapy group were lower than that in COPD rats (P < 0.01); The MCP-1 mRNA and the MCP-1 protein expression in budesonide and in theophylline therapy group were also lower than that in COPD rats (P < 0.01); (4) The NF-kB activation in PBMC and the plasma MCP-1 level in placebo group of COPD patients were higher than that in healthy group (P < 0.001), and also higher than that in budesonide and theophylline therapy group of COPD patients (P < 0.001).

The results suggested that NF-kB gene and its regulating protein MCP-1 may be involved in COPD. Glucocotiosteroid and theophylline may prolong the process of COPD. Objectives To explore the change of delayed rectifier potassium channel (K V ) activity in alveolar macrophages (AM) in chronic obstructive pulmonary disease (COPD) model rats for further elucidating the activation mechanisms of AM in COPD pathogenesis.

Methods COPD model was established by exposure to cigarette smoke. With whole-cell voltage-or current-clamp techniques, K V activity, membrane capacitance and resting membrane potential (Em) in AM from COPD model and control rats were compared. Results (1) Significant increases in total mononuclear cells and AM in bronchoal aveolar lavage fluid (BALF) were found in COPD group than those of control group;

(2) The AM K V current altitude in COPD group (520.5 ± 38.7 pA, +50 mV, n = 30) was significantly lower than that in control group (713.6 ± 44.4 pA, +50 mV, n = 30, P < 0.01); (3) AM from COPD group had no significantly different capacitances (P > 0.05), but had more positive Em (P < 0.01) compared with those from control group. Conclusion AM from COPD rats have inhibited K V function, more positive Em and higher excitability, which may be involved in the AM contribution to the pathogenesis of COPD. Background There is no an ideal method that can effectively prevent or delay the development of pulmonary emphysema. Retinoic acid (RA) plays an important role in the development and function of lungs. To study the effect and mechanism of RA on pulmonary emphysema, the models of experimental pulmonary emphysema in rats were made and intervenient effect of RA on them was observed.

Methods 21 Wistar rats were randomly divided into the normal control group (A), the model group (B), the RA treated group (C). Elastase was instilled into rat lungs of Group B, C, producing experimental emphysema. The rats in group C were treated daily by all-trons-retinoic acid. Pathological changes of the lung in rats were observed and the proliferating cell nuclear antigen (PCNA), the expression of Metalloproteinase-9 (MMP-9) and Tissue inhibitor of metalloproteinase-1 (TIMP-1) were examined. Results Compared with group A, mean alveoli area (MAA), mean lung interval (MLI), expression of MMP-9 and TIMP-1 in group B was significantly increased, but the ratio of MMP-9/TIMP-1 was unbalanced. Compared with group B, MAA, MLI, expression of MMP-9 and TIMP-1 in group C were significantly decreased, while the ratio of MMP-9/TIMP-1 was balance. The PCNA index staining in group C was increased than that in group B. Conclusion Early use of retinoic acid (RA) can alleviate the development of experimental pulmonary emphysema induced by elastase in rats. The intervenient effect of RA on the development of pulmonary emphysema may be related to its impact on the proteinase/antiproteinase ratio. 

The number of all cells, AMs and polymorphonuclear neutrophils (PMN) in BALF in patients with CB and COPD was significantly higher than that of the healthy subjects (P < 0.01, respectively). The concentrations of MMP-9 in BALF and the culture supernatants, the thickness of mucosa were significantly higher in COPD group than that in CB group, whereas they were apparently higher in CB group than that in the control group (P < 0.01, respectively). The MMP-9 activities in BALF of COPD group were positively correlated with those in the culture supernatants of their AMs (r = 0.529, P < 0.01); Both the concentrtion of MMP-9 in BALF and the positive number of cells expressing MMP-9 were positively correlated with the thickness of mucosa (r = 0.630, P < 0.05; r = 0.745, P < 0.01, respectively). Both the concentrtion of MMP-9 in BALF and the positive number of cells expressing MMP-9 were inversely correlated with forced expiratory volume in one second as percentage of predicted volume (FEV 1 %Pre) (r = -0.886, P < 0.01; r = -0.622, P < 0.05, respectively).

Conclusion AM MMP-9 may be involved in the occurrence and process of airway inflammation and remodeling. Objective To investigate the possibility of the trigger of the ventilator by EMG.

Method 4 subjects were involved in the study and the flow and the esophagus EMG were recorded in real-time during re-breath.

Results Both of the specificity and sensitivity of the trigger of ventilator by EMG were 100 percent. The opportunity of trigger by EMG was faster than the appear of flow and the difference between them was (20 ± 50) ms. Conclusion The EMG could be used to trigger the ventilator.

ZEGUANG ZHENG, RONGCHANG CHEN, NANSHAN ZHONG Guangzhou Institute of Respiratory Disease, Guangzhou, China 510120

Objective To investigate the effect of cardiac cycle on the esophageal pressure and the method of it's elimination.

Method 25 patients were involved in the study. The dynamical esophageal pressure and twitch esophageal pressure were measured in the sitting position, the supine positions before and after general anesthesia.

The changing value of dynamical esophageal pressure in the sitting position, the supine positions before and after general anesthesia was respectively (1.6 ± 1.1) cmH 2 O, (2.7 ± 1.2) cmH 2 O and (2.3 ± 1.2) cmH 2 O and there was a significant different between sitting position and the supine position before general anesthesia (P < 0.01), while there was not different between the supine positions before and after general anesthesia (P > 0.05); The twitch esophageal pressure during systole and diastolic phrase were respectively (16.3 ± 6.6) cmH 2 O and (14.1 ± 6.1) cmH 2 O and there was a significant different between them (P = 0.000); but after eliminated by mathematical filter, they were respectively (15.2 ± 6.2) cmH 2 O • (15.0 ± 6.0) cmH 2 O and there was not different between them (P > 0.05). Conclusion The esophageal pressure was effected by cardiac cycle and the effect was larger in lying position than in sitting position. The negative pressure of thoracic cavity did not affected the effect of cardiac cycle on the esophageal pressure. The twitch esophageal pressure overestimated the function of diaphragm during systole phrase and underestimated it during diastolic phrase and the effect of cardiac cycle on the esophageal pressure could be eliminated by mathematical filter. Results In-hospital mortality was 52.5%. Single factor analysis indicated that age, total hospital time, intensive care time, actived partial thromboplastin time, red cell count, blood glucose level, presence of gastrointestinal hemorrhage, Acute Physiology and Chronic Health Evaluation (APACHE II) scores and undergoing tracheotomy were risk factors relevant to in-hospital death (P < 0.05). By means of multiple logistic regression, Independent predictors of death were: higher age (P = 0.049; OR 1.032; 95% confidence interval: 1.00~1.065), higher level of blood glucose and APACHE scores (P = 0.005; OR 1.124; 95% confidence interval: 1.037~1.219, P = 0.034; OR 1.085; 95% confidence interval: 1.006~1.169), the total hospital time (P = 0.009; OR 0.959; 95% confidence interval: 0.930~0.989) was independent predictors of benignantly outcome. Conclusion Survival after mechanical ventilation is poor and can be estimated by using clinical variables including age, total hospital time, level of blood glucose and APACHE scores.

Background Respiratory muscles weakness can cause respiratory failure and contribute to weaning difficult in intensive care unit. Measurement of diaphragm compound muscle action potential (CMAP) provides useful information on diaphragm function. Conventional methodology for measurement of the diaphragm CMAP is electrical stimulation and chest wall surface electrode, which is potentially difficult to widely use in intensive care unit (ICU) because electrical stimulation of the phrenic nerve is technically demanding and the surface electrode can be unreliable in recording the diaphragm CMAP. Methods 10 normal subjects and 10 patients in ICU were studied. The diaphragm CMAP was measured with a multipair esophageal electrode during electrical and magnetic stimulation of the phrenic nerve. Results Good quality CMAP was recorded from both normal subjects and patients in ICU. In normal subjects, the phrenic nerve conduction time (PNCT) measured with electrical stimulation was similar to that measured with magnetic stimulation (7.4 ± 0.9 ms vs 7.5 ± 0.8 ms, P > 0.05 on the left side; 7.0 ± 0.8 ms vs 6.9 ± 0.8 ms, P > 0.05 on the right). The amplitude of the CMAP measured with electrical stimulation was also similar to that measured with magnetic stimulation (1.54 ± 0.43 mV vs 1.61 ± 0.41 mV, P > 0.05 on the left side; 1.49 ± 0.39 mV vs 1.50 ± 0.36 mV, P > 0.05 on the right). No CMAP response to magnetic stimulation was found on one side or both sides in three patients who had difficulty in weaning from mechanical ventilation. The amplitude of the CMAP in the patients who were able to respond to magnetic stimulation was smaller than that in the normal subjects (0.71 ± 0.36 mV vs 1.58 ± 0.38 mV, P < 0.001, pooled left and right values). Only one of the ten ICU patients had a prolonged PNCT. Conclusion This study suggests that magnetic stimulation is a useful technique in assessment of diaphragm function in ICU. Unilateral or bilateral diaphram paralysis contributes to difficulty in weaning from mechanical ventilation. We hypothesized that the vascular pedicle width on supine, portable chest X rays could be used to predict intravascular volume overloaded status in critically ill patients. We used pulmonary artery occlusion pressure measurements ≥18 mmHg as indicative of a fluid overloaded state and measurements <18 mmHg as normal/low volume states. Recognizing the limitations of the PAOP, this is still the standard invasive method of determining volume status in the intensive care unit and so was chosen as the reference standard. Traditionally invasive haemodynamic measurements of pulmonary artery occlusion pressures have been used to assess the volume status in critically ill patients. The vascular pedicle, as seen on chest X ray is the mediastinal silhouette of the great vessels. This can be measured by dropping a perpendicular line from the point at which the superior vena cava crosses the right mainstem bronchus across to the point where the left subclavian artery exits the aortic arch, This measurement is called the Vascular pedicle width, with a reported mean 'normal' value of 48 ± 5 mm in healthy upright volunteers. Milne E, Pistolesi M et al. Radiology 1984;152:1-8. We hypothesized that the vascular pedicle width (VPW) on supine, portable chest X rays could be used to predict intravascular volume overloaded status in critically ill patients. We conducted a prospective, blinded trial where both pulmonary artery occlusion pressures and vascular pedicle widths were measured in patients admitted to the intensive care unit. We used pulmonary artery occlusion pressure (PAOP) measurements ≥18 mmHg as indicative of a fluid overloaded state and measurements <18 mmHg as normal/low volume states. Standardized, portable chest X rays in the supine position were obtained within an hour of PAOP measurement. Receiver operating characteristics curves were constructed using different cutoffs of the vascular pedicle width measurement to identify sensitivities and specificities for each value. Measurements were obtained from forty-five patients. Using ROC derived cutoffs, a vascular pedicle width measurement of 72 mm was found to have a sensitivity of 67% and a specificity of 58% for correctly predicting a fluid overloaded state. These data suggest that the known correlations of VPW with intravascular volume in upright X rays may be extended to the portable X rays available in the intensive care unit.

Respiratory muscles weakness is cause of respiratory failure and contributes to weaning difficult in intensive care unit. measurement of diaphragm Compound muscle action potential (CMAP) provides useful information on diaphragm function and the management of mechanical ventilated patients. Conventional methodology for measurement of the diaphragm CMAP is electrical stimulation and chest wall surface electrode, which are potentially difficult to wildly use in Intensive Care Unit because electric stimulation of the phrenic nerve is technique demanding and the CMAP recorded from surface electrode can be contaminated by other muscle activity. To investigate whether unilateral magnetic stimulation and multipair esophageal electrode can be used to measure diaphragm CMAP in ICU, 10 normal subjects and 10 patients in ICU were studied, we measured the diaphragm CMAP during electric and magnetic stimulation of the phrenic nerves using multipair esophageal electrode. Good quality CMAP was measured from both normal subjects and patients in ICU. In normal subjects, The phrenic nerve conduction time (PNCT) measured with electric stimulation was similar to that measured with magnetic stimulation (7.4 ± 0.9 ms vs 7.5 ± 0.8 ms, P > 0.05 on the left side; 7.0 ± 0.8 ms vs 6.9 ± 0.8 ms, P > 0.05 on the right). The amplitude of the CMAP measured with electric stimulation was also similar to that measured with magnetic stimulation (1.54 ± 0.43 mV vs 1.61 ± 0.41 mV, P > 0.05 on the left side; 1.49 ± 0.39 mV vs 1.50 ± 0.36 mV, P > 0.05 on the right). No CMAP response to magnetic stimulation was found on one side or both sides in three patients who had difficulty in weaning from mechanical ventilation. The amplitude of the CMAP in the ICU patients who were able to respond to magnetic stimulation was smaller than that in the normal subjects (0.71 ± 0.36 mV vs 1.58 ± 0.38 mV, P < 0.001, pooled left and right values). Only one of the ten ICU patients had a prolonged PNCT. This study suggested that magnetic stimulation is a useful technique in assessment of diaphragm function in ITU. Unilateral or bilateral diaphram paralysis contributed to difficulty in weaning from mechanical ventilation. Objective To evaluate the severity of airflow obstruction in patients supported with mechanical ventilation and the effect on respiratory mechanics after salbutamol inhalation by analysis parameters of tidal breathing flow-volume curves (TBFV). Methods: 26 patients (age 67 ± 5), intubated, sedated and mechanically ventilated at least 24 hour for acute respiratory failure of diverse causes (chronic obstructive pulmonary disease, COPD and asthma). After 20 mins with volume controlled ventilation (SIMV mode, tidal volume 7~9 ml/kg), Measurements of TBFV loops and respiratory mechanics [including compliance (Cst), airway resistance (Raw) and instrinic PEEP, (PEEPi)] were operated and repeated again after salbutamol 600 mg inhalation. Results: After salbutamol inhalation, all patients Raw was decreased significantly (21.9 ± 2.4 cmH 2 O/L·s versus 13.1 ± 1.8 cm H 2 O/L·s, P < 0.05), also PEEPi was lower from 6.9 ± 1.2 cmH 2 O to 4.5 ± 1.1 cmH 2 O(P < 0.05). peak tidal expiratory flow (PTEF) and the fraction of exhaled volume to achieve PTEF to V TE (V PTEF /V TE ) were increased slightly, but the fraction of exhaled time to achieve PTEF to T E (T PTEF /T E ), tidal expiratory flow at 50% of the remaining tidal volume/PTEF(TEF50/PTEF) and tidal expiratory flow at 25% of the remaining tidal volume/PTEF (TEF25/PTEF) were significant increased (P < 0.05). TEF50/PTEF, TEF25/PTEF had a significant correlation with PEEPi (r = 0.59 and 0.54, P < 0.05) than other TBFV parameters. The correlations were diminished lightly after salbutamol inhalation. Conclusions: The measurement of TBFV loops was simple and need not special technique, The degree of airflow obstruction and the effect of bronchodilator drug in mechanical ventilated patients could be known by analysis of TBFV parameters. Inhaled nitric oxide (iNO) is used to treat hypoxemia associated with acute lung injury. We hypothesized that iNO may alter alveolar liquid clearance (ALC) in ventilated rabbits with lung injury induced by endotoxin. Eighteen rabbits were injected with endotoxin and ventilated 4 hours later. Volume control ventilation (Vt 15 ml/kg, f 40 bpm, PEEP 5 cm H 2 O) were given and all animals were randomly assigned into three groups: ventilation only, ventilation with iNO 40 ppm and ventilation with iNO 10 ppm. Three hours later isotonic albumin labeled by 125 I was injected into the right lower lobe and albumin labeled by 131 I into blood. 131 I radioactivity in plasma was detected as the baseline after ten minutes blood mixturing. One hour after injecting radionuclide, the radioactivity of 125 I and 131 I in alveolar liquid from right lower lobe and plasma were detected. Results were shown in table (W/D ratio means wet-to-dry lung weight ratio). We conclude that iNO can reduce lung edema by increasing ALC, and the low concentration NO inhalation may be more effective than high concentration in treatment of lung injury. Objective To determine whether hypercapnia is protective against oleic acid-induced acute lung injury (ALI) model. Moreover, to examine the effect of hypercapnia on NF-kB and TNF-a. Methods Twenty-two New Zealand rabbits were randomly allocated to control group (CON), normocapnic group (NC) and hypercapnic group (HP). The latter two groups were injected with oleic acid (0.1 ml/kg) intravenously to establish ALI model. Then TNF-a concentration in serum and bronchoalveolar lavage fluid (BALF) and the expression of NF-kB in the lung tissue were analyzed. Lung mechanics, hemodynamics, blood-gas analysis indices and lung wet : dry ratio, BALF protein were measured. Histologic damage was assessed after 3 hours' mechanical ventilation.

Results TNF-a concentration in serum and BALF in HP group was lower than that in NC group (P < 0.05). The expression of NF-kB in HP group was less than that in NC group by both immunohistochemistry and Western-blot analysis. Peak airway pressure, lung dynamic compliance and PaO 2 in HP group were significantly improved compared with NC group (P < 0.05). Lung wet : dry ratio and BALF protein were significantly lower in HP group than that in NC group (P < 0.05). Histologic damage in NC group was significantly severer than that in HP group. NF-kB was correlated with PaCO 2 , TNF-a concentration in serum and BALF (r = -0.568, 0.516, 0.520, P < 0.05, respectively).

Conclusions Hypercapnia decreased the severity of lung injury in this in vivo ALI model within the first 3 hours after oleic acid administration. The mechanism might be associated with the inhibition of NF-kB activation and then the decreased expression of TNF-a. 

Object To know about the clinical and radiological manifestation of pulmonary alveolar proteinosis and improve the diagnosis ability of this rare disease through clinical cases analysis. Method Analysis 10 patients with confirmed PAP involved from 1991 and their clinical data retrospectively. Result (1) there were 8 males and 2 females with the average age of 49.7 years old and most of them were from 40 to 50 years old. (2) PAP presented a protracted chronic clinical course but few physical signs. Their major complains included dyspnea on exertion, cough. (3) Some of them were exposed to metal dusts and special chemical substance. (4) Chest plain radiography showed diffuse airspace opacities, most located in the lower lobes, similar to cardiogenic pulmonary edema. Characteristic chest CT showed 'geographic' or 'crazy paving' changes. (5) cases were diagnosed by transbronchoscopic lung biopsy (TBLB) and brochoalveolar lavage fluid (BALF), 1 by BALF with periodic acid-schiff staining and electromicroscopic examination, 4 by thoracscopy lung biopsy. Conclusion It should be alert to PAP when the following happen: bilateral diffuse airspace opacities, most located in the lower lobes, and is similar to cardiogenic pulmonary edema in chest X-ray; 'geographic' or 'crazy paving' changes in chest CT; middle age patient, especially male; the longer duration of disease, dyspnea on exertion with absence of obvious lung physical sign. And patient should undertake the examination of TBLB, BALF, thoracscopy lung biopsy if necessary at the beginning to enhance the diagnostic accuracy ratio. Background The clinical course of idiopathic pulmonary fibrosis (IPF) is usually chronic and slowly progressive with a median survival of 3-5 years. However, some patients experience rapid deterioration during their clinical course. This phenomenon, acute exacerbations is poorly described and variably defined. The aim of this study is to assess the course of this phenomenon and to estimate its relative frequency.

The clinical, imaging and pathologic data from 16 of 147 patients with IPF-UIP who suffered acute exacerbations were reviewed. Result The mean age was 63.1 ± 9.6 years with a male predominance (12 : 4). Duration of IPF before acute exacerbations was 20.2 ± 21.9 months. Time from onset of acute symptoms to admission was 20.8 ± 17.7 days. Eleven of the 16 patients had more severe disease clinically satisfying the (stricter) criteria of Kondoh et al. although all biopsies taken during acute exacerbations showed DAD superimposed upon UIP, regardless of the criteria used. There was no significant clinical difference between patients defined by either set of criteria, except duration of acute symptoms and initial PaO 2 /FiO 2 ratio. Imaging showed diffuse bilateral ground glass opacities superimposed on subpleural reticular and honeycombing densities. Peripheral distribution of opacities and higher PaO 2 /FiO 2 ratio appeared to correlate with a better survival. Conclusion Acute exacerbations of IPF are more frequent than previously thought and the process encompasses a spectrum of severity rather than being a single defined phenomenon.

Objective The aim of this study was to investigate the role of cells in bronchoalveolar lavage fluid (BALF) samples on pulmonary function in patients with idiopathic pulmonary fibrosis (IPF). Methods Thirty two patients with IPF underwent pulmonary function tests (PFTs) and fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) at rest. HE stain was used for cell populations analysis. Results There was a significant negative correlation between the percentage of eosinophils (Eos%) in BALF samples and %VC (r = -0.37, P < 0.05) or %D L CO (r = -0.47, P < 0.01), no such relationship was demonstrated for the percentage of any other cell type. An inverse relationship was found between V 50% or V 25% from pulmonary function and the percentage of neutrophils (Neu%) (r = -0.66, P < 0.01; r = -0.45, P < 0.05, respectively) in BALF samples. Conclusion The results suggest the possibility that eosinophilic cells and neutrophilic cells may contribut to pulmonary dysfunction in patients with IPF.

Background There is a trend that the incidence of diffuse lung diseases (DLD) has been increasing in recent years. But China lacks a comprehensive survey until now. We analyze 395 subjects with DLD administrated in our hospital from 1990 to 2003 and hope it can contribute to a large-scale survey of DLD in China. On the other hand, we hope it can help Chinese doctors diagnosing these kinds of disease more correctly. Methods 395 patients (190 of male and 205 of female, mean age 56.09 ± 12.98 years) who with diffused pathology on their conventional chest radiography and computed tomography (CT) scan and administrated in our hospital from 1990 to 2003 were all enrolled in the study. Their age, sex, professional, smoking or not, diagnosis, biopsy data, the number of hospitals that they ever administrated, how long they spend from initial to final diagnosis, the effect of therapy and other characteristics were analyzed. Results 1. The number of DLD patients administrated to our hospital and biopsy rate increased very evidently in recent 6 years, respectively 337 of 395 administrated cases and 92 of 102 biopsy cases. 2. The ratio of male to female was 0.93, but it varied according to different diseases, some kinds of disease have higher ratio, such as usual interstitial pneumonia (UIP) 1.87, diffuse panbronchiolitis (DPB) 1.5, respiratory bronchiolitis-associated interstitial lung disease (RBILD) 1.33; other diseases have lower ratio, such as cryptogenic organizing pneumonia (COP) 0.25, nonspecific interstitial pneumonia (NSIP) 0.29, sarcoidosis 0.36, connective tissue diseases 0.24. 3. The number of our patients increased with older age, such as 143 of 395 was over 60 yr (36.2%), 113 of them were 51 to 60 yr (28.6%), 80 of them were 41-50 yr (20.3%). 36 of 43 cases with UIP were over 50 yr (83.7%), 12 of 22 cases with NSIP were 41-50 yr (54.5%). 4. 142 subjects had some kinds of specific profession, such as 60 of 142 (42.3%) were peasants, 22 of 142 (15.49%) worked in textile mill. 5. 137 (34.7%) subjects were smoker and the diseases with higher smoking rate included lung cancer (72.7%), DPB (60%), RBILD (57.1%) and UIP (48.8%). 6. The effect of therapy were different with the kinds of DLD, 102 patients had surgical lung biopsy and their effect rate of therapy was 76.47% (78/102), but it varied with different diseases, some diseases had 100% of effect, included DIP, BOOP, RBILD, DPB and sarcoidosis, but NSIP was 84.2%, UIP just was 40%. 7. There were big differences among the time span from initial stage to final diagnosis, some less than one month, some over thirty years, middle time was thirteen months. The number of hospitals that they ever administrated varied from one to eight, average 2.7. Conclusion Interstitial lung diseases (ILD) expect UIP have better response to steroid therapy. Doctors have to understand ILD more clearly, diagnose more correctly and earlier. Idiopathic nonspecific interstitial pneumonia (NSIP) is a distinct disease entity showing much better treatment response and long-term prognosis than usual interstitial pneumonia. Unfortunately little is known about the pattern of short-term clinical improvement and relapse of NSIP after completion of corticosteroid and/or cytotoxic therapy. Total 42 patients (mean age; 53.4 ± 9.5 years, M : F = 7 : 35) with idiopathic NSIP, proved by surgical lung biopsy between July 1997 and July 2003, were retrospectively reviewed. Treatment responses after starting treatment were assessed at 3rd month and 6th month. If the patients met any two of following three criteria, they were considered to show a favorable response:

(1) reduction of dyspnea grade and/or cough (2) improvement of chest PA/HRCT findings (3) increase of FVC or TLC > 10% or DLco > 15%. Relapse of lung lesion was determined by symptom change, radiographic findings and pulmonary function tests. At 3rd month after treatment, 30 patients (71.4%) showed a favorable response and the other were stable. Improvement of FVC (13.1% predicted value) was remarkable as well as reduction of dyspnea. Thirty-six patients (85.7%) showed a favorable response at 6th months, with marked increase of FVC (16.9% predicted value) and decrease of ground-glass opacity (11.9 %) on chest HRCT. Among the patients (n = 21) who were followed (mean; 38.4 months) longer than one year after stopping corticosteroid and/or cytotoxic therapy, 12 patients (57.1%) showed relapse of lung lesion. Mean interval of relapse after stopping medication was 15.1 months (2~40 months). There was no significant difference in pre-treatment factors between relapsed group and non-relapsed group.

In conclusion, idiopathic NSIP, although showing good treatment response, recurs in more than half of patients after discontinuation of medical treatment. Background Sarcoidosis is a multisystem granulomatous disorder of unknown origin. Since young adults suffer from sarcoidosis more frequently than the elderly, the clinical feature of patients who diagnosed at older ages has not been fully established. In order to characterize elderly patients with sarcoidosis, the clinical, radiographic and laboratory features of patients with sarcoidosis diagnosed at 65 years of age or older were compared to those of patients diagnosed at younger ages. Methods The medical records of 401 patients with sarcoidosis referred to our department between January 1985 and December 2003 were reviewed. They were divided into 3 groups; group A (patients diagnosed at 65 years of age or older, n = 59), group B (patients diagnosed between 40 and 65 years of age, n = 140), and group C (patients diagnosed under 40 years of age, n = 202). The clinical feature of group A patients was compared with that of group B and C patients.

The female to male ratio of group A patients was significantly higher than that of group C patients. Eye and skin involvements were seen more commonly in group A than in other groups. In contrast, the incidence of serious extra-pulmonary involvements (e.g., cardiac, central nervous system diseases etc.) in group A patients was lower than that of other groups. There was no difference among three groups with regard to laboratory findings (e.g., serum angiotensin-converting enzyme level, serum levels of gamma globulin, and calcium). The percentage of group A patients, who were treated with oral corticosteroids, was lower than that of other groups patients.

Conclusion This study suggests that the clinical feature of sarcoidosis in elderly patients differ from those seen in younger patients.

Background Our previous study has indicated that okam extract J201 exerts inhibition on the proliferation of Human Lung Fibroblast. Its efficacy on suppression of pulmonary fibrosis in intact animal and underlying mechanism of action are further investigated. Methods Animal model of pulmonary fibrosis is induced by intratracheal administration of bleomycin. Okam is administered at dose ranging from 25, 50, 100 mg/kg. Lung pathology and MDA and hydroxylproline content in the supernatant of lung tissue were evaluated at end of the study. In vitro activity of okam extract J201 on macrophage suppression was evaluated and magnetic resonance was applied to investigate its interaction with PDGF.

Results Okam extract J201 at dose of 50 mg/kg significantly inhibits lung inflammation and pulmonary fibrosis. The lung fibrosis indexes in animal model and okam groups were 1.23 ± 0.13 and 0.99 ± 0.11 respectively (P < 0.05). MDA and hydroxylproline content in the supernatant of lung tissue in okam group were significantly lower than those in animal model group, being 6.50 ± 1.26, 2.35 ± 0.58 and 5.07 ± 1.01, 1.78 ± 0.30 respectively (P < 0.05). TNF-a secretion by macrophage in BALF was suppressed by okam J201. Magnetic resonance showed interaction between okam J201 and PDGF-BB with binding affinity coefficient K D of 2.94E-11. Conclusion Our data show that okam extract J201 inhibits bleomycin- Background Bunashimeji mushroom is cultivated in an indoor environment all year round. Employees who worked here have often suffered from cough, sputum and shortness of breath. This study aimed to clarify the clinical features and therapy of this disease. Methods Fifteen patients had mild respiratory symptoms of dry cough, sputum, and low-grade fever. We diagnosed them as HP from clinical features, HRCT findings, BALF findings, lung histology, and lymphocyte stimulation test (LST) for Bunashimeji. Five of 15 patients ('avoidance') quit their job to avoid exposure to spores after hospitalization. As the therapies, Fifteen patients were divided into two subgroups; 'mask alone' (3 patients) and treatment with prednisolone orally with mask ('mask+PSL', 7 patients) for 2 months. Results PFTs revealed a slight decrease in the percentage diffusing capacity for carbon monoxide. Four of 15 patients showed normal findings on the chest X-ray, but HRCT findings of all patients showed diffuse groundglass opacities with centrilobular nodules. BALF revealed an increase in total cell counts, showing predominantly activated lymphocytes. The results of the LST were positive in all cases. The 'avoidance resulted in a significant decrease in LST (P < 0.05). In all groups, chest HRCT showed partial disappearance of the ground glass opacity and the presence of small nodules in the short-term. Total cell counts in BALF of the 'mask+PSL' group significantly decreased after treatment compared with the 'avoidance' and 'mask alone' groups (P < 0.05 and P < 0.01, respectively).

Conclusions Chest HRCT and BALF showed the typical subacute phase of HP although patients with HP have mild respiratory symptoms. Treatment with prednisolone gave a more rapid improvement in these patients as assessed by the high serum KL-6 and SP-D concentrations, and severe ground glass opacity.

In sarcoidosis, an inflammatory lung disease, the CD4/CD8 ratio of bronchoalveolar lavage fluid (BALF) is altered. To compare BALF CD4/CD8 in sarcoidosis of stage I and stage II, samples from six patients of stage I and eleven of stage II were analysed. A comparison of the CD4/CD8 showed no significantly deference between sarcoidosis of stage I and stage II (2.65 versus 2.58), although the ratioes are higher than normal value (1.45 ± 0.79). In conclusion, the CD4/CD8 ratioes in BALF is similar in sarcoidosis patients of stage I and stage II. We suggest there is similar inflammatory status in alveolars of sarcoidosis patients of stage I and stage II. Background The long-term prognosis of hypersensitivity pneumonitis is controversial. In order to elucidate the long-term prognosis of farmer's lung, as the representative of occupational hypersensitivity pneumonitis, we conducted a follow-up survey of cases with acute farmer's lung.

The subjects were 6 cases who developed acute farmer's lung in the period from 1982 to 1991. These cases corresponded to the Standard of Diagnosis of hypersensitivity pneumonitis compiled by the Ministry of Health and Welfare of Japan based on environmental factors, positive serum precipitin against Saccharopolyspora rectivulgura or Thermoactinomyces vulgaris and pathological findings of lung biopsy. The patients' age was 56.2 ± 6.79 years (44 to 63) at acute onset. All patients showed improved Symptoms, chest X-ray and CT findings and attained remission through hospitalized treatment. After 10 to 20 years (14.8 ± 4.90), we conducted follow up survey. All patients were still engaged in dairy farming. All subjects never smoked. Results All of the 6 cases showed abnormal chest CT such as small bullae, fine nodules, fine granular shadows, ground-glass opacity or emphysematous changes. One showed honeycombing. Conclusion Even if acute symptoms of farmer's lung improve, chronic exposure to antigens has the risk to induce fibrosis and emphysematous changes of lungs. Bronchiolitis Obliterans (BO), a clinico-pathologic diagnosis of bronchiolar inflammation and fibrosis with pregressive dyspnea, occurs commonly in patients with rheumatoid arthritis (RA) de novo or as a complication of Dpenicillamine treatment. Bucillamine, another disease modifying antirheumatic drug similar to D-penicillamine in molecular structure in that both contain a sulfhydryl group, has been reported to be associated with lung injury. None of these lung injuries were BO to our knowledge. 57 year-old female was diagnosed of seronegative RA, and was treated with 200 mg of bucillamine daily from December 2000. Three months later, she presented complaining of cough, mucoid sputum and exertional dyspnea. Chest x-ray revealed reticular opacities on both lung fields. HRCT showed peribronchial fibrosis with bronchial wall thickening, bronchiectasis, bronchioloectasis and some centrilobular nodules with branching linear pattern in both lung fields. Lung biopsy by video-assisted thoracic surgery showed marked thickening of peribronchioles with extensive proliferation of smooth muscle and narrowing of bronchiolar lumina, which are consistent with constrictive bronchiolitis. Bucillamine was discontinued, and the patient improved symptomatically and radiologically on serial chest X rays. We believe that this is the first report of bucillamine-induced bronchiolitis obliterans pathologically proven. Larger scale study will offer more insight to this condition.

Methods Wistar rats were instilled bleomycin (5 mg/kg) via trachea. The animals were killed on 1, 3, 7, 14 and 28 days after the administration of bleomycin or saline. Immunohistochemistry assay for MMP-2 (gelatinase A), MMP-9 (gelatinase B) and TIMP-1 were performed in lung tissue.

Result (1) MMP-2 expression preferentially in the alveolar epithelial cells and the bronchial epithelial cells (2) MMP-9 expression in the alveolar macrophages, interstital macrophages, neutrophils, bronchiolar epithelial cells.

(3) The expression of MMP-2 and MMP-9 was stronger on day 3 to day 7 compared with that on remaining days and the controls. Objective To evaluate HRCT diagnostic value for nonspecific interstitial pneumonia (NSIP) and to correlate CT findings with pathology.

Methods HRCT manifestation of 20 cases of NSIP were evaluated by two independent observers included the presence, extent, and distribution of various HRCT findings. ALL cases were confirmed by operation and pathology. The comparsion of HRCT findings and pathology was done.

The predominant findings observed on initial HRCT in all patients were patchy areas of ground-glass attenuation present with areas of consolidation, which present bilaterally in the middle and lower lung zones. It can be seen in all patients. Interlobular reticular opacities and traction bronchiectasis were seen in 16 and 12 patients, respectively. The pathological characteristic was uniform interstitial inflammation and fibrosis. 16 had cellular and fibrosing patterns; 4 had fibrosing patterns only; the former showed better response to glucocortcoid therapy than the latter. Conclusion HRCT findings of NSIP are well corresponded with pathology. HRCT examination can help to make the diagnosis and differential diagnosis. Definite diagnosis depends mainly on open lung biopsies. HRCT is very useful for choosing the perfect treatment ways. Rationale 14-membered ring macrolides (14-MRMLs) have been reported to improve the survival of patients with DPB by anti-inflammatory mechanisms. We previously reported that 14-MRMLs play to preventing lung injury and fibrosis in bleomycin-challenged mice. EM703 is new derivative of erythromycin. It is twelve-membered ring macrolides, and hasn't antibacterial action. The present study was undertaken to investigate the effects of EM703 on an experimental model of bleomycin-induced acute lung injury and subsequent fibrosis in mice.

Methods Bleomycin was administered IV to ICR mice. EM703 was orally administered daily. At 28 days after bleomycin injection, fibrotic foci were histologically observed, and hydroxyproline (HOP) content was chemically analyzed in lung tissues. Cell populations in BAL fluid were examined at 7 days after BLM injection. Expressions of messenger RNA (mRNA) of TGFbeta and Smad3 in lung tissues were examined at 7, 14, 21, 28 days after bleomycin treatment. The inhibitory effect of EM703 was assessed by overall comparison between control' bleomycin alone, and bleomycin plus EM703 groups. We further examined whether the effect of EM703 for the proliferation of mouse lung fibroblasts with Cell counting Kit-8 in an in vitro cell line MLg 2908 culture system. Results Bleomycin-induced pulmonary fibrosis was inhibited by EM703 on day 28 after bleomycin injection. Macrophages and neutrophils infiltration into the airspace were inhibited by EM703 on day 7 after bleomycin injection. Expression of Smad3 mRNA tended to be attenuated by bleomycin, but recovered by together with EM703. EM703 inhibited proliferation of MLg 2908 induced by TGF-beta in vitro.

Conclusions These findings suggest that EM703 may be modification of intracellular TGF-beta signaling by Regulation of Smad3 mRNA. EM703 may directly inhibit proliferation of lung fibroblast induced by TGF-beta and development of fibrosis. This may be one mechanism of the antifibrotic effects of EM703. Objective To evaluate the role of IL-13 in peripheral blood and BALF from patients with idiopathic pulmonary fibrosis (IPF). Methods With ELISA, the level of interleukin 13 (IL-13) was determined in the serum and BALF of 17 IPF and 8 non-interstitial lung disease subjects (nonsmokers).

The level of IL-13 of BALF in the patients with IPF was significantly higher than that in non-interstitial lung disease group [(301 ± 86) ng/L vs. (103 ± 24) ng/L, P < 0.01] and was higher than that in serum [(178 ± 36) ng/L vs. (55 ± 15) ng/L, P < 0.01]. The level of IL-13 in the BALF of patients with IPF was positively correlated with the percentage of neutrophils (r = 0.84, P < 0.01). The analysis of spearman correlation showed that the level of BALF IL-13 was correlated with lung function and PaO 2 , D L CO (spearman correlation coeffieient r = -0.898, -0.878, -0.874, -0.890, respectively, P < 0.01).

Conclusions IL-13 might play an important role in the pathogenesis of IPF and act as the potential marker of activity.

Background Accidental foreign body inhalation is not uncommon. We report a series of cases highlighting complications, and difficulties encountered at retrieval. Methods Retrospective review of inhaled foreign body (IFB) cases from a single institution over 12 (1991) (1992) (1993) (1994) (1995) (1996) (1997) (1998) (1999) (2000) (2001) (2002) (2003) years. Results Eight cases of IFB were identified (table) . Elderly cases had co-morbidities and inhaled food whereas young patients inhaled inanimate objects. Fibreoptic bronchoscopy was successful in 7 cases. Two cases had laryngeal mask airway for ventilation during bronchoscopy. Difficulties at removal were frequently encountered. Objective To value the outcome of the protective sheath of fiberoptic bronchoscope (applied patent number: 011069600). Via it diagnosing bacterial organisms of pulmonary infections. Methods The protective sheath of fiberoptic bronchoscope was made of polythylene including three parts: joints, Channel and cap that can be opened by fiberoptic bronchoscope after passing the rimalottidis. Fiberoptic bronchoscope examinations with the protective sheath were performed in 25 patients with pulmonary infections and 20 patients without it. Specimens collected by protected specimens brush (PSB), Single Sheathed catheter brush (SSC) and bronch-alveolar lavage (BAL) were quantitatively cultured while expectorated sputum was qualitively cultured. Tests of drug susceptivity were performed too.

Results 45 fiberoptic bronchoscopic examinations were successfully performed, all patients can endure it without discomfort. with the cut off of 10 3 colonary forming unit (cfu)/ml, 10 3 cfu/ml and 10 4 cfu/ml, The sensitivity of PSB, SSC and BAL are (14/25) 56% (CI 35~36%), (16/25) 64% (CI 42~82%) and (20/25) 80% (CI 59%~93%) respectively, BAL via the protective sheath is better than that of PSB, P < 0.001, t = 4.956, the specificity of PSB, SSC and BAL is 100%, 95% and 95% respectively, there is no significant difference between them. The joint of SSC and BAL via the protective sheath of fiberoptic bronchoscopy are better than that of PSB, P < 0.005, x 2 = 9.33. Conclusions the protective sheath of fiberoptic bronchoscopy is of good diagnostic value in SSC and BAL in pulmonary infections, it can be easily, convenientily, safely and cost-effectively applied with promising future. Objective To detect and analyze the relationship between the improvement rate of FVC, TLC, FEV1, DLco and the percentage of ground glass opacity and honeycombing shadow in HRCT, and evaluate the role of quantitative analysis of HRCT in diagnosis of Idiopathic Interstitial Pneumonia (IIP).

Methods In-patients with IIP were collected from 1997 to 2003 in our hospital. From these patients, 21 cases, who took steroid treatment and had integrated materials, were used to make retrospective analysis. The diagnosis was based on pathologic data if the patient had lung biopsy. Clinical diagnosis was made according to clinical criteria, and the other diseases had been excluded. Initial HRCT scans obtained at the time of diagnosis was used, before initiation of corticosteroid treatment, to determine the HRCT score. Four sections of HRCT scans were analyzed, including the level of the aortic arch, tracheal carina, left atrium, and diaphragmatic dome, by radiologists who were blinded with respect to the clinical and pathologic data. HRCT scans were dealt with Adobe Photoshop software to distinguish abnormal parts from the normal ones and programmed using Visual C++ to read image pixels. The percentage of lung parenchyma change that showed ground-glass attenuation and honeycombing shadow was calculated on each of the four high-resolution sections. The overall percentage was calculated by averaging the percentage for each section. HRCT scores were compared with the improvement rate in FVC, TLC, FEV1 and DLco by using the Spearman rank correlation coefficient in SPSS 11.5 software. Sequential variable was tested by t-test.

There was only 27.1% of patients underwent lung biopsy for pathologic diagnosis in all 107 cases from 1997 to 2003. 31 cases were treated with corticosteroid. 21 cases were investigated, and divided into two groups, including group A (corticosteroid-responsive patients) and group B (no steroid-responsive). (1)

The mean age was 59.1 ± 13.0 years old in group A, 69.6 ± 9.2 years old in group B (P > 0.05). There were more female patients in group A (63.5%) than in group B (38.5%). There were more patients who had a history of current cigarette smoking at the time of diagnosis in group A (62.5%) than in B (53.8%). There were 9 patients with combined diseases in group B, but only two in group A. The mean symptomatic period was 18.38 ± 27.12 weeks in group A, but 124.77 ± 142.11 weeks in group B (P < 0.01).

(2) The mean of percentage of ground-glass opacity on HRCT scan was 21.74 ± 11.63 in group A, and 4.67 ± 2.99 in group B (P < 0.001). The mean of percentage of honeycombing on HRCT scan was 3.89 ± 2.98 in group A, and 24.02 ± 11.25 in group B (P < 0.001). The HRCT score was correlated positively with improvement rate in FEV1(r = 0.56, P = 0.008), FVC (r = 0.648, P = 0.001) and TLC (r = 0.492, P = 0.023). The HRCT score correlated negatively with improvement rate in FEV1 (r = -0.585, P = 0.005), FVC (r = -0.85, P = 0.001), DLco (r = -0.476, P = 0.029).

Conclusion Our results showed that the patients with IIP may have good response to corticosteroid treatment if his HRCT score of ground-glass opacity was over 20%, and honeycombing shadow was below 20%. The percentages of ground glass opacity and honeycombing shadow in HRCT before treatment may be an appropriate predictor for the patients with IIP to response to steroid treatment. Conclusion Most IFBs are retrievable by fibreoptic bronchoscopy. Difficulties may occur due to late presentation or to the site/position of IFB. Under GA, bronchoscopy through laryngeal mask airway may make ventilation easier by providing a wider calibre than the endotracheal tube. Backgrounds & Objectives Bronchoscopic intervention in patients with malignant central airway obstruction provides the best ability to maintain airway control as well as initial palliation and stabilization of the airway, allow the possibility of other more effective therapeutic modalities such as surgery, radiation, or chemotherapy. To describe advantage and limitation of emergency rigid bronchoscopic intervention as a bridge to subsequent treatment, we reviewed the medical records of patients who underwent airway intervention because of respiratory failure due to malignant central airway obstruction. Subject & Methods Retrospective review of medical records of 36 patients (26 males, median age 62 years) who underwent emergency airway intervention because of respiratory failure due to malignant central airway obstruction at Samsung Medical Center from January 1999 through December 2003. Results Thirty-four of 36 patients (94.4%) had an improved airway after bronchoscopic intervention. Only one patient required immediate repeat bronchoscopy after the initial successful intervention. After stabilization of airway, twenty-one of 33 patients (58.3%) could have several forms of additional therapy; surgical resection in 9, radiation in 10, combined radiation and chemotherapy in 2 patients. Fifteen patients (41.7%) had no additional therapy. Mean survival time in patients who had additional therapy after emergency rigid bronchoscopy was 38.2 months (range 1.7 to 57.0 months), however, mean survival was 6.2 months (range to 0.1 to 33.7 months) in patients who had no additional therapy. Conclusions Bronchoscopic intervention should be considered even in patients with life threatening respiratory failure due to malignant central airway obstruction for providing opportunity to be performed appropriate therapy for long term survival. Objectives To assess the effects and safety of balloon dilatation using a fiberoptic bronchoscope in the management of benign tracheobronchial stenosis. Methods One hundred and fourteen patients of proximal benign tracheobronchial stenosis were managed by balloon using the flexible fiberoptic bronchoscope. Each patient under-went fiberoptic bronchoscopy, in which a balloon catheter was sent to stenosis segment from the working channel and positioned across the stenosis. Under direct visualization, the balloon was inflated for 1 to 3 min. Repeat inflation-deflation cycles were done if airway narrowing remained after the initial attempt. At preprocedure and last postprocedure days, airway diameters, dyspnea index and complications were evaluated in all of the patients and FEV 1 was tested in 58 cases. Results All of the patients required 1 to 8 sessions, mean 3.22 ± 1.24 sessions. After high pressure balloon dilatations, airway average diameter increased from 2.74 ± 1.32 mm to 6.87 ± 1.76 mm (X ± S) (P < 0.01). Dyspnea index decreased from 2.13 ± 0.81 to 0.71 ± 0.56 (P < 0.01). FEV 1 in the 58 cases's increased from 1.32 ± 0.62 L to 1.81 ± 1.22 L (P < 0.01). No severe complication was found in 114 ptients. Conclusion Airwayplasty of high pressure balloon dilation using flexible fiberoptic bronchoscope is an efficient, safe, simple and rapid method to treat proximal benign tracheobronchial stenosis. Objective To evaluate the efficiency and safety about the bronchial occlusive device of NiTi alloy after being implanted into experimental healthy dogs. Methods The NiTi alloy wire was weaved into varied shapes, such as cast-like, mushroom-like, dumbbell-like and Korean drum-like shapes, and a screw was fixed at the end of it. 24 healthy dogs were divided into four groups and inserted with metal tracheal cannulas. Fibrobronchoscope was inserted through the cannula. After the target location was determined, Guiding wire was sent through the biopsy channel of the bronchoscope, and catheter was then sent along the guiding wire. By the supervision of fluoroscopy and bronchoscopy, implanting device with four kinds of occlusive device at the top was sent respectively into the target location through the catheter of four groups. The changes in the location of occlusive device, bronchial tissue around and in the distal part of it, other key viscera and the occlusive device themselves were determined at the first, third, fifth and eighth week. Results 48 pieces of occlusive device were implanted successfully into dogs at the first time. During eight weeks of observation, 14 pieces of occlusive device (29.16%) displaced or shed. Among them were 2 cast-like, 7 mushroom-like, 5 dumbbell-like device. While all the Korean drum-like occlusive device didn't displaced or shed. Bronchial tissue around the occlusive device was in acute inflammatory reaction at the first week and epithelia began to proliferate at the third week; At the fifth week, epithelia covered the device totally, and distal lung tissue collapsed obviously; at the eighth week, inflammation disappeared and distal lung tissue collapsed totally. There is no obvious obstructive pneumonia in the distal lung tissue during the observation period. After observation, shedding occlusive device was taken out, and no obvious deformation, erosion or rupture was found. Conclusion This bronchial occlusive device of NiTi alloy and its implanting system, which were designed and made in our study, are characterized by accurate localization and easy implantation. In the four different shapes, the Korean drum-like occlusive device has the best effect and satisfactory histocompatibility and we expected it to be used in bronchial closure safely and effectively.

Background To evaluate those factors which might influence diagnostic yield of fiberoptic bronchoscopy (FOB) in evaluating solitary pulmonary nodule (SPN) or lung mass. Method This study was prospectively conducted and performed over a 1-year period in a national medical center. Univariate and multivariate analyses were applied to evaluate those factors. Patients who had pulmonary nodule or mass but no endobronchial lesions underwent FOB with transbonchial biopsy (TBB) and bronchial brushing (BB) under fluoroscopic guidance. Results We studied 69 patients, 52 with malignant tumors and 17 with benign lesions. The diagnostic rate by TBB and BB through FOB for all lesions was 69.6%. There was no significant difference in diagnostic yield between malignant (67.3%) and benign lesions (76.5%). The diagnostic rates of TBB and BB were 50.7% and 52.2%, respectively. TBB and BB were complementary in improving diagnostic rate (P < 0.001). The diagnostic yield of FOB was tightly associated with the size (the diameter > versus £1.8 cm; P = 0.023) and localization (good versus poor; P < 0.001) of the lesion, but not its distance from the tip of FOB. Lesions of the right upper lobe and lesions with no CT bronchus sign were more difficult to be localized and approached by the diagnostic tools. Conclusions We have found that the diagnostic yield of combined TBB and BB through FOB for SPN or lung mass was higher than either one procedure alone. Size of the lesion and good lesion localization during the procedure were important determinants of diagnostic yield in FOB. Good lesion localization was closely related to the location and the presence of CT bronchus sign of the lesion. Alternative diagnostic approaches should be considered for lesions under 1.8 cm in diameter that are located in the right upper lobe with no CT bronchus sign. Background Obstruction of the airways due to malignant disease is a frightening condition that portends poor prognosis. Emergency airway stenting can quickly palliate and relieve the obstruction. We retrospectively analysed our data on the management of these patients. Methods From 1999-2004, records of 11 patients with urgent tracheobronchial stenting using rigid bronchoscopy for palliative relief of airway obstruction.

The median age of these 11 patients (6 males and 5 females) was 43 yrs (range 18-69 yrs). The diagnoses were as follows: NSCLC (7), adenoid cystic carcinoma (1), small cell lung cancer (1), Hodgkin's lymphoma (1) and malignant thymoma (1). A total of 15 airway stents were inserted consisting of 2 tracheal stents, 7 left bronchial stents and 6 right bronchial stents (6 covered/5 uncovered ultraflex stents and 4 polyflex stents). All patients had immediate symptomatic relief. Follow up is as follows: 6 patients died within a year of stent insertion from their underlying malignancy, 2 were transferred back to their respective countries and were lost to follow up, the 3 remaining patients are still being follow up by the oncologist. The patient with Hodgkin's lymphoma had regression of her disease with chemotherapy and is well 2.5 years after stent placement. Conclusion Malignant airway obstruction from extrinsic or intrinsic causes can be managed by urgent palliative airway stenting, which provide instant relief in an otherwise fatal condition. Their outcome however is still poor and is determined by their underlying malignant disease. Background Hypoxia is an acknowledged feature of most solid tumor. The ability of tumors to adapt to a hypoxic microenvironment is increasingly recognized as an important mechanism that promotes tumor growth. Hypoxia inducible factor-1(HIF-1) is a transcriptional factor that is important in regulating the expression of various genes that are overexpressed in the presence of cellular hypoxia. These genes include the hematopoietic growth factor erythropoietin, angiogenic growth factors, and genes involved in glucose transport and metabolism. HIF-1 is a heterodimer composed of HIF-1a and HIF-1b subunits. HIF-1b (also known as the aryl hydrocarbon receptor nuclear translocator) is a common subunit of multiple BHLH-PAS proteins, whereas HIF-1a is unique, O 2 -regulated subunit that determines HIF-1 activity. Despite intense investigation of HIF-1 in human common tumors, little has been learned about the effects of HIF-1 on lung cancer. Methods 1. Immunohistochemistry (IHC) technique was employed to detect the protein expressions of HIF-1a, P53 and vascular endothelial growth factor (VEGF) in specimens from 57 patients with lung cancer. 2. To sift the A549 cells which can express HIF-1a stablely, HIF-1a mRNA was amplified by RT-PCR and inserted into plasmid pcDNA3. A549 cells were sifted with G418, after the expression plasmid pcDNA3/HIF-1a transfected into A549 through Lipofect2000. 3. The effect of HIF-1a on tumor growth in vitro was evaluated by the growth curve and subcutaneous implanted tumors of nude mice in vivo. A549 cells (1 ¥ 10 6 /mouse) were inoculated subcutaneously into 20 nude mice, which were randomly divided into two groups: the control group (group A, n = 10), the HIF-1a transfected group (group B, n = 10). The weights of subcutaneous tumor were detected. The resected specimens were made into paraffinembedded sections. The Proliferating Cell Nuclear Antigen (PCNA) was identified by IHC. The expressions of HIF-1a, apoptosis-related protein survivin and bcl-2 were analyzed by Western-blot. 4. To evaluate HIF-1a on tumor chemotherapy resistance, 5-Fu was added into A549 after HIF-1a transfected, the growth activity was measured by growth curve, apoptosis was detected by flow cytometry, at 48 h, the levels of caspase3 and mdr1 were found by Western-blot. 5. To study the role and mechanisms of HIF-1a silence on the growth and tumorigenicity of lung cancer cells A549, the antisense oligonucleotide of HIF-1a was transfected to A549 cells. The effect of the antisense oligonucleotide on tumor growth in vitro and in vivo was evaluated by the growth rate suppression of A549 cells and subcutaneous implanted tumor in nude mice, and the effect on tumorigenicity was evaluated by the expression inhibition of angiogenic factors, the microvessel density (MVD) and VEGF protein expression which were detected by IHC and Western blot respectively. Results 1. The results indicated that the total positive proportion of HIF-1a expression was 63% and the HIF-1a expression was more frequent in bronchiole-alveolar carcinoma (86%) than in other lung cancer.

The was a strong association of HIF-1a with VEGF and P53 protein expressions. 2. The constructed expression plasmid was analyzed with restriction enzymes and gel electrophoresis. Two DNA lanes at 5.3 kb and 2.55 kb, respectively was found, which is consistent with what expected. The expression plasmid was transfected into cultured A549 cells through Lipofect2000, the expression of HIF-1a protein detected by Western-blot was significant increased. 3. The growth rates of the HIF-1a transfected lung cancer cells A549 were significantly increased, and more importantly, the HIF-1a transfected lung cancer cells A549 were able to enhance lung cancer growth in nude mice (P < 0.05). The PCNA were increased significantly in group B when compared with group A. The expressions of HIF-1a, survivin and bcl-2 in group B were increased significantly than that of group A. 4. The growth rate of 5-Fu group was inhibited and the apoptosis index and caspase3 activity were found increased significantly when compared with control group. After HIF-1a transfected into A549, the apoptosis index and the activity of mdr1 was decreased, the effect of 5-Fu was weakened, which indicated HIF-1a enhance the chemotherapy resistance. 5. This study revealed that in vitro the growth rate of antisense oligonucleotide group was significantly decreased as compared with that of control group, sense oligonucleotide group and falsesense oligonucleotide group, in vivo the weight of implanted tumors on nude mice of antisense oligonucleotide group was 1.51 ± 0.40 g, which was significantly lower than that of control group (2.79 ± 0.33 g), sense oligonucleotide group (2.81 ± 0.45 g) and false-sense oligonucleotide group (2.89 ± 0.39 g) and the inhibitory rate was 47%. Both MVD and VEGF protein expression were significantly inhibited in antisense oligonucleotide group compared with those in other groups.

Conclusions HIF-1a overexpression is a common event in lung cancer, which is related to the upregulated of the angiogenic factor VEGF and oncogene mutant P53 protein. HIF-1a could increase lung cancer cells A549 growth and promote chemoresistance, its mechanism may be due to the fact that it can enhance proliferation and inhibit apoptosis. The antisense oligonucleotide of HIF-1a could inhibit lung cancer cells tumorigenesis by inhibiting vasculars growth in vitro and in vivo. There was 1 case out of 9 cases of cavities belonged to multiple cavities; 1 case belonged to clean cavities and all the other 7 cases were tuberculofibrosis cavities. The diameter of calcification was 0.5 cm-2.5 cm, single or multi-located in different foci from most tumour. The size of yellow tubercle was about 0.5 cm-2.0 cm. They were single, multipul and numerous tiny nodules in whole lung parenchyma, considerably varying in size. There 46 cases telling that yellow tubercle and cancer could coexist in the same zone or stayed around the cancer.

Conclusion From the analysis result of the study, it could be concluded that pulmonary tuberculosis is a dangerous factor of lung cancer, especially for the patients who have more than 20 years disease history. The chronic injure of pulmonary tuberculosis to the lung affects the function of bronchial epithelium and the immune function of economy aganist pulmonary tumors. It even indirectly accelerates the formation of tumour. It is the calfication of tuberculosis, the formation of scar tissue, the hyperplasia of fibrous connective tissue, the hyperplasia of bronchial epithelium, the hyperplasia of bronchial and alveolus epithelium and the dysplasia that has the certain cause and effect relation with the formation and development of lung cancer.

In the future, the study on exploring the pathogenesis of the co-existence of pulmonary tuberculosis and lung cancer will be continued by the valuable data and using molecular pathologic technic. Background Nuclear factor-kappa B (NF-kB) was first identified as a B-cell nuclear factor and given its name on the basis of its ability to bind to an intronic enhancer of immunoglobulin k-light chain gene. Since then, NF-kB has been identified in numerous cell types and is found to be activated by a wide range of inducers, including ultraviolet irradiation, cytokines, inhaled occupational particles, and bacterial or viral products. In resting cells, NF-kB resides in the cytoplasm in an inactive from bound to an inhibitory protein known as IkB. Upon cellular activation by extracellular stimuli. IkB is phosphorylated and proteolytically degraded or processed by proteasomes and other proteases. This proteolytic process activates NF-kB, which then translocates into the nucleus. In nuclei, NF-kB can initiate or regulate early-response gene transcription by binding to decameric motifs, 'GGGRNNYYCC (kB motif)', found in the promoter or enhancer regions of specific genes. NF-kB binding sites have been identified in the promoter regions of genes whose products are intimately involved in tumor cells apoptosis and proliferation. We hypothesized that the transfection of a sufficient quantity of decoy oligodeoxynucleotides (ODN) containing the NF-kB cis element into lung cancer cells A549 would effectively bind NF-kB, thus influence A549 cells growth and apoptosis. Methods First, the lung cancer cells (A549) were divided into three groups: group A (control group); group B (decoy ODN group) and group C (scramble decoy ODN group). FITC-labeled NF-kB decoy ODN was transfected into A549 with LipofectAMINE TM 2000, the activation was observed by electrophoretic mobility shift assays (EMSA), the proliferation was observed by growth curve, the apoptosis of cells were observed by flow cytometry and TdT mediated dUTP-biotin Nick End Labeling (TUNEL), the expression of bcl-2 and fas were observed by Western blot. Second, When ciglitazone was added, the proliferation was observed by growth curve and the differentiation of cells was observed by flow cytometry, the expressions of cyclinD1 and MDR1 were observed by Western blot.

Results After FITC-labeled decoy ODN was transfected for 1 hour, the decoy ODN was detected in the nuclei of A549 cells, EMSA performed the depression of the NF-kB binding to the nucleus. The growth curve showed the inhibition of the growth of A549, the percentage of apoptosis was increased compare with control group by flow cytometry and TUNEL. The amount of apoptosis inhibitor (bcl-2) in group A and group C were 2.0 times and 2.1 times more than that of group B respectively. The lever of apoptosis accelerator (fas) in group B were 2.6 times and 3.3 times more than that of group A and group C espectively via Western blot. At the same time, western blot showed the decoy ONDS could decrease the lever of mdr1. Ciglitazone could obviously inhibit the growth and induce the differentiation of A549 cells that were transfected with NF-kB decoy ODNs. There were more cells arrested in G 1 /G 0 phase and the expression of cyclinD1 was markedly down-regulated than in control cells.

Conclusions The NF-kB decoy ODN can accelerate the apoptosis of lung cancer cells A549 and its mechanism may be due to the fact that it can inhibit the expression of bcl-2 and increase the lever of fas. At the same time, NF-kB decoy ODNs could enhance the effects of ciglitazone on proliferative suppression and differential induction of A549 cells, which suggested NF-kB decoy ODNs could have synergistic action with chemotherapy drug ciglitazone.

Background and Aim Tenascin (Tn)-C is an extracellular matrix protein that is involved in tissue interactions during fetal development and oncogenesis. To determine the functional role of serum Tn-C in non-small cell lung cancer (NSCLC), we measured serum levels of Tn-C as well as other factors implicated in angiogenesis, and sought these relationships. Material and Methods A total of 68 patients with a primary NSCLC who underwent surgical operation were analyzed in this study whose mean age was 65 ± 10 years. Adenocarcinoma was diagnosed in 43 patients, squamous cell carcinoma in 15 patients, and other histology in 5 patients. As control, 47 aged-matched healthy volunteers were employed. Serum Tn-C, Vascular endothelial growth factor (VEGF), and osteopontin (OPN) levels were measured with commercial sandwich ELISA kits. Intratumoral Tn-C expression and microvessel density (MVD) were evaluated with immunohistochemical analysis.

Results A mean concentrations of serum Tn-C in all NSCLC patients was slightly higher than that of normal controls, but this difference was not statistically significant (91 ± 6.7 ng/ml vs. 75 ± 4.7, P = 0.079). A positive correlation between serum Tn-C levels and MVD (r = 0.325, P = 0.009), serum OPN (r = 0.416, P = 0.0005), VEGF (r = 0.48, P < 0.0001), and SCC levels (r = 0.356, P = 0.0029) was observed. In contrast, there was no correlation between serum Tn-C levels and serum MMP-9 levels (r = 0.097, P = 0.44), serum CEA (r = 0.099, P = 0.422) and SLX levels (r = 0.007, P = 0.957).

Intratumoral Tn-C expression was co-localized with microvessels in the stroma but not in cancer cells by immunohistochemical analysis. Interestingly, human umbilical vascular endothelial cell (HUVEC) migrated toward recombinant Tn-C by Boyden chamber's modified method.

Conclusion These results suggest that Tn-C may play an important role in angiogenesis of patients with NSCLC, and measurement of serum Tn-C may be helpful in predicting intratumoral vasculature.

Background To evaluate the prognostic value of vascular endothelial growth factor (VEGF) in patients with lung cancer.

Methods VEGF level was measured by sandwich ELISA in 112 cases with lung cancer. Those were also measured respectively that was 48 cases diseased control group and 31 cases healthy individuals. That else included 46 cases of malignant effusion and 31 cases of tuberculosis pleural effusion. Results The serum, urine and pleural effusion VEGF levels in patients with lung cancer were 506.22 ± 365.06 mg/ml, 167.40 ± 186.89 mg/ml and 937.23 ± 400.92 mg/ml respectively, which were significantly higher than those of diseased control groups and healthy groups. The serum and urine VEGF level in patients with lung cancer were significantly higher in advanced stage of the disease (P < 0.001) than those in early stage. As to the degree of invasion (T factor), the serum and urine VEGF levels of T4 invasive lesions were significantly higher than those with T1-T3 invasion (P < 0.001). The same was also true in patients with lung cancer having lymph node metastasis N2-N3 than that without lymph node metastasis N0 (P < 0.001). The pleural effusion VEGF level in lung cancer patients was significantly higher than those with tuberculosis effusion, and also higher than those in serum (P < 0.001).

Conclusion VEGF plays an important role in the growth and metastasis of lung cancer. VEGF in patients with lung cancer is an independent prognostic factor and may be proposed for use in the clinic and research. Au: Please provide all authors' names

Background Drug resistance remains the thorniest obstacle in lung cancer chemotherapy. Especially the acquired drug resistance induced by commonly used antitumor drugs influences the prognosis severely. It is commonly considered that p53 is the highest related gene to inherent drug resistance of NSCLC. But the relationship between p53 and the acquired drug resistance induced by anti-tumor drugs is not clear.

The model of a multi-drug resistance human large cell lung cancer cell line (H460/cDDP) was established by high dose cisplatin intermittent selection from the parental cell line H460. Drug sensitivity was determined by MTT assay; Several multidrug resistant related proteins including P-gp, MRP-1, LRP, GST-p, TOPOII and P53 were checked by immunocytotochemistry. P53 protein was detected by Western-blot. Immunofluorescence was used to assess the extent of P53 phosphoration. p53cDNA from H460/cDDP was amplified and sequenced, then compared with the wild type p53 from H460 cells. Amino acid sequence and the structure of the mutated P53 protein were predicted by bioinformation methods (Gene Tool and Swiss-PDB viewer softwares). 96 genes at p53 signaling pathway were detected by gene chip array. H460/cDDP was transfected by the Pshuttle-CMV-wtP3 plasmide. MTT assay was used for evaluating the efficacy of the transfected wtp53 on the drug sensitivity. H460/cDDP and the sensitive parental cells H460 were transplanted subcutaneously to nude mice. A recombination of adenovirus and wild-type p53 (Ad-p53) alone or combining with cisplatin was injected to nude mice to evaluate the anti-tumor and their reversing effect of drug resistance. Results Drug-resistant H460/cDDP cell line developed after 6 months of cisplatin selection. Resistance indices to cisplatin and carboplatin in H460/cDDP cell line were 10.21 and 9.98 respectively. At the same time, the cell line also exhibited moderate resistance to 5-Fuorouracil, etoposide, methotrexate, adriamycin, epirubicin, bleomycin and navelbine, except for taxol. P53 protein was found positive in the H460/cDDP cell but translocated from nuclear to cytoplasm. The velocity of P53 protein electrophoresis varied from that of H460 and showed no phosphoration by cisplatin stimulation. In addition, there was stronger expression of LRP but not other protein in H460/cDDP cells. Sequencing of p53cDNA of H460/cDDP showed an insertion 't' after site 277. The mutated P53 protein, with 39 amino acids lost in the N-terminal, was coded from site 370 for the insert 't' after site 277, resulting to the loss of the MDM2 binding site and Serine15 phosphorylation site. Gene chip array for 96 related genes at the p53 signaling pathway showed that P53AIP1, c-myc, PKCa, PKCb, and 6 other genes were expressed in H460/cDDP, but not in its parental cells. MDM2, NUMB and EP300 that regulating p53 expression increased by 4-8 folds in the multi-drug resistant cells. p53 modification genes KIP2 and MAP2K4 were also 10 folds up-regulated also. As an apoptosis regulator and cell-cycle controller at the downstream of p53, LRDD expressed 10.89 folds higher in H460/cDDP. DNA repair gene GADD45A was 4.68-fold higher. There were few genes such as ABCB1, CASP9 and TNFALP1 appeared to be down-regulated in the drug resistant cells, but showed no significant difference as compared with the parental cells. The resistance to cisplatin was partly reversed (53.2%) when H460/cDDP cells were transfected with the Pshuttle-CMV-wtP3 plasmide. With regards to the effect on the tumor grafts, Ad-p53 in combining with the cisplatin demonstrated better inhibition effect than either one single administration. The inhibition effect on the multi-drug resistant cells was about the same as that on the sensitive parental cells. 

T KASAI 1 We observed 4 partial response (15.4%) and 9 disease stabilizations (NC 34.6%) lasting at least 2 months, for an overall disease control rate of 50%. The mean time of follow-up was 6.9 months (range: 0.9-15.9 months), The median time to progression (TTP) was 6.7 months ± 1.5 months (95%CI: 3.8-9.5), including 18 patients (69.2%) who had TTP of 6 months or longer. By the date of cut-off, 70.4% (19/27) patients still alive with median overall survival time (ST) 7.5 months ± 1.2 months, (95%CI: 5.1-9.9). The side effects were generally mild and consisted mainly of diarrhea and skin toxicity. Grade 1-2 diarrhea occurred in 4 patients (15.4%) and 2 patients (7.6%) experienced grade 3/4 diarrhea requiring hospitalization with 1 patient (3.8%) withdrew from the study. Grade 1-2 skin toxicity including rash, pruritus, dry skin, acne, occurred in 11 patients (42.3%). Conclusion Gefitinib is safe and well tolerated in elderly NSCLC patients. The disease-control rate achieved suggests that this drug could represent a valid option in the management of this unfavorable subgroup of patients.

Background 2-(1-hydroxyethyl)-5(or 8) hydroxynaphtho[2,3-b]furan-4,9dione (FNQ 3 ), a phytochemical analog of furanonaphthoquinone compounds, is suggested to be cytotoxic to cancer cells. The objective is to study the relationship between the antitumor activities of FNQ 3 and Fas/FasL systems. Methods To study the dose-dependent effect, the different dose of FNQ3 was added to A549 cells for 24 hs, the Fas mRNA was detected by QRT-PCR and the cell surface Fas protein expression and apoptosis were analyzed using flow cytometry. To study the time-dependent effect, 0.5 ug/ml FNQ 3 was added to A549 cells for 1 day, 2 days, 3 days, 7 days, the Fas mRNA was detected by QRT-PCR and the cell surface Fas protein expression and apoptosis were analyzed using flow cytometry. Results Along with the increased dose of FNQ 3 , the Fas mRNA expression, cell surface Fas protein expression, the sensitivity of apoptosis were increased in A549 cells. As the concentrate of FNQ 3 was 2.5 ug/ml, the Fas mRNA PCR products was 75000 molecules/ul and the apoptotic cells (%) was 37% (P < 0.001). As well as the time-dependent effect, the Fas mRNA expression, cell surface Fas protein expression, the sensitivity of apoptosis were increased along with the prolonged time. When FNQ 3 (0.5 ug/ml) was added to A549 cells for 7 days, the Fas mRNA PCR products was 150 000 molecules/ul and the apoptotic cells (%) was more than 30% (P < 0.02).

Conclusion FNQ 3 as a kind of anti-tumor medicine can increase the cell surface Fas protein expression and up-regulate the sensitivity of Fasmediated apoptosis. The up-regulation was both time-and dose-dependent. Backgorund and Objectives Cyclooxygenase-2 (COX-2) is found to express in many solid tumors and may play an important role in pathogenesis of cancer. To investigate relationships between cyclooxygenase-2 (COX-2) expression and angiogenesis in non-small cell lung cancer (NSCLC). Methods Special antisense for COX-2 was constructed into p31 plasmid and transfected stably H1299 cells (H1299-COX-2). mRNA and protein of COX-2 and vascular endothelial growth factor were determined with the method of RT-PCR in H1299 cell lines and H1299-COX-2 cell lines and Western bloting. The expressions of Vascular endothelial growth factor, COX-2 and CD31 were detected with immunohisytochemical staining. The relationships between the parameters and between both parameters and prognosis of the patients were determined.

Results COX-2 mRNA and protein in H1299-COX-2 cell lines were significantly lower than in the H1299. In NSCLC, expression of COX-2 was related to expressions of Vascular endothelial growth factor and CD31 (P < 0.05 Transforming growth factor (TGF)-b 1 is known to regulate many cellular processes, including cell proliferation, differentiation, and apoptosis. Plasma TGF-b 1 levels has been shown to be elevated in patients with lung cancer.

In this study, we test the association of the polymorphisms at 2 loci, i.e. 

Objective To study the expression of survivin gene and its relationship with prognosis in non-small-cell lung cancer (NSCLC). Methods Envision immunohistochemisty was used to detect the expression of survivin in 60 cases of NSCLC tumor samples and 8 cases normal tissue.

Results There was no expression in normal tissue, and expression of the survivin gene in 61.6% (37/60) NSCLC. There was significant relationship between survivin gene expression and prognosis of NSCLC (P < 0.05).

The result indicate that, in NSCLC, survivin gene expression may be a useful prognostic factor.

Background Lung cancer remains the world-wide leading cause of cancer morbidity and mortality as a result of late presentation or diagnosis. Surgical resection could be curative for early stage lung cancer while prognosis for advanced stage lung cancer is still poor despite continuous development and trial of new therapeutic strategies and chemotherapeutic agents. Study in lung carcinogenesis is essential for better understanding of lung cancer biology that may open up new diagnostic and therapeutic targets for intervention. Gene expression represents the underlying genetic changes and reflects the activities of cancer cells. Microarray is a high throughput method that allows for parallel study of the expression of thousands of genes, which will allow for gene expression profiling and class discovery. Our hypothesis is that differential gene expression in lung adenocarcinomas could be correlated with clinical characteristics and molecular classification of tumour subtypes.

Methods We have collected resected lung adenocarcinomas (n = 49) and normal lung tissue (n = 9). Total RNA from these specimens was processed according to standard procotol from Affymetrix and then hybridized onto Affymetrix GeneChip HG-U133A with one sample per GeneChip. Data was analysed with dChip 1.3 and correlated with clinical characteristics. Results Differential gene expression was obtained with two sample t-tests of log transformed signal intensities, yielding 318 genes having up-or downregulation by four fold difference, and some of these can be classified according to their known gene functions such as involvement in cell cycle or signal transduction. Unsupervised data analysis with hierarchical clustering aiming at detection of co-regulated or co-expressed genes showed significant gene clustering of molecules involved in cell cycle and proliferation, namely TOP2A, CENPF and MCM2 (P = 0.008); as well as consistent sample clustering of normal lung tissues and groups of adenocarcinomas. Supervised data analysis with principal component analysis were applied for class discovery, with a group of 35 filtered genes being used as potential molecular classifier for subtypes of adenocarcinomas. Further correlation with clinical characteristics and prognosis will be analysed.

We have identified differentially-expressed genes in lung adenocarcinomas of potential biological significance that deserve further investigation as potential diagnostic or therapeutic targets in the management of lung cancer. Background Lung cancer is the leading cause of deaths from cancer in Japan and world wide. In the recent analysis of the response to the EGFR tyrosin kinase inhibitor gefitinib (Iressa TM ), somatic mutations in the tyrosine kinase domain of the EGFR gene were found in patients with gefitinib-responsive lung cancer. However, no prospective study to search for the correlation between the mutations and the gefitinib-responses has been performed. Methods Tumor specimens were obtained during diagnostic or surgical procedures, including trans-bronchoscopy lung biopsy, metastatic bone tumor biopsy, thoracentesis cells of malignant pleural effusion, and surgical lung tumor samples. Mutations in the EGFR gene (exons 17 to 24) in tumors from patients with NSCLC were searched. Results Mutations were identified in 2 of 11 patients (2 squamous cell carcinoma and 9 adenocarcinoma), one of which was 15 bp deletion in exon 19 and the other was L858R in exon 21. Identical mutations from both primary and metastatic sites (one from pleural effusion cells and the other from bone metastatic tumor) were detected in the same patients. Both cases harboring mutations were female adenocaricinoma, which was consistent with the characteristics of reported gefitinibsensitive patients. The oral gefitinib administration revealed dramatic responses to these mutation-positive patients.

Conclusion EGFR mutation analysis was successfully performed from the tumor samples not only by surgery but also by daily trans-bronchoscopic tumor biopsy and by thoracentesis of malignant pleural effusion. EGFR mutations in the catalytic domain can be a reliable predictor of the responsiveness to gefitinib and will lead to the first personalized medicine in solid tumors.

MINGWEI CHEN, YU YAO, ZHIHONG SHI

Objective As a tetra-Anulus triterpene Saponin extracted from ginseng, ginsenoside Rg3, was first found by a Japanese scholar [1] to have a selective function of inhibiting the infiltration and metastasis of tumor cells. The aim of the study was to make a further evaluation of Ginsenoside Rg3 in clinical. Purpose Tumor cell expression of COX-2 has been implicated in the progression of murine and human lung cancer. Inhibition of COX-2 by nonsteroidal anti-inflammatory drugs reduces the risk of cancer development in humans and suppresses tumor growth in animal models. However, the underlying mechanisms for this beneficial effect are not fully understood. In this study we explore the potential link between the anti-cancer effects of COX-2 inhibitors and the expression of the integrin a5b1. Experimental design Using human non-small cell lung carcinoma cell line, we assayed for integrin a5 mRNA and protein expression by RT-PCR, real-time RT-PCR and Western blot analysis. phosphorylation of extracellular signal-regulated kinase (Erk) measured by Western Blot analyis. Nuclear protein binding activities to three response elements located in the a5 promoter [CCAAT/enhancer-binding protein (C/EBP), Specificity protein 1(Sp1) and Activator protein-1(AP-1)] were measured by gel shift mobility assay. We used transient transfection assays with a5 promoter reporter gene constructs to determine the regulatory sites in this promoter, which mediates COX-2 inhibitors regulation of a5.

We found that the selective COX-2 inhibitors NS398 and Nimesulide decreased mRNA expression and protein production of the a5 subunit of the integrin. This effect was associated with inhibition of NSCLC cell adhesion to fibronectin. The COX-2 inhibitors triggered the phosphorylation of extracellular signal-regulated kinase (Erk) in a time-dependent manner, and the inhibitor of Erk PD98095 prevented their inhibitory effects on a5 expression. Transient transfection assays showed that the COX-2 inhibitors affected a5 gene transcription by acting between -92 to-41 bp of the human a5 gene promoter. Gel mobility shift assays showed that the COX-2 inhibitors increased Sp1 DNA binding, but decreased that of AP-1. These effects were accompanied by an increase in Sp1 protein and a decrease in c-Jun protein expression, as well as inhibition of SAPK/JNK phosphorylation. The Sp1 inhibitor, Mithramycin A, also blocked the inhibitory effect of the COX-2 inhibitors on a5 expression and promoter activity. Conclusion These findings suggest that COX-2 inhibitors suppress a5b1 integrin expression in NSCLC through effects on a5 gene transcription mediated by Erk activation, increased Sp1, decreased AP-1 DNA binding, and inactivation of SAPK/JNK signals. Together, our observations unveil a new mechanism of action against NSCLC for COX-2 inhibitors that relates to regulation of a5 gene expression and, consequently, recognition of extracellular matrices (i.e., fibronectin) by tumor cells. Background Tumor stage and its histological subtype remain the most important predictors of clinical behavior in current pulmonary practice of lung cancer. However, many investigators agree that these parameters are not sufficient to predict which tumor will recur, even after radical curative surgery. Therefore, it is necessary to evaluate the significance of other morphological, biological and molecular parameters beyond TNM classification.

Objective Occult micrometastasis to mediastinal lymph node, which could not be detected by routinely histopathologic examination, might be correlated with the prognosis of patients with non-small cell lung carcinoma (NSCLC). Molecular biologic staging is the assessment of tumor markers that are assessed to detect the presence of occult metastases in lymph nodes to improve the risk stratification provided by comventional TNM stageing. The aim of this study was to diagnose occult micrometastasis of mediastinal lymph node in NSCLC patients and assess molecular biologic staging and evaluate its prognostic significance. Conclusion CEAmRNA, P53 and AE1/AE3 expression provide prognostic information, allowing a more accurate outcome prediction for the individual patient, but were not independent prognostic factors. The value of lymph node micrometastasis and molecular biologic substaging for prognosis of non-small cell lung cancer is undefined. The best prognostic index for operable non-small cell lung cancer (NSCLC) is the TNM staging system. Molecular biology holds the promise of predicting outcome for the individual patient. These data suggest that molecular analysis allows a more accurate assessment of staging. However, larger studies are needed to determine the clinical role of molecular staging.

Background The multi-drug resistance (MDR) is one of the major causes of failure of chemotherapy. It's necessary to explore the efficacy of dihydroartemisinin on MDR reversal and its mechanism.

Methods Human lung adenocarcinoma cell line A549 and its cisplatin-resistant countepart A549/CDDP were observed in this study. The inhibitory effects on proliferation were measured by MTT assay. The effects of dihydroartemisinin on expression of Bcl-2 and Bax were detected by histochemstry. Cell apoptosis was detected by flow cytometry after annexin-V and PI dual labeling.

The IC 50 of cisplatin in A549 and A549/CDDP was 0.351 mg/ml and 5.703 mg/ml, The drug resistance multiple was 16.24. In the dihydroartemisinin plus cisplatin group, the IC 50 of cisplatin was 2.225 mg/ml in the low dose group and 0.464 mg/ml in the high dose dihydroartemisinin group. The reversion multiple was 2.53 and 12.29. Detected by histochemical method, the expression of Bcl-2 decreased and that of Bax increased after the dihydroartemisinin treatment on A549/CDDP cells. The apoptosis rates were 11.5% in the dihydroartemisinin group and 0.4% in the control group.

Conclusion The resistance of cisplatin by A549/CDDP was related with apoptosis resistance. Dihydroartemisinin could reverse cisplatin resistance by inducing cell apoptosis.

Background There is many measures which doctor can utilize in the diagnosis of Lung cancer. But sometimes, when such measures as radiography, brochoscopy, and biopsy are carried out, we still can not confirm a diagnose. P53 and K-ras are considered as the molecular marker of lung cancer. The study try find another method on diagnosis of lung cancer.

Objectives To investigate the rate of p53 and K-ras gene mutations in sputum of non-small-cell lung cancer (NSCLC) and estimate its value. Subject and Methods 42 cases of diffenent stage NSCLC patients who were proved by pathology were involved in study group. These patients include: 20 cases squamous cell carcinomous and 22 cases adenocarcinomous. 13 cases of patients with COPD or other benign disease were involved in control group. Sputum from these patients was detected with PCR-SSCP technique for p53 and K-ras gene mutation.

The mutation rate of p53 in NSCLC and control group is 40.4% and 15.4% respectively (P < 0.05), K-ras is 26.1% and 7.69% respectively (P < 0.05). The stimutaneous mutations is 11.9% and 0 (P < 0.05). There is no significant difference between smoking patients and no-smoking patients. Chemotherapy have no influence to p53 and K-ras mutations.

Conclusions The NSCLC patients have the higher mutations rate than control group. The combined detection for p53 and K-ras mutation detection in sputum have high specificity to NSCLC. It is an nonvansive method to detect them in sputum and is tolerable to patients.

KUN LI, XIAO MEI QIN 1 , SHU HONG LIU 1 ICU, 1 

Background A lot of laboratory experiments and epidemiologic studies have suggested that the consumption of green tea may reduce the risk of a variety of human cancers. The main chemical components of green tea are green tea polyphenols (GTP), which are responsible for the anticancer activity of green tea. There are many mechanisms concerning GTP's effects, but none of them could completely explain GTP's wide anticancer activities. It is well known that the growths of all solid tumors depend on angiogenesis, without neovascularization, the primary tumor would not reach the size of 1-2 mm 3 . GTP's antiangiogenesis activity has been demonstrated in vitro. So it is reasonable to surmise that GTP may inhibit tumor angiogenesis in vivo. Methods Lewis lung carcinoma was employed as the model to study the antiangiogenesis activity of GTP by using the methods of immunohistochemstry. C57BL/6J mouse were divided into six groups randomly, two control groups and four treatment groups. The latter were orally administered GTP 62.5, 125, 250, 125 mg/kg d -1 respectively 24 hours after implantation. At day 13 and 23, mouse were killed seperately to study tumors' angiogenesis and metastasis. VEGF, C-Jun, PCNA and MVD were measured immunohistochemically. The metastatic foci on the lung surface were counted under a magnifying glass.

Results Lewis lung carcinomas were greatly inhibited, with an inhibitory rate of 27.2% by GTP (125 mg/kg d -) and MVDS were decreased by 29.5% correspondingly. GTP also lowered the expression of VEGF, PCNA and C-Jun, the inhibitory rates were 53.8%, 15.6%, and 12% respectively. The average number of metastatic foci on lung surface was decreased from 13.17 to 5.17 after three-week treatment of GTP (125 mg/kg d -).

Conclusion GTP may exert at least part of its anticancer effect by inhibiting angiogenesis through blocking the expression of C-Jun and VEGF. 7%) . In all cases, the lesion presented as an intra-pulmonary mass or nodule. The mass density was almost inhomogeneous. Calcification was seen in one case (4.5%) and cavity was seen in 5 cases (22.7%). A triangular shape of lesions was demonstrated in 3 cases (13.7%), round-like shape in 19 (86.3%), calabash-like shape and strip-like shape in one case each. The mass showed a lobulated border in 19 cases (86.3%), coarse spicules in 7 patients (31.8%), and pleural indentation sign in 3 cases (13.7%). Bronchial stenosis was found in 3 cases (13.7%), of which obstructive atelectasis was accompanied. Encasement of great vessels in mediastinum and concomitant pleura or thoracic wall occurred in one case and in 12 cases (54.5%). Hilar and mediastinal lymphadenopathy was seen in 15 (68.1%) and in 14 (63.6%). Metastases were found in the same lobe of lung (n = 2) and bone (n = 1). Conclusion Pulmonary carcinosarcoma is mainly of peripheral type. On CT scan, the tumor has inhomogeneous density. Pulmonary carcinosarcoma is apt to invade concomitant pleura or thoracic wall, and both hilar and mediastinal lymph nodes. Background Cheyne-Stokes respiration with central sleep apnea (CSR-CSA) has been detected in as many as 50% of patients with chronic congestive heart failure (CHF). The presence of CSR-CSA in a patient with CHF confers a worse prognosis. Unfortunately, there are few data on treating CSR-CSA in patients with CHF. The plasma concentration of several neurohormones has been shown to be independent markers of left ventricular ejection fraction (LVEF) and mortality in patients with heart failure. Our study was to compare the effects of assisted ventilation (Autoset CS, ResMed, Australia) on clinical outcomes and plasma BNP concentrations in elderly patients with heart failure and CSR-CSA.

Methods 25 consecutive patients with stable heart failure were underwent overnight polysomnographic study, echocardiographic evaluation and 6 minute walk test before and after treatment. In addition to standard therapy, patients with CSR-CSA (n = 16) were treated by Autoset CS for 4 weeks. Plasma BNP concentrations were measured by radioimmunoassay.

Results Plasma BNP levels were significantly greater in patients with CSR-CSA than in patients without CSR-CSA [458.6 ± 228.6 ng/L (n = 16) vs 245.4 ± 212.4 ng/L (n = 9), P < 0.01]. Plasma BNP levels were negatively correlated with LVEF (r = -0.78, P < 0.01) and 6 minute walk distance (r = -0.58, P < 0.05). There was positive correlation between BNP and central apnea-hypopnea index (AHI, r = 0.64, P < 0.05) in patients with CSR-CSA. Autoset CS treatment was associated with decrease of plasma BNP levels (458.6 ± 228.6 ng/L to 245.4 ± 181.4 ng/L, P < 0.05) and improvement of central AHI (31.5 ± 9.6 per hour to 10.1 ± 7.6 per hour, P < 0.001), LVEF (0.29 ± 0.18 to 0.38 ± 0.22, P < 0.05) and exercise capacity (220 ± 114 meters to 332 ± 121 meters, P < 0.01).

Conclusion CSR-CSA is also common in elderly heart failure patients. Autoset CS is effective in treating CSR-CSA in heart failure patients according to clinical and neurohormonal changes.

Clinical implications CSR-CSA should be a target for current heart failure treatment strategies. BNP plasma levels could reflect the level of therapeutic success in patients with heart failure.

Background During noninvasive ventilation in COPD patient, the difference in patient's response to pressure support ventilation (PSV) and proportional assist ventilation (PAV) was uncertain. A self-control cross-over study was conducted to compare the clinical and physiological response to PSV and PAV in COPD.

Methods 9 severe COPD patients recovering from acute exacerbation were recruited for the study. The experiment was consisted of spontaneous breathing and 6 ventilatory support conditions with two modes and three levels of ventilatory support. Esophageal, gastric, airway opening pressures and flow were measured simultaneously. Transdiaphragmatic pressure (pdi), pressure-time product (PTP) of the inspiratory muscle and work of breathing (WOB) were calculated. Results There were significant reduction in average Pdi, PTP and WOB performed by the patient (Wi, p) during PSV and PAV compared with SB, the reduction were significantly correlated to the level of ventilatory support in PSV, but not in PAV. At the patient's reported comfortable level of ventilatory support, Wi, p, Pdi and PTP decreased by 73%, 74% and 78% respectively in PSV and by 57%, 72% and 65% respectively in PAV. At this level of ventilatory support, improvement of dyspnea was better in PAV than in PSV. Conclusion Both PAV and PSV could relieve dyspnea and load of inspiratory muscles. At the patient's reported comfortable level of ventilatory support, the Wi, p, Pdi and PTP decreased by 57%, 72% and 65% respectively in PAV and by 73%, 74% and 78% respectively in PSV. Patient-ventilator interaction and improvement of dyspnea were better in PAV.

Clinical effects of different exhalation port positions on CO2 elimination during noninvasive positive pressure ventilation (NPPV) were not well studied. Previous lung model studies showed less CO2 rebreathing by placing the exhalation port on the oronasal mask. Patients and methods of study Consecutive patients admitted to the respiratory high-dependency unit were screened for eligibility for study recruitment. Inclusion criteria were: 1) age > 18; 2) hypercapnic respiratory failure with pCO2 > 6 kPa; and 3) in need of ventilatory support for decompensated respiratory failure (pH < 7.35). Exclusion criteria included contraindications to NPPV or refuse to participate in the study. Two exhalation port positions were studied: 1) the Whispering Swivel TM (MWS); or 2) mask exhalation port (MEP) by making a hole on the oronasal mask to serve as an exhalation port. Results At 6 weeks of SARS onset, 25% (30/120) patients had respiratory failure with ALI/ARDS. 16 (53%) had hypercapnia (PaCO 2 > 45 mmHg) during the course of SARS. 28 of them received NPPV therapy. One was intolerable to NPPV treatment. In the remaining 27 patients, NPPV was initiated 1.2 ± 1.6 (0-10) days after onset of respiratory failure. An hour of NPPV therapy led to significant increases in PaO 2 , SpO 2 and PaO 2 /FiO 2 ratio, and decrease in respiratory rate (all P < 0.01). 18 of the 27 patients were weaned successfully from NPPV. The mean duration of NPPV use was 9.9 ± 6.1 (5-30) days. In addition to one patient who was intolerable to NPPV treatment, intubation was required in other 9 patients who initially had a favorable response to NPPV. Remarkable pulmonary barotrauma were noticed in 7 of the 120 (5.8%) patients with SARS. 1 developed days after intubation, 6 occurred while the patients were on NPPV, the incidence was 22% (6/27) in patients using NPPV. The fatality rate at 13 weeks of SARS onset is 6.7% (8/120) in the patient cohort, and 26.7% (8/30) in the ALI/ARDS patients. No SARS related to the care of NPPV patients happened.

Conclusion Some of the SARS patients with ALI/ARDS could experience CO 2 retention, which might be related to the impairment of respiratory muscles. It is proved to be feasible and appropriate for the NPPV treatment of respiratory failure in SARS patients who were at high risk of intubation related complications. As there was a high incidence of remarkable barotrauma, a careful lung protective strategy is necessary during the administration of NPPV as well as invasive mechanical ventilation. 

Non-invasive ventilation is very important in the treatment of acute and chronic respiratory failure and in the respiratory health control. We examined 56 patients with hypoventilation caused by obstructive pulmonary disease and restrictive thoracic diseases. They were admitted for acute respiratory failure with breathing frequency, hypercapnia (Pa > 6.0 kPa), hypoxemia and decreased pH < 7.35. We follow the effects of non-invasive ventilation via standard nose or face mask. The patients were composed of 40 patients with obstructive pulmonary disease (FEV1 = 31, 5% ± 8.0 VC = 36.5% ± 11.0) and 16 patients with restrictive thoracic diseases (FEV1 = 30.5% ± 6.0; VC = 26.6% ± 12.0). Blood gasses were recorded after 2 hours, 1 day, 4 days and 7 days. All patients were receiving oxygen therapy (together with standard treatment for the respiratory disease). We have observed that after 2 hours PaO2 and pH were increased (P < 0.001), after 4 days PaCO2 decreased (P < 0.001). We compared our results with the results of patients without ventilatory support in their home treatment (4 day and 7 day). We have concluded that the support of non-invasive ventilation is important for the patients with respiratory failure and could help in the recovery of their condition.

HENRY KH KWOK 1 , IRIS WS LI 2 , RAYMOND WT LIU 1 , KS LAU 1 , CW LAM 1 1 

Local experience of the use of noninvasive ventilation (NPPV) to assist weaning from invasive mechanical ventilation (MV) was not well studied. Method Records of all intubated patients (from 1 January 2000 to February 2004) admitted to the High Depedence Unit of Ruttonjee and Tang Shiu Kin Hospitals for weaning using noninvasive ventilation as an adjunct were analysed. Results A total of 46 episodes of NPPV weaning during the study period were identified. Four methods of applying NPPV to weaning from MV were identified: (Group A) as a routine systemic method without regards to other weaning criteria (n = 19); (Group B) after the patients failed a spontaneous breathing trial (n = 2); (Group C) when the patient developed postextubation failure after initial successful weaning (n = 13); and (Group D) unplanned extubation (n = 12). More patients with pulmonary fibrosis were treated with routine NPPV after extubation (Group A = 7, Group C = 1, none in Group B and D), but other diagnoses were similar among the four groups. Baseline respiratory rate, astrup and APACHE II scores were also similar. NPPV was applied until the patient was successfully weaned off from all ventilatory support. Group B patients were not further analysed due to its small sample size. The duration of NPPV was significantly more prolonged in Group D patients (mean days ± se) (A: 7.17 ± 1.57, C: 7.40 ± 1.66, D: 14.83 ± 3.08; P = 0.024 for A vs D, P = 0.035 for C vs D). Clinical success rates were significantly higher in Group C patients (A: 57.9%, C: 92.3%, D: 50.0%; P = 0.034 for A vs C, P = 0.018 for C vs D). Hospital mortality rates were as follows: A: 21.1%, B: 50.0%, C 0%, D: 33.3%. A trend towards more prolonged hospital stay (days) in Group D patients was observed (A: 36.44 ± 4.06, C: 25.08 ± 3.72, D: 74.00 ± 23.99; P = 0.057 by one-way ANOVA, P = 0.076 for A vs. D, P = 0.039 for C vs. D).

Conclusions When compared to other groups of patients weaned with NPPV, prolonged weaning with NPPV would be anticipated for patients with unplanned extubation, and these patients would likely need longer hospital length of stay. Also, high success rate could be achieved in patients with postextubation failure treated with NPPV if careful case selection and management were undertaken.

Objective To describe the demographic and physiologic parameters of patients using long term home NIPPV in the form of bi-level positive airway pressure (bipap) device 1 . Method A retrospective descriptive cohort of 27 patients using home bipap during the period from the year 2000 to 2004 was studied. All patients were under the care of a single district general hospital in HK. All underwent manual bipap titration with full PSG in the sleep laboratory for the determination of optimum bipap pressure that achieved optimal oxygen saturation and sleep quality. Their presentations, demographic data, sleep architectures, arterial blood gas during stable phase before and after bipap are described. Mann Whitney Test and Wilconxon Signed Ranks test are used for statistic analysis. Figures are quoted as mean ± 2 SD. Result 13 patients were found to have both obstructive sleep apnea (OSA) and obesity hypoventilation syndrome (OHS). 2 patients had OHS alone. 5 patients had both OSA and chronic obstructive pulmonary disease (COPD). 5 patients had COPD alone. 2 patients had neuromuscular problems 2 . For COPD patients, majority of them presented with type II respiratory failure with frequent repeated admission and difficult weaning. OHS was suspected when obese patients (BMI ജ 25) with hypercapnic respiratory failure could not be explained by other concomitant disease after investigations including HRCT thorax, echocardiogram, lung function test and VQ scan were performed as appropriate. For patients with OHS, the mean age was 58 ± 18 year old, mean body weight was 87 ± 32 kg, mean abdominal girth was 117 ± 19 cm, mean BMI was 34 ± 9 and mean RDI was 63 ± 32/hr. For patients without OHS, mean age was 65 ± 8.7 years, mean body weight was 61 ± 18 kg, mean abdominal girth was 116 ± 15 cm, mean BMI was 26 ± 7.9 and mean RDI was 21 ± 16/hr. BMI, RDI and body weight between two groups were statistically significantly different (P = 0.037, 0.028 and 0.016 respectively). As a whole group, mean REM sleep duration before bipap was 16 ± 24.5 min, after bipap was 46 ± 48.5 min (P = 0.046). CO2 level before bipap was 8.6 ± 2.7 KPa, after bipap was 7.5 ± 1.6 kPa (P = 0.055). Hospital stay before bipap was 13.2 ± 20 days, while after bipap it was shortened to 2.9 ± 5.6 days (P = 0.025). The mean inspiratory positive airway pressure (IPAP) was 18 ± 4 cm water and mean expiratory positive airway pressure (EPAP) was 11.8 ± 5.6 cm water. Conclusion A high proportion of patients using home bipap was found to have features of OHS. They were associated with severe OSA with higher RDI, higher BMI and higher body weight. Apart from COPD, hypercapnic respiratory failure with frequent hospital admissions was found to be a frequent presentation of in-hospital patients with OHS. Therefore further investigation to exclude OHS in this group of patients is warranted. COPD and OHS comprise the major disease category in this bipap cohort 2 . Utilization of bipap improved sleep architecture and arterial blood gas significantly and this has also been shown in another study 3 . Reduction of hospital admissions is also consistent with the finding of other study 4 .

There may be several disadvantages on patient-ventilator interaction in noninvasive mechanical ventilation (NIPPV) whenever much air leak (intentional + unintentional leak) exists. One of them is auto-triggering, which means the ventilator is triggered neither by the patient's inspiratory effort nor by the preset ventilator frequency. This study explored the relationship between air leak and auto-triggering in NIPPV. A quantificational air leak model was set up. The volume of air leak was set as 4 levels, the 1 st level: baseline (only the air leak from the ventilator's expiratory port, 10-30 L/min), the 2 nd level (19-23 L/min air leak adding baseline), the 3 rd level (38-46 L/min air leak adding baseline) and the 4 th level (60-67 L/min air leak adding baseline). Five different types of noninvasive ventilator, a lung model and 14 healthy volunteers (7 males and 7 females) were studied.

The results of lung model study were as follows: 1. Auto-triggering appeared in all of the five ventilators with the increasing level of air leak. 2. Auto-triggering appeared in four ventilators at 1st leak level (only the air leak from the ventilator's expiratory port). The results of volunteers study were: 1. Auto-triggering appeared in all volunteers with the increasing level of air leak. 2. Auto-triggering increased significantly at 2 nd , 3 rd and 4 th leak levels compared with 1 st level. 3. Auto-triggering appeared in four volunteers at 1 st leak level (only the air leak from the ventilator's expiratory port). 4. All the volunteers' respiratory rate increased and tidal volume decreased with the increasing number of auto-triggering. So the rapid shallow breathing index (RSBI) increased. At 2 nd and 3 rd leak level, the mean RSBI was higher than that without NIPPV. At 4 th leak level, the mean RSBI was 2-3 times more than that without NIPPV. 5. The ventilators were triggered before expiratory flow curve returned to base line when auto-triggering appeared. 6. Auto-triggering decreased significantly when less sensitive inspiratory triggering was set, whereas the inspiratory pressure demanded to trigger ventilator increased (3.0 ± 1.4 cmH2O VS 1.2, 1.7 cmH2O, P = 0.000).

In conclusion, auto-triggering occurred when air leak increasing in NIPPV. Autotriggering can impair patients' ventilation and induce PEEPi. The difference between intentional and unintentional is dynamic. Once air leak results in patient-ventilator asynchrony, we should call this volume of air leak unintentional. , and parapneumonic effusions in 2 patients (13.3%). One was diagnosed as having paragonimiasis, from thoracoscopic findings and clinical consideration. There was no procedure-associated mortality. There were six cases of new onset fever (40%) and one case of pneumothorax (6.7%). Two millimeter minithoracoscopy which is less invasive than conventional thoracoscopy was an accurate and safe method for undiagnosed exudative pleural effusion. Carcinoma of the lung was the commonest cause of malignant effusions and bronchoscopic biopsy gave the highest yield of histological diagnosis (66%), followed by pleural fluid cytology (59%) and pleural biopsy (50%). The combination of these three procedures increased the diagnostic yield to 96%. In tuberculous pleural effusion, pleural fluid staining for acid-fast bacilli staining was negative in all cases but mycobacterial culture was positive in 24% of cases while pleural biopsy gave a better yield of 68%. Examination of sputum and bronchoalveolar lavage specimens confirmed the diagnosis of tuberculosis in 40% of cases. A combination of these investigations yielded the diagnosis in 92% of patients with tuberculous effusion. Conclusions Pleural fluid analysis, pleural biopsy and fibreoptic bronchoscopy are helpful in diagnosing tuberculous and neoplastic effusions due to carcinoma of the lung. We describe a case of a 39 year old male who was admitted in our hospital due to persistent bleeding from the biopsy site done on the right breast mass. Our patient presented with tuberculosis-like symptomatology, a breast mass mimicking a hemangioma and on chest CT scan, an empyema-like density with extrathoracic extension. He underwent open thoracotomy and chest wall reconstruction. Histopathologic examination revealed epithelioid angiosarcoma. The tumor cells showed positive staining for CD34 and Factor VIII and were negative for smooth muscle actin. Our patient is currently on his 6th month post surgery. Though functionally well, the note of tumor recurrence and presence of adenopathies signal a bad prognosis. To our knowledge, this is the first case of its kind reported in our country. Primary thoracic epithelioid angiosarcoma is an extremely rare neoplasm with only a few cases reported thus far in the international literature. This highly aggressive tumor, which may arise in various sites, is part of the continuum of the family of epithelioid vascular tumors. Recurrence and behavior of this tumor is unpredictable. It carries a dismal prognosis. No single effective therapy has yet been established, though a combination of surgery, radiotherapy and chemotherapy, as well as immunotherapy have been tried with variable results. Although uncommon, primary intrathoracic angiosarcoma must always remain in the differential diagnosis of pleural and subpleural masses in the presence of pulmonary ascites, nodules, diffuse infiltrates and an opacified lung.

Objectives To identify the expression of AQP 1 in visceral and parietal pleura in SD rat and to examine the effect of AQP 1 in pleural fluid turnover. Methods Pleural membranes were carefully microdissected from normal SD rats. Immunohistochemistry was done using specific goat polyclonal antibodies. SD rats were divided into five groups, including control group (normal saline, n = 8), dexamethasone group (n = 24), erythromycin group (n = 24), hyper osmotic NaCl group (n = 24), and lipopolysaccharide group (n = 24). Agents were intrapleural injected. Pleural tissues were homogenized in Trizol for mRNA isolation. Real time RT-PCR was carried out with AQP 1 gene-specific primers. Results AQP 1 was immunolocalized predominantly to the microvessels and mesothelial cells of visceral and parietal pleura. The extent of AQP 1 expression in parietal pleura was less than that in visceral pleura. AQP 1 expression increased at all phases in dexamethasone group and hyper osmotic NaCl group, whereas it decreased in erythromycin group and lipopolysaccharide group. Conclusion Enhanced expression of AQP 1 may relate to more absorption in pleural effusion by dexamethasone. Rapid osmotic equilibration across the pleural surface is facilitated by AQP 1 . Both erythromycin and lipopolysaccharide downregulated AQP 1 expression suggests a possible mechanism for AQP 1 modulation by inflammatory cytokines. Our experiment provides indirect evidence that AQP 1 may play a role in the pleural fluid turnover. However, animal model of pleural effusion under conditions of stress (i.e. TB, bacteria, virus etc), AQP 1 gene knockout animal and inhibitor of AQP 1 are needed to confirm the role of AQP 1 in pleural fluid transport. Polymerase chain reaction Polymerase chain reaction (PCR) is a common method of creating copies of specific fragments of DNA. PCR rapidly amplifies a single DNA molecule into many billions of molecules and can be detected by visualization. Setting Diagnostic of pleural effusion caused by Mycobacterium tuberculosis is still a clinical problem especially of low sensitivity in conventional methods. Objective To evaluate PCR technique based on IS6110 sequence detection of M. tuberculosis in pleural fluid. Material and method Subjects: A total of 62 pleural fluid samples of highly suspected pleurisy tuberculosis outpatient and inpatient of Persahabatan Hospital, Jakarta. Diagnosis made based on microscopically, microbiological and histopathological finding. Each sample was examined microscopic examination with auramin and Ziehl-Neelsen stain, culture in solid and liquid media, pleural biopsy for histopathology examination, PCR with E1 and E2 primer. Gold standard criteria for pleurisy tuberculosis were positive if one or more examination positive (microscopic, culture, histopathology). DNA was extracted with Boom method. Primer E1 and E2 were used and amplified to 123 bp of IS6110. Sample was exclude if the culture grew mycobacteria other than tuberculosis. Results PCR was a rapid diagnostic test and can detect DNA of M. tuberculosis in pleural fluid specimen with sensitivity 54.35%, specificity 93.75%, positive predicted value 96.15%, negative predicted value 41.67%, false positive 1 (1.61%), false negative 21 (33.87%) (c 2 = 11,278, P = 0.001). Sensitivity of direct stain AFB, culture and pleural biopsy were 12.77%, 23.40%, 96.55% and the specificity were 100%, 100%, 45.45% respectively. Quality control of laboratory, primer used, contaminant and dead bacilli were influenced factors of PCR result. Conclusions PCR has higher diagnostic value of pleurisy tuberculosis compared with AFB microscopy, culture but lower than pleural biopsy, however it should be carefully implemented with patient's clinical condition. Keyword: pleuritis tuberculosis, polymerase chain reaction, diagnostic Background The intrapleural injection of transforming growth factor-beta2 (TGF-b 2 ) or doxycycline produces excellent pleurodesis in rabbit model. However, the intrapleural injection of these agents induces large pleural effusion which is possibly related to vascular endothelial growth factor (VEGF). The aim of this study was to investigate whether anti-VEGF antibody has any effect on the fluid production or the pleurodesis induced by TGF-b 2 or doxycycline in rabbits. Methods Two groups of 8 New Zealand white rabbits were given doxycycline 10 mg/kg intrapleurally after chest tube placement. The doxycycline treatment group received 10 mg/kg anti-VEGF antibody intravenously 24 h before doxycycline injection and the doxycycline control group received no Anti-VEGF antibody. Another two groups of 8 New Zealand white rabbits were given TGF-b 2 5.0 mg intrapleurally. The TGF-b 2 treatment group received anti-VEGF antibody 10 mg/kg intravenously 24 hours before TGF-b 2 injection and the TGF-b 2 control group received no antibody. The rabbits were sacrificed at 2 weeks and the pleurodesis score was graded macroscopically on a 1-8 scale. The degree of angiogenesis in pleural tissues was assessed by immunohistochemical staining for factor VIII which was assessed by computer-assisted digital analysis.

The administration of anti-VEGF antibodies had no effect on pleural fluid volume or the characteristics of the fluid. The mean pleurodesis score of TGF-b 2 control group (7.71 ± 0.76) was significantly (P < 0.05) higher than antibody pre-treatment TGF-b 2 group (4.43 ± 2.37). The mean pleurodesis scores of doxycycline control group (6.0 ± 1.69) was significantly (P < 0.05) higher than that for antibody pre-treatment doxycycline group (2.0 ± 0.93). The administration of the anti-VEGF antibody also reduced the angiogenesis. The percent of pleural tissue demonstrating angiogenesis in the TGF-b 2 control group (4.87 ± 0.43%) was significantly (P < 0.05) higher than that for antibody treatment TGF-b 2 group (2.94 ± 0.68%). The percent of pleural angiogenesis in the doxycycline control group (6.86 ± 2.20%) was significantly (P < 0.05) higher than the antibody pretreatment doxycycline group (2.17 ± 0.90%).

Anti-VEGF antibody significantly inhibits the pleurodesis induced by doxycycline or TGF-b 2 . This observation suggests that VEGF and angiogenesis play a pivotal role in the production of a pleurodesis. Pleurodesis may be not effective in pleural effusion patients treated with novel anti-angiogenic regimens that block VEGF. 

Objective To detect the level and dynamic changes of IgG, IgM antibodies against SARS-CoV in the one year recoveried SARS patients, the aim is to provide some information for this virus's prevention and vaccine's research. Methods The IgG, IgM antibodies against coronavirus was detected with ELISA in the blood of convalescent SARS patients for every 2-4 weeks. To the convalescens that either IgG or IgM antibody continuously positive with twice will be selected to going on detection, while to that of both IgG and IgM antibody continuously negative with twice will be excluded. Stata 7.0 statistics software was used to analysis the results. Background Perforin is a 66-kDa protein, contained in granules of natural killer (NK), NKT, gamma-delta T, and killer T cells. Besides to tumor immunity, it contributes to host protection against infection by virus and intracellular bacteria such as Mycobacterium tuberculosis through inducing apoptosis in infected cells. In this study, we investigated the role of perforin in bacterial infection, especially focusing on extracellular pathogen, Streptococcus pneumoniae. Methods Perforin-deficient mice (perforin-/-), C57Bl/6 background, were used in this study. We used two clinical isolates of S. pneumoniae. One was a virulent strain, capsular-serotype 3 and another was a low virulent strain, capsular-serotype 6. The organisms were administered intratracheally or intraperitoneally. Mortality rate of mice and proliferation of bacteria in the lungs or peritoneal cavity were investigated. Results After administration of serotype 3 S. pneumoniae, both wild and perforin-/-died within three days. There was no difference in mortality rate.

In the case of serotype 6 S. pneumoniae, mortality of perforin-/-was significantly higher compared to wild. Proliferation of S. pneumoniae was also increased. There was no difference in bacterial phagocytosis by macrophages in vitro. Since NKT cell has been reported to have a critical role in S. pneumoniae infection, alpha-galactosylceramide was coadministered. It clearly improved mortality of mice and inhibited proliferation of S. pneumoniae. Conclusion This study indicated that perforin has a certain role in innate immunity against extracellular bacteria infection via NKT cell activation.

Further study should be focused on the roles of perforin and NKT cells in S. pneumonia infection. Results Pulmonary vascular permeability and wet/dry ratio in immunocompromised group were higher than that in control group. Pulmonary congestion and hemorrhage were more severe in immunocompromised group compared with the control group. The iNOS mRNA expression in immunocompromised group was much higher than that in control group (0.71 ± 0.09 vs 0.51 ± 0.06, 9 hs), however, the eNOS mRNA expression was lower in immunocompromised group than that in control group (0.31 ± 0.04 vs 0.42 ± 0.03, 9 hs). There has marked correlation between iNOS and items of vascular injury, and eNOS too. The survival rate of in immunocompromised group was lower than that in control group (79.2% vs 97.6%). Conclusion The overexpressive iNOS level and relative lower eNOS level may contributed to the severe lung vascular injury and the lower survival rate in pneumonia of immunocompromised rats. Selectively anti-iNOS antibodies may be helpful to alleviate the lung vascular injury in immunocompromised rats' pneumonia. ; P = 0.001) and total white blood cell count of 12 ¥ 10 9 /L or less on admission (OR 3.41; 95% CI, 1.05-11.08; P = 0.041).

Conclusions Mycobacterium tuberculosis is a common cause of CAP requiring hospitalization in Malaysia. Patients with PTB had a longer duration of symptoms, more frequent upper lobe involvement and lower total white blood cell count than patients with non-TB CAP.

Background As modern medicine developed, the quantity of granulocytopenia became larger than ever. Patients with granulocytopenia are apt to develop severe pneumonia and acute lung injury. Previous study show that there has an unbalance of oxidant-antioxidant system in pneumonia of granulocytopenia hosts. To evaluate the role of antioxidant treatment in pneumonia of granulocytopenia immunocompromised hosts, we designed this research. Methods Sprague-Dawley rats were used to establish model of immunocompromised rats' pneumonia. Using N-acetyl-L-cysteine as antioxidant to treat granulocytopenia rats. Observed a time-course of following items such as the amount of peripheral blood cell, rats' mortality, bacterial loading of homogenized lung tissue, superoxide dismutase and malondialdehyde of rat's serum and lung, the influence of pulmonary vascular permeability and the shift of lung edema, changes of pulmonary histopathology, etc.

Results Treated group has a lower mortality than control group (16.3% vs. 23.4%). Superoxide dismutase of the treated rats were higher than control group in both serum and lung tissue, while malondialdehyde were lower than control group. Pulmonary vascular permeability and wet/dry ratio of treated group were moderate than control. At 9 hours after infection, the protein ratio of BALF and serum was 0.03 ± 0.005 in treated group, while it was 0.05 ± 0.007 in control group. And the wet/dry ratios of two group were 8.2 ± 0.9 vs. 10.8 ± 1.2 at 9 hours after infection. Pulmonary histopathology of treated group represent moderate lung injury, pulmonary congestion and hemorrhage than control group. Conclusion Antioxidant treatment in pneumonia of granulocytopenia rats is proved helpful in alleviating acute lung injury and decreasing mortality. It may be a important accessorial means in treating granulocytopenia pneumonia.

Objective To determine the clinical features associated with mortality in patients with community acquired pneumonia (CAP) requiring hospitalisation. Backgroud To establish a set of clinical comprehensive score system to evaluate the condition and prognosis of the patients with severe acute respiratory syndrome (SARS). Methods We establish a set of clinical comprehensive score system based on the patients' symptom, such as fever and shortness of breath, chest radiography and computerized tomography (CT) scan changes, arterial blood oxygen, serum album level and subsets of blood T lymphocyte. Total score is 34. 46 cases of SARS respectively from Shanghai Pulmonary Hospital and Shenzhen Donghu Hospital were retrospectively evaluated by this system. Results Each separate score of above items had a positive correlation with the clinical comprehensive score, the correlation coefficients (g) are from 0.481 to 0.897, P value are all less than 0.01. There were 13% of patients (6 of 46 patients) got score less than or equal to 9, their conditions were all slight, recovered and discharged in the end. There were 65.3% of them (30 of 46 patients) got score more than or equal to 10 and less than 20, their conditions were secondary severe, 83.0% of them (25 of 30 patients) restored entirely and discharged, 17% (5 of 30) either were transferred to other hospital for further treatment (2 of 5) or had some extent lung fibrosis (3 of 5). There were 21.7% of patients (10 of 46) got score more than or equal to 20, their conditions were very severe, 50% of severe patients (5 of 10) recovered entirely and discharged, 30% of them (3 of 10 patients) had lung fibrosis, 20% died (2 of 10 patients). The patients with lung fibrosis were all got score equal or over 10. Died patients' conditions were all very severe and they got highest clinical comprehensive score, each of separated items score they got was almost highest. These results suggested that the more clinical comprehensive score that the patients got, the more severe of their conditions and the worse prognosis of them. Conclusion This set of clinical comprehensive score system can reflect the severe extent of SARS patients' condition quite objectively and can be used for evaluating SARS patients' condition and estimating their prognosis.

LA ROSENTHAL 1 , SP AMINEVA 2 , RJ SZAKALY 1 , JE GERN 2 , RF LEMANSKE 1,2 , RL SORKNESS 1,2,3

Background Rhinoviruses, members of the picornavirus family, induce neutrophil-rich inflammatory responses and are major causes of asthma exacerbations, which may involve lower airway infections. There are no known rodent-specific rhinoviruses, but rodents are natural hosts of the picornavirus, mengovirus, an inducer of encephalitis and myocarditis that has not been shown to cause respiratory infections. We hypothesized that inhalation of an attenuated mengovirus, vMC 0 , would induce a respiratory infection in rats that could be a useful model of human rhinovirus infections. Methods BN/SsN rats were inoculated with vMC 0 , UV-inactivated vMC 0 , or vehicle intranasally or by lung insufflation. On days 1, 3, or 5 postinoculation, viral titers in lung homogenates and bronchoalveolar lavage (BAL) fluids were measured by plaque assay; differential BAL cell numbers were determined; and giemsa-stained lung sections were examined for cellular infiltrates.

Results Inoculation of rats with vMC 0 , but not vehicle or UV-inactivated vMC 0 , resulted in the presence of active virus in lung homogenates and BAL fluid on days 1 and 3, but not 5, postinoculation and marked pulmonary inflammation, peaking on day 3 postinoculation, as shown by significant 159-, 28-, 7-, and 2-fold increases (P < 0.04) in median numbers of BAL neutrophils, lymphocytes, eosinophils, and macrophages, respectively, and by extensive cellular infiltrates in lung sections. Virus-related signs/symptoms were not observed in non-respiratory organs. Background Community-acquired pneumonia (CAP) is most often a bacterial disease with a substantial mortality. The antibiotics prescribe decision should be based on precise diagnosis, assessment of the severity of the disease and epidemiological information.

Objective We did the survey nationwide to analysis the spectrum of microbiological agents causing CAP in recent years. We also investigated the procedure of diagnosis as well as the empirical treatment for this disease in OPD (outpatient department) of pulmonary disease in China. ) was the commonest pathogens followed by Pseudomonas (4%). Quinolones (47%) were most frequently prescribed followed by 3 rd Generation Cephalosporin (41%). Quinolones remained the most commonly prescribed antibiotics even after antibiotics were changed during hospital discharge and after the result of culture data (60.7%).

Conclusions The empirical antibiotics used are different from those recommended in international guidelines. Concerns about penicillin resistant were reflected in frequent use of quinolones and cephalosporin.

Pyocyanin, a blue phenazine pigment produced by Pseudomonas aeruginosa, has been shown to stimulate the release of interleukin (IL)-8 in airway epithelium directly, as well as synergising with IL-1 and tumour necrosis factor (TNF) in triggering IL-8 release. IL-8 is a potent chemoattractant that accounts for the neutrophilic influx seen in the pathogenesis of bronchiectasis. Mechanisms regulating IL-8 expression have been shown to involve protein kinase C (PKC) activation. In this study we investigated the effects of pyocyanin, alone and in combination, with IL-1b and phorbol 12,13-dibutyrate (PDBu) on IL-8 mRNA and release in the human bronchial epithelial cell line (BEAS-2B) by reverse transcriptasepolymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). RT-PCR and ELISA showed that pyocyanin alone produced no increase in IL-8 mRNA or IL-8 release. However, pyocyanin caused significant potentiation in IL-8 release triggered by IL-1b and PDBu in a dose-dependent manner without affecting IL-8 mRNA levels. Incubation with dexamethasone, budesonide, fluticasone, and erythromycin caused dosedependent suppression of IL-8 release stimulated by pyocyanin plus IL-1b or PDBu. Budesonide and fluticasone were 10-fold more potent than dexamethasone in the inhibition whilst erythromycin exerted its effect at a much higher dose starting at 0.3 mM. Our findings suggest that potentiation of cytokine-or PDBu-induced IL-8 release by pyocyanin may be due to post-transcriptional mechanisms. These data confirm the established role of glucocorticosteroids, and erythromycin as anti-inflammatory action in treating bronchiectasis. Objective To investigate the in vivo efficacies of ceftazidime and cefepime in rats with experimental pneumonia due to extended-spectrum b-lactamaseproducing strain of Klebsiella pneumonia sus ceptible to either of the antibiotics in vitro. Methods With intratracheal method, 3 Klebsiella pneumonia strains isolated from this hospital were used to produce 3 groups of the experimental model of pneumonia in rats. All the 3 strains showed resistant to cefotaxime and susceptible to piperacillin-tazobactam in vitro. To ceftazidime and cefepime, strain 1 were both susceptible. Strain 2 were susceptible to ceftazidime and resistant to cefepime. Strain 3 were resistant to ceftazidime and susceptible to cefepime. The three groups of rats were randomly assigned to one of the following five groups: one control group and four treatment group of the four above antibiotics. 96 hours later, the efficacies were evaluated by the survival rate and the viable bacterial counts of lungs (lgCFU/g). 

Objective To investigate the clinical characteristics in the fastigium stage of severe acute respiratory syndrome (SARS) patients with positive specific anti-SARS coronavirus antibody. Methods Retrospectively analyzed the data of 122 SARS patients whose serum anti-SARS coronavirus IgG antibody were positive (positive group) contrasted by 50 SARS patients with negative specific antibody (negative group) to analyze the clinical characteristics. In the positive group, contrasted the common case with the critical case to analyze the major influencing factors.

The history of close epidemiological contact, the initial symptom of pyrexia, (fastigium stage) the body temperature ജ38°C or ജ39°C, white mucous expectoration, rapid progress in chest radiographs, PaO 2 < 60 mmHg, increase in AST, increase in LDH, increase in CK (P < 0.01) and chest distress/short breath, decrease in lymphocytes (<0.9 ¥ 10 9 /L), white lung shallow in chest radiographs (P < 0.05) were seen often in the positive group. Yellow purulent expectoration was seen often in the negative group (P < 0.01). Chest distress/short breath, white mucous expectoration, PaO 2 < 60 mmHg/PaO 2 < 80 mmHg, increase in LDH, rapid progress or white lung/bilateral/multiple/diffuse shallow in chest radiographs (P < 0.01 or P < 0.05) were seen often in the critical case.

Conclusions The history of close epidemiological contact, the initial symptom of pyrexia, (fastigium stage) the body temperature ജ38°C, chest distress/short breath, white mucous expectoration, rapid progress or white lung shallow in chest radiographs, PaO 2 < 60 mmHg, decrease in lymphocytes (<0.9 ¥ 10 9 /L), increase in LDH were the major factors on the differential diagnosis and critical outcome in the fastigium stage of SARS 

Objective To know about the clinical features, the changes in radiography, the pathological findings of pulmonary cryptococcosis (PC) and its diagnosis and treatment through cases analysis. Method 23 cases with PC in our hospital since 1980 were studied. Their symptoms, chest radiograph manifestation, pathological and therapeutic findings were analyzed.

Result (1) The ratio of male to female in these cases was 18 to 5 with mean age of 42.17 ± 7.34 years old and the most (87.0%) were from 28 to 55 years old. Only two cases (8.7%) had underlying diseases.

(2) Their symptoms were mild and non-specific, such as mild cough, fever, night sweat. Some cases were found occasionally in physical examination (PE).

(3) The chest radiograph manifestation were various. Well-defined solitary masses with 3-10 cm in diameter, were found solitary in 14 cases (60.9%); single or multiple large patchy high density with small cavity in 5 cases (21.7%); single nodule in 2 cases (8.7%) while multiple nodules in 1 case (4.3%) and solitary nodule with pleural effusion in 1 case (4.3%). Most of abnormality located in surrounding pleurae. (4) All cases were diagnosed by pathologic examinations and all proved to be cryptococcus granulomas. 17 cases were correctly diagnosed through surgical section, 3 through VATS lung biopsy, 2 by CT-guided transthoracic needle aspiration biopsy and 1 by transbronchoscopic lung biopsy (TBLB). (5) All cases received treatment. Among them, surgical section were performed in 17 cases, and 9 cases of them accepted post-operation antifungal drugs. They all recovered well expect one case, who died of simultaneously occurred cryptococcal meningitis soon after surgery. Only 7 of cases could get follow-up, the longest was 15 years, and no reoccurrence was found. 6 cases were treated only by antifungal drugs. The duration of follow-up was from 2 months to more than 3 years. Shadows in chest radiograph in 4 cases vanished basically, 1 case decreased apparently in size while it became smaller a little in 1 case. Conclusion It should be alert to PC when the following happen: middle age patient, especially male; mild cough, fever, night sweat; surrounding well-defined solitary mass, large patchy high density with small cavity in chest radiography manifestation. And we should undertake the examination of pathogen and pathology at the beginning. The key to improve the curative rate was early diagnosed and rational therapy. Objective To analyze the clinical diagnostic criteria for serious SARS in Guangdong province retrospectively and discriminately with SARS database. And to screen out the sensitive warning factor. Methods 402 SARS patients were selected based on the diagnostic criteria for SARS from ministry of health, P. R. China. And 358 SARS patients were selected again according to serious diagnostic criteria. The study subjects were divided into 2 groups. One group was the specific serious SARS among in 358 patients which were accepted invasive or non-invasive mechanical ventilation or die of the disease. Remain 44 SARS patients as the non-serious SARS group. We using the oxygen index (OI) diagnosis such as acute lung injury (ALI) or acute respiratory distress syndrome to evaluate the serious SARS.

Results OI less than 300 were identify the specific serious SARS patients, the mistake judge rate less 0.0062. And further more we compared two group's mordality and incorporate syndrome with Logistic regression which kick out the overlap SARS patients. The results showed significant difference in two groups. Conclusion Oxygen index less than 300 identify for ALI could meet the qualification criteria of serious SARS. And may become a warning factor in clinical use. Background Invasive pulmonary aspergillosis (IPA) is a common and devastating complication of immunosuppression. Dendritic cells (DCs) behave as both sentinel for innate immune recognition and initiator for protective immunity. IL-12 can induce IFN-g production and enhance proliferation of Ag-specific T cells.

Objective To study the role of DCs in the control of aspergillus infection and assess a new paradigm in the development of a vaccine to protect against the infection. Methods Engineered to secrete IL-12 and pulsed with aspergillus, bone marrowderived DCs were administered via the lateral tail vein to syngeneic mice. The immunosuppressed mice model was made through intraperitoneal administeration of cyclophosphamide and then a suspension of aspergillus conidia was applied to the nostrils to obtained the IPA model. The electron microscopy and flow cytometry analysis were analyzed and the bacteriology, pathology, cytokines in the lung of mice with IPA were valued. Results DCs can engulf aspergillus conidia through coiling phagocytosis and then mature. Transfection of DCs with adenovirus did not affect their capacity to engulf conidia. Adoptive transfer of DCs pulsed ex vivo with heat-inactivated Af (HAF) to naive mice induced the Ag-specific production of IFN-g, and transfection of these DCs augmented this immune response. Animals receiving HAF-pulsed DCs had lower fungal burdens, a higher survival rate. IL-12-engineered DCs could augment this protection strikingly as judged by a higher survival and that almost no aspergillus could be detected in the lung of mice that had received IL-12transduced, HAF-pulsed DCs. Conclusion DCs play major roles in host defense against aspergillus fumigatus. Our findings offer encouragement and additional rationale for the development of a DC-based vaccine against aspergillus and perhaps other infectious diseases.

[This work was supported by the Shanghai Health Care Systems (Bai Ren Project, Grant 98BR030).] Patiensts with chronic respiratory disease, such as chronic bronchitis, obstructive emphysema, bronchial asthma, bronchiectasis or chronic cardiac disease, and with symptoms of influenza were enrolled. They should satisfy the following criteria: Fever ≥37.8°C plus at least two of the following influenza symptoms: coryza/nasal congestion, sore throat, cough, myalgia/muscles aches and pain, fatigue, headache or chills/sweats. And the post onset of feeling unwell should be within 48 hours. Individuals were randomly signed in Oseltamivir group (oseltamivir 75 mg, twice daily for 5 days) or control group who were given symptom relief medicine.

Results 56 of the 108 recruited individuals were identified as influenza-infected through laboratory test. They were defined as intent-to-treat infected population (ITTI) (27 Oseltamivir, 29 control). The duration of influenza symptom was 64 hours shortened (36.7%) and AUC score of the influenza symptom was decreased by 618 (43.1%) in oseltamivir group compared with that in the control group. For the fever lasting duration, it is 46.8 hours (45.0%) less in the Oseltamivir group than that in the control group. It took 6 days in the Oseltamivir group and 11 days in the control group to come back to the basic health status. Secondary complications such as bronchitis, sinusitis and pneumonia occurred 11% (3/27) in Oseltamivir group and 45% (13/29) in the control group. The treatment expense of influenza and its complication were 587.4 RMB in the Oseltamivir group and 786.5 RMB in the control group, which has no significant difference (P = 0.2462). Conclusion Our data suggested that Oseltamivir is effective and well tolerated in the chronic respiratory and cardiac disease. It can reduce the fever duration and severity of influenza symptom, decrease the incidence of secondary complications, antibiotic using, and does not increase the total medical cost.

Objective To study rat's pulmonary inflammatory reaction induced by N-protein of SARS-coV and effects of glucocorticoids. Methods Rat's pulmonary inflammatory reaction were induced by intratracheally instillation of N-protein of SARS-coV with a dose of 0.2 mg/kg. Rats were randomly divided into four groups: saline controls (NC), N-protein group one (P1, 6 h), N-protein group two (P2, 24 h), dexamethasone group (D, 10 mg/kg). Their blood, bronchial alveolar lavage fluid (BALF) and lung tissue were collected after challenging. Cytological and histopathologic Changes of lung tissue were observed. The wet/dry ratio (W/D) of lung tissue were determined. Interleukin-6 (IL-6), interleukin-10 (IL-10), transforming growth faCtor-b1(TGF-b1) of serum and BALF were measured by ELISA.

Results ( (4) Compared with NC group, IL-6, IL-10, TGF-b1 levels in both serum and BALF of P1 group increased significantly (P < 0.05), these cytokines levels in both serum and BALF of P2 group were higher than P1 group (P < 0.01); these cytokines levels in D group decreased noticeably vs P2 group. Conclusions: N-protein of SARS-coV can lead to rat's pulmonary inflammatory reaction/acute lung injury which related to the rise and unbalance of proinflammatory and anti-inflammatory cytokines; glucocorticoids can effectively alleviate pulmonary inflammatory reaction induced by N-protein of SARS-coV. Objective To explore the relationship between arousal and nocturnal heart rate variability (HRV) in patients with obstructive sleep apnea hypopnea syndrome. Methods According to overnight polysomnography (PSG), 27 patients with moderate to severe OSAHS were enrolled, except cardiopulmonary disorders or nervous system diseases. For all patients, choose an hour continuous PSG record in non-rapid eye movement (NREM). Compare the mean HR within 10 seconds before onset of each arousal with the peak HR within 10 seconds after onset of arousal during this hour. At the same time, calculate B-ArI and pulse rate rise index (PRRI), then make correlation analysis. Furthermore, compare the DHR at apnea termination between 10 events with and without EEG arousal (matched according to minSaO 2 ) in NREM in 18 patients with moderate to severe OSAHS.

The peak HR within 10 seconds after onset of arousal was higher compared with the mean HR within 10 seconds before onset of arousal, B-ArI was positively related to PRRI (r = 0.97, P < 0.001). The DHR of those events with EEG arousal at apnea termination was higher than those without EEG arousal. Background Obstructive Sleep Apnea (OSA) is a condition characterized by repeated episodes of apnea and hypopnea. There has been increasing recognition of OSA with increasing prevalence estimates. It has brought about several clinical consequences most of which are preventable upon treatment of OSA. Given the prevalence of OSA in the population and its dire outcomes, accurate screening for positive cases is being done. The aim of this study is to assess the accuracy of nocturnal pulse oximetry as a confirmatory test in the diagnosis of OSA as against polysomnography (PSG) which is the gold standard. Design Cross-sectional Study. Setting St. Luke's Medical Center Comprehensive Sleep Disorders Center.

Center for PSG with symptoms suggestive of OSA were investigated. Full night diagnostic polysomnography with oximetry was performed at the sleep center. Baseline characteristics were taken. Respiratory Disturbance Index (RDI) and Lowest O2 Saturation (LSAT) levels were determined for each patient. Five LSAT threshold values (£75%, £80%, £85%, £90% and £95%) were obtained for analysis. Sensitivity, specificity, false positive rate, positive predictive values (PPV) and negative predictive values were computed for each threshold level. Receiver operating characteristic (ROC) curve was constructed to measure the diagnostic performance of LSAT.

Results 416 were included in the present study after meeting the inclusion criteria. The group consisted of 315 (75%) men and 101 (25%) women. Mean age ± SD was 46.02 ± 12.07. The average BMI ± SD was 29.04 ± 6.10. The mean RDI was 33.27 ± 36.37. 284 patients had an RDI of ≥5/hour. Of these 244 (86%) were men and 40 (14%) were women. When compared statistically, there was a significant difference in the BMI and LSAT. Correlation of RDI ≥5/hour with LSAT was statistically significant at P = 0.000 with an r = -0.662. Using LSAT threshold of £85% gave a sensitivity of 83%, specificity of 90% with FPR of 10%, PPV of 95% and NPV of 72%. AUC = 0.93. Conclusion LSAT level of £85% may suggest that patients with symptoms referrable to OSAS has OSA. For adults who are unlikely to have OSA clinically, a negative oximetry result may be sufficient to exclude the diagnosis; for adults who are likely to have OSA clinically, a positive oximetry result may be sufficient to confirm the diagnosis of OSA.

Background and Objectives The magnitude of the problem of sleep-related breathing disorders and the gender differences of these disorders are not well studied in India. Therefore a study was undertaken in the rural population of Delhi to know the gender differences in the prevalence of sleep-related breathing symptoms.

Methods Nineteen out of 232 villages in Delhi were randomly selected. Households were then selected randomly to obtain a sample of 350-400 subjects of either sex from each village. The study was done using a questionnaire. The field investigators made house-to-house visits and administered the questionnaire to all over 18 years of age. A repeat visit was made by the Investigators to contact the members absent during the first visit. The questionnaire had fifteen multiplechoice questions and each question was scored depending on the severity of symptoms. Each subject has to choose one of five possible alternatives for each question: 'never', 'less than once a week', 'once or twice a week', three to five nights/days a week', or 'almost every day/night'. The respondents were classified as having sleep-related breathing symptoms if they had loud snoring (scores 4 or 5) and/or daytime sleepiness (scores 4 or 5). Age, weight and height were recorded.

Results A total of 6908 subjects from the rural area of Delhi were studied. There were 3562 (51%) males and 3346 (49%) females. A history of seep-related breathing symptoms was present in 448 (12.5%) males and 315 (9%) females. Snoring was significantly more in males (P < 0.001). However, daytime sleepiness (both during working and leisure activities), morning headache, memory loss, tiredness, nightmares and nocturnal awakening (P < 0.001, for each comparison) were higher in females. Sleep-related breathing symptoms increased as age advanced in both sexes. Subjects with sleep-related symptoms were older (P < 0.001) and had weight (P < 0.001) and body mass index (P < 0.001) higher in both sexes compared to subjects without such symptoms. Conclusion The prevalence of sleep-related breathing disorders was 12.5% in males and 9% in females of Delhi rural population. Snoring was significantly higher in male subjects. Other sleep related-breathing disorders (daytime sleepiness, morning headache, memory loss, tiredness, nightmares and nocturnal awakening) were higher in females. (Supported by the Department of Science and Technology, Government of India.)

Introduction Central sleep apnea (CSA) is a disorder with a variable clinical presentation. Polysomnographically, CSA is characterized by recurrent episodes of absent/decreased respiratory effort during sleep. Abnormalities in gain of metabolic ventilatory control system or circulatory abnormalities may contribute to unstable, periodic ventilation during sleep. Acetazolamoide (ACET) is known to potentiate the ventilatory response to chemical stimuli and to alter brain blood flow. Likely, via one or both these mechanisms, ACET has been found to improve sleep-induced periodic breathing and central sleep apnea. Worsening of obstructive sleep apnea (OSA) has also been reported with ACET. In addition, previous studies indicate that a heightened ventilatory drive may contribute to OSA. Based on these concepts, we hypothesized that in patients, primarily with CSA but with mild OSA, CSA would be improved and OSA worsened by ACET. Methods Patients were selected with a Central AH index ≥5/h and OSA index £5/hr. Central Apnea/Hypopnea was defined as a >80%/>50% reduction in oral/nasal flow, accompanied by a proportional thoracic and abdominal decrease in effort measured by thoracic and abdominal strain gauges. OSA was defined as obstructive AH index ≥5/h and central AH index £5/h. Obstructive apnea/hypopnea was defined as reduction in oral/nasal flow, accompanied by persistence or an increase in thoracic and abdominal effort. In an open label, prospective study of 8 patients diagnosed with CSA, ACET 250 mg orally bid was administered for 6 weeks, followed by repeat polysomnogram (PSG) in the last week of therapy.

Results Average age was sixty-one, race distribution C/AA 4/4 and gender difference M/F 7/1. Respiratory disturbance index (RDI) decreased by approximately 50%, but because of variability and small group size, this was not statistically significant (42 ± 12 vs. 20 ± 14 P = NS). Central apnea index (16 ± 16 vs. 2 ± 3 P = 0.0001) and Central hypopnea index (20 ± 15 vs. 3 ± 4 P = 0.019) improved during NREM and REM sleep. Obstructive AH index (1 ± 2 vs. 15 ± 10 P = 0.001) worsened with ACET. ACET use did not improve Epworth sleepiness scale (ESS) (12 ± 5 vs. 9 ± 5 P = NS) Conclusion Acetazolamide tended to decrease the overall RDI, but primarily changed the sleep disordered breathing pattern in CSA patients. Mild excessive daytime sleepiness persisted with ACET. We speculate that ACET may influence respiratory pattern generation but worsen factors contributing to upper airway collapse during sleep. Objectives To evaluate the short and long-term changes of the level of continuous positive airway pressure (CPAP) in patients with obstructive sleep apnea hypopnea syndrome (OSAHS), and address the necessity of re-titration of CPAP pressure during long term treatment.

Methods Twenty-five patients diagnosed as moderate to severe OSAHS (AHI >15 with a mean of 56) were included to the study. A manual overnight titration to determine the optimal CPAP pressure (oCPAP) was undertaken following the diagnostic PSG test. Then all of the 25 patients were admitted to sleep ward for a one week auto-CPAP treatment (418A with software, Tyco Health Care), SpO2 and PSG were detected at the 1 st , 3 rd , 5 th and 7 th day with CPAP on. Patients were monitored by a video camera to ensure they were sleeping in supine, alcohol and sedative drug were not permitted. Among them, 20 patients used auto-CPAP treatment at home were followed up, pressure and SpO2 profiles were down loaded at 2 and 6 month after treatment.

Results PSG testing revealed that auto-CPAP had the same effect as CPAP on AHI, SpO2 and sleep architectures. In the first week of treatment, the highest pressure level (hCPAP) required did not differ between days, and the every day level of hCPAP also did not differ from oCPAP. However, the mean CPAP pressure level (mCPAP) required decreased significantly (P < 0.01) except 5 th and 7 th , and the percentage of sleep time spent in low level of CPAP pressure increased, and in high level of pressure decreased across time. Compare to the pressure at 7 th day of treatment, long term follow-up did not find a significant change of hCPAP and mCPAP at both 2 and 6 months. All patients had good compliance to CPAP treatment, with mean of 5.3 ± 1.9 h/d and at least 6 ± 0.8 d/week usage. No significant BMI change was noticed. Conclusion We found a decreased mean CPAP pressure level during short term treatment, which could be attributed to REM sleep rebound. As both short term and long term hCPAP required by patient did not change, there appears to be not necessary to do re-titration in most of the OSAHS patients using CPAP. Methods 31 OSAHS patients after UPPP surgery were recruited and studied more than 12 months after the procedure. Among them 24 patients were treated with classical UPPP (cUPPP), which removes all of uvula and part of soft palate. 7 had modified UPPP (mUPPP), keeping part of uvula. The control group was 31 age, BMI and AHI matched, newly diagnosed OSAHS patients without prior treatment. A manual titration of CPAP was performed during both NREM and REM sleep in all 62 patients. Patients were studied in the supine posture. When significant mouth air leakage occurred and/or titration worsened with an increase of CPAP pressure, the pressure level was considered as the highest CPAP (hCPAP) patient can tolerate in that sleep stage. The CPAP machine used for titration could produce a highest pressure of 20 cm H 2 O. Results 74% (23/31) of UPPP patients had less than 50% decrease in AHI, and 84% (26/31) of the 31 patients still had AHI >15 (range: 16-110) during postoperation PSG test, and most of needed therapy. All of the untreated OSAHS patients could tolerate 17-20 cm H 2 O of CPAP during sleep. None had severe mouth air leak before an optimal pressure reached. In contrast, four of the surgery group failed to respond to CPAP treatment during both NREM and REM sleep, and one more during REM sleep. All of the seven patients who had a mUPPP could tolerate CPAP. One of the three tested both before and after surgery failed to CPAP treatment after surgery during REM sleep. Conclusion In many patients, UPPP may compromise the ability to apply nasal CPAP as a subsequent therapy. Methods There were 89 patients with OSAS who were confirmed by full nocturnal polysomnography. Height, weight, body mass index (BMI), neck circumference (NC), neck length (NL) of patients were measured. These patients were divided into mild group [5 < apnea + hyponea index (AHI) < 20] and moderate-severe group (AHI > 20). Correlation analysis between the AHI and weight, BMI, NC, NL, NC/NL was performed. Thirty two non-apneic healthy volunteer served as controls (AHI < 5), student's t test was used to examine the differences in all variables between the apneic and non-apneic groups, and differences in variables between the mild and moderate-severe group.

The weight, BMI, NC, NC/NL in apneic patients were significantly higher than that in the controls, weight, BMI, NC in moderate-severe patients were also significantly higher than that in the mild patients. Simple correlate analysis showed that the AHI significant positive correlated with weight, BMI, NC and NC/NL in all patients. Multiple stepwise linear regression showed AHI had significant positive correlated with weight and NC/NL, and had significant negative correlated with nadir oxygen saturation in sleep (multiple correlation coefficient r = 0.7795, P < 0.0001).

Conclusions This study showed that patients with OSAS are more obese and have thicker and shorter necks, it suggests that weight, neck circumference and neck length play an important role in the pathogenic factor of patients with OSAS.

MC JORGE II, AR ABE, GR REFRE, PV ZAMORA, SG SILVERIO

Background and Objectives OSA prevalence in the Western countries is estimated at 5% and around 2%-5% among studied Asian countries. In our country, the prevalence is unknown. This study was to determine the prevalence of OSA symptoms and EDS among Filipino office workers. Methods Consecutive office workers with 8 hour daytime daily work schedules who were consulting in their medical clinic were evaluated for OSA symptom and EDS from January-March 2004. The questionnaire asked about common signs and symptoms, risk factors, co-morbid conditions and demographic characteristics related to OSA. Standardized Epworth Sleepiness Scale (ESS) was used to detect daytime sleepiness. Results A total of 296 subjects were screened. None of the subjects had complaints referable to OSA nor EDS. Despite this, 38.1% had an Epworth score of at least 10. Snoring, bothersome snoring, BMI > 25 and presence co-morbid cardiovascular disease were also highly prevalent. 13.9% of the subjects had 5 or more OSA symptoms. Males had more OSA symptoms as compared to females. Likewise, the elderly age group had more OSA symptoms as compared with the younger age group. Thirty patients underwent overnight polysomnograph test, and 23/30 had evidence of OSA with six belonging to the severe category.

The study was consistent with the prevalence of OSA symptoms worldwide. OSA is an unrecognized problem in the primary care setting in the Philippines. Physicians should suspect and inquire about OSA and EDS among their patients. Further studies are recommended to clearly define its distribution and understand the actual burden of OSA in our country. (Supported by a research grant from JICA)

A RUS ANIDA, AC WILSON, SM STICK

Background Pulse Transit Time (PTT) had shown promise in the diagnosis of sleep disordered breathing as a measure of respiratory effort and arousal in adults and children. There is evidence in adults and children that PTT arousal is more sensitive than cortical arousal in measuring obstructive events. However the exact role of PTT arousal in the diagnosis of sleep disordered breathing in children is yet to be defined. Objective To explore the relationship between the autonomic arousal index as detected by PTT and cortical arousal index as detected by electroencephalograpy (EEG) with the respiratory events in children evaluated for obstructive sleep apnoea. Methods Tenty-two children aged between six and fourteen years old referred for further evaluations of possible obstructive sleep apnoea to the Sleep Unit at Princess Margaret Hospital between March to July 2004 were recruited. Standard overnight polysomnography was performed using Compumedic e-series and scored by standard criteria. PTT arousal was defined as reduction of more than 15 ms from baseline of greater than 5 seconds duration. Results Twenty-two patients consented for the study and fulfilled the inclusion criteria. Mean pulse transit time arousal index was significantly lower than the mean EEG arousal index. Most EEG arousals were not associated with PTT arousals. Similarly most PTT arousals occurred independently of EEG arousal. Respiratory events were more likely to result in EEG arousal than PTT arousal. Conclusion PTT arousal does not appear to be a more sensitive marker of arousal than EEG arousal. Furthermore PTT arousals do not relate to respiratory events. Further work is needed to determine the significant of PTT arousal in children. Background In 1993 Hoffstein and associates have reported the prediction formula for starting level of continuous positive airway pressure (CPAP) in patients with obstructive sleep apnea syndrome (OSAS). The ability to predict CPAP is likely to be of value in clinical sleep laboratory when allowing the optimal CPAP level to be achieved with only a few changes. However, the utility of the prediction formula has not been well assessed in Japanese patients with OSAS and some potential differences of the variability in predictive CPAP between Asian and Caucasian could be expected.

Methods Two-hundred-ninety patients who were diagnosed as OSAS and candidates of CPAP treatment in our clinical sleep laboratory were included in this study. Patients underwent the standard overnight polysomnography (PSG), Epworth Sleepiness Scale (ESS) test, pulmonary function measurements (lung volumes, airway resistance and flow-volume curves), and anthropometric measurements (including body mass index (BMI) and neck circumference (NC). First, we compared between the optimal CPAP levels determined from CPAP titration and the starting pressure levels were determined from the prediction formula utilizing BMI, NC and apnea/hypopnea index (AHI). Second, we examined which factors contribute to the variability of predictive CPAP level in our patients with OSAS.

Results Adequate CPAP titrations were achieved in 288 patients. There was significant difference between the levels of optimal CPAP and the levels of predictive pressure determined from the prediction formula which has been reported (7.4 ± 1.6 cm H 2 O v.s. 9.3 ± 3.0 cm H 2 O for the predicted and measured values, respectively). Multiple linear regression analysis was used to further examine the determinants of predictive CPAP level, which was the dependent variable. In our patients group maximum r 2 methods revealed that the best threevariable subset was the one that included BMI, Time-S p O 2 < 90%, and ESS. This subset of variables resulted in r 2 = 0.42. Conclusion We conclude that the prediction formula which has been reported in Caucasian patients with OSAS may not fully explain the measured value of CPAP levels in our patients group. It may be caused by the potential differences between the two ethnic groups (Asian v.s. Caucasian patients with OSAS). Background Obesity, in particular abdominal obesity, is an established determinant of insulin resistance. There is growing evidence supporting a causal contribution of OSA on insulin resistance (1,2), independent of obesity. This prospective cohort study further investigated the relationship of abdominal fat, insulin resistance and sleep apnea. Methods Subjects were recruited from our sleep laboratory. Demographic and anthropometric data were collected. Overnight 16-channel polysomnography was performed. Insulin resistance was estimated by the Short insulin tolerance test (SITT), and was indicated by the rate of decline in blood glucose (K constant). and polysomnographic findings were collected. MRI scan of abdomen was also performed to estimate the amount of abdominal fat.

The major dependent outcome variable was the apnea-hypopnea index (AHI). OSA was defined as AHI ≥ 5. Independent measures were body weight (BW), BMI, waist circumference, abdominal fat analysis (MRI imaging), and the lipid profile. Significant linear relationships of AHI with BW, BMI, abdominal fat, HDL-cholesterol and Apolipoprotein B were demonstrated (P < 0.05). Multiple linear regression analysis showed that obesity measured by either abdominal fat (P < 0.001) or waist circumference (P < 0.001) or BMI (P < 0.001) as well as Apolipoprotein B levels (P < 0.05) were independently associated with AHI. Conclusion In this cohort of Chinese patients, Apolipoprotein B was associated with AHI as controlled for obesity, and it may contribute to atherosclerosis in OSA. Objective Sleep disordered breathing is a well recognized complication in patients with neuromuscular disorders. A prospective study was performed to assess the prevalence of sleep disordered breathing and identify risk factors for development of this complication in a group of paediatric patients with neuromuscular disorders. Method We performed multidisciplinary assessment of 17 patients with various neuro-muscular disorders who have been followed up at the Department of Paediatrics of a regional hospital. The aspects assessed included sleep polysomnography, pulmonary function (spirometry, maximum inspiratory and expiratory pressure), venous blood gas, echocardiography for pulmonary hypertension, nutritional status (median weight for height), mobility and functional disability. 

GR REFRE, MC JORGE II, TC CAPALONGAN Department of Medicine, Section of Pulmonary Medicine, University of the Philippines-Philippine General Hospital, Manila, Philippines

Background and Objectives Prevalence of excessive daytime sleepiness (EDS) is common among several occupations including medical students and medical trainees. This is the first study in our country in determining the level of sleepiness and sleeping patterns among adults who work in a health related facility using the Epworth Sleepiness Scale (ESS) and a self administered questionnaire on sleep habits. Methods All residents in various departments were given a self-administered questionnaire. The questionnaire included the ESS and questions on sleep habits and patterns. A cohort of 45 age-matched office workers who worked during daytime (8 AM and 5 PM) served as control.

Results 277 residents and 45 controls responded. Prevalence of excessive daytime sleepiness (EDS), i.e. mean ESS score >10, among residents was 90.5% compared to control, 26.7% (P = 0.000). Mean ESS score was 12.3 compared to control with mean ESS of 8.3, (P = 0.000). Sleepiness was most noticeable among lower year level residents (P = 0.007) and those with prolonged duties (P = 0.000). No statistical difference was noted across departments, age and sex. 73.6% had sleep duration of less than or equal to 6 hours. 57% had 1-3 awakenings per night mostly secondary to emergency room/ward calls. 87.9% of the residents felt sleepy during the day and 87.2% still feel unrefreshed after a night's sleep. 45.7% took naps while 67.2% had at least 2 cups of methylxanthine containing beverages within the 24-hr period. 61% did not need any methods to help them fall asleep; however, 1.4% reported use of sleep aid medications. None took any medications to keep them awake.

Conclusions EDS among the residents was highly prevalent. EDS was primarily brought about by sleep loss secondary to prolonged working hours and sleep fragmentation leading to circadian rhythm disturbances. However, fatigue and other stresses could not be discounted. Further studies are recommended to test for cognitive function, psychomotor performance and other work related learning and personal health variables to further address the sleep deficiency experienced by our residents. These may have an impact on one's performance. Background and Objective Epidemiological data have demonstrated that obstructive sleep apnea-hypopnea syndrome (OSAHS) is associated with coronary heart disease (CHD). However, the link between OSAHS and CHD has not been clarified. It is necessary to investigate the associated of the changes of endothelial function in OSAHS with CHD and efficacy of nasal continuous positive airway pressure (nCPAP) therapy.

Methods 80 subjects recruited into 4 subgroups. 20 control subjects, 20 OSAHS, 20 CHD, and 20 OSAHS with CHD. Each of them underwent PSG, nitric oxide (NO), endothelin-1 (ET-1) detection at run in and 7 day nCPAP treatment (mainly used in OSAHS and OSAHS with CHD). Results (1) Comparison the control subjects with CHD group, there were significantly higher concentration of ET-1 and lower value of MSaO 2 , concentration of NO, ratio of NO/ET-1 in both OSAHS and OSAHS concomitant CHD group (P < 0.01). There were no significant difference of the sleep apnea indices between OSAHS and OSAHS with CHD (P > 0.05). However, in the group of OSAHS with CHD the plasma ET-1 was significantly higher, the serum NO and ratio of NO/ET-1 were significantly lower than those in the group of OSAHS (P < 0.01).

(2) The concentration of serum NO and the ratio of NO/ET-1 in the group of OSAHS were positively correlated with MSaO 2 (r = 0.519, r = 0.504, P < 0.05). The concentration of plasma ET-1 was inversely correlated with MSaO 2 (r = -0.457, P < 0.05). There were no significant correlations between the concentrations of plasma ET-1 or the serum NO with the indices of sleep apnea in the group of OSAHS concomitant CHD.

(3) In the groups of OSAHS and OSAHS concomitant CHD, MSaO 2 , NO and NO/ET-1 after nCPAP treatment were significantly higher than those before treatment, ET-1 was significantly lower than that after treatment (P < 0.01); In group of OSAHS the change of NO, ET-1 and NO/ET-1 were significantly positively correlated with the changes of MSaO 2 (r = 0.547, r = 0.558, r = 0.577, P < 0.05).

Conclusion (1) The endothelial function was more significantly impaired in patients with OSAHS and CHD. The impaired endothelial function may be one of the main mechanisms that developed and deteriorate CHD in OSAHS patients.

(2) The vascular endothelial dysfunction can be ameliorated by short-term effective nCPAP treatment. The change of endothelial function is correlated with those of the sleep apnea and the nocturnal hypoxemia. Treatment with nCPAP may be helpful for the prevention from CHD in OSAHS patients and for control of CHD.

009 043

Rationale To determine the use of desensitization in the treatment of patients with immunologically mediated reaction to antituberculosis drugs and to describe the reactions manifested by the patients during desensitization with antituberculosis drugs. Methods Patients diagnosed with tuberculosis referred to the allergy clinic for desensitization after developing adverse drug reaction to the antituberculosis drugs regimen. During the desensitization procedure patients were closely monitored for changes in vital signs and appearance of adverse reactions until the desired dose of the drug tested was achieved. Reactions observed were managed base on its severity. Results There were 15 episodes of drug desensitization on 5 cases, 2 females and 3 males. The age ranges from 32 to 78 years old. Four episodes of desensitization to isoniazid, 3 episodes for pyrazinamide, 4 episodes for ethambutol, 2 episodes for rifampicin, 1 for streptomycin and 1 for ofloxacin were done. The reactions noted during the desensitization were fever, skin itchiness, rash, joint pains, elevation in blood pressure and vomiting.

Conclusions Drug desensitization is one modality of treatment for patients with immunologically mediated reaction to antituberculosis drugs. Desensitization entails some risks and reactions are observed but if managed well will give a successful procedure ensuring an optimal treatment for antituberculosis regimen.

Background The drug-drug metabolic interaction of antituberculous drugs with other medication is becoming a very common problem in clinical practice, which can leads to poor prognosis and serious adverse reaction. Cytochrome P450 enzyme system is key enzymes dealing with drug metabolism, among which isoenzyme CYP1A2 and 3A4 take part in 80% drug's metabolism. As we know, isoniazid is a inhibitor of some Cytochrome P450 isoenzymes, but rifampicin is a inductor of some Cytochrome P450 isoenzymes, yet what is the combination of the two drugs? Furthermore, most of these informations were from animal experiments. As there are species differences between animal and human, are the changes in human beings same as in animal? It is difficult to study the influence of the combination of isoniazid and rifampicin to the activity of Cytochrome P450 of human being, because of the reasons of medical ethics. Recent studies showed, human primary hepatocytes are the most similar system to liver in vivo in studying drug metabolism and hepatotoxicity. So we took the primary human hepatocytes as trial system to study the influence of isoniazid and rifampicin to the activities of CYP1A2 and 3A4, and the influence of these two drugs to the expression of mRNA of these two CYP450 isoenzymes. Objective To study the influence of either single or combinative using isoniazid and rifampicin to the activity of CYP1A2 and 3A4. (2) To observe the influence of either single or combinative using isoniazid and rifampicin to the expression of CYP1A2mRNA and 3A4mRNA, to explore the mechanism of influence of these two drugs to the activity of CYP1A2 and 3A4. Methods Part one: Hepatocytes were isolated from human liver, cultivated for 3 days. Then added isoniazid (25 mm; 50 mm), rifampicin (12.5 mmL; 25 mm), and both of them (for CYP1A2: rifampicin 12.5 mm & isoniazid 50 um, rifampicin 25 um & isoniazid 50 mm; for CYP3A4: rifampicin 12.5 mm & isoniazid 25 mm, rifampicin 25 um & isoniazid 25 um, rifampicin 25 um & isoniazid 50 mm), respectively. After cultured for 2 days added substrate (for CYP1A2: phenacetin; for CYP3A4: testosteone), 1 h later measured metabolite of them (for CYP1A2: acetaminophen; for CYP3A4: 6b-testosteone) with HPLC to assess the activity of CYP1A2 and 3A4. Part two: Hepatocytes were isolated from human liver, cultivated for 3 days. Then added isoniazid, rifampicin and both of them, respectively, at the concentration of which as those in Part one. With RT-PCR technique detected the expression of CYP1A2mRNA and 3A4mRNA, to explore the mechanism of these drug's influence to activities of CYP1A2 and 3A4. Results Part one: The activity of CYP1A2 in isoniazid groups with the concentration 25 mm and 50 mm was lower than that in control group (P < 0.01), but not less than 50% of that in control group. The activity of CYP1A2 in rifampicin group with the concentration 12.5 mm was higher than that in control group (P < 0.05), but was not 2 folds or more of that in control group. There was no statistical difference in activities between rifampicin with 25 mm and control groups (P > 0.05); The activity of CYP1A2 of combinative the groups with isoniazid and rifampicin was lower than that of the control group (P < 0.01), but was not less than 50% of that of control group. The activity of CYP3A4 in isoniazid groups with the concentration 25 mm and 50 mm was less than 50% of that in control group. The activity of CYP3A4 in rifampicin groups with the concentration of 12.5 mm and 25 mm was 13.18 and 9.59 times of that in control group, respectively. The activity of CYP3A4 in the combinative groups of isoniazid and rifampicin with 3 different concentration was 9.44, 6.19 and 5.48 times of that in the control group, respectively, but they were lower than those in rifampicin group with corresponding concentration (P < 0.05). Part two: Isoniazid with the concentration of 25 mm and 50 mm groups showed lower expression of CYP1A2mRNA than that of control group (P < 0.05), but there was no difference in expression of CYP1A2mRNA between isoniazid with the concentration of groups 25 mm and 50 mm (P > 0.05). There was also no difference in expression of CYP1A2 between rifampicin groups with the concentration 12.5 mm or 25 mm and control group. The expression of CYP1A2mRNA in the combinative groups of isoniazid and rifampicin with the concentration of R12.5 mm & I50 mm, R25 mm & I50 mm group was lower than that in the control group (P < 0.05). There was no difference of the expression of CYP1A2mRNA between the combinative groups of isoniazid and rifampicin and single isoniazid groups with corresponding concentration (P < 0.05). The expression of CYP3A4mRNA in the isoniazid groups with the concentration of 25 mm and 50 mm that was lower than that in control group (P < 0.05). The expression of isoniazid with 50 mm group was lower than that in 25 mm group. The expression of CYP3A4mRNA in the rifampicin with the concentration of 12.5 mm and 25 mm group was higher than that in control group (P < 0.05). There was no difference of the expression level between rifampicin groups with 12.5 mm and 25 mm. The expression of CYP3A4mRNA in the three groups with different combinative concentration was higher than that in control groups (P < 0.01). But the expression in the combinative groups was lower than that in single rifampicin group with corresponding concentration (P < 0.05, P < 0.01 and P < 0.05, respectively). Conclusion (1) When isoniazid and rifampicin are used alone or combinatively in the range of maximum clinical blood concentration, they have not significant might inducting and inhibiting action to CYP1A2. (2) The combination of isoniazid and rifampicin cold induce the activity of CYP3A4, but the inducing level was lower than that induced by single rifampicin with the same concentration. (3) The drugs might influence the activity of CYP3A4 by the way of regulation the expression of CYP3A4mRNA.

Objective To investigate the protective effect of CpG oligodeoxynucleotides (ODN) on mice infected with mycobacterium tuberculosis. Methods 96 female BALB/c mice were randomized into 4 groups, namely CpG ODN treatment (A group), control ODN treatment (B group), infectious control (C group) and healthy control (D group) (n = 24). A group and B group were treated once with CpG ODN or control ODN (30 mg/mouse) intraperitoneally 2 weeks before infection with mycobacterium tuberculosis. 1 ¥ 10 6 bacili in a volume of 300 ml saline were injected into the tail vein of mice from A group, B group and C group. The 12 mice from each group were sacrificed at 3 weeks postinfection. The expression of Toll-like receptor 9mRNA and ctokines was detected by RT-PCR. The rest 12 mice from each group were monitored for 60 days to account the motality. Results CpG ODN could lengthen survival time of mice infected with mycobacterium tuberculosis, and decrease inflammation in lung tissue, and reduce mycobacterial outgrowth in lungs and spleens. The expression of TLR9mRNA in lungs and spleens of mice infected with mycobacterium tuberculosis was higher than that in healthy ones and CpG ODN didn't affect the expression of TLR9mRNA. Furthermore CpG ODN could increase the expression of IFN-g mRNA and IL-6 mRNA and depress the expression of IL-4 mRNA and IL-10 mRNA in lungs and spleens tissue. Conclusions CpG ODN could increase the immunity of mice infection with tuberculosis through up-regulate Th1 immunity and downregulate Th2 immunity. Backgroud Nontuberculous mycobacterial pulmonary disease may occur in patients with no underlying lung disease and no known immunodeficiency. The purpose of this study was to evaluate a potential role of natural-resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms for human susceptibility to the nodular bronchiectatic form of nontuberculous mycobacterial pulmonary disease. Methods A case-control study was performed to compare the frequency of several polymorphisms in the NRAMP1 gene in 41 adult patients with nontuberculous mycobacterial disease (18 patients with M. avium complex infection and 23 patients with M. abscessus infection) and 53 healthy controls. Results Heterozygotes at INT4, D543N, and 3¢UTR were observed at significant higher frequencies in patients with nontuberculous mycobacterial disease than in controls (P = 0.033, P = 0.014, and P = 0.002, respectively). The ORs were 2.61 (95% CI 1.07-6.37) for INT4 G/C, 4.31 (95% CI 1.26-14.78) for D543 G/A, and 7.15 (95% CI 2.12-24.18) for 3¢UTR TGTG+/del. Subjects who were heterozygous for two NRAMP1 polymorphisms in INT4 and D543N were particularly overrepresented among those with nontuberculous mycobacterial disease, as compared with those with the most common NRAMP1 genotype (odds ratio 10.88, 95% CI 1.18-100.45, P = 0.035). There were no significant differences between the patients with M. avium complex infection and those with M. abscessus infection. Conclusion These findings suggest that the NRAMP1 genetic polymorphisms are associated with human susceptibility to nontuberculous mycobacterial pulmonary disease. Objective To investigate the in-vitro interferon-gamma (IFN-g) release assay based on different Mycobacterium tuberculosis (M. tb) antigens in the diagnosis of tuberculosis and of latent tubercular infections, and to compare it with the tuberculin skin test (TST). Methods All subjects were divided into three groups: group 1, patients with tuberculosis (n = 62); group 2, healthy (n = 27); group 3, patients with cancer (n = 29). The PBMC were cocultured for 5 days with different antigens: purified protein derivitives (PPD) of tuberculin, early secretary antigenic target 6 kDa protein (ESAT-6), 38 kDa antigen. The protein levels of IFN-g were detected by ELISA. Total RNA was extracted from the PBMC after 5 days culture. The expression of the IFN-g mRNA was detected by RT-PCR. Results For healthy control, the TST is positive related to the history of BCG vaccination and the contact degree with sputum-positive TB patients (P = 0.047, P = 0.041, respectively). But the ESAT-6 based IFN-g release assay is only significantly related to the contact degree with sputum-positive TB patients, and there is no significant positive relation with the history of BCG vaccination. The invitro IFN-g release level is highest in TB patients and is lowest in cancer patients. The sensitivity and the specificity of the IFN-g release assay based on 38 kDa are 64.9% and 89.3% for the diagnosis of tuberculosis. The expression of IFN-g mRNA was detected only in part of subjects using RT-PCR, and it did not correlated with the production of the IFN-g protein.

The IFN-g release assay based on ESAT-6 may be better than TST when diagnosis of the latency of M. tb. The IFN-g release assay based on 38 kDa may be a promising reagent when diagnosis to the tuberculosis. Expression of the mRNA for IFN-g does not parallel the production of the IFN-g protein. Objective To evaluate the effects of CpG oligonucleotide on the production of interferon-g (IFN-g), interleukin (IL)-12 and IL-10 in peripheral blood mononuclear cells (PBMC) in patients with active pulmonary tuberculosis (PTB).

Methods PBMC was isolated from 20 patients and 20 healthy donors, and then cultured in RPMI-1640 with non stimulator (control), PPD (10 ng/mL), CpG ODN (2 nmol/mL) and CpG ODN plus PPD, respectively. The expression of IFN-g mRNA, IL-10mRNA and IL-12mRNA in PBMC were detected by real time-PCR. Results In control group, the expression of IFN-g mRNA and IL-12mRNA in patients was lower than that in healthy donors (P < 0.01, P < 0.05), the expression of IL-10mRNA in patients was similar to that in healthy donors (P > 0.05). In PPD group, the expression of IFN-g mRNA, IL-12 mRNA and IL-10 mRNA in patients were higher than that in control groups (P < 0.01, P < 0.01, P < 0.01), but the expression of IFN-g mRNA in patients was lower than that in healthy donors (P < 0.01), there was no defference in the expression of IL-12mRNA between in patients and in healthy donors (P > 0.05), the expression of IL-10mRNA in patients was higher than that in healthy donors (P < 0.01). In CpG ODN group, the expression of IFN-g mRNA and IL-12 mRNA in patients were higher than that control groups (P < 0.01, P < 0.01), the expression of IFN-g mRNA and IL-12mRNA in patients was lower than that in healthy donors (P < 0.01, P < 0.05), the expression of IL-10mRNA in patients was similar to that in control groups and in healthy donors (P > 0.05, P > 0.05). In CpG ODN plus PPD group, the expression of IFN-g mRNA and IL-12 mRNA in patients were higher than that control groups (P < 0.01, P < 0.01), the expression of IFN-g mRNA was lower than that in healthy donors (P < 0.01), the expression of IL-10mRNA in patients was similar to that in control groups and in healthy donors (P > 0.05, P > 0.05). Compared with PPD group, in CpG ODN plus PPD group, the expression of IL-12mRNA in patients was increased (P < 0.05), while the expression of IL-10mRNA in patients was impressed (P < 0.01), and similar to that in healthy donors (P > 0.05).

Conclusions (1) There are decreased ability of production of Th1-type cytokines (IFN-g and IL-12) and increased potential for Th2-type cytokine (IL-10) in PBMC from patients with PTB.

(2) CpG ODN could increase the production of IFN-g and IL-12, and decrease the production of IL-10. 

Background Incidence of tuberculosis is significantly higher in the elderly, and 2-3 times greater in diabetics. Singapore has one of the fastest aging population in the region, and prevalence of diabetes is steadily rising. Aim To analyse the clinical profile and estimate the prevalence of diabetes mellitus in smear positive pulmonary tuberculosis. Methods A prospective study was carried out on 58 adult patients, admitted to a general medical department with chronic cough, new pulmonary infiltrates and positive sputum smear for acid fast bacilli. There were 49 males and 9 females. 53% (31patients) had diabetes mellitus, the national prevalence is 4.7% among residents aged 18 years and above. 40% of Chinese and 60% of Malays and Indians suffered from diabetes. Among the 31 with diabetes, 61% were Malays, 29% Chinese and 10% Indians. One third of the patients were aged 60 years and above and 68% were diabetic, vs. a national prevalence of 20.7% in those aged 65 and above. Chest X-ray revealed, far advance disease in 48% of diabetics and 77% of non diabetics, and cavities were seen in 74% and 70% of diabetics and non diabetics, respectively. Conclusions The high prevalence of diabetes mellitus, especially among the elderly is a cause for grave concern. Public education and greater awareness among the health care providers is necessary, for early diagnosis and successful containment tuberculosis in Singapore. Objective To investigate the characteristic and its clinical value of tumor necrosis factor-a and its receptor, interleukin-1b and its receptor in serum and bronchoalveolar lavage fluid (BALF) in patients with pulmonary tuberculosis and to determine the role of them in the immunopathogenesis of tuberculosis. Methods The concentrations of TNF-a, sTNF-R I, IL-1b and IL-1R were measured using sandwish ABC-ELISA method in serum and BALF of 46 cases of active tuberculosis and 21 cases of inactive tuberculosis and in the serum of 20 cases of normal control. Meanwhile the above-mentioned cytokine levels in serum and BALF of 19 cases of active tuberculosis were followed up.

The concentrations of TNF-a, sTNF-R I, IL-1b and IL-1R and TNF-a/sTNF-R I ratios in the serum were significantly higher in active tuberculosis and inactive tuberculosis group than in normal control group (P < 0.01~0.05). The TNF-a, sTNF-R I IL-1b and IL-1R levels and TNFa/sTNF-R I ratios in the serum and BALF of active tuberculosis group were significantly higher than those of inactive tuberculosis group (P < 0.01~0.05). The concentrations of TNF-a, sTNF-R I and IL-1b and TNF-a/s TNF-R I ratios in the serum were significantly lower in cavernous tuberculosis group than in non-cavernous tuberculosis group (P < 0.01~0.05), while the TNF-a, sTNF-R I, IL-1b and IL-1R levels and TNF-a/sTNF-R I ratios in BALF of cavernous tuberculosis group were significantly higher than those of non-cavernous tuberculosis group (P < 0.01~0.05). After 2 months' antituberculosis treatments, among 19 cases, the TNF-a, sTNF-R I, IL-1b and IL-1R levels and TNF-a/sTNF-R I ratios in the serum and BALF of 16 cases were significantly lower than those of the beginning of treatments (P < 0.01~0.05), while there were no significant changes of the above-mentioned cytokine levels in the serum and BALF of the other 3 cases.

Conclusion TNF-a, sTNF-R I, IL-1b and IL-1R were likely to be involved in the immunopathogenesis of tuberculosis. Detection of TNF-a, sTNF-R I, IL-1b and IL-1R levels in the serum and BALF was helpful to understand the activity of disease, determine the clinical pattern of disease, assess the prognosis of disease and monitor the therapeutic effect in patients with pulmonary tuberculosis. Background The resurgence of tuberculosis, especially the development of multidrug resistant tuberculosis and dubious efficacy of BCG vaccine embarrassed the treatment and prevention-two keystones of tuberculosis control. The completion of whole genome of H37Rv is a rich mine. We can explore the resource by updated new technology to find better molecular targets for rapid diagnosis and treatment of tuberculosis. Methods The newly developed molecular approach called in vivo induced antigen technology (IVIAT) has suggested to be very useful to identify microbial genes expressed specifically during human infections. M. tuberculosis would turn on the expressions of battery of genes that would be advantageous against human immune system. We use the IVIAT to screen the recombinant M. tuberculosis expression library constructed in pET30 vector. The tuberculosis patients sera as probe to screen the library. Results By using tuberculosis patients sera to screen M. tuberculosis expression library, we got several positive reaction clones. The sequence of these clones were determined and BLASTed for homologue. One positive clone dedicated to one ORF rv3878. The rv3878 appear to be interesting as the ORF is present in a 9.5 kb DNA fragment containing esat6 locus. This fragment denoted as RD1 region is present in M.tb complex but absent from M.bovis BCG and most environmental mycobacteria. Objective To learn the relation of interleukin-15 (IL-15) and several cytokines (IFN-g, TNF-a, IL-4 and IL-10) secreted by peripheral blood mononuclear (PBMC) cells from tuberculosis patients. Methods PBMCs were isolatet and devided into four groups: RPMI1640 group, PPD group, PPD + rhIL-15 group, PPD + Anti-IL-15 group. The levels of IFN-g, TNF-a, IL-4 and IL-10 were detected with ELISA in supernatants.

The levels of IFN-g and TNF-a were lower in patients than those in healthy groups, the contents of IL-4 and IL-10 were not different betweent the two groups. Compared with PPD, rhIL-15 effectively enhanced the production of IFN-g and TNF-a, but decreased the production of IL-4 and IL-10 in the patients and healthy individuals. Anti-IL-15 inverted the effect of IL-15. The levels of IFN-g and TNF-a in type II pulmonary tuberculosis were lower than those in type III, but IL-4 and IL-10 were higher than those in type II. Conclusion IL-15 could enhance Th1 cytokine (IFN-g) and proin flammation (TNF-a), and inhibite IL-4 and IL-10, IL-15 could regulate the balance of Th1 and Th2 cytokines. IL-15 could be an important cytokine for tuberculosis immunetherapy. Background Rapid detection of resistant to rifampin is important, the conventional method of determining susceptibility to antituberculosis drugs by plating on solid medium requires a long time. The purpose of the present study was to evaluate the significance of quantitative analysis of Mycobacterium tuberculosis mRNA in the test of susceptibility of Mycobacterium tuberculosis strains to rifampin. Methods The susceptibility to rifampin of 53 clinical isolated strains were test by the percentage of Mycobacterium tuberculosis 85B mRNA copies before and after the use of rifampin, and 1% and 10% were used as the standards for drug resistance, which was compared with absolute concentration method. Among them, 29 were rifampin resistant strains and 24 rifampin sensitive strains.

Results When the concentration of rifampin was 1 mg/mL, and 1% and 10% were used as standards, the agreement of quantitative analysis of Mycobacterium tuberculosis mRNA with the absolute concentration method was 70% and 81%, respectively, and the sensitivity of quantitative analysis of Mycobacterium tuberculosis mRNA was 100% and 100%, respectively, while the specialty of quantitative analysis of Mycobacterium tuberculosis mRNA was 33% and 58%. When the concentration of rifampin was 2 mg/mL, the agreement of quantitative analysis of mRNA with the absolute concentration method was 85% and 93%, respectively, and the sensitivity was 100% and 97%, respectively, while the specialty was 67% and 88%, respectively. When the concentration of rifampin was 4 mg/mL, 1% the agreement was 93% and 93%, and the sensitivity was 93% and 93%, while the specialty was 92% and 92%. Conclusion Quantitative analysis of Mycobacterium tuberculosis mRNA was a rapid, sensitive method in rifampin resistance screening. We suggest that the critical concentration of rifampin be 2 mg/mL, and the critical proportion of mRNA copy be 10%. Background CDC suggests that the result of sputum culture at 2 months after starting the initial treatment for pulmonary tuberculosis is an important factor to decide the continuation treatment period. But it is unknown which of two culture systems, solid or liquid medium, should be used. This study's aim is to compare the characteristics of liquid medium system with those of solid medium system after starting the initial treatment, using MB/BacT system and the Japanese conventional egg-based Ogawa medium. Methods Retrospective study of 50 patients (median age 59.2 ± 20.1 year, 37 males), who received chemotherapy for pulmonary tuberculosis, and whose sputum culture results were monitored with both MB/BacT system and Ogawa medium for more than 2 months after starting treatment. Based on the culture results, we examined age, treatment-inhibiting factors to tuberculosis, treatment regimens, and radiographic findings among the groups.

Results At 2 months after starting the initial treatment, 6 patients (12%) (group 1) were positive with both MB/BacT system and Ogawa medium, and 7 patients (14%) (group 2) were positive with only MB/BacT system, and 37 patients (74%) (group 3) were negative with both. There was no significant difference in age, treatment regimens, and existence of cavitation among three groups. But group 1 and 2 had more treatment-inhibiting factors significantly than group 3 and group 1 and 2 had a tendency to have more spreading shadow in chest radiograph than group 3. Conclusion In this study 7 patients (14% of total), who were positive with only MB/BacT system at 2 months after starting the initial treatment, had some similar characteristics to patients who were positive with both. With only conventional solid medium, there is a possibility that we miss additional 3 month treatment after continuation phase. Objective To study the effect of Mtb on IFN-g signaling pathway in macrophages.

Method Humans and mice macrophages were isolated and infected by Mtb, condition medium were collected after a few days culture; Macrophage surface expression of FcgRI (human) or MHC II (mouse) was measured by flow cytometry to explore the effect of condition medium on macrophage response to IFN-g; Anti IL-6 antibody was added to the condition medium to further study the role of IL-6 in the infection of Mtb. Results Condition medium induced by Mtb infected macrophages can inhibit macrophage surface expression of FcRI or MHC II response to IFN-g. Surface expression of FcgRI in human macrophages cultured in condition medium were significantly decreased compared to those without condition medium; Significant differences were shown from 10% of condition medium to up (P < 0.05). Similarly, surface expression of MHC II in mouse macrophages cultured in the condition medium were significantly decreased compared to those without condition medium; Significant differences were shown from 2.5% condition medium to up. Condition medium cultured with anti IL-6 antibody can improve macrophage response to IFN-g; Surface expression of MHC II in mouse macrophages cultured in condition medium with anti IL-6 antibody were significantly increased compared to those cultured in condition medium alone (P < 0.05).

Conclusion Mycobacterium tuberculosis induced condition medium can inhibit macrophage surface expression of FcgRI or MHC initiated by IFN-g. At meanwhile, our results also show that Il-6 may play an important role in condition medium which can inhibits macrophage response to IFN-g, and indicate that IL-6 may have a novel function in the Mtb infection. Background In recent years, under the circumstances that epidemic situation is so serious and are rising multidrug-resistance cases, it will inevitably to hew out a new road to conquer Tuberculosis by the way of researching new medicine especially Chinese medicine. Methods Divide the 429 of patients, in the proportion of 3-1 with computer automatically, into two groups, the trial group and control group. The regimen of the newly diagnosed cases is 2HL 2 Z/4 HL 2 , the re-treated cases will be 3HL 2 ZV/5 HL 2 V. But all the patients of the two groups will take the medicine Qijiabuyinlifei capsule and placebo, respectively. Results 1 The sputum smear negative conversion rates of newly diagnosed cases at the end of initial phase were 81.0% and 61.9% in the trial group and control group (P < 0.05); and in the re-treated cases, the rates were 64.04% and 40.0% (P < 0.05).

2 At the completion of the therapy, the X-ray resolution rates were 79.0% and 48.7%, 58.3% and 30.3% in newlydiagnosed cases and re-treated cases, respectively, for the trial and control groups, the difference being significant (P < 0.01). 3 There were significant differences in CD4/CD8 in the trial and control groups in the newly diagnosed cases and re-treated cases (P < 0.01, P < 0.05). 4 The disappear rates of some tuberculosis symptoms in initial groups better than control groups both the newly diagnosed cases and re-treated cases.

The test result shows that the Chinese medicine Qijiabuyinlifei capsule takes a good effect: improve the sputum smear negative conversion rates, promote the focus to resolute, improve the immune function and lyses the symptoms. Besides this, safe and endurable as it is, it will be helpful to make the curative effect better. Background To investigate the effect of interleukin-7 on the production of interferon-g (IFN-g), tumor necrosis factor-a (TNF-a), interferon-4 (IL-4) and interferon-10 (IL-10) by peripheral blood mononuclear cells (PBMC) from patients with tuberculosis. Methods PBMCs were isolated and cultured from 20 patients with pulmonary tuberculosis and 15 healthy tuberculin reactors, respectively. Every sample was devided by RPMI1640, PPD, PPD + IL-7, PPD + anti-IL-7. After 72 h, the supernatants of culture PBMC were collected. The levels of IFN-g, TNF-a, IL-4 and IL-10 were measured by Enzyme-Linked Immunosobent Assay (ELISA).

Results Compared with the PPD group, IL-7 increased the production of IFN-g and TNF-a, but decreased the production of IL-4 and IL-10 in the patients and the healthy individuals; anti-IL-7 blocked this effect. The levels of IFN-g, TNF-a were lower in tuberculosis patients than those in the healthy controls (P < 0.05). The levels of IL-4 and IL-10 were not different between the two groups (P > 0.05). The levels of IFN-g and TNF-a were lower in disseminated tuberculosis patients than those in exudative tuberculosis cases (P < 0.05). The levels of IL-4 and IL-10 were higher in disseminated tuberculosis patients than those in exudative tuberculosis cases (P < 0.05).

Conclusion IL-7 could regulate the balance between Th1 and Th2 cytokines by inducing IFN-g, TNF-a production and inhibiting IL-4, IL-10 expression, which suggested IL-7 enhance the patient defense against the infection of tuberculosis. IL-7 could be useful for the treatment tuberculosis. Objectives A genetic tendency to Pulmonary thromboembolism (PTE) is referred to as inherited thrombophilia and it is undoubtedly a polygenic abnormality. The basis of our understanding of the thrombophilic state is that it arises from an imbalance between the procoagulant, anticoagulant and fibrinolytic components of the coagulation system, resulting in an increased tendency to thrombosis. In this casecontrol study, we aimed to determine the prevalence of the fibrinolytic parthway related gene polymophisms such as the insertion deletion polymorphsim of human tissue type plasminogen activator (TPA) gene, dimorphism in the plasminogen activator inhibitor (PAI-1) promoter 4G/5G polymorphsims in Chinese Han population and investigate whether they are associated with PTE. Methods The subjects consisted of 101 patients with PTE and 101 age and sexmatched healthy controls from the same geographic area. All patients were diagnosed by high probability of lung ventilation/perfusion scan and/or multi-slice CT pulmonary angiography as well as medical history and clinical manifestations. Genome DNA was extracted from whole blood using KI-phenol-chloroform. (1) Standard PCR were used to determine the prevalence of the insertion deletion polymorphsim of human TPA gene.

(2) Standard PCR, Denaturing high performance liquid chromatography (DHPLC) as well as sequences analysis were used for screening different single neucltide polymorphisms and genotype distribution of -675 4G/5G located in the promoter region of the PAI-1 gene. Results TPA gene: Frequencies of allele I and D in the controls were 0.515 and 0.485 respectively, the distribution of genotypes met the Hardy-Weinberg equilibrium. No significant differences were found in the frequencies of genotype II, ID and DD between patients and controls., nor in allele I and D. The incidences of DD genotype of TPA genes were slightly higher in cases than in controls. Background The pathogenic mycobacteria cause the deaths of millions of people every year. One of the reasons these pathogens are so successful is that they are able to invade and replicate within host macrophages, so study the difference in gene expression between the tuberculous granulation tissue and normal tissue nearby the lesions is important for selecting the candidate of TB bio-therapy. Methods We made cDNA microarray in which 626 immunogenesis genes were involved. The tuberculous lesions were in 11 osteoarticular tuberculosis, Paired mRNAs from normal osteoarticular tissue specimens from the same cases were labeled with different fluorochromes during cDNA probe synthesis in a reversetranscription reaction. The signal intensity of each spot was measured by laser scanner and gene expression was quantified as the tubercle-to-normal fluorescence ratio (T : N ratio). The gene was overexpressed when the T : N ratio was greater than 2.0 and underexpressed when the ratio was less than 0.5. Results Among 626 immunogenesis associated genes there were 66 genes difference significant, of which 52 expressed higher and 14 lower in tuberculous granulation tissue than that of the controls. Conclusion The numerous alterations of gene expression were present in tuberculous lesions of osterarticular tuberculosis specimens. These results increase the understanding of the mechanisms used by pathogenic mycobacteria to cause disease, the host response to these organisms and provide new insights for antimycobacterial intervention strategies. Poland's syndrome (PS) is a non-genetic congenital abnormality characterized by various degrees of unilateral anterior chest wall hypoplasia and a variety of associated anomalies. PS occurs in 1 : 30,000 live births. We observed 27 patients, 20 of whom we operated upon. The severity of cardio-respiratory impairment correlates with the severity of the syndrome. Reduction of thoracic volume due to depression deformity is observed in up to 25% of PS patients. Paradoxical chest wall movements are possible, especially during stress or pulmonary infection with a reduction of vital capacity to half, particularly in patients with rib defects, which occur in 20% of PS patients. Lung herniation occurs in 8% of PS cases. Radiography confirms unilateral hyperlucent lung and shift of the heart towards the unaffected side. Isolated dextrocardia is observed in 6%, and always in patients with left-sided PS. Bullous pulmonary disease and spontaneous pneumothorax were also described. Extreme forms of PS may be associated with diaphragmatic hernia. Surgical correction is usually carried out in children in two stages, and in adults in one stage. During surgery, unilateral ventilation of the opposite lung is recommended, as well as epidural anesthesia, which should be continued postoperatively. The rib defect and lung herniation are corrected by bony allo-or autografts or split rib grafts taken from the unaffected side. It may also be repaired with a prosthetic mesh patch sutured to the aplastic rib stumps in combination with the latissimus dorsi musculocutaneous flap. In female adult patients, prosthetic breast implantation is advisable. Background To identify the causes and clinical characteristics, and efficacy of various diagnostic approaches, we studied prospectively of the pediatric patients with chronic recurrent chest pain. Methods Prospective study of 122 patients with chronic recurrent chest pain from June 1998 to June 2003 were performed. Male and female ratio was 81 : 41, age 9.3 ± 3.1 year. A single chart including pain description (histoy) and pain questionaire, associated sympoms was used for systematic history taking. The patients were devided in two groups, Group A, before June 2001 (n = 70), Group B, after June 2001 (n = 52). Chest X-ray and ECG were checked in all patients. But, allergy test and echocardiography, 24 hour ECG monitoring, exercise test, pulmonary function test, gastrointestinal fiberendoscopy were performed selectively. Results Idiopathic origin was most common (32%). The remaining causes were psychogenic (23%) and exercise-induced asthma (20.5%), hyperventilation syndrome (9.1%), tachyarrhythmia (4.9%), cardiac (4.1%), pulmonary (3.3%), reflux esophagitis (2.5%) in order. The positive rate of allergy test (24%) was higher than that of cardiac examinations (11-20%). Exercise-induced asthma was more common than psychogenic causes after June, 2001. Various abdominal symptoms were accompanied in idiopathic and psychogenic chest pain, hyperventilation syndrome. Headache was more commonly found in psychogenic pain, but chest tightness and dyspnea were common in hyperventilation syndrome. About half of exercise-induced asthma had symptoms of allergic rhinitis. Conclusion In majority (72%) of exercise-induced asthma, the chest pain was induced or aggravated by exercise, and relieved by rest. Causative antigens were detected in 69.2% among group B patients with exerciseinduced asthma. Exercise-induced chest pain might be a first manifestation in allergy patients. So, we recommand allergy test for patients with recurrent chest pain that was inducing or aggravating by exercise. Hence, cardiac examinations such as echocardiography or 24 hour ECG monitoring could performed selectively case by case. Etiology of Chronic thromboembolic pulmonary hypertension (CTEPH) is largely unknown and may be heterogeneous, because there are several ethnic differences in clinical characteristics of CTEPH. Female predominance and a higher ratio of chronic to acute pulmonary thromboembolism have been reported in Japanese as compared to the USA. Because such ethnic differences may be controlled by genetic factors, we investigated HLA polymorphisms in Japanese patients with CTEPH. HLA typing by serological and/or DNA typing methods was performed (for HLA-A, B, DPB1, DRB1) in 80 patients and 678 controls, and the association of clinical characteristics with HLA alleles was studied. The frequencies of HLA-B*5201 (40% vs. 24%, odds ratio = 2.14, Pc = 0.037) and DPB1*0202 (19% vs. 6%, odds ratio = 3.41, Pc = 0.0014) were significantly higher in the patients. HLA-B*5201 positive patients showed significant female predominance. Total pulmonary vascular resistance and mixed venous oxygen tension were better in the HLA-B*5201 positives. In contrast, cardiac index and gas exchange parameters were worse in the HLA-DPB1*0202 positives. In the patients carrying HLA-B*5201 and/or -DPB1*0202, the frequency of deep vein thrombosis was significantly lower than the other patients. These observations suggested that both the susceptibility and clinical characteristics of CTEPH were controlled in part by the HLA-B and -DPB1 loci. Background Studying the incidence of cause of lung mass and differential diagnosis in adult cases was made, and value is finely known and exposed to all, but unfortunately such studies has not yet made on children especially in Iran. Materials and Method Studying a six years survey (1376-1382) on operated cases files in Dr. Sheikh Hospital, 37 cases under went surgery for lung mass. Results We found that hydatid cyst were 19 cases (51%); local empyema 6 cases (16%); lung abscess 4 cases (11%); congenital cystic adenomatoid malformation (CCAM) 4 cases (11%); Pleuropulmonary blastoma 2 cases (5%); pulmonary sequestration 1 case (3%) and askin tumor 1 case (3%). Conclusion We realize that the chief cause for lung masses in children in this study are infection (hydatid cyst Empyema and abscess) and congenital and neoplastic diseases are rare in lung masses in children.

Considering the statistical information in Iran and developed countries, we witness the importance of epidemiological factor of this disease in our country. Conclusion This is the first study from Pakistan on CT proven APE that shows the clinical and radiological characteristics and disease spectrum leading to APF and its out comes. High clinical suspicion and imaging techniques are required for early diagnosis and prompt treatment.

Objective To investigate the mechanism of lung injury associated with pulmonary thromboembolism (PTE) in rats.

Methods Experimental PTE models were induced by injecting emboli into external jugular veins of 28 rats. Other 28 animals received only saline served as shams. Blood serum and BALF were analysed for sVCAM-1, MMP-9 and VEGF at 2 h, 6 h, 24 h and 72 h after the models were induced. The total protein and cells count in BALF were examined. The expressions of MMP-9 and VEGF were evaluated by immunohistochemical staining.

Results (1) The embolic areas revealed inflammatory cells infiltration, widening and rupture of alveolar septation, dilation of capillary, alveolar collapse and pulmonary atelectasis under the microscope.

(2) The serum levels of MMP-9, sVCAM-1 and VEGF were elevated significantly in PTE group compared with that in sham.

(3) Compared with those of shams, the levels of TNF-a VEGF and MMP-9 in BALF elevated significantly. (4) The expression of the MMP-9 and VEGF expressed more positively in PTE group than those of sham group in lung by immunohistochemical stain. Conclusion Acute PTE could caused the release of VCAM, MMP-9 and VEGF in lungs, thus, leads to lung injury. All three cytokines contribute to the extravasation of inflammatory cells and protein and the processes of lung injury associated with PTE. Background Previous studies have shown that hypoxia induces IL-1b and TNF-a in a variety of cells by activating NF-kB, but its signaling pathway is poorly understood. Recent studies suggested PKCa and Raf-1 kinase might activate NF-kB during hypoxia, but the relationship between these events is not entirely clear. The objective of this paper is to determine whether human monocyte cell line (U937) exposed to hypoxia induce IL-1b and TNF-a expression involving PKCa/Raf-1/NF-kB pathway.

Methods U937 cells were subjected to 0, 1, 3, 6, 9 h of hypoxia (3% O 2 , 5% CO 2 , 92% N 2 ) with or without PKCa inhibitor, chelerythrine (10 mmol/L). PKCa and Raf-1 activity were analyzed by PKCa kit and Raf-1 kinase cascade assay kit. NF-kB binding activity was analyzed by electrophoretic mobility shift assay (EMSA). IL-1b and TNF-a mRNA levels were determined by RT-PCR. Results PKCa and Raf-1 activity increased to peak [(10.06 ± 1.83) mg/mg.pro, 4.97-fold, respectively] after 1 h of hypoxia and remained to 9 h [(5.55 + 1.16) mg/mg. pro, 3.19-fold, respectively], which were significantly higher than those of 0 h of hypoxia (P < 0.01). NF-kB binding activity significantly increased after 1, 3, 6, 9 h of hypoxia (78.5 ± 4.2 ~143.7 ± 7.3 versus 43.1 ± 3.8, P < 0.01). IL-1b mRNA and TNF-a mRNA significantly elevated after 3 h of hypoxia (2.37-fold, 2.96-fold, respectively, P < 0.01) and remained to 9 h. Conversely, inhibiting PKCa with chelerythrine abolished the positive effects of hypoxia on PKCa, Raf-1 and NF-kB activation. IL-1b and TNF-a mRNA did not significantly increase after 1, 3, 6, 9 h of hypoxia with chelerythrine (P > 0.05).

Conclusion These results indicate that U937 cells under hypoxia induce IL-1b and TNF-a expression and this induction requires PKCa activation, and PKCa-mediated Raf-1/NF-kB pathway. Furthermore, PKCa inhibitor, chelerythrine can block this signaling pathway and abolish the induction of IL-1b and TNF-a by hypoxia. Our data suggest that the induction of IL-1b and TNF-a in U937 cells by hypoxia involving PKCa/Raf-1/NF-kB pathway. Purpose To give a way to solve one problem in IL1610 Blood Gas Analyzer. Problem Blood sample can not be injected or aspirated into housing of the solution, and the analyzer show the signal of 'Er'. Results First, flush liquid could not be aspirated in to housing of the solution; second, calibration 1 and 2 could be carried out and waste liquid went into waste bottle; third, there was slight abnormal second when the pump winding, we found that it was the aging peristaltic pump tubes. After changing tube, the analyzer work normally.

Conclusion Sometime, small error can influence the diagnosing and treatment of patients. Troubleshooting for faults and the solving of more simple problem may be carried out directly by the operator. When we are familiar with the character of this blood gas analyzer, we can offer more exactitude and quickly value for clinical and laboratory work. Background Gastro-oesophageal reflux is a common clinical disorder associated with a variety of respiratory symptoms, such as chronic cough and asthma, but the mechanisms is still not clear.

The microvascular leakage was induced by intraesophageal 1N HCl in the presence or absence of neutral endopeptidase inhibitor phosphoramidon and NK1-receptor antagonist FK888 or SR140333 in anesthetized guinea pigs. The airway plasma leakage was evaluated by measuring extravasated Evans blue dye in the animals pretreated with propranolol and atropine. Data are reported as mean ± SD. Background Eosinophilic bronchitis (EB) presents with chronic cough and sputum eosinophilia, but without the abnormalities of airway function seen in asthma. The difference in the characteristic of airway inflammation between EB and asthma is uncertain. This study is to explore the pathological feature of airway in EB and compared with those with cough variant asthma (CVA) and asthma. Methods Flexible fibre optic bronchoscopy was performed in 11 patients with EB, by contrast with 10 patients with CVA, 14 patients with asthma and 10 healthy controls. The mean thickness of the basement membrane was measured. Using immunohistochemical and special staining, the localization and density of inflammatory cells were detected in the submucosa of bronchial with EB and CVA patients.

The mean thickness of the basement membrane was significantly increased in the subjects with those with EB 2.92 mm (2.1-6.5), CVA 5.64 mm (3.23-8.48) and asthma 9.08 mm (6.61-11.99) than in the healthy controls 2.08 mm (1.62-3.40). There were also significant differences among the three groups. The number of mast cells and eosinophils in the submucosa of bronchial from subjects with EB [75/mm 2 (35-112), 7/mm 2 (0-31)] were substantially decreased than those in subjects with CVA [148/mm 2 (34-200), 114/mm 2 (1-768); P < 0.05].

Conclusion EB is an inflammatory disorder of the airways with the characteristics of various inflammation cells infiltration. The level of infiltration of inflammatory cells and the mean thickness of the basement membrane is less severe than that in CVA and asthma which may be one of the reason that why EB without airway hyperresponsiveness. 

Background Gastro-oesophageal reflux (GER) is an important etiological factor of chronic cough, although continuous 24-hour ambulatory esophageal pH monitoring is a golden standard for determining GER, which is usually expensive, intolerance and incompliance, and is still not widespread used. This study is to determine if the clinical feature is diagnostically useful for GER induced cough (GERC). Methods All patients were evaluated by a modified Irwin's diagnostic protocol and continuous 24-hour esophageal pH monitoring was performed in forty GERC suspect patients. The final diagnosis of GERC required having cough disappear or substantially improve with anti-reflux therapy. The feature of clinical manifestation was compared with non-GER induced cough (NGER).

Results The mean ± SD of age was 37.7 ± 13.9 years in GERC (n = 20) and 40.0 ± 13.0 years in NGER (n = 23), The median of cough history was 61 (range, 3 to 360) months in GERC and 26 (range, 2 to 480) months in NGER. Cough associated with meals were present in 13 out of 20 patients in GERC, significantly higher than NGER (2 out of 23 patients) (P < 0.01). The specificity, positive predictive value and the sensitivity of cough associated with meals for GERC were 91.3%, 86.7% and 65%, respectively. Regurgitate associated symptom were present in 11 out of 20 patients in GERC, which was no significantly difference with NGER (8 out of 23 patients). The specificity, positive predictive value and sensitivity of regurgitate associated symptom were 65.2%, 57.9% and 55%, respectively. Background Eosinophilic bronchitis (EB) presents with chronic cough and sputum eosinophilia, but without the abnormalities of airway function seen in asthma. The difference in the characteristic of airway inflammation between EB and asthma is uncertain. This study is to explore the exfoliative cytology feature of airway in EB by observed the defferential cell counts in sputum and bronchoalveolar lavage fluid (BALF) with EB patients and the results were compared with those with cough variant asthma (CVA) and asthma groups. Methods Induced sputum by hypertonic saline aerosol inhalation was performed in 49 patients with EB, 28 patients with CVA, 43 patients with asthma and 38 healthy controls. BAL were performed in 12 patients with EB, 10 patients with CVA and 4 patients with asthma. Differential cell counts were performed in sputum and BALF.

Results Percentage of eosinophils in sputum were significantly increased in the subjects with those with EB 5.0% (3-64), CVA 7.2% (0-65.5) and asthma 26.0% (2-88) than in the healthy controls 0.0% (0-2.5). There were significantly differences between the asthma and EB group (P < 0.05). Percentage of eosinophils in BALF were significantly decreased in the subjects with those with EB 0.0% (0-4.5) than in the CVA 5.3% (0-11) (P < 0.05) and asthma group 11% (7-13.8).

Conclusion The airway inflammation with eosinophils in EB is mainly localized in the central airway, but less milded in the peripheral airway may be closely related to the reason that why EB without airway hyperresponsiveness. Background Measurements of lung function in 5 year old children requires a different approach to that used in older children. Due to their state of physiological and emotional maturity they cannot reliably perform spirometry to ATS standards, with end-of-test criteria being the most difficult to satisfy. Objective The present study was conducted to determine appropriate lung function variables for use in 5 year old children.

Method Lung function measurements were attempted in 198 children attending the 5 year assessment of a longitudinal study investigating risk factors for the development of asthma. Children were assessed when free of current respiratory infections.

Results 37 children (18.7%) had current asthma. Lung function, including bronchodilator response was successfully measured by spirometry (Sensormedics Vmax29) in 110 children and by forced oscillation (2-48 Hz Chess Medical i2M) in 143. As the validity of FEV 1 has been questioned in this age group, we also investigated the use of FEV 0.5 as an outcome variable. While lung function tended to be lower in the asthmatic children, this only reached statistical significance for pre-bronchodilator FEV 0.5 /FVC (70% v 75%, P < 0.05). Asthmatics had great increases in lung function with bronchodilator (FEV 0.5 : 10.0% v 4.6%, FEV 0.5 /FVC: 6.2% v 2.8%, respiratory reactance at 6 Hz: 36.2% v 18.7%) than non-asthmatics. Conclusion This study demonstrates that useful measurements of lung function can be made in 5 year old children but standard adult criteria and outcome variables are not suitable for this age group. 

Background The purpose of this prospective study is to predict postoperative lung function and exercise capacity values including DLCO during exercise, and to evaluate the effect of lung resection on lung function and exercise capacity values including DLCO during exercise after one year. Methods Fifty-seven patients undergoing lung resection at Vancouver General Hospital from October 1998 to May 1999. Lung function including FEV1 and FVC, and exercise capacity including maximal oxygen uptake (VO2max/kg) and maximal workload (Wmax) were obtained by routine procedure. We used a modification of the single breath DLCO technique, the 3-equation method (3EQ-DLCO), to determine DLCO during rest and during steady state exercise at 70% of the maximal workload reached in a progressive exercise test, and the increase in 3EQ-DLCO during exercise, (70%-R) DLCO%, was determined. Calculation of the predicted postoperative (ppo) variables were performed using preoperative testing data and the extent of resected bronchopulmonary segments. All function variables were collected from 30 patients again after lung resection.

The ppo values from the calculation were lower than but not significantly different from actual postoperative values. Following lung resection, there was a more significant reduction in lung function including FEV1, FVC, and DLCO (12%, 13%, and 22%) than that in exercise capacity including VO2 max/kg and Wmax (2.1 ml/min/kg (or VO2 max 8%) and 12 watts (or 7%)). But the postoperative DLCO increase during exercise was not significantly decreased (2%).

Conclusions This study confirms that postoperative lung function were significantly decreased, but (70%-R) DLCO% was not significantly decreased. These findings suggested that exercise capacity, especially (70%-R) DLCO%, is more conservative and accurate for preoperative evaluation and postoperative prediction. Background Eosinophilic bronchitis (EB) is an important cause of chronic cough. Some reports had shown that the airway pathological features in EB are similar to asthma. This study is to observe the cough sensitivity in EB, cough variant asthma (CVA) and classic asthma subjects. Methods Capsaicin challenge tests for measuring cough sensitivity were performed on 10 healthy volunteers, 8 subjects with EB, 9 patients with CVA and 7 patients with classic asthma. Each subject inhaled progressively doubling concentrations of capsaicin (0.98 ~1000 mmol/L) by a breathactivated dosimeter until the concentration inducing five or more coughs (C5) was reached. The LogC5 was calculated as the cough reflex sensitivity.

The mean value of LogC5 in normal subjects was 2.61 ± 0.40. The mean LogC5 value in subjects with EB (1.61 ± 0.77) was significantly lower than normal subjects (P < 0.01), CVA (2.26 ± 0.59, P < 0.05) and classic asthma (2.14 ± 0.43, P = 0.01). There was no significant difference between normal subjects, CVA and classic asthma patients (P > 0.05).

Conclusion In contrast to CVA and asthma, the increased cough sensitivity is a physiological characteristic of EB. The mechanisms of EB may be different from asthma. Venous thromboembolism is a common and under diagnosed clinical problem amongst hospitalized patients. Venous thromboembolism results in significant morbidity and mortality. We carried out this study to evaluate the demographics of venous thromboembolism at our tertiary care hospital in Pakistan. We conducted a retrospective chart review of all patients diagnosed with venous thromboembolism (deep venous thrombosis and or pulmonary embolism) for the period 2000 and 2003.

One hundred and nine patients were diagnosed with deep venous thrombosis and or pulmonary embolism during the study period. Mean age was 55.7 (17.9) years. Males accounted for 47% of cases and females 53%. The majority of diagnosed cases (74%) were from medical and 26% from surgical services. Immobilization was the most commonly identified predisposing factor in 46% cases, followed by malignancy in 20%, postoperative status in 20% and 12% with hypercoagulability (previous venous thromboembolism, anticardiolipin antibody syndrome and oral contraceptive use). Only 40% of patients were on any form of prophylaxis; the most commonly used method was subcutaneous heparin (58%) with compression stockings and low molecular weight heparin in 21% each. The d-Dimer test and venous doppler scan were positive in 56% and 60% of patients respectively. Spiral CT was diagnostic in 82% cases. Prophylaxis against venous thromboembolism is significantly underutilized at our hospital. Spiral CT was confirmed as the best diagnostic modality. Serum D-dimer assay has emerged as a very popular test for screening patients with acute venous thromboembolism. Most studies quote a sensitivity upto 90%. The objective of our study was to evaluate the clinical utility of D-dimer assay in our local setting. We conducted a retrospective chart review of all patients diagnosed with acute venous thromboembolism (deep venous thrombosis and or pulmonary thromboembolism) for the period 2000 to 2004. The diagnosis was confirmed by Spiral CT scan, venous Doppler, Perfusion scan or Pulmonary angiogram. All patients' who also had a D-dimer assay were included in the study. In our hospital, D-dimers are detected using a 'D-dimer Plus' kit. This relies on a latex turbidimetric measurement of the concentration of D-dimer in venous blood. The manufacturer claims a correlation coefficient of 0.99 with a y-intercept of -109 and a slope of 1.21 when compared to commercially available assays. One hundred and fifty nine patients were diagnosed with deep venous thrombosis and or pulmonary embolism during the study period. D-dimer measurements were done in 114 (72%) patients. Seventy-two patients had a positive result whilst forty-two were negative. The calculated Sensitivity of the test comes to 63%. The sensitivity of the test in our institution was too low to condone its use as a screening tool for acute venous thromboembolism.

After exposure to ozone, humans develop neutrophil infiltration of the nasal and bronchial mucosa. To investigate the events contributing to inflammatory cell recruitment in the bronchial mucosa we exposed 10 healthy nonsmoking volunteers to 400 ppb ozone or filtered air for 2 h at rest on two separate occasions. Bronchial biopsies were performed 6 h after ozone/filtered air exposure. The biopsies were embedded in glycol mathacrylate and immunostained for inflammatory cells, including neutrophils, mast cells, total T-cells (CD3), T-cell subsets CD8 and CD4, macrophages, eosinophils adhesion molecules (P-selectin, E-selectin, ICAM-1, VCAM-1) cytokines (TNF-a, IL-1b, GM-CSF, IL-6), chemokines (IL-8 and RANTES), and nuclear factor NF-kB. No significant changes were seen in the number of T-cells and T-cell subsets, macrophages, eosinophils or percentages of vessels expressing P-selectin, VCAM-1, GM-CSF, IL-6 and RANTES in the biopsies. The number of neutrophils and mast cells in the submucosa was significantly higher after ozone exposure (P = 0.009 and P = 0.005 respectively). The percentage of vessels expressing E-selectin (P = 0.01), ICAM-1 (P = 0.005), IL-8 (P = 0.02), TNF-a (P = 0.02), IL-1b (P = 0.009), and NF-kB (P = 0.05) increased significantly after ozone exposure versus filtered air exposure. Exposure of normal subjects to ozone increases the expression of proinflammatory cytokines resulting in upregulation of IL-8 and adhesion molecules via activation of NF-kB, leading to neutrophil infiltration in the bronchial mucosa. Purpose Ciliary beat is a key determinant of mucociliary clearance mechanism, which maintains the sterility of the lower respiratory tract thereby the respiratory health against inhaled pathogens and accumulating debris in the airways. Gelomyrtol Forte (GF) a phytomedicine claimed to exhibit seretomucolytic functions and anecdotally used to as treatment for COPD and sinusitis. Nonetheless, the effects of GF on ciliary beat have not been studied in a systematic manner. Methods We have examined the effects of GF on the ciliary beat frequency of human respiratory mucosa. The latter was obtained without anaesthetics from the inferior turbinate of 13 healthy subjects, and suspended in M199 which contained 0, 100 ng/ml and 250 ng/ml of GF. Ciliary beat frequency (CBF) for each GF concentration was examined using a phase contrast microscope and a photometric system linked to a digital converter as per standard established protocol in our laboratory at 0 and 4 h.

The following table shows our preliminary data:

Mean ± SD CBF (Hz) GF level (ng/ml) 0 h 4 h 0 1 1.25 ± 1.5 11.66 ± 1.9 100

11.54 ± 1.5 12.31 ± 1.6 250 11.86 ± 1.8 12.67 ± 1.7

Paired Samples t-test between 0hr and 4hr on all groups showED significant increase in CBF after incubation (0 ng/ml GF, p = 0.01; 100 ng/ml, p < 0.001; and 250 ng/ml, p < 0.001). There was a trend towards higher CBF after GF incubation at 4 h. However, One-way Analysis of Variance showed no significant difference at 4 h between CBF obtained with 0, 100 or 250 ng/ml. Background Though CPAP can improve anoxemia has been approved, Results of the cardiovascular effects of CPAP in ACPE have been controversial, with increased, decreased, or unchanged cardiac output (CO) when CPAP applied. Methods ACPE happened when healthy dogs elevated blood volume significantly. Cardiac output (CO), heart rate (HR), blood pressure (BP), pulmonary arterial pressure (PAP), pulmonary artery wedge pressure (PAWP), central venous pressure (CVP) and intrathoracic pressure (Pt) were measured and recorded before model making and at spontaneous breath, 5 cmH 2 O, 10 cmH 2 O, 15 cmH 2 O CPAP successively in ACPE dogs. Results ACPE occurrence induced significant reduction of CO, BP (P < 0.05) and significant elevation of PAP, PAWP, HR, and Pt (from -(4.90 ± 0.09) to -(10.90 ± 0.75) cmH 2 O (P < 0.05)). PaO 2 decreased (P < 0.05). CPAP at 5 cmH 2 O induced a proper increase in CO and pt (P < 0.05), a slightly drop in PAP, PAWP and HR (P < 0.05). Anoxemia was corrected. CPAP at 10 cmH 2 O decreased CO (P < 0.05), elevated PAP and PAWP (P < 0.05) and Pt (P < 0.01). HR and BP remain. CPAP at 15 cmH 2 O decreased CO to 0.82 ± 0.07 (P < 0.05), elevated PAP and PAWP (P < 0.01), Pt was -(0.82 ± 0.37) cmH 2 O. Conclusions Proper CPAP can increase CO by regulating Pt, and then increase HR, BP, PAP and PAWP; while more than 10 cmH 2 O CPAP will depress cardiac function.

Objective To investigate the change and clinical significance of nitric oxide (NO) endothelin-1 (ET-1) in plasma during acute exacerbation of chronic cor pulmonale before and after treatment.

Methods The plasma concentration of NO and ET1 -were detected in 63 patients with acute exacerbation of chronic cor pulmonale during mild attack or moderate to severe attack by using radioimmunoassay, and their correlation were observed. Resutls The level of plasma NO, ET1-1 were significantly increased in patients with acute exacerbation of chronic cor pulmonale as compared with controls group. The difference was statistically signiticant (The group of mild attack: P < 0.5, the group of moderate and severe attack: P < 0.01, P < 0.01. The level of plasma NO, ET-1 were significantly decreased in patients with moderate to severe attack after treatment than that before treatment (P < 0.01). The NO and ET-1 were positively correlated with acute exacerbation of chronic cor pulmonale. Conclusion The increased level of plasma NO and ET-1 play vital role in the pathophysiologic prosess of chronic cor pulmonale, And may be involved in the pathophysiologic process of chronic cor pulmonale, and may be involved in the development of pulmonary hypertension. Objective To investigate the role of modulating immune response of mice with rBCG vaccination expressing foreign antigen Der p2 in the form of lipoprotein.

Methods Two groups of seven 6-8 weeks old and two groups of seven newborn BALB/c mice were vaccined intraperitoneally with 100 mL of 10 6 CFU rBCG or BCG respectively. The control group was injected with 100 mL saline. Six weeks later, all animals were injected with Der p2 (20 mg). Further two weeks later, the concentrations of IL-4 and IFN-g from both immunized mouse serum and splenocyte culture supernatant (STLCS) were determined by ELISA, and Th subset were determined by double fluorescent staining and flow cytometry. Results After vaccine, the serum and STLCS from both rBCG and BCGimmunized group of adult or newborn mice had significantly higher level of IFN-g and lower level of IL-4 than that from control groups. Moreover, the level of IFN-g in STLCS from rBCG-immunized was significantly higher than that from BCG-immunized mice. Besides, there was the larger percentage of CD4+/IFN-g+ cells in spleen from rBCG and BCG-vaccined mice than that from control group. But the results of the percentage of CD4+/IL-4+ cells is reversed. The percentage of CD4+/IFN-g+ cells in spleen from rBCGvaccined mice was larger than that from BCG-vaccined group.

Conclusion Both rBCG and BCG could stimulate Th1 predominant immune response, when injected intraperitoneally into adult or newborn BABL/c mice. The rBCG-expressing antigens can be used as the antigenspecific vaccines against allergic diseases by regulating the balance of Th1/Th2. Background Single lung transplantation (SLT) and bilateral lung transplantation (BLT) are routinely performed in patients with COPD and primary pulmonary hypertension (PPH) and secondary pulmonary hypertension (SPH) in western country. But in most Asian country it is far from routine work. Especially for late COPD, we usually treat the late COPD by both lung volume reduction (LVRD) and single lung transplantation. We have more 120 cases LVRD and 4 cases SLT experiences. We reviewed our 4 cases experience with lung transplants for COPD and SPH to determine if any advantage exists with SLT. Methods For 3 COPD cases, ones Fev1 was 23%, the others were respiratory failure, could not accepted the lung function test; the SPH fev1 was 30%, but O2 was less 9 kPa. Two cases HLA and height were match, gene match point were over 6 and the other were unmatch. The 2 COPD and one SPH cases accepted Right SLT, 1 COPD case accepted left SLT. A small muscle spare lateral thoracotomy incision is made for SLT. With ventilation to the contralateral lung, the standard pneumonectomy is being perfomed. During the implantation bronchial anastomosis is performed as the first one. Pulmonary artery anastomosis was followed. The procedure continues with anastomosis of pulmonary veins. De-airing of venous system follows. Venous system was tighten by pulling the ends of the above mentioned venous suture, which was still not tied. Microorganisms from the bronchial secretion of the donor and the recipient are being examined routinely. Immunosuppressive therapy was introduced immediately after transplantation. FK 506 was administered in dose 0.2 mg/kg/day by oral. Later the dose was adopted according to the blood level about 15-20 ng/ml at first week, 10-15 ng/ml at second week and 10-12 ng/ml at following week. MMF was given 0.5 mg B.i.d orally if white cell was over 6000. Single intravenous dose of Methylprednisolone was applied (500.1000 mg/ according to the patients weight) just before reperfusion. Further dosage was within 1 mg/kg/day postoperatively. The immunosuppressivum were Anti-thymocite globulin (ATG) and anti-T-cell-body. Its dosage was variable. It was applied as an induction therapy only during the early post-transplant period. 50 mg daily dosage ATG was given continuously 7 days. The two cases who HLA are match have not given steroid after SLT 6 months. Now usually they just take once FK 506, MMF one day and are checked their FK 506 blood level and follow up their life situation in out patient department every 3 month after SLT. Results All cases have been alive after SLT. The average hospitalization day after operation were 28 days, and the blood lost average each case was 1500 ml. In one case acute rejection was occurred in 5 th day, and the other was occurred in 11 th month, both of them were healed by induce and steroid treatment. The longest case is being 20 month. The first 2 cases who had SLT in 2003, they have being work their former jobs after they accepted SPT 4 months. The average fees were nearly 50000 USD. The other two cases were too early to work. The FEV1 each was 47%, 45% in one year. The 6 minute-walk distance each is 650 m, 450 m, and 420 m at the 40th days after SLT. Conclusion We believe that SLT is the last and effective procedure of choice for COPD. It can improve the Patients pulmonary function indeed and make them enjoy their life as well as working ability as before again. Objective Ozone is a broad-spectrum germicide. It could kill bacteria breed, sporangium, virus, fungi etc. and were widely applied in hospital sterilize, especially in sewerage and room air. For the physical characteristic of ozone, it were hard to reserved and must be used as soon as possible while were produced, as we know the ozone generator need 220 voltage in traditional, limited widely applying in the movement public environment, for example the obturator vehicle. This study focus on the room air purify in hospital environment with low voltage ozone generator. Methods 12 voltage ozone generator provided by patentee professor Kong xiangong. Fifty liters per hour of exhaust volume with 5A electric current could produce 3.5 gram per hour ozone, go on the room air sterilize in hospital obturator room which the acreage was 38 square meter, we enactment 10 minutes sterilize and rest 5 minutes, before and after the sterilized we cultivate the room air with 6 grade sieve pore air bounce method, synchronous check room air ozone content, temperature, humidity.

The average room air cultivated bacteria number were 455.1 ± 204.4 cfu/m 3 , with ozone concentration was 0 ppm, after sterilized the average room air cultivated bacteria number were 69.7 ± 64.6 cfu/m 3 , with ozone concentration 7 ppm, the die out ratio was 84.7%, after sterilized the average room air cultivated bacteria number less than 157 cfu/m 3 without exception, and further more 33% of the die out ratio achieve 100%, according to the sanitation standard of hospital sterilize number GB15982-1995, it could match the II grade environment standard, such as ordinary operation room, delivery room, baby room, premature room, accommodate room, asepsis room, burn department, ICU etc; at the same time we compared the results with the low ozone ultraviolet radiation light, this method widely used in clinical which power 30 watt continued 30 minutes during 3/8 to 28/8, it showed no significant different between two group results (P > 0.05). Conclusion This low voltage ozone generator had effective to sterilize the obturator room air and could match the the second grade environment in hospital standard. its efficacy also matched the low ozone ultraviolet radiation light in clinical used on traditional which only needed half time. Background Sepsis remains a major cause of morbidity and morality in hospitalized patients.The primary cause of sepsis result from activation of host effector cells, by endotoxin, the lipopolysacchride (LPS) associated with cell membranes of Gram-negative bacteria. Whereas the acute-phase rise lipopolysacchride-binding protein (LBP) initate and amplify the process. Nowadays more and more studies show that blockade the initial process can prevent the development of sepsis. Methods Select peptide which block LBP inflammatory function site by phage display, then examined the effects of the peptide on the binding of FITC-LPS to Phorbolmyristate-acetate (PMA) differentiated U937 (human macrophage-like cell line) by flow cytometer, the mRNA expression and protein production of TLR4 and CD14 by RT-PCR and Western blot, the NFk B DNA-binding activity by EMSA, the TNF a (tumor necrosis factor alpha) production by ELISA.

The LBP inhibitory peptide with the sequence of WKXRKXFXKXXG is homologus with 91-102 aa WKVRKSFFKLQG of LBP which could be inflammatory function site.The binding of LPS and LBP inhibitory peptide inhibited the binding of LPS to PMA differentiated U937, reduced the mRNA expression and protein production of CD14 and TLR4, suppressed the NFk B DNA-binding activity and therefore reduced the production of TNF a. Conclusion Our results suggest that LBP inhibitory peptide might ameliorate sepsis through blockade the binding of LPS to U937, suppression of NFk B DNA-binding activity and reducing cytokine production. Background Warfarin is effective in preventing recurrent venous thromboembolism but is also associated with a risk of bleeding. In this study, the warferin initiation and maintenance nomograms for PTE patients under different clinic setting were observed. Methods 79 patients with confirmed acute PTE were assigned to accompanying with chronic diseases or organ dysfunction (the first group), or accompanying with a chronic disease but without organs dysfunction (the second group), or no any chronic diseases (the third group). Then the subgroups were assigned by the initial warfarine to either 2.5 mg or 5 mg. The initiated doses, the time to achieve a therapeutic international normalized ratio (INR), the proportion of patients whose INRs exceeded the therapeutic range, and the maintenance doses were analyzed prospectively.

Results By using 2.5 mg initial warfarine, the proportion of patients achieved a therapeutic INR by the 5th day of treatment were 82.35%, 42.83% and 0 in the first, second and third groups. The time to achieve a therapeutic INR in the third group was longer than the first one, but was no difference compared to the second group. By using 5 mg warfarine, the proportion of patients achieved a therapeutic INR were all 100% among three groups, and a therapeutic INR was all exceeded in the first group. Maintaining and total doses needed to be achieved therapeutic INRs were lower in the first group than either second or third group. The proportion of patients whose INRs exceeded the therapeutic range was higher in the second than the third groups. Conclusions 1) in the patients accompanying with chronic diseases or organs dysfunction, the initial and maintaining dosage of warfarine with ~2.5 mg and ~2 mg could be more safe. 2) in the patients accompanying with a chronic disease without organs dysfunction, the initial dosage with 5 mg could be chosen but reduced afterwards; 3) the initial and maintain dosage with ~5 mg and ~3.75 mg could be suitable in the patients no accompanying with any chronic disease. Background Hypoxia may enhance the generation of free radicals in skeletal muscle, which may in turn affect contractility. In this study, we determined the effect of hypoxia on diaphragm muscle peroxynitrite (ONOO) formation under resting conditions and after fatiguing contractions. Methods Rat diaphragm muscle strips were dissected and mounted in tissue baths under hypoxic (PO 2~ 6.5 kPa) or hyperoxic (PO 2~ 88 kPa) conditions (60 cin). Force frequency relationship was determined. Subsequently, muscle nitrotyrosine formation was assessed as a measure of ONOO formation. In addition, the effects of fatiguing isotonic contractions were determined on nitrotyrosine formation under hypoxic and hyperoxic conditions (60 min). Results Hypoxia depressed force frequency relationship (P < 0.001 vs. hyperoxia). Nitrotyrosine optical density of protein bands already present under hyperoxic conditions was significantly enhanced by hypoxia. Also, additional protein bands were detected during hypoxia at 295 and 243 kDa. Fatigue endurance was reduced under hypoxic conditions (P < 0.05 vs. hyperoxia). Fatiguing contractions increased nitrotyrosine formation under hyperoxic conditions (P < 0.05 vs. rest), but not under hypoxic conditions (P > 0.05 vs. rest).

Conclusions These data are the first to show that hypoxia is associated with increased skeletal muscle nitrotyrosine formation. In addition, strenuous work increases ONOO formation in rat diaphragm muscle. This abstract is funded by: Netherlands University Fund for International Cooperation

A population survey of respiratory symptoms in the elderly

Background Adiponectin (AN), one of adipocyte-secreted protein, has a unique feature about fat metabolism and body weight. It is well known that the blood levels of AN in healthy subjects are higher than those in obese subjects. Weight loss in patients with chronic respiratory failure (CRF) is a major complication, and is an important determinant of the survival prognosis. Therefore, we hypothesized that the blood levels of AN might be further higher than in healthy subjects, and that it might play a role for body weight loss. Methods In 11 subjects with CRF (male: female = 3 : 8, Age: 52.8 ± 9.7 years). The body composition analysis using bioelectrical impedance analysis (In Body®), biochemical measurements of the blood, and pulmonary function tests were performed. AN was assayed by validated sandwich ELISA using an adiponectin-specific monoclonal and polyclonal antibody. Insulin resistance was calculated by the homeostasis assessment (HOMA-R). All the subjects were clinically stable at the time point of those measurements. Results AN concentration is negatively correlated with body mass index (BMI) and fat mass (P < 0.05), but not with body fat mass, insulin resistance, or any parameters of lung functions. The patients with extremely high AN level had low levels of BMI. Conclusion Adiponectin is a candidate for one of major contributors of weight loss in patients with chronic respiratory failure. Background Pulmonary alveolar microlithiasis (PAM) is a rare disease characterized by widespread laminated calcispherites in alveolar spaces in the absence of any known disorder of calcium metabolism. Although the pathogenesis is unknown, one possibility is that it is an inborn error of metabolism, and thus is regarded as a hereditary lung disease. In presumption of autosomal recessive trait we are trying to clone the responsible gene for the disease. Method Genomic DNA from patients who suffered from PAM and consanguineous marriage in their family tree were analyzed by SNP chips obtained from Affymetrix.

GeneChip analysis of the first patient showed stretches of homozygous SNP sites that covers 1/8 of whole genome when added up. These homozygous stretches are generated as a result of consanguineous marriage, and are considered to candidate regions that harbor the responsible gene for PAM. We are analyzing the 2nd and the 3rd case by the GeneChip. By combining the results, the length of the candidate regions could be shortened geometrically, and 3 cases will be enough for picking up less that 100 genes that will be thoroughly searched for their mutations. Background Respiratory diseases may cause considerable disability in the elderly because of their limited respiratory reserve related to ageing. This study aims to study the prevalence of respiratory symptoms and diseases in the elderly Chinese in Hong Kong. Methods A telephone survey was conducted by random household telephone interview from November to December, 2003. Subjects with age ≥70 years were asked to complete a respiratory questionnaire 1 . Subjects were subsequently invited to undergo lung function (spirometry, bronchial challenge) and skin prick tests. Results 1522 subjects (697 male and 823 female) completed the telephone interview. The mean age was 76.4 ± 5.6 years old with 71.1%, 17.2% and 11.7% being non-smokers, ex-smokers and current smokers respectively. At lease one respiratory symptom was identified in 673 (44.2%) subjects. The most frequently reported symptoms were morning phlegm (33.9%), breathless at rest during the day (12.3%) and wheeze over the past 12 months (12.2%). Of the self-reported respiratory diseases, 2.8%, 5.7%, 7.7% and 3.1% had history of tuberculosis, asthma, chronic bronchitis and emphysema respectively. Of the 174 subjects with asthma, chronic bronchitis or emphysema, 96 (55.2%) had sought medical advice in the past 12 months. Among the 153 subjects who had undergone lung function testing, 17 and 13 subjects had FEV1 <80% and <70% predicted respectively. Positive bronchial challenge and skin prick test were identified in 14.3% and 19.6% of subjects respectively. Conclusion Respiratory symptoms, chronic respiratory diseases, bronchial hyperresponsiveness and skin atopy were common among Hong Kong elderlies. (This study is supported by the Hong Kong Lung Foundation Grant.)

Objective To investigate the expression and activation of mitogenactivated protein kinase (MAPK) and the relation with inflammation mediator production in the lung of rat with acute lung injury (ALI), and further to explore the role for MAPK signal transduction system in ALI. Method The murine model of ALI was estabalished by the method of 'two hit', instillation of little acid and subsequent lipopolysaccharide into lungs. Immunohistochemical techniques was uesd to detect the expressions of Raf-1, MEK-1, Extracellular signal-regulated kinase (ERK), phosphorylated-ERK (P-ERK) and iNOS, TNF-a, and compare them with control group. Results In the experimental rat lungs, MEK-1, ERK and P-ERK were intensively expressed, but Raf-1 expression was relatively weak. In addition to their expression, the stronger expression of iNOS and TNF-a were seen in the same lung area. Nevertheless, only the minimal exprssions of these kinases were detected in normal control lungs and few cells showed immunoreactivity staining, especially Raf-1 and P-ERK. The control lung expressed lower levels of iNOS and TNF-a than ALI. Significant differences were noted in the experimental and control groups (P < 0.05). Conclusion Raf-1 to MEK-1 to MAPK signal transduction system were activated in the lungs of ALI rat, with the elevated expression of inflammatory mediator NO, TNF-a. The activation of Raf-1 to MEK1-1 to MAPK could lead to the activation of polymorphonuclear neutrophile and macrophage which synthesised and secreted NO, TNF-a.

Objective This study used a model of pulmonary thromboembolism (PTE) in rats induced by injection of thrombus to investigate the mechanism of the pulmonary vascular remodeling. Methods Male SD rats were divided averagely into PTE and sham group. PTE model was induced by injection of thrombus, and injected subcutaneouly with Tranexamic acid. Shams were no thrombus. Rats were sacrificed at the 1st, 3rd, 7th and 14th days (d). The lungs was stained by HE, Masson and PTAH. The IH stainings of MMP-9, TIMP-1 and PDGF-BB were taken. The hybridization in situ stainings of MMP-9 and TIMP-1 were made. Results Inner elastic membrane (IEM) in embolic vessel wall was destroyed and the SMCs migrated into subintimal area. The intima thicken and neointima appeared (pulmonary vascular remodeling). MMP-9, TIMP-1 and PDGF-BB were expressed in the SMCs, endothelial cells of embolic vessel and neointima in the PTEs. The expressions reached its culmination at the 3rd and 7th d respectively and became weak at the 14th d. The expression of a-SMA was seen in the cells of the neointima of the embolic vessel. Conclusion PTE could lead to the hyperplasia of endothelial cells, the destruction of IEM. SMCs migrate into the subintimal area and cause the occurrence of neointima and vascular remodeling. The lopsided development of MMP-9/TIMP-1 involve in the pulmonary vascular remodeling associated with PTE.