Bcl2L12 Contributes to Th2-Biased Inflammation in the Intestinal Mucosa by Regulating CD4+ T Cell Activities.

The Th2-biased inflammation and immune deregulation play a critical role in the pathogenesis of ulcerative colitis (UC). Recent studies indicate that the Bcl2-like protein 12 (Bcl2L12) is associated with immune deregulation of UC. This study aims to investigate the role of Bcl2L12 in the induction of aberrant Th2-biased inflammation. In this study, peripheral blood samples were collected from patients with inflammatory bowel disease. The Th2 cell activities were analyzed by flow cytometry, real-time quantitative RT-PCR, and Western blotting. Mice with Bcl2L12-knockout CD4+ T cells were used in the experiments. The results showed that the expression of Bcl2L12 was detected in peripheral CD4+ T cells, which was significantly higher in UC patients than in healthy subjects. A positive correlation between the expression of Bcl2L12 and Th2 cytokines was detected in CD4+ T cells from UC patients. Naive CD4+ T cells with Bcl2L12 overexpression were prone to differentiate into Th2 cells. Mice with Bcl2L12 deficiency failed to induce the Th2-biased inflammation in the intestine. Bcl2L12 bound GATA3 to form a complex to enhance the binding between GATA3 and the Il4 promoter to enhance the expression of IL-4 in CD4+ T cells. CD4+ T cells with Bcl2L12 overexpression were resistant to apoptosis. In conclusion, the Bcl2L12 is a critical factor in the induction of aberrant Th2 polarization by upregulating Th2 responses and downregulating Th2 cell apoptosis. Bcl2L12 may be a novel therapeutic target in the management of the disorders with Th2-biased inflammation.

Clinical characteristics of tobacco smoke-induced versus biomass fuel-induced chronic obstructive pulmonary disease.

OBJECTIVE: To investigate differences in clinical features between tobacco smoke-induced and biomass fuel-induced chronic obstructive pulmonary disease (COPD). METHODS: We retrospectively analyzed 206 patients with COPD caused by exposure to tobacco smoke and 81 cases of COPD caused by exposure to biomass fuels who received treatment in our hospital between 2011 March and 2014 March. Difference in general health status, clinical symptoms, the dyspnea score, and comorbidities between the two groups were compared. In addition, pulmonary function, grading, and acute exacerbations were also compared. RESULTS: (1) Difference in general health status: Male and female patients with COPD caused by exposure to tobacco smoke were 83.5 and 16.5%, respectively. Male and female patients with COPD caused by exposure to smoke from biomass fuels were 14.8 and 85.2% (χ2 = 27.2, P < 0.05), respectively. Tobacco smoke-induced COPD was more prevalent in men, and COPD caused by exposure to smoke from biomass fuels was more prevalent in women. After gender adjustment, body mass index (BMI) was lower in women with COPD caused by exposure to smoke from biomass fuels than those by tobacco smoke. There was no statistically significant difference in other indicators, such as age. (2): Difference in clinical symptoms: No statistically significant difference in the modified British Medical Research Counsel (mMRC) Questionnaire, a measure of breathlessness, was observed between the two groups. Dyspnea was more common in COPD patients that was caused by exposure to biomass fuels (38.3%) than by tobacco smoke (11.1%) (χ2 = 17.9, P < 0.05). The comorbidities of allergic diseases (such as allergic rhinitis, bronchial asthma) were more prevalent in COPD patients that was caused by exposure to smoke from biomass fuels (43.2%) than by tobacco smoke (18%) (χ2 = 16.1, P < 0.05). However, COPD comorbid with lung cancer was more prevalent in those cases that were caused by exposure to tobacco smoke (7.77%) than in cases caused by exposure to smoke from biomass fuels (3.7%) (χ2 = 9.7, P < 0.05). (3) Differences in grading of pulmonary function: After gender adjustment, patients with COPD caused by exposure to biomass fuels were mostly in grade B or D. (4) Exacerbations: No significant difference in exacerbations per year was noted between the two groups. CONCLUSIONS: Marked differences exist between patients with COPD caused by exposure to tobacco smoke and smoke from biomass fuels. Patients with COPD caused by exposure to biofuels are mostly females with lower BMI and often with many clinical symptoms and complications, such as allergic rhinitis and bronchial asthma. Such patients are often in stage B or D. Tobacco smoke-induced COPD is more prevalent in male patients, often with complications in the form of lung cancer.

[Wood smoke condensate had weak proliferation and strong necrotic effects on human airway smooth muscle cells].

OBJECTIVE: To investigate the effect of wood smoke condensate (WSC) on proliferation and necrosis of human airway smooth muscle cells (HASMCs). METHODS: Primary cultured HASMCs between passage 2 and 8 were divided into 3 groups: a control group, a WSC group and a cigarette smoke condensate (CSC) group. The viability of cells was examined by the CCK8 assays. The ratio of cellular proliferative stage (S phase) and cell cycle index were examined by fluorescent-labeled thymidine analogue uptake assays and flow cytometry. The expression of cyclin D1 was detected by quantitative reverse transcriptase polymerase chain reaction (Q-PCR) and Western blot. Cell apoptosis and necrosis were observed by the annexin-V and PI staining. Statistical analysis was performed by using the One-way ANOVA and LSD-t test. RESULTS: Cell viability reached peak at WSC 1 mg/L[(126 ± 12)%] and at CSC 10 mg/L exposure level [(142 ± 11) %] respectively. While at WSC 10 mg/L and CSC 60 mg/L exposure levels, cell viability decreased significantly to 86% and 76%, respectively, as compared with that of the blank control group[(100 ± 0)%] (q = 3.63- 9.33, P < 0.05). In the WSC 1 mg/L group, the cell proliferation ratio and the expression of cyclin D1 protein were (124 ± 20)% and 1.31 ± 0.64, respectively, the differences being significant as compared with the blank control group [(100 ± 0)%, 1.0 ± 0.0] (q = 5.85, 5.91, P < 0.05), while the expression of cyclin D1 mRNA and the percentage of S+G2M phase were 1.18 ± 0.21 and (103 ± 4)%, respectively, not significantly different as compared to the control group [(100 ± 0)%, 1.0 ± 0.0], (q = 1.16, 2.05, P > 0.05). In the CSC 10 mg/L group, the above-mentioned values were (204 ± 45)%, 1.80 ± 0.25, (140 ± 6)%, 1.48 ± 0.2, respectively, which were significantly higher than those in the blank control group (q = 5.38-16.51, P < 0.05) and in the WSC group (q = 3.33-15.35, P < 0.05). However, when HASMCs were exposed to WSC 10 mg/L, the cell death ratio was (13.39 ± 0.15)%, higher than that of the blank control group [(1.57 ± 0.41)%] and the CSC group [(6.61 ± 1.91)%] (q = 18.03, 10.34, P < 0.05). Apoptosis ratio in the CSC 40 mg/L group was [(61.8 ± 10.6)%], higher than that of the blank control group [(0.0 ± 0.0)%] and the WSC group [(1.7 ± 0.4)%] (q = 17.44, 16.95, P < 0.05). CONCLUSIONS: Exposure to WSC caused a weak proliferation of HASMCs, but resulted in cell necrosis instead of apoptosis at high doses. There was a slight difference in cell effects between the WSC group and the CSC group.

[The prevalence and risk factors of bronchiectasis in residents aged 40 years old and above in seven cities in China].

OBJECTIVE: To investigate the prevalence and risk factors of bronchiectasis in urban city of China. METHODS: A cross-sectional survey was conducted in 17 urban areas in Beijing, Shanghai, Tianjin, Chongqing cities, and Guangdong, Liaoning, Shanxi provinces. In this study, urban population-based cluster samples were randomly selected from each city/province. In the selected city communities, all residents at least 40 years old were recruited, interviewed with questionnaires and tested with spirometry. Each participant was asked whether he/she was ever diagnosed as bronchiectasis by physician, whether had symptoms of respiratory diseases and possible risk factors, etc. RESULT: Data of 10 811 participants was enrolled for analysis, with a response rate of 75.4% (10 811/14 337). The overall prevalence of physician-diagnosed bronchiectasis was 1.2% (135/10 811), with 1.5% (65/4382) in male and 1.1% (70/6429) in female, without statistical difference in gender (χ² = 3.289, P = 0.070). Prevalence of bronchiectasis increased with age (χ² = 31.029, P < 0.001). There were no statistical significances in crude prevalences of bronchiectasis among cities (χ² = 10.572, P = 0.103), while there was a significant difference among cities after adjustment with confounders (Wald value = 22.116, P = 0.001), by using logistic regression analysis. Logistic regression analysis showed, bronchiectasis was significantly associated with elder ( ≥ 70 years vs 40-49 years; OR = 4.11, 95% CI 2.29-7.36), the family history of respiratory diseases (having two subjects with respiratory diseases in family vs no suffered relatives; OR = 2.04, 95% CI 1.06-3.94), respiratory infection during childhood (suffering two kinds of respiratory diseases vs never; OR = 4.89, 95% CI 2.03-11.81), exposure to coal (OR = 2.30, 95% CI 1.17-4.52), chronic pharyngitis (OR = 3.96, 95% CI 1.38-11.40) and pulmonary tuberculosis (OR = 3.07, 95% CI 1.89-4.98), heart diseases (OR = 1.64, 95% CI 1.11-2.42) and lung cancer(OR = 18.61, 95% CI 7.67-45.18). CONCLUSION: The prevalence of bronchiectasis in population aged 40 years old and above in urban area in China is high and associated with multiple factors such as age, family history of respiratory diseases, respiratory infection during childhood, exposure to coal, chronic pharyngitis, pulmonary tuberculosis, heart diseases, lung cancer and so on.

Human mesenchymal stem cells elevate CD4+CD25+CD127low/- regulatory T cells of asthmatic patients via heme oxygenase-1.

Up-regulation of CD4+CD25+CD127low/- regulatory T cells (Tregs) is a new target in the treatment of asthma. Human bone marrow mesenchymal stem cells can up-regulate CD4+CD25+CD127low/- regulatory T cells in vitro, meanwhile, heme oxygenase-1 (HO-1) plays an important role in the development and maintenance of CD4+CD25+ regulatory T cells. However the mechanism has not yet been adequately understood. Hence, we wondered what effect of Heme Oxygenase-1 made on regulation of CD4+CD25+CD127low/- regulatory T cells mediated by mesenchymal stem cells. Peripheral blood mononuclear cells isolated from asthmatic patients and healthy controls were co-cultured with human bone marrow mesenchymal stem cells which were pretreated with Hemin (the revulsive of Heme Oxygenase-1), Protoporphyrin Ⅸ zinc (the inhibitor of Heme Oxygenase-1) and saline. The expression of Heme Oxygenase-1 in MSCs was enhanced by Hemin and inhibited by Protoporphyrin  zinc in vitro. Overexpression of Heme Oxygenase-1 elevated the proportion of CD4+CD25+CD127low/- regulatory T cells in CD4+ T cells, meanwhile, inhibition of Heme Oxygenase-1 decreased the proportion of CD4+CD25+CD127low/- regulatory T cells in CD4+ T cells as compared with mesenchymal stem cells alone. Taken together, these data demonstrated that Heme Oxygenase-1 contributed to the up-regulation of CD4+CD25+CD127low/- regulatory T cells mediated by mesenchymal stem cells in asthma. 

[An in vitro study on the pharmacological ascorbate treatment of influenza virus].

OBJECTIVE: To investigate the effects and mechanism of pharmacological ascorbate against Influenza A/CA/7/09 (H1N12009). METHODS: NHBE cells (≈ 95% confluent monolayer) in 12-well plates (Corning) were kept at 37°C at all times. NHBE cells were exposed to A/CA/7/09 (H1N12009) influenza virus at MOI of 0.01 for 1 h, rinsed with NHBE medium, and incubated with NHBE medium containing 20 mmol/L ascorbate or 20 mmol/L ascorbate +600 IU/ml Catalase. The cells were then incubated for an additional 4 - 12 h and the culture medium was harvested for titration. Viral titers were determined as log(10) 50% tissue culture infective doses (TCID50) assay in MDCK cells. Ascorbate in NHBE medium was determined using HPLC separation coupled with coulometric electrochemical detection. Hydrogen peroxide was detected indirectly by Clark-type oxygen electrode. RESULTS: In vitro experiments showed that pharmacological ascorbate killed not only isolated viruses, but also viruses from normal human bronchial epithelial cells. The antiviral effect of ascorbic acid appeared to be dose-dependent. 2.5 mmol/L ascorbic acid was able to eliminate 90% of the viruses and 20 mmol/L ascorbic acid totally blocked viral replication in vitro. The antiviral effect of pharmacological ascorbate varied at different phases of infection. Pharmacological ascorbate eliminated viral infectivity with treatment times as short as 4 hours at early stage of infection. But the effect was reversed by catalase. CONCLUSION: Pharmacological ascorbate (vitamin C) as a pro-drug eliminates or kills influenza virus, probable by producing steady-state concentrations of hydrogen peroxide (H2O2) in extracellular fluid.

[Effects of shadow boxing training on exercise endurance and quality of life of patients with chronic obstructive pulmonary disease].

OBJECTIVE: To evaluate the effects of shadow boxing training on the exercise endurance and quality of life of Chinese patients with COPD (chronic obstructive pulmonary disease). METHODS: From May 2010 to March 2011, a total of 70 COPD patients in stable phases were recruited from Liwan, Yuexiu and Haizhu districts of Guangzhou. There were 35 patients in the shadow boxing exercise group and 35 patients in the control group. And they were matched by gender and age. The patients in the shadow boxing group exercised for 3 months while those in the control group received the conventional out-hospital management only. Their demographic, medical history, smoking status, medicinal use, spirometric data, clinical COPD questionnaire (CCQ) scores, 6-minute walking distance and Borg scores were collected before and after trial. RESULTS: A total of 63 COPD patients (33 in shadow boxing group vs. 30 in control group) completed the study. There was an average dropout rate of 5.7% (2/35) in shadow boxing group and 14.3% (5/35) in control group. No differences existed between two groups in age (67 ± 8 vs 69 ± 9 yr), male proportion (84.8% vs 86.7%), body mass index (22.8 ± 2.6 vs 22.7 ± 3.0), usage proportion of medicine (42.4% vs 33.3%), duration of disease (4.0 ± 7.5 vs 5.5 ± 7.3), percentage of smokers (78.8% vs 80.0%), 6-minute walking distance (447 ± 94 vs 414 ± 100), CCQ total score (15.0 ± 9.4 vs 14.1 ± 8.8), CCQ symptom score (9.2 ± 5.6 vs 8.3 ± 5.0) and activity score (5.8 ± 4.5 vs 5.8 ± 4.4) at baseline (all P > 0.05). At the end of study, the 6-minute walking distance of patients had statistical differences between two groups (P < 0.01). The shadow boxing group increased by (51 ± 55) m while the control dropped by (19 ± 58) m. The total score, symptom score and activity score of clinical COPD questionnaire had statistical differences between two groups. They decreased significantly in the shadow boxing group as compared with the baseline data while there was no significant change in the control group. No statistical differences existed between two groups in the changes of forced vital capacity (FVC), forced expiratory volume in one second (FEV(1)), FEV(1)% pred, Borg score and dyspnea scales. CONCLUSION: Capable of improving the exercise endurance and life quality of COPD patients, shadow boxing exercise may become one of effective rehabilitation programs for COPD patients in stable phases in communities.

[The effects of nicotine on human airway epithelial cells].

OBJECTIVE: To investigate the genome changes of primary human airway epithelial cells exposed to nicotine in vitro, and therefore to understand the effect of nicotine on the cellular physiological process and phenotypes. METHODS: The primary human airway epithelial cells were divided into 4 groups: 4 h experimental group and control group, 48 h experimental group and control group, with 1×10(8)/L cells in each culture. Total RNA was extracted from cells after incubated with nicotine (1×10(-5) mol/L) for 4 h or 48 h respectively. The genes expressed differentially were detected by a gene chip, and those related to epithelial mesenchymal transition were selected to undergo real-time PCR for verification. RESULTS: Sixty-three up-regulated genes and 44 down-regulated genes were detected in the experimental group incubated with nicotine for 4 h, which were mainly involved in the stress response. There were 860 up-regulated genes and 582 down-regulated genes found in the cells treated with 1×10(-5) mol/L nicotine for 48 h, compared with the control. These genes were mainly involved in some important physiological processes and pathways with transdifferentiation, including embryonic development, cell polarity maintaining, cell adhesion, etc. Further analysis revealed that some epithelial markers such as epithelial keratin and epithelial mucin protein were down-regulated, while mesenchymal cell markers including fiber connecting protein 1 and N-cadherin were up-regulated. The results by real-time PCR showed consistency with those by gene chip examination. CONCLUSION: Nicotine could promote a series of changes in genes related to epithelial mesenchymal transdifferentiation process in human airway epithelial cells.

Asymptomatic patients of chronic obstructive pulmonary disease in China.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) has a variable natural history and not all individuals follow the same course. This study aimed to identify the prevalence and characteristics of asymptomatic COPD patients from a population-based survey in China. METHODS: A multistage cluster sampling strategy was used in a population from seven different provinces/cities. All residents (over 40 years old) were interviewed with a standardized questionnaire and spirometry. Post-bronchodilator forced expiratory volume in 1 second (FEV(1))/forced vital capacity (FVC) of less than 70% was defined as the diagnostic criterion of COPD. All COPD patients screened were divided into symptomatic group and asymptomatic group according to the presence or absence of chronic respiratory symptoms. Socio-demographic, personal and exposure variables were collected and analyzed. RESULTS: Among the 1668 patients who were diagnosed with COPD from the 25 627 sampling subjects, 589 (35.3%) were asymptomatic. The age, sex, body mass index (BMI), rural and urban distributions, smoking habit and education levels were similar in the two groups. A total of 64.7% of the asymptomatic patients had no comorbidities. Cardiovascular diseases and lung cancer were more common among symptomatic COPD patients than asymptomatic group. Asymptomatic COPD group were less likely to present with poor ventilation in the kitchen, a family history of respiratory disease and recurrent childhood cough. Asymptomatic COPD patients had significantly higher FEV(1) (73.1% vs. 61.0%), FVC (91.9% vs. 82.0%), and a higher ratio of FEV(1)/FVC (62.9% vs. 58.7%) (all P < 0.001) than symptomatic group. More asymptomatic patients were underdiagnosed (91.9% vs. 54.3%, P < 0.001) than symptomatic patients. CONCLUSIONS: This large population-based survey confirmed a high prevalence of asymptomatic COPD patients in China. More use of spirometry screening test may be important to the early detection of COPD.

[Effect of adenoviral E1A-involved apoptosis of rat alveolar epithelial cells and human lung adenocarcinoma epithelial cell line].

AIM: The relationship between latent adenovirus infection and apoptosis of airway epithelial cell have not been well documented.We want to illustrating the roles of adenovirus E1A protein on the apoptotic alveolar epithelial in response to TNF-alpha. METHODS: The expression vector for expressing adenovirus E1A protein was transfected into CCL149 and A549 cell respectively. Cell stably expressing E1A protein were selected by G418 resistance.All G418-resistant clones were indentified by RT-PCR and immunocytochemistry. The rate of apoptosis were measured by Hoeschest 33 258 and flow cytometry respectively. The apoptotic rate in response to 30 microg/L TNF-alpha was compared between E1A-positive clones and control clones both in A549 and CCL149. RESULTS: The rates of apoptosis were (2.63+/-0.8)%, (25.38+/-0.9)% respectively in E1A-positive CCL149 cell and (0.62+/- 0.3)%, (6.08+/-0.2)% respectively in E1A-negative CCL149 cell. The rates of apoptosis were (2.63+/-0.8)%, (25.38+/-0.9)% respectively in E1A-positive A549 cell and (0.62+/- 0.3)%, (6.08+/-0.2)% respectively in E1A-negative A549 cell. The rate of apoptosis were increased in E1A-positive cells compared with control with or without TNF-alpha stimulation. CONCLUSION: E1A sensitizes cell to TNF-alpha induced apoptosis of A549 cell and CCL149 cell.

[N-acetyl-L-cysteine inhibits adenoviral E1A-involved transactivation of nuclear factor-kappaB in rat alveolar epithelial cells.].

OBJECTIVE: the relationship between latent adenovirus infection and airway inflammation had not been well documented. The aim of this study was to illustrate the roles of adenovirus E1A protein on the transactivation of NF-kappaB, AP-1 in response to inflammatory stimuli and the effect of N-Acetylcysteine (NAC) upon the transactivation of NF-kappaB and AP-1 in cells stably expressing E1A protein. METHODS: rat alveolar epithelial cells stably expressing adenoviral E1A or control plasmid were developed. For isolation of nuclear extracts, 5 x 10(5) cells were plated and grown overnight in 60 mm dishes. Experiments were repeated 3 times. The cell model of stably expressing adenoviral E1A was stimulated by LPS or TNF-alpha and treated with NAC, a precursor for cysteine. The NF-kappaB and AP-1 transcriptional activity were measured by LUC report system. The expression of NF-kappaB and AP-1 were measured by Western blot. Differences between groups were assessed for significance by Student' t test, and multiple comparisons were made by one-way ANOVA. RESULTS: the luciferase activity derived by NF-kappaB element was (9 698 +/- 98) RLU in untreated E1A-positive clones and (101 195 +/- 234), and (170 385 +/- 443) RLU in LPS and TNF-alpha-stimulated cells, which were significantly higher than that of the control group 2 077 +/- 107, 67 846 +/- 332, 95 743 +/- 211 respectively. The luciferase activity derived by AP-1 element was 9 034 +/- 78 RLU in untreated E1A-positive clones and 26 343 +/- 398 and 31 731 +/- 332 RLU in LPS and TNF-alpha-stimulated cells, which were significantly higher than that of the control group 2 845 +/- 93, 10 772 +/- 432, 11 005 +/- 556 respectively. The densitometry of the NF-kappaB expression in E1A-positive clones were 79.3 +/- 4.6 and 80.3 +/- 3.8 respectively without treatment and were 81.8 +/- 3.9 - 89.9 +/- 1.6 and 94.1 +/- 1.9 to 99.8 +/- 1.6 respectively under LPS or TNF-alpha stimulation, which were significantly higher than that of the control group (68.3 +/- 3.8, 69.4 +/- 4.3 respectively) without stimulation and 70.1 +/- 2.8 to 80.8 +/- 3.6, 73.4 +/- 4.9 to 83.2 +/- 6.7 respectively under stimulation. The level of AP-1 expression did not show difference upon treatment with LPS or TNF-alpha in either cell clones. The densitometry of the NF-kappaB expression in E1A-positive clones were 3.2 +/- 0.1 and 3.3 +/- 0.1 respectively under LPS and TNF-alpha-stimulation and 1.98 +/- 0.2 and 1.9 +/- 0.2 respectively upon treatment for LPS and TNF-alpha with NAC pre-incubation. CONCLUSIONS: these results indicate that E1A protein upregulated NF-kappaB transcription activity induced by LPS and TNF-alpha in rat alveolar epithelial cells and this effect could be repressed by NAC. The mechanisms underlying transactivation of NF-kappaB involved by E1A may be related to oxidative stress.

[The oxidative stress enhanced adenoviral E1A-involved transactivation of nuclear factor-kappaB].

OBJECTIVE: The relationship between latent adenovirus infection and airway inflammation have not been well documented. The aim of this study is to illustrate the roles of adenovirus E1A protein on the level of glutathione (GSH) in response to oxidative stress and the effect of the oxidant/antioxidant imbalance upon the transactivation of NF-kappaB triggered by E1A protein. METHODS: Rat alveolar epithelial cell stably expressing adenoviral E1A or control plasmid were developed. For isolation of nuclear extracts, 5 x 10(5) cells were plated and grown overnight in 60 mm dishes. Experiments were repeated three times. The cell model of stably expressing adenoviral E1A was stimulated by H2O2. The level of GSH were measured. E1A positive clone was stimulated by LPS or TNF-alpha and treated with L-Buthionine-sulfoximine (BSO). The expression of NF-kappaB was measured by Western blot. Differences between groups were assessed for significance by Student' t test; multiple comparisons by the one-way ANOVA. RESULTS: There is no difference of GSH level without stimulation between E1A-positive clones and E1A-negative clones. For E1A-positive clones, the level of GSH did not increase in response to H2O2 as E1A-negative clones. The quantitation by densitometry of the NF-kappaB expression in E1A-positive clones were (79.3 +/- 4.6), (80.3 +/- 3.8) respectively without treatment and were (81.8 +/- 3.9) - (89.9 +/- 1.6) and (94.1 +/- 1.9) - (99.8 +/- 1.6) respectively under LPS or TNF-alpha stimulation, which were significantly higher than that of the control group (68.3 +/- 3.8), (69.4 +/- 4.3) respectively without stimulation and (70.1 +/- 2.8) - (80.8 +/- 3.6), (73.4 +/- 4.9) - (83.2 +/- 6.7) respectively under stimulation. The quantitation by densitometry of the NF-kappaB expression in E1A-negative clones were (1.25 +/- 0.18) and (1.69 +/- 0.19) respectively under LPS and TNF-alpha-stimulation and (1.22 +/- 0.16) and (1.75 +/- 0.13) respectively upon treatment for LPS and TNF-alpha with BSO preincubation. There did not show difference upon treatment with LPS or TNF-alpha with or without BSO in E1A-negative cell clone. The quantitation by densitometry of the NF-kappaB expression in E1A-positive clone were (1.75 +/- 0.10) and (2.26 +/- 0.21) respectively upon treatment for LPS and TNF-alpha with BSO preincubation which were significantly higher than that of LPS or TNF-alpha-stimulation alone (1.35 +/- 0.12), (1.80 +/- 0.14) respectively. CONCLUSION: These results indicate that E1A protein decreased GSH levels in oxidant stress and upregulated NF-kappaB transcription activity. The oxidant/antioxidant imbalance in rat alveolar epithelial cells enhances E1A-modulated transcriptional activation of NF-kappaB. The mechanism underlying transactivation of NF-kappaB involved by E1A may be related to oxidative stress.

[Current status of prevention and management of chronic obstructive pulmonary disease in rural area in China].

OBJECTIVE: To investigate the current status of prevalence, prevention and management of chronic obstructive pulmonary disease (COPD) in rural area in China. METHODS: A cross-sectional survey of COPD was conducted in Beijing city, Shanghai city, Guangdong province, Liaoning province, Tianjin city, Chongqing province and Shanxi province. A population-based cluster sample was randomly selected from each rural area. In the selected community, all residents at least 40 years old were recruited, and interviewed with a modified standardized questionnaire from the international burden of obstructive lung diseases (BOLD) study. All participants were tested with spirometry. Those with airflow limitation were performed on post-bronchodilator spirometry. The post-bronchodilator a ratio of forced expiratory volume in one second to forced vital capacity (FEV1/FVC) less than 70% was defined as the diagnostic criteria of COPD. RESULTS: (1) Data of 9434 participants was valid for analysis, with a valid response rate of 83.6%; the prevalence of COPD in rural was 8.8% (830/9434), 12.8% in male and 5.4% in female. (2) The percentage of smoking and the exposure to biomass smoke in rural was 43.0% (4059/9434) and 83.1% (7835/9434) respectively; cigarettes cessation rate was 17.5%; only 12.4% (502/4059) of smokers had received advice to quit smoking. (3) Among COPD patients, only 30.0% (249/830) had ever been diagnosed as COPD, bronchitis, emphysema, or asthma, 2.4% (20/830) had ever received spirometric tests, and 74.5% were current smokers; only 7.9% (50/634) COPD patients in stage two or over had received regular drug treatment. CONCLUSION: There was high prevalence and poor prevention and management for COPD in rural areas. Therefore, an enforced prevention and management for COPD are urgent.

[The effects of adenovirus 5 latent infection on pulmonary inflammation caused by biomass fumes in guinea pigs].

OBJECTIVE: To explore the effects of adenovirus 5 (Ad5) latent infection on oxidized injury and inflammation of the lungs caused by exposure to biomass fumes in guinea pigs. METHODS: Forty-six albino guinea pigs were randomly divided into 2 groups. One group (n = 26) was infected intranasally with Ad5, and the other group (n = 20) was sham-infected with sterile PBS as control. After 22 days, the living animals (n = 20 each) were randomly divided into 2 subgroups. One subgroup was exposed to biomass fumes for 3 weeks, and the other subgroup was exposed to room air. At the end of the experiment, all animals were sacrificed and their lung tissues were examined histopathologically. The levels of interleukin (IL)-8, IL-6, and cell adhesion molecule-1 (CAM-1) in bronchoalveolar lavage fluid (BALF) were measured. The results were analyzed using a two-way analysis of variance (ANOVA) with two crossed factors. The level of significance was P < 0.05. RESULTS: The exposure to biomass fumes and the latent infection of Ad5 were associated with a significant increase in the total number of airway inflammatory cells in the BALF [(37.1 +/- 5.5) x 10(6)/L and (16.8 +/- 2.3) x 10(6)/L], F = 208.947, 22.687, all P < 0.01). The exposure to biomass fumes was associated with a significant increase in the concentration of IL-8, IL-6, and CAM-1 in the BALF [(0.38 +/- 0.06), (0.188 +/- 0.021) and (6.5 +/- 1.6) mg/kg], F = 13.525 - 69.021, all P < 0.01). The latent infection of Ad5 was associated with a significant increase in the concentration of IL-8, and CAM-1 in the BALF [(0.37 +/- 0.05) and (8.2 +/- 2.1) mg/kg] (F = 11.964, 57.162, all P < 0.01). Ad5 infection had a synergistic effect on the IL-8 and CAM-1 production caused by biomass fume exposure. CONCLUSIONS: Biomass fumes caused severe histopathological changes and inflammation in lungs of guinea pigs, and Ad5 latent infection aggravated these changes. Increased production of inflammatory cytokines may play an important role in this process.

[The mechanism of inhibition by ginkgolide B on interleukin-8 production induced by 4-hydroxynonenal in bronchial epithelial cells].

OBJECTIVE: 4-Hydroxynonenal (4-HNE) can increase the synthesis of interleukin-8 (IL-8) in bronchial epithelium cells (16HBE). This study was to explore the role of ginkgolide B in inhibiting the synthesis of IL-8 induced by 4-HNE in 16HBE. METHODS: The experiments were divided into 3 groups: a group treated with 4-HNE (10 micromol/L), a group treated with ginkgolide B (100 micromol/L) + 4-HNE (10 micromol/L), and a control group. IL-8 and IL-8 mRNA were measured after 4-HNE (or serum-free medium) stimulation for 0.5, 2, 4, 8, 12 hours. The phosphorylation of ERK1/2, JNK, p38MAPK and the combining activity of AP-1 after 4-HNE stimulation for 0.5, 2, 4, 8, 12 hours were all examined. IL-8 and the combining activity of AP-1 were measured after the 16HBE were pre-incubated with 50 micromol/L PD98059 (MEK1 inhibitor) for 2 hours before 4-HNE stimulation. The combining activity of AP-1 in 10 micromol/L 4-HNE, 100 micromol/L ginkgolide B + 10 micromol/L 4-HNE, and the control groups were all measured by EMSA. RESULTS: The level of IL-8 in 10 micromol/L 4-HNE, 100 micromol/L ginkgolide B + 10 micromol/L 4-HNE, the control groups after 4-HNE stimulating for 4 h were (98.3 +/- 4.2), (88.2 +/- 5.3), (65.3 +/- 6. 2) and (116.5 +/- 5.6), (102.8 +/- 4.7), (63.7 +/- 6.6) microg/L for 12 h. The level of IL-8 and IL-8 mRNA after 4-HNE stimulation in the ginkgolide B + 4-HNE group were lower than those in the 4-HNE group while higher than those in the control groups. The level of phosphorylation of ERK1 in the 4-HNE group at 0.5, 2, 4, 8, 12 hours were higher than those in the control groups (t = 2.83 - 14.03, P < 0.05). The AP-1 combining activity in the 4-HNE group, the ginkgolide B + 4-HNE group, PD98059 + 4-HNE group, and the control group were significantly different (F = 21.49 - 194.16, P < 0.01). The expression of IL-8 and the AP-1 combining activity in groups of pre-incubated with PD98059 2 hours before 4-HNE stimulation were lower than that without PD98059. The combining activity of AP-1 in the ginkgolide B + 4-HNE group was decreased as compared to the 4-HNE groups. CONCLUSION: 4-HNE increased the expression of Interleukin-8 in bronchial epithelium cells, via increasing the transcription activities of AP-1 by ERK1 cell signal transduction pathways. Ginkgolide B inhibited synthesis of IL-8 by blocking ERK1-AP1 transduction pathways.

[The effect of adenoviral E1A on inflammatory mediator expression of rat alveolar epithelial cells].

OBJECTIVE: The relationship between latent adenvorius infection and airway inflammation has not been well documented. The aim of this study is to illustrate the roles of adenovirus E1A protein on the inflammation mediator expression in response to lipopolysaccharide and tumor necrosis factor alpha (TNF-alpha) in rat alveolar epithelial cells. METHODS: An eukaryotic expression vector for expressing adenovirus E1A protein was constructed and transfected into CCL149 cells. Cells stably expressing E1A protein were selected by G418 resistance. The inflammatory mediator intercellular adhesion molecule-1 (ICAM-1) expression in response to lipopolysaccharide and TNF-alpha was compared between adenovirus E1A-positive clones, control clones and CCL149 cells. Messenger RNA of ICAM-1 was measured by RT-PCR, and proteins quantified by flow cytometry. The nuclear factor kappaB (NF-kappaB) and activator protein 1 (AP-1) activity were measured by LUC report system and electrophoretic mobility shift assay (EMSA). RESULTS: ICAM-1 messenger RNA and protein were increased in E1A-positive cells exposed to 10 ng/ml TNF-alpha and 10 microg/ml lipopolysaccharide. The luciferase activity drived by NF-kappaB and AP-1 elements were increased in E1A-positive cells compared with control with or without lipopolysaccharide and TNF-alpha stimulation. EMSA showed that only NF-kappaB activity increased in E1A-positive cells. This increase was not observed in AP1 element drived EMSA. CONCLUSIONS: The results indicate that E1A upregulates ICAM-1 expression induced by lipopolysaccharide and TNF-alpha in rat alveolar epithelial cells. E1A enhances the expression of inflammatory mediator by triggering NF-kappaB activity. It is suggested that E1A amplified inflammatory response may contribute to the pathogenesis of chronic obstructive pulmonary disease.

[Association of tumor necrosis factor alpha 308 G/A gene promoter polymorphism with the presence of chronic obstructive pulmonary disease: a meta-analysis].

OBJECTIVE: To investigate the association between the polymorphism of tumor necrosis factor alpha 308 gene (TNF-alpha-308) promoter and the risk of chronic obstructive pulmonary disease (COPD) using the method of meta-analysis. METHOD: The database of Medline and the Chinese biomedicine disc (CBM) were searched for published case-control studies of the association between the polymorphism of TNF-alpha-308 gene promoter and COPD. Data were extracted using a standardized form and the meta-analysis was performed. RESULTS: Eighteen case-control studies, comprising 1606 patients with COPD and 2551 controls (the oriental population: 666 patients with COPD and 898 controls; the Caucasian: 940 patients with COPD and 1653 controls) were included in the meta-analysis. Using a fixed effect model, the pooled result in the oriental population showed that the TNF2 allele was associated with the susceptibility to COPD [odds ratio (OR) = 2.62, 95% confidence interval (95% CI) 2.00 to 3.43]. The OR for COPD susceptibility in TNF1/2 population was significantly increased at 2.44 (95% CI 1.79 to 3.33) compared to the TNF1/1 population. The OR for COPD susceptibility in the TNF1/2 plus TNF2/2 population was significantly increased at 2.78 (95% CI 2.06 to 3.75) compared to the TNF1/1 population. When adjusted for smoking, the result was similar. However, in the Caucasian population, the TNF2 allele was not associated with the susceptibility to COPD (OR = 0.97, 95% CI 0.84 to 1.14). There was no association between the genotype of TNF-alpha-308 and COPD (OR = 0.96, 95% CI 0.79 to 1.16, and OR = 1.03, 95% CI 0.86 to 1.25), compared between the TNF1/2 population, the TNF1/2 plus the TNF2/2 population and the TNF1/1 population respectively. When adjusted for smoking, the result was similar. CONCLUSION: In the oriental population, the TNF2 allele confers a significant risk for developing COPD. There is no association between the polymorphism of TNF-alpha-308 gene promoter and COPD in the Caucasian population.

[A study on the correlation of body mass index with chronic obstructive pulmonary disease and quality of life].

OBJECTIVE: To investigate the correlation between body mass index (BMI) and chronic obstructive pulmonary disease (COPD), based on a cross-sectional COPD survey conducted in Beijing, Shanghai, Guangdong, Liaoning, Tianjin, Chongqing and Shaanxi of China between 2002-2004. METHODS: A multi-stage stratification cluster sampling strategy was used in this cross-sectional survey, and 20,245 subjects (8705 males and 11,540 females) aged 40 years or older were recruited, interviewed with a questionnaire, measured for height and weight, and tested with spirometry. 1668 subjects with post-bronchodilator FEV1/FVC less than 70% were identified as having COPD after other known causes of airflow limitation were excluded. Analysis on relationship between COPD and BMI was performed in 1668 COPD and 18 577 non-COPD subjects. RESULTS: Compared with non-COPD subjects, BMI was significantly lower in COPD patients [(22.7+/-3.5) vs (24.1+/-3.4) kg/m2, F=158.31, P<0.01]; BMI was also significantly lower in smokers than in non-smokers [(23.6+/-3.4) vs (24.2+/-3.5) kg/m2, F=49.10, P<0.01]. And an addictive interaction to BMI between COPD and smoking was observed (F=6.03, P<0.05). BMI decreased with the increase of the stage of COPD (F=45.6, P<0.01), with a negative relationship (r=-0.08, P<0.01). Lower BMI was significantly associated with increased risk of COPD (chi2=102.68, P<0.01). Compared with subjects with normal BMI (BMI=24.0-27.9 kg/m2), those with lower BMI (BMI<18.5 kg/m2) were more likely to have COPD [adjusted OR=2.12 (95% CI 1.73-2.59)], while those with higher BMI (BMI=24.0-27.9 kg/m2) and obesity (BMI>or=28.0 kg/m2) were less likely to have COPD [adjusted OR=0.67 (95% CI 0.59-0.76); and 0.60 [(95% CI 0.49-0.73), respectively]. Moreover, there was an interaction to COPD between smoking and BMI (chi2=4.73, P<0.05). Compared with COPD patients with normal BMI, the quality of life in those with lower BMI was impaired (55+/-8 vs 57+/-6 in mental scores of SF-12, F=2.96, P<0.05; 42+/-10 vs 46+/-9 in physical scores of SF-12, F=4.21, P<0.01), and their dyspnea scores were higher (1.4+/-1.5 vs 1.1+/-1.3, chi2=14.32, P<0.01). CONCLUSION: Lower BMI was strongly associated with COPD, possibly as a risk factor for COPD independent of smoking, and a potential predictor for COPD severity.

[Effect of allitridum on expression of the gamma-glutamyl-cysteine synthetase in rat lung epithelial L2 cells].

OBJECTIVE: To study the effects of allitridum on the antioxidative capability of rat lung epithelial L2 cells (CCL-149 cell line) by observing the changes of glutathione (GSH), malondialdehyde (MDA) content, and the expressing level of gamma-glutamyl-cysteine synthetase (gamma-GCS) protein. METHODS: The cell viability of CCL-149 cells treated with allitridum was detected by MTT assay. CCL-149 cells were incubated with 0, 0.1, 1.0, 5.0 microg/ml of allitridum for 6, 12, 24, 48 h. After treatment, GSH content and MDA formation were measured by spectrophotometric assay, and the gamma-GCS protein was semi-quantified by Western blot. RESULTS: Allitridum (0.1, 1.0, 5.0 microg/ml) showed no side effects on cell growth (P > 0.05). Treatment with allitridum increased GSH levels and decreased MDA formation in CCL-149, the most significant time being incubation at 24 h. The GSH content were (13.34 +/- 0.62), (27.67 +/- 2.39), (29.54 +/- 0.71), (30.25 +/- 1.05) mg/g prot, and the MDA content were (1.25 +/- 0.08), (0.90 +/- 0.06), (0.84 +/- 0.06), (0.81 +/- 0.02) nmol/mg prot when CCL-149 cells were treatment with 0, 0.1, 1.0, 5.0 microg/ml of allitridum for 24 h, respectively. The effect showed no dose dependent effects (P > 0.05). After 6, 12, 24 h incubation, the expression of gamma-GCS protein was increased as compared to the control cells (t = 2.82 - 9.92, P < 0.05). CONCLUSION: Allitidum may increase the antioxidative ability of CCL-149 cells by increasing the expression of gamma-GCS protein and the content of GSH.