Showing papers in "Chinese Journal of Tuberculosis and Respiratory Diseases in 2005"

Early use of noninvasive positive pressure ventilation for patients with acute exacerbations of chronic obstructive pulmonary disease:a multicentre randomized controlled trial

Abstract: Objective To assess the efficacy and safety of noninvasive positive pressure ventilation(NPPV) for acute exac erbations of chronic obstructive pulmonary disease(AECOPD)patients on general wards. Methods A prospective multicentre randomized contro lled trial was conducted in 19 hospitals in China over 16 months. 42 AECOPD pa tients with pH≥7.25 and arterial partial pressure of carbon dioxide(PaCO2) 45 mm Hg (1 mm Hg=0.1 kPa)were recruited on general wards and randomly assign ed to standard medical treatment(group A,n=171) or early administration o f NPPV plus standard medical treatment(group B,n=171). Results The characteristics of the two groups on admission were similar. Th e number of AECOPD patients requiring intubations in group B was significantly l ess than that of the group A(8/171,26/71,P=0.002). Subgroup analysis sh owed the need for intubation in both the mild(pH≥7.5) and the severe(pH7 .0) acidotic patients in group B were decreased(9/80,2/71,P=0.047 and 8/0,/4,P=0.048 respectively). The mortality in hospital was reduced s lightly by NPPV but with no significant difference(7/171,12/171,P=0.45) . Respiratory rate(RR),scale for accessory muscle use and arterial pH improv ed rapidly at the first 2 hours only in patients of group B. After 24 hours,th e differences of pH,arterial partial pressure of oxygen(PaO2),scale for access ory muscle use and RR in group B[7.6±0.06,(72±22)mm Hg,2.5±0.9,(2 2±4)/min]were statistically significant compared with group A[7.7±0.05, (85±4)mm Hg,2.±1.1,(21±4)/min,P0.01 for all comparisons]. Conclusions The early use of NPPV on general wards improve s arterial blood gas and respiratory pattern,decreases the rate of need for int ubation in AECOPD patients. NPPV is indicative for alleviating respiratory musc le fatigue and preventing respiratory failure from exacerbation.

Simvastatin induces eosinophil apoptosis in vitro

Abstract: Objective To investigate the effect of simvastatin, a 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibitor, on eosinophils (EOSs) apoptosis in asthma patients. Methods Peripheral blood EOSs from 10 asthma patients were cultured in the presence or absence of simvastatin (1, 5, 10, 20 micromol/L), together with or without mevalonate (100 micromol/L) for 6, 12, 24, and 48 h. Apoptosis was monitored by annexin V/PI staining and flow cytometry. Caspase-3 was measured by enzyme-linked immunosorbent assay (ELISA). Results EOSs were particularly susceptible to apoptosis after incubated with 5 micromol/L simvastatin for 6, 12, 24, and 48 h [the rates of EOSs undergoing apoptosis were: (23 +/- 3)%, (24 +/- 3)%, (41 +/- 6)%, (70 +/- 12)% in control and (32 +/- 4)%, (47 +/- 7)%, (62 +/- 9)%, (86 +/- 14)% in simvastatin; compared with control at the same time point: P = 0.000]. EOS apoptosis occurred at doses of 1 micromol/L and was already maximal at 5 micromol/L after incubated with simvastatin for 12 h [the rates of EOSs undergoing apoptosis were: (24 +/- 3)% in control, (37 +/- 3)%, (51 +/- 3)%, (53 +/- 4)%, (52 +/- 4)% in 1, 5, 10, 20 micromol/L simvastatin, respectively; compared with control: P = 0.000]. The level of caspase-3 in EOSs was consistent with the rate of cell apoptosis [(8 +/- 3) microg/L in control, (14 +/- 4), (22 +/- 4), (24 +/- 4), (23 +/- 5) microg/L in 1, 5, 10, 20 micromol/L simvastatin, respectively; compared with control: P = 0.000 - 0.003]. However, Co-incubation of simvastatin with mevalonate (the production of HMGR) completely reversed the activity of simvastatin on EOS apoptosis even when the highest simvastatin (20 micromol/L) dose was used; the rates of EOSs undergoing apoptosis in the control, mevalonate plus simvastatin and simvastatin alone were (24 +/- 3)%, (52 +/- 4)% and (25 +/- 3)%, respectively; while the caspase-3 levels were (8 +/- 3) microg/L, (23 +/- 5) microg/L and (9 +/- 3) microg/L, respectively. Conclusion Simvastatin induces apoptosis of EOSs in asthma patients via its ability to block the synthesis of the important isoprenoid intermediates, which leads to the inhibition of small GTP-binding protein activity.

Evaluation of the mycobacterial interspersed repetitive units typing as a practical approach in molecular epidemiology of Mycobacterium tuberculosis

Abstract: OBJECTIVE To introduce a new genotyping method, mycobacterial interspersed repetitive units (MIRU) typing, for Mycobacterium tuberculosis and to evaluate its feasibility. METHODS Mycobacterium tuberculosis strains stored at Shanghai Municipal Center for Disease Control and Prevention during 2000 to 2002, were randomly selected by simple digital table and genotyped by MIRU and Spoligotyping. RESULTS By Spoligotyping method, 91 strains were typed to 20 genotypes, of which 89% (81/91) strains belonged to Beijing genotype, while by MIRU method, these strains were divided into 46 genotypes. The MIRU typing showed high discriminatory power, especially for the Beijing genotype strains. The 81 Beijing genotype strains could be subdivided into 39 different MIRU genotypes. In this sample collection, 12 MIRU loci showed different discriminative according to their allelic diversity. Locus 26 showed highly discriminative, while locus 16, 31, and 40 showed moderately discriminative. CONCLUSIONS MIRU genotyping is a simple and fast method. Its numerical result facilitates the comparison among strains of Mycobacterium tuberculosis from different labs.

[The prevalence of obstructive sleep apnea-hypopnea syndrome in adults aged over 20 years in Changchun city].

Abstract: OBJECTIVE: To investigate the prevalence of obstructive sleep apnea-hypopnea syndrome (OSAHS) in adults aged over 20 years in Changchun city, providing epidemiological data for treatment and prevention of the disease. METHODS: 3,960 subjects were derived from a stratified cluster and random sampling of the population in two districts of Changchun city. They were asked to answer the questions from a questionnaire in their houses. According to the degree of snoring, 200 subjects with a snoring score >or= 2 degree were selected to undergo polysomnography for a whole night and the prevalence of the disease was estimated. RESULTS: A total of 3,648 (97.64%) validated questionnaires was collected. Of them 31.00% had a snoring score >or= 2 degree, the prevalence was higher in males (40.07%) than in females (21.76%). The prevalence of snoring was higher in drivers (42.47%) than in other occupations. The estimated prevalence of OSAHS defined by apnea-hypopnea index (AHI) >or= 5, Epworth sleepiness scale (ESS) >or= 9 and arterial oxygen saturation (SaO(2)) < 90% was 4.81%. CONCLUSIONS: The estimated prevalence of OSAHS in adults aged over 20 years in Changchun city was 4.81%. Doctors should pay more attention to the disease and the ordinary people should be informed of the health impact of snoring and OSAHS.

Gene transfection of Livin isoforms into A549 cell line and its effect on cell growth and sensitivity to chemotherapy and radiotherapy

Abstract: OBJECTIVE To express Livin alpha & beta in A549 cells by using gene transfection, and to observe its effect on cell growth and cell sensitivity to chemotherapy drugs and radiation METHODS Eukaryotic expression vectors of Livin alpha & beta were transfected into A549 cells and cell clones with stable expression were obtained Livin alpha & beta expression levels in the transfected A549 cells were assessed at mRNA level and protein level, respectively Cell growth status was assessed by biological features MTT was performed to test effects of Livin on sensitivity of the A549 cells to chemotherapy drugs and radiation, and cell cycle analysis was performed to evaluate cell apoptosis RESULTS After transfection, positive cells, especially A549 cells expressing Livin, showed an increase of about 20% in colony-forming ability, a shorter doubling time (P < 005) and lower sensitivity to chemotherapy drugs and radiation (P < 001) Only 02% of the cells committed apoptosis with 10 Gy radiation CONCLUSION Livin isoforms, especially Livin alpha, are implicated in genesis and development of lung cancer, thus may be an important mechanism for drug resistance of lung cancer cells

Characteristics of airway inflammatory cells and mediators in eosinophilic bronchitis patients

Abstract: OBJECTIVE To investigate the features of airway inflammation in patients with eosinophilic bronchitis (EB) by analyzing the inflammatory cells and mediators in induced sputum and bronchoalveolar lavage fluid (BALF). METHODS Sputum induced by hypertonic saline aerosol inhalation was collected in 43 patients with EB (EB group), 20 patients with cough variant asthma (CVA, CVA group), 16 patients with bronchial asthma (asthma group) and 21 healthy controls (healthy group). Bronchoalveolar lavage was also performed in 11 patients with EB and 10 patients with CVA. Differential cell count was carried out in sputum and BALF. Levels of eosinophilic cationic protein (ECP), leukotriene C(4) (LTC(4)) and histamine in sputum and BALF were measured. RESULTS The percentage of sputum eosinophils (EOS) showed significant difference among the four groups; healthy group 0.0020 +/- 0.0050, EB group 0.1130 +/- 0.1470, CVA group 0.1900 +/- 0.1800, asthma group 0.3860 +/- 0.2670 (P < 0.01). The difference between asthma group and CVA group, and the difference between CVA group and EB group were significant (P < 0.05). The percentage of EOS in BALF was (0.011 +/- 0.016) in EB group, (0.053 +/- 0.040) in CVA group, the difference being significant (P < 0.05). The concentration of sputum ECP was (0.62 +/- 0.66) mg/L in EB group, (1.27 +/- 1.74) mg/L in CVA group, (0.07 +/- 0.10) mg/L in healthy group, the difference among the three groups being significant (P < 0.01). The difference of LTC(4) level was also significant when CVA group (0.65 +/- 0.62) microg/L was compared with EB group (0.39 +/- 0.61) microg/L (P < 0.05) and healthy group (0.15 +/- 0.11) microg/L (P < 0.01). The difference of histamine level in the supernatant of BALF was significant between CVA group (3.4 +/- 1.4) microg/L and EB group (1.6 +/- 1.5) microg/L (P < 0.05). CONCLUSIONS EOS infiltration is mainly localized to the central airway in EB, with lower airway levels of LTC(4) and histamine as compared to CVA. These inflammatory features may partly explain the absence of non-specific airway hyperresponsiveness in patients with EB.

[Characterization of the katG, inhA, ahpC, kasA, and oxyR gene mutations in isoniazid-resistant and susceptible strain of Mycobacterium tuberculosis by automated DNA sequencing].

Abstract: OBJECTIVE To study the characteristics of katG, inhA, ahpC, kasA, and oxyR gene mutations in isoniazid-resistant clinical isolates of Mycobacterium tuberculosis METHODS A total of 101 isoniazid-resistant and 43 susceptible strains of Mycobacterium tuberculosis were analyzed by PCR and sequence analysis of their katG, inhA, ahpC, kasA, and oxyR genes RESULTS (1) Sequencing of katG from 101 INH-resistant strains showed point mutations, small deletions or insertions in 81 isolates (802%), but no complete deletions were identified The mutations at 16 position were found for the first time Point mutations at position 315 were found in the genomes of 386% (39/101) of isoniazid-resistant strains Low level isoniazid resistant strains (1 microg/ml) had higher mutation frequency at 315-Ser than high level isoniazid resistant strains (10 microg/ml; chi(2) = 931, P < 005) Mutations at position 463 were detected in 58 (574%) isoniazid-resistant strains Arg463leu was also present in 23 of 43 susceptible strains (2) Mutations in inhA genes were identified in 5 isoniazid-resistant isolates (49%) None of the susceptible strains contained any mutation in inhA genes (3) Only 3 isolates in the 101 (297%) isoniazid-resistant clinical isolates had mutations in ahpC genes No mutations were identified in the ahpC genes in 43 isoniazid-susceptible isolates (4) Mutations in kasA genes were present in 17 of 101 (168%) isoniazid-resistant isolates However, G312S was also present in 3 of 43 susceptible strains (5) None of the isoniazid-resistant strains and susceptible isolates contained oxyR gene mutation (6) Taken together, 91 of 101 (90%) isoniazid-resistant strains had mutations in katG, inhA, ahpC, and kasA genes which were associated with drug resistance CONCLUSION These studies provide further evidence supporting the association between katG, inhA, ahpC, and kasA gene mutations and INH resistance in Mycobacterium tuberculosis, while other mechanisms of INH resistant may exist

[Effect of a new gasotransmitter, hydrogen sulfide, on collagen remodeling of pulmonary artery under hypoxia].

Abstract: OBJECTIVE To study the modulatory effect of hydrogen sulfide (H(2)S) on the accumulation of collagen type I and type III in the wall of pulmonary small artery during hypoxic pulmonary vascular remodeling. METHODS Nineteen male Wistar rats were randomly divided into a control group (n = 6), a hypoxic group (n = 7) and a hypoxia + NaHS group (n = 6). Hypoxic challenge was performed everyday for 21 days. NaHS solution was injected peritoneally everyday before hypoxia challenge for rats in the hypoxia + NaHS group. After 21 days of hypoxia, the mean pulmonary artery pressure was measured by pulmonary artery catheterization. The weight ratio of right ventricle to left ventricle + septum [RV/(LV + SP)] was also measured. The plasma level of H(2)S was determined by methylene blue spectrophotometric method. The expression of collagen type I and type III in pulmonary small arteries were detected by immunohistochemistry. The expression of procollagen type I and type III mRNA in pulmonary small arteries were detected by in situ hybridization. RESULTS (1) Compared with the control group, the mPAP increased by 46% (P < 0.01), the weight ratio of RV/(LV + SP) increased by 41% and the plasma level of H(2)S decreased by 36% for rats in the hypoxia group (P < 0.01). Compared with the hypoxia group, the mPAP decreased by 31% (P < 0.01), the weight ratio of RV/(LV + SP) decreased by 24% and the plasma level of H(2)S increased by 65% (P < 0.01) for rats in the hypoxia + NaHS group. (2) Expression of collagen type I in small and median pulmonary arteries of the three groups: compared with rats in the control group, collagen type I expression increased by 81% and 62% respectively for rats in the hypoxia group (P < 0.01); compared with rats in the hypoxia group, the expression decreased by 32% and 18% respectively for rats in the hypoxia + NaHS group (P < 0.01). (3) Expression of procollagen type I mRNA in small and mid pulmonary arteries of the three groups: compared with rats in the control group, the expression increased by 49% and 68% respectively (P < 0.01) for rats in the hypoxia group; compared with rats in the hypoxia group, the expression decreased by 31% and 33% respectively for rats in the hypoxia + NaHS group (P < 0.01). (4) Expression of collagen type III in small pulmonary arteries of the three groups: compared with rats in the control group, the expression increased by 84% for rats in the hypoxia group (P < 0.01); compared with rats in the hypoxia group, the expression decreased by 37% for rats in the hypoxia + NaHS group (P < 0.01). Compared with rats in the control group, the expression of collagen type III in median pulmonary arteries increased by 38% in the hypoxia group (P < 0.01), while there was no significant difference between the hypoxia group and the hypoxia + NaHS group. (5) Expression of procollagen type III mRNA in small and median pulmonary arteries of the three groups: compared with rats in the control group, the expression increased by 53% and 17% respectively (P < 0.01) for rats in the hypoxia group; compared with rats in the hypoxia group, the expression decreased by 45% and 33% respectively for rats in the hypoxia + NaHS group (P < 0.01). CONCLUSIONS In the process of hypoxic pulmonary vascular collagen remodeling in rats, H(2)S could inhibit the abnormal accumulation of collagen type I and type III in the wall of pulmonary small arteries. This effect may be one of the mechanisms by which H(2)S ameliorates hypoxic pulmonary vascular remodeling.

[Effects of Shen-Mai injection on sternohyoid contractile properties in chronic intermittent hypoxia rat].

Abstract: OBJECTIVE To investigate the effect of Shen-Mai injection (SMI) on sternohyoid contractile properties in rats with chronic intermittent hypoxia METHODS Thirty healthy male SD rats were randomly assigned to three equal groups, the control group (A group), the chronic intermittent hypoxia group (B group) and the SMI group(C group) Rats in B group and C group were exposed to alternating periods of hypoxia and normoxia once per minute for 8 h/d for 5 weeks in order to mimic the intermittent hypoxia of obstructive sleep apnea-hypopnea syndrome (OSAHS) in humans Isometric contractile properties were determined by electrostimulating the strips of isolated sternohyoid muscles at different frequencies (from 10 Hz to 100 Hz) to observe the changes of the sternohyoid contractile properties RESULTS (1) The tension of sternohyoid muscle in A group at different frequencies was (232 +/- 56), (262 +/- 50), (351 +/- 54), (460 +/- 85), (570 +/- 99), (699 +/- 97), (792 +/- 95), (857 +/- 76), (879 +/- 79), and (866 +/- 124) g/cm(2) The tension of sternohyoid muscle in B group [(195 +/- 47), (238 +/- 47), (330 +/- 51), (451 +/- 59), (542 +/- 70), (661 +/- 91), (742 +/- 91), (797 +/- 90), (820 +/- 84), and (807 +/- 118) g/cm(2)] was not significantly different from those in A group respectively (all P > 005); while the tension of sternohyoid muscle in C group [(305 +/- 23), (400 +/- 54), (562 +/- 76), (722 +/- 64), (820 +/- 55), (924 +/- 46), (981 +/- 40), (992 +/- 74), (1018 +/- 39), and (1022 +/- 40) g/cm(2)] was significantly different from those in B group respectively (all P < 005) (2) In fatigue test, the tension percentages of sternohyoid muscle in A group at 1, 2, 3, 4, and 5 min were (879 +/- 57)%, (721 +/- 115)%, (556 +/- 96)%, (397 +/- 107)%, (332 +/- 102)% Compared with A group, the tension percentages of sternohyoid muscle in B group at 1, 2, 3, 4, and 5 min [(756 +/- 85)%, (416 +/- 73)%, (290 +/- 27)%, (204 +/- 29)%, (185 +/- 25)%, respectively] decreased significantly (all P < 005) Compared with B group, the tension percentages of sternohyoid muscle in C group [(879 +/- 44)%, (679 +/- 141)%, (484 +/- 99)%, (382 +/- 70)%, (338 +/- 93)%, respectively] increased significantly (all P < 005) CONCLUSIONS Chronic intermittent hypoxia can increase upper airway muscle fatigue SMI can significantly increase the contractile properties of upper airway muscle and resist the fatigue of upper airway muscle

Serum resistin and leptin in patients with chronic obstructive pulmonary disease and their relationship to nutritional state

Abstract: OBJECTIVE To investigate the serum resistin and leptin levels, their relationship to nutritional state and the associated factors in patients with chronic obstructive pulmonary disease (COPD). METHODS The serum resistin and leptin levels in 57 stable COPD patients and 31 healthy controls were measured by enzyme-linked immunosorbent assay (ELISA) and radio-immunoassay respectively. Correlated factors of serum resistin and leptin were analyzed. RESULTS The serum resistin and leptin levels in COPD patients [(2.1 +/- 1.2), (0.65 +/- 0.41) microg/L] were significantly lower than those in the healthy controls [(3.6 +/- 2.3), (1.03 +/- 0.71) microg/L, all P < 0.01]. The serum resistin and leptin levels in patients with malnutrition [(1.7 +/- 0.7), (0.43 +/- 0.16) microg/L] were significantly lower than those in patients without malnutrition [(2.2 +/- 1.2), (0.73 +/- 0.48) microg/L, all P < 0.05]. The serum resistin level in the patients was correlated with the serum leptin level, forced expiratory volume in one second (FEV(1)), and FEV(1)/forced vital capacity (FVC) (r = 0.426 - 0.531, all P < 0.01). The serum leptin level in the patients was correlated with serum resistin, FEV(1)/FVC, body mass index (BMI), percentage of ideal body weight (IBW%), chest circumference and abdominal circumference (r = 0.371 - 0.580, all P < 0.01). CONCLUSION The serum resistin and leptin levels in stable COPD patients were significantly lower, especially in patients with malnutrition.

A follow-up study of the lung function and the chest CT changes in medical staff with severe acute respiratory syndrome in Beijing

Abstract: Objective To analyze the lung function and radiological changes in rehabilitating severe acute respiratory syndrome(SARS) patients of medical staff in Beijing. Methods Follow-up lung function tests and chest high-resolution computerized tomography(HRCT)were performed in medical staff with SARS from Dec. 2003 to Feb. 2004. Results Thirty-one(7.64%) of 406 patients showed abnormal ventilatory function, while 165 of 404 patients showed diffusing abnormality. Of the 434 patients who had received HRCT scans,151(34.79%) showed abnormalities including subpleural and basal ground-glass and reticular attenuation,nodular septal thickening and bronchiectasis. Of the 395 patients who had received both lung function test and HRCT scanning,55(13.92%) had abnormalities both in the lung function and HRCT. The decrease in diffusing capacity in patients with HRCT changes was more significant than those without HRCT changes. Conclusions Lung function and lung imaging were abnormal in some patients with SARS after recovery. It is suggested that the lung damage is chronic,and follow-up is needed.

[Pulmonary alveolar proteinosis treated with whole-lung lavage utilizing extracorporeal membrane oxygenation: a case report and review].

Abstract: OBJECTIVE To improve the treatment of severe hypoxaemia in patients with pulmonary alveolar proteinosis (PAP). METHODS The clinical data of a patient with pathologically proven PAP treated with whole-lung lavage utilizing extracorporeal membrane oxygenation (ECMO) were described and the literature was reviewed. RESULTS This 57-year-old man was admitted because of cough and progressive dyspnea for 12 months. His PaO(2) was 46 mm Hg (1 mm Hg = 0.133 kPa) and saturation of pulse oximeter (SpO(2)) was from 85% to 88% with oxygen 5 L/min by nasal cannula. His chest CT, bronchoscopy with bronchoalveolar lavage and transbronchial biopsies were consistent with PAP. Whole-lung lavage was performed in the operation room under general anesthesia. A double-lumen tube was intubated in order to selectively ventilate and lavage a single lung independently. During mechanical ventilation for both lungs, the SpO(2) was from 80% to 90%, but when a single right lung ventilation was tried, the SpO(2) (from 68% to 80%) dropped significantly. To ensure adequate oxygen supply during lavage, a veno-arterial ECMO was set up by inserting catheters percutaneously into the right femoral artery and right femoral vein respectively. Then the SpO(2) improved, from 89% to 97% during single right lung ventilation. The left lung was lavaged with a total of 20.8 L of normal saline. The SpO(2) ranged from 80% to 94% during the lavage. After the lavage, the patient no longer experienced shortness of breath. Then 28 days later the right lung was lavaged without the aid of ECMO. A month after the second lavage, his chest CT showed marked improvement in infiltrates of both lungs. CONCLUSION When a patient with PAP has refractory hypoxemia prior to the lavage procedure, ECMO should be considered in order to avoid severe hypoxaemia with fatal consequences during lavage.

[Relationship between the acquired multi-drug resistance of human large cell lung cancer cell line NCI-H460 by cisplatin selection and p53 mutation].

Abstract: Objective To explore the role of p53 mutation in the development of acquired multi-drug resistance during lung cancer chemotherapy Methods A resistance large cell lung carcinoma cell line (H460/DDP) was established by high dose (50 micromol/L) cisplatin intermittent selection from its parental cell NCI-H460 that had wild type p53 (wtp53) Several multi-drug resistant proteins (MRP) including lung relative protein (LRP), P-gp, MRP, glutathione transferase-pi, topoisomerase II and P53 were checked by immunocytochemistry Immunofluorescence was used to check the phosphoration of P53 p53 cDNA was amplified and sequenced The H460/DDP cells were transected with plasmid pShuttle-CMV-wtp53 cDNA and measurement of drug sensitivity was performed after transfection Results The resistance index to cisplatin and carboplatin in H460/DDP cell line was 1021 and 998 respectively, and the cell line also exhibited cross-resistance to 5-fluorouracil, etoposide, methotrexate, adriamycin, epirubicin, bleomycin and novantrone, except for taxol It was found that P53 protein translocated from nuclear to cytoplasm and could not phosphorate after the stimulation of cisplatin LRP was expressed increasingly and other proteins showed insignificant changes Sequencing showed an important insertion "t" after 277 bp on the gene level H460/DDP cells transfected with plasmid pShuttle-CMV-wtp53 cDNA reversed partly (532%) the resistance to cisplatin/carboplatin, compared to the H460/DDP cells transfected with blank vector Conclusions p53 mutation induced by cisplatin/carboplatin might play an important role in the development of acquired multi-drug resistance of lung cancer chemotherapy wtp53 substitute therapy during chemotherapy may be an effective method to overcome the acquired drug resistance

[Changes of the activity and expression of gamma-glutamylcysteine synthetase in patients with chronic obstructive pulmonary disease].

Abstract: Objectie To inestigate the oxidatie/antioxidatie status in patients with chronic obstructie pulmonary disease(COPD),and to study the expression and location of γ-glutamylcysteine synthetase(γ-GCS) in lung tissues. Methods (1)Serum samples of 13 patients with COPD in exacerbation and 9 healthy non-smokers were collected for measurements of the leel of reduced glutathione(GSH),reactie oxygen species(ROS),total antioxidatie capacity(T-AOC) and γ-GCS actiity. (2)Lung tissues from 22 patients(12 patients with COPD and 10 patients as control) undergoing resection for lung tumor were collected for study of the expression and location of γ-GCS and γ-GCS mRNA by immunohistochemistry and in situ hybridization,respectiely. Results (1)Significantly decreased leel of GSH[(23.87±3.86)mg/ml],increased leel of ROS[(2 463±199)U/ml],decreased T-AOC[(5.34±0.22)U/ml] and enhanced actiity of γ-GCS[(19.22±3.36)U/ml] were shown in the serum of patients with an acute exacerbation of COPD,as compared to the control group[(36.87±6.34)mg/ml,(1 023±112)U/ml,(11.36±1.07)U/ml and (12.37±2.96)U/ml,respectiely,all P0.05]. (2)There was no relationship between the leel of GSH,ROS,T-AOC,γ-GCS actiity in serum and FE 1%,FE 1/FC,PaO 2,PaCO 2 of patients with COPD in exacerbation(P0.05). (3)In situ hybridization showed that the expressions of γ-GCS mRNA in aleoli,bronchi and inflammatory cells(A alue was 0.29±0.05,0.31±0.05 and 0.28±0.06,respectiely) from the COPD group were stronger than those from the control group(0.14±0.03,0.17±0.04 and 0.20±0.05,respectiely) by semi-quantitatie analysis(all P0.05). (4)By immunohistochemistry,the expressions of γ-GCS was significantly higher in aleoli(0.20±0.04),bronchi(0.18±0.02) and inflammatory cells(0.25±0.06) in the COPD group as compared to the control group(0.12±0.04,0.10±0.03 and 0.14±0.04,respectiely,all P0.05). (5)Negatie correlations were shown between γ-GCS and FE 1%,FE 1/FC(r=-0.501 and -0.542,respectiely,P0.05),while the leel of γ-GCS expression had no correlation with FE 1% and FE 1/FC(r=-0.221 and -0.148,respectiely,P0.05),and a positie relationship was obsered between γ-GCS and γ-GCS mRNA(r=0.732,P0.05). Conclusions Systemic oxidatie/antioxidatie imbalance occurs in patients with acute exacerbation of COPD,and the total antioxidatie capacity decrease may not correlate significantly to the obstruction of airways,in spite of the high leel of γ-GCS actiity in serum and γ-GCS expression in the lungs.

[Characteristics of the airway inflammation and the relationship to interleukin-17 in severe asthma].

Abstract: Objective To investigate the characteristics of airway inflammation in severe asthma and the association with interleukin-17(IL-17). Methods Sixteen patients with mild persistent asthma, 14 patients with moderate persistent asthma and 18 patients with severe persistent asthma, as well as 15 normal control subjects, were included in this study. At baseline, asthma symptom score was recorded, and lung function was measured. Induced sputum was obtained and processed for cell differential and the supernatant was assayed for the concentrations of IL-17 and IL-8 by sandwich enzyme-linked immunosorbent assay (ELISA), eosinophil cationic protein (ECP) by the immuno-CAP system, and neutrophil myeloperoxidase(MPO) by a colorimetric method, and the values were normalized to the protein content. Then 15 mild to moderate and 15 severe asthmatic patients received inhaled corticosteroid (Pulmicort, Astra-Zeneca) therapy for 4 weeks, and the above measurements were repeated. Results The percentage of sputum eosinophils and the level of ECP were significantly increased in mild [0.067 0±0.074 0, (155±91)×10 -6 g/g protein], moderate [0.083 0±0.044 0, (180±83)×10 -6 g/g protein] and severe [0.124 0±0.143 0, (191±87)×10 -6 g/g protein] asthmatic patients, as compared to the normal controls [0.000 0±0.001 0, (44±25)×10 -6 g/g protein, P0.01)]; but there was no significant difference among mild, moderate and severe asthma (P0.05). On the contrary, the percentage of sputum neutrophils was significantly increased in patients with severe asthma (0.589±0.203) as compared to patients with mild (0.455±0.154) and moderate (0.449±0.194) asthma and normal controls (0.313±0.134, P0.01). The level of MPO was also increased in severe asthma [(31±10 )U/g protein] as compared to mild [(12±4) U/g protein] and moderate [(22±7)U/g protein] asthma and normal controls [(10±4) U/g protein, P0.01]. The sputum level of IL-17 in severe, moderate asthma [(264±137)×10 -9 g/g protein, (172±65)×10 -9 g/g protein] was significantly higher than those in mild asthma [(126±52)×10 -9 g/g protein] and normal controls [(56±20)×10 -9 g/g protein, P0.01]. As compared to the normal controls [(83±36)×10 -9 g/g protein],the level of IL-8 in severe asthma [(531±321)×10 -9 g/g protein] was significantly increased (P0.01),and those in mild [(410±181)×10 -9 g/g protein] and moderate [(438±148)×10 -9 g/g protein] asthma were also increased, although the difference was not statistically significant (P0.05). The level of IL-17 was correlated positively with IL-8, neutrophils and MPO (r=0.525,0.349,0.602, all P0.01). After steroid therapy, the percentage of eosinophils, the levels of ECP, MPO,IL-17 and IL-8 decreased significantly in all patients combined, but the percentage of neutrophils remained unchanged (P0.05) and still significantly higher in severe asthmatic patients (0.642±0.157) as compared to mild and moderate asthmatic patients (0.394±0.147, P0.01). Conclusions Airway neutrophilia is a feature of severe asthma. The IL-17/IL-8 pathway may be involved in the initial neutrophil influx into the airways.

[Effect of doxofylline on calcium-activated potassium channels in human peripheral blood eosinophils in asthma].

Abstract: OBJECTIVE To study the effects of doxofylline on calcium-activated potassium (K(Ca)) channels of human peripheral blood eosinophils in asthma. METHODS Peripheral blood eosinophils from patients with asthma were isolated and divided equally into two groups, a control group and a doxofylline incubated group. The data were recorded using cell-attached configuration of patch-clamp technique and the kinetic changes of K(Ca) channels activated by 0.2 micromol/L platelet activating factor (PAF) were compared. RESULTS As compared with the control group, the open probability of the K(Ca) channels decreased from 0.135 +/- 0.021 to 0.044 +/- 0.018, the open time from (5.75 +/- 0.40) ms to (2.39 +/- 0.13) ms, while the close time increased from (2.17 +/- 0.50) ms to (23.73 +/- 2.50) ms in the doxofylline incubated group. The differences were significant between the two groups (all P < 0.05). CONCLUSION Doxofylline could decrease the open probability of the channels as a result of both the shortening of open period and the prolongation of close time.

[Radiological features of AIDS complicated by pulmonary tuberculosis and the association with CD4+ T lymphocytes].

Abstract: Objective To explore the imaging features in cases with AIDS complicated by pulmonary tuberculosis and the association with CD+ 4 T lymphocytes. Methods A retrospective analysis was carried out of the manifestations of chest X-rays in patients with late stage AIDS complicate by pulmonary tuberculosis (n=26 cases) and patients with pulmonary tuberculosis only (n=60 cases). The results of measurements of CD+ 4T lymphocytes were compared. Results (1) The chest X-ray features in patients with AIDS complicated by pulmonary tuberculosis showed more patchy and blurring shadows (53.8% vs 8.3%; P0.01), more military changes (23.1% vs 5.0%; P0.05), more enlarged intrathoracic lymph nodes (34.6% vs 8.3%; P0.01) as well as more extra-pulmonary tuberculous processes (23.1% vs 3.3%; P0.05) as compared to patients with pulmonary tuberculosis alone. Fewer upper lung or apical lesions (23.1% vs 76.7%; P0.01), as well as less consolidation(11.5% vs 71.7%; P0.01) and cavity formation (7.7% vs 30.0%; P0.05) were found in AIDS patients. No significant difference was found in the occurrence of hydrothorax (11.5% vs 20.0%; P0.05). (2) The relative numbers of CD+ 4 T lymphocytes in patients with AIDS complicated by pulmonary tuberculosis and in patients with pulmonary tuberculosis alone were (5.0±6.4)% and (65.3±1.5)% respectively. Atypical manifestations of tuberculosis were found in 15 out of the 26 cases of AIDS patients showing a CD+ 4 T lymphocytic count 50/μl, in 3 of the 4 cases with the count between 50/μl-100/μl , while in cases with CD+ 4T lymphocytic count between 100/μl -200/μl (n=4) and 200/μl (n=2), numbers of atypical tuberculosis were 2 and 0 respectively. Conclusions The chest X-ray changes of tuberculosis in cases with late stage AIDS were mostly of the atypical pattern, and were related to a significant decrease in CD+ 4T lymphocyte count.

[Screening and construction of recombinant BCG strains expressing the Ag85B-ESAT6 fusion protein].

Abstract: Objective To screen and construct recombinant BCG strains which express the Ag85B-ESAT6 fusion protein. Methods The heat shock protein 60 (Hsp60) and the alpha-ss signal peptide encoding sequence were amplified by PCR from Mycobacterium tuberculosis H(37)Rv and cloned into E. coli/Mycobacteria shuttle vector-pOLYG. The resulting expression vector was named pDE22, and then ag85b and esat6 genes were cloned into pDE22 at different sites. The resulting recombinant plasmids Ag85B-ESAT6 and ESAT6-Ag85 were electroporated into BCG. Positive clones were screened by hygromycin resistance and confirmed by PCR. Recombinant BCG culture supernatants were collected and analyzed by SDS-PAGE and Western blot. Results Two recombinant BCG strains were obtained, which secreted the 37,000 fusion protein in their culture supernatant, which was confirmed by Western blot with specific immune serum against Ag85B and ESAT6. Conclusions Recombinant BCG strains expressing Ag85B and ESAT6 fusion proteins of Mycobacterium tuberculosis were constructed. They may serve as new vaccine candidates for preventing tuberculosis.

Effect of protective ventilation and open lung strategy on extravascular lung water in rabbits with acute respiratory distress syndrome

Abstract: OBJECTIVE To study the effect of protective ventilation and open lung strategy on extravascular lung water index (EVLWI) in rabbits with acute respiratory distress syndrome (ARDS). METHODS Saline-lavaged, anesthetized ARDS rabbits were divided into (1) a moderate tidal volume (V(T)) zero positive end-expiratory pressure (PEEP) group (MVZP group): V(T) 12 ml/kg, PEEP 0 cm H2O; (2) a low V(T) zero PEEP group (LVZP group): V(T) 6 ml/kg, PEEP 0 cm H2O; (3) a low V(T) best PEEP group (LVBP group): V(T) 6 ml/kg, PEEP 10 cm H2O; (4) a low V(T) best PEEP + sustained inflation (SI) group (LVBP + SI group): V(T) 6 ml/kg, PEEP 10 cm H2O + SI. EVLWI was measured by single indicator thermodilution technique at baseline, 0, 1, 2 and 3 h. RESULTS In the MVZP, LVZP, LVBP, and LVBP + SI groups, EVLWI in the ARDS model [(22.3 +/- 5.6), (20.0 +/- 3.8), (25.7 +/- 9.7), (22.5 +/- 6.2) ml/kg, respectively] was significantly higher than those at baseline [(11.3 +/- 2.4), (10.2 +/- 2.4), (10.3 +/- 4.6), (9.7 +/- 2.3) ml/kg, respectively, all P < 0.05]. In the MVZP group, EVLWI at 2 h and 3 h [(32.0 +/- 12.2), (36.2 +/- 12.4) ml/kg] was higher than that of 0 h [(22.3 +/- 5.6) ml/kg, P < 0.05]. In the LVZP group EVLWI at 2 h and 3 h [(27.8 +/- 12.9), (30.3 +/- 13.0) ml/kg] was also higher than that of 0 h [(20.0 +/- 3.8) ml/kg, P < 0.05]. In the LVBP group, EVLWI at 1 h was (18.5 +/- 8.1) ml/kg and was lower than that of 0 h [(25.7 +/- 9.7) ml/kg, P = 0.027]. In the LVBP + SI group, EVLWI at 1, 2, 3 h [(16.8 +/- 6.5), (18.0 +/- 7.1), (15.7 +/- 2.7) ml/kg] was lower than that of 0 h [(22.5 +/- 6.2) ml/kg, all P < 0.05]. There was significant difference among the four groups at 1 h and 3 h (all P < 0.05). At 1 h and 3 h, compared with MVZP group, EVLWI of the LVBP and the LVBP + SI groups were significantly decreased (all P < 0.05). At 3 h, compared with the EVLWI of the LVZP group, EVLWI of the LVBP + SI group was significantly decreased (P < 0.05). CONCLUSION Lung protective ventilation and open lung strategy could decrease EVLWI.

The development and safety of cough provocation test by capsaicin inhalation

Abstract: OBJECTIVE To assess the application and the safety of capsaicin cough provocation test by dosimeter method. METHODS Capsaicin inhalation cough challenge test by dosimeter method was performed on 60 healthy volunteers (group A), 11 subjects with upper respiratory infection (group B), 10 patients with gastroesophageal reflux cough (group C) and 10 patients with asthma (group D). Each subject inhaled doubling concentrations of capsaicin (1.95, 3.90, 7.80, 15.6, 31.2, 62.5, 125, 250, 500, 1,000 micromol/L) by a breath-activated dosimeter until the concentration inducing five or more coughs (C(5)) was reached. The lg C(5) was calculated as the cough reflex sensitivity. General lung ventilation and impedance with impulse oscillometry were measured before and after the cough provocation test. RESULTS There was no serious side effect of the test in all subjects; two subjects complained of slight nausea; one of heartburn, and three of hoarseness. Before the provocation the values of FEV(1) in group A, B, C and D were (3.6 +/- 0.5) L, (3.7 +/- 0.7) L, (2.7 +/- 0.8) L and (2.1 +/- 0.8) L, compared with (3.6 +/- 0.5) L, (3.7 +/- 0.8) L, (2.6 +/- 0.7) L and (2.1 +/- 0.8) L after the test, the differences being not significant (all P > 0.05). Compared with the measurements after provocation, Zrs was (2.6 +/- 0.8) mm Hg.L(-1).s(-1) vs (2.7 +/- 0.8) mm Hg.L(-1).s(-1) in group A, (2.5 +/- 0.5) mm Hg.L(-1).s(-1) vs (2.6 +/- 0.3) mm Hg.L(-1).s(-1) in group B, (2.7 +/- 0.7) mm Hg.L(-1).s(-1) vs (2.7 +/- 0.7) mm Hg.L(-1).s(-1) in group C, (3.3 +/- 1.5) mm Hg.L(-1).s(-1) vs (3.7 +/- 2.0) mm Hg.L(-1).s(-1) in group D, the differences showed no significance (all P > 0.05 respectively). The lg C(5) value was 2.45 +/- 0.46 in group A, 2.51 +/- 0.20 in group B, 1.52 +/- 0.70 in group C, 2.34 +/- 0.56 in group D. Compared with group A, B and D, the lg C(5) value in group C was significantly different (all P 0.05). CONCLUSION Capsaicin inhalation cough provocation test by dosimeter method is a safe and specific tool for measuring cough reflex sensitivity.

Endogenous hydrogen sulfide in patients with chronic obstructive pulmonary disease

Abstract: Objective To investigate the ro le of endogenous hydrogen sulfide(H2S) in patients with chronic obstructive pulmonary disease(COPD). Methods Levels of serum H2S a nd nitric oxide(NO),lung function and cell differential count in induced sput um were studied in 27 patients with acute exacerbation of COPD(AECOPD),7 pat ients with stable COPD and 1 health subjects. Echo-Doppler assessment and art erial blood gas were measured in patients with AECOPD. Results (1)The serum H2S level was significantly higher in patients with stable COPD[(50.8±2.5)μmol/L] as compared to those in the controls[(9.8±1 .6)μmol/L] and in patients with AECOPD[(.5±2.2)μmol/L,P0.01 ]. (2)The level of serum H2S was significantly lower in smokers with AECOP D[(28.1±1.)μmol/L] as compared to nonsmokers with AECOPD[(9.4±.9 )μmol/L,P0.05] and healthy nonsmokers[(9.8±1.6)μmol/L,P 0.01]. ()There was significant difference in the serum H2S level among stable COPD patients with different severity of airway obstruction(P0.0 5);being lower in patients with stage Ⅲ[(45.1±4.1)μmol/L] as compared to stage Ⅰ obstruction[(70.2±6.2)μmol/L,P0.05]. (4)AECOPD w ith pulmonary hypertension pulmonary artery systolic pressure(PASP)≥5 mm Hg( 1 mm Hg=0.1 kPa) showed a lower serum H2S level[(26.±2.2) ,(6.2 ±2.5)μmol/L,P0.05] than that with a normal resting PASP. (5)H2 S in serum was positively correlated with NO levels(r=0.278,P=0.029 ),FEV1% predicted values(r=0.5,P=0.000),percentage of sputum lymphocytes(r=0.286,P=0.028) and macrophages (r=0.4, P=0.01);and negatively correlated with PASP(r=-0.561,P=0.011) and the percentage of sputum neutrophils(r=-0.422,P=0.001) in p atients with COPD. Conclusion Endogenous H2S may be involved in the pathogenesis of airway obstruction in COPD and may be a noninvas ive marker of disease activity and severity.

The relationship between leukemia inhibitory factor and neurokinin receptors in a rat model of asthma

Abstract: OBJECTIVE To study the expression of leukemia inhibitory factor (LIF) and neurokinin receptors (NKR) in the lungs of asthmatic rats, and to evaluate the role of LIF in airway neurogenic inflammation. METHODS Twenty-four Wistar rats were randomly divided into a control group (group A, n = 8), an asthma group (group B, n = 8) and a dexamethasone treated group (group C, n = 8). The rat asthmatic model was made by intraperitoneal injection and nebulized aspiration of ovalbumin (OVA) at the concentrations of 10% and 1% respectively. Expression levels of lung LIF, NK-1R and NK-2R were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot two weeks after challenge, and the localization of NK-1R was determined by immunohistochemistry. RESULTS After challenge, the expressions of lung LIF mRNA in group A, B and C were 0.240 +/- 0.020, 0.510 +/- 0.130, 0.180 +/- 0.050, and protein levels were 23 110 +/- 8 018, 40 832 +/- 12 964, 16 160 +/- 2 108 respectively. The expressions of lung NK-1R mRNA in group A, B and C were 0.240 +/- 0.020, 1.040 +/- 0.480, 0.170 +/- 0.040, and protein levels were 16 538 +/- 4 342, 32 292 +/- 4 564, 15 018 +/- 1 488 respectively. The mRNA and protein levels of LIF and NK-1R in group B were significantly elevated as compared with group A and C (all P 0.05). In group B, there was a positive correlation between LIF and NK-1R at mRNA (r = 0.850, P < 0.01) and protein (r = 0.868, P < 0.01) levels respectively. NK-1R immunoreactivity was observed primarily in bronchial epithelial cells. CONCLUSION LIF and NK-1R were excessively expressed and closely correlated in lungs of the rat asthmatic model, suggesting that LIF may be involved in modulating airway neurogenic inflammation.

Measurement of diaphragm compound muscle action potential with magnetic stimulation of the phrenic nerve and multipara esophageal electrode in intensive care unit

Abstract: OBJECTIVE To investigate whether magnetic stimulation of the phrenic nerve and multipara esophageal electrode can be used to measure diaphragm compound muscle action potential (CMAP) in intensive care unit (ICU). METHODS Ten normal subjects and 10 patients in ICU were studied. A newly designed multipara esophageal catheter with 5 pairs of electrodes was used to record the diaphragm CMAP elicited by magnetic stimulation and transcutaneous electrical stimulation of the phrenic nerve. RESULTS Good quality CMAPs were obtained from normal subjects and patients in ICU. In normal subjects, the phrenic nerve conduction time (PNCT) and amplitude of the diaphragm CMAP measured with electrical stimulation [(7.2 +/- 0.8) ms, (1.52 +/- 0.40) mV] were similar to those measured with magnetic stimulation [(7.1 +/- 0.8) ms, (1.56 +/- 0.38) mV, all P > 0.05, pooled left and right values]. The amplitude of the diaphragm CMAP in patients [(0.73 +/- 0.38) mV] was much smaller than that in normal subjects [(1.58 +/- 0.38) mV, P < 0.01, pooled left and right values]. CONCLUSION This study suggests that magnetic stimulation combined with a multipara esophageal electrode could be a useful technique in assessing diaphragm function in ICU.

[The effect of 20(R)-ginsenoside Rg3 on the differential expression of cell signaling genes and other related genes in human lung adenocarcinoma cell line A549].

Abstract: OBJECTIVE To study the effect of 20(R)-ginsenoside Rg3 [20(R)-Rg3] on the differential expression of cell signaling genes and other related genes in human lung adenocarcinoma cell line A549. METHODS The cell line A549 was cultured with 20(R)-Rg3 (10(-6) mol/L) for 72 h, RNA was extracted, and the differential expression of cell signaling genes and other related genes were detected with DNA microarray. RESULTS A total of 3 490 genes were detected, and the expression of 24 genes were changed after the addition of 20(R)-Rg3. Two cell signaling genes & transducin genes were up-regulated. Two cystoskeleton & locomotion genes were up-regulated. Three proto-oncogene & anti-oncogene genes were down-regulated. Two of the translation & synthetical genes were down-regulated, and 1 of them was up-regulated. One DNA recombination gene and one metabolism gene were down-regulated. CONCLUSION 20(R)-Rg3 showed significant effect on the differential expression of cell signaling genes and other related genes in human lung cell line A549.

[The role of airway neurogenic inflammation in gastro-esophageal reflux induced cough].

Abstract: OBJECTIVE To investigate the role of airway neurogenic inflammation in the pathogenesis of gastro-esophageal reflux induced cough (GERC). METHODS Sputum was induced by hypertonic saline aerosol inhalation in 20 patients with GERC (GERC group), 10 healthy subjects (normal control group) and 8 patients with chronic cough due to other causes but complicated with gastro-esophageal reflux diseases (GERD, GERD group). Airway mucosal biopsy was performed in 6 patients with GERC and 4 patients with GERD using flexible fiberoptic bronchoscopy. The expression of substance P (SP), neurokinin 1 receptor and neurokinin A (NKA) in sputum cells and airway mucosa were detected by immunohistochemistry, and was assessed semi-quantitatively. SP, NKA, and NKB in the supernatant of induced sputum were measured with enzyme linked immunosorbent assay. Calcitonin gene-related peptide (CGRP) was measured with radioimmunoassay. RESULTS The concentration of SP in the supernatant of induced sputum was significantly higher in GERC group [(266 +/- 207) ng/L] than those in normal control group [(143 +/- 36) ng/L, P < 0.05] and GERD group [(130 +/- 11) ng/L, P < 0.05], and the sputum supernatant concentration of CGRP in GERC group [(180 +/- 83) ng/L] was significantly higher than those in normal control group [(105 +/- 64) ng/L, P < 0.01] and GERD group [(89 +/- 16) ng/L, P < 0.01]. The expression of SP, NK-1 receptor and NKA in induced sputum cells in GERC group were significantly higher than those in normal control group (P < 0.01, < 0.05, < 0.05) and GERD group (all P < 0.05); Expressions of SP in airway mucosa was significantly higher in GERC group than in GERD group (P < 0.01). After treatment, the concentration of CGRP in the supernatant of sputum in GERC patients was significantly lower than that before treatment (P < 0.05); the expression of SP, NK-1 and NKA in the induced sputum cells were significantly lower than that before treatment (P < 0.01, P < 0.01 or P < 0.05). CONCLUSION There is airway neurogenic inflammation in GERC patients, which maybe closely related to the development of GERC.

[Experimental study on the expression and function of aquaporin-1 and aquaporin-5 in rats with acute lung injury induced by lipopolysaccharide].

Abstract: OBJECTIVE To investigate whether the expression and function of aquaporin-1 (AQP-1) is altered by tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in primary rat lung microvessel endothelial cells (LMECs) after exposure to lipopolysaccharide (LPS), and to study the expressions of AQP-1 and AQP-5 in lung tissue of rats with acute lung injury (ALI) induced by LPS. The aim is to further clarify the pathogenesis of ALI/acute respiratory distress syndrome (ARDS). METHODS (1) In vitro: The third passage LMECs were randomly divided into LPS group, TNF-alpha group, IL-1beta group and DMEM control group, and the experimental groups were exposed to LPS, TNF-alpha and IL-1beta respectively. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to quantify AQP-1 mRNA changes and an immunocytochemistry method was used for determining AQP-1 protein changes in cultured rat LMECs. Isotope tracer technique was applied for the assay of the intra-cellular tritium water ((3)H2O) signal intensity in rat LMECs. (2) In vivo: Forty male Wistar rats were randomly divided into five groups: LPS 2 h group, LPS 4 h group, LPS 6 h group, LPS 8 h group and a control group, eight rats per group; The LPS treated groups served as the ALI models. RT-PCR was used to observe the changes of AQP-1 and AQP-5 mRNA and the immunohistochemistry method was used for determining AQP-1 and AQP-5 protein changes in ALI rats. RESULTS (1) In vitro: The expression of AQP-1 mRNA and protein in LMECs were decreased significantly in the LPS group (0.428 +/- 0.026, 0.366 +/- 0.009), the TNF-alpha group (0.446 +/- 0.029, 0.374 +/- 0.014) and IL-1beta group (0.454 +/- 0.023, 0.377 +/- 0.007) as compared to the DMEM control group (0.793 +/- 0.035, 0.660 +/- 0.013, respectively; all P < 0.01). The quantities of tritium water's permeability in the LPS group, the TNF-alpha group and the IL-1beta group [(726 +/- 58), (738 +/- 45), (774 +/- 44) counts per minute] were significantly less than that in the DMEM control group [(1 148 +/- 70) counts per minute, P < 0.01]. (2) In vivo: The expression levels of AQP-1 and AQP-5 mRNA in ALI rats (LPS 2 h group 0.409 +/- 0.018, 0.421 +/- 0.020; LPS 4 h group 0.421 +/- 0.023, 0.412 +/- 0.023; LPS 6 h group 0.435 +/- 0.020, 0.388 +/- 0.031; LPS 8 h group 0.438 +/- 0.016, 0.386 +/- 0.019, respectively) were significantly lower than that in the control group (0.794 +/- 0.015, 0.787 +/- 0.022; all P < 0.01). CONCLUSION AQP-1 and AQP-5 may play a role in abnormal fluid transportation and probably involve in the formation of pulmonary edema in ALI/ARDS.

The roles of dendritic cells in antigen presentation and the pathogenesis of asthma

Abstract: OBJECTIVE To investigate the roles of dendritic cells (DCs) in the pathogenesis of asthma. METHODS (1) DCs were derived from peripheral blood monocytes of asthmatics with acute exacerbation (group A, n = 20), patients at remission (group B, n = 15) and healthy volunteers (group C, n = 10), and cultured using media supplemented with granulocyte-macrophage colony stimulation factors (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha). The antigen-uptaking function of the monocyte-derived DCs was evaluated by phagocytosis of fluorescence labeled ovalbumin (OVA), and their antigen-presenting function was determined by expression of membrane markers (MHC-II) and co-stimulatory molecules (CD(80) and CD(86)). The proliferation of T lymphocytes induced by DCs was assessed using autologous mixed T lymphocyte reaction. (2) The expression of MHC-II and CD(80) and CD(86) of DCs were determined from the bronchoalveolar lavage (BAL) of asthmatic rats. The effects of dexamethasone on the expression of these markers and molecules were studied. Twenty-four rats were divided into the experimental group (group D) and the control group (group E). In group D, rats were subdivided into two groups with 8 rats in each. One group (group D(2)) received pre-treatment with dexamethasone (10 mg/kg), and another group (group D(1)) did not received dexamethasone pre-treatment, while group E was matched with the experimental group with regard to both age and weight. The rats were sensitized and challenged with OVA, and BAL was obtained. (3) All the membrane markers and co-stimulatory molecules on the surface of DCs were determined using flow cytometric method. RESULTS (1) The phagocytosis of fluorescence labeled OVA in group A was increased [(41 +/- 12)%] compared with those in group B [(29 +/- 10)%, P < 0.01] and group C [(29 +/- 10)%, P < 0.01]. The expression of MHC-II, CD(1alpha) and CD(80) on the surface of DCs was the highest in group A [(44 +/- 15)%, (32 +/- 11)% and (32 +/- 13)%, respectively] compared with those in group B [(22 +/- 10)%, (19 +/- 9)% and (19 +/- 10)%, all P < 0.01, respectively], as well as with those in group C [(18 +/- 12)%, (13 +/- 7)% and (14 +/- 7)%, all P < 0.01, respectively]. There was no statistical difference in the expression of CD(86) on the cell membrane in group A, group B and group C. In autologous mixed T lymphocyte reaction study, when DC/T ratio was 1/5, DCs had the highest ability in stimulating the proliferation of T lymphocytes in group A (stimulating index = 2.32 +/- 0.44) compared with those in group B (1.01 +/- 0.11, P < 0.01), and those in group C (1.62 +/- 0.27, P < 0.01). (2) The expression of MHC-II in BAL cells of asthmatic rats reached peak value at 6 h after challenge, (15.2 +/- 5.0)% in group D(1), and (2.0 +/- 1.0)% in group E, P < 0.01. The expression of CD(80) and CD(86) increased rapidly 2 h after challenge in group D(1) [(10.6 +/- 3.9)% and (7.5 +/- 3.8)%, respectively] compared with those in group E [(2.1 +/- 0.7)%, P < 0.01 and (1.7 +/- 0.7)%, P < 0.05, respectively]. The maximal inhibition effect of dexamethasone on the expression of MHC-II was at 10 h, (7.8 +/- 2.4)% in group D(1), and (2.8 +/- 1.5)% in group D(2), P < 0.01. The inhibition effect of dexamethasone on the expression of CD(80) and CD(86) was maximal at 24 h, (5.8 +/- 2.7)% and (5.5 +/- 1.5)% respectively in group D(1), and (2.8 +/- 1.1)% and (2.9 +/- 1.6)% respectively in group D(2), P < 0.05. CONCLUSIONS The antigen up-taking and presenting function of DCs were significantly enhanced in asthmatics at exacerbation and in a rat asthmatic model. Pretreatment with dexamethasone inhibited the function of DCs significantly in the animal model. The results suggest that DCs play important roles in antigen presentation in the pathogenesis of asthma, and the inhibition of glucocorticiod on DCs might be a critical mechanism in treatment of bronchial asthma.

[The preventive effect of bacillus Calmette-Guerin vaccination in early life on airway inflammation and mucus production in a murine model of asthma].

Abstract: OBJECTIVE To investigate the preventive effect of bacillus Calmette-Guerin (BCG) vaccination in early life on airway inflammation and mucus production in a murine model of asthma. METHODS Twenty-five newborn C57BL/6 mice were divided into three groups including group A (saline sensitized/challenged mice, n = 8), group B [ovalbumin (OVA) sensitized/challenged mice, n = 9], group C (OVA sensitized/challenged mice treated by BCG in early life, n = 8). Mice in group C were subcutaneously injected by 25 microl BCG (10(3) CFU) four times at 0 week, 1st week, 2nd week and 3rd week of age respectively, while mice in group A and B were injected by the same volume of saline solution. Mice were sensitized on day 42 and 56 by saline (group A) or OVA (group B and C) and challenged from day 66 to 68 by saline (group A) or OVA (group B and C) repeatedly. Samples were collected at 48 h after the last OVA challenge. The outcome measurements included eosinophil count in bronchoalveolar lavage fluid (BALF), periodic acid-schiff reaction (PAS) staining of lung tissues, the goblet cell hyperplasia ratio (HR) and the epithelial cell mucus occupying ratio (MOR). The mRNA expression of mucus gene (MUC5AC) was measured by reverse trancription-polymerase chain reaction (RT-PCR). RESULTS The total cells in BALF in group B (123.8 +/- 14.5) x 10(4)/ml] and that in group C [(77.4 +/- 14.1) x 10(4)/ml] were increased significantly as compared with that in group A [(15.0 +/- 1.4) x 10(4)/ml, P 0.05). The number of eosinophils in BALF in group B [(78.220 +/- 11.770) x 10(4)/ml] and that in group C [(24.430 +/- 7.020) x 10(4)/ml] were increased significantly when compared with that in group A [(0.090 +/- 0.020) x 10(4)/ml, P < 0.01, P < 0.05], and that in group C was significantly reduced when compared with that in group B (P < 0.01). The HR and MOR in group B [(41.80 +/- 4.09)/mm, (28.43 +/- 2.75)% respectively] and those in group C [(21.58 +/- 5.09)/mm, (9.43 +/- 2.30)% respectively] were increased significantly when compared with those in group A [(3.00 +/- 1.20)/mm, (0.75 +/- 0.29)% respectively, P < 0.01, P < 0.01], but those in group C were significantly reduced when compared with those in group B (P < 0.01). The mRNA expression of MUC5AC in group B (1.07 +/- 0.03) and that in group C (0.86 +/- 0.05) were increased significantly when compared with that in group A (0.64 +/- 0.08, P < 0.01, P < 0.01), but that in group C were significantly reduced when compared with that in group B (P < 0.01). CONCLUSION BCG vaccination in early life can inhibit the development of airway inflammation and mucus production in a murine model of asthma.

[Effects of valsartan on bleomycin-induced pulmonary fibrosis in rats and the expression of hepatocyte growth factor in lung tissue].

Abstract: OBJECTIVE To study the effect of valsartan on bleomycin-induced pulmonary fibrosis in rats and its possible mechanism. METHODS Sixty Wistar rats were divided into three groups. In the valsartan group and the model groups, pulmonary fibrosis was induced by intratracheal instillation of bleomycin (BLM), and then the former group received valsartan 16 mg/kg daily, but the latter group received oral normal saline. In the control group, normal saline was given both intratracheally and orally. Five rats in each group were sacrificed 7, 14, 28, and 42 days after intratracheal instillation. Collagen content of the lung tissue was assessed by hydroxyproline concentration. Histological changes of the lungs were evaluated by HE stain and Masson's trichrome stain. Lung expression of transforming growth factor-beta (TGF-beta) protein was assessed by immunohistochemistry. The mRNA of hepatocyte growth factor (HGF) was detected by in situ hybridization, and the level of HGF protein was further measured by ELISA. RESULTS In the valsartan group, alveolitis [(0.88 +/- 0.12) points, (0.79 +/- 0.21) points, (0.75 +/- 0.17) points] decreased significantly 14 days after treatment with BLM and was significantly lower as compared with the model group [(2.05 +/- 0.25) points, (1.38 +/- 0.12) points, (1.19 +/- 0.11) points, P < 0.01]. Pulmonary fibrosis on day 7, 28, 42 [(0.28 +/- 0.03) points vs (0.45 +/- 0.10) points, (1.74 +/- 0.18) points vs (2.08 +/- 0.32) points, (1.91 +/- 0.09) points vs (2.77 +/- 0.15) points, P < 0.05], hydroxyproline concentrations on day 14, 42 [(1.08 +/- 0.13) microg/mg x pro vs (1.45 +/- 0.19) microg/mg x pro, (1.39 +/- 0.20) microg/mg x pro vs (2.19 +/- 0.37) microg/mg x pro, P < 0.01] and protein expression of TGF-beta on day 7, 42 [2.27 +/- 0.30 vs 3.99 +/- 0.43, 2.05 +/- 0.18 vs 2.71 +/- 0.46, P < 0.05] in the valsartan group all reduced significantly as compared with that of the model group. HGF mRNA [1.61 +/- 0.20 vs 1.98 +/- 0.23, 0.52 +/- 0.19 vs 0.28 +/- 0.14, P < 0.05] and its protein expression (204 +/- 18) pg/ml vs (260 +/- 21) pg/ml, (129 +/- 20) pg/ml vs (100 +/- 20) pg/ml, P < 0.05) on day 7 and day 42 in the valsartan group were increased as compared with those of the model group. CONCLUSIONS Valsartan alleviates BLM-induced pulmonary fibrosis in rats. Inhibiting the expressions of TGF-beta and stimulating the expression of HGF in lung tissues may be one of the mechanisms.

[The interleukin-6 level in exhaled breath condensate of patients with obstructive sleep apnea-hypopnea syndrome].

Abstract: Objective To explore whether the airway inflammation marker in exhaled breath condensate is increased in obstructive sleep apnea-hypopnea syndrome(OSAHS) Methods Thirty-one patients with OSAHS(15 smokers and 16 non-smokers) and 10 healthy age- matched and weight-matched controls were included in the study Exhaled breath condensate(EBC) was collected before and after sleep at the same night from both groups Interleukin-6(IL-6) in EBC was measured by a specific enzyme immunoassay Results (1)There was no difference in the pre-sleep IL-6 level among OSAHS smoker group,OSAHS non-smoker group and the control subjects(F=0515,P005) Compared with the level of pre-sleep[(25±10)ng/L in OSAHS smoker,and (23±08)ng/L in OSAHS non-smoker],the post-sleep level of IL-6 was elevated significantly in EBC from OSAHS patients of non-smokers[(31±12)(ng/L)] and smokers[(37±19)ng/L,P005] Nevertheless,IL-6 level from the control group showed a reverse change IL-6 was decreased significantly[(20±08)ng/L in pre-sleep vs (27±10)ng/L in post-sleep] after sleep in this group There was no difference in post-sleep IL-6 level between OSAHS smokers[(37±19)ng/L] and non-smokers[(31±12)ng/L,P005] A significant higher IL-6 level was observed in both OSAHS smokers[(37±19)ng/L] and non-smokers[(31±12)ng/L] compared with the controlled group[(20±08)ng/L,P005] IL-6 level in EBC was correlated positively with AHI(r=0441, P005),ODI4(r=0533, P005),and negatively with minimal oxygen saturation(r=(-0529), P005) Conclusions These findings suggest that inflammation was characteristic in the airways of OSAHS patients Nocturnal hypoxia could be responsible for this change The levels of IL-6 in EBC are associated with the severity of OSAHS and may prove to be useful in monitoring of airway inflammation in OSAHS