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1.
Braz J Med Biol Res ; 54(4): e9850, 2021.
Article in English | MEDLINE | ID: mdl-33656056

ABSTRACT

Respiratory syncytial virus (RSV) infection is the main cause of lower respiratory tract infection in children. However, there is no effective treatment for RSV infection. Here, we aimed to identify potential biomarkers to aid in the treatment of RSV infection. Children in the acute and convalescence phases of RSV infection were recruited and proteomic analysis was performed to identify differentially expressed proteins (DEPs). Subsequently, promising candidate proteins were determined by functional enrichment and protein-protein interaction network analysis, and underwent further validation by western blot both in clinical and mouse model samples. Among the 79 DEPs identified in RSV patient samples, 4 proteins (BPGM, TPI1, PRDX2, and CFL1) were confirmed to be significantly upregulated during RSV infection. Functional analysis showed that BPGM and TPI1 were mainly involved in glycolysis, indicating an association between RSV infection and the glycolysis metabolic pathway. Our findings provide insights into the proteomic profile during RSV infection and indicated that BPGM, TPI1, PRDX2, and CFL1 may be potential therapeutic biomarkers or targets for the treatment of RSV infection.


Subject(s)
Respiratory Syncytial Virus Infections , Respiratory Syncytial Virus, Human , Biomarkers , Child , Humans , Proteomics
2.
Nan Fang Yi Ke Da Xue Xue Bao ; 37(10): 1301-1307, 2017 Oct 20.
Article in Chinese | MEDLINE | ID: mdl-29070458

ABSTRACT

OBJECTIVE: To explore the role of the receptor for advanced glycation end products (RAGE) in regulating the expression of MUC5AC and mucus production in a mouse model of toluene diisocyanate (TDI)?induced asthma. METHODS: BALB/c mice were randomly divided into control group, vehicle (AOO) group, TDI?induced asthma group and RAGE inhibitor (FPS?ZM1) group. PAS staining, Western blotting, and immunohistochemistry were used to analyze the changes in mucus production and MUC5AC expression in the airway of the mice, and the expression of p?ERK was detected with Western blotting. In vitro cultured human bronchial epithelial cell line 16HBE was transfected with lentiviral vector carrying short hairpin RNA targeting RAGE (shRNA?RAGE) and subsequently challenged with a TDI?human serum albumin (TDI-HSA) conjugate, and the changes in cellular MUC5AC mRNA expression as detected using RT-PCR; the protein expressions of ERK and p?ERK in the cells were examined with Western blotting. The effect of ERK inhibitor U0126 pretreatment on MUC5AC mRNA expression was also analyzed in the cells. RESULTS: Compared with the control mice, TDI-induced asthmatic mice showed significantly higher rates of PAS positivity and increased MUC5AC and p?ERK expressions in the airway (P<0.05). Treatment with FPS?ZM1 significantly decreased PAS positivity and lowered MUC5AC and p?ERK expressions in the airway of the asthmatic mice (P<0.05). Exposure of 16HBE cells to TDI?HSA caused a significant increase in MUC5AC mRNA expression and p?ERK protein expression (P<0.05), while RAGE knockdown obviously suppressed TDI?HSA-induced upregulation of p-ERK and MUC5AC mRNA (P<0.05). Treatment with the ERK inhibitor U0126 also lowered TDI?HSA?induced up?regulation of MUC5AC mRNA in the cells (P<0.05). CONCLUSION: RAGE signaling induces MUC5AC expression via extracellular signal-regulated kinase pathway to promote mucus overproduction in mice with TDI-induced asthma.


Subject(s)
Asthma/metabolism , Mucin 5AC/metabolism , Mucus/metabolism , Receptor for Advanced Glycation End Products/metabolism , Animals , Asthma/chemically induced , Benzamides/pharmacology , Butadienes/pharmacology , Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors , Mice , Mice, Inbred BALB C , Nitriles/pharmacology , Random Allocation , Receptor for Advanced Glycation End Products/antagonists & inhibitors , Toluene 2,4-Diisocyanate
3.
Nan Fang Yi Ke Da Xue Xue Bao ; 37(6): 737-743, 2017 Jun 20.
Article in Chinese | MEDLINE | ID: mdl-28669945

ABSTRACT

OBJECTIVE: To investigate the role of epidermal growth factor receptor (EGFR) signaling pathway in bronchial epithelial actin stress fiber (F-actin) rearrangement induced by house dust mite (HDM). METHODS: Normal human bronchial epithelial cells (16HBE) were stimulated with HDM with or without pretreatment with AG-1478, an EGFR inhibitor. The levels of phospho(p)-EGFR, F-actin, E-cadherin and ß-catenin in the cell cultures were detected with Western blotting. The localizations of F-actin, E-cadherin and ß-catenin in the bronchial epithelial cells were determined with immunofluorescence assay, and the transmembrane electrical resistance (TER) and FITC-dextran flux (FITC-DX) in the cells were measured to assess the barrier function of the bronchial epithelia. RESULTS: HDM stimulation of the cells for 10 min resulted in significantly increased p-EGFR expression (P<0.05) without causing obvious changes in the expression of E-cadherin (P>0.05) or ß-catenin (P>0.05). Immunofluorescence assay revealed delocalization of E-cadherin and ß-catenin in HDM-treated 16HBE cells, shown by their diffusion from the cell membrane to the cytoplasm. In HDM-treated cells, the TER was significantly decreased to (70.00∓4.33)% and the FITC-DX was significantly increased to (115.98∓4.34)%; Inhibition of EGFR reversed the delocalization of E-cadherin and ß-catenin, improved the TER to (90.00∓3.75)% and lowered the FITC-DX to (101.10∓2.10)%. HDM induced increased expression and rearrangement of F-actin, which was obviously inhibited by pretreatment of the cells with AG-1478 (P<0.05). CONCLUSION: EGFR signaling pathway mediates HDM-induced F-actin rearrangement in human bronchial epithelial cells to contribute to epithelial barrier dysfunction.


Subject(s)
Epithelial Cells/cytology , ErbB Receptors/metabolism , Pyroglyphidae , Respiratory Mucosa/physiopathology , Signal Transduction , Actins/metabolism , Animals , Antigens, CD/metabolism , Bronchi/cytology , Cadherins/metabolism , Cells, Cultured , Humans , beta Catenin/metabolism
4.
Respir Res ; 18(1): 111, 2017 05 30.
Article in English | MEDLINE | ID: mdl-28558721

ABSTRACT

BACKGROUND: The disruption and hyperpermeability of bronchial epithelial barrier are closely related to the pathogenesis of asthma. House dust mite (HDM), one of the most important allergens, could increase the airway epithelial permeability. Heat shock protein (Hsp) 90α is also implicated in the lung endothelial barrier dysfunction by disrupting RhoA signaling. However, the effect of extracellular Hsp90α (eHsp90α) on the bronchial epithelial barrier disruption induced by HDM has never been reported. METHODS: To investigate the involvement of eHsp90α in the bronchial epithelial barrier disruption induced by HDM, normal human bronchial epithelial cell line 16HBE14o- (16HBE) cells were treated by HDM, human recombinant (hr) Hsp90α and hrHsp90ß respectively and pretreated by1G6-D7, a specific anti-secreted Hsp90α monoclonal antibody (mAb). Hsp90α-silencing cells were also constructed. To further evaluate the role of RhoA signaling in this process, cells were pretreated by inhibitors of Rho kinase, GSK429286A and Y27632 2HCl. Transepithelial electrical resistance (TEER) and FITC-dextran flux (FITC-DX) were examined as the epithelial barrier function. Expression and localization of adherens junctional proteins E-cadherin and ß-catenin were evaluated by western blotting and immunofluorescence respectively. The level of eHsp90α was investigated by concentration and purification of condition media. RhoA activity was determined by using a Rho G-LISA® RhoA activation assay kitTM biochem kit, and the phosphorylation of myosin light chain (MLC), the downstream signal molecule of RhoA, was assessed by western blotting. RESULTS: The epithelial barrier disruption and the loss of adherens junctional proteins E-cadherin and ß-catenin in cytomembrane were observed in HDM-treated 16HBE cells, paralleled with the increase of eHsp90α secretion. All of which were rescued in Hsp90α-silencing cells or by pretreating 16HBE cells with 1G6-D7. Also, 1G6-D7 suppressed RhoA activity and MLC phosphorylation induced by HDM. Furthermore, inhibitors of Rho kinase prevented and restored the airway barrier disruption. Consistently, it was hrHsp90α instead of hrHsp90ß that promoted barrier dysfunction and activated RhoA/MLC signaling in 16HBE cells. CONCLUSIONS: The eHsp90α mediates HDM-induced human bronchial epithelial barrier dysfunction by activating RhoA/MLC signaling, suggesting that eHsp90α is a potential therapeutic target for treatment of asthma.


Subject(s)
Anti-Asthmatic Agents/pharmacology , Bronchi/drug effects , Epithelial Cells/drug effects , HSP90 Heat-Shock Proteins/pharmacology , Myosin Light Chains/metabolism , Pyroglyphidae/immunology , Signal Transduction/drug effects , rhoA GTP-Binding Protein/metabolism , Animals , Antigens, CD , Bronchi/enzymology , Bronchi/immunology , Cadherins/metabolism , Cell Line , Dextrans/metabolism , Electric Impedance , Epithelial Cells/enzymology , Epithelial Cells/immunology , Fluorescein-5-isothiocyanate/analogs & derivatives , Fluorescein-5-isothiocyanate/metabolism , HSP90 Heat-Shock Proteins/genetics , HSP90 Heat-Shock Proteins/metabolism , Humans , Permeability , Phosphorylation , RNA Interference , Time Factors , Transfection , beta Catenin/metabolism , rho-Associated Kinases/metabolism
5.
Nan Fang Yi Ke Da Xue Xue Bao ; 36(7): 932-5, 2016 Jun 20.
Article in Chinese | MEDLINE | ID: mdl-27435771

ABSTRACT

OBJECTIVE: To evaluate fractional exhaled nitric oxide (FENO) level in patients with subacute cough and its value in predicting the patients' response to inhaled corticosteroids (ICS) treatment. METHODS: A total of 100 patients with persistent cough lasting more than 3 weeks were enrolled, including 52 patients with subacute cough and 48 with chronic cough. FENO, spirometry, and responses to ICS therapy of the patients were evaluated. RESULTS: The recruited patients had a median (inter-quartile ranges) FENO level of 19 ppb (12-30 ppb). Patients with chronic cough had a significantly higher median FENO level than those with subacute cough (20.5 vs 16 ppb; Z=-2.245, P=0.025). A FENO level ≥25 ppb was recorded in 15 (28.8%) patients with subacute cough, as compared with 20 (41.6%) in patients with chronic cough (χ(2)=1.801, P=0.179). With a FENO ≥25 ppb as the critical value to justify ICS treatment, 15 patients with subacute cough received ICS and 14 (93.3%) of them showed obvious relief of cough after 2 weeks of therapy, a response rate similar to that of 85.0% (17/20) in patients with chronic cough receiving the treatment (χ(2)=0.588, P=0.443). In patients with subacute cough, those with cough variant asthma (CVA) or eosinophilic bronchitis (EB) had a significantly higher median FENO level than those with postinfectious cough [(16 (11-31) ppb vs 11 (8-19) ppb, P<0.01]. In the etiological analysis, CVA or EB was identified in 23 (44.2%) of the patients with subacute cough, as compared 21 (43.8%) in patients with chronic cough (χ(2)=0.002, P=0.961). CONCLUSION: FENO may be an important indicator for etiological diagnosis of subacute cough and for predicting the response to ICS treatment.


Subject(s)
Adrenal Cortex Hormones/therapeutic use , Cough/drug therapy , Nitric Oxide/analysis , Breath Tests , Chronic Disease , Cough/diagnosis , Exhalation , Female , Humans , Male
6.
Atherosclerosis ; 243(2): 469-76, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26520902

ABSTRACT

BACKGROUND: Evidence describing the association between pulmonary function and carotid atherosclerosis has been inconclusive and the role of smoking in this association is unclear. We therefore examined this association in the Guangzhou Biobank Cohort Study-CVD Subcohort. METHODS: Common carotid artery (CCA) intima-media thickness (IMT) and carotid plaques were measured by B-mode ultrasonography and lung function by spirometry using a turbine flowmeter. Chronic obstructive pulmonary disease (COPD) was defined as the ratio of forced expiratory volume in 1 s (FEV1) to forced vital capacity (FVC) of less than 0.70. Predicted FEV1 and FVC were derived using equations for Chinese. RESULTS: Of 1625 participants aged 50 + years, 382 (23.5%) had evidence of carotid plaque. The mean CCA-IMT was higher in those with COPD than those without (0.82 ± 0.29 mm versus 0.76 ± 0.31 mm, P = 0.02). We found no evidence that the association of pulmonary function with CCA-IMT varied by smoking status (P values interaction: 0.23-0.83). After adjustment for a wide range of potential confounders, the increased risks of thickened CCA-IMT (CCA-IMT ≥1.0 mm) in those with COPD became marginally nonsignificant (adjusted odds ratio (OR) 1.45, 95% confidence interval (CI) 0.91-2.29; P = 0.12). Compared to those in the highest tertile, participants in the lowest tertile of FEV1 observed to predicted ratio had increased risk of thickened CCA-IMT (adjusted OR 2.18, 95% CI 1.42-3.34) and carotid plaque (adjusted OR 1.50, 95% CI 1.08-2.09), while participants in the lowest tertile of FVC observed to predicted ratio had increased risk of thickened CCA-IMT (adjusted OR 2.29, 95% CI 1.46-3.58), but the adjusted OR for carotid plaque was marginally nonsignificant (adjusted OR 1.29, 95% CI 0.93-1.80; P = 0.13). CONCLUSION: Independent of smoking status, poor pulmonary function was dose-dependently associated with carotid atherosclerosis in older Chinese. (281 words).


Subject(s)
Carotid Artery Diseases/physiopathology , Lung/physiopathology , Pulmonary Disease, Chronic Obstructive/physiopathology , Age Factors , Aged , Biological Specimen Banks , Carotid Artery Diseases/diagnosis , Carotid Artery Diseases/epidemiology , Carotid Artery, Common/diagnostic imaging , Carotid Intima-Media Thickness , Chi-Square Distribution , China/epidemiology , Cohort Studies , Cross-Sectional Studies , England , Female , Forced Expiratory Volume , Humans , Logistic Models , Male , Middle Aged , Multivariate Analysis , Odds Ratio , Predictive Value of Tests , Pulmonary Disease, Chronic Obstructive/diagnosis , Pulmonary Disease, Chronic Obstructive/epidemiology , Risk Assessment , Risk Factors , Spirometry , Vital Capacity
7.
Medicine (Baltimore) ; 94(33): e1263, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26287412

ABSTRACT

The strong association between bcl-2-like 11 (BIM) triggered apoptosis and the presence of epidermal growth factor receptor (EGFR) mutations has been proven in nonsmall cell lung cancer (NSCLC). However, the relationship between EGFR-tyrosine kinase inhibitor's (TKI's) efficacy and BIM polymorphism in NSCLC EGFR is still unclear.Electronic databases were searched for eligible literatures. Data on objective response rates (ORRs), disease control rates (DCRs), and progression-free survival (PFS) stratified by BIM polymorphism status were extracted and synthesized based on random-effect model. Subgroup and sensitivity analyses were conducted.A total of 6 studies that involved a total of 773 EGFR mutant advanced NSCLC patients after EGFR-TKI treatment were included. In overall, non-BIM polymorphism patients were associated with significant prolonged PFS (hazard ratio 0.63, 0.47-0.83, P = 0.001) compared to patients with BIM polymorphism. However, only marginal improvements without statistical significance in ORR (odds ratio [OR] 1.71, 0.91-3.24, P = 0.097) and DCR (OR 1.56, 0.85-2.89, P = 0.153) were observed. Subgroup analyses showed that the benefits of PFS in non-BIM polymorphism group were predominantly presented in pooled results of studies involving chemotherapy-naive and the others, and retrospective studies. Additionally, we failed to observe any significant benefit from patients without BIM polymorphism in every subgroup for ORR and DCR.For advanced NSCLC EGFR mutant patients, non-BIM polymorphism ones are associated with longer PFS than those with BIM polymorphism after EGFR-TKIs treatment. BIM polymorphism status should be considered an essential factor in studies regarding EGFR-targeted agents toward EGFR mutant patients.


Subject(s)
Apoptosis Regulatory Proteins/genetics , Carcinoma, Non-Small-Cell Lung , Genes, erbB-1/genetics , Membrane Proteins/genetics , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins/genetics , Apoptosis/drug effects , Apoptosis/genetics , Bcl-2-Like Protein 11 , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/metabolism , Drug Resistance, Neoplasm/genetics , ErbB Receptors/antagonists & inhibitors , Humans , Mutation , Polymorphism, Genetic , Treatment Outcome
8.
Zhonghua Yi Xue Za Zhi ; 93(18): 1374-7, 2013 May 14.
Article in Chinese | MEDLINE | ID: mdl-24025500

ABSTRACT

OBJECTIVE: To explore the risk factors for hospitalization case fatality of patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: A retrospective review of medical records was performed for 182 hospitalized AECOPD patients at Nanfang Hospital from January 2010 to August 2012. Their general information, condition in stable stage, the results of spirometry, blood routine test, blood gas analysis and C-reactive protein (CRP) were collected and analyzed. The risk factors for mortality were analyzed by multivariable Logistic regression. RESULTS: Among them, 42 died during hospitalization. Univariate analysis revealed that 8 factors had significant differences between two groups (all P < 0.05): high exacerbation risk (death vs improvement group, 90.4% vs 70.0%) , low peripheral absolute lymphocyte count (73.8% vs 47.1%), high CRP (50.0% vs 17.1%), concurrent anemia (50.0% vs 27.1%), hypoproteinemia (71.4% vs 46.4%), hypercapnia (64.3% vs 30.7%), chronic pulmonary heart disease (76.1% vs 40.7%) and ischemic heart disease (19.0% vs 7.0%). By multiple Logistic regression analysis, high CRP (OR = 3.226, P = 0.009), hypercapnia (OR = 2.928, P = 0.013), chronic pulmonary heart disease (OR = 2.510, P = 0.045), low peripheral absolute lymphocyte count (OR = 2.488, P = 0.045) were the independent risk factors for hospitalization case fatality of AECOPD patients. CONCLUSION: Low peripheral absolute lymphocyte count, high CRP, hypercapnia and chronic pulmonary heart disease were the independent risk factors for mortality in hospitalized AECOPD patients.


Subject(s)
Hospital Mortality , Pulmonary Disease, Chronic Obstructive/mortality , Aged , Female , Humans , Logistic Models , Male , Retrospective Studies , Risk Factors , Severity of Illness Index
9.
COPD ; 10(6): 650-6, 2013 Dec.
Article in English | MEDLINE | ID: mdl-23845033

ABSTRACT

Previous studies have shown that matrix metalloproteinase-9 (MMP-9) and its cognate inhibitor TIMP-1, inflammatory cytokine TNF-α, and the OPG/RANK/RANKL system may each play individual roles in the pathogenesis of osteoporosis in patients with COPD. In the present study, we investigated the interrelationships of these factors in male COPD patients with and without osteoporosis. The serum levels of MMP-9, MMP-9/TIMP-1 ratio, TNF-α, RANKL, OPG, and the RANKL/OPG ratio were higher in COPD patients with osteoporosis than in individuals with normal or low bone mineral density (BMD) (N = 30, all P < 0.05 or < 0.01). The lung function FEV1%Pre and the BMD of the lumbar spine and femoral neck were found to be negatively correlated with MMP-9 serum level (r = -0.36, P < 0.05, r = -0.58, P < 0.001, and r = -0.62, P < 0.01, respectively), RANKL serum level (r = -0.21, P < 0.05, and r = -0.25, P < 0.05, and r = -0.26, P < 0.05, respectively), and RANKL/OPG ratio (r = -0.23, P < 0.05, r = -0.33, P < 0.05, and r = -0.38, P < 0.05, respectively). However, they had no correlation with TIMP-1, TNF-α, OPG, or RANK. The MMP-9 serum level was found to be positively correlated with TNF-α level (r = 0.35, P < 0.05) and RANKL/OPG ratio (r = 0.27, P < 0.05) but not associated with RANKL. These results suggest that MMP-9, TNF-α, and the OPG/RANK/RANKL system may be closely interrelated and may play interactive roles in pathogenesis of osteoporosis in COPD.


Subject(s)
Matrix Metalloproteinase 9/metabolism , Osteoporosis/metabolism , Osteoprotegerin/metabolism , Pulmonary Disease, Chronic Obstructive/metabolism , RANK Ligand/metabolism , Receptor Activator of Nuclear Factor-kappa B/metabolism , Tumor Necrosis Factor-alpha/metabolism , Aged , Body Mass Index , Bone Density , Case-Control Studies , Humans , Male , Middle Aged , Osteoporosis/complications , Pulmonary Disease, Chronic Obstructive/complications , Tissue Inhibitor of Metalloproteinase-1/metabolism
10.
Zhonghua Yi Xue Za Zhi ; 92(12): 818-21, 2012 Mar 27.
Article in Chinese | MEDLINE | ID: mdl-22781454

ABSTRACT

OBJECTIVE: To explore the post-therapeutic change of cathelicidin LL-37 in asthmatics of different inflammatory phenotypes. METHODS: Thirty-four patients with initially diagnosed asthma (asthma group) and 14 normal subjects (control group) were recruited at Nanfang Hospital from August 2009 to August 2010 for this prospective study. Sputum and venous blood samples were collected and analyzed for cell differential. Eosinophilic asthma was defined as the count of sputum eosinophils ≥ 3%. The LL-37 concentrations in plasma and sputum supernatant were measured by enzyme-linked immunosorbent assay (ELISA) kit. The subjects were treated with budesonide/formoterol (160/4.5 µg) one inhalation twice daily and re-examined after 1 month. RESULTS: Prior to treatment, there were no differences between the asthma and control groups in the levels of LL-37 in plasma and sputum supernatant (P = 0.427,0.427). The plasma concentrations of LL-37 in asthma group were negatively correlated with baseline forced expiratory volume in one second (FEV(1), r = -0.470, P = 0.005), percent predicted of FEV(1) (FEV(1)%pred, r = -0.421, P = 0.013) and forced vital capacity (FVC, r = -0.367, P = 0.033). After treatment, the plasma and sputum supernatant concentrations of LL-37 (M (Q(R))) in the asthma group (5.6 (16.2), 65.6 (184.0) µg/L) were significantly higher than those baseline concentrations (5.03 (9.21), 28.40(109.76) µg/L, P = 0.005, 0.015). In the eosinophilic asthma subgroup, the plasma and sputum supernatant concentrations of LL-37 (M (Q(R))) after treatment (5.3 (19.3), 65.6 (185.2) µg/L) were significantly higher than those baseline concentrations (6.7 (8.9) L, 35.3 (102.0) µg/L, P = 0.021,0.014). And in the non-eosinophilic asthma subgroup, the changes of plasma and sputum supernatant concentrations of LL-37 showed no significant differences (P = 0.139, 0.386). In the asthma group, the correlations between plasma concentrations of LL-37 and FEV(1), FEV(1)%pred, FVC were not statistically significant (P = 0.283, 0.706,0.272) after treatment. CONCLUSIONS: LL-37 may participate in the aggravation of asthma. The elevated concentrations of LL-37 in eosinophilic asthma is probably due to the resolved suppression of LL-37 expression by eosinophilic inflammation. But its mechanism needs further researches.


Subject(s)
Antimicrobial Cationic Peptides/metabolism , Asthma/metabolism , Asthma/therapy , Adult , Asthma/pathology , Case-Control Studies , Female , Humans , Inflammation , Male , Middle Aged , Prospective Studies , Cathelicidins
11.
Nan Fang Yi Ke Da Xue Xue Bao ; 31(7): 1187-9, 2011 Jun.
Article in Chinese | MEDLINE | ID: mdl-21764691

ABSTRACT

OBJECTIVE: To observe the effect of 25-hydroxyvitamin D3 on the permeability and ZO-1 expression in normal human airway epithelial cells. METHODS: MTT assay was used to assess the viability of human airway epithelial cell line 16HBE following a 24-hour exposure to different concentrations of 25-hydroxy vitamin D3, and the transepithelial electrical resistance (TER) of the cell monolayer was measured using a Millicell-ERS voltohmmeter. Real-time quantitative RT-PCR was employed to determine the changes of ZO-1 mRNA expression in the cells following the exposures. RESULTS: Exposure to 25-hydroxyvitamin D3 resulted in significantly increased permeability of 16HBE cells, but the exspression of ZO-1 showed no obvious changes. 25-hydroxyvitamin D3 at 4×10(-9) mol/L showed the strongest effect in increasing the permeability of cell monolayer. CONCLUSION: 25-hydroxyvitamin D3 increases the permeability of normal bronchial airway epithelial cell monolayer in vitro, but this effect is not mediated by upregulation of ZO-1 expression.


Subject(s)
Bronchi/cytology , Calcifediol/pharmacology , Cell Membrane Permeability/drug effects , Epithelial Cells/metabolism , Zonula Occludens-1 Protein/metabolism , Bronchi/metabolism , Calcifediol/pharmacokinetics , Cell Line , Epithelial Cells/cytology , Humans , RNA, Messenger/genetics , RNA, Messenger/metabolism , Zonula Occludens-1 Protein/genetics
12.
Nan Fang Yi Ke Da Xue Xue Bao ; 31(6): 928-32, 2011 Jun.
Article in Chinese | MEDLINE | ID: mdl-21690039

ABSTRACT

OBJECTIVE: Eosinophils play a pivotal role in asthmatic airway inflammation. We previously found a significantly high expression of Slingshot-1L (SSH-1L) in peripheral eosinophils in acute exacerbations of asthma. Objective To investigate the expression and localization patterns of SSH-1L in peripheral blood eosinophils of asthmatic patients and their changes after treatment with inhaled corticosteroids. METHODS: We recruited 4 outpatients with acute exacerbations of asthma who received no previous corticosteroid treatment and 1 healthy volunteer. From all the subjects 30 ml peripheral venous blood samples were collected before and after a 3-month treatment with inhaled fluticasone. The eosinophils were isolated, purified and counted, and the expressions of SSH-1L in the eosinophils were examined by RT-PCR and Western blotting. The localization of SSH-1L phosphatases in the peripheral eosinophils was detected by immunofluorescence assay in one patient. RESULTS: SSH-1L phosphatases distributed diffusely in the cytoplasm, especially dense near the membrane of the peripheral eosinophils. Glucocorticoids treatment resulted in a significant reduction in both the SSH-1L mRNA expression (0.7403∓0.1124 vs 0.4101∓0.0363, P=0.001) and SSH-1L protein expression (0.3410∓0.1337 vs 0.1543∓0.0551, P=0.039). CONCLUSION: A high expression of SSH-1L in peripheral eosinophils in acute exacerbations of asthma may play a role in the activation and migration of eosinophils. The efficacy of inhaled corticosteroids in asthma control might be partly attributed to a down-regulated expression of SSH-1L.


Subject(s)
Asthma/blood , Eosinophils/metabolism , Phosphoprotein Phosphatases/metabolism , Adult , Aged , Asthma/drug therapy , Female , Glucocorticoids/therapeutic use , Humans , Male , Middle Aged
13.
Nan Fang Yi Ke Da Xue Xue Bao ; 31(4): 641-4, 2011 Apr.
Article in Chinese | MEDLINE | ID: mdl-21515460

ABSTRACT

OBJECTIVE: To investigate the level of the patients perceived control of asthma (PCA) in South China and analyze the risk factors contributing to inadequate PCA. METHODS: A total of 150 asthmatic out-patients consisting of 86 males and 64 females aged 19-65 (38.6∓11.7) years were enrolled in this investigation. The patients were asked to complete questionnaires of the demographic data, perceived control of asthma (PCAQ-6) scales, asthma control test (ACT) scales and Standard asthma-specific quality of life [AQLQ(S)] scale. The data of spirometric measurements, blood cell count and induced sputum cell count were also collected. RESULTS: All the 150 asthmatic out-patients recruited completed the questionnaires and examinations. The PCAQ-6 scores ranged from 10 to 26 (18.75∓3.42) in these patients (18.6∓3.28 in male and 18.95∓3.6 in female patients), significantly lower than those reported in other countries (P<1). PCA was positively correlated to the level of asthma control (r(p)=0.377, P=0.000) and AQLQ(S) scores (r(p)=0.675, P=0.000). Multiple linear regression showed that PCA was positively correlated to FEV1% and blood neutrophil counts, and inversely to asthma duration. CONCLUSION: The level of the PCA appears inadequate in South China. The PCA can affect the level of asthma control and asthma-specific quality of life. The factors contributing to inadequate PCA include primarily asthma duration, lung function and blood neutrophil counts.


Subject(s)
Asthma/prevention & control , Asthma/psychology , Health Knowledge, Attitudes, Practice , Surveys and Questionnaires , Adult , Aged , Asthma/blood , China , Female , Humans , Male , Middle Aged , Neutrophils , Quality of Life , Young Adult
14.
Nan Fang Yi Ke Da Xue Xue Bao ; 31(2): 239-43, 2011 Feb.
Article in Chinese | MEDLINE | ID: mdl-21354901

ABSTRACT

OBJECTIVE: To investigate the effect of toluene diisocyanate (TDI) on the production of reactive oxygen species (ROS) and the permeability of human bronchial epithelial (HBE) cells. METHODS: TDI-human serum albumin (TDI-HSA) conjugate was prepared using a modified Son's method. MTT assay was used to assess HBE cell viability after exposure to different concentrations of TDI-HSA. The level of intracellular ROS of HBE cells was detected by flow cytometry with an oxidation-sensitive fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) uploading, and the permeability of cell monolayer was assessed by detecting the transepithelial electrical resistance (TEER). RESULTS: The exposure to 120 µg/ml TDI-HSA did not obviously affect the cell viability. Compared with the control group, the intracellular fluorescent intensity increased significantly in the cells exposed to 20, 60, and 100 µg/ml TDI-HSA (P<0.05). The intracellular ROS production increased significantly after 100 µg/ml TDI-HSA treatment (P<0.05), but the increment in ROS production was significantly suppressed by pretreatment of the cells with N-acetylcysteine (NAC) (P<0.05), which also enhanced the TEER decreased by TDI-HSA treatment (P<0.05). CONCLUSIONS: TDI enhances the permeability of HBE cell monolayer partially through a ROS-mediated pathway, suggesting the importance of oxidative stress in TDI-induced pulmonary diseases.


Subject(s)
Bronchi/cytology , Cell Membrane Permeability/drug effects , Epithelial Cells/metabolism , Reactive Oxygen Species/metabolism , Toluene 2,4-Diisocyanate/pharmacology , Cell Line , Epithelial Cells/cytology , Humans , Oxidative Stress/drug effects , Serum Albumin/pharmacology
15.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(9): 2051-4, 2010 Sep.
Article in Chinese | MEDLINE | ID: mdl-20855247

ABSTRACT

OBJECTIVE: To investigate the expression of high mobility group box-1 (HMGB1) in the lung tissue and bronchoalveolar lavage fluid (BALF) of asthmatic mouse models and the influence of dexamethasone (DM). METHODS: Eighteen female Balb/C mice were randomly divided PBS control group, OVA group and OVA/DM group, and asthmatic mouse models were established in the latter two groups. The airway responsiveness of the mice was assessed by whole-body plethysmography, and the cells in the BALF were counted and classified, with the supernatants of the BALF collected for detection of the level of HMGB1 by ELISA. The left lung of the mice was collected for HE staining, and the expression of HMGB1 in the right lung tissue was detected by Western blotting. RESULTS: Asthmatic mouse models were successfully established. The level of HMGB1 in the BALF was significantly higher in OVA group than in the control group (6.31 ± 4.05 ng/ml vs 2.59 ± 0.73 ng/ml, P = 0.017), but no significant difference was found between OVA/DM group (3.39 ± 0.50 ng/ml) and OVA group (PP = 0.052). The expression of HMGB1 relative to tubulin was significantly higher in OVA group than in the control group (2.08 ± 0.87 vs 0.85 ± 0.30, P = 0.032), but similar between OVA/DM group (1.15 ± 0.48) and OVA group (PP = 0.133). CONCLUSION: The expression of HMGB1 is obviously increased in the lung and BALF of asthmatic mice and DM produces no significant effect on HMGB1 expression, suggesting that HMGB1 may serve as a new therapeutic target for asthma treatment.


Subject(s)
Asthma/metabolism , Dexamethasone/therapeutic use , HMGB1 Protein/metabolism , Lung/metabolism , Animals , Asthma/drug therapy , Bronchoalveolar Lavage Fluid/chemistry , Female , HMGB1 Protein/genetics , Mice , Mice, Inbred BALB C
16.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(9): 2084-6, 2010 Sep.
Article in Chinese | MEDLINE | ID: mdl-20855256

ABSTRACT

OBJECTIVE: To investigate the clinical indications of asthma control test (ACT). METHODS: A total of 120 asthmatic patients with a diagnosis in line with the American Thoracic Society criteria and treated for over a month were enrolled in this study. The patients were asked to complete a survey to assess their symptoms and asthma attacks, and ACT evaluation was conducted by physicians familiar with ACT evaluation. The patients were classified into two groups based on the pulmonary function test (positive for bronchodilator test and provocation test) or based on disease severity (mild and moderate-to-severe asthma groups). The effect of ACT evaluation was graded as good (no less than 4 item available for evaluation), fair (2-3 items available) and poor (no more than 1 item). To further analyze the ACT sensitivity in relation to different disease severity, 29 asthmatic patients with an initial diagnosis and BDT positivity were included, and the ACT score of the patients with mild, moderate and severe asthma based on FEV1% were compared. RESULTS: In patients positive for bronchodilator test, good, fair and poor evaluation effects were found in 48, 15, and 5 cases, as compared to 10, 15, and 27 in those positive for provocation test, respectively, showing significant differences between the two groups (P < 0.001). In mild asthma group, good, fair and poor evaluation effects were found in 12, 15, and 18 cases, respectively, significantly different from those in moderate-to- severe asthma group (50, 21, and 4 cases, P < 0.001). ACT scores showed a positive correlation to FEV1% in 29 patients with positive BDT (r = 0.55, P = 0.003). ACT scores had no significant difference between mild and moderate asthma groups (P > 0.05), but showed significant differences between mild and severe groups (P = 0.009) and between moderate and severe groups (P = 0.008). CONCLUSION: ACT is more suitable for evaluating patients positive for bronchodilator test or with moderate to severe asthma.


Subject(s)
Asthma/diagnosis , Asthma/physiopathology , Severity of Illness Index , Surveys and Questionnaires , Adolescent , Adult , Aged , Female , Humans , Male , Mass Screening , Middle Aged , Predictive Value of Tests , Respiratory Function Tests , Sensitivity and Specificity , Young Adult
17.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(5): 963-5, 2010 May.
Article in Chinese | MEDLINE | ID: mdl-20501368

ABSTRACT

OBJECTIVE: To investigate the effect of dimethylsulfoxide on the expression of thymic stromal lymphopoietin (TSLP) in human bronchial epithelial cell (HBE). METHODS: 16HBE cells were incubated in the presence of dimethylsulfoxide at different concentrations, and the cell proliferation changes were observed. The expressions of TSLP mRNA and protein in the cells were detected by real-time quantitative PCR and ELISA, respectively. RESULTS: Dimethylsulfoxide induced significantly increased TSLP mRNA expression in HBE cells (P<0.01) in a concentration-dependent manner. The level of TSLP protein in the supernatant was also increased after dimethylsulfoxide treatment, but high concentration of dimethylsulfoxide resulted in e inhibited cell proliferation. CONCLUSION: Dimethylsulfoxide may affect the immunomodulatory function of HBE cells.


Subject(s)
Bronchi/metabolism , Cytokines/metabolism , Dimethyl Sulfoxide/pharmacology , Epithelial Cells/metabolism , Bronchi/cytology , Cell Proliferation/drug effects , Cells, Cultured , Cytokines/genetics , Humans , RNA, Messenger/genetics , RNA, Messenger/metabolism , Thymic Stromal Lymphopoietin
18.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(5): 1072-4, 2010 May.
Article in Chinese | MEDLINE | ID: mdl-20501397

ABSTRACT

OBJECTIVE: To observe the therapeutic effect of tiotropium bromide powder inhalation on stable bronchiectasis. METHODS: Twenty-two patients with stable bronchiectasis received inhalation of totropium bromide powder at the daily dose of 18 microg, and on days 1 and 28, the patients were examined for forced expiratory volume in one second (FEVl), predicted value [FEVl(%)], forced expiratory volume (FEV), and FEVl/FVC. The symptom score and BODE index were also recorded. RESULTS: After 1 month of inhalation therapy, the FEV1% of the patients showed a moderate increase but the increment was not statistically significant (t=-1.875, P>0.05); the symptom score and BODE index decreased significantly after the therapy (t=7.091, P<0.001; t=2.982, P<0.05). CONCLUSION: Long-term inhalation of tiotropium bromide powder can improve the clinical symptoms and BODE index and enhance the exercise tolerance and quality of life of the patients with bronchiectasis.


Subject(s)
Bronchiectasis/drug therapy , Receptor, Muscarinic M3/antagonists & inhibitors , Scopolamine Derivatives/administration & dosage , Administration, Inhalation , Adult , Aged , Female , Forced Expiratory Volume , Humans , Male , Middle Aged , Powders , Tiotropium Bromide
19.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(4): 700-3, 2010 Apr.
Article in Chinese | MEDLINE | ID: mdl-20423829

ABSTRACT

OBJECTIVE: To investigate the expression and release of high mobility group Box-1 protein (HMGB1) in the lung tissue of mice with respiratory syncytial virus (RSV) infection. METHODS: Eighteen mice were randomized into PBS control group, RSV group and RSV/ribavirin group. Seven days after RSV infection in the mice in the latter two groups, the bronchoalveolar lavage fluid (BALF) was collected for cell counting and classification, and the levels of IL-4, IFN-gamma and HMGB1 in the supernatants of the BALF were detected. The left lungs of the mice were harvested for pathological examination with HE staining, and the right lungs were taken for detecting the expression of HMGB1 by Western blotting. RESULTS: RSV induced a TH1 inflammation in the lung tissue as shown by significantly increased IFN-gamma and decreased IL-4 levels in the BALF. The total BALF cells, neutrophils and macrophages in the RSV group were significantly higher than those in the control group (P<0.05), and the cell counts were significantly decreased by ribavirin treatment (P<0.05). HE staining showed neutrophil and lymphocyte infiltration in the lumen and submucous layer of the airway in RSV group. The level of HMGB1 in the BALF significantly increased in the RSV group as compared with that in the control group (P<0.05), but was lowered by ribavirin treatment (P<0.05). The expression of the HMGB1 in the lung tissue significantly increased in the RSV group in comparison with that in the control group (P<0.05), and was not significantly decreased by ribavirin treatment (P>0.05). CONCLUSIONS: The increased expression and release of HMGB1 in the lung tissue of mice with RSV infection is probably involved in the development of RSV infection-related lung diseases.


Subject(s)
HMGB1 Protein/biosynthesis , Lung/metabolism , Respiratory Syncytial Virus Infections/metabolism , Animals , Bronchoalveolar Lavage Fluid/chemistry , Female , HMGB1 Protein/genetics , Mice , Mice, Inbred BALB C , Random Allocation
20.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(2): 228-31, 2010 Feb.
Article in Chinese | MEDLINE | ID: mdl-20159686

ABSTRACT

OBJECTIVE: To investigate the effect of hydrogen dioxide (H(2)O(2)) on the expression of vascular endothelial growth factor (VEGF) in human bronchiolar epithelial (HBE) cells. METHODS: MTT assay was used to assess HBE cell viability after exposure to different concentrations of H(2)O(2). VEGF/beta-actin gene fragments were amplified simultaneously by RT-PCR from the total HBE cell RNA, and VEGF protein expression in the cells was detected using ELISA. RESULTS: The exposure to 200 micromol/L H(2)O(2) did not obviously affected the cell viability. Compared with those in the control cell, VEGF165/beta-actin and VEGF189/beta-actin ratios were significantly increased in the cells after treatment with 50, 200, and 600 micromol/L H(2)O(2) (P<0.05). The protein expression of VEGF significantly increased after 50 micromol/L H(2)O(2) treatment (P<0.05), but significantly decreased with pretreatment with the PI3K inhibitor Ly294002 (P>0.05). CONCLUSION: Oxidative stress increases the expression of VEGF via a PI3K-dependent pathway in human bronchiolar epithelial cells, which may play an important role in the onset and maintenance of chronic inflammation in asthma.


Subject(s)
Bronchi/cytology , Epithelial Cells/metabolism , Oxidative Stress/physiology , Phosphatidylinositol 3-Kinases/metabolism , Vascular Endothelial Growth Factor A/metabolism , Actins/metabolism , Cell Line , Epithelial Cells/cytology , Humans , Hydrogen Peroxide/pharmacology , Signal Transduction , Vascular Endothelial Growth Factor A/genetics
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