To investigate the expression of mRNA and protein of telomerase reverse transcriptase(TERT) in guinea pig asthmatic lung tissues and human circulating lymphocytes and to discuss the role of TERT in the pathogenesis of asthma and the clinical significance of expression of TERT in human asthmatic circulating lymphocytes in monitoring the state of asthma.Methods: In 6 guinea pig asthmatic lung tissues, 6 guinea pig normal lung tissues, 16 samples of circulating lymphocytes of asthmatic patients and 14 normal circulating lymphocytes the expression of mRNA and protein of TERT were detected with nuclei acid in situ hybridization and immunostaining. The expression of TERT in circulating lymphocyte were compared with pulmonary function and serum IgE level. Results:The expression of mRNA and protein were up-regulated in asthmatic airway epithelium and lymph nodule germinal center in guinea pig. Compared with control group,high expression were also showed in human asthmatic circulating lymphocytes.The index of TERT expressive positive lymphocytes showed negative correlation with FEV1/ predictive value and MW/predicn've value,positive correlation with serum IgE level. Conclusion: (1 )TERT expression in asthmatic airway epithelium and lymph nodule germinal centers of guinea pig and human circulating lymphocytes were up-regulated; (2)Detection of TERT expression of asthmatic circulating lymphocytes may be useful to monitor the state of asthma.

The changes of CD4(+)CD25(+) regulatory T cells (CD4(+)CD25(+) Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4(+)CD25(+) Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4(+)CD25(+) Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4(+)CD25(+) Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control group (P<0.05). Although the CD4(+)CD25(+) Treg ratio and Foxp3 mRNA of remission group also lower than that of normal control group, there was no significant difference between them (P>0.05). As compared with persistent group, exacerbation group had lower CD4(+)CD25(+) Treg ratio and Foxp3 mRNA (P<0.05). It was indicated that the decrease of CD4(+)CD25(+) Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

Objective To investigate changes of CD~+_4CD~+_ 25 regulatory T cells (CD~+_4CD~+_ 25 Treg) and forkhead/winged helix transcription factor(Foxp3) mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma, so as to elucidate the possible roles of CD~+_4CD~+_ 25 Treg in the development of asthma. Methods The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD~+_4CD~+_ 25 Treg ratio and Foxp3 mRNA in PBMCs were detected. Results The CD~+_4CD~+_ 25 Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent group were lower than that of remission group and normal control group (P0.05). Compared with persistent group, exacerbation group had lower CD~+_4CD~+_ 25 Treg ratio and Foxp3 mRNA (P

AIM: To explore the change of delayed rectifier potassium channel (K_V) activity in alveolar macrophages (AM) in chronic obstructive pulmonary disease (COPD) rats. METHODS: COPD model was established by exposure of the animals to cigarette smoke. With whole-cell voltage- or current-clamp techniques, K_V activity, membrane capacitance and resting membrane potential (Em) in AM from COPD model and control rats were compared. RESULTS: (1) Significant increases in total mononuclear cells and AM in bronchoal aveolar lavage fluid (BALF) were found in COPD group compared with in control group. (2) The AM K_V current altitude in COPD group [(520.5?38.7)pA, (+50) mV, n=30] was significantly lower than that in control group [(713.6?44.4)pA, (+50) mV, n=30, P0.05), but had more positive Em (P

To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-kappa B were labelled with DIG by terminal transferase. After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8% nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged. Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film. The results showed that nuclear proteins binded specifically to the NF-kappa B consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-kappa B in PMA group was more than that in PMA + PDTC group. It is suggested that detection of NF-kappa B by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories.
Chemiluminescent Measurements ; DNA-Binding Proteins/*analysis ; DNA-Binding Proteins/genetics ; Electrophoretic Mobility Shift Assay ; NF-kappa B/*analysis ; NF-kappa B/genetics ; NF-kappa B/metabolism ; Rats, Sprague-Dawley ; *Trans-Activation (Genetics) ; Trans-Activators/analysis ; Trans-Activators/genetics ; *Transcription, Genetic

AIM:To observe the effects of prostaglandin E2 on transforming growth factor-?1(TGF-?1)triggered human lung fetus fibroblast(HLF)transdifferentiation and connective tissue growth factor(CTGF),collagen type I(COLⅠ)expression.METHODS:HLFs were treated with TGF-?1,the cells underwent phenotypic change to myofibroblast.The marker of myofibroblast-?-smooth muscle actin(?-SMA)was detected by immunofluorescence.The ?-SMA content was measured by Western blotting.The changes in CTGF and COL Ⅰ at transcription levels were estimated by RT-PCR method.CTGF protein expression was evaluated by immunocytochemical.Cell culture medium hydroxyproline amount was measured by colormetric assay.RESULTS:PGE2 blocked TGF-?1 induced ?-SMA positive myofibroblast transformation(P

To compare the efficacy, safety, and tolerability of intravenous moxifloxacin with those of a commonly used empirical antibiotic regimen, cefoperazone and azithromycin in the treatment of community acquired pneumonia (CAP) in adult patients requiring initial parenteral therapy, 40 patients with CAP were divided into two groups, a moxifloxacin group (n = 20) and a control group (n = 20), which were treated for 7 to 14 days. The patients in the moxifloxacin group were intravenously given 400 mg of moxifloxacin (Avelox) once a day. Patients in the control group were administered 2.0 g of cefoperazone twice a day and azithromycin 0.5 g once a day. Clinical, bacteriological, and laboratory examinations were performed before the treatment, and at the end of the treatment. Our results showed that there was no significant difference in the clinical efficacy rate between two treatment groups at end of therapy (90% for moxifloxacin, 95% for cefoperazone plus azithromycin) (P > 0.05). The bacteriologic eradication rate at the end of treatment was 90% in the moxifloxacin group and 80% in the cefoperazone-plus-azithromycin group, whereas there was no significant difference between the two groups (P > 0.05). In addition, both drugs were well-tolerated in this trial, with the number of drug-related adverse events being comparable. It is concluded that moxifloxacin is an effective and well-tolerated treatment for CAP and was equivalent to the commonly used empirical treatment of cefoperazone plus azithromycin. Moxifloxacin is likely to offer clinicians an alternative for reliable empirical CAP treatment in the face of increasing antibiotic resistance.

The transcription activity of ectogenic human carcinoembryonic antigen (CEA) promoter in lung adenocarcinoma cells A549 was investigated for the further gene-targeting therapy. The reporter gene green fluorescent protein (GFP) driven by CEA promoter and human cytomegalovirus (CMV) promoter were relatively constructed and named plasmid pCEA-EGFP and pCMV-GFP respectively. The intensity of fluorescence was detected by fluorescence microscope and flow cytometry analysis after the pCEA-GFP and pSNAV-GFP plasmids were transfected into A549 cells through liposome respectively. The results showed (4.08+/-0.63) % of the A549 cells transfected with pCEA-AFP plasmid expressed, significantly lower than that of the A549 cells transfected with pCMV-GFP [(43.27+/-3.54) %]. It was suggested that ectogenic human CEA promoter in lung adenocarcinoma cells A549 was weakly expressed. The distinct specificity of CEA promoter in CEA high expression cells was regarded as a tool in selective gene therapy, but the transcription activity of ectogenic human CEA promoter was needed to increase in the future.

In order to explore the expression of PI-3K in T lymphocytes of asthmatic rats and the relationship between PI-3K and activation of T lymphocytes, 24 Wistar rats were randomly divided into 4 groups: normal control group, asthmatic one-week group, asthmatic two-week group and asthmatic four-week group. T cells were purified from blood of each rat and the expression of PI-3K was observed by immunocytochemical fluorescence staining, the semiquantitative fluorescence intensity was measured by HPIAS-2000 analytic software, and the expression of IL-4 in supernatants was detected by ELISA. The results showed that the fluorescence intensity of T lymphocytes in asthmatic groups was significantly higher than that in normal control (P<0.001), indicating that the expression of PI-3K in T lymphocytes of asthmatic rats was significantly higher than that in those of normal controls, and the difference between acute and chronic stage asthmatic groups was significant (P<0.05). The expression levels of IL-4 protein in supernatants of asthmatic T lymphocytes were significantly higher than those in the normal controls (P<0.05). There was a significant positive correlation between the expression of PI-3K in T lymphocytes and the IL-4 protein expression in supernatants (r=0.583, P<0.01). It was suggested that PI-3K signal pathway may participate in the processes of activation and other cytological effects of asthmatic T lymphocytes, thus may play an important roles in the pathogenesis of asthma.

Objective To investigate the expression of CD40 and CD40L on the surface of peripheral blood mononuclear cells(PBMCs)in asthmatic rats and the effect of anti-CD40L McAb on cytokines of it.Methods Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs ih asthmatic rats.After the PBMCs Was treated with anti.CIMOL McAb.ELISA was used to detect the levels of IL-4 and IFN-γin the supematants of cultured cells.Results Compared with the normal control group.the expression of CD40 and CD40L of PBMCs in asthImatic rats increased(P＜0.05).Compared with the untreated group,the level of IL-4 and the ratio of IL4/IFN-γ decreased after the PBMCs were treated with anti-CD40L McAb(P＜0.05).Conclusion The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats Was unregulated.Anti-CD40L Mcab Can decrease the level of IL-4 and the ratio of IL_4/IFN-γ.