ObjectiveTo investigate the effects of nicotine on the expression of Th1/Th2 cytokines and transcription factor T-bet/GATA-3 in cultured CD4+ T of rat sensitized by ovalbumin.Methods Two weeks after immunization by ovalbumin,splenic CD4+ T cells of Wistar rat were purified using CD4+T cell enrichment kit.Purified CD4+ T cells of rat were cultured and divided into 4 groups:a control group,1 μg/ml nicotine stimulated group,10 μg/ml nicotine stimulated group,100 μg/ml nicotine stimulated group.These cells,in their groups,were stimulated with or without nicotine and were all challenged simultaneously with OVA.Supernatants and cell pellets were harvested after being stimulated for 24 h.The concentration of IFN-γ and IL-4 in supernatants were measured by enzyme-linked immunosorbent assay (ELISA).Real-time PCR was used to determine the mRNA expression of T-bet and GATA-3 in CD4+T cells.Results ( 1 ) IFN-γproduction was significantly decreased in all nicotine treated groups [ ( 113.78±6.06) ng/L,(70.31±7.26) ng/L,(20.00±2.14) ng/L] compared with the control group[ (142.30± 5.89) ng/L],and the level of IL-4 was significantly increased in all nicotine treated groups [ (69.49±3.91) ng/L,(93.63±4.56) ng/L,(50.97±3.07) ng/L] compared with the control group[ (36.91±3.24) ng/L].(2) Expression of T-bet mRNA in all nicotine treated groups(0.73±0.03,0.57±0.04,0.31 ±0.00) was lower than that in the control group(0.98±0.09),but expression of GATA-3 mRNA in all nicotine treated groups (4.31±0.26,5.16±0.23,1.56±0.14) was significantly higher than that in the control group(1.00±0.07).Conclusion Nicotine may play a key role in the development of Th2-type allergic inflammation in asthma by promoting over-expression of GATA-3 mRNA and downregulating the expression of T-bet mRNA.

ObjectiveTo explore the influence of the cigarette smoke in the airway inflammation and pulmonary function of the asthmatic patients. MethodsTwenty-five cases of asthmatic patients with cigarette smoke exposure, 22 cases of asthmatic patients without cigarette smoke exposure and 20 cases of normal control persons were involved in this study. The proportion of various inflammatory cells in the induced sputum, the levels of serum interleukin (IL)-8 and IL-4 and lung function (FEV1％ expected value,FEV1/ＦVC％ ) were detected. ResultsThe infiltrating of neutrophils was primarily found in sputum of the asthmatic patients with cigarette smoke exposure, but the infiltrating of eosinophils was mainly in sputum of the asthmatic patients without cigarette smoke exposure. The levels of serum IL-8 and IL-4 of peripheral blood of asthmatic patients with cigarette smoke exposure[(277.02 ±71.37), (171.69 ±31.01) ng/L] were significantly higher than those in asthmatic patients without cigarette smoke exposure[(158.88 ± 21.95 ),( 111.42 ± 21.69 ) ng/L] and normal control persons [( 116.78 ± 71.37 ), (73.94 ± 15.72 ) ng/L] (P ＜ 0.01 ).The FEV1％ expected value and FEV1/FVC％ of the asthmatic patients with cigarette smoke exposure [(51.12 ± 13.30) ％, ( 49.16 ± 11.09 )％] was lower than those of asthmatic patients without cigarette smokeexposure [(81.81 ± 5.82)％, (79.00 ± 3.86)％] and normal control persona [(95.50 ± 10.11 )％, (83.18 ±6.04)％] (P ＜ 0.01 ). The level of serum IL-8 was positively correlated to the neutrophils percentage in the induced sputum (r =0.742,P＜ 0.01 ) ,while negatively correlated to the FEV1％ expected value(r =-0.739,P ＜ 0.01 ). ConclusionCigarette smoke may influence the airway inflammation of the asthmatic patients and accelerate the deterioration of their lung function by promoting the producing of IL-8.

Objective To evaluate the effect of the activity of Ca2+/CaN-NFATc on the activation and prolfferation of lymphocyte in asthmatic rats.Methods The rats of the asthma group and the CsA group were sensitized and challenged by ovalbumin.So did the control group with saline instead.Lymphocyte was separated from spleen and cultured for 24 hours,and PHA-p (5 μg/ml) was added to the culture medium in every group,CsA ( 1.0 μg/ml) was added to the CsA group,respectively.The concentration of [ Ca2+ ] i,the activity of CaN,the protein expression of dephosphorylated NFATc and Cyclin E in T lymphocyte were assayed.The level of IL-4 and IL-2 in culture supernatants was measured,and the cell cycle distribution of lymphocyte was analyzed.ResultsWhen compared to the control group,the activity of Ca2+/CaN-NFATc [ (81.21 4-14.39) vs (63.66 ±9.02) ] was increased and the protein expression of CyclinE [ (0.9327 ±0.0370) vs (0.8374 ±0.0637) ] was higher in Lymphocyte of the asthma group ( P＜0.05,P ＜0.01,respectively).The percentage of lymphocyte in the S phase [ (7.8600±2.8241) vs (4.0270 ± 1.8650) ] and S + G2/M phase [ ( 10.6700±3.3850) vs (5.8740 ± 1.4389) ] was higher;however,the percentage of G0/G1 phase [ (89.3300 ± 3.3850) vs (94.1260± 1.4389 )] was lower in asthma group ( all P ＜ 0.01 ).The level of IL-4 [ ( 1.55 ± 0.19) pg/ml vs (0.99 ± 0.12 ) pg/ml ] and the IL-4/ IL-2 ratio [ (0.81 ±0.12) vs (0.49 ±0.49) ] in culture supernatants of the asthma group were higher than those of the control group ( all P ＜0.01 ).While the activity of Ca2+/CaN-NFATc (47.19 ±7.16)was decreased and the protein expression of Cyclin E (0.6840 ± 0.0485 ) was reduced in lymphocyte in CsA group(all P ＜0.01 ),the percentage of lymphocyte in the S phase (4.8600 ± 1.9595) and S + G2/M phase (7.9900 ± 1.9405) was decreased and the percentage of G0/G1 phase (92.2100 ± 1.9267) was increased ( all P ＜ 0.05 ),and the level of IL-4 (0.47 ± 0.09 ) pg/ml and the ratio of IL-4/ IL-2 (0.78 ±0.20) was lower in culture supernatants in the CsA group than that of the asthma group( all P ＜ 0.01 ).There was a positive correlation between the protein expression of dephosphorylated NFATc and the protein expression of CyclinE in Lymphocyte,so did between the protein expression of NFATc and the level of IL-4 in culture supernatants( r =0.711,P ＜0.01 and r =0.749,P ＜0.01.respectively).Conclusions The activity of Ca2+/CaN-NFATc was increased in lymphocyte of the asthmatic rats.Its increasing might result in the imbalance of Th1/Th2 by promoting the expression of IL-4 and might lead to the proliferation of lymphocyte by promoting the Cyclin E expression.

Objective To evaluate the activity of Ca2+ /CaN-NFATc, and study its association with the imbalance of TH1/ TH2 in asthmatic rat lungs. Methods Twenty-four Wistar rats were random divided to the asthma group and control group, twelve rats each group. The rats were sensitized and challenged with ovalbumin to establish the asthmatic model. The pulmonary function of rats was surveyed and evaluated u-sing Maclab system. Airway inflammation and the thickness of bronchial wall ( WAt/Pi) were observed by H. E staining. The quantity of Ca2+ , the activity of CaN , the protein expression of dephosphorylated NFATc and the level of IL-4 and IL-2 were assayed.Results Compared with control group, the thickness of bronchial wall was significantly increased ( t = -7. 99, P <0. 01), the airway resistance was higher( t = 2.59, P <0.05) and the respiration frequency was faster( t =7.94, P <0.01) ,but the minute ventilation volume was lower( t =6. 87, P <0.01) in asthma group. The levels of IL-4 and the IL-4/ IL-2 ratio in rat lungs of asthma group were significantly higher than those in control group ( t = -8.69, P <0. 01; t = 11.40, P <0. 01 .respectively) , however, the levels of IL-2 in asthma group were lower than that in control group ( t =8. 29, P <0. 01). The activity of CaN and the protein expression of dephosphorylated NFATc in asthma group were higher than those in control group( t = -2. 91, P <0.01; t = -22.45, P <0.01,respectively) ,but the quantity of Ca2+ in asthma group was lower than that in control group( t =4. 747, P < 0.01). There wag a positive correlation between the activity of CaN and the protein expression of dephos-phorylated NFATc( r =0. 39, P <0.05) ,so did between the protein expression of dephosphorylated NFATc and the IL-4/ IL-2 ratio( r =0. 83-, P <0.01) ,and the same between the thickness of bronchial wall and the IL-4/ IL-2 ratio( r = 0. 84, P < 0.01). Conclusions The activity of CaN-NFATc was increased in rat lungs of asthma group, and the rising of which might increase the ratio IL-4/ IL-2. Thus, the signal of CaN-NFATc probably took part in the imbalance of TH1/ TH2 in asthmatic rat lungs.

Objective To investigate the impact of cigarette smoking on inflammatary factors in sputum including cell component,IL-8 and eotaxin,and the effect on responses to treatment with inhaled corticosteroids (ICS)in patients with asthma.Methods Thirty-eight outpatients with chronic stable asthma who visited from January 2009 to February 2010 were enrolled in the study.Twenty-three cases were nonsmokers and 15 cases were smokers.All of them were treated by daily inhaled budesonide,and[32 agonist when necessary.Induced sputum eosinophil and neutrophil proportion were measured,and the levels of interleukin(IL-8)and eotaxin in induced-sputum by enzymatic immunoassay(ELISA)were compared between non-smoking and smoking asthmatic patients.Results The eosinophil proportion was(2.0 ± 0.4)％ in smokers and(4.6 ± 2.5)％ in nonsmokers before therapy,and(1.1 ± 0.5)％ in smokers and(1.8 ± 0.6)％ in nonsmokers after therapy.The difference was statistical significant(F =15.271,P ＜ 0.05).The eotaxin was(3.5 ± 2.1)× 10-3 mg/L insmokers and(8.6 + 2.3)x 10-3 mg/L in nonsmokers before therapy,and(3.1 + 1.5)x 10-3 mg/L insmokers)and(3.6 ± 1.3)x 10-3 mg/L in nonsmokers after therapy.There was statistical significant(F =24.172,P ＜ 0.05).There were no significant difference on neutrophil proportion and IL-8 before and after treatment(F =1.563 and 1.793,respectively,Ps ＞ 0.05).Neutrophil proportion(F =9.632,P ＜ 0.05)and IL-8(F =5.720,P ＜ 0.05)in smokers were significantly higher than non-smokers,whereas eosinophil proportion (F =15.879,P ＜ 0.05)and eotaxin(F =12.365,P ＜ 0.05)in smokers were significantly lower than nonsmokers.Conclusion There was a significantly increase in inflammatary cells andfactors in induced-sputum in two groups after ICS treatment,but the effect in smokers was lower than in non-smokers.Neutrophil proportion and IL-8 were higher in induced sputum in smokers,whereas eosinophil proportion and eotaxin were higher in non-smokers.

AIM:To observe the expression of cyclin E in the lungs of asthmatic rats and to evaluate the role of cyclin E in airway remodeling with asthma. METHODS:Twenty Wistar rats were randomized to the asthma group and control group (10 rats in each group). The rats were sensitized and challenged with ovalbumin to establish the asthmatic model. Airway inflammation was observed by HE staining. The thickness of the bronchial wall (WA/Pi) was measured by computer-assisted image analysis system. The cell cycle distribution of monocytes in peripheral blood was analyzed by flow cytometry. The mRNA expression of cyclin E was detected by realtime RT-PCR. The protein expression of cyclin E was assayed by Western blotting and immunohistochemistry. RESULTS:The thickness of the bronchial wall in asthma group was significantly higher than that in control group (P

AIM: To investigate the expression of interleukin-13 (IL-13) in lung tissue of rat asthmatic model and effect of triptolide on expression of IL-13. METHODS: A asthmatic model was established and inflammation changes of lung tissues were examined by haematoxylin-eosin staining. Reverse transcriptio polymerase chain reaction (RT-PCR) was used to semiquantitate the expression of IL-13 mRNA, and enzyme-linked-immuno-sorbent assay (ELISA) was used to detect protein level of IL-13. RESULTS: There was a significant increase in the count of EOS, the expression of IL-13 mRNA and IL-13 protein level in lung tissue of asthmatic rats compared with those of triptolide-treated rats (P0.05). The expression of IL-13 mRNA and IL-13 protein level in triptolide-treated rats were significant lower than that of asthmatic rats (P

Objective To observe the content of macrophage inflammatory protein-1α(MIP-1α)in plasma and bronchoalveolar lavage fluid (BALF) and the expressions of MIP-1α mRNA and nuclear factor-kappa B( NFκB)p65 mRNA in the lung of rats subjected to mechanical ventilation with high tidal volume. Method Thirty-two healthy Wistar rats were randomly ( random number) divided into control group, ventilator induced lung injury (VILI) group,dexamethasone (DEX) group and budesonide (BUD) group. The content of MIP-1α in plasma and BALF were measured with ELJSA and the expressions of MIP-1α mRNA and NF-κBp65 mRNA in lung of rat were detected by RT-PCR. The data distributed were expressed as (-x) ± s and were compared among 4 groups. Furthermore, the correlation between the content of MIP-1α and the expression of MIP-1α mRNA, and the correlation between the expression of MIP-1α mRNA and the expression of NF-κBp65 mRNA were analyzed in the latter three groups. Results With lessened lung injury ,the content of MIP-1αin plasma and BALF and the expressions of MIP-1α mRNA and NF-κBp65 mRNA in lungs of rats of DEX and BUD groups were significantly lower than those in VILI group ( P ＜ 0.001 ). Although the content of MIP-1α in plasma and BALF and the expressions of MIP-1α mRNA and NF-κBp65 mRNA in lungs of rats of BUD group were higher than those of DEX group, but no significant differences were found between them ( P ＞ 0.05). Correlation study showed that positive correlations were xisted between the MIP-1α in plasma and the expression of MIP-1α mRNA in the lungs ( r = 0.895, P ＜ 0.05)and between the expression of MIP-1α mRNA and the expression of NF-κBp65 mRNA in the lungs ( r=0.801, P ＜ 0.05). Conclusions Glucocorticoid could down-regulate the expression of MIP-1α mRNA by inhibiting the activity of NF-κB in the lungs and may have preventive and therapeutic effect to VILI to some extent. The effect of glucocorticoid used locally against VILI is simnilar to that of systemic administration with lesser adverse reactions.

Objective To investigate the feasibility and effects of endoscopic surgical treatment of lumbar intervertebral disc herniation associated with veitebral osteochondrosis.Methods From June 2008 to December 2015,276 cases of lumbar intervertebral disc herniation associated with vertebral osteochondrosis were treated with endoscopic surgery,including 185 men and 91 women,with an average 39.2 years old (range,16-65 years old).The involved level included L2.3 in 2 cases,L3.4 in 9 cases,L4,5 in 126 cases and L5S1 in 139 cases.On preoperative axial CT,the diameter of ossification was more than half of the transverse or sagittal diameter of the spinal canal in 89 cases,and no more than half of the transverse and sagittal diameter of the spinal canal in 187 cases.All patients were operated on the side with serious symptom,181 cases were operated with mobile microendoscopic discectomy (MMED),and 95 cases were operated with percutaneous endoscopic surgery,including percutaneous transforaminal endoscopic discectomy (PTED) in 61 cases and the percutaneous interlaminar endoscopic discectomy (PIED) in 34 cases.The operation and complications were analyzed.Results The soft herniation,broken disc material and the periphery of compressing ossification were removed under the endoscope in all cases,until the nerve was well decompressed.However,the ossification was not complete resected.Dural sac tear occurred in 3 cases of MMED.In the early stage of PTED,2 cases converted to MMED because of intraoperative pain and difficulty,and one case had exiting nerve root injury.At the final follow-up of 12-60 months (average,20.6 months),visual analogue scale decreased from preoperative 8.5±1.2 to 1.0±0.9,Oswestry disability index decreased from preoperative 40.2±8.6 to 3.1±3.0.According to Macnab scale,the results were excellent in 89,good in 154 cases,moderate in 33 cases.Conclusion For most lumbar intervertebral disc herniation associated with vertebral osteochondrosis,good results can be achieve by removal of herniated and broken intervertebral disc and decompression of nerve with endoscope.Therefore,we speculate that the soft disc herniation and spinal stenosis are main pathogenic factors,and that the complete resection of ossification is not needed.

This study was aimed to optimize the ethanol extraction technology ofXiao-Xu-Ming (XXM) decoction. The extraction rates of paeoniflorin, ferulic acid, tetrandrine and yield extract were used as comprehensive evaluation indexes. The amount of ethanol, extraction times, extraction time and concentration of ethanol were selected as factors. The orthogonal design was used to optimize ethanol reflux extraction technology of XXM decoction. The results showed that the optimum extraction conditions were as follows: refluxing extracted 3 times with 10 folds, 70% ethanol, 1.5 h for each time. It was concluded that the optimized extraction process was objective, practical, reasonable and stable, which provided experimental basis for industrial production of XXM decoction.