Objective To investigate the application value of tuberculosis infection of T cells spot test in the diagnosis for pulmonary tuberculosis in elderly patients with chronic obstructive pulmonary disease(COPD).Methods According to test different ways,64 elderly COPD patients with pulmonary tuberculosis were divided into the two groups.The control group(n =32)was checked by tuberculosis antibody test,and the study group(n =32)received T cells infected with tuberculosis spot test.The two sets of results were observed and compared.Results The study group by T cell spot test TB infection,29 cases were detected positive(90.63%),3 cases of negative(9.37%);tuberculosis antibody detection by the control group,23 cases were detected positive(71.88%),9 negative patients (28.12%).Compared the two groups,the study group had higher positive rate(χ2 =3.954 7,P <0.05),and nega-tive rate was lower(χ2 =3.914 7,P <0.05).Conclusion Tuberculosis infection T cell spot test in the diagnosis of pulmonary tuberculosis in patients with COPD in the elderly has greater value,more accurate diagnosis,reliability and worthy of promotion.

Objective To observe clinical efficacy of Changyunning granula in treatment ofulcerative colitis.Methods 90 cases from March 2014 to August 2015 were randomly divided into observation group and control group of 45 patients in control group received conventional Western medicine treatment, observation group received more Granule intestinal treatment, patients were followed up and recorded Ig and C3 change the situation serum TNF-αand IL-8 changes, colonoscopy curative effect.Results The study group patients IgA, IgG, IgM, C3, respectively (3.65 ±0.48,13.41 ±2.36, 1.98 ±0.39,1.15 ±0.32)g/L, were better than control group (3.78 ±0.39,15.69 ±2.41, 2.45 ±0.36,1.63 ±0.29) g/L, and the difference was statistically significant (P<0.05).After observation group patients serum TNF-αand IL-8 were (26.9 ±6.9,12.5 ±3.8) ng/L, than control group (37.5 ±7.4,27.3 ±6.8)ng/L, and the difference was significant (P<0.05).Clinical observation group after treatment colonoscopy was 93.30%, better than control group, 68.89%, and the difference was statistically significant (P<0.05).Conclusion The clinical effect of intestinal Granule more precise treatment of ulcerative colitis, compared with conventional Western medicine has no significant side effects, it is worthy of further research and application.

Objective To explore the effects of maternal allergy symptoms on the cytokine level of umbilical cord blood of neonates.Methods A cross sectional study was conducted among 136 pregnant women in Yiwu city from 1st July to 30th December,in 2012.A questionnaire on social economic charactertistics was used and blood sam-ple of pregnant women and umbilical cord blood sample were collected to detect the level of IgE,eosinophile granulo-cyte,Eotaxin,IL-9,IL-6,IL-4,IL-5,IFN-γ,IL-10.The differences between pregnant women with and without allergy symptoms were carried out.Results There were no significant differences in social and demographic characteristics between the two groups(all P>0.05).Pregnant women with allergy symptoms had higher IgE level(0.13 IU/mL vs 0.10 IU/Ml,Z=-2.063,P=0.039),eosinophile granulocyte(0.39 ×109/mL vs 0.29 ×109/mL,Z=-2.548, P=0.011),Eotaxin(66.18ng/L vs 48.35ng/L,Z=-2.144,P=0.032),IFN-γ(927.81ng/L vs 338.65ng/L,Z=-2.051,P=0.040),IL-10(15.59ng/L vs 11.55ng/L,Z=-2.022,P=0.043) than pregnant women without allergy symptoms in neonates′cord blood.Conclusion Maternal allergy symptoms may increase the level of IgE, eosinophile granulocyte,Eotaxin,IFN-γand IL-10 of neonates′cord blood.

Objective To observe the clinical characteristic and prognosis of primary lung cancer patients with venous thromboembolism (VTE).Methods 589 primary lung cancer patients were selected and divided into VTE group(n =49) and non VTE group(n =540).49 cases with VTE were divided into pulmonary thromboembolism (PTE) group(n =15),including single PTE and PTE combined with deep venous thrombosis(DVT) and DVT group (n =34).Single factor and multiple logistic regression analysis were performed to determine the factors influencing primary lung cancer patients with VTE.Clinical manifestation,time of onset and prognosis of patients with VTE were analyzed.Results 49 patients with VTE included 10 patients(20.4％) with single PTE,34 patients(69.4％) with single DVT and 5 PTE patients combined with DVT(10.2％).D-dimer(OR =1.560,95％ CI =1.018 ～ 2.392,x2 =4.161,P =0.041),interleukin-1 (IL-1,OR =1.846,95％ CI =1.054-3.234,x2 =4.594,P =0.033),tumor necrosis factor (TNF OR =1.486,95％ CI =1.014-2.178,x2 =4.126,P =0.042),adenocarcinoma (OR =2.854,95％CI=1.217-6.695,x2 =5.812,P=0.016) and phase Ⅲ-Ⅳ(OR =2.198,95％CI=1.122-4.305,x2 =5.272,P =0.022) were the factors influencing primary lung cancer patients with VTE.Chest tightness,coughing,accelerated heart rate,swelling and pain in lower limb were common clinical manifestations of primary lung cancer patients with VTE.Most patients with VTE occurred within 3 months after a diagnosis of primary lung cancer.There was no significant difference in the time of onset between PTE group and DVT group(P ＞0.05).As of July 2014,31 cases (63.2％) died,12 cases (24.5 ％) survived,and 6 cases (12.2％) lost in 49 patients with VTE.The median survival time of 49 patients with VTE was 9.5 months.The median survival time of PTE group was 5.8 months,while DVT group was 15.2 months,but no significant difference between them (P ＞ 0.05).Conclusion Increased D-dimer,increased IL-1,increased TNF,adenocarcinoma and phase Ⅲ-Ⅳ could increase the risk of primary lung cancer patients with VTE.There were little typical.clinical symptoms in most patients with VTE,which occurred with in 3 months after a diagnosis of primary lung cancer.They had high mortality and needed to take early diagnosis and treatment through auxiliary examination.

Objective To explorer the clinical effects of Pentoxifylline(PTX group) with mechanical ventilation in patients with acute respiratory distress syndrome(ARDS). Methods 64 patients with ARDS were randomized into a control group(n = 32 ,ARDS conventional treatment) and a PTX group(n = 32 ,PTX 50ml iv bid in addition to the conventional treatment). The patients were closely monitored with their curative effect, blood gas indexes and serum levels of TNF-α and IL-8 after the treatment. Results The PaO_2 and PaO_2/FiO_2 got significantly higher in the PTX group than in the control group on 3d and 7d (P ＜ 0.05). The serum levels of TNF-α and IL-8 and the mortality of ARDS were significantly decreased in the PTX group after the combined with Pentoxifylline (P ＜ 0.05 or P ＜0.01). Conclusion The treatment combined with the Pentoxifylline can effectively improve the Oxygenation and prognosis of ARDS patients by inhibiting the inflammatory responses.

Objective:To prepare and test tetrameric sH-2K~d-HBc complex for the further measurement of the specific CTL response.Methods:PE labled streptavidin with 4 biotinylated binding sites can bind to 4 biotinylated monomer to form the corresponding tetramer.Mice were immunized via different methods of genetic immunization by use of the construted pcDNA3-C plasmid to get the specific CTLs.Then our prepared tetramer was applied to stain the specific CTLs by the analysis of flow cytometry.Results:We applied our prepared tetramer to stain the cells from the experimental groups and control group.The results showed the tetramer was able to discriminate the frequencies of specific CTL induced by the three immunol methods(0.24%,0.26%,0.36% vs 0.07%,P≤0.05).This demonstrated that the prepared tetramer could bind its targets specifically and efficiently.The three immunol methods induced different levels of immune responses.Compared with the traditional muscle injection,gene gun induced weaker humoral immune response and stronger cellular immune response,and hydrodynamic injection induced the strongest humoral and cellular immune responses.Conclusion:Have successfully constructed the sH-2K~d-HBc tetramer.The techniques and methods can be used for preparation of tetramers of other types of MHCⅠ molecules.

This study investigated the expression profiles of IL-10 gene in three human hepatoma cell lines including Huh7, HepG2, and HepG2 transfected with a plasmid containing hepatitis B virus (HBV) named HepG2.2.15. RT-PCR analysis demonstrated that IL-10 message RNA was absent in HepG2 and Huh7 cells, whereas it was present in HepG2.2.15 cells, which was consistent with ELISA result. Furthermore, except for lamivudine other antiviral treatments did not significantly decrease the HBV DNA level in HepG2.2.15 cells, while they had different effects on the expression of IL-10 protein, although stimulation by LPS had no significant effect. In addition, except for poly(I:C), the other treatments decreased the expression of IL-10 protein to different degrees, but had no significant effects on the expression of NF-κB and MyD88. Meanwhile, all treatments we used had effect on the expression of STAT1. In conclusion, IL-10 was expressed in HepG2.2.15 cells and STAT1 pathway might be involved in the regulation of IL-10 expression in HepG2.2.15 cells, but it was not the sole pathway, the exact mechanism warrants further study.

Neural stem/progenitor cells (NSCs) can spontaneously differentiate into neurons and glial cells in the absence of mitogen fibroblast growth factor-2 (FGF-2) or epidermal growth factor (EGF) in medium and the spontaneous differentiation of NSCs is mediated partially by endogenous ciliary neurotrophic factor (CNTF). This study examined the relationship of FGF-2 and CNTF in the spontaneous differentiation of adult hippocampal progenitor cells (AHPs). AHPs were cultured in the medium containing different concentration of FGF-2 (1-100 ng/mL). Western blotting and immunofluorescence staining were applied to detect the expression of the astrocytic marker GFAP, the neuronal marker Tuj1, the oligodendrocytic marker CNPase and, Nestin, the marker of AHPs. The expression of endogenous CNTF in AHPs at early (passage 4) and late stage (passage 22) was also measured by Western blotting. The results showed that FGF-2 increased the expression of Nestin, dramatically inhibited the expression of GFAP and Tuj1 and slightly suppressed the expression of CNPase. FGF-2 down-regulated the expression of endogenous CNTF in AHPs at both early (passage 4) and late stage (passage 22). These results suggested that FGF-2 could inhibit the spontaneous differentiation of cultured AHPs by negatively regulating the expression of endogenous CNTF in AHPs.

Objective To investigate the distribution and sequence conservation of genes encoding the outer membrane lipoprotein D(LPD)of nontypeable Haemophilus influenzae(NTHi)isolates and to ana-lyze the immunogenicity and the immunoprotective effects of the expressed recombinant LPD(rLPD). Meth-ods PCR analysis was used to detect the genes encoding LPD of NTHi isolates. The PCR products were se-quenced after T-A cloning. A prokaryotic expression system for genes encoding LPD was established to ex-press the rLPD. Ni-NTA affinity chromatography was used for purification. SDS-PAGE and Bio-Rad Gel Im-age Analyzer were used to detect the expression and the yield of rLPD. The antigenicity and immunoreactivity of rLPD were detected by ELISA and Western blot assay. The immunoprotective effects of rLPD against lethal dose of NTHi were evaluated in a mouse model. Results All of the tested NTHi isolates were positive for the genes encoding LPD. They shared 98. 0% -99. 4% homologies in nucleotide sequences and 98. 5% -100% homologies in amino acid sequences. The established prokaryotic expression system expressed rLPD with a high yield. High levels of antibody in rabbits were induced by the rLPD. The anti-NTHi antiserum samples from rabbits and children could recognize and react with the rLPD. The result of ELISA indicated that 93. 6%(58 / 62)and 53. 2%(32 / 62)of the serum samples from children with NTHi infection were positive for rLPD-IgM and rLPD-IgG,respectively. The rLPD at concentrations of 100 μg and 200 μg could respectively protect 60. 0% and 73. 3% of mice from lethal NTHi infection. Conclusion The genes enco-ding LPD were extensively distributed in NTHi isolates with high sequence conservation. The expressed rLPD could be used as a potential candidate antigen in the development of genetic engineering vaccine against NTHi infection considering its high immunogenicity and immunoprotective effects.

Hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) are believed to play a major role in viral clearance and disease pathogenesis during HBV infection. To clarify the differences in host immune responses between self-limited and chronic HBV infections, we constructed three HLA-A*0201/HBV tetramers with immunodominant epitopes of core18-27, polymerase 575-583 and envelope 335-343, and analyzed the HBV-specific CTLs in peripheral blood mononuclear cells (PBMCs) from patients infected with HBV. The frequencies and expansion ability of HBV-specific CD8(+) T cells in most self-limited HBV infected individuals were higher than those in chronic HBV-infected patients. HBV-specific CD8(+) T cells could be induced by in vitro peptide stimulation from chronic patients with a low level of serum HBV-DNA but not from those with a high level of serum HBV-DNA. In chronic infection, no significant correlation was found either between the frequencies of HBV-specific CD8(+) T cells and the viral load, or between the frequencies and the levels of alanine transaminase. Our results suggested that the frequencies of HBV-specific CTLs are not the main determinant of immune-mediated protection in chronic HBV infection and immunotherapeutic approaches should be aimed at not only boosting a HBV-specific CD8(+) T response but also improving its function.