The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21–CodonPlus (DE3)–RIL using the pGEX-4T-1 expression vecto...The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21–CodonPlus (DE3)–RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM4B by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD tested serum / OD negative serum≥2.1. The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.展开更多
OBJECTIVE： To explore the regulatory effects of es- tablished Chinese herbal formulas on inflammatory mediators released during asthma attacks, and to elucidate the molecular mechanism of Traditional Chinese Medicine...OBJECTIVE： To explore the regulatory effects of es- tablished Chinese herbal formulas on inflammatory mediators released during asthma attacks, and to elucidate the molecular mechanism of Traditional Chinese Medicine in the treatment of asthma. METHODS： Seventy-five asthmatic children were randomly divided into a Chinese medication group （45 cases） and a Western medication control group （30 cases）. Patients in the Chinese medication group were treated with a series of established Chi- nese herbal formulas, whereas the Western medica- tion control group received a leukotriene receptor antagonist and a bronchial relaxant. Real-time PCR was used to determine the mRNA expression levels of interleukin （IL）-4, cysteinyl leukotriene receptor 1 （CysLTR1）, and interferon （IFN）-γ in peripheralblood mononuclear cells before and after treat- ment. Enzyme-linked immunosorbent assay was used to measure the peripheral blood levels of IL-4, leukotriene （LTE）-4, and INF-γ before and after treat- ment. RESULTS： After treatment, the mRNA expression levels of 11-4 and CysLTR1 were down-regulated （P〈 0.01） and the mRNA expression levels of IFN-γ were up-regulated （P〈0.05） in the Chinese medication and Western medication groups; no significant dif- ference was found between the two groups. In the Chinese medication group, IL-4 blood level was de- creased and it was significantly different from that in the Western medication group （P〈0.05）; there was also a significant increase in IFN-γ blood levels after treatment with Chinese medica- tion （P〈0.05）. There were no significant differenc- es in LTE-4 blood levels between the two groups before and after treatment （P〉0.05）. CONCLUSION： Chinese medication has a regulato- ry effect on leukotriene receptor gene expression and the imbalance of Th1/Th2 immune cells dur- ing asthma attacks in pediatric patients.展开更多
基金Society Commonweal Study of China (2001DIA10006)
文摘The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21–CodonPlus (DE3)–RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM4B by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD tested serum / OD negative serum≥2.1. The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.
基金Supported by Major Research Project of Shanghai Traditional Chinese Medicine Three-year Action Plan(ZYSNXD-CC-ZDYJ034)Shanghai Science and Technology Research Grant Program(No.12401905500)Shanghai Health Bureau Medical Research Fund Grant Program(No.2006L032A)
文摘OBJECTIVE： To explore the regulatory effects of es- tablished Chinese herbal formulas on inflammatory mediators released during asthma attacks, and to elucidate the molecular mechanism of Traditional Chinese Medicine in the treatment of asthma. METHODS： Seventy-five asthmatic children were randomly divided into a Chinese medication group （45 cases） and a Western medication control group （30 cases）. Patients in the Chinese medication group were treated with a series of established Chi- nese herbal formulas, whereas the Western medica- tion control group received a leukotriene receptor antagonist and a bronchial relaxant. Real-time PCR was used to determine the mRNA expression levels of interleukin （IL）-4, cysteinyl leukotriene receptor 1 （CysLTR1）, and interferon （IFN）-γ in peripheralblood mononuclear cells before and after treat- ment. Enzyme-linked immunosorbent assay was used to measure the peripheral blood levels of IL-4, leukotriene （LTE）-4, and INF-γ before and after treat- ment. RESULTS： After treatment, the mRNA expression levels of 11-4 and CysLTR1 were down-regulated （P〈 0.01） and the mRNA expression levels of IFN-γ were up-regulated （P〈0.05） in the Chinese medication and Western medication groups; no significant dif- ference was found between the two groups. In the Chinese medication group, IL-4 blood level was de- creased and it was significantly different from that in the Western medication group （P〈0.05）; there was also a significant increase in IFN-γ blood levels after treatment with Chinese medica- tion （P〈0.05）. There were no significant differenc- es in LTE-4 blood levels between the two groups before and after treatment （P〉0.05）. CONCLUSION： Chinese medication has a regulato- ry effect on leukotriene receptor gene expression and the imbalance of Th1/Th2 immune cells dur- ing asthma attacks in pediatric patients.
