Aniso-dose glycyrrhiza polysaccharide was injected into abdominal cavity of mice. The effect of glycyrrhiza polysaccharide on growth performance and immune ruction of mice was determined. The results showed that the g...Aniso-dose glycyrrhiza polysaccharide was injected into abdominal cavity of mice. The effect of glycyrrhiza polysaccharide on growth performance and immune ruction of mice was determined. The results showed that the growth performance and immune function of mice were improved and there were significant differences among the treatment groups and control group.展开更多
AIM:Interferon α2b (IFNα2b) and thymosin α1 (Tα1) exhibit synergic effects in the treatment of hepatitis B and hepatitis C when used together. For developing a fusion protein drug, fusion proteins of IFNα2b and T...AIM:Interferon α2b (IFNα2b) and thymosin α1 (Tα1) exhibit synergic effects in the treatment of hepatitis B and hepatitis C when used together. For developing a fusion protein drug, fusion proteins of IFNα2b and Ta1 linked by different lengths of (G4S)n(n = 1-3) were constructed and expressed in Pichia pastoris. METHODS: Using PCR and molecular clone techniques, the fusion genes of IFNα2b-(G4S)n-Tα1 (n = 1-3) were constructed and subcloned into the eukaryotic expression vector pPIC9. After transformation of these plasmids into P. pastoris, the expressed fusion proteins IFNα2b-(G4S) n-Tα1 (n = 1-3) were obtained. These proteins were purified through diethylaminoethyl (DEAE) affinity chromatography and Superdex?75 gel filtration and analyzed by SDS-PAGE and Western blot. Antiviral and E-rosette assays were used to investigate the bioactivities of these fusion proteins. RESULTS: DNA sequencing confirmed that the fusion genes of IFNa2b-(G4S)n-Tα1 (n= 1-3) were correctly cloned to the pPIC9 vector. The recombinant IFNα2b-(G4S)n-Tα1 (n = 1-3) fusion proteins expressed in P. pastoris were purified with DEAE and Superdex?75 gel filtration chromatography. The fusion proteins could be observed on sodium dodecylsulfate-polyacrylamide gel electrophoresis with molecular weight (MW) of 23.2, 22.9, and 22.6 ku, respectively, and reacted to the IFNa2b monoclonal antibody and Tal polyclonal antibody. The purified fusion proteins exhibit antiviral activity and can enhance the percentage of E-rosette-forming-cell in E-rosette assay. CONCLUSION: The recombinant IFNa2b-(G4S)n-Tα1 (n = 1-3) fusion proteins were successfully expressed in P. pastoris. Purified fusion proteins exhibit both antiviral activity of IFNa2b and immunomodulatory activity of Tal in vitro. These results will be the basis for further evaluation of the fusion proteins' function in vivo.展开更多
In order to study the mechanism of immunopotentiator, the quantity of T lymphocyte was observed. Total 240 1-day chikens were divided into 3 groups randomly： one contrast group and two groups being drunk immunopotent...In order to study the mechanism of immunopotentiator, the quantity of T lymphocyte was observed. Total 240 1-day chikens were divided into 3 groups randomly： one contrast group and two groups being drunk immunopotentiator according to the concentration 10 mL·L^-1 and 5 mL·L^-1 lasting for 48 d. The number of T lymphocyte in blood was measured by E-rosette when the chikens were at 12-, 24-, 36-, and 48-day. The results showed that the percent ofT lymphocyte in the trial group was obviously higher than that of the contrast, and the 10 mL·L^-1 group was higher than that of the 5 mL·L^-1 group; the quantity distribution of T lymphocyte in intestinal mucosa lymphoid tissue of 14-day, 21-day chiken was surveyed in the contrast group and the10 mL·L^-1 group with the method of routine histology-slices and ANAE, the results showed that the quantity of T lymphocyte in the 10 mL·L^-1 group was significantly higher than that of the contrast, which indicated that immunopotentiator increased markedly the quantity of T lymphocyte, and the 10 mL·L^-1 group was higher than that of the 5 mL·L^-1 group.展开更多
基金Natural Science Foundation of Henan Educational Committee(2003230110)Key Research Foundation of Henan Institute of Science and Techonology~~
文摘Aniso-dose glycyrrhiza polysaccharide was injected into abdominal cavity of mice. The effect of glycyrrhiza polysaccharide on growth performance and immune ruction of mice was determined. The results showed that the growth performance and immune function of mice were improved and there were significant differences among the treatment groups and control group.
文摘AIM:Interferon α2b (IFNα2b) and thymosin α1 (Tα1) exhibit synergic effects in the treatment of hepatitis B and hepatitis C when used together. For developing a fusion protein drug, fusion proteins of IFNα2b and Ta1 linked by different lengths of (G4S)n(n = 1-3) were constructed and expressed in Pichia pastoris. METHODS: Using PCR and molecular clone techniques, the fusion genes of IFNα2b-(G4S)n-Tα1 (n = 1-3) were constructed and subcloned into the eukaryotic expression vector pPIC9. After transformation of these plasmids into P. pastoris, the expressed fusion proteins IFNα2b-(G4S) n-Tα1 (n = 1-3) were obtained. These proteins were purified through diethylaminoethyl (DEAE) affinity chromatography and Superdex?75 gel filtration and analyzed by SDS-PAGE and Western blot. Antiviral and E-rosette assays were used to investigate the bioactivities of these fusion proteins. RESULTS: DNA sequencing confirmed that the fusion genes of IFNa2b-(G4S)n-Tα1 (n= 1-3) were correctly cloned to the pPIC9 vector. The recombinant IFNα2b-(G4S)n-Tα1 (n = 1-3) fusion proteins expressed in P. pastoris were purified with DEAE and Superdex?75 gel filtration chromatography. The fusion proteins could be observed on sodium dodecylsulfate-polyacrylamide gel electrophoresis with molecular weight (MW) of 23.2, 22.9, and 22.6 ku, respectively, and reacted to the IFNa2b monoclonal antibody and Tal polyclonal antibody. The purified fusion proteins exhibit antiviral activity and can enhance the percentage of E-rosette-forming-cell in E-rosette assay. CONCLUSION: The recombinant IFNa2b-(G4S)n-Tα1 (n = 1-3) fusion proteins were successfully expressed in P. pastoris. Purified fusion proteins exhibit both antiviral activity of IFNa2b and immunomodulatory activity of Tal in vitro. These results will be the basis for further evaluation of the fusion proteins' function in vivo.
文摘In order to study the mechanism of immunopotentiator, the quantity of T lymphocyte was observed. Total 240 1-day chikens were divided into 3 groups randomly： one contrast group and two groups being drunk immunopotentiator according to the concentration 10 mL·L^-1 and 5 mL·L^-1 lasting for 48 d. The number of T lymphocyte in blood was measured by E-rosette when the chikens were at 12-, 24-, 36-, and 48-day. The results showed that the percent ofT lymphocyte in the trial group was obviously higher than that of the contrast, and the 10 mL·L^-1 group was higher than that of the 5 mL·L^-1 group; the quantity distribution of T lymphocyte in intestinal mucosa lymphoid tissue of 14-day, 21-day chiken was surveyed in the contrast group and the10 mL·L^-1 group with the method of routine histology-slices and ANAE, the results showed that the quantity of T lymphocyte in the 10 mL·L^-1 group was significantly higher than that of the contrast, which indicated that immunopotentiator increased markedly the quantity of T lymphocyte, and the 10 mL·L^-1 group was higher than that of the 5 mL·L^-1 group.
