1.Construction of a DNA vaccine encoding herpes simplex virus type 1 glycoprotein D
Jianguo LI ; Yukang YUAN ; Jie FENG ; Hanqing XU ; Lin SHI
Journal of Xinxiang Medical College 2002;19(3):153-157
Objective To construct a recombinant plasmid DNA containing herpes simplex virus type 1(HSV-1) glycoprotein D (gD) gene.Methods The HSV-1 gD gene was obtained by polymerase chain reaction (PCR) and inserted into TA cloning vector pGEM-T, then cloned into the eukaryotic expression vector pcDNA3.1 to generate pLy-D. The recombinant plasmid pLy-D, which was confirmed by partial sequencing and restriction endonuclease analysis, was transfected into Cos-7 cells and used to inoculate ICR mice via muscular injection. Immunohistochemistry and enzyme-linked immunoabsorbent assay (ELISA) were employed to test the gD expression in transfected cells and the specific anti-HSV-1 antibody in the serum of immunized mice, respectively.Results The gD eukaryotic expression plasmid pLy-D was constructed. Using the immunohistochemistry technique, the gD expression in pLy-D-transfected cells was detected. The ELISA demonstrated that specific anti-HSV-1 antibody could be induced in immunized mice after three times injection.Conclusions We constructed HSV-1 gD eukaryotic expression plasmid pLy-D which could express gD protein in transfected cells and could induce humoral immune response in mice. This observation will be helpful in designing HSV prophylactic vaccine.
2.Primary study on immunomodulatory effect of ursolic acid in mice
Yingjie FENG ; Huixun REN ; Yukang YUAN ; Fei SHEN ; Guixiang FAN
Journal of Xi'an Jiaotong University(Medical Sciences) 2003;0(06):-
Objective To confirm the immunomodulatory effect of ursolic acid on normal and immunosuppressive mice caused by cyclophosphamide.Methods We assayed the function of T lymphocyte transformation with MTT test,examined the function of B lymphocyte with hemolysis spectrophotography,determined the function of non-specific immunity with phagocytosis of macrophages,and measured the content changes of Th_(1) cytokine IL-2 and Th_(2) cytokine IL-6 in serum with ELISA.Results Ursolic acid had a suppressant effect on the function of humoral immunity and enhanced cell-mediated immune response at the low dosage but presented opposite effect at the high dosage in normal mice.Ursolic acid significantly improved immune function on immunosuppressive mice caused by cyclophosphamide(P
3.Application of Fuzzy Comprehensive Evaluation in Formulation Screening
Longlong LI ; Pengbin HAO ; Wenpeng WANG ; Yukang FENG ; Yunping QU ; Guozhi JIANG ; Zhenjiang LI
China Pharmacy 2015;(28):3957-3959,3960
OBJECTIVE:To investigate the application of fuzzy comprehensive evaluation in formulation screening. METH-ODS:Taking Compound paracetamol and chlorphenamine maleate granules as model drug,based on single factor experiment,L9(34) orthogonal test was used to screen the formulation with the amount of aspartame,hawthorn powder essence,milk powder essence and saccharose as factors,using dissolution rate of caffeine as index. The granules prepared by 9 kinds of formulation in orthogonal test were evaluated with fuzzy comprehensive evaluation in terms of taste,aroma,color and dissolubility. The optimized formula-tion was validated and compared with original formulation. RESULTS:The formulation 5 was the optimal choice in both orthogo-nal test and fuzzy comprehensive evaluation,and obtained same results. Compared with sensory comprehensive score (63.12) and dissolution rate of caffeine (91.3%) in original formulation,two indicators of optimized formulation in 3 tests were 84.00 and 99.07% in average (RSD<2.0%,n=3). CONCLUSIONS:Fuzzy comprehensive evaluation can be used for the formulation screening of sensory evaluation index. It is scientific and accurate.