1.Prokaryotic expression of glutathione transferase gene from Taenia solium and its immunological investigations
Daijia LIN ; Huang JIANG ; Niaoxin JIANG ; Jiang NAN ; Wang YUN ; Liuyu JIANG
Chinese Journal of Zoonoses 2010;(2):107-110
In the present study, the prokaryotic expression of glutathione transferase (GST) gene from Taenia solium and its immunological properties were investigated by means of biological informatics methods. The GST gene from T.solium was screened from the cDNA plasmid library of the adult worms. This gene was cloned into prokaryotic expression plasmid pET28a(+) and then expressed in E.coli BL21(DE3) after double enzyme digestion, PCR identification and IPTG induction. The expressed product was identified by SDS-PAGE and the recombinant protein was purified through purification column of His-Ni~(2+) protein. Meanwhile, the immunoreactivity of the purified protein was analyzed by Western blot assay. In these ways, the recombinants were successfully constructed and the highly purified proteins were obtained. It was demonstrated that these proteins could be recognized by sera of patients infected with T.asitica and T.rhynchus saginatus. From these observations, it is evident that highly efficient expression of GST of Taenia solium with definite immunoreactivity can be demonstrated in the prokaryotic expression system.
2.The effect of butylphthalide on expression of VEGF and P38MAPK in hippocampus of Alzheimer's disease rats
Huikai ZHANG ; Xiaowei ZHANG ; Xiaona SHENG ; Liuyu ZHAI ; Guohua ZHANG ; Guoqing JIANG
Journal of Chinese Physician 2013;15(9):1158-1161
Objective To explore the roles of vascular endothelial growth factor (VEGF) and P38MAPK in the pathogenesis of Alzheimer's disease(AD) rats,and the effects of butyIphthalide on the influence of VEGF and P38MAPK in hippocampus of AD rats.Methods SD rats were randomly divided into blank group,AD model group,butylphthalide low-dose group and high dose group (n =8 rats per group).Aggregated Aβ1-42 was injected into the bilateral hippocampus of rats by stereotaxic coordinates method to induce AD.Morris water maze test was used to determine the abilities of learning and memory.Western blotting combined with Gel Doc imagine systems were used to investigate the expression of VEGF and P-P38MAPK in hippocampus of AD rats.Results The result of Morris water maze experiment showed that one week after modeling,escape latency was different(F =66.658,P < 0.05),and either dose the frequency of crossing platform (F =6.884,P <0.05).Compared with the blank group,the other three groups'latent period of escape was extended significantly(P < 0.05),and frequency of crossing platform was significantly less (P < 0.05).After drug intervention for 4 weeks,the expression of VEGF was difference(F =171.064,P <0.05),it was decreased obviously in the model group than blank group(P <=0.05),but it was increased in the drug intervention groups than model group (P < 0.05),and increased more significantly in the high dose group than low dose group (P < 0.05).The expression of P38MAPK had no obvious change among four groups (P > 0.05),however,the expression of P-P38MAPK showed difference(F =104.395,P < 0.05),it was increased in drug intervention and model group than blank group (P < 0.05),it was increased in drug intervention and model group than blank group (P < 0.05),reduced significantly in drug intervention groups than model group (P < 0.05),and decreased more significantly in high dose groups than low dose group (P < 0.05).Conclusions Butylphthalide could obviously enhance the expression of VEGF,reduce the expression of P-P38MAPK in hippocampus of AD rats.