Effect of Ershen Zeshu Mixture on the peritoneal lymphatic stomata and the ascites drainage / 中国病理生理杂志

AIM: To further study the effect of Ershen Zeshu Mixture, a Chinese medicine, on the peritoneal lymphatic stomata and the ascites drainage in experimental mice model of CCl 4-induced fibrosis.By the application of NO supplier (sodium nitroprusside,SNP) and NO suppressor (N G-mononethyl-L-arginine,L-NMMA),the effect of NO on the peritoneal lymphatic stomata and the ascites drainage was studied to clarify the relationship between Ershen Zeshu Mixture and NO in the regulation of the peritoneal lymphatic stomata.METHODS: The mouse model of fibrosis was established with the application of intragastric installations of carbon tetrachloride one time every three days;Scanning electron microscope and computer image processing were used to detect the girth,area and distribution density of the peritoneal lymphatic stomata;the concentration of urinary ions was measured in the experiment.RESULTS: ①Compared to NS group and model group,Ershen Zeshu Mixture could significantly increase the girth and the area of the peritoneal lymphatic stomata,promote its distribution density ( P

HIGH-LEVEL EXPRESSION OF RECOMBINANT TRICHOSANTHIN IN ESCHERICHIA COLI AND EVALUATION OF ITS IMMUNOGENICITY / 解剖学报

Objective The TCS gene without N-and C-terminal sequences was cloned into pET-29b in order to get the recombinant trichosanthin(rTCS) at the relatively high level in E.coli,and the rTCS immunogenicity was compared with natural TCS(nTCS). Methods TCS gene without the N-and C-terminal coding sequences was amplified by RT-PCR,and then inserted into pET-29b.The soluble rTCS was induced with IPTG and purified through metal chelate affinity chromatography,the purity and content were showed by the thin-layer scan analysis and bradford assay.After the rabbits were immunized with purified rTCS and nTCS,respectively,the titer and specificity of polyclonal antibody were detected by indirect ELISA assay and Western blotting. Results 1.The vector pET-29b-TCS was constructed successfully after it was confirmed with the same restriction enzymes.2.The rTCS attained the maximum in the presence of 1 mmol/L IPTG for 4?h at 30℃.3.The soluble rTCS was expressed at relatively high level(36% of total protein in E.coli) and the yield of rTCS with a purity of 95% was 1.2?g in 1 L bacterium.4.The polyclonal antisera titer of nTCS was significantly higher than that of rTCS(1∶40?960 and 1∶5?120 respectively).5.The specific reaction between rTCS and anti-nTCS antibody was detected by Western blotting.Conclusion The high level expressing vector pET-29b-TCS,carrying TCS gene withoug the N-and C-terminal coding sequences,was constructed successfully.The immunogenicity of rTCS was remarkably lower than that of nTCS,which indicates that glycosylation of nTCS has a relationship with the antigenicity.