AIM:To look for a suitable signal peptide which may effectively conduct hepatopoietin(HPO)secretion,various recombinant eukaryotic expression vectors were constructed.METHODS:Different exogenous signal sequences were spliced with HPO cDNA by PCR,and the spliced genes were cloned into eukaryotic expression plasmids.The different recombinants were respectively tansfected into COS-7 cells by Lipofectamine 2000 method and the secretion of HPO was analyzed by Western blotting.RESULTS:Western blotting analysis indicated that the signal peptides from interleukin-1 receptor antagonist(IL-1ra)and an artifical signal peptide did not secret HPO directly and effectively,but the signal peptide from murine Ig kappa secreted HPO directly with great efficiency.The molecular weight of the secreted HPO was 30 kD,which means that the secreted HPO existed in homodimer.CONCLUSION:Secreted recombinant expression plasmid is successful constructed.The result may pave the way for the gene therapy of hepatic fibrosis.

AIM:To explore the effects of the new drug of sulfonylurea(1-{4-[2-(3-ethyl-4-methyl-2-oxo-3-pyrroline-1-carboxamido)ethyl]-phenylsulfonyl}-3-(1,4-tetramethylene)-urea,BGW) on the glucose uptake and the activation of Akt/PKB in SMMC7721 cells.METHODS:Cultured SMMC7721 cells were divided into control group,glibenclamide group,insulin group,BGW group and BGW+insulin group.Scintillation was used to detect the glucose uptake in SMM7721 cells.The activation of Akt/PKB was tested by Western blotting.RESULTS:Compared to control cells,gibenclamide,insulin,BGW and BGW+insulin significantly increased the glucose uptake(P