项目已按期完成了预定实验。构建了pEGFP-N2/Pim-3质粒及其激酶失活突变体，证实了通过流体力学方法可实现目的基因的动物活体肝靶向性转移；建立了暴发性肝衰竭（FHF）动物模型，证明细胞凋亡是急性肝衰竭的主要病理形态学特征，而凋亡的发生在早期与iNOS、中晚期与p53基因表达有关，且至少三条凋亡通路参与了该病理过程。在此基础上，研究了Pim-3基因的肝保护效应。研究证实，Pim-3基因活体肝转移明显提高了FHF动物的生存率，减轻了肝组织的炎性损伤，并抑制了肝凋亡和caspase-3活性。其机制为Pim-3基因抑制了炎性细胞因子的表达和分泌、抑制了p53和iNOS基因表达，而促进了抗凋亡蛋白Bcl-2表达。项目的成功实施也促进了Pim-3基因在其它组织损伤修复中的研究，已发现外源Pim-3基因的应用可有效保护肠粘膜上皮细胞内毒素性损伤和心肌细胞缺血/再灌注损伤等。

In this experimental, pEGFP-N2/Pim-3 plasmid and its mutant were successfully constructed, and hepatic target transfer of intrest gene was finished by hydrodynamics-based procedure in vivo. Fulminant hepatic failur was induced by challenge with LPS and D-galactosamine in rats. It is suggested that, during liver failure, the main morphological feature is hepatic apoptosis, which may be associated with a high expression of iNOS (early period) and p53 genes (in the mid and late stages) and at least three apoptosis pathways participate in the pathogenesis. In addition, exogenous Pim-3 was found to protect rats against LPS/D-GalN-induced lethality by improving inflammatory response of liver tissues and inhibiting liver apoptosis and caspase-3 activity, which could be associated with inhibiting secret of inflammatory mediators and promoting expression of anti-apoptosis protein Bcl-2.

项目已按期完成了预定实验。构建了pEGFP-N2/Pim-3质粒及其激酶失活突变体，证实了通过流体力学方法可实现目的基因的动物活体肝靶向性转移；建立了暴发性肝衰竭（FHF）动物模型，证明细胞凋亡是急性肝衰竭的主要病理形态学特征，而凋亡的发生在早期与iNOS、中晚期与p53基因表达有关，且至少三条凋亡通路参与了该病理过程。在此基础上，研究了Pim-3基因的肝保护效应。研究证实，Pim-3基因活体肝转移明显提高了FHF动物的生存率，减轻了肝组织的炎性损伤，并抑制了肝凋亡和caspase-3活性。其机制为Pim-3基因抑制了炎性细胞因子的表达和分泌、抑制了p53和iNOS基因表达，而促进了抗凋亡蛋白Bcl-2表达。项目的成功实施也促进了Pim-3基因在其它组织损伤修复中的研究，已发现外源Pim-3基因的应用可有效保护肠粘膜上皮细胞内毒素性损伤和心肌细胞缺血/再灌注损伤等。