| This Ph.D dissertation is composed of four parts. (1) cloning and analysis of up-regulated expressed genes in rat hepatic reneneration following short interval successive partial hepatectomy; (2) construction of cDNA library following short interval successive partial hepatectomy; (3) clone full-lengh cDNA of differentially expressed genes following short interval successive partial hepatectomy. (4) detection of different phase and apoptosis related proteins in rat hepatic regeneration following short interval successive partial hepatectomy.Liver is the only organ that possesses the great capability of regeneration in mammals. Since Higgins and Anderson established the model of partial hepatectomy (PH) in 1931. it has been widely used to study the reconstructing and regeneration of tissues and organs, cellular multiplication, dedifferentiation, stress response and the regulation of physiology and biochemistry. But due to the complexity of hepatic regeneration, the organ-specific molecular mechanism in hepatic regeneration remains to be elucidated. In order to shed light on the mechanism of hepatic regeneration, Xu Cunshuan et al established the model of short interval successive partial hepatectomy (SISPH) model in the world which had an interval of 4h (cell enter into dedifferentiation stage) and/or 36h (the peak of cell division). With the two crucial point being control, The model had widely been focused and approbated by domestic and outland professinals.In the study, we used SISPH as model, resorting to SSH supression subtractive Hybridization) technique, constrction of cDNA library, Southern blot and Western blot et al. we studied the differentially expressed ESTs (expressed sequence tags ) between control and the materials following short interval successive partial hepatectomy and up-regulated expressed full-length genes and expressed proteins in cell cycle and apoptosis in different phases. Materials and methods:We adopted SISPH as model, sham as control and 112h hepatic tissue as treatment, to study the up-regulated expressed genes following SISPH; I12h hepatic tissue as experimental material, recurring to RT-PCR, restriction endonucleases digestion, proteases digestion, protein package to construct cDNA library; cloning full-length genes by Southern blot from cDNA library; detection proteins in cell cycle and apoptosis proteins in different phases by Western blot. Results:1 x We have cloned 9 ESTs which was 100% homologous to GenBank and 44 ESTs which were homologous to GenBank, one of these ESTs was found novel gene in GenBank. The 53 up-regulated expressed ESTs mainly contained the following categories:(1) Related to positive/negative major acute phase protein (MAP) mRNA genes. Such as serum amyloid A 2, transferrin, haptoglobin, alpha acid glyprotein, flbrinogen like factor which may have closely relation to hepatic regeneration;(2) Relatd to mitochondrial oxidative phosphorylation genes. Rattus norvegicus ankyrin protein,Rattus norvegicus mitochondrial cytochrome oxidase subunits I, II, III genes, ATPase subunit 6 gene, Trp-,Ala-,Asn-,Cys-. Tyr-, Ser (ucn)-, Asp-, Lys-, ly-, Arg-, His-, Ser (agy)-, Leu (cun)-tRNAs which connected with mitochondrial functional compensory prolification due to hypoxia in partial hepatic tissue, they also suggested liver regeneration demand more energy; The EST homologous to Bos taurus mitochondrial NADH: ubiquinone oxidoreductase (complex I) 23-kD subunit (CI-23kD) showed the pathway depended on NADH (malate and glutamic acid) may play important role in hepatic regeneration;(3) Related to protein synthesis genes. The ESTs 95% homologous to Mus musculus mitochondrial ribosomal protein 63 (Mrp63) showed large quantities of rRNA synthesis may meet hepatic regeneration;(4) Related to cell division genes. Microtubulin associated protein (MAP) is not only the target of signal transduction but also the target of an important engine molecular cdc-2 kinase which could regulate cell cycles; MAP also participate...
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