胰腺炎相关蛋白基因（pap）对大鼠肝再生的作用研究
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摘要
胰腺炎相关蛋白(PAP)，是从胰腺炎患者的胰液中发现的一种炎症蛋白，有促进有丝分裂、抗细胞凋亡、抗炎症和促进细胞与胞外基质粘附等作用。为揭示其在肝再生中的作用，本文根据大鼠pap基因的mRNA序列，采用分子克隆的方法克隆出pap基因。在此基础上设计干涉片段并构建出pap基因的两个干涉载体pGenesil-1.0-pap(289)和pGenesil-1.0-pap(485)，三个检验载体pGenesil-1.0-pap(289)-pap、pGenesil-1.0-pap(485)-pap和pGenesil-1.0-HK-pap，并通过尾静脉液压转基因方法检测各检验载体的作用，筛选出最佳干涉载体pGenesil-1.0-pap(485)。采用脂质体转染法将pEGFP-N1-pap及对照pEGFP-N1，pGenesil-1.0-pap(485)及对照pGenesil-1.0-HK等4种质粒转入肝癌细胞BEL-7402内，再用G418筛选出稳定转染细胞株，并进行以下检测：采用活体观察和HE染色观察细胞形态结构；细胞计数和MTT法测定细胞的生长曲线；PCNA细胞免疫化学检测细胞增殖；Hoechst33258染色检测细胞凋亡；甲基纤维素半固体培养基检测集落形成率；药物MTT法检测细胞的耐药性。为进一步研究pap对肝再生的作用奠定基础，也为揭示肝再生的分子机制提供信息。体内实验采用液压转基因技术将pEGFP-N1-pap及对照pEGFP-N1﹑pGenesil-1.0-pap(289)和pGenesil-1.0-pap(485)及对照pGenesil-1.0-HK等5种质粒分别注入2/3肝切除后的大鼠肝脏内，采用常规组织学方法、荧光显微镜技术以及统计学方法等检测转染这些质粒后大鼠的死亡率﹑组织显微结构、肝系数以及肝再生率，以判断pap对肝再生的影响。
体外实验结果表明，转染pEGFP-N1-pap的细胞明显成梭形，细胞核仁增多，与pap融合的绿色荧光蛋白(EGFP)主要分布在细胞核核膜周围，其它转染细胞株形态无明显变化，EGFP主要分布均在细胞核中；生长曲线检测显示转染pEGFP-N1-pap质粒的细胞增殖最快；PCNA细胞免疫化学和hoechst33258染色检测表明稳定转染pEGFP-N1-pap质粒的细胞可能促进细胞增殖、抑制凋亡；集落形成率结果显示稳定转染pEGFP-N1-pap的细胞恶性程度升高；药物MTT检测发现pap基因能促进BEL-7402细胞生长，但对其耐药性无明显影响。体内实验结果表明，转入pEGFP-N1-pap质粒后，有炎症反应发生；在120小时，pEGFP-N1-pap组比对照组的肝系数和肝再生率高，且与对照组相比pEGFP-N1-pap(289)组要比pEGFP-N1-pap(485)组的肝系数和肝再生率高。综上所述，pap基因可能有促进细胞增殖，增加肝癌细胞恶性程度的作用，而对肝癌细胞抗药性没有明显耐药性；pap可能参与促进肝再生进程。
Pancreatitis-associated protein(PAP) is an inflammatory protein what was found in pancreatic juicefrom pancreatitis, to promote mitosis, the anti-apoptotic, anti-inflammatory and promote cell adhesion toextracellular matrix and so on. In order to study its role in liver regeneration, According to the mRNAsequence of rat pap gene, pap gene was cloned and constructed into the vector. On this basis, interferefragments were designed and two interference vectors pGenesil-1.0-pap(289) and pGenesil-1.0-pap(485),three test vectors pGenesil-1.0-pap(289)-pap，pGenesil-1.0-pap(485)-pap and pGenesil-1.0-HK-pap wereestablished, and pap(485) was slected by methods of the tail vein hydraulic testing the role of each vector.The expression vector pEGFP-N1-pap and its control pEGFP-N1, interference vector pGenesil-1.0-pap-(485) and its control pGenesil-1.0-HK transfected into hepatoma cell BEL-7402with lipofectamine, andselection of stably transfected cell lines to the use of G418. Then, all kinds of cell lines above were detectedusing the following methods: cell morphology observed in vivo and by HE staining; cell growth curve bycell counting and MTT assay; cell proliferation detected by PCNA immunocytochemistry; cell apoptosisdetected by Hoechst33258staining; cell colony formation rate detected by Methyl cellulose semi-solidmedium; cell resistance to drugs detected by MTT assay. These laid the foundation for further study of papin liver regeneration and provided information to reveal the mechanism of liver regeneration molecules. Invivo experiments, the pEGFP-N1-pap, pGenesil-1.0-pap(289) and pGenesil-1.0-pap(485) employed thetechnique of hydrodynamics-based transgening were injected into the remnant livers of rat after2/3partialhepatectomy(PH), and the mortality of rats, the morphological structure of regenerating livers, livercoefficient and liver regeneration rate were analyzed using statistical methods.
The results of vitro experiments showed the nucleus of BEL-7402cell transfected with expressionvectors pEGFP-N1-pap significantly increased and looked like spindle; EGFP fusion proteins with papwere mainly distributed around the nucleus nuclear membrane, while no significant changes were observedin other transfected cell lines with EGFP mainly distributed in the nucleus; Growth curve and MTT assayshowed that the proliferation of cells transfected with pEGFP-N1-pap was obvious; PCNAimmunocytochemistry and Hoechst33258staining showed that plasmid pEGFP-N1-pap could promote cell proliferation and inhibit apoptosis; The degree of malignancy of cell lines transfected with pEGFP-N1-papwas increased; MTT assay showed that pap gene can promote BEL-7402cell growth, but no significanteffect on its resistance to drugs. The results of vivo experiments showed that liver coefficient and liverregeneration rate of rats transfected with pEGFP-N1-pap were higher than PH control at120h, but that ofrats transfected with siRNA vectors were lower than the control. In short, pap gene may promote cellproliferation, increase the role of malignancy of hepatoma cell, but has no apparent resistance on hepatomacell as anticancer agents; pap may be involved in promoting the process of liver regeneration.
体外实验结果表明，转染pEGFP-N1-pap的细胞明显成梭形，细胞核仁增多，与pap融合的绿色荧光蛋白(EGFP)主要分布在细胞核核膜周围，其它转染细胞株形态无明显变化，EGFP主要分布均在细胞核中；生长曲线检测显示转染pEGFP-N1-pap质粒的细胞增殖最快；PCNA细胞免疫化学和hoechst33258染色检测表明稳定转染pEGFP-N1-pap质粒的细胞可能促进细胞增殖、抑制凋亡；集落形成率结果显示稳定转染pEGFP-N1-pap的细胞恶性程度升高；药物MTT检测发现pap基因能促进BEL-7402细胞生长，但对其耐药性无明显影响。体内实验结果表明，转入pEGFP-N1-pap质粒后，有炎症反应发生；在120小时，pEGFP-N1-pap组比对照组的肝系数和肝再生率高，且与对照组相比pEGFP-N1-pap(289)组要比pEGFP-N1-pap(485)组的肝系数和肝再生率高。综上所述，pap基因可能有促进细胞增殖，增加肝癌细胞恶性程度的作用，而对肝癌细胞抗药性没有明显耐药性；pap可能参与促进肝再生进程。
Pancreatitis-associated protein(PAP) is an inflammatory protein what was found in pancreatic juicefrom pancreatitis, to promote mitosis, the anti-apoptotic, anti-inflammatory and promote cell adhesion toextracellular matrix and so on. In order to study its role in liver regeneration, According to the mRNAsequence of rat pap gene, pap gene was cloned and constructed into the vector. On this basis, interferefragments were designed and two interference vectors pGenesil-1.0-pap(289) and pGenesil-1.0-pap(485),three test vectors pGenesil-1.0-pap(289)-pap，pGenesil-1.0-pap(485)-pap and pGenesil-1.0-HK-pap wereestablished, and pap(485) was slected by methods of the tail vein hydraulic testing the role of each vector.The expression vector pEGFP-N1-pap and its control pEGFP-N1, interference vector pGenesil-1.0-pap-(485) and its control pGenesil-1.0-HK transfected into hepatoma cell BEL-7402with lipofectamine, andselection of stably transfected cell lines to the use of G418. Then, all kinds of cell lines above were detectedusing the following methods: cell morphology observed in vivo and by HE staining; cell growth curve bycell counting and MTT assay; cell proliferation detected by PCNA immunocytochemistry; cell apoptosisdetected by Hoechst33258staining; cell colony formation rate detected by Methyl cellulose semi-solidmedium; cell resistance to drugs detected by MTT assay. These laid the foundation for further study of papin liver regeneration and provided information to reveal the mechanism of liver regeneration molecules. Invivo experiments, the pEGFP-N1-pap, pGenesil-1.0-pap(289) and pGenesil-1.0-pap(485) employed thetechnique of hydrodynamics-based transgening were injected into the remnant livers of rat after2/3partialhepatectomy(PH), and the mortality of rats, the morphological structure of regenerating livers, livercoefficient and liver regeneration rate were analyzed using statistical methods.
The results of vitro experiments showed the nucleus of BEL-7402cell transfected with expressionvectors pEGFP-N1-pap significantly increased and looked like spindle; EGFP fusion proteins with papwere mainly distributed around the nucleus nuclear membrane, while no significant changes were observedin other transfected cell lines with EGFP mainly distributed in the nucleus; Growth curve and MTT assayshowed that the proliferation of cells transfected with pEGFP-N1-pap was obvious; PCNAimmunocytochemistry and Hoechst33258staining showed that plasmid pEGFP-N1-pap could promote cell proliferation and inhibit apoptosis; The degree of malignancy of cell lines transfected with pEGFP-N1-papwas increased; MTT assay showed that pap gene can promote BEL-7402cell growth, but no significanteffect on its resistance to drugs. The results of vivo experiments showed that liver coefficient and liverregeneration rate of rats transfected with pEGFP-N1-pap were higher than PH control at120h, but that ofrats transfected with siRNA vectors were lower than the control. In short, pap gene may promote cellproliferation, increase the role of malignancy of hepatoma cell, but has no apparent resistance on hepatomacell as anticancer agents; pap may be involved in promoting the process of liver regeneration.
引文
[1] Kirimlioglu V, Kirimlioglu H, Yilmaz S, et al. Effect of fish oil, olive oil, and vitamin E on liverpathology, cell proliferation, and antioxidant defense system in rats subjected to partial hepatectomy[J]. Transplant Proc.2006,38(2):564-567.
[2] Higgins GM, Anderson RM. Experimental pathology of the liver: restoration of the liver of the whiterat following partial surgical removal[J]. Arch Pathol Lab Med.1931,12:186-202.
[3] Taub R. Liver regeneration: from myth to mechanism[J]. Nat Rev Mol Cell Biol.2004,5(10):836-847.
[4] Chan SC, Lo CM, Fan ST. Splanchnic hemodynamics in liver regeneration after right liver livingdonor liver transplantation[J]. Liver Transpl.2010,16(3):412.
[5] Qin SW, Zhao LF, Chen XG, et al. Expression pattern and action analysis of genes associated with theresponses to chemical stimuli during rat liver regeneration[J]. World J Gastroenterol.2006,12(45):7285-7291.
[6] Zhao Q,Ren H,Zhu D,et a1.Stem/progenitor cells in liver injury repair and regeneration[J].B iol Cell.2009,101(10):557-571
[7] Fausto N, Campbell JS, Riehle KJ. Liver regeneration[J]. Hepatology.2006,43(2Suppl1): S45-53.
[8] Reynolds ES. Liver parenchymal cell injury. i. initial alterations of the cell following poisoning withcarbon tetrachloride[J]. J cell boil.1963,19:139-157.
[9] Shinozuka H, Ohmura T, Katyal SL, et al. Possible roles of nonparenchymal cells in hepatocyteproliferation induced by lead nitrate and by tumor necrosis factor alpha[J]. Hepatology.1996,23(6):1572-1577.
[10] Michalopoulos GK, De Frances MC. Liver regeneration[J]. Science.1997,276(5309):60-66.
[11] Zimmermann A. Liver regeneration: the emergence of new pathways[J]. Med Sci Monit.2002,8(3):RA53-63.
[12] Fausto N.Liver regeneration[J]. Hepatol.2000,32(1Suppl):19-31.
[13] Mohammed FF, Khokha R. Thinking outside the cell: proteases regulate hepatocyte division[J].Trends Cell Biol.2005,15(10):555-563.
[14] Bartek J, Lukas J, Bartkova J.Perspective: defects in cell cycle control and cancer[J].J Pathol.1999,187(1):95-99.
[15] Sherr CJ. Cancer cell cycles[J].Science.1996,274(5293):1672-1677.
[16] Albrecht JH, Rieland BM, Nelsen CJ, et al.Regulation of G (1) cyclin-dependent kinases in the liver:role of nuclear localization and p27sequestration[J]. Am J Physiol.1999,277(6Pt1):G1207-1216.
[17] Jaumot M, Estanyol JM, Serratosa J, et al. Activation of cdk4and cdk2during rat liver regenerationis associated with intranuclear rearrangements of cyclin-cdk complexes[J].Hepatology.1999,29(2):385-395.
[18] Menjo M, Ikeda K, Nakanishi M.Regulation of G1cyclin-dependent kinases in liver regeneration[J].J Gastroenterol Hepatol.1998,13(Suppl):S100-105.
[19] Sakamoto T, Liu Z, Murase N,et al. Mitosis and apoptosis in the liver of interleukin-6-deficient miceafter partial hepatectomy[J].Hepatology.1999,29(2):403-411.
[20] Hanse EA, Nelsen CJ, Goggin MM, et al. Cdk2plays a critical role in hepatocyte cell cycleprogression and survival in the setting of cyclin D1expression in vivo[J].Cell Cycle.2009,8(17):2802-2809.
[21] Fausto N, Riehle KJ.Mechanisms of liver regeneration and their clinical implications[J].J Hepatobili-ary Pancreat Surg.2005,12(3):181-189.
[22] Holger W, Amar DS, Arndt V, et al. Loss of p21permits carcinogenesis from chronically damagedliver and kidney epithelial cells despite unchecked apoptosis[J]. Cancer Cell.2008,14(1):59-67.
[23] Starkel P, Lambotteb L, Sempouxc C, et al. After portal branch ligation in the rat, cellularproliferation is associated with selective induction of c-Ha-rss, p53, cyclin E, and cdk2[J]. Gut.2001,49(1):119-130.
[24] Masson S, Harrison DJ, Plevris JN, et al. Potential of hematopoietic stem cell therapy in hepatology:a critical review[J].Stem Cells.2004,22(6):897-907.
[25] Hanashiro K, Kanai M, Geng Y, et al. Roles of cyclins A and E in induction of centrosomeamplification in p53-compromised cells[J].Oncogene.2008,27(40):5288-5302.
[26] Hunter T, Pines J. Cyclins and cancer. II: Cyclin D and CDK inhibitors come of age[J].Cell.1994,79(4):573-582.
[27] Kato A, Ota S, Bamba H, et al. Regulation of cyclin D-dependent kinase activity in rat liverregeneration[J].Biochem Biophys Res Commun.1998,245(1):70-74.
[28] Albrecht JH, Poon RY, Ahonen CL, et al. Involvement of p21and p27in the regulation of CDKactivity and cell cycle progression in the regenerating liver[J]. Oncogene.1998,16(16):2141–2150.
[29] Hirai H, Roussel MF, Kato JY, et al.Novel INK4proteins, p19and p18, are specific inhibitors of thecyclin D-dependent kinases CDK4and CDK6[J].Mol Cell Biol.1995,15(5):2672-2681.
[30] Harper JW, Elledge SJ, Keyomarsi K, et al. Inhibition of cyclin-dependent kinases by p21[J]. MolBiol Cell.1995,6(4):387-400.
[31] Lu Z, Hunter T. Ubiquitylation and proteasomal degradation of the p21(Cip1), p27(Kip1) andp57(Kip2) CDK inhibitors[J].Cell Cycle.2010,9(12):2342-2352.
[32] Cheng M, Olivier P, Diehl JA, et al.The p21(Cip1) and p27(Kip1) CDK 'inhibitors' are essentialactivators of cyclin D-dependent kinases in murine fibroblasts[J].EMBO J.1999,18(6):1571-1583.
[33] Karavias DD, Tsamandas AC, Tepetes K, et al. BCL-2and BAX expression and cell proliferationafter partial hepatectomy with and without ischemia on cholestatic liver in rats:an experimentalstudy[J]. J Surg Res.2003,110(2):399-408.
[34] Noji S, Tashiro K, Koyama E et al. Expression of hepatocyte growth factor gene in endothelial andKupffer cells of damaged rat livers, as revealed by in situ hybridization[J]. Biochem. Biophys. Res.Commun.1990,173(1):42-47.
[35] Depoortere F, Pirson I, Bartek J, et al.Transforming growth factor beta(1) selectively inhibits thecyclic AMP-dependent proliferation of primary thyroid epithelial cells by preventing the associationof cyclin D3-cdk4with nuclear p27(kip1)[J].Mol Biol Cell.2000,11(3):1061-1076.
[36] Dixon M, Agius L, Yeaman SJ, Inhibition of rat hepatocyte proliferation by transforming growthfactor beta and glucagon is associated with inhibition of ERK2and p70S6kinase[J].Hepatology.1999,29(5):1418-1424.
[37] Yamada Y, Kirillova I, Peschon JJ, et al. Initiation of liver growth by tumor necrosis factor: deficientliver regeneration in mice lacking type I tumor necrosis factor receptor[J].Proc Natl Acad Sci USA.1997,94(4):1441-1446.
[38] Iavarone A, Massagué J.Repression of the CDK activator Cdc25A and cell-cycle arrest by cytokineTGF-beta in cells lacking the CDK inhibitor p15[J].Nature.1997,387(6631):417-422.
[39] Poon RY, Toyoshima H, Hunter T.Redistribution of the CDK inhibitor p27between differentcyclin.CDK complexes in the mouse fibroblast cell cycle and in cells arrested with lovastatin orultraviolet irradiation[J].Mol Biol Cell.1995,6(9):1197-1213.
[40] Keim V, Rohr G, Stocker HG, et al. An additional secretory protein in the rat pancreas[J].Digestion.1984,29(4):242-249.
[41] Katsumata N, Chakraborty C, Myal Y, et al. Molecular cloning and expression of peptide23,a growthhormonereaeasing hormone-inducible pituitary[J]. Endocrinology.1995,136(4):1332-1339.
[42] Okamoto H.The Reg gene family and Reg proteins: with special attention to the regeneration ofpancreatic beta-cells[J].J Hepatobiliary Pancreat Surg.1999,6(3):254-262.
[43] Abe M, Nata K, Akiyama T, et al.Identification of a novel Reg family gene, Reg IIIdelta, andmapping of all three types of Reg family gene in a75kilobase mouse genomic region[J]. Gene.2000,246(1-2):111-122.
[44] Keim V, Iovanna JL, Rohr G, et al. Characterization of a rat pancreatic secretory protein associatedwith pancreatitis[J]. Gastroenterology.1991,100(3):775-782.
[45] Dusetti NJ, Ortiz EM, Mallo GV,et al. Pancreatitis-associated protein I (PAP I), an acute phaseprotein induced by cytokines. Identification of two functional interleukin-6response elements in therat PAP I promoter region[J].J Biol Chem.1995,270(38):22417-22421.
[46] Iovanna JL, Keim V, Bosshard A,et al.PAP, a pancreatic secretory protein induced during acutepancreatitis, is expressed in rat intestine[J].Am J Physiol.1993,265(4Pt1):G611-618.
[47] Yoshiharu M, Yoshitake S, Ikurou M, et al.Serum levels of pancreatitis-assoceated protein indigestive diseases with special reference to gast rointestinal ancers[J]. Dig Dis Sci.1999,44(6):1142-1147.
[48] Chakraborty C, Vrontakis M, Molnar P, et al. Expression of pituitary peptide23in the rat uterus:regulation by estradiol[J]. Mol Cell Endocrinol.1995,108(1-2):149-154.
[49] Chakraborty C, Sharma S, Katsumata N, et al. Plasma clearance, tissue uptake and expression ofpituitary peptide23/pancreatitis-associated protein in the rat[J].J Endocrinol.1995,145(3):461-469.
[50] Chakraborty C, Katsumata N, Myal Y, et al.Age-related changes in peptide-23/pancreatitis-associat-ed protein and pancreatic stone protein/reg gene expression in the rat and regulation by growthhormone-releasing hormone[J].Endocrinology.1995,136(5):1843-1849.
[51] B deker H, Keim V, Fiedler F,et al. PAP I interacts with itself, PAP II, PAP III, andlithostathine/regIalpha[J].Mol Cell Biol Res Commun.1999,2(3):150-154.
[52] Zhang YW, Ding LS, Lai MD.Reg gene family and human diseases[J].World J Gastroenterol.2003,9(12):2635-2641.
[53] Iovanna J, Orelle B, Keim V,et al. Messenger RNA sequence and expression of ratpancreatitis-associated protein,a lectin-related protein overexpressed during acute experimentalpancreatitis[J].J Biol Chem.1991,266(36):24664-24669.
[54] Lasserre C, Simon MT, Ishikawa H, et al.Structural organization and chromosomal localization of ahuman gene (HIP/PAP) encoding a C-type lectin overexpressed in primary liver cancer[J].Eur JBiochem.1994,224(1):29-38.
[55] Dusetti NJ, Frigerio JM, Fox MF,et al.Molecular cloning, genomic organization, and chromosomallocalization of the human pancreatitis-associated protein (PAP) gene[J].Genomics.1994,19(1):108-114.
[56] Simon MT, Pauloin A, Normand G,et al. HIP/PAP stimulates liver regeneration after partialhepatectomy and combines mitogenic and anti-apoptotic functions through the PKA signalingpathway[J].FASEB J.2003,17(11):1441-1450.
[57] Terazono K, Yamamoto H, Takasawa S,et al.A novel gene activated in regenerating islets[J].J BiolChem.1988,263(5):2111-2114.
[58] Harada K, Zen Y, Kanemori Y,et al. Human REG I gene is up-regulated in intrahepatic cholangiocar-cinoma and its precursor lesions[J].Hepatology.2001,33(5):1036-1042.
[59] Yamaoka T, Yoshino K, Yamada T,et al.Diabetes and tumor formation in transgenic mice expressingReg I[J].Biochem Biophys Res Commun.2000,278(2):368-376.
[60] White P, Brestelli JE, Kaestner KH, et al.Identification of transcriptional networks during liverregeneration[J].J Biol Chem.2005,280(5):3715-3722.
[61] Yuan RH, Jeng YM, Chen HL,et al. Opposite roles of human pancreatitis-associated protein andREG1A expression in hepatocellular carcinoma: association of pancreatitis-associated proteinexpression with low-stage hepatocellular carcinoma, beta-catenin mutation, and favorable prognosis[J].Clin Cancer Res.2005,11(7):2568-2575.
[62] Macadam RC, Sarela AI, Farmery SM,et al.Death from early colorectal cancer is predicted by thepresence of transcripts of the REG gene family[J].Br J Cancer.2000,83(2):188-195.
[63] Lasserre C, Christa L, Simon MT, et al.A novel gene (HIP) activated in human primary liver cancer[J].Cancer Res.1992,52(18):5089-5095.
[64] Dusetti NJ, Montalto G, Ortiz EM, et al. Mechanism of PAP I gene induction duringhepatocarcinogenesis: clinical implications[J].Br J Cancer.1996,74(11):1767-1775.
[65] Yokoyama Y, Nagino M, Nimura Y. Mechanisms of hepatic regeneration following portalveinembolization and partial hepatectomy: a review[J]. World J Surg.2007,31(2):367-374.
[66] Ma XM, Dong XS, Zhao HL. Research progress of liver regeneration-related molecules[J]. Interna-tional J Surg,2009,36(11):760-763.
[67] Kountouras J, Boura P, Lygidakis NJ. Liver regeneration after hepatectomy[J]. Hepatogastroenterol-ogy.2001,48(38):556-562.
[68]张春艳．L-4-氟苯丙氨酸等6种氨基酸衍生物的生物学作用研究[D]．河南：河南师范大学，2011．
[69]杨莹．L-硝基-4-苯丙氨酸等四种氨基酸衍生物的生物活性检测[D]．河南：河南师范大学，2011．
[70] Moniaux N, Song H, Darnaud M,et al.Human hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein cures fas-induced acute liver failure in mice by attenuating free-radical damage ininjured livers[J].Hepatology.2011,53(2):618-627.
[71]梁博，程世翔，涂悦等．SNCG真核表达载体构建及其在胶质瘤细胞中的稳定表达和影响[J]．细胞与分子免疫学杂志．2011，27(4)：412-414．
[72]孔祥平，郑国池，张宜俊，等．肝细胞刺激因子活性的实验研究[J]．天津医药．1989，17(7)：401-404．
[73]邱财荣，李新田，龙超良，等．四甲基偶氮唑蓝比色法测定促肝细胞生长素对SMMC-7721细胞的作用[J]．中国医院药学杂志．2006，26(1)：42-44．
[74]王林，郑书深，董福生，等．PCNA在成釉细胞瘤中表达的免疫组化研究[J]．现代口腔医学杂志．2004，18(2)：188-189．
[75] Pack RA, Hardy K, Madigan MC, et al.Differential effects of the antioxidant alpha-lipoic acid on theproliferation of mitogen-stimulated peripheral blood lymphocytes and leukaemic T cells[J].MolImmunol.2002,38(10):733-745.
[76] Takasawa S,lkeda T,Akiyama T,et a1．Cyclin D1activation through ATF-2in Reg-induced pancreaticbeta-cell regeneration[J]. FEBS Lett.2006,580(2):585-591.
[77]郭昱，郭霞，姚金峰，等．黄芩甙对肝癌细胞BEL-7402形态学的影响[J]．中国组织化学与细胞化学杂志．2008，17(2)：174-179．
[78]朱秀敏．细胞凋亡的形态特征与分子机制概述[J]．生物学教学．2011，36(11)：6-8．
[79] Lu Y，Ponton A，Okamoto H，et a1．Activation of the Reg family genes by pancreatic-specific IGF-Igene deficiency and after streptozotocin-induced diabetes in mouse pancreas[J]．Am J PhysiolEndocrinol Metab.2006,291(1):E50-58.
[80] Zenilman ME, Tuchman D, Zheng Q, et al. Comparison of reg I and reg III levels during acutepancreatitis in the rat[J]. Ann Surg.2000,232(5):646-652.
[81] Xu C, Chen X, Chang C, et al. Transcriptome analysis of hepatocytes after partial hepatectomy inrats[J]. Dev Genes Evol,2010,220(9-10):263-274
[82] Michalopoulos GK. Liver regeneration after partial hepatectomy: critical analysis of mechanisticdilemmas[J]. Am J Pathol,2010,176(1):2-13.
[83] Chijiiwa K, Nakano K, Kameoka N, et al. Proliferating cell nuclear antigen, plasma fibronectin, andliver regeneration rate after seventy percent hepatectomy in normal and cirrhotic rats. Surgery[J].1994,116(3):544-549.
[84] Yang X, Jin H, Liu K, et al. A Novel Peptide Derived from Human Pancreatitis-Associated ProteinInhibits Inflammation In Vivo and In Vitro and Blocks NF-Kappa B Signaling Pathway[J]. PLoS One.2011,6(12):e29155.
[85] Yang J, Jiang H, Chen SS, et al. Lentivirus-mediated RNAi targeting CREB binding proteinattenuates neointimal formation and promotes re-endothelialization in balloon injured rat carotidartery[J]. Cell Physiol Biochem,2010,26(3):441-448.
[86]刘晓艳，方红，陈鸿超，等．CMV启动子调控的VEGF-shRNA表达载体的构建及有效干扰序列的筛选[J]．浙江大学学报(医学版)．2010，39(2)：181-186．
[87]孙静，魏从文，管楷，等．人TBK1小干扰RNA质粒的构建及稳定干扰TBK1细胞株的筛选[J]．生物技术通讯．2010，21(2)：163-167．
[88]邢雪琨，徐存拴，张富春，等．一种快速筛选有效RNAi片段的方法[J]．生物技术通讯．2010,20(6)：46-49．
[89]徐存拴，邢雪琨，谢来峰，等．液压转基因技术应用于大鼠肝脏转基因研究[J]．解剖学报．2009，40(1)：103-107．
[90] Wolff JA，Malone RW, Williams P, et al. Direct gene transfer into mouse muscle in vivo[J]. Science.1990,247(4949pt1):1465-1468.
[91] Liu F, Song Y, Liu D. Hydrodynamics-based transfection in animals by systemic administration ofplasmid DNA[J]. Gene Ther.1999,6(7):1258-1266.
[92] Zhang G, Budker V, Wolff JA. High levels of foreign gene expression in hepatocytes after tail veininjections of naked plasmid DNA[J]. Hum Gene Ther.1999,10(10):1735-1737.
[93]徐存拴，邢雪琨，杨献光，等．液压转基因技术应用于大鼠再生肝转基因实验[J]．解剖学报．2009，40(4)；599-603．
[94] Chen ZY, Yant SR, Lie CY, et al. Linear DNAs concatemerize in vivo and result in sustainedtransgene expression in mouse liver[J]. Mol Ther.2001,3(3):403-410.
[95]陈栋，吴力群，曹景玉，等．大鼠肝切除术后肝损伤程度与肝再生状态的动态对比研究[J]．中华肝胆外科杂志．2002，8(6)；354-357．
[96]贺晨霞，吴文君，丁友法，等．尾静脉大容量快速注射法介导的人凝血因子Ⅸ基因在小鼠肝内的高效表达[J]．科学通报．2003，48(5)：447-451．
[97] Sakamoto T, Liu Z, Murase N,et al. Mitosis and apoptosis in the liver of interleukin-6-deficient miceafter partial hepatectomy[J].Hepatology.1999,29(2):403-411.
[98] Uzan B，Figeac F，Portha B，et a1．Mechanisms of KGF mediatedsignaling in pancreatic duct cellproliferation and differentiation[J]．PLoS One.2009,4(3):e4734.
[99] Motoo Y，Satomura Y，Moufi I，et a1．Serum levels of pancreatitis-associated protein in digestivediseases with special reference to gastrointestinal cancers[J]．Dig Dis Sci.1999,44(6):1142-1147.
[2] Higgins GM, Anderson RM. Experimental pathology of the liver: restoration of the liver of the whiterat following partial surgical removal[J]. Arch Pathol Lab Med.1931,12:186-202.
[3] Taub R. Liver regeneration: from myth to mechanism[J]. Nat Rev Mol Cell Biol.2004,5(10):836-847.
[4] Chan SC, Lo CM, Fan ST. Splanchnic hemodynamics in liver regeneration after right liver livingdonor liver transplantation[J]. Liver Transpl.2010,16(3):412.
[5] Qin SW, Zhao LF, Chen XG, et al. Expression pattern and action analysis of genes associated with theresponses to chemical stimuli during rat liver regeneration[J]. World J Gastroenterol.2006,12(45):7285-7291.
[6] Zhao Q,Ren H,Zhu D,et a1.Stem/progenitor cells in liver injury repair and regeneration[J].B iol Cell.2009,101(10):557-571
[7] Fausto N, Campbell JS, Riehle KJ. Liver regeneration[J]. Hepatology.2006,43(2Suppl1): S45-53.
[8] Reynolds ES. Liver parenchymal cell injury. i. initial alterations of the cell following poisoning withcarbon tetrachloride[J]. J cell boil.1963,19:139-157.
[9] Shinozuka H, Ohmura T, Katyal SL, et al. Possible roles of nonparenchymal cells in hepatocyteproliferation induced by lead nitrate and by tumor necrosis factor alpha[J]. Hepatology.1996,23(6):1572-1577.
[10] Michalopoulos GK, De Frances MC. Liver regeneration[J]. Science.1997,276(5309):60-66.
[11] Zimmermann A. Liver regeneration: the emergence of new pathways[J]. Med Sci Monit.2002,8(3):RA53-63.
[12] Fausto N.Liver regeneration[J]. Hepatol.2000,32(1Suppl):19-31.
[13] Mohammed FF, Khokha R. Thinking outside the cell: proteases regulate hepatocyte division[J].Trends Cell Biol.2005,15(10):555-563.
[14] Bartek J, Lukas J, Bartkova J.Perspective: defects in cell cycle control and cancer[J].J Pathol.1999,187(1):95-99.
[15] Sherr CJ. Cancer cell cycles[J].Science.1996,274(5293):1672-1677.
[16] Albrecht JH, Rieland BM, Nelsen CJ, et al.Regulation of G (1) cyclin-dependent kinases in the liver:role of nuclear localization and p27sequestration[J]. Am J Physiol.1999,277(6Pt1):G1207-1216.
[17] Jaumot M, Estanyol JM, Serratosa J, et al. Activation of cdk4and cdk2during rat liver regenerationis associated with intranuclear rearrangements of cyclin-cdk complexes[J].Hepatology.1999,29(2):385-395.
[18] Menjo M, Ikeda K, Nakanishi M.Regulation of G1cyclin-dependent kinases in liver regeneration[J].J Gastroenterol Hepatol.1998,13(Suppl):S100-105.
[19] Sakamoto T, Liu Z, Murase N,et al. Mitosis and apoptosis in the liver of interleukin-6-deficient miceafter partial hepatectomy[J].Hepatology.1999,29(2):403-411.
[20] Hanse EA, Nelsen CJ, Goggin MM, et al. Cdk2plays a critical role in hepatocyte cell cycleprogression and survival in the setting of cyclin D1expression in vivo[J].Cell Cycle.2009,8(17):2802-2809.
[21] Fausto N, Riehle KJ.Mechanisms of liver regeneration and their clinical implications[J].J Hepatobili-ary Pancreat Surg.2005,12(3):181-189.
[22] Holger W, Amar DS, Arndt V, et al. Loss of p21permits carcinogenesis from chronically damagedliver and kidney epithelial cells despite unchecked apoptosis[J]. Cancer Cell.2008,14(1):59-67.
[23] Starkel P, Lambotteb L, Sempouxc C, et al. After portal branch ligation in the rat, cellularproliferation is associated with selective induction of c-Ha-rss, p53, cyclin E, and cdk2[J]. Gut.2001,49(1):119-130.
[24] Masson S, Harrison DJ, Plevris JN, et al. Potential of hematopoietic stem cell therapy in hepatology:a critical review[J].Stem Cells.2004,22(6):897-907.
[25] Hanashiro K, Kanai M, Geng Y, et al. Roles of cyclins A and E in induction of centrosomeamplification in p53-compromised cells[J].Oncogene.2008,27(40):5288-5302.
[26] Hunter T, Pines J. Cyclins and cancer. II: Cyclin D and CDK inhibitors come of age[J].Cell.1994,79(4):573-582.
[27] Kato A, Ota S, Bamba H, et al. Regulation of cyclin D-dependent kinase activity in rat liverregeneration[J].Biochem Biophys Res Commun.1998,245(1):70-74.
[28] Albrecht JH, Poon RY, Ahonen CL, et al. Involvement of p21and p27in the regulation of CDKactivity and cell cycle progression in the regenerating liver[J]. Oncogene.1998,16(16):2141–2150.
[29] Hirai H, Roussel MF, Kato JY, et al.Novel INK4proteins, p19and p18, are specific inhibitors of thecyclin D-dependent kinases CDK4and CDK6[J].Mol Cell Biol.1995,15(5):2672-2681.
[30] Harper JW, Elledge SJ, Keyomarsi K, et al. Inhibition of cyclin-dependent kinases by p21[J]. MolBiol Cell.1995,6(4):387-400.
[31] Lu Z, Hunter T. Ubiquitylation and proteasomal degradation of the p21(Cip1), p27(Kip1) andp57(Kip2) CDK inhibitors[J].Cell Cycle.2010,9(12):2342-2352.
[32] Cheng M, Olivier P, Diehl JA, et al.The p21(Cip1) and p27(Kip1) CDK 'inhibitors' are essentialactivators of cyclin D-dependent kinases in murine fibroblasts[J].EMBO J.1999,18(6):1571-1583.
[33] Karavias DD, Tsamandas AC, Tepetes K, et al. BCL-2and BAX expression and cell proliferationafter partial hepatectomy with and without ischemia on cholestatic liver in rats:an experimentalstudy[J]. J Surg Res.2003,110(2):399-408.
[34] Noji S, Tashiro K, Koyama E et al. Expression of hepatocyte growth factor gene in endothelial andKupffer cells of damaged rat livers, as revealed by in situ hybridization[J]. Biochem. Biophys. Res.Commun.1990,173(1):42-47.
[35] Depoortere F, Pirson I, Bartek J, et al.Transforming growth factor beta(1) selectively inhibits thecyclic AMP-dependent proliferation of primary thyroid epithelial cells by preventing the associationof cyclin D3-cdk4with nuclear p27(kip1)[J].Mol Biol Cell.2000,11(3):1061-1076.
[36] Dixon M, Agius L, Yeaman SJ, Inhibition of rat hepatocyte proliferation by transforming growthfactor beta and glucagon is associated with inhibition of ERK2and p70S6kinase[J].Hepatology.1999,29(5):1418-1424.
[37] Yamada Y, Kirillova I, Peschon JJ, et al. Initiation of liver growth by tumor necrosis factor: deficientliver regeneration in mice lacking type I tumor necrosis factor receptor[J].Proc Natl Acad Sci USA.1997,94(4):1441-1446.
[38] Iavarone A, Massagué J.Repression of the CDK activator Cdc25A and cell-cycle arrest by cytokineTGF-beta in cells lacking the CDK inhibitor p15[J].Nature.1997,387(6631):417-422.
[39] Poon RY, Toyoshima H, Hunter T.Redistribution of the CDK inhibitor p27between differentcyclin.CDK complexes in the mouse fibroblast cell cycle and in cells arrested with lovastatin orultraviolet irradiation[J].Mol Biol Cell.1995,6(9):1197-1213.
[40] Keim V, Rohr G, Stocker HG, et al. An additional secretory protein in the rat pancreas[J].Digestion.1984,29(4):242-249.
[41] Katsumata N, Chakraborty C, Myal Y, et al. Molecular cloning and expression of peptide23,a growthhormonereaeasing hormone-inducible pituitary[J]. Endocrinology.1995,136(4):1332-1339.
[42] Okamoto H.The Reg gene family and Reg proteins: with special attention to the regeneration ofpancreatic beta-cells[J].J Hepatobiliary Pancreat Surg.1999,6(3):254-262.
[43] Abe M, Nata K, Akiyama T, et al.Identification of a novel Reg family gene, Reg IIIdelta, andmapping of all three types of Reg family gene in a75kilobase mouse genomic region[J]. Gene.2000,246(1-2):111-122.
[44] Keim V, Iovanna JL, Rohr G, et al. Characterization of a rat pancreatic secretory protein associatedwith pancreatitis[J]. Gastroenterology.1991,100(3):775-782.
[45] Dusetti NJ, Ortiz EM, Mallo GV,et al. Pancreatitis-associated protein I (PAP I), an acute phaseprotein induced by cytokines. Identification of two functional interleukin-6response elements in therat PAP I promoter region[J].J Biol Chem.1995,270(38):22417-22421.
[46] Iovanna JL, Keim V, Bosshard A,et al.PAP, a pancreatic secretory protein induced during acutepancreatitis, is expressed in rat intestine[J].Am J Physiol.1993,265(4Pt1):G611-618.
[47] Yoshiharu M, Yoshitake S, Ikurou M, et al.Serum levels of pancreatitis-assoceated protein indigestive diseases with special reference to gast rointestinal ancers[J]. Dig Dis Sci.1999,44(6):1142-1147.
[48] Chakraborty C, Vrontakis M, Molnar P, et al. Expression of pituitary peptide23in the rat uterus:regulation by estradiol[J]. Mol Cell Endocrinol.1995,108(1-2):149-154.
[49] Chakraborty C, Sharma S, Katsumata N, et al. Plasma clearance, tissue uptake and expression ofpituitary peptide23/pancreatitis-associated protein in the rat[J].J Endocrinol.1995,145(3):461-469.
[50] Chakraborty C, Katsumata N, Myal Y, et al.Age-related changes in peptide-23/pancreatitis-associat-ed protein and pancreatic stone protein/reg gene expression in the rat and regulation by growthhormone-releasing hormone[J].Endocrinology.1995,136(5):1843-1849.
[51] B deker H, Keim V, Fiedler F,et al. PAP I interacts with itself, PAP II, PAP III, andlithostathine/regIalpha[J].Mol Cell Biol Res Commun.1999,2(3):150-154.
[52] Zhang YW, Ding LS, Lai MD.Reg gene family and human diseases[J].World J Gastroenterol.2003,9(12):2635-2641.
[53] Iovanna J, Orelle B, Keim V,et al. Messenger RNA sequence and expression of ratpancreatitis-associated protein,a lectin-related protein overexpressed during acute experimentalpancreatitis[J].J Biol Chem.1991,266(36):24664-24669.
[54] Lasserre C, Simon MT, Ishikawa H, et al.Structural organization and chromosomal localization of ahuman gene (HIP/PAP) encoding a C-type lectin overexpressed in primary liver cancer[J].Eur JBiochem.1994,224(1):29-38.
[55] Dusetti NJ, Frigerio JM, Fox MF,et al.Molecular cloning, genomic organization, and chromosomallocalization of the human pancreatitis-associated protein (PAP) gene[J].Genomics.1994,19(1):108-114.
[56] Simon MT, Pauloin A, Normand G,et al. HIP/PAP stimulates liver regeneration after partialhepatectomy and combines mitogenic and anti-apoptotic functions through the PKA signalingpathway[J].FASEB J.2003,17(11):1441-1450.
[57] Terazono K, Yamamoto H, Takasawa S,et al.A novel gene activated in regenerating islets[J].J BiolChem.1988,263(5):2111-2114.
[58] Harada K, Zen Y, Kanemori Y,et al. Human REG I gene is up-regulated in intrahepatic cholangiocar-cinoma and its precursor lesions[J].Hepatology.2001,33(5):1036-1042.
[59] Yamaoka T, Yoshino K, Yamada T,et al.Diabetes and tumor formation in transgenic mice expressingReg I[J].Biochem Biophys Res Commun.2000,278(2):368-376.
[60] White P, Brestelli JE, Kaestner KH, et al.Identification of transcriptional networks during liverregeneration[J].J Biol Chem.2005,280(5):3715-3722.
[61] Yuan RH, Jeng YM, Chen HL,et al. Opposite roles of human pancreatitis-associated protein andREG1A expression in hepatocellular carcinoma: association of pancreatitis-associated proteinexpression with low-stage hepatocellular carcinoma, beta-catenin mutation, and favorable prognosis[J].Clin Cancer Res.2005,11(7):2568-2575.
[62] Macadam RC, Sarela AI, Farmery SM,et al.Death from early colorectal cancer is predicted by thepresence of transcripts of the REG gene family[J].Br J Cancer.2000,83(2):188-195.
[63] Lasserre C, Christa L, Simon MT, et al.A novel gene (HIP) activated in human primary liver cancer[J].Cancer Res.1992,52(18):5089-5095.
[64] Dusetti NJ, Montalto G, Ortiz EM, et al. Mechanism of PAP I gene induction duringhepatocarcinogenesis: clinical implications[J].Br J Cancer.1996,74(11):1767-1775.
[65] Yokoyama Y, Nagino M, Nimura Y. Mechanisms of hepatic regeneration following portalveinembolization and partial hepatectomy: a review[J]. World J Surg.2007,31(2):367-374.
[66] Ma XM, Dong XS, Zhao HL. Research progress of liver regeneration-related molecules[J]. Interna-tional J Surg,2009,36(11):760-763.
[67] Kountouras J, Boura P, Lygidakis NJ. Liver regeneration after hepatectomy[J]. Hepatogastroenterol-ogy.2001,48(38):556-562.
[68]张春艳．L-4-氟苯丙氨酸等6种氨基酸衍生物的生物学作用研究[D]．河南：河南师范大学，2011．
[69]杨莹．L-硝基-4-苯丙氨酸等四种氨基酸衍生物的生物活性检测[D]．河南：河南师范大学，2011．
[70] Moniaux N, Song H, Darnaud M,et al.Human hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein cures fas-induced acute liver failure in mice by attenuating free-radical damage ininjured livers[J].Hepatology.2011,53(2):618-627.
[71]梁博，程世翔，涂悦等．SNCG真核表达载体构建及其在胶质瘤细胞中的稳定表达和影响[J]．细胞与分子免疫学杂志．2011，27(4)：412-414．
[72]孔祥平，郑国池，张宜俊，等．肝细胞刺激因子活性的实验研究[J]．天津医药．1989，17(7)：401-404．
[73]邱财荣，李新田，龙超良，等．四甲基偶氮唑蓝比色法测定促肝细胞生长素对SMMC-7721细胞的作用[J]．中国医院药学杂志．2006，26(1)：42-44．
[74]王林，郑书深，董福生，等．PCNA在成釉细胞瘤中表达的免疫组化研究[J]．现代口腔医学杂志．2004，18(2)：188-189．
[75] Pack RA, Hardy K, Madigan MC, et al.Differential effects of the antioxidant alpha-lipoic acid on theproliferation of mitogen-stimulated peripheral blood lymphocytes and leukaemic T cells[J].MolImmunol.2002,38(10):733-745.
[76] Takasawa S,lkeda T,Akiyama T,et a1．Cyclin D1activation through ATF-2in Reg-induced pancreaticbeta-cell regeneration[J]. FEBS Lett.2006,580(2):585-591.
[77]郭昱，郭霞，姚金峰，等．黄芩甙对肝癌细胞BEL-7402形态学的影响[J]．中国组织化学与细胞化学杂志．2008，17(2)：174-179．
[78]朱秀敏．细胞凋亡的形态特征与分子机制概述[J]．生物学教学．2011，36(11)：6-8．
[79] Lu Y，Ponton A，Okamoto H，et a1．Activation of the Reg family genes by pancreatic-specific IGF-Igene deficiency and after streptozotocin-induced diabetes in mouse pancreas[J]．Am J PhysiolEndocrinol Metab.2006,291(1):E50-58.
[80] Zenilman ME, Tuchman D, Zheng Q, et al. Comparison of reg I and reg III levels during acutepancreatitis in the rat[J]. Ann Surg.2000,232(5):646-652.
[81] Xu C, Chen X, Chang C, et al. Transcriptome analysis of hepatocytes after partial hepatectomy inrats[J]. Dev Genes Evol,2010,220(9-10):263-274
[82] Michalopoulos GK. Liver regeneration after partial hepatectomy: critical analysis of mechanisticdilemmas[J]. Am J Pathol,2010,176(1):2-13.
[83] Chijiiwa K, Nakano K, Kameoka N, et al. Proliferating cell nuclear antigen, plasma fibronectin, andliver regeneration rate after seventy percent hepatectomy in normal and cirrhotic rats. Surgery[J].1994,116(3):544-549.
[84] Yang X, Jin H, Liu K, et al. A Novel Peptide Derived from Human Pancreatitis-Associated ProteinInhibits Inflammation In Vivo and In Vitro and Blocks NF-Kappa B Signaling Pathway[J]. PLoS One.2011,6(12):e29155.
[85] Yang J, Jiang H, Chen SS, et al. Lentivirus-mediated RNAi targeting CREB binding proteinattenuates neointimal formation and promotes re-endothelialization in balloon injured rat carotidartery[J]. Cell Physiol Biochem,2010,26(3):441-448.
[86]刘晓艳，方红，陈鸿超，等．CMV启动子调控的VEGF-shRNA表达载体的构建及有效干扰序列的筛选[J]．浙江大学学报(医学版)．2010，39(2)：181-186．
[87]孙静，魏从文，管楷，等．人TBK1小干扰RNA质粒的构建及稳定干扰TBK1细胞株的筛选[J]．生物技术通讯．2010，21(2)：163-167．
[88]邢雪琨，徐存拴，张富春，等．一种快速筛选有效RNAi片段的方法[J]．生物技术通讯．2010,20(6)：46-49．
[89]徐存拴，邢雪琨，谢来峰，等．液压转基因技术应用于大鼠肝脏转基因研究[J]．解剖学报．2009，40(1)：103-107．
[90] Wolff JA，Malone RW, Williams P, et al. Direct gene transfer into mouse muscle in vivo[J]. Science.1990,247(4949pt1):1465-1468.
[91] Liu F, Song Y, Liu D. Hydrodynamics-based transfection in animals by systemic administration ofplasmid DNA[J]. Gene Ther.1999,6(7):1258-1266.
[92] Zhang G, Budker V, Wolff JA. High levels of foreign gene expression in hepatocytes after tail veininjections of naked plasmid DNA[J]. Hum Gene Ther.1999,10(10):1735-1737.
[93]徐存拴，邢雪琨，杨献光，等．液压转基因技术应用于大鼠再生肝转基因实验[J]．解剖学报．2009，40(4)；599-603．
[94] Chen ZY, Yant SR, Lie CY, et al. Linear DNAs concatemerize in vivo and result in sustainedtransgene expression in mouse liver[J]. Mol Ther.2001,3(3):403-410.
[95]陈栋，吴力群，曹景玉，等．大鼠肝切除术后肝损伤程度与肝再生状态的动态对比研究[J]．中华肝胆外科杂志．2002，8(6)；354-357．
[96]贺晨霞，吴文君，丁友法，等．尾静脉大容量快速注射法介导的人凝血因子Ⅸ基因在小鼠肝内的高效表达[J]．科学通报．2003，48(5)：447-451．
[97] Sakamoto T, Liu Z, Murase N,et al. Mitosis and apoptosis in the liver of interleukin-6-deficient miceafter partial hepatectomy[J].Hepatology.1999,29(2):403-411.
[98] Uzan B，Figeac F，Portha B，et a1．Mechanisms of KGF mediatedsignaling in pancreatic duct cellproliferation and differentiation[J]．PLoS One.2009,4(3):e4734.
[99] Motoo Y，Satomura Y，Moufi I，et a1．Serum levels of pancreatitis-associated protein in digestivediseases with special reference to gastrointestinal cancers[J]．Dig Dis Sci.1999,44(6):1142-1147.