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Objective Hilar cholangiocarcinoma (HC) is invariably fatal without surgical resection. The primary aim of the current study was to determine the safety of variable surgical resections for patient with HC and their survival after surgical resection. In addition, prognostic factor for the overall survival was also evaluated. Methods The study included 59 consecutive patients who were newly diagnosed with HC and underwent surgical resections with curative intend between February 2009 and February 2017. Patients were followed up at 3-6 months intervals after hospital discharge. Postoperative complications and overall survival were determined. Associations of clinicopathologic and surgeon-related factors with overall survival were evaluated through univariate analysis and Cox regression analysis. Results Of patients with Bismuth and Corlette (B & C) type Ⅲ (=19) and Ⅳ (=25) HC lesions, 33 (55.9%) were treated with hilar resection combined with major liver resection (MLR), while the other 11 patients with type Ⅲ and Ⅳ, and those with type Ⅰ (=8) and Ⅱ (=7) HC lesions were treated with hilar resection. The overall surgical mortality was 5.1% and surgical morbidity was 35.6%. There was no statistical difference in the mortality between MLR group and hilar resection group (6.1% 3.8%; =0.703, =0.145). The median follow-up period was 18 months (range, 1-94 months). The 1-, 3-, 5-year survival rate was 59.3%, 36.5%, and 17.7%, respectively. The overall survival after resections was 18 months. In HC patients with B & C type Ⅲ and Ⅳ lesions, the median survival was 23 months for hilar resection with MLR and 8 months for hilar resection alone; the 1-, 3-, 5-year cumulative survival rate was 63.9%, 23.3%, and 15.5%, respectively for hilar resection with MLR, and 11.1%, 0, and 0, respectively for hilar resection alone, with significant differene observed (, 9.902; 95% , 2.636-19.571, =0.001). Four factors were independently associated with overall survival: preoperative serum Ca19-9 (, 7.039; 95% , 2.803-17.678, <0.001), histopathologic grade (, 4.964; 95% , 1.046-23.552, =0.044), surgical margins (=0.031), and AJCC staging (=0.015). Conclusions R0 resection is efficacious in surgical treatment of HC. MLR in combination with caudate lobe resection may increase the chance of R0 resection and improve survival of HC patients with B & C type Ⅲ and Ⅳ lesions. Preoperatively prepared for biliary drainage may ensure the safety of MLR in most HC patients. Novel adjuvant therapies are needed to improve the survival of HC patients with poor prognostic factors.
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<p><b>BACKGROUND</b>Overexpression of G-protein coupled receptor 34 (GPR34) affects the progression and prognosis of human gastric adenocarcinoma, however, the role of GPR34 in gastric cancer development and progression has not been well-determined. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.</p><p><b>METHODS</b>The expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.</p><p><b>RESULTS</b>The ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).</p><p><b>CONCLUSIONS</b>GPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.</p>
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Humanos , Apoptose , Genética , Fisiologia , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células , Genética , Fisiologia , RNA Interferente Pequeno , Genética , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Lisofosfolipídeos , Genética , Metabolismo , Neoplasias Gástricas , Genética , MetabolismoRESUMO
To study the effects of Smad4 on liver fibrosis and hepatocarcinogenesis in mice treated with CCl(4)/ethanol. The wild-type mice (Smad4 +/+) and the Smad4 knockout mice (Smad4 +/-) were injected subcutaneously with carbon tetrachloride(CCl(4))/ethanol twice a week for twenty weeks. The expression of Smad4, TGFbeta1, Smad2, Smad3, Smad6, TIMP1, MMP2 and MMP9 was detected by RT-PCR. In the cirrhotic liver, the expression of Smad4 mRNA was significantly higher than that in the normal liver. Comparing with wild-type mice (Smad4 +/+), the TGFbeta1-Smad4 signaling was markedly attenuated in the Smad4 knockout mice (Smad4 +/-). After induction by CCl(4)/ethanol, the hepatic fibrosis in the Smad4 knockout mice (Smad4 +/-) was obviously alleviated compared with the wild-type mice (Smad4 +/+), and the incidence rate of hepatocarcinogenesis of the former was also lower than that of the latter(32.0% vs 41.9%). These results indicate that knocking out Smad4 can delay the progression of liver fibrosis and liver cancer.
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Animais , Feminino , Masculino , Camundongos , Tetracloreto de Carbono , Modelos Animais de Doenças , Etanol , Cirrose Hepática Experimental , Metabolismo , Patologia , Neoplasias Hepáticas Experimentais , Metabolismo , Patologia , Camundongos Knockout , RNA Mensageiro , Genética , Metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Proteínas Smad , Genética , Metabolismo , Proteína Smad4 , Genética , Metabolismo , Inibidor Tecidual de Metaloproteinase-1 , Genética , Metabolismo , Fator de Crescimento Transformador beta1 , Genética , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To observe the effects of galectin-3 on proliferation and apoptosis of hepatic stellate cells.</p><p><b>METHODS</b>RT-PCR and Western blot were used to detect the expression of galectin-3 in hepatic stellate cells. Short hairpin DNA targeting galectin-3 of rat was was ligated into the recombinant vector pGCsilencer U6/Neo/GFP/shRNA plasmid. Then the plasmid was transfected into rat hepatic stellate cells. RT-PCR and Western blot were used to detect the interfering efficiency. Cell proliferation level was observed by CCK8 method at 24, 48 and 72 hours after transfection. Cell apoptosis was measured by Annexin V/PI-labeled flow cytometric analysis.</p><p><b>RESULTS</b>Expression of galectin-3 in HSC was verified by both RT-PCR and Western blot. The recombinant vector was successfully constructed and verified, and was transfected into rat hepatic stellate cells. Western Blot and RT-PCR results demonstrated that the expression level of Galectin-3 was significantly down-regulated in galectin-3 shRNA transfected cells compared to control vector transferred cells. CCK8 assay indicated that proliferation of Galectin-3 knockdown cells was lower than that of control cells 48 and 72 hours post-transfection. Apoptotic cells in shRNA-interfering group were higher than those in control group both in early stage and advanced stage.</p><p><b>CONCLUSION</b>Hepatic stellate cells can express galectin-3. Inhibition of galectin-3 using RNAi technique can suppress proliferation and induce apoptosis in HSC.</p>
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Animais , Ratos , Apoptose , Linhagem Celular , Proliferação de Células , Regulação para Baixo , Citometria de Fluxo , Galectina 3 , Genética , Metabolismo , Vetores Genéticos , Células Estreladas do Fígado , Biologia Celular , Metabolismo , Cirrose Hepática , Patologia , Plasmídeos , Genética , Interferência de RNA , RNA Mensageiro , Genética , Metabolismo , RNA Interferente Pequeno , Genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , TransfecçãoRESUMO
NY-ESO-1 is an important member of cancer-testis antigen family and is widely distributed among many cancer types. As a tumor-specific antigen with the strongest immunogenicity so far identified, it can induce spontaneous antibody and T-cell responses in patients with NY-ESO-1-positive tumors. Therefore, it has been a good vaccine candidate in the immunotherapy against many malignancies. This article reviews the recent research advances in NY-ESO-1 and its relevant vaccines.
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Humanos , Antígenos de Neoplasias , Genética , Alergia e Imunologia , Usos Terapêuticos , Vacinas Anticâncer , Alergia e Imunologia , Usos Terapêuticos , Ensaios Clínicos como Assunto , Imunoterapia , Proteínas de Membrana , Genética , Alergia e Imunologia , Usos Terapêuticos , Neoplasias , Genética , Alergia e Imunologia , TerapêuticaRESUMO
<p><b>OBJECTIVE</b>To identify a naturally presented HLA-A2-restricted epitope of MAGE-A3 antigen, FLWGPRALV (MAGE-A3(271 - 279)), on the surface of a human hepatocellular carcinoma (HCC) cell line HLE.</p><p><b>METHODS</b>Synthetic peptide FLWGPRALV, served as positive control target, was analyzed by HPLC and HPLC-ESI-TOF-MSMS, in order to determine its HPLC elution time, mass-spectrometric characteristics and the lowest detection limitation by the two approaches. 3 x 10(9) HLE cells were collected, peptides naturally presented by major histocompatibility complex (MHC) molecules on the cell surface were isolated by mild acid elution, and concentrated by lyophilization, then the mixtures of peptides were fractioned by HPLC. The ingredient ranged from 2 min before the elution time determined by the synthetic peptide to 2 min after that was collected, concentrated by lyophilization, and analyzed by HPLC-ESI-TOF-MSMS, to identify the existence of the MAGE-A3(271 - 279) peptide.</p><p><b>RESULTS</b>The HPLC-ESI-TOF-MSMS detection provided an evidence for the existence of a doubly charged ion of (m/z)(2) 529.9, which was further analyzed by collision induced dissociation. The doubly charged ion was ultimately identified as the MAGE-A3(271 - 279) peptide, its amino sequence was FLWGPRALV and its molecular weight was 1058.4 Da.</p><p><b>CONCLUSIONS</b>MAGE-A3(271 - 279) epitope could be naturally presented by HLA-A2 molecules to the surface of HCC cell line and MAGE-A3(271 - 279) peptide may have potential immunotherapeutic value in HCC patients.</p>
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Humanos , Sequência de Aminoácidos , Apresentação de Antígeno , Antígenos de Neoplasias , Carcinoma Hepatocelular , Alergia e Imunologia , Patologia , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Epitopos de Linfócito T , Antígeno HLA-A2 , Alergia e Imunologia , Neoplasias Hepáticas , Alergia e Imunologia , Patologia , Espectrometria de Massas , Proteínas de NeoplasiasRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of protein kinase C (PKC)/transforming growth factor beta 1 (TGF beta1) pathway on activation of hepatic stellate cells (HSC).</p><p><b>METHODS</b>HSC rHSC-99 cell line was used in three groups in this study. Group A served as a control. In group B the HSC were incubated with PKC agonist PMA (0.5 micromol/L), and in group C the cells were incubated with PKC inhibitor calphostin C (100 nmol/L). The PKC activities were detected at different incubation time points (0, 3, 6, 12 and 24 h). Western blot and RT-PCR were used to detect the expression of TGF beta1, Smad 4, collagen type I, III and alpha-smooth muscle actin (alpha-SMA) at the 24 h point. Cell proliferation was assessed by MTT colorimetric assay.</p><p><b>RESULTS</b>PMA increased the activity of PKC significantly, whereas calphostin C inhibited the activity of PKC. The increased activity of PKC promoted the HSC to express TGF beta1, Smad 4, collagen type I, III and alpha-SMA. In comparison with the controls, the expressions of TGF beta1, Smad 4, collagen type I, III and alpha-SMA increased 4.8, 13.1, 2.4, 1.8 and 1.3 fold respectively (P < 0.01). PKC promoted the proliferation of HSC. The above effects were inhibited by the inhibition of PKC activity.</p><p><b>CONCLUSION</b>Changing of PKC activity can regulate and control the expression of TGF beta1, which may play a role in regulating the activation of HSC.</p>
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Animais , Ratos , Linhagem Celular , Células Estreladas do Fígado , Metabolismo , Proteína Quinase C , Metabolismo , Transdução de Sinais , Acetato de Tetradecanoilforbol , Fator de Crescimento Transformador beta1 , MetabolismoRESUMO
<p><b>OBJECTIVE</b>Kupffer cells (KCs) are resident macrophages in the liver. Because the densities and sizes of KCs show a significant overlap with other sinusoidal cells, it is difficult to separate and purify them. We aim to find an improved procedure that could optimize the method for isolation of highly purified rat Kupffer cells.</p><p><b>METHODS</b>In ex vivo rat liver perfusion with pronase E and collagenase IV, density gradient centrifugation by Histodenz and selective attachment of Kupffer cells were used to isolate them. Cell proliferation and morphological characterization were studied under light phase-contrast microscopes; KCs were also studied with transmission electron microscopy and scanning electron microscopy using standard techniques. Immunocytochemistry was used to detect the expression of ED2 CD163 and lysosome associated membrane protein 2 (LAMP2). Phagocytosis of latex-beads by KCs was also studied.</p><p><b>RESULTS</b>The yield rate of KCs was 5 x 10(7) and the KCs viability exceeded 98%. The purity of KCs identified by ED2 was higher than 98%, and over 99% of the collected KCs were LAMP2 positive.</p><p><b>CONCLUSION</b>In the future this simple, stable and effective method of collecting highly purified Kupffer cells is expected to help in further studies.</p>
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Animais , Masculino , Ratos , Técnicas de Cultura de Células , Separação Celular , Métodos , Células Cultivadas , Células de Kupffer , Biologia Celular , Ratos Endogâmicos LewRESUMO
<p><b>OBJECTIVE</b>To investigate the protective effects of Shenfu Injection (SI) on hepatic ischemia-reperfusion injury in rats.</p><p><b>METHODS</b>Partial liver ischemia-reperfusion model under room temperature was established in 60 rats, which were divided into the control group and the treated group randomly and each group was again classified into 3 subgroups with 30 min, 60 min and 90 min hepatic ischemia time rspectively. Rats in the treated group were injected with SI 10 ml/kg every day, while the control group treated with normal saline. Survival rate after 1 week was observed, the serum levels of aspartate aminotransferase (AST), malondialde hyde (MDA), surperoxide dismutase (SOD), tumor nicrosis factor alpha (TNF-a) and endothelin (ET) were detected, and hepatic biopsy was performed with light and electronic microscope.</p><p><b>RESULTS</b>The survival rate in the treated subgroup with 90 min' ischemia after 1 week was 90%, higher than that in the control subgroup significantly (P <0. 05), which was 60% ; and serum levels of AST, MDA, TNF-alpha and ET were lower and SOD was higher significantly (all P <0.05), as well as the degenerative and necrotic degree of hepatocyte and sinusoidal endothelial cells was lighter in the 3 treated subgroups, compared with the control group.</p><p><b>CONCLUSION</b>Shenfu injection can eliminate oxygen free radical during hepatic ishemia-reperfusion so as to has a protective effect and attenuate hepatic ischemia reperfusion injury.</p>
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Animais , Ratos , Aspartato Aminotransferases , Sangue , Medicamentos de Ervas Chinesas , Usos Terapêuticos , Endotelinas , Sangue , Injeções , Fígado , Hepatopatias , Sangue , Tratamento Farmacológico , Malondialdeído , Sangue , Substâncias Protetoras , Usos Terapêuticos , Traumatismo por Reperfusão , Sangue , Tratamento Farmacológico , Superóxido Dismutase , Sangue , Resultado do Tratamento , Fator de Necrose Tumoral alfa , SangueRESUMO
Living related liver transplantation (LRLT) has been developed as a rescue for the ever deteriorating shortage of cadaveric donor liver, especially in Asian countries and areas where the concept of brain death has not widely been accepted. Inclusion criteria for the biologically suitable potential donor of LRLT should be strict and approved by the ethical committee of the hospital before clinical evaluation is taken. Anatomical, physiological and clinical practices have proved that donor mortality is acceptably very low. The early and middle-long term results for the recipient of LRLT is comparable with and even better than those of cadaveric liver transplantation. In mainland China, interest in LRLT is surging with the volume ever increased and donor's safety guaranteed.
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Humanos , Transplante de Fígado , Doadores VivosRESUMO
<p><b>OBJECTIVES</b>To investigate the effect of interlukin-10 (IL-10) on expression and secretion of collagen I, IV in rat's hepatic stellate cells (HSC) of livers injured by CCl4.</p><p><b>METHOD</b>The adenovirus vector encoded IL-10 gene was used to transfect rats with liver injury via the caudal veins. HSC were isolated and purified from the rat livers by collagenase IV perfusion and density gradient centrifugation with Nycodenz. The expression of collagen I, IV mRNA in HSC was detected by semi-quantitative RT-PCR method and the secretion of collagen I, IV in culture serum of HSC by ELISA method. The quantity of collagen was measured in the van Gieson stained histological liver preparations.</p><p><b>RESULTS</b>The expression and secretion of collagen I, IV in the adenovirus vector encoding IL-10 gene group were significantly lower than those in the adenovirus vector without IL-10 gene group and the control group (P < 0.05). The quantity of collagen in the treatment group was lower than that in the control group.</p><p><b>CONCLUSION</b>IL-10 can inhibit collagen I, IV expression and secretion in rat HSC.</p>
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Animais , Masculino , Ratos , Células Cultivadas , Colágeno Tipo I , Genética , Colágeno Tipo IV , Genética , Hepatócitos , Metabolismo , Patologia , Interleucina-10 , Farmacologia , Cirrose Hepática Experimental , Metabolismo , Patologia , Ratos Sprague-DawleyRESUMO
<p><b>OBJECTIVES</b>To screen and clone the genes encoding hepatocellular carcinoma associated tumor antigens.</p><p><b>METHODS</b>A hepatocellular carcinoma cDNA express library was constructed with ZAP vector and analyzed by serological analysis of recombinant cDNA expression library (SEREX) with sera from autologous and allogenous patients. Monoclonalized positive phage clones were converted into pBK-CMV phagemid forms by in vivo excision. The cDNA inserts were determined by restriction endonuclease digestion with EcoR I and Xho I. The cDNA inserts were sequenced and analyzed with bioinformatics. LIMS1 insert was cut from the clone HCL5-70 and constructed into pQE 31 express vector. The recombinant LIMS1 was expressed in M15 and analyzed with SDS-PAGE and Western blot.</p><p><b>RESULTS</b>Fourteen genes were cloned from autologous screening and eleven genes were obtained with allogeneous analysis. One gene, kinectin, was identified in both autologous and allogeneous screening. Eight of the total twenty-four genes were unknown for their functions; the other sixteen genes can be classified into eight groups according to their established or putative function. Recombinant LIMS1 was expressed in M15.</p><p><b>CONCLUSION</b>The identification of hepatocellular carcinoma associated tumor antigens provides potential targets for immunotherapy of hepatocellular carcinoma patients and will help in the understanding of the carcinogenesis of hepatocellular carcinoma.</p>
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Humanos , Antígenos de Neoplasias , Genética , Alergia e Imunologia , Carcinoma Hepatocelular , Genética , Alergia e Imunologia , DNA Complementar , Genética , Regulação Neoplásica da Expressão Gênica , Terapia Genética , Neoplasias Hepáticas , Genética , Alergia e ImunologiaRESUMO
<p><b>OBJECTIVE</b>To study the risk factors of renal failure in the early post-liver transplantation period.</p><p><b>METHODS</b>92 consecutive liver transplantation cases were reviewed and a multi-factor analysis of presumed risk factors of early post-transplantation period renal failure was conducted. The factors analyzed were total bilirubin level, prothrombin activity, onset of structural renal disease, onset of gastrointestinal hemorrhage, whether the patient underwent large-volume paracentesis, or underwent plasmapheresis therapy, needed renal replacement therapy, the operation method used, the bleeding volume during operation and the immunosuppressive agents used.</p><p><b>RESULTS</b>Of the 92 patients, 29 (31.5%) developed acute renal failure (ARF) in the early postoperative period. Multi-factor analysis revealed a high pre-transplantation serum creatinine level and low prothrombin activity as risk factors for development of ARF.</p><p><b>CONCLUSION</b>ARF is a frequent medical complication after liver transplantation. A high pre-transplantation serum creatinine level and low prothrombin activity are risk factors of its development.</p>
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Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Injúria Renal Aguda , Epidemiologia , China , Epidemiologia , Creatinina , Sangue , Cirrose Hepática , Cirurgia Geral , Transplante de Fígado , Protrombina , Metabolismo , Fatores de RiscoRESUMO
<p><b>OBJECTIVE</b>To construct dendritomas by fusion of human dendritic cells with HLE cells, a human hepatocellular carcinoma cell line.</p><p><b>METHODS</b>HLE cells were cultured in RPMI 1640 with 15% FCS. Human dendritic cells (DCs) were obtained from peripheral blood monocytes cultured in the presence of GM-CSF and IL-4 for 7 days, matured with TNF-alpha and PGE(2) for 2 days. The DCs and HLE cells were labeled with green fluorescence dye PKH67-GL and red fluorescence dye PKH26-GL, respectively, and fused in 50% polyethylene glycol (PEG) + 10% dimethyl sulfoxide (DMSO) to generate dendritomas for rapid fluorescence-activated cell sorting (FACS).</p><p><b>RESULTS</b>Dendritomas with dual red-green fluorescence were constructed successfully, and FACS analysis showed the effective fusion rate was 16.8%.</p><p><b>CONCLUSION</b>With fluorescence dyes PKH67-GL and PKH26-GL as fusion markers, dendritomas for rapid fluorescence-activated cell sorting are constructed, which may throw new light on immunotherapy of hepatocellular carcinoma.</p>
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Humanos , Vacinas Anticâncer , Carcinoma Hepatocelular , Patologia , Fusão Celular , Métodos , Linhagem Celular Tumoral , Células Cultivadas , Células Dendríticas , Biologia Celular , Células Híbridas , Neoplasias Hepáticas , PatologiaRESUMO
<p><b>OBJECTIVE</b>To investigate the variations type of hepatic artery and discuss the method of how to protect hepatic artery from injury during the quick harvest of the donor liver.</p><p><b>METHODS</b>A retrospective review of the variations of hepatic arteries of the donor livers and the course of excision and reconstruction of 200 donor livers was performed, and the aberrance and reconstruction method of hepatic arteries were summarized.</p><p><b>RESULTS</b>37 out of 200 hepatic arteries varied and 2 patients suffered biliary complications because of improper preservation of aberrant hepatic arteries.</p><p><b>CONCLUSIONS</b>Most aberrant liver arteries come from superior mesenteric artery or left gastric artery. Proper quick harvest of multiple organs is the basis of the integrity of hepatic arteries, and all the aberrance must be reconstructed.</p>
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Feminino , Humanos , Masculino , Artéria Hepática , Cirurgia Geral , Transplante de Rim , Transplante de Fígado , Métodos , Estudos Retrospectivos , Coleta de Tecidos e Órgãos , Métodos , Transplante HomólogoRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of interleukin-10 (IL-10) on the expression of transforming growth factor-beta(1) (TGFbeta(1)) and platelet-derived growth factor (PDGF) in hepatic stellate cells (HSC) during liver injury.</p><p><b>METHODS</b>The adenovirus vector (the titer was 1 x 10(7) efu/ml) encoded IL-10 gene was used to transfect the rat via the vein of caudal. At the same time, CCl(4) was injected into rat by a hypodermic injection. These processes went on twice a week. After eight weeks, the liver were perfused with collagenase IV and purified by density gradient centrifugation with Nycodenz for separate HSC. The level of IL-10 was measured by ELISA method; The expression of PDGF and TGFbeta(1) in HSC was detected by semi-quantitative RT-PCR and Western-blot methods.</p><p><b>RESULTS</b>The level of IL-10 in therapy group (adenovirus vector encoding IL-10 gene group) was higher than that in non-therapy group (adenovirus vector without IL-10 gene and PBS group); The expression of TGFbeta(1) mRNA, TGFbeta(1) protein and PDGF mRNA, PDGF protein in therapy group were significantly lower than that in non-therapy group (P < 0.05).</p><p><b>CONCLUSION</b>Downregulating the TGFbeta(1) and PDGF expression could be the passageway by which IL-10 alleviate the degree of proliferation and activation in hepatic stellate cells.</p>
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Animais , Masculino , Ratos , Regulação para Baixo , Terapia Genética , Hepatócitos , Fisiologia , Interleucina-10 , Farmacologia , Cirrose Hepática Experimental , Metabolismo , Patologia , Terapêutica , Fator de Crescimento Derivado de Plaquetas , RNA Mensageiro , Ratos Sprague-Dawley , Células Estromais , Fisiologia , Transfecção , Fator de Crescimento Transformador beta , Fator de Crescimento Transformador beta1RESUMO
<p><b>OBJECTIVE</b>To evaluate the effects of portaazygous disconnection (PAD), portacaval shunt (PCS) and distal splenocaval shunt (DSCS) on the portosytemic shunting (PSS), hepatic function (HF), hepatic mitochondrial respiratory function (HMRF), oral glucose tolerance test (OGTT) and arterial ketone body ratio (KBR) in order to provide a sound basis for selecting suitable operations for patients.</p><p><b>METHODS</b>Using a cirrhotic portal hypertensive model induced by CCl4/ethanol in Wistar rats, the PSS, HF, HMRF, OGTT and KBR were determined three weeks after PCS, DSCS and PAD.</p><p><b>RESULTS</b>It was revealed that: (1) In the cirrhotic portal hypertension rats, the PSS increased significantly, HMRF and hepatic reserve function (HRF) decreased significantly when compared with the control rats. (2) At the time of first postoperative week, the mean blood glucose value in the 120-minute OGTT in each PAD, PCS and DSCS groups had significant differences compared with the cirrhotic control group. But during the second and third postoperative weeks, the mean blood glucose values in the 120-minute OGTT in both PAD and DSCS groups had no significant differences compared with the cirrhotic control group except for the PCS group. The values of KBR in the three operative groups decreased significantly compared with the cirrhotic control group during the two postoperative weeks. In the third postoperative week, only the values of KBR in the PCS group had a significant difference compared with the cirrhotic control group. (3) After PCS, the PSS was further increased; HF and HMRF were significantly decreased. Little improvement was found in the third postoperative week. (4) After DSCS and PAD, the above mentioned indices were less influenced, and they were restored more quickly than those in the PCS group.</p><p><b>CONCLUSION</b>We found that PAD and DSCS are more desirable than PCS.</p>
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Animais , Ratos , Hipertensão Portal , Cirurgia Geral , Cirrose Hepática Experimental , Cirurgia Geral , Derivação Portocava Cirúrgica , Derivação Portossistêmica Cirúrgica , Métodos , Ratos WistarRESUMO
<p><b>OBJECTIVE</b>To investigate the regulatory effect of interleukin-10 (IL10) on the activation of hepatic stellate cells (HSC) through platelet derived growth factor (PDGF) and mitogen-activated protein kinase (MAPK) pathways.</p><p><b>METHODS</b>HSC were divided randomly into 4 groups. Group 1 served as a control. HSC were incubated with 1 ng/ml, 5 ng/ml, and 25 ng/ml IL-10 in groups 2, 3 and 4. RT-PCR and western blot were used to detect the expression of PDGF and MAPK protein ERK and p38 and alpha-SMA.</p><p><b>RESULTS</b>Compared with the control group, expressions of ERK, p38 and alpha-SMA of groups 2, 3 and 4 were significantly lower (F values were 240.47, 21.39, 28.86 respectively. IL-10 inhibited PDGF and MAPK protein ERK and p38 and alpha-SMA expression in a dose-dependent way.</p><p><b>CONCLUSION</b>IL-10 inhibits activation of HSC through the PDGF/MAPK pathway.</p>
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Animais , Ratos , Linhagem Celular , Proliferação de Células , Hepatócitos , Biologia Celular , Interleucina-10 , Farmacologia , Proteínas Quinases Ativadas por Mitógeno , Fator de Crescimento Derivado de Plaquetas , Transdução de SinaisRESUMO
<p><b>OBJECTIVE</b>To investigate the expression of MAGE-B genes in hepatocellular carcinoma (HCC) in order to find new targets for immunotherapy.</p><p><b>METHODS</b>The expression of MAGE-B1, B2, A1 and A3 mRNA was detected using RT-PCR in HCC tissues and the corresponding adjacent non-HCC tissues from 47 HCC patients, 30 samples of cirrhosis and normal liver tissues. Four samples selected randomly from MAGE-B1 or B2 with positive RT-PCR results were sequenced to confirm the results of RT-PCR. The relationship between the expression of MAGE-B and some clinicopathological parameters was analyzed.</p><p><b>RESULTS</b>MAGE-B1 mRNA and MAGE-B2 mRNA were detected in 44.7% (21/47) and 61.7% (29/47) of HCC samples, respectively, while neither MAGE-B1 nor MAGE-B2 could be detected in the corresponding adjacent non-HCC liver tissues. In addition, none of 30 samples of cirrhosis and normal liver tissues was shown to express both MAGE-B genes. The DNA sequence confirmed that the RT-PCR products were truly target cDNA. The frequency of the expression of MAGE-A1 and A3 was 74.5% (35/47) and 44.7% (21/47), respectively. There was significant correlation between the expression of MAGE-B and MAGE-A (P < 0.05). However, the positive expression of MAGE-B was observed in 5 out of 12 HCC tissues without expression of MAGE-A1 and/or A3. When all four MAGE genes were examined, the positive rate of expression of one, two, three and four genes was 83.0% (39/47), 55.3% (26/47), 48.9% (23/47), and 38.3% (18/47) of 47 HCC tissues, respectively. No correlation was found between the expression of MAGE-B and clinical parameters such as age, sex, tumor size, degree of tumor differentiation, serum alpha-fetoprotein level and hepatitis B virus or hepatitis C virus infection (P > 0.05).</p><p><b>CONCLUSION</b>MAGE-B genes are expressed with relatively high frequency and specificity in HCC. Most HCC patients with positive expression of at least one member of MAGE-B or MAGE-A gene family are adequate candidates to receive specific immunotherapy. Frequent co-expression of multiple members of MAGE-B and MAGE-A subfamilies provides the possibility of using polyvalent vaccines to achieve more effective immunotherapeutic results.</p>
Assuntos
Humanos , Antígenos de Neoplasias , Genética , Carcinoma Hepatocelular , Genética , Expressão Gênica , Neoplasias Hepáticas , Genética , Antígenos Específicos de Melanoma , Proteínas de Neoplasias , Genética , Prognóstico , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
<p><b>OBJECTIVE</b>To study the therapeutic effects to block the TGF-beta1 (transforming growth factor beta1) signal transduction by antisense Smad4 gene on experimental fibrotic liver.</p><p><b>METHODS</b>Using the rat model of liver fibrosis induced by Carbon Tetrachloride (CCl4)/ethanol, we transfected antisense Smad4 gene mediated by adenovirus via portal vein infusion into the liver, and observed the expression of Smad4 by Retro-Polymerase Chain Reaction (RT-PCR) and Western Blot. We also investigated the pathologic features and collagen expression.</p><p><b>RESULTS</b>In the non-therapeutic cirrhotic liver, the expression of Smad4 mRNA was significantly increased than normal liver, and so was the collagen I. After antisense Smad4 gene being transfected, the expression of Smad4 mRNA and that of collagen I in the therapeutic liver was significantly decreased, compared with the non-therapeutic cirrhotic liver. The fibrous degree of therapeutic liver was also reduced compared with the non-therapeutic fibrous liver.</p><p><b>CONCLUSION</b>These results indicate that because antisense Smad4 gene could block TGF-beta1 signal transduction by reducing the expression of Smad4, so it could inhibit the production of extracellular matrix (ECM) and improve hepatic fibrosis.</p>