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Clinical Analysis of Hepatolithiasis with HepatoCholangiocarcinoma

目的:探讨肝胆管结石合并胆管癌的临床表现特征和诊治经验。方法:分析了自2001年1月到2009年8月我院收治的30例肝胆管结石合并胆管癌患者的临床资料。结果:肝胆管结石合并胆管癌术前确诊10例,术中病理检查确诊14例,术后病理检查发现6例。左肝胆管癌16例,占533%,右肝胆管癌7例占233%,肝门部胆管癌5例占167%,其余2例为左右肝都有。肿瘤根治性切除术17例(567%),姑息性手术9例(30%),单纯活检4例(133%)。在30例随访资料中,根治性切除术者平均存活时间26个月。存活1年以上10例,2年以上5例,3年以上2例;姑息治疗者术后平均生存9个月;其余的≤4个月。结论:胆石症状易反复发作,胆道病史较长大于10年且术前CA199gt;2500 ng/mL的患者应高度怀疑合并有胆管癌。

Expression and Significance of Livin in Human Cholangiocarcinoma Tissues and Cell Line

目的:探讨Livin基因在人胆管癌组织及胆管癌细胞系中的表达情况及其与胆管癌发 生发展之间的关系。方法:采用免疫组织化学技术（SP法）检测Livin基因蛋白在45例人胆管 癌标本及及40例癌旁胆管组织、20例正常胆管组织标本中的表达；同时采用RTPCR法及SP 法检测了Livin基因mRNA和蛋白在人胆管癌细胞系QBC939及非肿瘤细胞系HT1080中表达。 结果:Livin在胆管癌组织中表达阳性率为57.8%，而癌旁胆管组织、非癌胆管组织中未能检 测到Livin表达。Livin表达与性别、年龄、肿瘤大小及肿瘤分化程度无关。在有淋巴结转组 中，Livin阳性表达率（70.4%）明显高于无淋巴结转移组（38.9%）。在人胆管癌细胞QBC939 中，Livin mRNA及蛋白均特异性表达，而非肿瘤细胞系HT1080未见Livin表达。结论:Livin 基因在人类胆管癌组织和细胞系中选择性高表达，其可能与胆管癌发生、发展及预后密切相 关。

Treatment Experience of Type Ⅳ Hilar Cholangiocarcinoma

Objective　To explore primary surgical treatment experience of typeⅣ hilar cholangiocarcinoma. Methods　From April 2008 to April 2011，20 patients with type Ⅳ hilar cholangiocarcinoma were enrolled into the same surgical group in Department of Hepatobiliary and Pancreatic Surgery of West China Hospital of Sichuan University.The intra- and post-operative results were analyzed.Results　The total resection rate was 75%，which was consisted of 10 cases of radical excision and 5 cases of non-radical excision.Seven patients received left hepatic trisegmentectomy and caudate lobe resection including anterior and posterior right hepatic duct reconstruction，hepatojejunostomy，and Roux-en-Y jejunojejunostomy.Six patients received enlarged left hepatic trisegmentectomy and caudate lobe resection including left intrahepatic and extrahepatic duct reconstruction，hepatojejunostomy，and Roux-en-Y jejunojejunostomy. Two patients received quadrate lobe resection including two cholangioenterostomies after anterior and posterior right hepatic duct reconstruction，and left intrahepatic and extrahepatic duct reconstruction.After percutaneous transhepatic cholangial drainage （PTCD） and portal vein embolization （PVE），two patients with total bilirubins ＞400 mmol/L received radical excision and non-radical excision，respectively.Three patients only received PTCD during operation due to wide liver and distant metastasis，and two patients received T tube drainage during operation and postoperative PTCD due to left and right portal vein involvement. All 15 patients who received lesion resection survived more than one year， whereas another five patients whose lesions can not been resec ted only survived from 3 to 6 months with the mean of 4.2 months.No death occurred during the perioperative period. Conclusions　For patients with type Ⅳ hilar cholangiocarcinoma， preoperative evaluation and tumor resection shall conducted so as to relieve obstruction of biliary tract，otherwise PTCD and PVE prior to the final lesion resection shall be performed.

Expression of Caspase-3 in Targeted Therapy with Magnetic Nanoparticle in Cholangiocarcinoma Xenograft of Nude Mice

Objective　To investigate the expression of caspase-3 in xenograft that was treated with targeted therapy with magnetic nanoparticles in nude mice. Methods　QBC939 cell lines were injected into nude mice subcutaneously to establish the model of human cholangiocarcinoma xenograft. After two weeks of tumor inoculation, the animal models were divided randomly into 4 groups: group A received placebo (sodium chloride), group B were treated with magnetic nanoparticles (250 mg/kg), group C were treated with magnetic nanoparticles (150 mg /kg) combined with inner-stent, group D with magnetic nanoparticles (250 mg /kg) combined with inner-stent (the inner-stent was used to generate the magnetic targeting effect). The 21th day after treatment, expression of caspase-3 in tumor cells of each groups were measured with histochemical method and RT-PCR. Results　The quantity of caspase-3 in tumor cells that were treated with magnetic nanoparticles (250 mg/kg) combined with inner-stent was the most (P＜0.05), and the quantity of caspase-3 in cells of group C was significantly more than that of the other two groups (P＜0.05). While the quantity of caspase-3 in group B was more than that of the control group(P＜0.05). Conclusion　The use of magnetic nanoparticles combined with inner-stent may increase the expression of caspase-3, and the expression is dose-dependent with magnetic nanoparticles.

In Vivo Antitumor Activity of HSV tk/CD Combinative Gene Toward Human Cholangiocarcinoma Cells(QBC939)

【Abstract】Objective To study the antitumor activity of HSVtk/CD combinative gene toward human cholangiocarcinoma in vivo. Methods Nude mouse models with transplanted subcutaneous cholangiocarcinoma were constructed and divided into 4 groups randomly, each group had 8 mice. Adenovirus solution free from suicide gene was injected in subcutaneous tumors of each mouse of control group. Adenovirus solution containing cytosine deaminase (CD), thymidine kinase (tk) and HSVtk/CD fusional gene were injected into single suicide gene either HSVtk or CD was transinfected into the tumor cells by injecting viras into subcutaneous tumor of mice of CD gene,tk gene and fused CD and tk gene group respectively. 24 hours after the injection, 5fluorocytosine (5FC) and ganciclovir (GCV) were injected introabdominally in each mouse. Growth of the tumors were monitored.Results Tumor growth of the genetransfection groups was suppressed in different degrees. Compared with the control group, the suppressing rates of the genetransfection groups were 41.2%, 55.7% and 70.0% respectively (P＜0.05). Histological examination showed good tumor growth in the control group, and tumor necrosis in the other 3 groups, particularly obvious in the group transfected with pAd(HSVtk/CD).Conclusion Combinative gene system has a b antitumor effect on cholangiocarcinoma in vivo. But it’s not powerful enough to eliminate tumor thoroughly because of insufficient “Bystander effect”.

Expression of Beta-Catenin in Hilar Cholangiocarcinoma and Relevance to the Expression of c-myc Gene

【Abstract】ObjectiveTo investigate whether abnormal expression of β-catenin and high expression of c-myc have played a possible role in hilar cholangiocarcinoma carcinogenesis.MethodsBy using immunohitochemical staining (SP method), the authors detected the expression of β-catenin and c-myc in 42 paraffin-embedded samples of hilar cholangiocarcinoma and 10 benign bile duct disease tissue, and then analyzed the relationship of them with clinical data. Resultsβ-catenin was normally expressed in 10 benign bile duct disease tissue, while expression of c-myc was negtive. In hilar cholangiocarcinoma tissue, the positive expression rate of β-catenin （71.4%） was significantly correlated to the lymphoid node metastasis of hilar cholangiocarcinoma (χ2=4.75，P＜0.05),but was not statistically correlated to the tumor size，the extent of differentiation and infiltration (χ2=3.35，3.45，4.32，Pgt;0.05); the expression rate of c-myc (76.2%) was correlated with the extent of differentiation(χ2=4.87, P＜0.05),but not with the size, infiltration, lymphoid metastasis（χ2= 3.47，4.12，2.76， Pgt;0.05). The abnormal expression of β-catenin had relevance to the high expression of c-myc with hilar cholangiocarcinoma （r=0.324,P＜0.01）. ConclusionThe expression of beta-catenin and c-myc is significantly altered in hilar cholangiocarcinoma, and correlate with biological features of cholangiocarcinoma.The abnormal expression of beta-catenin is one of the mechanisms for the spread of hilar cholangiocarcinoma.

Analysis of Factors Influencing Prognosis of Patients with Bile Duct Carcinoma after Resection

【Abstract】ObjectiveTo analyze the factors influencing the prognosis of patients with bile duct carcinoma after resection. MethodsThe clinical data of 120 patients with bile duct carcinoma receiving resection in our hospital from 1980 to 2004 were collected retrospectively and clinicopathologic factors that might influence survival were analysed. A multiple factor analysis was performed through Cox proportional hazard model. ResultsThe overall 1year, 3year and 5year survival rates were 71.7%, 32.5% and 19.2% respectively. The single factor analysis showed that the major significant factors influencing survival of these patients were histological type of the lesions, lymph node metastasis, pancreatic infiltration, duodenal infiltration, resected surgical margin, perineural infiltration, peripheral vascular infiltration and depth of tumor infiltration (P＜0.05). Lymph node metastasis, pancreatic infiltration and perineural infiltration were found to be the the statistically significant factors influencing survival by multiple factor analysis through the Cox model. ConclusionThe most important prognostic factors for bile duct carcinoma after resection were lymph node metastasis, pancreatic infiltration and perineural infiltration.

Effect of Transfection with Antisense DNMT3b Gene Eukaryotic Expression Vector on Expression of DNMT3b Gene in Human Cholangiocarcinoma Cell Line

【Abstract】ObjectiveTo study the effect of transfection with antisense DNMT3b gene eukaryotic expression vector on the expression of DNMT3b gene in human cholangiocarcinoma cell line QBC-939. MethodsThe constructed antisense DNMT3b gene eukaryotic expression vector was transfected into the human cholangiocarcinoma cell line QBC-939 by using lipofectamine transfection reagents, and positive cell clones were obtained by using G418 selection after transfection. Whether the constructed recombinant vector was transfected into QBC-939 cells successfully was confirmed by amplifying the exogenous neoR gene with PCR method. The expression of DNMT3b gene mRNA and protein were detected by semi-quantitative RT-PCR and FCM methods respectively. ResultsFollowing the transfection of antisense DNMT3b gene eukaryotic expression vector, the mRNA level of DNMT3b gene in QBC-939 cells of human cholangiocarcinoma decreased from 0.956±0.053 to 0.209±0.023, and the protein level of DNMT3b gene also decreased from (75.38±3.22)% to (29.87±3.46)%. There were very significant differences on the expression levels of DNMT3b gene between non-tranfections group and the antisense DNMT3b gene eukaryotic expression vector transfection group (P＜0.01). ConclusionTransfection with antisense DNMT3b gene eukaryotic expression vector significantly reduces the expression level of DNMT3b gene in human cholangiocarcinoma cell line QBC-939, and this study may provide a valid tool and method to investigate the function of DNMT3b gene and its role in cholangiocarcinoma.

Expression of Vascular Endothelial Growth Factor and c-myc in Cholangiocarcinoma and Its Significance

Cholangiocarcinoma; Vascular endothelial growth factor; c-myc gene

Perspectives on the Progress of Surgical Treatment of Hilar Bile Duct Cancer