不同组织微环境中人胃癌异种移植瘤生物学行为及明胶酶的表达差异

目的:探讨不同组织微环境中明胶酶在异种移植瘤中的表达差异及其对移植瘤生物学行为的影响.方法:?应用原位杂交方法分别检测皮下移植瘤及腹腔移植瘤中MMP-2?mRNA,?MMP-9?mRNA的表达情况,?并应用常规HE染色观察不同移植瘤的生物学行为表现.结果:?人胃癌裸小鼠皮下异种移植瘤呈膨胀性生长,?侵袭性不明显;?腹腔内移植瘤呈侵袭性生长;?MMP-2及MMP-9?mRNA在皮下移植瘤细胞及间质表达阴性,?在腹腔移植瘤为阳性表达,?且表达呈部位特异性,?即癌巢边缘部位的癌细胞胞质和/或包膜阳性表达.结论:?相同来源的瘤组织在不同的微环境中,?生长方式明显不同,?组织环境对肿瘤侵袭表型有很大影响;?明胶酶的表达与肿瘤生长方式及侵袭性有密切联系.     

晚期肝细胞癌靶向治疗及个性化治疗的现状

随着精准医疗理念的普及,在晚期肝细胞癌系统性治疗领域,新技术驱动的个性化治疗逐渐流行,晚期肝细胞癌靶向治疗也迎来了新的发展契机。本文就晚期肝细胞癌靶向治疗的现状与过去、新技术驱动的个性化治疗的发展与未来进行综述,着重介绍以sorafenib为代表的分子靶向药的发展历程,以及二代测序、人源肿瘤组织异种移植模型等新技术在个性化精准治疗中的应用。     

胰腺肿瘤的基础转化研究模型

在多种癌症治疗取得突破的今天,胰腺癌治疗药物的研发仍举步维艰,缺少能反映肿瘤细胞生物学特性和模拟其复杂微环境的模型是导致从基础到临床转化研发失败的重要原因。随着类器官、人源肿瘤异种移植和人源化小鼠等技术的出现,使个体化精准研究肿瘤异质性、高度模拟包含间质细胞和免疫细胞的肿瘤微环境成为可能。利用这些新技术开展胰腺癌的基础和转化研究可以弥补既往研究手段的不足,有可能建立适用于胰腺癌研究的精准模型,推动胰腺癌治疗药物的研发,改善胰腺癌的疗效。     

三种来源细胞因子诱导的杀伤细胞对人食管癌裸鼠原位移植肿瘤的转移抑制作用

目的探讨三种来源的细胞因子诱导的杀伤(CIK)细胞对人食管癌裸鼠原位移植肿瘤的转移抑制作用。方法建立人食管癌裸鼠原位移植模型。取32只荷瘤裸鼠平分为四组:食管癌患者自体外周血CIK细胞组、健康人外周血CIK细胞组、脐血CIK细胞组及生理盐水对照组。给予20 d对应的注射治疗,停药72 h后处死裸鼠,称量各组原位移植瘤瘤重,计算抑瘤率;观察肿瘤转移情况;免疫组织化学法检测基质金属蛋白酶-9(MMP-9)的表达情况。结果与对照组相比,CIK细胞治疗组的肿瘤生长及转移明显受到抑制(P<0.05)。且CIK细胞可抑制肿瘤组织中MMP-9的表达。结论三种来源CIK细胞对人食管癌裸鼠原位移植肿瘤的生长及转移有明显抑制作用,且脐血来源的CIK细胞抗瘤作用最强,其抗转移作用可能与肿瘤组织中MMP-9的表达下降有关。     

异种猪心瓣膜的现况:5年评价

异种心脏瓣膜移植后发生血栓栓塞的危险性很小,许多病人可不需抗凝治疗。这种异种移植组织必须能够储存,又能摒除免疫反应,且能消毒灭菌。将异种瓣膜缝置在支架上,使之可以采用和移植机械人造瓣膜相似的操作。然而,移植后组织的形态学变化,限制了瓣膜的耐久性,这是一个需要长期注意的问题。     

两种不同方法建立脑胶质瘤裸鼠移植模型及组织学特征分析

采用C6型鼠胶质瘤细胞悬液接种至裸鼠皮下,同时用瘤组织块异体移植至裸鼠皮下,建立脑胶质瘤细胞裸鼠移植动物模型,分别观察裸鼠皮下移植瘤长出时间,并分析肿瘤的病理组织学及免疫组织化学特征.结果 两种方法所建肿瘤模型其成功率均为100%,均符合胶质瘤细胞的形态学特征.采用瘤组织块异体移植法建立的肿瘤模型,所需时间均明显短于细胞接种法,裸鼠肿瘤组织块生长速度基本相同.认为利用肿瘤组织块移植的方法可快速建立裸鼠皮下移植瘤模型,并能作为研究胶质瘤发病机制、生物学特性以及基因治疗的可靠动物模型.     

人舌癌移植瘤模型中血管内皮细胞来源的实验研究

目的 观察人舌癌裸鼠皮下移植瘤模型中肿瘤新生血管内皮细胞的来源.方法 通过皮下移植人舌癌Tca8113-Ml细胞建立人舌癌裸鼠移植肿瘤模型,并于肿瘤生长至直径1cm时切除肿瘤,对移植瘤进行病理组织学观察,利用免疫组织化学方法检测移植瘤内新生血管内皮细胞的来源,抗体为鼠抗人单克隆抗体CD34和大鼠抗小鼠单克隆抗体CD34;并进行微血管密度(microvessel density,MVD)计数及血管内皮生长因子(vascular endothelial growth factor,VEGF)表达检测.结果 右腋窝皮下接种人舌癌Tca8113-Ml细胞2周后,6只裸鼠的肿瘤直径都达到1 cm 以上,切片HE染色可见肿瘤组织病理形态为鳞状上皮细胞癌,MVD平均值为10.72±2.12,其中肿瘤新生血管内皮细胞大鼠抗小鼠CD34免疫组化结果是阳性,而鼠抗人CD34阴性.VEGF在6例移植瘤标本中均阳性表达, 平均阳性率为(67±5.6)%.结论 人舌癌移植瘤模型中血管内皮细胞主要来源于宿主裸鼠,并且可能与肿瘤细胞分泌的VFGE诱导有关.     

仓鼠-大鼠异种小肠移植模型的建立及排斥反应初步观察

同种供器官的短缺使异种移植逐渐受到重视［１］，然而对异种移植的免疫反应特点仍未完全了解．我们在建立稳定的同种小肠移植模型基础上通过对仓鼠消化道解剖特点的了解改进建立了仓鼠一大鼠小肠移植模型并初步观察了其排斥反应及病理特点．１临床资料实验动物取♂叙利亚...     

人胰腺癌鸡胚模型的建立及其肿瘤生物学观察

目的建立人胰腺癌鸡胚移植模型,为临床胰腺癌研究提供一种简便实用的工具.方法将人胰腺癌细胞系SW1990细胞接种到鸡胚绒毛尿囊膜(CAM)上,动态观察移植瘤的形态及肿瘤生物学特性,分析影响移植瘤成活的因素,并对瘤组织行组织学检查.结果成功建立了人胰腺癌鸡胚移植模型,移植瘤组织学结构与人胰腺癌相似;接种癌细胞数量影响接种成瘤率,接种癌细胞数量越大,成瘤率越高;移植瘤可诱发血管生成并向肿瘤呈放射状集中.结论将SW1990细胞接种到鸡胚上建立鸡胚移植胰腺癌是可行的,本模型可动态观察胰腺癌生长,模拟其在体内生长情况,为胰腺癌研究提供一种简便实用的工具.     

仓鼠到大鼠原位肝移植38例

近年来,由于同种肝移植的巨大进步,供肝来源不足的矛盾日益尖锐,人们的兴趣重新转向异种移植,兴起了异种移植研究的热潮［１］．仓鼠到大鼠肝脏移植为协调性异种移植［２］,因其排斥反应稳定,来源丰富,价格便宜,是开展异种肝移植基础研究较好模型之一,但手术难度较大,限制了使用．本实验在参考Ｈａｒｉｈａｒａｅｔａｌ、Ｋａｍａｄａｅｔａｌ［３,４］所介绍的方法的基础上,加以改进,简化了手术操作,缩短了手术时间,并探索出预防各种并发症的有效措施,提高了手术成功率,为广泛开展异种肝移植基础研究提供了良好稳定的实验…     

N-乙酰基转移酶基因多态性与肝癌易感性的关系

目的探讨N-乙酰基转移酶(NAT2)基因多态性与肝癌易感性的关系。方法应用自动实时荧光Light-Cycler技术,分析78例肝癌患者和112例健康志愿者NAT24个位点的基因多态性,比较肝癌患者与对照组间频率差异。结果 肝癌吸烟组NAT2慢乙酰化基因型频率(37.5％)与对照吸烟组(17.9％)比较差异有显著性(X2=4.67,P<0.05),并使患肝癌的危险度提高了2.76倍;肝癌非吸烟组NAT2慢乙酰化基因型频率(26.3％)与对照非吸烟组(16.1％)比较差异无显著性(X2=1.47,P>0.05)。结论携带NAT2慢乙酰化基因型的吸烟者可能是肝癌的高危人群。     

Hepatocellular carcinoma arising in the absence of cirrhosis in genetic haemochromatosis: three case reports and review of literature.

Genetic haemochromatosis constitutes a high risk factor for the development of hepatocellular carcinoma. It is widely accepted that venesection prevents the evolution of cirrhosis in haemochromatosis and indirectly protects against the development of hepatocellular carcinoma. Clinical, pathological and radiological data are presented on three patients who did not conform to the 'siderosis-cirrhosis-carcinoma' sequence and in whom prompt and adequate iron depletion did not prevent the development of cancer. This is the first report of hepatocellular carcinoma intervening in non-cirrhotic liver in two siblings with genetic haemochromatosis. The current literature on the subject is reviewed. The direct oncogenic role of iron remains to be elucidated. Hepatocellular carcinoma should be considered as a differential diagnosis in patients with non-cirrhotic genetic haemochromatosis who present with clinical deterioration during the course of an otherwise uneventful venesection programme.     

Hepatocellular carcinoma: molecular and genomic guideline for the clinician

Understanding of the genetic changes and molecular signaling pathways that are active in hepatocellular carcinoma has improved substantially over the last decade. As more information becomes available, it is clear that the prognostication of hepatocellular carcinoma will soon include molecular and genomic "fingerprints" that are unique to each cancer, which will allow more personalized treatment plans for patients as more targeted therapies become available. This article discusses the molecular and genomic changes that are important in hepatocellular carcinoma in order for clinicians to understand the current and forthcoming treatment options for patients with liver cancer.     

Histocompatibility antigens in patients with hepatocellular carcinoma and their relationship to chronic hepatitis B virus infection in these patients.

Although hepatocellular carcinoma is probably caused by one or more environmental carcinogens, a genetically determined susceptibility to the development of the tumor has not been excluded. In looking for such a predisposition, we have compared the histocompatibility antigens (HLA) of 102 southern African blacks with histologically proved HCC with those of 208 healthy blacks. The standard two-stage lymphocyte microcytotoxicity method was used to test for 40 antigens: 17 in the A locus, 20 in the B locus, and 3 in the C locus. None of the HLA antigens had a frequency that was significantly different in the patients and the controls. A close association undoubtedly exists between chronic hepatitis B virus infection and hepatocellular carcinoma. If this virus is proved to be oncogenic with respect to hepatocellular carcinoma, a genetic predisposition to the hepatitis B virus carrier state may have an indirect bearing on the etiology of the tumor. Sera from the hepatocellular carcinoma patients were therefore tested for hepatitis B virus markers (HBV surface antigen and antibody against HBV core antigen), and these were related to the patients' histocompatibility antigens. None of the HLA antigen frequencies was significantly different in the surface antigen-positive and the surface antigen-negative patients. As 88% of the patients were anticore positive, no meaningful correlation could be carried out with this marker. Analysis of histocompatibility antigens thus failed to show evidence of a genetic predisposition either to hepatocellular carcinoma or to chronic hepatitis B surface antigenemia in patients with this tumor.     

Diabetes mellitus worsens the recurrence rate after potentially curative therapy in patients with hepatocellular carcinoma associated with nonviral hepatitis

Background and Aim: The aim of this retrospective study was to examine the relationship between diabetes mellitus and recurrence of hepatocellular carcinoma after potentially curative therapy for hepatocellular carcinoma with nonviral hepatitis. Methods: We studied 40 consecutive hepatocellular carcinoma patients who were diagnosed between 1980 and 2006 with hepatocellular carcinoma associated with non‐B, non‐C hepatitis, and later underwent surgical resection or radiofrequency ablation. Results: Twenty‐two out of the 40 patients developed hepatocellular carcinoma recurrence within a median of 3.7 years. In the 18 patients with diabetes mellitus, the cumulative rates of hepatocellular carcinoma recurrence were 22.2% at the first year, 55.6% at the second year, 61.1% at the third year, 61.1% at the fourth year, and 80.6% at the fifth year. The cumulative rates of hepatocellular carcinoma recurrence in 22 nondiabetic patients were 24.6% at the first year, 24.6% at the second year, 31.5% at the third year, 31.5% at the fourth year, and 31.5% at the fifth year. The hepatocellular carcinoma recurrence rate was significantly higher in diabetic patients than in nondiabetics (P = 0.026). The multivariate Cox proportional model identified old age and diabetes as the only significant predictors for recurrence. The hazard ratio of hepatocellular carcinoma recurrence in diabetic patients was 4.61 (P = 0.007). There was no significant difference in overall survival rate between diabetic and nondiabetic patients (P = 0.392). Conclusion: Diabetes is a significant predictor of tumor recurrence after potentially curative therapy for hepatocellular carcinoma in patients with nonviral hepatitis.     

Prognostic factors and survival in patients with hereditary hemochromatosis and cirrhosis.

OBJECTIVES: The survival of treated, noncirrhotic patients with hereditary hemochromatosis is similar to that of the general population. Less is known about the outcome of cirrhotic hereditary hemochromatosis patients. The present study evaluated the survival of patients with hereditary hemochromatosis and cirrhosis. METHODS: From an established hereditary hemochromatosis database, all cirrhotic patients diagnosed from January 1972 to August 2004 were identified. Factors associated with survival were determined using univariate and multivariate regression. Survival differences were assessed using the Kaplan-Meier life table method. RESULTS: Ninety-five patients were identified. Sixty patients had genetic testing, 52 patients (87%) were C282Y homozygotes. Median follow-up was 9.2 years (range 0 to 30 years). Nineteen patients (20%) developed hepatocellular carcinoma, one of whom was still living following transplantation. Cumulative survival for all patients was 88% at one year, 69% at five years and 56% at 20 years. Factors associated with death on multivariate analysis included advanced Child-Pugh score and hepatocellular carcinoma. Patients with hepatocellular carcinoma were older at the time of diagnosis of cirrhosis (mean age 61 and 54.6 years, respectively; P=0.03). The mean age at the time of diagnosis of hepatocellular carcinoma was 70 years (range 48 to 79 years). No other differences were found between the groups. CONCLUSIONS: Patients with hereditary hemochromatosis and cirrhosis are at significant risk of developing hepatocellular carcinoma. These patients are older when diagnosed with carcinoma and may have poorer survival following transplantation than patients with other causes of liver disease. Early diagnosis and treatment of hereditary hemochromatosis by preventing the development of cirrhosis may reduce the incidence of hepatocellular carcinoma in the future.     

肝癌组织中survivin、VEGF的表达及意义

目的探讨survivin与血管内皮生长因子（VEGF）在肝细胞肝癌（HCC）组织中的表达及其临床意义。方法采用免疫组化法检测50例HCC组织和20例正常肝组织中survivin、VEGF蛋白的表达,并分析其与HCC临床病理参数的关系。结果50例HCC组织中survivin、VEGF蛋白的阳性表达率分别为64.0%、68.0%,在正常肝组织中的表达率分别为0、10%,两者相比,P均〈0.05;survivin、VEGF蛋白表达与HCC临床分期及淋巴结转移相关（P〈0.05）;HCC中survivin、VEGF蛋白表达呈正相关（r=0.886,P〈0.05）。结论survivin和VEGF蛋白在HCC中表达率较高,为肝癌的分子靶向治疗提供了新的靶点;survivin和VEGF在HCC中的表达关系密切,对肝癌的发展可能有协同作用;测定HCC中survivin、VEGF蛋白的表达,对判断患者预后及指导治疗有较大帮助。     

Detection of telomerase activity in hepatocellular carcinoma by fluorescence-based TRAP method

Telomeres are important for the function and stability of eukaryotic chromosomes. Telomeric DNA is shortened every time liver cells divide. Normal human liver cells have a limited proliferative capacity, but immortalized cells prevent the shortening of the end of chromosomes by the expression of telomerase, the enzyme that elongates telomeric DNA. Although many genetic alterations have been reported in hepatocellular carcinoma, whether all hepatocellular carcinoma are immortalized cells is not known. Therefore, the telomerase activity was analyzed in 38 frozen samples from human hepatocellular carcinoma by fluorescence-based TRAP Method. Telomerase activity was detected in 34 (89.47%) of 38 hepatocellular carcinoma tissues regardless of tumor size, but telomerase activity was detected in only 12 (31.58%) of 38 nontumor liver tissues from patients with hepatocellular carcinoma. Diagnosis of hepatocellular carcinoma is sometimes difficult when the hepatomas are small and are of the differentiated type. Our results showed that the expression of telomerase may be of great value in diagnosing hepatocellular carcinoma.     

联合肝脏离断和门静脉结扎的二步肝切除术在不可切除肝细胞癌中的应用

对于残余肝体积不足的肝细胞癌患者,联合肝脏离断合并门静脉结扎的二步肝切除术(ALPPS)可以达到快速增大残余肝体积并切除肿瘤的目的,但仍有其弊端和未知性存在。ALPPS作为肝胆领域的新技术,在问世的10余年来,其适用范围在扩展,技术得到进一步改良。概述国内外对肝细胞癌患者施行ALPPS的经验,分析ALPPS在肝细胞癌患者应用中的关键问题,探讨其在靶向治疗和免疫治疗盛行时代中的机遇和挑战。