日本血吸虫感染致肝纤维化家兔肝脏微循环病变观察

目的 研究日本血吸虫感染致肝纤维化家兔的肝脏微循环病变。方法 对30只日本血吸虫感染致肝纤维化家兔和10只正常家兔取肝组织行光镜和电镜观察肝脏微循环病变。结果 HE染色示汇管区可见虫卵沉积，虫卵周围炎性细胞浸润。Mallory三色染色见肝内门静脉、肝动脉管壁纤维结缔组织增多，中膜平滑肌增殖肥大。透射电镜下见肝窦内皮受损严重，细胞大多崩解、破裂，窦内可见炎性细胞浸润，毛细胆管扩张，肝细胞肝窦面的微绒毛减少或断裂，肝窦呈毛细血管化。结论 肝脏微循环障碍是形成血吸虫肝纤维化门脉高压的发病基础之一。     

慢性乙型肝炎患者血清病毒标志和丙氨酸转氨酶与肝组织病理的相关性

目的 探讨慢性乙型肝炎（CHB）患者血清病毒标志和丙氨酸转氨酶（ALT）与肝组织病理的关系。方法 检测133例CHB患者血清病毒标记物和肝功能,同时全部患者接受彩色B超引导下快速经皮肝穿刺,将患者按HBeAg、HBV-DNA阳性与否各分为4个组．对各组的肝组织炎症和纤维化程度进行比较。结果 所有患者肝组织学显示肝内均有炎症、坏死及不同程度的纤维化存在。血清ALT正常的患者,作肝脏活组织病理检查．仍有一部分肝内有不同程度的炎症及纤维化改变,有的甚至存在肝硬化;ALT异常、HBeAg阴性的CHB患者肝脏炎症和纤维化程度更严重。血清病毒复制的活跃程度与肝组织病理损害的程度不成正比。结论 单凭血清ALT升高、血清病毒复制活跃与否判断肝病活动性是不全面的,应将肝组织病理作为判断肝炎活动性和是否抗病毒治疗的主要依据。     

肝窦内皮细胞及其与肝细胞肝癌关系的研究进展

肝窦内皮细胞是一组在形态结构和功能方面有别于血管内皮细胞的具有高度异质性的群体,在正常肝组织中表达CD4、CD14、CD32、NO合酶、内皮素-1等因子。在肝细胞肝癌的发生、发展过程中肝窦发生毛细血管化,在此过程中肝窦内皮细胞显著表达Ⅷ因子相关抗原、CD34等重要标记物。     

肝细胞、肝窦内皮细胞分离培养及肝细胞支架的制备

目的：探究大鼠肝细胞、肝窦内皮细胞的分离培养方法及肝去细胞支架的制备方法。方法：用胶原酶IV消化肝脏组织后经洗涤、过滤、离心等步骤分离出大鼠肝细胞,然后置于培养基内进行纯化、培养后行细胞免疫组织化学检测;通过Percoll密度梯度离心法分离出肝窦内皮细胞置于培养基内纯化、培养后行免疫荧光检测;肝组织经循环灌注去细胞处理制备成大鼠肝细胞去细胞支架后经扫描电镜证实。结果：分离出的大鼠肝细胞在显微镜下呈光亮圆形,饱满,胞核清晰。肝细胞细胞角蛋白18免疫组织化学染色后见胞浆内存在棕黄色颗粒。肝窦内皮细胞呈梭形,细胞核位于中央。内皮素-1免疫荧光染色后细胞表面呈红色信号。制备成的大鼠肝脏去细胞支架在扫描电镜下显示细胞外基质保留,无细胞核及细胞质成分。结论：成功分离出了肝细胞和肝窦内皮细胞并成功制备了肝去细胞支架。     

HBeAg阴性和阳性慢性乙型肝炎病人肝脏组织学变化

目的比较HBeAg阴性和阳性慢性乙型肝炎病人肝脏组织学变化情况。方法对266例行肝活检的慢性乙型肝炎病人行免疫组化检测肝组织HBsAg、HBcAg的变化;采用双抗体夹心酶联免疫法检测血清前S1蛋白、HBV—DNA、HBsAg等的变化。按肝组织炎症、纤维化程度分组,比较HBeAg阳性和阴性的分布,并比较肝组织中HBsAg、HBcAg在HBeAg阳性和阴性组的阳性率。结果HBeAg阳性和阴性组肝组织炎症程度分布比较差异有显著性（Uc=2．716,P〈0．01）。HBeAg阳性和阴性组肝组织纤维化程度分布比较差异无显著性（Uc=1．493,P〉0．05）。HBeAg阳性组血清前S。蛋白、HBV—DNA及肝组织中的HBsAg、HBcAg的阳性率明显高于HBeAg阴性组,差异有极显著性（χ^2=8．47～41．29,P〈0．01）。结论HBeAg阳性组肝组织炎症程度高于阴性组。HBeAg阳性组血清前s。蛋白、HBV—DNA及肝组织中的HBsAg、HBcAg等病毒复制指标的阳性率明显高于HBeAg阴性组。     

慢性乙型肝炎患者外周血单个核细胞及肝组织中HBVcccDNA的相关性

目的：通过对慢性乙型肝炎患者肝组织及外周血单个核细胞（PBMC）中乙肝病毒共价闭合环状DNA（HBVcccDNA）的特异性定量检测,探讨两者的相关性和临床可行性。方法：取本院慢性乙型肝炎患者肝组织穿刺标本20份,在肝穿刺当天抽取患者外周抗凝血5mL,标本经相应处理后行HBVcccDNA检测。结果：20份慢性乙型肝炎患者肝组织标本中有18份HBVcccDNA检测为阳性,最大值为5．61×10^5copy／μg,最小值为3．26×10^3copy／μg,平均为1．79×10^4copy／μg。20份血浆中PBMC中HBVcccDNA检测均为阴性。结论：在慢性乙型肝炎患者肝纽织中可检测到HBVcccDNA,但未能在PBMC中检测到HBVcccDNA,因此PBMC不能支持HBV的复制。     

内皮素-1诱导大鼠门脉高压以及丹酚酸B干预过程中肝窦内皮的变化

目的:探讨肝窦内皮在内皮素-1(ET-1)诱导大鼠门脉高压以及丹酚酸B干预过程中的变化。方法:将20只SD大鼠随机分为4组,均采用门静脉穿刺灌流。在灌流缓冲液5min后,预防对照组予灌流ET-1缓冲液,中药预防组予丹酚酸B ET-1灌流液,治疗对照组予ET-1液灌流及缓冲液,中药治疗组予ET-1及丹酚酸B灌流液。分别观察4组大鼠各时段门脉灌流压后处死取材,通过形态学方法,观察ET-1诱导大鼠门脉压升高及丹酚酸B干预的过程中肝窦内皮窗孔的变化。结果:在ET-1诱导大鼠门脉灌流压升高过程中,肝窦内皮细胞窗孔体积缩小,数量减少;用含丹酚酸B的缓冲液灌流后,大鼠门静脉灌流压较对照组低,肝窦内皮细胞窗孔体积变大,数量增多。结论:肝窦内皮在ET-1诱导大鼠门脉压升高的过程中可能起着重要的作用;活血化瘀药丹参的有效成分丹酚酸B能有效预防或改善由ET-1诱导的肝窦内皮细胞失窗孔及门脉压的升高。     

Hepatitis with Australia antigenemia following renal transplantation

Over a seven-year period 18 of 125 patients who underwent renal transplantation developed hepatitis. Acute hepatic necrosis occurred in two, chronic aggressive hepatitis progressing to posthepatitic cirrhosis in eight, chronic persistent hepatitis in five, acute hepatitis with recovery in two and cholestatic hepatitis in one. Hepatic failure was the cause of death in four and a major contributing factor in three. Fifteen of the 18 were of blood Group A. After renal transplantation Australia antigen (Au) was present in the blood of 12 of the 15 patients with hepatitis who were tested and in one of 38 patients without clinical evidence of liver disease. Once present, Au persisted in all patients but one. Particles measuring 210 to 250 Å, characteristic of Au, were seen in liver cells by electronmicroscopy in nine of the 10 patients examined who had hepatitis with Australia antigenemia, but they were not seen in the two patients studied with Au-negative hepatitis.     

原发性肝癌血管起源的研究

目的 研究原发性肝癌的血管起源。方法 用病理及CD34、荆豆凝聚素（UEA-1）免疫组化的方法对21例肝癌及癌旁组织中血管形成进行检测和观察。结果 癌旁组织血窦内皮细胞CD34及UEA-1染色阴性, 炎症组织内的血管染色阳性。肿瘤血管与癌旁组织血管相连或不相连。结论 部分原发性肝癌血管可能起源于毛细血管化的癌旁肝血窦。     

丙型肝炎病毒C33抗原在肝组织中的表达

目的：观察ＨＣＶＣ３３抗原在慢性丙型肝炎患者肝组织中的分布。方法：４例慢性丙型肝炎患者肝组织用免疫组化法作ＨＶＣＣ３３标记。结果：ＨＣＶ３３抗原不仅分布于肝细胞核，胞浆及肝细胞膜中，而且分布肝窦内皮细胞及肝内脉巴细胞的胞浆及胞膜上。结论：ＨＣＶ抗原存在于上述肝细胞内，是慢性丙型肝炎的重要标志。     

慢性乙型肝炎患者肝组织及外周血单核细胞内IFN-α/β受体的表达及其临床意义

目的 慢性乙型肝炎患者肝组织的炎症程度是影响干扰素应答率的独立因素,本研究的目的 系从干扰素受体的角度来探讨其机制,此外了解外周血单核细胞干扰素受体与慢性乙型肝炎感染的关系.方法 采用链霉抗生物素蛋白-过氧化物酶连接法,检测15健康对照、16例慢性乙型肝炎患者外周血单核细胞及另外21例慢性乙型肝炎患者肝组织内的干扰素受体的表达并进行定量分析.结果 15例健康对照、16例慢性乙型肝炎患者外周血单核细胞干扰素受体的表达量分别为(21.4060±4.7658)、(23.8365±3.3329)(P=0.374);另外21例慢性乙型肝炎患者根据肝脏病理的炎症程度分为3组G1(5.6913±1.8422)、G2(7.4706±5.3572)、G3(25.1307±7.0700)(P=0.000).结论 慢性乙型肝炎患者肝组织内干扰素受体的表达量与肝组织的病理炎症程度相关,可能是其对干扰素应答率高的原因,而外周血单核细胞干扰素受体的表达与HBV感染无关.     

慢性乙型肝炎患者HBV DNA载量与肝组织病理的关系

目的探讨慢性乙型肝炎（CHB）患者血清中HBV DNA含量与肝组织中HBV DNA含量的相关性以及血清HBV DNA水平与肝组织病理的关系。方法选取70名未经免疫或抗病毒治疗的CHB患者。用荧光PCR法检测CHB患者血清、肝组织中HBV DNA的含量，并与同期肝组织活检病理结果对比分析。结果肝组织中HBV DNA载量明显高于血清HBV DNA载量，但两者呈高度相关（r=0．882，P〈0．05）；肝组织病理分级与血清HBV DNA含量之间正相关关系（r=0．210，P〈0.05）。结论肝组织HBV DNA水平较血清HBV DNA水平更能准确反映HBV DAN复制情况；随着血清HBV DNA浓度的增高，肝细胞炎症及肝细胞纤维化程度也随着加重。     

人卵巢癌微血管内皮细胞的培养与鉴定

目的建立人卵巢癌微血管内皮细胞（ovarian carcinoma-derived microvascular endothelial cells,ODMECs）体外培养体系。方法采用胶原酶、胰酶联合消化法分离微血管内皮,经pereoll梯度密度离心纯化。光镜、电镜、流式细胞术、免疫细胞化学对所获得的ODMECs进行鉴定。结果所获0DMECs流式分析有内皮标志物CD34／VEGF—R2表达;免疫荧光FⅧ一RAg阳性;电镜下见细胞多核、有丰富的微丝、weibel—Palade小体等;培养中的内皮细胞生长状态良好、呈现典型的铺路石征,可传代培养。结论本研究建立了人0DMECs体外培养体系,对了解卵巢癌血管内皮的异质性有重要价值,为抗卵巢癌血管生成的研究奠定了基础。     

基质金属蛋白酶-2在肝硬化患者肝组织中的表达

目的了解基质金属蛋白酶-2(MMP-2)在肝组织中的表达情况及与肝硬化患者肝功能损害程度的关系。方法采用免疫组织化学方法(SP法)观察了7例正常肝组织和23例肝炎后肝硬化患者肝组织中MMP-2表达的情况。结果在肝硬化患者肝组织中发现MMP-2主要在肝细胞、血管内皮细胞、肝窦内皮细胞及胆管内皮细胞的胞浆中高表达,表达情况与肝硬化Child-Pugh A、B、C分级相关,随着Child-Pugh分级及纤维化程度增高,MMP-2表达逐渐减少,A、B级患者肝组织中MMP-2表达最多,与对照组比较明显增高(P<0.01),A、B级与C级之间比较均有统计学意义(P<0.05)。结论肝硬化Child-Pugh分级不同,MMP-2表达发生了不同的变化,提示MMP-2在肝硬化形成中有重要作用。     

Clinical pathological study of treatment of chronic hepatitis with hyperbaric oxygenation

Objective To detect the feasibility and theoretic basis for treatment with hyperbaric oxygenation (HBO) in chronic hepatitis and to compare the changes in hepatic function, immunity, pathologic morphology, ultrastructure and HBV in hepatic tissues before and after treatment. Methods Sixty cases of chronic hepatitis were randomly selected and divided into two groups: the experiment (n=30) and control groups (n=30). Patients in the experimental group were treated with HBO for 6 courses. Patients in the control group were treated for 60 days with the usual drugs used in the clinic. The function and bloodstream graph of liver were examined and liver biopsies were made before and after treatments. Routine paraffin sections were stained with HE and observed under the light microscope. Ultra thin slides from paraformaldehyde and glutaraldehyde fixed liver tissue were stained with lead citrate and observed with the transmission electric microscope. HBsAg and HBcAg in liver of the experimental group were detected with ABC immunohistochemistry method before and after treatment. Results For the experimental group, ALT, SB, γ-GT, AKP, IgG, and IgM in blood and the degeneration and necrosis of hepatocytes were remarkably decreased (P&lt;0.05), the mean contractive wave of bloodstream in liver and the bloodstream in right ramus of janitrix were remarkably increased (P&lt;0.05), and the swelling of mitochondria, increase of lysosomes, generation of Kupffer cells, infiltration of lymphocytes in portal area and capillary generation were all remarkably alleviated (P&lt;0.05) after treatment with HBO. There were significant differences between the experimental and control groups after treatment with different methods (P&lt;0.05). For patients in the experimental group, the fibrosis and fat-storing cells in the liver were not reduced (P&gt;0.05), and the expression of HBsAg and HBcAg in liver was not weakened (P&lt;0.05) after treatment. Conclusions Treatment with HBO for chronic hepatitis was effective and recommendable, but it could not reverse liver fibrosis. However, it might be able to delay or prevent the liver from fibrosis, so it might be more effective at the early and middle stages of chronic hepatitis. HBO could not inhibit the HB virus. So we consider that treatment with HBO should be simultaneous with anti HBV therapy.     

Observation of T Lymphocyte Subsets in the Liver of Patients with Advanced Schistosomiasis and Advanced Schistosomiasis Accompanied with Hepatitis B

It has been demonstrated that severe liver damage and aggravated clinical manifestations and poorprognosis might take place in AS patients accompanied with hepatitis B[l--SJ. The status of cellular immune response in the liver, however, has not beenavailable in the literatures about the disease up tonow. In this study, T lymphocyte subsets in the liverwere detected by immunohistochemical technique inAS patients and those with AS accompanied withhepatitis B. The distribution of T lymphocyt…     

血清GP73作为原发性肝癌肿瘤标志物的来源探究

目的探究血清GP73作为原发性肝癌的肿瘤标志物的可行性。方法采用SYBR Green qPCR法对103例原发性肝癌（PHC）、13例肝硬化、49例肝炎和50例健康人群外周血单核细胞GP73 mRNA的相对表达量进行检测,同时检测分析8份正常肝组织和8份肝癌组织的GP73 mRNA相对表达水平。对7例病理确诊为PHC的肝组织和1例正常肝组织、1例肝硬化肝组织中GP73的表达进行免疫组化分析。采用ELISA法检测血清GP73水平和利用电化学发光法血清中AFP水平。结果外周血有核细胞GP73 mRNA含量四组间差异无统计学意义（F=3.013,P=0.055）。肝癌组织GP73mRNA表达量（12.64）显著高于正常肝组织（1.00）。GP73免疫组化结果显示,正常肝组织中GP73少量在肝实质细胞胆小管处表达。肝硬化组织中GP73在肝细胞胞浆中弱阳性表达,但在肝细胞胞浆的两侧靠近胆小管位置呈颗粒状浓集;胆管柱状上皮也呈强阳性着色。PHC患者肝组织GP73在肿瘤细胞胞浆呈强阳性着色,且在胆管上皮细胞靠近腔缘一侧呈深棕色颗粒状分布。肝癌组中血清GP73水平显著高于肝炎组及健康对照组（χ^2=62.541,P=0.000）。通过ROC曲线确定诊断肝癌的GP73临界值为113.2μg/L,和AFP的临界值为13.3μg/L时,GP73和AFP单项检测的敏感度分别为68.0%和56.3%,特异度分别为94.8%和91.8%。结论 PHC患者血清高表达的GP73可能是在癌变的肝细胞胞浆中合成随后分泌入血。血清GP73在诊断PHC时阳性检出率显著高于AFP,且ROC曲线提示灵敏度和特异度均优于AFP。     

Detection of hepatitis B virus DNA in mononuclear blood cells

The Southern transfer hybridisation technique was used to test mononuclear blood cells for hepatitis B virus DNA. Viral DNA sequences were detected in mononuclear cells of 10 out of 16 patients with hepatitis B virus infection and in none of 21 normal controls. Blood contamination was excluded by the absence of hepatitis B virus DNA in the corresponding serum samples in all cases. Free monomeric hepatitis B virus DNA was found in three patients positive for hepatitis Be antigen (HBeAg) and one positive for anti-HBe, and integrated hepatitis B virus DNA was present in four patients positive for anti-HBe. In two other patients the small size of the samples did not allow a distinction between free and integrated viral DNA. The state of the virus in the mononuclear cells seemed to correlate with the HBeAg or anti-HBe state, as has been noted in the liver. These results indicate that hepatitis B virus may infect mononuclear blood cells, thereby expanding the tissue specificity of this agent beyond the liver, as has been reported for pancreatic, kidney, and skin tissue. They also suggest that hepatitis B virus infection of mononuclear cells might be related to immunological abnormalities observed in carriers of the virus.     

Fas在慢性丙型肝炎肝组织中的表达

目的通过检测慢性丙型肝炎患者肝组织Fas的表达,探讨Fas-FasL系统在慢性丙型肝炎发病机制中的作用及其临床意义。方法50例慢性丙型肝炎患者肝脏穿刺取肝组织做病理活检。免疫组织化学法检测肝组织中Fas的表达,分析肝组织中Fas表达与肝组织炎症、纤维化及临床分型间的关系。结果慢性丙型肝炎患者肝组织细胞中Fas表达随着肝脏炎症活动度和肝纤维化程度加重而增加(P<0.05),随着临床病情的加重而增加(P<0.05)。结论Fas-FasL系统与慢性丙型肝炎患者肝组织的炎症和纤维化有关,Fas-FasL系统介导的肝细胞凋亡可能是慢性丙型肝炎发病机制之一。