斑点杂交法检测TTV DNA

用ＴＴＶ基因组ＯＲＦ１区段的引物进行ＰＣＲ扩增，扩增产物用地高辛标记的特异性探针进行斑点杂交检测。结果检测了５４３例肝炎病人和献血员血清标本中ＴＴＶＤＮＡ，阳性检出率为１８．４％（１００/５４３），其中甲型肝炎阳性率为６．０％（２/３３），乙型肝炎为７．８％（５/６４），丙型肝炎为２．３％（１/４３），非甲－戊型肝炎为３９６％（４０/１０１），输血后肝炎为１２．５％（５/４０），献血员为１７．９％（４７     

血液透析患者输血传播病毒感染状况的初步研究

目的：研究血液透析（ＨＤ）患者输血传播病毒（ＴＴＶ）的感染状况和基因型。方法：设计特异性引物,建立工聚合酶链反应,对２０例ＨＤ患者血清标本进行ＴＴＶ－ＤＮＡ的检测。并对阳性ＰＣＲ产物进行荧光法直接测序。结果：经过巢式ＰＣＲ检测,ＨＤ患者ＴＴＶ－ＤＮＡ阳性率为２５％。随机选择ＴＴＶ－ＤＮＡ阳性ＰＣＲ产物进行荧光法直接测序,与国外报道的日本株Ｎ２２比较,核苷酸同源性为９５．８％。     

原发性肝癌组织HBVDNA存在状态及HBsAg,HBcAg表达的检测

本文采用Ｓｏｕｔｈｅｒｎ印迹杂交技术检测原发性肝癌（ＰＨＣ）组织ＨＢＶＤＮＡ存在状态，同时应用免疫组化染色检测ＰＨＣ组织内ＨＢｓＡｇ、ＨＢｃＡｇ表达，并比较ＨＢｓＡｇ、ＨＢｃＡｇ在ＰＨＣ组织与癌旁肝组织的定位分布。结果显示：整合型ＨＢＶＤＮＡ在３２例ＰＨＣ组织检出率７１．９％（２３／３２），癌旁组织检出率４０．７％（１１／２７）；ＰＨＣ组织内检出ＨＢＳＡｇ阳性细胞１２例（３７．５％）未检出ＨＢＣＡｇ；癌组织检出ＨＢｓＡｇ２６例（９６．３％），ＨＢｃＡ９５例（１８．５％）。结果提示：ＨＢＶ感染与ＰＨＣ发生密切相关，ＨＢＶ可能通过整合于宿主细胞染色体对ＰＨＣ发生起一定作用。     

血液透析病人中输血传播病毒（TTV）感染的检测及序列分析

目的：检测我国南方地区血液透析的病人中经输血传播病毒（ＴＴＶ）的感染。方法：采用ＴＴＶ基因组ＯＲＦ１区段设计特异性引物,建立巢式ＰＣＲ方法检测４８例血液适析病人血清标本中ＴＴＶ ＤＮＡ,并对分离出的２例ＴＴＶ部分基因进行核苷酸序列分析。结果：血液透析病人ＴＴＶＤＮＡ阳性率为４１．７％（２０／４８）,２例ＴＴＶＤＮＡ基因序列与日本ＴＴＶ基因相对应位置的核苷酸同源性分别为９２％和９５％。结论：我国南方     

用微板核酸杂交-ELISA技术检测皖北地区不同人群TTV感染情况

目的：用微板核酸杂交－ＥＬＩＳＡ技术检测皖北地区不同人群ＴＴＶ感染情况。方法：用ＰＣＲ法将患者血清标本中的ＴＴＶ ＤＮＡ扩增后,与两条特异性探针夹心杂交,通过酶联显色检测血甭标本中的ＴＴＶ ＤＮＡ,进行分型并与套式ＰＣＲ方法比较。结果：ＴＴＶ在学生、血透患者、肝病患者中的阳性率分别为３．３％,１６．７％和１３．５％,前者与后两者比较差异均有显著意义（Ｐ〈０．０５）;１４例非甲－非戊（ＮＡ－ＮＥ）肝     

聚合酶反应检出石蜡包埋肝癌组织的乙型肝炎病毒DNA

传统免疫组织化学方法及分子杂交检测石蜡包埋肝组织乙型肝炎病毒（ＨＢＶ）ＤＮＡ灵敏度低，聚合酶链反应（ＰＣＲ）方法能否检验到石蜡肝癌组织中的ＨＢＶ ＤＮＡ报道尚不见。为此，采用ＨＢＶ与丙型肝炎病毒二合－ＰＣＲ试剂盒对１７例肝癌患者的石蜡包埋癌组织进行了ＨＢＶ ＤＮＡ检测，结果７例阳性，阳性率为４１％，而８例非肝癌对照患者则均阴性。组织ＨＢＶ ＤＮＡ结果与血清ＨＢｓＡｇ比较显示，该试剂盒有较高的检出率     

83例静脉药瘾者的TT病毒感染状况

目的：调查静脉药瘾者中ＴＴ病毒（ＴＴＶ）的感染状况。方法：在ＴＴ病毒开放读码框（ＯＲＦ）１保守区设计引物,建立巢式ＰＣＲ,检测８３例静脉药瘾者和１０５名献血员血清中ＴＴＶ ＤＮＡ。结果：８３例静脉药瘾者中３３例ＴＴＶ ＤＮＡ阳性,阳性率为４０％,而献血员的阳性率为１１％（Ｐ〈０．０５）。结论：静脉药瘾者是ＴＴ病毒感染的高危人群,不洁注射是感染ＴＴ病毒的重要途径。     

聚合酶链反应检出石蜡包埋肝癌组织的乙型肝炎病毒DNA

传统免疫组织化学方法及分子杂交检测石蜡包埋肝组织乙型肝炎病毒（ＨＢＶ）ＤＮＡ灵敏度低，聚合酶链反应（ＰＣＲ）方法能否检测到石蜡肝癌组织中的ＨＢＶＤＮＡ报道尚不多见。为此，采用ＨＢＶ与丙型肝炎病毒二合一ＰＣＲ试剂盒对１７例肝癌患者的石蜡包埋癌组织进行了ＨＢＶＤＮＡ检测，结果７例阳性，阳性率为４１％，而８例非肝癌对照患者则均阴性。组织ＨＢＶＤＮＡ结果与血清ＨＢｓＡｇ比较显示，该试剂盒有较高的检出率及符合率。这为今后进一步研究大量保存的肝病石蜡包埋组织的ＨＢＶ感染提供了一个可靠灵敏的方法，也为研究其他病毒感染的石蜡包埋材料提供了参考。     

249例尖锐湿疣及湿疣样病变形态学及HPV原位杂交对照观察

收集２４９例临床诊断或怀疑为尖锐湿疣的活检标本作病理形态学观察，组织学诊断为尖锐湿疣１４８例，可疑１８例，炎性增生８３例。抽取其中３０例尖锐湿疣、１０例可疑尖锐湿疣、１５例炎性增生作地高辛标记的ＨＰＶ６Ｂ／１１ＤＮＡ原位杂交。结果：尖锐湿疣组均阳性，可疑组１例阳性，炎性增生组均阴性。本文对尖锐湿疣的病理诊断及鉴别诊断进行了讨论。本组材料中的９２．８％的病例在组织学上得到了确诊，并且和ＨＰＶ６Ｂ／１１原位杂交结果相符，仅７．２％病例组织学列为可疑，需进一步作ＨＰＶ检测。     

高危人乳头瘤病毒,p16,p53蛋白在胃癌和癌前病变中的 …

目的 探讨高希ＨＰＶ感染与抑癌基因表达产物ｐ５３、ｐ１６蛋白在胃癌发生、发展中的作用及与肿瘤预后的关系。方法 应用原位杂交和免疫组化技术，分析９６例胃癌和癌前病变及腺癌组织中ＨＰＶ－１６／１８ＤＮＡ、ｐ５３和ｐ１６蛋白的阳性表达情况。结果ＨＰＶ－１６亚型和１８亚型的阳性率分别为３４．３８％和８．３３％；ｐ５３和ｐ１６蛋白阳性表达率分别为４６．８８％和４３．７５％。ＨＰＶ－１６亚型、ｐ５３和ｐ１６     

Hepatic allograft rejection. Diagnostic and therapeutic aspects]

Rejection is still a major problem in liver transplantation: 50-70% of patients present at least one acute episode, while 5-15% develop chronic rejection. Acute rejection is suggested by clinical signs and abnormal laboratory test results, but only histological signs on biopsy specimens are adequately specific. The three most frequent elementary lesions are a portal infiltrate, bile duct alterations and endothelial inflammation. Several systems have been forwarded to classify the degree of rejection. Chronic rejection is characterized by a progressive reduction in the number of interlobular bile ducts. It does not respond to available immunosuppressive drugs and thus requires retransplantation. Prophylactic immunosuppression is usually based on the triple-drug combination cyclosporine/azathioprine/corticosteroids. The orthoclone OKT3 and FK506 have recently been proposed for prophylactic use in this setting. Treatment of liver graft recipients with cyclosporine carries a number of specific disadvantages, particularly with regard to gastrointestinal interactions (delayed intestinal absorption in patients with cholestasis, bile derivation, diarrhea or receiving cholestyramine; possibility of a sharp increase in blood cyclosporine levels when the Kehr drain is clamped), and drug interactions (cyclosporine metabolization of CP450-IIIA accounts for most such interactions). The results of radioimmunoassay and TDx must be interpreted with care, and HPLC remains the reference technique in borderline cases. Cyclosporine is hepatotoxic in about 20% of cases, generally giving rise to cholestasis. This toxicity is dose-dependent and therefore diminishes when the dosage is reduced. Most groups initially treat rejection with corticosteroids. The response to treatment is generally evaluated in terms of liver function tests.(ABSTRACT TRUNCATED AT 250 WORDS)     

输血传播病毒感染与丙型肝炎病毒感染的相互影响

目的了解输血传播病毒(TTV)与丙型肝炎病毒(HCV)重叠感染的发生率,探讨TTV感染与HCV感染的相互影响。方法采用酶联免疫吸附试验(ELISA)法检测血清中人类免疫缺陷病毒(HIV)、乙型肝炎病毒(HBV)、HCV、庚型肝炎病毒(HGV)及TTV标志物,应用巢式聚合酶链反应(n-PCR)技术检测血清TTV DNA,用速率法或终点法检测血清肝功能指标,用放射免疫法检测血清肝纤维化指标,用超声诊断仪检查肝胆脾形态及动态指标;应用SPSS 11.0软件分析比较肝功能检测结果、肝纤维化指标检测结果及肝脾形态和动态指标改变的差异。结果TTV/HCV重叠感染占TTV感染的69.6%(39/56),占HCV感染的61.9%(39/63)。TTV、HCV感染与TTV/HCV重叠感染肝功能检测结果差异有统计学意义(P<0.05);肝纤维化指标检测结果差异有统计学意义(P<0.05)。结论TTV/HCV重叠感染存在很高的发生率,感染者肝损程度较重,临床进程加快,有肝纤维化趋势。     

Histological findings of TTV-positive non-B non-C chronic hepatitis patients]

TT virus has been reported in association with patients with acute and chronic liver disease of unknown etiology. In order to estimate the pathogenesis of TTV, we investigated the liver histology in the patients of TTV-positive chronic hepatitis. The findings frequently observed in TTV-DNA positive cases were changes in liver parenchyma such as focal necrosis, hepatocyte degeneration or fatty change of various degree. On the other hand, degree of chronic inflammation in portal areas such as lymphocyte infiltration, piecemeal necrosis or fibrosis were relatively mild, which might suggest the good prognosis in TTV-positive cases. However, we could not find any differences in liver histology between TTV positive and negative patients.     

Hepatic adenine nucleotides and microsomal cholesterol 7α-hydroxylase activity in the obstructed and freely draining lobes of the liver after selective bile duct obstruction

Background: The effect of selective bile duct obstruction (SBDO) on hepatic reserve function of the bile duct obstructed (BDO) and nonobstructed freely draining (FD) lobes of the liver is obscure.Methods: The bile duct branches draining from the left lateral and median lobes of the liver were ligated for 4 and 10 days in rats, and hepatic reserve functions in BDO and FD lobes were assessed by microsomal cholesterol 7α-hydroxylase activities and by hepatic adenine nucleotides and energy charge levels. The values were compared with those in the sham-operated control liver. Cholesterol 7α-hydroxylase activities were determined by gas-liquid chromatography-mass spectrometry, and hepatic adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) levels with high-pressure liquid chromatography.Results: The histological examination of the BDO lobes showed proliferation and formation of new bile ductules and fibrous connective tissues linking portal areas. Microsomal cholesterol 7α-hydroxylase activities, hepatic energy charge and each adenine nucleotide level did not differ between FD and BDO lobes, and the values were similar to those in the sham-operated liver.Conclusions: Selective bile duct obstruction shows no adverse effects on microsomal and mitochondrial functions in both the BDO and FD lobes of the liver.     

Laparoscopic histopathological analysis of "gentle undulation" findings observed in patients with primary biliary cirrhosis

Laparoscopic findings of the liver in 13 cases with primary biliary cirrhosis (PBC) diagnosed by wedge or needle biopsy were investigated. The characteristic features of laparoscopic appearance--gentle undulation--were observed in 11 out of 13 (85%) patients with PBC. These gentle undulations were irregularly shaped areas from 0.5 to 3 cm in diameter. Those observed in s-PBC were greater in number and more pronounced than those in a-PBC. In the concave areas of these undulations, the number of portal tracts was significantly greater than in the convex areas (p less than 0.005). On the contrary, the number of interlobular bile ducts in the concave area was significantly less than in the convex area (p less than 0.005). Therefore, gentle undulation was relatively specific to PBC, and was caused by scar formation and chronic non-suppurative destructive cholangitis in the portal area.     

Reconstruction of intrahepatic bile ducts in congenital biliary atresia.

Macroserial reconstruction of the main intralobular bile ducts was made in 7 cases of biliary atresia; 2 cases of type I, 1 case of type II and 4 cases of type III according to Kasai's classification. From the results of these reconstruction studies, it was confirmed that the main interlobular bile ducts are usually patent through the liver regardless of the type of atresia of the extrahepatic bile ducts. A microserial reconstruction of bile ducts and ductules of a small portal tract performed in one case disclosed that a number of ductules make a network around the main duct and have some communications with the main duct. These results were compatible with the fact that active excretion of bile was obtained in many postoperative patients with biliary atresia. As observed in one case of the present series, postoperative complication of severe ascending cholangitis seemed to be an important cause of destruction or disappearance of intrahepatic bile ducts, which has also seen in older infants with this disease without complicaion of cholangitis. In view of these facts the operation of an early stage of life is recommended in biliary atresia.     

Primary sclerosing cholangitis and hepatitis C virus infection

Two cases of primary sclerosing cholangitis with hepatic C virus infection in a 62-year-old man and a 60-year-old woman are presented. The infection in the man was eradicated with interferon therapy in 1992. Seven years thereafter, endoscopic retrograde cholangiography revealed a diffuse 2.5-cm-long stenotic lesion in the common bile duct which was consequently resected. Histological examination of the resected specimen revealed proliferation of epithelial cells, plasma cell infiltration, and fibrosis in the submucosal layer of the common bile duct. The human leukocyte antigen DR loci were 2 and 9. In the woman, a 6-month course of interferon therapy in 1992 failed to eradicate the infection. Cholangiography in 1999 revealed multiple narrowings and dilatations of intra- and extrahepatic bile ducts. Ultrasound guided biopsy of the liver in 1992 had revealed onionskin lesions around the bile duct epithelium in the portal tract. The human leukocyte antigen DR locus was 2. From these findings, the 2 cases were diagnosed as primary sclerosing cholangitis. Further studies may provide insights into the relation between the pathogenesis of the disease and the infection.     

术中B型超声检查在肝癌手术中的应用

85例肝癌病人在术中进行了术中B超检查,其结果与术前B超、CT进行了对照。作者发现(术中B超检查)对大的肝癌结节(主瘤)显示三者无统计学差异;对肝内转移灶(子瘤)显示分别为90.32％、22.58％、16.18％;门脉干支内癌栓显示为88.88％、22.22％、11.11％,对胆管内癌栓分别为83.33％、33.33％、33.33％。本组肿瘤切除68例,切除率80％,死亡率1.17％。术中B超检查可清晰显示癌周血管,有利于判断肿瘤能否切除,减少术中血管损伤,检出19例病人肝实质内子瘤31个,15例得到合理切除,小肝癌定位5例,顺利切除。     

The prevalence of TTV infection in non-A to G chronic liver disease, especially non-A to G hepatocellular carcinoma, and the clinical significance of TTV infection]

We examined the prevalence of TT virus (TTV) infection in non-B, non-C, non-G chronic liver disease, in particular, in hepatocellular carcinoma (HCC), and the clinical significance of TTV infection. Among 829 cases with chronic liver disease, 30 cases (4%) had non-B, non-C, non-G chronic liver disease. The percentage of TTV-DNA positive cases among these 30 cases with non-B, non-C, non-G chronic liver disease was 57% (17/30), significantly higher than the percentage TTV-DNA positivity (14%; 5/107) among blood donors (P < 0.01). All the TTV-DNA positive cases were still positive for TTV-DNA throughout the follow-up period (4 +/- 2 years; 1-7 years), thus demonstrating that TTV infection is persistent. These findings suggest that TTV may be one of the causes of non-B, non-C, non-G chronic liver disease. When the 30 cases of non-B, non-C, non-G chronic liver disease were divided into a TTV-DNA positive group and TTV-DNA negative group and the clinical data between the two groups compared, it was found that the serum ALP and serum gamma-GTP levels in some cases in the TTV-DNA positive group were higher than those in the TTV-DNA negative group. Twelve cases of non-B, non-C, non-G HCC were divided into two groups (TTV-DNA positive and TTV-DNA negative), and the clinical data between the two groups compared. All the four cases of HCC which were not associated with liver cirrhosis were TTV-DNA positive. However, with respect to the age at the time of onset of the HCC, no significant differences were noted between the cases of HCC associated with liver cirrhosis and those not associated with liver cirrhosis. In spite of older patients, many cases of HCC associated with TTV infection are not associated with liver cirrhosis.     

肝内胆管细胞癌的MSCT诊断和鉴别

目的提高对肝内胆管细胞癌(ICC)CT表现的认识和术前诊断准确率。方法收集2002-2010年14例经病理证实的ICC术前CT进行回顾性分析。结果 ICC平扫CT表现为低密度肿块或胆管扩张影,9例位于周围,4例为多发病灶,2例病灶周围见环形低密度影,11例病灶远端胆管扩张,2例临近肝缘萎陷,2例合并门静脉癌栓或淋巴结转移。增强扫描动脉期9例病灶不同程度强化,边缘强化为主,静脉期增加至11例,延迟期所有病灶均强化,呈渐进性向心性强化。本科室术前正确诊断ICC11例,诊断可靠率约78%。结论 MSCT三期增强检查可作为诊断ICC的重要手段。