Safety of hepatitis B virus core antibody-positive grafts in liver transplantation: A single-center experience in China

BACKGROUND Given the shortage of suitable liver grafts for liver transplantation, proper use of hepatitis B core antibody-positive livers might be a possible way to enlarge the donor pool and to save patients with end-stage liver diseases. However, the safety of hepatitis B virus core antibody positive(HBcAb+) donors has been controversial. Initial studies were mainly conducted overseas with relatively small numbers of HBcAb+ liver recipients, and there are few relevant reports in the population of China's Mainland. We hypothesized that the safety of HBcAb+ liver grafts is not suboptimal.AIM To evaluate the safety of using hepatitis B virus(HBV) core antibody-positive donors for liver transplantation in Chinese patients.METHODS We conducted a retrospective study enrolling 1071 patients who underwent liver transplantation consecutively from 2005 to 2016 at West China Hospital Liver Transplantation Center. Given the imbalance in several baseline variables, propensity score matching was used, and the outcomes of all recipients were reviewed in this study.RESULTS In the whole population, 230 patients received HBcAb+ and 841 patients received HBcAb negative(HBcAb-) liver grafts. The 1-, 3-and 5-year survival rates in patients and grafts between the two groups were similar(patient survival: 85.8% vs 87.2%, 77.4% vs 81.1%, 72.4% vs 76.7%, log-rank test, P = 0.16; graft survival: 83.2% vs 83.6%, 73.8% vs 75.9%, 70.8% vs 74.4%, log-rank test, P = 0.19). After propensity score matching, 210 pairs of patients were generated. The corresponding 1-, 3-and 5-year patient and graft survival rates showed no significant differences. Further studies illustrated that the post-transplant major complication rates and liver function recovery after surgery were also similar. In addition, multivariate regression analysis in the original cohort and propensity score-matched Cox analysis demonstrated that receiving HBcA b+ liver grafts was not a significant risk factor for long-term survival. These findings were consistent in both HBV surface antigen-positive(HBsAg+) and HBsA g negative(HBsAg-) patients.Newly diagnosed HBV infection had a relatively higher incidence in HBsAg-patients with HBcAb+ liver grafts(13.23%), in which HBV naive recipients suffered most(31.82%), although this difference did not affect patient and graft survival(P = 0.50 and P = 0.49, respectively). Recipients with a high HBV surface antibody(anti-HBs) titer(more than 100 IU/L) before transplantation and antiviral prophylaxis with nucleos(t)ide antiviral agents post-operation, such as nucleos(t)ide antiviral agents, had lower de novo HBV infection risks. CONCLUSION HBcA b+ liver grafts do not affect the long-term outcome of the recipients. Combined with proper postoperative antiviral prophylaxis, utilization of HBcAb+ grafts is rational and feasible.

Association of core promoter mutations of hepatitis B virus and viral load is different in HBeAg(+) and HBeAg(-) patients

AIM:To identify the prevalence of hepatitis B e antigen (HBeAg) and to assess the association of hepatitis B virus (HBV) core promoter mutations and viral load in Indonesian patients.METHODS:Sixty-four patients with chronic hepatitis,65 with liver cirrhosis and 50 with hepatocellular carcinoma were included in this study.HBeAg and hepatitis B e antibody (HBeAb) tests were performed using enzyme-linked immunosorbent assay and the mutations were analyzed by sequencing.Viral load was measured by real-time polymerase chain reaction.RESULTS:Of 179 patients,108 (60.3%) were HBeAg(-) and 86 (79.6%) of these HBeAg(-) patients had been seroconverted.The A1896 mutation was not found in HBeAg(+) patients,however,this mutation was detected in 70.7% of HBeAg(-) patients.This mutation was frequently found when HBeAg was not expressed (87.7%),compared to that found in HBeAg seroconverted patients (65.1%).The A1899 mutation was also more prevalent in HBeAg(-) than in HBeAg(+) patients (P=0.004).The T1762/A1764 mutation was frequently found in both HBeAg(+) and HBeAg(-) patients,however,the prevalence of this mutation did not significantly differ among the two groups (P=0.054).In HBeAg(+) patients,the T1762/A1764 mutation was correlated with lower HBV DNA (P < 0.001).The A1899 mutation did not correlate with HBV DNA (P=0.609).In HBeAg(-) patients,the T1762/A1764 mutation alone was not correlated with HBV DNA (P=0.095),however,the presence of either the T1762/A1764 or A1896 mutations was associated with increased HBV DNA (P < 0.001).CONCLUSION:The percentage of HBeAg(-) patients is high in Indonesia,and most of the HBeAg(-) patients had been seroconverted.The A1896 mutation was most likely the major cause of HBeAg loss.The T1762/A1764 mutation alone was associated with lower viral loads in HBeAg(+) patients,but not in HBeAg(-) patients.

Hepatitis B virus reactivation in hepatitis B virus surface antigen negative patients receiving immunosuppression: A hidden threat

AIM: To present the characteristics and the course of a series of anti- hepatitis B virus core antibody (HBc) antibody positive patients, who experienced hepatitis B virus (HBV) reactivation after immunosuppression. METHODS: We retrospectively evaluated in our tertiary centers the medical records of hepatitis B virus surface antigen (HBsAg) negative patients who suffered from HBV reactivation after chemotherapy or immunosuppression during a 3-year period (2009-2011). Accordingly, the clinical, laboratory and virological characteristics of 10 anti-HBc (+) anti-HBs (-)/HBsAg (-) and 4 anti-HBc (+)/antiHBs (+)/HBsAg (-) patients, who developed HBV reactivation after the initiation of chemotherapy or immunosuppressive treatment were analyzed. Quantitative determination of HBV DNA during reactivation was performed in all cases by a quantitative real time polymerase chain reaction kit (COBAS Taqman HBV Test; cut-off of detection: 6 IU/mL). RESULTS: Twelve out of 14 patients were males; median age 74.5 years. In 71.4% of them the primary diagnosis was hematologic malignancy; 78.6% had received rituximab (R) as part of the immunosuppressive regimen. The median time from last chemotherapy schedule till HBV reactivation for 10 out of 11 patients who received R was 3 (range 2-17) mo. Three patients (21.4%) deteriorated, manifesting ascites and hepatic encephalopathy and 2 (14.3%) of them died due to liver failure. CONCLUSION: HBsAg-negative anti-HBc antibody positive patients can develop HBV reactivation even 2 years after stopping immunosuppression, whereas prompt antiviral treatment on diagnosis of reactivation can be lifesaving.

Occult hepatitis B virus infection and blood transfusion

Transfusion-transmitted infections including hepatitis B virus(HBV) have been a major concern in transfusion medicine. Implementation of HBV nucleic acid testing(NAT) has revealed occult HBV infection(OBI) in blood donors. In the mid-1980 s, hepatitis B core antibody(HBc) testing was introduced to screen blood donors in HBV non-endemic countries to prevent transmission of non-A and non-B hepatitis. That test remains in use for preventing of potential transmission of HBV from hepatitis B surface antigen(HBs Ag)-negative blood donors, even though anti-hepatitis C virus testshave been introduced. Studies of anti-HBc-positive donors have revealed an HBV DNA positivity rate of 0%-15%. As of 2012, 30 countries have implemented HBV NAT. The prevalence of OBI in blood donors was estimated to be 8.55 per 1 million donations, according to a 2008 international survey. OBI is transmissible by blood transfusion. The clinical outcome of occult HBV transmission primarily depends on recipient immune status and the number of HBV DNA copies present in the blood products. The presence of donor anti-HBs reduces the risk of HBV infection by approximately five-fold. The risk of HBV transmission may be lower in endemic areas than in non-endemic areas, because most recipients have already been exposed to HBV. Blood safety for HBV, including OBI, has substantially improved, but the possibility for OBI transmission remains.

“Anti-HBc alone” in human immunodefi ciency virus-positive and immuno-suppressed lymphoma patients

Hepatitis B virus (HBV) infection is endemic in various parts of the world. A proportion of patients have resolved prior exposure to HBV, as evidenced by the clearance of circulating hepatitis B surface antigen and the appearance of antibody to hepatitis B core antigen (anti-HBc), which could produce protective antibody to hepatitis B surface antigen (anti-HBs). With time, anti-HBs in some patients may become negative. Such patients are described as having occult HBV infection or "anti-HBc alone". In the context of immunodef icient patients, such as HIV patients or lymphoma patients undergoing immunosuppressive immunotherapy, the lack of protective anti-HBs may increase the risk of hepatitis B reactivation. Serum HBV DNA testing may be necessary in "anti-HBc alone" patients, to detect patients at a high risk of developing HBV infection allowing appropriate prophylactic management.

Establishment of transgenic mouse harboring hepatitis B virus (adr subtype) genomes

INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For example, a statistical data from ahospital in Shanghai showed that 80% of HCCpatients were positive for HBsAg ( personalcommunication).

干扰素抗体与HBV C基因启动子变异对干扰素疗效的影响

目的 探讨HBVC基因启动子 (BCP)变异的肝炎患者干扰素抗体 (抗 IFN )的产生对干扰素疗效的影响。方法 采用错配PCR RFLP技术检测 89例慢性乙型肝炎血清中BCP变异和血清中抗 IFN的水平。结果 BCP变异率为 5 2 .8%,抗 IFN阳性率为 15 .6 %,而干扰素治疗前抗 IFN阳性与治疗后转为阳性的患者 ,其干扰素治疗的有效率差异无显著性 (P >0 .0 5 ) ,而治疗前后抗 IFN均为阴性者 ,其有效率明显优于阳性者 ,差异显著 (P <0 .0 5 )。在IFN治疗前 ,有BCP变异组抗 IFN阳性 10例 ,野生株组抗 IFN阳性 2例 ,两组相比 ,差异显著 (P <0 .0 5 )。IFN治疗后 ,变异株组出现抗 IFN阳性 4例 ,野生株组出现抗 IFN阳性 1例 ,差异无显著性 (P >0 .0 5 )。结论 抗 IFN阳性的患者 ,干扰素治疗效果差。BCP变异的患者 ,在干扰素治疗前 ,有可能促进干扰素抗体的产生 ,但在干扰素治疗后 ,则不增加抗 IFN的产生。

HBcAb阳性伴HBsAb阴性的人群HBsAg合适临界值的研究

目的:HBcAb阳性伴HBsAb阴性的人群HBsAg合适临界值的研究。方法:收集并检测HBcAb阳性伴HBsAb阴性组HBsAg阴性及HBcAb阴性伴HBsAb阴性组HBsAg阴性的样本各1 522例,比较两组HBsAg结果的差异。采用ROC曲线对ELISA定性HBsAg结果诊断HBcAb阳性伴HBsAb阴性的人群的阳性判断值的再确定,寻找更好的的诊断性能。结果:i2000SR检测HBsAg的结果具有良好的重复性,符合临床检测要求。HBcAb阴性组复检前与复检后相比明显降低,复检后HBcAb阳性组的结果相比阴性组明显降低,差异具有统计学意义(P<0.05);利用ROC曲线对HBcAb阳性伴HBsAb阴性的人群HBsAg建立合适临界值为0.415后的准确度、敏感度、特异度、阳性预测值、阴性预测值、曲线下面积、Youden指数显示具有更好的诊断性能。HBcAb阳性伴HBsAb阴性人群的HBsAg结果与HBcAb阴性伴HBsAb阴性人群HBsAg的相比明显升高,差异具有统计学意义(P<0.05)。结论:实验室建立HBcAb阳性伴HBsAb阴性人群的ELISA法定性HBsAg结果合适的的阳性判值具有更好的诊断性能。

儿童与成人慢性乙型肝炎患者乙型肝炎病毒Core基因区准种特征及正选择压力差异分析

儿童与成人慢性乙型肝炎患者的临床特征差异明显。乙型肝炎病毒(Hepatitis B virus,HBV)病毒准种特征与其致病特性紧密相连,HBV病毒Core基因区富含免疫表位,该区域的准种特征直接反映病毒变异与病毒应对宿主免疫压力间的动态过程。文章通过扩增170名儿童慢性乙型肝炎患者及121名成人慢性乙型肝炎患者病毒Core基因区,按照病毒基因型以及病毒e抗原(Hepatitis B virus e antigen,HBe Ag)状态进行分组,使用序列复杂度、多样性、非同义突变率(Non-synonymous substitution ratio,d N)、同义突变率(Synonymous substitution ratios,d S)等指标衡量不同组别之间的病毒准种特征;使用不同模型计算不同组别中受到正选择压力的位点,进一步结合HBV Core基因区免疫表位信息,进行正选择位点的定位分析。结果发现,儿童乙型肝炎病毒患者体内病毒Core基因区序列复杂性和多样性低于成人患者,且前者Core基因区正选择位点个数显著低于后者,这说明儿童慢性乙型肝炎患者体内病毒受到的选择压力低于成人患者。在儿童及成人慢性感染病人组中,HBe Ag阳性病人体内病毒受到的选择压力低于HBe Ag阴性病人。儿童及成人慢性感染患者体内病毒存在13个正选择位点,大多数正选择位点位于已知的抗原表位上。本研究从分子进化角度揭示了儿童与成人慢性乙型肝炎病例体内病毒Core基因区序列准种差异,为两类病人显著不同的临床表征提供了群体遗传学的解释。

抗鸭乙肝病毒core蛋白单克隆抗体的制备与鉴定

目的制备抗鸭乙肝病毒core蛋白的单克隆抗体并进行鉴定。方法 PCR扩增鸭乙肝病毒(DHBV)core基因片段,构建表达载体pET28a(+)/DHBV core。转化诱导表达融合蛋白,用Ni 2+亲和柱纯化目的蛋白。免疫Balb/c小鼠,取脾细胞与SP2/0骨髓瘤细胞融合,ELISA筛选,采用有限稀释法筛选单克隆杂交瘤细胞并进行细胞克隆。体内诱生法大量制备单克隆抗体,进行抗体的特异性、效价和亚型的鉴定。结果成功构建表达载体pET28a(+)/DHBV core,并表达DHBV core蛋白。获得2株稳定分泌抗DHBV core抗体的杂交瘤细胞株,分别为2D7和5G10,细胞培养液抗体效价为1∶400和1∶800。选择5G10细胞株制备单克隆抗体,小鼠腹水抗体效价可达1∶320 000。鸭肝组织免疫组化结果显示,DHBV病毒载量>1010copies/mg的肝组织中DHBV core蛋白的表达明显高于病毒载量<105copies/mg的肝组织,而未感染DHBV的鸭肝细胞内不表达DHBV core蛋白。亚型鉴定结果为IgG2aκ链。结论制备并获得了抗DHBV core蛋白的单克隆抗体,为DHBV core蛋白的功能研究、诊断试剂的研制与抗病毒治疗研究奠定了基础。

乙肝患者血清抗原与抗体双阳性两对半模式与HBV-DNA和体液免疫检测结果分析

目的：探讨乙型肝炎病毒（HBV）感染者HBsAg阳性的血清学标志物少见模式与HBV-DNA和体液免疫的关系。方法：选择63例HBsAg阳性患者其血清中检出同一种抗原抗体同时存在（HBsAg与HBsAb同时阳性或 HBeAg与HBeAb同时阳性），以HBeAg是否阳性为标准分为2组：HBeAg阳性组（n=26）与HBeAg阴性组（n=37）。采用时间分辨免疫荧光分析法（TRFIA）定量测定乙型肝炎病毒血清标志物（HBV-M）；采用荧光定量聚合酶链反应（FQ-PCR）法检测HBV-DNA含量；采用Olympus AU6400全自动生化分析仪检测免疫球蛋白（IgG、IgA 、IgM）和补体（C3、C4）。结果：63例HBsAg阳性的血清学标志物少见模式表现为5种模式，以HBsAg（＋） HBsAb（＋）HBeAb（＋）HBcAb（＋）模式检出率最高，占52.38%（33/63）；各少见模式均不同程度地检出HBV-DNA；HBeAg阳性组HBV-DNA、IgG、IgM和C4与HBeAg阴性组比较差异有统计学意义（P〈0.05）；IgA和C3的差异均无统计学意义（P〈0.05）。结论：患者在慢性HBV感染过程中，血清出现抗原与抗体共存这一个特殊阶段，虽然有HBsAb的出现，但并不意味着HBV-DNA停止复制或传染性消失；不同血清学模式，其HBV复制和体液免疫水平是有差别的。

灰旱獭感染HBV的临床和病理分析

目的 探讨灰旱獭感染HBV后临床表现、病理改变及其相关关系。方法 ①用含HBV DNA人血清对健康灰旱獭攻毒 ,依次传代 ,分析其临床血清学变化与病理改变的相关性。②用合成HBcAg多肽分别接种 :对HBcAb- 旱獭单用多肽Ⅰ、Ⅱ或多肽Ⅰ、Ⅱ与HBV DNA同时注射 :对HBcAb+ 旱獭注射多肽Ⅰ、Ⅱ。结果 ①攻毒与传代后临床反应的轻重与肝脏病理变化的程度一致。②HBcAg多肽具有核心蛋白疫苗的作用。可激发抗体的形成 ,阻断HBV的攻毒 ,使HBcAb转阴 ,与肝组织炎症反应一致。结论 肝组织炎变与血清变化密切相关 ,证实攻毒、传代结果可靠。多肽具有阻断、干预灰旱獭感染HBV的作用。

抗HBcAg人源性单链抗体细胞内表达及其抗病毒复制的实验研究

应用人源性抗HBcAg单链抗体细胞内表达技术 ,探讨抗HBV复制基因治疗的应用价值。应用噬菌体展示和基因重组技术 ,从HBV感染的外周血淋巴细胞克隆了人源性抗HBcAg单链抗体 ,并重组至逆转录病毒载体。以人肝癌细胞smmc - 772 1和PLC/PRF/5为靶细胞进行基因共转染 ,分别测定实验组细胞上清中的HBsAg和HBeAg ,与对照组做比较 ,观察抗HBcAg单链抗体细胞内表达的抗病毒治疗作用。结果显示 ,在急性HBV感染的细胞株中 ,抑制病毒复制效率为 4 9%～ 6 1% ,在慢性病毒感染细胞 ,抑制率为 4 1%～ 5 4 %。实验结果表明 ,应用单链抗体细胞内表达技术 ,在抗病毒治疗研究中具有潜在的应用价值。应对HBV的 4个开放阅读框架编码产物进行全面的对比研究 ,以发现抑制效率高。

HBsAg阴性/抗HBc阳性献血员HBV感染状况调查

目的 对431例HBsAg阴性献血员检测抗-HBc、抗-HBs,抗-HBc阳性者196例(45.5％),其中单项抗-HBc阳性者35例(17.9％),抗-HBc/抗-HBs阳性者161例(82.1％)。对抗-HBc阳性者检测抗-HBc IgM和HBV DNA(聚合酶链法),抗-HBc IgM的检出率为32.1％(63/196),其中抗-HBc/抗-HBs阳性者检出率为29.8％(48/161),单项抗-HBc阳性者检出率为42.9％(13/35),二者无差异;HBV DNA检出率为14.8％(29/196),单项抗-HBc阳性者HBV DNA检出率为25.7％,显著高于抗-HBs/抗-HBc阳性者(12.4％)(P<0.05);抗-HBc IgM阳性者HBV DNA检出率(39.7％)也显著高于抗-HBc IgM阴性者(3.0％)(P<0.001),二者阴阳性符合率则为78.6％(154/196)。HBsAg阴性/抗-HBc阳性献血员仍有传染性存在,尤以抗-HBc IgM阳性者最具血源传播HBV的危险性,因此,建议对HBsAg阴性献血员再进一步筛检抗-HBc IgM。

化学发光免疫分析法检测HBcAb阳性、HBsAg阴性血清中隐匿性乙型肝炎病毒感染的分析

目的分析化学发光免疫分析法(CIA)检测乙型肝炎病毒核心抗体(HBcAb)阳性、乙型肝炎病毒表面抗原(HBsAg)阴性血清学模式中隐匿性乙型肝炎病毒感染(OBI)的情况,旨在了解CIA检测HBsAg的分析性能,并为评价OBI提供实验室依据。方法收集2017年5月至2017年12月本院检验科近20000份CIA检测乙型肝炎病毒(HBV)五项血清学标志物中HBsAg无反应性、HBcAb阳性且信号值比临界信号值(S/Co值)>7.5的样本1157例,对样本HBcAb再进行酶联免疫吸附试验(ELISA)复筛,选取A450nm吸光值<0.070样本294例进行HBV DNA检测。结果 1157例样本进行HBcAb ELISA检测,复筛得HBcAb阳性标本1053例,阴性104例,其中A450nm吸光值<0.070样本294例。294例样本进行HBV DNA检测,其中HBV DNA阳性(HBV DNA>20IU/mL为阳性)34例,总阳性率为2.94%(34/1157)。22例HBV DNA载量21～99IU/mL;7例为100～1000IU/mL;5例为> 1000IU/mL;主要以低水平形式存在。以HBeAb、HBcAb阳性及HBsAb、HBeAb和HBcAb阳性这两种血清学模式中HBV DNA检出率较高;HBcAb S/Co≥10.0样本HBV DNA阳性率可达13. 5%。结论 CIA作为检测HBsAg灵敏度较高的方法 ,依然存在HBsAg阴性而HBV DNA阳性的OBI。这对于输血的安全性和乙肝的预防监测及治疗都存在重大影响。临床上对于HBsAg阴性伴高反应性HBcAb(S/Co值≥10.0)的患者应进行一步行HBV DNA检测,以明确OBI的存在。

抗-HBc定量测定在乙型肝炎病毒慢性感染性疾病中的应用

乙肝病毒核心抗体(抗-HBc)与机体免疫相关,在现症及既往乙型肝炎病毒(HBV)感染者血清中均可测得,其在HBV自然史的判断及治疗策略中可发挥巨大作用。HBV感染慢性化与肝内共价闭合环状DNA(cccDNA)活跃转录和持续存在密切相关;抗-HBc定量与cccDNA水平有较好相关性,可作为综合评价HBV感染性疾病的一项指标。

病毒性肝炎患者柯萨奇B组病毒感染状况的研究

目的研究病毒性肝炎患者的柯萨奇B组病毒感染状况。方法用免疫酶组化法对121例各型临床肝炎患者和108名献血员进行了抗-CVBIgM和IgG检测。结果两组阳性率分别为9．9％、19．8％和8．3％、36．1％，总阳性率分别为25．6％、40．7％；非乙型肝炎患者抗-CVBIgM、IgG及总阳性率为13．8％、24．1％和31．0％，乙肝患者抗-CVBIgM、IgG及总阳性率分别为6．3％、15．9％和20．6％。肝炎患者，尤其是乙肝患者抗-CVBIgG及总阳性率与对照组差异有显著性。结论乙肝患者可能对CVB感染的免疫应答较低。

抗重组HBcAg单克隆抗体的建立和应用

用硫酸铵沉淀方法纯化的重组HBcAg(rHBcAg作为免疫原免疫小鼠,获得 5株高滴度和特异性的HBc单克隆抗体(rHBcMcAb),纯化并酶标后与rHBcAg组合后用于血清HBcAb检测的试剂盒。经实验证明,该单克隆抗体具有特异性强、敏感和重复性好的优点,经不同株间的单克隆组合应用效果较好,可以代替具有污染可能性的血清来源的抗体。同时也证明经过初步纯化的rHBcAg为较好的免疫原。

Isolated anti-HBc is an independent risk factor for tumor recurrence in intrahepatic cholangiocarcinoma after curative resection

Background:Intrahepatic cholangiocarcinoma(ICC)is a poorly understood and aggressive malignancy with increasing incidence and mortality.Hepatitis B virus(HBV)infection is recognized as one of the important risk factors of ICC.There are few reports focusing on whether isolated antibody to hepatitis B core antigen(isolated anti-HBc,IAHBc)have prognostic role in ICC,while positive hepatitis B surface antigen(HBsAg)has been reported to be associated with the prognosis of ICC.The aim of this study was to investigate the prognostic value of IAHBc in ICC patients after curative resection,in order to identify those who have the high risk of ICC recurrence in the early stage.Methods:We divided 209 ICC patients who underwent curative resection into 4 groups:groupⅠ(n=40),HBsAg(-)/antibody to hepatitis B surface antigen(anti-HBs)(-)/anti-HBc(+);groupⅡ(n=70),HBsAg(+)/anti-HBc(-);groupⅢ(n=55),HBsAg(-)/anti-HBs(+)/anti-HBc(+);and groupⅣ(n=44),HBsAg(-)/anti-HBc(-).We compared the recurrence-free survival(RFS)and overall survival(OS)among these four groups.Results:The median follow-up time was 16.93 months(range 1-34.6 months).The 1-and 2-year RFS and OS rates were 60%and 42%,and 78%and 63%respectively in all patients.Compared to the whole non-IAHBc patients(groupⅡ+groupⅢ+groupⅣ),IAHBc patients(groupⅠ)showed significantly lower RFS at 1 year(39.8%vs.64.4%,P=0.001)and 2 years(20.7%vs.46.7%,P=0.001).When compared to other three individual groups,IAHBc patients(groupⅠ)also had the lowest RFS.We did not find significant difference in OS among the four groups.Further multivariate analysis revealed that IAHBc was an independent risk factor of RFS.Conclusions:IAHBc is an independent poor prognostic factor for tumor recurrence in ICC patients after curative resection.

HBV核心蛋白和B7.1分子嵌合质粒的构建及体外表达的初步研究

目的 构建HBV核心蛋白和B7.1分子真核表达嵌合质粒 ,并检测其在体外的表达。方法 利用亚克隆技术 ,将HBV核心基因片段和B7.1基因片段构建至真核表达质粒 pcDNA3中 ,测定序列后 ,用脂质体包裹转染细胞 ,采用间接免疫荧光法检测两种蛋白的表达。结果 核酸序列测定证实实验所构建的质粒正确 ,嵌合质粒中所含的HBcAg和B7.1分子的核苷酸序列与HBVadr亚型标准株核心区及鼠B7.1分子的同源性分别为 98.18%和 99.89%。该嵌合质粒在体外转染细胞内可表达HBcAg和B7.1分子。 结论 实验所构建的HBcAg/B7.1嵌合质粒能在体外同时表达HBcAg和B7.1分子。