HBV C基因型有关的HBsAg阴性HBV DNA阳性患者S区突变对HBsAg的影响

目的通过构建HBV C基因型突变质粒研究HBsAg阴性HBV DNA阳性患者HBV S区突变对HBsAg水平的影响。方法收集2022年8月至2023年4月首都医科大学附属北京佑安医院107例HBsAg-/HBV DNA+患者血液样本,对成功提取扩增的HBV DNA S区进行测序,通过构建HBV C基因型突变质粒对HBV S区突变位点进行细胞功能验证,探讨OBI可能发生的分子机制。结果对成功提取扩增的68例患者进行测序,发现HBV S区存在大量突变,包括免疫逃逸突变(如sG145R、sK122R、sS114T、sT131P等)和跨膜结构域(transmembrane domain,TMD)突变(如sT5A、sG10D、sF20S等)。通过构建HBV C基因型突变质粒,进行细胞转染和细胞免疫荧光实验发现sG145R突变会明显降低HBsAg的表达,但是sK122R、sI26N、sQ29N、sR169H、sS114T、sT131P这6个突变位点并未影响细胞内外HBsAg的表达。结论通过测序发现HBsAg-/HBV DNA+患者HBV S区存在大量突变位点,通过构建sG145R、sK122R、sI26N、sQ29N、sS114T和ST131P等突变质粒发现sG145R突变会明显降低细胞内外HBsAg的表达,但是sK122R、sI26N、sQ29N、sR169H、sS114T、sT131P并未明显降低细胞内外HBsAg的表达。

Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA

AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs).METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19×1012copies of linear HBV DNA/1×107 PDHs). After 1-5 d of transfection, HBsAg and HBeAg in the supernatant and lysate of PDHs were measured with the IMX System.Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PDHs electroporated were used as control group.RESULTS: HBsAg in the hepatocyte lysates of transfected group was 15.24 (1 d), 14.55 (3 d) and 5.13 (5 d; P/N values, positive≥2.1) respectively. HBeAg was negative (＜2.1). Both HBsAg and HBeAg were negative in the supernatant of transfected group. Dot blotting revealed that HBV DNA was strongly positive in the transfected group and negative in the control group. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (rc DNA), covalently closed circular (ccc DNA), and single-stranded (ss DNA) HBV DNA replicative intermediates in the transfected group, there was no integrated HBV DNA in the cellular genome. These parameters were negative in control group.CONCLUSION: Expression and replication of HBV genes can occur in hepatocytes from non-mammalian species.HBV replication has no critical species-specificity, and yet hepatic-specific regulating factors in hepatocytes may be essential for viral replication.

The Effect of Gankang Suppository on Duck Hepatitis B Virus, Serum Biochemistry and Liver Histology in Ducklings

To examine the effect of Gankang Suppository on duck hepatitis B virus （DHBV）, the serum biochemistry and hepatic histology in an animal model of DHBV infection, a model of DHBV infection was established by infecting 1-day-old Yingtaogu ducklings with DHBV-positive serum. The successful model was confirmed by PCR assay and 48 ducklings infected with DHBV were randomly divided into 3 groups： a Gankang Suppository treatment group, an acyclovir （ACV） group and a DHBV model group （control）, with each group having 16 animals. All the animals were given the medicines for 4 weeks in a row. The serum of the animals was taken 14 and 28 days after the medica- tion and 7 days after drug discontinuation. Real-time PCR was performed to detect the copy numbers of DHBV DNA in the serum. ALT and AST were dynamically monitored. The ducklings were sacrificed on the 7th day after the discontinuation of the treatment and livers were harvested and examined for inflammation and degeneration of liver cells by using HE staining. The results showed that on day 14, 28 after the treatment and day 7 after the withdrawal, the logarithmic values （log） of DHBV DNA copy numbers in ducklings of Gankang Suppository treatment group were significantly lower than that before the treatment （P=0.0092, P=0.0070, P=0.0080, respectively）. Compared with DHBV model control group, the ALT level was significantly decreased （P=0.0020, P=0.0019, respectively） on day 28 after the treatment and on day 7 after the withdrawal. The AST level was also reduced on day 14 after the treatment （P=0.0298）. Compared with the ACV control group, the level of ALT was lower on day 7 after the withdrawal （P=0.0016）. Histologically, the hepatocyte swelling, vacuolous degeneration and acidophilic degeneration in Gankang Suppository treatment group were alleviated 7 days after the withdrawal as compared with model control group （P=0.0282, P=0.0084, P=0.0195, respectively）. It is concluded that Gankang Suppository can effectively suppress DHBV replication, reduce the levels of serum ALT and AST and improve hepatic histology.

The Potential of Duck Hepatitis Virus(DHV-1) Stimulating the Body Weight Gain and the Effects of Silymarin on It in Duckling

To evaluate the effects of duck hepatitis virus-1 （DHV-1） on the body weight gain in duck and the effects of silymarin on it in vivo, 100 10-d-old ducks, both male and female, were collected to be subjected to the test. The experiments were conducted in 8 groups： in group 1-3, the animals were inoculated with 1：105 diluted duck hepatitis virus （DHV-1） infected allantoic fluid and given 0, 30, and 50 mg kg^-1 BW d^-1 silymarin orally, respectively. In group 4-6, the animals were inoculated with 1：5 × 105 diluted DHV-1 infected allantoic fluid and given 0, 10, and 30 mg kg^-1 BW d^-1 silymarin orally, respectively. In group 7, the animals were given 10 mg kg^-1 BW d^-1 silymafin only. Group 8 was the control one treated by injecting sterillized saline into the leg muscles. All the silymarin was given from 0 to 4 d after inoculation of the virus. By the 5th d after inoculation, the vein blood was drawn from the dorsal foot vein and the plasma samples were collected and stored at -20℃. The body weight gain （BWG） was measured from 0 to 10 d after inoculation. The plasma IGF-I, T3, and T4 concentrations were measured by radioimmunoassay （RIA）. At the virus dose of 1：5 ×105 diluted virus infected allantoic fluid, the inoculation of the virus enhanced the BWG significantly compared with that of the control （P〈 0.01）, while 10-50 mg kg^-1 BW d^-1 silymarin could counteract the effects of the virus on the BWG dose-dependently. The plasma IGF-I levels showed no correlation with the BWG, but the T3 levels showed a same tropism with the body weight gain. The present results indicated that sublethal DHV-1 enhanced the body weight gain of ducklings significantly, and the silymarin could counteract this effect in vivo.

Molecular Characterization of Duck Hepatitis B Virus Isolated from Hubei Brown Ducks

The objective of this study was to characterize the genome structure of duck hepatitis B virus （DHBV） isolated from Hubei brown ducks. The natural carrier rate of DHBV in adult ducks from Hubei area was investigated and the DHBV DNA-positive serum screened out. The complete genome of a DHBV strain was amplified by polymerase chain reaction （PCR） and cloned into T vector and sequenced. The results showed that the carrier rate of DHBV in Hubei brown ducks was 10 % This strain （GenBank accession number DQ276978） had a genome of 3024 nucleotides with three overlapping open reading frames encoding the surface, core and polymerase proteins respectively. Comparison of the strain with 17 DHBV strains registered in GenBank revealed a homology from 89.3 % to 93.5 % at the nucleotide level. The sequences of the structural and functional domains of these proteins were highly conserved. The strain was found to share more signature amino acids in the polymerase genes with the ＂Chinese＂ DHBV strains than those of the ＂Western＂ country strains. This finding was also corroborated by a phylogenetic tree analysis. Therefore, the DQ276978 might belong to a subtype of the Chinese DHBV strains.

Therapeutic effect of Styela plicata on duck hepatitis B virus in vivo

Objective:To evaluate the antiviral activity of the alcohol extract of Styela plicata on DHBV (duck hepatitis B virus) in vivo. Methods: Guangzhou-Sheldrake ducklings congenitally infected with DHBV were assigned to receive the alcohol extract of Styela plicata or lamivudine for 30 consecutive days. The DHBV DNA of sera was detected by RT-PCR. and the histological analysis of duckling liver was evaluated. Results:Thirty days after therapy,histological analysis of duckling liver showed that the ducklings receiving the alcohol extract of Styela plicata or lamivudine exhibited catabatic status in the degree of liver cell degeneration and inflammation compared with the ducklings receiving normal diet. DHBV DNA of sera from alcohol extract of Styela plicata-treated ducklings and lamivudine-treated ducklings all produced significantly lower levels compared with ducklings receiving normal diet (P<0. 01 ). Although these treatment groups all exhibited a rebound phenomenon 10 d after withdrawal of medication, they still exhibited a significant lower level of serum DHBV DNA compared with the control group responded to normal diet (P<0. 05, P<0. 01). Conclusion:Styela plicata may be an effective antiviral medicine in treating chronic hepatitis B. The data of this experiment will be valuable in studying the therapeutic role and the potential therapeutic mechanism of Styela plicata.

DUCK HEPATITIS B VIRUS DNA WITHIN HEPATIC MULTICENTRIC CANCER AND/OR METASTATIC CANCER

Duck hepatitis B vims (DHBV) DNA was detected in different tumorous nodules of ducks with hepatic multicentric cancer or intrahepatic metastasis by Southern blot technique. Among 7 ducks with hepatocellular carcinoma of multiple tumor nodules, the hybridization pattern of Integrated DHBV DNA In different tumorous nodules was identical in 3 cases and different in 2 cases. One case showed a similar hybridization pattern in two tumorous nodules and other one was negative tor DHBV DNA. Integrated DHBV DNA was also identified in a metastatic lung cancer of ducks with hepatocellular carcinoma. The hybridization pattern of metastasis of lungs was as the some as that in primary hepatocellular carcinoma. The same discrete hybridization bands In the different tumorous nodules indicate that these nodules might arise from one transformed cell. The different hybridization patterns In various tumorous nodules show that these tumorous nodules might arise from various transformed cells. The results suggest that the hybridization pattern of different nodules of hepatocellular carcinoma with viral DNA probe could make a cell clone origin marker of tumor nodule to differentiate hepatic multlcentric cancer from Intrahepatic metastatic cancer.

Detection of the covalently closed circular DNA of duck hepatitis B virus by Taq-Man fluorescent quantitative PCR assay

To develop a fluorescent quantitative PCR assay based on Taq-Man chemistry to detect the covalenfly closed circular DNA （eccDNA） of duck hepatitis B virus （DHBV）, a pair of primers was designed from both sides of the nick in the minus strand of DHBV and a Taq-Man probes between the primers, modified with 6-Fam at 5＇ end and Tamra at its 3＇ end was designed to detect the PCR products during PCR cycles. The DHBV DNA fragment was cloned into vector PUCm-T, and the recombinant plasmid was purified and subsequently qualified as the HBV DNA standard. The experimental conditions and reagents used in PCR assay for amplification were sophisticatedly optimized in order to yield a perfect amplification efficacy and reduce the possibility to produce non-specific amplification. It was demonstrated that the detect limit of assay was 10^3 copies/ml, and a linear standard curve was obtained between 10^5 -10^9 copies/ml [ C1 =-2.8361 ln（x） ＋ 41.45, r =-0.9985]. The coefficient of variation was 0.2%-3.14% and 2.22%-4.43% for intra- and inter-assay respectively. After a dynamic survey on the contents of DHBV DNA in serum of ducks, it was found that its peak value appeared at the second week of birth in ducks. It is evident that this method of Taq-Man fluorescent quantitative PCR assay appears to be simple, sensitive and specific.

HBV感染相关性肝细胞癌患者血甲胎蛋白、转氨酶水平与HBV-DNA载量和病情进展的关系

目的探讨乙型肝炎病毒(HBV)感染相关性肝细胞癌(HCC)患者血甲胎蛋白(AFP)、转氨酶水平与HBV-DNA载量和病情进展的关系。方法回顾性分析2022年1月至2023年4月本院收治的50例HBV感染相关性HCC患者资料并设为HCC组,另筛选同期HBV相关肝硬化(LC)和慢性乙型肝炎(CHB)患者资料各30例设为LC组和CHB组,比较三组血AFP及转氨酶水平;比较HCC组不同HBV-DNA载量患者AFP及转氨酶水平差异,并分析肝癌组患者AFP及转氨酶水平与病情进展的关系。结果HCC组患者AFP和丙氨酸氨基转移酶(ALT)水平均高于CHB组和LC组(P<0.05);50例HCC患者中,中低分化及Ⅲ~Ⅳ期患者血AFP和ALT水平高于高分化及Ⅰ~Ⅱ期患者(P<0.05);HBV-DNA高载量患者血AFP和ALT水平高于低载量患者(P<0.05),且患者HBV-DNA载量与其血AFP和ALT水平呈正相关(P<0.05)。结论HBV感染相关性HCC患者血AFP和ALT水平显著提升,且血AFP和转氨酶水平与HBV-DNA载量、肿瘤分期分化程度关系密切,二者对临床评估HBV感染相关性HCC病情进展有一定提示作用。

合肥地区献血者血液筛查非重复反应性标本与OBI标本间HBV血清学特征分析

目的 了解血液筛查非重复反应性标本与隐匿性乙型肝炎病毒感染(OBI)标本间HBV血清学特征的相关性。方法 收集本科室2021年1月—2023年1月血液筛查ELISA结果均为阴性,仅NAT反应性的标本共144份,其中TMA法联检反应性标本92份,PCR法单人份检测HBV DNA反应性标本52份。联检反应性标本补充TMA法鉴别检测和PCR法单人份检测,2种方法检测均无反应性的标本纳入NRR标本组,任1种方法检出HBV DNA反应性的标本纳入OBI标本组。对2组标本完成HBsAg、抗-HBs、HBeAg、抗-HBe、抗-HBc检测,分析NRR标本与OBI标本在血清学模式和阳性率上是否存在差异。结果 联检反应性标本补充检测均阴性标本53份,纳入NRR标本组。91份标本在任1种方法中检测出HBV DNA反应性,纳入OBI标本组。2组标本血清学检测均未检出HBsAg和HBeAg,抗-HBs检出率NRR标本组为64.15%,OBI标本组为47.25%;抗-HBc检出率NRR标本组为86.79%,OBI标本组为94.51%;抗-HBe检出率NRR标本组为35.85%,OBI标本组为52.75%。2组标本血清学模式:NRR标本组表现最多的模式为抗-HBs+、抗-HBc+(32.08%),OBI标本组表现最多的模式为抗-HBe+、抗-HBc+(37.36%)。结论 NRR标本与OBI标本在HBV血清学检测中的部分检测结果间存在差异,但NRR标本中较高的抗-HBc阳性率提示仍有较高的可能存在HBV感染风险。

Polymorphisms of microsomal triglyceride transfer protein in different hepatitis B virus-infected patients

AIM: To identify the two polymorphisms of microsomal triglyceride transfer protein (MTP) gene in the Chinese population and to explore their correlation with both hepatitis B virus (HBV) self-limited infection and persistent infection. METHODS: A total of 316 subjects with self-limited HBV infection and 316 patients with persistent HBV infection (195 subjects without familial history), matched with age and sex, from the Chinese Han population were enrolled in this study. Polymorphisms of MTP at the promoter region -493 and at H297Q were determined by the allele specific polymerase chain reaction (PCR). RESULTS: The ratio of males to females was 2.13:1 for each group and the average age in the self-limited and chronic infection groups was 38.36 and 38.28 years, respectively. None of the allelic distributions deviated significantly from that predicted by the Hardy-Weinberg equilibrium. There was a linkagedisequilibrium between H297Q and -493G/T (D’ = 0.77). As the χ2 test was used, the genotype distribution of MTP -493G/T demonstrated a significant difference between the self-limited infection group and the entire chronic group or the chronic patients with no family history (χ2 = 8.543, P = 0.015 and χ2 = 7.199, P = 0.019). The allele distribution at the MTP-493 position also demonstrated a significant difference between the study groups without family history (χ2 = 6.212, P = 0.013). The T allele emerged as a possible protective factor which may influence the outcomes of HBV infection (OR: 0.59; 95% CI: 0.389-0.897). CONCLUSION: The polymorphism of the MTP gene, T allele at -493, may be involved in determining the HBV infection outcomes, of which the mechanism needs to be further investigated.

Immunogenicity and protective efficacy of DHBV DNA vaccines expressing envelope and capsid fusion proteins in ducks delivered by attenuated Salmonella typhimurium

Duck hepatitis B virus(DHBV) shares many basic characteristics with hepatitis B virus(HBV) and is an attractive model for vaccine development. In this study, DHBV DNA vaccines were designed to express envelope and capsid fusion proteins to enhance the breadth of immune response in ducks. Attenuated Salmonella typhimurium(SL7207) was used as a carrier and adjuvant to boost the magnitude of immune response. Based on this strategy, novel DNA vaccines(SL7207-p VAX1-LC and SL7207-p VAX1-SC) were generated. Growth kinetics, genetic stabilities and relative transcription levels of the L, S and C genes introduced by these vaccine strains were measured before inoculation to guarantee safety and efficacy. The relative transcript levels of the CD4 and CD8 T genes and the antibody levels(Ig Y) in ducks receiving the vaccines were higher than those in single gene delivered groups. Additionally, the copy number of covalently closed circular DNA in hepatocytes after DHBV challenge also provided evidence that our fusion vaccines could enhance the protective efficiency against DHBV infection in ducks.

RETRACTED: Prevalence and Profile of Hepatitis B Virus Infection among HIV-Infected Adults at Panzi Referral Hospital, in the Post-Conflict South Kivu Province, Eastern Democratic Republic of Congo

Short Retraction Notice? The paper does not meet the standards of "World Journal of AIDS". This article has been retracted to straighten the academic record. In making this decision the Editorial Board follows COPE's Retraction Guidelines. The aim is to promote the circulation of scientific research by offering an ideal research publication platform with due consideration of internationally accepted standards on publication ethics. The Editorial Board would like to extend its sincere apologies for any inconvenience this retraction may have caused.? Editor guiding this retraction: Prof. Linda D. Moneyham (EiC of WJA) Please see the article page for more details.? The?full retraction notice in PDF is preceding the original paper which is marked "RETRACTED".

HBV慢性感染诱导的染色体超倍化及靶向治疗策略

慢性乙型肝炎病毒感染(chronic hepatitis B infection,CHB)诱导肝细胞的染色体超倍化(包括非整倍化和多倍化)及染色体不稳定性,是导致原发性肝细胞癌(primary hepatocytic carcinoma,HCC)发生的主要原因之一.尽管肝细胞对于正常条件下染色体复制的多倍体化具有调节作用,但对于CHB引起的超倍化难以调节从而致癌.研究表明,乙型肝炎病毒(hepatitis B virus,HBV)使得多条信号途径如PLK1/PRC1失调,诱导肝细胞染色体超倍化并发生恶性转化.本论文综述了HBV感染诱导肝细胞染色体超倍化导致肝癌发生的机制以及靶向染色体超倍化药物研究的最新进展.

Baseline HBV Load Increases the Risk of Anti-tuberculous Drug-induced Hepatitis Flares in Patients with Tuberculosis

Hepatitis associated anti-tuberculous treatment（HATT） has been a main obstacle in managing patients co-infected with Mycobacterium tuberculosis and hepatitis B virus（HBV）. Therefore, we evaluated the factors related to the severity of adverse effects during HATT, especially those associated with liver failure. A retrospective study was carried out at Tongji Hospital from 2007 to 2012. Increases in serum transaminase levels of 〉3, 5, and 10 times the upper limit of normal（ULN） were used to define liver damage as mild, moderate, and severe, respectively. Patients with elevated total bilirubin（TBil） levels that were more than 10 times the ULN（〉171 μmol/L） with or without decreased（〈40%） prothrombin activity（PTA） were diagnosed with liver failure. A cohort of 87 patients was analyzed. The incidence of liver damage and liver failure was 59.8%（n=52） and 25.3%（n=22）, respectively. The following variables were correlated with the severity of hepatotoxicity： albumin（ALB） levels, PTA, platelet counts（PLT）, and the use of antiretroviral therapies（P〈0.05）. Hypo-proteinemia and antiretroviral therapy were significantly associated with liver failure, and high viral loads were a significant risk factor with an odds ratio（OR） of 2.066. Judicious follow-up of clinical conditions, liver function tests, and coagulation function, especially in patients with high HBV loads and hypoalbuminemia is recommended. It may be advisable to reconsider the use of antiviral drugs failure during the course of anti-tuberculous treatment of HBV infection patients to avoid the occurrence of furious liver failure.

Maternal Blood Milk Saliva Sample Selection and the Transmission of Hepatitis B Virus Infectious Research

Background: Hepatitis B is an infectious disease, which is a main way of vertical transmission of infectious HBV between mother and infant. Hepatitis B virus infection is always a hot topic of social concern, especially in China. The paper studies hepatitis B virus in maternal blood, breast milk, saliva of hepatitis B virus infection model (HBV-M) in Hefei city, Anhui province, PRC. HBV-DNA load and related data in Hefei city are used for risk assessment of the transmission of hepatitis B virus to provide evidence for evidence-based medicine and scientific guidance of infant feeding patterns. Methods: On the principle of informed consent, inpatient hepatitis B maternal blood 695, breast milk, saliva 614,169 copies were used as the object of analysis, using the ELISA method for the detection of HBV-M, using real-time fluorescence quantitative PCR detection of HBV-DNA load. We analyze HBsAg in saliva, milk, the positive rate of HBV-DNA and HBV-M in serum, saliva, milk, and explore the positive rate of HBV-DNA and serum HBV-DNA load correlation. Results: At the age of 18 - 44 years old perinatal women, HBV-DNA positive rates of maternal serum, breast milk, saliva were 157 cases in A group HBsAg, HBeAg positive: 99.36%, 88.06%, 96.77%;in 312 cases in group B, HBeAb HBsAg, HBcAb positive: 17.63%, 2.93%, 54.67%;69 cases in C group HBsAg, HBcAb positive: 63.77%, 27.27%, 28.57%;D group of 71 patients with simple HBcAb positive: 12.68%, 3.13%, 0%;E group and 86 cases in control group HBVM: 1.16%, 0%, 0%. According to the serum and milk testing of Group A and Group B, HBV-DNA chi-square is χ2 = 237.45, P;there is a significant difference in serum and saliva;HBV-DNA chi-square χ2 = 289.49, P < 0.01, the difference has statistical significance. Conclusion: 1) HBV-DNA load high maternal blood, breast milk, saliva are potentially persistent hepatitis B virus infection risk, especially infectious blood. 2) Of maternal milk, saliva and blood HBV-DNA HBV-DNA load were positively correlated (r = 0.96;P ing, breast milk and saliva HBV-DNA positive rates were increased and infectivity enhanced. 3) Maternal blood, breast milk, saliva specimens for any HBV-DNA ≥ 1000 copies/ml are not breastfeeding. 4) The mother who carries the hepatitis B virus cannot do maternal infant feeding, and deep kiss intimate contact, in order to prevent blood, saliva and other ways of infection of hepatitis B virus. 5) Saliva testing is instead of milk inspection, because saliva is easier;

氧化苦参碱在鸭原代肝细胞中抗鸭乙肝病毒(DHBV)作用的研究

通过建立鸭原代肝细胞-DHBV感染模型研究氧化苦参碱抗DHBV的作用。分别在DHBV感染前、感染同时以及感染后给药，利用打点杂交、Southern印迹核酸杂交和荧光定量PCR方法分别检测培养细胞上清及细胞内病毒核酸，观察氧化苦参碱在病毒感染的各个环节所起的抗病毒作用。实验结果显示：1mg／mL氧化苦参碱处理细胞后，鸭原代肝细胞培养上清及细胞内的DHBV核酸明显低于病毒感染对照组，病毒抑制率达91．6％；在病毒感染同时加药对病毒的抑制率可达98．5％；感染后持续用药能使不同培养天数的鸭肝细胞内的DHBV核酸降低60．5％～96．6％；氧化苦参碱与DHBV共孵育后，可以使病毒感染力下降69．6％。结果说明氧化苦参碱可以在DHBV感染鸭原代肝细胞的多个环节，包括病毒吸附、进入细胞及细胞内复制等方面发挥抗病毒作用。

低浓度HBsAg人群HBV DNA与HBV M定量结果的关系及评价

目的:比较高、低浓度HBsAg阳性人群乙型肝炎病毒核酸(HBV DNA)与乙型病毒肝炎标志物(HBVM)含量之间的差异,揭示低浓度HBsAg人群HBV DNA与HBV M之间的特点。方法:对264例慢性HBV感染者的HBsAg浓度(高浓度HBsAg组、低浓度HBsAg组)及自然史(免疫耐受期、免疫清除期、非活动期)进行分组,采用实时荧光聚合酶链反应(PCR)和微粒子酶免疫试验(MEIA)对147例低浓度HBsA组和117例高浓度HBsAg组的血清标本进行HBV DNA和HBV M的定量测定并进行比较。结果:低浓度HBsAg组免疫耐受期7例、免疫清除期4例血清标本HBV DNA、抗-HBs、HBeAg、抗-HBe之间的差异存在统计学意义(t=2.531～9.181,P<0.01～0.05),非活动期136例血清标本中有94.1%(128/136)HBV DNA处于105copies/L以下,直接法PCR、浓缩法PCR对HBV DNA检出率分别为10.3%(14/136)、10.3%(47/136),且HBV DNA与HBV M各指标之间均不存在相关性(P>0.05);高浓度HBsAg组免疫耐受期25例HBV DNA与HBeAg、抗HBe存在相关性(r=0.744～0.772,tr=3.858～4.207,P<0.01),免疫清除期46例和非活动期46例HBV DNA则仅与抗HBs存在负相关性(r=-0.693～-0.598,tr=-4.616～-3.936,P<0.01～0.05)。结论:低浓度HBsAg人群具有特殊的血清学特点,可能与机体免疫耐受及免疫系统个体化反应有关。

HBsAg阳性母亲与婴幼儿HBV血清标志物关系

目的了解HBsAg阳性母亲及其经乙肝高效价免疫球蛋白联合乙肝疫苗免疫的婴幼儿乙型肝炎病毒（HBV）血清标志物变化及转归情况。方法对125例HBsAg阳性母亲及其128例婴幼儿（双胎3例）进行随访调查,并分别于婴幼儿7月龄、24~36月龄应用酶联免疫吸附试验检测母亲和婴幼儿血清HBV主要标志物。结果7月龄随访83例婴幼儿,24~36月龄随访75对母子。128例婴幼儿出生时HBsAg阳性4例,随访时仅1例持续阳性,其余出生时HBsAg阴性的94例婴幼儿随访中未发现HBsAg阳转;HBsAg、HBeAg双阳性的26例母亲所产婴幼儿出生时20例HBeAg阳性,随访时除1例HBsAg为阳性的婴幼儿HBeAg仍持续阳性外,余均转阴,未见婴幼儿出现HBeAg阳转;婴幼儿抗-HBe和抗-HBc在7月龄和远期随访中逐渐阴转。结论乙肝病毒经胎盘所致的宫内感染率约为3.13%（4/128）,出生后转为慢性感染者约为25.00%（1/4）;HBeAg可通过人类胎盘从母亲传递给胎儿,但在7月龄前消失;出生于HBsAg阳性母亲的婴幼儿在婴幼儿期单独抗-HBe和（或）抗-HBc阳性,不能说明处于HBV感染状态。

原代培养人胎肝细胞体外感染HBV的研究

目的 建立HBV感染人胎肝细胞体外培养系统。 方法 首先分离、培养人胎肝细胞,然后应用HBV阳性血清感染体外培养的人胎肝细胞;每隔2d收集上清液和肝细胞,应用ELISA、免疫组化法、原位杂交法和斑点杂交法检测上清液和细胞中HBsAg和HBV DNA。 结果 上清液中HBsAg在感染后2d～20d均可测出,以感染后4d～16d达高峰(A值在0.22左右)。免疫组化检测细胞中HBsAg呈阳性表达,原位杂交和斑点杂交检测细胞和上清液中HBV DNA也呈阳性表达。 结论 HBV在原代培养人胎肝细胞中能稳定复制和表达至少达16d。