Positive Rate of Different Hepatitis B Virus Serological Markers in Peking Union Medical College Hospital,a General Tertiary Hospital in Beijing

Objectives To investigate the positive rate of different hepatitis B virus （HBV） serological markers, and the demographic factors related to HBV infection. Methods We enrolled all patients tested for HBV serological markers, such as HBV surface antigen （HBsAg）, HBV surface antibody （HBsAb）, hepatitis B e antigen （HBeAg）, hepatitis B e antibody （HBeAb）, HBV core antibody （HBcAb）, and HBV-DNA from July 2008 to July 2009 in Peking Union Medical College Hospital. The positive rate of each HBV serological marker was calculated according to gender, age, and department, respectively. The positive rates of HBV-DNA among patients with positive HBsAg were also analyzed. Results Among 27 409 samples included, 2681 （9.8%） were HBsAg positive. When patients were divided into 9 age groups, the age-specific positive rate of HBsAg was 1.2%, 9.6%, 12.3%, 10.9%, 10.3%, 9.7%, 8.0%, 5.8%, and 4.3%, respectively. The positive rate of HBsAg in non-surgical department, surgical department, and health examination center was 16.2%, 5.8%, and 4.7%, respectively. The positive rate of HBsAg of males （13.3%） was higher than that of females （7.3%, P=0.000）. Among the 2681 HBsAg （＋） patients, 1230 （45.9%） had HBV-DNA test, of whom 564 （45.9%） were positive. Patients with HBsAg （＋）, HBeAg （＋）, and HBcAg （＋） result usually had high positive rate of HBV-DNA results （71.8%, P=0.000）. Conclusions Among this group of patients in our hospital, the positive rate of HBsAg was relatively high. Age group of 20-29, males, and patients in non-surgical departments were factors associated with high positive rate of HBsAg.

Prevalence of hepatitis B virus infection and associated risk factors among adult females infected with Human Immunodeficiency Virus in Ogun State,Nigeria

Background:Hepatitis B virus(HBV)infection is prevalent in sub-Saharan Africa,including Nigeria,and is frequently observed in individuals co-infected with human immunodeficiency virus(HIV).Objective:This study aims to evaluate the prevalence of serological markers for hepatitis B virus and identify the associated risk factors among women with HIV undergoing highly active antiretroviral therapy(HAART)in Ogun State,Nigeria.Methods:Ethical approval was obtained from the Babcock University Health Research Ethics Committee(BUHREC)to recruit a total of 110 adult women infected with HIV,receiving treatment at the HIV clinics of Babcock University Teaching Hospital(BUTH)in Ilishan-Remo and General Hospital in Ijebu-Ode,both located in Ogun State,Nigeria.The participants’HIV status were confirmed using three rapid diagnostic kits:Determine(Abbott Laboratories,Tokyo,Japan),Unigold HIV(Trinity Biotech Plc Bray,Co.Wicklow,Ireland),and 1/2 Stat Pak(Abbott Laboratories,Tokyo,Japan)(Chembio Diagnostic Systems,New York,USA).Additionally,an HBV 5 in 1 Panel manufactured by Innovation Biotechnology Co.,Ltd in Beijing,China,was employed to detect HBV markers qualitatively in serum samples.Results:Out of the 110 subjects that voluntarily participated in the study,4(3.6%)tested positive for HBsAg,2(1.8%)tested positive for HBsAb,81(73.6%)tested positive for HBeAg,3(2.7%)tested positive for HBeAb,and 65(59.1%)tested positive for HBcAb.There was no significant correlation between the occurrence of HBsAg and the socio-demographic characteristics of the participants(P>0.05).Various risk factors were identified,including lack of knowledge about HBV,absence of HBV vaccination history,history of blood transfusion,organ transplant,and engaging in unprotected sex,among others.Conclusion:The findings highlight the presence of HBV infection among HIV-positive women undergoing HAART in Ogun State,Nigeria,particularly within the age groups of 18–25 years and 26–30 years.These results emphasize the necessity for continuous and targeted public health interventions among this specific population.

Epidemiological and Serological Profile of Hepatitis B Virus in an Urban Area in Mali

The objective of our study was to evaluate hepatitis B virus (HBV) infection in an urban population. This longitudinal study was conducted in Bamako District and Kati Commune. After a preparatory phase, the persons who accepted the protocol were assessed for HBsAg. HBsAg carriers had blood collection for HBeAg assay, viral load assessment, genotyping, DNA mutation testing, and severity of hepatic fibrosis and necrosis. At the end of this study, 1475 persons were included, of which 195 had HBsAg positive confirmed, that is to say 13.97%. The mean age of HBsAg positive patients was 35.11 ± 11.12 years with a sex ratio of 2.68. HBeAg was found in 8.9% of the patients tested for this antigen. The viral load was undetectable in 10.52% of patients and greater than 2000 IU/mL in 32.24% of cases. Fibrosis ≥ F2 and necrosis ≥ A2 were found in respectively 19.72% and 6.80% of cases. Genotype E was found in 91.6 patients and an R249S mutation observed in 39.04% of cases. Conclusion: HBV infection has a serious impact on socio-economic development in Mali because it affects mainly the young male population, hence the need to organize preventive measures effectively.

Elevated soluble 4-1BB is associated with serum markers of hepatitis B virus in patients with chronic hepatitis B

BACKGROUND Previous studies have suggested that the costimulatory molecule 4-1BB plays pivotal roles in regulating immunity during chronic viral infection.However,up to now,there are few studies about 4-1BB in chronic hepatitis B(CHB).AIM To clarify this issue,we report our comprehensive study results on the expression levels of 4-1BB in patients with CHB.METHODS From September 2018 to June 2019,a total of 64 patients with CHB were recruited from the Department of Hepatology,The First Hospital of Jilin University.Peripheral blood samples were collected from 52 treatment-naïve and 12 entecavir-treated patients with CHB as well as 37 healthy donors(including 24 healthy adults and 13 healthy children).The levels of soluble 4-1BB(s4-1BB)in plasma were measured by ELISA.4-1BB mRNA expression in peripheral blood mononuclear cells was detected by real-time quantitative PCR.RESULTS The s4-1BB levels in the plasma of patients with CHB were significantly higher than those in healthy adults(94.390±7.393 ng/mL vs 8.875±0.914 ng/mL,P<0.001).In addition,the s4-1BB level in plasma was significantly increased in patients with a higher viral load and a disease flare up.However,there were no significant differences between treatment-naïve and entecavir-treated patients.Interestingly,among treatment-naïve patients with CHB,the levels of s4-1BB in plasma had a significant positive correlation with hepatitis B surface antigen,hepatitis B virus DNA,hepatitis B e antigen,and triglyceride levels(r=0.748,P<0.001;r=0.406,P=0.004;r=0.356,P=0.019 and r=-0.469,P=0.007,respectively).The 4-1BB mRNA expression was higher in the peripheral blood mononuclear cells of patients with CHB than in the peripheral blood mononuclear cells of healthy adults,but the difference was not statistically significant.CONCLUSION These results suggest that the levels of s4-1BB may be associated with pathogenesis of hepatitis B virus and therefore may be a promising biomarker for disease progression.

Comparative Assay of Hepatitis B and C Virus Infection Markers by Different Assay Kits^1

In order to compare sensitivity of EIA and RIA assay kits for hepatitis B and C virus (HBV and HCV, respectively) infection markers, 100 serum samples in total were collected form 50 adult women each in urban and rural areas in northeast China. The number of positive cases to the three infection markers on HBV (i.e., HBsAg +, anti HBs +, and anti HBc +) and the one on HCV (anti HCV +) were examined in two laboratories, i.e., in Laboratory A with EIA kits produced in China and in Laboratory B with RIA kits. HCV infection positivity (anti HCV +) was examined by EIA kits in both laboratories, but from different sources in and outside of China, respectively. The assay in Laboratory A gave 2 HBsAg + cases out of the 100 cases examined, whereas there were 9 positive cases in Laboratory B. In contrast, 19 cases were positive to anti HCV when examined in Laboratory A, and there were 3 cases in Laboratory B. Thus, the kits used in Laboratory A gave fewer HBsAg + and more anti HCV + cases than the kits used in Laboratory B. The prevalence of anti HBs + or anti HBc + and cases did not differ when assayed in the two laboratories with EIA and RIA kits, respectively. The agreement of positive and negative findings between the two sets of testing were 93%, 93%, 93%, 86% and 82% for HBsAg, anti HBs, anti HBc, HBV (i.e., either positive to anyone of the three markers or negative to all three markers), and anti HCV, respectively. The implication of the observation on epidemiology on HBV and HCV infection prevalence was discussed.

不同核酸提取方法对HBV-DNA检测性能验证情况分析

目的评估2种乙型肝炎病毒(HBV)-DNA提取方法及2家检测试剂的性能,有助于选择优化提取试剂和检测试剂。方法2023年4月采用达安全自动核酸提取仪提取法(磁珠法)和手工提取法(一步法),并用达安和圣湘2种HBV-DNA试剂检测,对其进行精密度、正确度、线性范围、检出限及抗干扰能力等性能进行验证和评价。结果达安全自动核酸提取仪提取达安试剂检测、手工提取达安试剂检测和手工提取圣湘试剂检测在精密度、正确度、线性范围、检出限方面验证结果均达标;达安全自动核酸提取仪提取圣湘试剂检测在低值检测中变异系数大于5%,最低检测限验证不合格;抗干扰能力方面,全自动核酸提取仪提取的2.0 g/dL血红蛋白浓度的样本用达安和圣湘试剂检测结果均不受影响。手工提取甘油三酯浓度达3000 mg/dL的样本用达安和圣湘试剂检测的结果均不受影响。结论不同厂家的提取和检测试剂避免混用,达安和圣湘试剂对HBV-DNA定量检测的结果均符合要求。

92031例癌症患者HBV血清标志物特征分析

目的了解癌症患者乙型肝炎病毒(hepatitis B virus,HBV)的感染状态和感染特点。方法回顾性分析2017年7月26日至2023年9月18日于广西医科大学附属肿瘤医院确诊的92031例癌症患者的HBV血清标志物检测结果,以肝癌、非肝癌进行分组,比较未感染(全阴或Anti-HBs阳性)、感染(除外Anti-HBs任何一项阳性)、隐性感染(occult hepatitis B virus infection,OBI;HBsAg阴性、血清或肝组织HBV DNA阳性)的占比。结果92031例癌症患者的HBV总感染率为73.75%(67876/92031),其中肝癌患者的HBV总感染率为97.65%(8922/9137),非肝癌患者的HBV总感染率为71.12%(58954/82894),肝癌组的普通感染率和OBI率均显著高于非肝癌组(均P<0.001)。肝癌组HBV血清标志物中HBsAg、HBeAg、Anti-HBe、Anti-HBc的阳性率明显高于非肝癌组(均P<0.001),但Anti-HBs的阳性率低于非肝癌组(P<0.001)。肝癌组和非肝癌组分别有20种和27种血清标志物组合模式,其中14种模式构成比在两组间差异有统计学意义(均P<0.001);两组均有7种OBI血清组合模式,其中5种模式构成比在两组间的差异有统计学意义(均P<0.05)。结论癌症患者HBV感染状态和血清学组合模式复杂,区分肝癌与非肝癌进行HBV感染统计更利于癌症患者的HBV感染评估。

Relationship between Maternal PBMC HBV cccDNA and HBV Serological Markers and its Effect on HBV Intrauterine Transmission

Objective To investigate the relationship between maternal peripheral blood mononuclear cells (PBMC) hepatitis B virus(HBV) covalenty closed circular deoxyribonucleic acid(cccDNA) and other HBV serological markers and its effects on HBV intrauterine transmission. Methods We enrolled 290 newborns and their hepatitis B surface antigen(HBsAg) positive mothers. HBV cccDNA in PBMC and HBV DNA in serum were detected by a real‐time PCR‐TaqM an probe while HBV serological markers were detected with an electrochemiluminescence immunoassay. Results There was a positive correlation between the levels of PBMC HBV cccD NA and serum HBV DNA and HBeA g(r = 0.436 and 0.403, P < 0.001). The detection rate of pattern A [‘HBsA g(+), HBeA g(+), and anti‐HBc(+)’] was significantly higher in the PBMC HBV cccD NA positive group than in the control group(χ^2 = 48.48, P < 0.001). There was a significant association between HBV intrauterine transmission and PBMC HBV cccD NA(χ^2 = 9.28, P = 0.002). In the presence of serum HBV DNA, HBeA g, and PBMC HBV cccD NA, the risk of HBV intrauterine transmission was three times higher(OR = 3.69, 95% CI: 1.30‐10.42) than that observed in their absence. The risk of HBV intrauterine transmission was the greatest(OR = 5.89, 95% CI: 2.35‐14.72) when both PBMC HBV cccD NA and pattern A were present. A Bayesian network model showed that maternal PBMC HBV cccD NA was directly related to HBV intrauterine transmission. Conclusion PBMC HBV cccDNA may be a direct risk factor for HBV intrauterine transmission. Our study suggests that serological markers could be combined with PBMC‐related markers in prenatal testing.

Update on hepatitis B and C virus diagnosis

Viral hepatitis B and C virus(HBV and HCV) are responsible for the most of chronic liver disease worldwide and are transmitted by parenteral route, sexual and vertical transmission. One important measure to reduce the burden of these infections is the diagnosis of acute and chronic cases of HBV and HCV. In order to provide an effective diagnosis and monitoring of antiviral treatment, it is important to choose sensitive, rapid, inexpensive, and robust analytical methods. Primary diagnosis of HBV and HCV infection is made by using serological tests for detecting antigens and antibodies against these viruses. In order to confirm primary diagnosis, to quantify viral load, to determine genotypes and resistance mutants for antiviral treatment, qualitative and quantitative molecular tests are used. In this manuscript, we review the current serological and molecular methods for the diagnosis of hepatitis B and C.

Hepatitis B virus genetic mutations and evolution in liver diseases

Hepatitis B virus(HBV)belongs to the genus Orthohepadnavirus of the Hepadnaviridae family and is approximately 3.2 kb in length.Owing to a lack of proofreading capacity during reverse transcription and a high replication rate,HBV exhibits as quasispecies.To detect the genetic mutations of HBV,many methods with different sensitivities and throughputs were developed.According to documentary records,HBV mutation and evolution were important vial parameters in predicting disease progression and therapeutic outcome.In this review,we separately discussed the correlation between HBV genomic mutations in four open reading frames and liver disease progression.Since some of the results were controversial from different laboratories,it remains to be seen whether functional analyses will confirm their role in modifying the course of infection.

A comparative study on serologic profiles of virus hepatitis B

INTRODUCTIONHepatitis B viral infection, one of the most-prevalent liver disorders in China and Korea, is aserious infectious disease as it has the potential ofprogressing into liver cirrhosis and primary hepaticcarcinoma. China and Korea both belong to high-risk endemic regions of viral hepatitis[1]. TheHBsAg positive rates in China ranged from 6.9% -17.9% by age, race and test methods[2-5].

Matrix-derived serum markers in monitoring liver fibrosis in children with chronic hepatitis B treated with interferon alpha

To evaluate prospectively 4 selected serum fibrosis markers （tenascin, hyaluronan, collagen Ⅵ, TIMP-1） before, during and 12 mo after IFN treatment of children with chronic hepatitis B. METHODS： Forty-seven consecutive patients with chronic hepatitis B （range 4-16 years, mean 8 years） underwent IFN treatment （3 MU tiw for 20 wk）. Fibrosis stage and inflammation grade were assessed in a blinded fashion before and 12 mo after end of treatment. Serum fibrosis markers were determined using automated assays.RESULTS： IFN treatment improved histological inflammation but did not change fibrosis in the whole group or in subgroups. Only hyaluronan correlated significantly with histological fibrosis（r = 0.3383, P = 0.022）. Basal fibrosis markers did not differ between responders （42.5%） and nonresponders（57.5%）. During IFN treatment only serum tenascin decreased significantly in the whole group and in nonresponders. When pretreatment values were compared to values 12 mo after therapy, TIMP-1 increased in all patients and in nonresponders, and hyaluronan decreased in all patients and in responders.CONCLUSION： Tenascin reflects hepatic fibrogenesis and inflammation which decreases during IFN treatment of children with chronic hepatitis B. TIMP-1 correlates with nonresponse and hyaluronan with histological fibrosis.

Safety and efficacy of oral HD-03/ES given for six months in patients with chronic hepatitis B virus infection

AIM： To investigate the safety and efficacy of the formulation HD-03/ES capsules in the management of patients with chronic hepatitis B infection.METHODS： A total of 25 patients were recruited to the study and were given HD-03/ES, two capsules twice daily for six months. Clinical assessment of symptoms and signs were done using the ＂clinical observation table＂ once a month before and after the treatment. Biochemical investigations of total bilirubin, ALT, AST, serum protein for liver function tests were done every month after initiating treatment. Serum was analyzed for HBV markers for HBsAg, HBeAg and HBV DNA at baseline, 4 and 6 mo alter therapy using ELISA kits from Roche.RESULTS： After 6 mo of therapy with HD-03/ES, a significant reduction of ALT values from 66.5 ± 11.1 to 39.1 ± 5.2 （P 〈 0.01） and a significant HBsAg loss （52%, P 〈 0.001）, HBeAg loss （60%, P 〈 0.05） and HBV DNA loss （60%, P 〈 0.05） was observed. Adverse effects were mild and never warranted withdrawal of the drug.CONCLUSION： The results of this pilot study indicate that HD-03/ES might be a safe and effective treatment for chronic hepatitis B infection and a long-term multicentric comparator trial is warranted and under way.

抗病毒联合抗结核治疗HBV-DNA阳性初治肺结核的疗效及肝功能、IFN-γ、SSS评分相关性分析

目的 分析抗病毒联合抗结核治疗乙肝病毒的脱氧核糖核酸(HBV-DNA)阳性初治肺结核的疗效并分析患者干扰素-γ(IFN-γ)、半定量积分系统(SSS)评分与肝功能的相关性。方法 选择2022年3月—2023年3月南通大学附属如皋医院收治的120例HBV-DNA阳性初治肺结核患者作为研究对象,按随机数字表法分为2组,各60例。对照组采取常规抗结核治疗,观察组在对照组基础上联合抗病毒治疗。患者均治疗6个月,比较2组临床疗效(病灶吸收、痰菌转阴、HBV-DNA转阴情况);治疗前、治疗后肝功能、炎症因子水平;比较2组不同时点(治疗前、治疗1、3、6个月)IFN-γ、HBV-DNA、SSS评分水平;采用双变量相关性Pearson分析肝功能、IFN-γ、SSS评分的相关性。结果 观察组病灶吸收、痰菌转阴、HBV-DNA转阴患者占比高于对照组(P<0.05);治疗后,2组谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)水平高于治疗前,白细胞介素-2(IL-2)、IL-6、IL-8、肿瘤坏死因子-α(TNF-α)水平低于治疗前,观察组低于对照组(P<0.05);治疗1、3、6个月时IFN-γ水平高于治疗前,观察组低于对照组,2组时点、组间比较差异具有统计学意义(P<0.05);治疗1、3、6个月时观察组HBV-DNA水平相较于治疗前逐步降低,而对照组HBV-DNA水平相较于治疗前升高,观察组低于对照组,2组时点、组间比较差异具有统计学意义(P<0.05);治疗1、3、6个月时SSS评分高于治疗前,但观察组低于对照组,2组时点、组间比较差异具有统计学意义(P<0.05);双变量相关性Pearson分析结果显示,IFN-γ水平与HBV-DNA阳性初治肺结核患者ALT、AST、TBiL、SSS评分呈正相关(r>0,P<0.05);2组不良反应发生率比较差异无统计学意义(P>0.05)。结论 HBV-DNA阳性初治肺结核经抗病毒联合抗结核治疗能够有效提升临床治疗效果,降低机体炎症及HBV-DNA水平,避免肝脏损伤,稳定肝功能,且不会增加患者发生不良反应的风险。

慢性乙型肝炎患者HBV-DNA水平与其肝功能及乙肝标志物的关系研究

目的探讨慢性乙型肝炎患者乙型肝炎病毒脱氧核糖核酸(HBV-DNA)水平与其肝功能及乙型肝炎标志物的关系。方法选取95例慢性乙型肝炎患者为观察组,同期选择95例健康人群为对照组。均进行HBV-DNA定量检测、肝功能检测、乙型肝炎标志物[乙型肝炎病毒表面抗原(HBsAg)、乙型肝炎病毒E抗原(HBeAg)、乙型肝炎病毒e抗体(HBeAb)、乙型肝炎病毒核心抗体(HBcAb)]检测。比较观察组不同乙型肝炎标志物表达患者的HBV-DNA表达水平,观察组患者HBV-DNA和HBeAg阳性率;比较两组肝功能指标。结果在95例慢性乙型肝炎患者中HBV-DNA阳性45例(47.37%)。观察组患者根据乙型肝炎标志物阳性模式划分为大三阳模式,即HBsAg(+)、HBeAg(+)、HBcAb(+),共20例,HBV-DNA阳性16例(80.00%),定量水平2.24×10^(8) IU/ml;小三阳模式,即HBsAg(+)、HBeAb(+)/HBcAb(+),共62例,HBV-DNA阳性21例(33.87%),定量水平4.18×10^(6) IU/ml;HBsAg(+)、HBeAg(+)、HBeAb(+)、HBcAb(+)模式,共10例,HBV-DNA阳性7例(70.00%),定量水平1.83×10^(7) IU/ml;HBsAg(+)、HBcAb(+)模式,共3例,HBV-DNA阳性1例(33.33%),定量水平9.48×10^(5)IU/ml。观察组不同乙型肝炎标志物表达患者的HBV-DNA阳性率比较差异具有统计学意义(P<0.05),其中大三阳和HBsAg(+)、HBeAg(+)、HBeAb(+)、HBcAb(+)模式患者的HBV-DNA阳性率、定量水平高于其他类型。在95例观察组患者中HBV-DNA阳性45例(47.37%),阴性50例(52.63%);HBeAg(+)30例(31.58%),HBeAg(-)65例(68.42%),观察组患者HBV-DNA阳性率高于HBeAg阳性率,差异有统计学意义(χ^(2)=4.957,P<0.05)。观察组患者的谷丙转氨酶(ALT)、谷草转氨酶(AST)、谷氨酰转肽酶(GGT)水平均高于对照组,且观察组中随着患者的HBV-DNA定量水平提高,其AST、ALT、GGT水平持续升高,差异有统计学意义(P<0.05)。结论临床检测慢性乙型肝炎患者乙型肝炎病毒复制时检测其HBV-DNA的阳性率高于乙型肝炎标志物,且HBV-DNA定量水平与其肝功能损害相关,因而应定期开展定量检测,通过明确患者的病毒复制情况评估病情及治疗效果,辅助采取合理的治疗方案。

乙肝大小三阳状态、不同HBVDNA载量孕妇肝功能指标水平分析

目的研究探讨乙肝孕妇大三阳和小三阳状态中乙型肝炎病毒(Hepatitis B virus,HBV)DNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能水平。方法选取临清市人民医院于2020年1月—2023年1月收治的120例乙肝孕妇作为研究对象,根据乙肝两对半检测结果将其分为两组,即大三阳组(n=60)和小三阳组(n=60)。比较两组HBVDNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能指标、肝纤维化标志物含量。结果大三阳组丙氨酸转移酶水平(29.00±7.62)U/L、HBVDNA水平(5.18±1.16)×10^(4)U/mL均高于小三阳组的(26.00±2.85)U/L、(2.08±1.11)×10^(4)U/mL,差异有统计学意义(t=2.856、14.956,P均<0.05)。大三阳组的重组人几丁质酶3样蛋白1水平低于小三阳组,差异有统计学意义(P<0.05)。伴随着HBVDNA载量上升,两组患者甲胎蛋白水平随之增加,但是在同等HBADNA载量下,两组甲胎蛋白水平比较,差异无统计学意义(P>0.05)。结论开展孕前抗乙肝病毒诊疗、孕期风险系数评估,采取科学有效的干预措施,均能够有效抑制乙肝在母婴中的传播。

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.

失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与生化及凝血等指标相关性分析

目的分析失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与乙型肝炎病毒标志物、生化及凝血等指标的相关性。方法回顾性分析2018年1月至2020年12月我院收治的645例失代偿期乙型肝炎肝硬化患者的临床资料,根据HBV-DNA水平分为4组:阴性组、低病毒载量组(<2.0×10^(3)IU/mL)、中病毒载量组(2.0×10^(3)~2.0×10^(5)IU/mL)和高病毒载量组(>105IU/mL),测定所有患者乙型肝炎病毒血清学标志物(HBV-M)、肝功能相关生化指标(ALT、AST、TBIL、DBIL、ALB)、凝血相关指标(PT、INR、APTT、TT、D-D、PLT),比较4组患者各项指标的差异,并分析各项指标与HBV-DNA水平的相关性。结果在失代偿期乙型肝炎肝硬化患者中,HBsAg和HBeAg的定量数值随着HBV-DNA水平的升高而增高,HBsAg的明显升高主要体现在HBV-DNA阴性组和低病毒载量组之间(779.40 vs 5773.00IU/mL),而HBeAg的表达在中病毒载量组(0.19 COI)和高病毒载量组(7.96 COI)中才出现显著升高,低病毒载量组的HBeAg阳性表达率较低。随着病毒载量的升高,患者的ALT、AST、TBIL、DBIL、D/T、AFP指标均有所升高,而ALB的水平降低;除TT存在显著性差异外,PT、APTT、INR、Fib、D-D、PLT这6个指标差异均无统计学意义。经Pearson相关性分析检验,失代偿期乙型肝炎肝硬化患者血清HBV-DNA水平与ALT、AST、TBIL、DBIL、AFP、PT、APTT、TT、D-D、HBsAg、HBeAg水平呈正相关(r>0,P均<0.05),与ALB、Fib水平呈负相关(r<0,P均<0.05),与PLT、HBeAb水平不存在相关性。结论在失代偿期乙型肝炎肝硬化患者中,血清HBV-DNA载量与HBsAg、HBeAg表达水平密切相关,与ALT、AST、TBIL、DBIL、D/T、ALB等肝功能相关生化指标相关,与凝血相关指标存在关联,但在不同HBV-DNA载量组中无差异。HBV-DNA载量可作为HBeAg阴性患者肝脏损害程度的有效预测指标。

血清定量HBsAg和HBV DNA预测HBeAg阳性慢性HBV感染显著肝炎活动的性能比较

目的 对比评价血清HBsAg和HBV DNA预测HBeAg阳性慢性HBV感染显著肝炎活动的性能。方法 505例HBeAg阳性患者入选本研究。HBsAg、HBeAg采用Architect I2000免疫分析仪检测,HBV DNA采用LightCycler 480 qPCR仪检测;血清ALT采用Architect C16000生化分析仪检测。肝脏病理学分级和分期参照Scheuer标准。显著肝炎活动分三个层级,依次定义为“ALT≥20 IU/L或(分级>G1或分期>S1)”、“ALT≥30 IU/L或(分级>G1或分期>S1)”、“ALT≥40 IU/L或(分级>G1或分期>S1)”。结果 根据ALT与HBsAg、HBeAg和HBV DNA的LOESS回归分析,279例患者被界定为疑似HBV高复制人群。总体人群,HBsAg和HBV DNA预测三个层级显著肝炎活动的ROC曲线下面积(AUC)依次为0.737和0.532、0.737和0.548、0.686和0.545,疑似HBV高复制人群,HBsAg和HBV DNA预测三个层级显著肝炎活动的AUC依次为0.866和0.544、0.837和0.540、0.787和0.564。无论总体或疑似HBV高复制人群,HBsAg和HBV DNA预测三个层级显著肝炎活动的AUC分别均显著大于和均接近对角参考线下面积(P均<0.002和P均>0.150),HBsAg预测三个层级显著肝炎活动的AUC均显著大于HBV DNA(P<0.001)。以HBsAg≤4.699 log10 IU/mL为标准,其预测总体人群ALT≥20 IU/L、≥40 IU/L显著肝炎活动的灵敏度和特异度分别为86.50%和56.25%、89.52%和43.94%,预测疑似HBV高复制人群ALT≥20 IU/L、≥40 IU/L显著肝炎活动的灵敏度和特异度分别为75.37%和81.82%、80.51%和67.44%。结论 HBsAg对HBeAg阳性显著肝炎活动有良好预测意义,而HBV DNA没有预测价值。

微小RNA-127-3p在乙型肝炎病毒相关性肝癌中的表达意义及对HBV阳性肝癌细胞增殖迁移侵袭的影响

目的:探讨微小RNA-127-3p(miR-127-3p)在乙型肝炎病毒(HBV)相关性肝癌中的表达意义及其对HBV阳性肝癌细胞增殖、迁移和侵袭的影响。方法:采用实时荧光定量聚合酶链反应(RTFQ-PCR)检测2018年12月至2020年12月在医院手术切除的64例HBV相关性肝癌患者肝癌组织和癌旁组织中miR-127-3p表达量。将对数生长期肝癌HepG2细胞分别以10、20、50 pmol的miR-127-3p mimic进行转染,同时设置对照组(以0 pmol mimic进行转染),培养48 h后采用RTFQ-PCR检测各转染组细胞中miR-127-3p表达量,MTT法检测各转染组细胞增殖情况,细胞划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力。结果:肝癌组织miR-127-3p表达量(0.26±0.03)低于癌旁组织(0.53±0.06)(P<0.05)。miR-127-3p mimic转染组(10、20、50 pmol)与对照组肝癌HepG2细胞中miR-127-3p表达量分别为(0.87±0.09)、(1.23±0.13)、(1.51±0.16)、(0.28±0.04),总体比较差异有统计学意义(P<0.05);与对照组比较,miR-127-3p mimic(10、20、50 pmol)转染组miR-127-3p表达量更高(P<0.05);miR-127-3p mimic转染组中miR-127-3p表达量随转染浓度的升高而升高(P<0.05)。miR-127-3p mimic转染组(10、20、50 pmol)与对照组肝癌HepG2细胞增殖率分别为[(51.58±5.24)%、(25.42±2.55)%、(16.23±1.65)%、(78.67±7.88)%],细胞迁移距离分别为[(28.26±2.84)mm、(23.17±2.25)mm、(18.32±1.83)mm、(33.87±3.41)mm],穿膜细胞数分别为[(122.54±12.26)个、(101.26±10.45)个、(46.15±4.78)个、(131.23±13.24)个],总体比较差异均有统计学意义(P<0.05);与对照组比较,miR-127-3p mimic(10、20、50 pmol)转染组细胞增殖率更低,细胞迁移距离更短,穿膜细胞数更少(P<0.05);miR-127-3p mimic转染组细胞增殖率随转染浓度的升高而降低,细胞迁移距离随转染浓度的升高而变短,穿膜细胞数随转染浓度的升高而减少(P<0.05)。结论:miR-127-3p在HBV相关性肝癌患者中呈低表达,其可抑制HBV阳性肝癌HepG2细胞增殖、迁移和侵袭。