Hepatitis B virus X protein promotes liver cell proliferation via a positive cascade loop involving arachidonic acid metabolism and p-ERK1/2

Hepatitis B virus X protein （HBx） plays a crucial role in the development of hepatocellular carcinoma. Here, we sought to identify the mechanisms by which HBx mediates liver cell proliferation. We found that HBx upregulated the levels of cyclooxygenase-2 （COX-2）, 5-1ipoxygenase （5-LOX） and phosphorylated extracellular signal-regulated protein kinases 1/2 （p-ERK1/2） in liver cells. HBx-induced p-ERK1/2 was abolished by inhibition of Gi/o proteins, COX or LOX. In addition, HBx increased the amounts of prostaglandin E2 （PGE2） and leukotriene B4 （LTB4） released from cell lines derived from hepatocytes. Moreover, these released arachidonic acid metabolites were able to activate ERK1/2. Interestingly, activated ERK1/2 could upregulate the expression of COX-2 and 5-LOX in a positive feedback manner. In conclusion, HBx enhances and maintains liver cell proliferation via a positive feedback loop involving COX-2, 5-LOX, released arachidonic acid metabolites, Gi/o proteins and p-ERK1/2.

Antiviral effects of hepatitis B virus S gene-specific anti-gene locked nucleic acid in transgenic mice

AIM To assess the antiviral effects of hepatitis B virus(HBV) S gene-specific anti-gene locked nucleic acid(LNA) in transgenic mice.METHODS Thirty HBV transgenic mice were acclimatized to laboratory conditions and positive for serum HBV surface antigen(HBs Ag) and HBV DNA, were randomly divided into 5 groups(n = 7), including negative control(blank control, unrelated sequence control), positive control(lamivudine, anti-sense-LNA), and anti-gene-LNA experimental group. LNA was injected into transgenic mice by tail vein while lamivudine was administeredby gavage. Serum HBV DNA and HBs Ag levels were determined by fluorescence-based PCR and enzymelinked immune sorbent assay, respectively. HBV S gene expression amounts were assessed by reverse transcription polymerase chain reaction. Positive rates of HBsA g in liver cells were evaluated immunohistochemistry.RESULTS Average rate reductions of HBs Ag after treatment on the 3 rd, 5 th, and 7 th days were 32.34%, 45.96%, and 59.15%, respectively. The inhibitory effect of antigene-LNA on serum HBs Ag peaked on day 7, with statistically significant differences compared with pretreatment(0.96 ± 0.18 vs 2.35 ± 0.33, P < 0.05) and control values(P < 0.05 for all). Average reduction rates of HBV DNA on the 3 rd, 5 th, and 7 th days were 38.55%, 50.95%, and 62.26%, respectively. This inhibitory effect peaked on the 7 th day after treatment with anti-gene-LNA, with statistically significant differences compared with pre-treatment(4.17 ± 1.29 vs 11.05 ± 1.25, P < 0.05) and control values(P < 0.05 for all). The mR NA levels of the HBV S gene(P < 0.05 for all) and rates of HBsA g positive liver cells(P < 0.05 for all) were significantly reduced compared with the control groups. Liver and kidney function, and histology showed no abnormalities. CONCLUSION Anti-gene-LNA targeting the S gene of HBV displays strong inhibitory effects on HBV in transgenic mice, providing theoretical and experimental bases for gene therapy in HBV.

The effect of mycophenolate acid on hepatitis B virus replication in vitro

OBJECTIVE: To use 2.2.15 cell line to determine the effects of mycophenolate acid (MPA) on hepatitisB virus (HBV) replication and viral protein synthesis in vitro.METHODS: The 2.2.15 cells were treated with different concentration of MPA (1-50 μg/ml) for 12days. HBsAg and HBeAg were detected in the supernatant fluid by ELISA and intracellular HBV DNAwas analyzed quantitatively by slot blot hybridization.RESULTS: MPA could suppress the expression of HBsAg and HBeAg, and the higher concentration ofMPA induced lower expression of HBsAg and HBeAg. The suppression rates of MPA for HBsAg andHBeAg at a concentration of 50 μg/ml were 34.2% and 24.1% respectively. The expression of HBVDNA was only 49% as compared with controls when treated with MPA at a concentration of 50 μg/ml.CONCLUSIONS: Mycophenolate acid can suppress the expression of HBsAg and HBeAg as well as thereplication of HBV DNA in the 2.2.15 cell. The suppressive degree is dose-dependent.

Antiviral Effects of Stichopus japonicus Acid Mucopolysaccharide on Hepatitis B Virus Transgenic Mice

Hepatitis B virus(HBV) is a significant global pathogen and efficient cure for HBV patients is still a challenging goal. We previously reported that acidic mucopolysaccharide from stichopus japonicus selenka(SJAMP) could inhibit HBs Ag and HBe Ag expression in vitro. However, the potential anti-HBV effects of SJAMP in vivo have not yet been explored. In this study, we show that SJAMP exhibits potent anti-HBV activity in HBV transgenic mice in a dose-dependent manner. Specifically, sixty HBV transgenic male BALB/c mice were randomly selected to receive the treatment of PBS, low dose SJAMP(30 mg kg^(-1)), middle dose SJAMP(40 mg kg^(-1)), high dose SJAMP(50 mg kg^(-1)) and IFN(45 IU kg^(-1)) for 30 d. SJAMP treatment suppressed serum HBV-DNA, and liver HBs Ag and HBc Ag levels in HBV-transgenic mice. The present study highlights the potential application of SJAMP in HBV therapy.

Impact of hepatitis B virus infection on hepatic metabolic signaling pathway

A growing body of epidemiologic research has demonstrated that metabolic derangement exists in patients with hepatitis B virus(HBV) infection, indicating that there are clinical associations between HBV infection and host metabolism. In order to understand the complex interplay between HBV and hepatic metabolism in greater depth, we systematically reviewed these alterations in different metabolic signaling pathways due to HBV infection. HBV infection interfered with most aspects of hepatic metabolic responses, including glucose, lipid, nucleic acid, bile acid and vitamin metabolism. Glucose and lipid metabolism is a particular focus due to the significant promotion of gluconeogenesis, glucose aerobic oxidation, the pentose phosphate pathway, fatty acid synthesis or oxidation, phospholipid and cholesterol biosynthesis affected by HBV. These altered metabolic pathways are involved in the pathological process of not only hepatitis B, but also metabolic disorders, increasing the occurrence of complications, such as hepatocellular carcinoma and liver steatosis. Thus, a clearer understanding of the hepatic metabolic pathways affected by HBV and its pathogenesis is necessary to develop more novel therapeutic strategies targeting viral eradication.

Differentially expressed genes in hepatocellular carcinoma induced by woodchuck hepatitis B virus in mice

INTRODUCTIONHepatocellular carcinoma(HCC)is one of the major causes of death in the word.The mechanism of carcinogenesis is unknown,although it is widely accepted that HBV and HCV are clsely related to liver cancer[1-5[1-5].Previously,a variety of studies have described the differences in gene expression which distinguished tumor from nontumor[6-11].Cloning of the genes,especially the genes associated with HBV and HCV,is still very important to account for the development of liver cancer.

Effects of SAHA on proliferation and apoptosis of hepatocellular carcinoma cells and hepatitis B virus replication

AIM: To investigate the effects of suberoylanilide hydroxamic acid(SAHA) on proliferation and apoptosis of a human hepatocellular carcinoma cell line(HepG2.2.15) and hepatitis B virus(HBV) replication.METHODS: HepG2.2.15 cells were treated with different concentrations of SAHA.Cell morphology was examined by confocal laser scanning microscopy,and cell proliferation was determined using a MTT colorimetric assay.Flow cytometry was used to detect apoptosis and determine cell cycle phase,while hepatitis B surface antigen and hepatitis B e antigen content were measured using chemiluminescence.Reverse transcription polymerase chain reaction was performed to measure HBV DNA in cell lysate.RESULTS: Cell proliferation rates were significantly reduced by the addition of SAHA.The inhibitory effect of SAHA on cell proliferation was both time-and dosedependent.After 24 h of treatment with SAHA,the early cell apoptotic rate increased from 3.25% to 21.02%(P = 0.041).The proportion of G0 /G1 phase cells increased from 50.3% to 65.3%(P = 0.039),while that of S phase cells decreased from 34.9% to 20.6%(P = 0.049).After 48 h of treatment,hepatitis B surface antigen and hepatitis B e antigen content increased from 12.33 ± 0.62 to 25.42 ± 2.67(P = 0.020) and 28.92 ± 1.24 to 50.48 ± 1.85(P = 0.026),respectively.Furthermore,HBV DNA content increased from 4.54 ± 0.46 to 8.34 ± 0.59(P = 0.029).CONCLUSION: SAHA inhibits HepG2.2.15 cell proliferation,promotes apoptosis,and stimulates HBV replication.In combination with anti-HBV drugs,SAHA may potentially be used cautiously for treatment of hepatocellular carcinoma.

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;

Gene therapeutic approaches to inhibit hepatitis B virus replication

Acute and chronic hepatitis B virus(HBV) infections remain to present a major global health problem. The infection can be associated with acute symptomatic or asymptomatic hepatitis which can cause chronic inflammation of the liver and over years this can lead to cirrhosis and the development of hepatocellularcarcinomas. Currently available therapeutics for chronically infected individuals aim at reducing viral replication and to slow down or stop the progression of the disease. Therefore, novel treatment options are needed to efficiently combat and eradicate this disease. Here we provide a state of the art overview of gene therapeutic approaches to inhibit HBV replication. We discuss non-viral and viral approaches which were explored to deliver therapeutic nucleic acids aiming at reducing HBV replication. Types of delivered therapeutic nucleic acids which were studied since many years include antisense oligodeoxynucleotides and antisense RNA, ribozymes and DNAzymes, RNA interference, and external guide sequences. More recently designer nucleases gained increased attention and were exploited to destroy the HBV genome. In addition we mention other strategies to reduce HBV replication based on delivery of DNA encoding dominant negative mutants and DNA vaccination. In combination with available cell culture and animal models for HBV infection, in vitro and in vivo studies can be performed to test efficacy of gene therapeutic approaches. Recent progress but also challenges will be specified and future perspectives will be discussed. This is an exciting time to explore such approaches because recent successes of gene therapeutic strategies in the clinic to treat genetic diseases raise hope to find alternative treatment options for patients chronically infected with HBV.

2D-QSAR Studies on Anthranilic Acid Derivatives: A Novel Class of Allosteric Inhibitors of Hepatitis C NS5B Polymerase

Quantitative structure activity relationship （QSAR） studies were performed on 45 anthranilic acid derivatives for their potent allosteric inhibition activities of HCV NSSB polymerase. Genetic algorithm based genetic function approximation （GFA） method of variable selection was used to generate the model. Highly statistically significant model with r^2 = 0.966 and r^2cv = 0.951 was obtained when the number of descriptors in the equation was set to 5. High r^2pred value of 0.884 indicates the good predictive power of the best model. Spatial descriptors of radius of gyration （RadOfGration）, molecular volume （Vm）, length of molecule in the z dimension （Shadow-Zlength）, thermodynamic descriptors of the octanol/water partition coefficient （LogP） and molecular refractivity index （MR） showed enormous contributions to HCV NS5B polymerase inhibition. The validation of the model was done by leave-one-out （LOO） test, randomization tests and external test set prediction. The model gives insight on indispensable structural requirements for the activity and can be used to design more potent analogs against HCV NSSB polymerase.

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。

慢性乙型肝炎者外周血SAA/CRP、NLR水平与HBV-DNA载量、病情程度的相关性分析

目的探究慢性乙型肝炎(chronic hepatitis B,CHB)患者外周血淀粉样蛋白A(serum amyloid A,SAA)与C反应蛋白(C-reactive protein,CRP)比值(SAA/CRP)、中性粒细胞与淋巴细胞比值(neutrophils lymphocytes ratio,NLR)水平与乙型肝炎病毒-脱氧核糖核酸(HBV-DNA)载量及病情程度的相关性。方法选取2020年6月至2022年6月达州市中西医结合医院100例CHB患者作为研究组,根据病情程度分为轻度(单纯CHB,n=36)、中度(乙肝代偿期肝硬化,n=33)和重度(乙肝失代偿期肝硬化,n=31)。另选同期、同年龄段50例健康志愿者作为对照组,比较研究组不同病情程度、对照组一般资料、血清SAA/CRP、NLR水平,并比较研究组不同HBV-DNA载量患者血清SAA/CRP、NLR水平,分析CHB患者血清SAA/CRP、NLR水平与HBV-DNA载量、病情程度的相关性;所有患者均行抗病毒治疗,治疗24周,比较不同抗病毒疗效患者治疗前、治疗后12周、24周血清SAA/CRP、NLR水平及变化值,分析治疗前后血清SAA/CRP、NLR水平变化值预测疗效的价值。结果重度CHB患者血清SAA/CRP、NLR水平>中度CHB患者>轻度CHB患者>健康人群(P<0.05);高载量患者血清SAA/CRP、NLR水平>中载量患者>轻载量患者(P<0.05);CHB患者血清SAA/CRP、NLR水平与HBV-DNA载量(r=0.756、0.709)、病情程度(r=0.776、0.745)呈正相关(P<0.05);无应答患者治疗后12周、24周外周血SAA/CRP、NLR水平均高于应答患者,变化值均低于应答患者(P<0.05);SAA/CRP△1、NLR△1单独预测的AUC分别为0.752、0.773,联合预测△1的AUC为0.861;SAA/CRP△2、NLR△2单独预测的AUC分别为0.796、0.819,联合预测△2的AUC为0.967,大于联合预测△1的AUC(P<0.05)。结论CHB患者的SAA/CRP、NLR与CHB HBV-DNA载量及病情程度具有相关性,临床可通过其水平变化评估病情及预测预后。

慢性乙型肝炎患者接受核(苷)酸类似物抗病毒治疗后血清HBV-DNA的表达及临床意义

目的探讨慢性乙型肝炎(CHB)患者接受核(苷)酸类似物(NAs)抗病毒治疗后血清乙型肝炎病毒脱氧核糖核酸(HBV-DNA)的表达及临床意义。方法回顾性分析2021年1月至2022年10月河南省人民医院83例采用NAs抗病毒治疗的CHB患者临床资料,根据血清学应答标准将其分为应答组(46例)和未应答组(37例)。比较两组基线资料、治疗前及治疗3、6、12个月时血清HBV-DNA水平;绘制受试者工作特征(ROC)曲线,以曲线下面积(AUC)检验血清HBV-DNA对CHB患者NAs抗病毒治疗未应答的预测价值。结果治疗前,应答组和未应答组HBV-DNA比较,差异无统计学意义(P>0.05);两组治疗前至治疗12个月的HBV-DNA呈下降趋势,组间、时点、交互效应有统计学意义(P<0.05)。绘制ROC曲线显示,治疗3个月时HBV-DNA对CHB患者NAs抗病毒治疗未应答的预测价值较低(AUC=0.694,P=0.002),治疗6个月时HBV-DNA对CHB患者NAs抗病毒治疗未应答具有一定预测价值(AUC=0.751,P<0.001)。结论血清HBV-DNA表达在CHB患者NAs抗病毒治疗前后变化明显,且治疗6个月时血清HBV-DNA可作为抗病毒治疗未应答的预测指标。

Humoral and cellular immunogenecity of DNA vaccine based on hepatitis B core gene in rhesus monkeys

INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant association among persistentinfection, liver cirrhosis and hepatocellularcarcinoma[1-3].

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。

Conservation and variability of hepatitis B core at different chronic hepatitis stages

BACKGROUND Since it is currently not possible to eradicate hepatitis B virus(HBV)infection with existing treatments,research continues to uncover new therapeutic strategies.HBV core protein,encoded by the HBV core gene(HBC),intervenes in both structural and functional processes,and is a key protein in the HBV life cycle.For this reason,both the protein and the gene could be valuable targets for new therapeutic and diagnostic strategies.Moreover,alterations in the protein sequence could serve as potential markers of disease progression.AIM To detect,by next-generation sequencing,HBC hyper-conserved regions that could potentially be prognostic factors and targets for new therapies.METHODS Thirty-eight of 45 patients with chronic HBV initially selected were included and grouped according to liver disease stage[chronic hepatitis B infection without liver damage(CHB,n=16),liver cirrhosis(LC,n=5),and hepatocellular carcinoma(HCC,n=17)].HBV DNA was extracted from patients’plasma.A region between nucleotide(nt)1863 and 2483,which includes HBC,was amplified and analyzed by next-generation sequencing(Illumina Mi Seq platform).Sequences were genotyped by distance-based discriminant analysis.General and intergroup nt and amino acid(aa)conservation was determined by sliding window analysis.The presence of nt insertion and deletions and/or aa substitutions in the different groups was determined by aligning the sequences with genotype-specific consensus sequences.RESULTS Three nt(nt 1900-1929,2249-2284,2364-2398)and 2 aa(aa 117-120,159-167)hyper-conserved regions were shared by all the clinical groups.All groups showed a similar pattern of conservation,except for five nt regions(nt 1946-1992,2060-2095,2145-2175,2230-2250,2270-2293)and one aa region(aa 140-160),where CHB and LC,respectively,were less conserved(P<0.05).Some group-specific conserved regions were also observed at both nt(2306-2334 in CHB and 1935-1976 and 2402-2435 in LC)and aa(between aa 98-103 in CHB and 28-30 and 51-54 in LC)levels.No differences in insertion and deletions frequencies were observed.An aa substitution(P79 Q)was observed in the HCC group with a median(interquartile range)frequency of 15.82(0-78.88)vs 0(0-0)in the other groups(P<0.05 vs CHB group).CONCLUSION The differentially conserved HBC and HBV core protein regions and the P79 Q substitution could be involved in disease progression.The hyper-conserved regions detected could be targets for future therapeutic and diagnostic strategies.

TMA技术在献血者HBV-DNA检测中的应用

目的:了解无偿献血人群乙型肝炎病毒(hepatitis B virus,HBV)感染情况,评估TMA技术应用于献血者HBV-DNA筛查的效果及其必要性。方法:采用平行检测ELISA/NAT模式,对2016年3月-2018年2月169160人(次)献血者及部分归队献血者进行常规血清学和NAT检测,对NAT筛查阳性标本行核酸鉴别试验;对单边试剂HBsAg+、HBV-DNA-献血者进行中和确证试验。结果:169160人(次)献血者中,双边试剂HBsAg检测阳性的为803例,占0.476%,单边试剂检测阳性的为243例,占0.144%。对40名HBV-DNA-、单边试剂检测HBsAg+标本经中和确证试验确证,仅有4名为阳性,确证阳性率为10%。检出1003例HBV-DNA+标本,HBsAg和HBV-DNA均为阳性的739例,2者一致率为73.7%。3种试剂阳性检出率比较有统计学差异(P<0.05);Murex试剂和新创试剂2种试剂检出阳性率有统计学差异(P<0.125)。2016年3月-2017年2月和2017年3月-2018年2月期间HBsAg-/HBV-DNA+检出率比较,没有统计学差异(P>0.05)。60名HBsAg-/HBV-DNA+归队献血者中有1名发生HBsAg阳转,该名献血者为HBV窗口期感染;其余59名献血者HBsAg阴性;HBVDNA检测显示,28名献血者HBV-DNA-,31名献血者HBV-DNA+,1名结果为不确定。结论:结合HBsAg检测,常规应用TMA技术检测HBV-DNA能缩短HBV感染的检测窗口期,检出HBV隐匿性感染,避免HBV漏检,从而有效地降低输血后传播乙型肝炎潜在风险。

乙肝表面抗原双试剂阳性HBV-DNA核酸检测阴性结果分析

目的通过分析献血者乙肝表面抗原检测双试剂阳性而核酸检测阴性的结果,探讨1遍血清学联合1遍核酸检测的新检测策略对血液安全的意义。方法对2017—2019年献血者标本中乙肝表面抗原血清学双试剂阳性的标本,无论核酸检测系统采取的是混样模式还是单检模式都同时进行核酸检测。阴性的标本使用原检测系统再进行重复检测。结果 2017—2019年献血者检测标本共计25.8万人份,乙肝表面抗原双试剂阳性标本共计195份,核酸HBV DNA阳性标本172份,阴性标本23份,其中核酸单检系统阴性标本8份,核酸混检系统阴性标本15份。单检系统的8份标本再次进行检测,至少有1次为阳性的标本有5份,混检系统的15份标本再次进行检测,至少有1次为阳性的标本有9份。结论无论单检核酸检测系统还是混检核酸检测系统都会出现乙肝表面抗原双试剂阳性而核酸检测阴性的结果,其中混检系统较单检系统的标本数量要多近1倍。重复检测结果为阳性标本的比例约为60%,为低浓度标本的机会性检出。因此采取1遍血清学联合1遍核酸进行乙肝检测的新检测策略,无论是核酸检测系统还是血清学检测系统都应进行系统的评估,比较与实验室原有检测策略的差异,避免漏检的发生,减低输血传播疾病的风险。

恩替卡韦联合乳果糖治疗乙型肝炎后肝硬化的临床效果及对血清HBV-DNA、KLF2和TNFSF15的影响

目的探讨恩替卡韦联合乳果糖治疗乙型肝炎后肝硬化的临床效果及对血清乙型肝炎病毒(HBV)-DNA、锌指样转录因子2(krüppel-like factor 2,KLF2)和肿瘤坏死因子超家族成员15(tumor necrosis factor superfamily member 15,TNFSF15)的影响。方法选取2017年1月—2019年1月收治的190例乙型肝炎后肝硬化,按照治疗方法的不同,分为观察组与对照组,每组各95例。观察组予恩替卡韦+乳果糖+常规治疗,对照组予恩替卡韦+常规治疗。两组均治疗6个月。记录治疗后6个月的临床疗效,检测治疗前、治疗后6个月的肝功能相关指标[丙氨酸转氨酶(ALT)、总胆红素(TB)]、肝纤维化相关指标[透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PC-Ⅲ)]及细胞因子相关指标(KLF2、TNFSF15)的水平变化,比较治疗前及治疗后3、6个月血清HBV-DNA水平,观察不良反应发生情况。结果与对照组比较,观察组治疗总有效率升高,差异有统计学意义(P<0.05)。与对照组比较,观察组治疗后6个月ALT、TB、HA、LN、PC-Ⅲ、KLF2水平下降,TNFSF15水平升高,差异有统计学意义(P<0.01);与本组治疗前比较,两组治疗后6个月ALT、TB、HA、LN、PC-Ⅲ、KLF2水平下降,TNFSF15水平升高,差异有统计学意义(P<0.01)。与对照组比较,观察组治疗后3、6个月血清HBV-DNA水平下降,差异有统计学意义(P<0.01);与本组治疗前比较,两组治疗后3、6个月血清HBV-DNA水平下降,差异有统计学意义(P<0.01);与本组治疗后3个月比较,两组治疗后6个月血清HBV-DNA水平下降,差异有统计学意义(P<0.01)。两组不良反应总发生率比较差异无统计学意义(P>0.05),均予对症处理后症状消失。结论恩替卡韦联合乳果糖治疗乙型肝炎后肝硬化的临床效果较好,可改善肝功能,抑制肝纤维化,降低血清HBV-DNA水平,调控细胞因子的表达,且不良反应未明显增加。

一种免核酸提取HBV-DNA荧光定量PCR试剂盒临床应用价值的评估

目的评估一种全新的免核酸提取的荧光定量PCR试剂检测HBV-DNA的临床应用价值。方法通过随机检测62例临床血清样品,比较免核酸提取和核酸提取两种试剂HBV-DNA扩增结果的一致性、灵敏度和相关性。结果两种HBV-DNA荧光定量试剂阴阳结果符合率为100%;HBV-DNA定量结果无显著差异(t=1.006,P>0.05);免核酸提取法不同浓度梯度相关系数r=-0.9999(P<0.001);灵敏度为4×102IU/ml,CV<6%。结论该免核酸提取HBV-DNA荧光定量PCR试剂扩增效率、线性关系和重现性良好,具有潜在的临床应用价值和发展前景。