Sequence analysis of sub-genotype D hepatitis B surface antigens isolated from Jeddah,Saudi Arabia

Little is known about the prevalence of HBV genotypes/sub-genotypes in Jeddah province, although the hepatitis B virus (HBV) was identified as the most predominant type of hepatitis in Saudi Arabia. To characterize HBV genotypes/sub-genotypes, serum samples from 15 patients with chronic HBV were collected and subjected to HBsAg gene amplification and sequence analysis. Phylogenetic analysis of the HBsAg gene sequences revealed that 11 (48%) isolates belonged to HBV/D while 4 (18%) were associated with HBV/C. Notably, a HBV/D sub-genotype phylogenetic tree identified that eight current isolates (72%) belonged to HBV/D1, whereas three isolates (28%) appeared to be more closely related to HBV/D5, although they formed a novel cluster supported by a branch with 99% bootstrap value. Isolates belonging to D1 were grouped in one branch and seemed to be more closely related to various strains isolated from different countries. For further determination of whether the three current isolates belonged to HBV/D5 or represented a novel sub-genotype, HBV/DA, whole HBV genome sequences would be required. In the present study, we verified that HBV/D1 is the most prevalent HBV sub-genotype in Jeddah, and identified novel variant mutations suggesting that an additional sub-genotype designated HBV/DA should be proposed. Overall, the results of the present HBsAg sequence analyses provide us with insights regarding the nucleotide differences between the present HBsAg/D isolates identified in the populace of Jeddah, Saudi Arabia and those previously isolated worldwide. Additional studies with large numbers of subjects in other areas might lead to the discovery of the specific HBV strain genotypes or even additional new sub-genotypes that are circulating in Saudi Arabia.     

In Vitro Anti-hepatitis B Virus Activity of 2',3'-Dideoxyguanosine

2',3'-dideoxyguanosine(DoG) has been demonstrated to inhibit duck hepatitis B virus(DHBV) replication in vivo in a duck model of HBV infection. In the current study, the in vitro antiviral effects of DoG on human and animal hepadnaviruses were investigated. Our results showed that DoG effectively inhibited HBV, DHBV, and woodchuck hepatitis virus(WHV)replication in hepatocyte-derived cells in a dose-dependent manner, with 50% effective concentrations(EC50) of 0.3 ± 0.05, 6.82 ± 0.25, and 23.0 ± 1.5 lmol/L, respectively. Similar to other hepadnaviral DNA polymerase inhibitors,DoG did not alter the levels of intracellular viral RNA but induced the accumulation of a less-than-full-length viral RNA species, which was recently demonstrated to be generated by RNase H cleavage of pgRNA. Furthermore, using a transient transfection assay, DoG showed similar antiviral activity against HBV wild-type, 3TC-resistant rtA181 V, and adefovirresistant rtN236T mutants. Our results suggest that DoG has potential as a nucleoside analogue drug with anti-HBV activity.     

肝癌乙型肝炎病毒感染患者术后恩替卡韦抗病毒治疗的临床疗效及安全性评价

目的:探讨肝癌乙型肝炎病毒(HBV)感染患者根治性切除术后采用恩替卡韦抗病毒治疗的临床疗效及安全性。方法:收集2015年1月-2017年8月在我院行根治性切除术的肝癌HBV感染患者279例为研究对象,以血清HBV-DNA载量10~5 copies/ml为界限,分为高病毒复制组128例,低病毒复制组151例,按照随机数字表法将高病毒复制组分为高-治疗组64例、高-对照组64例,将低病毒复制组分为低-治疗组76例、低-对照组75例。高-治疗组和低-治疗组术后给予恩替卡韦0.5 mg/d,高-对照组和低-对照组未行抗病毒治疗。比较手术前、术后7 d各组的血清HBV-DNA水平,血清白蛋白(ALB)、谷丙转氨酶(ALT)、前白蛋白(PA),以及术后并发症的发生情况。结果:高-治疗组、高-对照组、低-治疗组、低-对照组术后血清ALB、PA均较治疗前降低,血清ALT均较治疗前升高,且高-治疗组或低-治疗组术后血清ALB、PA均高于高-对照组或低-对照组,血清ALT水平均低于高-对照组或低-对照组,差异均有统计学意义(P0.05);高-治疗组或低-治疗组术后血清HBV-DNA水平均低于治疗前,且均低于同期高-对照组或低-对照组,差异均有统计学意义(P0.05)。高-治疗组与高-对照组、低-治疗组与低-对照组患者术后并发症发生率均无统计学差异(P0.05)。结论:恩替卡韦能显著改善肝癌HBV感染患者术后的血清HBV-DNA载量水平和肝功能,安全性高,值得临床推广。     

乙型肝炎导向干扰素抗乙型肝炎病毒作用的体外实验研究

为提高ＩＦＮ抗ＨＢＶ疗效,实现感染细胞水平上的抗病毒性肝炎导向治疗,我们将抗ＨＢｓＡｇ单克隆抗体（ＭｃＡｂ）的Ｆ（ａｂ＇）２片段与ＩＦＮ文联,所得交联体称为乙型肝炎导向干扰素（Ｔ－ＩＦＮ）。应用ＨＢＶＤＮＡ转染细胞系（２,２,１５细胞）作模型,对Ｔ－ＩＦＮ进行了体外抗ＨＢＶ实验：通过检测细胞上清液ＨＢｓＡｇ、ＨＢｅＡｇ和ＨＢＶＤＮＡ水平的变化,结合细胞存活率来评价其抗ＨＢＶ活性,并与游离ＩＦＮ和ＭｃＡｂ以及二者的混合法（Ｍｉｘ）进行比较。结果显示,经１２天作用,Ｔ－ＩＦＮ在细胞存活率、对ＨＢｓＡｇ和ＨＢｅＡｇ抑制率及时ＨＢＶＤＮＡ抑制作用等方面均优于游离崳桑疲巍ⅲ停悖粒夂停停椋裕椋疲魏鲜逝ǘ任保担埃啊常埃埃埃桑眨恚欤谡庖慌ǘ认拢赴婊盥省荩罚梗叮ィ龋拢螅粒缫种坡剩担罚场叮罚叮ィ龋拢澹粒缫种坡剩矗埃薄担保玻ィ危模猎冢保妫缢揭韵拢欢嘤εǘ鹊模桑疲巍ⅲ停悖粒夂停停椋模危猎冢欤穑缢揭陨希龋拢螅粒绾停龋拢澹粒缫种坡示陀冢裕桑疲巍Ｌ崾荆砸唬桑疲慰赡茉诓《荆危模粮粗啤⒉《镜鞍缀铣杀泶锏榷嘞罨方谏戏⒒右种谱饔谩＃裕桑疲慰梗龋幔中Ч庞冢桑疲巍     

ina; 

ＨａＳＮＰＶ蛋白激酶基因的核苷酸序列研究张传溪＊王根＊胡萃＊吴祥甫（中国科学院上海生物化学研究所,上海２０００３１;＊浙江农业大学应用昆虫学研究所,杭州３１００２９）关键词棉铃虫;核多角体病毒;蛋白激酶基因;核苷酸序列昆虫杆状病毒是重要的生物杀虫剂,...     

Isolation of NotI clusters hypomethylated in HBV-integrated hepatocellular carcinomas by two-dimensional electrophoresis.

To examine genetic and epigenetic alterations associated with HBV integration in hepatocarcinogenesis, we compared genomic DNA profiles of primary hepatocellular carcinomas (HCCs) and cell lines that either contained or did not contain integrated HBV. To accomplish this, we carried out Restriction Landmark Genomic Scanning (RLGS), a two-dimensional system that displays 2000-3000 Not I landmark sites in a single gel electrophoresis experiment. We identified one Not I landmark spot that showed high signal intensity in HBV-integrated cell lines or in primary HCCs, but not in HCCs or tumor-cell lines free of HBV integration. Cloning of this spot revealed that it consisted of a Not I cluster sequence enriched with CpG dinucleotides. This sequence, hypomethylated in association with HBV integration, was found in the peri-centromeric region of human acrochromosomes. The results demonstrate that epigenetic changes at specific sequences in the genome occur in association with HBV integration during the process of hepatocarcinogenesis.     

乙型肝炎病毒的核心启动子各区段功能的研究

将一系列核心启动子区的缺失突变引入乙肝病毒（ＨＢＶ）线性转录单元,从病毒的抗原,ＲＮＡ以及子代ＤＮＡ等各个水平,分析了各缺失突变对前基因组ＲＮＡ和前核心ＲＮＡ转录的影响,对核心启动子各片段的功能进行了深入的研究。Ｃ片段缺失后检测不到ｅ抗原和前核心ＲＮＡ,却仍有核心抗原和前基因组ＲＮＡ的合成以及病毒子代ＤＮＡ的复制;而Ｂ３片段缺失后ｅ抗原和核心抗原均有显著下降,但仍能检测到两种ｍＲＮＡ的合成和病毒子     

真菌次级代谢产物11’-deoxyverticillin A（C42）通过降低自噬基因的表达抑制肝炎病毒（HBV）复制

C42属于epipolythiodioxopiperazines（ETPs）二酮呱嗪类化合物,具有多种生物学活性,包括抑制病毒复制;不过其对hepatitis B virus（HBV）复制的影响及机理鲜有报道。自噬是广泛存在于真核细胞、通过溶酶体降解长半衰期蛋白的现象,参与多种生理、病理过程。有研究发现自噬对HBV的复制至关重要。C42是否通过改变自噬来影响此病毒的复制目前还未见报道。在该研究中,我们发现表达HBV基因组的HepG2.215细胞较原始的HepG2细胞,自噬体明显增加并伴随着Akt磷酸化的增高。C42可以降低自噬基因LC3-II和p62的水平,同时会影响Akt信号通路。氯喹是一种自噬抑制剂,它的存在可以抑制C42导致的LC3-II降低,表明C42可以引起该细胞的自噬。敲降自噬基因和抑制Akt磷酸化均可以减少HBV-X蛋白表达。而利用氯喹抑制自噬体与溶酶体的融合却提高了HBV-X蛋白水平。由于HBV-X对该病毒的复制至关重要,因此,我们认为,C42通过自噬和Akt信号通路来抑制HBV的复制。     

Genetic and epigenetic alterations in hepatitis B virus-associated hepatocellular carcinoma

Hepatitis B virus(HBV) is a major cause of hepatocellular carcinoma(HCC). Its chronic infection can lead to chronic liver inflammation and the accumulation of genetic alterations to result in the oncogenic transformation of hepatocytes. HBV can also sensitize hepatocytes to oncogenic transformation by causing genetic and epigenetic changes of the host chromosomes. HBV DNA can insert into host chromosomes and recent large-scale whole-genome sequencing studies revealed recurrent HBV DNA integrations sites that may play important roles in the initiation of hepatocellular carcinogenesis. HBV can also cause epigenetic changes by altering the methylation status of cellular DNA, the post-translational modification of histones, and the expression of micro RNAs. These changes can also lead to the eventual hepatocellular transformation. These recent findings on the genetic and epigenetic alterations of the host chromosomes induced by HBV opened a new avenue for the development of novel diagnosis and treatments for HBV-induced HCC.