经母婴传播获得乙型肝炎病毒(HBV)感染儿童及其母亲体内HBV前S/S基因变异特点研究

目的 通过对经母婴传播获得HBV感染的子女及其母亲慢性携带者体内HBV前S／S序列研究,了解不同程度病毒血症下,来源相同HBV变异特点。方法根据HBV病毒血症高低分为3组,每组5对母子。应用T-A克隆技术构建重组质粒pGEM-前S／S、双酶切鉴定并分析。结果 每对母子HBV亚型相同,各组5对母子中4对为B／adw2、1对为C／adrq 亚型。每组中B／adw2亚型HBV分析显示：高病毒血症组间或低病毒血症组间HBV前S／S基因变异数目及位点差异均无显著性,低病毒血症病人变异数目及位点明显高于高病毒血症病人。两低病毒血症组的13／16个克隆存在较多位点变异,每个克隆86—94个;11／13克隆的86个、2／13克隆的90个核苷酸变异相同,分别引起37、38个氨基酸改变、大多位于免疫表位内或附近;这两个变异序列有62个核苷酸变异位点相同、可致28个氨基酸变异。变异与年龄无关。结论 HBV变异可能与感染的时间长短无关;抗-HBe阳性的低病毒血症病人体内HBV前S／S出现较多的变异,其变异是有规律的,可能与病毒逃避免疫攻击有关。     

乙型肝炎病毒S基因变异、基因型与宫内感染免疫失败的关系

目的 探讨HBV S基因变异、基因型与宫内感染免疫失败的关系.方法选择东南大学附属南京第二医院出生的35例宫内感染免疫失败的幼儿及其母亲,实时荧光定量PCR法检测血清HBV DNA含量;并扩增其HBV S基因序列,测序并通过DNASTAR软件与基因库标准序列比对.结果幼儿及其母亲HBV DNA定量检测结果均＞1×106拷贝/mL;幼儿及母亲HBV S基因核苷酸变异率分别为11.4%和17.1%;母婴序列同源性＞99.3%;35对母婴中,23对HBV基因型为C型,血清型为adr亚型,12对为B型,血清型为adw亚型,母婴基因型相同.结论 HBV S基因是否变异可能不是高病毒血症患者宫内感染免疫失败的主要因素;基因分型并不能预测和评价新生儿是否发生宫内感染和免疫失败.     

乙型肝炎病毒前S基因变异与乙型肝炎相关晚期肝病关联性的临床研究

目的研究乙型肝炎病毒(HBV)前S基因变异与乙型肝炎相关晚期肝病及慢加急性肝衰竭(ACLF)的关系。方法收集慢性乙型肝炎(CHB)、乙型肝炎相关性肝硬化(LC)、乙型肝炎相关性肝细胞癌(HCC)患者共71例的血标本及临床资料,提取血清HBV DNA,进行基因分型,PCR扩增HBV DNA前S基因,所得阳性PCR产物进行测序,分析测序结果。结果检测样本中基因型C49例,基因型B21例,混合基因型B+C1例,C为优势基因型。PreS变异率在CHB(不包括ACLF)、LC、HCC三组中分别为18.75%、48.78%、66.67%,CHB组PreS区变异率低于LC组(P0.05)和HCC组(P0.05)。ACLF组PreS区变异率(80%)明显高于CHB组(18.75%)(P0.005)。HBV DNA≥104拷贝/mL在发生PreS区变异组为30例(90.9%),未发生PreS区变异组为19例(50.0%),两组差异有显著统计学意义(P0.001)。Logistic回归分析示HBV DNA≥104拷贝/mL、HBV相关晚期肝病(LC、HCC)与PreS区变异相关(OR值:14.153,95%CI:3.412~58.701;OR值:3.924,95%CI:1.178~13.074)。结论 HBV前S区变异与乙型肝炎相关晚期肝病及肝衰竭的进展有关。     

武汉地区儿童乙型肝炎患者病毒S基因"a"决定簇变异的研究

目的研究武汉地区儿童乙型肝炎患者中乙型肝炎病毒（HBV）S基因变异株的分子流行病学特征。方法通过流行病学调查筛选出30例经乙型肝炎疫苗免疫的儿童乙型肝炎患者及30例未免疫儿童乙型肝炎患者，利用聚合酶链反应（PCR）扩增HBV S基因片段，直接将PCR产物进行DNA序列测定。结果在60例儿童乙型肝炎患者中检测出55例adw血清亚型，其中8例HBV S基因发生氨基酸置换；直接测序检测的已免疫和未免疫儿童乙型肝炎患者S基因氨基酸置换频率分别为20．0％和6．7％，感染基因变异株的患儿发病年龄明显偏大，其疫苗免疫年限均较长。结论本地区流行的乙型肝炎病毒毒株主要为adw血清亚型；乙型肝炎疫苗具有免疫选择表面抗原基因变异株的作用；基因变异株病毒感染有随疫苗免疫年限延长而明显增加的趋势。     

乙肝病毒基因型与肝脏病理改变的关系

目的:探讨乙型肝炎病毒基因型与慢性乙型肝炎患者肝脏病理变化的关系.方法:应用乙肝病毒型特异性引物采用巢式聚合酶链反应(PCR)和荧光定量聚合酶链反应(FQ-PCR),对北京佑安医院住院92例慢性乙型肝炎患者进行乙型肝炎病毒基因型及亚型分析,参照2000年《病毒性肝炎防治方案》对慢性肝炎进行病理分级、分期诊断.结果:92例慢性乙型肝炎患者中HBV基因型分布为B型17例(B2亚型),B/C混合型17例(B2/Ca亚型),C型58例(Ca亚型).17例HBV B型感染患者中病理诊断肝脏炎症活动分级为G1-G3期分别为35.29%,58.82%,5.88%;肝脏纤维化程度分级为S1-3级分别为58.82%,29.41%,11.76%,17例HBV B/C型感染患者中病理诊断肝脏炎症活动分级为G1-G3期35.29%,52.94%,11.76%;肝脏纤维化程度分级为S1-3级23.52%,52.94%,23.52%,58例HBV C型感染患者中病理诊断肝脏炎症活动分级为G1-G4期31.03%,24.14%,36.21%,8.62%;肝脏纤维化程度分级为S1-4级25.86%,39.66%,5.17%,29.31%;HBV三组不同基因型的慢性乙型肝炎患者肝组织病理检查有统计学意义(x~2=15.13,P<0.01).HBV B型与B/C型感染患者年龄在21-30岁组58%-76%,31-40岁组17.6%-29.4%,C型感染患者年龄在21-30岁组25%,31-40岁组46.6%,40岁以上有24.24%;不同HBV基因型感染患者的年龄分布有显著性差异(x~2=9.54,P<0.05).结论:慢性乙型肝炎患者HBV基因型中C型比例明显高于B型与B/C型.HBV C型患者肝脏病理变化较B型与B/C型严重.不同HBV基因型感染患者的年龄分布不同.     

乙型肝炎病毒S基因变异与临床

变异是生物界普遍存在的现象，是物种适应生存压力的结果。乙型肝炎病毒(hepatitis B virus，HBV)与其它DNA病毒相比具有很高的变异率。随着研究的深入，表明乙型肝炎病毒S基因变异与乙肝疫苗预防接种失败、HBIG预防移植后肝脏再感染、HBsAg(-)的HBV感染及疾病严重性等均有密切关系。本文就HBV S基因变异及其在临床中的意义作一综述。     

未经治疗的慢性乙型肝炎患者乙型肝炎病毒耐药变异、基因型和血清型研究

目的 研究未经核苷(酸)类似物(NA)治疗的慢性乙型肝炎患者HBV耐药变异、基因型、基因亚型和血清型特点.方法 从北京大学附属医院收集97例未经NA治疗的慢性乙型肝炎患者血清,用半巢式聚合酶链反应-直接测序法获得HBV全长逆转录酶区序列,用生物信息学技术筛查该区内11个经典耐药变异位点并鉴定基因型、基因亚型和血清型.用统计分析软件SPSS11.0进行t检验和χ~2检验. 结果 HBV在11个经典耐药变异位点上均为野生型氨基酸;B基因型和C基因型分别占36.1%(35/97)和63.9%(62/97),前者均属B2亚型,后者C2亚型占91.9%(57/62),C1亚型占6.5%(4/62),1例未能分出亚型.已知出生地的患者中,71.9%(23/32) B基因型感染者出生于我国南方地区,81.6%(40/49) C基因型感染者出生于北方地区,基因型地域分布特点明显,χ~2=23.19,P<0.01.血清型为adr者占60.8%(59/97),与C基因型相关;为adw者占38.1%(37/97),与B基因型相关,χ~2=87.83,P<0.01.结论 未经NA治疗的慢性乙型肝炎患者体内野毒株为优势株,其基因型、基因亚型和血清型与患者出生地有关.     

慢性乙型肝炎患者HBV基因型和亚型分布调查

目的研究慢性乙型肝炎患者HBV基因型和亚型流行情况。方法应用HBV基因型和亚型特异性引物PCR法对北京、长春、大连、西安、石家庄、郑州和合肥7个城市660份HBVDNA阳性慢性乙型肝炎患者血清进行基因型和亚型分析。结果在660份HBVDNA阳性血清中,B基因型、C基因型和B/C混合感染分别为16.67%(110/660)、74.54%(492/660)和8.79%(58/660);在C基因型中,C1亚型6例(1.22%)、C2亚型473例(96.14%)、C1/C2混合基因亚型13例(2.64%);B基因型均为Ba亚型,B基因型和C基因型混合感染者均为Ba与C2亚型混合感染,未发现其他基因型和基因亚型;不同基因型感染患者HBeAg阳性率差异无统计学意义(P=0.153);B基因型和C基因型患者之间血清HBVDNA水平差异无统计学意义(6.37±1.62lgcopies/ml对6.29±1.76lgcopies/ml),但均高于B和C基因型混合感染患者(5.25±1.65lgcopies/ml)。结论这7个城市慢性乙型肝炎患者以B基因型和C基因型感染为主,有部分B/C基因型混合感染。HBV亚型以Ba和C2亚型占优势。     

HBsAg/HBsAb双阳性慢性乙型肝炎患者HBV S基因免疫逃逸相关变异分析

目的分析HBsAg/HBsAb双阳性慢性乙型肝炎患者HBV S基因主要亲水区免疫逃逸相关位点变异特点。方法收集89例HBsAg/HBsAb双阳性和148例HBsAg单阳性的慢性乙型肝炎患者血清及临床资料,提取患者血清HBV DNA,扩增患者HBV S基因并进行测序,应用DNASTAR Lasergene Meg Align软件对主要亲水区已有文献报道的46个免疫逃逸相关位点及新增N-糖基化变异进行比对分析。结果 2组患者在年龄、ALT、TBIL、HBV DNA载量和HBe Ag阳性率方面差异均无统计学意义(P均0.05)。HBsAg/HBsAb双阳性患者HBV S基因主要亲水区变异的总检出率为31.46%,明显高于HBsAg单阳性患者的18.92%(P0.05),其中s L110I/S、s T113N/S、s T131I/N/P和s S143L/M/T的变异检出率明显高于单阳性组;双阳性患者多位点联合变异检出率亦明显高于单阳性患者(20.22%vs.6.08%,P0.05)。双阳性患者新增N-糖基化变异检出率高于单阳性患者(7.87%vs.2.03%),差异具有统计学意义(P0.05)。结论 HBsAg/HBsAb双阳性患者比HBsAg单阳性患者的HBV S基因免疫逃逸相关变异种类更多,单位点和多位点联合变异检出率更高,并且新增N-糖基化变异检出率也更高,这些变异可能是引起慢性乙型肝炎患者HBsAg/HBsAb双阳性共存的驱动因素之一。     

急性乙型肝炎患者前S1抗原与前S1抗体检测的临床意义

目的 分析急性乙型肝炎(acute hepatitis B,AHB)患者前S1 抗原(PreS1Ag)和前S1 抗体(抗PreS1)与HBV DNA转阴、HBeAg 和HBsAg 转阴/血清学转换的关系,初步探讨PreS1Ag/抗PreS1 对AHB 预后的预测价值.方法 纳入15 例AHB患者,动态抽取血清标本,检测...     

Selective transmission of variant genomes from mother to infant in neonatal fulminant hepatitis B

Hepatitis B virus (HBV) nucleotide sequences isolated from mother/child pairs were analyzed in three cases of neonatal fulminant hepatitis B (FHB). Heterogeneous HBV sequences consistent with both adw2 and ayw subtype were found in all three mothers. In one case, in which the child survived, both subtypes were transmitted. By contrast, only the ayw subtype was present in the two other children with a fatal course of FHB. In one fatal case, studied in greater detail, multiple HBV variants (viral quasi-species) were identified in both mother and child. A direct sequence comparison showed that only a subfraction of the virus pool from the mother was transmitted and that multiple new mutations emerged in the child. These data suggest that a minor HBV subpopulation from the mother may prevail as the dominant species in the child and that neonatal FHB is associated with the selection of mutant strains.     

A Quasi species of the pre-S/S gene and mutations of enhancer II/core promoter/pre-C in mothers and their children infected with hepatitis B virus via mother-to-infant transmission

BACKGROUND: Hepatitis B virus (HBV) infection has a tendency to be chronic. The quasi species of HBV in the pre-S/S gene and mutations in enhancer II (EnhII)/core promoter (CP)/pre-C of HBV were studied in asymptomatic carrier (AsC) mothers and their children with different virus loads, to gain a better understanding of the pathogenic mechanisms of HBV. METHODS: Quasi species were analyzed using an established phylogenetic tree based on the sequences of the pre-S/S gene from 3 mother-child pairs. The mutations of EnhII/CP/pre-C were studied in 15 mother-child pairs. RESULTS: Substitution was the main mutation model. The phylogenetic tree indicated that the pre-S/S gene in every mother-child pair had the same root. The sequences of the pre-S/S gene of each patient were somewhat different from one another, but there was limited evolutionary distance between them. The evolutionary distances between the pre-S/S gene and the base of the tree in patients with low virus loads were greater than those in patients with high virus loads. The mutations in patients with low virus loads were much more frequent than those in patients with high virus loads and were not related to age, irrespective of whether the pre-S/S gene or EnhII/CP/pre-C was considered. CONCLUSIONS: There are HBV quasi species in the sera of AsCs, and the mutations are related to virus load and hepatitis B e antigen seroconversion, irrespective of age.     

A Pilot Study of Mother-Child Interactions in Children with Bronchial Asthma

The mother-child interactions in children with bronchial asthma were studied in five pairs. Each mother-child pair was videotaped in two 42-minute play-sessions. Detailed four-second interval analysis of behaviour patterns in each interacting dyad was obtained. Computer-analyzed behaviour samples were compared with those obtained from a control group of eight nonasthmatic children and their mothers. All children involved in the study ranged between the ages of 1-3/4 and 4 years. A significant difference was found in the ambivalent status variable in the asthmatic children's group as compared with the control group.

The extreme interactional patterns were identified in the asthma pairs. A series of eight weekly instruction sessions provided the mother an opportunity to correct some of her behaviours while engaged in playing with her child. Pre-and post-instructional analysis of the interactions between mother and asthmatic child provided a measure of any significant behaviour changes. The method of instructing the mother to modify her behaviour via a radio intercommunication device while she was playing with her child was shown to be an effective therapeutic tool.     

Hepatitis B virus infections in families in which the mothers are negative but the fathers are positive for HBsAg.

We studied a total of 37 families, in which HBsAg was positive in either or both of father and mother, to assess intra-familial transmission of hepatitis B virus (HBV). The HBsAg positive rate for children with HBsAg-negative mothers was significantly lower than that with positive mothers (4 of 31, 12.9% versus 18 of 32, 56.3%, p<0.01) of course. However, there were three families in which the infection source for children was thought to be fathers, not mothers, i.e., of eight children in these three families with HBsAg +/- father/mother pairs, 4 (50%) were positive for both HBsAg and HBV DNA of genotypes identical to those of their fathers, and another child was positive for HBcAb despite being negative for HBsAg. Interestingly, moreover, all the mothers in these three families were HBcAb-positive even though HBsAg-negative, suggesting that not only father-to-child but also inter-spouse HBV transmission might have occurred. With these findings we would suggest that all the family members with HBsAg-positive fathers should receive HBV vaccine, let alone for those with HBsAg-positive mothers.     

A Pilot Study of Mother-Child Interactions in Children with Bronchial Asthma

The mother-child interactions in children with bronchial asthma were studied in five pairs. Each mother-child pair was videotaped in two 42-minute play-sessions. Detailed four-second interval analysis of behaviour patterns in each interacting dyad was obtained. Computer-analyzed behaviour samples were compared with those obtained from a control group of eight nonasthmatic children and their mothers. All children involved in the study ranged between the ages of 1-3/4 and 4 years. A significant difference was found in the ambivalent status variable in the asthmatic children's group as compared with the control group.

The extreme interactional patterns were identified in the asthma pairs. A series of eight weekly instruction sessions provided the mother an opportunity to correct some of her behaviours while engaged in playing with her child. Pre-and post-instructional analysis of the interactions between mother and asthmatic child provided a measure of any significant behaviour changes. The method of instructing the mother to modify her behaviour via a radio intercommunication device while she was playing with her child was shown to be an effective therapeutic tool.     

Molecular evidence for transmission of GB virus-C/Hepatitis G virus infection within family: close relationship between mother and child

BACKGROUND/AIMS: GBV-C/Hepatitis G virus (HGV) is transmitted parentally, but some of the transmitted routes are still unclear. This study was conducted to investigate the relationship of family members in the transmission of HGV. METHODOLOGY: The study group enrolled 11 children from four index mothers with HGV viremia, whereas the control group had 14 children from seven matched mothers without HGV infection. Detection of circulating HGV RNA and anti-E2 antibody was conducted. Comparison of partially HGV nucleotide sequences at the 5' non-coding and non-structure 5 regions was performed among the infected family members. RESULTS: There was higher HGV viremia in children from the index mothers with HGV infection than those from mothers in control (3/11 vs. 0/14, p < 0.05). Nucleotide sequences analysis showed higher similarity between mothers and their children in the same family (98.6% to 100%) than between mothers of different families (87.8% to 92.4%). The phylogenetic construction also demonstrated distinct sequence clusters for mother and child/children of each family. CONCLUSIONS: The relationship of HGV transmission between mother and children was closely related in family. Spread of HGV might occur intrafamilially.     

The Complement-Activating Capacity of Maternal IgG Antibodies to Blood Group A in Paired Mother/Child Serum Samples

Background and objectives : The aim of this study was to investigate whether IgG antibodies to blood group A bind or activate complement after crossing the placenta. Materials and methods : IgG anti-A concentrations as well as C1-q-binding and the capacity of anti-A to form the C5b-9 membrane attack complex were measured in sera of 61 mother/child pairs. The enzyme-linked immunosorbent assay was used to quantitate anti-A or anti-B in maternal and neonatal serum samples. Results : The concentrations of IgG anti-A in ABO-identical and ABO-compatible children born to type O or B mothers correlated significantly with maternal concentrations (p < 0.05), whereas type A or AB children had strikingly diminished concentrations of IgG anti-A. The IgG anti-A concentrations were analyzed in two different groups: first, 44 mother/child pairs with mothers and children group O or B (group I), and second, 17 pairs with mothers group O or B and children group A or AB (group II). In group I, identical or compatible pairs with regard to IgG anti-A, a significant correlation was found by Spearman's rank analysis, whereas the same analysis revealed no significant correlation in group II. The capacity of anti-A to activate complement was also more marked in newborns who were ABO-identical or ABO-compatible with their mothers, and this capacity was virtually absent in cord blood samples from incompatible babies. Conclusions : Tests for complement-dependent or complement-independent ABO blood group antibodies must be interpreted in the light of the blood group of the child.     

Single‐nucleotide substitution of Hepatitis B virus in intrauterine infection

The relationship between hepatitis B virus (HBV) gene polymorphism and intrauterine infection has not been completely illuminated. Six pairs of mother and infant from intrauterine infection group and six mothers from nonintrauterine infection group in the previous study were randomly selected and separately divided into group M (Mother group), group N (Neonate group) and group NM (Negative‐mother group) in this study. We found that age, gestational weeks, HBsAg titre, HBeAg titre and HBV DNA level of mothers from group M and group NM were not significantly different. Pre‐S1/S2 and S regions in HBV genome were amplified, inserted into pUC19 plasmid and sequenced. It was found that all clone sequences clustered into genotype C ( AY123041 ) through the Genotyping tool in NCBI and phylogenetic trees. Compared with AY123041 , there were 20 (11 plus 9) mutations significantly different in the three groups. Most of the mutations were synonymous in pre‐S1/S2/S region, while mutations of C2990T, T3205A, A167G, C407A, A667T and A680C resulted in amino acid substitution of A90V, S162T, T47A, P127T, L213F and I218L, respectively. In addition, most of the 20 mutations caused amino acid substitution in polymerase region for the tight structure of HBV genome. The occurrence and location of mutations indicated that mutation of C2990T only existing in group NM may serve as an index for nonintrauterine infection. In contrast, the incidence of intrauterine HBV infection from mothers with mutation of T3205A was lower. Then, mutations of G403A, T670G, A673G, A167G, C407A, A667T and A680C may be closely related to intrauterine HBV infection.