湖北襄阳2016—2017年埃可病毒11型流行株基因特征分析

目的:对2016—2017年中国湖北襄阳地区分离的埃可病毒11型(Echovirus 11,Echo11)基于VP1基因进行分子进化特征分析。方法:采集2016—2017年中国湖北襄阳地区手足口病患儿肛拭子或咽拭子标本,应用real-time RT-PCR技术对肠道病毒阳性样本进行筛选,对肛拭子或咽拭子样本进行病毒分离...     

一个弱D15型家系研究

目的调查Rh血型D抗原弱表现型(弱D15)等位基因的家系遗传。方法采用常规血清学方法和PCR技术检测各家系成员RhD、C、c、E和e抗原表型,间接抗球蛋白实验确认D抗原。依据弱D15型等位基因(RHD845A)序列设计特异性引物,建立序列特异性引物-聚合酶链反应方法(sequence specificprimer-polymerase chain reaction,SSP-PCR),检测一个弱D15型先证者的4名家系成员,同时应用双管PCR技术鉴定全部家系成员的RHD基因杂合型。结果在全部4名家系成员中检出弱D15型等位基因;RHD基因杂合型结果显示4名家系成员均为RHD /RHD 纯合型。先证者的父母、外甥均有1条正常的RHD基因,为弱D15型等位基因携带者,表现为正常D阳性;先证者和姐姐各携带2条弱D15型等位基因,基因型为RHD845A/RHD845A,形成D抗原弱表现型。结论先证者为弱D15型等位基因纯合体,弱D15型等位基因为亲代遗传基因,非个体基因变异。     

巨细胞病毒感染对动脉粥样硬化的影响

目的：研究巨细胞病毒(CMV)感染和动脉粥样硬化的关系，并探讨其可能机制。方法：通过一个大样本从血清流行病学(ELISA检测患者血清中抗CMV的IgG抗体)和分子生物学(PCR检测动脉粥样硬化病变中CMV特异性基因)方面探讨巨细胞病毒感染和动脉粥样硬化的关系，并检测CMV感染对内皮细胞趋化因子表达的影响。结果：动脉粥样硬化组血清中CMV的阳性率显著高于非动脉粥样硬化组(分别为82．2％和61．0％，P=0．02)；而且动脉粥样硬化斑块中HCMV基因出现率显著高于正常血管组织(13／15和2／7，P=0．01)；CMV感染还可上调内皮细胞：ECV-304表达MCP-1。结论：CMV感染参与动脉粥样硬化的形成和发生，这可能与CMV上调内皮细胞趋化因子表达有关。     

Genetic variability in the sapovirus capsid protein

Sapovirus (SV), which causes gastroenteritis in humans, is composed of genetically divergent viruses classified into 5 genogroups. In this study, 2.2-kb nucleotide sequences of the 3′ terminus of the genome of 15 SV strains detected in Japan were determined. The 15 SV strains could be classified into four genogroups (GI, GII, GIV and GV), and in two of these, GI and GII, 10 genotypes were identified. The amino acid sequences of the central variable region of the capsid protein showed less than 81% identity when strains belonging to different genotypes were compared. It was therefore supposed that antigenic variety exists between different genotypes. These results will be useful for further genetic and antigenic analyses of SV.     

人巨细胞病毒在广西不同人群中的感染情况及其与肾脏疾病关系的初步研究

目的 调查广西壮族自治区孕妇、患病新生儿、肾病患者、肾移植患者及正常体检者尿中人巨细胞病毒阳性率,了解人类巨细胞病毒在广西不同人群中的分布情况,初步探讨人巨细胞病毒感染与肾脏疾病的关系.方法 用荧光定量PCR检测研究对象晨尿中人巨细胞病毒DNA.结果 孕妇、患病新生儿、肾病者、肾移植患者及正常体检者尿中人巨细胞病毒DNA阳性率依次为8.18%、3.45%、18.54%、25.42%、0.56%,其在各类肾病患者中的感染率均在10%左右,尤其在狼疮肾炎患者中的感染率更是达到27.78%.结论 人巨细胞病毒在黄疸和早产新生儿中的感染率均较高,需要加强母婴人巨细胞病毒感染的防控工作.人巨细胞病毒很可能在肾病患者发病时对患者肾脏造成损害,并且加重肾病患者的病情,成为肾衰的一个诱因.     

Characterization of a multigene family encoding an endopolygalacturonase in Sclerotinia sclerotiorum

Sclerotinia sclerotiorum produces several polygalacturonases which together with other pectinolytic enzymes are involved in the degradation of pectin. A number of different genomic clones were isolated by screening a genomic DNA library in phage EMBL3. Southern-blot and restriction mapping indicate that seven genes constitute two subfamilies of a multigene family encoding endopolygalactutonase. Using pulsed-field gel electrophoresis to separate S. sclerotiorum chromosomes each subfamily was found to hybridize to a different chromosome. A comparison of the nucleotide sequence for the coding region of three members of the gene family reveals surprisingly few base substitutions suggesting that this gene family arose from recent multiple duplication events.     

乙型肝炎病毒D基因型系统进化树分析

目的：研究HBV D基因型不同毒株全基因进化关系。方法：用聚合酶链式反应（PCR）、克隆及核酸序列测定的方法，测定了1例中国人慢性无症状携带者感染的乙型肝炎病毒D基因型全基因序列。此D基因型毒株全基因序列GenBank Accession为AF280817，将GenBank中已发表的HBV D基因型30株的全序列进行了系统进化树分析。结果：中国株HBV D基因型与源于瑞典（Sweden）的4株HBV D基因型全基因的进化距离最近；地中海地区及欧洲国家系HBV D基因型分布的优势地域，亚洲并非HBV D基因型分布的优势地域。结论：HBV D基因型病毒株传入来源、迁移的方向不同以及病毒株在具有不同遗传和免疫特质的宿主中的长期选择是形成HBV D基因序列病毒进化差异的原因。     

人3型腺病毒广州分离株基因组的进化分析

目的研究人3型腺病毒(human adenovirus type 3,HAdV-3)全基因组序列的基本特征并进行基因组的进化分析。方法取患儿鼻咽分泌物进行病毒分离,用中和试验和PCR进一步鉴定分型;克隆酶切片段并测序,利用软件对腺病毒基因组蛋白进行进化树分析。结果分离得到2株HAdV-3,基因组全长分别为35273bp和35269bp,2株均含有39个DNA编码序列和2个RNA编码序列。非编码区均保守,存在回文序列和反转重复序列。进化分析表明HAdV-3及其他B组腺病毒与猿腺病毒21型具有更近的进化关系。结论获得广州地区分离的2株HAdV-3,基因组全长35273bp和35269bp;人B组腺病毒与猿腺病毒存在一定的进化关系。     

我国分离的两株病毒为重组甲病毒

目的 明确我国分离的XJ－90260和XJ－91006病毒的分类地位、种系发生和遗传型。方法 特异引物逆转录－聚合酶链反应（RT－PCR）扩增两株病毒的NSP4、E1基因区和3′端非编码区,测序,进行核苷酸序列同源性比较和3′端非编码区核苷酸序列分析,并结合同属其他病毒这些基因区的核苷酸序列进行种系进化分析。结果 两株病毒的核苷酸序列同源性为100％,与西方马脑炎病毒的核种酸序列同源性最高,具有西方马脑炎病毒3′端非编码区结构特征,其NSP4基因区与东方马脑炎病毒同源,E1基因区与辛德毕斯病毒同源,两株病毒均位于西方马脑炎病毒B组,与西方马脑炎病毒俄罗斯分离株进化关系最近。结论 我国分离的XJ－90260和91006病毒属于西方马脑炎病毒同一遗传型,均为重组甲病毒。     

The gene for Best''s macular dystrophy is located at 11q13 in a Swedish family

A large Swedish family with more than 250 cases of Best's macular dystrophy has been clinically and genetically studied. The gene was traced to a couple born in central Sweden in the 17th century. Highly significant evidence for genetic linkage to DNA markers on chromosome 11q13 was detected. A lod score of 15.12 was obtained at recombination fraction 0.01 with DNA marker INT2 (also called FGF3). The retinally expressed gene ROM1, which maps to the same chromosomal region is a candidate for this genetic disease.     

Serostatus of echovirus 11, coxsackievirus B3 and enterovirus D68 in cord blood: The implication of severe newborn enterovirus infection

BackgroundMaternal transplacental antibody is an important origins of passive immunity against neonatal enterovirus infection. Echovirus 11 (E11) and coxsackievirus B3 (CVB3) are important types causing neonatal infections. There were few investigations of enterovirus D68 (EVD68) infection in neonates. We aimed to investigate the serostatus of cord blood for these three enteroviruses and evaluate the factors associated with seropositivity.MethodsWe enrolled 222 parturient (gestational age 34–42 weeks) women aged 20–46 years old between January and October 2021. All participants underwent questionnaire investigation and we collected the cord blood to measure the neutralization antibodies against E11, CVB3 and EVD68.ResultsThe cord blood seropositive rates were 18% (41/222), 60% (134/232) and 95% (211/222) for E11, CVB3 and EVD68, respectively (p < 0.001). Geometric mean titers were 3.3 (95% CI 2.9–3.8) for E11, 15.9 (95% CI 12.5–20.3) for CVB3 and 109.9 (95% CI 92.4–131.6) for EVD68. Younger parturient age (33.8 ± 3.6 versus 35.2 ± 4.4, p = 0.04) was related to E11 seropositivity. Neonatal sex, gestational age and birth body weight were not significantly different between the seropositive group and the seronegative group.ConclusionCord blood seropositive rate and geometric mean titer of E11 were very low, so a large proportion of newborns are susceptible to E11. The circulation of E11 was low after 2019 in Taiwan. A large cohort of immune naïve newborns existed currently due to lack of protective maternal antibodies. It is imminent to monitor the epidemiology of neonates with enterovirus infections and strengthen the relevant preventive policies.     

Linkage analysis between manic-depressive illness and markers on the long arm of chromosome 11

The long arm of chromosome 11 is one of the most interesting regions in the search for major genes involved in the etiology of manic-depressive illness. Several candidate genes have been identified, including the gene encoding the dopamine D2 receptor, the M1 muscarinic receptor, and porfobillinogen deaminase. Furthermore, different families with co-segregation of psychiatric illness and structural chromosome abnormalities involving regions 11q21, 11q22.3, and 11q25 have been reported. Using narrow as well as broad phenotypic models, conservative genetic parameters, models with dominant or recessive modes of inheritance, and various methods to reduce misclassification, the present study did not find evidence for a major gene causing manic-depressive illness on the long arm of chromosome 11. In the broader phenotypic models multi-point analyses excluded at least 11q14 to 11q23.3, approximately 60 cM, even in one large family. Assuming homogeneity close linkage to DRD2 was excluded for all dominant models, and also in the affecteds-only analyses in the large family alone. © 1995 Wiley-Liss, Inc.     

Phosphorylation of the RNA polymerase II carboxyl-terminal domain in human cytomegalovirus-infected cells and in vitro by the viral UL97 protein kinase

The carboxyl-terminal domain (CTD) of the largest subunit of RNA polymerase II (RNAP II) ordinarily exists in electrophoretically distinct hypophosphorylated and hyperphosphorylated forms. Human cytomegalovirus infection induced forms of this subunit whose electrophoretic mobilities were intermediate without decreases in abundance of the original forms. Phosphatase treatment nearly eliminated the intermediate migrating forms. In vitro, the viral protein kinase, UL97, phosphorylated this subunit, a recombinant protein containing the CTD, and peptides containing the CTD consensus sequence, YSPTSPS. Phosphorylation occurred predominantly on serine 5 and was substantially reduced when either serine 2 or 5 was already phosphorylated. The abundance of the intermediate and hypophosphorylated forms was reduced at most twofold during infections in which UL97 was genetically or pharmacologically inhibited. These results identify a new pattern of RNA polymerase II modification induced by virus infection and a viral enzyme that phosphorylates the CTD in vitro, but only possibly in vivo.     

Molecular phylogenetic and positive selection analysis of Japanese encephalitis virus strains isolated from pigs in China

Japanese encephalitis virus (JEV) is one of the most important virus which causes encephalitis. This disease is most prevalent in the south, southeast and the east region of Asia. In this study, two JEV strains, named JEV/SW/GD/01/2009 and JEV/SW/GZ/09/2004, were isolated from aborted fetuses and seminal fluid of pigs in China. To determine the characteristic of these virus isolates, the virulence of two newly JEV isolates was investigated, the result evidenced that the JEV/SW/GD/01/2009 did not kill mice, while the JEV/SW/GZ/09/2004 displayed neurovirulence with 0.925 log₁₀ p.f.u./LD50. Additionally, the full genome sequences of JEV were determined and compared with other known JEV strains. Results demonstrated that the genome of two JEV isolates was 10,976 nucleotides (nt) in length. As compared to the Chinese vaccine strain SA14-14-2, the JEV/SW/GD/01/2009 and the JEV/SW/GZ/09/2004 showed 99.7% and 97.5% identity at the nucleotide level, 99.6% and 96.7% identity at the amino acid level, respectively. Phylogenetic analysis, based on the full-length genome revealed that two JEV isolates were all clustered into genotype III compared to the reference strains. Furthermore, selection analyses revealed that dominant selective pressure acting on the JEV genome was purifying selection. Four sites under positive selection were identified: codon 521 (amino acid E-227), 2296 (amino acid NS4b-24), 3048 (amino acid NS5-521) and 3055 (amino acid NS5-528). Amino acid E-227 was proved to be related to neurovirulence. Taken together, the molecular epidemiology and functional of positively selected amino acid sites of two newly JEV isolates were fully understood, which might be helpful to predict possible changes in virulence.     

Epigenetic mutations in 11p15 in Silver-Russell syndrome are restricted to the telomeric imprinting domain

Introduction

Silver‐Russell syndrome (SRS; also know as Russell‐Silver syndrome) is a heterogeneous syndrome which is characterised by severe intrauterine and postnatal growth retardation and typical dysmorphic features. Recently, the first SRS patients with (epi)genetic mutations in 11p15 affecting the telomeric imprinting domain have been identified. Interestingly, opposite mutations are associated with Beckwith‐Wiedemann syndrome (BWS). However, the general significance of epigenetic mutations in 11p15 for the aetiology of SRS remained unclear.

Methods

We screened a cohort of 51 SRS patients for epimutations in ICR1 and KCNQ1OT1 by methylation sensitive Southern blot analyses.

Results

ICR1 demethylation could be observed in 16 of the 51 SRS patients, corresponding to a frequency of approximately 31%. Changes in methylation at the KCNQ1OT1 locus were not detected.

Discussion

Combining these data with those on maternal duplications in 11p15, nearly 35% of SRS cases are associated with detectable (epi)genetic disturbances in 11p15. We now have to also consider a general involvement of 11p15 alterations in growth retarded patients with only minor or without further dysmorphic features. SRS and BWS may now be regarded as two diseases caused by opposite (epi)genetic disturbances of the same chromosomal region displaying opposite clinical pictures.     

Identification of a novel STK11 gene mutation in a family affected with hereditary Peutz-Jeghers syndromeCSCD

Objective: To explore the genetic basis for a family affected with Peutz-Jeghers syndrome (PJS). Methods: Genomic DNA was extracted from peripheral blood and oral swab samples from the patient and her relatives. Next-generation sequencing (NGS) was used to analyze 106 target genes by capturing the exons and adjacent intronic regions. Suspected pathogenic mutation was verified by NGS. Results: A missense STK11 mutation was detected in the proband, which was not reported previously. The mutation has caused substitution of Leucine by Proline. NGS has detected the same mutation in the mother but not among other relatives. Conclusion: This hereditary case of PJS may be attributed to the missense mutation of the STK11 gene. © 2018 West China University of Medical Sciences. All rights reserved.     

2006年北京市外来人口中HIV-1感染者流行毒株gag基因序列测定和亚型分析

目的了解北京市外来人口中HIV-1亚型的特点和流行规律。方法随机采集北京市2006年外来人口中新确证HIV-1感染者的抗凝全血标本80份，分离血浆，提取病毒RNA，用套式聚合酶链反应扩增病毒gag基因，并进行序列测定和亚型分析。结果系统进化分析确定北京市外来人口HIV-1毒株属于8个亚型，分别为B亚型4份，泰国B亚型15份，C亚型1份，CRF01-AE亚型5份，CRF02-AG亚型1份，CRF07-BC亚型29份，CRF08-BC亚型3份，CRFl5—01B亚型1份。结论北京市外来人口中己存在8种HIV-1亚型和流行重组型，应该加强对HIV-1亚型变异的监测。