Characterization of human metapneumovirus from pediatric patients with acute respiratory infections in a 4-year period in Beijing, China

Background  Human metapneumovirus (hMPV) was discovered by scientists in the Netherlands as a novel respiratory virus in 2001 and had been found in children with acute respiratory tract infections (ARTI) in China. The objective of this study was to determine the importance of hMPV infection in children in Beijing and the genotypes of the circulating virus by the surveillance during a four-consecutive-year period.

Methods  Clinical specimens collected from children with ARTI from January 2006 to December 2009 were tested for hMPV by RT-PCR using primers targeting the matrix (M) gene, followed by genotyping of hMPV directly from positive samples by diplex PCR with primers for glycoprotein (G) genes. Sequence analysis was used for genotyping of those un-typable samples. Common respiratory viruses in these clinical specimens were tested by virus isolation and antigen detection, in addition to hMPV detection.

Results  Of 4730 tested specimens, 191 (4.0%) were positive for hMPV and 62.8% of 191 were identified as genotype A. The positive rate of hMPV from hospitalized patients was higher than that from outpatients each year. Most of hMPV positive children were under five years old. The peak of hMPV activity mostly occurred in late spring and overlapped with or followed that of respiratory syncytial virus (RSV) and followed by parainfluenza virus 3. Of hMPV infected cases, 68.6% were lower respiratory tract infection, among which 79.4% were hospitalized, and upper respiratory tract infection was diagnosed for 31.4% of hMPV infected children. The 9.4% of hMPV positive samples were found to co-exist with other respiratory viruses.

Conclusions  hMPV was an important pathogen for ARTI in pediatric patients, especially those under five years old. Both genotypes A and B circulated simultaneously in Beijing.

     

2012年石家庄地区急性呼吸道感染病例人偏肺病毒的分子流行病学特征

目的通过分析石家庄地区急性呼吸道感染病例中人偏肺病毒（human metapneumovirus,hMPV）的感染情况、临床特点和基因特征,掌握石家庄地区hMPV流行特点。方法采集2012年1~12月,河北省儿童医院和石家庄市人民医院两所医院门诊呼吸道感染病例标本297份,收集相关临床资料,采用多重RT-PCR方法检测包括hMPV在内的15种呼吸道病毒核酸。hMPV阳性标本通过RT-PCR扩增M基因片段,并进行核酸测序。用Mega 6软件比对序列和绘制系统进化树。结果从297份标本中检出阳性标本130份,总阳性率为43.77%,其中hMPV阳性标本5份,核酸阳性率为1.68%。患者年龄为2个月~57岁。1例病例混合感染鼻病毒和副流感病毒。临床诊断为上呼吸道感染和支气管肺炎。3株间核苷酸同源性为92.0%~99.9%。基因进化分析显示石家庄Hebei2012-1为A2b基因型,Hebei2012-2和Hebei2012-3属于B1基因型。结论石家庄地区hMPV流行基因型别为A2b和B1型。     

High seroprevalence of human metapneumovirus infection in children in Chongqing, China

Background The human metapneumovirus （hMPV） is a newly discovered respiratory viral pathogen that was first isolated in 2001 in the Netherlands. Its global distribution and long history of infection in humans have been well documented. In this study, we assessed the seropositivity of hMPV IgG antibodies in children in Chongqing, China.
Methods The specificity of the enzyme linked immunosorbent assay （ELISA） was first validated by using respiratory syncytial virus （RSV） infected, antigen subtracted reference serum and by performing western blotting using anti-hMPV animal serum. This assay was used to determine the presence of the IgG antibody against hMPV and RSV in 325 serum samples obtained from children aged 0-6 years.
Results No crossreaction was detected by ELISA between the antibodies to hMPV and RSV. The seropositivity of the anti-hMPV IgG antibody was 74.5% in children aged 0 to 5 months, 64.0% in 6 to 11 months, 72.7% in 12 to 23 months, 87.1% in 24 to 35 months and 90.3% in 3 to 6 years.
Conclusions hMPV is a common and significant respiratory pathogen in Chinese children. Almost all individuals are exposed to hMPV by age 6 years.     

Molecular and Epidemiological Characterization of Infant Botulism in Beijing,China

Laboratory-based pathogen isolation, identification, and toxicity determination were performed on samples from a suspected case of infant botulism. Mice injected with cultures generated from the enema sample and ingested Powered infant formula (PIF) presented typical signs of botulism. Antitoxins to polyvalent botulinum neurotoxins (BoNTs) and monovalent BoNT type B antitoxin had protective effects. Clostridium botulinum isolated from the enema and residual PIF samples were positive for type B toxin. Pulsed-field gel electrophoresis (PFGE) revealed that the two strains of C. botulinum isolated from the two samples produced indistinguishable pulsotypes. These findings confirmed this case of type B infant botulism associated with the ingestion of PIF contaminated by type B C. botulinum spores.     

重庆儿童医院112例毛细支气管炎病原分布特征分析

目的探讨重庆地区住院毛细支气管炎患儿鼻咽分泌物中HBoV和其他10种呼吸道病毒检出率及其流行特征变化。方法收集2006-2008年的毛细支气管炎发病高峰季节(2006年11月至2007年3月和2007年11月至2008年3月)住院毛细支气管炎患儿的鼻咽分泌物,首先通过直接免疫荧光进行呼吸道常见7种病毒筛查,然后提取鼻咽分泌物病毒的DNA和RNA,用PCR/RT-PCR的方法检测HBoV等其余10种常见呼吸道病毒相应基因片段。结果①两时间段患儿的年龄比较,差异具有显著性(P<0.01)。临床症状中,发热、呕吐、气促的发生例数差异具有显著性(P<0.05)。其余情况差异均不显著;②重庆地区住院毛细支气管炎患儿鼻咽分泌物中共检出十余种呼吸道病毒,112例患儿呼吸道标本中共检出呼吸道病毒阳性84例(75%),其中单一病毒阳性45例(40.18%),2种或2种以上病毒合并阳性39例(34.82%),2006年11月至2007年3月和2007年11月至2008年3月毛细支气管炎发病高峰季节毛细支气管炎患儿病毒检出情况不同;③112例患儿中最常见的为RSV,其余依次为PIV3,RV等,检出HBoV10例,均为合并感染。结论...     

深圳地区80例儿童下呼吸道感染人类偏肺病毒的检测

目的了解深圳地区人类偏肺病毒（hMPV）在儿童急性下呼吸道感染中的致病情况和临床特点。方法收集80例急性下呼吸道感染患儿的鼻咽分泌物，采用多重PCR扩增结合多功能流式点阵仪检测hMPV的F基因，阳性病例同时用单逆转录聚合酶链反应（RT—PCR）技术检测hMPV的核蛋白基因、N基因和L基因，并分析hMPV阳性患儿的临床特点。结果多重PCR扩增结合多功能流式点阵仪检测出hMPV 2例，其阳性率为2．5％（2／80），并经单RT—PCR证实。其他常见呼吸道RNA病毒为呼吸道合胞病毒（RSV）8例，副流感病毒（PIV）6例，鼻病毒（RhV）3例，柯萨奇及埃可病毒（CVEV）3例，流感病毒A（IFVA）4例。2例hMPV阳性患者临床过程一般。结论深圳地区儿童下呼吸道中存在有人类偏肺病毒的感染，临床上无明显特殊表现。     

小儿呼吸道人偏肺病毒感染25例流行病学和临床特征分析

目的 探讨小儿呼吸道人偏肺病毒(human metapneumovirus,hMPV)的感染阳性率、流行病学及其临床特征.方法 2002年12月至2003年12月在本科固定时间收集新入院的急性呼吸道感染患儿的鼻咽分泌物,一式两份.1份采用免疫荧光快速诊断方法用于分析包括呼吸道合胞病毒(respiratory syncytial virus,RSV)、腺病毒(adenovirus)、流感病毒(influenza virus) A、B副流感病毒(parainfluenza virus)1、2、3 这7种常见呼吸道病毒;另1份采用RT-PCR的方法检测上述患儿鼻咽分泌物中hMPV的L基因和M基因,鼻病毒与冠状病毒的基因.结果 检出 hMPV阳性标本25例,阳性率为9.7%,主要分布在秋冬季;25例hMPV阳性标本中单纯hMPV感染10例,占40%,15例(60%)系hMPV与其他病毒合并感染,其中包括8例hMPV与RSV合并感染,hMPV与鼻病毒、冠状病毒合并感染各2例,3例标本同时检出hMPV、RSV、鼻病毒为阳性;分析hMPV阳性呼吸道感染患儿的临床特征:男:女=16:9,12例(48%)临床诊断为毛细支气管炎,9例(36%)临床诊断为肺炎,4例(16%)为哮喘急性发作患儿,除hMPV和其他病毒合并感染更容易出现发热外,单纯hMPV感染与hMPV与其他病毒混合感染在发病年龄、性别、病程及临床表现上无明显差异.结论 hMPV是小儿急性呼吸道感染的重要病原,可与呼吸道其他常见病毒混合感染,仅从临床表现上很难将hMPV感染与其他呼吸道病毒感染区别.     

Current opinion on human leukocyte antigen-G in China

Since discovery and cloning of the non-classical human leukocyte antigen （HLA） class Ⅰ antigen HLA-G by Geraghty et al in 1987, a large number of studies have been carried out. HLA-G has a low polymorphism, limited distribution to normal tissues and seven isoforms resulting from its primary mRNA alternative splicing. HLA-G expression was first found on the extravillous cytotrophoblasts, at the fetal-maternal interface during normal pregnancy, which lacks the expression of HLA-A, -B and HLA Ⅱ antigens. Initial studies on HLA-G mainly addressed its function in fetal-maternal immunotolerance. Two decades later, HLA-G is now considered to be a very important immune molecule which plays a vital immune inhibitory role in the context of reproduction, oncology, transplantation, infection and also in autoimmune disease. A number of Chinese research teams are interested in, and have contributed to, the publication of more than 80 peer-reviewed articles and reviews on HLA-G over the past ten years. We summarize the key points in this field that were presented and discussed by them.     

全国人体带绦虫感染现况调查报告

目的 了解全国人体带绦虫感染现况。方法 采用《全国人体重要寄生虫病现况调查实施细则》要求,按照不同生态区和经济水平进行双重特征的分层整群随机抽样;用改良加藤厚涂片法（Kato-Katz）检查带绦虫卵。结果 在全国31个省（区、市）调查484 192人,检出带绦虫感染者1 753人,感染率0.36%;全国57个生态区中有20个生态区检出带绦虫感染者,分布在西藏等13个省（区、市）,感染率最高的生态区为藏东-川西-藏东南生态区（14.09%）。有94.69%（1 660/1 753）的感染者来自西藏,其感染率为9.25%,明显高出全国平均感染率约24.7倍。对西藏带绦虫人群分析显示：感染者多以男性、60岁以上、小学文化程度、农（牧）民为主（P<0.05）。与2004年全国调查结果（0.28%）相比,人群带绦虫感染率上升0.12%（ P<0.05）,且流行省（区、市）有所增多。结论 我国带绦虫主要流行于西藏、四川等地域的生态区,且疫情呈上升态势,建议制定有针对性的综合防控措施,控制其流行。     

An epidemiological study of resistant tuberculosis in Chongqing, China

Background The epidemiological characteristics of drug-resistant tuberculosis(DR-TB)is fundamental to improving the prevention and control of DR-TB.Mutations in katG315 is thought to be the most predictive molecule markers for Isoniazid(INH)resistance in Mycobacterium tuberculosis(MTB).However,mutations to these genes have not been thoroughly studied in China,and epidemiological evidence of their expression levels are especially lacking in the southwest of China,which has a high TB burden within the popu...     

2709例低出生体质量患儿情况分析

目的了解重庆低出生体质量患儿情况并作5年动态比较。方法回顾性分析2005年1月至2009年9月重庆医科大学附属儿童医院新生儿诊疗中心收治的2 709例低出生体质量患儿的临床资料,应用SPSS 16.0软件进行统计学分析。结果①5年来,低出生体质量患儿在同期住院新生儿中的比例始终>11%。②住院新生儿中,不同出生体质量组的患儿其平均病死率比较,差异有统计学意义(χ2=50.070,P<0.05);出生体质量在1 500~2 000 g的患儿其病死率为正常出生体质量住院患儿的1.69倍,体质量<1 500 g的患儿病死率为正常出生体质量患儿的8.79倍。③男性患儿平均出生体质量较女性患儿重(t=2.247,P<0.05)。④重庆低出生体质量患儿以早产为主,平均占87.4%。⑤初产儿平均出生体质量较经产儿(产次≥2)高(F=15.146,P<0.05)。⑥低出生体质量患儿的母亲中,45.6%曾患各类感染性疾病。⑦近50.0%的孕母在生育旺盛期以外的年龄段进行生产,40.7%的母亲处于无业状态。结论尽可能防止早产,在可控范围内减少多胎妊娠并适度控制产次等方面是重庆市低出生体质量疾控工作的重要环节,加强对低出生体质量儿,尤其是极低出生体质量儿的监测护理,提高其存活率,是降低重庆市新生儿死亡率的关键。     

Radioactivity measurements of sewerage in 4 hospitals from Chongqing, China

Monitoring of any release of radioactive materials to the environment is necessary for the environmental protection. Measurement of medical radioactive elements in the hospital sewerage is very important too. However, few study of radioactivity in hospital sewerage has been carried out or reported.     

重庆地区博尔纳病病毒种系发生分析

目的了解重庆地区博尔纳病病毒(Borna disease virus,BDV)基因特征及其种系发生。方法采用荧光定量巢式实时逆转录聚合酶链反应(fluorescence quantitative nested RT-PCR,FQ-nRT-PCR)检测健康牛、山羊、猪各50例以及病毒性脑炎(viral encephalitis,VE)患者20例外周血单核细胞(peripheral blood mononuclear cells,PBMCs)BDV p24片段,检出阳性序列和前期检出的5例重庆地区山羊和神经精神患者阳性BDV p24基因序列,与GenBank中5个国家7个动物种属33例BDV p24基因序列进行比对,分析其核苷酸和氨基酸序列同源性,重建基因系统发生树。结果10例BDV p24核苷酸序列一部分(1例VE患者)形成重庆独立支系,另一部分(7例动物和2例VE患者)汇聚至德国-瑞士-奥地利-日本-重庆混合支系。与BDV标准病毒株Strain V、H1766及He/80比对,9例核苷酸和氨基酸序列同源相似性分别为94%～100%和82%～100%,1例为95%～98%和89%～93%。结论重庆地区动物和人宿...     

人脑神经生长因子的分离和纯化及其生物学活性的鉴定

人胎脑经离心、透析和离子交换纤维素层析等方法,分离纯化２．５Ｓ神经生长因子（ＮＧＦ）。根据７ＳＮＧＦ和其活性亚单位β－ＮＧＦ的等电点的不同,分离得到２．５Ｓ的ＮＧＦ。用ＳＤＳ－聚丙烯酰胺电脉测得其分子量约１３．３。用等电聚焦电泳测得其等电点分别为９．０、９．２和９．３５。在ＰＣ１２细胞的培养过程中,加入ＮＧＦ的提取物可以促进ＰＣ１２细胞的突起生长。     

中国海域分离的啮氏艾肯菌致病性和抗生素敏感性研究

目的 就中国海域分离的啮氏艾肯菌对小鼠的毒力及伤口感染情况进行研究,并进行抗生素敏感性实验.方法 昆明小鼠腹腔注射啮氏艾肯菌悬液,观察一般状态、白细胞计数和血培养.第5天脱颈处死后解剖取脏器甲醛固定,石蜡包埋、制片,HE染色,光镜观察;昆明小鼠致伤后浸泡在含啮氏艾肯菌的人工海水中45～60 min,打捞后分笼于层流柜中饲养.于致伤浸泡后不同时间做白细胞计数及血培养,并观察一般状态和伤口炎性反应,定时取伤肢分泌物做细菌培养;实验完毕后伤肢甲醛同定,石蜡包埋、制片,HE染色,光镜检查;用K-B法做啮氏艾肯菌对各种抗生素药物敏感性实验.结果 小鼠注射啮氏艾肯菌液后,白细胞升高,感染12 h后20%小鼠心和肝脏有细菌生长.小鼠感染啮氏艾肯菌后,病变部位以肺、肝及.肾脏为重,病变性质主要是造成血液循环障碍,引起不同程度的充血、出血及血停滞现象.致伤小鼠经含啮氏艾肯菌的海水浸泡后,6、12、24、48、72 h伤肢培养均为阳性,感染后48 h时20%小鼠肠组织细菌培养阳性,其余时间点和其他组织细菌培养均为阴性.伤肢组织可见肌纤维排列疏松、紊乱、断裂,横纹肌间较多炎细胞浸润.啮氏艾肯菌对复方新诺明、哌拉西林、美洛培南、磺胺的敏感性仅为67%,对其他抗生素均表现为敏感.结论 中国海域分离的啮氏艾肯菌具有致病力,啮氏艾肯菌对复方新诺明、哌拉西林、美洛培南、磺胺的敏感性较低,对其他抗生素均表现为敏感.     

人胰岛的分离纯化和功能评价

目的 应用Ricordi人胰岛分离技术分离和纯化人的胰岛,并进行功能和安全性的评价.方法 获得6例成人尸体胰腺组织供体,采用胶原酶消化法及密度梯度离心法分离和纯化胰岛,并分析胰岛的数量、纯度和活力.采用免疫荧光法分析胰岛内分泌细胞的组成和分布;检测葡萄糖刺激的胰岛素分泌和胰岛移植后的动物血糖水平.并对分离胰岛的安全性指标进行评价.结果 分离纯化后的胰岛数量为(22.9±3.1)万IEQ,纯度为(59.0 ±8.9)%,活力为(89.0±3.0)%.免疫荧光显示,胰岛由4种内分泌细胞组成并呈正常分布.葡萄糖刺激胰岛素释放的刺激指数为8.1 ±4.0.胰岛移植于糖尿病裸鼠后,血糖在3 d后降至正常水平并维持超过30 d.分离胰岛样本的各项安全性指标在规定范围之内.结论 采用Ricordi人胰岛分离技术分离和纯化的胰岛在形态、结构、数量、纯度、活力、体内外功能以及安全性方面都达到临床胰岛移植的标准,为开展临床胰岛移植奠定了基础.     

重庆市蚊虫种类及其地理分布调查分析

目的调查研究重庆市蚊虫种类及其地理分布。方法采用人帐诱捕法、网捕法、灯诱法和人工小时法收集成蚊分类鉴定;采集蚊幼带回实验室饲养,羽化为成蚊后鉴别种类;同时检视收藏的蚊虫标本并查阅相关文献资料。结果发现重庆市有蚊虫10属73种,并对其地理分布进行了描述。结论为重庆市开展蚊虫防制和预防控制蚊媒传染病提供了科学依据。     

重庆高等医学教育现状及发展

重庆高等医学教育发展取得长足进步。在西部省市中,重庆医学院校数量和质量指标均位居前列。从全国范围来看,重庆医学教育重点指标位居中等,与浙江、广东等经济发达省份相比,还存在一定差距。四个直辖市中,北京、上海的医学教育资源丰富,与之相比,重庆的医学教育水平特别是医药院士数量和全国百强医院数量差距非常大。近期,重庆大学开始筹建医学院,同时中国科学院大学也在重庆合作建设国科大附属重庆医院,重庆应加大医药科技基础研发投入,抓住"互联网+医疗"战略机遇,汇聚医药科技人才,促进军民医学教育融合发展,提升高等医学教育整体水平。     

脑垂体生长激素的提取及其特性鉴定

本文报道一种从新鲜、冰冻猪脑垂体提取高纯度生长激素 (FGH) 的程序,包括常规的萃取、离子交换树脂层析和凝胶过滤。使用超速离心、聚丙烯酰胺凝胶电泳和生物学分析测定PGH的特性,其蛋白质的分予量是22000,沉降系数 (S20W) 为2.2267。每克新鲜猪脑垂体可获得0.8mg PGH。