2010-2012年广州地区人冠状病毒及其亚型分布

目的了解2010—2012年广州地区发热呼吸道感染病人的冠状病毒（HCoV）流行状况和特点,为进一步了解人冠状病毒的流行规律及防控奠定基础。方法收集广州市2010—2012年5家哨点医院共3376例符合要求的发热同时伴随有呼吸道感染症状病人的咽拭子标本及相关临床信息,采用Real—timePCR方法检测冠状病毒的核酸并对其进行主要型别鉴定,从人群分布、月份分布及各型别发病率等方面进行分析。结果3376例发热伴有呼吸道感染症状的病例中,55例为冠状病毒阳性标本,检出率为1．63％。亚型0C43、229E和NL63阳性的标本分别为23、13和19例,阳性率分别为0．68％、0．39％和0．56％,其中SARS、HUKl及EMC亚型未检出。冠状病毒流行高峰为1月（5．33％）,不同月份之间发病率比较差异有统计学意义（xz=38．197,P〈0．01）;阳性率最高年龄组为60。93岁,检出率为2．6％。亚型OC43发病高峰为12—1月,与其他月份比较差异有统计学意义（xL30．054,P〈0．01）,检出率最高的年龄组为60～93岁（1．3％）;229E月分布较为分散,各月份间差异无统计学意义（x：=15．651,P〉0．05）,在各年龄组的分布呈不集中趋势;NL63流行高峰为6-9月,与其他月份比较差异有统计学意义（x2=5．801,P〈0．05）,检出率最高的年龄组为7～17岁（1．03％）。结论广州市2010—2012年冠状病毒流行高峰为1月份。老年组检出率最高;亚型中OC43阳性率最高,NL63次之,229E亚型阳性率最低;0C43流行高峰为冬季,老年组检出率较高;229E流行高峰及感染人群较为分散;NL63流行高峰为夏季,儿童青少年组检出率最高。     

Taqman荧光定量RT—PCR在呼吸道感染患者临床标本非SARS冠状病毒检测中的应用

目的建立检测四种常见人非SARS冠状病毒核酸特异的快速、敏感的TaqManqRT—PCR检测方法,应用于急性呼吸道感染患儿的感染分析。方法分别应用TaqManqRT—PCR与普通RT—PCR平行检测248份呼吸道标本,对方法的灵敏性、特异性和稳定性以及临床标本的适用性进行比较评价,阳性标本以体外转录RNA为标准品进行病毒载量定量。结果本方法可对HKU1、NL63、229E、OC43四种冠状病毒进行特异性诊断,与其他病毒无交叉反应,检测灵敏度可达10拷贝／μl,检测线性范围可达10^1～10^8拷贝／μl,248份标本中HKU1、NL63、229E、OC43阳性率依次为1．2％,0．8％,1．2％,1．6％,其中OC43荧光RT—PCR法检出率高于普通RT—PCR,其余三种病毒两种方法检测结果一致。非SARS冠状病毒阳性标本均检出于12月至次年5月。结论建立的TaqManRealtimeRT-PCR法具有特异性强、灵敏性高的特点,是开展非SARS冠状病毒的临床检测与疾病监测的有效技术手段。     

人冠状病毒229E、OC43与SARS-CoV血清学相关性的研究

目的 检测正常人和SARS患者血清中3种人冠状病毒（229E、OCA3和SARS-CoV）特异性抗体．分析3种冠状病毒血清学相关性。方法 采用免疫印迹、免疫荧光和ELISA方法检测100例健康献血员、34例SARS患者恢复期以及11例SARS患者双份血清中229E、OCA3和SARS-CoV3种冠状病毒核衣壳（N）蛋白抗体。结果 用免疫荧光方法检测100例健康献血员血清中229E、OCA3和SARS-CoV IgG阳性率分别为98％、100％和1％，34例SARS患者恢复期血清中3种冠状病毒IgG的阳性率均为100％；免疫印迹检测100例健康献血员血清中229E、OCA3和SARS-CoVN蛋白IgG阳性率分别9r7％、99％和2％，34例SARS患者恢复期血清中229E、OCA3和SARS-CoVN蛋白IgG阳性率分别97％、100％和100％；11例SARS患者的急性期和恢复期双份血清中，免疫荧光检测有5例出现229E IgG滴度4倍或以上升高，10例出现OC43 IgG滴度4倍或以上升高，ELISA检测2例出现229EN蛋白IgG滴度4倍以上升高，没有一例出现OCA3N蛋白抗体滴度升高。结论 正常人群中普遍存在229E和OCA3两种人冠状病毒抗体，SARS-CoV感染者存在对人冠状病毒229E和OCA3血清学交叉反应，提示核衣壳蛋白不是引起血清学交叉反应的主要抗原，结果对研究SARS溯源有重要意义。     

深圳市流感样病例感染常见呼吸道病毒的流行特征分析

目的 了解深圳地区急性呼吸道感染患者中15种常见呼吸道病毒及其亚型的感染状况.方法 于2012年10月-2013年9月在深圳市两家流感监测哨点医院采集了1603例流感样病例的咽拭子样本,应用荧光定量PCR方法检测15种常见呼吸道病毒及其亚型,分析病例流行病学特征.结果 1603份标本中共检出呼吸道病毒阳性801份,阳性率为49.97％.鼻病毒的检出率最高,为15.53％(249/1603),其它检出率较高的病毒为甲型流感病毒(12.10％,194/1603)、呼吸道合胞病毒B型(6.8％,109/1603)、腺病毒(5.24％,84/1603)、副流感病毒3型(2.87％,46/1603)、间质肺病毒B型(2.37％,38/1603)、乙型流感(1.37％,22/1603)和冠状病毒OC43型(1.06％,17/1603).呼吸道病毒总阳性率3月份最高,11月份最低.各种病毒的流行季节不同,鼻病毒夏季高发,呼吸道合胞病毒在冬春季和夏季流行,腺病毒在冬季和夏季高发.0-4岁和5-14岁组儿童的呼吸道病毒阳性率(分别为60.4％和49.3％)高于其它年龄组.结论 深圳地区存在多种呼吸道病毒流行,鼻病毒、甲型流感病毒、呼吸道合胞病毒和腺病毒为主要病原体,其流行季节各不相同,低年龄组儿童为呼吸道病毒感染的高危人群.     

2010—2012年韶关市儿童呼吸道感染病毒病原学分析

目的探讨韶关市儿童常见呼吸道感染的病原学特点和分布特征。方法采集2010年7月至2012年7月因呼吸道感染于粤北人民医院的住院患者呼吸道标本171份,采用荧光定量PCR方法,对呼吸道标本同时进行甲型流感病毒（FluA）,乙型流感病毒（FluB）,腺病毒（ADV）,博卡病毒（BoV）,副流感病毒1型（PIV1）、2型（PIV2）、3型（PIV3）,鼻病毒（HRV）,呼吸道合胞病毒（RSV）,冠状病毒229E、0C43、HKUl、NL63,偏肺病毒（MPV）等14种常见呼吸道病毒核酸检测。结果171份标本中检出阳性标本93份,核酸阳性率为54．4％（93／171）,其中FluA占首位,阳性率为8．2％（14／171）,其他依次为ADV7．6％（13／171）,HRV7．6％（13／171）,PIVI／II／III 7．0％（12／171）,RSV 6.4％（11／171）,FluB5．8％（10／171）,BoV5．3％（9／171）,MPV3．5％（6／171）,冠状病毒（HCoV—OC43／HKUl）2．9％（5／171）。不同性别问患儿呼吸道病毒阳性率差异无统计学意义（P〉0．05）。≥6月龄组阳性率最低（37．5％）,1—3岁年龄组阳性率最高（62．1％）。结论韶关地区儿童发热呼吸道病毒感染病例的病原体以甲型流感病毒、腺病毒和鼻病毒为主。     

广东省深圳市流感流行特征分析

目的 掌握深圳市流感流行规律,了解甲流大流行以后流感的流行趋势.方法 对深圳市流感样病例监测数据、病原学检测结果和暴发疫情资料进行分析.结果 深圳市的流感样病例百分比(ILI％)为4.67％,呈逐年下降趋势,ILI年龄构成以0-4岁为主(占54.2％).流感病毒分离平均阳性率为10.6％,按月分析流感病毒分离阳性率与ILI％变化趋势呈正相关(r=0.447,P =0.001).全市报告了482起ILI暴发疫情,以乙型流感为主(占63.9％).2010年深圳市季节性流感出现了春季和夏季流行高峰,分别以乙型Victoria系和甲型H1N1亚型为优势株;2011年为冬春季和秋季高峰,以甲型H1N1和季节性H3亚型为优势株;2012年出现了冬春季和夏季高峰,以乙型(Victoria系和Yamagata系)和季节性H3亚型为优势株;2013年出现了春、秋、冬季三个流行高峰,分别以甲型H1N1、季节性H3和乙型Yamagata系为优势株.结论 深圳市季节性流感每年均出现2-3个流行高峰,分别在冬春季和夏秋季,每年流行高峰出现的时间不同,每年流行的型别不同.     

长沙地区住院儿童冠状病毒HKU1的感染状况及临床特征

目的了解冠状病毒HKU1在长沙地区急性下呼吸道感染住院儿童中的感染状况及临床特征。方法采集2007年9月至2008年8月长沙地区因急性下呼吸道感染住院儿童鼻咽抽吸物（nasopharyngeal aspirates,NPA）样本1165份,用RT-PCR方法检测冠状病毒HKU1 pol基因,测序分析阳性扩增产物,统计分析临床资料。结果1165份NPA样本中,检测出12份冠状病毒HKU1,阳性率为1．03％（12／1165）,患儿年龄最小8天,最大3岁,临床诊断多见支气管肺炎（83．33％）,所测核苷酸序列与冠状病毒HKU1参考株同源性为98．18％～100％。结论冠状病毒HKU1阳性患儿年龄均在3岁及3岁以下;性别无差异;冬春季检出率高;单一基因型在长沙地区流行;HKU1也是我国儿童急性下呼吸道感染中的一种病原。     

2007年秋冬季北京地区成人发热就诊患者中人冠状病毒229E感染状况分析

目的 了解北京地区成人发热就诊患者中人冠状病毒HCoV-229E感染病原学、临床表现和流行病学特点.方法 2007年10月至12月从北京地区发热且有呼吸道症状的就诊成人中采集158份鼻咽拭子标本,利用荧光定量RT-PCR法进行HCoV-229E筛查;利用常规PCR方法扩增HCoV-229E基因片段测序;对感染阳性患者进行HCoV-NL63、HCoV-HKU1及HMPV的共感染筛查、临床表现描述和流行病学特点分析.结果 158份鼻咽拭子标本中荧光定量RT-PCR检测出103(65.2%)例HCoV-229E阳性;其中26例存在与HCoV-NL63(3例)、HCoV-HKU1(3例)及HMPV(20例)的混合感染;HCoV-229E阳性标本的临床表现以头痛(70.9%)、咽红(70.9%)、咽痛(69%)、寒颤(68%)、咳嗽(33%)、有痰(21.3%)、流涕(21.4%)和鼻塞(16.5%)为主,少量可见呕吐(6.8%)、呼吸困难(3.9%)和腹泻(1.9%);统计学分析表明其感染与性别、年龄无关.结论 人冠状病毒HCoV-229E感染在秋冬季成人发热就诊患者中为常见病原,可合并感染其他病毒.其临床表现呈上呼吸道感染特征.2007年秋冬季在北京地区成人中可能存在HCoV-229E局部流行.     

276例手术切除肺腺癌亚型与EGFR/ALK基因状态

目的:探讨手术切除肺腺癌各亚型EGFR和ALK基因状态分布.方法:应用ARMS方法检测手术切除肺腺癌石蜡组织中EGFR基因突变和ALK融合基因情况.结果:276例肺腺癌手术样本中,EGFR基因突变率为54.71％(151/276),其中19del为28.99％(80/276),L858R为23.19％(64/276),20-ins为0.72％(2/276),L861Q为0.72％(2/276),G719X为1.09％(3/276),S768I为0.36％(1/276)和T790M为0.72％(2/276),其中包含G719X+S768I,19del+T790M,L858R+T790M各1例,ALK基因融合阳性率为5.80％(12/207),在肺腺癌各亚型中EGFR基因突变附壁状腺癌,腺泡状腺癌,乳头状腺癌,实体状腺癌和浸润性黏液腺癌之间差异有统计学意义(P＜0.001,P=0.009,P=0.023,P＜0.001和P=0.030),与其他类型之间差异均无统计学意义(P＞0.05);在肺腺癌各亚型中ALK融合基因突变各亚型之间差异均无统计学意义(P＞0.05).结论:肺腺癌组织学亚型与EGFR基因突变有关,附壁状腺癌、腺泡状腺癌和乳头状腺癌出现EGFR基因突变比其他亚型更加明显.     

兰州地区急性呼吸道感染患儿冠状病毒NL63的检测与临床研究

目的 探讨兰州地区冠状病毒NL63(HCoV-NL63)在急性呼吸道感染患儿中的流行现状及临床特点.方法 收集2006年11月至2009年10月兰州大学第一附属医院急性呼吸道感染(ARTIs)患儿1169例鼻咽分泌物,应用RT-PCR方法检测HCoV-NL63以及其余7种常见呼吸道病毒:鼻病毒(HRV),呼吸道合胞病毒(RSV),偏肺病毒(hMPV),流感病毒(IFVA,IFVB)副流感病毒1-3(HPIV1-3)及PCR方法检测腺病毒(ADV),博卡病毒(HBoV).结果 检测出HCoV-NL63阳性标本35例,检出率2.99％,2007年8、9月,2009年7、8月检测阳性标本阳性率较高,分别为23.53％、17.65％,50％、33.33％.2007年12月至2009年2月未检出HCoV-NL63阳性标本.25(25/35)例混合其他病毒感染,混合感染率为71.43％,最常见的混合感染病毒是HRV.3岁及以下和3岁以上HCoV-NL63感染组感染率差异无统计学意义.HCoV-NL63阳性患儿主要的诊断是支气管肺炎和毛细支气管炎,主要的症状是发热和咳嗽.HCoV-NL63单独感染组和混合感染组除消化道症状外,在其余症状和临床诊断方面,差异均无统计学意义.结论 HCoV-NL63是兰州地区呼吸道感染患儿的重要病原,夏季是兰州地区HCoV-NL63感染高峰期,HCoV-NL63的流行存在年度差异.HCoV-NL63感染存在很高的混合感染率,混合感染并不加重HCoV-NL63感染的病情.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

The universal muscular chain reaction,muscle spasm,Torsions, ruptures and extravasations. Chameleons of pathology and some manifestations of simple muscular disorders

Most bodily functions require the coordinated actions of complementary and supplementary paired muscle groups. Where this essential muscular cooperation is lacking, hollow organs may burst and others become literally screwed up, giving rise to many similar spastic diseases such as Torticollis, Twisted ovarian cyst, Torsion of the Testis, Volvulus of the intestines, Varicose Veins, Megacolon, Aortamegaly, Scoliosis, Erb's Palsy, Peyronie's Disease, Main-en-Griffe, Undescended Foot (Pes Cavus), Talipes, Strabismus. Spasm is “panenepidemic” and unclassified examples of Torsion Dystonia and Dyskinesia really are as common as debt and taxes.     

Class II HLA in Georgia Caucasus Tbilisi Georgians and their Mediterranean ancestry: The Usko Mediterranean languages

Georgia (or Sakartvelo in its own language) is a South Caucasus Mts. country with its easternmost part is enigmatically named Iberia, like the Iberian Peninsula, which may refer to rivers “Kura” and “Ebro” or their valleys respectively. Most of their inhabitants speak Georgian which is included within Dene-Caucasian group and Usko-Mediterranean subgroup of languages. The latter includes Basque, Berber, ancient Iberian-Tartessian, Etruscan, Hittite, Minoan Lineal A and others. In the present paper, HLA class II -DRB1 and -DQB1 alleles has been studied and extended haplotypes calculated. Most frequent haplotypes are also of Mediterranean origin (i. e.: (A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*51)-DRB1*13:01-DQB1*06:03, or (A*24-B*35)-DRB1*01:01-DQB1*05:01) and DA genetic distances show that closest world populations to Georgians are Mediterraneans. Georgians also show common extended haplotypes ((A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*13)-DRB1*07:01-DQB1*02:01 and (A*03-B*35)-DRB1*11:01-DQB1*03:01) with Svan people, a secluded population in North Georgia mountains. We can conclude that Georgians belong to a very old Mediterranean substratum according to both linguistics (Usko Mediterranean languages) and HLA genetics.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.