不孕患者中宫颈HPV高危型别感染率及病毒载量的调查

目的调查高危型人乳头瘤病毒（HPV）在不孕患者宫颈细胞中的感染情况,探讨HPV病毒型别及其载量（viral load）对不孕发生的影响。方法对临床130份不孕患者的宫颈脱落细胞标本和在门诊体检的150份对照组的宫颈脱落细胞标本采用多通道实时荧光定量PCR仪进行八种高危HPvDNA分型及定量检测,该八种高危HPV型别为主要高危型：HPV16,18,45,31和次要高危型HPv33,52,58,67。结果不孕组阳性率为25．38％（33／130）,对照组的阳性率为11．33％（17／150）,两组间的阳性率差异有统计学意义。不孕组的33份阳性标本中病毒载量≥10^6为24例,病毒载量〈10^6为9例;对照组的17份阳性标本中,病毒载量≥10^6为4份,病毒载量〈10^6为13份,两组间的病毒载量有显著性差异。结论不孕组高危型HPV感染率比正常人群（对照组）高。对不孕组病毒载量的分析表明：不孕组人群中其病毒载量明显高于正常人群。此外,本研究还为分泌物核酸检测的定量设置了内标（β-球蛋白）,提出可供临床使用的分泌物取样的核酸定量检测方法。     

先天性HCMV感染新生儿尿液中病毒载量与疾病严重度的关系探讨

目的探讨新生儿尿液中人巨细胞病毒（HCMV）病毒载量与先天性感染的关系及与HCMV所致疾病严重程度的关系。方法采集98例经PCR方法确诊的有症状及无症状的先天性HCMV感染新生儿尿液标本,用实时荧光定量PCR法（FQ—PCR）检测尿液中巨细胞病毒载量。结果98例先天性HCMV感染的新生儿中85例在出生后有临床症状（86％）。无症状感染和有症状感染的新生儿尿液中平均HCMV病毒载量分别是1．4×10^5拷贝／ml和3．1×10^6拷贝／ml,P〈0．01,差异有统计学意义。结论先天性HCMV感染的新生儿中无症状感染者尿液中病毒载量显著低于有症状感染者,提示病毒载量与疾病严重程度相关。     

基因芯片分型与实时荧光PCR筛查成都地区妇女宫颈HPV-DNA感染结果比较

目的了解成都地区HPV亚型的感染情况;评价基因分型（genotyping）与实时荧光定量PCR（real-time PCR）两种人乳头瘤病毒（HPV）检测方法的差异性,为临床应用提供参考。方法对2012年1月-2013年6月间来院就诊的女性,其中3537名进行基因芯片检测,991名进行实时荧光PCR检测。所得数据用SPSS 17.0进行统计分析。结果基因芯片分型法检测出阳性1315例,阳性率37.18%;实时荧光PCR检测出阳性137例,阳性率13.82%,两种方法检出率差异明显。基因分型法显示,HPV6、11、16、58、43型位居前五;荧光定量PCR显示HPV6、11、16、52、58在前五位。结论成都地区低危感染以HPV 6、11型为主,高危以16型为主,次之52、58型;基因芯片分型与临床HPV-DNA筛查检出率高,优于荧光定量PCR法。     

荧光定量聚合酶链反应检测人乳头瘤病毒6/11型

目的 了解女性阴道炎患者人乳头瘤病毒6／11型（HPV6／11）的感染情况。方法 用荧光定量聚合酶链反应（FQ－PCR）技术检测129例门诊患者阴道分泌物中HPV6／11的病毒拷贝数。结果 共检出HPV6／11阳笥者47例,阳性率为36．43％;拷贝数在10^5以上者39例,占阳性者中的82．98％;40岁以上年龄组阳性率高且病毒复制量均在10^5以上。结论 中老年阴道炎患者就诊晚,感染重。FQ－PCR检测HPV敏感、快速、准确,特别是其定量特点对临床很有意义。     

荧光定量PCR法在重组逆转录病毒pLXSN-BDNF滴度测定中的应用

目的制备含大鼠脑源性神经营养因子（BDNF）基因的逆转录病毒，建立荧光定量PCR测定滴度的方法。方法扩增BDNF基因，构建重组质粒pLXSN-BDNF，将其转染包装细胞，经G418筛选、病毒浓缩后用荧光定量PCR法和克隆形成法测定滴度。结果经PCR、酶切和测序鉴定，BDNF基因成功插入逆转录病毒载体中，筛选得到稳定的分泌重组病毒细胞系。重组病毒100倍浓缩后经荧光定量PCR法和克隆形成法测定的滴度分别为6．92×10^6拷贝/ml和3．2×10^5CFU/ml，两种方法测得的结果差异有统计学意义（P〈0．05）。结论成功扩增BDNF基因，制备了重组病毒pLX—SN-BDNF，建立了荧光定量PCR检测滴度的方法，为基因治疗青光眼性视神经损伤的实验研究提供重要参考指标。     

HIV合并HPV阳性标本中HPV的型别检测

目的探讨深圳地区人类免疫缺陷病毒（HIV）阳性人群中人乳头瘤病毒（HPV）感染分型情况,为HIV阳性人群中HPV感染的防治提供依据。方法运用荧光PCR方法和反向斑点杂交技术对HPV阳性患者进行HPV分型检测。结果在HIV感染者中。利用反向点杂交分型方法对40例HPV阳性标本进行HPV基因分型。其中单型感染有18例（45．0％）,混合感染有19例（47．5％）,共检出16种HPV型别,其中包括11种高危型（16、18、31、33、35、45、52、56、58、68、73型）和5种低危型（6、11、42、44、54型）。在16种型别中,感染率最高的为16型（25．0％）,其次为52型（17．5％）、58型（15．0％）。结论感染HIV人群中生殖器部位HPV的感染率较高,在HIV阳性人群中检出性病相关的高危HPV16／52／58亚型．对HIV阳性人群中HPV感染的防治有重要指导意义。     

实时荧光定量RT-PCR法定量检测手足口病病原体的探讨

目的了解北京市门头沟区手足口病患儿感染病毒的型别,探索用荧光定量RT—PCR法对手足口病患者咽拭子标本中肠道病毒71型（EV71）、柯萨奇病毒A16（CoxA16）型病毒载量进行定量检测的可行性。方法采用实时荧光RT—PCR体外扩增法对81例手足口病患儿咽拭子标本提取的RNA进行检测。结果28例手足口病患者体内CoxA16病毒载量〉10^3 copies·ml-1。实时荧光定量RT—PCR法构建的标准曲线显示,样本阈值环数（Ct）值与病毒拷贝数的对数（10g10）之间的相关系数为0．9998,相关性良好。4例手足口病患者咽拭子标本中EV71型病毒载量〉10^3 copies·ml-1。实时荧光定量RT-PCR法标准曲线显示,Ct值与病毒拷贝数的对数之间的相关系数为0．9996,相关性较好。结论门头沟区手足口病病原谱以CoxA16型为主,34．6％的手足口病患儿CoxA16型病毒载量〉10^3 copies·ml-1,4．9％的患儿EV71型〉10^3 copies·ml-1。实时荧光定量RT-PCR法对咽拭子标本中CoxA16、EV71核酸定量检测比较简便快速、结果直观、稳定性强,为进一步探讨患者体内病毒载量与临床症状的关系打下了良好的基础。     

厦门地区女性人乳头瘤病毒感染状况调查

目的 对厦门地区妇女宫颈人类乳头瘤病毒（human papilloma virus,HPV）亚型进行筛查,以探讨其分布规律.方法 采用凯普医用核酸分子快速杂交仪,对7683名2013年1月-2013年12月到厦门市174医院妇科门诊或病房就诊的女性进行生殖道21种HPV感染基因亚型筛查.结果 7683例样本中,HPV感染者1421例,阳性率18.50％.感染人群主要集中在30～39岁.其中高危型HPV阳性率为16.01％,低危型HPV阳性率为2.49％.单一基因型别者1148例,阳性率为14.94％;双重感染者220例阳性率为2.86％;三重以上感染者53例,阳性率为0.70％;HPV感染阳性率居前6位的基因型分别为HPV-52（3.64％）、HPV-16 （3.33％）、HPV-58（2.98％）、HPV-53（2.16％）、HPV-CP8304（1.91％）和HPV-18（1.09％）.结论 厦门地区妇女HPV感染率高,且以高危型单一基因亚型感染为主;感染基因型别主要以HPV-52、16、58、53、CP8304和18为主,具有一定的地域差异性;52及58型感染率高,对于疫苗的研制和开发有参考意义.     

北京市双桥医院妇科门诊患者人乳头瘤病毒感染情况调查

目的了解北京市朝阳区双桥医院妇科门诊不同年龄女性患者人乳头瘤病毒（human papilloma virus,HPV）亚型感染情况。方法采用核酸分子快速导流杂交技术,对2009—2012年835例受检者宫颈分泌物进行HPV—DNA亚型检测。结果538例女性患者中共检测到175例HPV阳性病例,其中高危亚型170例,低危亚型5例。高危亚型检出率居前5位依次为HPV-16（31．4％）、52（20．0％）、58（17．1％）、56（14．3％）、66（14．3％）。≥40岁以上年龄组HPV阳性率显著高于40岁以下年龄组。结论高危型HPV-16、52、58、56、66亚型是北京市双桥医院人乳头瘤病毒感染的主要亚型,40岁以上人群是防治的重点。     

机会性筛查人群中高危型人乳头瘤病毒与宫颈病变相关性的分析

目的研究第二代杂交捕获法（HC2）高危型人乳头瘤病毒（HR—HPV）检测阳性样品中HPV与宫颈病变的相关性。方法随机抽取HC2阳性患者602名（344名行细胞学检查）,采用PCR一反向点杂交（RDB）法对HPV分型并分析HPV型别与细胞学的相关性。结果602例患者PCR阳性率94．5％;HPV分型检出率依次为HPVl6、52、58、56、39等;HC2检测谱外单独感染24例;HPVl6、HPVl6／18与ASCUS、LSIL、HSIL有明显相关性。结论机会性筛查人群中最常见HPV型别依次为HPVl6、52、58;HC2检测的假阳性率为4．0％;HPVl6与宫颈病变程度相关。     

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.     

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.     

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.     

'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.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

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.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.