我国HIV-1主要流行株外膜蛋白(env)基因V3～V4区变异及其与生物学特性的关系

目的　研究我国HIV 1主要流行毒株亚型的envV3～V4区变异与生物学特性的关系。方法　应用nested PCR对 1 57份获自我国 1 2个省份的HIV 1毒株env区序列进行扩增 ,并使用ABI 377型测序仪测序 ,然后应用BLAST、GCG和MEGA等生物学软件或程序对env基因V3～V4区序列进行分析。结果　B′亚型毒株V3顶端四肽存在着 4种类型 :GPGR ( 54% )、GPGQ ( 2 8% )、GPGK( 1 6 % )和GPGA( 2 % ) ,B′/C重组毒株全部为GPGQ( 1 0 0 % ) ,CRF0 1 AE重组毒株呈现GPGQ( 95% )和GPGR( 5% )两种类型 ;B′/C和CRF0 1 AE重组毒株V3～V4区及其临近区域N 糖基化位点比B′亚型毒株N 糖基化位点保守。而B′亚型毒株V3环的净电荷分别显著高于B′/C和CRF0 1 AE毒株 (P <0 .0 1 ) ;根据V3环关键氨基酸推测辅助受体使用情况的结果显示 :B′亚型毒株有 9.2 6 %可能使用CCR5,7.4 1 %可能使用CXCR4 ,其余 83.33%不能对辅助受体的使用作出预测。所有B′/C重组毒株被预测可能使用CCR5。CRF0 1 AE重组毒株有 90 .4 8%被预测可能使用CCR5,没有被预测为使用CXCR4的序列 ,9.52 %不能作出预测。结论　B′亚型毒株大部分可能为NSI型 ,少部分可能为SI型 ,而B′/C和CRF0 1 AE重组毒株绝大部分为NSI型。我国主要流行株的V3～V4区尤其是V3环的氨基酸     

中国HIV-1型B、C亚型主要流行株外膜蛋白基因V3-V4区序列的变异分析

目的　研究我国人类免疫缺陷病毒 1型 (HIV 1)B、C亚型主要流行株在感染过程中基因变异的特点及其与选择压力的关系。方法　应用巢式聚合酶链反应 (nested PCR)对 2 5 8例HIV 1感染者血样中的HIV 1外膜蛋白 (env)基因进行扩增 ,并使用ABI 377型测序仪对扩增产物测序后 ,选择其中 37份B亚型和 35份C亚型HIV 1毒株env基因包括V3～V4区的序列进行比较分析 ,并计算和分析氨基酸同义替换与非同义替换的比值 (Ks Ka)。结果　B亚型毒株V3～V4区的基因离散率高于C亚型毒株。无论B亚型 ,还是C亚型毒株 ,其V4区基因序列较V3区变异更大。在C亚型毒株中 ,V3区基因序列变异甚至比V3上游区和C3区小。B和C亚型毒株整个V3～V4基因区的Ks Ka比值均 <1,差异有非常显著性 (P <0 0 0 1) ,其中B亚型毒株以V3区的Ks Ka比值最小 ,而C亚型毒株则以V4区的Ks Ka比值最小。结论　B和C亚型毒株env基因的变异主要发生在V4区而不是V3区。C亚型毒株V3区较V3上游区和C3区还要保守 ,是本研究的特殊发现。这两种亚型在我国快速流行中发生的变异是在选择压力下发生的 ,而不是随机进化的结果 ,而且选择压力对这两种亚型毒株V3、V4区的作用程度也不一样。这将为我国艾滋病防治策略的制定和疫苗研究提供科学的依据。     

1990～1997年云南IDUs HIV-1 B亚型分离株膜蛋白V3环顶端四肽基因序列的变化趋势

目的　观察云南静脉药瘾 (IDUs)HIV 1分离株env基因V3环顶端四肽氨基酸和相应核苷酸序列随时间推移的变化。方法　根据 1990～ 1997年间 6 2株HIV 1分离株env基因C2 V3区DNA序列 ,对HIV 1膜蛋白V3环顶端四肽基因序列 (基序 )及其编码核苷酸进行分析 ,并探讨其随时间推移的变化趋势。结果　 1990～ 1997年间 6 2株HIV 1毒株膜蛋白V3环顶端四肽基序有程度不同的氨基酸变异 ,主要表现在第四位氨基酸的改变 ,变异呈现一定的趋向性。 6 2株云南标本中有 46例(74 2 % )为GPGR ,10例 (16 1% )为GPGQ ,4例 (6 4% )为GPGK ,另有 2例 (3 3% )为GPER且四肽基序编码核苷酸的变异主要表现为A→G的改变。结论　 1990～ 1997年这一时期内 ,云南IDUs特定感染者范围内HIV 1分离株膜蛋白V3环顶端四肽序列已表现出较大的趋向性 ,符合HIV 1国际B亚型SF2株的GPGR模式     

HIV1膜蛋白PND编码基因自然变异株的发现

目的 对1例来自华东地区HIV1分离株（WWBH7）的前病毒env基因C2-V3区进行序列分析。方法 以HIV1感染者外周血单个核细胞基因组为模板进行套式PCR扩增HIV1env基因C2-V3区片段,将此扩增产物插入T-Vector,酶切鉴定重组质粒,使用AB1737自动DNA序列测定仪测定序列并用DNASIS软件进行分析。结果 DNA序列资料显示该毒株属于HIV1B亚型衍生株,但与HIV1B亚型的标准株如SF2株相比,该HIV1毒株的env基因V3区下游有192bp的重复插入突变,使得该毒株的膜蛋白PND编码基因呈现双V3区的变异该段DNA序列已登录于GenBank(AF220245)。结论 该分离株是1个在膜蛋白PND编码基因有大片段插入突变的HIV1变异株。     

福建省HIV-l流行毒株的基因序列测定和亚型分析

目的　通过DNA序列分析 ,确定福建省HIV 1流行毒株的亚型。方法　从福建省HIV 1抗体阳性者中选择 9例经性接触感染HIV 1者 ,采集其全血分离周围血淋巴细胞 (PBMCs)提取DNA作为PCR扩增的模板 ,设计合成 3对套式引物扩增HIV 1前病毒DNA的C2～V3区用于测序。所测序列与HIV 1中的 6个亚型的 10个国际标准或参考序列进行比较 ,确定被检标本的型别。结果使用PCR技术对 9份福建HIV 1阳性感染者PBMC样品进行扩增 ,获得HIV 1膜蛋白 (env)基因的核酸片段 ,并对其约 2 47bp核苷酸序列进行了分析、比较 ,证实 9份样品中 7份为E亚型 ,其余 2份分别为B和C亚型毒株。E亚型各株间序列存在一定差异 ,组内基因离散率为 6 2 8± 2 40。结论　福建省目前HIV 1的流行株主要为E亚型 ,流行株的来源较为复杂 ,株间序列差异不仅与感染时间有关 ,也和感染地点不同等因素有关 ,这些特点不同于我国其他一些省份发现的经血传播的HIV 1B或C亚型 ,其流行时间不长 ,株间序列差异不大     

广西HIV-1 CRF01-AE重组毒株env基因V3环序列变异及其与生物表型的关系

目的研究广西HIV-1 CRF01-AE重组毒株env基因V3环序列变异及其与生物表型间的关系。方法从广西主要流行区收集来的50份HIV-1感染者血液样本中提取前病毒DNA，使用巢式聚合酶链反应（nested-PCR）扩增HIV-1 env基因片段并进行亚型鉴定，选择38份CRF01-AE重组型HIV-1毒株env，基因V3环及邻近区域的序列进行系统树和氨基酸变异分析。结果38份CBF01-AE重组毒株中36份与分离于广西地区的CRF01-AE.97CNGX2f和泰国代表毒株THCM240接近，另外2份与中非共和国代表株90CF402聚成一簇；CRF01-AE重组毒株V3环顶端四肽存在着4种类型：CPCQ、GPGR、GPGH和GPGA；根据V3环关键氨基酸推测辅助受体使用情况，结果显示：71．05％的CRF01-AE重组毒株可能使用CCR5作为辅助受体，28．95％不能对其辅助受体的使用情况做出预测。结论广西HIV-1 CRF01-AE重组毒株V3顶端四肽变异较大，而且大部分毒株可能为NSI型。这可为广西该毒株的防治和诊断试剂的更新提供参考。     

河北省HIV-1流行株的env基因序列测定和亚型分析

目的了解河北省HIV-1亚型的分布特点和传播方式,推测流行时间,预测流行趋势。方法采集HIV感染者的全血样品,分离外周血单核细胞(PBMC),提取前病毒DNA,使用套式聚合酶链反应(nested-PCR),扩增HIV-1的env基因的C2-V3区并进行序列测定和亚型分析。结果对22份HIV-1感染者的样品,扩增得到了18份HIV-1envC2-V3基因片段,经序列测定和基因分析鉴定出3种HIV-1M亚群基因亚型,即:B′、CRF-BC和C亚型。B′亚型的组内基因离散率为7.84%±3.14%(n=14),基因序列与云南瑞丽株rl42(泰国B亚型)相近;2株CRF-BC亚型与广西毒株的基因离散率为4.60%。与血液途径感染有关的人员均为B′(泰国B)亚型。结论目前,在河北省发现了3种HIV-1亚型。输供血途径中B′亚型仍是主要的流行亚型,可能来源于云南吸毒人群。HIV-1B′亚型在河北省的流行时间大约为7~9年。     

北京市男男性接触HIV-1感染者膜蛋白基因序列测定及亚型分析

目的 了解北京市男男性接触人群(MSM)中HIV-1的最新流行趋势及膜蛋白V3环序列特征.方法 巢式聚合酶链式反应(n-PER)扩增2007年提取的北京市男男性接触HIV感染者基因组DNA样品,对膜蛋白基因C2-V3区测序,进行病毒亚型及V3环序列特点分析.结果 11例样本中,4例是欧美B亚型,5例是AE重组亚型,1例是BC重组亚型,1例是01B重组亚型.V3环顶端四肽以GPGQ和GPGR为主.结论 北京市男男性接触HIV-1感染者中重组亚型呈蔓延流行趋势.     

中国东北地区未经抗病毒治疗的HIV/AIDS患者HIV毒株的耐药基因变异研究

目的　研究我国东北地区未接受抗逆转录病毒治疗的HIV AIDS患者HIV毒株的逆转录酶和蛋白酶耐药变异情况 ,为开展大规模临床抗病毒药物治疗提供本底数据。方法　RT PCR和套式PCR扩增HIVpol区基因 ,双脱氧法测定逆转录酶和蛋白酶基因序列 ,与国际耐药数据库比对辨别耐药变异。结果　 (1) 5 3例患者毒株亚型分析结果 :B′亚型 4 7例 ,B′ C亚型 4例 ,A、B亚型各 1例 ;(2 )未发现逆转录酶和蛋白酶原发耐药变异存在 ,但发现存在逆转录酶抑制剂继发变异 :M4 1L(1.9% )、I6 3M (1.9% )、L74I (1.9% )、S6 8G (1.9% )、V75L (3.8% )、V10 6I (1.9% )、I135L T (5 .7% )、V179D (7.5 % )和V189I (1.9% ) ,无症状感染者RT继发耐药变异出现率为 11.8% ,而艾滋病患者为5 2 .6 % (P <0 .0 1)。存在大量蛋白酶耐药继发变异V77I (88.7% )、L6 3P (86 .8% )、E35D (81% )、A71V(2 4 .5 % )、R4 1K (15 .1% )、L10I (9.4 % )、R5 7K (9.4 % )、D6 0E (9.4 % )、N37D (5 .7% )、G16E (3.8% )、I15V (1.9% )、M36I (1.9% )、K5 5R (1.9% )和L89M (1.9% )。未发现明显的亚型特异性耐药变异。结论　在中国东北地区未接受抗逆转录病毒治疗的HIV AIDS患者中未发现毒株逆转录酶和蛋白酶耐药原发变异 ,但大量继发耐药变异的存在提     

32例有偿献血感染者HIV-1膜区序列测定及V3环氨基酸变异特点分析

目的 了解我国有偿献血人群中HIV-1B'亚型流行株的膜蛋白V3环序列特征.方法 巢式PCR扩增前病毒DNAc2-c3区后测序,进行病毒亚型鉴定和V3环序列特点分析.结果 所检的32个HIV-1B'亚型株V3环顶端四肽有五种形式.AIDS组与无症状感染组相比前者呈现更多的T/SI型毒株V3环变异特点.结论 我国有偿献血人群中HIV-1 B'亚型流行株V3环顶端四肽有多种形式.处于病程不同时期的患者体内病毒V3区呈现不同的氨基酸变异特点和电荷积累趋势,提示可能具有不同的生物学表型特征.     

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.