中国人WD基因第14和18号外显子的错义突变

目的了解中国人肝豆状核变性（Ｗｉｌｓｏｎ＇ｓｄｉｓｅａｓｅ,ＷＤ）基因第１４和１８号外显子的突变情况,与国外报道的这两个外显子的高突变频率进行比较。方法应用聚合酶链反应－单链构像多态（ＰＣＲ－ＳＳＣＰ）结合ＤＮＡ测序技术,对４４例无亲缘关系的ＷＤ患者及６０例正常对照进行突变检测。结果一例患者在１４号外显子发生了Ａｒｇ１０４１Ｐｒｏ（３１２１Ｇ→Ｃ）纯合错义突变,另一例患者在１８号外显子发生了Ａｓｎ１２７０Ｓｅｒ（３８０９Ａ→Ｇ）杂合错义突变。结论Ａｒｇ１０４１Ｐｒｏ为未报道过的新型错义突变,Ａｓｎ１２７０Ｓｅｒ突变与国外报道一致。但均未呈热区分布。     

中国人血友病甲患者FⅧ基因突变研究

为研究血友病甲发病的分子机理，应用聚合酶链反应（ＰＣＲ）结合限制性内切酶ＴａｑⅠ酶切分析和ＰＣＲ结合变性梯度凝胶电泳（ＰＣＲ－ＤＧＧＥ），分别研究了７４名中国血友病甲患者ＦⅧ基因外显子１８、２２～２４、２６和外显子８与１４的３′端的基因突变情况，结果检测到一例基因突变，该突变位于第２４号外显子２２０９位密码子，ＣＧＡ－ＴＧＡ，导致了终止密码子的产生。ＰＣＲ－ＤＧＧＥ发现２例基因突变，分别位于外显子８的３４９位密码子，ＧＡＴ－ＧＡＧ，导致Ａｓｐ３４９Ｇｌｕ和外显子１４的１６８９位密码子，ＣＧＣ－ＴＧＣ，使Ａｒｇ１６８９Ｃｙｓ。血友病甲基因突变的研究将有助于开展遗传咨询和产前诊断工作。     

PCR直接测序在Wilson病基因第8外显子检出一个突变热点

目的寻找中国人Ｗｉｌｓｏｎ病（ＷＤ）基因突变热点。方法应用ＰＣＲ直接测序法对３０例Ｗｉｌｓｏｎ病患者ＷＤ基因第８外显子进行了突变筛查。结果在１４例患者中发现同一种错义突变Ａｒｇ７７８Ｌｅｕ，其中４例为这一突变的纯合，其余为杂合，突变率为３０％。结论ＷＤ基因第８外显子７７８位密码子（ＣＧＧ→ＣＴＧ）系中国人的突变热点之一。     

Wilson病家系ATP7B基因第18外显子突变及多态

目的：检测中国人Ｗｉｌｓｏｎ病（ＷＤ）基因的第１８外显子（ｅｘｏｎ１８）突变及多态。方法：应用多聚酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）技术，分析１６个ＷＤ家系５４例个体和１８例正常人ＡＴＰ７Ｂ基因ｅｘｏｎ１８分子结构改变。结果：在ＷＤ家系中发现有３种单链构象，其中１种为正常构象，１种为突变，另１种可能为正常ＤＮＡ多态；１７例ＷＤ患者及其３７例一级亲属中分别检出突变者１０例和１１例，突变率分别为２９．４％和１４．９％。其中一级亲属检出的突变者中有３例经血清铜氧化酶及眼科Ｋ－Ｆ环检查证实为ＷＤ症状前患者。结论：ｅｘｏｎ１８是中国人ＷＤ基因突变热点之一，大多数ＷＤ患者为复合杂合子；应用ＰＣＲ－ＳＳＣＰ技术分析ｅｘ－ｏｎ１８是一种有效的ＷＤ基因筛选方法，也可为无家族史的ＷＤ可疑者提供诊疗依据     

Wilson's病8号外显子突变研究

目的对中国人ＷＤ基因８号外显子进行突变分析。方法对中国人Ｗｉｌｓｏｎ＇ｓ病（Ｗｉｌｓｏｎ＇ｓｄｉｓｅａｓｅ,ＷＤ）４５例患者以及２０例正常人的ＡＴＰ７Ｂ基因８号外显子进行ＳＳＣＰ分析,对有异常者进行测序,根据突变点序列设计合适的内切酶对所有患者进行酶切分析。结果正常组未见异常。患者组发现ｅｘ－ｏｎ８有泳动异常,序列分析证实Ｇ２２７３Ｔ置换,即Ａｒｇ７７８Ｌｅｕ突变。用限制性内切酶ＭｓｐⅠ对４５例患者以及２０例正常对照进行该位点酶切分析,表明正常组未见异常,患者组有２例突变纯合子,占患者总数４．４％,１１例杂合子,占１２．２％。外显子８的Ａｒｇ７７８Ｌｅｕ突变率占ＷＤ突变基因的１６．６７％。检测了２个突变家系。结论８号外显子突变可能是中国人ＷＤ发病的较重要原因。     

苯丙酮尿症突变基因分析和产前诊断

应用多重等位基因特异ＰＣＲ方法检测分析了３０个苯丙酮尿症家系的５种苯丙氨酸羟化酶基因点突变：外显子７（Ａｒｇ２４３Ｇｌｎ），外显子１２（Ａｒｇ４１３Ｐｒｏ），外显子３（Ａｒｇ１１１Ｔｅｒｍ），外显子１１（Ｔｙｒ３５６Ｔｅｒｍ）和外显子６（Ｔｙｒ２０４Ｃｙｓ）。结果表明，这五种突变占ＰＫＵ基因的４６．６％，其中最常见的突变为前两种，分别占２３．３％和１０．０％。完成了１例ＰＫＵ风险胎儿的产前诊断。     

应用微卫星DNA单体型检测Wilson病症状前患者及杂合子

目的建立快速、准确、有效的Ｗｉｌｓｏｎ病（Ｗｉｌｓｏｎｄｉｓｅａｓｅ，ＷＤ）患者早期诊断及杂合子检测的基因诊断技术。方法应用Ｄ１３Ｓ３０１、Ｄ１３Ｓ３１６、Ｄ１３Ｓ２９６、ＡＦＭ２３８ｖｃ３、ＡＦＭ０８４ｘｃ５等５个微卫星ＤＮＡ（ｓｈｏｒｔｔａｎｄｅｍｒｅｐｅａｔ，ＳＴＲ），经聚合酶链反应（ＰＣＲ），扩增片段长度多态性－聚丙烯酰胺凝胶电泳（ＡＦＬＰ－ＰＡＧＥ），对２３个ＷＤ家系１２０名成员的ＤＮＡ进行单体型分析。结果２例拟诊患者得到确诊。在３１例ＷＤ同胞中，检出肯定症状前患者４例、杂合子８例、正常人１７例、可疑患者２例。结论ＳＴＲ单体型可提供高信息量的遗传诊断信息，为ＷＤ基因诊断提供了重要的新途径     

中国人中所见黑蒙性痴呆的新突变

在３个中国婴儿型黑蒙性痴呆（Ｔａｙ－Ｓａｃｈｓｄｉｓｅａｓｅ，ＴＳＤ）家庭中鉴定了３种突变等位基因，这３家无血缘关系，也无亲近婚配。其中两种突变等位基因是新的，第三种则为曾报道过的突变（Ｇ１４４４Ａ）。家庭１，ＤＮＡ经ＰＣＲ体外扩增后直接测序，发现为ＨＥＸＡ基因第５外显子的第５４７核苷酸处插入一个腺嘌呤所造成的移码突变，导致插入部位下游６ｂＰ处提前出现终止密码。用等位基因特异性寡核苷酸（ＡＳＯ）杂交证明先证者为突变基因纯合子。家庭２，用单链构象多态性（ＳＳＣＰ）分析及将扩增的第１３外显子直接测序，发现为第１４５３核苷酸Ｔ转换为Ｃ，造成第４８５位色氨酸为精氨酸取代，以Ｔ１４５３Ｃ（ｅｘｃｏｎ１３）／Ｗ４８５Ｒ表示之（Ｗ代表色氨酸，Ｒ代表精氨酸）。此突变造成了一个Ｆｎｕ４Ｈ１酶切位点。患儿则为此等位基因的纯合子。当Ｔ１４５３Ｃ定向致突变的αｃＤＮＡ在ＣＯＳ－１细胞中表达时，未能测出氨基已糖苷酶（Ｈｅｘ）Ｓ的活性。家庭３的突变为第１３外显子的１４４４核苷酸Ｇ转换为Ａ，造成第４８２位谷氨酸为赖氨酸取代，以Ｇ１４４４Ａ（ｅｘｏｎｌ３）／Ｅ４８２Ｋ表示之（Ｅ代表谷氨酸，Ｋ代表赖氨酸）。此突变发生于一个ＧｐＧ二核苷酸，以     

es respectively. Conclusion : According to

为了研究中国人Ｗｉｌｓｏｎ病基因（ＡＴＰ７Ｂ基因）第９、１４外显子突变特征。方法：应用聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）银染技术初步研究了１４１便Ｗｉｌｓｏｎ病患者ＡＴＰ７Ｂ基因第９及１４外显子的分子结构改变。结果：通过用ＰＣＲ－ＳＳＣＰ对患者和正常人的ＤＮＡ的电泳带迁移作对比分析，发现在患者中第９和１４外显子ＰＣＲ扩增产物迁移异常者分别为６例（２．１％，６／２８２）和４２例（１４．９％，４２／２８２）。结论：依据ＤＮＡ单链构象与分子电泳迁移的关系，这一初步结果提示该组病人中ＡＴＰ７日基因的第１４外显子可能存在着较高的突变率。     

应用PCR—SSCP银染技术检测Wilson病基因第9,14外显子突变

为了研究中国人Ｗｉｌｓｏｎ病基因（ＡＴＰ７Ｂ基因）第９，１４外显子突变特性，方法：应用聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）银染技术初步研究了１４１例Ｗｉｌｓｏｎ病患者ＡＴＰ７Ｂ基因第９及１４外显子分子结构改变，结果：通过用ＰＣＲ－ＳＳＣＰ对患者和正常人的ＤＮＡ的电泳带迁移作对比分析，发现在患者中第９和１４外显子ＰＣＲ扩增产物迁移异常者分别为６例（２．１％，６／２８２）和４２例（１４．９     

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.     

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.     

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.     

'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.     

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.     

The case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.