β—球蛋白生成障碍性贫血基因分析

本文分析了宜昌地区3个家系8例β—珠蛋白基因突变类型,发现以IVS—2—654(C→T)占多数6/8,次为41—42(-4bp)2/8。3例先证者分别为41—42(-46p)/IVS—2—654(C→T)双重杂合子;IVS—2—654(C→T)纯合子;IVS—2—654(C→T)杂合子。 β—珠蛋白生成障碍性贫血治疗困难,脾切除对于双重杂合子及纯合子患者疗效差于杂合子患者,目前进行产前诊断能达优生目的。     

一个少见的β地中海贫血突变家系

目的　鉴定中国人中 1个少见的 β地中海贫血 (β-地贫 )家系成员的基因转录突变 (- 90C→ T)。　方法　表型分析采用常规的红细胞指数和血红蛋白电泳分析技术 ;反向点杂交技术用于检测中国人 18种已知类型的β-地贫突变 ;β珠蛋白基因全长 DNA测序技术用于确定样品的基因突变及其基因型 ;RT- PCR半定量法分析该突变对 β-珠蛋白基因转录水平的影响。　结果　家系分析发现 :先证者、先证者之兄和其母亲为有典型 β-地贫特征的个体 ,反向点杂交筛查未发现已知的 β-地贫突变 ,DNA直接测序分析发现该家系的这 3个成员均携带有 1种中国人中未见报道的 β地贫等位基因—— β-珠蛋白基因启动子区近侧 CACCC盒中 - 90位的 C→ T转录突变。RT- PCR分析显示该突变引起β-珠蛋白基因转录水平下降 (突变 :2 .2 33± 0 .0 1vs正常 :3.779± 1.19;95 % CI:3.0 6 0 ,4 .4 99) ,与其他类型的β-地贫杂合子β-基因的表达水平相当 (2 .110± 0 .5 3,95 % CI:1.732 ,2 .4 88)。　结论　 - 90位的 C→ T转录突变是中国人中未见报道的稀少类型的 β-地贫转录突变。     

江西地区β地中海贫血基因突变类型的分析

目的　探讨江西地区 β地中海贫血基因突变类型及其频率。方法　应用聚合酶链反应结合反向点杂交技术分析了 35例重型β地贫患儿及双亲 136条染色体的β珠蛋白基因。结果　检测到 6种突变类型 :IVS2nt- 6 5 4 (C→T)、CD4 1- 4 2 (-TCTT)、CD17(A→T)、TATAbox - 2 8(A→G)、CD2 7- 2 8( C)、CD71- 72 ( A)。结构比依次为 0 3917、0 2 794、 0 12 5、 0 1176、 0 0 5 15、 0 0 2 94。结论　研究结果对在该地区开展遗传咨询、基因分析及产前诊断具有重要意义。     

广东省珠海市户籍人群中β-地中海贫血的分子流行病学调查

目的　调查珠海市这座新兴“移民”城市中户籍人群中 β-地中海贫血 (β -地贫 )的携带率、基因突变类型及其频率分布 ,包括 β -地贫合并α -地贫的发生率。方法　连续抽取婚前对象的静脉血大样本 ,先用血液学分析法筛查 β -地贫表型阳性样品 ,再应用反向点杂交 (RDB)和gap -PCR技术分别对之进行β -地贫基因和东南亚缺失型 (- - SEA)α-地贫的定型 ,未知样品则进行DNA直接测序分析。结果　在 1,5 84例婚检对象中 ,检出 β -地贫表型阳性样品 37例 ;经基因分析 ,34例被确定了β -地贫基因型 (包括 2个小红细胞症且 3 5 %≤HbA2 <4 .0 %的病例 ) ,共检出 9种基因型并总结出它们的构成比。其余的 3例 (有 2例 3 5≤HbA2 <4 .0 %)经DNA测序未发现突变。因此 ,该市户籍人群中的 β -地贫基因携带率为2 2 %(35 / 1,5 84 ) ;9种已知β -地贫的基因频率—CD4 1- 4 2s(-CTTT)和 - 2 8(A→G)分别为 34 3%和 2 5 7%;IVS - 2nt6 5 4(C→T)和CD2 6 (G→A ,BE)、CDs71- 72 (+A)t和CD4 3(G→T)各为 8 6 %、5 7%;CD17(A→T)、- 2 9(A→G)和CDs2 7- 2 8(+C)及未知突变均为 2 85 %。值得提及的是 ,在 35例 β -地贫基因阳性样品中 ,有 2例是从 3 5 %≤HbA2 <4 .0 %样品中检出。此外 ,还从 35例 β-地贫基因阳性样品     

贵州西江地区苗族β珠蛋白基因变异

目的对贵州西江地区苗族同胞进行β-地中海贫血基因分析,了解β-珠蛋白基因在苗族人群中的变异情况.方法用常规酚-氯仿抽提法提取β地中海贫血携带者DNA,经PCR-反向点杂交法对β珠蛋白进行突变基因分析,对未检出突变的标本,测定其β-珠蛋白基因序列确定基因变异情况.结果在受检的42例β-地中海贫血携带者中,经PCR-反向点杂交法检出CD41-42突变20例、 CD17 突变17例,未检出突变标本经β-珠蛋白基因序列测定发现了CD2(CAC → CAT)、IVS-2-16(C → G )、IVS-2-74(G → T)、IVS-2-81( C T重合)4种β-珠蛋白基因变异情况.结论贵州西江地区苗族人群中β-珠蛋白基因变异很有自己独特的特点.     

β-地中海贫血三重突变杂合体的基因型分析

目的 对1例罕见的β-地中海贫血三重基因突变杂合子进行基因型分析.方法 采用反向点杂交和DNA测序法分析患者β-珠蛋白基因型.结果 反向点杂交结果显示,此病例β-珠蛋白基因上有转录子+40～+43、密码子41/42、IVS-2-654 3种突变.基因克隆以及DNA测序分析证实患者一条染色体上存在+40～+43(-AAAC)+CD41/42(-TCTT)复合突变;其同源染色体上存在IVS-2-654(C→T)点突变.患者基因型为[+40～+43(-AAAC)·CD41/42(-TCTT)]/IVS-2-654(C→T).结论 β-地中海贫血[+40～+43(-AAAC)·CD41/42(-TCTT)/N]突变基因型是我国一种新的地中海贫血突变类型.     

深圳地区β-珠蛋白生成障碍性贫血基因突变分析

目的对深圳地区β-珠蛋白生成障碍性贫血患者的β-珠蛋白基因序列进行分析,了解深圳地区人群β-珠蛋白基因的突变类型。方法收集100例深圳地区β-珠蛋白生成障碍性贫血患者的外周血并抽提基因组DNA,通过聚合酶链反应扩增全长β-珠蛋白基因,经DNA测序确定β-珠蛋白基因的突变类型。结果在89例深圳地区β-珠蛋白生成障碍性贫血患者中,我们在β-珠蛋白基因中共发现7种突变,以突变频率高低依次为IVS-Ⅱ-654(C→T)、Codon17(A→T)、Codon41/42(-TTCT)、Codon71/72(+A)、Codon27/28(+C)、Codon43(G→T)、-28(A→G)。剩余11例临床诊断的β-珠蛋白生成障碍性贫血患者的β-珠蛋白基因未检出突变。结论在深圳地区人群中IVS-Ⅱ-654(C→T)、Codon17(A→T)和Codon41/42(-TTCT)为β-珠蛋白基因的常见突变。不排除未检出突变患者存在β-珠蛋白基因座控制区存在突变,这将给产前诊断带来困难。     

常染色体隐性遗传性早发型帕金森综合征DJ1基因突变研究

目的探讨常染色体隐性遗传性早发型帕金森综合征（autosomal recessive early-onset Parkinsonism，AR—EP）DJ1基因的突变特点。方法 应用聚合酶链反应结合DNA直接序列分析方法，对11个常染色体隐性遗传性早发型帕金森综合征家系先证者的DJ1基因进行突变研究。结果本组AR-EP患者未发现DJ1基因的致病突变，在内含子区发现6个多态，分别为IVS1→15T→C、IVS4＋30T→G、TVS4＋45G→A、IVS4＋46G→A、IVS5＋31G→A和g．168-185del，其中3个（IVS1-15→C、IVS4＋45G→A、IVS4＋46G→A）为新发现的多念。结论中国人常染色体隐性遗传性早发型帕金森综合征患者DJ1基因突变可能罕见。     

β地中海贫血杂合子基因突变及Gγ珠蛋白基因-158位点SNP与Hb F的关系

目的　探讨 β地中海贫血 (简称 β地贫 )杂合子基因突变类型和 Gγ珠蛋白基因启动子 - 15 8位点 (Gγ- 15 8)单核苷酸多态性与胎儿血红蛋白 (fetal hemoglobin,Hb F)水平的关系。方法　抗碱 -比色法测定 Hb F水平 ;PCR-寡核苷酸斑点杂交法检测β地贫基因型 ;限制性内切酶 Xmn 消化经 PCR扩增的Gγ基因启动子 DNA片段 ,分析Gγ- 15 8位点的单核苷酸多态性。结果　 6 3例受检的轻型β地贫中 15例 Hb F≥ 2 % (2 .0 6 %～ 10 .4 4 % )。共检出 6种β地贫基因突变 ,分别是 :CD4 1/42 (- TTCT)、CD17(A→T)、nt- 2 8(A→ G)、CD71/72 ( A)、IVS- II- 6 5 4 (C→ T)、IVS- I- 1(G→ T)。 CD4 1/42、CD17、CD71/72、IVS- II-6 5 4的杂合子在 15例 Hb F升高组和 4 8例 Hb F正常组各自所占比例相同。 6 3例个体中有 10例为Gγ-15 8(C→T)突变的杂合子 ,总检出率为 15 .9% ;其中 15例高 Hb F个体中检出 8例 (检出率 5 3.33% ) ,HbF正常的 4 8例检出 2例 (检出率 4 .17% ) ,两组检出率差异有显著性 (P<0 .0 0 1)。结论　 β地贫基因突变CD4 1/42、CD17、CD71/72、IVS- II- 6 5 4与 β地贫杂合子的 Hb F水平无关 ;而 Gγ- 15 8(C→ T)突变与广西地区 β地贫杂合子 Hb F升高密切相关。     

长QT综合征 KCNQ1基因突变筛查方法

目的 研究中国人长QT综合征(long QT syndrome，LQTS)与编码缓慢激活延迟整流钾通道基因(postassium voltage—gated channel，KQT—like subfamily member 1，KCNQl)突变的关系。方法 根据心电图T波的特征对31个家系进行基因分型的初步预测。对10个预测为LQTl家系的家庭成员，用聚合酶链反应-单链构像多态性(polymerase chain reaction—single strand conformation polymorphism,PCR—SSCP)方法进行KCNQ1基因16个外显子及剪接位点的筛查，SSCP异常者进行DNA测序。为避免遗漏心电图表现不典型的LQT1，同时也为了进行方法学比较，对其它21个非LQT1家系只对先证者进行16个外显子的PCR和DNA直接测序。对测序有异常者，分析其家系成员相应外显子的疾病分离情况。若异常只存在于患者，则检查该异常在50个正常对照者中的情况。结果 (1)在心电图预测分型为LQT1的家系中发现了位于第5外显子的S277L(1个家系)和G306V(1个家系)2个错义突变。另外发现了3个多态性，分别为435C→T(1145I)(7个家系)、1632C→A(S546S)(1个家系)、IVS1 9C→G(3个家系)。(2)在心电图分型预测为非LQT1的先证者中只发现了1个剪接突变IVS1 5G→A(2个家系)和1个多态性IVS1 18C→T(1个家系)。3个突变位点均位于KCNQ1基因功能区域，各突变家系内患者存在同样的异常条带或序列，而正常对照无此异常。结论 在中国人LQTS患者中发现TKCNQ1基因上的2个新错义突变、1个剪接突变和4个多态性。结合心电图分型预测，PCR-SSCP法可发现绝大部分突变，是筛查LQTS突变的简便而经济的方法。     

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.     

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 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.