微阵列比较基因组杂交分析一例足月小样儿的染色体畸变

目的 分析一例足月小样儿的染色体畸变,探讨患儿低出生体重的原因.方法 采集临床已确诊的足月小样儿外周血并抽提基因组DNA,进行微阵列比较基因组杂交,分析患儿基因组拷贝数的改变.培养患儿及其父母外周血淋巴细胞,进行染色体核型分析并确定患儿染色体畸变的来源.结果 微阵列比较基因组杂交显示患儿在10q125.2→qter区域存在长22 Mb片段的重复,同时在15q26.2→qter区域存在长5 Mb片段的缺失.核型分析显示患儿核型为46,XY,-15,+der(15)t(10;15)(q25;q26)pat.结论 患儿在10q25.2→qter区域存在部分三体,而在15q26.2→qter区域存在部分单体,这两种染色体畸变可能均是导致患儿表现为足月小样儿的病因之一.     

荧光原位杂交联合微阵列比较基因组杂交分析一例原发性闭经患者的染色体畸变

目的分析1例原发性闭经患者的染色体畸变,探讨该患者原发性闭经的可能原因。方法采集临床已确诊的原发性闭经患者外周血,并抽提基因组DNA,进行荧光原位杂交和微阵列比较基因组杂交,分析染色体异常。结果微阵列比较基因组杂交显示患者染色体Xp22.31区域存在长1.637Mb片段的三倍体,Xp21.2-q21.1区域存在长52.156 Mb片段的重复片段。结论微阵列比较基因组杂交技术可以检测染色体微小畸变,值得临床推广应用。     

X/Y染色体间不平衡易位的临床遗传学分析

目的 探讨X与Y染色体不平衡易位患者的遗传效应及其临床特征,并比较各种染色体检测技术的临床应用价值。方法 利用高分辨染色体G显带技术、染色体C显带技术以及微阵列比较基因组杂交技术（array-CGH）对因身材矮小、语言发育迟缓来我院就诊的1例患儿进行染色体检查,并对其家系成员通过G显带核型分析或array-CGH检测分析系谱。结果 患儿染色体G显带结果为46,X,der(X)t(X;Y)(p22.3;q11.21)pat;C显带结果提示异常X染色体末端为异染色质;array-CGH结果提示在X染色体Xp22.33→pter位置发生杂合缺失,片段大小约2.7 Mb,Y染色体Yq11.21→qter位置片段存在,片段大小约14.2Mb。结论 X与Y染色体不平衡易位的患者临床表型具有一定的异质性,X染色体缺失或重复片段较大时可引起女性生长发育迟缓、性腺发育异常及不良妊娠等。高分辨染色体G显带联合array-CGH技术,可提供染色体平衡或不平衡易位的遗传机制、染色体片段来源、大小、所含基因数量及其相应的编码功能信息,对疾病的干预治疗和再发风险评估具有非常重要的意义。     

微阵列比较基因组杂交技术检测先天性心脏畸形胎儿的隐藏基因组拷贝数变异

目的 检测1例先天性主动脉弓离断和室间隔缺损胎儿的基因组拷贝数变异(copy number variations,CNVs),寻找其致病的遗传学证据,探讨微阵列比较基因组杂交技术(array-based comparative genomic hybridization,array-CGH)在分子细胞遗传诊断中应用的可行性.方法 对该患儿脐血及其父母外周血进行常规G显带核型分析,发现胎儿核型为46,XX,t(7;9)(q12;q21),双亲核型正常.进而应用array-CGH芯片对患儿进行全基因组高分辨率扫描分析,采用荧光原位杂交技术(fluorescence in situ hybridization,FISH)对新发现的CNVs进行实验验证.结果 array-CGH分析发现胎儿基因组存在1个病理性亚显微结构的拷贝数变异:del(22)(q11.2)(17 370 128～19 790 009,2.42 Mb).FISH实验结果验证了此22q11.2微缺失的存在.结论 隐藏的22q11.2微缺失可能是此胎儿致病的原因;染色体平衡易位的先天缺陷胎儿可能会含有位于重排断裂点区域之外的亚显微结构基因组拷贝数变异;微阵列比较基因组杂交具有高分辨率、高通量和高准确性等优点,适用于亚显微基因组拷贝数变异的检测.     

微阵列比较基因组杂交技术检测先天性心脏畸形胎儿的隐藏基因组拷贝数变异

目的 检测1例先天性主动脉弓离断和室间隔缺损胎儿的基因组拷贝数变异(copy number variations,CNVs),寻找其致病的遗传学证据,探讨微阵列比较基因组杂交技术(array-based comparative genomic hybridization,array-CGH)在分子细胞遗传诊断中应用的可行性.方法 对该患儿脐血及其父母外周血进行常规G显带核型分析,发现胎儿核型为46,XX,t(7;9)(q12;q21),双亲核型正常.进而应用array-CGH芯片对患儿进行全基因组高分辨率扫描分析,采用荧光原位杂交技术(fluorescence in situ hybridization,FISH)对新发现的CNVs进行实验验证.结果 array-CGH分析发现胎儿基因组存在1个病理性亚显微结构的拷贝数变异:del(22)(q11.2)(17 370 128～19 790 009,2.42 Mb).FISH实验结果验证了此22q11.2微缺失的存在.结论 隐藏的22q11.2微缺失可能是此胎儿致病的原因;染色体平衡易位的先天缺陷胎儿可能会含有位于重排断裂点区域之外的亚显微结构基因组拷贝数变异;微阵列比较基因组杂交具有高分辨率、高通量和高准确性等优点,适用于亚显微基因组拷贝数变异的检测.     

5p部分单体3例全基因组拷贝数变异分析

目的应用全基因组微阵列芯片平台,对染色体核型提示为Cri du chat综合征的新生儿进行全基因组拷贝数变异（CNVs）的检测,以帮助解释基因型与表型的相关性。方法 2009年6月至2010年5月复旦大学附属儿科医院收治的染色体核型提示为Cri du chat综合征的3例新生儿进入研究。采用Cytogenetic Whole-Genome芯片筛查全基因组CNVs,针对发现的所有CNVs进行分析,参照国际基因组拷贝数变异多态性数据库除外正常人群多态性CNVs。结合本研究3例与DECIPHER数据库已报道的Cri du chat综合征患儿的临床表型,行5p缺失大小及范围分析,对重复区域行候选基因分析。结果 3例患儿经微阵列芯片检测,均证实并更为精确的定位了5p的缺失范围。例15p缺失位于5p15.33-p13.3,例2缺失位于5p15.33-5p15.1,例3缺失位于5p15.33-p14.3;此外例2发现9p部分重复,例3发现7p部分重复。结合DECIPHER数据库已报道的5例Cri du chat综合征临床表型,重复区域和候选基因分析显示,临床表型为猫叫样哭声或声音异常：缺失片段重叠区域为5p15.33-15.31内3.86Mb,覆盖（IRX1和IRX2与胚胎形成相关的基因）;临床表型为面容异常：缺失片段重叠区域为5p15.2-15.1内2.51Mb（覆盖ANKH与颅骨干骺端发育相关的基因）。例3合并有先天性巨结肠。因纳入病例均为新生儿,无法评价是否存在智力低下和生长发育迟缓,无法对相应的关键区域进行分析。结论本研究提供了微阵列平台罕见潜在致病可能CNVs的分析方法,进一步为建立5p部分缺失表型基因型关联性提供了依据。     

应用微阵列比较基因组杂交技术对胎儿复杂染色体易位的产前诊断

目的对产前羊水细胞培养染色体核型分析,检测出染色体易位的胎儿应查父母双方染色体,并用基于芯片的微阵列比较基因组杂交(array comparative genomic hybridization,a CGH)技术检测,以明确其易位染色体的来源及胎儿染色体有无微重复或微缺失,探讨微阵列比较基因组杂交(a CGH)技术在检测胎儿染色体异常中的临床价值。方法通过胎儿羊水细胞培养,染色体G显带核型分析,诊断出胎儿染色体异常,核型为46,X,t(X;13;9)(q13;q14;p22),t(3;6)(p13;q23)。对此例标本进行a CGH分析,通过多位点高分辨率扫描确定胎儿染色体有无微重复或微缺失。结果a CGH扫描检测出胎儿染色体在Xq13.1-q13.2(71,699,190-71,820,393)区带存在121kb的缺失,在13q14.2-q21.1(48,706,590-57,520,639)区带存在8.8Mb的缺失。结论利用a CGH技术可以方便快速地鉴定和分析染色体的微重复或微缺失,结合传统的核型分析技术,可以为判断重复或缺失染色体片段的遗传学效应和产前诊断提供帮助。     

罕见的8p部分缺失伴重复患儿1例的临床及遗传学分析

目的明确1例智力低下、语言发育迟缓及自闭症患儿的遗传学病因。方法选取2019年6月30日于泉州市妇幼保健院就诊的1例罕见8p部分缺失伴重复患儿为研究对象。采集患儿及其父母的外周血样, 进行G显带染色体核型分析以及单核苷酸多态性微阵列(SNP-array)检测。结果患儿核型为46, XX, dup(8p?), 其父母均未见明显异常。SNP-array检测显示患儿染色体8p23.3p23.1区存在6.8 Mb的片段缺失, 8p23.1p12区存在21.8 Mb的片段重复, 上述拷贝数变异均为新发, 并判断为致病变异。结论患儿的临床表型与染色体8p部分缺失重复相关, SNP-array检测对智力低下的患儿的诊断具有一定的价值。     

单纯17q25.3拷贝数重复导致全面性发育迟缓和多发性先天异常一例

目的探讨单纯17q25.3拷贝数重复的临床特征、遗传方式及基因型与表型的关系。方法应用全外显子测序、染色体微阵列、染色体核型分析、荧光原位杂交技术联合对先证者及其家系成员进行分析。结果先证者为一例4岁的多发性先天异常男性患儿,表现为全面性发育迟缓、矮小、智力障碍、脑发育不良、小头、特殊面容、肌张力低下、注意力缺陷多动障碍、共济失调、骨骼和心血管异常等。全外显子测序和染色体微阵列分析鉴定其在染色体17q25.3→qter发生5.7 Mb拷贝数重复,可能为患儿致病的原因。荧光原位杂交证实先证者该拷贝数重复是遗传自携带该片段平衡易位的母亲,其外祖母和舅舅也为该片段平衡易位携带者,而小姨未见异常。结论本研究结果丰富了单纯17q25.3拷贝数重复的临床表型谱,为遗传咨询提供了依据,并初步提示了P4HB、ACTG1、BAIAP2及TBCD基因为17q25.3拷贝数重复候选基因。     

Dandy-Walker综合征与7号染色体微缺失

目的 应用微阵列比较基因组杂交技术探讨Dandy-Walker综合征的遗传学病因.方法 选取8例经产前超声检查提示发生Dandy-Walker畸形且常规G显带染色体核型分析未发现异常的胎儿病例,按照标准的Affymetrix cytogenetic 2.7 M微阵列芯片的操作手册进行杂交、洗涤及全基因组扫描,并应用相应的计算机软件分析结果.结果 微阵列比较基因组杂交技术检测提示3例Dandy-Walker畸形胎儿的染色体7p21.3区DNA拷贝数发生了缺失或重复,异常片段中包含与脊髓小脑疾病相关的NDUFA4和PHF14基因.结论 染色体7p21.3区DNA拷贝数的异常改变是Dandy-Walker综合征的病因之一,其发病机制可能与NDUFA4和PHF14基因的表达异常有关.     

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.