一个新生儿肝内胆汁淤积症(NICCD)家系的SLC25A13基因突变检测与产前诊断

目的对一个新生儿肝内胆汁淤积症(neonatal intrahepatic cholestasis caused by citrin deficiency,NICCD)家系的SLC25A13基因突变检测和产前诊断。方法收集患者及父母的外周血标本,提取基因组DNA,在明确先证者病因和基因型的基础上,采用Sanger法对家系中1例已孕15周的胎儿进行SLC25A13基因的相应突变位点进行检测和产前诊断。结果 Sanger法DNA测序检测出该家系中先证者父亲、母亲分别携带SLC25A13基因IVS6+5GA、c.851del4杂合突变,先证者SLC25A13基因IVS6+5GA、c.851del4复合杂合突变来源于父母。对先证者母亲的羊水标本进行此两位点的检测,发现羊水标本SLC25A13基因未携带此两位点的突变,基因型与先证者不一致。结论 SLC25A13基因IVS6+5GA、c.851del4 2个突变为Citrin缺陷导致的NICCD的热点突变,Sanger测序技术可有效的为Citrin缺陷导致的NICCD家系提供遗传咨询和产前诊断服务。     

遗传性耳聋家系的致病原因分析及产前诊断

目的 对3个先天性耳聋家系进行遗传性耳聋基因检测及突变类型分析,为再生育的家庭提供遗传咨询及产前诊断。方法 对在宁波市妇女儿童医院就诊的3例先天性耳聋患儿抽取静脉血,提取DNA。先应用目标基因捕获和高通量测序技术对先证者进行耳聋基因相关的全外显子和相邻内含子测序,利用生物信息学技术对测序数据进行分析,筛选相关变异基因并对其进行致病性分析。再应用Sanger测序法对先证者及其父母做基因突变位点验证。最后对第二胎需要进行产前诊断的孕妇采集羊水进行位点验证。结果 家系1先证者中检出肌球蛋白XVA(myosinXVA,MYO15A)基因c.2075C>A、c.4520G>A、c.6668C>T和c.10250＿10252del杂合突变。家系2先证者中检出MYO15A基因c.5964+3G>A和c.7395+6T>G复合杂合突变。家系3先证者中检出缝隙连接蛋白β2(gap junction protein beta 2,GJB2)基因c.299＿300delAT和c.109G>A复合杂合突变。产前诊断结果显示家系1和家系2胎儿的基因型均与先证者的不同,听力表型...     

遗传性耳聋家系的基因突变分析及产前诊断

目的对两个常染色体隐性遗传耳聋家系进行基因突变分析及产前诊断。方法收集家系成员外周血样本及临床资料,运用PCR产物直接测序技术对家系所有成员进行GJB2、GJB3、SLC26A4、线粒体12S r RNA 4个耳聋相关基因检测,明确耳聋致病基因;结合STR位点分析方法排除产前诊断中胎儿DNA受母体基因组的污染。结果家系1先证者系GJB2基因109GA/235del C复合杂合突变;家系2先证者为SLC26A4 IVS7-2AG/946GT复合杂合突变;产前诊断结果显示家系1和家系2胎儿分别系109GA杂合突变携带者及SLC26A4基因IVS7-2AG杂合突变携带者。结论 GJB2基因109GA与235del C所形成的复合杂合突变可导致重度感音神经性耳聋,DNA片段测序技术有助于寻找耳聋致病基因非热点突变,产前诊断和早期干预能避免耳聋患儿的出生。     

20个希特林缺陷病家系的基因分析及产前诊断CSCD

目的探讨20个希特林缺陷病家系SLC25A13基因的突变特点以及产前诊断的可行性。方法通过高频突变筛查结合直接测序的技术对20例先证者及其父母进行SLC25A13基因突变分析。在确定每个家系基因型后,为先证者母亲再次妊娠的胎儿提供遗传咨询并进行产前诊断。结果 20个希特林缺陷病先证者均检出SLC25A13双等位基因致病性突变,共发现10种致病突变类型,包括3种缺失突变:c.851del4、c.1092;095delT和c.495delA;2种剪接位点突变:IVS6+5G>A和IVS11+1G>A;2种无义突变:c.775C>T （p.Q259X）和c.72T>A（p.Y24X）;1种重复突变:c.1638;660dup;1种插入突变:IVSl6ins3kb;1种错义突变:c.1775A>C（p.Q592P）。20个家系共行24次产前诊断。其中8例胎儿基因型正常,11例为SLC25A13基因突变携带者,5例为SLC25A13双等位基因突变。2例c.851del4/c.851del4纯合突变胎儿的父母选择继续妊娠,其余3例双等位基因突变胎儿的父母选择终止妊娠。结论对希特林缺陷病家系进行SLC25A13基因突变分析,可以为先证者确诊、受影响家庭的遗传咨询和下一胎产前诊断提供实验依据,有效降低缺陷患儿再发风险。     

GJB2基因p.V37I突变致病性及家系临床表型分析

目的探讨GJB2基因突变耳聋家系p.V37I(c.109GA)突变致病性及分析家系患者临床表型。方法收集6个GJB2基因p.V37I(c.109GA)突变耳聋患者及家系的临床资料及外周血样本,运用PCR产物直接测序技术对家系耳聋患者进行GJB2、GJB3基因编码区、SLC26A4基因外显子7和8、以及线粒体m.1494CT、m.1555AG位点检测分析,另对家系5先证者行高通量全外显子序列检测。结果 6个家系均检出GJB2基因p.V37I突变,其中家系3、5为纯合突变,家系1、2、6先证者另复合GJB2基因c.235del C杂合突变,家系4患者另检出c.299-300del AT杂合突变;全部家系均未检出GJB3基因编码区、SLC26A4基因外显子7和8、以及线粒体m.1494CT、m.1555AG位点突变,家系5先证者高通量全外显子序列检测结果亦提示GJB2基因p.V37I纯合突变为其致聋原因。结论 GJB2基因p.V37I突变具有一定致病性,该位点纯合突变可导致轻度至中度听力损失,而p.V37I突变复合c.299-300del AT、c.235del C杂合突变可导致中度至重度感音神经性耳聋。     

三个Fabry病家系GLA基因突变与临床表型

目的 分析3个Fabry病家系GLA基因突变及其与临床表型的关系.方法 应用PCR结合DNA测序技术,检测先证者及相关成员GLA基因编码序列与剪切位点DNA序列变异,分析致病性突变与临床表型关系.结果 在家系1先证者GLA基因第5外显子中发现1个未经报道的错义突变c.797A＞C(D266A),家系2先证者GLA基因第5外显子中发现1个错义突变c.644A＞G(N215S),家系3先证者GLA基因第2外显子中发现1个无义突变c.355C＞T(Ql19X).家系1与家系3先证者主要表现为皮肤损害和慢性肾功能不全,家系2先证者临床则以肥厚性心肌病为特点.结论 首次发现的GLA基因c.797A＞C(D266A)突变是第266位密码子第6个被证实的错义突变,已报道的另5种突变均有致病性,在正常非相关对照中未发现该突变,提示GLA基因c.797A＞C突变很可能是该家系的致病原因.N215S和Q119X系首次发现于中国Fabry病家系的突变.GLA基因不同位点的突变具有较为显著的表型差异.     

鸟氨酸氨甲酰基转移酶缺乏症OTC基因错义突变的分子鉴定

目的 对一个鸟氨酸氨甲酰基转移酶缺乏症(ornithine transcarbamylase deficiency,OTCD)家系进行分子遗传学检测,从基因水平确定其原因,为遗传咨询和产前诊断提供依据.方法 应用聚合酶链扩增技术和Sanger测序法对该家系成员的鸟氨酸氨甲酰转移酶基因(ornithine carbamoyltransferase,OTC)的10个外显子进行直接测序,检测潜在的致病突变,以100名健康人为正常对照.结果 先证者新生儿期发病,OTC基因测序发现其第9外显子发生错义突变c.917G＞C,第306位密码子由AGA突变为ACA,精氨酸替换为苏氨酸,即p.R306T.家系成员检测证实先证者母亲及家系中另外两名女性为表型正常的c.917G＞C杂合突变携带者,其他家系成员及100名对照者未发现上述突变.结论 结合生物信息学分析,错义突变c.917G＞C为该家系的致病原因.该突变尚未见报道,是一新发现的OTC基因突变位点.     

一个非综合征型耳聋家系的分子病因学分析

目的分析一非综合征型耳聋家系的分子病因,为患病家系遗传咨询和产前诊断提供依据。方法采用聚合酶链反应(polymerase chain reaction,PCR)和Sanger测序法对4名耳聋患者进行GJB2和SLC26A4基因编码区及侧翼序列测序。结果先证者GJB2基因序列测定为野生型,检出SLC26A4基因c.240delC和c.2168A>G复合杂合突变。先证者父亲检出SLC26A4基因c.240delC杂合突变,先证者母亲检出SLC26A4基因c.563T>C、c.1746delG和c.2168A>G三个杂合突变,先证者妹妹检出SLC26A4基因c.240delC、c.563T>C和c.1746delG三个杂合突变。其中,SLC26A4c.240delC为未见报道的新发框移突变,其余三个突变均为文献已报道的致病突变。结论该家系先证者、先证者母亲及先证者妹妹均为SLC26A4基因复合杂合突变导致的非综合征型耳聋,先证者父亲仅检出单杂合突变。明确基因突变有助于该家系进行遗传咨询和婚育指导,避免聋儿出生。     

一个X连锁少汗型外胚层发育不良家系的EDA基因c.822G＞T突变分析

目的 对1个少汗型外胚层发育不良家系进行ectodysplasin A(EDA)基因序列分析,为家系成员提供准确病因诊断和遗传咨询.方法 抽取家系中先证者及有血缘关系的家系成员外周静脉血,常规提取基因组DNA,应用聚合酶链式反应、DNA测序技术分析EDA基因编码区序列;选择103名无血缘关系正常人作为对照.结果 在先证者及另1例男性患者的EDA基因第7外显子区域检出c.822G＞T突变,导致第274位的色氨酸变成了半胱氨酸(p.W274C),该突变位点未见文献报道.家系中的5名女性成员中检出c.822G＞T杂合突变,正常男性成员及103名正常对照均不存在此突变.结论 EDA基因c.822 G＞T突变为导致该家系成员少汗型外胚层发育不良的致病突变.     

一个全面性癫痫伴热性惊厥附加症家系临床表型及GABRG2基因突变分析

目的 分析并确定一个全面性癫痫伴热性惊厥附加症(generalized epilepsy with febrile seizures plus,GEFS+)家系临床表型,并对其CABAA受体γ2哑单化基因(GABAA-receptor γ2 subunit,GABRG2)进行突变筛查及遗传特征分析.方法 收集先证者及其家系成员临床资料及外周血DNA,采用聚合酶链反应和DNA直接测序的方法进行GBRG2基因突变筛查,确定基因突变的位点,分析基因型与表型的关系.结果 该家系为典型GEFS+家系,3代共有7例受累成员,临床表型1例为热性惊厥(febrile seizures,FS),6例为热性惊厥附加症(febrile seizures plus,FS+).该家系先证者的GABRG2基因存在第9外显子的杂合无义突变c.1287G＞A(P.W390X),先证者之母和具有GEPS+表型的其他家系成员均携带该基因突变,1例携带该突变的家系成员临床表型正常,外显率约为87.5%(7/8).结论 该GEFS+家系GBRG2基因突变P.W390X为遗传性突变,家系符合常染色体显性遗传伴外显率小全.GABRG2基因突变也是中国GEFS+家系的致病基因之一.     

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.     

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.