长链非编码 RNA Gm15577参与小鼠小脑神经元增殖与分化

目的鉴定新的参与奈美亨综合征（ NBS ）小鼠模型中与小脑发育缺陷相关的长链非编码RNA（ lncRNA）,并探讨其与小脑增殖分化相关的功能。方法利用芯片技术分析野生型（野生组,Nbs1CNS-ctr）小鼠及Nbs1神经元特异性敲除（突变组,Nbs1CNS-del）小鼠小脑中RNA的表达差异;利用即时荧光定量PCR技术对芯片结果加以验证;针对目的RNA,检测其全身表达及小脑中表达模式、细胞内定位以及对母基因的表达调控模式。结果 lncRNA Gm15577是由神经生长调节因子-1（ Negr1）基因内部的内含子区反向转录而成,特异性表达于小脑,且在其发育过程中呈动态变化趋势,而Nbs1缺失之后整体表达水平显著降低;同时,与未分化期比较,分化后的小鼠小脑原代神经前体细胞中Gm15577及其母基因Negr1均有显著升高;Gm15577敲低之后导致Negr1以及小脑增殖分化相关因子Shh与β-catenin 在RNA水平的表达显著下调;人髓母细胞瘤临床样本数据分析表明Negr1基因在正常人群与肿瘤患者之间有显著的表达差异。结论从神经特异性敲除Nbs1小鼠模型出发报道了一个新的与小鼠小脑发育相关的lncRNA Gm15577,初步结果表明Gm15577通过调控Negr1进而影响小脑神经元增殖与分化进程中的Shh与β-catenin信号通路,其基因行为异常有可能与髓母细胞瘤发病有一定的相关性。     

胶质瘤中PTB调控长非编码RNA选择性剪接的筛选

目的探究在神经胶质瘤发生发展过程中PTB调控选择性剪接的长非编码RNA。方法全转录组芯片分析筛选受PTB蛋白介导影响选择性剪接的长非编码RNA。提取胶质瘤和正常细胞系及临床病例组织样品和敲低PTB的神经胶质瘤细胞总RNA,实时荧光定量PCR验证候选基因不同剪接异构体的表达及比值变化。核质分离实验鉴定长非编码RNA细胞定位。结果筛选出16条受PTB蛋白介导影响选择性剪接的长非编码RNA,其中linc00882的不同剪接异构体的表达模式受PTB调控所产生的差异最明显。检测到linc00882定位于细胞质。结论神经胶质瘤中PTB蛋白参与长非编码RNA linc00882的选择性剪接。     

长链非编码RNA与胃癌的关系研究新进展

长链非编码RNA（10ngnon—codingRNA,LncRNA）是调节性非编码RNA之一,转录本长度〉200nt,表达具有时空特异性,无明显可读框,不编码蛋白质。在细胞质,会影响RNA稳定性和miRNA活性;在细胞核,主要调控基因转录和mRNA剪接。根据与最近编码蛋白质基因的关系,LncRNA可分为五类：     

LncRNA Gm13568调控A1型星形胶质细胞活化及在EAE小鼠病变中的作用

目的:探讨长链非编码RNA(long non-coding RNAs,lncRNA)Gm13568对A1型星形胶质细胞活化及实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠疾病进程的影响与分子机制。方法:构建特异性靶向星形胶质细胞的lncRNA ...     

长链非编码RNA H19与肿瘤关系的研究进展

长链非编码RNA(LncRNA)H19与肿瘤关系密切。H19在肺癌、乳腺癌、胃癌中起促癌作用,在肝癌中则具有双向作用。H19的作用机制有组织特异性,各不相同,多与微小RNA、影响原癌基因表达、EMT形成及甲基化状态下IGF2的印迹状态等有关,是一种有价值的潜在肿瘤生物靶标。     

YTHDC1基因在小鼠小脑发育及人髓母细胞瘤组织中的动态表达

目的探讨RNA m~6A甲基化结合蛋白(YTH结构域包含蛋白1,YTHDC1)在小脑发育和髓母细胞瘤中的表达模式及其可能发挥的作用。方法分别以小鼠小脑、人髓母细胞瘤和细胞系为研究对象,在转录组测序结果的基础上,分析RNA水平表达模式;用免疫印迹和免疫组化检测YTHDC1蛋白的表达;在Daoy细胞中敲低甲基转移酶METTL3并通过m~6A-IP-qPCR方法检测YTHDC1 RNA甲基化水平及对YTHDC1蛋白丰度的影响。结果小鼠小脑发育过程中YTHDC1 RNA水平未见明显改变,蛋白表达显著下降(P0.05)。YTHDC1在P7的小脑内颗粒神经元高表达,而在P60小脑中表达降低。与正常或癌旁组织相比,YTHDC1在肿瘤组织中RNA水平未见明显改变,蛋白水平总体偏低(P0.05)。YTHDC1在髓母细胞瘤中RNA m~6A甲基化水平显著升高(P0.05),而METTL3敲低引起YTHDC1 m~6A甲基化水平下降以及蛋白表达上调。结论小鼠小脑发育及人髓母细胞瘤中YTHDC1蛋白表达变化显著,这一过程受到m~6A甲基化介导的转录后调控作用,进而影响小脑发育及髓母细胞瘤的发生。     

降钙素/降钙素基因相关肽基因前体RNA的加工

降钙素和降钙素基因相关肽是由一个基因表达出的两种功能完全不同的多肽激素,其基因的表达主要在RNA水平上受到调控。前体RNA在加工成为成熟RNA的过程中,在不同的组织中经过不同的剪接形成不同的RNA。其剪接的特异性受到顺式元件和反式因子的调控,基因 的exon4上游剪接受点和下游polyA位点均存在弱的加工位点,通过增强子序列使之产生选择性剪接,另外,可能存在组织特异的反式剪接因子。     

Nbn基因敲除小鼠小脑形态发育的变化

目的:通过观察神经特异性Nbn基因敲除(Nbn-CNS-del)和对照(Nbn-CNS-Ctr)小鼠小脑发育的形态学变化,探讨DNA损伤修复基因Nbn在小鼠小脑发育过程中的作用.方法:应用光镜连续切片和体视学方法,对出生后(P)7、 14、 21d的实验组和对照组小鼠小脑进行系统地形态学观察和定量分析.结果:Nbn-CNS-del小鼠小脑发育比Nbn-CNS-Ctr明显滞后,至P21小脑截面积约减少4倍、小脑小叶截面积约减少6倍;外颗粒细胞层截面积、体密度和小脑小叶的体密度差异有统计学意义.结论:DNA损伤修复基因Nbn是小鼠小脑发育的一个关键基因,明显影响小鼠小脑的发育与成熟.     

人脂肪干细胞脂向分化中长非编码RNA的差异表达

背景：肥胖导致了包括高血压、冠心病、脂肪肝、高脂血症、2型糖尿病在内的诸多疾病,因此深入理解脂肪细胞分化机制对于肥胖的防治具有深远意义。目前对于脂向分化机制的研究多集中在微小RNA上,对于长非编码RNA在脂向分化过程中的作用尚且知之甚少。 目的：获得脂向分化过程中差异倍数明显的长非编码RNA,并进一步筛选在脂向分化过程中可能发挥重要作用的长非编码RNA进行验证。 方法：取人腹部皮下脂肪,采用组织块培养法获取脂肪干细胞,传至第3代后进行成脂诱导分化,通过微阵列技术对脂向分化过程中0,5,12 d长非编码RNA及mRNA差异性表达量进行统计,并结合生物信息学报告筛选出呈现明显差异性表达的长非编码RNA,通过qRT-PCR进行验证。 结果与结论：脂向分化过程中以差异倍数1.5(P < 0.05)为标准,上调长非编码RNA的数量5 d vs. 0 d 748个,12 d vs. 0 d 847个,12 d vs. 5 d 593个;下调长非编码RNA的数量5 d vs.0 d 828个,12 d vs.0 d 1 113个,12 d vs.5 d 750个;结合生物信息学分析结果,在与脂类代谢有关的28个长非编码RNA中根据诱导0,5,12 d差异表达倍数较高且其靶基因可能是已知的与成脂相关的基因中,筛选出3个长非编码RNA：AK304548、BP216319、DA852857。通过PCR验证结果显示AK304548、BP216319及其靶基因表达量呈现先下调后上调的趋势,与微阵列测序结果相符,预示其在脂向分化过程中起到调控作用。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

基因间长非编码RNA:T-UCRs转录活性的验证

目的选择小鼠大脑皮层胚胎发育阶段转录的一类非编码RNA,即基因间区的T-UCRs作为研究对象,探究小鼠基因组上基因间超保守区域的转录活性。方法 1)利用UCSC数据库(NCBI37/mm9)通过生物信息学预测筛选在小鼠14.5 d胎脑中可能表达的基因间UCR;2)Coding Potential Assessment Tool(CPAT)和Coding Potential Caculator(CPC)对筛选结果进行编码能力预测;3)RT-PCR对筛选结果进行验证;4)Real-time PCR分析T-uc.62在不同组织中的表达差异以及在小鼠不同发育阶段脑组织中的表达变化。结果 1)通过UCSC网站上的RNA-seq数据库筛选得到16个可能在小鼠E14.5脑组织中表达的基因间UCRs;2)通过CPAT和CPC分析,16个UCRs均不具有蛋白编码能力,提示可能是非编码RNA;3)经过RT-PCR验证发现有15个是可以表达的;4)以T-uc.62为例,其主要在小鼠发育阶段的脑组织中高表达,而在小鼠的成年组织中低表达甚至不表达。结论基因间超保守区域可以转录生成长非编码RNA,其中,T-uc.62主要在小鼠发育阶段的脑组织中高表达;其可能在大脑发育过程中发挥重要功能。     

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.