DNA 条形码技术在黑瞎子岛鼠类鉴定中的应用

为了探索基于COⅠ基因的DNA条形码技术在鼠类鉴定中的应用,本研究检测了黑瞎子岛地区2科3属4种41只鼠类的COⅠ基因序列,进行比对分析,基于Kimura双参数模型计算种内种间遗传距离,用NJ法构建系统发育树,结果表明种间遗传距离显著大于种内遗传距离,聚类分析表现为很短的种内分支和较长的种间分支,同种个体聚为高支持度的单一分支。5只红背Clethrionomys rutilus 被误判为棕背 Clethrionomys rufocanus,24只莫氏田鼠Microtus maximowiczii被误判为东方田鼠Microtus fortis,1只大林姬鼠Apodemus peninsulae被误判为东方田鼠。研究结果表明, DNA条形码不仅能够纠正形态学鉴定中的错误,还能鉴别形态相近的近缘种。基于COⅠ基因的DNA条形码技术能够方便、快捷、准确的鉴定鼠类。     

六种蚊虫DNA条形码序列分析

传统分类学鉴定需要有完整无残缺的成虫标本,对于非昆虫分类学研究人员应用传统分类学方法鉴定蚊虫较为困难;本研究旨在发掘更多适用于蚊类鉴定的DNA条形码,实现口岸一线蚊类的快速准确鉴定。本文通过提取14个蚊虫样品的基因组DNA,使用通用引物PCR扩增ITS1、ITS2、Cytb、COI基因及序列,PCR产物双脱氧法测序,结合NCBI中相关数据分析部分ITS2、Cytb、COI序列的相似性和遗传关系。结果显示:获得了14个蚊虫样品的部分ITS1、ITS2、Cytb、COI序列的非全长碱基序列共计39条;依据种内遗传距离和相似度将14个未知蚊虫分子分类鉴定为6种蚊虫:凶小库蚊、致倦库蚊、尖音库蚊、赫坎按蚊、白纹伊蚊、里海伊蚊;碱基使用率统计表明所测样品的Cytb、COI基因符合昆虫线粒体DNA碱基组成;构建基因ITS2、Cytb、COI系统进化树,符合蚊虫系统发育遗传进化理论。本研究对14个未知蚊虫样品的进行了分子鉴定,为蚊类的快速准确识别鉴定提供基础数据。     

基于线粒体COⅠ基因序列进行库蠓亚属(双翅目:蠓科)近似种的分子鉴定

为了更快速、准确地鉴别库蠓近似种以及弥补传统形态学在种类鉴定中存在的操作繁琐、难度大等不足,本文采用DNA测序的方法获得库蠓亚属的刺螫库蠓Culicoides punctatus、灰黑库蠓C.pulicaris和新替库蠓C.newsteadi等3种库蠓近似种的部分线粒体COⅠ基因序列,并对其进行分子鉴定;基于Kimura 2-parameter模型分析遗传距离,同时应用MEGA 6.0软件构建系统发育树。结果显示,序列经过Clustal W比对及人工校对后,上述3种库蠓的COⅠ序列长度为394 bp;遗传距离在种内和种间具显著差异(P﹤0.05);系统发育树中不同库蠓种类各自构成单系(群),同种类不同地理种群聚为一支。本研究初步证实了线粒体COⅠ序列可应用于库蠓近似种的分子鉴定。     

DNA条形码研究进展

DNA条形码（ DNA Barcoding ）技术是一种新的物种识别方法,它是分子生物学和生物信息学相结合的产物。在最近几年里,该技术已成为生物分类学中引人注目的研究热点。这一概念认为,类似于商店里使用扫描仪读取条形码,对地球上每一种生物通过快速分析其DNA中的一段基因（线粒体细胞色素c氧化酶Ⅰ亚基, mt COⅠ）加以识别。理论上, DNA条形码已被证明在生物分类鉴定中具有非常重要的作用,并推动了一系列相关学科的发展,但目前不同分类学家对其持的意见也不尽相同。本文综述了DNA条形码技术的产生、发展概况、原理与操作及其在分类中的应用,并概括了DNA条形码在应用于物种分类中可能存在的问题。     

蚊虫线粒体DNA限制酶片段长度差异（RFLDs）分析

中华按蚊，致倦库蚊和五个地理株的白蚊伊蚊线粒体DNA分别甩内切酶消化，其片段在琼脂糖凝胶电泳上观察并做限制酶片段长段差异分析(RFLDs)。结果表明，不同种蚊虫的线粒体DNA存在着显著的片段长度差异，揭示其高度的遗传分叉；而在同种不同地理株的白蚊伊蚊之间，用RFLDs分析所见差异报少，仅在广州株与其它四株之间存在一定的差异。本文对RFLDs分析技术问题也进行了探讨。     

DNA条形码鉴定技术在蚋科昆虫鉴定中的应用进展

蚋是一种重要的医学与畜牧学昆虫,是人类盘尾丝虫病和河盲症的传播媒介。不同种类的蚋在形态学上表现出高度的相似性,难以用传统形态学方法进行快速准确鉴定。DNA条形码技术作为一门新兴的物种鉴定方法以其灵敏、精确、方便和客观的优势,在蚋科昆虫的分类鉴定中已经得到广泛应用。本文通过介绍DNA条形码的产生、发展和研究现状,探讨其在蚋科昆虫鉴定中应用的技术方法路线,并对DNA条形码技术在蚋科分类研究中的应用前景进行了展望。     

嗜人按蚊rDNA—ITS2基因的克隆鉴定及SNP分析

目的克隆和分析嗜人按蚊核糖体DNA（rDNA）第2内转录间隔区（ITS2）序列，研究嗜人按蚊rDNA-ITS2序列的种属特异性及不同个体rDNA-ITS2序列的单核苷酸多态性（Single Nuclear Polymorphism，SNP）。方法用DNA提取试剂盒从嗜人按蚊中提取DNA摸板，利用蚊虫5．8s和28S序列的保守性设计特异引物进行聚合酶链反应以扩增嗜人按蚊rDNA-ITS2基因，扩增产物经回收纯化后连接到TA载体，经amp^＋LB固体平板筛选的阳性克隆。经amp^＋LB液体培养基培养后进行PCR鉴定、EcoRI酶切鉴定和序列分析。结果经PCR反应扩增出全长556bp的嗜人按蚊rDNA-ITS2基因。纯化后重组克隆经PCB鉴定可重现556bp的特异性条带，其酶切产物亦与目的基因PCR产物位置相同。嗜人按蚊6个克隆的同一性为99．6％，其rDNA-ITS2基因单核苷酸存在颠换和插入，在556的范围内有一个A→T，一个G→T的颠换；一个T的插入。结论嗜人按蚊rDNA-ITS2序列在种间高度保守，但不同个体间也存在一定的SNP多态性。     

rDNA-ITS2序列作为中国按蚊分子鉴别特征的评述:Ⅰ按蚊亚属

对核糖体DNA第2内转录间隔区(rDNA-ITS2)序列作为中国按蚊属按蚊亚属蚊种鉴别的分子特征进行评价.笔者整理和分析了GenBank数据库中注册的所有中国记录的按蚊亚属蚊种的rDNA-ITS2序列,并应用MEGA软件计算其在种内和种间的差异性(p距离).截止2012年10月,GenBank中已注册ITS2序列共584条,隶属按蚊亚属蚊20种(包括同物异名)、待定种5种和杂交种1个.计算结果显示,ITS2序列在种内的变异性小于1％,种间的差异(掣 为1.2％(凉山按蚊与昆明按蚊)～69.5％(待定种YM-2003a与须喙按蚊).另外,发现少数注册(、-矽)列在种内差异性较大,在比对序列时需谨慎.结果提示rDNA-ITS2序列在本研究按蚊中具有良好的种内保守性和种间解析度.     

四种常见蚊虫的CO I基因分子进化分析

为了解无锡4种常见蚊虫的COI基因特征和蚊虫分子系统进化关系,对无锡淡色库蚊、白纹伊蚊、骚扰阿蚊和中华按蚊的COI基因序列进行扩增、测序,在GenBank中对序列进行同源性比对,分析基因特征、颠换率、遗传距离,利用MEGA4．1构建分子系统树。结果显示,4种蚊虫的COI基因序列扩增长度为699～717bp,GC含量为30．62％～32．49％,种间颠换率为11．74％-15．49％,种间遗传距离为0．121～0．151;每种蚊虫为单系群的支持值都为100。COI基因具有种属特异性,可以用于4种常见蚊虫的分类鉴别。     

三峡库区上、下游血吸虫病流行区钉螺线粒体cox1基因遗传变异研究

目的研究三峡库区上、下游血吸虫病流行区钉螺线粒体DNA细胞色素C氧化酶亚单位t（c毗1）基因的遗传变异。方法采集三峡库区上游四川、云南及下游安徽、湖北4省共7个地、市的钉螺样本,提取基因组DNA,PCR特异性扩增线粒体cox1基因并测序,用ClustalX（1．81）软件进行多序列比对,MEGA（4．0）软件Kimura2-parameter法计算遗传距离,邻接法（NJ）和最大简约法（MP）构建系统发生树。结果上游与下游不同地域株钉螺间cox1基因差异约为16％．下游地区的肋壳与光壳钉螺cox1基因差异约为3．7％,上游不同地域株钉螺碱基差异约为5．4％。遗传距离显示,上游四川与云南地域株的遗传距离为0．022～0．050,下游安徽与湖北地域株的遗传距离为0．014～0．027,而上游与下游各地区螺群间的遗传距离在0．127～0．138之间,明显大于上游或下游各地区螺群内的遗传距离。进化树结果表明,下游湖北的荆州、石首和安徽的芜湖、宁国钉螺形成一支系,上游四川的绵竹、新都钉螺同属一支系。但两种方法构建的进化树在云南大理钉螺的归属上存在差异,MP法提示大理钉螺从上游的分支中独立出来,单独形成一类。结论上游与下游不同地域株钉螺cox1基因遗传差异较显著,下游肋壳和光壳钉螺种群内遗传变异较小,而上游光壳钉螺种群内遗传变异较大。     

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.     

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.     

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 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.     

A comparative study of the effects of dimebon,obsidan, finoptin,and cordaron on the functional state of ischemic focus and size of necrotic zone in experimental myocardial infarction

Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996     

Effects of sex steroid hormones on lipid peroxidation and glutathione antioxidant system in rat skin

Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999     

Adhesion and growth of CaCo2 cells on surface-modified PEEK substrata

A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.