喉癌患者血细胞线粒体DNA控制区发现高频率变异

利用PCR SSCP技术检测线粒体DNA(mtDNA)复制控制区中 16 4bp的片段 ,在 2 2例喉癌患者的血细胞中发现 3例同质性突变 ,5例异质性突变 ,而在 12例正常人中未发现带型改变 (0 /12 ) .序列分析发现其中一个样本有T146A ,T199C和T2 0 4C的变异 ,T146A为新发现的线粒体DNA多态性 ;这个研究结果提示血细胞线粒体DNAD Loop区的变异 ,可能与喉癌发生有一定的联系 ,对线粒体DNA突变的更深入的研究可以考虑与细胞内信号传导联系起来 ,这将有助于理解线粒体DNA在实体瘤和恶性血液病的研究以及肿瘤细胞中线粒体DNA的复制机制 ,这对寻找新的肿瘤基因诊断的标志物和监测肿瘤发生的遗传易感性是有意义的 .     

鱼类线粒体DNA控制区的结构和进化:以鳑鱼类为例

以鳑鱼类为例,研究了鱼类线粒体DNA控制区的结构和进化规律.识别了终止序列区、中央保守区和保守序列区3个区域.指出扩展终止相关序列(ETAS)的主体是TACAT和它的反向互补序列ATGTA形成的发夹结构.给出了鱼类中若干重要保守序列的普遍形式.研究结果表明,一般情况下,只有一个行使功能的ETAS,但可能会有多个复制的、不行使功能的ETAS存在.鱼类的保守序列CSB2最为保守.线粒体DNA控制区被认为是由各功能单位形成主体框架,主体框架复制产生重复序列,重复序列产生快速变异,这样造成不同类群间线粒体DNA控制区巨大差异.易突变点和二级结构的存在均可能与变异的发生相关.     

增殖细胞核抗原（PCNA）的分子结构及其生物学功能研究进展

增殖细胞核抗原（PCNA）是真核生物复制复合体的核心成分，具有特殊的环状三级结构. 作为真核细胞DNA聚合酶δ的推动因子，与不同复制相关蛋白结合，协调DNA复制过程. 同时PCNA还作为功能转换因子，通过不同调控方式与多种细胞因子作用，参与了DNA损伤修复、细胞周期调控及凋亡等许多重要的细胞事件. 另外作为细胞增殖的指标，PCNA与肿瘤等细胞增殖性疾病的发生和发展存在相关性，因此在临床上对PCNA的深入研究有重要意义. 文中就PCNA的“功能性”结构及其在不同细胞事件中的功能转换（Function Switch）进行简要综述.     

包囊游仆虫休眠细胞中大核和线粒体DNA的多态性

为了探索纤毛虫休眠细胞中两套遗传系统的作用特征,对包囊游仆虫（Euplotes encysticus）休眠细胞与营养细胞大核DNA和线粒体DNA进行了RAPD比较.结果显示,在所选用的35条随机引物中,包囊游仆虫大核DNA共扩增出220条片段,其中以休眠细胞大核DNA为模板扩增出18条特有片段,以营养细胞大核DNA为模板扩增出44条特有片段,两者存在28%的差异.在所选用的32条随机引物中,线粒体DNA共扩增出154条片段,其中以休眠细胞线粒体DNA为模板扩增出19条特有片段,以营养细胞线粒体DNA为模板扩增出25条特有片段,两者有29%的差异.这些结果表明,包囊游仆虫休眠细胞与营养细胞的大核DNA结构存着一定的差异;两者的线粒体DNA结构也存在差异.因此,包囊游仆虫在休眠细胞形成过程中,大核DNA、线粒体DNA结构可能都发生了一定的变化,并且这些变化可能与休眠细胞形成过程中的形态结构和代谢活动等剧烈变化以及休眠状态下的生理生化变化密切相关.所得结果为揭示纤毛虫细胞结构的分化与细胞遗传物质的作用关系提供了基础资料.     

丙酮酸激酶M2在骨肉瘤中的表达及其生物学功能

目的:研究丙酮酸激酶M2(PKM2)在骨肉瘤组织中的表达水平及其临床各指标的关联性,并初步探讨其中可能的分子机制.方法:通过免疫组化检测PKM2在骨肉瘤组织中的表达水平,SPSS统计软件分析其表达水平与患者临床特征及预后的关系;利用数据库分析骨肉瘤组织中PKM2 mRNA水平与患者生存预后的关系.再通过细胞增殖、克隆形成、Transwell迁移以及小鼠体内成瘤等实验方法观察抑制PKM2对骨肉瘤细胞的增殖、克隆形成、迁移及体内成瘤能力等生物学行为的影响.利用流式细胞术检测PKM2的表达水平与骨肉瘤细胞周期的关系.结果:在骨肉瘤组织中PKM2的蛋白表达水平与肿瘤直径、远处转移、临床分期、患者预后均有较强的相关性(P0.05),即高表达PKM2的肿瘤直径较大(χ~2=5.79,P=0.016)、容易发生远处转移(χ~2=7.30 P=0.007)、临床分期较高(χ~2=8.44,P=0.004),患者预后较差(P0.05).且数据库分析结果也显示,骨肉瘤组织中高表达PKM2 mRNA的患者预后较差(P0.05).在骨肉瘤细胞系中沉默PKM2或应用PKM2的抑制剂均可以显著抑制细胞的增殖、克隆形成及迁移能力(P0.05),以及抑制体内的肿瘤生长速度(P0.05);此外,抑制PKM2可以使CyclinD1表达水平下调进而使细胞周期阻滞.结论:PKM2与骨肉瘤的恶性进展密切相关,可作为骨肉瘤患者独立的预后因子;PKM2可通过调控细胞周期蛋白CyclinD1的表达促进骨肉瘤细胞的增殖.     

冠突伪尾柱虫休眠与营养细胞线粒体DNA的比较

为研究纤毛虫休眠状态下线粒体的行为特征及功能与纤毛虫形成包裹的关系，采用RAPD 技术对冠突伪尾柱虫休眠与营养细胞线粒体DNA进行了比较。结果表明，在选用的15条随机引物中，有5条引物的扩增产物完全相同，其余10条引物的扩增产物各有差异。15条引物共扩增出84条片段，其中共享片段为28条，共享度为66.7％。这说明休眠细胞线粒体DNA 与营养细胞线粒体在很大程度上是一致的，但仍存在一定差异，据此推测细胞在形成包囊的过程中，某些线粒体DNA结构可能发生了改变，这些改变可能与线粒体在包囊内的行为特征及功能有关。     

变包长对WLAN MAC协议性能的影响

为更加准确分析WLAN MAC协议性能,讨论了在数据包长随机分布时的系统吞吐量与RTS门限值及分段门限值的关系.通过二维M arkov链的分析,研究了数据包长度服从均匀分布时的网络吞吐量性能,给出了最佳RTS门限值的计算方法.讨论了分段门限值与系统性能的关系,以及RTS门限值与分段门限值结合对系统性能的影响.计算表明RTS门限值和分段门限值变化时,系统吞吐量性能呈现出复杂变化态势.但存在两者最佳的组合,使得系统吞吐量达到最大值,并给出了近似计算该最佳组合值的方法.最后通过计算机仿真验证了文中的推导结果.     

低氧与运动训练对线粒体自噬影响的研究进展

线粒体是机体主要的供能细胞器,但是氧化损伤等因素会导致线粒体功能失调.研究表明,自噬是线粒体维持自身数量以及质量的动态平衡的主要方式之一.线粒体自噬(mitochondrial autophagy)具有选择性,对受损伤或不需要的线粒体能特异性识别,由自噬体将其包裹并清除.线粒体自噬对维持细胞内环境稳态具有重要意义,是维持整个线粒体网络功能完整性和细胞生存的重要机制之一.运动训练对机体代谢的良性效应可用自噬现象来解释,细胞自噬已成为当前运动科学领域的研究热点.该文主要综述近年来国内外有关低氧与运动训练对线粒体自噬现象影响的研究进展.     

生物活性化合物ML-098的首次合成

正恶性肿瘤的生长过程主要表现为癌细胞的产生、发展、增殖以及侵袭.细胞凋亡是机体重要的生理过程,对维持体内环境稳定起着重要作用.恶性肿瘤的产生和发展与细胞凋亡与增殖的调节失控密切相关,在多步骤肿瘤发生的早期,细胞凋亡异常将导致细胞生长周期延长,促进了变异基因的累积,有助于肿瘤的发生.Rho GTPases家族是公认的调节细胞运动的因子,其主要是通过影响     

异形胞发育的蓝细菌DnaA蛋白的表达研究

为了研究DNA复制与Anabaena sp. PCC7120异形胞发育之间的关系.构建了Anabaena PCC7120的复制起始蛋白DnaA的重组表达质粒,在Escherichia coli中诱导其超量表达,纯化后免疫家兔获得抗体,采用Western blotting检测DnaA在营养细胞和异形胞中的表达情况.结果显示,两种不同类型的细胞中都能够检测到DnaA的表达,而且在不同缺氮诱导时间,异形胞中DnaA的表达存在差异.这说明DNA复制起始与异形胞的发育可能存在着一定的关系.     

Parental origin of mutations of the retinoblastoma gene

Retinoblastoma and osteosarcoma arise from cells that have lost both functional copies of the retinoblastoma gene. Using the cloned retinoblastoma gene and other linked polymorphic loci, it is possible to reconstruct the sequential loss of the two homologous gene copies that precedes the development of these tumours. In non-hereditary tumours, the loss of each of the two homologues occurs somatically; in hereditary cases, the initial mutation is in the germline. Recently, Toguchida et al. reported that the paternally derived copy is preferentially the first one to become mutant during the genesis of non-hereditary osteosarcomas. We report here a similar analysis of patients with retinoblastoma in which we find no such predilection for initial somatic mutations. In contrast, when an initial mutation was a new germline mutation, it was derived from the father, a result which is consistent with new germline mutations arising primarily during spermatogenesis.     

Preferential mutation of paternally derived RB gene as the initial event in sporadic osteosarcoma

Successive loss of function of both alleles of the retinoblastoma susceptibility gene (RB) on human chromosome 13 seems to be critical in the development of retinoblastoma and osteosarcoma. In cases where the tumour is familial and susceptibility is inherited, a mutation in one of the alleles is carried in the germline. We have recently shown that cytogenetically visible germline mutations are usually in the paternally derived gene. Such a bias would not be expected for sporadic (non-familial) tumours, where both mutations occur in somatic tissue, but there has been some indication of a bias towards initial somatic mutation in the paternally derived gene on chromosome 11 in sporadic Wilms tumour. We have now examined 13 sporadic osteosarcomas and find evidence which indicates that in 12 cases the initial mutation was in the paternal gene, suggesting the involvement of germinal imprinting in producing the differential susceptibility of the two genes to mutation.     

基于原子力显微镜研究p53蛋白与PBR322 DNA的相互作用

肿瘤(癌症)是由于细胞在复制过程中,DNA损伤不能修复导致细胞凋亡或者细胞无限增殖而形成的。DNA在细胞中转录和翻译都会涉及蛋白与DNA的结合,肿瘤抑制蛋白也是参与这一过程的关键蛋白之一。然而,众多研究发现,肿瘤抑制蛋白p53具有识别和修复损伤DNA的效果,对于细胞的凋亡、基因的保护和避免癌症发生有着重要的意义。有研究表明,金属镁离子和锌离子可以增强p53蛋白的结构稳定性和p53-DNA的亲和力。因此,我们基于原子力显微镜(AFM),直观地呈现出p53蛋白与PBR322环状DNA相互作用的图像,同时发现p53蛋白可以使环状DNA自身形成聚集或者相交。但是,对于长度相当的5 000bp的线状DNA几乎没有这样的效果,而对于20 000bp DNA不会出现这样的现象。然而,在高浓度镁离子环境下,环状DNA会扭转成为麻花状,即形成超螺旋结构。这一现象,可为p53蛋白功能和作用机理研究提供指导,也为癌症治疗、癌症药物开发以及癌症检测方法提供启发。     

cdx4 mutants fail to specify blood progenitors and can be rescued by multiple hox genes

Organogenesis is dependent on the formation of distinct cell types within the embryo. Important to this process are the hox genes, which are believed to confer positional identities to cells along the anteroposterior axis. Here, we have identified the caudal-related gene cdx4 as the locus mutated in kugelig (kgg), a zebrafish mutant with an early defect in haematopoiesis that is associated with abnormal anteroposterior patterning and aberrant hox gene expression. The blood deficiency in kgg embryos can be rescued by overexpressing hoxb7a or hoxa9a but not hoxb8a, indicating that the haematopoietic defect results from perturbations in specific hox genes. Furthermore, the haematopoietic defect in kgg mutants is not rescued by scl overexpression, suggesting that cdx4 and hox genes act to make the posterior mesoderm competent for blood development. Overexpression of cdx4 during zebrafish development or in mouse embryonic stem cells induces blood formation and alters hox gene expression. Taken together, these findings demonstrate that cdx4 regulates hox genes and is necessary for the specification of haematopoietic cell fate during vertebrate embryogenesis.     

preMiDTM:血浆突变检测技术新平台

随着分子生物学技术的发展,癌症分子特征方面的研究得以不断深入。本文旨在探讨肿瘤相关基因突变的研究发展对癌症早期发现的重要意义,介绍原创preMiDTM血浆循环突变检测技术平台及其临床应用前景。     

Identification of the primary gene defect at the cytochrome P450 CYP2D locus

The mammalian cytochrome P450-dependent monooxygenase system is involved in the metabolism of drugs and chemical carcinogens. The role of these enzymes in toxicological response is exemplified by an autosomal recessive polymorphism at the cytochrome P450 CYP2D6 debrisoquine hydroxylase locus which results in the severely compromised metabolism of at least 25 drugs, and which in some cases can lead to life-threatening side-effects. In addition, this polymorphism, which affects 8-10% of the caucasian population, has been associated with altered susceptibility to lung and bladder cancer. Here we report the identification of the primary mutation responsible for this metabolic defect and the development of a simple DNA-based genetic assay to allow both the identification of most individuals at risk of drug side-effects and clarification of the conflicting reports on the association of this polymorphism with cancer susceptibility.     

基于遗传-神经网络的电机故障诊断

本文采用非线性最小二乘法中的LM(Leven-berg-Marquardt)算法,结合遗传算法进行电机故障诊断的方法,并应用于电机故障的仿真实验,性能明显优于单一的算法结构.1神经网络模型及算法本文采用的LM算法是一种利用标准数值优化技术的快速算法,是高斯-牛顿法的改进形式[1].LM算法网络     

The entropy characters of point mutation

The biological diversity, which depends on the genetic material DNA, is the foundation for a species to survive and evolve. The entropy is the best measurement of biological diversity. Based on the single-parameter and the two-parameter models, here we established some differential equations about the point mutation of a DNA sequence with finite length, as well as some functions describing the processes of the variation in quantities of 4 kinds of bases （A, T, G and C） in the DNA sequence. At the molecular level, we discussed the entropy characteristics of point mutation. The results proved that a species maintained its entropy and evolved in the direction of the increasing biological diversity. In order to testify the theoretical results, we did a series of computer simulations of random point mutation in Matlab environment. The results were well consistent with the theoretical researches.     

Genetic linkage of Werner's syndrome to five markers on chromosome 8.

Werner's syndrome (WS) is a rare autosomal recessive disease in which the affected individuals display symptoms of premature ageing. The substantial phenotypic overlap between WS and normal ageing indicates that these two conditions may have pathogenetic mechanisms in common. The WS mutation has pleiotropic effects, and patients and their cells show many differences compared with normals. Despite extensive study of the clinical and biochemical features of this disorder, the primary genetic defect remains unknown. We have undertaken a genetic linkage study in an effort to identify the locus of the primary defect. Here we report close genetic linkage of the WS mutation to a group of markers on chromosome 8.