Frequent mtDNA mutations and its role in gastric carcinogenesis

In order to disclose the relationship between mutations of mitochondrial DNA (mtDNA) and gastric carcinogenesis, we screened the entire mtDNA sequence in 30 cases of human gastric cancer and matched normal tissues by using denaturing high-performance liquid chromatography (DHPLC) and DNA sequencing. Our data showed that high frequency (66.7%, 20/30) of mitochondrial genome mutation occurred in gastric cancer. Among these variants, 17 cases (56.7%, 17/30) were identified to be somatic mutation. High level mutant frequency was found in ND4, ND5 coding genes and D-loop control region, which was 36.7%, 26.7% and 30% respectively. Comparing with complexes Ⅲ, Ⅳ and Ⅴof the electron transport chain, we found that variants appeared to be more frequent in the subunit genes of complexⅠ. Most of mutations were base substitutions (85.4%, 41/48). Our results suggested that mutations of subunit genes encoding complexⅠ, especially ND3, ND4 and ND5 genes, might contribute to human gastric carcinogenesis.     

裂足轮虫还是裂足臂尾轮虫

通过分析比较了裂足轮虫(Brachionus diuersicornis)、方型臂尾轮虫(B．quadridentatus)、壶状臂尾轮虫(B．urceus)、矩形臂尾轮虫(B．1eydigi)、萼花臂尾轮虫(B．calyciflorus)等5种轮虫的线粒体细胞色素氧化酶亚基I(COI）的部分序列，并结合4种海水臂尾轮虫的序列数据，用西氏晶囊轮虫(Asplanchna．sieboldi)作外群构建UPGMA树和NJ树，探讨了烈足轮虫的分类学问题，认为将裂足轮虫归属千臂尾轮虫属市为合适。     

分子系统学及其应用

本文从生物多样性的研究与保护角度，介绍了分子系统学的产生与发展以及分子系统学中的研究方法。     

Transient and constitutive repression of cytoplasmic translation signaling in cells with mtDNA mutation

Cytoplasmic translation is under sophisticated control but how cells adapt its rate to constitutive loss of mitochondrial oxidative phosphorylation is unknown. Here we show that translation is repressed in cells with the pathogenic A3243G mtDNA mutation or in mtDNA-less ρ⁰ cells by at least two distinct pathways, one transiently targeting elongation factor eEF-2 and the other initiation factor eIF-2α constitutively. Under conditions of exponential cell growth and mammalian target of rapamycin (mTOR) activation, eEF-2 becomes transiently phosphorylated by an AMP-activated protein kinase (AMPK)-dependent pathway, especially high in mutant cells. Independent of AMPK and mTOR, eIF-2α is constitutively phosphorylated in mutant cells, likely a signature of endoplasmic reticulum (ER)-stress response induced by the loss of oxidative phosphorylation. While the AMPK/eEF-2K/eEF-2 pathway appears to function in adaptation to physiological fluctuations in ATP levels in the mutant cells, the ER stress signified by constitutive protein synthesis inhibition through eIF-2α-mediated repression of translation initiation may have pathobiochemical consequences. Received 29 October 2008; received after revision 11 December 2008; accepted 16 December 2008     

青鱼线粒体DNA限制性酶切图谱的研究

用10种限制性内切酶对青鱼(Mylopharyngodonpiceus)的肝脏mtDNA进行了分析,其分子质量为9 949×106u,分子大小约为16 60kb.PstⅠ、EcoRⅠ、XhoⅠ、HindⅢ、BamHⅠ、PvuⅡ、SalⅠ、XbaⅠ、BglⅡ、BglⅠ在青鱼的mtDNA分子上分别具有0、3、1、4、4、3、2、2、2、3个切点.根据单酶解及双酶解结果建立了青鱼mtDNA的限制性酶切图谱.     

检测线粒体DNA SNPs的快速测定法--引物延伸-飞行时间质谱法

用PCR扩增出人类mtDNA HV Ⅰ区443bp片段作为模板,以nt16093位点特异的引物进行引物延伸反应。产物经纯化后利用MALDI-TOF质谱技术进行检测,从而建立起引物延伸—飞行时间质谱法快速检测SNPs的技术,并用该法对人类mtDNA HV Ⅰ区SNPs位点nt16093的T／C多态性进行频率调查。结果为广州地区汉族人群的mtDNA HV Ⅰ区多态位点nt16093T型的频率为89．6％,C型的频率为10．4％。并且发现3例异质性。     

Pitfalls in the analysis of ancient human mtDNA

The retrieval of DNA from ancient human specimens is not always successful owing to DNA deterioration and contamination although it is vital to provide new insights into the genetic structure of ancient people and to reconstruct the past history. Normally, only short DNA fragments can be retrieved from the ancient specimens. How to identify the authenticity of DNA obtained and to uncover the information it contained are difficult. We employed the ancient mtDNAs reported from Central Asia (including Xinjiang, China) as an example to discern potentially extraneous DNA contamination based on the updated mtDNA phylogeny derived from mtDNA control region, coding region, as well as complete sequence information. Our results demonstrated that many mtDNAs reported are more or less problematic.Startim, from a reliable mtDNA phylogeney and combining the available modern data into analysis, one can ascertain the authenticity of the ancient DNA, distinguish the potential errors in a data set, and efficiently decipher the meager information it harbored. The reappraisal of the mtDNAs with the age of more than 2000 years from Central Asia gave support to the suggestion of extensively (pre)historical gene admixture in this region.     

成都麻羊与四川各地黑山羊品种(群体)mtDNA D-1oop序列多态性研究

采用mtDNA多态性标记对成都麻羊与四川各地黑山羊品种(群体)D-loop序列多态性进行分析.结果表明,供试山羊品种(群体)mtDNA片段长度为16kb左右,mtDNA D-loop片段长度为1210～1212bp,共检测到10种单倍型,群体间共有多态座位83个,单一多态座位23个,简约信息座位24个,平均核苷酸歧异度为0.001510,D-loop序列多态性较贫乏.经聚类,成都麻羊与四川各地黑山羊品种(群体)分为两大类,成都麻羊首先与金堂黑山羊(D=0.203)聚在一起后,再依次与乐至黑山羊(D=0.223)、建昌黑山羊(D=0.477)、白玉黑山羊(D=0.593)形成一大类;合江黑山羊与江安黑山羊(D=0.231)先聚在一起后,再与自贡黑山羊(D=0.337)聚为1类,营山黑山羊与嘉陵黑山羊(D=0.242)聚为1类,2类形成另一大类.最后两个大类聚在一起.聚类结果与供试山羊品种(群体)来源和生态地理分布相一致.     

大熊猫线粒体DNA酶切片段长度多态性研究

用１２种限制性内切酶（ＢａｍＨⅠ，ＥｃｏＲⅠ，ＥｃｏＲⅤ，ＸｈｏⅠ，ＰｓｔⅠ，ＭｓｐⅠ，ＸｂａⅠ，ＰｖｕⅡ，ＨｉｎｄⅢ，ＨａｅⅢ，ＳａｃⅠ，ＨｐａⅡ）对秦岭大熊猫线粒体ＤＮＡ进行了酶切。测定和分析了每种酶所产生片段的数量和大小，并与四川大熊猫的限制性片段进行了比较，发现在相同的６种酶的１６个酶切位点中有一个不同，表明这两地群体间存在着线粒体ＤＮＡ的多态性。其研究结果为进一步研究和保护大熊猫提供了重要依据。     

MGB probe assay for rapid detection of mtDNAl1778 mutation in the Chinese LHON patients by real-time PCR

Objective： Leber＇s hereditary optic neuropathy （LHON） is a maternally inherited degeneration of the optic nerve caused by point mutations of mitochondrial DNA （mtDNA）. Many unsolved questions regarding the penetrance and pathophysiological mechanism of LHON demand efficient and reliable mutation testing. This study aims to develop a minor groove binder （MGB） probe assay for rapid detection of mtDNA11778 mutation and heteroplasmy in Chinese LHON patients by real-time polymerase chain reaction （PCR）. Methods： Forty-eight patients suspected of having LHON and their maternal relatives underwent a molecular genetic evaluation, with 20 normal individuals as a control group at the same time. A real-time PCR involving two MGB probes was used to detect the mtDNA 1 1778 mutation and heteroplasmy. A linear standard curve was obtained by pUCmLHONG and pUCmLHONA clones. Results： All 48 LHON patients and their maternal relatives were positive for rntDNA11778 mutation in our assay, 27 heteroplasmic and 21 homoplasmic. Eighteen cases did not show an occurrence of the disease, while 9 developed the disease among the 27 heteroplasmic mutation cases. Eleven did not show an occurrence of the disease, while 10 cases developed the disease among 21 homoplasmic mutation cases. There was a significant difference in the incidence between the heteroplasmic and the homoplasmic mutation types. The time needed for running a real-time PCR assay was only 80 min. Conclusion： This real-time PCR assay is a rapid, reliable method for mtDNA mutation detection as well as heteroplasmy quantification. Detecting this ratio is very important for predicting phenotypic expression of unaffected carriers.