西兰花中ITCS诱导SGC-7901细胞凋亡作用机制

研究西兰花中异硫氰酸盐(isothiocyanates,ITCS)诱导人胃腺癌SGC-7901细胞凋亡的作用及其机制.应用MTT法检测ITCS对SGC-7901细胞增殖的影响;流式细胞仪检测活性氧、线粒体膜电位(Δψm)和细胞凋亡率.实验证明ITCS可明显抑制SGC-7901细胞增殖;不同质量浓度的ITCS作用于SGC-7901细胞24 h后,细胞内活性氧含量随给药剂量的增加而升高、线粒体膜电位依次降低.60、120、240μg/mL的ITCS作用于SGC-7901细胞48 h后可见细胞凋亡率分别为9.1%、20.1%和55.4%.ITCS可通过刺激SGC-7901细胞内活性氧的产生,损伤肿瘤细胞线粒体,使其膜电位下降最终引起SGC-7901细胞凋亡.     

Analysis of RAPD and mitochondrial 16S rRNA gene sequences from Trichiurus lepturus and Eupleurogrammus muticus in the Yellow Sea

Random amplified polymorphic DNA (RAPD) technique is applied to 12 individuals from each species of the hairtail fishes Trichiurus lepturus and Eupleurogrammus muticus in the Yellow Sea. The percentage of polymorphic sites, degree of genetic polymorphism and genetic distance are compared and the phylogenetic tree is constructed by Neighbor-joining method. The partial mitochondrial 16S rRNA gene is amplified by polymerase chain reaction (PCR) and the PCR products are directly sequenced after being purified. These sequences, together with the homologous sequences of another Trichiuridae species Lepidopus caudatus obtained from GenBank, are used to analyze nucleotide difference and to construct a UPGMA phylogenetic tree by means of biological informatics. Analysis shows: (1) the RAPD technique is a highly sensitive method for investigating genetic diversity in T. lepturus, and E. muticus. T. lepturus exhibits a lower polymorphism and genetic diversity than E. muticus; (2) according to the analysis of the partial mitochondrial 16S rRNA gene sequences, a very low intraspecific variation and considerably high divergence among species were found, which reveals a dual nature of conservatism and variability in mitochondrial 16S rRNA gene; (3) five primers generate the species-specific RAPD sites and these sites can be served as the molecular markers for species identification and (4) it can be proved at DNA variation level that T. lepturus and E. muticus are of two species respectively pertaining to different genera, which supports the Nelson taxonomic conclusion.     

Complete mitochondrial DNA sequence of Chinese alligator,Alligator sinensis, and phylogeny of crocodiles

The 16746-neueleotide (nt) sequence of mitochondrial DNA(mtDNA) of Chinese alligator,Alligator sinensis,was determined using the Long-PCR and primer walking methods.As is typical in vertebrates,the mtDNA encodes 13 proteins,2 rDNA,22 tRNA genes,and a noncoding control region.The Composition of bases is respectively 29.43%A,24.59%T,14.86%G,31.12% C.The gene arrangement differs from the common vertebrate gene arrangement,but is similar to that of other crocodiles,DNA sequence data from 12S rRNA,168 rRNA,protein-coding genes and combined sequence data were used to reconstruct the phylogeny of reptiles with the MP and ML methods.With this large data set and an appropriate range of outgroup taxa,the authors demonstrate that Chinese alligator is most closely related to American alligator among three crocodilian species,which suppors the traditional viewpoint.According to the branch lengths of ML tree from the combined data set,the primary divergence between Alligator and Caiman genus was dated at about 74.9 Ma,the split between Chinese alligator and American alligator was dated at 50.9Ma.     

Mitochondrial genetic variation within and between Holbrookia lacerata lacerata and Holbrookia lacerata subcaudalis,the spot-tailed earless lizards of Texas

ABSTRACT

We examined genetic relationships among individuals and populations of the species Holbrookia lacerata, the spot-tailed earless lizard, using whole mitochondrial genomes. Lizards were collected from south, central and west Texas. We found significant amounts of genetic structure among populations and evidence of two major reciprocally monophyletic groups of spot-tailed earless lizards in Texas. Holbrookia lacerata lacerata occurs on the Edwards Plateau and adjacent regions of West Texas north of the Balcones Escarpment, while Holbrookia lacerata subcaudalis occurs in South Texas and adjacent Mexico south of the Balcones Escarpment. These two recognised subspecies correspond to the two clades we discovered. Holbrookia l. lacerata occupies much of its historical range at sometimes high population densities, while populations of H. l. subcaudalis appear to be highly fragmented based on recent observations compared to their historical range.     

Mitochondrial association of alpha-synuclein causes oxidative stress

α-Synuclein is a neuron-specific protein that contributes to the pathology of Parkinson’s disease via mitochondria-related mechanisms. The present study investigated possible interaction of α-synuclein with mitochondria and consequences of such interaction. Using SHSY cells overexpressing α-synuclein A53T mutant or wild-type, as well as isolated rat brain mitochondria, the present study shows that α-synuclein localizes at the mitochondrial membrane. In both SHSY cells and isolated mitochondria, interaction of α-synuclein with mitochondria causes release of cytochrome c, increase of mitochondrial calcium and nitric oxide, and oxidative modification of mitochondrial components. These findings suggest a pivotal role for mitochondria in oxidative stress and apoptosis induced by α-synuclein. Received 27 December 2007; received after revision 7 February 2008; accepted 8 February 2008     

An analysis of allozyme,mitochondrial DNA and morphological variation in mussel (Mytilus galloprovincialis) populations from Greece

Three populations ofM. galloprovincialis from northern Greece were investigated using isozyme analysis, discriminant analysis of morphological characteristics and analysis of restriction fragments of mtDNA. For all three types of analysis significant intra- and interpopulation differentiation was found. This differentiation is very noticeable at the mtDNA genotype frequencies. Furthermore, the restriction patterns of mtDNA were different from those reported for Atlantic populations of this species.     

位点特异性频率模型在昆虫分子系统发育分析中的应用

根据位点特异性频率模型(site-specific rate model),利用PAUP软件,对线粒体COI基因序列不同密码子位点的数据模块分别进行了豉甲科和水生肉食亚目在亚科或科水平上的系统发育学分析,结果表明第二密码子数据模块获得的系统分析结果与形态学分析结果一致。表明位点特异性频率模型在线粒体COI基因作为标记来进行昆虫分子系统发育分析上获得了更好的效果。     

An anti-Bcl-2 antibody prevents 2-deoxy-D-ribose-induced apoptosis in the IPLB-LdFB insect cell line

Confocal microscopy reveals that the anti-Bcl-2 antibody (pAb) is able to diffuse across the plasma membrane of the fat body cell line IPLB-LdFB from the insect Lymantria dispar, demonstrating the presence of Bcl-2-like molecules in the cytoplasm. Immunoperoxidase procedure confirms the cellular localization. Furthermore, an immunoprecipitation corresponding to a molecular weight of 29 KDa is observed with western blot analysis using the anti-Bcl-2 pAb. Cytofluorimetric experiments show that anti-Bcl-2 pAb counteracts 2-deoxy-d-ribose-induced apoptosis and provokes morphological changes in the insect cell line, i.e. a reduction in cell size, the disappearance of the vacuola and changes in shape. At the same time, the antibody provokes mitochondrial membrane depolarization, and N-acetyl-l-cysteine is unable to reconstitute the physiological conditions. The present findings suggest that Bcl-2-like proteins play a main role in maintaining of the integrity of cellular components, e.g. mitochondria, rather than in controlling programmed cell death. Received 23 January 2001; received after revision 1 March 2001; accepted 1 March 2001     

烟草MnSoD cDNA基因的克隆及其在大肠杆菌中的表达

烟草锰超氧化物歧化酶(MnSOD)是一种由核基因编码并定位于线粒体的蛋白质,是植物体内清除超氧根阴离子的主要酶类．采用RT—PCR方法,从烟草(Nicotiana tubaccum)中克隆到MnSODeDNA基因(SodA),序列分析结果与文献报告的资料相比,核苷酸序列的同源性为99．9％,氨基酸序列的同源性为99．6％．该基因能在原核表达载体pBV221中正确表达,并表现出SOD酶活性．