复方中药对内毒素引起的小鼠死亡率和多器官损伤的影响

目的：寻找有效的中药复方防治内毒素性多器官功能衰竭，降低内毒素血症小鼠的死亡率。方法：将四逆加人参汤、黄连解毒汤、参附青、参附青加丹参及清热解毒理气活血方药按常规方法煎制成药液(每毫升含1g生药)，以小鼠为实验对象，灌胃给药，3d后腹腔注射内毒素，观察小鼠生存率和肺、肝、肾、肠的病理变化。结果：与正常对照组(72h生存率为100％，n=20)比较，腹腔注射内毒素组小鼠72h生存率(42％，n=26)明显降低；四逆加人参汤、黄连解毒汤、参附青均不能提高内毒素血症小鼠生存率，而参附青加丹参反而降低内毒素血症小鼠生存率(72h生存率为8％，n=13)。清热解毒理气活血方药防治组小鼠72h生存率(68％，n=34)明显高于内毒索组(26％，n=38)，组织学观察显示肺、肝、肾、肠的病理变化明显减轻。结论：清热解毒理气活血方药能防治内毒素引起的多器官损伤，提高内毒素血症小鼠生存率。     

Overexpressing dominant negative MyD88 induces cardiac dysfunction in transgenic mice

Myeloid differentiation protein-88 (MyD88) is a crucial adaptor protein in the innate immune response. A protective role for MyD88 in normal cardiac function has been proposed in a surgical hypertrophic model. To assess the in vivo role of MyD88 in cardiac remodeling, we generated transgenic mice with cardiac-restricted expression of a dominant negative mutant of MyD88 (dnMyD88). Surprisingly, dnMyD88 transgenic mice displayed characteristic features of heart failure; including heart weight increase, cardiomyocytes enlargement, interstitial fibrosis, and re-expression of “fetal” genes. Echocardiographic examination of dnMyD88 hearts revealed dilated chamber volume and reduced cardiac contractility. DnMyD88 mice died from heart failure before they were 7 months old, as shown by Kaplan-Meier analysis. Additionally, the heart failure phenotype of dnMyD88 mice was associated with abnormal activation of the Akt/GSK-3β signaling pathway. These data provide the first evidence that normal MyD88 signaling is crucial for maintaining the physiological function of the adult heart.     

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     

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.     

红莲型水稻不育系与保持系线粒体DNA酶切分析

为研究红莲型水稻细胞质雄性不育的分子基础，我们提取纯化了红莲型水稻丛广４１Ａ和丛广４１Ｂ的线粒体ＤＮＡ，采用限制性内切酶酶切片段长度多态性（ＲＦＬＰ）方法比较了红莲型不育系和保持系线粒体ＤＮＡ酶切图谱，发现两者之间存在一定的差异，部分支持了细胞质雄性不育与线粒体ＤＮＡ有关的结论。     

小鼠口服猪卵透明带DNA避孕疫苗pVAX1-pZP3a的抗生育研究

目的：评价口服卵透明带DNA避孕疫苗pVAX1-pZP3a壳聚糖纳米微粒诱发的黏膜免疫反应、抗生育作用和对卵巢结构的影响，探讨用黏膜免疫避免卵透明带抗原免疫引起卵巢功能紊乱的可行性。方法：制备pVAX1-pzP3a壳聚糖纳米微粒，10只昆明系小鼠口服免疫，第3周和第6周分别加强免疫一次，剂量为O．5ml／只，含pVAXl-PZP3aDNA 50μg；对照组10只雌小鼠口服相同剂量不含pVAX1-pZP3a的壳聚糖。免疫前和免疫后隔周取小鼠血清、阴道冲洗液，用ELIASA法检测抗pzP3 IgA和IgG。首次免疫后第12周雌雄合笼交配，统计雌鼠怀孕情况；解剖取出双侧卵巢，制作石蜡切片，光镜观察卵巢病理学变化。结果：免疫组80％小鼠的血清和50％小鼠的阴道冲洗液可检测到抗pzP3 IgA，反应强度与持续时间有明显的个体差异；免疫组5只小鼠未怀孕，对照组全部怀孕，抗生育效果与合笼时抗pzP3 IgA反应有关；免疫组未怀孕小鼠卵巢组织结构与免疫组怀孕小鼠及对照组小鼠无差别。结论：口服pVAX1-pZP3a壳聚糖纳米微粒免疫小鼠能诱发生殖道黏膜免疫反应，发挥抗生育作用，对卵巢组织结构没有影响。黏膜免疫是克服卵透明带抗原免疫引起卵巢功能紊乱的一种值得深入研究的方法。     

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.     

Mitochondrial 16S rRNA sequence variations and phylogeny of the Chinese sisorid catfishes

Partial sequences of mitochondrial 16S rRNA gene were obtained by PCR amplification for comparisons among nine species of glyptosternoid fishes and six species of non-glyptosternoids representing 10 sisorid genera. There are compositional biases in the A-rich unpaired regions and G-rich paired regions. A-G transitions are primarily responsible for the Ts/Tv bias in unpaired regions. The overall substitution rate in unpaired regions is almost two times higher than that in the paired regions. Saturation plots at comparable levels of sequence divergence demonstrate no saturation effects. Phylogenetic analyses using both maximum likelihood and Bayesian methods support the monophyly of Sisoridae. Chinese sisorid catfishes are composed of two majorlineages, one represented by (Gagata (Bagarius, Glyptothorax)) and the other by “glyptosternoids Pseudecheneis“. The glyptosternoids may not be a monophyletic group. A previous hypothesis referring to Pseudecheneis as the sister group of monophyletic glyptosternoids, based on morphological evidence, is not supported by the molecular data. Pseudecheneis is shown to be a sister taxon of Glaridoglanis. Pareuchiloglanis might be paraphyletic with Pseudexostoma and Euchiloglanis. Our results also support the hypothesis that Pareuchiloglanis anteanalis might be considered as the synonyms of Pareuchiloglanis sinensis, and genus Euchiloglanis might have only one valid species, Euchiloglanis davidi.     

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。     

大壁虎线粒体12S rRNA基因片段的两种单倍型

通过测定大壁虎５个个体的线粒体１２Ｓ ｒＲＮＡ基因片段序列,发现大壁虎明显地分为两种单倍型。结合染色体组型上的微小差异,这样的种内差异是否可能达到亚种级水平,还有待于进一步研究。