PI3K/Akt信号通路相关的生物学调控机制研究进展

PI3K/Akt信号通路是由酶联受体介导的信号转导通路,该通路不仅参与多种生长因子、细胞因子和细胞外基质等的信号转导,同时还参与细胞增殖、分化、凋亡和葡萄糖转运等多种细胞功能的调节,特别是在细胞凋亡、细胞存活以及调控细胞糖代谢等方面具有重要作用。本研究综述了PI3K-Akt信号通路的结构组成、通路活化、通信过程、调控机制及其生物学功能等方面的研究进展,为进一步研究PI3K/Akt信号通路的生物学调控作用机制提供启示。     

一次陆源降雨污水引起血红哈卡藻赤潮的成因

以2007年烟台四十里湾海域血红哈卡藻(Akashiwo sanguinea Hirasaka)赤潮为对象,研究赤潮消长与水环境因子的关系。研究发现大量陆源降雨污水输入后,海水盐度急剧下降、营养盐大幅增加,特别是活性磷酸盐浓度明显增加,促进了血红哈卡藻的生长繁殖并最终形成赤潮。赤潮发生前海区第一优势种为尖刺拟菱形藻(Pseudo-nitzschia pungens Halse),优势度0.47(0.42—0.52),多样性指数2.63(2.43—2.89);赤潮发生时血红哈卡藻密度范围1.05×105—4.10×106个/L,优势度0.92(0.83—0.99),多样性指数0.27(0.15—0.64);赤潮消退后中肋骨条藻(Skeletonema costatum Cleve)为第一优势种,优势度0.49(0.43—0.55),多样性指数2.46(2.19—2.84)。赤潮的发生、发展、消亡与化学需氧量(COD)、无机氮(DIN)、活性磷酸盐(DIP)、富营养化指数(E)呈显著正相关(P<0.05),与盐度呈显著负相关(P<0.01)。赤潮前、后该海域为贫营养、P限制、叶绿素a含量中等,赤潮期间该海域为富营养、P限制、叶绿素a含量高。通过影响力评定,活性磷酸盐、COD、盐度是此次赤潮发生的主要诱发因子,当活性磷酸盐含量低于0.3μmol/L时,硅藻逐渐取代甲藻,此次赤潮消散。     

An atlas of CNV maps in cattle,goat and sheep

Copy number variation(CNV) is the most prevalent type of genetic structural variation that has been recognized as an important source of phenotypic variation in humans, animals and plants. However, the mechanisms underlying the evolution of CNVs and their function in natural or artificial selection remain unknown. Here, we generated CNV region(CNVR) datasets which were diverged or shared among cattle, goat, and sheep, including 886 individuals from 171 diverse populations. Using 9 environmental factors for genome-wide association study(GWAS), we identified a series of candidate CNVRs, including genes relating to immunity, tick resistance, multi-drug resistance, and muscle development. The number of CNVRs shared between species is significantly higher than expected(P0.00001), and these CNVRs may be more persist than the single nucleotide polymorphisms(SNPs) shared between species. We also identified genomic regions under long-term balancing selection and uncovered the potential diversity of the selected CNVRs close to the important functional genes. This study provides the evidence that balancing selection might be more common in mammals than previously considered, and might play an important role in the daily activities of these ruminant species.     

Cell death-inducing DFFA-like effector c (CIDEC/Fsp27) gene: molecular cloning, sequence characterization, tissue distribution and polymorphisms in Chinese cattles

Cell death-inducing DFFA-like effector c (CIDEC) protein, also known as fat specific protein 27 (Fsp27), is localized to lipid droplets. CIDEC protein is required for unilocular lipid droplet formation and optimal energy storage in addition to controlling lipid metabolism in adipocytes and hepatocytes. Research found that Ad-36 could induce lipid droplets in the cultured skeletal muscle cells and this process may be mediated by promoting CIDEC expression. The content of intermuscular fat is an important index for evaluation of beef quality, so the CIDEC gene appeared to be a candidate gene for regulation of intermuscular fat, however similar research for the bovine CIDEC gene is lacking. This paper examined the tissue expression profile of CIDEC gene in cattle using real-time RT-PCR to suggest that bovine CIDEC is highly expressed in adipose tissue. In addition, the Bovine CIDEC gene was cloned and inserted into the eukaryotic expression vector pET-28a(+), whereupon recombinant bovine CIDEC protein was induced and identified by Western-blot. A phylogenetic analysis showed that the animo acid sequence of bovine CIDEC was closer to mammalian CIDEC than rasorial CIDEC. We found ten single nucleotide polymorphisms sites (SNPs) in bovine CIDEC gene, of which SNP 2, 3, 4, 6 and 9, and SNP 8 and 10 were in complete linkage disequilibrium, respectively. SNP 1, 2 and 10 were used in further haplotype studies. Eight different haplotypes were identified in 973 cattle, of which haplotype 8 predominated with frequencies ranging from 42.90 to 54.30 %. This research provides a basis for future functional studies of CIDEC in cattle.     

莱州湾金城海域浮游动物群落结构及与环境因子的关系

于2009年和2010年3—12月在莱州湾金城海域对浮游动物群落结构及其环境因子进行了连续20个月份的综合调查,共记录到浮游动物75种,分属14个不同大类。浮游动物以桡足类和浮游幼虫居多,分别鉴定出22种、20类,占种类总数的29.3%、26.7%,其次为水螅水母类,鉴定出11种,端足类、被囊动物、腹足类和栉水母类分别鉴定出6种、4种、4种、2种,其它枝角类、毛颚动物、等足类、糠虾类、磷虾类以及钵水母类各鉴定出1种。主要优势种为强壮箭虫(优势度Y=0.24)、中华哲水蚤(Y=0.13)、墨氏胸刺水蚤(Y=0.07)、以及洪氏纺锤水蚤(Y=0.02)。2009年与2010年浮游动物群落结构的变化趋势比较稳定,同一季节群落结构相似度达到40%以上。浮游动物丰度分别在2009年及2010年的5月份达到最高值951.65个/m3、1348.14个/m3。种类数分别在2009年9月和2010年6月达到最高值48种、40种。多样性指数分别在2009年10月及2010年5月达到最高值2.6、2.7,呈现明显的季节变化特征。典范对应分析(Canonical correspondence analysis,CCA)显示,水温和盐度是影响浮游动物群落结构最主要的影响因子,水温与浮游动物种类数的相关系数达到0.87(P0.01)。盐度与浮游动物数量的相关系数为0.484(P0.05)。透明度、化学耗氧量、pH、叶绿素及总氮总磷等环境参数对浮游动物群落结构的影响较弱。     

Identification of bovine NPC1 gene cSNPs and their effects on body size traits of Qinchuan cattle

NPC1 gene is an important gene closely related to the Niemann–Pick type C (NPC). Mutations in the NPC1 gene tend to cause Niemann–Pick type C, a lysosomal storage disorder. Previous studies have shown that NPC1 protein plays an important role in subcellular lipid transport, homeostasis, platelet function and formation, which are basic metabolic activities in the process of development. In this study, to explore the association between the NPC1 gene variation and body size traits in Qinchuan cattle, we detected four novel coding single nucleotide polymorphisms (cSNPs) in the bovine NPC1 gene, including one missense mutation (SNP1) and three synonymous mutations (SNP2, SNP3 and SNP4). Population genetic analyses of 518 individuals and association correlations between cSNPs and bovine body size traits were conducted in this research. A missense mutation at SNP1 locus was found to be significantly related to the heart girth, hip width and body weight (P < 0.01 or P < 0.05, 3.5-year-old). Two synonymous mutations at SNP2 and SNP3 loci also showed significant effects on hip width (P < 0.05, 3.5-year-old). One synonymous mutation at SNP4 locus showed significant effect on body weight (P < 0.05, 2.0-year-old). Combined haplotypes H₂H₆ and H₆H₆ showed significant effects on body size traits such as heart girth, hip width, and body weight (3.5-year-old, P < 0.01 or P < 0.05). This study provides evidence that the NPC1 gene might be involved in the regulation of bovine growth and body development, and may be considered as a candidate gene for marker assisted selection (MAS) in beef cattle breeding industry.     

鸭病毒性肝炎二价灭活疫苗 (DHAV-SH和DHAV-FS株) 的制备和评价

我国鸭甲型肝炎病毒(DHAV)的快速变异及广泛流行给养鸭业造成了较大的经济损失,为研究鸭病毒性肝炎二价灭活疫苗(DHAV-SH和DHAV-FS株)的制备和评价方法,首先对我国多个鸭场进行血清型流行病学分析,从201株DHAV-1、38株DHAV-3中筛选出6株毒力较强的DHAV,验证了DHAV-1和DHAV-3为我国流行优势株,并对6株DHAV进行ELD50和LD50测定,筛选疫苗候选株后经传代确定疫苗毒种,选取F5代鸭胚胚体研磨液作为疫苗毒种,经甲醛灭活后制成3批水包油包水(W/O/W)型乳剂二价灭活疫苗实验室制品。通过安全性试验、抗体中和试验、攻毒保护及免疫交叉保护试验发现:疫苗安全性良好,雏鸭免疫后第7天可以检测到DHAV中和抗体,第14-21天的攻毒保护率为90%-100%,免疫持续期可达5周以上,DHAV-SH和DHAV-FS之间的交叉保护为20%-30%。研究表明本试验研制的鸭病毒性肝炎二价灭活疫苗实验室制品安全且高效,为我国DHAV预防和控制提供了一种新方法和新产品。     

Application of mathematical expectation (ME) strategy for detecting low frequency mutations: An example for evaluating 14-bp insertion/deletion (indel) within the bovine PRNP gene

The detection method based on the mathematical expectation (ME) strategy is fast and accuracy for low frequency mutation screening in large samples. Previous studies have found that the 14-bp insertion/deletion (indel) variants of the 3′ untranslated region (3′ UTR) within bovine PRNP gene have been characterized with low frequency (≤5%) in global breeds outside China, which has not been determined in Chinese cattle breeds yet. Therefore, this study aimed to identify the 14-bp indel within PRNP gene in 5 major Chinese indigenous cattle breeds and to evaluate its associations with phenotypic traits. It was the first time to use ME strategy to detect low frequency indel polymorphisms and found that minor allele frequency was 0.038 (Qinchuan), 0.033 (Xianan), 0.013 (Nanyang), 0.003 (Jiaxian), and zero (Ji'an), respectively. Compared to the traditional detection method by which the sample was screened one by one, the reaction time by using the ME method was decreased 62.5%, 64.9%, 77.6%, 88.9% and 66.4%, respectively. In addition, the 14-bp indel was significantly associated with the growth traits in 2 cattle breeds, with the body length of Qinchuan cattle as well as the body weight and waistline of Xianan cattle. Our results have uncovered that the method based on ME strategy is rapid, reliable, and cost-effective for detecting the low frequency mutation as well as our findings provide a potential valuable theoretical basis for the marker-assisted selection (MAS) in beef cattle.     

莱州湾金城海域网采浮游植物年际变化及与环境因子的关系

为了解莱州湾金城海域浮游植物群落年际变化特征,在该海域设置了15个监测站位,于2009年和2010年采用浅水Ⅲ型浮游生物网逐月连续进行24个航次浮游植物调查,同步监测其他环境因子。浮游植物样品经鲁格氏溶液固定后在OLYMPUS BX_(51)显微镜下进行分类、鉴定、计数,共鉴定出浮游植物81种,隶属于硅藻门、甲藻门、黄藻门和金藻门4门25科43属。其中硅藻16科33属67种,占种类数的82.7%;甲藻6科7属11种,占种类数的13.6%;黄藻2科2属2种,占种类数的2.5%;金藻1科1属1种,占种类数的1.2%。2009年和2010年金城海域浮游植物种类组成和比例基本一致,浮游植物丰度年际变化和种类数年际变化均呈现明显的双峰模型,每年2月和9月为高峰期,5月为全年最低值。2009年和2010年细胞丰度均值分别为465×10~4(38.4×10~4—1351×10~4)个/m~3和457×10~4(41.8×10~4—1380×10~4)个/m~3,均低于1982—1983年平均水平。浮游植物群落年际相似性在60%水平聚类分为四个类群,其中Ⅰ类为5月份样品,优势种为夜光藻和洛氏角毛藻;Ⅱ类为11—4月份样品,优势种有中肋骨条藻、旋链角毛藻和夜光藻;Ⅲ类为8、9、10月份样品,优势种为洛氏角毛藻和丹麦细柱藻;Ⅳ类为6、7月份样品,优势种为洛氏角毛藻和尖刺拟菱形藻。浮游植物群落Shannon-Winner多样性指数平均2.715 (1.446—3.807),Margalef丰富度指数平均2.418 (1.545—3.153),Pielou均匀度指数平均0.523 (0.263—0.706)。2009年和2010年多样性指数和均匀度高峰值出现在3月和10月,低值出现在7月和9月;均匀度指数年际变化低值出现在6—9月。浮游植物群落结构主要受水温和盐度环境因素的影响,其中水温与洛氏角毛藻丰度呈极显著正相关(R=0.613,P0.01),与旋链角毛藻、中肋骨条藻和夜光藻丰度呈极显著负相关(R=-0.662,-0.649,-0.649,P0.01);化学需氧量与旋链角毛藻和中肋骨条藻均呈显著负相关(R=-0.589,-0.480,P0.05);盐度与中肋骨条藻呈极显著负相关(R=-0.585,P0.01);其他环境因子与各优势种丰度相关性不明显。