猪红细胞生成素受体基因突变与血常规性状关联分析

本研究旨在探索红细胞生成素受体基因(EPOR)影响红细胞相关性状的分子机理.对EPOR基因第1内含子和第4内含子突变位点g.705G＞T和g.2373C＞T,以飞行时间质谱进行个体基因型分型,在构建的大白猪×民猪F2代资源群体内开展其与6种血常规性状(红细胞压积、血红蛋白、平均红细胞血红蛋白量、平均红细胞体积、红细胞计数及红细胞分布宽度)的关联分析.结果表明,第1内含子突变位点g.705G＞T未发现与血常规性状显著关联;第4内含子突变位点g.2373C＞T群体内只发现2种基因型,其中CT基因型个体的红细胞压积显著高于CC基因型个体(P＜0.05),但并未发现与其他性状显著关联.结果显示,EPOR基因突变可显著影响红细胞压积,说明该基因可作为影响红细胞压积的主效候选基因开展深入研究.     

口蹄疫病毒Asial/JS/China/2005株基因组全长感染性克隆的构建

利用长距离RT-PCR技术扩增病毒基因组3'端覆盖口蹄疫病毒Asial/JS/China/2005株近全长的3个片段(共约7.5 kb),并利用单一酶切位点将其分别克隆到pBlueseriptSK+载体上.利用融合PCR扩增到基因组5'端含有15个C碱基的基因片段(约700 bp),并将其连接至pGEM-T载体.最后将这4个片段的阳性克隆装配至剔除T7启动子的低拷贝载体pcDNA3.1/Zeo(+)中构建该病毒株的全长cDNA克隆.以构建的FMDV Asial/JS/China/2005株全长cDNA为模板,使用TTRNA聚合酶在体外转录得到病毒RNA,通过脂质体将其导入BHK细胞获得拯救病毒.对收获的病毒分别用RT-PCR、间接免疫荧光、电子显微镜观察和乳鼠致病性分析结果证实,通过体外转录获得了具有感染性的口蹄疫病毒.该株感染性克隆的构建为深入研究口蹄疫病毒的致病机制及研制新型疫苗等奠定了基础.     

粉纹夜蛾(Trichoplusia ni)培养细胞对家蚕微孢子虫(Nosema bombycis)的吞噬过程观察

观察微孢子虫在非天然宿主细胞系的生物行为过程,有助于探究微孢子虫的侵染特异性机制。选用鳞翅目昆虫粉纹夜蛾(Trichoplusia ni,Tn)细胞系为材料,接种家蚕微孢子虫(Nosema bombycis,Nb)孢子后观察其吞噬过程。结果发现:经氢氧化钾发芽预处理的Nb孢子虽然可以在Tn培养细胞液中发芽,但未见发芽孢子的孢原质进入Tn培养细胞;在接种Nb孢子浓度为2.0×105mL-1以上时,观察到孢子通过内吞作用进入了Tn培养细胞,但未见孢子的增殖;随着时间的延长和进入Tn培养细胞的Nb孢子的增加,可引起Tn培养细胞的超微结构变化,并导致细胞的裂解。以上结果说明,虽未见Nb以发芽形式感染Tn培养细胞,但Nb能够被吞噬进入Tn培养细胞并破坏其结构。     

用卡片PCR法检测卡耶他环孢子虫

使用FTA卡片法提取卡耶他环孢子虫DNA,利用特异性引物组F1E/R2B、CC719/CRP999进行巢式PCR,得到1条298bp的特异条带。并以柔嫩艾美尔球虫、贝氏隐孢子虫等作为对照,结果显示,除卡耶他环孢子虫外,均无特异性条带。用卡耶他环孢子虫卵囊进行巢式PCR敏感性试验,结果显示敏感度最高达1.0×100个/mL。     

Application of PCR-HRM in the study of relationship between leptin gene promoter polymorphisms and liver cirrhosis

AIM: To analyze the feasibility of PCR-high-resolution melting curve analysis (HRM) for detecting the site mutations of C2549 and G2548 in leptin gene promoter from the patients with liver cirrhosis, and to explore the relevance between mutant genotypes and physiological and biochemical indexes in liver cirrhosis patients. METHODS: Compared with the method of PCR-restriction fragment length polymorphism (RFLP), the present research used the method of PCR-HRM to analyze the site mutations of C2549 and G2548 in leptin gene promoter in control group (n=100) and liver cirrhosis group (n=100). The physiological and biochemical indexes of the patients were also detected and compared. RESULTS: Leptin gene promoter polymorphism was detected using PCR-HRM with effectiveness, high flux and accuracy. Preliminary results showed that the main mutation of the patients with liver cirrhosis was in C2549 site, but not found in G2548 site. Leptin, free leptin index (FLI), fasting insulin (FINS) and insulin resistance index estimated by homeostatic model assessment (HOMA-IR) in liver cirrhosis group were higher than those in control group. Insulin sensitivity index (ISI) and soluble leptin receptor (sOB-R) in liver cirrhosis group were lower than those in control group with significant difference except leptin level. Meanwhile, FLI showed positively correlated with FINS and HOMA-IR (r=0.45, r=0.53, P<0.05), and negatively with ISI (r=-0.34, P<0.05). In the patients with liver cirrhosis, C2549A heterozygous mutation was predominant. The indexes of HOMI-IR, leptin, sOB-R and FLI of C2549A homozygotes and heterozygotes were higher than those of the wildtypes, which showed significant difference except leptin and sOB-R levels (P<0.05). CONCLUSION: PCR-HRM can be more accurate for identifying leptin promoter polymorphism. The increase in the frequency of C2549A mutation may be closely related with liver cirrhosis. Existence of hyperinsulinemia and insulin resistance may be correlated with leptin level in the patients with liver cirrhosis.     

山西菜田捕食性食蚜蝇优势种及其对菜蚜的控制效果

对山西菜田捕食性食蚜蝇优势种及对菜蚜的控制效果进行了研究，菜田常见捕食性食蚜蝇有黑带食蚜蝇，大灰食蚜蝇，斜斑鼓额食蚜蝇，野食蚜蝇，四条小食蚜蝇，短翅细腹食蚜蝇6种，以黑带食蚜蝇种群数量最大，平均占全年发生总量的36.3%，通过对食蚜蝇产卵量，卵化率，幼虫期捕食量和羽化率测试，筛选出的黑带食蚜蝇，大灰食蚜蝇，斜斑鼓额食蚜蝇幼虫对菜蚜均有较强的捕食能力，其中黑带食蚜蝇为山西菜田菜蚜捕食性食蚜蝇优势种，用黑带食蚜蝇三龄幼虫对菜蚜进行防治试验，结果以1：180防效最佳，72h后控制效果达93.5%。     

杏棉间作复合系统对棉花产量及纤维品质的影响

以大田棉花为对照(CK),研究了环塔里木盆地杏、棉间作系统对棉花的产量和纤维品质特性的影响.结果表明:杏、棉间作下棉花的产量及其构成因素相比对照均有不同程度的减小,且随着距杏树垂直距离的增加,其变化呈偏态分布趋势;在杏、棉间作区,棉花的比强度、伸长率和马克隆值明显减少,而纤维长度、整齐度、黄度和纺纱指数则形成了明显的增...     

Study on Developmental Expression of ApoCⅡ Gene mRNA in Liver Tissues of Pigs

The aim of this study was to investigate the developmental patterns of ApoCⅡ gene mRNA in liver in Mashen and Large White pigs, and study the relationship between the expression level of ApoCⅡ and the lipid metabolism in pigs.The mRNA relative expressions of ApoCⅡ gene in liver at seven stages of 1,30,60,90,120,150,and 180-day old in Mashen and Large White pigs were determined by quantitative Real-time PCR.The results showed that the developmental trend of ApoCⅡ mRNA expression in liver between Mashen and Large White pigs was different.The ApoCⅡ mRNA abundance was decreased from birth to 60-day old,then increased at 90-day old,and decreased again after that in Mashen pig.However,the relative expression amount in Large White pig was gradually decreased from birth to 150-day old and increased again at 180-day old.Except for the ApoCⅡ mRNA expression amount at 1-day old,the differences of the expression amount at other stages in Mashen and Large White pigs were significant or extremely significant (P<0.05 or P<0.01).The ApoCⅡ mRNA expression in liver was affected by age and breed,and could play an important role in lipid metabolism in pigs.     

基于分形视角的农村居民恩格尔系数变动趋势研究

恩格尔系数可以反映居民生活水平,也隐含着经济结构的信息。本文采用非线性定量分析工具——分形分析方法,通过各阶累计和将数据进行变换,建立变维分形模型,测算了1995～2008年我国农村居民恩格尔系数的变化趋势。研究表明,样本区间内农村居民恩格尔系数整体有降速放缓的趋势,前期降速显著,后期降速减缓;且依据模型测算显示未来5年内这种趋势将可能持续。最后,本文据此提出了简短的对策建议。     

虾夷马粪海胆早期生长发育的遗传力估计

以虾夷马粪海胆(Strongylocentrotus intermedius)为亲本，采用不平衡巢式设计方法和人工授精技术，每个雄性海胆配5个雌性海胆，每个雌性个体产生若干幼体，构成了11个父系半同胞家系和35个母系全同胞家系，分别测定了每个母系孵化后生长到3月龄和5月龄的全同胞幼海胆40-50个后代的体重和壳径，应用数量遗传学原理和半同胞组内相关分析法研究了虾夷马粪海胆早期生长发育性状的遗传力。结果表明，3月龄和5月龄的海胆体重的狭义遗传力估计值为0．339-0．523，壳径的狭义遗传力估计值为0．316-0．487。分析结果显示，雌性遗传方差组分均显著大于雄性遗传方差组分，雌性遗传方差组分存在显著的母性效应，表明由雄性遗传方差组分估计的遗传力准确可取，父系半同胞组内相关法计算的狭义遗传力是遗传力的无偏估计值。