两广地区2011--2012年H9N2亚型禽流感病毒的HA基因进化分析

为探究两广地区H9N2亚型禽流感病毒（avianinfluenzavirus,AIV）的变异情况及分子流行规律,于2011-2012年从该地区发病鸡群中共分离到16株H9N2亚型A1V,并对分离株HA基因进行测序与进化分析。结果表明,分离株HA基因开放阅读框全长均为1683bp,编码560个氨基酸;HA基因核苷酸同源性为88．7％～99．6％,编码氨基酸同源性为91．8％～99．5％。本试验分离毒株与国内疫苗株（GD-SS、SH—F和SD-6）的核苷酸同源性在90．1％～92．6％之间,推导的氨基酸序列同源性在91．6％～94．8％之间。进化分析显示分离株可分为Group1和Group2两个亚分支,与疫苗株均属于欧亚谱系的Y280分支,但亲缘关系较远。分离株HA蛋白裂解位点附近序列有3种形式：PARSSR＋GLF、PSRSSR＋GLF和PARLSR0GLF,均无连续碱性氨基酸的插A,符合低致病性AIv的特征。本试验发现分离株GD4、GX2在HA1的127、295位分别增加一个潜在的糖基化位点;除分离株GD5和GD6外,其余分离株在HAl的216位发生Q216L氨基酸突变,表明其存在感染人的可能性。     

昆虫黑化反应的分子机制研究进展

目的克氏原螯虾(Procambarus clarkii)属于节肢动物门甲壳纲十足目,也称淡水小龙虾。多巴脱羧酶(dopa decarboxylase)作为小龙虾先天免疫应答中的一种关键酶,在抵御外来病原体的侵害上起到了重要作用。本研究以克氏原螯虾为材料,克隆了Pc-ddc cDNA序列并对其进行生物信息学及组织mRNA表达分析。方法通过分子生物学技术构建大肠杆菌表达载体,并进行生物信息学分析,同时应用qRT-PCR技术分析其mRNA组织表达分布。结果生物信息学分析表明：获得了开放阅读框全长为1 425 bp的Pc-ddc基因的cDNA序列,编码474个氨基酸残基,多肽的理论分子量为52.99 ku,蛋白分子式为C₂₃₈₄H₃₆₈₉N₆₄₇O₆₇₄S₂₅,等电点为5.98,为弱酸性蛋白且N-末端无信号肽。亚细胞定位表明：Pc-DDC蛋白在线粒体中最多;组织表达分析表明：Pc-ddc mRNA在检测的6个组织中差异表达,其中,淋巴组织表达量最高。结论本研究为克氏原螯虾等无脊椎动物先天免疫系统的深入探索提供了理论支撑。     

云南某农村猪戊型肝炎病毒分子流行病学调查

目的了解昆明市某猪场戊型肝炎病毒感染情况,进一步探究病毒基因型及分子进化情况,为戊型肝炎防控提供参考。方法从该猪场随机采集39份新鲜粪便,用逆转录巢式聚合酶链反应对HEV ORF2部分基因片段扩增、克隆及测序;应用生物信息学软件进行同源性分析并构建系统进化树。结果PCR检测结果发现：该猪场核酸阳性率达35.9% (14/39)。同源性分析结果表明：14株流行毒株核苷酸序列与GenBank中HEV I~IV 型参考株的同源性分别为77.4%~83.9%、81.0%~83.2%、78.8%~86.9%和90.5%~98.5%,全部流行株均属基因IV型。进化分析结果表明：YN-KM-9与该猪场其他流行毒株同源性较远,应属不同基因亚型。结论该猪群HEV IV感染率较高,对公共卫生安全构成潜在威胁。     

Effects of vegetable oil supplementation on feed intake,rumen fermentation,growth performance,and carcass characteristic of growing swamp buffaloes

Fifteen, one year old swamp buffalo males with average liveweight of 200.5 ± 9.5 kg were randomly assigned according to a completely randomized design to receive three dietary treatments of supplemental vegetable oils in concentrate contained 140, 750, and 16 g of crude protein, total digestible nutrient, and ether extract, respectively (T1 = unsupplemented, T2 = supplemental coconut oil and sunflower oil in ratio 50:50 at 6% of concentrate, and T3 = supplemental sunflower oil at 6% of concentrate) during a 5-month feeding trial. Urea-treated rice straw was offered ad libitum and concentrate was supplemented at 150 g/kg BW. The results revealed that supplementation of vegetable oils decreased dry matter intake in sunflower oil supplemented group. Combination of coconut oil and sunflower oil supplementation resulted in similar average daily gain as those found in control and in sunflower oil supplemented groups. Rumen microorganism population, ammonia–nitrogen, blood urea–nitrogen and total volatile fatty acid concentrations were decreased as vegetable oils were supplemented. Oil supplementation slightly reduced growth rate, hot carcass percentage, loin eye area particularly on sunflower oil supplemented group. Based on these findings, vegetable oil resulted in decreased performance of swamp buffaloes although the combination of coconut oil and sunflower oil could reduce negative effect of unsaturated oil supplementation. However, further analyses of conjugated fatty acids (CLA) in meat could offer useful information in determining pronounced effects of vegetable oil supplementation.     

固定效应对槟榔江水牛体重影响的研究

[目的]槟榔江水牛为我国目前为止发现的唯一本土河流型水牛,具有产奶性能高、肉用性能好的特点,为了进一步阐明槟榔江水牛的生长发育规律,本文收集云南腾冲县巴福乐槟榔江水牛2004~2011年生长发育数据资料,研究性别、月份和年度等固定效应对体重的影响。[方法]应用SAS分析性别、月份、年度等非遗传因素对槟榔江水牛体重的影响,并且用MTDFREML软件分析了固定效应值。[结果]本研究结果表明,槟榔江水牛性别对初生重具有显著影响,公犊平均初生重(35.04kg)显著高于母犊(32.63kg);月份对母牛初生重没有显著影响,公牛的效应值比母牛的较大。[结论]月份对槟榔江水牛的体重有显著的影响,可能与饲料的季节性供给有关。     

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.     

水稻矮缩病毒基因组及毒粒三维结构的研究进展

水稻矮缩病毒(Rice dwarf virus,RDV)为双层壳、二十面体的双链RNA病毒,是水稻的重要病原物之一.文中比较了RDV中国福建分离物与日本分离物基因组各片段核苷酸序列的相似性,发现RDV两个不同分离物基因组相应片段的相似性均在92%以上,最高可达96%;分析了相应基因组片段编码的结构蛋白P1、P2、P3、P5、P7、P8及以前一直被认为是非结构蛋白的P9和非结构蛋白Pns4、Pns6、Pns10 、Pns11和Pns12在RDV复制、组装过程中的功能;回顾了RDV粒子三维结构的研究概况,对RDV粒子内、外层衣壳的晶体学结构作了较详细的描述;简要介绍了RDV复制与组装的机制,发现核心蛋白与dsRNA间相互作用成为一个单元是病毒RNA的分拣及包装到病毒粒子核心内的可能机制.同时,指出基于PDR的植物抗病毒基因工程在水稻矮缩病毒防治上的可能性.     

美洲黑杨基因资源收存及其遗传评价的研究

从１７个国家引进３３１个黑杨派无性系（其中５２个美洲黑杨无性系）,在山东省长清县营建我国第一个黑杨派无性系基因库。连续１０年对基因库内美洲黑杨无性系进行了多性状系统研究,结果表明各无性系间在物候期、生长、生根、抗寒和抗病虫等方面均存在着显著的差异,遗传变异丰富;系间的主要性状与无性系起源纬度及各性状间存在一定相关性。主要材料性状变异的研究结果表明,３８个８年生美洲黑杨无性系在木材基本密度、纤维长度无性系间变异达到极显著水平;木材密度和纤维长度与树高和胸径呈一定相关性;木材密度和纤维长度这两个性状在遗传上相互独立,受不同遗传机制控制。利用分子遗传标记ＲＡＰＤ技术研究库内美洲黑杨无性系ＤＮＡ多态性,结果表明本库美洲黑杨ＤＮＡ多态率为８６％,再次证明本库美洲黑杨遗传多样性高,可为我国杨树改良提供丰富的育种材料。     

栽培柳枝稷不同生长时期对伴生杂草生物量及密度的影响

以陕西定边县引种的8个柳枝稷(Panicum virgatum L.)品种(Blackwell,Cave-in-Rock,Dakota,Forest-berg,Illinois USA,Nebraska 28,Pathfinder和Sunburst)及行间杂草作为研究对象,通过计算柳枝稷对杂草的抑制率(RI),分析其生长指标与杂草生长指标的相关关系,判定不同柳枝稷品种在不同生育期对杂草生长的抑制效应,为其引种及田间杂草管理提供依据。结果表明:在生长初期(6月30日),Illinois USA杂草抑制率仅为3.4%,Cave-in-Rock和Forestberg对杂草生长抑制率分别为86.0%和74.9%;经过2次人工除草后(8月30日),Cave-in-Rock和Forestberg的杂草抑制率分别为85.7%和95.7%;生长后期(10月13日),仅Cave-in-Rock对杂草生长有88.5%的抑制率。回归分析显示,生长中后期(8月30日),柳枝稷地上部及根系生物量显著影响对杂草的抑制率(R2=0.31,0.54;P<0.05)。Pearson相关分析发现,Cave-in-Rock的株高、地上及根系生物量均显著降低杂草单株生物量(r=-0.97,-0.99,-0.99;P<0.05);Forestberg的植高与杂草密度显著负相关(r=-0.97,P<0.05)。     

犬尿结石的化学成分分析研究

应用物理学特征观察、化学定性分析、X射线衍射分析等方法,分析了临床收集的1份犬肾结石样品、2份犬膀胱结石样品的化学成分,并根据结石的形成原因、机理探讨其防治方法。结果表明:肾结石样品为混合型结石,主要成分为水合磷酸氢镁、水合磷酸铁及水合磷酸钙,另外还可能含有少量水合硫酸钙;2份膀胱结石样品主要成分均为六水磷酸铵镁。对尿结石的成分分析,有助于了解结石形成机制,为尿结石的防治奠定一定基础。