Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens

We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.     

Effect of mutant β-catenin gene on the proliferation of hepatocytes

AIM:The β-catenin is a key molecule in the Wnt signal pathway, which plays a critical role in normal development and tumorigenesis. However, the mechanisms of the β-catenin on the cell growth control are still not completely defined. The aim of this study was to test the hypothesis that the mutant β-catenin may regulate the hepatocyte proliferation. METHODS: The immortalized murine hepatocyte cell line, AML12, was used for this study. A plasmid that contain mutant β-catenin S33Y was transfected into the AML12 cells and a stable cell line AML12S33Y was established. The cell growth property of this cell line and the parental cell were compared by flow cytometry analysis and direct cell count. The cells were also tested for the ability to form soft agar colonies, and the ability to form tumors in the severe immune deficient mice (SCID). RESULTS:1. The mutant β-catenin containing cell line AML12S33Y has higher proliferating index compared with the parental AML12 cells (P<0.01), suggesting that mutant β-catenin promotes cell growth. 2. The mutant β-catenin cells formed small colonies in soft agar after 4 weeks of culture, but did not generate tumor in SCID mice. CONCLUSION:The mutant β-catenin promotes liver cell growth.     

水稻瘤矮病毒P12蛋白抗血清制备及其应用

为了进一步研究水稻瘤矮病毒(Rice gall dwarf virus,RGDV)P12的功能,将RGDV-GX P12编码区亚克隆至原核表达载体,并导入大肠杆菌诱导表达;重组的P12融合蛋白经Ni-NTA His·Bind Resins纯化后用于免疫小鼠制备抗血清,并进行Western blotting分析。结果显示,约41 kD大小的RGDV P12融合蛋白可在大肠杆菌中高效表达;利用该重组蛋白所制备的抗血清可特异地与融合表达和非融合的P12蛋白发生强烈的免疫学反应。利用该特异性抗血清在病毒粒子样品中检测不到P12蛋白,而在病株总蛋白样品中可检测到1条与预期大小一致的明显条带,表明RGDV P12是一种非结构蛋白。     

农杆菌介导的hIL-12基因转化马铃薯的研究

构建CaMv35S启动子驱动人白细胞介素12基因(hIL-12)的植物表达载体pBI121-hIL-12,通过三亲杂交法将重组载体导入根癌农杆菌菌株EHA105,农杆菌介导法转化马铃薯茎段,经卡那霉素抗性筛选,获得28株抗性植株。通过PCR检测,22株为阳性,从PCR阳性植株中随机选取5株,ELISA法检测茎和叶中hIL-12的表达量,其中2株与对照组相比有显著性差异(p<0.001),hIL-12的表达量分别为(3 714.0±48.8)pg/g和(3 465.0±185.0)pg/g,初步表明外源基因hIL-12已经整合进马铃薯基因组中并得到了表达,为进一步研究重组hIL-12的分离纯化和生物学活性奠定了试验基础。     

勺夹式藠头排种器设计与试验

针对藠头（Allium chinense）种植劳动强度大、人工成本高等问题,该研究设计了一种由排种盘、取种勺、取投种凸轮、种箱、安装板等组成的勺夹式藠头排种器。对该排种器工作原理和取种、携种、投种过程进行理论分析,建立离散元仿真模型,对取种过程进行仿真分析,确定了最优种勺结构。以大叶藠为对象,以取种合格率和漏取率为试验指标,进行种勺直径、种勺深度、取种起始角、种面高度与种箱高度之比和取种速度5因素显著性筛选试验,并以种勺直径、取种起始角、种面高度与种箱高度之比为试验因素进行回归正交试验;采用Plackett-Burman试验法和Box-Behnken中心试验法建立取种合格率和漏取率的回归模型,并进行参数优化,获得最优参数组合为种勺直径27 mm、取种起始角30°和种面高度与种箱高度之比92.5%。在最优参数组合下进行台架试验,得到取种合格率91.17%,漏取率6.17%。田间试验结果表明在前进速度0.1～0.2 m/s的条件下,该排种器平均播种合格率、平均重播率和漏播率分别为63.10%、12.76%和24.14%。研究结果可为藠头播种机械的研究与设计提供参考。     

白桦BpMADS12基因启动子的克隆及表达分析

为探究白桦BpMADS12基因的功能,克隆了BpMADS12基因上游1750bp启动子序列,通过生物信息学对顺式作用元件进行分析,并利用农杆菌花序浸染法将其遗传转化入拟南芥,然后通过β-葡萄糖苷酸酶（GUS）组织化学染色检测BpMADS12启动子的组织表达特性及干旱胁迫应答。结果表明：BpMADS12启动子序列中含有与开花、激素及干旱响应等相关的顺式作用元件;该启动子在拟南芥中的表达模式呈现为在营养生长阶段不表达,而进入生殖生长阶段后,在根、花叶、花瓣、雄蕊、雌蕊及种子等各个组织部位中均表达;PEG胁迫后处理组拟南芥中GUS表达量低于未处理组。研究显示BpMADS12基因参与白桦的开花调节、激素应答、胁迫响应（干旱）等生物学过程,对生殖生长阶段各组织器官的发育有一定的调控作用,且负调控干旱响应途径。     

云线网蛾的生物学初步研究

云线网蛾Rhodoneura nitens是木兰科一些树木的食叶害虫,马褂木受害很普遍。此虫1a发生2代,以蛹越冬。越冬代和第1代成虫分别于4月下旬和7月下旬开始羽化。文章报道了其分布、寄主、生物学特性和天敌情况。     

维生素B12解冻液在绵羊人工授精中的应用

应用维生素B12注射液做绵羊冻精的解冻液,实施人工授精配种来提高绵羊受胎率和羔羊成活率。结果表明,试验组绵羊受胎率达96.0%,羔羊成活率达94.2%,而对照组绵羊受胎率为78.0%,羔羊成活率为85.4%,试验组与对照组绵羊受胎率和羔羊成活率均有显著性差异(P<0.05)。     

两种防霉剂与丙酸的组合对青贮牧草根霉菌毒力的研究

采用抑菌圈法研究了75%百菌清、50%多菌灵与99.5%丙酸在不同浓度下组合对高寒地区青贮饲草根霉菌的毒力。结果表明:丙酸(200mg/kg) 百菌清(400mg/kg)、丙酸(200mg/kg) 多酸灵(300mg/kg)、丙酸(100mg/kg) 多菌灵(300mg/kg)组合对根霉菌的毒力没有显著差异,但其毒力均极显著地高于其他处理;以丙酸(200mg/kg) 百菌清(400mg/kg)组合对青贮牧草根霉菌的抑菌作用最佳,其毒力分别是丙酸(200mg/kg) 多菌灵(300mg/kg)组合的1.05倍和丙酸(100mg/kg) 多菌灵(300mg/kg) 组合的1.20倍。     

Study on Antimicrobial and Antiviral Activities of Lysozyme From Marine Strain S-12-86 In Vitro

In this study, the in vitro antimicrobial and antiviral activities of the lysozyme from marine strain S-12-86 （LS） were investigated, The antimicrobial activity of LS was tested by minimum inhibition concentration （MIC） and minimum bactericidal concentration （MBC） method. The inhibiting effects of LS on pseudo rabies virus （PRV） in swine kidney cells （PK-15 ceils） were judged by cytopathogenic effect test （CPE）, The results showed LS had a broad antimicrobial spectrum against several standard strains including gram-positive bacteria, gram-negative bacteria, fungi, etc, The MIC of LS was 0.25-4.00 mg mL^-1 and its MBC was 0.25-8.00 mg mL^-1, respectively, Observation under the transmission electron microscope revealed that the cell wall of Candida albicans was distorted seriously, and the cytoplasm with many cavities was asymmetrical after being hydrolyzed by LS, The median cytotoxicity concentration （TC50） of LS was 100.0 μg mL^-1, the median effective concentration （EC50） was 0.46 μg mL^-1, and the selectivity index （TI = TC50/EC50） was 217. LS could inhibit PRV in PK-15 cells when it was added to cell culture medium at 0, 2, 4, 6, and 8 h after PK-15 cells had been infected by PRV. From the results, we concluded that LS had broad antimicrobial spectrum and good inhibiting effects on PRV,