基因枪介导红叶石楠遗传转化因素分析

 以红叶石楠（Photinia fraseri）的胚性愈伤组织为受体材料,利用基因枪法导入外源抗寒基 因rd29A,以期建立基因枪转化红叶石楠的技术体系。结果表明,基因枪转化红叶石楠的最佳组合条件为： 胚性愈伤组织在诱导培养基MS + 6-BA 2.0 mg · L-1 + NAA 5.0 mg · L-1 + 2,4-D 0.5 mg · L-1 + 琼脂5.0 g · L-1 + 蔗糖20 g · L-1 里,用0.3 mol · L-1 甘露醇处理15 ~ 16 h;基因枪轰击时添加12.5 mol · L-1 氯化钙 50 μL + 0.1 mol · L-1 亚精胺50 μL,每枪含金粉300 μg + 质粒DNA 3.0 μg,轰击距离9 cm,轰击压力1 100 psi, 轰击次数1 次。经多次筛选获得了6 株转基因植株,转化植株经PCR 检测和Southern 分析,证实了rd29A 已经整合到红叶石楠基因组中。     

不同苹果品种多酚含量的比较与提取利用研究

通过对不同苹果品种间多酚物质含量差异的比较分析,研究了影响物料粒度、料液比、pH值、提取温度、提取时间等因素对苹果多酚提取效率的影响,为充分开发利用苹果多酚提供工程学依据。     

滑子蘑多糖的抗肿瘤、抑菌活性及保湿特性

以滑子蘑菌丝体为试材,利用热水浸提法提取多糖,采用体外细胞试验、琼脂凹环法,检测了滑子蘑多糖的抗肿瘤、抑菌活性,并研究了多糖的保湿特性,以期为滑子蘑多糖的开发利用提供参考依据。结果表明:与空白试验组相比,滑子蘑多糖对小鼠巨噬细胞Ana-1的促增长率为48.65%,对胃癌细胞HGC-27的增殖抑制率为58.21%,对乳腺癌细胞BT-549的增殖抑制率为76.83%,对宫颈癌细胞Hela-229的增殖抑制率为63.57%;滑子蘑多糖对革兰氏阴性菌大肠杆菌、沙门氏菌及革兰氏阳性菌金黄色葡萄球菌均有抑制作用,且随着浓度的升高,抑制性增强;在30℃、RH 45%的恒温恒湿条件下,保湿性大小顺序为壳聚糖透明质酸山梨醇滑子蘑多糖甘油,在30℃、RH 80%的恒温恒湿条件下,保湿性大小顺序为透明质酸山梨醇壳聚糖滑子蘑多糖甘油,2种条件下滑子蘑多糖的保湿率都高于甘油,说明滑子蘑多糖有较好的保湿性。     

鸡柔嫩艾美尔球虫病抗性主成分分析评估模型的建立

为建立鸡活体可测球虫病抗性综合评估模型,探索鸡球虫病抗性选育新方法,以京海黄鸡为试验材料,根据国内外文献选择15个球虫抗性评价指标,通过比较鸡柔嫩艾美尔球虫感染组和非感染组抗性指标的差异,进一步筛选活体可测指标,并进行主成分分析,通过各主成分和综合主成分与所有抗性指标的相关分析,筛选最优抗性综合评估模型,并进行模型有效性筛选。在所选的14个活体可测抗性指标中,有6个指标在感染和非感染组间存在显著或极显著的差异(P0.05或P0.01),他们分别是感染后第3—5天体增重、感染后第8天血浆中CAT、GSH-Px、MDA、SOD和IFN-γ的浓度;主成分分析建立了6个单主成分和累计贡献率大于80%的综合主成分的抗性评估模型,各抗性评估模型计算的综合选择指数与15个抗性指标相关分析的结果表明,第一主成分模型和7个抗性指标相关极显著(P0.01),2个相关显著(P0.05),特别是与盲肠病变记分相关极显著(P0.01),其余6个主成分评估模型只与15个抗性指标中的2~3个相关显著或极显著(P0.05或P0.01),且与盲肠病变记分相关均不显著(P0.05);经检验第一主成分模型选择指数值的分布为正偏态分布,排序后其大小与盲肠病变表现一致。因此,第一主成分模型fi1=-0.64Zxi1+0.31Zxi2+0.80Zxi3-0.05Zxi4-0.08Zxi5+0.59Zxi6可以作为鸡柔嫩艾美尔球虫抗性选择的最佳综合选择指数用于鸡的抗病育种。     

闽南—台湾浅滩渔场中国枪乌贼生物学特性研究

头足类是最具开发潜力的渔业种群之一,然而其资源易受环境变化影响,复杂的交互作用致使二者空间关系呈现非均一性。中国枪乌贼 (Uroteuthis chinensis) 是南海北部近海重要的经济物种,且在渔业群落结构中占据优势种地位,故正确理解与掌握该物种资源-环境关系的空间特征,有助于该资源的保护与利用。基于南海北部近海2014年夏季的渔业资源调查数据,构建了地理权重回归模型 (Geographical weighted regression, GWR),探索该海域中国枪乌贼的资源分布与海洋环境关系的空间特征,阐述主要影响因子。模型评价指标结果表明,GWR的最小赤池信息准则 (Akaike Information Criterion, AIC) 和校正决定系数 (Adjusted R-Squared, Uroteuthis chinensis is an important economic species in the offshore waters of northern South China Sea, occuping a dominant position in the fishery community structure. Therefore, understanding the spatial characteristics of the resource-environment relationship is beneficial to its development, utilization and protection. Based on the fishery resources survey data in the offshore waters of northern South China Sea in summer of 2014, we established a geographically weighted regression (GWR) model to explore the spatial characteristics of the relationship between the resource distribution of U. chinensis and the marine environment in this area, and to reveal the main influencing factors. The results of model evaluation indexes show that the minimum Akaike information criterion (AIC) and adjusted R-Square (-6 mol/L for 24 h)-induced apoptosis of rat ventricular H9c2 myocytes, and to elucidate the possible mechanism. METHODS: The H9c2 cells were transfected with rAAV9-eGFP and rAAV9-eGFP-NF-κB p65-siRNA at multiplicity of infection (MOI)=4×10⁶ vg/cell. eGFP expression in the cells was observed under an inverted fluorescence microscope, and the percentage of eGFP positive cells was determined by flow cytometry. The expression of p65 was determined by Western blot. CCK-8 assay was used to measured the viability of transfected H9c2 cells. The apoptosis of the cells transfected with the virus and with Ang Ⅱ stimulation was analyzed by flow cytometry. RESULTS: The cells began to exhibit eGFP expression on the 2nd day after transfection. The fluorescence intensity was increased over the time of transfection. eGFP expression reached the maximum on the 5th day, and the transfection efficiency was (52.7±1.9)% at this time point. Compared with blank control group, no significant effect of AAV9 on the viability of H9c2 cells was observed. In resting state, p65 in the H9c2 cells had a certain activity. After Ang Ⅱ stimulation, the activity of p65 was obviously increased, while transfection of rAAV9-eGFP-NF-κB p65-siRNA effectively inhibited the expression of p65. The apoptosis of H9c2 cells in Ang Ⅱ stimulation group was significantly higher than that in blank control group, while transfection of rAAV9-eGFP-NF-κB p65-siRNA effectively inhibited apoptosis of H9c2 cells. CONCLUSION: Transfection of rAAV9-eGFP-NF-κB p65-siRNA effectively inhibits the expression of p65 gene of NF-κB pathway in the H9c2 cells without causing cell growth inhibition, and reduces the apoptosis induced by Ang Ⅱ.     

肝毒净颗粒的质量控制研究

对栀子和黄芩进行薄层色谱鉴别,采用高效液相色谱法测定肝毒净颗粒中绿原酸含量.色谱柱为C18柱,流动相为乙腈-0.4%磷酸水溶液（8∶ 92）,检测波长为324.5 nm.绿原酸在0.161 6～1.292 8 mg范围内,峰面积与其呈良好线性关系（r= 0.999 9）,平均回收率为99.19% （n=6）,RSD=1.26%.本方法可用于控制肝毒净颗粒的质量.     

新城疫病毒ClassⅠ强毒株磷蛋白在细胞中的表达与定位

根据Class Ⅰ新城疫病毒分离株9a5b编码序列设计特异性引物扩增P蛋白基因,原核表达后利用纯化的蛋白免疫BALB/c小鼠制备单克隆抗体,经二次亚克隆后筛选到4株针对P蛋白的单克隆抗体,利用所得单抗进行P蛋白在细胞内定位分析,NDV 9a5b株按5 M.O.I感染量接种DF1单层细胞,当PI=4 h时,P蛋白呈点状弥散分布于细胞浆内;PI=6~8 h时,P蛋白聚集于细胞核周围;PI=12 h时,P蛋白呈单极化聚集于细胞核一侧;而PI=16 h后开始形成合胞体,P蛋白在单个细胞中仍呈现单极分布;当PI=48 h时,细胞核已发生碎裂,荧光强度有所减弱。本研究为深入开展P蛋白在病毒增殖以及细胞周期中的作用机制研究奠定了基础。