FADS2基因在奶牛乳腺细胞中的过表达和干扰研究

为了研究脂肪酸脱氢酶2(fatty acid desaturases 2,FADS2)基因在奶牛乳腺细胞脂肪酸代谢中的作用,本研究在奶牛乳腺上皮细胞中对FADS2基因进行过表达和干扰,研究FADS2基因表达对脂肪酸合成相关基因的调控及对奶牛乳腺上皮细胞中甘油三酯含量的影响。针对FADS2基因的CDS序列设计siRNA和过表达载体pcDNA3.1-FADS2-EGFP,转染奶牛乳腺细胞检测FADS2基因过表达和干扰对脂肪酸代谢相关基因表达的影响及细胞中甘油三酯含量的变化。结果显示,试验成功获得过表达载体pcDNA3.1-FADS2-EGFP和干扰片段,转染细胞后具有良好的过表达和干扰效果。FADS2基因过表达后,1-酰基甘油磷酸酰基转移酶(AGPAT1)、固醇调节元件结合蛋白裂解激活蛋白(SCAP)、3-磷酸甘油转移酶(GPAM)、脂肪酸延长链5(ELOVL5)、乙酰辅酶A酰基转移酶1(ACAA1)、脂肪酸脱氢酶1(FADS1)、二酰基甘油转酰基酶1(DGAT1)和过氧化物酶体增殖激活受体α(PPARα)基因显著下调(P<0.05),脂滴蛋白2(PLIN2)基因极显著上调(P<0.01)。FADS2基因干扰过后可引起AGPAT1、GPAM、ELOVL5、ACAA1、PLIN2和FADS1基因显著上调(P<0.05),脂肪酸合成胰岛素诱导基因1(INSIG1)极显著上调(P<0.01),DGAT1和PPARα基因显著下调(P<0.05)。甘油三酯检测结果显示,FADS2基因过表达和干扰均可降低奶牛乳腺上皮细胞中甘油三酯的含量。综上所述,在奶牛乳腺上皮细胞中,FADS2基因能调控脂质合成相关基因的表达,对乳腺脂质合成具有调控作用。     

模拟降雨量变化与CO2浓度升高对羊草光合特性和生物量的影响

羊草作为欧亚大陆草原东缘干旱半干旱区域的优势种,现生产力已逐年降低。为明确未来降雨量变化和CO₂浓度升高对羊草草地变化的影响,我们模拟3个降雨量梯度(正常降雨量 40%)和两个CO₂浓度 (380±20,760±20 μmol/mol)进行了光合特性与生物量的研究。结果表明,降雨量变化和CO₂浓度的交互作用显著影响羊草的净光合速率等光合特性,地下生物量和根冠比,而不影响地上生物量。在目前CO₂浓度时随着降雨量的增加,以及在CO₂浓度升高时的降雨量由低降雨量增至正常降雨量,羊草的净光合速率、气孔导度、蒸腾速率和地上、地下及总生物量均显著增加,但在CO₂浓度升高,降雨量由正常降雨量增至高降雨量时上述指标无显著变化。根冠比在目前CO₂浓度时差异不显著,在CO₂浓度升高下低降雨量时显著增加。在降雨量相同条件下,CO₂浓度的升高使胞间CO₂浓度、水分利用效率显著增加,气孔导度和蒸腾速率显著降低。低降雨量时CO₂浓度升高对羊草净光合速率和生物量的促进作用显著高于正常降雨量和高降雨量,而高降雨量时CO₂浓度升高对羊草生物量没有促进作用。以上结果暗示着在目前CO₂浓度下,随着降雨量的增加,羊草生物量随之增加,在未来CO₂浓度升高的背景下,高降雨量年份对生物量的积累并无显著的促进作用,CO₂浓度升高可以补偿低水分条件对多年生根茎型禾草——羊草生长发育所造成的不利影响。     

大豆对CO2、O3污染的逆境反应

研究大豆对CO2、O3污染的逆境反应.以浓度分别700 mg/m^2的CO2、110 μg/m^3的O3以及二者混合气体,分别处理不同生育阶段的大豆叶片,测定其逆境下生理指标的变化.700 mg/m^2CO2条件下,大豆叶片中MDA、SOD、Oˉ2·ABA、CaM含量下降;可溶性糖（WSS）含量上升;110 μg/m^3O3浓度条件下,上述指标相反.CO2与O3混合,表现CO2具有抑制O3的氧化作用.伴随CO2浓度升高对O3浓度升高的胁迫产生同步抑制作用.     

黄芪多糖对大肠埃希菌侵入肠上皮细胞的保护研究

采用CaCo2细胞单层肠上皮系统感染模型,将5×10-5mmol/L,1×10-4mmol/L和2×10-4mmol/L 3种不同浓度的黄芪多糖及不含黄芪多糖DMEM细胞培养液分别加入已平铺于培养板的CaCo2细胞单层系统中与定量的大肠埃希菌混合培养,观察侵入细胞的大肠埃希菌量。结果黄芪多糖浓度为5×10-5,1×10-4mmol/L和2×10-4mmol/L,处理的CaCo2细胞液的细菌培养数量分别为252.5个±54.5个,240.8个±44.2个和292.7个±71.3个,而对照组为598.8个±114.8个,显示黄芪多糖能显著减少大肠埃希菌进入上皮细胞的数量,增强肠黏膜的屏障功能。     

Concordance between Results of Medium-term Liver Carcinogenesis Bioassays and Long-term Findings for Carcinogenic 2-Nitropropane and Non-carcinogenic1-Nitropropane in F344 Rats

This study was conducted to determine the concordance of results for a pair of structural isomers, 2-nitropropane (2-NP) and 1-nitropropane (1-NP), using the rat medium-term liver carcinogenesis bioassay (Ito test) and previously published long-term carcinogenicity tests. Male F344 rats were given a single intraperitoneal injection of DEN (200 mg/kg b.w.) to initiate hepatocarcinogenesis. After 2 weeks, they received per os 0, 0.8, 4 or 20 mg/kg/day of 2-NP or 1-NP six times a week and were subjected to two-thirds partial hepatectomy at week 3. Non-initiated groups receiving 0 or 20 mg/kg/day were also included. The animals were sacrificed for quantitative analysis of GST-P-positive foci at week 8. With the highest dose of 2-NP, significantly increased numbers and areas of GST-P-positive foci were demonstrated as compared with the respective control but were not noted with 1-NP. In the non-DEN-initiated groups, many small GST-P-positive foci of less than 0.2 mm in diameter were also induced in the rats treated with 2-NP at 20 mg/kg/day but were lacking with 1-NP. These results strongly support that 2-NP is a complete hepatocarcinogen with a potent initiation activity, whereas 1-NP is not.     

温性荒漠草原不同放牧强度草地锰和铜动态变化

为探讨不同放牧强度下温性荒漠草原土-草体系中微量元素锰和铜的变化,对土壤、牧草中锰和铜含量进行了测定。结果表明,除中牧区以外,其他放牧区土壤锰含量随季节的推移极显著下降(P0.01),夏季牧区极显著高于其他放牧区,冬春季和秋季则轻牧区极显著高于其他放牧区(P0.01)。除对照外,轻牧,中牧和重牧区土壤铜含量夏季极显著高于其他两季(P0.01),冬春季和秋季不放牧或轻牧区极显著高于中牧和重牧区(P0.01)。各放牧区牧草中锰和铜(除中牧区)含量均冬春季显著或极显著高于夏秋两季,同一季节不同放牧区则各有变化。     

Callose and β‐1,3‐glucanase inhibit Phytophthora cinnamomi in a resistant avocado rootstock

Phytophthora root rot (PRR) of avocado, caused by Phytophthora cinnamomi, is a significant threat to sustainable production wherever the crop is grown. Resistant rootstocks in combination with phosphite applications are the most effective options for managing this disease. Recently, the mechanisms underpinning PRR resistance have been investigated by the avocado community. Here, biochemical assays and confocal and scanning electron microscopy were used to investigate early defence responses in PRR resistant and ‐susceptible avocado rootstocks. Zoospore germination and subsequent hyphal growth for the pathogen were significantly inhibited on the surface of resistant avocado roots. When penetration occurred in the resistant R0.06 rootstock, callose was deposited in the epidermal cells, parenchyma and cortex of roots. In addition, β‐1,3‐glucanase was released early (6 h post‐inoculation, hpi) in response to the pathogen, followed by a significant increase in catalase by 24 hpi. In contrast, susceptible R0.12 roots responded only with the deposition of lignin and phenolic compounds incapable of impeding pathogen colonization. In this study, PRR resistance was attributed to a timely multilayered response to infection by P. cinnamomi.     

无角陶赛特羊与陶寒F2代羔羊生长发育对比简

为客观评价无角陶赛特羊产肉性能,及对小尾寒羊杂交改良的实际效果,于2012年7月～2012年11月对90日龄断奶纯种无角陶赛特羊和陶寒F2代羔羊（DHF2）的体重及体尺指标进行测定并进行差异性分析.结果表明：三月龄断奶陶寒F2代体重与纯种无角陶赛特羔羊差异不显著;陶赛特羊与陶寒F2代的公母羔体高、胸围均差异显著（P＜0.05）;公羔尻宽、胸深差异显著（P＜0.05）;母羔尻宽、胸深差异不显著（P＞0.05）;公羔体长差异不显著（P＞0.05）,母羔体长差异极显著（P＜0.01）;管围公母羔差异均不显著（P＞0.05）.可见从杂交育种到羔羊肉生产方面,杂交效果明显.     

Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

斜纹夜蛾化学感受蛋白SlitCSP2的表达模式定位及配体结合特性分析

研究斜纹夜蛾Spodoptera litura化学感受蛋白Slit CSP2的表达模式、组织定位及与配体结合特性,为揭示SlitCSP2在化学通讯中的功能提供理论依据。利用RT-qPCR检测了SlitCSP2在蛹及雌、雄成虫各组织的基因表达水平;Westernblot方法检测了SlitCSP2在幼虫期和雄成虫头部、触角和足部的蛋白表达水平;利用免疫组织化学技术对Slit CSP2在幼虫各龄期的分布情况进行定位分析;利用荧光竞争结合试验检测Slit CSP2与13种非挥发性化合物的结合特性。RT-qPCR结果表明,SlitCSP2在3龄幼虫表达量最高,3～6龄表达量逐渐降低;成虫阶段雄成虫足部及胸部和雌成虫头部及翅的表达量最高,且雄成虫头部、足部和胸部的表达量显著高于雌成虫。Western blot结果显示,SlitCSP2在3龄幼虫头部（去触角）的蛋白表达水平较高。免疫组化结果表明,Slit CSP2在幼虫阶段主要分布于头部、口器和胸足;在雄成虫中,Slit CSP2主要分布于足部前跗节和足部外骨骼内侧。荧光竞争结合试验表明,Slit CSP2与单宁（Ki=1.77）、花青素（Ki=2...