Astaxanthin Normalizes Epigenetic Modifications of Bovine Somatic Cell Cloned Embryos and Decreases the Generation of Lipid Peroxidation

Astaxanthin is an extremely common antioxidant scavenging reactive oxygen species (ROS) and blocking lipid peroxidation. This study was conducted to investigate the effects of astaxanthin supplementation on oocyte maturation, and development of bovine somatic cell nuclear transfer (SCNT) embryos. Cumulus–oocyte complexes were cultured in maturation medium with astaxanthin (0, 0.5, 1.0, or 1.5 mg/l), respectively. We found that 0.5 mg/l astaxanthin supplementation significantly increased the proportion of oocyte maturation. Oocytes cultured in 0.5 mg/l astaxanthin supplementation were used to construct SCNT embryos and further cultured with 0, 0.5, 1.0 or 1.5 mg/l astaxanthin. The results showed that the supplementation of 0.5 mg/l astaxanthin significantly improved the proportions of cleavage and blastulation, as well as the total cell number in blastocysts compared with the control group, yet this influence was not concentration dependent. Chromosomal analyses revealed that more blastomeres showed a normal chromosomal complement in 0.5 mg/l astaxanthin treatment group, which was similar to that in IVF embryos. The methylation levels located on the exon 1 of the imprinted gene H19 and IGF2, pluripotent gene OCT4 were normalized, and global DNA methylation, H3K9 and H4K12 acetylation were also improved significantly, which was comparable to that in vitro fertilization (IVF) embryos. Moreover, we also found that astaxanthin supplementation significantly decreased the level of lipid peroxidation. Our findings showed that the supplementation of 0.5 mg/l astaxanthin to oocyte maturation medium and embryo culture medium improved oocyte maturation, SCNT embryo development, increased chromosomal stability and normalized the epigenetic modifications, as well as inhibited overproduction of lipid peroxidation.     

Cloning,Expression of Clostridium perfringens α-toxin Gene and Preparation of its Antisera

One pair of primers had been designed and synthesized based on the α-toxin gene of Clostridium perfringens.The complete α-toxin gene fragment was amplified by polymerase chain reaction (PCR), and then was cloned into pGEM-T Easy vector to construct pGEM-T-α.Digested with EcoRⅠ and Hind Ⅲ, a fragment of 1125 bp was cloned into the expression plasmid vector pET-28a(+).The recombinant plasmid was transformed into the BL21(DE3)plys and induced by 1.0 mmol/L IPTG at 37 ℃.The expression product was found to be 46.1 ku as expected one identified by SDS-PAGE, and confirmed by Western blotting with Clostridium perfringens type A antisera, indicating similar reactivity with native α-toxin.Recombinant α-toxin protein was simultaneously found in culture supernatant, postsonic supertanant and inclusion bodies, most protein was expressed in inclusion bodies, which indicated recombinant α-toxin protein was expressed in the extracellular, periplasm and cytoplasm.Recombinant α-toxin protein in postsonic supertanant could not make mice die, indicating its non-toxicity.Toxin-antitoxin neutralization test showed that antisera of recombinant α-toxin protein were specific to α-toxin.Upon immunization of rabbit with the recombinant α-toxin protein, antisera with high antibody titer neutralizing 100 MLD toxin per 1 mL were prepared.     

近53年青海省气候变化与粮食产量及气候生产潜力特征

利用青海省1961-2013年年平均气温、年降水量等资料,分析了青海省气候变化的特征,应用Thornthwaite Memorial 模型估算了青海省气候生产潜力,探讨了气候生产潜力与粮食产量的关系。分析结果表明,1961-2013年青海省年平均气温显著升高,年平均气温升温率达0.40 ℃10a-1;降水量也呈现增加趋势,增幅为6.0 mm10a-1;日照时数和相对湿度呈现减少趋势。自2002年以来的近10年期间,粮食总产量和气候生产潜力变化趋势有一定相关性且气候生产潜力从2002年开始有突变（r=0.29,P＜0.05）。近53年青海省气候生产潜力在323.39～478.48 gm-2a-1之间,年际间变化波动较大,呈现较弱的增加趋势,增加率为0.914 gm-2a-1。     

茉莉酸甲酯和磷酸氢二钾诱导柱花草对炭疽病的抗性

探究茉莉酸甲酯（Methyl Jasmonate,MeJA）和磷酸氢二钾（K2HPO4）诱导柱花草对炭疽病的抵抗作用。结果表明,二者浓度分别为0.001~0.1 mgmL-1和10~50 mmolL-1时,病情指数均显著降低（P0.05）,最佳诱导浓度分别是0.001 mgmL-1和30 mmolL-1,诱导效果分别是64.84 %和54.40 %。0.001 mgmL-1 MeJA处理组的过氧化物酶（POD）及多酚氧化酶（PPO）活性在48 h达到最大值,显著高于对照组。30 mmolL-1 K2HPO4处理组的POD与PPO活性分别在48和24 h达到最大值,显著高于对照组。两种诱抗剂处理的POD和PPO活性在接种早期增加了柱花草对炭疽菌的侵染的抵御能力,进而提高植株的抗病性。30 mmolL-1 K2HPO4处理组的过氧化氢酶（CAT）活性在48~72 h内上升,显著高于对照组,而0.001 mgmL-1 MeJA处理组的CAT活性除72 h显著低于对照组,其他时间点与对照组相差不大。说明在诱导柱花草抗病性方面,两种诱导抗性剂处理的诱抗效果与POD、PPO和CAT的酶活性的诱导的时间及程度有关。     

鸡、鹌鹑及其属间杂交种DMRT1b转录本初步分析

DMRT基因家族是一个与性别决定相关的发育基因家族。以鸡、鹌鹑及其属间杂交种胚胎为试验对象,分别采集孵化72 h的鸡、鹌鹑和杂交种胚胎,以DMRT1b为目的基因进行扩增,对DMRT1b基因原序列、扩增序列分析,应用Ex PASy网站对序列编码蛋白进行蛋白质生物信息学分析,结果发现杂交种DMRT1b基因较其父母本发生部分碱基缺失,预测蛋白质功能存在差异,推测由于基因缺失导致蛋白质不同,这种现象可能与杂交种早期死亡、不育存在相关性。研究结果为鸡与鹌鹑属间杂交种雌性致死与雄性不育的研究奠定基础。     

猪用加味玉屏风缓释微丸水提醇沉工艺研究

目的:考察猪用加味玉屏风缓释微丸的水提醇沉工艺。方法:以黄芪甲苷的含量及干膏得率作为评价指标,选用L9(34)进行正交试验。结果:猪用加味玉屏风缓释微丸最佳水提取工艺为药材分别加12倍量水,提取2次,提取时间分别为0.5 h;最佳醇沉工艺为浓缩药液相对密度1.15,加醇浓度70%,静置时间为12 h。结论:优选出的水提醇沉工艺科学、合理。     

河北省实行动物疫病风险监管的几点体会

实施动物疫病风险管理是控制动物疫病的一种有效方法,河北省对此开展了积极的探索,并取得显著成效。本文简要介绍了《河北省动物疫病风险评估与分级管理办法》和相关做法,探讨了实施动物风险管理在贯彻落实《动物防疫法》、实现动物疫病的科学监管、增强相对人的动物疫病风险管理意识、提高产业化水平、维护动物卫生监督执法人员的合法权益等方面的重要作用。     

猪霍乱沙门氏菌LAMP检测方法的建立与应用

利用猪霍乱沙门氏菌lacZ基因设计特异性引物,通过优化各种反应条件,建立了检测猪霍乱沙门氏菌的环介导等温扩增（LAMP）快速检测方法。此方法特异性好,灵敏度高,将细菌DNA稀释到10-8仍可检出,是PCR方法的1000倍。本研究建立的LAMP方法操作简便、灵敏度高、特异性强,为临床检测猪霍乱沙门氏菌和预防人猪霍乱沙门氏菌病提供了技术保障。     

基于翻转课堂模式的园林史教学改革探索

FC模式即"翻转课堂",也称为"颠倒课堂",是利用信息技术"翻转"知识传授和知识内化2个学习过程的全新教学模式。园林史是风景园林专业的重要专业基础课程,笔者经过历年园林史课程教学实践经验的积累,结合地区实际,探索了基于翻转课堂理念的园林史教学模式改革的实施途经,以期合理地解决好传统园林史教学革新中的相关问题。通过改进多媒体教学方式,高效利用课堂时间,充分发挥学生的学习主动性,进而提高教学效果和学生的学习兴趣,为园林史教学改革提供参考。