始丰微型牛体型与外周血cAMP、PTH和TGF-β水平关联性分析

为阐明始丰微型牛的生长发育相关的生化特性,采用ELISA方法对15月龄的始丰微型牛和作对照的正常体型天台黄牛测定并比较分析外周血中三个生长发育相关因子:cAMP、PTH与TGF-β.结果表明:微型牛组TGF-β含量(192.6 ng/L)极显著低于对照组(229.5 ng/L)(P＜0.01),cAMP和PTH含量与对...     

Interleukin-1β inhibits AKT to down-regulate eNOS Ser1177 phosphorylation in human umbilical vein endothelial cells

AIM To investigate whether interleukin-1β （IL-1β） regulates endothelial nitric oxide synthase （eNOS） phosphorylation at Ser1177 site in human umbilical vein endothelial cells （HUVECs）, and to explore its possible mechanism. METHODS The HUVECs were randomly divided into normal control group, tumor necrosis factor-α （TNF-α） group, IL-1β group, IL-6 group, SC79 ［protein kinase B （PKB/AKT） specific agonist］ group and SC79+IL-1β group. Western blot was used to determine the protein levels of eNOS, p-eNOS-Ser1177, AKT and p-AKT-Ser473 in the HUVECs. Chemical colorimetry was used to detect the nitric oxide （NO） content in the culture medium of HUVECs. RESULTS No statistically significant difference of p-eNOS-Ser1177 level in HUVECs treated with TNF-α and IL-6 was observed as compared with normal control group （P>0.05）, while the protein level of p-eNOS-Ser1177 in the HUVECs and the content of NO in the culture medium of HUVECs decreased significantly in IL-1β group （P<0.05）, and the protein level of p-AKT-Ser473 in the HUVECs was decreased as compared with normal control group （P<0.05）. The AKT agonist SC79 blocked the down-regulation effect of IL-1β on p-eNOS-Ser1177 level in the HUVECs and NO content in the culture medium of HUVECs （P<0.05）. CONCLUSION IL-1β down-regulates the protein level of p-eNOS-Ser1177 in HUVECs and affects the activity of eNOS, which may be involved in AKT/eNOS signaling pathway.     

洱源丛枝瑚杀虫活性成分研究

洱源丛枝瑚(Ramaria eryuanensis)石油醚萃取物经过柱层析得到单体化合物B8,该化合物对小菜蛾3龄幼虫72 h的LC50为3.375 7 mg/mL。经FAB-MS、1HNMR、13CNMR及DEPT等波谱鉴定,并与文献值对照,确定其为麦角甾-7,22-二烯-3β,5α,6β-三醇。     

通过染色体整合β-1,4-葡聚糖酶基因glu14提高绿色木霉对小麦纹枯病的防治效果

 绿色木霉LTR-2是生物防治菌株。利用来自巨大芽胞杆菌Ap25的β-1,4-葡聚糖酶基因glu14构建木霉表达载体pSilent/glu14,利用限制性内切酶介导法(REMI)转化绿色木霉LTR-2。PCR扩增及Southern杂交证实目的基因已插入木霉转化子的染色体DNA上。转化子的β-1,4-葡聚糖酶水解活性,对小麦纹枯病菌的平板抑制作用及温室防治效果较原始菌株LTR-2明显提高(P<0.01),其中转化子L-10的效果最好,平板抑制率比LTR-2提高了27.0%,温室防治效果比LTR-2提高了26.7%。本试验表明,利用REMI技术,将β-1,4-葡聚糖酶基因重组到木霉染色体DNA上,是获得高效木霉工程菌株的有效手段。     

β-折叠抗菌肽的研发及应用策略

抗生素的滥用导致全球细菌耐药性问题愈发严重,严重威胁人类、畜禽健康及畜牧业发展,功能多样且不易导致细菌产生耐药性突变的抗菌肽（antimicrobial peptides,AMPs）逐渐发展为抗生素的潜在替代品。β-折叠是抗菌肽一个主要的二级结构分类,该类肽通常由一个或多个二硫键来维系结构的稳定,现已发现许多抗菌肽具有此类结构。相比于目前研究广泛的α-螺旋AMP,它们被认为拥有更强的抗酶解能力和更高的细胞选择性。本篇综述介绍了β-折叠抗菌肽的来源和抗细菌机理,并梳理了一些常见的分子设计方法和应用策略,以期为β-折叠抗菌肽的研发应用提供新思路。     

中药成分辛弗林对胶体金免疫层析法检测β-受体激动剂的影响

旨在探究陈皮、青皮、木瓜、厚朴、红景天等5种中药材提取液以及饲喂了陈皮和青皮的猪尿液胶体金免疫层析法检测结果呈阳性的原因。建立测定猪尿及中药提取物中辛弗林的LC-MS/MS检测方法,测定陈皮、青皮的提取物和饲喂陈皮、青皮后猪尿中的辛弗林浓度。用小鼠肝微粒体孵育CGIA检测呈阳性的陈皮、青皮、木瓜和厚朴4味中药提取物。饲喂陈皮和青皮后的猪尿液检出辛弗林,浓度分别为1.36和1.65 μg·mL^-1;在陈皮和青皮的提取复溶液中检出辛弗林,浓度分别为132.6和312.7 μg·mL^-1。厚朴和木瓜两种中药提取物经过肝微粒体孵育后,在2~5 h逐渐由CGIA检测阳性转为阴性,陈皮和青皮孵育后,0~6 h的结果均为CGIA检测阳性;阴性中药组CGIA检测均呈阴性,而添加辛弗林的阳性对照组0~6 h CGIA检测均呈阳性。猪较大剂量使用陈皮和青皮会引起猪尿β-受体激动剂CGIA检测出现假阳性,该假阳性现象跟中药成分辛弗林有关,体外试验中辛弗林不易被小鼠肝微粒体代谢。     

超高效液相色谱法测定饲料中斑蝥黄和β-阿朴-8’-胡萝卜素酸乙酯含量方法的研究

为有效快速测定饲料中斑蝥黄和β-阿朴-8’-胡萝卜素酸乙酯含量,饲料经甲醇、正己烷和乙酸乙酯混合液萃取后,在氮气保护下浓缩,用甲基叔丁基醚-乙腈溶液复溶,注入超高效液相色谱仪中进行测定。结果表明:斑蝥黄浓度为1.25~10.0μg/mL,β-阿朴-8’-胡萝卜素酸乙酯为2.50~20.0μg/mL线性关系良好;斑蝥黄和β-阿朴-8’-胡萝卜素酸乙酯的检测限为0.06 mg/kg,定量限为0.20 mg/kg。斑蝥黄5个添加水平的平均回收率为72.8%~82.8%,相对标准偏差(RSD)为0.41%~1.06%;β-阿朴-8’-胡萝卜素酸乙酯5个添加水平的平均回收率为72.0%~82.8%,RSD为0.46%~1.41%。说明该方法简便、快速、灵敏度高、准确度好,适用于测定饲料中斑蝥黄和β-阿朴-8’-胡萝卜素酸乙酯含量。     

利用凝胶扩散法测定刺萼龙葵种子中β-甘露聚糖酶的活性

β-甘露聚糖酶是种子萌发过程中降解胚乳细胞壁的关键酶,明确其活性的动态变化可为揭示杂草种子的休眠萌发机制提供重要依据。以外来杂草刺萼龙葵Solanum rostratum Dunal种子为材料,建立了种子中β-甘露聚糖酶活性的检测方法—凝胶扩散法。利用凝胶扩散法对不同贮存时间及贮存条件下刺萼龙葵种子中β-甘露聚糖酶的活性进行了检测,发现贮存3年以上的种子中该酶的活性为0.03 nmol/(min·mg),显著低于贮存3年以下的种子中的酶活性0.15 nmol/(min·mg),而湿润冷藏的种子中β-甘露聚糖酶的活性为0.12 nmol/(min·mg),显著高于干燥冷藏的种子的酶活性0.02 nmol/(min·mg)。实际应用结果表明,凝胶扩散法综合了传统方法的优势,检测特异性强、灵敏度高、重复性好,可同时检测大量种子样品,具有良好的实用性。     

Uterine Expression of WNT7A and β-catenin After Induction of Oestrus in Sheep

WNT7A and β-catenin localisations and roles in regulating periimplantation ovine conceptus development under natural estrous conditions have been elaborated.However,their locations and expression patterns have not been reported under induction of oestrus.The localisation,expression and function of WNT7A and β-catenin in the uterine tissues of the early pregnant and non-pregnant sheep on days 10,12,14,16 and 18 following artificial induction of oestrus were investigated by means of in situ hybridisation,real-time RT-PCR,immuno-histochemistry and western blotting methods.WNT7A and β-catenin mRNA and protein were both restricted to the apical surfaces of the uterine luminal epithelium (LE) and glandular epithelium (GE).In pregnant sheep,protein localisation of WNT7A and β-catenin was observed both in the endometrial LE and GE.Their staining presented on day 10,increased between day 12 and day 16,and decreased on day 18.WNT7A and β-catenin mRNA and protein expression increased initially and then decreased from day 10 to day 18,peaking on day 16,and β-catenin reaching a peak on day 18 in the uterine tissues of pregnant sheep (p0.05).By contrast,no significant changes in WNT7A and β-catenin mRNA and protein expression levels were observed from day 10 to day 18 of the oestrus cycle in the uterine tissues of non-pregnant sheep (p0.05).Additionally,WNT7A and β-catenin mRNA and protein expression levels in the uterine tissues of the early pregnant sheep were significantly higher than those of non-pregnant sheep (p0.05).Treatment of endometrial epithelial cells with WNT7A increased the mRNA expressions of β-catenin,c-myc and Cyclin D1.These results provided an underlying mechanism of periimplantation ovine conceptus development under induction of oestrus.