Effects of melatonin on gut motility

AIM: To investigate the influence of melatonin at different concentrations on the gut motility. METHODS: Male C57BL mice were used in the study. The mice were intraperitoneally injected with melatonin at doses of 1 mg/kg, 5 mg/kg, 50 mg/kg and 75 mg/kg. The colon bead expulsive time was measured at 15 min and 45 min after injection. The melatonin antagonist luzindole at a dose of 5 mg/kg was used before injection of 75mg/kg melatonin. The gut transit time was also recorded with oral Evens blue after injection of melatonin. RESULTS: Compared with control group, melatonin at the dose of 1 mg/kg at 15 min decreased the colon bead expulsive time. However, melatonin at high dose (75 mg/kg) prolonged the colon bead expulsive time compared with the control, which was effectively blocked by melatonin antagonist luzindole. Compared with control group, injection of melatonin at the dose of 5 mg/kg at 45 min decreased the colon bead expulsive time. However, melatonin at high dose (75 mg/kg) prolonged the colon bead expulsive time. No difference of colon bead expulsive time between using 1 mg/kg and 50 mg/kg melatonin was observed. In whole gut transit time test, after injection of melatonin at the doses of 1 mg/kg and 5 mg/kg, the time was decreased. The expulsive time of fecal pellet with Evens blue was shorten as compared with control group. However, the gut transit time was significantly prolonged after high dose of melatonin (75 mg/kg) was used. CONCLUSION: Low dose of melatonin increases while high dose decreases the gut motility. Melatonin antagonist luzindole effectively blocks the latter effect.     

Effect of ROCK1 and ROCK2 silencing by shRNA on hypoxia-induced apoptosis of rat cardiomyocytes

AIM: To investigate the effects of Rho-associated coiled-coil protein kinase-1 (ROCK1) and ROCK2 on apoptosis induced by hypoxia in rat cardiomyocytes. METHODS: Rat cardiomyocytes were cultured primarily and identified using an antibody targeting α-actin of striated muscle. ROCK1-shRNA and ROCK2-shRNA were transiently transfected into the cells by liposome. After 48 h, these cells were subject to hypoxia for 6 h. The cells were divided into 5 groups: blank control group, hypoxia group, hypoxia+negative control shRNA group, hypoxia+ROCK1-shRNA group and hypoxia+ROCK2-shRNA group. The beating frequency and rhythm of the cardiomyocytes were assessed by microscopy. The activity of lactate dehydrogenase (LDH) in the cell culture supernatants was detected by automatic biochemical analyzer. The cell survival rate was analyzed by the method of MTT. The cell apoptotic rate was assessed by flow cytometry. Western blotting was used to determine the expression of ROCK1, ROCK2, caspase-3 and p-PI3K. RESULTS: The primary culture of the cardiomyocytes was successful. Western blotting results showed that the transfection of ROCK1-shRNA or ROCK2-shRNA decreased the expression of ROCK1 or ROCK2 in the cardiomyocytes. Hypoxia slowed down the beat frequency of the cardiomyocytes, also made the rhythm disorder. Hypoxia increased the release of LDH and decreased the cell survival rate. Flow cytometry results showed that hypoxia increased the cell apoptotic rate. Hypoxia increased the expression of caspase-3 and decreased the expression of p-PI3K. Transfection of ROCK1-shRNA and ROCK2-shRNA into the cardiomyocytes reduced all the effects of hypoxia mentioned above. CONCLUSION: Down-regulation of ROCK1 and ROCK2 expression suppresses the apoptosis of rat cardiomyocytes induced by hypoxia. The mechanism is associated with the inhibition of caspase-3 activation and the up-regulation of p-PI3K expression.     

山苍子的栽培和利用(上篇)

山苍子是一种经济价值很高的珍贵天然木本芳香油料树种。具有投资少、结实早、收效快的特点。科学采种、及时加工、薄膜育苗,对壮苗培育、提高优质出油率,至关重要。     

基于模糊模式识别理论的富营养化评价模型及应用

依据水体富营养化及其程度属于模糊概念这一客观实际,基于模糊模式识别理论,以叶绿素a、总磷、总氮、透明度和高锰酸盐指数作为评价指标,尝试建立富营养化评价的模糊模式识别模型。并以南方某大型水库为例,按年、汛期和非汛期3个阶段开展了富营养化评价研究。评价结果表明2005-2008年年均富营养化特征值分别为2.829 0、2.717 7、2.844 9、2.535 4,汛期分别为2.913 8、2.731 4、2.971 7、2.444 7,非汛期分别为2.748 3、2.712 5、2.713 8、2.648 8。可见该水库正处于由中营养向富营养过渡阶段,实例同时也表明该方法用于水体富营养化评价是合理、可行的。     

副猪嗜血杆菌S-核糖基高半胱氨酸酶基因序列分析与推导蛋白三维结构的分子模拟

利用PCR方法获得副猪嗜血杆菌S-核糖基高半胱氨酸酶(luxS)基因全长DNA,将PCR纯化产物与pMD18-T载体连接并转化E.coil DH5α菌株,重组阳性质粒测序并采用生物信息学软件对所推导的氨基酸序列进行三维结构分析。结果表明,该基因全长510bp,并与GenBank中登录的其他5株菌株luxS基因完整参考序列进行比较,同源性均在70%以上。用ExPAsy软件包预测了推导蛋白的特性,运用Swiss-PDB viewer软件的SWISS-Model处理器,并利用同源建模的思想建立HPS-luxS的三维结构。拉马钱德兰图证明,构建的luxS蛋白的空间结构是合理的。     

牛叠朊编码基因缺失突变体的构建及在大肠杆菌中的表达

本实验室已制备出多株针对牛叠朊的单克隆抗体,为充分鉴定这些单克隆抗体针对的抗原表位,通过基因克隆表达的策略获取牛叠朊基因缺失突变体的重组蛋白,以期为单克隆抗体的表位鉴定提供物质材料。经过对牛成熟叠朊编码基因及预测蛋白结构特征的分析,设计表达9个缺失突变体的引物。以PCR扩增出叠朊基因的缺失突变体,与pET-30a（＋）表达载体连接后转入E.coli DH5α中,经双酶切和测序鉴定后将重组质粒转入E.coliJM109和E.coli BL21表达宿主菌中,经IPTG诱导表达后,以SDS-PAGE、Western blot和ELISA检测表达产物的相对分子质量、相对表达量和反应原性。结果表明,成功构建了9个牛叠朊编码基因的缺失突变体,通过IPTG的诱导表达,其中有6个缺失突变体被大肠杆菌高效表达,并且具有较好的反应原性,这为其单克隆抗体表位的进一步鉴定奠定了物质基础。     

三聚氰酸和三聚氰胺对肉鸡生长性能的影响及其组织残留的研究

本试验旨在探讨饲粮中添加不同水平的三聚氰酸(cyanuric acid,CYA)和/或三聚氰胺(melamine,MEL)对肉鸡生长性能的影响及其在血浆、组织中的残留.试验采用2×5两因子设计,1 200只1日龄科宝500肉鸡公雏随机分为10个处理,每个处理6个重复,每个重复20只鸡.CYA设5个水平:0、10、20、...     

N-乙酰半胱氨酸对脂多糖刺激仔猪肌肉能量状况的影响

本试验研究了N-乙酰半胱氨酸(NAC)对脂多糖(LPS)刺激仔猪肌肉能量状况的影响.选取18头杜洛克×长白×大白健康仔猪,体重(11.58±0.26)KG,随机分成3个处理(对照组、LPS组和NAC组),每个处理6个重复,每个重复1头猪.对照组和LPS组饲喂基础饲粮,NAC组饲喂基础饲粮+500 mg/kg NAC,L...     

T-2毒素研究进展

T-2毒素是污染我国饲料的主要霉菌毒素,主要危害动物的造血组织和免疫器官,引起出血性综合征,出现白细胞减少、贫血,使胃肠道功能受损等,对畜禽健康和畜牧业生产的危害很大.本文主要从T-2毒素的一般性质、产毒菌株、毒性效应、毒性机制及预防控制等方面作了论述.     

同时检测猪圆环病毒2型、猪细小病毒和伪狂犬病毒连接酶检测反应-PCR基因芯片检测方法的建立

为快速、灵敏、准确地同时检测和鉴别猪圆环病毒2型(PCV2)、猪细小病毒(PPV)和伪狂犬病毒(PRV)的方法,本研究采用连接酶检测反应(LDR)-PCR和基因芯片技术建立一种新型检测方法.首先在3种病毒的保守区内分别设计一对LDR探针,两端各连接一段通用序列,依次进行LDR、通用引物荧光标记扩增和芯片杂交,同时比较引物标记和Cy5 -dCTP标记方法的灵敏度.结果表明该方法可以特异地检测PCV2、PPV和PRV3种病毒,而对牛病毒性腹泻病毒、猪传染性胃肠炎病毒、猪流行性腹泻病毒、猪繁殖与呼吸障碍综合征病毒、猪瘟病毒、乙型脑炎病毒、猪圆环病毒1型检测结果均为阴性;对3种病毒的最低检测限少于10个拷贝;Cy5-dCTP标记检测的灵敏度显著高于引物标记.利用建立的方法对41例临床样品进行检测,与普通PCR检测结果符合率为97.6％～100％.该方法的建立为基础研究和临床应用提供了技术平台.