Protein expression of cyclin D2 and p16 in proliferation and differentiation of cultured cardiac myocytes

AIM:To investigate the protein expression of cyclin D₂ and p16 in proliferation and differentiation of cultured cardiac myocytes.METHODS:One-day-old Sparague-Dawley rats were used. Cardiac myocytes(CM) were collected by a trypsin-dispersal method and cultured. Cell growth line and fluorescence activated cell sorting (FACS) were used to investigate the proliferation of CM. Ultra-thin sections were made to observe the ultrastructure of CM under transmission electron microscope. The expression of cyclin D₂ and p16 in CM were measured using immunocytochemistry and image analysis.RESULTS:①Results of cell growth line and FACS analysis showed that cultured CM could proliferate in the first 3 cultured days, but the ability decreased quickly, concomitant with differentiation. CM was obseved quiescent in cell cycle three days later. The ultrastructure of CM showed the large amount of myofilaments and mitochondrion. ②The protein expression of cyclin D₂ in 3,4,5 day CM group was 0.89 times(P＜0.05),0.80 times (P＜0.05) and 0.56 times (P＜0.01) of that in 1 day group, respectively. The expression of p16 in CM was increased during the culture process, 2,3,4,5 day group were 1.63 times, 1.72 times, 1.99 times and 2.84 times (P＜0.01) of that in 1 day group, respectively.CONCLUSION:Cultured neonatal rat cardiac myocytes could proliferate during the first 3 days after incubation, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 play the key roles in CM postnatal development. Downregulation of cyclin D₂ and upregulation of p16 may induce CM differentiation.     

施氮量对棉田主要害虫种群动态的影响

通过不同施氮肥量对棉田主要害虫种群动态影响的研究结果表明,在低、中、高3种施氮肥水平处理条件下,随着氮肥量的增加,棉田棉铃虫落卵量增加,棉苗蚜、棉叶螨种群数量减少,发生为害减轻,但棉伏蚜种群数量增加。中等施氮肥条件下,第二、三代棉铃虫幼虫数量高于低和高施氮肥处理,对棉铃虫幼虫的生长发育和存活较为有利。     

有机硒和维生素E对寿隐杂交鸡抗热应激性能的影响

通过向日粮中添加不同水平有机硒(0,0.15mg/kg)和维生素E(10,30,100IU/kg),测定热应激不同阶段试验鸡肝脏、肾脏及血清中丙二醛(MDA)含量、谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)活性。结果表明,日粮中添加有机硒和维生素E可明显降低肝脏、肾脏及血清中MDA的含量(P<0.05),提高GSH-Px活性,增强机体的抗氧化能力,提高试验鸡的抗热应激性能。     

补铬对热应激蛋鸡肾上腺组织结构的影响

试验选择处于热应激条件下的192只35周龄新罗曼褐壳蛋鸡,随机分为6组,分别饲喂添加0.10,20,30,40,50 g/t含铬量为7.410mg/kg的葡萄糖耐量因子(GTF)的玉米—豆粕型基础日粮52d。试验结束时随机抽取试验鸡36只(每组6只),取肾上腺称重并制作切片,研究有机铬对热应激蛋鸡肾上腺组织结构的影响。结果试验组肾上腺重量较对照组略有增加;热应激时蛋鸡肾上腺的肾间组织比例增大,肾间组织和嗜铬组织的细胞均有结构受损的现象;补铬使肾间组织比例减小,嗜铬细胞增多,细胞结构完好。     

谷氨酰胺对肉仔鸡脾组织结构发育的影响

160只 1日龄艾维茵肉仔鸡随机分成 4组 ,分别饲喂添加 0 % ,0 2 % ,0 4%和 0 8%谷氨酰胺的饲粮 2 8d。每周末每组取 6只鸡 ,颈静脉放血致死 ,取脾脏 ,Bouin液固定 ,制作石蜡切片 ,HE染色 ,光镜观察 ,显微摄影 ,研究基础日粮中添加谷氨酰胺对肉仔鸡脾组织结构的影响。结果显示 :添加谷氨酰胺的各试验组脾小结增多、增大 ;动脉周围淋巴鞘增厚 ;椭球增多、增大 ,细胞排列疏松 ,鞘毛细血管内皮细胞增高、增多 ;红髓比例降低 ,脾索内浆细胞和巨噬细胞增多。添加 0 8%谷氨酰胺的脾小结变化明显 ,添加 0 4%谷氨酰胺的动脉周围淋巴鞘和椭球变化明显。说明日粮中适量添加谷氨酰胺可促进脾内淋巴细胞的增殖与分化 ,从组织学角度证明 ,谷氨酰胺可促进脾的免疫应答能力 ,提高机体的免疫功能     

Nitric oxide-mediated the cardioprotection of tumor necrosis factor-alpha on cultured neonatal rat cardiomyocytes during hypoxia/reoxygenation

AIM: To investigate the role of nitric oxide synthase (NOS), soluble guanylyl cyclase (sGC) and protein kinase C (PKC) signaling in tumor necrosis factor-α (TNF-α)-induced cardioprotection against hypoxia/reoxygenation (H/R) injury. METHODS: Neonatal rat ventricular myocytes were pretreated with TNF-α or sodium nitroprusside (SNP) or L-arginine (L-Arg), respectively, for 12 h and then subjected to continuous hypoxia for 12 h, followed by reoxygenation for 6 h. The manganese superoxide dismutase (Mn-SOD) activity of the cells was measured after H/R. Myocyte injury was determined by the release of lactic dehydrogenase (LDH). RESULTS: TNF-α (10⁵ U/L) significantly increased the Mn-SOD activity and decreased release of LDH from ventricular myocytes. The cardioprotection against H/R injury was induced by the pretreatment with SNP (5 μmol/L) or L-Arg (5 mmol/L), which was blocked by ODQ (10 μmol/L), the specific sGC inhibitor, and Chel (5 μmol/L), the specific PKC inhibitor. Pretreatment with L-NAME (100 μmol/L), ODQ, Chel, antoxidant 2-MPG (400 μmol/L) or tyrosine kinase inhibitor genistein (50 μmol/L) attenuated the increased Mn-SOD activity and reduced LDH level induced by TNF-α. CONCLUSION: The results suggest that NO may play a role in TNF-α-induced cardioprotection, which is mediated by sGC and PKC.     

金龟子绿僵菌固态培养生物变量优化研究

金龟子绿僵菌属于真菌类生物杀虫剂,本文研究了接种量、种龄、菌种代数等生物因素对金龟子绿缰菌生长及产孢量的影响,并对液—固两步发酵工艺进行了研究。经过优化,得到金龟子绿僵菌产孢的最佳生物参数是:接种量为2.5g/100g、种龄为5-6天;采用固体二代种,培养6天,孢子产量可达1.531×10~(10)孢子/g干培养基。     

果寡糖对猪粪便细菌群作用下L-色氨酸代谢的影响

本试验研究了果寡糖对猪粪便细菌群作用下L -色氨酸代谢的影响。将 10 % (W/V)粪水厌氧分装于无菌瓶中 ,分成 4组 :1组 (对照 )添加 2 5 0 μmol/LL -色氨酸 ;2～ 4组 :分别在对照组的基础上添加 0 .5 % ,1.0 %和 1.5 %果寡糖。 38℃、厌氧培养。在培养期的不同时间点 (0 ,2 ,4 ,6 ,8,12 ,18,2 4h)分别从各瓶中取 1mL培养液 ,分析各培养液中吲哚类物质含量。培养 2 4h后 ,分析各培养液中细菌学指标和pH。结果表明 :含L -色氨酸的体外培养液中添加 0 .5 % ,1.0 %和 1.5 %果寡糖 ,培养 2 4h后 ,使粪臭素浓度、吲哚 - 3-乙酸峰值和pH值均显著降低。添加1.0 %和 1.5 %果寡糖显著降低色氨酸降解率和粪臭素相对产率 ,显著提高吲哚相对产率 ,显著降低梭菌和大肠杆菌数 ,显著增加双歧杆菌和总厌氧菌数。     

马杜霉素发酵法生产工艺的研究

马杜霉素是一种应用广泛的兽用抗球虫抗生素.以马杜霉拉放线菌(Actinomadura Yumanense sp.)ZU-M3为生产菌株,通过摇瓶实验获得较适宜的发酵条件:发酵培养基含葡萄糖7%,豆饼粉1.2%,玉米浆1.2%,NaCl 0.3%,CaCO3 0.1%,K2HPO3 0.03%,Fe2(SO4)3 0.01%,初始pH为6.6,发酵温度为34 ℃.用3L发酵罐做放大试验,通气速率为0.12 m3/h,搅拌速率600 r/min,发酵8 d 菌液发酵单位达到8 336 μg/mL.该研究结果对于大规模的工业化生产具有指导意义.     

大熊猫犬冠状病毒的分离与鉴定

以犬肾传代细胞(MDCK)从1只病死大熊猫肝脏中分离到1株病毒,命名为DXMV。电镜观察磷钨酸负染的病毒细胞培养物，发现冠状病毒样粒子，超薄切片上可见细胞浆内病毒包涵体。理化学和生物学鉴定结果表明，DXMV核酸类型为RNA，对氯仿、乙醚敏感，可耐受1％胰蛋白酶的作用，具有一定的耐酸性和耐热性，可在MDCK、FCWF、F81细胞中增殖，无血凝性和血吸附特性。血清中和试验结果表明，DXMV可被犬冠状病毒阳性血清中和。采用犬冠状病毒间接荧光抗体法染色DXMV细胞飞片，可在细胞浆内见到特异性荧光。以冠状病毒通用引物和犬冠状病毒特异性引物，可从大熊猫肝脏和不同代次的病毒细胞培养物中扩增出与预期值251bp和515bp大小相同的核苷酸片段。由此说明，该病毒为犬冠状病毒，大熊猫可被犬冠状病毒感染。