Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.     

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.     

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。     

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.     

洞庭湖湿地虉草替代大白菜对肉兔生产性能的影响

试验采用48只青年期(40～45日龄、体重约800 g)发育良好、健康无病的新西兰肉兔,随机分为4组:1组饲喂基础日粮,2～4组分别用占日粮5%,15%和25%的虉草Phalaris arundinacea代替日粮中等量的大白菜,试验期35 d.试验结果表明:各组间兔的日增重、屠宰率、料肉比等指标差异均不显著(P＞0.05),可见虉草代替大白菜不影响兔的生长,以15%的替代量的3组增重效果最佳,经济效益最好.     

Effect of L-lysine Added in Diluent on the Fresh Semen Quality of Dorper Sheep

In order to explore the effect of different L-lysine levels added in semen dilution on the fresh Dorper sheep quality stored at liquid state,3 health Dorper ram were used to collect semen which were equal-packaged and added to solution containing different levels (0,0.1,0.2,0.3,0.4 and 0.5 g) L-lysine in 120 mL diluent and at 0,6,24,30,48,54,72 and 78 h,the sperm motility,membrane integrity and acrosome integrity were detected.The results showed that the sperm motility in groups added 0,0.1 and 0.2 g L-lysine were higher than other groups,and that in 0.4 and 0.5 g L-lysine groups decreased faster than other groups;The sperm motility in 0.5 g L-lysine group reduced to 0 at 30 h,and it reduced to 0 at 48 h in the group added 0.4 g L-lysine;The effective survival time and survival index in the group added 0.1 g L-lysine was higher than other groups (P< 0.05);The membrane integrity of 0.1 g group was the highest,that had no significant difference with control group from 0 to 54 h (P >0.05),while after 72 h the two groups had significant differences (P< 0.05).The membrane integrity in 0.4 and 0.5 g groups were the worst,and that was significant different with other groups (P< 0.05);The acrosome integrity of 0.1 g group was the highest which had no significant difference with control group (P >0.05),0.4 and 0.5 g groups were the worst,the difference between them was not significant (P >0.05),while was extremely significant difference with other groups except for 0 and 6 h (P< 0.01).The results suggested that dilution added a high concentration of L-lysine could inhibit sperm quality,when the count of L-lysine was more than 0.2 g,sperm quality was significantly decreased,adding 0.1 g L-lysine could improve Dorper sheep fresh sperm quality stored at liquid state.     

毕节特产珍稀药材圆珠半夏的HPLC指纹图谱分析

为毕节大方圆珠半夏质量标准的建立提供参考,采用HPLC法对10批不同产地及不同采收时间的圆珠半夏进行了指纹图谱研究。结果表明:测定的10批半夏样品有23个共有峰,共有峰面积为91.6%~95.8%,相对峰面积的RSD为0.12%~0.40%;共有峰的相对保留时间从0.182~1.810min,相对保留时间的RSD在0.02%~0.10%。结论,该方法具有良好的稳定性和重复性,可用于圆珠半夏化学成分分析。