玻璃化冷冻和程序化冷冻方法对人类卵母细胞骨架及其发育潜能的影响

目的探讨玻璃化冷冻和程序化冷冻对人卵母细胞纺锤体定位、细胞骨架及其发育潜能的影响。方法将第2日发育为M_II卵母细胞随机分为对照组、程序化冻融组、玻璃化冻融组(解冻0 h、1 h、3 h)。应用液晶偏振光显微镜(Polscope)成像系统观察卵母细胞纺锤体与第一极体(Pb)的夹角、表面积、卵透明带内层光阻值和外层光阻值。采用扫描电子显微镜和透射电子显微镜观察卵母细胞的表面和内部超微结构。统计2种冻融方法对卵母细胞发育潜能的影响。结果对照组、程序化冷冻解冻培养3 h组和玻璃化冷冻解冻后培养0 h、1 h、3 h组中的纺锤体可见率分别为92.4%、56.4%、11.2%、24.8%、61.1%。与程序化冻融组相比,玻璃化冷冻解冻培养3 h后卵母细胞中纺锤体与Pb的夹角更小(37.3°与68°,P=0.023)。对照组、程序化冻融组和玻璃化冻融后培养3 h组中卵母细胞的纺锤体面积、卵透明带内层光阻值和透明带外层光阻值差异无统计学意义(P>0.05)。与程序化冻融组相比,玻璃化冻融后培养3 h组中卵母细胞表面突起丰富,微绒毛形态较为正常,倒伏在细胞表面,卵透明带边界较为清晰,与对照组较为接近。程序化冻融组的正常受精率(65.7%)明显低于对照组(79.2%,P=0.041),而卵裂率和囊胚形成率与对照组和玻璃化冻融组差异无统计学意义(P>0.05)。玻璃化冻融后培养3 h组中正常受精率、卵裂率、囊胚形成率与对照组相比,差异无统计学意义(P>0.05)。结论相比程序化冷冻,玻璃化冷冻对卵母细胞纺锤体和卵透明带的损伤及对卵母细胞的发育潜能的影响都较小,可以作为卵母细胞冷冻的一种有效方法。国家自然科学青年基金(81701419);;国家自然科学基金面上项目(81571418)~

Optimal Fermentation Conditions for Melanin Production by Streptomyces G-HD-4

为提高链霉菌g-Hd-4的黑色素产生量,以l-酪氨酸为底物,对该菌种产黑色素的工艺条件进行研究,通过单因素试验和均匀设计试验,得到最佳产黑色素的发酵条件为:马铃薯15g/100Ml、乳糖2.5g/100Ml、干酪素2.07g/100Ml、MgSO40.11g/100Ml、kH2PO40.25g/100Ml、l-酪氨酸0.25g/100Ml、以接种量为6%(体积分数),培养基起始PH值为6.0,装液量为25Ml,28℃,150r/MIn振荡培养5d,黑色素产量可达(13.4±0.05)g/l,比在基础培养基中的黑色素产量(5.78g/l)提高约2.3倍。In order to improve melanin production by Streptomyces G-HD-4 fermentation, the optimal conditions for fermenting L-tyrosine by this strain were investigate by single factor and uniform design methods.The optimal medium compositions (on the basis of 100 mL medium) were 15 g potato, 2.5 g lactose, 2.07 g casein, 0.25 g KH2PO4, 0.11 g MgSO4, and 0.25 g L-tyrosine.The optimal fermentation process was that 25 mL of medium was contained in a 250 mL flask for 5-day fermentation at initial pH 6.0, 28 ℃ and 150 r/min shaking speed.As a result, the maximum melanin yield reached up to (13.4 ± 0.05) g/L, approximately 2.3 times higher than that obtained using preoptimization fermentation medium

Autophagy Inhibition by Chloroquine Sensitizes Cervical Cancer SiHa Cells to CPT Treatment

自噬诱导是肿瘤细胞对化疗药物抵抗性的原因之一,该研究探讨溶酶体抑制剂氯喹对喜树碱(camptothecin,CPT)诱导的宫颈癌细胞Si Ha死亡的增敏效果。CPT和/或氯喹处理宫颈癌Si Ha细胞,MTT法检测细胞增殖,DAPI和TUNEL染色观察细胞凋亡,Western blot和免疫荧光检测自噬及凋亡相关蛋白。结果发现,CPT处理后,Si Ha细胞MAP1LC3B荧光点和LC3II(microtubuleassociated protein light chain 3II)蛋白水平增加,p62荧光点和蛋白质水平则减少;而采用氯喹特异抑制自噬后,可明显提高CPT诱导的细胞凋亡、caspase-9的激活和PARP(poly ADP-ribose polymerase)的切割,而全长caspase-2水平显著下降。以上结果提示,氯喹可通过抑制细胞自噬而增强宫颈癌细胞株Si Ha对CPT诱导细胞凋亡的敏感性。The autophagy induction is one of the reasons for the resistance of tumor cells to chemotherapy drugs. In this study, the enhanced sensitivity of cervical cancer Si Ha cells to camptothecin(CPT)-induced cell death by chloroquine(a lysosome inhibitor) was investigated. The cell viability was detected by MTT assay, meanwhile, apoptosis was observed by DAPI and TUNEL, autophagy related proteins and apoptosis proteins were analyzed by immunofluorescence(IF) staining and Western blot in Si Ha cells after CPT treatment alone or combined with chloroquine. The results found that in Si Ha cells with CPT treatment the autophagy related protein LC3 foci and microtubule-associated protein light chain 3II(LC3II) protein level was increased, but p62 foci and protein level was decreased. When autophagy was inhibited by chloroquine, the CPT-induced apoptosis was obviously enhanced, and caspase-9 was activated and PARP was cleaved, but full length caspase-2 was decreased. Taken together, these results indicated that the inhibition of autophagy by chloroquine could sensitize cervical cancer Si Ha cells to CPT inducing cell apoptosis.山西省自然科学基金(批准号:2014021037-9);; 山西医科大学汾阳学院博士启动基金(批准号:1301);山西医科大学汾阳学院科研项目基金(批准号:1422)资助的课题~

Size Distribution of Particle and Polycyclic Aromatic Hydrocarbons in Particle Emissions from Simulated Emission Sources

采用再悬浮箱模拟得到不同粒径的烹调油烟、生物质和塑料燃烧烟尘、汽车尾气和发电机烟气等颗粒,并用gC/MS对不同粒径颗粒中18种多环芳烃(PAHS)进行了分析.结果表明,烹调油烟中颗粒物存在0.44~1.0μM和2.5~10μM两个峰值,稻草和木材燃烧排放烟尘只有0.44~1.0μM一个峰值,塑料燃烧排放烟尘的峰值不明显,汽车尾气尘因含有大量的水汽导致其粒径峰值出现在2.5~10μM,而发电机排放的烟尘约93%集中在≤2.5μM的粒径范围.烹调油烟和汽车尾气尘中低环数PAHS在2.5~10μM范围内的峰值明显;随环数增加,0.44~1.0μM范围内的峰值变得明显;不同排放源亚微米颗粒中单一PAH占全部颗粒态中该PAH的比例都呈现随分子量的增大而增大的趋势.烹调油烟和燃烧排放颗粒中PAHS的组成以菲占主导,但汽车尾气和发电机烟尘中含量最高的PAHS分别是萘和苯并[g,H,I]苝.来源特征比值的比较显示,烹调油烟与生物质燃烧颗粒中PAHS的源特征较为接近,但两者都不同于汽车尾气和发电机烟尘.Particles from cooking lampblack,biomass and plastics burning smoke,gasoline vehicular exhausts and gasoline generator exhausts were prepared in a resuspension test chamber and collected using a cascade MOUDI impactor.A total of 18 polycyclic aromatic hydrocarbons( PAHs) associated with particles were analyzed by GC-MS.The results showed that there were two peaks in the range of 0.44-1.0 μm and 2.5-10 μm for cooking lampblack,and only one peak in the range of 0.44-1.0 μm for straw and wood burning smoke.But there were no clear peak for plastics burning smoke.The peak for gasoline vehicular exhausts was found in the range of 2.5-10 μm due to the influence of water vapor associated with particles,while the particles from gasoline generator exhausts were mainly in the range of ≤2.5 μm( accounting for 93% of the total mass).The peak in 2.5-10 μm was clear for cooking lampblack and gasoline vehicular exhausts.The peak in the range of 0.44-1.0 μm became more and more apparent with the increase of PAHs molecular weight.The fraction of PAH on particles less than 1.0 μm to that on the total particles increased along with PAH's molecular weight.Phenanthrene was the dominant compound for cooking lampblack and combustion smoke,while gasoline vehicular exhausts and generator exhausts were characterized with significantly high levels of naphthalene and benzo[g,h,i]perylene, respectively.The distribution of source characteristic ratios indicated that PAHs from cooking lampblack and biomass burning were close and they were different from those of vehicular exhausts and generator exhausts.国家自然科学基金项目(40971257;41171365); 环境保护公益性行业科研专项(201009004

The preparation of 238U targets on 2mm Al foils

Preparation of 238U on 2μm Al foils by molecular plating technique is described. In order to obtain optimum conditions for deposition, several parameters influencing the quality of layers such as current density, distance between the anode and cathode and the deposition time were investigated. The target thickness was determined by spectrophotometry. The uniformity and morphology of the target surface were studied by means of scanning electron microscopy, energy dispersive X-ray spectrometry and Infrared spectra. The results show that uranium is deposited in its oxide or hydroxide form uniformly and adherently onto the foil

Primary Research on the Roles of VMP1 in TNF-Induced Cell Death

为了深入了解小鼠泡膜蛋白VMP1的生物学功能,我们采用免疫荧光染色、siRNA干扰技术、流式细胞术等方法对VMP1在TNF死亡信号通路中的作用进行研究.结果显示TNF的刺激可导致VMP1从L929细胞的细胞浆转移到细胞核,提示VMP1可能在TNF信号通路中起重要作用;用特异性的siRNA抑制L929细胞中VMP1的表达,细胞则表现出对TNF的明显抗性,证明VMP1参与了TNF诱导的细胞死亡信号通路.作者初步探索了VMP1与TNF死亡信号通路的关系,为进一步研究VMP1的生物学功能打下良好基础.A series of biological methods,such as immunofluorescent staining,RNA interference and FCM,were employed to investigate the biological functions of VMP1.The results show that TNF treatment can lead to translocation of VMP1 from cytoplasm to nucleus in L929 cell line,indicating that VMP1 may play an important role in TNF signal transduction pathway.Knock-down of endogenous VMP1 endows L929 cell obvious resistance against TNF stimulation,further demonstrating that VMP1 is involved in TNF-induced cell death.We firstly uncover the relationship between VMP1 and TNF signaling pathway,which may be a good hint for further investigetion.国家自然科学基金面上项目(30500273);; 厦门市科技计划项目(3502Z20055004)资

CSpace机构知识库影音资源支持能力扩展研究与实践

[目的]提出机构知识库影音支持能力扩展方向，实现CSpace机构知识库影音支持能力扩展。 [应用背景]影音知识资源在机构产出中所占比例不断增长，扩展机构知识库影音支持能力，可更好地的揭示、发现影音知识资源，挖掘和利用其学术研究价值和潜力。 [方法]分析用户的应用需求和国内外机构知识库影音支持服务的发展趋势，构建机构知识库影音资源支持功能扩展框架，选择其中的关键技术和方法搭建实验平台，探索将其应用于CSpace系统的可行性。 [结果]实现了影音格式转换、视频场景分析和具有场景导航功能的播放器。 [结论]影音转码稳定性和效率较好，其他影音支持功能离实用还存在一定距离，将影音格式转换技术应用于CSpace机构知识库系统中，扩展了机构知识库的影音支持服务。<br /