Effects of β-ME and RA on expression of GFAP in mesenchymal cells derived from mouse fetal liver

AIM: To investigate the effects of β-mercaptoethanol (β-ME) and all-trans rentinal acid (RA) on glial fibrillary acidic protein (GFAP) expression in mesenchymal cells derived from mouse fetal liver in vitro. METHODS: Cells suspension from 14.5-days-old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20％ FCS and mesenchymal cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by β-ME and RA. The characteristics of treated cells were assayed by immunocytochemistry staining at 5 hours and 5 days after induction. β-actin as an internal control, GFAP gene expression of mesenchyal cells was detected with semi-quantitative RT-PCR. RESULTS: After being inducted by β-ME and RA, 80％ approximately of the cells exhibited typical neural morphology and about 85％ expressed GFAP phenotype. Semi-quantitative RT-PCR showed that mRNA expression of GFAP increased in treated cells versus untreated cells (P<0.01). CONCLUSION: GFAP expression in mesenchymal cells derived from mouse fetal liver in vitro increases after being treated with β-ME and RA.     

Meiotic competence of mouse oocytes reconstructed by replacing the germinal vesicle with a male pronucleus and somatic nucleus

The present study examines the contribution of the nucleus to meiotic competence in mouse oocytes that were reconstructed using nuclear transfer. Three types of reconstructed oocytes were produced: MP‐GV, by transplanting the male pronucleus (MP) into germinal vesicle (GV) stage oocytes; 3T3‐GV, by transplanting the nucleus of a National Institute of Health (NIH) 3T3 cell into a GV stage oocyte; and 3T3‐MII, by transplanting the nucleus of an NIH 3T3 cell into a metaphase II (MII) stage oocyte. The fusion rates differed, but not significantly, in the MP‐GV, 3T3‐GV, and 3T3‐MII groups (77, 63, 56%, respectively). Then, meiotic competence was compared in MP‐GV, 3T3‐GV and non‐manipulated GV stage oocytes as a control. Nuclear envelope breakdown occurred in all the reconstructed oocytes, as well as the control ones. The percentage of first polar body extrusion differed between the MP‐GV (100%), 3T3‐GV (72%), and control (67%) groups. DNA staining with Hoechst 33342 revealed that in the MP‐GV‐group oocytes that had reached MII stage, the chromosomes were condensed and aligned in a regular array similar to the normal metaphase plate. By contrast, in 3T3‐GV group oocytes, the condensed chromosomes were irregularly scattered in the cytoplasm. These results suggest that the donor nucleus affects meiotic competence in reconstructed oocytes.     

猪胸膜肺炎放线杆菌外膜蛋白(OMP)5′末端保守区基因的克隆、表达及其免疫活性研究

以猪胸膜肺炎放线杆菌血清7型25-4株基因组DNA为模板,PCR方法扩增外膜蛋白(OMP)5′末端保守区基因片段(OMPc),酶切及核苷酸序列分析鉴定后,与原核表达载体质粒pGEX-6P-1进行连接,构建成重组表达载体pGEX-OM-Pc,转入大肠杆菌BL21中,以IPTG进行诱导,SDS-PAGE电泳分析发现,转化了重组质粒的菌株所表达的融合蛋白相对分子量为34 kD,与实际预测相符,命名为GST-OMPc.GST亲和层析柱进行纯化,ELISA方法对纯化蛋白进行检测.结果表明:纯化蛋白GST-OMPc能够与兔抗猪胸膜肺炎放线杆菌血清7型的阳性血清反应.OMPc蛋白的成功表达为其功能的研究打下基础.     

草地土壤肥力及其营养管理的必要性

介绍了草地土壤肥力的来源,土壤基本养分对草地生产的作用,草地植物与动物对养分的需求量及缺乏症状,说明了加强草地营养管理以及维持草地生态系统营养平衡的重要性,指出增强对土壤肥力和营养管理是提高草地生产能力、保证草地生态系统平衡、保障草地可持续利用的关键技术措施.     

过氧化氢提高燕麦幼苗耐碱性的活性氧代谢和渗透调节

为探明过氧化氢(H₂O₂)提高燕麦耐碱性的作用,以‘定莜6号’幼苗为材料,采用珍珠岩栽培方法,在幼苗三叶一心期根部浇灌75 mmol·L^-1 NaHCO₃或添加二甲基硫脲(DMTU,H₂O₂淬灭剂)或抗坏血酸(ASA,H₂O₂清除剂)模拟碱胁迫,通过叶面喷施0.01 mmol·L^-1 H₂O₂来观测H₂O₂对碱胁迫下幼苗生长、活性氧代谢和渗透溶质积累的影响。结果表明:喷施H₂O₂ 能够缓解NaHCO₃胁迫对燕麦幼苗生长的抑制,降低幼苗叶片O2·-、H₂O₂和丙二醛含量,提高超氧化物歧化酶、抗坏血酸过氧化物酶等抗氧化酶活性和非酶抗氧化剂抗坏血酸、谷胱甘肽含量;使过氧化氢酶、过氧化物酶活性及渗透溶质可溶性糖、游离氨基酸和脯氨酸含量显著降低,有机酸和叶绿素含量明显提高,而可溶性蛋白质含量则变化不大;添加DMTU和ASA后有效逆转了喷施H₂O₂对NaHCO₃胁迫燕麦生长受抑和生理响应的调节。采用主成分和隶属函数分析显示,喷施H₂O₂显著提高了NaHCO₃胁迫下燕麦幼苗的综合评价值,添加DMTU和ASA完全或部分逆转了H₂O₂的作用。因此,外源H₂O₂是通过调控活性氧代谢和渗透调节来缓解碱胁迫导致的氧化伤害和生长抑制,从而增强燕麦幼苗的耐碱性。     

铅胁迫对金丝草生长及生理生化的影响

采用土培方法,探究不同梯度铅胁迫(0、1000、2000和3000 mg·kg^-1)对金丝草生长形态、体内抗氧化系统和渗透调节物质的影响。结果表明:低浓度(1000 mg·kg^-1)处理会诱导金丝草叶片过氧化物酶(POD)活性、可溶性蛋白(SP)含量和根系抗超氧阴离子自由基活力(ASAFR)、可溶性糖(SS)及脯氨酸(Pro)含量增加,使得金丝草植株总抗氧化能力(T-AOC)处于较高水平,促进了金丝草株高、叶长和生物量的增加。随胁迫浓度增加,高浓度(2000~3000 mg·kg^-1)处理下,金丝草叶片和根系丙二醛(MDA)含量迅速增加,株高、叶长、叶面积和生物量下降,生长受到抑制。但金丝草通过增强叶片和根系POD、过氧化氢酶(CAT)活性来抵御过氧化作用,提高可溶性蛋白和可溶性糖含量维持细胞正常运作,增加根系生物量占比来加强根系发育,一定程度适应了高浓度铅胁迫。综上表明,金丝草主要通过叶片和根系不同抗氧化酶差异化响应、提高渗透调节物质含量,提升金丝草植株总抗氧化能力等途径来提高Pb耐性,对Pb污染矿区植物修复有较大潜力。     

连续时间系统单位冲激响应的直接确定

目的 避开用传统方法求解冲激响应时求解系统微分方程的初始条件问题,直接在时域中确定线性时不变连续时间系统的单位冲激响应.方法 将系统的转移算子H(P)按部分分式展开成一种标准形式,推导出系统的单位冲激响应.结果 求解线性时不变连续时间系统单位冲激响应,可在时域中直接确定,无须考虑初始条件.结论 这种方法与传统方法相比较,更简便而且易于掌握.     

基于模糊层次分析法的精品课程评估

目的 多方面综合评估精品课程.方法 层次分析法和模糊综合评判法相结合.结果 构建了精品课程综合评估指标体系,综合评估了某高校的高等数学课程.结论 充分利用了两种方法的优点,避免了定性评估的不足,取得了良好的评估效果.     

数学期望不等式的应用

目的 利用微积分理论证明不等式的方法很多,探讨用概率理论证明不等式的方法,方法 首先构造一个概率密度函数,再利用数学期望不等式E（η）^2≥E^2（η）证明一个积分不等式,结果 由该积分不等式推出若干数学不等式。结论 利用概率方法可以证明某些不等式,且方法简单,关键是构造恰当的概率密度函数,再利用概率中有关不等式的性质。     

淳昌清麹酱的体外抗癌效果

目的 对从淳昌郡购买的3种不同的清麹酱,Kim Yong-Soon清麹酱(K-chungkukjang),Hyang Juk-Won清麹酱(H-chungkukjang)和Moon Ok-Rae清麹酱(M-chungkukjang)进行癌细胞体外抗癌效果评价.方法 MTT试验和细胞生长抑制试验.结果 样品浓度1.0 mg/ml下分别对AGS和HT-29癌细胞进行处理,K-清麹酱具有最高的癌细胞生长抑制效果(87% and 86%).K-清麹酱能够抑制癌细胞生长速度且呈时间依赖性.结论 K-清麹酱的外抗癌效果优于其他清麹酱.