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81.
Yan Ding Hua Yang Jing Bo Feng Ying Qiu Dong Sheng Li Yi Zeng 《In vitro cellular & developmental biology. Animal》2013,49(10):771-777
Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) hold great potential for their therapeutic use in various clinical diseases. Many publications have reported on human blood-derived alternatives to animal serum for culturing mesenchymal stem cells, such as human serum, allogenic umbilical cord blood serum, and human platelet derivatives. However, it is not clear whether human umbilical cord blood plasma (UCBP), as the surplusage of umbilical cord blood mesenchymal stem cell extraction, could be used. In this study, in order to make the best of umbilical cord blood, the human UCBP was dialyzed to replace fetal bovine serum (FBS) in the culture medium. hUC-MSCs were cultured in the new medium. Cell growth rate, specific biomarkers, and differentiation properties were detected to characterize the cell proliferation and MSC-specific properties. The hUC-MSCs cultured in such derived medium were verified with proliferation rate, cluster differentiation markers, cell cycle, as well as differentiation capabilities. Such dialyzed human UCBP is fully comparable with, if not superior to, FBS in deriving and culturing hUC-MSCs. 相似文献
82.
Min Wu Qi Shen Yong Yang Sheng Zhang Wen Qu Jing Chen Hongying Sun Shuqing Chen 《Journal of industrial microbiology & biotechnology》2013,40(6):589-599
Human serum albumin (HSA) and human parathyroid hormone (1-34) [PTH (1-34)] fusion protein [HSA/PTH (1-34)] is a promising long-acting form of PTH (1-34) for osteoporosis treatment. Secretory expression of intact HSA/PTH (1-34) in Pichia pastoris GS115 was accompanied by two degradation fragments, with molecular weights around 66 kDa, in addition to the well-known ~45 kDa HSA-truncated fragment, resulting in a low yield of intact protein. In this study, two internal cleavage sites were identified in the PTH (1-34) portion of the fusion protein by Western Blot analysis. To minimize proteolytic cleavages, several protease genes including PEP4 (encoding proteinase A), PRB1 (proteinase B) and seven YPSs genes (yapsin family members) were knocked out respectively by disruption of the individual genes and the selective combinations. Reduced degradation was observed by single disruption of either PEP4 gene or YPS1 gene, and the lowest level of degradation was observed in a pep4△yps1△ double disruptant. After 72 h of induction, more than 80 % of the HSA/PTH (1-34) secreted by the pep4△yps1△ double disruptant remained intact, in comparison to only 30 % with the wild-type strain. 相似文献
83.
CD3ε和PMA对fas基因转录调控作用的研究 总被引:2,自引:0,他引:2
研究了CD3ε和PMA对细胞凋亡基因fas的转录调控作用.根据已知的人fas基因5'上游序列设计引物,用PCR法从人胸腺细胞基因组DNA中扩增出fas5'上游长为446bp的启动子片段.将此片段定向克隆到以虫荧光素酶为报告基因的真核细胞表达载体pXP2中.经限制性内切酶酶切鉴定及序列测定分析表明此重组表达质粒DNA的结构和序列正确.用此质粒(pXP2-fasup)瞬时转染人JurkatT淋巴细胞,分析报告基因的表达水平.结果表明fas基因上游-506到-60的区域有弱启动子活性,约是阴性对照的1.5倍.抗CD3ε抗体和PMA处理均可增强fas启动子的活性,促进报告基因的表达,其虫荧光素酶活性分别是未经处理的pXP2-fasup转染细胞的1.7倍和3.3倍,但二者没有协同作用.PKC的抑制剂staurosporine和PTK的抑制剂herbimycinA可抑制PMA诱导的fas基因转录,使虫荧光素酶基因的表达降至未用PMA处理的水平.但PTK的另一种抑制剂genistein可与PMA发挥协同作用,上调fas基因的转录,使虫荧光素酶活性增加到5倍.这些结果为阐明CD3ε及PMA介导T淋巴细胞激活和凋亡的分子机制 相似文献
84.
Shen YH Song GX Liu YQ Sun W Zhou LJ Liu HL Yang R Sheng YH Qian LM Kong XQ 《Journal of bioenergetics and biomembranes》2012,44(3):317-323
Fatty acid binding protein 3 (FABP3) (also known as H-FABP) is a member of the intracellular lipid-binding protein family, and is mainly expressed in cardiac muscle tissue. The in vivo function of FABP3 is proposed to be in fatty acid metabolism, trafficking, and cell signaling. Our previous study found that FABP3 is highly regulated in patients with ventricular septal defect (VSD), and may play a significant role in the development of human VSD. In the present study, we aimed to investigate the impact of FABP3 knockdown by RNA interference (RNAi) on apoptosis and mitochondrial function of embryonic carcinoma (P19) cells. The results revealed that downregulated FABP3 expression promoted apoptosis, and resulted in mitochondrial deformation, increased mitochondrial membrane potential (MMP), and decreased intracellular ATP synthesis. In addition, the knockdown of FABP3 also led to excess intracellular ROS production. However, there was no obvious influence on the amount of mitochondrial DNA. Collectively, our results indicated that FABP3 knockdown promoted apoptosis and caused mitochondrial dysfunction in P19 cells, which might be responsible for the development of human VSD. 相似文献
85.
根癌农杆菌介导Bt基因转化水稻的研究 总被引:2,自引:0,他引:2
为了培育出无筛选标记基因的转基因水稻,试验将loxp-hpt-loxp基因与成基因连锁在-起转化水稻方法,得到loxp-hpt—loxp—Bt转基因水稻植株,再与同质的带有ere基因的水稻杂交,以定向删除潮霉素抗性筛选标记。试验表明以水稻品种“皖粳97”为供试材料,将成熟胚来源的愈伤组织用根癌农杆菌EHA105/pCAMBIA1305.1感染后,筛选出抗性愈伤组织并获得再生植株。经PCR验证,得到20棵转基因水稻植株。 相似文献
86.
目的:探讨5.12汶川地震军队救援人员创伤后应激损伤状况。方法:采用创伤后压力诊断量表、焦虑自评量表、抑郁自评量表和匹兹堡睡眠质量问卷,在地震爆发后65天左右,对107名执行运输遗体任务的官兵进行了测量。结果:PTSD症状轻度的占84%,中重度的占15%。其中症状在1个月内消失的占63%。从症状的分布来看,闯入性回忆占36.4%,回避29.9%,过度警觉29.9%,焦躁不安或易怒37.4%,入睡困难、睡眠易醒或早醒43.9%,难以集中注意力45.8%,对重要的活动兴趣下降32.7%。救援人员抑郁症状得分低于全国常模水平;焦虑自评量表得分与全国常模差异不显著。睡眠质量差的占72%,以入睡时间长和日间功能降低为主。结论:担任遗体运输的救援官兵在救援早期普遍存在创伤后应激反症状,症状程度较轻,多数在1个月内消失,焦虑和抑郁症状水平的总体状况也比较低,但普遍存在睡眠质量差的问题,并且直接影响到日间功能。 相似文献
87.
Agnes Simonyi Yan He Wenwen Sheng Albert Y. Sun W. Gibson Wood Gary A. Weisman Grace Y. Sun 《Molecular neurobiology》2010,41(2-3):73-86
Alzheimer’s disease (AD) is marked by an increase in the production of extracellular beta amyloid plaques and intracellular neurofibrillary tangles associated with a decline in brain function. Increases in oxidative stress are regarded as an early sign of AD pathophysiology, although the source of reactive oxygen species (ROS) and the mechanism(s) whereby beta amyloid peptides (Aβ) impact oxidative stress have not been adequately investigated. Recent studies provide strong evidence for the involvement of NADPH oxidase and its downstream oxidative signaling pathways in the toxic effects elicited by Aβ. ROS produced by NADPH oxidase activate multiple signaling pathways leading to neuronal excitotoxicity and glial cell-mediated inflammation. This review describes recent studies demonstrating the neurotoxic effects of Aβ in conjunction with ROS produced by NADPH oxidase and the downstream pathways leading to activation of cytosolic phospholipase A2 (PLA2) and secretory PLA2. In addition, this review also describes recent studies using botanical antioxidants to protect against oxidative damage associated with AD. Investigating the metabolic and signaling pathways involving Aβ NADPH oxidase and PLA2 can help understand the mechanisms underlying the neurodegenerative effects of oxidative stress in AD. This information should provide new therapeutic approaches for prevention of this debilitating disease. 相似文献
88.
Tamoxifen inducible cre-loxp system has been employed as a very powerful tool for cardiology research in mice. It enables researchers to control their interested gene expression in tight control in terms of tissue specification and time specification. Here, we reported that in the absence of loxp transgenes,tamoxifen injection can lead to myocardiac dysfunction 3 days after drug administration but cardiac function start recover from 2 days later. 相似文献
89.
通过对长选3S和培矮64S定长的穗颈节间细胞个数统计和细胞长度的测量,得出长穗颈不育水稻穗颈节间伸长的细胞学变化规律.自然条件下长穗颈不育水稻长选3S穗颈节间比对照培矮64S长18.0cm,其纵列厚壁细胞和薄壁细胞数分别比培矮64S多1248个和580个;厚壁细胞和薄壁细胞平均长度分别比培矮64S长23.3μm和38.3μm,尤其是中部区段的厚壁细胞和薄壁细胞分别比对照长24.8μm和48.7μm;穗颈节间薄壁细胞的长度由基部区段至中部区段呈突跃式增加,而由中部至顶部区段则呈缓慢减少的趋势.隐性长穗颈温敏核不育水稻穗颈节间伸长是由细胞数目增多和细胞长度增加双重作用所致,其中后者作用更显著. 相似文献
90.