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31.
AIM: To explore the ex vivo expansion characteristics of the endothelial progenitor cells (EPCs). METHODS: CD34+ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded at the same conditions as that for total MNC, coincubation of CD34+ and CD34- from the same donation for EPCs. In addition, we tested the effect of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis. EPCs were determined and quantified by immunocytochemistry and flow cytometry. RESULTS: Coculture of CD34+ and CD34-,total MNC led to a significant increase in the expansion of CD34+ cells compared with CD34 enrichment (P<0.05). There was a trend toward decreased apoptosis in cultures when early passage was performed once the linear cord like structures appeared. There was no significant effect on apoptosis between with VEGF and without VEGF group (P>0.05). These differentiated EPCs were stained positive for CD34+, von Willebrand factor (vWF), KDR, CD31 and incorporate acetylated low-density lipoprotein (LDL). CD34+ and AC133+cells accounted for 68.2%±6.3% (n=6) and 57.2%±9.8% (n=6) of attaching (AT) cells at day 7 of culture, respectively. CONCLUSIONS: Coculture of CD34+ and CD34- or culture of MNC enhances ex vivo expansion of EPCs. Early passage decreases apoptosis rate, VEGF has no significant effect on ex vivo expansion of EPCs. 相似文献
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AIM: To study the expression of vascular endothelial growth factor (VEGF) in inflammatory mucosa of lacrimal sac. METHODS: Immunohistochemical S-P method was used to examine the expression of VEGF in the mocusa from 12 patients with chronic dacryocystitis and 8 volunteers. RESULTS: The positive rates of VEGF expression in different parts of the mocusa were: basal lamina: 44.3%±7.6%; surface epithelium: 16.9%±4.6%; connective tissue: 15.2%±4.9%, all normal mocusa of 8 cases were negative. There was a significant difference between the two groups (P<0.01), a significant difference among each part of the chronic inflammatory mocusa of lacrimal sac. CONCLUSION: VEGF may play an important role in hyperplasia of inflammatory mucosa of lacrimal sac. 相似文献
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AIM: To observe the change of insulin receptor in rabbit kidney with acute ischemic-reperfusion injury. METHODS: 15 Japanese white rabbits were allocated randomly into control group, ischemic-reperfusion group(IR group). IR group received clamping for 1 h followed by 2 h or 48 h of reperfusion. At 2 h or 48 h after reperfusion, glucose and insulin in serum were determined. Insulin receptor in renal tissue was analyzed by radioligand binging assay(BAD). RESULTS: The level of serum glucose increased after 2 h reperfusion in 2 groups, but in IR group the value increased much more higher than those in control groups(P<0.05). Plasma insulin of IR group was significantly higher than that in control after 2 h reperfusion(P<0.05). Scatchard analysis of data resulted in curvilinear profiles, indicating that there are two classes of receptors with different affinity or the presence of a single class of receptors with a negative cooperative hormone-receptor interaction. Data analyzed for a two-site model showed that the values of Bmax1(high affinity site), Bmax2(low affinity site) and Kd1, Kd2 were significantly lower than that of control (P<0.05) after 2 h perfusion. 48 h after IR there was no difference of Bmax1, Bmax2, and Kd1 between 2 groups,but Kd2 of IR group was higher than that of control (P<0.05). CONCLUSIONS: The results indicate that the effect of intrinsic insulin decreases in the progress of the renal ischemic-reperfusion. The resulting high serum glucose may aggravate renal injury in the progress of ischemic-reperfusion. 相似文献
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猪原始生殖嵴细胞(PGCs)建系因素的研究 总被引:4,自引:3,他引:4
从五指山猪(WSZP)近交系第8~13代培育群中,先后选用21头5~10月龄青年母猪,分别于授精后25~30d采集胎儿106个,进行原始生殖嵴(PGCs)细胞分离、培养等建系技术研究。以DMEM F10(1:1)为基础培养液,按添加或不添加生长因子,将培养液分为A、B、C3种,并以STO细胞作饲养层,在38℃、5.0%CO2和湿润的气相中进行培养建系。结果获得胚胎生殖嵴细胞(EG)细胞系6个细胞株,其中1个EG细胞株传至11代、2个传至5代、1个传至4代、2个传至3代冻存。并进行了AKP染色、体外分化、冷冻-解冻复苏和嵌合体制作等鉴定研究。研究发现:不同胚龄对EG细胞建系具有一定影响,不同培养液对EG细胞建系效果不同,STO细胞饲养层的质量是建株、传代、冷冻-解冻复苏的关键因素之一。EG细胞系的初步建立,为今后筛选进入种系的EG细胞系、实施体外基因操作提供了可能。 相似文献
37.
植物育种的物理学方法 总被引:10,自引:0,他引:10
介绍了草原生态恢复中选育优良牧草种子的一些物理学方法,包括电场、磁场、物理辐射、激光、离子束和交变应力等在植物的生物学效应、诱变育种和转基因技术中的应用. 相似文献
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构建表达eae和stx1/2B的融合基因,克隆eae基因的C端280个氨基酸残基(Int280)基因部分,以正确地阅读框定向插入到含有stx1/2B融合基因的质粒,构建重组质粒,将其转化于BL21(DE3),用IPTG进行诱导表达,经SDS—PAGE电泳检测,该融合蛋白获得了高效表达。薄层扫描分析表明:目的蛋白表达量占菌体总蛋白含量的50.67%。由于该融合蛋白由eae、stx1B、stx2B等三部分抗原组成,可刺激机体产生针对紧密素和StxB的抗体,在EHEC O157亚单位疫苗设计或单克隆抗体抗制备中具有重要价值。 相似文献
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