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精原干细胞(spermatogonial stem cells,SSCs)富集纯化是利用SSCs进行基因修饰新方法等研究的前提基础。采用免疫磁珠分选法,使用干细胞抗体CD90.2进行小鼠SSCs的纯化富集,并采用流式细胞分析法和定量PCR验证了磁珠分选效率。流式细胞分析结果:免疫磁珠分选后SSCs纯度为50.11%。荧光定量PCR检测结果:磁珠分选后支持细胞特异表达基因 GATA4 显著下调(6倍)、SSCs表达基因 GFRα-1 上调(6.5倍)、生殖干细胞特异表达基因 OCT4 极显著上调(5.9倍),3个基因相对表达量的变化说明,免疫磁珠分选效率为6倍。流式细胞分析法所产生的偏差可能是受到了未解离磁珠及SSCs本身转基因荧光的影响。 相似文献
103.
Background
In the past decades the rapid growth of molecular diagnostics (based on either traditional PCR or isothermal amplification technologies) meet the demand for fast and accurate testing. Although isothermal amplification technologies have the advantages of low cost requirements for instruments, the further improvement on sensitivity, speed and robustness is a prerequisite for the applications in rapid pathogen detection, especially at point-of-care diagnostics. Here, we describe and explore several strategies to improve one of the isothermal technologies, helicase-dependent amplification (HDA). 相似文献104.
Su K Tian Y Wang J Shi W Luo D Liu J Tong Z Wu J Zhang J Wei L 《DNA and cell biology》2012,31(6):1078-1087
Metastasis is the leading cause of death in breast cancer patients. Recent evidence suggests that inflammation-related cytokine tumor necrosis factor-alpha (TNF-α) is implicated in tumor invasion and metastasis, but the mechanism of its involvement remains elusive. In this study, we employed MCF-7 breast cancer cells as an experimental model to demonstrate that TNF-α inhibits breast cancer cell adhesion and cell proliferation through hypoxia inducible factor-1alpha (HIF-1α) mediated suppression of vasodilator-stimulated phosphoprotein (VASP). We observed that TNF-α treatment attenuated the adhesion and proliferation of MCF-7 cells it also dramatically increased HIF-1α expression and decreased VASP expression. Through a variety of approaches, including promoter assay, electrophoretic mobility shift assay (EMSA), and chromatin immunoprecipitation (ChIP), we identified VASP as a direct target gene of HIF-1α. In addition, we confirmed that HIF-1α mediated the repression of VASP expression by TNF-α in MCF-7 cells. We also demonstrated that exogenous VASP expression or knockdown of HIF-1α relieved TNF-α induced inhibition of cell adhesion and proliferation. We identified a novel TNF-α/HIF-1α/VASP axis in which HIF-1α acts downstream of TNF-α to inhibit VASP expression and modulate the adhesion and proliferation of breast cancer cells. These data provide new insight into the potential anti-tumor effects of TNF-α. 相似文献
105.
Li-Li Zhao Tong Zhang Li-Wei Zhuang Bing-Zhu Yan Rui-Feng Wang Bing-Rong Liu 《Molecular biology reports》2014,41(7):4697-4704
Pancreatic cancer is a uniformly lethal disease that can be difficult to diagnose at its early stage. Thus, our present study aimed to explore the underlying mechanism and identify new targets for this disease. The data GSE16515, including 36 tumor and 16 normal samples were available from Gene Expression Omnibus. Differentially expressed genes (DEGs) were screened out using Robust Multichip Averaging and LIMMA package. Moreover, gene ontology and pathway enrichment analyses were performed to DEGs. Followed with protein–protein interaction (PPI) network construction by STRING and Cytoscape, module analysis was conducted using ClusterONE. Finally, based on PubMed, text mining about these DEGs was carried out. Total 274 up-regulated and 93 down-regulated genes were identified as the common DEGs and these genes were discovered significantly enriched in cell adhesion and extracellular region terms, as well as ECM-receptor interaction pathway. In addition, five modules were screened out from the up-regulated PPI network with none in down-regulated network. Finally, the up-regulated genes, including MIA, MET and CEACAMS, and down-regulated genes, such as FGF, INS and LAPP, had the most references in text mining analysis. Our findings demonstrate that the up- and down-regulated genes play important roles in pancreatic cancer development and might be new targets for the therapy. 相似文献
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内源植物激素与光敏核不育水稻农垦58S育性的关系 总被引:21,自引:1,他引:21
采用气相色谱-质谱联用选择离子检测(GC-MS-SIM)技术,定性、定量分析了光敏核不育水稻农垦58S(Oryza sativa L. subsp. japonica)及对照品种“农垦58”在长(LD)、短(SD)日照处理下,不同发育时期顶端全展叶片中内源IAA、ABA 和生殖器官中内源IAA、ABA、GA1、GA4 的含量变化。实验结果表明:在LD 处理下,农垦58S雌雄蕊形成期的叶片和花粉母细胞形成期的幼穗中IAA 相继发生亏缺;农垦58S-LD花粉母细胞形成期、单核期和扬花期,生殖器官中内源ABA 均低于农垦58S-SD 的含量,“农垦58”则与此相反;扬花期不育花药中内源GA1 和GA4 含量剧减。据此认为:生殖器官中早期IAA 的亏损、ABA 含量剧减伴随着抗逆性能的减弱及GAS的不足共同导致了农垦58S的雄性败育 相似文献
110.
应用数量分类(TWINSPAN)和排序(DCA)方法,对新疆博格达山苔藓植被进行了多元分析。TWINSPAN结果表明,该地区苔藓植被可分为6个主要群落类型,即牛角藓-真藓(Cratoneuron-Bryum)群落、扭口藓-墙藓(Barbula-Tortula)群落、提灯藓-细罗藓(Mnium-Leskeella)群落、对叶藓-提灯藓(Distichium_Mnium)群落、墙藓-连轴藓(Tortula_Schistidium)群落、扭口藓-紫萼藓(Barbula_Grimmia)群落,它们分别分布于山地荒 相似文献