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抑制消减杂交技术克隆晕船易感大鼠脑干差异表达基因 总被引:1,自引:1,他引:1
目的 筛选晕船易感大鼠脑干组织差异表达基因。方法 利用抑制消减杂交(SSH)技术构建晕船易感大鼠脑干组织差异表达基因 cDNA文库,斑点杂交筛选含有插入片段的阳性克隆。结果 文库中442个克隆含有插入片段,斑点杂交获得的阳性克隆测序,获得39条差异基因片段和2条全长序列基因。其中高表达16条,含未知功能基因片段3条;低表达25条,含未知功能基因片段5条。结论 SSH技术是一种高效的筛选差异表达基因的方法,本实验结果为深入研究晕船易感性机制提供了重要的线索。 相似文献
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目的:研究与验证改良碘量法快速测定地震灾区饮水消毒中有效氯含量的效果。方法:取漂白粉精片、二氯异氰尿酸钠和三氯异氰尿酸消毒液各3个样品,每一样品的有效氯含量按本法各测定5次,以此进行精密度试验;另各取10份同一漂白粉精片和二氯异氰尿酸钠消毒液,分别用本法和国标碘量法测定其有效氯含量,比较两种方法的测定结果。结果:精密度平行对照实验显示,测定3种不同含氯消毒剂有效氯含量(%),其标准差(SD)和相对标准偏差(RSD)分别为:SD=±0.27~0.42,RSD=0.73%~1.12%(漂白粉精片)、SD=±0.35~0.57,RSD=0.86%~1.40%(二氯异氰尿酸钠)和SD=±0.42~0.65,RSD=0.80%~1.25%(三氯异氰尿酸);本法与国标碘量法比较,两者测定结果的均值经t检验无显著性差异(P0.05)。结论:本法测定结果的精确度与国标碘量法基本一致,适用于平时及野外特殊条件下饮水消毒中有效氯含量的测定。 相似文献
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针对当前医学教育中,医学教师人文素质水平偏低,在教学过程中偏重专业知识教育而忽视人文素质教育的现状,分析提高医学教师人文素质的必要性,并提出提高医学教师人文素质水平的措施。 相似文献
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本研究利用晕船模拟装置刺激大鼠诱发晕船 ,检测晕船时血浆血管加压素含量 ,为研究舰艇人员的晕船防治提供实验依据。1 材料与方法1 1 实验动物 SD大鼠 2 2只 ,体重为 2 50~ 3 0 0g ,均为雄性。1 2 大鼠晕船模型 利用晕船模拟装置 (按Crampton的报道仿制而成 )诱发大鼠晕船[1]。将 2 2只大鼠无束缚地放入旋转装置的有机玻璃笼内 ,绕水平轴顺时针旋转 ,以 16°/s2 角加速度加速 ,达最大速度 12 0°/s后 ,立即以 48°/s2 的角加速度减速 ,直至旋转停止为一个周期 ,时间 10s ,每天刺激 1h ,旋转刺激一个月。根据实验中大鼠异嗜高岭土行… 相似文献
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目的 探讨脂筏对子宫颈癌细胞生长的影响并初步探讨其作用机制。方法 将HeLa细胞系分为不处理对照组、脂筏干扰剂组及NADPH氧化酶抑制剂组,四甲基偶氮唑蓝(MTT)法测定各组培养24 h后的细胞存活率,Western blot法检测各组细胞内缺氧诱导因子1α(HIF-1α)蛋白相对含量。结果 与对照组相比脂筏干扰剂组细胞的生长速度明显减慢(0.612±0.051 与 0.984±0.034),NADPH氧化酶抑制剂组显示了类似的效应(0.591±0.074与0.984±0.034),差异有统计学意义(t=4.062,P<0.05)。与对照组相比脂筏干扰剂组及NADPH氧化酶抑制剂组HIF-1α的表达量也明显降低(1.79±0.14与 2.56±0.22;1.54±0.12 与 2.56±0.22),差异有统计学意义(t=2.423,P<0.05)。结论 脂筏可能通过NADPH氧化酶激活途径激活HIF-1α及其下游原癌基因促进子宫颈癌细胞的生长,脂筏干扰剂及NADPH氧化酶抑制剂可能成为子宫颈癌药物治疗新的研究方向。 相似文献
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复杂膀胱阴道瘘因瘘孔修复失败率高成为妇科及泌尿外科共同的难题,如何提高手术成功率是医务人员关心的问题。该文从手术时机和手术策略的选择、术中手术要点、影响预后的因素四方面总结经阴道途径的复杂膀胱阴道瘘的治疗经验,为临床工作者提供参考。 相似文献
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BACKGROUND: LL37, the only antimicrobial peptide of the cathelicidin family identified from the human, not only promotes the proliferation of endothelial cells, but also plays an important role in angiogenesis and re-epithelialization.
OBJECTIVE: To construct a recombinant adenovirus overexpressing human antimicrobial peptide LL37 gene, and to detect the expression and secretion of LL37 after transfected into the canine vaginal epithelial cells.
METHODS: The cDNA encoding LL37 was amplified by PCR. Recombinant adenovirus expression plasmid encoding LL37 and green fluorescent protein (EGFP) was constructed, and identified using restriction endonuclease technology and DNA sequencing method. Adenovirus particles were generated by cotransfecting the 293-packaging cell line. The adenovirus were collected, amplified and concentrated, and viral titers were determined by end-point dilution assay by applying serial dilutions of the purified viruses to 293 cells. Primary cultured canine vaginal epithelial cells were transfected by the recombinant adenovirus. The transfection efficacy was observed by fluorescence microscope, and the cultured supernatant was collected to determine the expression of LL37 by ELISA method at 1, 2, 3, 5 and 7 days after transfection.
RESULTS AND CONCLUSION:The adenovirus vector GV314-LL37 with the titer of 3×109 pfu/mL was successfully constructed and identified by DNA sequencing methods. Canine vaginal epithelial cells were successfully isolated and cultured and grew stably. After transfection, vaginal epithelial cells could express the EGFP and LL37 efficiently in a time-dependent manner detected by fluorescence microscope and ELISA method. The transfection efficacy of EGFP reached to 89% at 72 hours. The level of LL37 in the cell culture supernatant in the transfection group was significantly higher than that in the control group, the highest expression of LL37 was found at 3 days that lasting for 7 days. In conclusion, the recombinant adenovirus overexpressing human antimicrobial peptide LL37 gene is successfully constructed, which can express and secrete LL37 after transfected into canine vaginal epithelial cells, providing a foundation for constructing the tissue-engineered vagina possessing anti-infection and neovascularization.
中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程 相似文献