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  • 作者:

    Objective Nephrolithiasis is one of the most common disorders of the urinary tract. The aim of this study was to examine a possible relationship between DNase I/II activity and E3 SUMO-protein ligase NSE2 in the sera of nephrolithiasis patients to evaluate the possibility of a new biomarker for evaluating kidney damage.
    Methods Sixty nephrolithiasis patients and 50 control patients were enrolled in a case-control study. Their blood urea, creatinine, protein levels and DNase I/II activity levels were measured by spectrometry. Serum NSMCE2 levels were measured by ELISA. Blood was collected from patients of the government health clinics in Kuantan-Pahang and fulfilled the inclusion criteria.
    Results The result indicated that mean levels of sera NSMCE2 have a significantly increase (P<0.01) in patients compared to control group. Compared with control subjects, activities and specific activities of serum DNase I and II were significantly elevated in nephrolithiasis patients (P<0.01).
    Conclusion This study suggests that an increase in serum concentrations of DNase I/II and E3 SUMO-protein ligase NSE2 level can be used as indicators for the diagnosis of kidney injury in patients with nephrolithiasis.

  • 人胚胎滋养细胞和胎盘间充质干细胞的分离与纯化

    作者:沙文琼;王自能;王冬菊

    背景:胎盘的细胞成分较复杂,其中所包含的滋养细胞在母胎免疫耐受过程中起重要作用,胎盘间充质干细胞具有多向分化潜能以及抑制淋巴细胞增殖的特性,常规分离方法通常难以获得大量且纯度较高的上述两种细胞.目的:拟建立一次操作即可同时获得大量、较高纯度的滋养细胞和胎盘间充质于细胞的操作方案.方法:人胎盘组织洗净剪碎,采用胰蛋白酶和DNAse I共同消化,分3个阶段,每个阶段在恒温37℃下180 r/min消化2 min.重悬消化产物,200目筛网过滤后采用Percoll密度梯度分离液分离,分别收集滋养细胞层和胎盘间充质干细胞层.滋养细胞层以差速贴壁法去除成纤维细胞,胎盘间充质干细胞直接接种于75 cm2培养瓶中培养.观察胎盘组织消化情况,计数滋养细胞数量及其细胞角蛋白7的表达,观察胎盘间充质干细胞生长增殖、表型及成骨分化潜能.结果与结论:经胰蛋白酶和DNAse I消化后,胎盘组织仅剩少许残渣.差速贴壁后,滋养细胞数达(5.48±1.98)×10~8个,细胞角蛋白7阳性率为(90+4.36)%.胎盘间充质干细胞接种19~21 d达90%融合,细胞数为(1.96±0.24)×10~6个,强表达CD29,CD44和HLA-ABC,不表达CD34,CD45,CD14和HLA-DR,诱导后茜素红染色呈鲜艳橙红色,可向成骨细胞方向分化.提示采用胰蛋白酶和DNAse I共同消化胎盘组织,并结合Percoll不连续密度梯度分离液分离细胞,可同时一次性获得大量的滋养细胞和较多的胎盘间充质干细胞,细胞纯度和活性均较好.

  • DNaseⅠ对金黄色葡萄球菌生物膜形成的影响

    作者:徐巧玲;孙凤军;冯伟;刘晓;刘亚巍

    目的:研究DNase I对金黄色葡萄球菌生物膜形成的影响。方法采用分光光度计法检测金黄色葡萄球菌的生长曲线,菌落计数法分析金黄色葡萄球菌的粘附能力,96孔板结晶紫染色法检测金黄色葡萄球菌生物膜的形成。结果不同浓度DNase I对金黄色葡萄球菌的生长无影响,然而能显著抑制细菌生物膜的形成,并呈浓度依赖性。此外,DNase I能抑制不同生长期金黄色葡萄球菌的粘附性。与抗菌药物和DNase I单独应用相比,DNase I和抗菌药物联合应用更能显著抑制和分解金黄色葡萄球菌成熟生物膜。结论 DNase I可以有效抑制金黄色葡萄球菌生物膜的形成,并能增强抗菌药物对金黄色葡萄球菌生物膜的抑制作用。

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