A core-shell Rhodamine B-doped SiO2 nanoparticle was synthesized and its fluorescent intensity was found to be 1000 times higher than that of individual Rhodamine B molecule. The doped nanoparticles were further conju...A core-shell Rhodamine B-doped SiO2 nanoparticle was synthesized and its fluorescent intensity was found to be 1000 times higher than that of individual Rhodamine B molecule. The doped nanoparticles were further conjugated with streptavidin and the resulting nanoparticles were used in the detection of reverse-phase protein microarrays, in which human IgG of various concentrations was first immobilized on aldehyde-modified glass slides and then biotinlyated goat anti human IgG as well as the labeled nanoparticles were sequentially conjugated. The calibration curve is linear over the range from 800 fg to 500 pg and the limit of detection is 100 fg, which is 8 times lower than that of streptavidin-labeled Cy3 fluorescent dyes. The dyedoped SiO2 nanoparticles show potentials for the protein array detection.展开更多
A modified ELISA realizing fluorescent and colorimetric immunoassay of nuclear matrix protein 22 (NMP 22) was developed based on porous Pd nanoparticles. The unique structure and excellent enzyme mimetic activity of p...A modified ELISA realizing fluorescent and colorimetric immunoassay of nuclear matrix protein 22 (NMP 22) was developed based on porous Pd nanoparticles. The unique structure and excellent enzyme mimetic activity of porous Pd nanoparticles favor to oxidize o-phenylenediamine (OPD) into 2,3-phenazinediamine (oxOPD) by H2O2, producing colorimetric and fluorescence dual-readout signal for the detection of NMP 22. The developed immunoassay method will offer great potential in clinical research and diagnostic applications.展开更多
目的追踪骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)在溃疡性结肠炎大鼠模型体内的归巢情况,为BMSCs移植治疗溃疡性结肠炎提供实验基础。方法体外培养扩增Sprague-Dawley(SD)雄性大鼠BMSCs,流式细胞术鉴定BMSCs,采用慢病毒...目的追踪骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)在溃疡性结肠炎大鼠模型体内的归巢情况,为BMSCs移植治疗溃疡性结肠炎提供实验基础。方法体外培养扩增Sprague-Dawley(SD)雄性大鼠BMSCs,流式细胞术鉴定BMSCs,采用慢病毒载体介导的绿色荧光蛋白(green fluorescent protein,GFP)技术(lentivirus-GFP)对BMSCs进行荧光蛋白标记(Ad-GFP-BMSCs)。SD雌性大鼠被随机分为3组:空白组、模型组、Ad-GFP-BMSCs组。2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid,TNBS)诱导溃疡性结肠炎,疾病活动指数(disease activity index,DAI)对各组大鼠进行评估,采用性别交叉移植的方法,尾静脉注射Ad-GFP-BMSCs于雌性大鼠体内,1周后收集结肠标本,对结肠标本进行病理学评估,免疫荧光检测结肠组织GFP表达,PCR检测Y染色体的性别决定区(sex-determining region on the Y chromosome,SRY)基因。结果 TNBS能成功诱导溃疡性结肠炎,与模型组相比,Ad-GFP-BMSCs组DAI评分显著下降(P<0.05)。Lentivirus-GFP成功标记SD大鼠的BMSCs,转染后BMSCs能够稳定表达GFP蛋白,尾静脉注射1周,Ad-GFP-BMSCs组免疫荧光可检测到GFP表达,PCR可检测出SRY基因,空白组和模型组无GFP和SRY基因表达。结论慢病毒载体介导的GFP转染技术和SRY基因可以追踪移植的BMSCs,而且BMSCs通过尾静脉注射可以归巢于受损的结肠组织,这可能为BMSCs移植治疗溃疡性结肠炎提供实验基础。展开更多
基金support from the National Natural Science Foundation of China (Grant Nos. 20575079, 20890020 & 20775033)the National Science Funds for Creative Research Groups (Grant No. 20821063)+1 种基金the National Basic Research Program of China (Grant Nos. 2006CB910803 & 2007CB936404)the Opening Research Foundation of State Key Laboratory of Proteomics
文摘A core-shell Rhodamine B-doped SiO2 nanoparticle was synthesized and its fluorescent intensity was found to be 1000 times higher than that of individual Rhodamine B molecule. The doped nanoparticles were further conjugated with streptavidin and the resulting nanoparticles were used in the detection of reverse-phase protein microarrays, in which human IgG of various concentrations was first immobilized on aldehyde-modified glass slides and then biotinlyated goat anti human IgG as well as the labeled nanoparticles were sequentially conjugated. The calibration curve is linear over the range from 800 fg to 500 pg and the limit of detection is 100 fg, which is 8 times lower than that of streptavidin-labeled Cy3 fluorescent dyes. The dyedoped SiO2 nanoparticles show potentials for the protein array detection.
基金the Natural Science Foundation of Shandong Province(No.ZR2017MB017)
文摘A modified ELISA realizing fluorescent and colorimetric immunoassay of nuclear matrix protein 22 (NMP 22) was developed based on porous Pd nanoparticles. The unique structure and excellent enzyme mimetic activity of porous Pd nanoparticles favor to oxidize o-phenylenediamine (OPD) into 2,3-phenazinediamine (oxOPD) by H2O2, producing colorimetric and fluorescence dual-readout signal for the detection of NMP 22. The developed immunoassay method will offer great potential in clinical research and diagnostic applications.
文摘目的追踪骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)在溃疡性结肠炎大鼠模型体内的归巢情况,为BMSCs移植治疗溃疡性结肠炎提供实验基础。方法体外培养扩增Sprague-Dawley(SD)雄性大鼠BMSCs,流式细胞术鉴定BMSCs,采用慢病毒载体介导的绿色荧光蛋白(green fluorescent protein,GFP)技术(lentivirus-GFP)对BMSCs进行荧光蛋白标记(Ad-GFP-BMSCs)。SD雌性大鼠被随机分为3组:空白组、模型组、Ad-GFP-BMSCs组。2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid,TNBS)诱导溃疡性结肠炎,疾病活动指数(disease activity index,DAI)对各组大鼠进行评估,采用性别交叉移植的方法,尾静脉注射Ad-GFP-BMSCs于雌性大鼠体内,1周后收集结肠标本,对结肠标本进行病理学评估,免疫荧光检测结肠组织GFP表达,PCR检测Y染色体的性别决定区(sex-determining region on the Y chromosome,SRY)基因。结果 TNBS能成功诱导溃疡性结肠炎,与模型组相比,Ad-GFP-BMSCs组DAI评分显著下降(P<0.05)。Lentivirus-GFP成功标记SD大鼠的BMSCs,转染后BMSCs能够稳定表达GFP蛋白,尾静脉注射1周,Ad-GFP-BMSCs组免疫荧光可检测到GFP表达,PCR可检测出SRY基因,空白组和模型组无GFP和SRY基因表达。结论慢病毒载体介导的GFP转染技术和SRY基因可以追踪移植的BMSCs,而且BMSCs通过尾静脉注射可以归巢于受损的结肠组织,这可能为BMSCs移植治疗溃疡性结肠炎提供实验基础。
