目的通过磷脂酰乙醇胺结合蛋白1(recombinant human phosphatidylethanolamine binding protein 1,PEBP1)在先天性肾积水患儿尿液中表达水平的研究,探讨PEBP1作为先天性肾积水的生物学标记物的可行性。方法应用酶联免疫吸附试验(ELISA)...目的通过磷脂酰乙醇胺结合蛋白1(recombinant human phosphatidylethanolamine binding protein 1,PEBP1)在先天性肾积水患儿尿液中表达水平的研究,探讨PEBP1作为先天性肾积水的生物学标记物的可行性。方法应用酶联免疫吸附试验(ELISA)法测定30例重度肾积水患儿、30例轻度肾积水患儿及正常儿童膀胱内尿液样本中PEBP1的表达情况,并将PEBP1的表达水平与肾核素动态显像测定的患肾的分肾功能做相关性分析。结果与正常对照组相比,PEBP1在先天性肾积水患儿尿液中的表达明显下调,积水程度越重,表达越低(P<0.05)。重度肾积水患儿术后PEBP1的表达上调,术后1个月PEBP1的表达仍低于正常对照组(P<0.05)。PEBP1的表达与分肾功能呈正相关性(r=0.818,P<0.05)。当PEBP1≤0.521ng/mgcr.时,患肾的分肾功能下降至30%及以下,接受者操作特征(ROC)曲线下面积0.89,诊断灵敏度80%,特异度90%。结论PEBP1蛋白在先天性肾积水患儿的尿液中的表达明显下调,术后1个月表达上调。PEBP1值越低,患肾的分肾功能越低,肾脏功能损害越重,有较高的诊断灵敏度,有可能成为辅助诊断患儿先天性肾积水的生物学标记物,并可评估病情的严重程度、治疗效果。展开更多
Biomarkers play an important role in the detection at an early stage of pancreatic cancer. The aim of the present study was to optimize the conditions of antibody arrays for detecting Hippocalcin-like 1 （HPCAL1）, ph...Biomarkers play an important role in the detection at an early stage of pancreatic cancer. The aim of the present study was to optimize the conditions of antibody arrays for detecting Hippocalcin-like 1 （HPCAL1）, phosphatidylethanolamine binding protein 1 （PEBP1）, lectin galactoside-binding soluble 7 （LGALS7）, and serpin peptidase inhibitor clade E member 2 （SERPINE2） as biomarkers for pancreatic cancer detection in a single assay and to investigate antibodies’ specificity and cross-reactivity. Capture antibodies against HPCAL1, PEBP1, LGALS7 and SERPINE2 were printed on nitrocellulose coated glass slides. HPCAL1, PEBP1, LGALS7 and SERPINE2 proteins with different concentrations were incubated with the capture antibodies at different temperatures for different time periods. Biotinylated detection antibodies recognizing a different epitope on the captured proteins and a secondary detection molecule （Streptavidin-PE） were used to detect fluorescent signals. The arrays showed the strongest signals when the concentration of the capture antibodies was at 500 μg/mL in PBST0.05 （PBS with 0.05% Tween-20）, and the slides were incubated overnight at 4°C. The lowest protein concentration for detection was 2 ng/mL. Each antibody demonstrated high specificity to the corresponding antigen in detecting a mixture of 4 proteins without significant cross-reactivity. The fluorescence and biomarker concentration displayed a linear correlation. The antibody microarray system could be a useful tool for potential biomarker detection for pancreatic cancer.展开更多
文摘目的通过磷脂酰乙醇胺结合蛋白1(recombinant human phosphatidylethanolamine binding protein 1,PEBP1)在先天性肾积水患儿尿液中表达水平的研究,探讨PEBP1作为先天性肾积水的生物学标记物的可行性。方法应用酶联免疫吸附试验(ELISA)法测定30例重度肾积水患儿、30例轻度肾积水患儿及正常儿童膀胱内尿液样本中PEBP1的表达情况,并将PEBP1的表达水平与肾核素动态显像测定的患肾的分肾功能做相关性分析。结果与正常对照组相比,PEBP1在先天性肾积水患儿尿液中的表达明显下调,积水程度越重,表达越低(P<0.05)。重度肾积水患儿术后PEBP1的表达上调,术后1个月PEBP1的表达仍低于正常对照组(P<0.05)。PEBP1的表达与分肾功能呈正相关性(r=0.818,P<0.05)。当PEBP1≤0.521ng/mgcr.时,患肾的分肾功能下降至30%及以下,接受者操作特征(ROC)曲线下面积0.89,诊断灵敏度80%,特异度90%。结论PEBP1蛋白在先天性肾积水患儿的尿液中的表达明显下调,术后1个月表达上调。PEBP1值越低,患肾的分肾功能越低,肾脏功能损害越重,有较高的诊断灵敏度,有可能成为辅助诊断患儿先天性肾积水的生物学标记物,并可评估病情的严重程度、治疗效果。
基金supported by the Lustgarten Foundation for Pancreatic Cancer Research (No. RFA-08-003)
文摘Biomarkers play an important role in the detection at an early stage of pancreatic cancer. The aim of the present study was to optimize the conditions of antibody arrays for detecting Hippocalcin-like 1 （HPCAL1）, phosphatidylethanolamine binding protein 1 （PEBP1）, lectin galactoside-binding soluble 7 （LGALS7）, and serpin peptidase inhibitor clade E member 2 （SERPINE2） as biomarkers for pancreatic cancer detection in a single assay and to investigate antibodies’ specificity and cross-reactivity. Capture antibodies against HPCAL1, PEBP1, LGALS7 and SERPINE2 were printed on nitrocellulose coated glass slides. HPCAL1, PEBP1, LGALS7 and SERPINE2 proteins with different concentrations were incubated with the capture antibodies at different temperatures for different time periods. Biotinylated detection antibodies recognizing a different epitope on the captured proteins and a secondary detection molecule （Streptavidin-PE） were used to detect fluorescent signals. The arrays showed the strongest signals when the concentration of the capture antibodies was at 500 μg/mL in PBST0.05 （PBS with 0.05% Tween-20）, and the slides were incubated overnight at 4°C. The lowest protein concentration for detection was 2 ng/mL. Each antibody demonstrated high specificity to the corresponding antigen in detecting a mixture of 4 proteins without significant cross-reactivity. The fluorescence and biomarker concentration displayed a linear correlation. The antibody microarray system could be a useful tool for potential biomarker detection for pancreatic cancer.
