Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR lev...Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR level with prognosis of MM.Methods u PAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow(control) were examined by ELISA.Furthermore,u PAR expression in bone marrow tissue was investigated by RT-PCR and Western blot,respectively.The distribution of u PAR in MM cells was examined using immunofluorescence staining.The pathological changes in different stages of MM patients were studied by HE staining.Results u PAR level in bone marrow fluid of MM patients(1.52±0.32 μg/ml) was found to be higher than that in the control group(0.98±0.15 μg/ml).Interestingly,u PAR protein(0.686±0.075 vs.0.372±0.043,P<0.05) and m RNA(2.51±0.46 vs.4.46±1.15,P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage.The expression of u PAR in MM bone marrow was confirmed by immunofluorescence staining.Moreover,HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma.Conclusion Our study reveals that u PAR expression is positively correlated with the development and progress of MM.展开更多
目的建立SYBR Green I染料法荧光RT-PCR方法,快速鉴别呼吸道合胞病毒A和B亚型,实现准确、简便、低成本分型检测。方法比对175株呼吸道合胞病毒全基因序列,锁定N基因设计特异性引物,用SYBR Green I染料法荧光PT-PCR扩增A和B亚型呼吸道合...目的建立SYBR Green I染料法荧光RT-PCR方法,快速鉴别呼吸道合胞病毒A和B亚型,实现准确、简便、低成本分型检测。方法比对175株呼吸道合胞病毒全基因序列,锁定N基因设计特异性引物,用SYBR Green I染料法荧光PT-PCR扩增A和B亚型呼吸道合胞病毒,通过熔解曲线分析,测定A、B亚型扩增产物的熔解温度;进行特异性、重复性和准确性评价。结果熔解曲线分析显示,呼吸道合胞病毒A型和B亚型的扩增产物熔解温度相差较大,A亚型为(80.76±0.10)℃,B亚型为(79.00±0.09)℃。特异性试验表明,该方法能检测呼吸道合胞病毒A和B亚型,与甲/乙型流感病毒、新冠病毒、鼻病毒、副流感病毒、博卡病毒、偏肺病毒等呼吸道病毒没有交叉反应。重复性试验表明,该方法扩增呼吸道合胞病毒A和B亚型熔解温度变异系数均低于1%,重复性良好。对56份呼吸道合胞病毒核酸样本的检测结果与商品化TaqMan探针法分型试剂盒完全一致。结论建立的SYBR Green I染料法荧光RT-PCR方法可快速检测呼吸道合胞病毒并鉴别A和B亚型。展开更多
基金Supported by Huzhou Science and Technology Bureau and Huzhou Central Hospital(2014GYB14)
文摘Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR level with prognosis of MM.Methods u PAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow(control) were examined by ELISA.Furthermore,u PAR expression in bone marrow tissue was investigated by RT-PCR and Western blot,respectively.The distribution of u PAR in MM cells was examined using immunofluorescence staining.The pathological changes in different stages of MM patients were studied by HE staining.Results u PAR level in bone marrow fluid of MM patients(1.52±0.32 μg/ml) was found to be higher than that in the control group(0.98±0.15 μg/ml).Interestingly,u PAR protein(0.686±0.075 vs.0.372±0.043,P<0.05) and m RNA(2.51±0.46 vs.4.46±1.15,P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage.The expression of u PAR in MM bone marrow was confirmed by immunofluorescence staining.Moreover,HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma.Conclusion Our study reveals that u PAR expression is positively correlated with the development and progress of MM.
文摘目的建立SYBR Green I染料法荧光RT-PCR方法,快速鉴别呼吸道合胞病毒A和B亚型,实现准确、简便、低成本分型检测。方法比对175株呼吸道合胞病毒全基因序列,锁定N基因设计特异性引物,用SYBR Green I染料法荧光PT-PCR扩增A和B亚型呼吸道合胞病毒,通过熔解曲线分析,测定A、B亚型扩增产物的熔解温度;进行特异性、重复性和准确性评价。结果熔解曲线分析显示,呼吸道合胞病毒A型和B亚型的扩增产物熔解温度相差较大,A亚型为(80.76±0.10)℃,B亚型为(79.00±0.09)℃。特异性试验表明,该方法能检测呼吸道合胞病毒A和B亚型,与甲/乙型流感病毒、新冠病毒、鼻病毒、副流感病毒、博卡病毒、偏肺病毒等呼吸道病毒没有交叉反应。重复性试验表明,该方法扩增呼吸道合胞病毒A和B亚型熔解温度变异系数均低于1%,重复性良好。对56份呼吸道合胞病毒核酸样本的检测结果与商品化TaqMan探针法分型试剂盒完全一致。结论建立的SYBR Green I染料法荧光RT-PCR方法可快速检测呼吸道合胞病毒并鉴别A和B亚型。
