目的探讨血清铁调节蛋白2(iron regulatory protein 2,IRP2)、细胞周期蛋白依赖性激酶抑制剂1A(cyclin-dependent kinase inhibitor 1A,CDKN1A)、人Ⅱ型肺泡细胞表面抗原(human alveolar typeⅡcell surface antigen,KL-6)水平与慢性阻...目的探讨血清铁调节蛋白2(iron regulatory protein 2,IRP2)、细胞周期蛋白依赖性激酶抑制剂1A(cyclin-dependent kinase inhibitor 1A,CDKN1A)、人Ⅱ型肺泡细胞表面抗原(human alveolar typeⅡcell surface antigen,KL-6)水平与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者肺功能及临床预后的相关性。方法回顾性选取136例COPD患者为观察组,选取同期健康体检者136例为对照组。比较两组及不同预后患者血清IRP2、CDKN1A、KL6水平,并比较两组肺功能指标[第1秒用力呼气容积(forced expiratory volume in the first second,FEV_(1))与用力肺活量(forced vital capacity,FVC)的比值(FEV_(1)/FVC)、FEV_(1)占预计值百分比(forced expiratory volume in the first second as a percentage of the predicted value,FEV_(1)%_(pred))]。分析血清IRP2、CDKN1A、KL-6水平与FEV_(1)%_(pred)、FEV_(1)/FVC及预后的相关性,并分析各血清指标联合检测的预测价值。结果观察组血清IRP2、CDKN1A、KL-6水平明显高于对照组(均P<0.01);观察组FEV_(1)%_(pred)、FEV_(1)/FVC明显低于对照组(均P<0.01)。预后不良患者血清IRP2、CDKN1A、KL-6水平明显高于预后良好患者(均P<0.01)。血清IRP2、CDKN1A、KL-6水平与FEV_(1)%_(pred)、FEV_(1)/FVC呈负相关,与患者预后呈正相关(均P<0.01)。血清IRP2、CDKN1A、KL-6水平联合预测COPD患者预后不良的AUC值为0.937,约登指数为0.775,敏感性、特异性分别为94.12%、83.33%,且差异有统计学意义(均P<0.01)。结论血清IRP2、CDKN1A、KL-6水平随着COPD患者肺功能及临床预后的不同而改变,且存在较强相关性。各指标联合检测可为临床预测患者预后提供一定参考。展开更多
为了解临床禽源致病性大肠杆菌中高致病性毒力岛(High pathogenicity island HPI)流行情况和基因特征,根据Gen Bank已知禽源irp2基因序列,设计、合成一对特异性引物,建立irp2基因PCR检测方法,优化、确定PCR扩增特性,进行敏感性、特异性...为了解临床禽源致病性大肠杆菌中高致病性毒力岛(High pathogenicity island HPI)流行情况和基因特征,根据Gen Bank已知禽源irp2基因序列,设计、合成一对特异性引物,建立irp2基因PCR检测方法,优化、确定PCR扩增特性,进行敏感性、特异性、重复性检测评价。对256份临床分离禽源大肠杆菌进行irp2基因PCR检测和基因遗传变异分析。结果显示,建立的PCR检测方法敏感性可达2.14×10^(-4)ng/μL;特异性显示与鸡沙门菌、副鸡嗜血杆菌、鸭疫里默菌、禽巴氏杆菌、鸡毒支原体、鸡新城疫病毒、禽流感病毒(H9N5)核酸均无交叉反应;重复性显示3份阳性样品重复5次检测,变异系数3.4%~5.0%。256份临床样品PCR检测irp2基因,阳性率30.6%。获得了9株分离株irp2基因序列,分析显示,与GenBank已知基因同源性高达96.2%以上。说明建立的禽源大肠杆菌HPI毒力岛irp2基因PCR检测方法具有良好的特异性、敏感性和重复性,可应用于临床致病性大肠杆菌毒力岛基因快速检测。展开更多
Iron accumulation in the brain is associated with the pathogenesis of Parkinson's disease(PD),Misexpression of some iron transport and storage proteins is related to iron dyshorneostasis.Iron regulatory proteins(I...Iron accumulation in the brain is associated with the pathogenesis of Parkinson's disease(PD),Misexpression of some iron transport and storage proteins is related to iron dyshorneostasis.Iron regulatory proteins(IRPs)including IRP1 and IRP2 are cytosolic proteins that play important roles in maintaining cellular iron homeostasis.F-box and leucine-rich repeat protein 5(FBXL5)is involved in the regulation of iron metabolism by degrading IRP2 through the ubiquitin-proteasome system.Nitric oxide(NO)enhances the binding activity of IRP 1,but its effect on IRP2 is ambiguous.Therefore,in the present study,we aim to determine whether sodium nitroprusside(SNP),a NO donor,regulates FBXL5 and IRP2 expression in cultured SH-SY5Y cells.MTT assay revealed that treat-ment of SNP attenuated the cell viability in a dose-dependent manner.Flow cytometry test showed that 100 and 300 ttmol/L SNP administration significantly reduced the mitochondrial membrane potential by 45%and 60%,respectively.Moreover,Western blotting analysis demonstrated that 300 txmol/L SNP significantly increased FBXL5 expression by about 39%,whereas the expression of IRP2 was decreased by 46%,correspondingly.These findings provide evidence that SNP could induce mitochondrial dysfunction,enhance FBXL5 expression and decrease IRP2 expression in SH-SY5Y cells.展开更多
文摘目的探讨血清铁调节蛋白2(iron regulatory protein 2,IRP2)、细胞周期蛋白依赖性激酶抑制剂1A(cyclin-dependent kinase inhibitor 1A,CDKN1A)、人Ⅱ型肺泡细胞表面抗原(human alveolar typeⅡcell surface antigen,KL-6)水平与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者肺功能及临床预后的相关性。方法回顾性选取136例COPD患者为观察组,选取同期健康体检者136例为对照组。比较两组及不同预后患者血清IRP2、CDKN1A、KL6水平,并比较两组肺功能指标[第1秒用力呼气容积(forced expiratory volume in the first second,FEV_(1))与用力肺活量(forced vital capacity,FVC)的比值(FEV_(1)/FVC)、FEV_(1)占预计值百分比(forced expiratory volume in the first second as a percentage of the predicted value,FEV_(1)%_(pred))]。分析血清IRP2、CDKN1A、KL-6水平与FEV_(1)%_(pred)、FEV_(1)/FVC及预后的相关性,并分析各血清指标联合检测的预测价值。结果观察组血清IRP2、CDKN1A、KL-6水平明显高于对照组(均P<0.01);观察组FEV_(1)%_(pred)、FEV_(1)/FVC明显低于对照组(均P<0.01)。预后不良患者血清IRP2、CDKN1A、KL-6水平明显高于预后良好患者(均P<0.01)。血清IRP2、CDKN1A、KL-6水平与FEV_(1)%_(pred)、FEV_(1)/FVC呈负相关,与患者预后呈正相关(均P<0.01)。血清IRP2、CDKN1A、KL-6水平联合预测COPD患者预后不良的AUC值为0.937,约登指数为0.775,敏感性、特异性分别为94.12%、83.33%,且差异有统计学意义(均P<0.01)。结论血清IRP2、CDKN1A、KL-6水平随着COPD患者肺功能及临床预后的不同而改变,且存在较强相关性。各指标联合检测可为临床预测患者预后提供一定参考。
文摘为了解临床禽源致病性大肠杆菌中高致病性毒力岛(High pathogenicity island HPI)流行情况和基因特征,根据Gen Bank已知禽源irp2基因序列,设计、合成一对特异性引物,建立irp2基因PCR检测方法,优化、确定PCR扩增特性,进行敏感性、特异性、重复性检测评价。对256份临床分离禽源大肠杆菌进行irp2基因PCR检测和基因遗传变异分析。结果显示,建立的PCR检测方法敏感性可达2.14×10^(-4)ng/μL;特异性显示与鸡沙门菌、副鸡嗜血杆菌、鸭疫里默菌、禽巴氏杆菌、鸡毒支原体、鸡新城疫病毒、禽流感病毒(H9N5)核酸均无交叉反应;重复性显示3份阳性样品重复5次检测,变异系数3.4%~5.0%。256份临床样品PCR检测irp2基因,阳性率30.6%。获得了9株分离株irp2基因序列,分析显示,与GenBank已知基因同源性高达96.2%以上。说明建立的禽源大肠杆菌HPI毒力岛irp2基因PCR检测方法具有良好的特异性、敏感性和重复性,可应用于临床致病性大肠杆菌毒力岛基因快速检测。
文摘【目的】本研究探讨了云南撒坝猪EPEC HPI毒力岛的摄铁功能与致病性之间的关系。【方法】本试验通过摄铁实验对5株肠致病性E.coli(A-E)和CVCC1565铁载体的合成和在不同2,2’-联吡啶含量下的最低生长抑制浓度及对数期生长速率进行了测定,同时利用已构建的E.coli irp2基因缺失株和亲本株进行对比,检测HMWP2蛋白的表达,并通过半数致死量实验比较两者之间的毒力。【结果】除E.coli(D)外,其余菌株的2,2’-联吡啶最低生长抑制浓度为0.6 m M;亲本株能够表达HMWP2蛋白,irp2缺失株不表达该蛋白;CAS固体培养基中菌株能产生橘黄色的透明晕圈;当2,2’-联吡啶浓度高于0.2 m M时,亲本株的生长率高于irp2缺失株;亲本株的LD50(8.73×10~7cfu/m L)低于缺失株(9.95×10~7cfu/m L)。【结论】云南撒坝猪EPEC HPI铁摄取功能和致病性之间存在着一定关系。
基金supported by the National Natural Science Foundation of China(No.31471114,31500837 and 31540075)Taishan Scholarship,the Key Research and Development Program of Shandong Province,China(No.2016GSF201053)+1 种基金Qingdao Municipal Science and Technology Project(No.16-6-2-2-nsh)the Shandong Provincial Natural Science Foundation of China(No.BS2015SW022)
文摘Iron accumulation in the brain is associated with the pathogenesis of Parkinson's disease(PD),Misexpression of some iron transport and storage proteins is related to iron dyshorneostasis.Iron regulatory proteins(IRPs)including IRP1 and IRP2 are cytosolic proteins that play important roles in maintaining cellular iron homeostasis.F-box and leucine-rich repeat protein 5(FBXL5)is involved in the regulation of iron metabolism by degrading IRP2 through the ubiquitin-proteasome system.Nitric oxide(NO)enhances the binding activity of IRP 1,but its effect on IRP2 is ambiguous.Therefore,in the present study,we aim to determine whether sodium nitroprusside(SNP),a NO donor,regulates FBXL5 and IRP2 expression in cultured SH-SY5Y cells.MTT assay revealed that treat-ment of SNP attenuated the cell viability in a dose-dependent manner.Flow cytometry test showed that 100 and 300 ttmol/L SNP administration significantly reduced the mitochondrial membrane potential by 45%and 60%,respectively.Moreover,Western blotting analysis demonstrated that 300 txmol/L SNP significantly increased FBXL5 expression by about 39%,whereas the expression of IRP2 was decreased by 46%,correspondingly.These findings provide evidence that SNP could induce mitochondrial dysfunction,enhance FBXL5 expression and decrease IRP2 expression in SH-SY5Y cells.
