目的:分析4例携带血管性血友病因子裂解酶13(a disintegrin and metalloproteinase with thrombospondin type 1 motifs 13,ADAMTS13)去整合素样结构域杂合突变的血栓患者其基因型和表型特征,探讨ADAMTS13功能缺陷与血栓形成之间的关联...目的:分析4例携带血管性血友病因子裂解酶13(a disintegrin and metalloproteinase with thrombospondin type 1 motifs 13,ADAMTS13)去整合素样结构域杂合突变的血栓患者其基因型和表型特征,探讨ADAMTS13功能缺陷与血栓形成之间的关联。方法:通过易栓症基因Panel检测筛查血栓患者的易栓症相关基因突变,纳入4个携带ADAMTS13去整合素样结构域突变的易栓症家系。采用凝固法检测患者的凝血功能,使用免疫比浊法测定血管性血友病因子(von Willebrand factor,vWF)抗原和活性,酶联免疫吸附法检测vWF的胶原结合能力。通过十二烷基硫酸钠-琼脂糖凝胶电泳分析vWF多聚体的分布情况并进行灰度值半定量分析。采用荧光共振能量转移法测定血浆中ADAMTS13活性,酶联免疫吸附法检测ADAMTS13抗原水平。使用PyMOL软件对野生型及突变型ADAMTS13蛋白的三维结构进行对比分析。结果:4个家系的先证者均经历不同程度的血栓事件,包括脑静脉窦血栓、肺栓塞及下肢深静脉血栓。ADAMTS13蛋白去整合素样结构域突变约占ADAMTS13单杂合突变的4/87。遗传分析显示,4例先证者均携带ADAMTS13基因去整合素样结构域单杂合突变(p.Pro301Ala、p.Pro301Arg、p.Arg349Cys),其中p.Pro301Ala和p.Pro301Arg为首次报道突变。凝血功能检测结果表明,4例患者ADAMTS13活性和抗原水平显著降低(Act为57.42%~72.88%,Ag为66.94%~78.34%),vWF活性和抗原水平升高(Act为158.2%~213.7%,Ag为167.2%~216.6%)。vWF多聚体电泳分析显示,患者血浆中高分子量多聚体(high-molecular-weight multimers,HMWMs)比例显著增加(灰度值166.6~218.9比117.4),提示HMWMs较正常人明显增多。结构分析进一步表明,突变位点位于ADAMTS13蛋白去整合素样结构域的关键区域,可能破坏蛋白稳定性及与vWF的结合能力。结论:本研究首次报道了位于ADAMTS13蛋白去整合素样结构域上的2个新突变(Pro301Ala和Pro301Arg),并再次验证了已知突变Arg349Cys的致病特性。结果证实这些突变可导致ADAMTS13蛋白表达水平下降,并进一步引起其酶活性的显著降低,表现为对vWF高分子量多聚体的裂解能力减弱,功能试验证实患者体内存在异常增多的vWF多聚体,破坏了ADAMTS13-vWF轴的动态平衡,从而增加血栓形成的风险。展开更多
目的探讨血浆金属肽酶含血小板反应蛋白13(a disintegrin-like and metalloprotease with thrombospondin type 1 repeats-13,ADAMTS13)水平对心力衰竭合并2型糖尿病(type 2 diabetes mellitus,T2DM)患者的预后价值。方法收集2019年1月...目的探讨血浆金属肽酶含血小板反应蛋白13(a disintegrin-like and metalloprotease with thrombospondin type 1 repeats-13,ADAMTS13)水平对心力衰竭合并2型糖尿病(type 2 diabetes mellitus,T2DM)患者的预后价值。方法收集2019年1月至2020年12月在榆林市第二医院住院的急性失代偿性心力衰竭成年患者临床资料,共纳入278例。采用ELISA试剂盒测定ADAMTS13水平,并基于ADAMTS13三分位数将患者分为T1<9.9μg/L组(n=92),T2≥10.0~18.5μg/L组(n=93)和T3≥18.5μg/L组(n=93)。主要结局定义为全因死亡率或心力衰竭再住院的复合终点。次要结局包括全因死亡率和心力衰竭再住院。结果与ADAMTS13最高三分位数的患者相比,ADAMTS13最低三分位数的患者中T2DM例数、身体质量指数、尿酸、空腹血糖、糖化血红蛋白(glycated hemoglobin,HbA1c)和超敏C反应蛋白(high sensitivity C-reactive protein,hs-CRP)水平均显著增加(P<0.05)。T2DM患者的主要结局、全因死亡和心力衰竭住院的绝对风险高于非T2DM患者(χ^(2)=12.028、8.725、9.023,均P=0.000)。不同ADAMTS13水平患者心力衰竭率的差异有统计学意义(χ^(2)=9.384,P=0.002)。ADAMTS13每增加一倍与合并T2DM的HF患者的主要结局(HR=0.69,95%CI=0.61~0.79)、全因死亡率(HR=0.59,95%CI=0.47-0.74)和心衰再住院(HR=0.75,95%CI=0.65~0.87)下降相关。结论ADAMTS13是HF危险分层的一个独立的生物标志物,特别是在合并T2DM的患者中。展开更多
AIM:To investigate thrombotic microangiopathy (TMA)in liver transplantion,because TMA is an infrequent but life-threatening complication in the transplantation field. METHODS:A total of 206 patients who underwent livi...AIM:To investigate thrombotic microangiopathy (TMA)in liver transplantion,because TMA is an infrequent but life-threatening complication in the transplantation field. METHODS:A total of 206 patients who underwent living-donor liver transplantation (LDLT) were evaluated,and the TMA-like disorder (TMALD) occurred in seven recipients. RESULTS:These TMALD recipients showed poor outcomes in comparison with other 199 recipients. Although two TMALD recipients successfully recovered,the other five recipients finally died despite intensive treatments including repeated plasma exchange (PE) and re-transplantation. Histopathological analysis of liver biopsies after LDLT revealed obvious differences according to the outcomes. Qualitative analysis of antibodies against a disintegrin-like domain and metalloproteinase with thrombospondin type 1 motifs (ADAMTS-13) were negative in all patients. The fragmentation of red cells,the microhemorrhagic macules and the platelet counts were early markers for the suspicion of TMALD after LDLT. Although the absolute values of von Willebrand factor (vWF) and ADAMTS-13 did not necessarily reflect TMALD,the vWF/ADAMTS-13 ratio had a clear diagnostic value in all cases. The establishment of adequate treatments for TMALD,such as PE for ADAMTS-13 replenishment or treatments against inhibitory antibodies,must be decided according to each case. CONCLUSION:The optimal induction of adequate therapies based on early recognition of TMALD by the reliable markers may confer a large advantage for TMALD after LDLT.展开更多
目的:通过系统评价与Meta分析探索Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶7(ADAMTS7)基因rs3825807位点单核苷酸的多态性与冠心病发病风险的关联。方法:计算机检索PubMed, Web of Science, Cochrane Library,中国知网,万方,维普...目的:通过系统评价与Meta分析探索Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶7(ADAMTS7)基因rs3825807位点单核苷酸的多态性与冠心病发病风险的关联。方法:计算机检索PubMed, Web of Science, Cochrane Library,中国知网,万方,维普和中国生物医学数据库,以获取ADAMTS7基因rs3825807多态性与冠心病易感性的原始研究。检索时限均为建库至2019年12月6日。由两位研究者独立筛选文献、提取数据并评价纳入研究的偏倚风险后,采用RevMan 5.3软件进行Meta分析。结果:共纳入6个病例-对照研究,观察组包括4 989例病例,对照组包含5 471例。Meta分析结果显示,患者ADAMTS7基因rs3825807多态性与冠心病的发病风险增加有相关性(AA vs, GG:OR=21.07,95%CI:1.61~275.95,P=0.02;AG vs. GG:OR=6.80,95%CI:0.77~60.11,P=0.08;AA+AG vs. GG:OR=13.19,95%CI:1.09~158.97,P=0.04;AA vs. AG+GG:OR=2.39,95%CI:1.44~3.96,P=0.0007;A vs. G:OR=23.44,95%CI:8.19~67.10,P<0.00001)。结论:患者ADAMTS7基因rs3825807多态性是冠心病的发病风险因素之一。受纳入研究数量和质量限制,本研究需更多的高质量临床研究予以验证。展开更多
目的:检测大鼠蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后早期海马组织中含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-1(a disintegrin-like and metalloproteinase with thrombospondin type l motifs,ADAMTS-1)在基底动脉...目的:检测大鼠蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后早期海马组织中含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-1(a disintegrin-like and metalloproteinase with thrombospondin type l motifs,ADAMTS-1)在基底动脉中的表达,分析其与大鼠SAH后急性脑血管痉挛(cerebral vasospasm,CVS)的相关性,探讨其在SAH后早期脑损伤(earlybrain injury,EBI)中的作用。方法:健康雄性成年SD大鼠108只,体质量250～300g,随机分为SAH组(n=90)和假手术组(sham组,n=18)。SAH组取大鼠自体动脉血,用视交叉池注血法建立大鼠SAH模型(sham组注入等量0.9%氯化钠液)。将SAH组随机分为6h、12h、24h、48h、72h5个亚组,每个亚组18只,分别在相应时间点处死。用蛋白质印迹(Western-blot)方法及实时荧光定量逆转录-聚合酶链反应(RT-PCR)法检测SAH各亚组及sham组基底动脉ADAMTS-1的表达,同时用HE染色法对基底动脉进行形态学观察。探讨ADAMTS-1与实验性大鼠SAH后急性CVS的相关性。结果:与sham组相比,SAH 6h亚组基底动脉ADAMTS-1表达无显著差异;12h亚组基底动脉ADAMTS-1蛋白表达显著增高,24h亚组最高,48h亚组、72h亚组逐渐下降,但仍维持在较高水平。与sham组相比,SAH 6h亚组大鼠基底动脉变化不明显;12h亚组大鼠基底动脉管腔缩小,管壁增厚;24h亚组基底动脉痉挛最为明显;48h亚组基底动脉痉挛较24h亚组减轻;与24h亚组、48h亚组相比,72h亚组基底动脉管腔直径更大、管壁厚度更薄。ADAMTS-1蛋白表达与SAH后急性CVS呈正相关(r=0.916,P=0.003)。结论:大鼠SAH后早期基底动脉ADAMTS-1表达与急性CVS正相关,提示ADAMTS-1可能参与大鼠SAH后EBI的病理过程。展开更多
A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs 13(ADAMTS13) specifically cleaves unusually-large von Willebrand factor(VWF) multimers under high shear stress,and down-regulates VWF function...A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs 13(ADAMTS13) specifically cleaves unusually-large von Willebrand factor(VWF) multimers under high shear stress,and down-regulates VWF function to form platelet thrombi.Deficiency of plasma ADAMTS13 activity induces a life-threatening systemic disease,termed thrombotic microangiopathy(TMA) including thrombotic thrombocytopenic purpura(TTP).Children with advanced biliary cirrhosis due to congenital biliary atresia sometimes showed pathological features of TMA,with a concomitant decrease of plasma ADAMTS13 activity.Disappearance of their clinical findings of TTP after successful liver transplantation suggested that the liver is a major organ producing plasma ADAMTS13.In situ hybridization analysis showed that ADAMTS13 was produced by hepatic stellate cells.Subsequently,it was found that ADADTS13 was not merely responsible to development of TMA and TTP,but also related to some kinds of liver dysfunction after liver transplantation.Ischemia-reperfusion injury and acute rejection in liver transplant recipients were often associated with marked decrease of ADAMTS13 and concomitant formation of unusually large VWF multimers without findings of TMA/TTP.The similar phenomenon was observed also in patients who underwent hepatectomy for liver tumors.Imbalance between ADAMTS13 and VWF in the hepatic sinusoid might cause liver damage due to microcirculatory disturbance.It can be called as "local TTP like mechanism" which plays a crucial role in liver dysfunction after liver transplantation and surgery.展开更多
文摘目的:分析4例携带血管性血友病因子裂解酶13(a disintegrin and metalloproteinase with thrombospondin type 1 motifs 13,ADAMTS13)去整合素样结构域杂合突变的血栓患者其基因型和表型特征,探讨ADAMTS13功能缺陷与血栓形成之间的关联。方法:通过易栓症基因Panel检测筛查血栓患者的易栓症相关基因突变,纳入4个携带ADAMTS13去整合素样结构域突变的易栓症家系。采用凝固法检测患者的凝血功能,使用免疫比浊法测定血管性血友病因子(von Willebrand factor,vWF)抗原和活性,酶联免疫吸附法检测vWF的胶原结合能力。通过十二烷基硫酸钠-琼脂糖凝胶电泳分析vWF多聚体的分布情况并进行灰度值半定量分析。采用荧光共振能量转移法测定血浆中ADAMTS13活性,酶联免疫吸附法检测ADAMTS13抗原水平。使用PyMOL软件对野生型及突变型ADAMTS13蛋白的三维结构进行对比分析。结果:4个家系的先证者均经历不同程度的血栓事件,包括脑静脉窦血栓、肺栓塞及下肢深静脉血栓。ADAMTS13蛋白去整合素样结构域突变约占ADAMTS13单杂合突变的4/87。遗传分析显示,4例先证者均携带ADAMTS13基因去整合素样结构域单杂合突变(p.Pro301Ala、p.Pro301Arg、p.Arg349Cys),其中p.Pro301Ala和p.Pro301Arg为首次报道突变。凝血功能检测结果表明,4例患者ADAMTS13活性和抗原水平显著降低(Act为57.42%~72.88%,Ag为66.94%~78.34%),vWF活性和抗原水平升高(Act为158.2%~213.7%,Ag为167.2%~216.6%)。vWF多聚体电泳分析显示,患者血浆中高分子量多聚体(high-molecular-weight multimers,HMWMs)比例显著增加(灰度值166.6~218.9比117.4),提示HMWMs较正常人明显增多。结构分析进一步表明,突变位点位于ADAMTS13蛋白去整合素样结构域的关键区域,可能破坏蛋白稳定性及与vWF的结合能力。结论:本研究首次报道了位于ADAMTS13蛋白去整合素样结构域上的2个新突变(Pro301Ala和Pro301Arg),并再次验证了已知突变Arg349Cys的致病特性。结果证实这些突变可导致ADAMTS13蛋白表达水平下降,并进一步引起其酶活性的显著降低,表现为对vWF高分子量多聚体的裂解能力减弱,功能试验证实患者体内存在异常增多的vWF多聚体,破坏了ADAMTS13-vWF轴的动态平衡,从而增加血栓形成的风险。
基金Supported by the Grant from Uehara Memorial Foundation, No. 200940051, Tokyo, 171-0033, Japan
文摘AIM:To investigate thrombotic microangiopathy (TMA)in liver transplantion,because TMA is an infrequent but life-threatening complication in the transplantation field. METHODS:A total of 206 patients who underwent living-donor liver transplantation (LDLT) were evaluated,and the TMA-like disorder (TMALD) occurred in seven recipients. RESULTS:These TMALD recipients showed poor outcomes in comparison with other 199 recipients. Although two TMALD recipients successfully recovered,the other five recipients finally died despite intensive treatments including repeated plasma exchange (PE) and re-transplantation. Histopathological analysis of liver biopsies after LDLT revealed obvious differences according to the outcomes. Qualitative analysis of antibodies against a disintegrin-like domain and metalloproteinase with thrombospondin type 1 motifs (ADAMTS-13) were negative in all patients. The fragmentation of red cells,the microhemorrhagic macules and the platelet counts were early markers for the suspicion of TMALD after LDLT. Although the absolute values of von Willebrand factor (vWF) and ADAMTS-13 did not necessarily reflect TMALD,the vWF/ADAMTS-13 ratio had a clear diagnostic value in all cases. The establishment of adequate treatments for TMALD,such as PE for ADAMTS-13 replenishment or treatments against inhibitory antibodies,must be decided according to each case. CONCLUSION:The optimal induction of adequate therapies based on early recognition of TMALD by the reliable markers may confer a large advantage for TMALD after LDLT.
文摘目的:通过系统评价与Meta分析探索Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶7(ADAMTS7)基因rs3825807位点单核苷酸的多态性与冠心病发病风险的关联。方法:计算机检索PubMed, Web of Science, Cochrane Library,中国知网,万方,维普和中国生物医学数据库,以获取ADAMTS7基因rs3825807多态性与冠心病易感性的原始研究。检索时限均为建库至2019年12月6日。由两位研究者独立筛选文献、提取数据并评价纳入研究的偏倚风险后,采用RevMan 5.3软件进行Meta分析。结果:共纳入6个病例-对照研究,观察组包括4 989例病例,对照组包含5 471例。Meta分析结果显示,患者ADAMTS7基因rs3825807多态性与冠心病的发病风险增加有相关性(AA vs, GG:OR=21.07,95%CI:1.61~275.95,P=0.02;AG vs. GG:OR=6.80,95%CI:0.77~60.11,P=0.08;AA+AG vs. GG:OR=13.19,95%CI:1.09~158.97,P=0.04;AA vs. AG+GG:OR=2.39,95%CI:1.44~3.96,P=0.0007;A vs. G:OR=23.44,95%CI:8.19~67.10,P<0.00001)。结论:患者ADAMTS7基因rs3825807多态性是冠心病的发病风险因素之一。受纳入研究数量和质量限制,本研究需更多的高质量临床研究予以验证。
文摘目的:检测大鼠蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后早期海马组织中含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-1(a disintegrin-like and metalloproteinase with thrombospondin type l motifs,ADAMTS-1)在基底动脉中的表达,分析其与大鼠SAH后急性脑血管痉挛(cerebral vasospasm,CVS)的相关性,探讨其在SAH后早期脑损伤(earlybrain injury,EBI)中的作用。方法:健康雄性成年SD大鼠108只,体质量250～300g,随机分为SAH组(n=90)和假手术组(sham组,n=18)。SAH组取大鼠自体动脉血,用视交叉池注血法建立大鼠SAH模型(sham组注入等量0.9%氯化钠液)。将SAH组随机分为6h、12h、24h、48h、72h5个亚组,每个亚组18只,分别在相应时间点处死。用蛋白质印迹(Western-blot)方法及实时荧光定量逆转录-聚合酶链反应(RT-PCR)法检测SAH各亚组及sham组基底动脉ADAMTS-1的表达,同时用HE染色法对基底动脉进行形态学观察。探讨ADAMTS-1与实验性大鼠SAH后急性CVS的相关性。结果:与sham组相比,SAH 6h亚组基底动脉ADAMTS-1表达无显著差异;12h亚组基底动脉ADAMTS-1蛋白表达显著增高,24h亚组最高,48h亚组、72h亚组逐渐下降,但仍维持在较高水平。与sham组相比,SAH 6h亚组大鼠基底动脉变化不明显;12h亚组大鼠基底动脉管腔缩小,管壁增厚;24h亚组基底动脉痉挛最为明显;48h亚组基底动脉痉挛较24h亚组减轻;与24h亚组、48h亚组相比,72h亚组基底动脉管腔直径更大、管壁厚度更薄。ADAMTS-1蛋白表达与SAH后急性CVS呈正相关(r=0.916,P=0.003)。结论:大鼠SAH后早期基底动脉ADAMTS-1表达与急性CVS正相关,提示ADAMTS-1可能参与大鼠SAH后EBI的病理过程。
文摘A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs 13(ADAMTS13) specifically cleaves unusually-large von Willebrand factor(VWF) multimers under high shear stress,and down-regulates VWF function to form platelet thrombi.Deficiency of plasma ADAMTS13 activity induces a life-threatening systemic disease,termed thrombotic microangiopathy(TMA) including thrombotic thrombocytopenic purpura(TTP).Children with advanced biliary cirrhosis due to congenital biliary atresia sometimes showed pathological features of TMA,with a concomitant decrease of plasma ADAMTS13 activity.Disappearance of their clinical findings of TTP after successful liver transplantation suggested that the liver is a major organ producing plasma ADAMTS13.In situ hybridization analysis showed that ADAMTS13 was produced by hepatic stellate cells.Subsequently,it was found that ADADTS13 was not merely responsible to development of TMA and TTP,but also related to some kinds of liver dysfunction after liver transplantation.Ischemia-reperfusion injury and acute rejection in liver transplant recipients were often associated with marked decrease of ADAMTS13 and concomitant formation of unusually large VWF multimers without findings of TMA/TTP.The similar phenomenon was observed also in patients who underwent hepatectomy for liver tumors.Imbalance between ADAMTS13 and VWF in the hepatic sinusoid might cause liver damage due to microcirculatory disturbance.It can be called as "local TTP like mechanism" which plays a crucial role in liver dysfunction after liver transplantation and surgery.
