目的分析血清血管性血友病因子特异性裂解酶(a disintegrin-like and metalloprotease with thrombospondin type1 motif,member 13,ADAMTS13)水平与接受体外膜肺氧合(extracorporeal membrane oxygenation,ECMO)治疗患者的预后关系。...目的分析血清血管性血友病因子特异性裂解酶(a disintegrin-like and metalloprotease with thrombospondin type1 motif,member 13,ADAMTS13)水平与接受体外膜肺氧合(extracorporeal membrane oxygenation,ECMO)治疗患者的预后关系。方法本研究选取2020年1月—2022年6月在湖南医药学院第一附属医院接受ECMO治疗的42例患者,按照ECMO撤离1个月内预后分为生存组(n=29)和死亡组(n=13)。通过分析接受ECMO治疗患者的血清ADAMTS13水平,分析血清ADAMTS13与接受ECMO治疗患者的预后。结果生存组年龄小于死亡组(P<0.001)。2组性别和体表面积比较,差异无统计学意义(P>0.05)。生存组平均动脉压为(54.24±5.81)mmHg、氧合指数为(178.84±29.38)mmHg,均高于死亡组的(52.56±4.42)mmHg、(155.71±23.12)mmHg;生存组急性生理与慢性健康评分(acute physiology and chronic health evaluation-Ⅱ,APACHE-Ⅱ)评分为(22.51±2.62)分,低于生存组的(27.35±3.85)分(P<0.05)。2组左室射血分数和ECMO流量比较,差异无统计学意义(P>0.05)。ECMO治疗后,2组血清ADAMTS13水平升高。生存组ECMO治疗前1 h及治疗后24 h、48 h血清ADAMTS13水平高于死亡组,差异有统计学意义(P<0.001)。ECMO治疗48 h后血清ADAMTS13水平为ECMO辅助治疗患者死亡的独立危险因素(P<0.001)。结论接受ECMO治疗患者的血清ADAMTS13水平降低与预后不佳相关。展开更多
目的本研究旨在研究和探讨血管性血友病因子（von Willebrand factor，vwF）和含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶（A disintegrin—like and metalloprotease with thrombospondin type 1 motif，vWF裂解酶，ADAMTS13）...目的本研究旨在研究和探讨血管性血友病因子（von Willebrand factor，vwF）和含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶（A disintegrin—like and metalloprotease with thrombospondin type 1 motif，vWF裂解酶，ADAMTS13）的活性在感染性休克和脓毒症患者早期及预后评估方面的临床意义。方法选取2012—06—2013—06收住无锡市人民医院ICU的感染性休克或脓毒症患者40例，根据预后分为存活组20例和死亡组20例，记录其一般情况及入组时APACHE11评分，于发病第1、3、5、7天分别运用ELISA法检测其血浆vWF活性及ADAMTS13活性，各样本均数用方差分析、受试者工作特征（receiver operating characteristic，ROC）曲线分析、参数间进行Spearman相关分析，并绘制ROC曲线探讨vWF活性、ADAMTS13活性与APACHE Ⅱ评分之间的相关性。结果死亡组的vWF活性均明显高于存活组（P〈0．05），ADAMTS13活性明显低于存活组（P〈0．05）；同时死亡组APACHEⅡ评分明显高于存活组（P〈0．05）；存活组在第3天vwF活性升高、ADAMTS13活性下降、APACHEⅡ评分升高，而在第5天和第7天又出现vWF活性持续下降、ADAMTS13活性持续升高、APACHEⅡ评分持续下降，死亡组则呈现vWF活性持续上升、ADAMTS13活性持续下降、APACHEⅡ评分持续升高（P〈0．05）。40例患者的平均vWF活性与APACHEⅡ评分呈正相关（P〈0．01），而ADAMTS13活性与APACHEⅡ评分呈负相关（P〈0．01）：ROC曲线分析，vWF活性和ADAMTS13活性AUC均接近于1，提示能很好地预测患者的预后（P〈0．01）。结论vWF活性、ADAMTS13活性均是判断感染性休克和脓毒症患者预后的可靠指标。在脓毒症的病理过程中，此二者数值的变化与疾病严重程度密切相关，即vWF活性的升高、ADAMTS13活性的降低预示了患者预后差。展开更多
微循环障碍参与了多种疾病的进展,例如脓毒症、多器官衰竭等。血管性假性血友病因子（von Willebrand factor,vWF）可使血小板粘附聚集于血管内皮细胞,导致血小板血栓形成;血管性血友病因子特异性裂解酶（a disintegrin-like and metall...微循环障碍参与了多种疾病的进展,例如脓毒症、多器官衰竭等。血管性假性血友病因子（von Willebrand factor,vWF）可使血小板粘附聚集于血管内皮细胞,导致血小板血栓形成;血管性血友病因子特异性裂解酶（a disintegrin-like and metalloprotease with thrombo-spondin type I repeats 13,ADAMTSl3）可裂解vWF多聚体，预防血栓形成.展开更多
背景:血管性血友病因子水解蛋白酶13(a disintegrin and metalloproteinase with a thrombospondin type 1motif,member 13,ADAMTS13)可结合并酶切血管性血友病因子,在防止血小板过多聚集和血栓的形成中起到关键作用。重组ADAMTS13通常...背景:血管性血友病因子水解蛋白酶13(a disintegrin and metalloproteinase with a thrombospondin type 1motif,member 13,ADAMTS13)可结合并酶切血管性血友病因子,在防止血小板过多聚集和血栓的形成中起到关键作用。重组ADAMTS13通常选择在真核细胞中表达分泌,然而对于其表达纯化目前仍缺乏一个明确高效的途径。目的:寻找合适的宿主细胞,得到稳定表达野生型ADAMTS13和功能增强型ADAMTS13的细胞株,建立高纯度ADAMTS13的纯化方法,进而研究其生物学功能。方法:以野生型ADAMTS13重组质粒为模板,利用位点突变试剂盒得到功能增强型ADAMTS13重组质粒。比较CHO-S细胞、CHO-K1细胞、293T细胞对ADAMTS13的表达效果,挑选出合适的宿主细胞。通过Ni柱亲和层析和分子筛纯化蛋白;采用7.5%SDS-PAGE、Western-blot鉴定野生型ADAMTS13和功能增强型ADAMTS13的纯度及特异性;利用原子力显微镜技术检测野生型及功能增强型ADAMTS13与血管性血友病因子A2的相互作用,鉴定两种蛋白的活性。结果与结论:相对于CHO-S和CHO-K1,293T细胞更适合作为野生型ADAMTS13和功能增强型ADAMTS13的表达宿主;经Ni柱亲和层析和分子筛纯化后,所得到的野生型ADAMTS13和功能增强型ADAMTS13质量浓度分别为103.7,149.7 mg/L,且两种蛋白纯度均约90%;原子力显微镜检测结果显示,野生型ADAMTS13、功能增强型ADAMTS13与血管性血友病因子A2的黏附频率分别为11.37%,14.70%,表明功能增强型ADAMTS13与血管性血友病因子A2的结合亲和力更高,这与近期ADAMTS13构象研究一致。展开更多
含I型血小板结合蛋白基序的解聚蛋白样金属蛋白酶（a disintegrin-like and metalloprotease with thrombospondin type 1motif,ADAMTS13）为人血管性血友病因子（von Willebrand factor,VWF）裂解酶,主要由肝脏星状细胞合成。近期研究...含I型血小板结合蛋白基序的解聚蛋白样金属蛋白酶（a disintegrin-like and metalloprotease with thrombospondin type 1motif,ADAMTS13）为人血管性血友病因子（von Willebrand factor,VWF）裂解酶,主要由肝脏星状细胞合成。近期研究发现,血管内皮细胞、巨噬细胞、血小板和肾脏多种细胞均表达ADAMTS13。展开更多
The present study examined von Willebrand factor (vWF) levels and ADAMTS13 activity in pregnant and severe preeclamptic women in order to shed light on the prothrombotic state in severe preeclampsia.Thirty healthy wom...The present study examined von Willebrand factor (vWF) levels and ADAMTS13 activity in pregnant and severe preeclamptic women in order to shed light on the prothrombotic state in severe preeclampsia.Thirty healthy women of childbearing age,22 second trimester pregnant women,30 third trimester pregnant women and 10 severe preeclamptic patients were recruited in this study.ADAMTS13 activity was determined by the FRETS-vWF73 assay and vWF antigen (vWF:Ag) levels by an enzyme-linked immunosorbent assay.The results showed that there were statistically significant differences in plasma vWF antigen levels between the severe preeclamptic and third trimester pregnant women,between third and second trimester pregnant women (P【0.05).The third trimester pregnant women had significantly lower plasma ADAMTS13 activity than second trimester pregnant women (P【0.05).Nevertheless,no significant differences in plasma ADAMTS13 activity were found between severe preeclamptic patients and the third trimester pregnant women (P】0.05).In conclusion,plasma ADAMTS13 activity is normal in severe preeclampsia despite the increased vWF:Ag levels.Prothrombotic state is involved in the pathogenesis of severe preeclampsia,as a result of endothelial injury.展开更多
BACKGROUND Many advanced hepatocellular carcinoma(HCC) patients are receiving sorafenib treatment. Sorafenib reportedly improves overall survival(OS) significantly in patients with HCC. Prediction of sorafenib respons...BACKGROUND Many advanced hepatocellular carcinoma(HCC) patients are receiving sorafenib treatment. Sorafenib reportedly improves overall survival(OS) significantly in patients with HCC. Prediction of sorafenib response and prognosis in patients with HCC receiving sorafenib treatment are important due to the potentially serious side effects of sorafenib. A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs 13(ADAMTS13) and von Willebrand factor(VWF) are associated with the pathophysiology of liver cirrhosis and HCC through their roles in hypercoagulability; they are also associated with angiogenesis via vascular endothelial growth factor(VEGF). The imbalance between ADAMTS13 and VWF was associated with prognosis of various cancers in patients undergoing chemotherapy.AIM To investigate ADAMTS13 and VWF as potential biomarkers for sorafenib response and prognosis in patients with HCC receiving sorafenib treatment.METHODS Forty-one patients with HCC receiving sorafenib treatment were included in this study. The initial daily sorafenib dose was 400 mg in all patients. ADAMTS13 activity(ADAMTS13:AC), VWF antigen(VWF:Ag), VEGF levels were determined by enzyme-linked immunosorbent assay. Univariate andmultivariate analyses were used to determine predictive factors for sorafenib response and prognosis in patients with HCC receiving sorafenib treatment.RESULTS ADAMTS13:AC was significantly higher in patients with stable disease(SD),partial response(PR), and complete response(CR) than in those with progressive disease(PD)(P < 0.05). In contrast, VWF:Ag and the VWF:Ag/ADAMTS13:AC ratio were significantly lower in patients with SD, PR, and CR than in those with PD(P < 0.05 for both). Multivariate analysis showed that the VWF:Ag/ADAMTS13:AC ratio was the only predictive factor for sorafenib response and ADAMTS13:AC was the only prognostic factor in patients with HCC receiving sorafenib treatment. The patients with a low ADAMTS13:AC(<78.0) had significantly higher VEGF levels than those with a high ADAMTS13:AC(≥ 78.0)(P < 0.05).CONCLUSION The VWF:Ag/ADAMTS13:AC ratio and ADAMTS13:AC are potentially useful biomarkers for sorafenib response and prognosis, respectively, in patients with HCC receiving sorafenib treatment.展开更多
Objective Thrombotic thrombocytopenic purpura(TTP)is a rare and fatal disease caused by a severe deficiency in the metalloprotease ADAMTS13 and is characterized by thrombotic microangiopathy.The present study aimed to...Objective Thrombotic thrombocytopenic purpura(TTP)is a rare and fatal disease caused by a severe deficiency in the metalloprotease ADAMTS13 and is characterized by thrombotic microangiopathy.The present study aimed to investigate the genes and variants associated with TTP in a Chinese population.Methods Target sequencing was performed on 220 genes related to complements,coagulation factors,platelets,fibrinolytic,endothelial,inflammatory,and anticoagulation systems in 207 TTP patients and 574 controls.Subsequently,logistic regression analysis was carried out to identify the TTP-associated genes based on the counts of rare deleterious variants in the region of a certain gene.Moreover,the associations between common variants and TTP were also investigated.Results ADAMTS13 was the only TTP-associated gene(OR=3.77;95%CI:1.82–7.81;P=3.6×10^(-4))containing rare deleterious variants in TTP patients.Among these 8 variants,5 novel rare variants that might contribute to TTP were identified,including rs200594025,rs782492477,c.T1928G(p.I643S),c.3336_3361del(p.Q1114Afs*20),and c.3469_3470del(p.A1158Sfs*17).No common variants associated with TTP were identified under the stringent criteria of correction for multiple testing.Conclusion ADAMTS13 is the primary gene related to TTP.The genetic variants associated with the occurrence of TTP were slightly different between the Chinese and European populations.展开更多
文摘目的分析血清血管性血友病因子特异性裂解酶(a disintegrin-like and metalloprotease with thrombospondin type1 motif,member 13,ADAMTS13)水平与接受体外膜肺氧合(extracorporeal membrane oxygenation,ECMO)治疗患者的预后关系。方法本研究选取2020年1月—2022年6月在湖南医药学院第一附属医院接受ECMO治疗的42例患者,按照ECMO撤离1个月内预后分为生存组(n=29)和死亡组(n=13)。通过分析接受ECMO治疗患者的血清ADAMTS13水平,分析血清ADAMTS13与接受ECMO治疗患者的预后。结果生存组年龄小于死亡组(P<0.001)。2组性别和体表面积比较,差异无统计学意义(P>0.05)。生存组平均动脉压为(54.24±5.81)mmHg、氧合指数为(178.84±29.38)mmHg,均高于死亡组的(52.56±4.42)mmHg、(155.71±23.12)mmHg;生存组急性生理与慢性健康评分(acute physiology and chronic health evaluation-Ⅱ,APACHE-Ⅱ)评分为(22.51±2.62)分,低于生存组的(27.35±3.85)分(P<0.05)。2组左室射血分数和ECMO流量比较,差异无统计学意义(P>0.05)。ECMO治疗后,2组血清ADAMTS13水平升高。生存组ECMO治疗前1 h及治疗后24 h、48 h血清ADAMTS13水平高于死亡组,差异有统计学意义(P<0.001)。ECMO治疗48 h后血清ADAMTS13水平为ECMO辅助治疗患者死亡的独立危险因素(P<0.001)。结论接受ECMO治疗患者的血清ADAMTS13水平降低与预后不佳相关。
文摘目的本研究旨在研究和探讨血管性血友病因子（von Willebrand factor，vwF）和含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶（A disintegrin—like and metalloprotease with thrombospondin type 1 motif，vWF裂解酶，ADAMTS13）的活性在感染性休克和脓毒症患者早期及预后评估方面的临床意义。方法选取2012—06—2013—06收住无锡市人民医院ICU的感染性休克或脓毒症患者40例，根据预后分为存活组20例和死亡组20例，记录其一般情况及入组时APACHE11评分，于发病第1、3、5、7天分别运用ELISA法检测其血浆vWF活性及ADAMTS13活性，各样本均数用方差分析、受试者工作特征（receiver operating characteristic，ROC）曲线分析、参数间进行Spearman相关分析，并绘制ROC曲线探讨vWF活性、ADAMTS13活性与APACHE Ⅱ评分之间的相关性。结果死亡组的vWF活性均明显高于存活组（P〈0．05），ADAMTS13活性明显低于存活组（P〈0．05）；同时死亡组APACHEⅡ评分明显高于存活组（P〈0．05）；存活组在第3天vwF活性升高、ADAMTS13活性下降、APACHEⅡ评分升高，而在第5天和第7天又出现vWF活性持续下降、ADAMTS13活性持续升高、APACHEⅡ评分持续下降，死亡组则呈现vWF活性持续上升、ADAMTS13活性持续下降、APACHEⅡ评分持续升高（P〈0．05）。40例患者的平均vWF活性与APACHEⅡ评分呈正相关（P〈0．01），而ADAMTS13活性与APACHEⅡ评分呈负相关（P〈0．01）：ROC曲线分析，vWF活性和ADAMTS13活性AUC均接近于1，提示能很好地预测患者的预后（P〈0．01）。结论vWF活性、ADAMTS13活性均是判断感染性休克和脓毒症患者预后的可靠指标。在脓毒症的病理过程中，此二者数值的变化与疾病严重程度密切相关，即vWF活性的升高、ADAMTS13活性的降低预示了患者预后差。
文摘微循环障碍参与了多种疾病的进展,例如脓毒症、多器官衰竭等。血管性假性血友病因子（von Willebrand factor,vWF）可使血小板粘附聚集于血管内皮细胞,导致血小板血栓形成;血管性血友病因子特异性裂解酶（a disintegrin-like and metalloprotease with thrombo-spondin type I repeats 13,ADAMTSl3）可裂解vWF多聚体，预防血栓形成.
文摘背景:血管性血友病因子水解蛋白酶13(a disintegrin and metalloproteinase with a thrombospondin type 1motif,member 13,ADAMTS13)可结合并酶切血管性血友病因子,在防止血小板过多聚集和血栓的形成中起到关键作用。重组ADAMTS13通常选择在真核细胞中表达分泌,然而对于其表达纯化目前仍缺乏一个明确高效的途径。目的:寻找合适的宿主细胞,得到稳定表达野生型ADAMTS13和功能增强型ADAMTS13的细胞株,建立高纯度ADAMTS13的纯化方法,进而研究其生物学功能。方法:以野生型ADAMTS13重组质粒为模板,利用位点突变试剂盒得到功能增强型ADAMTS13重组质粒。比较CHO-S细胞、CHO-K1细胞、293T细胞对ADAMTS13的表达效果,挑选出合适的宿主细胞。通过Ni柱亲和层析和分子筛纯化蛋白;采用7.5%SDS-PAGE、Western-blot鉴定野生型ADAMTS13和功能增强型ADAMTS13的纯度及特异性;利用原子力显微镜技术检测野生型及功能增强型ADAMTS13与血管性血友病因子A2的相互作用,鉴定两种蛋白的活性。结果与结论:相对于CHO-S和CHO-K1,293T细胞更适合作为野生型ADAMTS13和功能增强型ADAMTS13的表达宿主;经Ni柱亲和层析和分子筛纯化后,所得到的野生型ADAMTS13和功能增强型ADAMTS13质量浓度分别为103.7,149.7 mg/L,且两种蛋白纯度均约90%;原子力显微镜检测结果显示,野生型ADAMTS13、功能增强型ADAMTS13与血管性血友病因子A2的黏附频率分别为11.37%,14.70%,表明功能增强型ADAMTS13与血管性血友病因子A2的结合亲和力更高,这与近期ADAMTS13构象研究一致。
文摘含I型血小板结合蛋白基序的解聚蛋白样金属蛋白酶（a disintegrin-like and metalloprotease with thrombospondin type 1motif,ADAMTS13）为人血管性血友病因子（von Willebrand factor,VWF）裂解酶,主要由肝脏星状细胞合成。近期研究发现,血管内皮细胞、巨噬细胞、血小板和肾脏多种细胞均表达ADAMTS13。
基金supported by grants from National Natural Sciences Foundation of China(No.30672243,No.30671093)Natural Sciences Foundation of Hubei Province(No.ZR-Y1271)
文摘The present study examined von Willebrand factor (vWF) levels and ADAMTS13 activity in pregnant and severe preeclamptic women in order to shed light on the prothrombotic state in severe preeclampsia.Thirty healthy women of childbearing age,22 second trimester pregnant women,30 third trimester pregnant women and 10 severe preeclamptic patients were recruited in this study.ADAMTS13 activity was determined by the FRETS-vWF73 assay and vWF antigen (vWF:Ag) levels by an enzyme-linked immunosorbent assay.The results showed that there were statistically significant differences in plasma vWF antigen levels between the severe preeclamptic and third trimester pregnant women,between third and second trimester pregnant women (P【0.05).The third trimester pregnant women had significantly lower plasma ADAMTS13 activity than second trimester pregnant women (P【0.05).Nevertheless,no significant differences in plasma ADAMTS13 activity were found between severe preeclamptic patients and the third trimester pregnant women (P】0.05).In conclusion,plasma ADAMTS13 activity is normal in severe preeclampsia despite the increased vWF:Ag levels.Prothrombotic state is involved in the pathogenesis of severe preeclampsia,as a result of endothelial injury.
文摘BACKGROUND Many advanced hepatocellular carcinoma(HCC) patients are receiving sorafenib treatment. Sorafenib reportedly improves overall survival(OS) significantly in patients with HCC. Prediction of sorafenib response and prognosis in patients with HCC receiving sorafenib treatment are important due to the potentially serious side effects of sorafenib. A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs 13(ADAMTS13) and von Willebrand factor(VWF) are associated with the pathophysiology of liver cirrhosis and HCC through their roles in hypercoagulability; they are also associated with angiogenesis via vascular endothelial growth factor(VEGF). The imbalance between ADAMTS13 and VWF was associated with prognosis of various cancers in patients undergoing chemotherapy.AIM To investigate ADAMTS13 and VWF as potential biomarkers for sorafenib response and prognosis in patients with HCC receiving sorafenib treatment.METHODS Forty-one patients with HCC receiving sorafenib treatment were included in this study. The initial daily sorafenib dose was 400 mg in all patients. ADAMTS13 activity(ADAMTS13:AC), VWF antigen(VWF:Ag), VEGF levels were determined by enzyme-linked immunosorbent assay. Univariate andmultivariate analyses were used to determine predictive factors for sorafenib response and prognosis in patients with HCC receiving sorafenib treatment.RESULTS ADAMTS13:AC was significantly higher in patients with stable disease(SD),partial response(PR), and complete response(CR) than in those with progressive disease(PD)(P < 0.05). In contrast, VWF:Ag and the VWF:Ag/ADAMTS13:AC ratio were significantly lower in patients with SD, PR, and CR than in those with PD(P < 0.05 for both). Multivariate analysis showed that the VWF:Ag/ADAMTS13:AC ratio was the only predictive factor for sorafenib response and ADAMTS13:AC was the only prognostic factor in patients with HCC receiving sorafenib treatment. The patients with a low ADAMTS13:AC(<78.0) had significantly higher VEGF levels than those with a high ADAMTS13:AC(≥ 78.0)(P < 0.05).CONCLUSION The VWF:Ag/ADAMTS13:AC ratio and ADAMTS13:AC are potentially useful biomarkers for sorafenib response and prognosis, respectively, in patients with HCC receiving sorafenib treatment.
基金supported by the National Natural Science Foundation of China(No.82003561).
文摘Objective Thrombotic thrombocytopenic purpura(TTP)is a rare and fatal disease caused by a severe deficiency in the metalloprotease ADAMTS13 and is characterized by thrombotic microangiopathy.The present study aimed to investigate the genes and variants associated with TTP in a Chinese population.Methods Target sequencing was performed on 220 genes related to complements,coagulation factors,platelets,fibrinolytic,endothelial,inflammatory,and anticoagulation systems in 207 TTP patients and 574 controls.Subsequently,logistic regression analysis was carried out to identify the TTP-associated genes based on the counts of rare deleterious variants in the region of a certain gene.Moreover,the associations between common variants and TTP were also investigated.Results ADAMTS13 was the only TTP-associated gene(OR=3.77;95%CI:1.82–7.81;P=3.6×10^(-4))containing rare deleterious variants in TTP patients.Among these 8 variants,5 novel rare variants that might contribute to TTP were identified,including rs200594025,rs782492477,c.T1928G(p.I643S),c.3336_3361del(p.Q1114Afs*20),and c.3469_3470del(p.A1158Sfs*17).No common variants associated with TTP were identified under the stringent criteria of correction for multiple testing.Conclusion ADAMTS13 is the primary gene related to TTP.The genetic variants associated with the occurrence of TTP were slightly different between the Chinese and European populations.
