纤维蛋白/原降解产物质控品的研制

纤维蛋白/原降解产物(FDP)测定是诊断弥覆性血管内凝血、原发性纤溶症、抗凝、溶栓药物应用等纤溶系统亢进性疾病诊断、鉴别诊断及疗效观察的重要指标.当纤溶系统亢进时,纤溶酶活性增强,降解血浆中的纤维蛋白原和纤维蛋白,产生分子量大小不等的碎片,包括碎片X、X'、Y、Y'、A、B、C、H、D、D-二聚体等,统称FDP.     

血纤维蛋白原降解产物对急性心肌梗塞预后的影响

为探讨血纤维蛋白原降解产物对急性心肌梗塞预后的影响，对６０例急性心肌梗塞患者进行溶栓治疗期间观察血纤维蛋白原降解产物的浓度，观察纤维蛋白原降解产物与动脉病变支数和治疗结果的关系。结果纤维蛋白原降解产物高、中浓度组冠状动脉多支病变者比低浓度组多，且发生泵衰竭、心源性休克、恶性心律失常及死亡率均高于低浓度组。血纤维蛋白原降解产物浓度与冠脉病变及预后均呈高度正相关。说明血纤维蛋白原降解产物浓度可间接地反映冠状动脉病变的范围及程度，预测心脏事件及临床结果。     

Ro31—8220对纤维蛋白原降解产物诱导的平滑肌细胞增殖的影响

目的：研究纤维蛋白原降解产物的致有丝分裂原活性及一种新型ＰＫＣ抑制剂Ｒｏ３１－８２２０（Ｒｏ）的作用。方法：大鼠主动脉平滑肌细胞增殖采用结晶紫染色法测定。结果：纤维蛋白原降解产物促进大鼠主动脉平滑肌细胞的增殖，Ｒｏ（０．０１￣１μｍｏｌ·Ｌ＾－１）剂量依赖地抑制增殖。结论：Ｒｏ抑制纤维蛋白原降解产物诱导的平滑肌细胞的增殖。     

血浆D-二聚体和纤维蛋白原降解产物在急性脑梗死患者中的检测意义

目的分析血浆D-二聚体（DD）和纤维蛋白原降解产物（FDP）水平检测在急性脑梗死患者中的意义。方法临床确诊的急性脑梗死患者93例（急性脑梗死组）于入院24h、3d、7d抽取空腹静脉血,分别采用ELISA法和Clause法测定DD和FDP水平。选择同期45例健康体检者为对照组。结果急性脑梗死组24h、3dDD水平显著高于对照组（P〈0．05或〈0．01）,7d时较对照组比较差异无统计学意义（P〉0．05）。急性脑梗死组24hFDP与对照组比较差异无统计学意义（P〉0．05）,3d及7d时FDP水平显著高于对照组（P〈0．05或〈0．01）。中、重型组DD、FDP水平分别显著高于轻型组（P〈0．05或〈0．01）,而重型组DD、FDP水平显著高于中型组（P〈0．05）。中梗死灶、大梗死灶DD、FDP水平分别显著高于小梗死灶（P〈0．05或〈0．01）,而大梗死灶DD、FDP水平显著高于中梗死灶（P〈0．05）。急性脑梗死患者DD水平与FDP水平呈显著正相关（r=0．71,P〈0．05）。结论急性脑梗死患者血浆DD和FDP水平呈不同程度的增高,动态监测患者血浆DD和FDP水平对于判断脑梗死的严重程度和梗死病灶大小具有重要临床意义。     

纤维蛋白（原）及其降解产物与缺血性心脏病

血栓形成学说在缺血性心脏病（IHD）的发病机制中起着重要的作用。如何预防高凝状态的产生对防治IHD有着重要的意义。许多学者通过前瞻性研究，IHD的基础和临床的研究，指出纤维蛋白原（Fg）、纤维蛋白肽A（FPA）和交联的纤维蛋白降解产物D－H聚体（DD）在凝血及纤溶系统产生变化时而发生量的改变，通过监测它们的变化，对IHD的发病机制的研究，病变程度的预测、治疗、疗效的观察及预后判断均具有重要意义。一、生物学特性血浆rg是由肝脏合成的一种糖蛋白，由三个不同的亚单位（二条Aa、二条BS和二条Y链）组成，分子量为34000O。…     

不同类型冠心病患者血浆D-二聚体、纤维蛋白原含量的变化

目的 :探讨D -二聚体、纤维蛋白原 (Fg)水平以评估不同类型冠心病患者凝血、纤溶活性的临床意义。方法 :采用定量酶联免疫吸附法和全自动血凝分析仪检测冠心病患者126例 ,其中急性心肌梗死 (AMI)42例 ,不稳定型心绞痛 (UA)36例 ,稳定型心绞痛 (SA)48例和30例正常对照组的D -二聚体及纤维蛋白原水平。结果 :D -二聚体和Fg的阳性发生率及含量,UA和AMI患者组明显高于SA组及对照组(P<0.01) ;SA组患者的D -二聚体和Fg与对照组比较差别无统计学意义 (P>0.05) ;UA组D -二聚体阴性组的Fg水平小于阳性组 (P<0.05) ,而AMI这两组Fg水平差别无统计学意义 (P>0.05)。结论 :冠心病患者D -二聚体及Fg 含量越高 ,越易发生AMI ,UA组D -二聚体、Fg增高者可有血栓形成。D -二聚体和Fg含量测定可作为冠心病患者发生新鲜血栓形成的指标     

纤维蛋白原降解产物和血浆D-二聚体在急性脑梗死患者中的检测意义

目的探讨血浆D-二聚体和纤维蛋白原降解产物在急性脑梗死患者中的检测意义。方法选择本院2009年10月～2011年10月急性脑梗死患者30例,所选患者均度过急性期（发病72h内）和恢复期（治疗2周后）。同时选取本院健康体检者共30例为对照组。对所选脑梗死患者和健康体检者抽取静脉血检测D-二聚体和纤维蛋白原降解产物（FDP）水平。结果急性脑梗死的急性期FDP和D-二聚体水平均高于对照组,差异有统计学意义（P〈0.05）;急性脑梗死恢复期FDP水平与对照组比较,差异无统计学意义（P〉0.05）;急性脑梗死的急性期FDP、D-二聚体水平均高于急性脑梗死的恢复期,差异有统计学意义（P〈0.05）。结论血浆D-二聚体和FDP联合检测有助于判断急性脑梗死的发生、脑梗死的严重程度及观察疗效,具有较高的检测价值,值得借鉴。     

血浆血管性血友病因子抗原、D-二聚体、纤维蛋白原/纤维蛋白降解产物检测在急性脑梗死中的诊断价值

目的 探讨血浆血管性血友病因子抗原(vWF∶Ag)、D-二聚体(D-D)、纤维蛋白原/纤维蛋白降解产物(FDP)检测在急性脑梗死中的诊断价值.方法 选取2019年4月至2020年12月上海仁济医院宝山分院收治的86例急性脑梗死患者作为观察组,选取2019年4月至2020年12月上海仁济医院宝山分院60例健康体检者为对照...     

不同病理类型狼疮性肾炎血尿纤维蛋白降解产物水平分析

目的 探讨不同病理类型狼疮性肾炎(lupus nephritis,LN)患者血、尿纤维蛋白降解产物(fibrinogen degradation products,FDP)水平及临床意义.方法 用酶联免疫吸附试验(ELISA法)测定140例不同病理类型LN血、尿FDP水平并与健康体检者(正常对照组)进行比较.结果 LN组血、尿FDP水平高于正常对照组(P＜0.01).LN组不同病理类型血、尿FDP水平均高于正常对照组,差异有统计学意义(P<0.01);Ⅱ型血FDP水平低于Ⅳ型,Ⅴ、Ⅳ+Ⅴ型血、尿FDP水平均低于Ⅳ型,差异均有统计学意义(P<0.05).结论 不同病理类型LN患者血、尿FDP水平存在差异,对了解不同病理类型LN患者凝血-纤溶情况及疾病严重程度具有重要意义,同时也为诊断及治疗提供了参考.     

Snake venom fibrin(ogen)olytic enzymes

Snake venoms contain a number of serine and metalloproteinases and included among these are the fibrin(ogen)olytic proteinases. Some years ago it was postulated that the fibrin(ogen)olytic enzymes may be clinically useful. Over the past 150 years a substantial body of literature has been generated on the identification and characterization of fibrin(ogen)olytic enzymes from a broad spectrum of snake species. In this review we describe the two different classes of fibrin(ogen)olytic enzymes isolated from snake venom and we summarize a number of studies aimed at characterizing the purified enzymes and/or their derivatives.

Two distinct classes of venom fibrin(ogen)olytic enzymes have been previously identified, the metalloproteinases and serine proteinases. These two classes of proteinases differ in their mechanism of action and they target different amino acid sequences in fibrin(ogen), but each perform the same role in nature. When a snake envenomates its prey it needs a mechanism to facilitate the spread of the toxic components throughout the circulation. Fibrin(ogen)olytic enzymes break down fibrin rich clots and help to prevent further clot formation by their action on fibrinogen. This characteristic feature has led to development of fibrin(ogen)olytic snake venom enzymes as potential clinical agents to treat occlusive thrombi. Fibrolase, a fibrinolytic metalloproteinase isolated from Agkistrodon contortrix contortrix venom and the serine β-fibrinogenolytic proteinase from Vipera lebetina have been chosen as representative enzymes from the two classes, and their biochemical and physiochemical properties will be described in detail.

Finally, the characterization and development of alfimeprase, a recombinant fibrinolytic enzyme derived from fibrolase, as a clinical agent is described citing the progression from the laboratory bench to its current status as having successfully completed Phase II clinical trials.     

Procoagulant activity of Calotropis gigantea latex associated with fibrin(ogen)olytic activity.

The latex of Calotropis gigantea is a rich source of useful components that has medicinal properties and one of the main applications is in controlling bleeding. The crude latex extract contained many proteins, which are highly basic in nature and exhibited strong proteolytic activity. The crude extract hydrolyses casein, human fibrinogen and crude fibrin clot in a dose-dependent manner. The hydrolyzing activity was completely inhibited by IAA indicating they belong to the super family, cysteine proteases. Crude extract hydrolyses Aalpha, Bbeta and gamma subunits of fibrinogen. Among all the subunits the preferential subunit to get hydrolyzed was Aalpha followed by Bbeta and gamma subunit is highly resistant and hydrolyzed at higher protein concentration or over a prolonged incubation time. The crude extract hydrolysis crude fibrin clot strongly compared to trypsin and papain. Pharmacologically the crude extract is hemorrhagic and induces skin hemorrhage at >75 microg and reduces the coagulation time of citrated plasma from 150 to 47 s and promotes blood coagulation. Procoagulation and blood clot hydrolysis are important in wound healing process. This is due to unique cysteine proteases of plant latex and is responsible for the pharmacological actions observed in folk medicine.     

cDNA cloning, expression and fibrin(ogen)olytic activity of two low-molecular weight snake venom metalloproteinases

Two cDNA clones, AplVMP1 and AplVMP2, were isolated from a snake (Agkistrodon piscivorus leucostoma) venom gland cDNA library. The full-length cDNA sequence of AplVMP1 with a calculated molecular mass of 46.61 kDa is 1233 bp in length. AplVMP1 encodes PI class metalloproteinase with an open reading frame of 411 amino acid residues that includes signal peptide, pro-domain and metalloproteinase domains. The full-length cDNA of the AplVMP2 (1371 bp) has a calculated molecular mass of 51.16 kDa and encodes PII class metalloproteinase. The open reading frame of AplVMP2 with a 457 amino acid residues is composed of signal peptide, pro-domain, metalloproteinase and disintegrin domains. AplVMP1 and AplVMP2 showed 85% and 93% amino acid identical to PI class enzyme Agkistrodon contortrix laticinctus ACLPREF and PII class enzyme Agkistrodon piscivorus piscivorus piscivostatin, respectively. When expressed in Escherichia coli, most of recombinant proteins of AplVMP1 and AplVMP2 were in insoluble inclusion bodies, with soluble yields of 0.7 mg/l and 0.4 mg/l bacterial culture, respectively. Both affinity purified recombinant proteins show proteolytic activity on fibrinogen, although having an activity lower than that of crude A. p. leucostoma venom. Proteolytic activities of AplVMP1 and AplVMP2 were completely abolished after incubation with a final concentration of 100 μM of EDTA or 1,10-phenanthroline. Both AplVMP1 and AplVMP2 were active in a fibrin-agarose plate but devoid of hemorrhagic activity when injected (up to 50 μg) subcutaneously into mice, and had no capacity to inhibit platelet aggregation.     

卡维地洛在运动诱发和动态心电图监测时对心肌缺血的影响

目的　观察卡维地洛与非洛地平治疗在运动诱发和动态心电图监测时对心肌缺血的影响。方法　对 110例活动平板运动试验与动态心电图监测有心肌缺血的病人分为两组 ,分别服用卡维地洛和非洛地平 4周 ,观察治疗后效果。结果　两组均可提高抗心肌缺血效应和增加病人运动耐量 ,但卡维地洛提高病人运动耐量时间明显优于非洛地平 (P <0 0 5 )。结论　卡维地洛可提高抗心肌缺血效应 ,安全性好。     

Origin of urinary fibrin/fibrinogen degradation products in glomerulonephritis.

To elucidate the origin of the fibrin/fibrinogen degradation products (F.D.P.) occurring in the urine in glomerulonephritis 28 patients with glomerulonephritis were examined for renal fibrinolytic activity, F.D.P. in urine and serum, and blood fibrinolytic activators and blood fibrinolytic activators and inhibitors. Unlike the glomerful of healthy kidneys, which were fibrinolyticly inactive, those of kidneys with glomerulonephritis constantly showed fibrinolytic activity. The presence or absence of fibrin in the glomeruli was almost always accompanied by, respectively, the presence or absence of urinary F.D.P., which suggested a renal origin of urinary F.D.P. in glomerulonephritis. The low fibrinolytic activity of the blood and the absence of F.D.P. in the serum of these patients make it unlikely that the urinary F.D.P. in glomerulonephritis result from glomerular filtration.     

Isolation and structural characterization of a new fibrin(ogen)olytic metalloproteinase from Bothrops moojeni snake venom.

A proteinase, named BmooMPalpha-I, from the venom of Bothrops moojeni, was purified by DEAE-Sephacel, Sephadex G-75 and heparin-agarose column chromatography. The enzyme was purified to homogeneity as judged by its migration profile in SDS-PAGE stained with coomassie blue, and showed a molecular mass of about 24.5 kDa. Its complete cDNA was obtained by RT-PCR and the 615 bp codified for a mature protein of 205 amino acid residues. The multiple alignment of its deduced amino acid sequence and those of other snake venom metalloproteinases showed a high structural similarly, mainly among class P-IB proteases. The enzyme cleaves the Aalpha-chain of fibrinogen first, followed by the Bbeta-chain, and shows no effects on the gamma-chain. On fibrin, the enzyme hydrolyzed only the beta-chain, leaving the gamma-dimer apparently untouched. It was devoid of phospholipase A(2), hemorrhagic and thrombin-like activities. Like many venom enzymes, it is stable at pH values between 4 and 10 and stable at 70 degrees C for 15 min. The inhibitory effects of EDTA on the fibrinogenolytic activity suggest that BmooMPalpha-I is a metalloproteinase and inhibition by beta-mercaptoethanol revealed the important role of the disulfide bonds in the stabilization of the native structure. Aprotinin and benzamidine, specific serine proteinase inhibitors, had no effect on BmooMPalpha-I activity. Since the BmooMPalpha-I enzyme was found to cause defibrinogenation when administered i.p. on mice, it is expected that it may be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis.