高半胱氨酸对血管内皮细胞,血小板聚集和肝素辅助因子活性？…

目的 揭示高半胱氨酸（Ｈｃｙ）致血栓形成的机制。方法 （１）免疫荧光分析务管内皮细胞凝血酶调节蛋白（ＴＭ）抗原分布;（２）发色基质法测定肝素辅助因子Ⅱ（ＨＣⅡ）活性。结果 （１）Ｈｃｙ可下调血管内皮细胞表面ＴＭ的表达,且呈浓度依赖性;（２）Ｈｃｙ对血浆ＡＴ－Ⅲ和ＨＣⅡ的抗凝血酶活性无明显影响,在纯化的ＡＴ－Ⅲ的抗凝血酶作用也无影响。结论 Ｈｃｙ的致血栓效应与其对内皮细胞ＴＭ表达调节有关,且独立于血     

α2巨球蛋白在儿童期的抗凝血作用

α＿２巨球蛋白在儿童期的抗凝血作用天津儿童医院儿科研究所（３０００７４）侯秀荣（综述）中国医学科学院血液学研究所包承鑫（审校）人体重要的抗凝血酶包括：抗凝血酶Ⅲ（ＡＴ－Ⅲ）、α＿２巨球蛋白（α＿２Ｍ）、肝素辅因子Ⅱ（ＨＣⅡ）。在成人ＡＴ－Ⅲ起主要抗凝...     

高半胱氨酸对血管内皮细胞、血小板聚集和肝素辅助因子活性效应的实验研究

目的　揭示高半胱氨酸（ Ｈｃｙ） 致血栓形成的机制。方法　①免疫荧光分析血管内皮细胞凝血酶调节蛋白（ Ｔ Ｍ） 抗原分布；②发色基质法测定肝素辅助因子Ⅱ（ Ｈ ＣⅡ） 活性。结果　① Ｈｃｙ 可下调血管内皮细胞表面 Ｔ Ｍ 的表达，且呈浓度依赖性；② Ｈｃｙ 对 Ａ Ｄ Ｐ 和凝血酶诱导的血小板聚集均无影响；③在正常人血浆体系中， 以肝素和硫酸皮肤素（ Ｄ Ｓ） 为激动剂， Ｈｃｙ 对血浆 Ａ ＴⅢ和 Ｈ ＣⅡ的抗凝血酶活性无明显影响， 在纯化的 Ａ ＴⅢ体系， Ｈｃｙ 对 Ａ ＴⅢ的抗凝血酶作用也无影响。结论　 Ｈｃｙ 的致血栓效应与其对内皮细胞 Ｔ Ｍ 表达调节有关，且独立于血小板聚集功能和肝素辅助因子活性变化。     

紫外速率直接法测定过氧化氢酶活性

紫外速率直接法（１）测定过氧化氢酶（ｃａｔａｌａｓｅ，ＣＡＴ）是一种简便、快速、的测定方法。本法在２４０ｎｍ测定Ｈ２Ｏ２被ＣＡＴ催化分解时，吸不度的降低。由于ＣＡＴ催化Ｈ２Ｏ２的分解反应在一定底物浓度范围符合一级反应，因此计算出速常数Ｋ，即代表ＣＡＴ活性。本法批内变异２．３４％，日间变异７．７８％，测定范围５．７－４３８．７Ｋ／Ｌ，平均回收率１００．５３％，正常参考值范围２８１．３２－５３３．６１     

白细胞介素4与血管活性多肽对大鼠血管平滑肌细胞增殖的调节作用

以＾３Ｈ－ＴｄＲ掺入法观察内皮素（ＥＴ）、血管紧张素Ⅱ及白细胞介素４（ＩＬ－４）单独作用和联合作用时对大鼠血管平滑肌细胞（ＶＳＭＣ）增殖的影响。结果发现ＩＬ－４能抑制培养的自发性高血压大鼠（ＳＨＲ）血管平滑肌细胞（ＶＳＭＣ）的＾３Ｈ－ＴｄＲ的掺入（最低浓度为５ｎｇ／ｍｌ），并存在剂量效应关系，而对正常血压大鼠（ＷＫＹ）的ＶＳＭＣ无影响；但预先以ＥＴ或ＡＧ－Ⅱ刺激ＷＫＹ的ＶＳＭＣ的增殖，则ＩＬ－４表     

紫热线照射充氧自血中输对冠心病患者血清脂蛋白及载体蛋白的 …

目的：探讨紫外线照射充氧自血回输（ＵＢＩＯ）对血清总胆固醇（Ｔｃｈ）、甘油三酯（ＴＧ）、高密度脂蛋白胆固醇（ＨＤＬ－Ｃ）及其亚组分Ⅱ（ＨＤＬ２－Ｃ）、亚组分Ⅲ（ＨＤＬ３－Ｃ）、低密度脂蛋白胆固醇（ＬＤＬ－Ｃ）、载体蛋白（Ａｐｏ）Ａ１、Ａ２、ｂ１００、Ｃ２、Ｃ３的影响．方法：将冠心病高血脂患者分成两组，对照且２７例，给予硝酸酯类和维生素类药物，ＵＢＩＯ组３２中用光量子疗法。两组患者治疗期间停用其他影     

维甲酸诱导HL-60细胞分化中对细胞周期素依赖性激酶活性的影响

目的探讨维甲酸（ＲＡ）对ＨＬ６０细胞的增殖抑制和诱导分化作用与细胞周期素依赖性激酶（ＣＤＫ）活性变化的相关性。方法用５×１０－６ｍｏｌ／Ｌ全反式维甲酸（ＡＴＲＡ）或芳维甲（ＡＥ）处理ＨＬ６０细胞１～４天,在观察细胞生长、四氮唑蓝（ＮＢＴ）还原实验以及细胞周期分析的基础上,用组蛋白Ｈ１激酶活性分析技术检测ＣＤＫ２的活性,免疫沉淀法分析结合于ＣＤＫ２和ＣＤＫ４上的相应的细胞周期素（ｃｙｃｌｉｎ）Ｅ和Ｄ的量。结果ＨＬ６０细胞在ＡＴＲＡ或ＡＥ作用下,细胞增殖减慢,对ＮＢＴ的还原能力明显增强,９０％左右的细胞被阻滞在Ｇ１／Ｇ０期;在Ｇ１到Ｓ期转换过程中起关键作用的ｃｙｃｌｉｎＥ／ＣＤＫ２的活性显著下降,结合于ＣＤＫ４上的ｃｙｃｌｉｎＤ１的量减少。结论ＲＡ抑制ＨＬ６０细胞增殖并诱导其分化可能与ＣＤＫ２和ＣＤＫ４的活性下降有密切关系     

江苏徐州地区丙型肝炎病毒基因型研究

为了解本地区丙型肝炎病毒（ＨＣＶ）基因型分布情况，用限制性片段长度多态性分析法（ＲＦＬＰ），对１０７便丙型肝炎患者中血清ＨＣＶ－ＲＮＡ阳性的９０例进行基因型分析。分型结果表明，ＨＣＶ－Ⅱ（１ｂ）占７８．９％，ＨＣＶ－Ⅲ（２ａ）型占１８．９％，ＨＣＶ－Ⅱ／Ⅲ混合型占２．２％，提示：（１）本地区ＨＣＶ流行株基因主要为Ⅱ型，同时存在Ⅲ型与Ⅱ／Ⅲ混合型。徐州地区ＨＣＶ基因型分布符合我国ＨＣＶ基因型分布的特     

Graves‘病人TSH抗独特型抗体和TRAb相关性研究

根据自身免疫性甲状腺疾病（Ａｕｔｏｍｉｍｍｕｎｏ ｔｈｙｒｏｉｄ ｄｉｓｅａｓｅ，ＡＩＴＤ）发病机制新的独特型－抗独特型免疫网络学说，用ＴＳＨ独特型抗体（ＴＳＨＡｂ１）检测ＴＳＨ抗独特抗体（ＴＳＨＡｂ２），用＾１２５Ｉ－ｈＴ－ＳＨＡｂ１＾１２５Ｉ－１３Ｂ４（ｈＴＳＨ单抗）检测ＴＲＡｂ阳性和阴性的Ｇｒａｖｅｓ‘病人，依正常混合血清为质控标本，依ＴＡｂ阴性组结合率ｘ＋２ｓ为标准，大于此值为ＴＳＨＡｂ２     

肝素激活抗凝血酶Ⅲ的机制研究进展

肝素激活抗凝血酶Ⅲ的机制研究进展６１００８１中国医学科学院中国协和医科大学输血研究所刘晓宇抗凝血酶Ⅲ（ＡＴ Ⅲ）是体内重要的抗凝血因子，属于单链血浆糖蛋白，分子量５８２００ＫＤ，其主要功能是抑制活化的Ⅱ因子（凝血酶），也可以抑制其它丝氨酸蛋白酶如Ⅶａ...     

Structure-activity relationships of heparin. Independence of heparin charge density and antithrombin-binding domains in thrombin inhibition by antithrombin and heparin cofactor II.

To better understand how heparin structure affects its activity the relationships between the functional domains for inhibitor binding and charge density were investigated to determine how these domains affect heparin-mediated thrombin inhibition by two different heparin-dependent protease inhibitors, antithrombin (AT) and heparin cofactor II (HC II). A series of heparins, fractionated systematically by charge density, was further fractionated on antithrombin agarose to isolate more homogeneous subfractions that were either inactive or highly active with respect to thrombin inhibition by AT. With AT, the activities of the AT-active subfractions increased sharply with heparin charge density, while those with little or no affinity for AT were virtually inactive. In contrast, with HC II inhibitor, the activities of the heparins depended only upon their charge densities and were independent of AT affinity. At any given charge density, the heparin before fractionation by AT affinity and the fractions that were highly active and inactive with AT were all equally active with HC II. The two inhibitors also differed in their reactivity with heparan sulfate and dermatan sulfate. A charge-density effect with the subfractions having similar high affinity for AT demonstrates that charge density represents a heparin functional domain that is independent of the AT-binding domain. The behavior of the AT-inactive heparins, being fully active with HC II, demonstrates the functional domain necessary for AT binding is not needed to produce HC II activity.     

A kinetic micromethod of determining the activity of antithrombin III

A method for measuring the blood plasma antithrombin III (AT III) activity is suggested, based on measurement of the rate of inactivation of added standard volume (0.05 ml) of plasma thrombin AT III. The thrombin residual activity in the mixture is detectable over the course of fibrinogen tests. Curves are plotted, reflecting the rate of inactivation of thrombin AT III in the tested and reference plasma samples; by modeling and comparing these curves the values of the examined sample AT III activity are obtained, with the effects of fibrinogen and fast antithrombins various levels eliminated.     

Involvement of heparin cofactor II in chymotrypsin neutralization and in the pancreatic proteinase—antiproteinase interaction during acute pancreatitis in man

Heparin cofactor II is a proteinase inhibitor which inhibits both chymotrypsin and thrombin, and displays great similarities with antithrombin III, the main inhibitor of thrombin in human plasma. Since acute pancreatitis is known to be associated with modification of the proteinase-antiproteinase equilibrium, we studied heparin cofactor II and antithrombin III as well as other biochemical and haematological parameters in 10 patients experiencing attacks of acute pancreatitis. Heparin cofactor II activity decreased during the first week of illness, while its antigen concentration remained subnormal. This discrepancy between antigen concentration and activity which persisted during the first week of illness was due both to complex formation of heparin cofactor II with its target proteinases and to partial proteolysis of the inhibitor. Heparin cofactor II was shown to form a complex with chymotrypsin in the plasma of such patients. Antithrombin III levels remained unchanged throughout the study, with no discrepancy between its activity and antigen concentration. No modification of haemostasis was shown either, except for a rise in the fibrinogen level during the first days of illness. It is concluded that, unlike antithrombin III, heparin cofactor II is involved in the proteinase-inhibitor equilibrium in patients with acute pancreatitis, and that heparin cofactor II might react as an inhibitor of pancreatic proteinases rather than an inhibitor of thrombin.     

Effect of protamine sulfate on antithrombin III activity

When protamine sulfate was added to heparinized plasma in vitro for neutralization of heparin, the activities on both thrombin and Xa known as heparin cofactor in antithrombin action were completely abolished. However, progressive activities on thrombin and Xa both recovered within 30 minutes after protamine sulfate addition. When equivalent heparin was again added, heparin cofactor activity was immediately restored. Based on the fact that protamine sulfate did not show any direct action on the antithrombin III molecule, the presence of AT III with progressive activity was considered to play an important role in the rebound phenomenon of heparin after heparin neutralization with protamine sulfate.     

脑梗死患者血浆中血栓标志物及凝血指标联合检测的临床意义

目的探讨凝血分子标志物和常规凝血指标的检测在脑梗死患者中的临床意义,为脑梗死的诊断和治疗监测提供依据。方法测定90例脑梗死患者血浆中凝血酶原片段1 2(F1 2)、凝血酶-抗凝血酶复合物(TAT)、二聚体(D-D)、抗凝血酶(AT)、蛋白C(PC)、血管性血友病因子(vWF)、凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、凝血酶时间(TT),并与健康正常对照组进行比较。结果脑梗死患者F1 2、TAT、D-D、vWF和健康对照组相比均有显著增高,而PT、APTT、TT、PC和AT与健康对照组相比差异无统计学意义。结论脑梗死患者体内呈高凝状态,凝血酶原活性增强,凝血酶生成增多,纤溶活性增强,抗凝系统活化不足,同时内皮细胞损伤在脑梗死患者的凝血系统激活和发病过程中可能起主要作用,F1 2、TAT、D-D、vWF等凝血分子标志物可以作为脑梗死的诊断指标,而常规的凝血指标不能反映脑梗死患者的高凝状态。     

The influence of antithrombin III (AT III) substitution to supranormal activities on systemic procoagulant turnover in patients with end-stage chronic liver disease

Objective: Since antithrombin III (AT III) substitution to normal activities could not be shown to have major beneficial effects in patients with end-stage chronic liver disease in a variety of clinical settings, we tested the hypothesis that substitution to supranormal activities decreases systemic procoagulant turnover better in this patient group. Design: Controlled prospective clinical study. Setting: Operating rooms at a University Hospital. Patients: Twenty-four patients with histologically verified liver cirrhosis consecutively scheduled for liver transplantation. Interventions: Nineteen patients were given an antithrombin III concentrate to achieve either 100 % (n = 10) or 175 % (n = 9) AT III activity. Control patients (n = 5) received saline 0.9 % instead. Measurements and results: Molecular markers of coagulation activation, platelet count and aggregability, and global coagulation variables were measured prior to AT III infusion and 60 min thereafter. In both AT III-treated groups thrombin-antithrombin III-complex increased significantly (p < 0.005), whereas prothrombin fragment F1 + 2, soluble fibrin and D-dimer concentrations, as well as other variables, did not show major changes. Conclusions: Despite thrombin inhibition by AT III in patients with end-stage chronic liver disease, systemic procoagulant turnover was not significantly decreased 60 min after AT III application even to supranormal activities. Replenishment of the inhibitory antithrombin III pool, decreased in chronic liver disease, should not be expected to slow down the baseline consumptive component of the haemostatic disorder in this patient group. Received: 23 December 1996 Accepted: 4 September 1997     

Internalization of antithrombin III by cultured human endothelial cells and its subcellular localization

The presence of antithrombin III was demonstrated in cultured human endothelial cells derived from the umbilical cord by using immunofluorescence, immunoelectron microscopy studies, and an enzyme-linked immunosorbent assay (ELISA) specific for antithrombin III. Immunofluorescence studies indicated the presence of antithrombin III in granule-like structures in the endothelial cell. Immunoelectron microscopy studies performed with ultrathin cryosections of endothelial cells showed a colocalization of antithrombin III and a lysosomal marker protein in low electron dense organelles, indicating a lysosomal localization of antithrombin III. By using the ELISA, 77 +/- 40 ng (n = 8) antithrombin III was quantitated in 10(6) endothelial cells. Immunoprecipitation studies performed with metabolically labeled cultured human endothelial cells indicated that antithrombin III was not synthesized by the cells. Endothelial cells cultured in antithrombin III-depleted human serum did not contain antithrombin III, as was measured by ELISA. Internalization studies performed with radiolabeled purified antithrombin III and antithrombin III-thrombin complexes indicated that endothelial cells internalize antithrombin III when it is complexed to thrombin. Antithrombin III alone was not internalized by the endothelial cells.     

Metabolic control may alter antithrombin III activity but not its plasma concentration in diabetes: a possible role for nonenzymatic glycosylation

The effects of metabolic control on both antithrombin III (AT III) activity and AT III plasma concentration in 20 insulin-treated diabetic subjects have been evaluated. Basal AT III activity was significantly lower in diabetic subjects versus healthy controls (P less than 0.001), whereas no difference was found in AT III concentration. A good correlation was found between AT III activity and AT III concentration (r = 0.81; P less than 0.001) in healthy controls, but this correlation was not significant in diabetic subjects (r = 0.12; P = NS). In those subjects a linear inverse correlation was found to exist between AT III activity and level of glycosylated proteins (r = -0.43; P less than 0.05). Diabetic subjects were also examined after 1 and 2 mo of restored metabolic control, obtained by human insulin (DNA-recombinant) therapy. Improved metabolic control was characterized by an increase of AT III activity (P less than 0.05), a decrease of mean daily blood glucose, and stable HbA1 and glycosylated proteins (P less than 0.05), while AT III concentration did not vary. On the other hand, a significant inverse correlation between AT III activity and glycosylated proteins was found during both the first and second months (r = -0.54 and r = -0.53, respectively; P less than 0.01). Moreover, no correlation between AT III activity and AT III concentration was found. These data suggest that impaired metabolic control may alter the biologic activity of AT III in diabetes, but not its plasma concentration.     

A sensitive colorimetric assay for thrombin, prothrombin and antithrombin III in human plasma using a new synthetic substrate

Benzoyl-L-leucyl-L-alanyl-L-arginine-alpha-naphthylester (Bz-Leu-Ala-Arg-NE) was synthesized as a new substrate for use in the assay of thrombin. In the assay alpha-naphthol released by the enzyme reaction was measured colorimetrically. With Bz-Leu-Ala-Arg-NE as substrate, the minimum detectable concentration of human thrombin was 0.0025 U. This assay using Bz-Leu-Ala-Arg-NE is a highly sensitive method for detecting prothrombin, thrombin and antithrombin III in human plasma. Prothrombin could be determined with 0.2 microliter of human plasma using Echis carinatus venom (ECV) as activator. Antithrombin III activity could be determined with 2 microliter of human plasma using human thrombin and heparin as cofactor. A zymogram of human prothrombin was prepared with Bz-Leu-Ala-Arg-NE as substrate. The preparation gave one band (pI 4.9) on polyacrylamide disc gel isoelectrophoresis.