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介绍一种改良的用以快速地定性和半定量检测纤溶活性蛋白的纤维平板法.对该方法中的几个关键因素作了详细的研究和探讨.纤维平板中纤维蛋白原和纤溶酶原的浓度对裂解面积不产生任何影响,但是纤维蛋白原的浓度会对裂解圈的观察产生影响,纤维蛋白原浓度低于02 mg/mL时会使背景色过淡而不能清晰地观察到裂解圈.尿激酶浓度在2~250 mU/μL范围时,尿激酶浓度的对数和裂解圈相互垂直的两条直径乘积的对数之间存在明显的线性关系.在本实验设计方案下,尿激酶的最佳使用浓度为10~20 mU/μL.同时,本文还探讨了纤维平板中纤溶酶原的去活条件,当琼脂纤维平板凝固后很难完全去活纤溶酶原,但是,在琼脂纤维蛋白原混合物凝固前于95 ℃持续温浴40 min,则能彻底去活纤维蛋白原中残留的纤溶酶原. 相似文献
3.
Shiang-Cheng Kung Bonapally Aravind Stephen Morse Larry E. Jacobs Rasib Raja 《Hemodialysis international. International Symposium on Home Hemodialysis》2001,5(1):32-36
Tunneled dialysis catheter–associated right atrial thrombus (RAT) is a rarely reported complication. We reviewed hospital records of 10 patients from a teaching hospital dialysis unit, in whom RAT was diagnosed by trans‐esophageal echocardiography (TEE). Patients were treated with chronic anticoagulation (heparin followed by warfarin) and followed over time. The group included 7 women; 6 patients were African American, 3 were Caucasian, and 1 was Hispanic. The average age was 52.1 ± 15.3 years. The most common presenting symptom was poor catheter flow on hemodialysis followed by fever and chills. On average, the patients had had 3.4 ± 2.7 catheter insertions before diagnosis of RAT, and the tunneled dialysis catheter (TC) had been in place for a mean of 91 ± 89.4 days when the thrombi were diagnosed. Trans‐thoracic echocardiography (2‐D echo) was done in 4 patients, but it identified RAT in only 1 patient. The catheter tip was at the junction of the superior vena cava and right atrium (SVC/RA) in most patients. Thrombolysis (unsuccessful) was attempted with urokinase in 3 patients, complicated in 2 patients by hemorrhage. After anticoagulation, 90% of the RAT resolved on repeated TEE. One patient had persistent RAT for 23 weeks and underwent surgical thrombolysis, but died postoperatively. We conclude that RAT is a frequently missed complication of a TC. Positioning the tip of the TC at the SVC/RA junction may not prevent RAT. Trans‐esophageal echocardiography is a more sensitive diagnostic tool than 2‐D echo and should be obtained early. Most patients can be successfully treated with anticoagulation alone. Thrombolytic therapy and surgical thrombolysis have high morbidity and mortality. 相似文献
4.
Bakker Arjen H. F.; Nieuwenbroek N. M. E.; Verheijen J. H. 《Protein engineering, design & selection : PEDS》1995,8(12):1295-1302
Fibrin-dependent plasminogen activation by tissue-type plasminogenactivator (t-PA) is in part associated with the presence ofthe kringle 2 domain in t-PA. Within this kringle 2 domain alysyl-binding site has been described. The plasminogen to plasminconversion by urokinase-type plasminogen activator (u-PA), incontrast to that of t-PA, is not enhanced in the presence offibrin. Within the u-PA kringle domain no lysyl-binding siteis found. To study whether introduction of a lysyl-binding sitein the u-PA kringle domain will make u-PA a fibrin-dependentplasminogen activator, three stretches of amino acid residuesof the u-PA kringle domain (A28-Q33, D55-N57 and G67-V72) weresubstituted by three stretches of amino acids from the correspondingpositions of the kringle 2 domain of t-PA (M28-K33, D55-D57and N67-W72). These changes resulted in the creation of thelysyl-binding site consensus of the kringle 2 domain (K33, D55,D57, W62 and W72) in the u-PA kringle. However, the resultingu-PA mutant did not interact with lysyl-Sepharose, nor did itdisplay fibrin-enhanced plasminogen activation in the presenceof soluble fibrin mimic. When the kringle domain of u-PA wasreplaced by the kringle 2 domain of t-PA, similar results wereobtained. The hybrid protein hardly interacted with lysyl-Sepharoseand the plasminogen activation was not enhanced in the presenceof fibrin mimic However, the N-terminal fragment isolated fromthis hybrid molecule (consisting of growth factor domain andkringle 2 domain) did interact with lysyl-Sepharose, suggestingthat in the hybrid molecule a functional lysyl-binding siteis present but not operational. Indeed, lysine analogue (e-amino-caproicacid) sensitive binding of isolated t-PA kringle 2 domain tou-PA could be observed. The modified u-PA kringle, the wildtype u-PA kringle and the kringle 2 of the u-PA hybrid werealso placed N-terminal of the protease domain of t-PA. As expected,the t-PA mutant consisting of the kringle 2 domain and the proteasedomain bound to lysyl-Sepharose and showed fibrin-dependentplasminogen activation. Further, the hybrid molecule consistingof the u-PA kringle placed N-terminal of the t-PA protease domaindid not display these features. Introduction of the modifiedu-PA kringle N-terminal of the t-PA protease domain resultedin a very weak interaction with lysyl-Sepharose. Despite thehigh overall similarity in primary structure of the modifiedu-PA kringle and t-PA kringle 2 (68%), no fibrin-dependent plasminogenactivation of this hybrid molecule was observed. The above-mentionedresults question the concept that the structural auto-nomousdomains within hybrid plasminogen activators t-PA and u-PA functionas autonomous domains and suggest that interactions betweenthe kringle and the protease domain in hybrid molecules stronglyinfluences their functional features 相似文献
5.
目的 评价尿激酶(UK)联用硫酸镁(MS)治疗急性心肌梗塞(AMI)的临床疗效及安全性。方法 151 例AMI 住院患者随机分为UK +MS 组(UM 组, 76 例)和UK 组(75 例), 除UM 组在静滴UK 前15 min 开始加用MS 外, 余相同。结果 UM 组的再通率有高于UK 组趋势, 而5 周病死率、心力衰竭、梗塞后心绞痛、再梗塞及梗塞延展的发生率均明显低于UK 组(P 均<0.01)。结论 UK 联用MS 治疗AMI 优于UK 疗法, MS 可作为一种安全有效的溶栓治疗辅助剂。 相似文献
6.
He—Ne激光血管内照射与尿激酶治疗脑梗塞对比研究及微观机理初探 总被引:8,自引:3,他引:5
将62例脑梗塞患者随机分为两组,并分别用激光血管内照射(ILIB)及尿激酶进行治疗,继而对两组疗效进行对比性观察、测试及评估。文末对ILIB的作用机理进行了分析。 相似文献
7.
Roles of the Na+/H+ Exchanger Isoform 1 and Urokinase in Prostate Cancer Cell Migration and Invasion
Xiuju Li Benjamin Buckley Konstantin Stoletov Yang Jing Marie Ranson John D. Lewis Mike Kelso Larry Fliegel 《International journal of molecular sciences》2021,22(24)
Prostate cancer is a leading cause of cancer-associated deaths in men over 60 years of age. Most patients are killed by tumor metastasis. Recent evidence has implicated a role of the tumor microenvironment and urokinase plasminogen activator (uPA) in cancer cell migration, invasion, and metastasis. Here, we examine the role of the Na+/H+ exchanger isoform 1 (NHE1) and uPA in DU 145 prostate cancer cell migration and colony formation. Knockout of NHE1 reduced cell migration. The effects of a series of novel NHE1/uPA hexamethylene-amiloride-based inhibitors with varying efficacy towards NHE1 and uPA were examined on prostate cancer cells. Inhibition of NHE1—alone, or with inhibitors combining NHE1 or uPA inhibition—generally did not prevent prostate cancer cell migration. However, uPA inhibition—but not NHE1 inhibition—prevented anchorage-dependent colony formation. Application of inhibitors at concentrations that only saturate uPA inhibition decreased tumor invasion in vivo. The results suggest that while knockout of NHE1 affects cell migration, these effects are not due to NHE1-dependent proton translocation. Additionally, while neither NHE1 nor uPA activity was critical in cell migration, only uPA activity appeared to be critical in anchorage-dependent colony formation of DU 145 prostate cancer cells and invasion in vivo. 相似文献
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9.
特发性肺间质纤维化(IPF)是一种进行性间质性肺疾病,其预后比许多癌症都要差,发病机制复杂并且尚未明确。在肺纤维化的动物模型以及IPF的患者中,已证实均存在凝血系统以及纤溶系统的失衡。本文阐述了凝血纤溶系统失衡在IPF中的可能损伤机制,为IPF的治疗提供新思路。 相似文献
10.
Polypropylene films with various amine groups were prepared by radiation‐induced grafting of glycidyl methacrylate (GMA) onto polypropylene (PP) films and the subsequent amination of poly‐GMA graft chains were examined. The physical and chemical properties of the GMA‐grafted PP film and the aminated PP film were investigated by IR, SEM, XPS, and TGA. The adsorption of urokinase for the PP films modified with various amine groups was examined under various conditions of different amine group contents and pH values. The adsorption of urokinase increased by increasing the amine group content. In the range of amine group content from 1.0 to 2.7 mmol/g, the adsorption of urokinase by the PP films modified with six kinds of amines was in the following order: trimethylamine > diethylenetriamine > triethylenetetraamine > ethylenediamine > dimethylamine > diethyleneamine. The adsorption amounts of urokinase by the PP film with the trimethylamine and triethylenetetraamine groups at pH 7.4 were higher than those at pH 9. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 80: 2851–2858, 2001 相似文献