胸膜腔内注入纤溶剂治疗渗出性胸膜炎的研究进展

纤维蛋白溶解剂胸膜腔注入辅助治疗渗出性胸膜炎在临床上较大规模应用已有20年历史,最近5年之前的主要文献均肯定了其疗效,认为其可以促进引流,减少外科手术十预率,缩短住院时间及降低费用,但2005年英国牛津的一个迄今最大规模的胸膜腔注入链激酶治疗感染性胸水的前瞻性双盲对照试验则否定了这一结论,目前这一疗法的临床前景仍有待明确.一些新型纤溶剂及辅助引流胸水药物的研究已取得一些进展,但仍未见应用于人体,需要试验来验证其治疗作用.我国的研究应用主要是用尿激酶治疗结核性胸水.但只局限于临床指标的观察,尚缺乏检测应用尿激酶后胸水纤维蛋白、流变性等性状指标改变的实验数据支持研究结果.

CLARITY-TIMI 28和COMMIT/CCS-2研究简介

动脉粥样硬化血栓栓塞所导致的疾病是世界上最大的致命性杀手.由赛诺菲-安万特集团出资,在全球开展了两项研究CLARITY-TIMI 28(CLopidogrel as Adjunctive ReperfusIon Therapy-Thrombolysis In Myocardial Infarction Study 28)和COMMIT/CCS-2(C1Opidogrel and Metoprolol in Myocardial Infarction Trial),这项两项重要临床研究结果在2005年3月第54届美国心脏病学会(ACC)年会上公布.应法国赛诺菲-安万特集团中国公司的邀请,著名心血管专家胡大一教授和Braunwald博士2005年3月25日在北京对有关媒体通报ACC年会上发表的CLARITY-TIMI28和COMMIT/CCS-2重要临床研究结果进行通报,结果显示:抗血小板药物氯吡格雷(75mg每日1次)联合标准治疗对于急性心肌梗死(AMI)[(ST段抬高AMI(STE-AMI)]的患者显著有益.两项重大临床研究共包括了近50000名患者.

急性心肌梗塞溶栓疗法并发症的护理

急性心肌梗塞是由于冠状动脉急性闭塞,使部分心肌严重持久的缺血而发生局部坏死,病情险恶,如果处理得当能延缓或避免发展到最终结果。溶栓治疗的作用机理是使冠状动脉重新开放和恢复心肌的正常血流。我科应用纤溶酶激活剂尿激酶(uk)

外科治疗脑室出血的现状

脑室出血是临床上常见的急危重病,具有病情重、变化快、病死率高特点;大多数需要外科治疗;本文从病理基础、手术时机、手术方式、常见纤溶剂、脑脊液置换等方面,对脑室出血外科治疗现状进行综述。

High level of urokinase plasminogen activator contributes to cholangiocarcinoma invasion and metastasis

AIM: To investigate the role of urokinase plasminogen activator (uPA) in cholangiocarcinoma (CCA) invasion and its correlation with clinicopathological parameters. METHODS: uPA expression in CCA tissue was determined by immunohistochemistry. The level of uPA from two CCA cell lines (HuCCA-1 and KKU-M213) and a noncancer immortalized cholangiocyte cell line (H69) was monitored by plasminogen-gelatin zymography and western blotting, whereas that of plasminogen activator inhibitor type 1 (PAI-1) protein and uPA receptor (uPAR)mRNA was monitored by western blotting and quantitative real-time reverse transcriptase polymerase chain reaction, respectively. Two independent methods were employed to suppress uPA function: a synthetic uPA inhibitor (B428) and silencing of uPA gene expression using siRNA. In vitro invasion of the uPA-disrupted cells was assessed by Matrigel-coated Transwell assay. RESULTS: The immunohistochemical study showed that 75.3% (131/174) of CCA tissues expressed uPA. High uPA expression was correlated with lymphatic invasion and metastasis of CCA patients. Plasminogen-gelatin zymography of the conditioned media and cell-surface eluates showed that both CCA cell lines, but not H69, expressed both secreted and membrane-bound forms of uPA. Although the two CCA cell lines, HuCCA-1 and KKU-M213, expressed a relatively high level of uPA and uPAR, the latter exhibited a much lower degree of in vitro invasiveness, correlating with a high expression of PAI-1 in the latter, but not in the former. Suppressing uPA function with a specific uPA inhibitor, B428, or with siRNA against uPA reduced in vitro invasiveness of KKU-M213 cells, demonstrating the requirement for uPA in the invasiveness of CCA cells. Therefore, our in vivo and in vitro studies suggest that uPA is an important requirement for the invasion process of CCA. CONCLUSION: uPA expression correlates with lymphatic invasion and metastasis in vivo and is required for CCA cell invasion in vitro , suggesting its potential as a therapeutic target.

INVESTIGATION OF THROMBOMODULIN AND PLASMINOGEN ACTIVATOR INHIBITOR TYPE-I IN PREGNANCY INDUCED HYPERTENSION AND ITS CLINICAL SIGNIFICANCE

Objective. To measure the circulating levels of thrombomodulin (TM) and plasminogen activator inhibitor type- I (PAI- I) in women with pregnancy induced hypertension (PIH). Methods. Blood samples were drawn from 97 pregnant women in their third trimester, grouped as 25 mild PIH,26 moderate PIH,22 severe PIH and 24 normotensive healthy pregnant women for determining levels of TM by ELISA,PAI- I by colorimetric assay methods, and creatinine (Cr) in serum by biochemical method. Results. Circulating levels of TM, PAI- I and TM/Cr ratio increased with increasing severity of PIH. There were no significant differences between mild and normotensive pregnant women. The parameters were significantly changed in the moderate and severe PIH groups. Conclusion. TM and PAI- I may serve as meaningful clinical markers for the assessment of the endothelial damage in PIH, which is very important in evaluating and following the development of PIH.

Over-expression of uPA increases risk of liver injury in pAAV-HBV transfected mice

AIM:To investigate the relationship between overexpression of urokinase plasminogen activator(uPA) and hepatitis B virus(HBV) related liver diseases in a transgenic mouse model.METHODS:Albumin-tetracycline reverse transcriptional activator and tetO-uPA transgenic mice were generated respectively through pronuclear injection and crossed to produce the double transgenic in-alb-uPA mice,for which doxycycline(Dox)-inducible and liver-specific over-expression of uPA can be achieved.Hydrodynamic transfection of plasmid adeno-associated virus(AAV)1.3HBV was performed through the tail veins of the Dox-induced in-alb-uPA mice.Expression of uPA and HBV antigens were analyzed through double-staining immunohistochemical assay.Cytokine production was detected by enzyme linked immunosorbent assay and α-fetoprotein(AFP) mRNA level was evaluated through real-time quantitative polymerase chain reaction.RESULTS:Plasmid AAV-1.3HBV hydrodynamic transfection in Dox-induced transgenic mice not only resulted in severe liver injury with hepatocarcinoma-like histological changes and hepatic AFP production,but also showed an increased serum level of HBV antigens and cytokines like interleukin-6 and tumor necrosis factor-α,compared with the control group.CONCLUSION:Over-expression of uPA plays a synergistic role in the development of liver injury,inflammation and regeneration during acute HBV infection.

A novel animal model for in vivo study of liver cancer metastasis

AIM:To establish an animal model with human hepatocyte-repopulated liver for the study of liver cancer metastasis.METHODS:Cell transplantation into mouse livers was conducted using alpha-fetoprotein(AFP)-producing hu-man gastric cancer cells(h-GCCs) and h-hepatocytes as donor cells in a transgenic mouse line expressing urokinase-type plasminogen activator(uPA) driven by the albumin enhancer/promoter crossed with a severe combined immunodeficient(SCID) mouse line(uPA/SCID mice).Host mice were divided into two groups(A and B).Group A mice were transplanted with h-GCCs alone,and group B mice were transplanted with h-GCCs and h-hepatocytes together.The replacement index(RI),which is the ratio of transplanted h-GCCs and h-hepatocytes that occupy the examined area of a histological section,was estimated by measuring h-AFP and h-albumin concentrations in sera,respectively,as well as by immunohistochemical analyses of h-AFP and human cytokeratin 18 in histological sections.RESULTS:The h-GCCs successfully engrafted,repopulated,and colonized the livers of mice in group A(RI = 22.0% ± 2.6%).These mice had moderately differentiated adenocarcinomatous lesions with disrupted glandular structures,which is a characteristics feature of gastric cancers.The serum h-AFP level reached 211.0 ± 142.2 g/mL(range,7.1-324.2 g/mL).In group B mice,the h-GCCs and h-hepatocytes independently engrafted,repopulated the host liver,and developed colonies(RI = 12.0% ± 6.8% and 66.0% ± 12.3%,respectively).h-GCC colonies also showed typical adenocarcinomatous glandular structures around the h-hepatocyte-colonies.These mice survived for the full 56 day-study and did not exhibit any metastasis of h-GCCs in the extrahepatic regions during the observational period.The mice with an h-hepatocyte-repopulated liver possessed metastasized h-GCCs and therefore could be a useful humanized liver animal model for studying liver cancer metastasis in vivo.CONCLUSION:A novel animal model of human liver cancer metastasis was established using the uPA/SCID mouse line.This model could be useful for in vivo testing of anti-cancer drugs and for studying the mechanisms of human liver cancer metastasis.

Synergistic action of non-solvent induced phase separation in preparation of poly(vinyl butyral) hollow fiber membrane via thermally induced phase separation

A systematic study of air gap distance effects on the structure and properties of poly(vinyl butyral)hollow fiber membrane via thermally induced phase separation(TIPS)has been carried out.The results show that the hollow fiber membrane prepared at air gap zero has no skin layer; the pore size near the outer surface is larger than that near the inner surface; and the special pore channel-like structure near the outer surface is formed,which is quite different with the typical sponge-like structure caused by TIPS and the finger-like structure caused by non-solvent induced phase separation(NIPS),because of the synergistic action of non-solvent induced phase separation at air gap zero.The pore size gradually decreases from outer surface layer to the intermediate layer,but increases gradually from intermediate layer to the inner surface layer.With the increase of air gap distance,the pore size near the outer surface gets smaller and a dense skin layer is formed,and the pore size gradually increases from the outer surface layer to the inner surface layer.Water permeability of the hollow fiber membrane decreases with air gap distance,the water permeability decreases sharply from 45.50×10-7 to 4.52×10-7 m3/(m2·s·kPa)as air gap increases from 0 to 10 mm at take-up speed of 0.236 m/s,further decreases from 4.52×10-7 to 1.00×10-8 m3/(m2·s·kPa)as the air gap increases from 10 to 40 mm.Both the breaking strength and the elongation increase with the increase of air gap distance.The breaking strength increases from 2.25 MPa to 4.19 MPa and the elongation increases from 33.9% to 132.6% as air gap increases from 0 mm to 40 mm at take-up speed 0.236 m/s.

THE INCREASE IN PLASMINOGEN ACTIVATOR INHIBITOR TYPE-1 EXPRESSION BY STIMULATION OF ACTIVATORS FOR PEROXISOME PROLIFERATOR-ACTIVATED RECEPTORS IN HUMAN ENDOTHELIAL CELLS

Objective.To investigate the effect of peroxis ome proliferator-activated recept ors(PPARs )activators on plasminogen activator inhibitor ty pe-1(PAI-1)expression in human umbilical vein e ndothelial cells and the possi-ble mechanism.Methods.Human umbilical vein endothelial ce lls(HUVECs )were obtained from normal fetus,and cul-tured conventionally.Then the HUVECs were exposed to test agents(linolenic acid,linoleic acid,oleic acid,stearic acid and prostaglandin J 2 respectively)in varying concentrations with fresh media.RT -PCR and ELISA were applied to determine the expression of PPARs and PAI-1in HUVECs.Results.PPARα,PPARδand PPARγmRNA were detected by using RT-PCR in HUVECs.Treatment of HUVECs with PPARαand PPARγactivators---linolenic acid,linoleic acid,oleic acid and prostaglandin J 2 respectively,but not with stearic a cid could augment PAI-I mRNA expression and protein secretion in a concentration-dependent manner.However,the mRNA expressions of 3subclasses of PPAR with their activators in HUVECs were not changed compared w ith controls.Conclusion.HUVECs express PPARs.PPARs activators may increase PAI-1expression in ECs,but the underlying mechanism remains uncle ar.Although PPARs expression was not enhanced after stimulated by their activators in ECs,the role of functionally active PPARs in regulating PA I-1expression in ECs needs to be further investigated by using transient gen e transfection assay.

Cloning, Expression and Activity Analysis of a Novel Fibrinolytic Serine Protease from Arenicola cristata

The full-length c DNA of a protease gene from a marine annelid Arenicola cristata was amplified through rapid amplification of c DNA ends technique and sequenced. The size of the c DNA was 936 bp in length, including an open reading frame encoding a polypeptide of 270 amino acid residues. The deduced amino acid sequnce consisted of pro- and mature sequences. The protease belonged to the serine protease family because it contained the highly conserved sequence GDSGGP. This protease was novel as it showed a low amino acid sequence similarity(< 40%) to other serine proteases. The gene encoding the active form of A. cristata serine protease was cloned and expressed in E. coli. Purified recombinant protease in a supernatant could dissolve an artificial fibrin plate with plasminogen-rich fibrin, whereas the plasminogen-free fibrin showed no clear zone caused by hydrolysis. This result suggested that the recombinant protease showed an indirect fibrinolytic activity of dissolving fibrin, and was probably a plasminogen activator. A rat model with venous thrombosis was established to demonstrate that the recombinant protease could also hydrolyze blood clot in vivo. Therefore, this recombinant protease may be used as a thrombolytic agent for thrombosis treatment. To our knowledge, this study is the first of reporting the fibrinolytic serine protease gene in A. cristata.

Novel distribution pattern of fibrinolytic components in rabbit tissues extract: a preliminary study

Objective: The purpose of this work was to investigate the distribution pattern of fibrinolytic factors and their inhibitors in rabbit tissues. Methods: The components of the fibrinolytic system in extracts from a variety of rabbit tissues, including tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), plasminogen (Plg), plasmin (Pl) and α2 plasmin inhibitor (α2PI), were determined by colorimetric assay. Results: The tissue extracts in renal, small intestine, lung, brain and spleen demonstrated strong fibrinolytic function, in which high activity of tPA, Plg and Pl was manifested; whereas in skeletal muscle, tongue and stomach, higher activity of PAI-1 and α2PI showed obviously. Also excellent linear correlations were found between levels of tPA and PAI-1, Pl and α2PI, Plg and Pl. In related tissues, renal cortex and renal marrow showed distinctly higher activity of tPA and lower activity of PAI-1, with the levels of Plg and Pl in renal cortex being higher than those in renal marrow, where the α2PI level was higher than that in renal cortex. Similarly, the levels of tPA, Plg and Pl in small intestine were higher than those in large intestine, but with respect to PAI-1 and α2PI, the matter was reverse. In addition, the fibrinolytic activity in muscle tissue was lower, however, the levels of tPA, Plg, and Pl in cardiac muscle were obviously higher than those in skeletal muscles, and the levels of PAI-1 and α2PI were significantly lower than those in skeletal muscle. Conclusion: Our data demonstrate that a remarkable difference of the fibrinolytic patterns exists in rabbit tissues, which has probable profound significance in understanding the relationship between the function of haemostasis or thrombosis and the physiologic function in tissues.

Effect of carboxymethyl cellulose on dissolution kinetics of carboxymethyl cellulose-sodium carbonate two-component tablet

Sodium carbonate and carboxymethyl cellulose powders are compressed into two-component tablets with three mass ratios,97%:3%,95%:5% and 93%:7%.The dissolution tests for two-component tablets and reference pure sodium carbonate tablets are carried out at various temperatures.The dissolution process of each tablet is measured by electrical conductivity tracking method and the concentration of dissolved sodium carbonate is quanti fied with calibrated conductivity-concentration converting equation of sodium carbonate.The quanti fied dissolution data is fitted with both surface reaction model and diffusion layer model and the results clearly show that surface reaction model is suggested as the appropriate dissolution model for all measured tablets.Therefore,it is determined that carboxymethyl cellulose is a stable element to remain the dissolution mechanism of tablet unchanged.The dissolution rate constant quanti fied with surface reaction model presents that carboxymethyl cellulose-sodium carbonate two-component tablets obtain signi ficant higher dissolution rate constant than pure sodium carbonate tablet and higher proportion of carboxymethyl cellulose leads to apparent higher dissolution rate constant.The results prove for the usage of carboxymethyl cellulose in most practical applications at a relative low-level,the effect of carboxymethyl cellulose is effective and positive for two-component tablet to enhance the dissolution process and improve dissolution rate constant and this effect is speculated coming from its dynamic physical transforming process in water including dilation and conglutination.

关于联合使用纤维蛋白溶解剂/脱氧核糖核酸酶治疗胸腔感染的建议

就在十余年前,人们还普遍认为胸腔内注入纤维蛋白溶解剂(纤溶剂)治疗胸腔感染是无效的。随着临床研究证据的逐渐积累,现在国际上已经达成了共识,推荐以胸腔内同时注入纤溶剂和脱氧核糖核酸酶作为胸腔感染的初治用药,或作为外科手术后的后续治疗方案。推荐使用剂量为:组织纤维蛋白溶酶原激活剂10 mg/次,2次/d;脱氧核糖核酸酶5 mg/次,2次/d。建议今后的研究重点将在于摸索出更优化的用药方案,并开发更有效的治疗药物。