Study of the fibrinolytic activity of serrapeptase and its in vitro thrombolytic effects

Abstract Serrapeptase, a proteolytic enzyme, has been used for the adjuvant treatment of many diseases. However, its fibrinolytic activity is still uncertain. Herein, the fibrinolytic activity of serrapeptase and its in vitro thrombolytic effects were investigated. The results showed that the fibrinolytic activity of serrapeptase was 1295 U/mg, and the specific activity was 3867 U/mg of protein when its proteolytic activity toward casein was 2800 U/mg. The optimum temperature and pH for serrapeptase activity were 37-40°C and 9.0, respectively. At 1 mmol/L, Zn2+, Mn2+ and Fe2+ could activate the fibrinolytic activity of serrapeptase, while K+, Cu2+, sodium dodecyl sulfate (SDS) and ethylene diamine tetraacetic acid (EDTA) inhibited it. In vitro tests showed that serrapeptase could completely prevent blood coagulation at 150 U/mL, and the percentage of blood clot lysis reached 96.6% at 37°C after 4 h at 300 U/mL. These results indicate that serrapeptase has excellent fibrinolytic activity, and can be used as a health food or candidate drug for the prevention or treatment of thrombotic diseases.

Study on influencing factors of plasma FDP and D-dimer levels of newborns in NICU / 国际儿科学杂志

Objective:To investigate the influencing factors of physiological and pathological conditions at birth of newborn and gestational conditions of pregnant mothers on plasma fibrin/fibrinogen degradation products(FDP)and D-dimer levels.Methods:From May 1, 2018 to October 31, 2018, 222 newborns admitted to NICU of the Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology were enrolled in this study.Newborns were sent to NICU within 2 hours after birth and venous blood was collected immediately.The levels of FDP and D-dimer were detected by immunoturbidimetry.Groups were divided according to different gender, gestational age, birth weight, relationship between gestational age and birth weight, mode of delivery, asphyxia at birth, acidosis, antenatal hormone use, anticoagulant drugs, and perinatal risk factors(gestational hypertension, premature rupture of membranes, abnormal placenta, gestational diabetes). The levels of plasma FDP and D-dimer were compared among the groups.Mann Whitney U test, Kruskal Wallis H test, Spearman rank correlation and generalized linear model were used for statistical analysis. Results:The plasma FDP and D-dimer values of 222 NICU neonates were skewed at birth, with median values of 6.00 mg/L and 1.74 mg/L, and quartile distances of 10.40 mg/L and 2.55 mg/L, respectively.The concentrations of FDP in neonates born to natural labour and cesarean section were 11.70 mg/L and 5.30 mg/L, respectively, and D-dimer concentrations were 2.92 mg/L and 1.52 mg/L, respectively.The FDP and D-dimer levels were significantly higher in the former(Z values were -4.006 and -4.198, respectively, P<0.05). The levels of FDP and D-dimer in newborns with different gestational age, different birth weight and different blood pH values were compared respectively, and the differences were statistically significant( χ2 values were 15.322, 18.394, 9.677, 11.492, 7.023 and 8.345, respectively, P<0.05). Further analysis showed that the levels of FDP and D-dimer in neonates with gestational age < 34 weeks were significantly higher than those in~<37 weeks group and ≥37 weeks group( P<0.05). The FDP and D-dimer levels in the birth weight<1 500 g group were significantly higher than those in~<2 500 g group and ≥2 500 g group( P<0.05). Higher FDP and D-dimer levels were found in the pH<7.20 group than in the pH ≥7.35 group( P<0.05). A generalized linear model was established for multifactor analysis.The results showed that the concentration of FDP and D-dimer in plasma was related to gestational age, birth weight and arterial pH value. Conclusion:The levels of plasma FDP and D-dimer in NICU newborns at birth were influenced by gestational age, birth weight and acid-base balance.

Potential of modulating fibrinolytic activity in intervention of transfusion-related acute lung injury / 中国输血杂志

Transfusion-related acute lung injury (TRALI), with clinical manifestation, diagnosis and pathological mechanism consistent with acute lung injury(ALI), belongs to a sub-category of ALI. Excessive deposition of fibrin in lung is one of the characteristic of ALI, and reversing fibrin formation is of great significance to intervene ALI. The decrease of fibrinolytic activity is one of the important causes of excessive deposition of fibrin in lung, and also the important pathological feature of TRALI. This article discusses the potential of modulating fibrinolytic activity to intervene TRALI from the perspective of regulating the effectiveness of fibrinolytic activity to intervene ALI.

Rapid Enrichment and Activity Research of Antithrombotic Proteins from Pheretima vulgaris / 中国实验方剂学杂志

Objective:A strong antithrombotic protein component， named PvQ， was purified and enriched from total protein of <italic>Pheretima vulgaris</italic>，<italic> </italic>a<italic> </italic>traditional Chinese medicine. Moreover， we evaluated its fibrinolytic and anticoagulant activity， and expected to provide reference for the research on antithrombotic substances of Pheretima. Method:A rapid <italic>in</italic> <italic>vitro</italic> activity-oriented separation combined with the AKTA-Pure protein purification system conducted on <italic>P. vulgaris</italic>. Meanwhile， the fibrinolytic and anticoagulant activities of PvQ were measured by fibrin plate method and fibrinogen-thrombin time （Fibg-TT） method. And the <italic>in vitro</italic> thrombolysis assay was used for evaluating the lysis ability of PvQ to thrombus. Then the stability of PvQ was also analyzed for its anticoagulant activity at different pH and temperature. Result:The PvQ was successfully enriched and its activity was determined to have significant fibrinolytic and anticoagulant activities. And the result of <italic>in vitro</italic> thrombolysis assay revealed that PvQ could hydrolyze more than 80% of thrombus after 5 h of incubation at 37 ℃. In addition， the changes of temperature and pH had significant effects on antithrombotic activity， and this study showed that PvQ was rapidly inactivated at ≥60 ℃ or in acidic conditions （pH<7）. While， the activity of PvQ was unaffected or less affected at ≤50 ℃ and under alkaline conditions. Conclusion:A feasible preparation method of PvQ is established， and it can affect fibrin and fibrinogen at the same time， thus exerting a dual fibrinolytic effect and possessing significant fibrinolytic and anticoagulant activities. It provides a scientific interpretation for the treatment of thrombotic diseases by PvQ and a reference for the development of antithrombotic protein products of Pheretima.

Study on correlation between postoperative coagulation function and fibrinolytic activity indexes with deep vein thrombosis in patients with traumatic fracture / 国际检验医学杂志

Objective To investigate the correlation between the coagulation function and fibrinolytic activity with deep venous thrombosis (DVT) after traumatic fracture operation.Methods One hundred and thirty-four cases of traumatic fracture from April 2010 to May 2014 were divided into the DVT group(24 cases) and non-DVT group,and at the same time 110 healthy people were selected as the control group.The venous blood at different time points was collected for observing the levels change of prothrombin time (PT),activated partial thromboplastin time (APTT),fibrinogen (FIB),platelet aggregation rate (PAgT),dipolymer (D-D),thrombin antithrombin complex (TAT),plasminogen activity (PIg) by using appropriate methods.Results There was no difference in APTT and PT groups among the groups(P>0.05).Preoperative FIB,PAgT,D-D and TAT levels was the DVT group >non-DVT group > control group,while the level of PIg was the DVT group0.05).Conclusion The increase of D-D and TAT is closely related with DVT in traumatic fracture and,which is an independent risk factor.

In vitro characterization of jellyfish venom fibrin(ogen)olytic enzymes from Nemopilema nomurai

Abstract Background: Because jellyfish are capable of provoking envenomation in humans, they are considered hazardous organisms. Although the effects of their toxins are a matter of concern, information on the venom components, biological activity and pathological mechanisms are still scarce. Therefore, the aim of the present study was to investigate a serine protease component of Nemopilema nomurai jellyfish venom (NnV) and unveil its characteristics. Methods: To determine the relationship between fibrinolytic activity of NnV and the serine protease, fibrin zymography was performed using metalloprotease and serine protease inhibitors. The biochemical characterization of serine proteases of NnV were determined by the amidolytic assay. Fractions with fibrinolytic activity were obtained by DEAE cation exchange column. Results: NnV displayed fibrinolytic activities with molecular masses of approximately 70, 35, 30, and 28 kDa. The fibrinolytic activity of NnV was completely obliterated by phenylmethylsulfonyl fluoride, a prototype serine protease inhibitor. Based on amidolytic assays using chromogenic substrates specific for various kinds of serine proteases, NnV predominantly manifested a chymotrypsin-like feature. Its activity was completely eliminated at low pH ( 6) and high temperatures (> 37 °C). Some metal ions (Co2+, Cu2+, Zn2+ and Ni2+) strongly suppressed its fibrinolytic activity, while others (Ca2+ and Mg2+) failed to do so. Isolation of a serine protease with fibrionolytic activity from NnV revealed that only p3 showed the fibrinolytic activity, which was completely inhibited by PMSF. Conclusion: The present study showed that N. nomurai jellyfish venom has a chymotrypsin-like serine protease with fibrinolytic activity. Such information might be useful for developing clinical management of jellyfish envenomation and pharmacological agents with therapeutic potential for thrombotic diseases in the future.

In vitro characterization of jellyfish venom fibrin(ogen)olytic enzymes from Nemopilema nomurai

Background: Because jellyfish are capable of provoking envenomation in humans, they are considered hazardous organisms. Although the effects of their toxins are a matter of concern, information on the venom components, biological activity and pathological mechanisms are still scarce. Therefore, the aim of the present study was to investigate a serine protease component of Nemopilema nomurai jellyfish venom (NnV) and unveil its characteristics. Methods: To determine the relationship between fibrinolytic activity of NnV and the serine protease, fibrin zymography was performed using metalloprotease and serine protease inhibitors. The biochemical characterization of serine proteases of NnV were determined by the amidolytic assay. Fractions with fibrinolytic activity were obtained by DEAE cation exchange column. Results: NnV displayed fibrinolytic activities with molecular masses of approximately 70, 35, 30, and 28 kDa. The fibrinolytic activity of NnV was completely obliterated by phenylmethylsulfonyl fluoride, a prototype serine protease inhibitor. Based on amidolytic assays using chromogenic substrates specific for various kinds of serine proteases, NnV predominantly manifested a chymotrypsin-like feature. Its activity was completely eliminated at low pH (< 6) and high temperatures (> 37 °C). Some metal ions (Co2+, Cu2+, Zn2+ and Ni2+) strongly suppressed its fibrinolytic activity, while others (Ca2+ and Mg2+) failed to do so. Isolation of a serine protease with fibrionolytic activity from NnV revealed that only p3 showed the fibrinolytic activity, which was completely inhibited by PMSF. Conclusion: The present study showed that N. nomurai jellyfish venom has a chymotrypsin-like serine protease with fibrinolytic activity. Such information might be useful for developing clinical management of jellyfish envenomation and pharmacological agents with therapeutic potential for thrombotic diseases in the future.(AU)

Effect of factors on pepsin and trypsin fibrinolytic activity and deactivation of fibrinolytic activity / 中草药

Objective: To investigate the method for study on the effect of factors on pepsin and trypsin fibrinolytic activity and deactivation of fibrinolytic activity and to eliminate the interference of pepsin and trypsin on the detection of crude protein fibrinolytic activity of Armadillidium vulgare (porcellio plasmin) in order to obtain the proteins or peptides which have the smaller molecular weight but higher titer during the pepsin and trypsin degradation. Methods: To study the effect of pepsin and trypsin deactivation on pH value, temperature, metal ions, enzyme inhibitor, surfactant, and responsing fibrinolytic by fiber fibrin plate assay. The better enzyme deactivation process was obtained and used for studying the effect on the fibrinolytic activity of urokinase, lumbrokinase, and porcellio plasmin. Results: All the pH value, temperature, metal ions, enzyme inhibitor, and surfactant have had an impact on pepsin and trypsin fibrinolytic activity. Among them the optimum deactivation of pepsin was pH 6.0-8.0, while the optimum deactivation of trypsin was mixed preparation with TLCK at the concentration of 25 mg/mL and EDTA at the concentration of 1 mmol/L. Conclusion: This study has obtained the better enzyme deactivation process which could be used for the detection of fibrinolytic activity of pepsin and trypsin degradation product by fiber fibrin plate assay, the operation is simple, and the repeatability and stability are good.

Caracterización bioquímica y biológica del veneno de la serpiente Neotropical Macagua (Bothrops Colombiensis) de la región de Barlovento, estado Miranda, Venezuela / Biological and biochemical characterization of the Neotropical Macagua (Bothrops colombiensis) snake venom from Barlovento region, Miranda state, Venezuela

Introducción: el veneno de B. colombiensis no es solamente un elemento tóxico; en su composición existen múltiples componentes, que tienen un gran potencial terapéutico, principalmente en el tratamiento de patologías de la trombosis y la coagulación. Objetivos: estudiar una mezcla de venenos de Bothrops colombiensis de una ubicacion geográfica de Venezuela, a fin de hacer un barrido de sus actividades hemostáticas, que permitirá posteriormente purificar y caracterizar moléculas con actividad antitrombótica y anticoagulantes, entre otras, con potencial terapéutico. Métodos: el veneno a estudiar, es una mezcla de ellos obtenidos de serpientes provenientes de la Región de Barlovento, estado Miranda, Venezuela. Se caracterizó bioquímicamente por cromatografias de exclusión molecular, cromatografía de fase reversa C18 y por electroforesis a través de SDS­PAGE; y biológicamente por medio de actividades relacionadas con la hemostasia. Se analizaron los perfiles en relación a las actividades fibrinolítica, proteolítica sobre polvo azul y cadena ß de insulina, procoagulante, hemorrágica y letal. Resultados: la actividad hemorrágica, definida como la Dosis Hemorrágica Mínima fue de 8,7 mg/kg. La letalidad, definida como la Dosis Letal cincuenta fue 8,7 mg/kg. El veneno presentó actividad procoagulante y fibrinolítica. Las fracciones mostraron actividad fibrinolítica y proteolítica sobre polvo azul de ocultamiento y sobre la cadena ß de insulina. Conclusiones: las características biológicas de los componentes de este veneno le confieren un enorme potencial terapéutico, ya que contiene una alta actividad fibrinolítica y anticoagulante. Estos compuestos una vez purificados y caracterizados podrían explorarse como coadyuvantes en procesos trombolíticos, dado que disuelven coágulos de fibrina y degradan fibrinógeno, evitando episodios de retrombosis(AU)

Introduction: This paper is a screening of multiple toxic activities, of which some will be potentially useful for the management of coagulation pathologies. Objetives: A pool of Bothrops colombiensis venoms from a specific geographical location was studied, in order to carry out a hemostatic activities screening, allowing then to purify and characterise molecules with antithrombotic and anticoagulant activity, among others, which could have therapeutic potential. Methods: The venom was chromatographically by molecular exclusion and reverse phase C18 and SDS -PAGE characterized; its hemostatic activity was also established. Snakes were from the region of Barlovento, Miranda state, Venezuela. Profiles of fibrinolytic, proteolytic, procoagulant, hemorrhagic and lethal activities were analyzed. Hemorrhagic activity was 8.7 mg/kg. The LD50 was 8.7 mg/kg. The venom showed strongly procoagulant activity. Both, crude venom as fractions showed high fibrinolytic activity. The majority of the eluted fractions showed significant proteolytic activity in azure blue powder and on ß chain of insulin. Conclusions: The biological characteristics of the components of this venom confer enormous therapeutic potential because they contain a high fibrinolytic and anticoagulant activity. Most of these proteinases, once purified and characterized, could be explored as thrombolytic agents given that dissolves fibrin clots or prevent their formation(AU)

Effect of Compound Sanggou Granules on the Fibrinogen, Antithrombin Ⅲ and Fibrinolytic Activity in Hyperlipidemia Rats / 中国药师

Objective:To investigate the effect of compound Sanggou granules on the activity of Fib, AT-Ⅲ, t-PA, PAI-1 and t-PA/ PAI-1 in hyperlipidemic rats. Methods: The hyperlipidemic rat model was established by feeding high fat diet to SD male rats. Sixty healthy SD rats were randomly divided into five groups: the normal diet control group, high fat control group, high dose drug group, low dose drug group and fluvastatin sodium group. Four weeks after the administration, the blood samples were withdrawn for the determination of the levels of blood lipid, Fib, A-Ⅲ, t-PA, PAI-1 and t-PA/ PAI-1. Results:Compared with those of the normal diet control group, the levels of TC, LDL-C, Fib and PAI-1 were increased and the levels of HDL-C, t-PA , AT-Ⅲand t-PA/ PAI-1 were decreased significantly (P<0. 01) in the high fat control group. Compared with those of the high fat control group, the levels of TC, LDL-C, PAI-1 and Fib were decreased(P<0. 01 or P<0. 05),and the levels of HDL-C, t-PA AT-Ⅲ and t-PA/PAI-1 were in-creased significantly in the high dose drug group (P<0. 05 or P<0. 01). The similar effects were shown in the fluvastatin sodium group with the stability of AT-Ⅲ. The levels of TC, LDL-C and PAI-1 were decreased and the levels of t-PA/PAI-1 were increased no-tably in the low dose drug group. Conclusion: Compound Sanggou granules exhibit hypolipidemic effect in hyperlipidemic rats, and can improve hypercoagulability and enhance anticoagulation and fibrinolytic activity in hyperlipidemic rats. Furthermore, compound Sanggou granules at high dose show the same effect as fluvastatin sodium, even in anticoagulation, the granules are superior to fluvasta-tin sodium.

Platelet aggregability and fibrinolytic activity in different phases of menstrual cycle.

Background:- Sexual dimorphism in coronary artery disease (CAD) mortality is attributed to the cardioprotective effects of estrogen.This is reinforced by the observation that incidence of myocardial infarction is higher in menstrual phase, corresponding with low estrogen levels, in people who are predisposed to CAD due to the presence of modifiable risk factors. Cyclical variability of estrogen and progesterone in normal menstruating women may be associated with variability of platelet aggregation and fibrinolytic activity.There exists a delicate balance between fibrinolytic activity and platelet aggregation governing the haemostatic status. Objectives:- Platelet aggregability and fibrinolytic activity were measured and compared during menstrual (1-5 days), follicular ( 9-12 days) and luteal (20-25days) phases of menstrual cycle. Method:- In this cross sectional study of 50 normal menstruating females in age group of 18-35yrs, Platelet aggregability was measured by ADP induced platelet aggregation on a spectrophotometer. Fibrinolytic activity was estimated by euglobulin clot lysis time. Results :- Results were analyzed by students unpaired ‘t’ test. Change in platelet aggregability was found 0.12 ± 0.15, 0.04 ± 0.04 and 0.08 ± 0.07 in menstrual, follicular and luteal phase respectively. Platelet aggregability was found significantly (p < 0.001) higher in menstrual and luteal phases than follicular phase. The mean euglobulin clot lysis time was found 277.6 ± 43.96, 147.6 ± 52.78 and 244.6 ± 59.12 in menstrual, follicular and luteal phase respectively. Fibrinolytic activity was found significantly (p < 0.0001) lower in menstrual and luteal phases than follicular phase. Conclusion :- According to the present study, in both luteal and menstrual phases, not only platelet aggregability was found higher, but fibrinolytic activity was also found lower as compared to follicular phase, thereby pointing towards thrombotic tendency in these phases. Hence, these phases require careful monitoring in women who are susceptible to thrombotic disorders. However, follicular phase with lower platelet aggregability and higher fibrinolytic activity is relatively free from thrombotic risk.

Enhancement of protease production by Pseudomonas aeruginosa isolated from dairy effluent sludge and determination of its fibrinolytic potential

Objective: The present study aimed at isolating proteolytic bacteria from dairy effluent sludge, designing the process parameters for the enhanced production of protease and determination of its fibrinolytic potential. Methods: The dairy sludge was processed according to the microbiological criteria for the isolation of proteolytic bacteria. All the isolates were screened for their protease production ability and the isolate showing highest proteolysis was selected for further studies. Effects of various media components and process parameters like carbon and nitrogen supplementation, temperature, pH and incubation period were investigated. Partial purification of the protease was done using ammonium sulphate fractionation, following which its molecular weight and fibrinolytic activity were determined. Results: Based on the biochemical studies, the selected isolate was identified as Pseudomonas aeruginosa. The highest protease yield was obtained with maltose and yeast extract as supplements. The optimum pH, temperature and incubation period for protease production by the isolate was found to be 7.0, 37℃ and 48 h respectively. The partially purified enzyme preparation showed a single protein band in sodium dodecyl sulphate polyacrylamide gel electrophoresis, revealing the apparent molecular weight of the enzyme to be 35 kDa. The efficient removal of the blood stain emphasized its fibrinolytic potential. Conclusions: From the present study it is envisaged that cultural parameters significantly affect the protease production. Based upon the fibrinolytic activity, this protease may find broad applications in detergent and pharmaceutical industries.

Fibrinolysis and anticoagulant potential of a metallo protease produced by Bacillus subtilis K42.

In this study, a potent fibrinolytic enzyme-producing bacterium was isolated from soybean flour and identified as Bacillus subtilis K42 and assayed in vitro for its thrombolytic potential. The molecular weight of the purified enzyme was 20.5 kDa and purification increased its specific activity 390-fold with a recovery of 14%. Maximal activity was attained at a temperature of 40°C (stable up to 65°C) and pH of 9.4 (range: 6.5–10.5). The enzyme retained up to 80% of its original activity after pre-incubation for a month at 4°C with organic solvents such as diethyl ether (DE), toluene (TO), acetonitrile (AN), butanol (BU), ethyl acetate (EA), ethanol (ET), acetone (AC), methanol (ME), isopropanol (IP), diisopropyl fluorophosphate (DFP), tosyl-lysyl-chloromethylketose (TLCK), tosyl-phenylalanyl chloromethylketose (TPCK), phenylmethylsulfonylfluoride (PMSF) and soybean trypsin inhibitor (SBTI). Aprotinin had little effect on this activity. The presence of ethylene diaminetetraacetic acid (EDTA), a metal-chelating agent and two metallo protease inhibitors, 2,2′-bipyridine and o-phenanthroline, repressed the enzymatic activity significantly. This, however, could be restored by adding Co2+ to the medium. The clotting time of human blood serum in the presence of this enzyme reached a relative PTT of 241.7% with a 3.4-fold increase, suggesting that this enzyme could be an effective antithrombotic agent.

The association of urinary albumin excretion with fibrinolytic activity in patients with type 2 diabetes / 中国基层医药

Objective To examine the relationship between UAE and fibrinolytic activity in patients with type 2 diabetes.Methods 129 type 2 diabetic patients recruited and subgrouped by UAE,and the UAE,FBS,lipids,renal function and the activity of t-PA and PAI-1 were conducted in these patients and 40 health people.Results Compared to control group,the activity of PAI-1 was increased and the activity of t-PA decreased dramaticlly in type 2 diabetic group(P＜0.05);there was significant difference in the ratio of PAI-1 and t-PA between three diabetic groups(P＜0.05),of three group the ratio of PAI-1 and t-PA was most highest in the patients with macroal buminuria,and simple correlation analysis showed positive correlation between UAE and the ratio of PAI-1 and t-PA,especially in the patients with microalbuminuria(r=0.321,P＜0.05).Conclusion The UAE is closely relatedto fibrinolytic activity in patients type 2 diabetes.

Study on Effects of Saikosaponin-d on TPA,PAI,MDA and NO in Rats with Liver Fibrosis / 中国药房

OBJECTIVE:To investigate the effects of saikosaponin-d(SSd) on tissue plasminogen activator(TPA),plasminogen activator inhibitor(PAI),malonaldehyde(MDA) and NO in rats with liver fibrosis induced by dimethylnitrosamine(DMN).METHODS:Eighteen SD healthy rats were randomized to control group(NS ip qd for 4 weeks),model group(10mg? kg-1 DMN ip 3 times per week for 4 weeks) and SSd-treated group(10mg? kg-1 DMN ip 3 times per week + SSd 1.8 mg? kg-1 ip for 28 consecutive days).All rats were killed 1h after the last time of administration,blood samples were taken from abdominal aorta and liver samples were taken for the observation of pathology and detection of indices of TPA,PAI,MDA and NO etc.RESULTS:SSd could lessen the degree of liver fibrosis and improve the fibrinolytic activities of TPA and PAI,meanwhile,it showed clearance effect on MDA and marked protective effect on hepatic cells.There were significant differences between SSd-treated group and the model group.CONCLUSION:SSd exhibited protective function on experimental hepatic fibrosis in rats,which may be attributed to the improving of fibrinolytic activity,eliminating of lipid peroxidation and enhancing of NO level.

STUDIES ON FIBRINOLYTIC FUNCTION OF SUBTILISIN AND ITS MECHANISM / 营养学报

Objective:To evaluate the effect how Subtilisin FS33 affect thrombotic and fibrinolytic systems in vitro and in vivo. Method:Activity of Subtilisin FS33 was measured by clot liquefaction time(CLT) . On the model of 10% FeCl3 induced thrombi of carotid arteries in rats,various doses of Subtilisin FS33 were injected to the rats,and the fibrinolytic effect was observed. Results:0.5 g of the unheated blood clots gradually dissolved within 45 min,whereas the blood clots heated at 80℃ for 30 min dissolved within 3 h. This indicated that the enzyme was able to degrade blood clots in the absence of endogenous fibrinolytic factors. The experiment in vivo indicated that high dose subtilisn group could significantly prolong CT(coagulation time ) ,PT(prothrombin time) ,TT(thrombin time ) ,APTT(activated partial thromboplastin time) ,reduce ELT(euglobulin lysis time) ,decrease the content of FIB(fibrinogen) ,increase the content of FDP(fibrinogen degradation products) . D-dimer of all experimental groups waspositive. The venous thrombus in lung and kidney was dissolved totally or partly as observed by pathological section. Conclusion:Both thrombolytic effects of Subtilisin FS33 in vitro and in vivo were significant and the mechanisms might be associated with enhancing anticoagulation activity and fibrinolysis.

Fibrinolytic Activities and Their Relations to Esophageal Variceal Bleeding in Patients with Liver Cirrhosis / 대한소화기학회지

BACKGROUND/AIMS: Esophageal variceal bleeding in liver cirrhosis is a major complication and has high mortality rate. We tried to find fibrinolytic parameters, which correlated with variceal bleeding in cirrhotic patients. METHODS: We divided the cirrhotic patients into two groups: bleeding group (group A, n=15) and non-bleeding group (Group B, n=17). Fibrinolytic parameters (fibrinogen, D-dimer, plasminogen, tissue plasminogen activator [t-PA], fibrin degradation product [FDP], and plasminogen activator inhibitor type-1 [PAI-1]) were compared between two groups. In the group A, serial samplings were taken at the initial period, 3 days, 8 days, 15 days and 6 weeks after the bleeding onset. RESULTS: Plasma levels of FDP and D-dimer in the group A were significantly higher than the group B (1.7 +/- 1.16 vs. 0.95 +/- 1.27 mg/L and 10.96 +/- 6.58 vs. 4.99 +/- 3.50 micro gram/mL, respectively, p value<0.05). The clinical, biochemical, and coagulation parameters didn't show significant differences in both groups. The fibrinolytic parameters were improved along with the hemodynamic stabilization in group A. CONCLUSIONS: Cirrhotic patients with increased fibrinolytic activity were at higher risk of bleeding. Thus, the measurement of these parameters would be useful to identify patients at higher risk of esophageal variceal bleeding.

Effect of valsartan and amlodipine on platelet activation and fibrinolytic activity in elderly patients with essential hypertension / 中国临床药理学与治疗学

AIM: To observe platelet activation and fibrinolytic activity and evalute the effects of valsartan and amlodipine in elderly patients with essential hypertension. METHODS: Double antibody sandwich ELISA and spectrophotometric assay were used to examine the levels of platelet alpha granule membrane protein (GMP 140), tissue type plasminogen activator (t PA) and its inhibitor (PAI 1) in 57 elderly patients with essential hypertension and 30 normotensives. Valsartan was given in 29 cases (groupⅠ) and amlodipine in 28 cases (group Ⅱ) for 12 weeks. RESULTS: Elevated GMP 140, decreased t PA activity, and elevated PAI 1 activity were detected in the patients, but not in normotensives (P

Effects of 38.DEG.C. Bathing for 30 min on Hemostatic Function and Autonomic Nervous Function in Patients with Cerebral Infarction / 日本温泉気候物理医学会雑誌

Effects of 38°C 30-minute bathing on hemostatic function and autonomic nervous function were studied in 15 48-to-72-year-old patients with cerebral infarction. Blood samples were collected three times: immediately before the bathing, at the end of 30 minutes of bathing, and 30 minutes after the bathing. Hematocrit values and fibrinogen concentrations decreased during bathing and returned to the pre-bathing levels 30 minutes after bathing. This indicates that bathing caused hemodilution due to the fluid shift. During bathing, noradrenaline decreased at a rate significantly higher than that of hemodilution while the sympathetic nervous function, which was evaluated by spectral analysis of sequential variation in arterial blood pressure, was not suppressed. The autonomic nervous system seemed to be inactive in these patients. Coagulation time (PT and APTT) and platelet factor (β-TG and PF4) showed few changes. In the fibrinolytic system, however, tissue plasminogen activator (t-PA) antigen levels increased and plasminogen activator inhibitor type-1 (PAI-1) levels decreased after 30 minutes of bathing. This suggests that fibrinolytic activity was enhanced by 38°C bathing for 30 minutes. Thus, subthermal bathing with comfort may be useful in preventing cerebral infarction.

Separation, Purification and Analysis of the Components of Earthworm Fibrinoly tic Enzymes / 生物化学与生物物理进展

The earthworm fibrinolytic enzymes (EFE) were separated by affinity ch romatography using soybean trypsin inhibitor as a matrix. The enzymes were furth er separated and purified into 12 components after DEAE-32 chromatography and p reparative electrophoresis. The pI of these components gradually decreased f rom pH 4.0 according to electrophoresis mobility from higher to lower on PAGE. The molecular weights were in the range of 22～34 ku. 6.5 and 7 w ere glycoproteins proved by staining with the shiff reagent and thymol/sulfuric acid. The fibrinolytic activity of 7 was highest as determined usi ng chromzym UK and chromzym PL as specific substrates.