• Journal of Life Science
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• v.17 no.4 s.84
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• pp.535-539
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• 2007

The fructus of Crataegus pinnatifida Bunge has been used as medicinal and food source in worldwide. In this study, a strong direct thrombin inhibition and antithrombosis activity were identified from the methanol extract of C. pinnatifida Bunge fructus. The solvent fractionation of fructus extract using hexane, ethylacetate, butanol revealed that the butanol fraction has a prominent antithrombin activity. Thrombin time(blood-clot formation time) and activated partial thromboplastin time(aPTT) extended to 835% and 315% by addition of the butanol fraction at concentration of 1.25 mg/mL, whereas thrombin time extended to 287% by addition of aspirin at concentration of 1,25 mg/mL. The butanol fraction showed anthrone-positive and weak ninhydrine-postive reaction. The thrombin inhibitory activity was not related to previously reported flavonoids or polyphenols. The activity was maintained against acid treatment(0.5 N HCl for 120 min), but rapidly lost by heat-treatment($100^{\circ}C$ for 30 min). Our results suggested that fructus of C. pinnatifida Bunge with non-heat treatment process could be developed as a natural source of antithrombosis.

• Journal of Life Science
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• v.20 no.6
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• pp.922-928
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• 2010

This study was carried out to develop safe and novel anticoagulation agents from oriental medicinal herbs. From 33 medicinal herbs, 40 different ethanol extracts were prepared according to place of origin or extraction parts, and anticoagulation activities were evaluated by determination of thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (aPTT). The average water content and average extraction ratio for the medicinal herbs were $6.85{\pm}2.26%$ and $5.27{\pm}4.25%$, respectively. Evaluation of TT at various concentrations of the extract led to the selection of Mucuna birdwoodiana, Prunus armeniaca, Cacalia ainsliaeiflora, Cinnamonum aromaticum, and Rhus javanica Linneas potent antithrombosis medicinal herbs. Evaluation of PT and aPTT showed that the extracts of R.javanica Linne, M. birdwoodiana, and P. armeniaca have strong anticoagulation activities. Determination of hemolytic activities of 40 different ethanol extracts against human red blood cells, however, showed that only M. birdwoodiana, C. ainsliaeiflora, C. aromaticum, and R. javanica Linnehas strong anticoagulation activity without hemolytic activity at a concentration of 500 mg/ml. Our results suggest that oriental medicinal herbs, which are under a mass-production system, have potentialas a safe and novel source of anticoagulants, as well being a thrombin-specific and coagulation factor-specific inhibitor.

• Journal of Life Science
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• v.20 no.11
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• pp.1640-1647
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• 2010

Seaweeds have been recognized as a health food, having anti-obesity, anti-constipation and anticoagulation activities, and the use of seaweeds in the food, medicine, and cosmetic industries have recently significantly increased. In this study, methanol extracts were prepared from 35 different seaweeds (17 phaeophyta, 11 rhodophyta and 7 chlorophyta), and thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (aPTT) were determined in order to develop safe and novel anticoagulation agents from natural products. In TT experiments, Ecklonia cava, Ecklonia stolonifera, Eisenia bicyclis (Kjellman) Setchell, Ishige foliacea, I. okamurai, Sargassum confusum and S. yamade showed strong thrombin inhibition activity among the 35 different seaweeds. In PT experiments, the inhibitions of prothrombin were identified in the selected seaweeds from TT experiment, with the exception of S. yamade. In aPTT experiments, the seaweeds with blood coagulation inhibition factors were E. cava, E. stolonifera, E. bicyclis (Kjellman) Setchell, I. foliacea, I. okamurai, S. confusum and Hixikia fusiforme Okamura. Further anticoagulation assay with the selected 8 seaweeds suggested that S. confusum is most effective in antithrombosis, and E. stolonifera, E. bicyclis (Kjellman) Setchell, and I. foliacea have high potential as antithrombosis agents. Based on components-activity correlation analysis, flavonoids are considered as active anticoagulation components of seaweeds These results suggest that edible seaweeds, especially S. confusum, have potential as safe and novel anticoagulants, and S. yamade and H. fusiforme Okamura could be used as a thrombin-specific and coagulation factor-specific inhibitors.

• Journal of Haehwa Medicine
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• v.16 no.2
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• pp.303-310
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• 2007

The purpose of this study is to investigate the anti-coagulant effects of Polygoni Mutiflori Radix extracts. The results were as follows : 1. This study show that Polygoni Mutiflori Radix extracts have Anti-coagulant significant effects on Prothrombin time in vitro test. 2. This study show that Polygoni Mutiflori Radix extracts have Anti-coagulant significant effects on Activated Partial Thromboplastin Time vitro test.

• Journal of Korean Neurosurgical Society
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• v.42 no.5
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• pp.377-381
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• 2007

Objective : High-molecular-weight hydroxyethyl starch (HES) compromises blood coagulation more than does low-molecular-weight HES. We compared the effects of low- and high-molecular-weight HES for the treatment of vasospasm and investigated the dose relationship with each other. Methods : Retrospectively, in a series of consecutive 102 patients with subarachnoid hemorrhage (SAH), 35 patients developed clinical symptoms of vasospasm of these fourteen patients were treated with low-molecular weight HES for volume expansion while the other 21 received high-molecular-weight HES as continuous intravenous infusion. Prothrombin time (PT), partial thromboplastin time (PIT), fibrinogen level, and platelet count were all measured prior to initiation, during treatment and after termination of therapy for symptomatic vasospasm. The total dose of HES ranged from 5 L to 14 L and median infusion duration was 10 days. Results : A more pronounced PTT prolongation was observed in high-molecular-weight HES group compared with low-molecular-weight HES group. No other coagulation parameters were altered. Dosage (=duration) shows a positive correlation with PTT. Clinically, significant bleeding episodes were noted in four patients who received high-molecular-weight HES. Conclusion : Coagulopathy was developed in direct proportion to molecular weight of starch and dosages. We propose the extreme caution in the administration of HES solution for the vasospasm treatment.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.2
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• pp.168-173
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• 2012

We performed this study to develop antithrombotic agents from oriental medicine herb extracts. Polygonum cuspidatum has been traditionally used as an edible medical resources for the treatment of cancer, pyodermatitis, hepatitis, cystitis, and inflammation. However, the effects of Polygonum cuspidatum on thrombosis and platelet activation are not precisely understood. The antithrombotic and antiplatelet activities of Polygonum cuspidatum were investigated by assessing the effect of a 70% ethanol extract of Polygonum cuspidatum on blood coagulation, fibrinolysis, and platelet aggregation. Polygonum cuspidatum showed effective fibrinolytic activity at 10 mg/mL. Polygonum cuspidatum also inhibited adenosine diphosphate induced platelet aggregation. Furthermore, evaluation of anticoagulant activity showed that an extract of Polygonum cuspidatum prolonged coagulation time via activated partial thromboplastin time (APTT). Our results show that Polygonum cuspidatum can be a potential candidate for antiplatelet activity as well as a fibrinolytic agent.

• Archives of Pharmacal Research
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• v.29 no.5
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• pp.418-423
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• 2006

Tabanus anticoagulant protein (TAP) was isolated from the whole body of the tabanus, Tabanus bivittatus, using three purification steps (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and ion exchange chromatography on DEAE Sephadex gel). The purified TAP, with a molecular weight of 65 kDa, was assessed to be homogeneous by SDS-polyacrylamide gel electrophoresis, and an isoelectric point of 7.9 was determined by isoelectric focusing. The internal amino acid sequence of the purified protein was composed of Ser-Leu-Asn-Asn-Gln-Phe-Ala-Ser-Phe-lle-Asp-Lys-Val-Arg. The protein was activated by $Cu^{2+}\;and\;Zn^{2+}$, and the optimal conditions were found to be at pH $3\sim6\;and\;40\sim70^{\circ}C$. Standard coagulation screen assays were used to determine thrombin time and activated partial thromboplastin time. Chromogenic substrate assays were performed for thrombin and factor Xa activity. TAP considerably prolonged human plasma clotting time, especially activated partial thromboplastin time in a dose-dependent manner; it showed potent and specific antithrombin activity in the chromogenic substrate assay. Specific anti-factor Xa activity in TAP was not detected. Overall, this result suggested that TAP has significant anticoagulant activity on blood coagulation system.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.125-132
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• 2011

Objectives : An aim of study is to investigate effects of curculiginis rhizoma in vitro (factor Xa (FXa) inhibitor assay, prothrombinase assay, prothrombin time (PT) assay, activated partial thromboplastin time (aPTT) assay) and in vivo experiment (blood clotting time, thromboxane B2 content assay in serum and weight of thrombus by AV-shunt rat model). Methods : We gained a human serum and used serum in vitro study such as factor X activity (FXa) inhibition, prothrombinase inhibition, prothrombin time (PT) and activated partial thromboplastin time. Fifteen SD rats were divided into three groups (intact control group and two experimental group treated with extract of Curculiginis Rhizoma(ECR)). Rats were orally administrated DW (intact control group), 600 mg/kg concertration of ECR and 200 mg/kg concertration of ECR. After one hour, we anesthetized rats and made arteriovenous (AV) shunt rat models to study weights of thrombus, took a hole blood to study content of thromboxane B2 and blood clotting time. Results : In vitro, ECR increased a inhibitory activity of FXa, prothrombinase and aPTT compared than intact control group. Especially ECR made significant increase of FXa and prothrombinase inhibitory activity (p<0.05, p<0.01). And PT were increased in ECR control group compared with intact control group. In vivo, a blood clotting time of experiment group treated with ECR 600 mg/kg were significantly increased compared with that of intact control group (p<0.05) and content of thromboxane B2 was significantly decreased in group treated with ECR 600 mg/kg in seum. The weight of thrombus were significantly reduced in group treated with ECR 600 mg/kg compared with intact control group (p<0.05). But in vivo experiment study, those of group treated with ECR 200 mg/kg were reduced compared with those of intact control group without statistical significance. Conclusions : ECR has a antithromboic activity in internal course with inhibitory activity of FXa and prothrombinase in vitro, it required to research more study for effective compounds.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.2
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• pp.335-341
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• 2000

In order to search for blood anticoagulant substance from herbal extracts, we extracted with cold water (Fr. I), methanol (Fr. II) and hot water (Fr. III) from 93 commercially available herbs. The anticoagulant activity of herbal extracts was examined through the intrinsic pathway by activated partial thromboplastin time (APTT), the extriansic pathway by prothrombin time (PT) and the common pathway by thrombin time (TT). Onion, garlic, clove, fenugreek, mugwort, thyme and sage had on anticoagulant activity with intrinsic pathway and mint, italian seasoning, rosemary, turmeric, tarragon and wasabie had on anticoagulant activity with common pathway. Nobody had on anticogulant activity with extrinsic pathway. Among commerical herbs tested, clove cultivated in holland was selected because it showed the most potent anticoagulant activity among the samples investigated.

• Food Engineering Progress
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• v.13 no.4
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• pp.326-334
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• 2009

Blood coagulation and aggregation of platelet are crucial events in the pathogenesis of various ischemic diseases. The substance which can prevent blood coagulation and platelet aggregation was extracted from wood ear mushroom (Auricularia auricular-judae) and its anticoagulation activity was investigated. The dried A. auricular-judae was extracted with 0.1 N NaOH and its supernatant was further extracted with methanol and ethanol followed by $H_{2}O$. The resulting methanol soluble fraction showed significant antithrombotic activity in activated partial thromboplastin time, thrombin time, and prothrombin time assays with values of 100, 124, and 54 sec, respectively. This active substance was purified with DEAE-Sepharose CL6B and Sephacryl 400-HR and was found to be polysaccharide with the average molecular weight of over 150 kDa. This polysaccharide was xyloglucomannan of which the main component was mannose, and its anticoagulant activity was mostly mediated by inhibition of thrombin activity.