• YAKHAK HOEJI
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• v.45 no.5
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• pp.476-483
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• 2001

The clotting assay was replaced by the chromogenic substrate assay which is recommended by the European Pharmacopoeia (EP) and the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis based on the reliability convenience and simplicity of the chromogenic assay, A correlation study was carried out with a one-stage factor Ⅷ : C clotting assay and the performance of the chromogenic assay was evaluated using two test kits that fulfilled the requirements of EP for factor Ⅷ concentrates test. Although chromogenic assay has partly differences in measurement principle and standardization, this assay has a high correlation with clotting assay in various types of factor Ⅷ concentrates and factor Ⅷ standard. We conclude that the chromogenic assay for factor Ⅷ : C concentrates correlates well with the clotting assay and shows good analytical performance.

• 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.

• Biomolecules & Therapeutics
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• v.1 no.2
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• pp.166-170
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• 1993

Hirudin is a potent inhibitor of thrombin, which was originally obtained from the medicinal leech (Hirudo medicinalis) Now it is being produced through the recombinant technology on a large scale. Recombinant hirudin has been assayed for the anticoagulant activity by the measurement of clotting time and the inhibition of thrombin actvity using a chromogenic substrate. The assay range of partial thromboplastin time and thrombin time is within $0.2{\sim}1.0 {\mu}g/mι.$ Thrombin time is more sensitive to the measurement of clot. Ex vivo study showed the level of hirudin in rat plasma was highest in 10 min and then it was eliminated slowly. The half-life of r-hirudin was 80~110 min depending on the assay methods. Intraveneous injection of russel viper venom was used for thrombus induction combined with vents cava ligation. Inhibition of venous thrombosis was observed with i.v. hirudin. It was dependent on the concentration of hirudin.

• Applied Biological Chemistry
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• v.47 no.1
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• pp.149-153
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• 2004

전통된장 제조방법으로서 원료 혼합비, 메주의 건조시간 및 온도, 메주와 된장의 발효 시간 및 온도, 된장 간장의 분리 여부, 된장 숙성(aging) 시간을 조사하였다. 혈액응고 저해 활성(anticoagulant activity)을 fibrin clotting assay법으로 분석하여 혈액응고 저해활성과 제법간의 상관 관계를 검토하였다. 숙성 기간이 길수록 혈액응고 저해활성이 높게 나타나는 경향을 보였으나 선형적인 상관관계보다는 비선형적으로 양의 상관관계를 갖는 것으로 판단되었다. 이상의 결과를 토대로 혈액응고 저해활성이 높은 2종의 된장시료를 선발하였으며 숙성기간이 180일 이상일 경우 전통된장의 혈액응고 저해활성이 높은 것으로 판단되었다.

• Archives of Pharmacal Research
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• v.24 no.2
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• pp.109-113
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• 2001

Curdlan is a natural $\beta$-1,3-glucan produced by Agrobacterium biovar 1. In this study, the anticoagulant activity of sulfoalkyl derivatives of curdlan was investigated by carrying out activated partial thromboplastin time (APTT) assay and compared with that of o-sulfonated curdlan. Approximately 100-fold higher concentration of o-sulfonated curdlan than heparin was required to obtain the same level of the clotting time. Anticoagulant activity of curdlan derivatives was dependent on the degree of sulfation in prolonging the clotting time. However, the chain length of the substituent did not play a role in prolonging the clotting time. The curdlan derivatives enhanced thrombin inhibition by mediating through antithrombin III. The inhibition of thrombin by o-sulfonated curdlan was found to be approximately 10-fold weaker than that by heparin.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.139-147
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• 2011

Objectives : The aim of this study is to research an anti-thrombus effect by Diospyros Kaki Calyx. Methods : The healthy human plasma were gained and used 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 (orally administrated with distilled water 5ml/kg) and two experimental group treated with extract of diospyros kaki calyx (EKC). Experimental rats were orally 600 mg/kg concentration of EKC and 200 mg/kg concentration of EKC. After an hour from administration, we anesthetized rats and made arteriovenous (AV) shunt rat models to study weight of thrombus, took whole blood to study content of thromboxane B2 and blood clotting time. Results : In vitro, EKC significantly increased inhibitory activity of FXa, prothrombinase compared with intact control group ($^*P$ <0.05). PT and aPTT were increased in EKC treated (600 mg/kg) group compared with intact control group ($^*P$ <0.05). In vivo, blood clotting time of experiment group treated with EKC 600 mg/kg were significantly increased compare with that of intact control group (p<0.05) and content of thromboxane B2 was significantly decreased in group treated with EKC 600 mg/kg in serum. The weight of thrombus were significantly reduced in group treated with EKC 600 mg/kg compared with intact control group (p<0.05). But in vivo experiment study, those parameters of group treated with EKC 200 mg/kg were relatively decreased compared with those of intact control group without statistical significance. Conclusions : EKC has an antithrombic activity because of inhibition internal course such as FXa and prothrombin. And EKC inhibited a hole blood clotting in vivo experiment by low content of thromboxane B2.

• Korean Journal of Pharmacognosy
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• v.36 no.4 s.143
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• pp.305-310
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• 2005

Sarcodon aspratus (Thelepholaceae), a native mushroom, is distributed in Korea and Japan, and has been widely used in traditional food and fork medicines. To confirm the biological activities of Sarcodon aspratus, the liver protecting activity, anti-clotting activity, and anti-complementary activity of the water extract, EtOH extract, and the water soluble proteoglycan part of S. aspratus were investigated. The EtOH, and water extract of S. aspratus decreased the GOT and GPT releases induced by $CCl_4$ in a dose-dependant manner. On the other hand, the water soluble proteoglycan part of S. aspratus showed weak inhibitory activity. In addition, the EtOH and water extract of S. aspratus prevented $CCl_4-induced$ hepatotoxicity, as described by a liver histopathologic study. To confirm the anti-clotting activity, the APTT and PT assay were carried out. As a result, only the crude proteoglycan part of S. aspratus showed the anti-coagulating activity, and this result might be due to the inhibition of intrinsic clotting system. Also, the crude proteoglycan part of S. aspratus showed the anti-complementary activity, and the $IC_{50}$ value was $50\;{\mu}l/ml$.

• Archives of Pharmacal Research
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• v.27 no.6
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• pp.619-623
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• 2004

Tissue factor (TF, tissue thromboplastin) is a membrane bound glycoprotein, which acceler-ates the blood clotting, activating both the intrinsic and the extrinsic pathways to serve as a cofactor for activated factor VII (Vila). The TF-factor Vila complex (TF/VIIa) proteolytically activates factors IX and X, which leads to the generation of thrombin and fibrin clots. In order to isolate TF inhibitors, by means of a bioassay-directed chromatographic separation technique, from the leaves of Eriobotrya japonica Lindley (Rosaceae), a known sesquiterpene glycoside (2) and ferulic acid (3) were isolated as inhibitors that were evaluated using a single-clotting assay method for determining TF activity. Another sesquiterpene glycoside (1) was also isolated but was inactive in the assay system. Compound 3 was yielded by alkaline hydrolysis of compound 2. The structures of compounds 1, 2, and 3 were identified by means of spectral analysis as $3-O-{\alph}-L-rhamnopyranosyl-(1{\rightarrow}4)-a-L-rhamnopyranosyl-(1{\rightarrow}2)-[{\alph}-L-rhamnopyrano-syl-(1{\rightarrow}6)]-{\beta}-D-glucopyranosyl nerolidol$ (1), $3-O-{\alph}-L-rhamnopyranosyl-(1{\rightarrow}4)-{\alph}-L-rhamnopyr-anosyl-(1{\rightarrow}2)-[{\alph}-L-(4-trans-feruloyl)-rhamnopyranosyl-(1{\rightarrow}6)]-{\beta}-D-glucopyranosyl$ nerolidol (2) and ferulic acid (3), respectively. Compounds 2 and 3 inhibited 50% of the TF activity at con-centrations of 2 and $369{\;}\mu\textrm{m}/TF$ units, respectively.

• Restorative Dentistry and Endodontics
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• v.47 no.4
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• pp.42.1-42.11
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• 2022

Objective: This study investigated the effects of various concentrations of sodium hypochlorite (NaOCl) on human whole-blood clotting kinetics, the structure of the blood clots formed, and transforming growth factor (TGF)-β1 release. Materials and Methods: Human whole blood was collected from 5 healthy volunteers and divided into 4 groups: CG (control, 0.5 mL of blood), BN_0.5 (0.5 mL of blood with 0.5 mL of 0.5% NaOCl), BN₃ (0.5 mL of blood with 0.5 mL of 3% NaOCl), and BN_5.25 (0.5 mL of blood with 0.5 mL of 5.25% NaOCl). The effects of NaOCl on clotting kinetics, structure of fibrin and cells, and release of TGF-β1 were assessed using thromboelastography (TEG), scanning electron microscopy (SEM), and enzyme-linked immunosobent assay, respectively. Statistical analysis was conducted using the Kruskal Wallis and Mann-Whitney U tests, followed by the post hoc Dunn test. A p value < 0.05 indicated statistical significance. Results: The blood samples in BN_0.5 and BN₃ did not clot, whereas the TEG of BN_5.25 showed altered clot formation. Samples from the CG and BN₃ groups could only be processed with SEM, which showed that the latter lacked fibrin formation and branching of fibers, as well as clumping of red blood cells with surface roughening and distortion. TGF-β1 release was significantly highest in BN₃ when all groups were compared to CG (p < 0.05). Conclusions: Each concentration of NaOCl affected the release of TGF-β1 from blood clots and altered the clotting mechanism of blood by affecting clotting kinetics and cell structure.

• Journal of Nutrition and Health
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• v.28 no.9
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• pp.862-871
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• 1995

This study was designed to investigate the effectiveness of ginseng intakes in modifying serum lipid profiles and plasma clotting factors. The participants in this study were 47 normal healthy volunteers(men 24, women 23) with an age range of 35-49 years and a mean age of 41 years residing in Taejon. Based on the diet intakes, subjects were classed into one of three groups : control, vegetarian, and ginseng consumed over 3-4 years. There was no significant difference in their physical characteristics. Dietary calorie intakes were not significantly different in subjects. The ratio of energy intake in the control and ginseng consumed group was 63-64% : 20-21% : 15-16%(Cho : Fat : Pro), but 70-73% : 13-14% :14-15%(Cho : Fat : Pro) in the vegetarians. The intakes of animal food in the vegetarian was significantly lower than the control and ginseng consumed group in men. The ratio of P/S(1.27) was the highest in the vegetarians. Venous blood samples were taken for serum lipid profiling, plasma clotting assay and platelet function. The concentration of serum triglyceride in the men ginseng group is significantly lower than those of the men control group. Serum lipid profiles values of the men ginseng group, such as total cholesterol and phospholipid were lower those of the men control group, but higher those of the men vegetarian group. the serum lipid profile in the women were not significant, but total cholesterol, triglyceride and LDL cholesterol levels in the ginseng groups were low. The concentration of HDL cholesterol was not significantly different. Platelet cell count and platelet aggregation were low in the ginseng groups. APTT(Activated Partial thromboplastin time) was significantly elongated in ginseng groups in the normal range. In seems that the major beneficial effects of ginseng intakes in especially men were on the blood concentrations of triglyceride, total cholesterol and elongation of plasma clotting time.