• Title/Summary/Keyword: Clotting factor

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Potency Assay of Factor Ⅷ : C Concentrates using the Chromogenic and Clotting Assay (합성기질 및 응고시간을 이용한 혈액응고 제 8인자 역가측정법)

  • 강혜나;김순남;허숙진;홍성화
    • 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.

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A Case Report of Palatoplasty in a Patient with Clotting Factor Ⅹ Deficiency (선천성 혈액 응고 인자 Ⅹ(10번) 결핍증 환아에서 구개성형술의 증례보고)

  • Kim, Suk Wha;Jeong, Eui Cheol;Yun, Byung Min;Choi, Tae Hyun;Kang, Hyoung Jin
    • Archives of Plastic Surgery
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    • v.36 no.6
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    • pp.792-794
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    • 2009
  • Purpose: Clotting factor X deficiency is one of the most uncommon coagulation disorders. The authors describe a case of cleft palate in a patient with a congenital clotting factor X deficiency. Methods: In pediatric patients with a cleft palate, the coagulation problem is more worrisome, because they are more sensitive to blood than adults, and because postoperative bleeding can cause blood ingestion with subsequent vomiting, aspiration, and airway obstruction. To prevent hemorrhagic complications in the described case, fresh frozen plasma (FFP) was administered every 24 hrs from the day before surgery to the second postoperative day. Results: Good hemostasis, normal healing, and no complications was shown postoperatively. Conclusion: The replacement of fresh frozen plasma was useful in the case of congenital clotting factor deficiency for bleeding prophylaxis in cleft palate operation.

Anticoagulant Activity of Ilexoside D, a Triterpenoid Saponin from ilex pubescens

  • Han, Yong-Nam;Song, Jae-Ihn;Rhee, In-Kyung
    • Archives of Pharmacal Research
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    • v.16 no.3
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    • pp.209-212
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    • 1993
  • The anti-coagulant activity of ilexoside D isolated from the roots of ilex pubescens Hook. et Am. was investigated in in vivo models of blood coagulation in rats. On oral administration, ilexoside D prolonged the bleeding time and the whole blood recalcified clotting time, but not the plasma recalcified clotting time. In vitor, ilexoside D did not affect the recalciffed clotting times of whole blood, platelet-rich plasma(PRP), and platelet-poor plasma(PPP), while in the presence of tissue factor the compound prologed the reduced proth-rombin times of whole blood, PRP and PPP in the dose-dependent manner. These results indicate that ilexoside D has the anit-tissue factor activity as well as the antithromobotic activity.

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Effects of Curculiginis Rhizoma on anti-thrombotic activity (선모(仙茅) 열수(熱水) 추출물(抽出物)의 항혈전(抗血栓) 효능 연구)

  • Roh, Seong-Soo
    • 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.

Honey Bee Venom (Apis mellifera) Contains Anticoagulation Factors and Increases the Blood-clotting Time

  • Zolfagharian, Hossein;Mohajeri, Mohammad;Babaie, Mahdi
    • Journal of Pharmacopuncture
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    • v.18 no.4
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    • pp.7-11
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    • 2015
  • Objectives: Bee venom (BV) is a complex mixture of proteins and contains proteins such as phospholipase and melittin, which have an effect on blood clotting and blood clots. The mechanism of action of honey bee venom (HBV, Apis mellifera) on human plasma proteins and its anti-thrombotic effect were studied. The purpose of this study was to investigate the anti-coagulation effect of BV and its effects on blood coagulation and purification. Methods: Crude venom obtained from Apis mellifera was selected. The anti-coagulation factor of the crude venom from this species was purified by using gel filtration chromatography (sephadex G-50), and the molecular weights of the anti-coagulants in this venom estimated by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Blood samples were obtained from 10 rabbits, and the prothrombin time (PT) and the partial thromboplastin time (PTT) tests were conducted. The approximate lethal dose (LD) values of BV were determined. Results: Crude BV increased the blood clotting time. For BV concentrations from 1 to 4 mg/mL, clotting was not observed even at more than 300 seconds, standard deviations $(SDs)={\pm}0.71$; however, clotting was observed in the control group 13.8 s, $SDs={\pm}0.52$. Thus, BV can be considered as containing anti-coagulation factors. Crude BV is composed 4 protein bands with molecular weights of 3, 15, 20 and 41 kilodalton (kDa), respectively. The $LD_{50}$ of the crude BV was found to be $177.8{\mu}g/mouse$. Conclusion: BV contains anti-coagulation factors. The fraction extracted from the Iranian bees contains proteins that are similar to anti-coagulation proteins, such as phospholipase $A_2(PLA_2)$ and melittin, and that can increase the blood clotting times in vitro.

The influence of sodium hypochlorite concentration on the fibrin structure of human blood clots and transforming growth factor-beta 1 release: an ex vivo study

  • Anisha Mishra ;Velmurugan Natanasabapathy;Nandini Suresh
    • 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), BN0.5 (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 BN5.25 (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 BN0.5 and BN3 did not clot, whereas the TEG of BN5.25 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.

Dental Treatment of Child with Hemophilia (혈우병을 가진 어린이의 치과치료)

  • Lim, Ji Eun;Lee, Soo Eon;Ahn, Hyo Jung;Park, Jae-Hong;Choi, Sung Chul
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.12 no.4
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    • pp.229-233
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    • 2012
  • Hemophilia, the most common of the inherited bleeding disorder, is the result of a deficiency of clotting factor. Since bleeding after dental treatment may cause severe or even fatal complications, people with hemophilia must be given special dental care. We report on the diagnosis and treatment of a 9-year-old boy having severe hemophilia visited our department with the chief complaints of pus discharge on the left lower molar region. In the clinical and radiographic examination, periapical abscess and dental caries were diagnosed. Considering complexity of the treatment and complication in the coagulation, it was decided to carry on the treatment under general anesthesia. Clotting factor IX concentrates were intended to provide 50-70% plasma level. Pulpectomy, resin restoration and Stainless steel crown were given under general anesthesia. Several teeth were extracted and the sockets were packed with Surgicel$^{(R)}$ (Oxidized Regenerated Cellulose, Johnson and Johnson Co. Neuchatel, Switzerland) under general anesthesia. Transpalatal arch and lingual arch were given for maintaining the extracted space before discharged. For people with severe hemophilia, factor replacement is necessary before scaling, surgery or regional block injections. Therefore, if several extractions are needed, dental care under general anesthesia would be effective and efficient management.

Tissue Factor Inhibitory Sesquiterpene Glycoside from Eriobotrya japonica

  • Lee, Ming-Hong;Son, Yeon-Kyoung;Han, Yong-Nam
    • 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.

Cloning and Expression of Human Clotting Factor 9 cDNA un Escherichia coli (인체 혈액응고 9인자 cDNA cloning 및 Escherichia coli 에서의 발현)

  • Young Won Lee;Hyang Suk Hur;Myoung Hee Kim
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.231-240
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    • 1996
  • Human blood clotting (coagulation) factor 9 cDNA which codes for 461 amino acid has been cloned by screening human fetal liver cDNA library using PCR. This 1.4 kb cDNA spanning from the ATG initiation codon to the TAA termination codon was cloned into bacterial .expression vector pGEX-2T, generating pGEX-F9 plasmid. The plasmid pGEX-F9 expresses about 73 kDa GST (Glutathione S-transferase)-Factor 9 fusion protein when introduced into E. coli. Western blot analysis using polyclonal antibody raised against human factor 9 confirmed this fusion protein contains factor 9 protein. The level of GST-factor 9 expression was about 20% of total protein and the purification of fusion protein was efficiently achieved by using GST agarose bead based on one step purification protocol.

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Effects of Aqueous Extract of Diospyros Kaki Calyx on Anti-thrombotic Activity in vitro and in vivo (시체(柿蒂)의 in vitro와 in vivo 항혈전 효능 연구)

  • Baek, Kyung-Min;Roh, Seong-Soo
    • 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.