• 대한임상검사과학회지
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• 제43권4호
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• pp.133-137
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• 2011

The monitoring of heparin therapy is using almost aPTT assay. This study is compare to estimating aPTT therapeutic range using in vitro heparin-spiked sample and aPTT therapeutic range using in vivo heparin-treated sample. Normal pooled plasma was collected from 20 healthy representative individuals. 11 concentration of heparinized plasmas from 0 U/mL to 1.0 U/mL at intervals of 0.1 U/mL made by addition of heparin to normal pooled plasma were measured aPTT. The aPTT therapeutic range was performed through correlation analysis between heparin level 0.2 to 0.4 U/mL and aPTT. 30 plasmas from patients on heparin therapy were measured aPTT and anti-Xa activity. The aPTT therapeutic range was performed through correlation analysis between anti-Xa activity 0.3 to 0.7 U/mL and aPTT. The aPTT therapeutic range corresponded by heparin level-vs-aPTT value regression analysis was 60.7 to 102.4 seconds. The aPTT therapeutic range corresponded by anti-Xa activity-vs-aPTT value regression analysis was 85.3 to 147.5 seconds. The validation of heparin sensitivity using in-vitro heparin sample was not considered. The establishing aPTT therapeutic range is recommended anti-Xa activity using in-vivo sample.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.663-669
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• 2000

The fur (ferric uptake regulation) expression vector pMON2064 was modified to produce a Fur-fusion expression vector. A kinker site, factor Xa cleavage site, and several restriction endonuclease sites were introduced to facilitate easy cloning and isolating of the fusion protein. The resulting fusion expression vector, pMONSTER, was then used to make fusion expression vector, pMONSTER, was then used to make fusion proteins with $\beta$-galactosidase and the protease of the human immunodeficiency virus type 1 (HIV-1 PR). Strain SW4020 harboring the Fur $\beta$-galactosidase fusion vector produced blue colonies on a 5-bromo-4-chloro-3-indolyl-$\beta$-D-galactoside plate and the resulting 133 kDa fusion protein reacted with an anti-Fur antibody. The strain harboring the Fur-HIV-1 PR fusion vector produced a 29 kDa fusion protein, which also reacted with an anti-Fur antibody. The Fur-HIV-1 PR fusion protein was purified by a single column application that was designed to isolate the Fur protein. The purified Fur-HIV-1 PR fusion protein digested with factor Xa cleaved a recombinant Gag protein to release smaller fragments, including a p24 capsid protein. The Fur-HIV-1 PR fusion protein itself did not exhibit any proteolytic activity.

• 대한본초학회지
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• 제26권4호
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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.

• Archives of Pharmacal Research
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• 제29권5호
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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.

• 대한본초학회지
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• 제26권4호
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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.

• 생명과학회지
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• 제29권1호
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• pp.37-44
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• 2019

조릿대 잎은 항염, 해열, 이뇨작용 등의 약리효과를 가지고 있어 예로부터 전통의약에서 사용되어 왔다. 본 연구팀은 열수 추출한 후 남는 잔사로부터 식물화합물을 다량으로 함유한 잔사 추출물(PRE)을 제조하는 방법을 보고 바 있다. 본 연구는PRE의 고요산 혈증 저감소재로서 활용 가능성을 평가하기 위하여 수행하였다. Potassium oxonate(PO)로 유도한 고요산 혈증 생쥐 모델에서 PRE는 혈액 내의 요산, 요소 질소, 크레아틴 농도는 감소시켰고, 오줌 내의 요산과 크레아틴 농도는 증가하였다. 또한, PRE 투여한 고용산 혈증 생쥐에서 간 내 요산 농도와 xanthine oxidase 활성이 대조군에 비해 감소하였고, PRE는 PO에 의해 유도된 간 조직의 상해를 보호하였다. 이 결과는 PO로 유도된 고요산 혈증 생쥐에서 PRE는 항염증 및 세포보호 작용에 기인하는 것으로 판단된다. 부가적으로 PRE에 의한 신장조직에서 transcriptome의 반응 변화를 RNA 서열분석법으로 분석하였다. PRE는 주로 면역반응, 염증반응 및 대사과정에 관여하는 유전자의 발현에 영향을 미치는 것으로 나타났다. 본 연구 결과는 염증을 동반하는 고요산 혈증을 개선하는 소재로서의 PRE의 활용 가능성을 제시해 준다.