• 한국임상수의학회지
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• 제31권3호
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• pp.170-174
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• 2014

본 연구는 개 소장점막하 조직을 겔화시킨 것과 일반적으로 사용하는 소 트롬빈, 그리고 이 두 가지 요소를 혼합한 것을 혈소판풍부혈장의 활성화 인자로 적용하여, 혈소판에서 분비되는 성장인자를 7일간 in vitro 실험을 실시하였다. 또한 전자현미경으로 각 그룹 별 샘플의 표면을 관찰하여, 측정한 성장인자와의 상관관계를 연구하였다. 그 결과, 그룹별로 유의적인 차이는 존재하지 않았지만, 성장인자인 (TGF)-${\beta}1$이 가장 많이 분비된 그룹은 혈소판풍부혈장을 개 소장점막하조직 겔로 활성화시켰을 때였다. 본 연구 결과를 통해, 혈소판풍부혈장을 활성화하기 위한 요소로 개소장점막하조직 겔은 소의 트롬빈을 대체할 만한 인자로 여겨지며, 추후 임상에서 다양하게 적용할 수 있을 것으로 기대된다.

• 대한의용생체공학회:의공학회지
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• 제16권3호
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• pp.367-375
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• 1995

광범위 항생물질인 Ampicillin sodium(AMP-Na)을 두가지 방법 즉, 단순섞임과 bovine serum albumin(BSA)으로 microsphere화한 후 loading 하는 방법으로 fibrin glue(FG)를 제조하였고 이 FG로부터 AMP-Na의 서방성 시험을 시도하였다. 단순섞임의 경우 fibrinogen(FBNG)의 농도를 조적함으로서 FG로부터 AMP-Na의 지속적 방출을 달성할 수 있었으며 특히 이 microsphere를 glutaraldehyde로 가교화시킴으로서 용출속도를 더욱 늦출 수 있었다.(tO.9 : 33hr). FG의 rat perironeum에 대한 접착력은 FBNG과 thrombin의 농도가 각각 5.0%, 25~50 NIHU/ml에서 최대로 나타났다. Factor XIII의 농도는 0~500 U/1g of FBNG의 범위내에서 접착력에 거의 영향을 미치지 않았으며 incubation time은 60분일 때 최대로 나타났다. FG에 AMP-Na 및 BSA micorsphere를 loading하여도 접착력에는 큰 영향을 미치지 않았다.

• Tuberculosis and Respiratory Diseases
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• 제44권5호
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• pp.1114-1124
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• 1997

연구배경 : Endothelin(이하 ET로 약함)은 폐혈관 내피세포에서 생산되며 강한 혈관 수축작용이 있는 peptide이다. 일차성 폐동맥고혈압과 급성 폐동맥색전증 환자의 혈장내 ET이 증가하고 이들 질환에서의 심폐기능장애에 ET이 중요한 역할을 하리라고 추측되나 ET증가의 기전에 대해서는 알려진 바 없다. 이들 두 질환은 모두 혈전증이 중요한 병태생리 소견이므로 저자들은 혈소판과 활성화된 혈소판에서 유리된 매개체들이 폐혈관 내피세포에서 ET 생산을 증가시킬 것이라고 가정하고 이를 확인하기 위하여 연구를 시행하였다. 방 법 : 소 폐동맥내피세포 배양배지에 혈소판, thrombin(0.1~10u/ml), transforming growth factor-${\beta}1$(TGF-${\beta}1$, 1-1000pM), serotonin(1-100uM), 및 내독소(1ug/ml)를 첨가한 후 18시간 배양하여 배지내로 유리된 immunoreactive ET(이하 ir-ET로 약함)을 방사선면역측정법으로 정량분석 하였다. 결과 : 소 폐동맥내피세포 배양 상청액내 ir-ET은 배양 시간에 비례하여 증가하였으며 혈소판 $10^8/ml$을 첨가한 군에서는 배양 8시간 및 18시간 후에 대조배지군에 비해 유의하게 높았다(p<0.05). 혈소판 첨가군에서 배양 상청액내 ir-ET은 혈소판의 수가 증가함에 따라 증가하는 경향을 보였고 $10^8/ml$에서는 대조배지군에 비해 유의하게 높았다 (0<0.05). Ir-ET를 증가시키지 않는 혈소판 ($10^7/ml$)에 ir-ET을 유의하게 증가시키지 않는 농도의 thrombin (0.1u/ml) 또는 내독소(1ug/ml)를 각각 첨가한 군에서 배양 상청액내 ir-ET은 배지 대조군과 단독 첨가군 (thrombin 0.1u/ml, 내독소 1ug/ml)에 비해 각각 유의하게 높았다(p<0.05). 소 폐동맥내피세포 배양 상청액내 ir-ET은 thrombin(1-10ug/ml), TGF-${\beta}1$(100-1000pM) 첨가군에서 각각 대조배지군에 비해 유의하게 높았으며 (p<0.05), serotonin(1-100uM) 첨가군은 대조배지군파 유의한 차이가 없었다. 결 론 : 소 폐동맥 내피세포에서 ET 생산을 자극하며 그 기전은 혈소판과 활성화된 혈소판에서 유리되는 TGF-${\beta}1$ 등의 매개물에 의한 내피세포의 자극으로 생각되며 이외에 혈소판에 의한 내피세포의 ET 생산 반응 조절(priming) 가능성도 추측할 수 있다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권1호
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• pp.44-47
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• 2000

Cell growth and DNA synthesis were studied from a cultured early- and late- pas- sage mouse aorta smooth muscle cell (MASMC) because the proliferation of vascular smooth muscle cell (VSMC) is a key factor in development of atherosclerosis. In this study, the cells were cultured in fetal bovine serum (FBS) and stimulated by growth factors such as thrombin and platelet-derived growth factor-BB (PDGF-BB). Compared to the number of early-passage MASMC (passage 3 to 9) the number of late-passage MASMC (passage 30 to 40) in a normal serum state was increased 2 fold at Day 1, 3 and 6 in culture, respectively. Incorporation of $[^3H]$ thymidine into DNA induced by serum, PDGF and thrombin in late-passage MASMC was greater than those in early-passage MASMC. We also examined whether intracellular zinc levels would be an aging factor or not. The intracellular zinc level in early- and late-passage MASMC was monitored by using the zinc probe dye N-(6-methoxy-8-quinolyl)-p-toluenesulfonamide. It is interested that late-passage MASMC increased the intracellular fluorescence level of zinc, more than the early passage MASMC did. The alterations of intracellular zinc level occur concurrently with changes in MASMC proliferation rate during aging. This data suggest that the age-associated changes in zinc concentrations may provide a new in vitro model for the study of smooth muscle cell differentiation.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제21권1호
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• pp.81-88
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• 1999

A fibrin adhesive have been widely used in oral and maxillofacial surgery for microvascular anastomosis, autogenous chip bone grafts, many kinds of soft tissue surgery (vestibuloplasty, bleeding control after extraction, primary healing by covering of suture of a gum after the extirpation of large cysts). There are two principal components in adhesive systems biologically: lyophilized human fibrinogen and bovine thrombin. The fibrinogen component contains coagulation factor XIII and enhance the initial wound healing, which polymerizes soluble fibrin monomers into an insoluble clot. The thrombin is dissolved in a solution of calcium chloride to provide the second component. We applied fibrin adhesive, Beriplast (Behring, Behringwerke AG, D-3350, Marburg, FRD), to 4 patients for fixation of free skin grafting donors who had facial scar around eye, nose, mouth corner which received from accidents, or burn. We have experienced initial accelerated graft fixation between donor and recipient sites with no additional fixation. And It's made easy bleeding control and easy manipulation during operation. But two cases showed partial hypertrophic scar engrowth in above 3 months follow up, but no significant. Histopathological reviews in general were showed similar scar findings such as abundant collagen bundles in H&E, M/T stain, but slight positive signs in elastic and collagen antibody immunopathologic findings in hypertrophic scar cases.

• Asian-Australasian Journal of Animal Sciences
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• 제17권8호
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• pp.1062-1068
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• 2004

Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제13권2호
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• pp.129-136
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• 1991

Fibrin Sealants 는 과거 간, 비장, 신장등의 일반외과 수술에서 단순한 외과적 봉합으로는 해결하지 못하는 넓은 면적의 조직의 유착을 위하여 1940 년경 부터 개발되기 시작한 것으로 악안면 영역에서는 1970 년대 중반부터 미세신경접합술과 피부이식을 위하여 사용되기 시작한 후, 현재, 골이식후의 골세편의 고정, 혈관봉합술, 연조직에서 조직들의 유착과 지혈, 그리고 혈관종의 치유등을 목적으로도 광범위하게 연구되고 사용되고 있다. 이것은 인체에서 채취한 혈액응고인자 XIII 을 포함하는 Fibrinogen 성분과, 소에서 추출한 Thrombin 의 두가지 주요 성분으로 구성되며, Fibrinogen 용해제인 Aprotinin 액과 Thrombin 용해제인 염화칼슘액과 함께 네부분으로 구성된다. 각제품에 따라 그리고 사용된 농도에 따라 차이는 있으나, 대개 수분후에 조직이 응고되어 달라붙기 시작하고, 수시간후에 최대접착효과에 도달하며, 응고된 접착효과는 12 일에서 15 일간 유지되고 그후 정상적인 섬유소분해작용과 식세포활동에 의하여 분해된다. 저자는최근 6 개월간 서울대학교병원 구강악안면외과에서 28 명의 각종 질환 및 기형 환자에서 미세신경봉합술, 피부이식, 악교정성형술과 구개파열 또는 하악골 재건을 요하는 환자의 골이식후의 골세편의 고정, 경부곽청술이나 종양제거술후 각종 피부판 또는 근피판을 이용한 연조직의 적합, 혈관종의 처치, 후이개접근 법에 의한 악관절수술후의 외이도의 접합등 다양한 목적을 위하여 적용된 Fibrin Sealants 를 사용하여 양호한 결과를 얻었기에 보고하는 바이다.

• Archives of Plastic Surgery
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• 제39권6호
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• pp.585-592
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• 2012

During the past decade, many studies using platelet-rich plasma (PRP) or adipose-derived stem cells (ASCs) have been conducted in various medical fields, from cardiovascular research to applications for corneal diseases. Nonetheless, there are several limitations of practical applications of PRP and ASCs. Most reports of PRP are anecdotal and few include controls to determine the specific role of PRP. There is little consensus regarding PRP production and characterization. Some have reported the development of an antibody to bovine thrombin, which was the initiator of platelet activation. In the case of ASCs, good manufacturing practices are needed for the production of clinical-grade human stem cells, and in vitro expansion of ASCs requires approval of the Korea Food and Drug Administration, such that considerable expense and time are required. Additionally, some have reported that ASCs could have a potential risk of transformation to malignant cells. Therefore, the authors tried to investigate the latest research on the efficacy and safety of PRP and ASCs and report on the current state and regulation of these stem cell-based therapies.

• 대한수의학회지
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• 제45권2호
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• pp.273-278
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• 2005

This study was investigated of the bone healing capacity by autologous fibrin glue in experimental bone defect dogs. The autologous fibrin glue manufactured just before the experiment was mixed with the concentrated fibrinogen from whole blood of the experimental dog and bovine thrombin. The experimental group was constituted with seven dogs. The experimental osteotomy was performed 5 mm length in bilateral region of proximal diaphyseal fibulae. The defected regions of experimental group were filled with the autologous fibrin glue by duploject. The experimental regions had been radiographed biweekly for 16 weeks to observe new bone formation and union. Bone alkaline phophatase (BALP) in all groups was evaluated biweekly till the end of the experiment to determine osteoblast activities. New bone formation had been observed in five regions of three dogs at four weeks after the experimental treatment and in two regions of one dog at ten weeks. The other seven regions of the experimental group and control group were not observed new bone formation until the end of the experiment. BALP value in four dogs observed new bone formation was increased to 97.10 IU/L (453.96%) at two weeks after the experimental treatment. The results of this experiment were suggested that the autologous fibrin glue was moderately effective in new bone formation in dogs.

• 한국미생물·생명공학회지
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• 제51권2호
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• pp.167-173
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• 2023

Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.