• 한국수정란이식학회지
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• 제31권3호
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• pp.161-168
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• 2016

Nucleotide metabolism in endothelium is variable between different species. Recent studies demonstrated that this variability could contribute coagulation dysfunction, even though organs of the alpha 1,3-galactosyltransferase gene knockout pig were transplanted into the primate. CD73 (ecto-5'-nucelotidase) is an enzyme at cell surface catalyzing the hydrolysis of adenosine triphosphate to adenosine, which plays role on a substance for anti-inflammatory and anti-coagulant. Thus, overexpression of CD73 in endothelial cells of the pig is considered as an approach to reduce coagulopathy. In this study, we constructed a human CD73 expression vector under control of porcine Icam2 promoter (pIcam2-hCD73), which is expressed specifically at endothelial cells, and of CMV promoter as a control (CMV-CD73). First, we transfected the CMV-CD73 vector into HEK293 cells, and then confirmed CD73 expression at cell surface by flow cytometry analysis. Next, we transfected the pIcma2-CD73 and CMV-CD73 vectors into primary porcine fibroblasts and endothelial cells. Consequence was that the pIcma2-CD73 vector was expressed only at the porcine endothelial cells, meaning that the pIcam2 promoter lead to endothelial cell-specific expression of CD73 in vitro. Finally, we nucleofected the pIcam2-hCD73 vector into passage 3 fibroblasts, and enforced hygromycin selection of 400mg/ml. We were able to obtain forty three colonies harboring pIcam2-CD73 to provide donor cells for transgenic cloned porcine production.

• 한국수정란이식학회지
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• 제33권3호
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• pp.129-137
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• 2018

Acute vascular rejection has been known as a main barrier occurring in a xenograted tissue of alpha 1,3-galactosyltransferase knock-out (GalT KO) pig into a non-human primate (NHP). Adenosine which is a final metabolite following sequential hydrolysis of nucleotide by ecto-nucleotidases such as CD39 and CD73, act as a regulator of coagulation, and inflammation. Thus xenotransplantation of CD39 and CD73 expressing pig under the GalT KO background could lead to enhanced survival of recipient NHP. We constructed a human CD39 and CD73 expression cassette designed for endothelial cell-specific expression using porcine Icam2 promoter (pIcam2-hCD39/hCD73). We performed isolation of endothelial cells (pAEC) from aorta of 4 week-old GalT KO and membrane cofactor protein expressing pig ($GalT^{-MCP/-MCP}$). We were able to verify that isolated cells were endothelial-like cells using immunofluorescence staining analysis with von Willebrand factor antibody, which is well known as an endothelial maker, and tubal formation assay. To find optimal condition for efficient transfection into pAEC, we performed transfection with GFP expression vector using four programs of nucleofection, M-003, U-023, W-023 and Y-022. We were able find that the program W-023 was optimal for pAEC with regard to viability and transfection efficiency by flow cytometry and fluorescent microscopy analyses. Finally, we were able to obtain $GalT^{-MCP/-MCP}/CD39/CD73$ pAEC expressing CD39 and CD73 at levels of 33.3% and 26.8%, respectively. We suggested that pACE isolated from $GalT^{-MCP/-MCP}$ pig might be provided as a basic resource to understand biochemical and molecular mechanisms of the rejections and as an alternative donor cells to generate $GalT^{-MCP/-MCP}/CD39/CD73$ pig expressing CD39 and CD73 at endothelial cells.

• 한국발생생물학회지:발생과생식
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• 제21권2호
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• pp.157-165
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• 2017

One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-human-primate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were $127{\pm}18.9$. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as ${\alpha}-1,3-galactosyltransferase$ knock-out /hCD46 for xenotransplantation.

• 대한소아치과학회지
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• 제46권3호
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• pp.337-342
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• 2019

이 연구의 목적은 발치한 매복 과잉치에서 얻은 치수 유래 세포의 줄기세포 특성을 유세포 분석을 통해 알아보는 것이다. 15명의 환자로부터 채취한 정중 과잉치의 치수 세포를 계대 배양하였고, 3계대와 10계대 세포들을 유세포 분석을 이용하여 분석하였다. 간엽성 줄기세포 표지자 관찰에 사용된 항체 표지자(CD)는 양성 표지자로는 CD 73, CD 90, 그리고STRO-1 와 음성 표지자로는 CD 34, CD 45 이었다. 3계대에서 CD 73, CD 90은 각각 94.82%, 98.86%의 양성반응을, CD 34, CD 45는 각각 2.25%, 2.52%로 음성 반응을 보였으며, STRO-1은 20.93%를 나타냈다. 10계대에서는 CD 73, CD 90은 각각 96.62%, 98.61%의 발현을 보였지만, CD 34, CD 45는 각각 3.86%, 4.14%를 나타냈다. STRO-1은 35.62%로 발현되었다. 이상의 결과에서 과잉치 치수 유래 세포는 간엽성 줄기세포의 특성을 가지며, 3계대와 10계대 모두에서 간엽성 줄기세포의 특성을 유지하고 있다고 사료된다. 이에 빠른 성장 속도와 늦은 계대까지 유지되는 줄기세포능을 고려할 때, 치아 유래 줄기세포의 공여부로서 매복 과잉치의 충분한 활용 가능성을 확인하였다.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2001년도 추계 학술발표강연 및 논문개요집
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• pp.73-73
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• 2001

• 대한약침학회지
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• 제23권4호
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• pp.252-261
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• 2020

Objectives: Chronic diseases (CDs) continue to increase due to advances in medicine and increase in lifespan, affecting quality of life and resulting in economic loss through treatment costs. This study addressed the perception characteristics of the effectiveness of Korean medicine (KM) based on the existence of a CD to determine the national preference for KM treatment. Methods: Using data from the 2017 National Survey of KM Usage, we evaluated the perception on treatment effect of 16 diseases by dividing them into the CD group and the non-CD group. Response reliability was verified by applying the chi-square test (χ2-test) analysis method of the Statistical Package for the Social Sciences (SPSS) statistical program. Results: The analysis of the perception on effectiveness of KM for patients with CD (n = 1,050, 21.0%) and for patients with non-CD (n = 3,950, 79.0%) showed an overall similar trend for all the 16 diseases. The response rates of having some treatment effect were high for nine diseases namely, disc-related disease (CD: 70.7%, non-CD: 73.1%), osteoarthritis (CD: 72.3%, non-CD: 72.4%), frozen shoulder and shoulder pain (CD: 79.6%, non-CD: 81.4%), back pain (CD: 84.6%, non-CD: 85.0%), sprain (CD: 84.8%, non-CD: 84.1%), facial nerve paralysis (CD: 73.5%, non-CD: 71.7%), stroke (CD: 66.2%, non-CD: 62.8%), digestive disease (CD: 53.3%, non-CD: 50.0%), and common cold and rhinitis (CD: 44.7%, non-CD: 44.8%). Conclusion: The present results found that there was little difference in the perception on effectiveness of KM for each of the 16 diseases in patients with CD and non-CD, however, 70% or more of the respondents recognized some treatment effect on musculoskeletal disorders, regardless of the existence of CD. Preferential political support for KM treatment of chronic musculoskeletal disorders is recommended.

• 분석과학
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• 제15권2호
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• pp.163-171
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• 2002

수산가공 폐기물로 버려지는 게 껍질로부터 chitin을 추출하여 중금속 흡착제로 이용하였으며, 이 흡착제에 대해 Cd(II) 및 Pb(II) 이온의 흡착특성을 연구하였다. Chitin에 대한 Cd(II) 및 Pb(II) 이온의 흡착속도는 반응시간 2분경에 최대흡착량에 도달하였으며, 중금속 흡착에 미치는 pH의 영향은 두 이온 모두 pH 7.0>10.5>3.5순임을 알 수 있었다. Chitin에 대한 흡착률은 Cd(II)이온이 21${\sim}$99%이며, Pb(II)이온이 24${\sim}$95%이다. Cd(II)이온의 회수율은 22${\sim}$53%이고, Pb(II)이온의 회수율은 22${\sim}$73%로 나타났다. 이들 중금속 이온의 흡착양상은 Freundlich 흡착등온식에 비교적 잘 적용되었다.

• 대한전자공학회:학술대회논문집
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• 대한전자공학회 2000년도 하계종합학술대회 논문집(5)
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• pp.73-76
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• 2000

We study the Chua oscillator circuit by considering a negative resistor as a characteristic parameter which controlled by the CdS(Cadmium Sulphide) cell. This is a new way to observe several chaotic phenomena with the same initial condition continuously. Since we can control the internal resistance of the CdS cell by an additional circuit, our autonomous chaotic oscillator circuit makes it possible to measure the voltage value continuously and automatically.

• 분석과학
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• 제15권2호
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• pp.115-119
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• 2002

$\alpha$ 및 $\beta$-Cyclodextrin(CD)과 Congo red간의 복합체 형성에 관한 연구를 분광학적 방법으로 조사하였다. 이 결과 cavity 크기가 작은 $\alpha$-CD는 Congo red의 입체적 장애 때문에 복합체를 형성하지 못하였다. 한편 가시광선 영역의 S-형 흡광도 증가 현상으로 볼 때 두 분자의 $\beta$-CD가 한 분자의 Congo red와 equatorial 형태로 복합체를 형성하는 것 같다. 등 흡수점은 346 nm과 478 nm 두 곳에서 나타났다. 온도에 따른 형성상수의 값으로부터 온도가 올라가면 binding energy의 감소로 형성상수가 감소하였다. 열역학적 parameter의 계산 결과 $\Delta$H, $\Delta$S 및 $\Delta$G는 각각 -50.73 kJ/mol, $-108.96J/K{\cdot}mol$ 및 -18.24 kJ/mol 이었다. 따라서 큰 엔트로피 감소를 동반한 자발적 반응임을 알 수 있었다.

• 한국결정학회지
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• 제12권4호
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• pp.207-211
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• 2001

Cd(SCN)₂{CH/sub 6/H/sub 10/(NH₂)₂}₂(1) 착물을 합성하고 단결성 X-선 회절법으로 구조를 규명하였다. 이 착물은 단사정계, 공간국 P2₁/ space group with a=11.842(2), b=7.926(2), c=11.291(2) Å, β=106.73(3)°V=1014.8(4)Ų, Z=2 로 결정화되었으며, 1775 개의 독립적인 회절반점에 대한 최종 신뢰도 인자 R₁및 ωR₂값은 각각 0.0518 및 0.1315이었다. 착물의 결정 구조는 2개의1,2-Diaminocyclohexane리간드의 질소 원자 xy 평면에서 Cd(II) 금속원자에 chelate 되어 5-membered ring을 형성하며, 2개의 thiocyanate 리간드의 황 원자가 z 축 방향으로 Cd(II) 금속 원자에 트란스 형태로 결합함으로 써 z축 방향으로 약간 늘러진 팔면체 구조의 착물을 형성한다.