• Journal of Embryo Transfer
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• v.31 no.3
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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.

• Journal of Embryo Transfer
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• v.33 no.3
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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.

• Development and Reproduction
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• v.21 no.2
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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.

• Journal of the korean academy of Pediatric Dentistry
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• v.46 no.3
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• pp.337-342
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• 2019

The aim of this study was to analyze cells from human dental pulp tissue of impacted supernumerary teeth as stem cells with flow cytometry. Human dental pulp cells from 15 supernumerary teeth were identified their characteristics as stem cells by expression of mesenchymal stem cell markers through flow cytometry analysis at passage 3 and passage 10. Cluster of differentiation (CD) 73, CD 90, CD 34, CD 45 and STRO-1 cell surface markers were used to figure out characteristics of dental pulp stem cells from supernumerary teeth. At passage 3, the cell population showed positive expression of CD 73, CD90 and STRO-1, lacked expression of CD 34 and CD 45. At passage 10, CD 73, CD 90 and STRO-1 showed positive expression while CD 34 and CD 45 showed negative expression. This study indicated that dental pulp stem cells of supernumerary teeth had the properties of mesenchymal stem cells at both early and late passage. Impacted supernumerary teeth could be considered as a noble source of stem cells because of rapid growth and maintaining characteristics of stem cells until late passage.

• Proceedings of the Materials Research Society of Korea Conference
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• 2001.11a
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• pp.73-73
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• 2001

• Journal of Pharmacopuncture
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• v.23 no.4
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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.

• Analytical Science and Technology
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• v.15 no.2
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• pp.163-171
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• 2002

The adsorption characteristics of Cd(II) and Pb(II) ions has been studied by using chitin as an adsorbent. The pure chitin was obtained from the extraction of red-crab shell dumped by fish factory. Adsorption kinetics of Cd(II) and Pb(II) ions on the chitin reached at the maximum adsorption within two minutes. Adsorbed amounts of heavy metals were pH 7.0>10.5>3.5 in the following order. Adsorption ratio by chitin was 21${\sim}$99% for Cd(II) ion and 24${\sim}$95% for Pb(II) ion. Recovery ratio of Cd(II) ion on the chitin was 22${\sim}$53%, and that of Pb(II) ion was 22${\sim}$73%. The adsorption behavior of these heavy metals was explained well by Freundlich adsorption isotherm.

• Proceedings of the IEEK Conference
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• 2000.06e
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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.

• Analytical Science and Technology
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• v.15 no.2
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• pp.115-119
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• 2002

The formation of inclusion complexes between cyclodextrin(CD) and Congo red was studied by spectrophotometric methods at various temperatures. The cavity sizes are 0.49 nm, 0.62 nm for $\alpha$-and $\beta$-CD, respectively. Therefore, $\alpha$-CD was not found to form an inclusion complex with Congo red due to steric hinderance. In the $\beta$-CD use two $\beta$-CD molecules formed an inclusion complex with one molecule of Congo red, from the slope of the S-shaped curve increased. Two prominent isosbestic points appear at 346 nm and 478 nm. The formation constants were decreased with the increasing temperatures, due to low binding energy between $\beta$-CD and Congo red. The thermodynamic parameters were calculated from the plot of $lnK_f$ vs 1/T. The $\Delta$H, $\Delta$S and $\Delta$G were -50.73 kJ/mol, $-108.96J/K{\cdot}mol$ and -18.26 kJ/mol, respectively.

• Korean Journal of Crystallography
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• v.12 no.4
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• pp.207-211
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• 2001

The crystal structure of the title complex, Cd(SCN)₂{CH/sub 6/H/sub 10/(NH₂)₂}₂(1) has been analyzed by X-ray single crystallography. The complex 1 crystallizes in the monoclinic system 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, R₁=0.0518 and ωR₂=0.1315 for 1775 independent reflections. The central Cd(II) atom of this com-plex has a slightly distorted octahedral coordination geometry, with the 1,2-Diaminocyclohexane ligands functioning as an N,N'-bidentate and the thiocyanate ligands bonding through the sulfur atom in a trans arrangement.