• Molecules and Cells
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• v.25 no.2
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• pp.265-271
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• 2008

Single nucleotide polymorphisms (SNPs) are the most common form of human genetic variation. Non-synonymous SNPs (nsSNPs) change an amino acid. Organic anion transporters (OATs) play an important role in eliminating or reabsorbing endogenous and exogenous organic anionic compounds. Among OATs, hOAT4 mediates high affinity transport of estrone sulfate and dehydroepiandrosterone sulfate. The rapid bone loss that occurs in post-menopausal women is mainly due to a net decrease of estrogen. In the present study we searched for SNPs within the exon regions of hOAT4 in Korean women osteoporosis patients. Fifty healthy subjects and 50 subjects with osteoporosis were screened for genetic polymorphism in the coding region of SLC22A11 (hOAT4) using GC-clamp PCR and denaturing gradient gel electrophoresis (DGGE). We found three SNPs in the hOAT4 gene. Two were in the osteoporosis group (C483A and G832A) and one in the normal group (C847T). One of the SNPs, G832A, is an nsSNP that changes the $278^{th}$ amino acid from glutamic acid to lysine (E278K). Uptake of [$3^H$] estrone sulfate by oocytes injected with the hOAT4 E278K mutant was reduced compared with wild-type hOAT4. Km values for wild type and E278K were $0.7{\mu}M$ and $1.2{\mu}M$, and Vmax values were 1.8 and 0.47 pmol/oocyte/h, respectively. The present study demonstrates that hOAT4 variants can causing inter-individual variation in anionic drug uptake and, therefore, could be used as markers for certain diseases including osteoporosis.

• Journal of Animal Science and Technology
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• v.61 no.2
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• pp.61-68
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• 2019

The hG-CSF (human Granulocyte Colony-Stimulating Factor) is a growth and stimulation factor capable of inducing the proliferation of bone marrow cells, several types of leukocytes, among other hematopoietic tissue cells. hG-CSF is used in used to treat anomalies that reder a small number of circulating white blood cells, which may compromise the immune defenses of the affected person. For these reasons, the production of hG-CSF in a bioreactor system using the mammary gland of genetic modified animals is a possibility of adding value to the bovine genetic material and reducing the costs of hG-CSF production in pharmaceutical industry. In this study, we aimed the production of transgenic hG-CSF bovine through the lipofection of bovine primary fibroblasts with an hG-CSF expression cassette and cloning these fibroblasts by the somatic cell nuclear transfer (SCNT) technique. The bovine fibroblasts transfected with the hG-CSF cassette presented a stable insertion of this construct into their genome and were efficiently synchronized to G0/G1 cell cycle stage. The transgenic fibroblasts were cloned by SCNT and produced 103 transferred embryos and 2 pregnancies, one of which reached 7 months of gestation.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.65-69
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• 1992

The sucess of intrauterine inseminations with washed human spermatozoa was evaluated in 92 patients. In according to indication of insemination, intrauterine inseminations of women with hostile cervical mucus yield a 35.3% and 12.8% with male factors and 27.3% with unexplained infertility. In ovulation induction group with variable agents including clomiphen, human menopausal gonadotropin (HMG) and GnRH analog, the pregnancy rate was 27.8% (22/79) and in natural cycle group, 15.4% (2/13). The fetal loss rate in insemination group was 12.5% (3/24). Multiple pregnancies were 7 cases and ovulation induction were performed in 6 cases among them. Intrauterine insemination with washed human spermatozoa therefore represents an effective and safe procedure selected infertile couples.

• Journal of the Korean Magnetic Resonance Society
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• v.7 no.1
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• pp.37-45
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• 2003

Human CD99 is a ubiquitous 32-kDa transmembrane protein encoded by mic2 gene. Recently it has been reported that expression of a splice variant of CD99 transmembrane protein (Type I and Type II) increases invasive ability of human breast cancer cells. To understand structural basis for cellular functions of CD99 Type II, we have initiated studies on hCD99$\^$TMcytoI/ using circular dichroism (CD) and multi-dimensional NMR spectroscopy. CD spectrum of hCD99$\^$TMytoI/ in the presence of 200mM DPC and CHAPS displayed an existence ${\alpha}$-helical conformation, showing that it could form an ${\alpha}$-helix under membrane environments. In addition, we have found that the cytoplasmic domain of CD99 would form symmetric dimmer in the presence of transmembrane domain. Although it has been rarely figured out the correlation between structure and functional mechanism of hCD99$\^$TMcytoI/, the dimerization or oligomerization would play an important role in its biological function.

• Journal of the East Asian Society of Dietary Life
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• v.12 no.4
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• pp.289-298
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• 2002

This study was designed to investigate the anti-tumor effects of fresh carrot juice, methanol-extracts, and $\beta$-carotene on the human lung cancer cell line NCI-H1299. The anti-tumor effect was evaluated by the MTT assay in vitro. The anti-tumor effect of fresh carrot juice against NCI-H1299 lasted up to 96 hours after exposure; the viability rate of lung cancer cells decreased below 50% after 48 hours, and further after 72 hours. The strongest propagation inhibition effect of fresh carrot juice was shown at the concentration of 2000 $\mu\textrm{g}$/$m\ell$ after 72 hours and the viability rates was 45.98% even at the concentration of 25 $\mu\textrm{g}$/$m\ell$. The value of $IC_{50}$/ was 23.1$\mu\textrm{g}$/$m\ell$ when the elapsed time was 72 hours. The viability rate of methanol-extract was 52.4% under the concentration of 2000 $\mu\textrm{g}$/$m\ell$ and the elapsed time of 72 hours. Under the concentration of 1000 $\mu\textrm{g}$/$m\ell$ and the elapsed time of 48 hours, $\beta$ -carotene decreased the viability rate to 29.99%. The $IC_{50}$/ value of $\beta$-carotene was 691.2$\mu\textrm{g}$/$m\ell$ after 72 hours. According to the above results, the anti-tumor effect arose in NCI-H1299 when the concentration of the fresh carrot juice or the $\beta$-carotene was more than 25 $\mu\textrm{g}$/$m\ell$ or 1000 $\mu\textrm{g}$/$m\ell$, respectively. On the other hand, the methanol-extracts showed a weak anti-tumor effect even at a concentration as high as 2000 $\mu\textrm{g}$/$m\ell$.

• BMB Reports
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• v.35 no.4
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• pp.395-402
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• 2002

Caveolae are small, flask-shaped, non-clathrin coated invaginations of the plasma membrane of many mammalian cells. Caveolae have a coat that includes caveolin. They have been implicated in numerous cellular processes, including potocytosis. Since the human folate receptor (hFR) and other glycosyl-phosphatidylinositol (GPI)-tailed proteins have been co-localized to caveolae, we studied the caveolin role in the hFR function by transfecting hFR and/or caveolin cDNA into Fischer rat thyroid epithelial (FRT) cells that normally do not express detectable levels of either protein. We isolated and characterized stable clones as follows: they express (1) high levels of caveolin alone, (2) hFR and caveolin, or (3) hFR alone. We discovered that hFR is correctly processed, sorted, and anchored by a GPI tail to the plasma membrane in FRT cells. No difference in the total folic acid binding or cell surface folic acid binding activity were found between the FRT cells that were transfected with hFR, or cells that were transfected with hFR and caveolin. The hFR that was expressed on the cell surface of clones that were transfected with hFR was also sensitive to phosphatidylinositol-specific phospholipase C (PI-PLC) release, and incorporated radiolabeled ethanolamine that supports the attachment of a GPI-tail on hFR. We conclude that the processing, sorting, and function of hFR is independent on the caveolin expression in FRT cells.

• Journal of Ginseng Research
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• v.22 no.2
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• pp.140-146
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• 1998

It was founded that an X-compound is contained in extracts from the root of old red ginseng (6RG) compared with that from the root of 4-year old red ginseng(4RG). Both water extract and petroleum ether extract (PEII) from 6RG or 4RG inhibited the release of [$^{3}H$]-serotonin induced by platelet activating factor (PAF; 40 ng/ml). Water extract and PEll from 6RG Inhibited potently PAF-induced [$^{3}H$]-serotonin release compared with those from 4RG. X-compound out of both water extract and PEll from 6RG inhibited the release of [$^{3}H$]-serotonin inducted by collagen (100 $\mu\textrm{g}$/ml) or thrombin(20 U/ml). X-compound had a synergistic effect with water extract from 4RG on collagen-and thrombin-induced [3H] -serotonin release out of human platelets. The concentration(IC50) of X-compound that require to inhibit 50% of [$^{3}H$]-serotonin-release was 3.25 $\mu\textrm{g}$/ml, and it is inferred that maximum concentration of X-compound that inhibits the release of [$^{3}H$]-serotonin is 10 $\mu\textrm{g}$/ml. Because thrombosis is resulted mainly from the irreversible aggregations which are intimately related with the serotonin release and migraine is also caused when serotonin is released, it is inferred that water extract, PEII and X-compound from 6RG have antithrombosis and antimigrainous functions by inhibiting the release of serotonin from human platelets.

• The Korean Journal of Nuclear Medicine
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• v.38 no.1
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• pp.99-108
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• 2004

Purpose: The ability to noninvasively track the migration of neural progenitor cells would have significant clinical and research implications. We generated stably transfected F3 human neural progenitor cells with human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation. Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent (5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to $20{\mu}M$, up to 62.5nM, respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell passage. Therefore, the gene ex[ressopm of exogenous gene transferred human stem cell might be affected to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture conditions.

• Bulletin of the Korean Chemical Society
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• v.30 no.9
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• pp.2083-2086
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• 2009

Human vascular endothelial growth factor receptor 2 (hVEGFR2) is an important signaling protein involved in angiogenesis and attractive drug target in cancer therapy. It has been reported that flavonols, a class of flavonoids, have anti-angiogenic activity in various cancer cell lines. We performed receptor-oriented pharmacophore based in silico screening for identification of hVEGFR2 inhibitors from flavonol database. By comparing with three X-ray complex structures of hVEGFR2 and its inhibitors, we evaluated the specific interactions between inhibitors and receptors and determined a single pharmacophore map. This map consisted of four features, a hydrogen bonding acceptor (HBA) on Cys917, two hydrogen bonding donors on Glu917 (HBD1) and Glu883 (HBD2), and one hydrophobic interaction (Lipo) with Val846, Ala864, Val897, Val914 and Phe1045 of hVEGFR2. Using this map, we searched a flavonol database including 9 typical flavonols and proposed that five flavonols, kaempferol, quercetin, fisetin, morin, and rhamnetin can be potent inhibitors of hVEGFR2. 3-OH of C-ring and 4’-OH of B-ring of flavonols are the essential features for hVEGFR2 inhibition. This study will be helpful for understanding the mechanism of inhibition of hVEGFR2 by natural products.

• BMB Reports
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• v.47 no.10
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• pp.563-568
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• 2014

Human genome projects have enabled whole genome mapping and improved our understanding of the genes in humans. However, many unknown genes remain to be functionally characterized. In this study, we characterized human chromosome 4 open reading frame 34 gene (hC4orf34). hC4orf34 was highly conserved from invertebrate to mammalian cells and ubiquitously expressed in the organs of mice, including the heart and brain. Interestingly, hC4orf34 is a novel ER-resident, type I transmembrane protein. Mutant analysis showed that the transmembrane domain (TMD) of hC4orf34 was involved in ER retention. Overall, our results indicate that hC4orf34 is an ER-resident type I transmembrane protein, and might play a role in ER functions including $Ca^{2+}$ homeostasis and ER stress.