• Korean Journal of Food Science and Technology
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• v.47 no.2
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• pp.246-253
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• 2015

In this study, we evaluated the lipoprotein lipase (LPL) inhibitory activity of 11 water extracts derived from Cinnamomum cassia Blume, Sarcodon aspratus, Cordyceps militaris, Crataegus pinnatifida Bunge, Corni fructus, Allium cepa, Coix lacryma-jobi, Plantago asiatica L., Lentinus edodes, Rosa rugosa, and Foeniculum fructus. The results of the LPL secretion and activity assay showed Sarcodon aspratus (NE) extract have an LPL secretion inhibitory acitivity. The cause of reduction in LPL secretion after NE treatment was investigated using molecular biology methods. NE treatment affected the LPL content in cells, but did not affect LPL mRNA expression. It also increased the mRNA expression level of sortilin-related receptor LDLR class A (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL. Finally, cell fractionation revealed that NE treatment induced the expression of CCAAT-enhancer-binding protein beta ($C/EBP{\beta}$), a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. These results show that NE's anti-obesity effect involves inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.

• Journal of Life Science
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• v.15 no.1 s.68
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• pp.38-44
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• 2005

Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV, there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-1c1c7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-a, the inducibility of TNF-a expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-a expression. Infection of Hepa-1c1c7 cells with the serum of HBV carrier also increased TNF-a mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in $\beta-actin$ mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-lc1c7 in vitro and the viral infection induces TNF-a expression, which suggests that Hepa-lc1c7, a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.

• Proceedings of the Korean Information Science Society Conference
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• 2004.04b
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• pp.757-759
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• 2004

유전자 발현은 유전자가 mRNA와 생체의 기능을 일으키게 하는 단백질을 만들어내는 과정이다. 유전자 발현에 대한 정보는 유전자의 기능을 밝히고 유전자간의 상관 관계를 알아내는데 중요한 역할을 한다. 이러한 유전자 발현 연구를 위한 정보를 대량으로 신속하게 얻을 수 있는 도구가 DNA Chip이다. DNA Chip으로 얻은 수백-수천 개의 데이터는 그 데이터만으로는 의미를 갖지 못한다. 따라서 유전자 발현 정도에 따라 수치적으로 획득된 데이터에서 의미적인 특성을 찾아내기 위해서는 클러스터링 방법이 필요하다. 본 논문에서는 수많은 유전자 데이터 중에서 주요 정보를 포함한 것으로 판단되는 유전자 데이터를 선택하여 특징간을 계산하고 신경망 학습을 이용한 클러스터링하는 알고리즘에 대해서 기술한다.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.239-243
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• 2000

Helicobacter pylori, an etiologic agent of gastritis and peptic ulceration, produces urease which elicits a powerful immunoglobulin response in H. pylori-infected individuals. To establish a model plant vaccine agains H. pylori, 750 bp -ureA DNA amplified by polymerase chain reaction from pH 808 plasmid harboring urease gene cluster was cloned and manipulated to be expressed in tobacco plants. From the regenerated transgenic tobacco plants, ureA DNA integration,m its mRNA expression and protein synthesis were analyzed and confirmed by standard molecular techniques. The CaMV 35S promoter-driving ureA construct was expressed to produce a 30 kDa protein which was identical with bacterial UreA in size when detected on immunoblot of SDS polyacrylamide gel electrophoresis.

• Journal of Life Science
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• v.19 no.12
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• pp.1808-1814
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• 2009

In our previous study, a new herbal preparation (HemoHIM) was developed as a functional food for the radioprotection and immunomodulatory agents. In order elucidate the mechanism involved, we examined the effect of HemoHIM on the compound 48/80-induced histamine release, and on the phorbol 12-myristate 13-acetate (PMA)/calcium ionophore (A23187)-induced inflammatory cytokine secretion in HMC-1. The cell culture supernatants were harvested, and the cytokines (IL-4, IL-6, IL-8, TNF-$\alpha$, GM-CSF) in the supernatants were measured by enzyme-linked immunosorbent assay. The total RNA of the cells was extracted, and the cytokines or c-kit/tryptase/Fc$\varepsilon$RI's messenger RNA expressions were examined using reverse transcriptase polymerase chain reaction. Under low concentrations, HemoHIM inhibited histamine release in HMC-1 stimulated compound 48/80. Furthermore HemoHIM inhibited PMA/A23187-induced inflammatory cytokines' secreation or mRNA expression in a dose-dependent manner. But IL-8 secretion was not inhibited by low concentrayion of HemoHIM, respectively. The mRNA expression of c-kit and Fc$\varepsilon$RI were also inhibited in a dose-dependent manner. Tryptase mRNA expression was only inhibited by low concentration of HemoHIM. These results indicated that HemoHIM might be an useful agent for protection against allergy as well as immune modulation, especially since it is a relatively nontoxic natural product.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.25-25
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• 2003

어류의 번식활동은 뇌-뇌하수체-생식소로 이어지는 내분비기관이 주로 담당하고 있으며, 그 중에서 뇌하수체에서 만들어지는 생식선자극호르몬 (GTH)은 생식소 발달에 매우 중요한 역할을 한다. 한편, GTH는 생식소에서 생성되는 스테로이드호르몬에 의해 유전자의 발현이 조절된다. 본 연구에서는 산천어 (Oncorhynchus masou)의 GTH 유전자발현에 미치는 estradiol-17$\beta$ (E2)의 영향을 생체내에서 먼저 조사하고, 유전자발현조절의 메카니즘을 구명하기 위하여 에스트로젠 수용체 (ER)의 cDNA를 cloning하였다. 또한 내분비교란물질로 알려져 있으며 에스트로젠 수용체와의 유사결합작용이 알려진 노닐페놀 (NP)의 영향도 아울러 조사하였다. 미성숙 산천어를 E2 및 NP (각각 5 mg/kg 및 10 mg/kg 체중)로 처리하면 72 시간째에 뇌하수체내의 $\beta$-actin mRNA에 비해 GTH (FSH$\beta$, LH$\beta$) mRNA가 상대적으로 증가하는 경향을 나타내었다. 뇌하수체에서 분리한 ER의 기본적인 구조는 DNA결합도메인 (Zf-C4), 호르몬결합도메인 (hormone_rec), 기타 전사활성화도메인으로 구성되어 있었으며 (그림 1), 기존에 보고된 에스트로젠 수용체중에서 무지개송어 및 대서양연어의 ER과 각각 98%, 96%의 높은 상동성이 관찰되었다. 향후, 준비된 ER을 재료로 하여 GTH 유전자발현의 호르몬의존성을 조사할 예정이다.

• Development and Reproduction
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• v.4 no.1
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• pp.109-114
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• 2000

The present study was undertaken to examine the effects of ethanol on the ovarian steroidogenic acute regulatory protein(StAR) gene expression during prepubertal and onset of puberty. From day 25, each rat began receiving either a control saline or ethanol. Animals were sacrificed on day 27 and 32, and their ovaries and blood were collected. In the present results, ethanol treatment significantly decreased serum luteinizing hormone contents at both time points. Uterine weights of ethanol-treated group were significantly lighter than control group at early time point while there was no noticeable discrepancy at late time point. Vaginal openings, a marker of onset of puberty, also clearly delayed in ethanol-treated group. Using an in situ hybridization histochemistry, we determined the expression of mRNAs encoding StAR. Ovaries from ethanol-treated rats showed a suppresed expression of StAR mRNA. These results demonstrate that ethanol can disturb the prepubertal ovarian function and onset of puberty, at least in part, through the inhibition of ovarian StAR gene expression.

• Clinical and Experimental Pediatrics
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• v.52 no.5
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• pp.594-602
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• 2009

Purpose : Transforming growth factor (TGF)-${\beta}1$ reportedly increases neuronal survival by inhibiting the induction of inducible nitric oxide synthase (NOS) in astrocytes and protecting neurons after excitotoxic injury. However, the neuroprotective mechanism of $TGF-{\beta}1$ on hypoxic-ischemic (HI) brain injury in neonatal rats is not clear. The aim of this study was to determine whether $TGF-{\beta}1$ has neuroprotective effects via a NO-mediated mechanism and N-methyl-D-aspartate (NMDA) receptor modulation on perinatal HI brain injury. Methods : Cortical cells were cultured using 19-day-pregnant Sprague-Dawley (SD) rats treated with $TGF-{\beta}1$ (1, 5, or 10 ng/mL) and incubated in a 1% O2 incubator for hypoxia. Seven-day-old SD rat pups were subjected to left carotid occlusion followed by 2 h of hypoxic exposure (7.5% $O_2$). $TGF-{\beta}1$ (0.5 ng/kg) was administered intracerebrally to the rats 30 min before HI brain injury. The expressions of NOS and NMDA receptors were measured. Results : In the in vitro model, the expressions of endothelial NOS (eNOS) and neuronal NOS (nNOS) increased in the hypoxic group and decreased in the 1 ng/mL $TGF-{\beta}1-treated$ group. In the in vivo model, the expression of inducible NOS (iNOS) decreased in the hypoxia group and increased in the $TGF-{\beta}1$-treated group. The expressions of eNOS and nNOS were reversed compared with the expression of iNOS. The expressions of all NMDA receptor subunits decreased in hypoxia group and increased in the $TGF-{\beta}1$-treated group except NR2C. Conclusion : The administration of $TGF-{\beta}1$ could significantly protect against perinatal HI brain injury via some parts of the NO-mediated or excitotoxic mechanism.

• Journal of Veterinary Clinics
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• v.24 no.3
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• pp.295-299
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• 2007

The detrimental effects of gene transfection on embryo development and the molecular mechanism behind the differential expression of genes related to early embryo development were assessed in the production of transgenic cow embryos through somatic cell nuclear transfer (NT). Parthenogenetic, IVF, and transgenic NT embryos derived from ${\alpha}_1$-antitrypsin transfected ear fibroblast cells was produced. To investigate the molecular mechanism behind lower developmental competence of transgenic NT embryos, the differential mRNA expression of three genes ($IFN-{\tau}$, Oct4, Fgf4) in the 3 types of embryo (Parthenogenetic, IVF, transgenic NT) was examined. RNA was extracted from ten blastocysts derived from 3 types of embryos and reverse-transcripted for synthesis of the first cDNA. The quantification of 3 gene transcripts ($IFN-{\tau}$, Oct4, and Fgf4) was carried out in three replicate by quantitative real-time reverse transcriptase PCR. Expression level of $IFN-{\tau}$ mRNA was significantly higher in transgenic NT embryos than parthenogenetic and IVF embryos (P<0.05). However, expression level of Oct4 and Fgf4 of transgenic NT embryos was significantly lower than IVF embryos (P<0.05). Altered levels of these three mRNA transcripts may explain some of the embryonic/fetal/neonatal abnormalities observed in offspring from transgenic NT embryos.

• Journal of The Korean Society of Grassland and Forage Science
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• v.27 no.4
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• pp.241-248
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• 2007

Cytokinins are essential plant hormones that play crucial roles in various aspects of plant growth and development. By using mRNA differential display, we isolated a cytokinine-inducible cDNA encoding pathogenesis-related (PR) 4 from Arabidopsis amp1 mutant. The full-length PR4 cDNA, designated AtPR4, contains an open reading frame of 212 amino acids with calculated molecular mass of 22,900 Da and isoelectric point (pI) of 7.89. Genomic DNA blotting showed that the Arabidopsis genome has one copy of AtPR4. AtPR4 mRNA was induced by cytokinin and NaCl, but decreased by SA or JA treatment. PR proteins are induced in response to pathogen attack. Thus the AtPR4 gene isolated in this study may be a useful candidate for genetic engineering of forage crops for increased tolerance against pathogen.