• Journal of Life Science
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• v.29 no.6
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• pp.724-734
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• 2019

The present study investigated the cytotoxicity of particulate matter (PM) derived from car air filter (outdoor PM) and home cleaner filter (indoor PM) in the various human cell lines. Each outdoor and indoor PM were harvested by ethanol extraction method, subsequently sieved with 10 um filter paper, sterilized with autoclave and added to culture media. The half maximal inhibitory concentration ($IC_{50}$) values was significantly (p<0.05) lower in the outdoor PM, compared with indoor PM, and the significantly (p<0.05) higher $IC_{50}$ values were observed in the cancer cell lines (A-549 lung adenocarcinoma and AGS stomach adenocarcinoma), than those of normal MRC-5 fibroblasts and dental papilla tissue derived-mesenchymal stem cells (DSC). After being exposed to $100{\mu}g/ml$ outdoor PM for 7 days, the population doubling time (PDT) was significantly (p<0.05) increased in especially MRC-5 and DSC cell lines, compared with untreated cell lines. Further, the expression of senescence-associated ${\beta}$-galactosidase activity was up-regulated in all the cells exposed to outdoor PM than those of untreated control. Besides, the expression level of inflammation-associated genes, such as cyclooxygenase-2 (COX-2) and interleukin-6 (IL-6) was found to be significantly (p<0.05) increased in the outdoor PM-treated cell lines than those of untreated cell lines. Our results showed that PM induces the cytotoxicity via arrest of cell growth, cell damage and inflammation response.

• Journal of Nutrition and Health
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• v.55 no.2
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• pp.227-239
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• 2022

Purpose: This study investigated the effects of water-soluble mulberry leaf extract (ME) on hepatic lipid accumulation in high-fat diet-fed rats via the regulation of hepatic microRNA (miR)-221/222 and inflammation. Methods: Male Sprague-Dawley rats (4 weeks old) were randomly divided into 3 groups (n = 7 each) and fed with 10 kcal% low-fat diet (LF), 45 kcal% high-fat diet (HF), or HF + 0.8% ME for 14 weeks. Lipid profiles and cytokine levels of the liver and serum were measured using commercial enzymatic colorimetric and enzyme-linked immunosorbent assay, respectively. The messenger RNA (mRNA) and miR levels in liver tissue were assayed by real-time quantitative reverse-transcription polymerase chain reaction. Results: Supplementation of ME reduces body weight and improves the liver and serum lipid profiles as compared to the HF group. The mRNA levels of hepatic peroxisome proliferator-activated receptor-gamma, sterol regulatory element binding protein-1c, fatty acid synthase, and fatty acid translocase, which are genes involved in lipid metabolism, were significantly downregulated in the ME group compared to the HF group. In contrast, the mRNA level of hepatic carnitine palmitoyl transferase-1 (involved in fatty acid oxidation) was upregulated by ME supplementation. Furthermore, administration of ME significantly downregulated the mRNA levels of inflammatory mediators such as hepatic tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), monocyte chemoattractant protein-1, and inducible nitric oxide synthase. The serum levels of TNF-α, IL-6, and nitric oxide were also significantly reduced in ME group compared to the HF group. Expression of hepatic miR-221 and miR-222, which increase in the inflammatory state of the liver, were also significantly inhibited in the ME group compared to the HF group. Conclusion: These results indicate that ME has the potential to improve hepatic lipid accumulation in high-fat diet-fed rats via modulation of inflammatory mediators and hepatic miR-221/222 expressions.

• Journal of Dental Rehabilitation and Applied Science
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• v.40 no.1
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• pp.13-23
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• 2024

Purpose: This aim of this study was to demonstrate growth factors that differentiate human mesenchymal stem cells into chondrocytes and to evaluate cell proliferation enhancement by needle size differences. Materials and Methods: Human mesenchymal stem cells were cultured in chondrogenic medium supplemented with BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1, TGF-β2, TGF-β3 and without growth factors for 14, 21, and 28 days. Then, the expression levels of SOX-5, SOX-6, SOX-9 and FOXO1A were comparatively analyzed. Human mesenchymal stem cells were inoculated into culture dishes using 18, 21, and 26 gauge (G) needles, and cell proliferation was measured after 24, 48, and 72 hours, respectively. Results: In addition to the previously known FGF, IGF-1, and TGFβ1,the BMP family growth factors such as BMP-2, BMP-4, BMP-6, and BMP-7 increased the expression of chondrocyte differentiation genes SOX-5, SOX-6, SOX-9, and FOXO1A. At 48 hours, the 26G group, the smallest needle, showed significant cell proliferation improvement compared to the control group and the 18G group. At 72 hours, the 26G group, the smallest needle, showed significant increase in cell proliferation compared to the control group. Conclusion: Through this study, growth factors with the ability to induce chondrocyte differentiation of human mesenchymal stem cells were investigated, and cell proliferation changes by needle size differences were determined.

• Clinical and Experimental Pediatrics
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• v.48 no.4
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• pp.380-386
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• 2005

Purpose : The incidence of nonphysiologic neonatal hyperbilirubinemia is twice as high in East Asians as in whites. Recently, UGT1A1 mutation was found to be a risk factor for neonatal hyperbilirubinemia. In congenitally-jaundiced Gunn rats, which lack expression of UDP-glucuronosyltransferase, alternative pathways can be stimulated by inducers of CYP1A1 and CYP1A2 enzymes. CYP1A2 plays a major role in bilirubin degradation of the alternate pathway. We studied the relationship between UGT1A1 and CYP1A2 gene polymorphism of neonatal hyperbilirubinemia in Koreans. Methods : Seventy-nine Korean full term neonates who had hyperbilirubinemia(serum bilirubin >12 mg/dL) without obvious causes of jaundice, were analyzed for UGT1A1 and CYP1A2 gene polymorphism; the control group was sixty-eight. We detected the polymorphism of Gly71Arg of UGT1A1 gene by direct sequencing and T2698G of CYP1A2 by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) using MboII and direct sequencing. Results : Allele frequency of Gly71Arg mutation in the hyperbilirubinemia group was 32 percent, which was significantly higher than 11 percent in the control group(P<0.0001). Mutant gene frequency of T2698G was 41.8 percent in patients and 32.3 percent in the control group(P=0.015), but allele frequency was 21 percent in patients and 19 percent in the control group, which was not significantly higher(P=0.706). There was no relationship between mutations of two genes(P=0.635). Conclusion : The polymorphism of UGT1A1 gene(Gly71Arg) and CYP1A2 gene(T2698G) was detected in Korean neonatal hyperbilirubinemia. Only polymorphisms of Gly71Arg in UGT1A1 were significantly higher than control group.

• Tuberculosis and Respiratory Diseases
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• v.62 no.5
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• pp.406-416
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• 2007

Background: Single nucleotide polymorphisms (SNPs), which consist of a substitution of a single nucleotide pair, are the most abundant form of genetic variations occurring with a frequency of approximately 1 per 1000 base pairs. SNPs by themselves do not cause disease but can predispose humans to disease, modify the extent or severity of the disease or influence the drug response and treatment efficacy. Single nucleotide polymorphisms (SNPs), particularly those within the regulatory regions of the genes often influence the expression levels and can modify the disease. Studies examining the associations between SNP and the disease outcome have provided valuable insight into the disease etiology and potential therapeutic intervention. Traditionally, the genotyping of SNPs has been carried out using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP), which is a low throughput technique not amenable for use in large-scale SNP studies. Recently, TaqMan real-time PCR chemistry was adapted for use in allelic discrimination assays. This study validated the accuracy and utility of real-time PCR technology for SNPs genotyping Methods: The SNPs in promoter sequence (-37 and -524) of lung cancer suppressor gene, RRM1 (ribonucleotide reductase M1 subunit) with the genomic DNA samples of 89 subjects were genotyped using both real-time PCR and PCR-RFLP. Results: The discordance rates were 2.2% (2 mismatches) in -37 and 16.3% (15 mismatches) in -524. Auto-direct sequencing of all the mismatched samples(17 cases) were in accord with the genotypes read by real-time PCR. In addition, 138 genomic DNAs were genotyped using real-time PCR in a duplicate manner (two separated assays). Ninety-eight percent of the samples showed concordance between the two assays. Conclusion: Real-time PCR allelic discrimination assays are amenable to high-throughput genotyping and overcome many of the problematic features associated with PCR-RFLP.

• Radiation Oncology Journal
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• v.21 no.3
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• pp.222-226
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• 2003

Purpose: The interaction of the Fas: Fas ligand has been recognized to play an important role in radiation induced apoptosis. The purpose of this study was to investigate the role of Fas and Fas ligand mutations, in radiation-induced apoptosis in vivo. Materials and Methods: Mice with a mutation in the Fas ($C57BL/6J-Fas^{lpr}$) and its normal control (C57BL/6J) and the Fas ligand ($C3H/HeJ-Fas^{gld}$) and its normal control (C3H/HeJ), were used in this study. Eight-week old male mice were given whole body radiation. After irradiation, the mice were killed at various time intervals, and their spleens collected. Tissue sample was stained with hematoxylin-eosin, and the numbers of apoptotic cells scored. The regulating molecules of apoptosis including the p53, Bcl-2, Bax, $Bcl-X_L\;and\;Bcl-X_s$ genes were also analyzed by Western blotting. Results: With 2.5 Gy and 10 Gy of irradiation, the levels of apoptosis were lower in the $C57BL/6J-Fas^{lpr}\;and\;C3H/HeJ-Fas^{gld}$ mice than in the control mice (p<0.05). With the expression of apoptosis regulating molecules, the Bax was increased in both the C57BL/6J and C3H/HeJ mice in response to radiation; the peak levels of Bax in the C57BL6J and C3H/HeJ were 3 and 3.3-fold higher after 8hr, respectively. However the Bax was not increased in either the $C57BL/6J-Fas^{lpr}\;or\;C3H/HeJ-Fas^{gld}$mice. The p53, Bcl-X_L,\;Bcl-X_S$and Bcl-2 showed no significant changes in the $C57BL/6J-Fas^{lpr},\;C3H/HeJ-Fas^{gld}$, C57BL/6J and C3H/HeJ mice. Conclusion: The levels of radiation-induced apoptosis were lower in the lpr and gld, than the control mice, which seemed to be related to the level of Bax activation due to the radiation in the lpr and gld mice. This result suggests that Fas/Fas L plays an important role in radiation-induced apoptosis in vivo.

• Development and Reproduction
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• v.6 no.2
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• pp.131-139
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• 2002

Since GnRH and its receptor genes are expressed in the ovary, it has been suggested that ovarian GnRH might be involved in the regulation of ovarian function and the apoptosis of ovarian cells. However, it was not known well on the expression and function of GnRH and its receptor in the corpus luteum. The present study was undertaken to investigate whether GnRH and its receptor are expressed in luteal cells and GnRH has any effect on the apoptosis of luteal cells. Luteal cells obtained from the pregnant rats were cultured and stained for GnRH and its receptor proteins. Cultured luteal cells showed distinct immunoreactivity against both anti-GnRH and anti-GnRH receptor antibodies. In addition, the presence of GnRH receptor protein in cultured cells was confirmed by Western blot analysis. To investigate the effect of GnRH on the apoptosis of luteal cells, luteal cells were cultured in the presence of 10$^{-6}$ M GnRH-agonist(GnRH-Ag) for 3, 8, and 12h. TUNEL assay showed that the number of cells undergoing apoptosis increased 12h after culture(P<0.05). DNA fragmentation analysis confirmed the results such that the cells treated for 12h showed the greatest increase of fragmentation(p<0.05). Further, Western blot analysis of cytochrome c in the mitochondrial and cytoplasmic fractions of the luteal cells showed that GnRH-Ag treatment increased the content of cytochrome c in cytoplasm. These results demonstrate that the luteal cells express GnRH and its receptor and GnRH-Ag treatment induces apoptosis of the luteal cells via mitochondrial release of cytochrome c. The present study suggest that the releasing of cytochrome c from mitochondria might be involved in the luteal cell apoptosis induced by GnRH-Ag.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.65 no.4
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• pp.406-415
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• 2020

Maize (Zea mays L.) is one of the most valuable agricultural crops and is grown under a wide spectrum of environmental conditions. However, maize is moderately sensitive to salt stress, and soil salinity is a serious threat to its production worldwide. In this study, we used ethyl methane sulfonate (EMS) to generate salt-tolerant silage maize mutants. We screened salt-tolerant lines from 203 M₃ mutant populations by evaluating the morphological phenotype after salt stress treatment and selected the 140ES91 line. The 140ES91 mutant showed improved plant growth as well as higher proline content and leaf photosynthetic capacity compared with those of wild-type plants under salt stress conditions. Using whole-genome re-sequencing analysis, 1,103 single nucleotide polymorphisms and 71 insertions or deletions were identified as common variants between KS140 and 140ES91 in comparison with the reference genome B73. Furthermore, the expression patterns of three genes, which are involved in salt stress responses, were increased in the 140ES91 mutant under salt stress. Taken together, the mutant line identified in our study could be used as an improved breeding material for transferring salt tolerance traits in maize varieties.

• Journal of Life Science
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• v.21 no.5
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• pp.753-760
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• 2011

In the present study, the effects of paprika extract and its components including vitamin C, lycopene and beta-carotene on cell death in ultraviolet B (UVB)-exposed HaCaT cells were investigated. The cell viability upon treatment for 24 hr with either paprika extract or vitamin C alone was similar to or greater than that of the untreated control. However, the viability of the cells treated with lycopene or beta-carotene decreased to about 20% of that in the untreated control. When UVB-exposed cells were post-incubated for 24 hr in medium containing paprika extract or vitamin C, cell viability increased in a concentration dependent manner as compared to those post-incubated in a normal growth medium. In contrast, post-incubation of UVB-exposed cells with lycopene or beta-carotene decreased cell viability in a concentration dependent manner as compared to those post-incubated in a normal growth medium. The nuclear fragmentation analysis showed that paprika extract or vitamin C decreases UVB-induced apoptosis. The apoptotic nuclear fragmentation resulting from UVB exposure was also protected by the paprika extract or vitamin C post-incubation. However, the UVB-induced apoptotic nuclear fragmentation of the cells treated with lycopene or beta-carotene increased in a concentration dependent manner. Western blot analysis showed that either paprika extract or vitamin C treatment alone did not significantly change the level of p53 and GADD45 protein. Interestingly, post-incubation of UVB-exposed cells with paprika extract or vitamin C decreased the p53 and GADD45 protein level as compared to those post-incubated in a normal growth medium. In contrast, incubation of UVB-exposed or non-irradiated cells with lycopene or beta-carotene increased the p53 and GADD45 protein levels in a concentration dependent manner as compared to those incubated in a normal growth medium. All these results suggest that paprika extract and vitamin C help the survival of the UVB-exposed cells, while lycopene and beta-carotene potentiate the apoptotic death of UVB-exposed cells, in accordance with the respective changes in p53 and GADD45 protein levels.

• Development and Reproduction
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• v.15 no.2
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• pp.179-185
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• 2011

Some phytoestrogens in soy and red wine, for example, might have beneficiary rather than adverse effects. In particular, dietary soy intake seems to be highly correlated with protection of breast cancer, osteoporosis and cardiovascular disorders. However, questions persist on the potential adverse effects of the main soy constituent genistein (GS) on female reproductive physiology. Previously we found that prepubertal exposure to GS could activate the reproductive system of immature female rats leading to precocious puberty onset, and intracerebroventricularly (ICV) injected GS could directly activate hypothalamic kisspeptin-GnRH neuronal circuits in adult female rats. The present study was performed to examine the hypothalamus-specific GS effects in prepubertal female rats and which subtype of estrogen receptor is mediated in this GS effect. Prepubertal female rats (PND 30) were anaesthetized, treated with single dose of GS (3.4 ${\mu}g$/animal), and sacrificed at 2 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly lowered the transcriptional activities of mTOR (1:$0.361{\pm}0.058$ AU, p<0.001) but increased that of GAD67 (1:$1.285{\pm}0.099$ AU, p<0.05), which are known to act as an upstream modulator of kisspeptin and GnRH neuronal activities in the hypothalamus, respectively. GS administration enhanced significantly the mRNA levels of KiSS-1(1:$1.458{\pm}0.078$ AU, p<0.001), and exerted no effect on the mRNA level of kisspeptin receptor GPR-54 (1:$1.29{\pm}0.08$ AU). GnRH gene expression was significantly decreased in GS-treated group compared to control group (1:$0.379{\pm}0.196$ AU, p<0.05). There was no difference in the mRNA level of $ER{\alpha}$ in the GS-treated group compare to control group (1:$1.180{\pm}0.390$ AU, Fig. 3A). However, icv infusion of GS significantly increased the transcriptional activities of $ER{\beta}$ (1:$4.209{\pm}0.796$ AU, p<0.01, Fig. 3B). Taken together, the present study indicated that the acute exposure to GS could directly alter the hypothalamic GnRH modulating system in prepubertal female rats. Our study strongly suggested the involvement of $ER{\beta}$ pathway in GS's hypothalamus-specific action, and this idea is consistent with the GS's well-known $ER{\beta}$-mediated protective action in breast cancer.