• Journal of the East Asian Society of Dietary Life
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• v.7 no.4
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• pp.484-490
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• 1997

In an attempt to investigate the effects of white ginseng on the proliferation and the nitric oxide(NO) secretion of mouse peritoneal macrophages, which are the first mai or defense phagocytes in the immune system, the studies have been carried out. In the macrophage proliferation assay using the $^3$H-thymidine incorporation, the total saponin or Ginsenoside Rb$_2$ were added to the medium at the concentration of 0 to 256$\mu\textrm{g}$/$m\ell$. DNA synthesis of the macrophage was increased at 64$\mu\textrm{g}$/$m\ell$ of total saponin and either 16$\mu\textrm{g}$/$m\ell$ or 64$\mu\textrm{g}$/$m\ell$ of Ginsenoside Rb$_2$, respectively. Also, the effect o(white ginseng on the nitric oxide secretion of the macrophages was investigated. The addition of either total saponin or Ginsenoside Rb$_2$ at the concentration of 20$\mu\textrm{g}$/$m\ell$ significantly increased the secretion of NO from the macrophages. The nitric oxide synthase (NOS) gene expression which is responsible for the synthesis of the nitric oxide has been studied using reverse transcription polymerase chain reaction. In RT-PCR, the $\beta$-actin and nos gene expression have been analyzed. 20$\mu\textrm{g}$/$m\ell$ of either total saponin or Ginsenoside Rb$_2$ increased nos gene expression of the macrophages.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.80-80
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• 2003

Insulin-dependent or Type 1 diabetes mellitus (IDDM) has been associated with an increased severity of periodontal disease. Since periodontal ligament (PDL) cells play a significant role in maintenance and regeneration of mineralized tissue, the success of procedures, such as guided tissue regeneration, is directly related to the ability of these cells to augment mineralized tissue. In this study, we investigated the time- and dose-dependent effect of high glucose on the proliferation and collagen synthesis of human periodontal ligament (PDL) cells. PDL cells were treated with high glucose (22mM, 33mM, 44mM) for 1 or 2 days. High glucose significantly inhibited proliferation of PDL cells as a time- and dose-dependent manner as evidenced by MTT assay. PDL cells were cultured in high glucose media (22mM, 33mM, 44mM) for 24 h. The ratio of collagen content to total protein was evaluated, and the gene expression of type I collagen was assessed by RT - PCR. The high concentration of glucose inhibited collagen synthesis, a marker of bone formation activity. This study indicated high glucose concentration could alter the metabolism of periodontal ligament cell, leading to alveolar bone destruction.

• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.60-73
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• 2005

Objectives : The aim of this study was to evaluate the effects of Loranthus parasiticus Merr. on cell cycle and expression of related genes in HepG2 cells. Methods : The MTT assay, cell counting assay, $[^3H]-Thymidine$ incorporation assay, flow cytometric analysis, quantitative RT-PCR and western blot assay were studied. Results : In the water extract of Loranthus parasiticus Merr., inhibition of cell proliferation and DNA synthesis in HepG2 cells was seen. These inhibitory effects were due to inhibition of G l-S transition in cell cycle. After treatment with the extract, expression of cyclin D1(G1 check point related gene) was inhibited particularly in dose-dependent and time-dependent manners. Conclusion : These results suggest that the inhibition of cell cycle progression by Loranthus parasiticus Merr. in HepG2 cell is due to suppression of cyclin D1(G1 check point related gene) mRNA expression and protein synthesis.

• Journal of Acupuncture Research
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• v.22 no.2
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• pp.155-162
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• 2005

Anemarrhenae Rhizoma (AR) has been widely used for the treatment of various diseases in Oriental medicine. To investigate whether AR possesses anti-inflammatory effects against lipopolysaccharide (LPS)-induced inflammation in the BV2 microglial cells, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide) assay. reverse transcription -polymerase chain reaction (RT-PCR), and prostaglandin E2 (PGE2) assay, and nitric oxide (NO) detection assay were performed. From the present results, AR was shown to suppress PGE2 synthesis and NO production by inhibiting the LPS-stimulated enhancement of cyclooxygenase-2 and inducible nitric oxide synthase expression in BV2 microglial cells. These results suggest that AR may offer a valuable means of therapy in the treatment of inflammatory diseases by attenuating LPS-induced PGE2 synthesis and NO production.

• Journal of Microbiology
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• v.43 no.5
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• pp.398-405
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• 2005

Polyamines such as putrescine are small, ubiquitous polycationic molecules that are required for optimal growth of eukaryotic and prokaryotic cells. These molecules have diverse effects on cell physiology and their intracellular content is regulated by de novo synthesis and uptake from the environment. The studies presented here examined the structure of a putative polyamine transporter (Pot) operon in Streptococcus pneumoniae (pneumococcus) and growth of pneumococci in medium containing putrescine substituted for choline. RT-PCR experiments demonstrated that the four genes encoding the Pot system are co-transcribed with murB, a gene involved in an intermediary step of peptidoglycan synthesis. Pneumococci grown in chemically-defined media (CDM) containing putrescine without choline enter logarithmic phase growth after 36-48 hs. However, culture density at stationary phase eventually reaches that of choline-containing medium. Cells grown in CDM-putrescine formed abnormally elongated chains in which the daughter cells failed to separate and the choline-binding protein PspA was no longer cell-associated. Experiments with CDM containing radiolabeled putrescine demonstrated that pneumococci concentrate this polyamine in cell walls. These data suggest that pneumococci can replicate without choline if putrescine is available and this polyamine may substitute for aminoalcohols in the cell wall teichoic acids.

• Korean Journal of Pharmacognosy
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• v.52 no.3
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• pp.143-148
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• 2021

This study was performed to elucidate the inhibitory effects of Alveopora japonica extract on melanin synthesis by measuring the levels of cell viability, mRNA expression, tyrosinase activity, and melanin production in the B16F10 cell line. The effects of A. japonica extract on tyrosinase-related protein 1 (TYRP1), TYRP2, tyrosinase (TYR), and microphthalmia-associated transcription factor (MITF) mRNA expression levels and melanin content were determined. Quantitative real-time RT-PCR show that A. japonica extract decrease the mRNA expression levels of TYRP1, TYRP2, TYR, and MITF in B16F10 cell line, resulting in lower levels of melanin production compared to α-MSH-treated B16F10 cells. Tyrosinase activity assays reveal that A. japonica extract decrease melanin production in B16F10 cells. These results demonstrate the whitening effects of A. japonica extract on B16F10 cells; thus, A. japonica extract is a potent ingredient for skin whitening. Further research is needed on the mechanism of action of A. japonica extract. Such research will benefit not only cosmetics, but also the health food and medical industries.

• The Journal of Internal Korean Medicine
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• v.43 no.1
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• pp.68-78
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• 2022

Objective: In this study, we investigated the protective effect of rhododendron mucronulatum extract (RME) on allergic reactions and inflammation. Methods: The effect of RME was determined using ELISA and RT-PCR in RBL-2H3 mast cells and RAW 264.7 macrophage cells. We determined cell viability, β-hexosaminidase release, and the synthesis of IL-4 and TNF-α in RBL-2H3 cells. In addition, we determined NO from RAW 264.7 and the gene expression of IL-1β, iNOS, IL-6, TNF-α, and IL-10. Results: RME inhibited β-hexosaminidase release and synthesis of IL-4 and TNF-α in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. In addition, RME inhibited the production of NO and the gene expression of IL-1β, iNOS, IL-6, and TNF-α in LPS-stimulated RAW 264.7 cells. Conclusion: From these results, we concluded that RME possesses anti-allergic activity and anti-inflammatory activity due to the inhibition of mast cells and macrophage function.

• Korean Journal of Pharmacognosy
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• v.52 no.2
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• pp.99-104
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• 2021

This study was performed to elucidated the inhibitory effects of 6,8-diprenylorobol on melanin synthesis by measuring the levels of cell viability, mRNA expression, tyrosinase activity, and melanin production in the B16F10 cell line. The effects of 6,8-diprenylorobol on tyrosinase-related protein 1 (TYRP1), TYRP2, tyrosinase (TYR), and microphthalmia-associated transcription factor (MITF) mRNA expression levels and melanin content were determined. Quantitative real-time RT-PCR shows that 6,8-diprenylorobol decreases the mRNA expression levels of TYRP1, TYRP2, TYR, and MITF in B16F10 cell line, resulting in lower levels of melanin production compared to α-MSH-treated B16F10 cells. Tyrosinase activity assays reveal that 6,8-diprenylorobol decreases melanin production in B16F10 cells. These results demonstrate the whitening effects of 6,8-diprenylorobol on B16F10 cells; thus, 6,8-diprenylorobol is a potent ingredient for skin whitening. Further research is needed on the mechanism of action of 6,8-diprenylorobol. Such research will benefit not only cosmetics, but also the health food and medical industries.

• Korean Journal of Microbiology
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• v.42 no.4
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• pp.277-285
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• 2006

The most biofilm forming bacteria in catheter, Esctherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus were isolated and identified from a patient's catheter occuring catheter-associated urinary tract infection (CA-UTI). We examined mRNA expression and its quantification of AIs synthetic genes encoding signal substance of quorum sensing from each bacterial species in order to elucidated quorum sensing mechanism. Both pure cultures for each bacterial strains and a mixed cultures with three were grown for 24 hr and 30 days. Initial densities to be able to detect mRNA expression oil single strains culture were shown at $2.4{\times}10^5$ CFU/ml, $5.4{\times}10^6$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $6.9{\times}10^4$ CFU/ml of P. aeruginosa for rhlI and lasI. Also, in mixed culture of three, initial cell densities of mRNA expression were appear to at $7.3{\times}10^5$ CFU/ml, $1.6{\times}10^7$ CFU/ml of E. coli for ygaG and S. aureus for luxS, and at $2.1{\times}10^5$ CFU/ml of P. aeruginosa for rhlI and lasI. Each AIs synthetic gene was expressed in initial cell density and the mRNA expression of the genes were detected continously during 30 days. And then, the quantification of mRNA expression level of ygaG, rhlI, last, and luxS which were related AIs synthesis was done each time point by real-time RT-PCR. Interestingly, the mRNA levels of ygaG, rhlI, lasI, and luxS from the mixed culture was higher than those from each single strain culture. In the case of E. coli ygaG, the amount of transcript from the mixed culture was at least 30 times for that from single culture. In the case of P. aeruginosa rhlI and lasI, the amount of transcript from the mixed culture was at least 40 times and 250 times for that from single strain culture. In the case of S. aureus luxS, the amount of transcript from the mixed culture was at least 5 times for that from single strain culture. And specially, the mRNA expression of rhlI and lasI of P. aeruginosa showed the highest efficency among four AIs synthetic genes.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.356-362
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• 2010

Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.