• Applied Biological Chemistry
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• v.49 no.2
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• pp.91-96
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• 2006

Antioxidant activities and biological properties such as antimutagenic and cytotoxic effect of Phellinus linteus extracts from different extraction conditions were measured against Salmonella typhimurium and human cancer cell lines. DPPH free radical scavenging activities of the extracts were higher in the solutions extracted with ethanol (17.14) and ethanol after water (17.79), respectively. In the Ames test, ethanol extract of P. linteus alone did not exhibit any mutagenicity but showed substantial inhibitory effect against mutations induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitroquinoline-1-oxide (4NQO), and benzo $({\alpha})$ pyrene $(B({\alpha})P)$. The extracts of ethanol and ethanol after water of P. linteus $(200\;{\mu}g/plate)$ had the highest inhibitory effect of 61.5 and 60.9%, respectively, on the mutagenesis on S. typhimurium TA98 strain induced by $B({\alpha})P$. Extracted solutions of ethanol and ethanol after water of P. linteus showed high antimutagenic effect against MNNG, 4NQO, Trp-P-1 and $B({\alpha})P$, causing mutations in S. typhimurium TA100 strain. The anticancer effects of P. linteus extracts were investigated against human fibrosarcoma HT-29 and human hepatocellular carcinoma HepG2. The treatment of 0.5 mg/ml of ethanol, ethanol after water and water extracts of P. linteus had the highest cytotoxicity of 59, 57, 54%, respectively against HT-27 cell line, whereas low cytotoxicity effects were observed against HepG2 cell line in the range of $10{\sim}30%$. The ethanol and water extracts of P. linteus also showed the nitrate scavenging ability at different pHs. The ethanol extract showed higher nitrate-scavenging ability compared to water extract of P. linteus.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.215-225
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• 2005

Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Oligonucleotide microarray approach were employed to screen the differential expression genes. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CTF-HAS(0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cytotoxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of CTF-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome U133 Plus 2.0., Affimatrix Co.). ResuIts : It has no cytotoxic effects on HepG2 cells in all concentrations (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$). More than twofold up-regulated genes were 19 genes. The number of more than twofold down-regulated genes was 13. Discussion : This study showed the screening of CTF-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

• Korean Journal of Medicinal Crop Science
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• v.11 no.1
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• pp.53-61
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• 2003

Cirsium japonicum var. ussuriense were extracted with methanol and then fractionated with nhexane, EtOAc and BuOH to get active fractions. And their antioxidant and antimicrobial activities in each fraction were determined. Ethyl acetate and butanol fraction of Cirsium japonicum var. ussuriense showed strong antioxidant activities, but hexane fraction did not show any activities. But in the antimicrobial test, Ethyl acetate fraction showed strong antimicrobial activities except to Aspergillus awamori, Asperigillus niger. Especially, Ethyl acetate fraction showed the strongest activities against Bacillus subtilis. And aqueous fraction showed the strongest activities against Cladosporium herbarum, Hypocrea nigricans. This study was performed to determine the antimutagenic and cytotoxic effect of Cirsium japonicum var. ussuriense methanol extract on Salmonella typhimurium TA98, TA100 and cancer cell lines using Ames test and cytotoxicity assay, respectively. Cancer cell lines include human lung carcinoma(A549), human breast adenocarcinoma(MCF-7) and human hepatocellular carcinoma (Hep3B). Futher fractionations with hexane, ethyl acetate, butanol and water from methanol extract of Cirsium japonicum var. ussuriense were performed to obtain effective fraction, methanol extract showed 60.14% inhibition effect on the mutagenesis induced by MNNG against TA100, while 77% and 72.5% inhibition was observed on the mutagenesis induced by 4NQO against TA98 and TA100, respectively. and methanol extract showed 82.25% and 73.7% inhibitory effect on the mutagenesis induced by Trp-P-1 against TA98 and TA100, respectively. methanol extract showed the strongest effect against A549, MCF-7 and Hep3B at the same concentration compared to those of other fration.

• Journal of Food Hygiene and Safety
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• v.38 no.2
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• pp.69-78
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• 2023

This study aimed to investigate the protective effect of enzymatically modified stevia (EMS) on C2C12 cell-based model of dexamethasone (DEX)-induced muscle atrophy to provide baseline data for utilizing EMS in functional health products. C2C12 cells with DEX-induced muscle atrophy were treated with EMS (10, 50, and 100 ㎍/mL) for 24 h. C2C12 cells were treated with EMS and DEX to test their effects on cell viability and myotube formation (myotube diameter and fusion index), and analyze the expression of muscle strengthening or degrading protein markers. Schisandra chinensis Extract, a common functional ingredient, was used as a positive control. EMS did not show any cytotoxic effect at all treatment concentrations. Moreover, it exerted protective effects on C2C12 cell-based model of DEX-induced muscle atrophy at all concentrations. In addition, the positive effect of EMS on myotube formation was confirmed based on the measurement and comparison of the fusion index and myotube diameter when compared with myotubes treated with DEX alone. EMS treatment reduced the expression of muscle cell degradation-related proteins Fbx32 and MuRF1, and increased the expression of muscle strengthening and synthesis related proteins SIRT1 and pAkt/Akt. Thus, EMS is a potential ingredient for developing functional health foods and should be further evaluated in preclinical models.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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• v.12 no.1
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• pp.46-54
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• 2001

Background and Objectives : Nitric oxide(NO) is a short-lived molecule with messenger and cytotoxic functions in nervous, cardiovascular, and immune systems. Among the three distinct NOS isoforms, the neuronal isoform is expressed in small, discrete neuronal populations of CNS and PNS. Axonal injury in adult animals results in a dramatic NOS up-regulation in many types of central and peripheral neurons which normally lack the enzyme or express it only at very low levels. In previous study, we confirmed the efficacy of PEMS on the early functional recovery in rats with surgically transected and reanastomosed recurrent laryngeal nerve. Therefore, after we obtained functionally recovered rats using PEMS in this study, we studied to evaluate the expression of nNOS through the analysis of the difference between functional recovery group and non-recovery group in the recurrent laryngeal nerve. Materials and Method : Using 84 healthy male Sprague-Dawley rats, transections and primary anastomosis were performed on their left recurrent laryngeal nerves. Rats were then randomly assigned to 2 groups. The rats in group A(n=42) received PEMS by placing them in custom cages equipped with Helm-holz coils(3 hr/day, 5 days/wk, for 12 wk). The rats in group B(n=42) were handled the same way as the group A, except that they did not receive PEMS. Laryngovideoendoscopy was performed before and after surgery and followed up weekly. Laryngeal EMG was obtained in both PCA and TA muscles. Immunohistochemisty staining using monoclonal anti-neuronal nitric oxide synthase(nNOS) antibody was performed to detect nNOS in recurrent laryngeal nerve and nodose ganglion. Results : 20 rats(63%) in group A and 5 rats(17%) in the group B showed recovery of vocal fo1d motion. The number of NOS-positive cells was increased in functionally-recovered rats. NOS-staining intensity was reduced 12 weeks after nerve injury. The difference between PEMS group and non-stimulated group was not found. Conclusion : This study shows that nNOS may exert a beneficial effect on nerve regeneration and functional repair.

• The Korea Journal of Herbology
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• v.23 no.2
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• pp.179-192
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• 2008

Objectives : Draconis Resina (DR) has been used as a traditional Korean herbal medicine since ancient times, and today it is used as a medication for wounds, tumors, diarrhea, rheumatism, in the itching of insect bites and with other conditions in the folk medicine. The aim of this study was to determine whether fractionated extracts of DR inhibit free radical generation, intracellular oxidation, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, Methods : DR extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of DR onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced H202, NO, PGE2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 formation in macrophages. Conclusions : Our results indicate that dichloromethane and ethyl acetate extracts of DR have potential as an agent of chronic inflammatory diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.8
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• pp.1128-1136
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• 2015

This study investigated the anti-inflammatory effects of ethanol extract from Grateloupia elliptica Holmes (GEHEE) on the lipopolysaccharide-induced inflammatory response. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There were no cytotoxic effects on proliferation of macrophages treated with GEHEE compared to the control. GEHEE remarkably suppressed NO and pro-inflammatory cytokines (interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$) production and reduced expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) proteins in a dose-dependent manner. GEHEE also significantly reduced activation of mitogen-activated protein kinases (MAPKs). The formation of edema in mouse ears was reduced at the highest dose compared to the control. GEHEE also reduced dermal thickness and mast cell numbers based on histological analysis. These results suggest that GEHEE exerts significant anti-inflammatory activity via inhibition of $NF-{\kappa}B$ and MAPKs activation and may be a potential anti-inflammatory therapeutic material.

• Journal of Korean Traditional Oncology
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• v.16 no.2
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• pp.25-34
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• 2011

Objectives : Citrus is the fruit that is readily available around us. Therefore, we investigated the anti-inflammatory effects of fraction isolated from the Citrus hassaku pericarp in RAW264.7 macrophage cells. Methods : The effects of fraction from Citrus hassaku pericarp on cell viability on RAW264.7 cells were measured by the MTT assay. The mRNA levels of iNOS and COX-2, its protein level by fraction of Citrus hassaku pericarp treatment in RAW264.7 macrophage cells were investigated by RT-PCR and immunoblots. Nitrite accumulation in the culture was measured colorimetrically by the Griess reaction using a Griess reagent. The amount of IL-6 and TNF-${\alpha}$ production was determined using an enzyme-linked immunosorbent assay (ELISA) kit. Results : The results indicated that the fraction of Citrus hassaku pericarp concentration highly suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) and IL-6 productions without a cytotoxic effect on RAW264.7 cells. fraction of Citrus hassaku pericarp inhibited the expressions of LPS-induced iNOS and COX-2 protein and their mRNA in a dose-dependent manner. Particularly, fraction of Citrus hassaku pericarp suppressed the level of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity, which was linked with the suppression of LPS-induced phosphorylation of p65 at serine 276 and p65 translocation into nuclei, but not MAPK signaling. In addition, treatment with fraction of Citrus hassaku pericarp inhibited the production of IL-6 and TNF-${\alpha}$ in LPS-stimulated RAW264.7 cells. Conclusion : Our results indicate that fraction of Citrus hassaku pericarp potentially inhibits the biomarkers related to inflammation through the blocking of NF-${\kappa}B$ p65 activation, and it may be a potential therapeutic candidate for the treatment of inflammatory diseases.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.3
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• pp.247-254
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• 2012

Ramalin (${\gamma}$-glutamyl-N'-(2-hydroxyphenyl)hydrazide) isolated from the Antarctic lichen Ramalina terebrata has been shown to have strong antioxidant activities in the previous study. To investigate additional activities of ramalin, we studied the effects of ramalin on melanogenesis in melan-a cells, a non-tumorigenic melanocyte cell line. At a non-cytotoxic concentration, ramalin dramatically decreased melanin synthesis in melan-a cells in a dose-dependent manner, which was more potent than arbutin, a well-known tyrosinase inhibitor. Ramalin inhibited cell-free tyrosinase activity directly and intracellular tyrosinase activity as well. Its inhibitory mechanisms on melanin production were further assessed, and we found that ramalin significantly decreased the protein levels of melanogenic enzymes such as tyrosinase, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2). However, the mRNA levels of these enzymes were not altered. In a clinical study, application of 0.2 % ramalin on human skin significantly improved the degree of skin brightness after 3 weeks. In conclusion, ramalin has strong anti-melanogenic activity that is exerted both by the direct inhibition of tyrosinase activity and by down-regulation of melanogenic proteins. Furthermore, ramalin showed skin brightening effect in a clinical study. Collectively, these results suggest that ramalin may be a useful inhibitor for melanogenesis in skin.

• Journal of Life Science
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• v.28 no.7
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• pp.802-810
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• 2018

Aging is accompanied by changes in the body, such as graying hair, wrinkles, and black spots composed of lipid peroxides and proteins. Melanin is a polymer substance produced by an oxidation polymerization reaction from tyrosine, and it determines the color of hair and skin. It has been reported that melanin is synthesized by melanocyte, and its excessive production by reactive oxygen species is associated with aging. The purpose of this study was to determine the direct effects of Musa paradisiaca peel ethanolic extract (MPEE) on antioxidative activity and melanin synthesis. It was observed that the antioxidant activity of MPEE was similar to that of vitamin C, a positive control, in both DPPH radical scavenging assay and reducing power assay. In order to examine cytotoxicity prior to cell experimentation, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed for B16F1 cells. MPEE was not cytotoxic at $32{\mu}g/ml$ or less. In addition, MPEE increased melanin synthesis in live cells in addition to tyrosinase activity and melanin synthesis in dihydroxyphenylalanine (DOPA)-oxidation assay in vitro. Moreover, MPEE increased melanin synthesis in cells aged by pretreatment with $H_2O_2$. The expression levels of tyrosinase-related protein (TRP)-1, TRP-2, and superoxide dismutase (SOD)-2 by western blot analysis were increased in the presence of MPEE. These results suggest that MPEE could promote the melanin synthesis as an antioxidative substance.