• Food Science and Preservation
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• v.18 no.6
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• pp.938-945
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• 2011

Epimedium koreanum Nakai is a wild herb commonly consumed in South Korea due to its beneficial health effects. In the present study, the antimutagenic and immunoactivities of extracts from E. koreanum Nakai containing different icariin quantities were investigated for food use. In the Ames test, both the water and ethanol extracts were found not to have a mutagenic effect on Salmonella typhimurium TA98 and TA100, respectively. The E. koreanum Nakai extracts showed over 80 and 90% antimutagenic effects on benzo(${\alpha}$)pyrene (B(a)P) in S. typhimurium TA98 and TA100, respectively. Moreover, all the extracts showed over 70% antimutagenicity on S. typhimurium TA98 and TA100 against 4-nitroquinoline-1-oxide (4NQO). The E. koreanum Nakai extract with ethanol showed strong antimutagenic activity, higher than that of the water extract and the sequenced KE9412, KE9408, and KE9405. In the immunomodulating activity test, the effect of E. koreanum Nakai on the B (Rhamos) and T (Jukat) cells were investigated. The immunoactivity results showed that the growth and viability of the B and T cells increased and were activated more in KE9405 (1.8 times), KE9408 (1.6 times), and KE9412 (1.32 times) in the water extracts, and least in KE9412 (1.74 times), KE9408 (1.52 times), and KE9405 (1.4 times) in the ethanol extracts. In the case of both the water and ethanol extracts ($500{\mu}g/mL$) from E. koreanum Nakai, the highest cell number of the human B (Rhamos) and T (Jukat) cells was observed on day 4 in KE9405 and KE9412, and on day 5 in KE9408. Based on the obtained results, the development of E. koreanum Nakai as a food material is recommended.

• Food Science and Preservation
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• v.21 no.6
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• pp.878-884
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• 2014

Increased antimutagenic and in vitro anticancer effects were observed by the fermentation process during Kochujang manufacturing. In order to confirm the increased functionality, wheat grain, first fermented wheat grains (FFWG), second fermented wheat grains (SFWG), final fermented wheat grains (FiFWG), red pepper powder (RPP), and commercial Kochujang (CK) were prepared. Kochujang manufactured with final fermented wheat grains and red pepper powder were further fermented for 15 days and 30 days. The antimutagenic effects were determined by counting the number of revertants in Salmonella Typhimurium TA100 against N-methyl-N'-nitro-nitrosoguanidine (MNNG, 1.0mg/mL). The final fermented wheat grains (52% inhibition) showed higher antimutagenic effects than the wheat grain (34%), and the commercial Kochujang showed the highest antimutagenic effects (55%). We tested the inhibitory effect on the growth of HT-29 human colon carcinoma cells and AGS human gastric adenocarcinoma cells by using MTT (3-(4,5-dimethyl-thiazol)-2,5-diphenyltetrazolium bromide) assay. The results showed that increased fermentation process continually increased the growth inhibitory effect on both cancer cells. The further fermentation for 15 days of the Kochujang product also increased inhibitory growth of the AGS cancer cells. In conclusion, the methanol extract from fermented wheat grains and commercial Kochujang showed sequentially increased antimutagenic and in vitro anticancer activity, and thus the final commercial Kochujang revealed the highest effect.

• Journal of Pharmacopuncture
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• v.20 no.2
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• pp.127-131
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• 2017

Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

• The Journal of Internal Korean Medicine
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• v.32 no.2
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• pp.153-169
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• 2011

Objectives : This study was performed to investigate the anti-fibrogenic effect and the effect on cell growth and apoptosis in YJCGT, YJCGT YSO and YJCGT YSCO on thioacetamide-induced rat liver tissue and the immortalized human hepatic cell line LX2. Materials and Methods : LX2 cells were treated with various concentrations (0, 50, 150, 300 ug/ml) of YJCGT, Y+YSO, and Y+YSCO extract for 24, 48 and 72 hours. After the treatment, cell viability was measured by using MTT assay. Caspase inhibitor assay, and cell viability were determined by a colorimetric assay with PMS/MTS solution. Rat liver fibrosis was induced by intraperitoneal thioacetamide injection 150 mg/kg 3 times a week for 5 weeks. After the treatment, body weight, liver & spleen weights, liver function test, the complete blood cell count and the change of portal pressure were studied. After YJCGT, Y+YSO, and Y+YSCO treatment, percentages of collagen in thioacetamide-induced rat liver tissue were measured. Results : The viability of the LX2 cell decreased in a dose- and time-dependent manner. Exposure of LX2 cells to YJCGT, YJCGT+YSO and YJCGT+YSCO induced caspase-3 activation, but co-treatment of YJCGT, YJCGT+YSO and YJCGT+YSCO with the pan-caspase inhibitor Z-VAD-FMK, and the caspase-3 inhibitor Z-DEVE-FMK, blocked apoptosis. There was no difference in rat body weight between the thioacetamide only group and the YJCGT, YJCGT+YSO and YJCGT+YSCO groups. In the YJCGT, YJCGT YSO and YJCGT YSCO groups, the serum level of GPT significantly went down compared with the thioacetamide only group. In the YJCGT, Y+YSO, Y+YSCO groups, white blood cell elevated by thioacetamide injection decreased but RBC, Hgb, and Hct increased. In the Y+YSO group, the portal pressure elevated by thioacetamide injection significantly decreased. In the histological finding, thioacetamide injections caused severe fibrosis, but YJCGT, Y+YSO, and Y+YSCO treatment significantly reduced the amounts of hepatic collagens. Conclusions : YJCGT, Y+YSO, and Y+YSCO inhibit the growth of LX2 cells by inducing apoptosis through caspase activity. YJCGT, Y+YSO, and Y+YSCO have beneficial effects on the treatment of cirrhotic patients as well as patients with chronic hepatitis.

• Journal of Preventive Medicine and Public Health
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• v.39 no.3
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• pp.199-204
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• 2006

Objectives: Mercury is a hazardous organ-specific environmental contaminant. It exists in a wide variety of physical and chemical states, each of which has unique characteristics for the target organ specificity. Exposure to mercury vapor and to organic mercury compounds specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds. Methods: In this study, mercury chloride $(HgCl_2)$ was studied in a renal derived cell system, i.e., the tubular epithelial Madin-Darby canine kidney (MDCK) cell line, which has specific sensitivity to the toxic effect of mercury. MDCK cells were cultured for 6-24 hr in vitro in various concentrations (0.1-100 M) of $HgCl_2$, and the markers of apoptosis or cell death were assayed, including DNA fragmentation, caspase-3 activity andwestern blotting of cytochrome c. The influence of the metal on cell proliferation and viability were evaluated by the conventional MTT test. Results: The cell viability was decreased in a time and concentration dependent fashion: decreases were noted at 6, 12 and 24 hr after $HgCl_2$, exposure. The increases of DNA fragmentation were also observed in the concentrations from 0.1 to 10 M of $HgCl_2$ at 6 hr after exposure. However, we could not observe DNA fragmentation in the concentrations more than 25 M because the cells rapidly proceeded to necrotic cell death. The activation of caspase-3 was also observed at 6 hr exposure in the $HgCl_2$ concentrations from 0.1 to 10 M. The release of cytochrome c from the mitochondria into the cytosol, which is an initiator of the activation of the caspase cascade, was also observed in the $HgCl_2-treated$ MDCK cells. Conclusions: These results suggest that the activation of caspase-3 was involved in $HgCl_2-induced$ apoptosis. The release of cytochrome c from the mitochondria into the cytosol was also observed in the $HgCl_2-treated$ MDCK cells. These findings indicate that in MDCK cells, $HgCl_2$ is a potent inducer of apoptosis via cytochrome c release from the mitochondria.

• Journal of Sericultural and Entomological Science
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• v.50 no.1
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• pp.20-26
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• 2012

In this study, silk gland powder of silkworm were investigated to see the possibility for baby powder cosmetics materials. To test possibility as a baby powder cosmetics, total content rate of amino acids, DPPH free radical scavenging assay, MTT assay, and clinical trial were done. According to the result of the analysis of the amino acids of silk gland powder, serin (26.77%) content was the highest and asparatic acid (15.47%), and glycine (9.62%) were followed. DPPH free radical scavenging activity of silk gland powder was lower than vitamin C by 82.3% and 97%, respectively, which is relatively good. Moisture effect were increased in silk gland powder compared to control cosmetics by 50%. Also, silk gland powder was classified as a practically non-irritating material based on the score 0.05 of primary irritation index. Thus, these results suggest that silk gland powder of silkworm may have beneficial properties as a material for baby powder cosmetics.

• Radiation Oncology Journal
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• v.12 no.2
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• pp.129-141
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• 1994

Intracellular ions which have a major role in cellular function have been reported to affect repair of radiation damage. Recently it has been reported that ouabain sensitizes A549 tumor cellls but not CCL-120 normal cells to radiation. Ouabain inhibits the $Na^+-K^+$-pump rapidly thus it increases intracellular Na concentration, Vanadate which is distributed extensively in almost all living organisms is known to be a $Na^+-K^+$-ATPase inhibitors, This study was performed to see any change in radiosensitivity of tumor cell by vanadate and any role of $Na^+-K^+$ATPase in radiosensitization. Experiments have been carried out by pretreatment with vanadate in human cell line(A549, JMG) and mouse cell line(L1210, spleen). For the cell survival MTT assay was performed for A549 and JMC cells and frypan blue dye exclusion test for L120, and spleen cells. Measurements of $Na^+-K^+$-ATPase activity in control, vanadate treated cell, radiation treated cell (9 Gy for A549 and JMG, 2 Gy for L1201, spleen), and combined $10^{-6}M$ vanadate and radiation treated cells were done. The results were summerized as fellows. 1. L1210 cell was most radiosensitive, and spleen cell and JMG cell were intermediate, and A549 cell was least radiosensitive. 2. Mininum or no cytotoxicity was seen with vanadate below concentration of $10^{-6}M$. 3. In A549 cells there was a little change in radiosensitivity with treatment of vanadate. However radiation sensitization was shown in low dose level of radiation i. e. 2- Gy. In JMG cells no change in radiosensitivity was noted. Both L1210 and spleen cell had radiosensitization but change was greater in tumor cell. 4. $Na^+-K^+$-ATPase activity was inhibited significantly in tumor cell by treatment of vanadate. 5. Radiaiton itself inhibited $Na^+-K^+$-ATPase activity of tumor cell with high $Na^+-K^+$-ATPase concention. Increase in radiosensitivity by vanadate was closely associated with orginal $Na^+-K^+$-ATPase contents. From the above results vanadate had little cytotoxicity and it sensitized tumor cells to radiation. Inhibitory effect of vanadate on $Na^+-K^+$-ATPase activity might be one of the contributing factors for radiosensitization to tumor cells which has greater enzyme activity than that of normal cell. It was suggested vanadate could be used as a potential radiosensitizer for tumor cells.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.1
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• pp.489-494
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• 2014

Background: Psychiatric patients appear to be at lower risk of cancer. Some antipsychotic drugs might have inhibitory effects on tumor growth, including penfluridol, a strong agent. To test this, we conducted a study to determine whether penfluridol exerts cytotoxic effects on tumor cells and, if so, to explore its anti-tumor mechanisms. Methods: Growth inhibition of mouse cancer cell lines by penfluridol was determined using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cytotoxic activity was determined by clonogenic cell survival and trypan blue assays. Animal tumor models of these cancer cells were established and to evaluate penfluridol for its anti-tumor efficacy in vivo. Unesterified cholesterol in cancer cells was examined by filipin staining. Serum total cholesterol and tumor total cholesterol were detected using the cholesterol oxidase/p-aminophenazone (CHOD-PAP) method. Results: Penfluridol inhibited the proliferation of B16 melanoma (B16/F10), LL/2 lung carcinoma (LL/2), CT26 colon carcinoma (CT26) and 4T1 breast cancer (4T1) cells in vitro. In vivo penfluridol was particularly effective at inhibiting LL/2 lung tumor growth, and obviously prolonged the survival time of mice bearing LL/2 lung tumors implanted subcutaneously. Accumulated unesterified cholesterol was found in all of the cancer cells treated with penfluridol, and this effect was most evident in LL/2, 4T1 and CT26 cells. No significant difference in serum cholesterol levels was found between the normal saline-treated mice and the penfluridol-treated mice. However, a dose-dependent decrease of total cholesterol in tumor tissues was observed in penfluridol-treated mice, which was most evident in B16/F10-, LL/2-, and 4T1-tumor-bearing mice. Conclusion: Our results suggested that penfluridol is not only cytotoxic to cancer cells in vitro but can also inhibit tumor growth in vivo. Dysregulation of cholesterol homeostasis by penfluridol may be involved in its anti-tumor mechanisms.

• Korean Journal of Oriental Medicine
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• v.18 no.3
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• pp.103-110
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• 2012

Objectives : Fire needling has been applied as the treatment for various diseases and been getting much attention from Oriental medicine due to its excellent effectiveness as the results of clinical studies have reported. However, the research findings on the safety of treatment method, materials for the Fire needling needle materials and the possibility of burn injury during the procedure are still insufficient. Methods : A thermo imaging camera was used to confirm the temperature distribution on acupuncture needle and the treatment area during the fire needling therapy. Then the degree of thermal injury was observed by H&E stain and TUNEL assay. In addition, in order to assess the safety of acupuncture materials, we conducted MTT assay using a L6 cell line. Results : The average temperature of the skin surface was observed at $47{\sim}51^{\circ}C$ after classic fire needling and $30^{\circ}C$ after warming fire needling. Warming fire needling therapy does not induce a burn on the tissue and a third degree burn was observed locally in the muscle and skin layers after classic fire needling treatment. This confirms that hwa-acupuncture therapies do not cause major burns. According to the safety assessment test result, no cytotoxicity was detected in the warming fire needling materials. This confirms the safety of the acupuncture materials Conclusions : Various research results on the biological safety of fire needling. Since fire needling therapy induces a burn locally without leaving any scar, and as other results indicate, it is considered a safe treatment method.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.31 no.3
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• pp.39-49
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• 2018

Objectives : This study is designed to clarify whitening, anti-inflammatory effect of fractions extracted from the mixture of Phaseolus radiatus L. and ethanol. Methods : In this experiment, we were intended to reveal whitening, anti-inflammatory effect of fractions extracted from the mixture of Phaseolus radiatus L. and ethanol. The whitening activity was confirmed by UV blocking activity, tyrosinase inhibiting activity, and melanin formation inhibiting activity. Anti-inflammatory activity is confirmed by measurement of cytotoxicity level by MTT assay and measurement of Cytokine expression, which is the main mediator of inflammation reaction. Results : As a results, overall activity was high in the ethyl acetate fraction. Tyrosinase inhibitory activity was less than 20% at all concentrations, but the activity to inhibit melanin self-production was higher than that of ethyl acetate fraction at $32.19{\pm}2.79%$ at $100{\mu}g/m{\ell}$. And ethyl acetate fraction had a relatively high UV blocking activity. In the anti-inflammatory test, the concentration-dependent activity was shown, and the chloroform and ethyl acetate fractions showed significant NO production inhibitory activity. Cytokine expression was superior to that of the final stage of B cell differentiation, and cell viability was over 80% except for the chloroform fraction at the concentration of $200{\mu}g/m{\ell}$. Conclusions : The results of this experiment confirmed the whitening effect and anti-inflammatory effect of Phaseolus radiatus L.'s extracts and fractions and report the possibility of application as external medicine.