• Journal of Haehwa Medicine
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• v.7 no.1
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• pp.939-955
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• 1998

Achyranthis Bidentatae Radix(ABR) is important prescriptions that have been used in oriental medicine for stomatitis and wound healing. The study was done to evaluate the inhibitory effects of cytotoxicity, formation of superoxide on the macrophage and neutrophil, prostaglandins($PGE_2$), interleukins($IL-1{\beta}$), collagenase activity and synthesis of collagen and DNA. The results were obtained as follows: 1. ABR was not showed the proliferation difference of human fibroblast and monocyte in 0.01% and 0.001% concentrations to be experimented and in result, it was concluded that they have no cytotoxicity but showed cytotoxicity in 0.1% concentrations. 2. ABR inhibited the formation of superoxide to 48% at the concentration of 0.001% in the mouse monocyte. 3. ABR inhibited the formation of superoxide to 40% at 0.001%, 58% at 0.0001% as compared with control in the human monocyte. 4. ABR inhibited the formation of superoxide to 58% at 0.0001%, 40% at 0.001% in the human neutrophil. 5. ABR was not showed the proliferation difference of human monocyte in all concentrations to be experimented and in result, it was concluded that they inhibited the formation of prostaglandins($PGE_2$) in the human monocyte stimulated with E. coli. 6. ABR showed the all concentration of inhibiting the production of inter1eukins($IL-1{\beta}$) in the human monocyte stimulated with E. coli. 7. ABR didn't influence on collagen synthesis and total protein in fibroblasts. 8. ABR inhibited the collagenase activity to 84% at 0.1%, 69% at 0.2%, 76% at 0.5%, 91% at 0.001% respectively.

• Journal of Haehwa Medicine
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• v.7 no.1
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• pp.837-852
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• 1998

Ulmus parvifolia(UP) is important prescriptions that have been used in oriental medicine for stomatitis and wound healing. The study was done to evaluate the inhibitory effects of cytotoxicity, formation of superoxide on the macrophage and neutrophil, prostaglandins($PGE_2$), interleukins($IL-1{\beta}$), collagenase activity and synthesis of collagen and DNA. The results were obtained as follows: 1. UP was not showed the proliferation difference of human fibroblast and monocyte in all concentrations to be experimented and in result, it was concluded that they have no cytotoxicity. 2. UP inhibited the formation of superoxide to 22% at 0.01%, 52% at 0.001% in the mouse monocyte. 3. UP inhibited the formation of superoxide to 6% at the concentration of 0.001% as compared with control in the human monocyte. 4. UP was not showed the proliferation difference of human neutrophil in all concentrations to be experimented and in result, it was concluded that they inhibited the formation of superoxide. 5. UP was not showed the proliferation difference of human monocyte in all concentrations to be experimented and in result, it was concluded that they inhibited the formation of prostaglandins($PGE_2$) in the human monocyte stimulated with E. coli. 6. UP was showed the all concentration of inhibiting the production of interleukins($IL-1{\beta}$) to slight in the human monocyte stimulated with E. coli. 7. UP influence on collagen synthesis and total protein in fibroblasts to at the slight of 0.05%, specially to excellent to 0.2%. 8. UP inhibited the collagenase activity to 20% at 0.1%, 31% at 0.2%, 45% at 0.5%, 24% at 0.01% respectively.

• Food Science and Biotechnology
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• v.16 no.1
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• pp.163-166
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• 2007

The cytotoxic effects of partially purified substances from Bacillus polylfermenticus SCD toward a variety tumor cell lines were studied. Cytotoxic activity was determined with regard to the A549 (human lung carcinoma), AGS (human stomach adenocarcinoma), DLD-1 (human colon adenocarcinoma), HEC-1-B (human uterus adenocarcinoma), SW-156 (human kidney carcinoma), and NIH/3T3 (murine normal fibroblast) cell lines using the MTT assay. Cytotoxic substances were partially purified through Diaion HP-20 columns and extracted with methanol or other organic solvents (n-hexane, chloroform, ethylacetate, and butanol). B. polyfermenticus SCD supernatant showed up to 60% inhibition of cell viability fer all five human cancer cell lines tested. When treated with 10 mg/mL of n-hexane, chloroform, ethylacetate, and butanol extract, HEC-1-B cells showed a 25,62,35, and 63% rate of inhibition respectively, and AGS cells showed a 72, 61, 44, and 67% rate of inhibition, respectively. At a concentration of 10 mg/mL, 100% methanol Diaion HP-20 extracts showed inhibition rates of 97.0% toward A-549 cells, 98.1% toward AGS cells, 81.6% toward DLD-1 cells, 83.5% toward HEC-1-B cells, and 92.7% toward SW-156 cells. These results indicate that partially purified fractions from B. polyfermenticus SCD have the potential to inhibit not only colon cancer cells, but also lung, stomach uterus, and kidney cancer cells. Further studies are needed to characterize the cytotoxic substances released in B. polyfermenticus SCD cultures.

• Journal of Periodontal and Implant Science
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• v.28 no.4
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• pp.703-721
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• 1998

In order to observe the effects of Nicotine and NNK on cultured human gingival fibroblast, several factors were examined including mutagenicity, the number of cells attached culture plate surface through MTT test, the abundance of collagen & collagenase in mRNA level and collagenolytic activity in extracellular matrix. The results were as follows; 1. Regardless of the co-existence of S9, Nicotine did not show the mutagenicity by itself and NNK by itself showd the same result; However, dose related mutagenicity was shown in NNK with S9. 2. The number of fibroblasts attached cultured plate surface was measured by MTT procedure. The number of cells in Non-smokers increased at all time periods as compared to those of smoker. 3. Non-smoker's fibroblast treated by NNK or Nicotine was dosedependently dosedependently decreased in the number of cells when compared to untreated control. In higher dose, Nicotine showed the cellular toxicity, but NNK did not. 4. No change in the abundance of mRNA for pro${\alpha}1$ and pro${\alpha}2$ was shown in Nicotine treated group but in gingival fibroblasts following treatment with NNK, the abundance of mRNA for pro${\alpha}1$, but not pro${\alpha}2$ collagen was decreased. 5. The abundance of mRNA for collagenase was decreased when NNK was treated but no change occurred in Nicotine treated group. 6. The effect of NNK and Nicotine in collagenolytic activity showed that, collagenase activity exclusively react to type I collagen, was increased in both group, but gelatinase exclusively react to type IV collagen was not influenced at all. Collagenase activity of smoker's fibroblast was also increased as much as Nicotine and NNK group. The findings suggest that both of Nicotine and NNK lead gingival fibroblast to decrease in the abundance of collagen. And it seems to be that Nicotine and NNK have independent pathway toward the gingival fibroblast.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.19-27
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• 2004

Objective: This study was to examine the in vitro neural cell differentiation patterns of human embryonic stem (hES) cells following treatment of various neurotrophic factors [basic fibroblast growth factor (bFGF), retinoic acid (RA), brain derived neurotrophic factor (BDNF) and transforming growth factor (TGF)-$\alpha$], particulary in dopaminergic neuron formation. Methods: The hES cells were induced to differentiate by bFGF and RA. Group I) In bFGF induction method, embryoid bodies (EBs, for 4 days) derived from hES were plated onto gelatin dish, selected for 8 days in ITSFn medium and expanded at the presence of bFGF (10 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14 and 21 days. Group II) For RA induction, EBs were exposed of RA ($10^{-6}M$) for 4 days and allowed to differentiate in N2 medium for 7, 14 and 21 days. Group III) To examine the effects of additional neurotrophic factors, bFGF or RA induced cells were exposed to either BDNF (10 ng/ml) or TGF-$\alpha$ (10 ng/ml) during the 21 days of final differentiation. Neuron differentiation and dopamine secretion were examined by indirect immunocytochemistry and HPLC, respectively. Results: The bFGF or RA treated hES cells were resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with BDNF or TGF-$\alpha$ during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression of a dopaminergic neuron marker, compared to control (p<0.05). In contrast, no effect was observed on the rate of mature neuron (NF-200) or glutamic acid decarboxylase-positive neurons. Immunocytochemistry and HPLC analyses revealed the higher levels of TH expression (20.3%) and dopamine secretion (265.5 $\pm$ 62.8 pmol/mg) in bFGF and TGF-sequentially treated hES cells than those in $\alpha$ RA or BDNF treated hES cells. Conclusion: These results indicate that the generation of dopamine secretory neurons from in vitro differentiated hES cells can be improved by TGF-$\alpha$ addition in the bFGF induction protocol.

• The Korean Journal of Food And Nutrition
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• v.30 no.2
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• pp.211-217
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• 2017

In the present study, we investigated the protective effect of various grain methanolic extracts against UVB-induced photo-aging in human skin fibroblasts. Various grain methanolic extracts were evaluated for their antioxidant compounds and activities. 2,2-Ddiphenyl-1-picryhydrazyl radical (DPPH) and ABTS 2,2-azino-bris-(3-ethylbenzoth iazoline-6-sulphonic acid) radical cation scavenging activities have been used to measure the relative antioxidant activities of extracts from grains. The content of total polyphenolics in the extracts were evaluated using spectrophotometric methods. Human skin fibroblast (Hs68) cells were pretreated with various grain methanolic extracts ($25{\mu}g/mL$). Skin toxicity was simulated by exposing the cells to UVB ($30mJ/cm^2$) irradiation. In response to the UVB-irradiation, an increased amount of matrix metallo-proteinases (MMPs) release was observed, whereas pretreatment of various grain methanolic extracts significantly inhibited the production of MMP-1 in Hs68 cells. We also found that pretreatment of the extracts significantly decreased UVB-induced reactive oxygen species and significantly increased total collagen content in Hs68 cells. These results provide that grains could be regarded as a potential ingredient in natural cosmetics, used for UVB protection.

• Journal of Food Hygiene and Safety
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• v.32 no.3
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• pp.243-248
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• 2017

Intracellular and extracellular oxidative stress initiated by reactive oxygen species (ROS) causes skin aging, which is characterized by wrinkles and atypical pigmentation. Use of antioxidant is an effective approach to prevent symptoms related to ROS-induced aging of the skin. Therefore, the antioxidant and anti-aging effect of Castanea crenata Siebold & Zucc. extracts (LCE) was investigated in this study. The LCE markedly reduced the hydrogen peroxide-induced cell damage, intracellular ROS, and oxidative stress-induced senescence in human dermal fibroblasts (HDFs). These results indicate that LCE might have beneficial effects on oxidative stress-induced damage and thus reduce skin aging.

• Nutrition Research and Practice
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• v.1 no.2
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• pp.105-112
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• 2007

Ascorbic acid has been reported to extend replicative life span of human embryonic fibroblast (HEF). Since the detailed molecular mechanism of this phenomenon has not been investigated, we attempted to elucidate. Continuous treatment of HEF cells with ascorbic acid at ($200{\mu}M$) from 40 population doubling (PD) increased maximum PD numbers by 18% and lowered $SA-{\beta}-gal$ positive staining, an aging marker, by 2.3 folds, indicating that ascorbic acid extends replicative life span of HEF cells. Ascorbic acid treatment lowered DCFH by about 7 folds and Rho123 by about 70%, suggesting that ascorbic acid dramatically decreased ROS formation. Ascorbic acid also increased aconitase activity, a marker of mitochondrial aging, by 41%, indicating that ascorbic acid treatment restores age-related decline of mitochondrial function. Cell cycle analysis by flow cytometry revealed that ascorbic acid treatment decreased G1 population up to 12%. Further western blot analysis showed that ascorbic acid treatment decreased levels of p53, phospho-p53 at ser 15, and p21, indicating that ascorbic acid relieved senescence-related G1 arrest. Analysis of AP (apurinic/apyrimidinic) sites showed that ascorbic acid treatment decreased AP site formation by 35%. We also tested the effect of hydrogen peroxide treatment, as an additional oxidative stress. Continuous treatment of $20{\mu}M$ of hydrogen peroxide from PD 40 of HEF cells resulted in premature senescence due to increased ROS level, and increased AP sites. Taken together, the results suggest that ascorbic acid extends replicative life span of HEF cells by reducing mitochondrial and DNA damages through lowering cellular ROS.

• Journal of Life Science
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• v.29 no.12
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• pp.1321-1328
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• 2019

Intermediate-wavelength solar radiation, also known as ultraviolet B (UVB: 290-320 nm) radiation, may cause premature aging and oxidative damage-dependent skin cancer in humans. UVB-induced formation of reactive oxygen species (ROS)-often a consequence of excessive exposure to these rays-could activate matrix metalloproteinases (MMPs) such as MMP-1 and MMP-3. These enzymes break down type I collagen in human fibroblasts. In this study, we assessed the antioxidant and anti-aging effects of ethyl acetate extract of blueberry (EEB). An antioxidant test in blueberries evaluated ROS production using CCD-986sk cells and DPPH assay. In order to evaluate the anti-wrinkle efficacy of blueberries, the MMP-1 production and type 1 procollagen synthesis evaluated and the expression of MMP 1, 3 were tested through Western blot and RT- PCR. EEB exhibited 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reduced the production of UVB-induced ROS. Also, EEB inhibited UVB-induced processes associated with photoaging and skin cancer, such as reduction in procollagen production and increase in MMP-1 production. More precisely, EEB (50 ㎍/ml) markedly suppressed mRNA and protein levels of MMP-1 and -3. The anti-aging effects are attributable to the antioxidant activity of EEB. These findings indicate that EEB has a protective effect against UVB-induced aging in human fibroblast cells by regulating the levels of type-1 procollagen, MMP-1, and MMP-3.

• Journal of Life Science
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• v.27 no.11
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• pp.1269-1275
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• 2017

Tenebrio molitor samples were investigated as novel biomaterials and sources of food in several recent studies. However, the insects' effects on hair growth were not sufficiently researched. To develop novel and natural materials for preventing alopecia and promoting hair growth, this study investigated the antioxidant activities and hair-growth promotion effects of TMEs. To determine the antioxidant activities, the TMEs' DPPH radical- and nitrite-scavenging activities were examined. To determine hair-growth promotion effects, proliferations of human dermal papilla cells (DPCs) and the murine fibroblast cell line NIH3T3 were evaluated by using an MTS assay. In addition, estimations were made for cell viabilities against cell death induced by dihydrotesterone (DHT) in DPCs and inhibitory effects against potassium channel blocking induced by tolbutamide (TBM) in NIH3T3 cells. The DPPH radical scavenging activity was 81.17%, and the nitrite scavenging activity was 43.69%; the activities were similar to the activities of blueberry extracts. Moreover, the TMEs promoted the proliferation of human DPCs and NIH3T3 cells, which were concentrated dependently. The TMEs prevented not only DHT-induced DPC cytotoxicity but also TBM's action as a potassium channel blocker in NIH3T3 cells. The results suggested that TME could be used as a functional therapeutic alopecia reagent, to prevent hair loss and to promote hair growth.