• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.7 no.1
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• pp.39-60
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• 2001

Fuzhengfangaitang is a prescript for inhibiting recurrence and metastasis of cancer. We had examined the anti-metatstastic effect of Fuzhengfangaitang. Furthermore, we performed the following experiments with ingredients of Fuzhengfangaitang. The purpose of this thesis is to study what ingredients of Fuzhengfangaitang have more valuable anti-cancer effects. And the results are listed below: 1. Cell Viability assay At the dose of 400$\mu$g/ml, most ingredients of Fuzhengfangaitang depressed viability of ECV-304. And especially, Scutellaria barbata D. DON$50{\mu}g/ml$ : 53.118%, $100{\mu}g/ml$: 49.092%, $200{\mu}g/ml$ : 43.765%, $400{\mu}g/ml$ : 12.747%), Polygonum bistorta L.($50{\mu}g/ml$ : 45.554%, $100{\mu}g/ml$ : 45.554%, $200{\mu}g/ml$ : 0.0%, $400{\mu}g/ml$ : 0.0%) and Psoralea corylifolia L.($50{\mu}g/ml$ : 86.591%, $100{\mu}g/ml$ : 81.307%, $200{\mu}g/ml$ : 24.801%, $400{\mu}g/ml$ : 3.111%) highly depressed cell viability more than the other ingredients. (${\alpha}$<0.05) 2. Cell Proliferation assay Proliferation assay with ingredients of Fuzhengfangaitang on ECV-304 showed that Crataegus pinnatifuda BGE ($50{\mu}g/ml$: 63.276%, $100{\mu}g/ml$ : 64.092%, $200{\mu}g/ml$ : 68.966% $400{\mu}g/ml$ : 38.517%, ED50=$296.974{\mu}g/ml$), Polygonum bistorta L.($50{\mu}g/ml$ : 83.981%, $100{\mu}g/ml$ : 86.997%, $200{\mu}g/ml$ : 58.780%, $400{\mu}g/ml$ : 26.408%), ED50=$266.725{\mu}g/ml$) and Psoralea corylifolia L.($50{\mu}g/ml$ : 103.037%, $100{\mu}g/ml$ : 82.529%, $200{\mu}g/ml$ : 2.829%, $400{\mu}g/ml$ : 0.998%), ED50=$177.369{\mu}g/ml$) depressed cell proliferation more than the other ingredients. 3. Tube Formation assay Compared with the control group, most ingredients of Fuzhengfangaitang did not remarkably inhibited the Tube Formation assay of ECV-304 at the dose of $100{\mu}g/ml$. But, Polygonum bistorta L. highly inhibited the tube formation of ECV -304 at the lower dose of $50{\mu}g/ml$. 4. Rat Aortic Ring assay In comparison with the control group, Scutellaria barbata D. DON., root of Polygonum bistorta L. and Psoralea corylifolia L. restricted the angiogenesis of the rat aortic ring at the dose of $100{\mu}g/ml$. And the other ingredients of Fuzhengfangaitang did not restricted the angiogenesis of the rat aortic ring at that dose. Especially, Polygonum bistorta L. highly inhibited the angiogenesis of the rat aortic ring at the lower dose of $50{\mu}g/ml$. From our research, the anti-angiogenic effects of the ingredients of Fuzhengfangaitang was proven. Moreover, it will be helpful for designing more effective prescription for anti angiogenesis.

• Toxicological Research
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• v.16 no.2
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• pp.173-178
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• 2000

This study was carried out to investigate cytotoxicity of several herbicides (Bentazone, Butachlor. Paraquat and Ethalfluralin) in cultured mouse NIH 3T3 fibroblasts. Tetrazolium (MTT), neutral red (NR) and sulforhodamine protein B (SRB) of the colorimetric assays were performed to evaluate the cytotoxicity on cell organelles. 2 x 10$^4$cell/$m\ell$ of NIH 3T3 fibroblast in each well of 24 multidish were cultured. After 24 hours, the cells were treated with solution (1, 25, 50 or 100 $\mu$M) of each herbicide. After the NIH 3T3 fibroblasts of all groups were cultured in the same condition for 48 hours, MTT, NR and SRB assays were performed to evaluate the cytotoxicity. The light microscopic study was carried out to examine morphological changes of cultured NIH 3T3 fibroblasts. The MTT$_{50}$ of Bentazone, Butachlor, Paraquat and Ethalfluralin were 1560.97 $\mu$M, 56.15 $\mu$M, 3138.81 $\mu$M and 1301.82 $\mu$M, respectively. The NR$_{50}$ of Bentazone, Butachlor. Paraquat and Ethalfluralin were 1763.93 $\mu$M, 45.98 $\mu$M, 1030.85 $\mu$M and 1808.29 $\mu$M, respectively. The SRB$_{50}$ of Bentazone, Butachlor. Paraquat and Ethalfluralin were 1913.38 $\mu$M, 65.30 $\mu$M, 1860.73 $\mu$M and 1086.93 $\mu$M, respectively. The morphological changes of NIH 3T3 fibroblasts showed severe degeneration in Butachlor 50 $\mu$M and 100 $\mu$M concentrations. These results indicate that Butachlor has high cytotoxicity, Bentazone, Paraquat and Ethalfluralin very weak cytotoxicity against NIH 3T3 fibroblasts.lasts.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.3
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• pp.370-374
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• 2003

Cell cracking method using a non-collision was evaluated for the possibility of new red ginseng grinding technique. Based on particle size distribution analysis by 1size shaker, the ratios of 100 mesh penetrated particles were 94.9% for hammer mill (group A) and 95.6% for cell crack (group B). The ratio of 120 mesh penetrated particle of group A was higher than that in group B. The particle size distributions for 100 mesh non-penetrated Powder between 2 groups were not significantly different, and particle size distribution analysis by laser scattering analyzer showed that the particle size ranges were 0.77~128.07 ${\mu}{\textrm}{m}$ for group A and 4.24~180.07 ${\mu}{\textrm}{m}$ for group B. The Particle size distribution in group A was more broad than that in group B. The mean particle size in group B was larger than that in group A, while the standard deviation of particle size distribution in group B was less than that in group A. Structural surface characteristics, in group A, particle size distribution was broad and the distribution curve was amorphous. The structure of individual particles was similar to unequal stone which was roughly grinded and had soft cotton-like surface. In the contrary, in group B, particle size distribution was relatively narrow and also individual size particles were ubiquitously distributed. The structure of individual particles was unequal cut stone shape.

• Preventive Nutrition and Food Science
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• v.8 no.4
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• pp.356-364
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• 2003

A methanol extract from seeds of Paeonia lactiflora (Paeoniaceae, peony) was found to possess different antiproliferative activities against four different human cancer cell lines: Hela, MCF-7, HepG2 and HT-29. Furthermore, five different methanol (20, 40, 60, 80 and 100 % MeOH) fractions obtained by fractionation of the methanol extract of the seeds on a Diaion HP-20 column exhibited differential antiproliferative effects against the above four cancer cell lines. Among five fractions, the 60 % MeOH fraction showed relatively lower antiproliferative activity on MCF-7 estrogen-sensitive breast cancer cell than the other cancer cell lines. Systematic separation of 60% the MeOH fraction by silica gel and Sephadex LH-20 columns led to the isolation of four known stilbenes, trans-resveratrol (1), trans-(+)- $\varepsilon$ -viniferin (2), gnetin H (3) and suffruticosol B (4). The four stilbenes (1∼4) exerted differential biphasic effects on cell proliferation of MCF-7 cells in a similar manner as genistein, a soybean isoflavone used as a positive reference, in the concentration range from 1.0 to 200 $\mu$M. Three stilbenes (1 ∼ 3) weakly stimulated the proliferation of MCF -7 cells at doses below 10 JIM. However, strong antiproliferative effects on MCF-7 cell were exerted by extract 1 at a dose of 200 JIM, and by 2 and 3 at doses above 25 $\mu$M. In contrast, 4 inhibited the proliferation of MCF-7 cell at a dose below 25 $\mu$M, but stimulated cell proliferation at concentrations of 50 and 100 $\mu$M. All four stilbenes (1∼4) stimulated the proliferation of ROS 17/2.8 osteoblast-like cells in the range of 10$^{-10}$ ∼10$^{-1}$ $\mu$M. Compound 1 exhibited especially potent proliferative activity, although its activity was weaker than that of genistein. Additionally, three resveratrol oligomers (2∼4) also exhibited concentration-dependently moderate proliferative activity, but less than that of 1. These results suggest that resveratrol, and its dimer and trimers from the seeds of Paeonia lactiflora may act as a phytoestrogen, but in a somewhat different manner from that of genistein.

• Nutrition Research and Practice
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• v.13 no.1
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• pp.58-63
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• 2019

BACKGROUND/OBJECTIVES: Telomeres are located at the chromosomal ends and progressively shortened during each cell cycle. Telomerase, which is regulated by hTERT and c-MYC, maintains telomeric DNA sequences. Especially, telomerase is active in cancer and stem cells to maintain telomere length for replicative immortality. Recently we reported that walnut phenolic extract (WPE) can reduce cell viability in a colon cancer stem cell (CSC) model. We, therefore, investigated the effect of WPE on telomere maintenance in the same model. MATERIALS AND METHODS: $CD133^+CD44^+$ cells from HCT116, a human colon cancer cell line, were sorted by Fluorescence-activated cell sorting (FACS) and treated with WPE at the concentrations of 0, 10, 20, and $40{\mu}g/mL$ for 6 days. Telomere lengths were assessed by quantitative real-time PCR (qRT-PCR) using telomere specific primers and DNA extracted from the cells, which was further adjusted with single-copy gene and reference DNA ($ddC_t$). Telomerase activity was also measured by qRT-PCR after incubating the PCR mixture with cell protein extracts, which was adjusted with reference DNA ($dC_t$). Transcriptions of hTERT and c-MYC were determined using conventional RT-PCR. RESULTS: Telomere length of WPE-treated cells was significantly decreased in a dose-dependent manner ($5.16{\pm}0.13$ at $0{\mu}g/mL$, $4.79{\pm}0.12$ at $10{\mu}g/mL$, $3.24{\pm}0.08$ at $20{\mu}g/mL$ and $3.99{\pm}0.09$ at $40{\mu}g/mL$; P = 0.0276). Telomerase activities concurrently decreased with telomere length ($1.47{\pm}0.04$, $1.09{\pm}0.01$, $0.76{\pm}0.08$, and $0.88{\pm}0.06$; P = 0.0067). There was a positive correlation between telomere length and telomerase activity (r = 0.9090; P < 0.0001). Transcriptions of both hTERT and c-MYC were also significantly decreased in the same manner. CONCLUSION: In the present cell culture model, WPE reduced telomere maintenance, which may provide a mechanistic link to the effect of walnuts on the viability of colon CSCs.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.1
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• pp.92-98
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• 2013

Bee Venom(below BV) has been used in alternative medicine to treat the diseases, such as pain diseases. BV contains a variety of peptides, including melittin, apamin, adolapin, MCD peptide, enzymes(i.e. PLA2), amines(i.e. histamine and epinephrine), and nonpeptide components. The two main components of BV are melittin and PLA2. The cell cytotoxic effects through the activation of PLA2 by melittin have been suggested to be the critical mechanism for the depress of cancer cell. Melittin and PLA2 have been reported to induce apoptosis and to possess anti-cancer effects and neurite outgrowth in PC12 cells. Analysis of proliferation was confirmed by MTT assay. BV decreased cell number through dose- and duration-dependent manner and these effects are apparent at a concentration of 3 ${\mu}g/ml$. To observe which signaling molecules will be activated by BV, phosphorylation of ERK, p38 MAPK, JNK and ERM were examined by Western blot analysis. To study the long term effect of BV in human gastric adenocarcinoma cell lines, the image of cells treated with BV for 4 days were obtained. BV was shown to exhibit anti-cancer activity in human gastric adenocarcinoma cell lines at a broad range of concentrations of 3 ${\mu}g/ml$. ERK, p38 MAPK and JNK were found to increase in BV treated cells. However, ERM which known to be involved in the cell death, was gradually decreased to 30minutes after addition 3 ${\mu}g/ml$ of BV. These results provide a possible BV-induced inhibitory signal for cancer proliferation that is initiated by the decrease in ERM activity. Moreover, it is likely that the activation of ERK, p38 MAPK and JNK are required for the BV-induced inhibition of cancer proliferation.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.19 no.3
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• pp.261-269
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• 2009

To evaluate the effect of antioxidant on the cytotoxicity induced by oxidative stress of reactive oxygen species (ROS) in cultured human skin melanocytes, colorimeric assay of XTT and tyrosinase activity assay were adopted after human skin melanocytes were preincubated for 2 hours in the media containing various concentrations of superoxide dismutase (SOD) before the treatment of hydrogen peroxide. Light microscopic study was carried out in same cultures. The results of this study were as follows 1. Cell viability of human skin melanocytes was significantly decreased by 30 and $40{\mu}M$ of hydrogen peroxide($H_2O_2$), respectively. 2. XTT50 was determined at $30{\mu}M$ after human skin melanocytes were treated with $10{\sim}40{\mu}M$ of hydrogen peroxide for 6 hours. 3. The cell viability of cultured human skin melanocytes pretreated with SOD was increased than that of cultured human skin melanocytes treated with $H_2O_2$ dose-dependently. 4. In tyrosinase activity of human skin melanocytes, the cell treated with SOD showed brown stain compared with $H_2O_2$ treated cells, dark stain. 5. In light microscopy, cultured human skin melanocytes exposed to $H_2O_2$ showed morphological changes such as the decreased cell number and cytoplasmic processes, compared with control. 6. In light microscopy, cultured human skin melanocytes pretreated with SOD showed the increase of cell number and cytoplasmic processes compared with $H_2O_2-treated$ group. From these results, it is suggested that oxidative stress of ROS such as $H_2O_2$ has cytotoxicity by showing the decreased cell viability, the increased tyrosinase activity and mophological changes of the decreased cell number and cytoplasmic processes. While, antioxidant like SOD was effective in the prevention of oxidative stress-mediated cytotoxicity by the increased cell viability, decreased tyrosinase activity and the protection of degenerative morphological changes in cultured human skin melanocytes.

• Korean Journal of Pharmacognosy
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• v.29 no.3
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• pp.243-247
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• 1998

A diverse panel of human tumor cell lines and a mouse melanoma cell line (B16-F1) were used for the cytotoxicity test of the nine sesquiterpene lactones with ${\beta}-methylene-{\gamma}-lactone$ group isolated from Hemisteptia lyrata, Chrysanthemum zawadskii and Chrysanthemum boreale. In the cell adhesion inhibitory activity test against B16-F1 mouse melanoma cell, hemistepcin B, cumambrin B, costunolide and tulipinolide were shown significant activities with $IC_{50}$ range of 2.2, 4.1, 0.9 and $0.3\;{\mu}g/ml$, respectively. In the cytotoxicity test against human tumor cells, the most active compound was costunolide having $IC_{50}$ values of below $0.3\;{\mu}g/ml$ against all the tested cell lines except for UACC62. Cumambrin A, hendelin and costunolide exhibited more strong activity against HCT15 and UO-31 cell lines than a positive control, adriamycin. All tested compounds showed an $IC_{50}$ values of below $5.0\;{\mu}g/ml$ against all the tested cell lines.

• Annals of Clinical Neurophysiology
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• v.8 no.1
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• pp.63-70
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• 2006

Background: Testosterone is reported to have neuroprotective effect in various neurological diseases. Recently, the mechanism involved in nitric oxide (NO)-mediated motor neuron death is under extensive investigation. The Cu/Zn-superoxide dismutase (SOD1) mutations has been implicated in selective motor neuron death of amyotrophic lateral sclerosis (ALS) and it is said to play an important role in NO-mediated motor neuron death. However, neuroprotective effect of testosterone on motor neuron exposed to NO has rarely been studied. Methods: Motor neuron-neuroblastoma hybrid cells expressing wild-type or mutant (G93A or A4V) SOD gene were treated with $200{\mu}M$ S-nitrosoglutathione. After 24 hr, cell viability was measured by MTT assay. To see the neuroprotective effect of testosterone, pretreatment with 1 nM testosterone was done 1 hr before S-nitroglutathione treatment. To study the mechanism of protective effect, $20{\mu}M$ flutamide (androgen receptor antagonist) was also pretreated with testosterone 1 hr before S-nitroglutathione treatment. Results: S-nitrosoglutathione showed significant neurotoxic effect in all three cell lines. Percentage of cell death was somewhat different in each cell line. 1 nM testosterone showed neuroprotective effect in G93A and wild-type cell line. In A4V cell line, testosterone did not showed neuroprotective effect. The neuroprotective effect of testosterone was reversed by $20{\mu}M$ flutamide. Conclusions: These results indicate that testosterone induces neuroprotection in NO-mediated motor neuron death directly through the androgen receptor. This neuroprotective effect of testosterone varies according to the types of SOD1 gene mutation. These data suggest that testosterone may be of therapeutic value against ALS.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.8
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• pp.983-988
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• 2007

Epigallocatechin gallate (EGCG), a principal antioxidant derived from green tea, is one of the most extensively investigated chemopreventive phytochemicals. However, the effect of EGCG on proliferation in MDA-MB-231 breast cancer cell is not well known. We investigated the effect of EGCG on protein and mRNA expression related to cell proliferation in MDA-MB-231 human breast cancer cell lines. We cultured MDA-MB-231 cells in the presence of 0, 5, 10 and 20 ${\mu}m$ of EGCG. EGCG significantly inhibited the cancer cell proliferation (p<0.05). In MDA-MB-231 huamn breast cancer cell, EGCG lowered $ErbB_2$ and $ErbB_3$ protein as well as mRNA expression. In addition, protein and mRNA expression of phosphorylated Akt and total Akt were significantly decreased (p<0.05). We suggest that EGCG inhibits cell proliferation through $ErbB_2$, $ErbB_3$ and Akt cell signaling.