• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.7
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• pp.819-824
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• 2007

The present study describes the cytotoxic effect of Artemisia fukudo extracts. The extract from A. fukudo by 80% ethanol was fractionated with n-hexane, dichloromethane, ethylacetate, and butanol in serial. The cytotoxicity of A. fukudo extracts was examined for its effect on the growth of HL-60 cells by the colorimetric 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. In addition, we used the HL-60 cells to see what effects the A. fukudo extracts will have on apoptosis of cancer cells. We checked the cell activity, cell morphologic changes, DNA fragmentation, and DNA content after 24 hr incubation with administering 25 ${\mu}g/mL$ of the A. fukudo extracts. In the treatment of the low concentration of n-hexane and dichloromethane fractions, the survival rate of HL-60 cells is lower than that of the control. The laddering-pattern DNA fragmentation was observed in the treatment with n-hexane and dichloromethane fractions. The DNA content of the cells apoptosis measured as the density of sub-$G_{1}$ hypodiploid cells by flow cytometric analysis. The number of sub-$G_{1}$ hypodiploid cells increased in the treatment with n-hexane and dichloromethane fractions. These fractions obstructed the cell cohesion and caused the blebbing of the cell membrane and fragmentation of the nucleus, both of which are symptoms of apoptosis. These results suggest that A. fukudo has a great potential value as food additives, medicinal supplements for patients with chronic diseases, and preventive measures against cancer.

• Journal of the East Asian Society of Dietary Life
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• v.24 no.3
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• pp.351-358
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• 2014

Pumpkin seed oil (PSO) was investigated for its parasite elimination activity and efficacy in treating disorders of the prostate gland and urinary bladder. We confirmed the composition of PSO and identified its ability to improve vessels. Gas chromatography coupled with mass spectrometric (GC-MS) system was used for PSO composition analysis. Cytotoxicity and cell proliferation were confirmed by Cell Counting Kit-8 (CCK-8) assay. Nitric oxide(NO) production was measured with Griess reagent, and intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 mRNA expression levels were measured by reverse transcription polymerase chain reaction (RT-PCR). As a result, PSO revealed the presence of several components such as linoleic acid, oleic acid, palmitic acid and stearic acid. Cytotoxic effects of PSO were not observed, and PSO increased nitric oxide production in human umbilical vein endothelial cells (HUVEC). Additionally, TNF-${\alpha}$-induced cell proliferation and ICAM-1 expression in HUVEC were inhibited by PSO treatment, whereas VCAM-1 expression was not significantly reduced. Taken together, these results show that PSO is worthy of study as a candidate food material for improvement of vascular disease.

• Korean Journal of Food Science and Technology
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• v.37 no.2
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• pp.221-227
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• 2005

Apoptosis of Prunus salicina Lindl. cv. Soldam, which possesses hematopoiesis, osteoporosis prevention, and antimutagenic effects, at different growth stages was evaluated. Cytotoxic effect of acetone extracts of immature fruits against various tumor cell lines was higher than that of mature fruits, particularly in hormone-independent human breast cancer, MDA-MB-231 cell line. Immature fruit extract increased expression level of pro-apoptotic protein Bax and reduced that of anti-apoptotic protein Bcl-2, and stimulated caspase-3 activity in MDA-MB-231 cells. Results suggest immature fruit of P. salicina Lindl. cv. soldam to be natural source for development of functional food and medical agents to prevent human breast cancer.

• Journal of Life Science
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• v.16 no.4
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• pp.659-663
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• 2006

The extracts of Plocamium telfairiae using several solvents with different polarities were prepared and their growth inhibitory effects were examined on the human cancer cells. We investigated the cytotoxic effects of P. telfairiae extracts on HT-29 cells by the MTT reduction assay and examining the morphological change under the inverted microscope. Among three extracts, the methanol extract showed the strongest inhibitory effect on the growth of HT-29 cells. The methanol extract was further fractionated sequentially with n-hexane, diethyl ether, ethyl acetate, and aqueous for purifying crude methanol extract. The n-hexane layer among the fractioned layers showed remarkable inhibitory activity on the growth of HT-29 cells. Moreover n-hexane layer showed the notable growth inhibition effects with a dose-dependent manner against SW620, HeLa, and MCF-7 cells as well as HT-29 cells. These results indicated that P. telfairiae extracts may be contained bioactive materials with inhibitory effect on the growth of human cancer cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1341-1348
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• 2009

Hoechunyanggyeok-san (HYS) is a traditional oriental herbal medicine widely used for treating inflammatory disorders. Although there are numerous clinical results of HYS reported in the literature of oriental hebal medicine, it has been rarely conducted to evaluate the immuno-biological activity. The present study was conducted to examine the anti-inflammatory effects of HYS extract (HYSE) in vivo and in vitro. To determine the cytotoxic concentration of HYSE, cell viability was tested by MTT assay. All four doses of HYSE (0.01, 0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity during the entire experimental period. In order to measure NO levels in culture medium, the cells were treated with $1\;{\mu}g/ml$ of LPS 1h before adding HYSE for 24 h and then culture medium were reacted with Griess reagent. Increased NO production and iNOS expression were detected in LPS-activated cells compared to control. However, these increases were dose-dependently attenuated by treatment with HYSE. LPS plays a key role in leading to the massive production of pro-inflammatory cytokines such as TNF-$\alpha$, IL-$1{\beta}$ and IL-6 in macrophages. Thus, we next determined the levels of these cytokines. HYSE reduced the elevated production of TNF-$\alpha$, IL-$1{\beta}$ and IL-6 by LPS. Moreover, the effects of HYSE were in a dose-dependent manner. In vivo, histopathological study, HYSE effectively inheefed the increases of hind paw skin thicknesses and inflammatory cell infiltrations induced by carrageenan treatment. It, therefore, considered that HYSE will be favorably inheefed the acute edematous inanner. In s. These findings showed that HYSE could have anti-inflammatory effects through the reduction of NO and inflammatory cytokines in macrophage. Furthermore, the reduction of carrageenan-induced paw oedema by HYSE helps to understand its actions on inflammatory conditions.

• The Korean Journal of Physiology and Pharmacology
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• v.10 no.5
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• pp.289-295
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• 2006

Sodium fluoride (NaF) has been shown to be cytotoxic and elicit inflammatory response in human. However, the cellular mechanisms underlying NaF-induced cytotoxicity in periodontal tissues have not yet been elucidated. This study is aimed to investigate the mechanisms of NaF-induced apoptosis in human gingival fibroblast (HGF). NaF decreased the cell viability of HGF in a dose- and time-dependent manner. NaF gave rise to apoptotic morphological changes including cell shrinkage, chromatin condensation, and DNA fragmentation. However, NaF did not affect the production of ROS. In addition, NaF augumented cytochrome c release from mitochondria into the cytosol, and enhanced caspase -9 and -3 activities., cleavage (85 kDa fragments) of poly (ADP-ribose) polymerase (PARP) and upregulation of voltage-dependent anion channel (VDAC) 1. These results demonstrated that NaF-induced apoptosis in HGF may be mediated with mitochondria. Furthermore, NaF elevated caspase-8 activity and upregulated Fas-ligand (Fas-L), suggesting involvement of death receptor mediated pathway in NaF-induced apoptosis. Expression of Bcl-2, an anti-apoptotic Bcl-2 family, was downregulated, whereas expression of Bax, a pro-apoptotic Bcl-2 family, was not affected in NaF-treated HGF. These results suggest that NaF induces apoptosis in HGF through both mitochondria- and death receptor-mediated pathway mediated by Bcl-2 family.

• Journal of Life Science
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• v.26 no.11
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• pp.1320-1329
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• 2016

The present study examined the cytotoxic effects of 1, 2, 3, 4, 6-penta-O-galloyl-${\beta}$-D-glucose (PGG), known as the pentahydroxy gallic acid ester of glucose, in the various human cancer cell lines (A-549, MDA-MB-231, U87-MG, MCF-7 and PANC-1), normal MRC-5 fetal fibroblasts, and dental papilla tissue- derived mesenchymal stem cells (DPSCs). Significantly (p<0.05) lower half maximal inhibitory concentration ($IC_{50}$) values were observed in the A-549 and MDA-MB-231 cells showing a high proliferation capacity, compared with other cancer and normal cell lines with a relatively low proliferation capacity. The population doubling time (PDT) was significantly (p<0.05) higher in the $10{\mu}M$ PGG-treated cell lines than those of untreated control cell lines. The present study demonstrated that the $IC_{50}$ value increases proportionally to the extending PDT. A high cell number with senescence-associated ${\beta}-galactosidase$ activity was also observed in the $10{\mu}M$ PGG-treated cells compared with those of untreated control cells. Moreover, the level of telomerase activity was significantly (p<0.05) decreased with $10{\mu}M$ PGG treatment, especially in A-549 and MDA-MB-231 cells showing a high proliferation capacity. Based on these observations, PGG could serve as a potent agent for cancer chemotherapy, as its treatment was more effective in cells with a high proliferation capacity.

• 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 Microbiology and Biotechnology
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• v.27 no.12
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• pp.2156-2164
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• 2017

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

• Journal of Biomedical Engineering Research
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• v.16 no.3
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• pp.331-336
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• 1995

A composite material was produced as an artificial bone substitute which is gradually degrAded and replaced by the regenerated natural bones after implantation. To detect the effect of the material on the cell's activity, the composite specimens were placed in MEMs and incubated at $37^{\circ}C$ for one week. Human uterus cervical cancer origin HeLa 3 cells and mouse subcutaneous origin L929 cells were cul- tured in the specimen dissolved MEMs for 5 days to investigate cytotoxicity via cell growth rates. ${Na_2}^{51}CrO_4$ solution was added to the media, to label the HeLa 53 cells, and the released amount of $^{51}Cr$ was measured by a $\gamma$-counter. On the cell growth investigation, no significant cytotoxic phenomena were revealed in both HeLa S3 and L929 cell cultures. On the released 51CR from the incubated HeLa 53 cells, no significant cell degeneration was observed from the composite embedded MEMs.