• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.1833-1837
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• 2015

An aristolactam-type alkaloid, isolated from Orophea enterocarpa, is enterocarpam-III (10-amino-2,3,4,6-tetramethoxyphenanthrene-1-carboxylic acid lactam). It is cytotoxic to various human and murine cancer cell lines; however, the molecular mechanisms remain unclear. The aims of this study were to investigate cytotoxic effects on and mechanism (s) of human cancer cell death in human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells compared to normal murine fibroblast NIH3T3 cells. Cell viability was determined by MTT assay to determine $IC_{10}$, $IC_{20}$ and $IC_{50}$ levels, reactive oxygen species (ROS) production with 2',7'-dichlorohydrofluorescein diacetate and the caspase-3, -8 and -9 activities using specific chromogenic (p-nitroaniline) tetrapeptide substrates, viz., DEVD-NA, IETD-NA and LEHD-NA and employing a microplate reader. Mitochondrial transmembrane potential (MTP) was measured by staining with 3, 3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and using flow cytometry. The compound was cytotoxic to HepG2 and MDA-MB-231 cells with the $IC_{50}$ levels of $26.0{\pm}4.45$ and $51.3{\pm}2.05{\mu}M$, respectively. For murine normal fibroblast NIH3T3 cells, the $IC_{50}$ concentration was $81.3{\pm}10.1{\mu}M$. ROS production was reduced in a dose-response manner in HepG2 cells. The caspase-9 and -3 activities increased in a concentration-dependent manner, whereas caspase-8 activity did not alter, indicating the intrinsic pathway activation. Enterocarpam-III decreased the mitochondrial transmembrane potential (MTP) dose-dependently in HepG2 cells, suggesting that the compound induced HepG2 cell apoptosis via the mitochondrial pathway. In conclusion, enterocarpam-III inhibited HepG2 and MDA-MB-231 cell proliferation and induced human HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway and induction of caspase-9 activity.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.5
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• pp.2661-2665
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• 2016

Silibinin is a natural polyphenol with high antioxidant and anticancer properties. In this study, its influence on two of the most commonly employed human breast cancer cell lines, MCF-7 and T47D, and one non-malignant MCF-10A cell line, were investigated and compared. Cell viability, the cell cycle distribution and apoptosis induction were analyzed by MTT and flow cytometry, respectively. The effect of silibinin on PTEN, Bcl-2, P21, and P27 mRNAs expression was also investigated by real-time RT-PCR. It was found that silibinin caused G1 cell cycle arrest in MCF-7 and MCF-10A cells but had no effect on the T47D cell cycle. Silibinin induced cytotoxic and apoptotic effects in T47D cells more than the MCF-7 cells and had no cytotoxic effect in MCF-10A cells under the same conditions. Silibinin upregulated PTEN in MCF-7 and caused slightly increased P21 mRNA expression in T47D cells and slightly increased PTEN and P21 expression in MCF-10A cells. Bcl-2 expression decreased in all of the examined cells under silibinin treatment. P27 mRNA expression upregulated in T47D and MCF-10A cells under silibinin treatment. PTEN mRNA in T47D and P21 and P27 mRNAsin MCF-7 were not affected by silibinin. These results suggest that silibinin has mostly different inhibitory effects in breast cancer cells and might be an effective anticancer agent for some cells linked to influence on cell cycle progression.

• International Journal of Oral Biology
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• v.36 no.1
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• pp.13-21
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• 2011

Chios gum mastic (CGM) is produced from Pistiacia lentiscus L var chia, which grows only on Chios Island in Greece. CGM is a kind of resin extracted from the stem and leaves, has been used for many centuries in many Mediterranean countries as a dietary supplement and folk medicine for stomach and duodenal ulcers. CGM is known to induce cell cycle arrest and apoptosis in some cancer cells. This study was undertaken to investigate the alteration of the cell cycle and induction of apoptosis following CGM treatment of HL-60 cells. The viability of the HL-60 cells was assessed using the MTT assay. Hoechst staining and DNA electrophoresis were employed to detect HL-60 cells undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity analyses were also employed. CGM treatment of HL-60 cells was found to result in a dose- and time-dependent decrease in cell viability and apoptotic cell death. Tested HL-60 cells showed a variety of apoptotic manifestations and induced the downregulation of G1 cell cycle-related proteins. Taken collectively, our present findings demonstrate that CGM strongly induces G1 cell cycle arrest via the modulation of cell cycle-related proteins, and also apoptosis via proteasome, mitochondrial and caspase cascades in HL-60 cells. Hence, we provide evidence that a natural product, CGM could be considered as a novel therapeutic for human leukemia.

• Korean Journal of Acupuncture
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• v.29 no.2
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• pp.271-289
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• 2012

Objectives : Ganoderma lucidum(Ganoderma or lingzhi, 靈芝) is a well-known oriental medical mushroom containing many bioactive compounds. The possible mechanisms involved in its effects on cancer cells remain to be elucidated. In the present study, the anti-proliferative and apoptotic activities of the G. lucidum ethanol extract(GEE), in AGS human gastric cancer cells were investigated. Methods : It was found that exposure of AGS cells to GEE resulted in the growth inhibition in a dose and time dependent manner as measured by trypan blue count and MTT assay. The anti-proliferative effect of GEE treatment in AGS cells was associated with morphological changes and formation of apoptotic bodies, and the flow cytometry analysis confirmed that GEE treatment increased the populations of apoptotic-sub G1 phase. Growth inhibition and apoptosis of AGS cells by GEE were connected with a concentration and time-dependent up-regulation of tumour necrosis factor-related apoptosis-inducing ligand(TRAIL) expression. Results : The levels of XIAP and survivin expression, members of IAP family proteins, were gradually down-regulated by GEE treatment. However other members of IAP family proteins such as cIAP-1 and cIAP-2 remained unchanged in GEE-treated AGS cells. GEE treatment also induced the proteolytic activation of caspase-3, caspase-8 and caspase-9 and a concomitant degradation of poly(ADP-ribose) polymerase(PARP) protein, a caspase-3 substrate protein. Additionally, GEE-induced apoptosis was associated with the inhibition of Akt activation in a concentration and time-dependent manner, and pre-treatment with LY294002, a phosphoinositide 3-kinase(PI3K)/Akt inhibitor, significantly increased GEE-induced growth inhibition and apoptosis. Conclusions : Therefore, G. lucidum has a strong potential as a therapeutic agent for preventing cancers such as gastric cancer cells.

• Analytical Science and Technology
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• v.9 no.1
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• pp.26-34
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• 1996

A method to determine the trace amount of Se and Bi in the scalp hair using the hydride generation inductively coupled plasma atomic emmission spectrometry was studied. The optimum operation conditions of ICP for hydride generation are 0.6~0.8L/min for the carrier gas flow rate, and 6mm above the induction coil for the observation height. Hydrochloric acid concentrations for the optimum hydride generation conditions were greater than 1.5M when 2.5% $NaBH_4$ and NaOH were used, and greater than 0.5M when 2.5% $NaBH_4$ and 0.1% NaOH were used. Severe interference effects are observed from transition metals such as Cu and Ni, and they could be circumvented by the coprecipitation with lanthanum hydroxide.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.6
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• pp.1427-1432
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• 2003

This study was carried out to investigate the antimicrobial and anticancer effects of Galla Rhois (GR) on pathogens isolated from oral and KB human oral epidermoid carcinoma cells. Their antimicrobial activities were tested against Streptococcus mutans (S. mutans), Staphylococcus aureus (S. aureus), Enterobacter aerogenes (E. aerogenes), Escherichia coli (E. coil) and Candida albicans (C. albicans). GR powder has the antimicrobial activity against C. albicans, S. mutans and S. aureus. The extracts of water and ethanol have the antimicrobial activity against S. sureus and C. albicans. The water extract showed inhibitory activity against the growth and pH of above mentioned reference microorganisms. The water extract of GR declined cell viability in a dose dependent manner. DNA flow cytometric analysis showed that population of sub-G/sub 0//G₁, phase of cell cycle was increased by GR extract treatment in a dose dependent manner. Western blot analysis revealed that Caspase-3 was reduced by GR extract treatment. These result suggested that GR has the effect of antimicrobial on pathogens isolated from oral, and also, has anticancer effect that associated with the induction of apoptosis in a dose dependent manner in KB human oral epidermoid carcinoma cells. It may be GR-induced apoptosis was mediated via activation of Caspase-3.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2005.06a
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• pp.942-945
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• 2005

Micron-sized metal wires are widely used in industries such as filtration, catalyst and composite materials, etc. In the wire drawing process, the die that is used conventionally is an effective and, at the same time, sensitive component. However, a typical array of the dies has caused many problems in the wire drawing process, e.g., large frictional force on the interface between wire and the resulting high heat generation, precise adjustment of the dies, extended cooling system, die abrasion, etc.. Because of these problems, there have been many works that are aiming at improving the efficiency of wire drawing process by analyzing the die geometry and by applying advanced die material to prolong the die life or even at developing a dieless wire drawing system. This paper is dealing with developing a new wire drawing system that is applicable to reduce the wire drawing steps with high draw ratio. The new wire drawing system does not use the dies, but use the self-induced heater that works on the basis of the resonant phenomenon of wire material. The electromagnetic wave is the heating source. The results of the study on the diameter reduction and microwave flow analysis show that the heating effectiveness of the wire is influenced by the energy distribution in the microwave propagation chamber. We can obtain diameter-reduced wires by using microwave in the dieless drawing process. Microwave as a heating source is capable of producing wires without applying dies in wire drawing process.

• Journal of Photoscience
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• v.5 no.4
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• pp.169-174
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• 1998

Photosynthetic responses to dehydration were examined by the simulataneous measurement of O2 evolution and chlorophyll (Chl) fluorescence in green pepper leaves. Dehydration was induced by immersing the plant roots directly in the Hoagland solution containing varying concentration (2-30%) of polyethylene glycol(PEG-6000) . Water potential of the leaf was decreased time-and concentation -dependently by PEG-treatment. The decrease in water potential of leaf was correlated with the decrease in both the maximal photosynthesis (Pmax) and quantum yield of O2 evolution, but Pmax dropped more rapidly than quantum yield at all water deficit conditions tested. However, Chl fluorescence parameters were not affected much. Dehydration did not change the initial fluorescence (Fo) and maximum photochemical efficiency(Fv/Fm) of photosystem(PS) II. Both the photochemical quenching (qP) and non-photochemical quenching(NPQ) were not changed by dehydration under low PFR(50 $\mu$mols m-2s-1 ). In contrast, under high PFR(270$\mu$mols m-2s-1)qP was slightly decreased while NPQ was greatly increased. The fast induction kinetics of Chl fluroecence showed no change in Chl fluorescence pattern by dehydration at high PFR (640 $\mu$mols m-2s-1 ), but exhibited a significant drop in peak level(Fp)at low PRFR (70$\mu$mols m-2s-1 ). PS I oxidation and reduction kinetics revealed normal reduction but delayed oxidation to P-700+, suggesting no lesionin electron flow from PSII to PSI , but impaired electron transport to NADP+,These results suggest that water stress caused by PEG-treatment results in the reduction of photosynthesis, promarily due to the reducted electron trasport from PSI to NADP+ or hampered subsequent steps involving Calvin Cycle.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.4
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• pp.745-755
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• 2002

To examine the effects of Yunpyesan on the cell proliferation of A549 human lung carcinoma cell line, we performed various experiments such as dose-dependent effect of Yunpyesan on cell proliferation and viability, morphological changes, quantification of apoptotic cell death and alterations of apoptosis/cell cycle-regulatory gene products. Yunpyesan declined cell viability and proliferation in both a dose- and a time-dependent manner. The anti-proliferative effect by Yunpyesan treatment in A459 cells was associated with morphological changes such as membrane shrinking and cell rounding up. Yunpyesan Induced apoptotic cell death in a time-dependent manner, which was associated with degradation of poly-(ADP-ribose) polymerase (PARP), an apoptotic target protein, without alterations of the balance between Bcl-2 and Bax expressions. DNA flow cytometric histograms showed that population of G1 phase of the cell cycle was increased by Yunpyesan treatment in a dose-dependent manner. Western blot analysis revealed that cyclin D1 and A were reduced by Yunpyesan treatment, whereas cyclin dependent kinase (Cdk) inhibitor p27 was markedly increased in a time-dependent fashion. The level of tumor suppressor p53 proteins was also increased by Yunpyesan treatment and its increase might be linked to increase of Cdk inhibitor p27. In addition, Mdm2, negative regulator of p53, was down-regulated by Yunpyesan treatment. Since the expression of retinoblastome protein (pRB), a key regulator of G1/S progression, was reduced by Yunpyesan treatment, we supposed that phosphorylation of pRB might be also blocked. The present results indicated that Yunpyesan-induced inhibition of lung cancer cell proliferation is associated with the induction of apoptosis and the blockage of G1/S progression.

• The Journal of Korean Medicine
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• v.29 no.2
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• pp.7-20
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• 2008

Objectives: The purpose of this study wasto examine the effects of Kamijihwang-tang (KJHT) extracts on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods: C57BL/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (four times a week) for asthma induction. Two experimental groups were treated with different concentrations of KJHT (400 mg/kg and 200 mg/kg) extracts and cyclosporine A (10 mg/kg) for the later 8 weeks. At the end of the experiment, the mice lung, PLN and spleen were removed and immune cells were analyzed by flow cytometer, IL-5, IL-13, eotaxin-2, $TNF-{\alpha}$ were analyzed by real-time PCR, serum histamine was analyzed by ELISA kit. Results: $CD3^+$, $CD3e^-/CCR3^+$, $CD3e^+/CD69^+$, $CD4^+/CD25^+$, $B220^+/IgE^+$, and $CD3e^+/DX5^+$ cells in lung, PLN, and spleen of the KJHT group (400 mg/kg) decreased compared with that of the control group. $CD3e^+/CD69^+$, $CD4^+/CD25^+$, and $CD3e^+/DX5^+$ cells in lung, PLN, and spleen of the KJHT group (200 mg/kg), CD3+, $CD3e^-/CCR3^+$ cells in lung and PLN of the KJHT group (200 mg/kg) and $B220^+/IgE^+$ cells in lung and spleen of the KJHT group (200 mg/kg) decreased compared with that of the control group. mRNA expression of IL-5, IL-13, eotaxin-2, and $TNF-{\alpha}$ in lung tissue of the KJHT groups (400 mg/kg and 200 mg/kg) decreased compared with that of the control group. Histamine in serum of the KJHT group (400 mg/kg) decreased compared with that of the control group. Conclusions: These results suggest that KJHT can be utilized effectively in treating asthma because it significantly reduces inflammatory cells and cytokines.