• The korean journal of orthodontics
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• v.23 no.2 s.41
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• pp.229-248
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• 1993

[ $Interferon-\gamma$ ] has been suggested as a cytokine of connective tissue stabilizer. In addition, it has also been demonstrated that this cytokine inhibited bone remodeling activities of the bone derived cells. In order to illuminate the effects of this cytokine in orthodontic force induced bone remodeling, it was administered to primary cultured periodontal ligament cells which have been known to have some osteoblast like characteristics. $Interferon-\gamma$ slightly decreased $[^3H]thymidine$ incorporation rate without a significant change in the total cellular DNA content up to 1000 U/ml, which meant these doses were not cytotoxic to the cell. Total protein synthesis was not influenced by various concentration of interferon-y whether it was determined by the $[^3H]proline$ incorporation rate or by the Lowry smethod. The effect of $interferon-\gamma$ on the individual protein was, however, differential, ie, it increased $[^3H]proline$ incorporation into the noncollagenous protein marginally, while it decreased $[^3H]proline$ incorporation into the collagen, so that it caused dose-dependent suppression of the relative collagen synthesis. On the contrary, the fibronectin synthesis determined by the ELISA was increased by 1000 U/ml of $interferon-\gamma$. The differential effects of the interferon-y on the collagen and fibronectin synthesis exhibited not only their protein level but also the steady state mRNA level. $Interferon-\gamma$ decreased steady state level of ${\alpha}1(I)$ procollagen mRNA significantly, while showing no significant changes in the fibronectin mRNA level. In addition to this, it was also found that indomethacin did not affect on the $interferon-\gamma$ induced collagen decrease in this cell, which meant prostaglandins were not involed in the process of $interferon-\gamma$ induced collagen decrease. So it can be concluded that the incubation of periodontal ligament cells with 1000 U/ml of $interferon-\gamma$ for 24 hr showed differential effects on the type I collagen and fibronectin gene expression. The decrease in relative collagen synthesis in the protein level was related with decrease in the steady state level of mRNA, while the increase in the fibronectin synthesis in the protein level was not correlated with the mRNA level.

• Journal of Chest Surgery
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• v.34 no.6
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• pp.447-453
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• 2001

Background: Although pulmonary resection is the standard approach for the management of pulmonary metastases from soft tissue sarcoma, most of them are unresectable and chemotherapy remains the only option. The effectiveness of the cytotoxic drugs may be limited by the toxicities that occur before the therapeutic dose is reached. The regional administration of doxorubicin using pulmonary arterial perfusion in a rodent model can produce 10 to 25 times higher concentrations in the lung than systemic administration with minimal systemic toxicities. However, it is unclear whether a high concentration of doxorubicin has beneficial effects for killing cancer cells. Material and Method: We studied this to evaluate the dose-dependent cytotoxic and apoptotic effects of doxorubicin on methylcholanthrene-induced rat fibrosarcoma(MCA) cells. This study examined the cytotoxicity and apoptosis-related gene expressions(Fas, FasL, Bax, caspase 1, caspase 2, caspase 8, Bcl-2, Bcl-xL, Bcl-xS) in MCA cells after 24 hours exposure to various concentrations of doxorubicin such as 1, 5, 10, 50, and 100 $\mu$M. Result: Dose-dependent cytotoxicity was observed after 24 hours exposure to doxorubicin. However, peak apoptosis after 24 hours exposure was observed at 5 $\mu$M of doxorubicin. Above 5 $\mu$M, apoptotic activity was decreased with dose-increment. All mRNA levels of apoptosis-related genes after 24 hours exposure were up-regulated above the control level at 1 $\mu$M of doxorubicin and then decreased by doxorubicin dose-increment except caspase 8, which showed higher levels than the control level at 5 $\mu$M. Apoptosis-related protein levels were highest at 1 $\mu$M of doxorubicin and then decreased by doxorubicin dose-increment. However, Bax and Bcl-xL proteins steadily showed higher levels than the control throughout the different concentrations of doxorubicin. Conclusion: These results suggest that apoptosis is the main cytotoxic mechanism in low concentrations of doxorubicin in MCA cells and apoptosis-related genes, such as Bax, caspase 8, and Bcl-xL, are involved. At high concentrations, doxorubicin still can kill MCA cells, even when apoptosis is inhibited, and have its propriety for achieving much cytotoxicity against MCA cells.

• Archives of Pharmacal Research
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• v.27 no.4
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• pp.402-406
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• 2004

This research team found in previous studies, that the ginseng saponin metabolite IH901 induces apoptosis in HepG2 cells via a mitochondrial-mediated pathway, which resulted in the activation of caspase-9 and subsequently of caspase-3 and -8. Based on these results, the involvement of the Fas/Fas ligand (FasL) death-receptor pathway, in IH901-induced apoptosis in HepG2 cells, was investigated. Levels of Fas and the Fas ligand (FasL) mRNA or protein were not increased by IH901, rather they were decreased significantly at 18 h post treatment. Soluble FasL (sFasL) was detectable by immunoprecipitation analysis En the medium of HepG2 cells treated with IH901. Increased levels of sFasL were inversely correlated with the levels of FasL. Preincubation of HepG2 cells with antagonistic anti-Fas antibody showed little protective effect, if any, on IH901-induced cell death. At a $30{\mu}M$ (24 and 48 h) and $40{\mu}M$ (24 h) concentration of IH901, the cytotoxic effect of IH901 was less then $50\%$, anti-Fas antibody prevented IH901-induced cell death. However, at a $60{\mu}M$ (24 and 48 h) and $40{\mu}M$ (48 h) concentration of IH901, cell death rates were about $80\%$ or more and most of the chemopreventive and chemotherapeutic effects of IH901 were manifested. Blocking the Fas receptor did not influence IH901-induced cell death. These results indicate that the Fas/FasL system is engaged, but not required for IH901-induced cell death, at pharmacologically significant concentrations.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.1
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• pp.57-62
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• 2009

Smilacis Chinae Radix has been used as an anti-inflammatory agent. This study was performed to anti-inflammatory and MAP kinase signaling pathway in vitro. Experimental studies were obtained by measuring the Cytotoxicity, production of NO, PGE2, TNF-$\alpha$ and protein level of catalase, SOD, MAP kinase, The results were summarized as follows: Smilacis Chinae Radix was not cytotoxic effects against Raw264.7 and HEK293 cells. Concentration of $100{\mu}g/m{\ell}$ Smilacis Chinae Radix inhibited the production of NO in the Raw264.7 cell stimulated with LPS. All concentrations of Smilacis Chinae Radix not significantly inhibited the production of PGE2 in the Raw264.7 cell stimulated with LPS. All concentrations of Smilacis Chinae Radix did not inhibit the production of TNF-$\alpha$ in the Raw264.7 cell stimulated with LPS. Smilacis Chinae Radix has not effect of blocking NF-${\kappa}B$ into nucleus in LPS-induced macrophage Raw264.7 cell. Smilacis Chinae Radix has the effect of Cytoprotection through activation of ERK and inhibition of p38 and JNK. Accordingly the results show Smilacis Chinae Radix could induce anti-inflammation and Cytoprotection effects against In vitro, but it needs more research on the precise mechanism of such effects.

• Journal of Acupuncture Research
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• v.18 no.4
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• pp.101-115
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• 2001

Objective : To characterize the antitumorigenic potential of Apamin, one of the major components of bee venom, its effects on cell proliferation and the mitogen-activated protein kinase (MAPK) signal transduction pathway were characterized using the human melanoma cell line SK-MEL-2. Methods & Results : Cell counting analysis for cell death demonstrated that consistent with a previous results, SK-MEL-2 cells treated with $0.5-2.0{\mu}g/ml$ of Apamin showed no recognizable cytotoxic effect whereas detectable induction of cell death was identified at concentrations over $5.0{\mu}g/ml$. [3H]thymidine incorporation assay for cell proliferation demonstrated that DNA replication of SK-MEL-2 cells is inhibited by Apamin in a dose- and time-dependent manner. To explore whether Apamin-induced growth suppression is associated with the MAPK signaling pathway, phosphorylation of Erk, a function mediator of MAPK growth-stimulating signal, was examined Western blot assay using a phospho-specific Erkl/2 antibody. A significant increase of Erkl/2 phosphorylation level was observed in Apamin-treated cells compared with untreated control cells. Qantitative RT-PCR analysis revealed that Apamin inhibit expression of MAPK downstream genes such as c-Jun, c-Fos, and cyclin D1 but not expression of MAPK pathway component genes including Ha-Ras, c-Raf-1, MEK1, and Erk. Conclusion : It is strongly suggested that the antitumorigenic activity of Apamin might result in part from its inhibitory effect on the MAPK signaling pathway in human melanoma cells SK-MEL-2.

• The Korea Journal of Herbology
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• v.23 no.2
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• pp.169-178
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• 2008

Objectives : Inflammation is important event in the development of vascular diseases including hypertension, atherosclerosis, and restenosis. Injinho-tang(IJHT) has been used as a traditional Korean herbal medicine since ancient times, and today it is widely used as a medication for jaundice associated with inflammation of the liver. The aim of this study was to determine whether IJHT and its components inhibit production of nitrite, an index of NO, and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Methods : Cytotoxic activity of IJHT and its components on RAW 264.7 cells was using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : IJHT and its components significantly inhibited the LPS-induced NO production and iNOS expression accompanied by an attenuation of tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-6 (1L-6), IL-$1{\beta}$ and monocyte chemoattractant protein-1 (MCP-1) formation in macrophages. Conclusions : IJHT and its components inhibit LPS-induced inflammation via decreasing cytokines production. These results indicate that IJHT and its components have potential as an anti-inflammation and anti-artherosclerosis agent.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.3
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• pp.808-818
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• 2004

This study was designed to elucidate the synergistic cytotoxic mechanisms of the cotreatment of Paljintanghabhwajeoghwan (Paljin) and As₂O₃ in human lung cancer cell line, H-460. The combination of Paljin and As₂O₃ synergistically augmented the cytotoxicity of Paljin and As₂O₃ in H-460 cells. The nature of cytotoxicity was revealed as apoptosis which characterized by chromatin condensation and fragmentation in DAPI staining. Mitochodrial membrane potential transition was observed in H-460 cells treated with Paljin and As₂O₃. The apoptotic cytotoxicity of Paljin and As₂O₃ was accompanied by the cleavage of PARP and ICAD. Of note, pro-apoptotic Bak protein was obviously increased. However, the expression of p53 was not affected by the cotreatment of Paljin and As₂O₃. In addition, the expression of DR5 was increased by the cotreatment of Paljin and AS203. This results suggest that the synergistic cytotoxicity of the cotreatment of Paljin and As₂O₃ might be caused by the changes of the expression levels of a lots proteins, such as PARP, ICAD, Bak, DR5, which play pivotal roles in survival or death of cells.

• International Journal of Industrial Entomology and Biomaterials
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• v.26 no.2
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• pp.89-94
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• 2013

Entomopathogenic fungal species have been investigated for their potential use as biological control agents owing to their natural role as insect pathogens. These fungi produce a wide range of secondary metabolites with high therapeutic values, such as antibiotics and cytotoxic substances. To evaluate the antibacterial activity of entomopathogenic fungi, 10 isolates from Korean soil were selected and tested for their activity against Escherichia coli by using fungal culture filtrates. Antibacterial activity was assessed using a two-step process: (1) a screening assay for the selection of fungal isolates and (2) a quantitative assay to evaluate the activity of select fungi. Although 4 fungal isolates were selected through the screening assay, only 3 fungal isolates, from Beauveria bassiana and Metarhizium anisopliae, showed high antibacterial activity according to the quantitative assay. The antibacterial activity of selected fungal culture filtrates was stable when exposed to heat and proteolytic enzyme treatments, which indicated that the antibacterial compound is not a protein. These entomopathogenic fungal metabolites might be useful as a source for bacterial control and in the pharmaceutical industry.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.9
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• pp.3981-3986
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• 2014

Ganoderma lucidum polysaccharides (GLP) extracted from Ganoderma lucidum have been shown to induce cell death in some kinds of cancer cells. This study investigated the cytotoxic and apoptotic effect of GLP on HCT-116 human colon cancer cells and the molecular mechanisms involved. Cell proliferation, cell migration, lactate dehydrogenase (LDH) levels and intracellular free calcium levels ($[Ca^{2+}]i$) were determined by MTT, wound-healing, LDH release and fluorescence assays, respectively. Cell apoptosis was observed by scanning and transmission electron microscopy. For the mechanism studies, caspase-8 activation, and Fas and caspase-3 expression were evaluated. Treatment of HCT-116 cells with various concentrations of GLP (0.625-5 mg/mL) resulted in a significant decrease in cell viability (P< 0.01). This study showed that the antitumor activity of GLP was related to cell migration inhibition, cell morphology changes, intracellular $Ca^{2+}$ elevation and LDH release. Also, increase in the levels of caspase-8 activity was involved in GLP-induced apoptosis. Western blotting indicated that Fas and caspase-3 protein expression was up-regulated after exposure to GLP. This investigation demonstrated for the first time that GLP shows prominent anticancer activities against the HCT-116 human colon cancer cell line through triggering intracellular calcium release and the death receptor pathway.

• Journal of Korean Ophthalmic Optics Society
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• v.7 no.2
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• pp.189-195
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• 2002

The aim of this study was to investigate the action of benzalkonium chloride (BAC) used as a preservative in most ophthalmic topical solutions, on human corneal epithelial (HCE) cells in vitro. HCE cell line was exposed to BAC solutions at various concentrations (0.01%~0.0001%) for 15 minutes followed by 24 hours of cell recovery. Cell viability was assessed using MTT assay and chromatin condensation with a Hoechst 33342 test. The expression of membrane protein Fas and Fas ligand was examined by western blot and immunocytochemistry, and DNA fragmentation was studied by agarose gel electrophoresis. A significant decrease of membrane integrity with chromatin condensation was observed with BAC tested at concentrations of 0.005% and higher. BAC was cytotoxic preservatives in this study. An apoptotic mechanism appeared to be present at low concentrations of BAC, whereas a necrotic process appeared at higher concentrations. A functional Fas-mediated apoptotic pathway is present in cultured HCE cells and can be activated by upregulation of Fas expression with BAC.