• Journal of Oriental Neuropsychiatry
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• v.18 no.2
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• pp.13-24
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• 2007

Objective : Recent studies indicate that the deposition of ${\beta}-amyloid$ ($A{\beta}$) is related in the pathogenesis of Alzheimer's disease (AD), but the underlying mechanism is still not clear. Method : To investigate the potential cellular functions of APP and water extract of the JangwonHwangagambang (JWHG), we use as in vitro model, neuro 2A cells were treated with either JWHG or its oriental medicines, and the effect in APP expression was determined by MTT and LDH assay. JWHG have been shown to be neuroprotective in different model systems. We asked whether JWHG treatment would influence cell survival and AD-like pathology in APP-induced neuronal cells. Result : JWHG and water extracts of some oriental medicine has attenuated high cell death in vitro. JWHG-treated cells increased percentage of cell survival more longly than controls. JWHG had significantly increas neurite outgrowth in the as compared to control cells. Conclusion : These results suggest that JWHG prevent APP-induced neurotoxicity through attenuating oxidative stress, and may be useful as potential therapeutic agents for AD.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.4
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• pp.674-680
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• 2010

The purpose of this study was to investigate the effects of Fructus Ligustri Lucidi (FLL) on the process of melanogenesis. Cell viability of B16F10 cells was measured by MTT assay and melanin content was assessed using the method of Hosoi with some modifications. Methanol extract of Fructus Ligustri Lucidi (MFLL) significantly inhibited melanin synthesis in a concentration-dependent manner, and it reduced the activity of tyrosinase, the rate-limiting melanogenic enzyme. Additionally, $\alpha$-melanocyte stimulation hormone ($\alpha$-MSH)-induced hyperpigmentation was down-regulated by MFLL. MFLL was fractionated by organic solvent. In the present study, Hexane extract of Fructus Ligustri Lucidi (HFLL) inhibited tyrosinase activity and melanin production of B16F10 cells in the absence or presence of $\alpha$-MSH. Ethyl acetate, butanol and water layers did not affect tyrosinase activity and melanin production. Meanwhile ethyl acetate, butanol and water layers showed DPPH free radical scavenging activity. These results suggest that extract of FLL could be used as functional biomaterial in developing a skin whitening agent having the antioxidant activity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.1
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• pp.111-117
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• 2010

The purpose of this study was to investigate the effect of garlic(Allium sativum L.) extracts with extraction conditions on antioxidant and anticancer activity. The extracts prepared for garlic by hot temperature extraction (HG), low temperature extraction (LG), UMPM extraction (UG), fermentation (FG) and black garlic hot temperature (BG) method. Content of total polyphenol compound was the BG higher than other extracts. The EDA (electron donating ability) and SOD-like activity was increased in dose-dependent manners, and the activity of BG and UM was significantly higher than LG and FG. We examined cytotoxicity, nitric oxide production of Raw 264.7 cell and inhibition of HT 1080 cell by MTT assay. All extracts does not have any toxic effects in macrophages(Raw 264.7). And UG inhibited the production of nitrite in Raw 264.7 cells activated with LPS. The antitumor effects of LG and UG on HT 1080 cell was indicated a significantly inhibition activity. These results suggested that UG (UMPM extraction of garlic) have activities of antioxidant, anticancer effects.

• Archives of Plastic Surgery
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• v.33 no.1
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• pp.5-12
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• 2006

Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, take a significant role in signal transduction pathway of angiogenesis. The authors utilized the inhibitors, targeting the formation of three co-enzyme in signal transduction pathway in order to quantify the suppression of abnormal vascular endothelial cell proliferation induced by DMH, to compare the level suppression in each up-regulated growth factors, CTGF, CYR61, $ITG{\beta}1$, FHL2, and to identify the relationship between abnormal cell proliferation and signal transduction pathway. Five groups were established; Control group, Group of DMH, Group of DMH-mixed Herbimycin, inhibitor of protein tyrosine kinase, Group of DMH-mixed Calphostin C, inhibitor of protein kinase C, Group Of Dmh-Mixed 10U Catalase, Inhibitor Of oxidase. The rise of vascular endothelial cell was compared by MTT assay, and four growth factors were analysed with RT-PCR method, at pre-administration, 4, 8, 12, and 24 hours after administration. In comparison of abnormal proliferation of vascular endothelial cell induced by DMH, suppression was noticed in Herbimycin and Calphostin C group, and Calphostin C group revealed higher suppression effect. Nevertheless, Catalase group did not have any suppression. In manifestation of four growth factors, Herbimycin and Calphostin C group presented similar manifestation with control group, except in $ITG{\beta}$. Catalse group had similar manifestation with DMH group in all four growth factors. Abnormal proliferation of vascular endothelial cell induced by DMH have a direct relationship with PTK and PKC, more specifically to PKC. Oxidase was confirmed not to have any relevance.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.4
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• pp.603-607
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• 2011

The purpose of this study is to investigate the effects of Carthami Flos on interstitial cells of Cajal in the gastrointestinal tract. Many regions of the tunica muscularis of the gastrointestinal (GI) tract display spontaneous contraction. These spontaneous contractions are mediated by periodic generation of electrical slow waves. Recent studies have shown that the interstitial cells of Cajal (ICCs) act as pacemakers and conductors of electrical slow waves in gastrointestinal smooth muscles. We investigated the cytotoxicity activity, antioxidant activity, and pacemaking activity. The cytotoxicity activity was measured by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Antioxidant activities were determined by DPPH (1.1-diphenyl-2-picrylhydrazyl) radical scavenging capacity assay and DCFH-DA (2,7-dichlorofluorescein diacetate) method. The effects of Carthami Flos on the pacemaker potentials in cultured ICCs from murine small intestine were investigated by using whole-cell patch-clamp techniques at $30^{\circ}C$. The addition of Carthami Flos (5, 10, $30{\mu}g$/ml) depolarized the resting membrane potentials in a concentration dependent manner. These results suggest that the GI tract can be targets for Carthami Flos, and their interaction can affect intestinal motility.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.6
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• pp.956-961
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• 2010

Tumor necrosis factor-alpha (TNF-${\alpha}$) is a major mediator of immuno-inflammatory activity. The purpose of this study is to investigate whether TNF-${\alpha}$ productions of mouse macrophage RAW 264.7 and human hepatocyte HepG2 are modulated by Artemisiae argi Folium water extract (AW), Lactobacillus pentosus-fermented Artemisiae argi Folium water extract (AFL), and Saccharomyces cerevisiae-fermented Artemisiae argi Folium water extract (AFS) for 3 h of incubation. Effect of AW on cell viability of HepG2 was also investigated. TNF-${\alpha}$ productions were measured by Enzyme-Linked Immnunosorbent Assay method and cell viability was measured by MTT assay. Both AFL and AFS significantly increased TNF-${\alpha}$ productions of RAW 264.7 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). Also, AFL and AFS significantly increased TNF-${\alpha}$ productions of HepG2 at the concentration of 50, 100, and 200 ${\mu}g$/mL (p<0.05). AW significantly increased TNF-${\alpha}$ production of HepG2 at the concentration of 100 and 200 ${\mu}g$/mL (p<0.05). AW did not show any cytotoxicity on HepG2 cells for 3 h. These results suggest that AFL, AFS, and AW have the immune-enhancing property related with its increasing effect on TNF-${\alpha}$ production of macrophage and hepatocyte.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.159-160
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• 1998

Results of chemistry and biological activity of many years indicate that plants belonging to the Isodon genus are rich in ent-kaurane diterpenoids, which have been revealed to possess biological activities such as antitumor, antibacterial and antiinflammatory effects. In continuation of our research on diterpenoids in medicinal plants of this genus, the acetone extract from the leaves of I. xerophilus, which is a plant native to Yunnan province of China, showed potent antitumor activity against K562. After partition, the most active EtOAc part was studied. Four new diterpenoids named xerophilusin A(l), B(2), C(3), D(4), and eight known compounds including macrocalin B(5) and rabdorosthomin A(6) were isolated, whose structures were elucidated through a series of one- and two-dimensional NMR techniques(DEPT, COSY, HMQC, HMBC and ROESY experiments). Among them, compound 1, 2 and 5 had two unique epoxy units formed by two ether bridges from C-20 to C-7, C-14. Up to now, there are four compounds having such an peculiar structure besides these three compounds. Compound 3 and 4 were two of the few examples possessing $1{\beta}$ substitutes. All the diterpenoid compounds were subjected to the antitumor screening. It is interesting that only xerophilusin A(l), B(2) and macrocalin(5) exhibited significant antitumor activity against K562 by the method of MTT($IC_{50}$ were listed in Table 1.). The results inspired us to infer that the unique ether bridges from C-20 to C-7, C-14 possibly played an important role in the antitumor activity.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.577-582
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• 1997

Effects of four citrus flavonoids, hesperidin, naringin and their aglycones on phosphatidate phosphohydrolase(PAP, EC 3.1.3.3) activity were examined using isolated rat microsomes as an enzyme source. In addition, these flavonoids were tested to see whether they exert any influence on the proliferation and growth in cultured human hepatocytes HepG2 cells. Flavonoids at concentration up to $10{-4}M$ had no significant effect on the number of cells and cell proliferation by MTT cell growth assay method, whereas aglycone flavonoids, hesperetin and narigenin, at concentration of $10{-3}M$ significantly inhibited cell proliferation. Hesperetin inhibited PAP activity in a dose-dependent manner starting at concentration of $10{-5}M$. Narigenin at concentration of $10{-2}M$ inhibited PAP activity markedly, while the other flavonoids did not show any significant effect. The present study, therefore, demonstrated that aglycone flavonoids exerted portent effects on PAP activity and on cell proliferation.

• Tuberculosis and Respiratory Diseases
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• v.72 no.4
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• pp.343-351
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• 2012

Background: The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling axis has emerged as a novel target for cancer therapy. Agents that inhibit this pathway are currently under development for lung cancer treatment. In the present study, we have tested whether dual inhibition of PI3K/Akt/mTOR signaling can lead to enahnced antitumor effects. We have also examined the role of autophagy during this process. Methods: We analyzed the combination effect of the mTOR inhibitor, temsirolimus, and the Akt inhibitor, GSK690693, on the survival of NCI-H460 and A549 non-small cell lung cancer cells. Cell proliferation was determined by MTT assay and apoptosis induction was evaluated by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Autophagy induction was also evaluated by acridine orange staining. Changes of apoptosis or autophagy-related proteins were evaluated by western blot analysis. Results: Combination treatment with temsirolimus and GSK690693 caused synergistically increased cell death in NCI-H460 and A549 cells. This was attributable to increased induction of apoptosis. Caspase 3 activation and poly(ADP-ribose) polymerase cleavage accompanied these findings. Autophagy also increased and inhibition of autophagy resulted in increased cell death, suggesting its cytoprotective role during this process. Conclusion: Taken together, our results suggest that the combination of temsirolimus and GSK690693 could be a novel strategy for lung cancer therapy. Inhibition of autophagy could also be a promising method of enhancing the combination effect of these drugs.

• YAKHAK HOEJI
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• v.55 no.2
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• pp.121-126
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• 2011

Sipjundaebo-tang was a well-known restorative traditional herbal prescription that used to treat anemia, anorexia, fatigue and inflammation. In this study, we examined the bioconversion of compounds in the Sipjundaebo-tang (SJ) and fermented Sipjundaebo-tang with Lactobacillus fermentum KFRI 164 (FSJ) using established HPLC-DAD method. The chromatogram of FSJ has shown that the contents of six bioactive compounds 5-HMF, paeoniflorin, ferulic acid, cinnam aldehyde, decursin, glycyrrhizin in SJ has decreased. And the contents of unknown compounds (1), (2), (3), (4) and (5) in FSJ were higher than each contents of SJ. The antioxidant activities of SJ and FSJ were conducted by DPPH free radical and Hydrogen peroxide ($H_2O_2$) scavenging assay. Electron donating activity (EDA, %) value of FSJ has shown higher than 21.9% and 14.5% at a concentration of 0.5 mg/ml for DPPH radical scavenging activity and $H_2O_2$ scavenging activity, respectively. Also, the neuroprotective activities of SJ and FSJ against glutamate-induced oxidative stress in a mouse hippocampal cell line (HT22) were evaluated by MTT assay. As a result, FSJ has shown higher neuroprotective activity than 56.5% comparing with SJ. In conclusion, the fermented SJ using microorganism could convert compounds in SJ and enhance antioxidant activity and neuroprotective activity.