• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.5
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• pp.893-898
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• 2002

Citri Reticulatae Viride Pericarpium extract(CRVP) have been used in oriental medicine for many centuries as a therapeutic agent for smoothing the liver and regulating the circulation of qi, and promoting digestion and removing stagnated food. The effects of CRVP on the inhibition of brain damage in cerebral ischemia is not known. Therefore, this Study was designed to investigate the cerebral protective effects of CRVP on the transient cerebral ischemia using modern techniques, and further to provide the possibility of scientification of oriental medicine. The size of cerebral infarct size was measured by morphometry, and brain edema was measured by morphometry and brain water content determination. The results were a$ follows ; 1. Water fraction of CRVP was reduced infect area of rats brain slices which were subjected to a transient cerebral ischemia in a dose-dependent manner. 2. Methylene chloride fraction and hexane fraction of CRVP was significantly reduced infarct area of rats brain slices which were subjected to a transient cerebral ischemia in a dose-dependent manner. 3. Methylene chloride fraction and hexane fraction of CRVP was significantly reduced infarct volume of rats brain which was subjected to a transient cerebral ischemia in a dose-dependent manner. 4. Methylene Chloride fraction and hexane fraction of CRVP was significantly decreased brain edema induced by a transient cerebral ischemia in a dose-dependent manner. 5. Methylene chloride fraction and hexane fraction of CRVP was significantly decreased brain water content of rats which were subjected to a transient cerebral ischemia. It is suggested that CRVP has an anti-ischemic effect through the inhibition of brain damage in a transient cerebral ischemia, and that in future further development of main effective constituent in CRVP can provide a novel therapeutic strategy for cerebral ischemia.

• The Korean Journal of Food And Nutrition
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• v.28 no.5
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• pp.866-870
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• 2015

In this study, coffee waste was extracted with different solvents such as ethyl acetate, methylene chloride and methanol to investigate the total polyphenol contents, electron donating ability and the inhibitory effect on glutathione S-transferase. The total polyphenol contents were $3,060.61{\pm}357.12{\mu}g\;GAE/mL$ in ethyl acetate, $909.09{\pm}35.71{\mu}g\;GAE/mL$ in methylene chloride, and $1,602.27{\pm}30.36{\mu}g\;GAE/mL$ in methanol. The total polyphenol contents showed a significant difference (p<0.05) between the solvents. The electron donating ability was $80.20{\pm}1.45%$ for ethyl acetate, $81.94{\pm}0.45%$ for methylene chloride, and $85.14{\pm}1.53%$ for methanol. The electron donating abilities were significantly different (p<0.05) between the solvents. The inhibitory effect of the various extracts on glutathione S-transferase (% inhibition) was $92.12{\pm}0.56%$, $88.48{\pm}0245%$ with methylene chloride extract, and $90.85{\pm}0.14%$ with methanol extract. These too were significant different (p<0.05) between the solvents. The two portions of coffee waste extracts obtained from ethyl acetate and methanol showed meaningful results on the total polyphenol contents, and the inhibition effects on glutathione S-transferase. Therefore, they can be utilized to develop health care foods and can be applied as antioxidants for cosmeceuticals.

• The Korea Journal of Herbology
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• v.24 no.4
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• pp.173-179
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• 2009

Objectives : In this study, the effects of different fractions isolated from modified Boyanghwanotang(mBHT) extract on LPS-induced inflammation in BV2 microglial cells were investigated. Methods : mBHT was extracted with water, and then fractionated with n-hexane, methylene chloride, ethylacetate and n-butanol. BV2 cells, a mouse microglia line were incubated with different concentrations of each fraction of mBHT for 30 min, and then stimulated with LPS for 24 h. Cell toxicity was determined by MTT assay. The concentration of nitric oxide (NO) was measured in culture medium by Griess reagent assay. The expression of inducible nitric oxide synthease (iNOS) protein was determined by Western blot. Results : Four fractions of mBHT were significantly inhibited LPS-induced NO productions in BV2 cells in a dose-dependent manner. The methylene chloride fraction of mBHT was most strongly inhibited the NO production compared with those of the others. The methylene chloride fraction of mBHT was also suppressed LPS-induced iNOS expression comparison of other fractions at same concentration ($50\;{\mu}g/ml$) in BV2 cells. Conclusions : The results showed that the methylene chloride fraction of mBHT may have an strong anti-inflammatory property through the inhibition of NO production and iNOS expression in activated microglia, and could a therapeutic potential for the treatment of various brain inflammatory diseases.

• Preventive Nutrition and Food Science
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• v.10 no.4
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• pp.316-325
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• 2005

The antioxidant potential of a $75\%$ methanolic extract of highbush blueberry (Vaccinium corymbosum L.) and its different fractions was investigated using different reactive oxygen species (ROS), nitric oxide (NO.), metal chelating and lipid peroxidation assays. Methylene chloride and $75\%$ methanol fractions showed equally high activities $(IC_{50} 0.010 mg/mL)$ for hydroxyl radical (HO) scavenging. Higher hydrogen peroxide $(H_2O_2)$ scavenging values were reported for the ethyl acetate and methylene chloride fractions and their $IC_{50}$ values were 0.20 and 0.15 mg/mL, respectively. Nitric oxide (NO.) and 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging activities were higher in ethyl acetate and methylene chloride fractions. Chloroform and water fractions showed higher activities in superoxide $(O_2.)$ scavenging. All fractions showed strong metal chelating capacities compared with the commercial antioxidants tested. The $0.1\%$ ethyl acetate fraction showed notable capacity to suppress lipid peroxidation in both fish oil and linoleic acid. Phenolic content was measured in all the fractions and methanolic extract. Among the fractions, ethyl acetate fraction showed the highest phenolic content.

• The Korean Journal of Food And Nutrition
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• v.31 no.6
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• pp.775-782
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• 2018

The purpose of this study was to investigate effects of Crataegus pinnatifida var. psilosa extracts obtained from 95% methanol and water. Methylene chloride, ethyl acetate, and methanol were used to fractionate the 95% methanol extract and each fraction was testified total polyphenol contents, electron donating abilities, the scavenging abilities of superoxide anion radical, as well as hydrogen peroxide. Extraction yields of 95% methanol and water from Crataegus pinnatifida var. psilosa were 25.40%, 23.12% respectively. Total polyphenol contents were $28,708.0{\pm}1,755.05{\mu}g\;GAE/mL$ in 95% methanol, revealing the highest among them, $12,726.67{\pm}479.33{\mu}g\;GAE/mL$ in water extract, $15,854.67{\pm}498.38{\mu}g\;GAE/mL$ in methanol fraction, $11,810.67{\pm}584.48{\mu}g\;GAE/mL$ in ethyl acetate fraction, and $5294.67{\pm}190.36{\mu}g\;GAE/mL$ in methylene chloride fraction. Total polyphenol contents revealed significant differences (p<0.05) between the solvents. In the experiment of the electron donating ability, water extract revealed $84.33{\pm}0.1%$ scavenging ability, the highest. Other extracts and fractions were $81.8{\pm}1.11%$ for water, $79.73{\pm}1.32%$ for ethyl acetate fraction, $75.73{\pm}2.17%$ for methylene chloride fraction, and $42.1{\pm}5.01%$ for methanol fraction, the lowest electron donating ability. Electron donating abilities revealed significant difference (p<0.05) between the solvents. In the experiment of superoxide anion radical scavenging ability, ethyl acetate fraction($0.0026{\pm}0.0002$) had the highest scavenging ability, and the others revealed slight increase rather than decrease in scavenging ability. Hydrogen peroxide scavenging ability revealed the highest in methanol fraction ($-0.00206{\pm}0.00165$) and the others were as follow; water extract ($0.00157{\pm}0.00249$), 95.0% methanol extract ($0.005{\pm}0.0036$), methylene chloride fraction ($0.0039{\pm}0.00364$), and ethyl acetate fraction ($0.0002{\pm}0.00059$).

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.8
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• pp.1284-1288
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• 2005

The methanol extract of Houttuynia cordata repressed the expression of tyrosinase gene of Bl6 mouse melanoma cell containing tyrosinase promoter. The extract repressed expression of tyrosinase gene by about 13$\%$ and 45$\%$ at 10$\mu$g/mL and 100 $\mu$g/mL, respectively. In the MTT assay, the same extract exhibited very low cytotoxicity at 1$\mu$g/mL, 10$\mu$g/mL, and 100 $\mu$g/mL, respectively. The fractions of ethyl acetate, butyl alcohol, and water did not showed the repressive effect on the expression of tyrosinase gone, but methylene chloride fraction layer repressed highly at 10$\mu$g/mL and 100$\mu$g/mL.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.239-240
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• 2003

Type I (collagen) and procollagen are reduced in aged human skin. This reduction could result from increased degration by metalloproteinases and from reduced procollagen synthesis and skin collagenase is required for initiation of the degration of type I collagen. In the present study, we study on assay the collagen and collagenase in natural plants using the fibroblast human skin cell. We select the 15 kind of plants used to herbal and 4 kind of fraction(by methylene chloride, ethyl acetate, n-butanol, water). Among these extract, the ethyl acetate fraction from benincasa hispida/prunus persica, trichosanthes kiriowii, trogopterus xanthipes and methylene chloride fractions from benincasa hispida/prunus persica, torilis japonica and n-butanol fraction from cnidium officinale, chrysanthemum sibiricum were selected for further experiments as they exhibited distinctive amount of collagen compared to other natural extracts. These extracts were again subjected to collagenase assay test. Benincasa hispida/prunus persica extract was shown to have exellent collagen synthesis activity from result of the collagen assay test and the other extract was shown to have over 130% of collagen synthesis activity. But, in the study of collagenase assay test just only trogopterus xanthipes extract was shown to have collagenase inhibition.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.2 s.51
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• pp.153-159
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• 2005

In this study, we investigated the application of an extract from Ephedra sinica which has been composed of traditional Korean medicine as a whitening ingredient. The extract of Ephedra sinica which was obtained from the mixture of methanol and water (1:1) the inhibitory effect of tyrosinase. Then, Ephedra sinica was extracted by two different solvents. One was water and the other was methylene chloride. Only, the water extract of Ephedra sinica showed the inhibitory effort of tyrosinase; the anti-tyrosinase activity with $0.2\%$ of the water extract was $60.6\%$. But the extract of Ephedra sinica in methylene chloride fraction showed little inhibitory effect on tyrosinase. The inhibitory effect of the concentrated water extract of Ephedra sinica was tested on L-DOPA auto-oxidation and melanin synthesis in B-16 melanoma. In L-DOPA auto-oxidation, $0.5\%$ of the concentrated water extract showed $87\%$ of inhibition of L-DOPA auto-oxidation and the $0.75\%$ concentrated Ephedra sinica extract in wafer fraction inhibited $98.8\%$ of that. In melanin synthesis of B-16 melanoma, the concentrated water effect of Ephedra sinica inhibited $70.2\%$ or $79.9\%$ of inhibitory effect on that at the concentration of $0.05\%$ or $0.075\%$, respectively. For verifying the skin whitening effect of the concentrated water extract of Ephedra sinica in vivo, we performed the clinical test of that. The research showed the significant skin whitening effect of a cream containing $0.5\%$ Ephedra sinica extract and the statistical analysis showed a significant difference (p < 0.05) between sample (containing $0.5\%$ Ephedra sinica extract) and placebo after 10 weeks.

• Journal of Nutrition and Health
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• v.31 no.6
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• pp.1076-1081
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• 1998

The purpose of this research was to isolate an active component on immunoytes from 70% MeOH extract of Codonopsis lanceolatae Radix(CLE). CLE was fractionated successively with hexane, methylene chloride, n-butyl alcohol and water, and then the water fraction was separated with molecularporous membrane tubing(m.w. 3,500). Each fraction(50mg/kg) was administered p.o. once a day for 7 days in BALB/c mice respectively. None of these fractions affected the apoptosis and mitochondrial transmembrane potential in thymocyte. Hexane and methylene chloride fractions decreased CD4$^{[-10]}$ CD8$^{+}$ single-positive cells, and the water fraction enhanced CD4$^{+}$ CD8$^{[-10]}$ single-positive cells in thymocyte. The proliferation of thymocytes was decreased by the fraction hexane, but was enhanced by the water fraction. Hexane, methylene chloride and butyl alcohol fractions suppressed the production of nitric oxide, which was not affected by the water fraction. Hexane and butyl alcohol fraction suppressed the phagocytic activity, but water fraction enhanced it. The components(m.w. 3,500 above) separated from the water fraction enhanced the proliferation of thymocyte, the population of CD4$^{+}$ CDB$^{[-10]}$ single-positive cells, and phagocytic activity in macrophage. These results suggest that the stimulative components of proliferation, TH population and phagorytc activity is in the water fraction, and the molecular weight is 3,500 above. (Korean J Nutrition 31(6) : 1076~1081, 1998) 1998)

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.6
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• pp.1622-1628
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• 2005

This study was undertaken to define the effect of SoPung-Tang extract on hypertension in spontaneous hypertensive rat and norepinephrine- induced arterial contraction in rabbit. In order to investigate the effect of SoPung-Tang extract on contracted rabbit carotid arterial strips, transverse strips with intact or damaged endothelium were used for the experiment using organ bath. To analyze the mechanism of SoPung-Tang extract-induced relaxation, SoPung-Tang extract infused into contracted arterial strips induced by norepinephrine after treatment of indomethacin, Nu-nitro-L-arginine, methylene blue or tetraethylammonium chloride. Blood pressure was significantly decreased five days after administration of SoPung-Tang extract. SoPung-Tang extract relax arterial strip with endothelium contracted by norepinephrine, but in the strips without endothelium, SoPung-Tang extract- induced relaxation was significantly inhibited. SoPung-Tang relax arterial strip contracted by norepinephrine, but in the strips contracted by high $K^+$, SoPung-Tang extract-induced relaxation was significantly inhibited. The endothelium-dependent relaxation induced by SoPung-Tang extract was decreased by the pre-treatment of $N{\omega}$-nitro-L-arginine or methylene blue, but it was not observed in the strips pre-treated with indomethacin or tetraethylammonium chloride. When $Ca^{2+}$ was applied, the strips which were contracted by norepinephrine in a $Ca^{2+}$-free solution, arterial contraction was increased. But pre-treatment of SoPung-Tang extract inhibited contractile response to $Ca^{2+}$. We suggest that SoPung-Tang could be applied effectively for hypertension and may suppress influx of extra-cellular $Ca^{2+}$ through the formation of nitric oxide in the vascular endothelial cells.