• The Korea Journal of Herbology
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• v.22 no.2
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• pp.155-161
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• 2007

Objectives: Evidences suggests that Ginkgo biloba, a widely used traditional medicine, shows a hypoglycemic effect. Thus, we investigatd the effect of G. biloba extract (GB) on glucose uptake in L6 rat skeletal muscle cells. Method : Effect of GB on glucose uptake and phosphatidylinositol (PI) 3-kinase activity were assessed using Glucose uptake assay and PI 3-kinase assay, respectively. Also, AMP-activated protein kinase (AMPK), p38 mitogen activated protein kinase (p38 MAPK) expression were identified by Western blot. Results : Glucose uptake assay revealed that GB increased glucose uptake about 2.5-fold compared to thecontrol. GB stimulated the activity of PI 3-kinase which is a major switch element on the glucose uptake pathway. About a 6.5-fold increase in activity of PI 3-kinase was observed with GB. We then assessed the activity of AMPK, another regulatory molecule on the glucose uptake pathway. The result was that GB increased the phosphorylation level of both AMPK ${\alpha}$l and ${\alpha}$2. The activity of p38 MAPK, a downstream mediator of AMPK, was also increased by CB. Conclusion : These results suggest that GB may stimulate glucose uptake through both PI 3-kinase and AMPK mediated pathways in L6 skeletal muscle cells thereby contributing to glucose homeostasis.

• Journal of Pharmacopuncture
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• v.16 no.2
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• pp.46-54
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• 2013

Objective: This study was undertaken to isolate a fibrin(ogen)olytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate the enzymatic characteristics and hemorrhagic activity of the isolated enzyme as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were determined by using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrin(ogen)olytic enzyme with the molecular weight of 27 kDa (FE-27kDa) isolated from G. b. siniticus venom consisted of two heterogenous disulfide bond-linked polypeptides with the molecular weights of 15 kDa and 18 kDa. When more than $20{\mu}g$ of FE-27kDa was applied on the fibrin plate, fibrinolysis zone was formed as indicating its fibrinolytic activity. The fibrinolytic activity was inhibited completely by phenylmethanesulfonylfluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA) and partially by thiothreitol and cysteine. Metal ions such as $Hg^{2+}$ and $Fe^{2+}$ inhibited the fibrinolytic activity completely, but $Mn^{2+}$ did not. FE-27kDa preferentially hydrolyzed ${\alpha}$-chain of fibrinogen and slowly hydrolyzed ${\beta}$-chain, but did not hydrolyze ${\gamma}$-chain. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into polypeptides with molecular weights of more than 45 kDa. A dosage of more than $10{\mu}g$ of FE-27kDa per mouse was required to induce hemorrhage beneath the skin. Conclusion: FE-27kDa was a serine proteinase consisting of two heterogeneous polypeptides, hydrolyzed fibrin, fibrinogen, and gelatin, and caused hemorrhage beneath the skin of mouse. This study suggests that the potential of FE-27kDa as pharmacopuncture agent should be limited due to low fibrinolytic activity and a possible side effect of hemorrhage.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.4
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• pp.133-137
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• 2011

The monitoring of heparin therapy is using almost aPTT assay. This study is compare to estimating aPTT therapeutic range using in vitro heparin-spiked sample and aPTT therapeutic range using in vivo heparin-treated sample. Normal pooled plasma was collected from 20 healthy representative individuals. 11 concentration of heparinized plasmas from 0 U/mL to 1.0 U/mL at intervals of 0.1 U/mL made by addition of heparin to normal pooled plasma were measured aPTT. The aPTT therapeutic range was performed through correlation analysis between heparin level 0.2 to 0.4 U/mL and aPTT. 30 plasmas from patients on heparin therapy were measured aPTT and anti-Xa activity. The aPTT therapeutic range was performed through correlation analysis between anti-Xa activity 0.3 to 0.7 U/mL and aPTT. The aPTT therapeutic range corresponded by heparin level-vs-aPTT value regression analysis was 60.7 to 102.4 seconds. The aPTT therapeutic range corresponded by anti-Xa activity-vs-aPTT value regression analysis was 85.3 to 147.5 seconds. The validation of heparin sensitivity using in-vitro heparin sample was not considered. The establishing aPTT therapeutic range is recommended anti-Xa activity using in-vivo sample.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.12
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• pp.4919-4923
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• 2014

Background: To investigate the effect of celecoxib on telomerase activity and apoptosis in a human laryngeal squamous carcinoma cell line (Hep-2 cells). Materials and Methods: The growth inhibition rate of Hep-2 cells in vitro was measured by MTT assay, and apoptosis by TUNEL assay and flow cytometry (FCM). The TRAP-ELISA method was used to determine telomerase activity in Hep-2 cells. The mRNA expression of human telomerase RNA component(hTR), human telomerase reverse transcriptase (hTERT) and human telomerase-associated protein(hTEP1) was determined by RT-PCR assay. Expression of Bax and Bcl-2 proteins was assessed by Western blotting. Results: Celecoxib can inhibit proliferation and induce apoptosis in a dose- and time-dependent manner, repress telomerase activity, decrease hTERT mRNA and Bcl-2 protein expression and increase Bax protein expression, PGE2 had no effect on telomerase. Conclusions: Celecoxib had the antiproliferative and pro-apoptotic effect in Hep-2 cells. Apoptosis was accompanied by a decrease in telomerase activity which was directly correlated with hTERT mRNA and up-regulation of Bax/Bcl-2. Bcl-2 may thus play an important role in telomerase activity as well as apoptosis.

• Journal of Society of Preventive Korean Medicine
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• v.4 no.1
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• pp.144-151
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• 2000

This study was carried out to evaluate cytotoxic effects of Houttuynia cordata Thunberg extracts on murine leukemia tumor cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazol iumbromide (MMT) assay. The comparison of $IC_{50}$ values of Houttuynia cordata Thunberg extracts on $L1210,\;P388D_1$ and Vero cell lines showed that the methanol extract of Houttuynia cordata Thunberg indicated the most antitumor activity in the MTT assay. In order to develop a antimicrobial agent, dried Houttuynia cordata Thunberg was extracted with several solvents, and then antimicrobial activity was investigated. The minimal inhibitory concentration (MIC) of the extracted substance against microorganisms were also examined. Antimicrobial activity of amocla and ketoconazole as references was compared to those of other solvent extracts such as $H_2O$, n-hexane, chloroform, ethyl acetate ethanol and methanol. The antimicrobial activity of all extracts from the sample had growth inhibition activity against gram-negative bacteria, yam-positive bacteria and fungi $(MIC,\;>\;200\;{\mu} g/ml)$. These results suggest that the methanol soluble extract of Houttuynia cordata Thunberg may be a valuable choice for the studies on the treaeent of murine leukemia tumor cell lines and antimicrobial agents.

• Biomolecules & Therapeutics
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• v.21 no.1
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• pp.72-78
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• 2013

This work aimed to assess some pharmacological activities of P. leptostachya var. asiatica Hara. The dried roots of P. leptostachya var. asiatica Hara were extracted with 70% ethanol to generate the powdered extract, named PLE. Anti-angiogenic activity was detected using chick chorioallantoic membrane (CAM) assay. In vitro anti-inflammatory activity was evaluated via analyzing nitric oxide (NO) content, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Antioxidant activity was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and reactive oxygen species (ROS) level in the stimulated macrophage cells. Matrix metalloproteinase-9 (MMP-9) and -2 (MMP-2) activities in the culture media were detected using zymography. PLE exhibits an anti-angiogenic activity in the CAM assay, and displays an inhibitory action on the generation of NO in the LPS-stimulated macrophage cells. In the stimulated macrophage cells, it is able to diminish the enhanced ROS level. It can potently scavenge the stable DPPH free radical. It suppresses the induction of iNOS and COX-2 and the enhanced MMP-9 activity in the stimulated macrophage cells. Both monooxygenase and oxidase activities of tyrosinase were strongly inhibited by PLE. Taken together, the dried roots of P. leptostachya var. asiatica Hara possess anti-angiogenic, anti-inflammatory, antioxidant and skin whitening activities, which might partly provide its therapeutic efficacy in traditional medicine.

• Journal of Food Hygiene and Safety
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• v.24 no.2
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• pp.124-127
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• 2009

The aim of this study is to evaluate the estrogenic activity of Cucurbita pepo seed extract which includes ${\beta}-sitosterol$ and other phytosterols. Sample was extracted from Cucurbita pepo seed by supercritical carbondioxide method and resuspended with ethanol. Estrogenic activity was measured by recombinant yeast assay which detects estrogenic activity using recombinant yeast with high level of estrogenic receptor. However, estrogenic activity of pumpkin seed extract was not found in this study. Based on this data, pumpkin seed extract will not cause estrogenic disturbance.

• Journal of Life Science
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• v.11 no.5
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• pp.432-438
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• 2001

Various lactic acid bacteria were isolated from Korean Dongchimi (whole radish Kimichi with added water) in order to study their antimutagenic activity. Ames test using Salmonella enterica serovar typhimurium TA98 and TA100 showed the strain DLAB19 to have the highest antimutagenic activity among the 300 isolated strains against MNNG(N-methyl-N-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), 4-NQO(4-nitroquinoline-1-oxide) and AFB$_{1}$(aflatoxin B$_{1}$). The strain was identified as Leuconostoc mesenteroides subsp. cremoris according to the Bergeys Mannual Systematic Bscteriology based on its morphological, cultural, physiological characteristics and biological system Antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was found in the culture supernatant suggesting the bacterium secretes, the antimutagenic substance in the media. The antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17 (Rec$^{+}$) and M45 (Rec$^{[-10]}$ ).).

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.155-162
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• 2008

Objectives : This study was evaluated to elucidate the inhibitory potential of Bodusan (BDS) and its components, Strychnos ignatii semen (SIS) and Glycyrrhizae Radix (GR), on human aortic smooth muscle cells (HASMC) migration and production of matrix metalloproteinase (MMP)-2 and MMP-9 by $TNF-{\alpha}$ treatment. Methods : Cytotoxic activity of BDS and its components on HASMC was using 5-(3-caroboxymeth- oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Effect of BDS, SIS and GR on $TNF-{\alpha}-induced$ HASMC migration underside of matrigel filter was stained with hematoxylin-eosin. And total number of cells that migrated to the underside of the filter was counted. MMP-2 and MMP-9 activity was evaluated by gelatin zymography assay. Results : The matrigel migration assay showed that BDS effectively inhibited the $TNF-{\alpha}-induced$ migration of HASMC. Moreover, BDS significantly inhibited MMP-9 activity. Our present study demonstrates that BDS and its components inhibits $TNF-{\alpha}-induced$ HASMC migration and MMP-9 activity. The inhibitory effect of BDS extract is more potent than that of its component herb extracts. Conclusions : These results provide evidence that BDS has multiple effects in the inhibition of HASMC migration and may offer a therapeutic approach to block HASMC migration.

• Food Science and Preservation
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• v.18 no.6
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• pp.817-823
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• 2011

Angelicae gigas Nakai has been used as a traditional medicine as well as an edible vegetable in South Korea. In this study, the total phenolic and flavonoid content and antioxidants of A. gigas Nakai leaves were examined in vitro via hydroxyl-radical-scavenging activity, reducing power activity, metal chelating assay, and DPPH-free -radical-scavenging assay. Among all the extracts from A. gigas Nakai leaves, the ethanol extract showed the strongest effects in all of the assays. The $EC_{50}$ values for the DPPH-radical-scavenging activities of ethanol, methanol, and water extracts were 31.47, 42.14, and $58.47{\mu}g/mL$, respectively. Among the extracts from A. gigas Kakai leaves, the ethanol extract had the highest levels of total phenolics ($7.84{\pm}1.46$ mg TAN/g) and total flavonoids ($4.23{\pm}0.03$ mg QE/g), which correlated strongly with the individual phenolic-compound (p-hydroxybenzoic acid, vanillin, and trans-ferulic acid) contents. The ethanol extract also showed stronger antioxidant activity than tocopherol in hydroxyl- radical-scavenging activity assay. These results indicate that the ethanol extract of A. gigas Kakai leaves possesses significant antioxidant properties, which suggests its great potential as a functional-food ingredient in the food industry.