• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.539-542
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• 2001

$^1H$-NMR spectroscopy was used to detect apoptosis in HL-60 cells in vitro. The relationship between cell apoptosis and NMR data was validated by the flow cytometry assay. To evaluate the NMR apoptosis results, the ratio of methylene and methyl groups caused by lipids was used. In addition, an identical analysis was applied to HepG2 cells. Detection of apoptotic cell death by NMR spectroscopy was oserved.

• Journal of Applied Biological Chemistry
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• v.63 no.3
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• pp.197-201
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• 2020

In order to enhance the extraction efficiency of flavonoid from Saururus chinensis, carbohydrate-hydrolyzing enzyme Viscozyme L aided extraction techniques have been studied. Then flavonoid composition, as well as quercetin, were also identified using UV/Vis, HPLC/MS, and ¹H-NMR. The results showed that favorable extraction conditions were Viscozyme L concentration of 0.25 mg/g, pH 4.2, reaction at 45 ℃ for 12 h. Under the favorable extraction condition, total flavonoid yield (37.9 mg/g) and quercetin yield (0.86 mg/g) increased by about 2.0 and 9.6 times, respectively, compared to control group. Interestingly, as a significant flavonoid of S. chinensis, flavonoid glycones rutin was hydrolyzed to aglycones quercetin by Viscozyme L. These findings provide scientific and theoretical support for the development quercetin-rich products, which was quickly absorbed by the human body than rutin.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.329-333
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• 2003

Human granulocyte-macrophage colony-stimulating factor (hGM-CSF) is a glycoprotein that stimulates the production of granulocytes, macrophages and white blood cells. hGM-CSF secreted by transgenic Nicotiana tabacum suspension cells was unstable in the culture medium and rapidly degraded by extracellular preteases. In order to reduce extracellular pretense activity, culture temperature was lowered. Then, the production of hGM-CSF by transgenic plant suspension cell cultures could be enhanced by reduced degradation of hGM-CSF at low temperature.

• Journal of Life Science
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• v.10 no.2
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• pp.210-217
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• 2000

A kinetic profile of the catalytic domain of stromelysin-1 (SCD) using the fluorescent peptide substrate has been determined by the stopped-flow technique. The pH profile has a pH optimum of about 5.5 with an extended shoulder above pH 7. Three pKa values, 5.0, 5.7, and 9.8 are found for the free enzyme state and two pH independent Kcat/Km values of 4.1$\times$104 M-1 s-1 and 1.4$\times$104 M-1 s-1 at low and high pH, respectively. The profile is quite different in shape with other MMP family which has been reported, having broad pH optimum with two pKa values. The substrate specificity of SCD towards fluorescent heptapeptide substrates has been also examined by thin layer chromatography. The cleavage sites of the substrates have been identified using reverse-phase HPLC method.SCD cleaves Dns-PLA↓L↓WAR and Dns-PLA↓L↓FAR at two positions. However, the Dns-PLA↓LRAR, Dns-PLE↓LFAR, adn Dns-PLSar↓LFAR are cleaved exclusively at one bond. The double cleavages of Dns-PLALWAR and Dns-PLALFAR by SCD are in marked contrast to the close structurally related matrilysin. A notable feature of SCD catalysis agrees with the structural data that the S1' pocket of SCD is deeper than that of matriysin. The differences observed between SCD and matrilysin may form the basis of understanding the structural relationships and substrate specificities of the MMP family in vivo.

• The Journal of Korean Medicine
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• v.27 no.4
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• pp.162-173
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• 2006

The water-extracts of Scutellaria barbata Don (SBDE) were isolated from Chinese medicinal plant sources. The extracts showed strong growth-inhibitory activity and cancer chemopreventive activity on the growth and DNA incorporation of MG63 human osteosarcoma and K562 human leukemia cell lines. The growth of human cancer cells was inhibited in the presence of the extracts (20, 50 and 100 ${\mu}$g/ml), and the effects were concentration-dependent and incubation time-dependent up to 8 days. When 50 ${\mu}$g/ml of the extracts was added to the media of MG63 and K562, cell growth after 8 days or 6 days of incubation was retarded by 93.2 to 97.3% of the control group. Morphological changes of MG63 and K562 cell lines were observed. As the concentration of the extracts increased up to 50 ${\mu}$g/ml, degree of cell aggregation decreased. Moreover, the DNA incorporation of the cells which were labeled with [3H] thymidine was significantly reduced after 3 days of incubation at $37^{\circ}C$ with the extract. Therefore, it is suggested that the extract is highly effective on inhibition of cancer cell growth. The extract also inhibited gene expression of IGF-II in transcriptional level. Since IGF-II works as a mitogenic effector on MG63 and K562 cell lines, these results suggest that the growth inhibition is in part mediated through the inhibition of IGF-II gene expression.

• Journal of Ginseng Research
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• v.44 no.2
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• pp.258-266
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• 2020

Background: Oxidative stress-induced cardiomyocytes apoptosis is a key pathological process in ischemic heart disease. Glutathione reductase (GR) reduces glutathione disulfide to glutathione (GSH) to alleviate oxidative stress. Ginsenoside Rb1 (GRb1) prevents the apoptosis of cardiomyocytes; however, the role of GR in this process is unclear. Therefore, the effects of GRb1 on GR were investigated in this study. Methods: The antiapoptotic effects of GRb1 were evaluated in H9C2 cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, annexin V/propidium iodide staining, and Western blotting. The antioxidative effects were measured by a reactive oxygen species assay, and GSH levels and GR activity were examined in the presence and absence of the GR inhibitor 1,3-bis-(2-chloroethyl)-1-nitrosourea. Molecular docking and molecular dynamics simulations were used to investigate the binding of GRb1 to GR. The direct influence of GRb1 on GR was confirmed by recombinant human GR protein. Results: GRb1 pretreatment caused dose-dependent inhibition of tert-butyl hydroperoxide-induced cell apoptosis, at a level comparable to that of the positive control N-acetyl-L-cysteine. The binding energy between GRb1 and GR was positive (-6.426 kcal/mol), and the binding was stable. GRb1 significantl reduced reactive oxygen species production and increased GSH level and GR activity without altering GR protein expression in H9C2 cells. Moreover, GRb1 enhanced the recombinant human GR protein activity in vitro, with a half-maximal effective concentration of ≈2.317 μM. Conversely, 1,3-bis-(2-chloroethyl)-1-nitrosourea co-treatment significantly abolished the GRb1's apoptotic and antioxidative effects of GRb1 in H9C2 cells. Conclusion: GRb1 is a potential natural GR agonist that protects against oxidative stress-induced apoptosis of H9C2 cells.

• Biomedical Science Letters
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• v.27 no.4
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• pp.277-282
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• 2021

Hydrocotyle is a genus of prostrate, perennial aquatic or semi-aquatic plants formerly classified in the family Apiaceae, now in the family Araliaceae. Lipoxygenases (LOX) are present in the human body and play an important role in the stimulation of inflammatory reactions. Ethanolic extracts of five Hydrocotyle species (H. ramiflora, H. maritima, H. nepalensis, H. sibthorpioides, and H. yabei) showed inhibition of 23.5~50.6% at 2.0 mg/mL. Their extracts showed LOX inhibition in half maximal effective concentration (EC₅₀) range 15.1~15.7 ㎍/mL. Hyaluronic acid is a glycosaminoglycan, a major component of the extracellular matrix Five extracts of these species inhibited less than 23.0% of Hyaluronidase (HAase) activity at a concentration of 2.0 mg/mL Xanthine oxidase (XO) is a form of xanthine oxidoreductase, a type of enzyme that generates reactive oxygen species. Five Hydrocotyle species were found to have inhibitory activity of XO at 2.0 mg/ml, with 65% having greater than 50% inhibition. H. ramiflora exhibited the highest activity with an inhibition of 80.0%. The results suggested that Lipoxygenases, Hyaluronidase, and Busan 47340, Republic of Korea from five Hydrocotyle species might be multifunctional and prevent the degradation of allergic reactions and inflammation.

• Journal of Life Science
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• v.23 no.1
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• pp.84-94
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• 2013

The cytotoxicity of coffee-like green tea (CLGT) was determined in a human breast cancer cell line, MCF-7; a human prostate cancer cell clone, PC-3; a human neuroblastoma cell line, SK-N-SH; and a rat cardiomyoblast cell line, H9c2, with reference to green tea leaves (GTL). The CLGT was prepared by roasting the GTL for 60 min at $240^{\circ}C$ in a temperature-controlled frying pan. The CLGT preparation imitated the flavor and taste characteristics of coffee fairly well according to sensory analysis. The CLGT preparation had no adverse cytotoxic effects on the cancer cells or the normal cells compared to GTL. No significant change in the antioxidant activity was seen in the CLGT preparation compared to that of GTL. The amount of total protein, sugar, and phenolic compounds was reduced in the preparation relative to those in GTL, a fact that might explain the coffee-like flavor and/or taste characteristics of the CLGT preparation. These results suggest that CLGT prepared by roasting GTL for 60 min at $240^{\circ}C$ does not show any adverse effects on cancer cells and normal cells compared to GTL. They imply that CLGT could be safe for human consumption.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.1
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• pp.35-42
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• 2019

We investigated the protective effects of antioxidant fractions from a 70% ethanolic extract of Hizikia fusiformis in human dermal fibroblasts (HDFs). Powdered H. fusiformis was extracted with 70% ethanol and then partitioned into three fractions according to polarity using n-hexane (HFH), chloroform (HFC), and ethyl acetate (HFEA). Antioxidant activity was observed in HFEA at 0.66 mg/mL based on the half maximal inhibitory concentration ($IC_{50}$) of 1,1-diphenyl-2-picrylhydrazyl (DPPH), and at 0.24 mg/mL based on alkyl radical scavenging. The protective effects of the HFEA antioxidant fraction against 2,2-azobis-(2-amidinopropane) dihydrochloride (AAPH)-damaged HDFs and the expression of Type I procollagen in HDFs were examined. HFEA caused the proliferation of HDFs with and without AAPH treatment and protected against AAPH damage to HDFs in a dose-dependent manner ($50-200{\mu}g/mL$). This implies that the antioxidant properties of the fractions depended on their proliferative and protective effects. The HFEA antioxidant fraction had significant effects and caused the dose-dependent expression of Type I procollagen, an important anti-wrinkle protein, in HDFs. In conclusion, antioxidant substances in H. fusiformis were found in the ethyl acetate fraction, and the resulting HFEA may have cosmetic applications.

• Oncology Reports
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• v.43 no.1
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• pp.358-367
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• 2020

The thioredoxin (Trx) system is an important enzymatic complex involved in cellular redox homeostasis. Arsenic trioxide (ATO; As2O3) is known to trigger cell death in vascular smooth muscle cells (VSMCs) via oxidative stress. In the present study, the effects of changes in thioredoxin 1 (Trx1) and Trx reductase1 (TrxR1) on cell growth, death, reactive oxygen species (ROS), and glutathione (GSH) levels were evaluated in ATO-treated human pulmonary artery smooth muscle cells (HPASMCs). ATO inhibited growth and induced cell death in the HPASMCs at 24 h. Overexpression of Trx1 and TrxR1 using adenoviruses attenuated cell growth inhibition caused by ATO and partially prevented cell death. ATO increased ROS levels including the mitochondrial superoxide anion (O2•-) at 5 min. Administration of adTrx1 or adTrxR1 reduced the increased mitochondrial O2•- level in these cells. HPASMCs treated with Trx1 or TrxR1 siRNA showed increases in ROS levels with or without treatment of ATO at 5 min. Although ATO transiently increased GSH levels at 5 min, Trx1 and TrxR1 siRNAs reduced the increased GSH levels in these cells. In addition, PX-12 (a Trx1 inhibitor) and auranofin (a TrxR1 inhibitor) diminished the cellular metabolism in HPASMCs at 4 h, accompanied by an increase in ROS level and a decrease in GSH level. In conclusion, upregulation of Trx1 and TrxR1 somewhat decreased cell growth inhibition and death in ATO-treated HPASMCs, which was accompanied by reduced oxidative stress.