• The Journal of Korean Medicine
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• v.21 no.3
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• pp.174-185
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• 2000

Objectives : This study was carried out to evaluate the effects of five fractions on cell viability, cell cycle progression and apoptosis. Methods : This study employed MTT assay, Cell cycle analysis, Cpp32 protease assay, DNA fragmentation assay and Quantitative RT-PCR analysis. Results : In MTT assay, the butanol fraction of Injinsaryung-San has showed magnificent viability, while the $H_2O$ fraction and ethylacetate fraction also showed higher viability than the control group. The $H_2O$ fraction of Injinsaryung-San has showed magnificent viability, and butanol fraction and ethylacetate fraction of Injinsaryung-San with etoposide have also showed higher viability than the only etoposide group. Cell cycle analysis showed that each fraction of Injinsaryung-San had no significant effect on the cell cycle. DNA fragmentation assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction carried inhibitory effects on apoptosis induction. Cpp32 protease activity assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction decreased Cpp32 protease activity, with the butanol fraction displaying greater effects. Quantitative RT-PCR showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction suppressed Fas and Bax genes, the butanol fraction increased BcI-2 gene, however no effect on Cpp32. Conclusions : The data shows that the butanol fraction of Injinsaryung-San increases the hepatocyte viability and has the heptocelluar protective effect by the suppression of apoptosis through gene regulation.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.43-43
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• 2023

In this study, we investigated in vitro inhibitory activity of wild-simulated ginseng (WSG) against non-alcoholic fatty liver disease using HepG-2 cells. T0901317 treatment increased the lipid accumulation in HepG-2 cells, but WSG treatment inhibited T0901317-mediated lipid accumulation. In addition, WSG downregulated T0901317-mediated expression of SREBP-1c, ACC, FAS and SCD-1 protein. In addition, WSG increased the phosphorylation level of LKB1 and AMPK. Compound C treatment blocked WSG-mediated downregulation of SREBP-1c protein. In conclusion, WSG is considered to inhibit the accumulation of lipids and triglycerides in HepG-2 cells by inducing the activation of LKB1 and AMPK successively, thereby reducing the expression of FAS, ACC, and SCD-1 through suppression of SREBP-1c expression.

• Journal of Life Science
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• v.29 no.5
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• pp.514-523
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• 2019

We investigated the gene expression patterns and the mechanisms of action of the apoptotic response by microarray analysis of human keratinocyte HaCaT cells treated with ginsenoside Rb1, a saponin of Panax ginseng C. A. Meyer. Genes related to apoptosis, the G2/M transition of the mitotic cell cycle, cell division, mitotic nuclear division, and intracellular protein transport were 2-fold up-regulated in HaCaT cells treated with the ginsenoside Rb1, whereas genes related to DNA repair, regeneration fission, and extracellular matrix organization were 2-fold down-regulated. Apoptosis signaling may be mediated by FAS and PLA2G4A, and pathway analysis indicated that STAT3 might be an upstream regulator of these genes. The activity of FAS and PLA2G4A was verified by qPCR, which showed that FAS was increased about 2-fold in HaCaT cells treated with $10{\mu}g/ml$ of ginsenoside Rb1 for 24 hr, PLA2G4A was increased about twice after 6 hours, and gene expression was increased more than 2-fold after 24 hr. Knockdown of STAT3 with siRNA decreased FAS expression and increased PLA2G4A expression but only FAS was passed from the upstream regulator STAT3. These results indicate that STAT3, which is an upstream regulator, induces apoptosis via FAS during treatment with ginsenoside Rb1.

• Tuberculosis and Respiratory Diseases
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• v.56 no.4
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• pp.381-392
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• 2004

Arsenic trioxide($As_2O_3$) was introduced into the treatment of refractory or relapsed acute promyelocytic Ieukemia. Some investigators have reported that arsenic trioxide had induced apoptosis in a variety of solid human tumor cell lines, including non-small cell lung cancer. Non-steroidal anti-inflammatory drugs(NSAIDs) are powerful chemopreventive agents for gastrointestinal cancers and the growth of established tumors are reduced by inducing apoptosis. It's also reported that NSAIDs enhanced tumor response to chemotherapeutic drugs or radiation. In this study, we aimed to determine whether combination of arsenic trioxide with sulindac augmented its apoptotic potential in NCI-H157 human lung cancer cells. The human lung cancer cell line NCI-H157 was treated with arsenic trioxide and sulindac. Cell viability was measured by the MTT assay. Apoptosis was measured by nuclear staining and flow cytometric analysis. The catalytic activity of the caspase families were measured by the fluorogenic cleavage of biosubstrates. The western blotting were also performed to define the mechanical basis of apoptosis. Combination treatment of arsenic trioxide and sulindac decreased the viability of NCI-H157 human lung cancer cells in a dose-dependent manner. The catalytic activity of caspase-3, 8 and 9 proteases were increased after combination treatment. Consistently PARP was cleaved from 116kDa to 85kDa fragments, and the expression of ICAD was decreased by time-dependent manner. Also combination treatment increased the expression of Fas and Fas/L. Combination therapy of arsenic trioxide with sulindac augments cell death and induces apoptosis via the activation of caspase cascade in NCI-H157 human lung carcinoma cells.

• Journal of Microbiology and Biotechnology
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• v.24 no.10
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• pp.1438-1444
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• 2014

Lactic acid bacteria (LAB) are probiotics that provide numerous beneficial effects on the host body, especially on the intestine. Combining several strains of LAB, we prepared a formulation containing four different LAB and studied its anti-inflammatory activity both in vitro and in vivo. The formulation significantly reduced NO production from RAW 264.7 cells treated with bacterial lipopolysaccharide, indicating that the formulation might include anti-inflammatory activity. The formulation also suppressed inflammatory change induced by trinitrobenzene sulfonic acid (TNBS) in mice, where oral or rectal administration of the formulation protected the colon tissue from the damage by TNBS. Expressions of the IL-6 and FasL genes appeared to be down-regulated by the formulation in TNBS-treated colon tissues, suggesting that the suppression of those genes may be involved in the anti-inflammatory activity of the formulation.

• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1607-1615
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• 2007

We investigated the applicability of the TEM-l ${\beta}$-lactamase fragment complementation (BFC) system to develop a strategy for the screening of protein-protein interactions in bacteria. A BFC system containing a human Fas-associated death domain (hFADD) and human Fas death domain (hFasDD) was generated. The hFADD-hFasDD interaction was verified by cell survivability in ampicillin-containing medium and the colorimetric change of nitrocefin. It was also confirmed by His pull-down assay using cell lysates obtained in selection steps. A coiled-coil helix coiled-coil domain-containing protein 5 (CHCH5) was identified as an interacting protein of human uracil DNA glycosylase (hUNG) from the bacterial BFC cDNA library strategy. The interaction between hUNG and CHCH5 was further confirmed with immunoprecipitation using a mammalian expression system. CHCH5 enhanced the DNA glycosylase activity of hUNG to remove uracil from DNA duplexes containing a U/G mismatch pair. These results suggest that the bacterial BFC cDNA library strategy can be effectively used to identify interacting protein pairs.

• Fisheries and Aquatic Sciences
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• v.6 no.3
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• pp.105-109
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• 2003

Fifty four strains of pathogenic vibrios were isolated from the Gwangan Beach from May to October, 2002. The isolated vibrios were composed of 7 different species: Vibrio parahaemolyticus, V. cholerae non-O1, V. alginolyticus, V. vulnificus, V. hollisae, V. fluvialis, ane V. mimicus. In the detection rate, V. parahaemolyticus was most predominant as $46\%$(25/54). From the isolated strains, only 25 strains have hemolytic activity or 25 strains only proteolytic activity on agar plates. Eleven strains showed both hemolytic and proteolytic activity. No strains showed urease activity. All strains of V parahaemolyticus did not show hemolytic activity, while V. cholerae non-O1 strains showed $\beta$ hemolytic activity. Kanagawa phenomena of pathogenic vibrios did not accord with hemolytic activity of the culture supernatant at the late log phase. Some strains showed high hemolytic activity despite having proteolytic activity, but some weak hemolytic activities despite having no proteolytic activity.

• Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.341-347
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• 2015

We evaluated the biological activities of five water extracts of tissue of the mottled anemone Urticina crassicornis. Most extracts exhibited broad-spectrum antimicrobial activity as determined by ultrasensitive radial diffusion assay (URDA) against gram-positive and -negative bacteria, including a fish pathogen, Aeromonas hydrophila, but no activity against fungi. The activity of the extracts was abolished by tryptic digestion, indicating that protein compounds were responsible for the antimicrobial activity. Furthermore, in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity assay, only the visceral tissue extract showed activity. However, no extract had hemolytic activity against human red blood cells. Consequently, this study suggests the water-soluble extract of mottled anemone to be a promising source of proteinaceous antimicrobial compounds that can be utilized for development of novel antibiotics.

• Neonatal Medicine
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• v.17 no.1
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• pp.44-52
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• 2010

Purpose : The aim of this study was to develop a model for necrotizing enterocolitis (NEC) in the neonatal rat using endotoxin and hypoxia, a plausible insult in a neonatal intensive care and to investigate the role of apoptosis as the underlying mechanism. Methods : Newborn rats were given oral endotoxin and intermittent 8% hypoxia$\pm$caspase inhibitor. The intestinal histology was evaluated using hematoxylin-eosin staining. Apoptosis was analyzed with TUNEL staining and by measuring the caspase 3 activity in the intestinal lysates. IEC-6 cells were assessed for apoptosis and the expression of Bax, Bcl-2, Fas and FasL was measured after treatment with endotoxin and hypoxia. Results : Oral endotoxin (5 mg/kg) and exposure to 8% hypoxia of 60-min duration twice induced human NEC-like lesions in the rat intestine. Intestinal tissue revealed increased apoptosis and caspase-3 activity. After caspase inhibitor treatment, the grades of both apoptosis and NEC were significantly reduced. IEC-6 cells exhibited increased apoptosis and caspase 3 activity after endotoxin and hypoxia treatment and significantly increased Bax/Bcl- 2 ratio compared to control cells. Conclusion : This neonatal rat model of NEC which was induced by oral endotoxin and intermittent hypoxia showed increased apoptosis of intestinal epithelial cells that was mediated by caspase 3 activation. Our model has a advantage in the study of NEC because the use of much more clinically plausible insults may provide a suitable model for the investigation of its pathophysiology and therapeutic trials.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1293-1298
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• 2008

The purpose of this study is to investigate the effect of a mixture of fermented Artemisiae Argi Folium and fermented Sophorae Radix (FAS) on hydrogen peroxide production within mouse macrophage Raw 264.7 with ethanol (EtOH) and nicotine. Artemisiae Argi Folium is known to have the antibacterial, immune-enhancing properties. And Sophorae Radix is also known to have the antibacterial, anti-inflammatory, anti-allergic properties. EtOH and nicotine are the ones of toxicants which could impair immunocytes like macrophage. EtOH and nicotine reduce the intracellular production of hydrogen peroxide ($H_2O_2$) of Raw 264.7. FAS increased the production of hydrogen peroxide reduced by EtOH and nicotine within Raw 264.7. These results indicate that FAS could restore the immuno-activity of macrophage impaired by EtOH and nicotine.