• Title/Summary/Keyword: HEP method

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Effect of Growth Inhibition and Quinone Reductase Activity Stimulation of Makgeoly Fractions in Various Cancer Cells (막걸리 분획물에 의한 암세포 성장 억제 및 Quinone Reductase 활성 증가 효과)

  • Shin, Mi-Ok;Kang, Dae-Yeon;Kim, Mi-Hyang;Bae, Song-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.3
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    • pp.288-293
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    • 2008
  • In this study, we investigated the anticancer activity of Makgeoly (MG). MG was fractionated into four fractions by using solvent partition method, affording hexane (MGH), methanol (MGM), butanol (MGB) and aquous (MGA) soluble fractions. We determined the cytotoxicity of these four fractions in four kinds of cancer cell lines, such as HepG2, MCF-7, B16-F10 and HT29 by MTT assay. Among the various fractions, the MGM showed the strongest cytotoxic effects on all cancer cell lines. The morphological changes such as membrane shrinking and blebbing of cells were also observed by MGM treatment in HepG2 cell. In addition, we observed quinone reductase (QR) activity stimulating effects in all fraction layers of MG on HepG2 cells. QR activity increased approximately 2.6 and 2.1 times in MGM and MGH treated HepG2 cell at $100{\mu}g/mL$, respectively, compared to that in control value. Although further studies are needed, the present work could suggest that the fin of MG has a potential to be used as a chemopreventive agent against cancer.

Screening of Differentially Expressed Genes by Desferrioxamine or Ferric Ammonium Citrate Treatment in HepG2 Cells

  • Park, Jong-Hwan;Lee, Hyun-Young;Roh, Soon-Chang;Kim, Hae-Yeong;Yang, Young-Mok
    • BMB Reports
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    • v.33 no.5
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    • pp.396-401
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    • 2000
  • A differential display method is used to identify novel genes whose expression is affected by treatment with ferric ammonium citrate (FAC) or desferrioxamine (DFO), an iron chelating agent in the human hepatoblastoma cell line (HepG2). These chemicals are known to deplete or increase the intracellular concentration of iron, respectively. Initially, we isolated seventeen genes whose expressions are down- or up regulated by the treatment of the chemicals, as well as their four differentially expressed genes that are designated as clone-1, -2, -3, and -4. These are further characterized by cDNA sequencing and Northern blot analysis. Through the cDNA sequencing, as well as comparing them to genes published using the NCBI BLAST program, we identified the sequence of the clone-1 that is up-regulated by the treatment of DFO. It is identical to the human insulin-like growth factor binding protein-1 (IGFBP-1). This suggests that the IGFBP-1 gene in the HepG2 cell is up-regulated by an iron depletion condition. Also, the expression of the clone-3 and -4 is up-regulated by FAC treatment and their eDNA sequences are identical to the human ferritin-fight chain and human NADH-dehydrogenase, respectively. However, the sequence of the clone-2 has no significant homology to any other known gene. Therefore, we suggest that changes of the cellular iron level in the HepG2 cell affects the transcription of cellular genes. This includes human IGFBP-1, ferritin-fight chain, and NADH-dehydrogenase. Regulation of these gene expressions may have an important role in cellular functions that are related to cellular iron metabolism.

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The Effect of Hepatitis Treatment-Tang No.1 on Ethanol-Induced Cytotoxicity of Hep G2 Cells (Hep G2 세포에서 간염제1탕의 에탄올에 의한 세포독성 억제효과)

  • Park, Young-Kweon;Kim, Gang-San;Kang, Byung-Ki;Ra, Ki-Ung
    • The Journal of Internal Korean Medicine
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    • v.22 no.1
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    • pp.79-85
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    • 2001
  • Object : Hepatitis Treatment-tang No.1 has been used for the treatment of Liver disease and Jaundice. Long-term EtOH exposure leads to immunoregulatory and detoxification impairment. This study aimed to determine the relationship between TNF-${\alpha}$ production and expression, and EtOH-induced cytotoxicity on Hep G2 cells. Method : Cells were incubated with EtOH in the presence or absence of HT. The cells were tested after 24 hours and, again, after 48 hours. Cytoviability and TNF-${\alpha}$ release were analyzed by MTT assay and enzyme linked immunosorbent assay (ELISA), respectively. After 24 hours of EtOH exposure, the cytoviability decreased, and the release of TNF-${\alpha}$ was increased. Increased amounts of TNF-${\alpha}$ contribute to EtOH-induced cytotoxicity. The Anti-TNF-${\alpha}$ antibody almost abolished it. Interestingly, EtOH-induced cytotoxicity and TNF-${\alpha}$ production were inhibited by HT. Moreover, when HT was used in combination with the anti-TNF-${\alpha}$ antibody, there was a marked inhibition of EtOH-induced cytotoxicity. Results : These results suggest that HT may prevent the cytotoxicity through partial inhibition of the TNF-${\alpha}$ secretion.

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Differential Gene Expression Induced by Naphthalene in Two Human Cell Line, HepG2 and HL-60

  • Kim, Youn-Jung;Song, Mee;Song, Mi-Kyung;Youk, Da-Young;Choi, Han-Saem;Sarma, Sailendra Nath;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.5 no.2
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    • pp.99-107
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    • 2009
  • Naphthalene is bicyclic aromatic compound that is widely used in various domestic and commercial applications including lavatory scent disks, soil fumigants and moth balls. Exposure to naphthalene results in the development of bronchiolar damage, cataracts and hemolytic anemia in humans and laboratory animals. However, little information is available regarding the mechanism of naphthalene toxicity. We investigated gene expression profiles and potential signature genes in human hepatocellular carcinoma HepG2 cells and human promyelocytic leukemia HL-60 cells after 3 h and 48 h incubation with the IC$_{20}$ and IC$_{50}$ of naphthalene by using 44 k agilent whole human genome oligomicroarray and operon human whole 35 k oligomicroarray, respectively. We identified 616 up-regulated genes and 2,088 down-regulated genes changed by more than 2-fold by naphthalene in HepG2 cells. And in HL-60, we identified 138 up-regulated genes and 182 down-regulated genes changed by more than 2-fold. This study identified several interesting targets and functions in relation to naphthalene-induced toxicity through a gene ontology analysis method. Apoptosis and cell cycle related genes are more commonly expressed than other functional genes in both cell lines. In summary, the use of in vitro models with global expression profiling emerges as a relevant approach toward the identification of biomarkers associated with toxicity after exposure to a variety of environmental toxicants.

A Suitability Selection for Marine Afforestation Using Habitat Evaluation Procedure (서식지 평가 방법을 이용한 바다숲 조성 해역의 적지 선정)

  • Oh, Tae-Geon;Kim, Yi-Cheong;Yang, Yong-Su;Kim, Chang-Gil;Lee, Moon-Ock
    • Journal of Advanced Marine Engineering and Technology
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    • v.34 no.6
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    • pp.894-905
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    • 2010
  • A habitat evaluation method was used to estimate the optimum suitability of the study area for the target algae. Habitat evaluation was carried out using an habitat evaluation procedure (HEP) so that the optimum suitability was quantitatively estimated for carrying out marine afforestation in the study area. According to the results of the suitability analysis, the variation of light and wave conditions according to depth showed the factors with the largest impact to involve the spatial distribution of suitable locations within the area. The total suitable area selected was calculated to be 18ha. The quality of the target algae (Ecklonia cava Kjellman) habitat was analyzed using an habitat suitability index (HSI) model of the HEP, which showed 0.55-0.907 (the maximum value being 1.0). This indicated that artificial reefs for afforestation should be installed to zonation type because the suitable area selected (The HSI value was 0.55~0.907) was distributed within the same depth line.

Effect of Injin Fraction on Hepatic Fibrosis induced by $TGF-{\beta}1$ (인진이 $TGF-{\beta}1$ 유도성 간섬유화에 미치는 영향)

  • 신성만;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.22 no.3
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    • pp.141-155
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    • 2001
  • Objective : The aim of this study is to investigate the effect of Injin fractions on hepatic fibrosis induced by $TGF-{\beta}1$. Method : $TGF-{\beta}1$ mRNA, protein, $TGF-{\beta}1$ receptor, Smad family and PAI-I mRNA were studied in HepG2 cell, and the proliferation, connective tissue growth factor, fibronectin and collagen type I mRNA in T3891 fibroblast by quantitative RT-PCR, ELISA and thymidine incorporation assay. Results : On $TGF-{\beta}1$ mRNA and protein synthesis in HepG2, $H_2O$, butanol and hexane fractions of Injin showed inhibitory effect in a dose-dependent way. In the study on $TGF-{\beta}1$ receptor, Smad family and PAI-1 mRNA in HepG2, $H_2O$, butanol and hexane fraction of Injin showed inhibitory effect on the expression of PAI-1 in a dose-dependent way. On the proliferation of T3891 fibroblast induced by $TGF-{\beta}1$, $H_2O$, ethylacetate and butanol fractions of Injin showed inhibitory effect. In the study on the factors affected by $TGF-{\beta}1$, $H_2O$, ethylacetate and butanol fractions of Injin showed inhibitory effect on CTGF, and $H_2O$, butanol, chloroform and hexane fractions showed inhibitory effect on the expression of collagen type I, whereas no fraction showed inhibitory effect on the expression of fibronectin Conclusion : These results show that each fraction of Injin acts as a fibrosis inhibitory factor by itself or in combination, ultimately inhibiting liver cirrhosis.

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Identification of Histone Deacetylase 1 Protein Complexes in Liver Cancer Cells

  • Farooq, Muhammad;Hozzein, Wael N.;Elsayed, Elsayed A.;Taha, Nael A.;Wadaan, Mohammad A.M.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.915-921
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    • 2013
  • Background: Hepatocellular carcinoma is one of the leading causes of mortalities worldwide. The search for new therapeutic targets is of utmost importance for improved treatment. Altered expression of HDAC1 in hepatocellular carcinoma (HCC) and its requirement for liver formation in zebrafish, suggest that it may regulate key events in liver carcinogenesis and organogenesis. However, molecular mechanisms of HDAC1 action in liver carcinogenesis are largely unknown. The present study was conducted to identify HDAC1 interacting proteins in HepG2 cells using modified SH-double-affinity purification coupled with liquid mass spectrophotemetery. Materials and Methods: HepG2 cells were transfected with a construct containing HDAC1 with a C-terminal strepIII-HA tag as bait. Bait proteins were confirmed to be expressed in HepG2 cells by western blotting and purified by double affinity columns and protein complexes for analysis on a Thermo LTQ Orbitrap XL using a C18 nano flow ESI liquid chromatography system. Results: There were 27 proteins which showed novel interactions with HDAC1 identified only in this study, while 14 were among the established interactors. Various subunits of T complex proteins (TCP1) and prefoldin proteins (PFDN) were identified as interacting partners that showed high affinity with HDAC1 in HepG2 cells. Conclusions: The double affinity purification method adopted in this study was very successful in terms of specificity and reproducibility. The novel HDAC1 complex identified in this study could be better therapeutic target for treatment of hepatocellular carcinoma.

Implementation of Multilevel Boost DC-Link Cascade based Reversing Voltage Inverter for Low THD Operation

  • Rao, S. Nagaraja;Kumar, D.V. Ashok;Babu, Ch. Sai
    • Journal of Electrical Engineering and Technology
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    • v.13 no.4
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    • pp.1528-1538
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    • 2018
  • In this paper, configuration of $1-{\phi}$ seven-level boost DC-link cascade based reversing voltage multilevel inverter (BDCLCRV MLI) is proposed for uninterrupted power supply (UPS) applications. It consists of three level boost converter, level generation unit and full bridge circuit for polarity generation. When compared with conventional boost cascaded H-bridge MLI configurations, the proposed system results in reduction of DC sources, reduced power switches and gate drive requirements. Inverter switching is accomplished by providing appropriate switching angles that is generated by any optimization switching angle techniques. Here, round modulation control (RMC) method is taken as the optimization method and switching angles are derived and the same is compared with various switching angles methods i.e., equal-phase (EP) method, and half-equal-phase (HEP) method which results in improved quality of obtained AC power with lowest total harmonic distortion (THD). Reduction in DC sources and switch count makes the system more cost effective. A simulation and prototype model of $1-{\phi}$ seven-level BDCLCRV MLI system is developed and its performance is analyzed for various operating conditions.

Anticancer Effect of Extracts from the Marine and Salted Fish Products. (수산물 및 수산 발효식품의 암세포 억제효과)

  • 임현수;김수현;유은정;강동수;최명락;송상호
    • Journal of Life Science
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    • v.11 no.1
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    • pp.48-53
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    • 2001
  • This study was performed to observe the cytotoxic effect of the various salted fish extracts against cancer cell line, human hepatocellular carcinoma (HepG2) using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) method. Urechis unicinctus was the strongest cytotoxic effects among any other traditional salted fish products. The growth inhibition ratio of Urechis unicinctus hot-water extracts was 94.5% at the concentration of 1000$\mu\textrm{g}$/$m\ell$. On the other hand, in case of salted fish methanol extracts, salt-fermented shad gizzard was showed the strongest cytotoxic effects. The growth inhibition ratio of salt-fermented shad gizzard methanol extracts was investigated 90% at the concentration of 1000$\mu\textrm{g}$/.$m\ell$.

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Effect of Endothelial Cell Growth Factor and Cyclic AMP Increasers on the Proliferation of Human Omental Microvascular Endothelial Cells (사람의 대망미세혈관내피세포 증식에 대한 내피세포성장인자 및 CYCLIC AMP 증가물질의 효과)

  • Kim, Won-Gon;Kim, Jong-Man;Yu, Se-Yeong
    • Journal of Biomedical Engineering Research
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    • v.16 no.4
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    • pp.463-470
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    • 1995
  • Complete prelining of artificial vascular grafts with autologous endothelial cells may be one of the ideal solutions to obtain a nonthrombogenlc blood-contacting surface. To establish an intact endothelial cell monolayer on a prosthetic surface at the time of implantation,a sufficient number of endothelial cells and adequate propagation condition In cell culture are prerequisites. In this experimental study, endothelial cells from microvessels of adult human oriental adipose tissue were enzymatically harvested, and optimal culture conditions for proliferation of the endothelial cells in cell culture were examined. Human oriental adipose tissue was digested with collagenase and endothelial cells were separated from other stromal elements by mesh filtration method. Cultured cells were identified as endothelial cells by immunofluorescent staining for factor VIII-related antigen. Proliferation in usual 20% fetal bovine serum (FBS) medium or medium containing endothelial cell growth factor (ECGF)(5 ng/ml) and heparin (HEP)(1,000 units/ml) were compared,and the effects of adding compounds that increase intracellular cyclic adenosine monophosphate levels, that is,cholera toxin (CT)(1 $\mu\textrm{g}$/ml) and isobutylmethylxanthine (IBMX)(0.2 ml),were also analyzed. In total,following eight media groups were examined. 1) FBS medium + ECGF + HEP, 2) FBS medium + ECGF + HEP+CT, 3) FBS medium+ECGF+HEP+lBMX, 4) FBS medium+ECGF+HEP+CT+ IBMX, 5) FBSmedium, 6) FBS medium +CT, 7) FBS medium + IBMX, 8) FBS medium + CT + IBMX. It was shown that the medium containing ECGF + HEP with or without cholera toxin was most efficient in Stimulating cell proliferation. IBMX was considered to have antagonistic effect to ECGF. Among experimental groups without ECGF and HEP, the addition of cholera toxin and IBMX was shown to significantly potentiate cell proliferation. This results could provide a practical method for use of cultured human endothelial cells for endothelial cell seeding of cardiovascular prosthetic device, particularly in small-diameter vascular grafts.

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