• Title/Summary/Keyword: ${\alpha} -C_2S$

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Exosome-mediated delivery of gga-miR-20a-5p regulates immune response of chicken macrophages by targeting IFNGR2, MAPK1, MAP3K5, and MAP3K14

  • Yeojin Hong;Jubi Heo;Suyeon Kang;Thi Hao Vu;Hyun S. Lillehoj;Yeong Ho Hong
    • Animal Bioscience
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    • v.36 no.6
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    • pp.851-860
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    • 2023
  • Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.

The Role of Lymphocyte Compartment and Cytokine in Coal Workers Pneumoconiosis (진폐증환자에서의 임파구 분획 및 싸이토카인의 역할)

  • Lee, Jung-Yeon;Yoo, Kwang-Ha;Ahn, Hae-Ryon;Kim, Sung-Ryul;Lee, Hae-Woon;Ahn, Cheol-Min;Kim, Hyung-Joong
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.3
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    • pp.241-250
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    • 2002
  • Background : Coal-worker's pneumoconiosis(CWP) is characterized by a chronic inflammatory lung reaction associated with macrophage accumulation in the alveolar spaces. CWP is usually divided into two stage : simple pneumoconiosis(SP) where there are a limited number of fibrotic lesions remain limited, with radiological opacities smaller than 1cm and progressive massive fibrosis(PMF), which is characterized by the development of a perifocal extensive fibrotic response of the lung and severe alterations in pulmonary function. In this study, the lymphocyte compartment and cytokine were evaluated by measuring the serum levels in the control, SP and PMF groups. Materials and Methods : The coal workers selected for this study were employees(patients?) of the Tae-Baek and Dong-Hae hospital. All were men, 45-76 years old and the mean duration of their exposure to coal dust was 23.2 years in the lymphocyte compartment and 24.3 years in the cytokine checked group. According to X-ray examination results, the patients were classified into either one of the SP, PMF categories. The normal controls examined were 26-70 years old men. The serum cytokine levels were estimated by using an end point enzyme immunoassay technique. Results : T lymphocyte, helper and suppressor T cells were highly related to pneumoconiosis in this study. A statistically significant decrease in the number of suppressor T lymphocytes was observed in the simple pneumoconiosis patients and at the same time, there was an increase in the lymphocyte index. Howevere, there was no statistically difference in the serum cytokines levels among the SP, PMF and control groups. Conclusion : T lymphocyte, helper T, and suppressor T cells may be highly related to the development of CWP compared to the control group particularly in the early stage of pneumoconiosis. The changes observed in the immunological system in patients with pneumoconiosis may lie at the bottom of the pathogenesis of fibrosis. Further study is needed to evaluate the lymphocyte compartment as a marker for pneumoconiosis development in the early stage.

Effects of Amylase and Cellulase Supplementation in Sorghum-based Diets for Finishing Pigs

  • Park, J.S.;Kim, I.H.;Hancock, J.D.;Hines, R.H.;Cobb, C.;Cao, H.;Hong, J.W.;Kwon, O.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.1
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    • pp.70-76
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    • 2003
  • Three experiments were conducted to determine the effects of a sorghum-specific enzyme system, derived from an Aspergillus niger and Bacillus subtilis fermentation extract (carbohydrase activity of 1,650 $\alpha$-amylase units and cellulase activity of 30 fibrinolytic units/mL), on growth performance of finishing pigs. In Exp. 1,192 pigs (average initial BW of 46.1 kg) were fed sorghum-based diets without or with 360 mL of enzyme system per ton of sorghum in a 78 d growth assay. For d 0 to 39, gain/feed was improved (p<0.03) with enzyme supplementation, but ADG was not affected (p>0.15). For d 39 to 78 and overall (d 0 to 78), ADG, gain/feed, and digestibilities of DM and N were not affected (p>0.13) by enzyme supplementation. Backfat thickness, fat-free lean index, and scores for stomach keratinization and ulcers also were not affected (p>0.15) by the dietary treatments. In Exp. 2,168 pigs (average initial BW of 58.4 kg) were fed diets without or with 150, 300, or 450 mL/ton of the same enzyme system used in Exp. 1. Adding as much as 450 mL enzyme system / ton of sorghum did not affect (p>0.15) ADG or gain/feed for d 0 to 29 of the growth assay. However, during d 29 to 63, ADG increased by 11% (linear effect, p<0.02) and gain/feed increased by 10% (linear effect, p<0.06) as enzyme concentration was increased from none to 450 mL/ton of sorghum. For the overall period (d 0 to 63), ADG tended to increase (p<0.08) with enzyme supplementation, but gain/feed and digestibilities of DM and N were not affected (p>0.14). Carcass characteristics (dressing percentage, backfat thickness, and fat free lean index) also were not affected (p>0.20) by addition of the enzyme system. In Exp. 3,176 pigs (average initial BW of 46.7 kg) were fed diets without or with 450, 900, or 1,350 mL/ton of the same enzyme system used in Exp. 1 and 2 in a 71 d growth assay. Adding up to 1,350 mL/ton of enzyme had no effects (p>0.15) on ADG, gain/feed, digestibilities of DM and N, and carcass characteristics (dressing percentage, backfat thickness, and fat-free lean index). In conclusion, finishing pigs fed diets with a sorghum-specific enzyme system showed some positive trends for improved growth performance, but those effects were not large and (or) consistent.

VARIETAL DIFFERENCES IN DISTRIBUTION AND PHYSIOLOGICAL CHARACTERISTICS OF RICEROOT (수도근(水稻根)의 분포(分布) 및 생리적(生理的) 특성(特性)에 관(關)한 품종간차이(品種間差異))

  • Park, H.;Park, Y.S.;Kim, Y.W.;Shin, C.S.;Kim, Y.S.
    • Applied Biological Chemistry
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    • v.15 no.1
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    • pp.49-57
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    • 1972
  • Varietal difference in root distribution and other root characteristics were investigated under fold and water culture condition. The results were as follows: 1. IR667 showed funnel type of root distribution in soil profit while Jinheung had barrel type, and each type appearance was more distinguishable with fertilizer application. 2. Root weight per tiller was smaller in IR667 than in Jinheung and IR667 had more root in 0 to 5cm of soil depth but Jinheung had more in 5 to l0cm depth. 3. Horizontal distribution of root was dencer near to stem base without fertilizer than with fertilizer in both IR667 and Jinheung indicating structural construction for intensive nutrient uptake. Between varieties this 'dence to stem base' trend accompaning 'dence to wide spacing side' was greater in IR667 without fertilizer and these were quite true with fertilizer in Jinheung. 4. The decreasing rates of root and ear weight by fertilizer application were greater in IR667 than in Jinheung. This and other characteristics indicated that the root of IR667 is likely to be panicle-number type comparing with Jinheung. 5. The root of IR667 had lower oxidizing power of ${\alpha}-naphthylamine$ than that of Jinheung indicating weaker resistance to reductive soil but cation exchange capacity of water-cultured root was higher in IR667 suggesting stronger nutrient uptake. 6. The content of phosphorus and especially potassium in root were higher with fertilizer but lower without fertilizer in IR667 than in Jinheung indicating that IR667 is more sensitive to root environment. 7. The contents of N, K and CEC were increasing toward root tip while P content was decreasing. The root from surface soil had higher N and K content than that from subsoil. The contents of N,P,K, and CEC of root at harvesting stage were about 1.0%, 0.1%, 0.5% and 15me/100g at dry weight base, respectively.

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Retentive bond strength of fiber-reinforced composite posts cemented with different surface treatments (Fiber reinforced composite post의 표면 처리에 따른 접착 강도)

  • Roh, Hyunsik;Noh, Kwantae;Woo, Yi-Hyung;Pae, Ahran
    • The Journal of Korean Academy of Prosthodontics
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    • v.52 no.2
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    • pp.113-120
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    • 2014
  • This study will evaluate the effectiveness of various pretreatments when fiber-reinforced composite (FRC) post is bonded to endodontically treated tooth with resin cement. Materials and methods: Canal shaping of FRC post (DT Light post, Size 3, Bisco Inc., Schaumburg, IL, USA) was performed on endodontically treated premolars at 1.5 cm from CEJ. Samples were divided into 6 groups of surface treatment after conventional washing and drying to the canal. Total of 24 FRC posts were randomly divided into 6 groups of surface treatment as follows: Group C: control - no surface treatment, Group A: airborne-particle abrasion (Cojet sand, 3M ESPE), Group S: silanization (Bis-silane, Bisco Inc.), Group M: universal primer (Monobond-plus primer, Ivoclar Vivadent Inc.), Group AS: silanization after airborne-particle abrasion, Group AM: universal primer treatment after airborne-particle abrasion. Pretreated fiber posts were cemented with resin-based luting material and photo-polymerized and cut to the thickness of 1 mm. Push-out test using a universal testing machine was performed. Bonding failure strength of post dislodgement was measured and the type of bonding failure was classified. Data were analyzed with Kruskal-Wallis test and multiple comparison groups were performed using Tukey HSD value of rank test (${\alpha}=0.05$). Results: Group AS showed significantly highest bonding strength. Group S, group AM, group A, and group M showed lower bonding strength in order. The control group showed the lowest bonding strength. Conclusion: Surface treatment with silane showed to be the most effective of the surface pretreatment methods for cementation of FRC post. Surface treatment with universal primer showed no significant difference compared with no surface treatment group as for bonding strength.

Development of Monoclonal Antibody to Cytomegalovirus Maior Immediate-early(α) Protein (거대세포바이러스 조기항원 단백질에 대한 단세포군 항체 제조에 관한 연구)

  • Park, Ji Ho;Sohn, Young Mo;Park, Kyu Hyun;Lee, Won Young
    • Pediatric Infection and Vaccine
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    • v.6 no.2
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    • pp.253-260
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    • 1999
  • Purpose : The purpose of this study is to make and use monoclonal Ab which reacts with CMV major immediate early(${\alpha}$) protein(p72). Methods : Normal human fibroblast(Foreskin derived) was cultured in Eagle's minimal essential medium(MEM) containing 10% cowfetus serum and mouse chondroblast was cultured in P3X63 Ag8.653(ATCC. Maryland USA) to maintain $5{\times}10^5/ml$ cell counts. CMV(KJHJ90) from congenital CMV infected infant's urine was multiplied and used for Ab making. CMV Ag was injected 4 times, 1 week interval into the peritoneal space of 6~8 weeks old mice. And then lymphocyles and fibroblasts taken from spleen were obtained and conjugated. After the conjugated cell cultured, we chose the cell that had high Ab titer using indirect immunofluerescent method. Results : Among the 28 monoclonal antibodies obtained LPC12 and LPC23 reacted highly with nucleus of AD169 infected cell. Purified AD169 after SDS-PAGE, molecular weight of Ag, which reacted with purified monoclonal Ab, was obtained using Western blotting. Monoclonal Ab of LPC12 and LPC23 clone reacted most highly with 72 kd Ag. Conclusion : LPC12 and LPC23 clonal Ab with AD 169(P63-27) is useful on early diagnosis of CMV infection.

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Construction and Analysis of a DNA Microarray for the Screening of Biosynthetic Genes of Secondary-Metabolites formation in Streptomyces (방선균 유래 이차대사 생합성 유전자 분석용 DNA Microarray 제작 및 해석)

  • Nam Soo Jung;Kang Dae-Kyung;Rhee Ki Hyeong;Kim Jong-Hee;Kang Sang Sun;Chang Yong Keun;Hong Soon-Kwang
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.105-111
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    • 2005
  • Streptomyces produces many kinds of secondary-metabolites including antibiotics. Screening of a new compound and elucidation of a biosynthetic pathway for the secondary metabolites are very important fields of biology, however, there is a main problem that most of the identified compounds are already researched compounds. To solve these problems, a microarray system that is based on the data related to the biosynthetic genes for secondary-metabolites was designed. For the main contents of DNA microarray, the important genes for the bio-synthesis of aminoglycosides, polyenes group, enediyne group, alpha-glucosidase inhibitors, glycopeptide group, and orthosomycin group were chosen. A DNA microarray with 69 genes that were involved in the bio-synthesis for the antibiotics mentioned above was prepared. The usability of the DNA microarray was confirmed with the chromosomal DNA and total RNA extracted from S. coelicolor whose genomic sequence had already been reported.

Damage Study on the Mechanical Fastening in Laminated Composites (복합적층판(復合積層板)의 기계적(機械的) 체결부(締結部)에 관한 파손연구(破損硏究))

  • Kwan-Hyung,Song
    • Bulletin of the Society of Naval Architects of Korea
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    • v.27 no.4
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    • pp.58-66
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    • 1990
  • A series of test was performed measuring the failure strength and failure mode of Gr/Pi, $[0^{\circ}/45^{\circ}/90^{\circ}/-45^{\circ}]_s$ laminate containing a single pin loaded hole. The finite element method is applied to calculate the stress distribution in the laminates, then the failure load and the failure mode were predicted by means of the characteristic length. 12 different geometric variations were developed to analyze the effects of the ratio of specimen width to hole diameter (W/d) and ratio of edge distance to hole diameter (L/d). X-Ray of NDE methods were utilized in finding out the initial defects, damage and the fracture mechanism, and SEM(Scanning Electron Microscopes) was used the evaluation of the fracture mechanism and crack propagation around hole under tension pin loading. $[0^{\circ}/45^{\circ}/90^{\circ}/-45^{\circ}]_s$ laminate are found to be most sensitive to W/d but not so influenced by L/d. The failure mode and tensile strength predicted by the model show agreement with experiment data for pin loading bolted jointed test except range of $L/d{\leqq}3$.

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Combined Effect of Ganciclovir and Vidarabine on the Replication, DNA Synthesis, and Gene Expression of Acyclovir-resistant Herpes Simplex Virus (Acyclovir저항성 Herpes Simplex Virus의 복제, DNA합성 및 형질 발현에 미치는 Ganciclovir 및 Vidarabine의 병용효과에 관한 연구)

  • Yang, Young-Tai;Cheong, Dong-Kyun;Mori, Masakazu
    • The Korean Journal of Pharmacology
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    • v.25 no.1
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    • pp.115-134
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    • 1989
  • Combined effects of ganciclovir (GCV) and vidarabine (ara-A) on the replication, DNA synthesis, and gene expression of wild type-1 herpes simplex virus (HSV-1) and three acyclovir (ACV)-resistant HSV-1 mutants were studied. These mutants include a virus expressing no thymidine kinase $(ACV^r)$, a virus expressing thymidine kinase with altered substrate specificity $(IUdR^r)$, and a mutant expressing altered DNA polymerase $(PAA^r5)$. GCV, an agent activated by herpesvirus specific thymidine kinase, showed potent antiviral activity against the wild type HSV-1(KOS) and DNA polymerase mutant $(PAA^r5)$. The ACV-resistant mutants with thymidine kinase gene $(ACV^r\;and\;IUdR^r)$ were resistant to GCV. All tested wild type HSV-1 or ACV-resistant HSV-1 mutants did not display resistance to vidarabine (are-A). Combined GCV and ara-A showed potentiating synergistic antiviral activity against wild type KOS and $PAA^r5$, and showed subadditive combnined ativiral activity against thymidine kinase mutants. Combined GCV and ara-A more significantly inhibited the viral DNA synthesis in wild type KOS and $PAA^r5-infected$ cells to a greater extent than either agent alone, but the synergism was not determined in $ACV^r$ or $IUdR^r-infected$ cells. These data clearly indicate that combined GCV and ara-A therapy might be useful for the treatment of infections caused by wild type HSV-1 or ACV-resistant HSV-1 with DNA polymerase mutation. ACV-resistant viruses with the mutation in thymidine kinase gene are also, resistant to GCV, but susecptible to ara-A, indicating that ara-A would the drug of choice for the treatment of ACV-resistant HSV-1 which does not express thymidine kinase or expresses thymidine kinase with altered substrate specificity. While the synthesis of viral ${\alpha}-proteins$ of wild type HSV-1 was not affected by ACV, GCV, ara-A, or combined GCV and ara-A, the synthesis of ${\beta}-proteins$ was slightly but significantly increased at the later stage of viral infection by the antiviral agents. The synthesis of ${\gamma}-proteins$ of wild type HSV- 1 was significantly inhibited by ACV, GCV, ara-A, and combined GCV and ara-A. Combined GCV $(5-{\mu}M)$ and ara-A $(100-{\mu}M)$ also significantly altered the expression of viral ${\beta}-and$ ${\gamma}-proteins$, of which efffct was similar to that of GCV $(10-{\mu}M)$ alone. Although ACV at the concentration of $10-{\mu}M$ did not alter the expression of ${\alpha}-$, ${\beta}-$, and ${\gamma}-proteins$ of ACV-resistant $PAA^r5$, GCV and ara-A significantly alter the epression of ${\beta}-and$ ${\gamma}-proteins$, not ${\alpha}-protein$, as same manner as they altered the expression of those proteins in cells inffcted with wild type HSV-1. Combined GCV $(5-{\mu}M)$ and ara-A $(100-{\mu}M)$ altered the expression ${\beta}-and$ ${\gamma}-proteins$ in $PAA^r5$ infected cells, and the effect of combined regimen was comparable of that of GCV $(10-{\mu}M)$. These data indicate that the alteration in the expression of ${\beta}-and$ ${\gamma}-proteins$ in wild type HSV-1 or $PAA^r5$ infected cells could be more significantly affected by combined GCV and are-A than individual GCV or ara-A. In view of the fact that (a) viral ${\alpha}-$, ${\beta}-$, and ${\gamma}-proteins$ are synthesized in a cascade manner; (b) ${\beta}-proteins$ are essential for the synthesis of viral DNA; (c) the synthesis of ${\beta}-proteins$ are inhibited by ${\gamma}-proteins$; and (d) most ${\gamma}-proteins$ are made from the newly synthesized progeny virus, it is suggested that GCV and ara-A, alone or in combination, primarily inhibit the synthesis of viral DNA, and by doing so might exhibit their antiherpetic activity. The alteration in viral protein synthesis in the presence of tested antiviral agents could result from the alteration in viral DNA synthesis. From the present study, it can be concluded that (a) combined GCV and ara-A therapy would be beneficial for the control of inffctions caused by wild type HSV-1 or ACV-resistant DNA polymerase mutants; (b) the combined synergistic activity of GCV and ara-A is due to further decrease in the viral DNA by the combined regimen; (c) ara-A is the drug of choice for the infection caused by ACV-resistant HSV-1 with thymidine kinase mutation; and (d) the alteration in viral protein synthesis by GCV and ars-A, alone or in combination, is mostly due to the decreased synthesis of viral DAN.

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Effects of Green Tea Seed Oil on the Cholesterol, TBARS and Inflammatory Responses in C57BL/6 Mice Fed High Cholesterol Diet (녹차씨기름이 고콜레스테롤식이를 섭취시킨 C57BL/6 Mice의 콜레스테롤, TBARS 및 염증반응에 미치는 영향)

  • Kim, Jong-Kyung;Kim, So-Hee;Noh, Kyung-Hee;Jang, Ji-Hyun;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.3
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    • pp.284-290
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    • 2007
  • Effects of green tea seed oil intake on the serum cholesterol, the thiobarbituric reactive substances (TBARS) formation of liver and inflammatory markers of plasma and macrophage in cholesterol fed mice were investigated, comparing to the intakes of corn oil and olive oil. C57BL/6 mice were divided into three groups and fed the experimental diets: supplemented corn oil, green tea seed oil and olive oil to cholesterol diet, respectively, at the level of 10% for 9 weeks. The increased levels of serum cholesterol of green tea seed oil group were significantly (p<0.05) lower than that of corn oil group until 6 weeks. The TBARS formation in liver of green tea seed oil and olive oil groups were significantly (p<0.05) reduced than those of corn oil group. The contents of inflammatory markers ($LTB4,\;TNF-{\alpha},\;PGE2,\;NO$) in plasma and macrophage did not show significant differences among the experimental groups. These results showed that green tea seed oil could exert the activity of decreasing serum cholesterol and the antioxidative activity in cholesterol fed mice liver. Therefore, we suggest that green tea seed oil might be developed as a high quality edible oil.