• Title/Summary/Keyword: mediator effect

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Vaccinium bracteatum Thunb. Exerts Anti-Inflammatory Activity by Inhibiting NF-κB Activation in BV-2 Microglial Cells

  • Kwon, Seung-Hwan;Ma, Shi-Xun;Ko, Yong-Hyun;Seo, Jee-Yeon;Lee, Bo-Ram;Lee, Taek Hwan;Kim, Sun Yeou;Lee, Seok-Yong;Jang, Choon-Gon
    • Biomolecules & Therapeutics
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    • v.24 no.5
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    • pp.543-551
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    • 2016
  • This study was designed to evaluate the pharmacological effects of Vaccinium bracteatum Thunb. methanol extract (VBME) on microglial activation and to identify the underlying mechanisms of action of these effects. The anti-inflammatory properties of VBME were studied using lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. We measured the production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2, prostaglandin $E_2$ ($PGE_2$), tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-1 beta (IL-$1{\beta}$), and interleukin-6 (IL-6) as inflammatory parameters. We also examined the effect of VBME on intracellular reactive oxygen species (ROS) production and the activity of nuclear factor-kappa B p65 (NF-${\kappa}B$ p65). VBME significantly inhibited LPS-induced production of NO and $PGE_2$ and LPS-mediated upregulation of iNOS and COX-2 expression in a dose-dependent manner; importantly, VBME was not cytotoxic. VBME also significantly reduced the generation of the pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. In addition, VBME significantly dampened intracellular ROS production and suppressed NF-${\kappa}B$ p65 translocation by blocking $I{\kappa}B-{\alpha}$ phosphorylation and degradation in LPS-stimulated BV2 cells. Our findings indicate that VBME inhibits the production of inflammatory mediators in BV-2 microglial cells by suppressing NF-${\kappa}B$ signaling. Thus, VBME may be useful in the treatment of neurodegenerative diseases due to its ability to inhibit inflammatory mediator production in activated BV-2 microglial cells.

UBE2Q1 in a Human Breast Carcinoma Cell Line: Overexpression and Interaction with p53

  • Shafiee, Sayed Mohammad;Rasti, Mozhgan;Seghatoleslam, Atefeh;Azimi, Tayebeh;Owji, Ali Akbar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3723-3727
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    • 2015
  • The p53 tumor suppressor protein is a principal mediator of growth arrest, senescence, and apoptosis in response to a broad array of cellular damage. p53 is a substrate for the ubiquitin-proteasome system, however, the ubiquitin-conjugating enzymes (E2s) involved in p53 ubiquitination have not been well studied. UBE2Q1 is a novel E2 ubiquitin conjugating enzyme gene. Here, we investigated the effect of UBE2Q1 overexpression on the level of p53 in the MDA-MB-468 breast cancer cell line as well as the interaction between UBE2Q1 and p53. By using a lipofection method, the p53 mutated breast cancer cell line, MDA-MB-468, was transfected with the vector pCMV6-AN-GFP, containing UBE2Q1 ORF. Western blot analysis was employed to verify the overexpression of UBE2Q1 in MDA-MB-468 cells and to evaluate the expression level of p53 before and after cell transfection. Immunoprecipitation and GST pull-down protocols were used to investigate the binding of UBE2Q1 to p53. We established MDA-MB-468 cells that transiently expressed a GFP fusion proteins containing UBE2Q1 (GFP-UBE2Q1). Western blot analysis revealed that levels of p53 were markedly lower in UBE2Q1 transfected MDA-MB-468 cells as compared with control MDA-MB-468 cells. Both in vivo and in vitro data showed that UBE2Q1 co-precipitated with p53 protein. Our data for the first time showed that overexpression of UBE2Q1can lead to the repression of p53 in MDA-MB-468 cells. This repression of p53 may be due to its UBE2Q1 mediated ubiquitination and subsequent proteasome degradation, a process that may involve direct interaction of UBE2Q1with p53.

Effect of Heparin-binding Epidermal Growth Factor (HB-EGF) on Integrin $\alpha_{\nu}-\betaFe_3$ Expression in Preimplantation Mouse Embryos

  • Lim, Jung-Jin;Shin, Hyun-Sang;Lee, Ji-Won;Kang, Sue-Man;Lee, Sung-Eun;Kang, Han-Seung;Kim, Moon-Kyoo
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.102-102
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    • 2002
  • Heparin-bindin epidermal growth factor (HB-EGF) is one of the EGF family to be expressed at the time of implantation in the mouse uterus. Although HB-EGF has been shown to stimulate the development of embryo and uterus in the mouse, its correlation between cell adhesion molecules remains undefined. Integrin $\alpha$$_{ν}$$\beta$$_3$, one of the cell adhesion molecules, is an important mediator of cell-substratum and cell-cell adhesion in implantation. In the present studies, we investigated the effects of HB-EGF on the embryonic development, initiation of implantation and expression of integrin $\alpha$$_{ν}$$\beta$$_3$ in in vitro culture, blocking of HB-EGF, RT-PCR and immunofluores cence analysis. The results showed that HB-EGF significantly improved the developmental rate of hatched embryos (24.1%, p<0.01) and outgrowth embryos (42.5%, p<0.01). On the other hand, this growth factor showed no offset before the hatching embryonic stage. Analysis of RT-PCR showed that HB-EGF upregulated the expression level of integrina $\alpha$$_{ν}$$\beta$$_3$ subunit genes on the preimplantation embryo and outgrowth of blastocyst (120hr and 144hr after hCG injection). Immunofluorescence analysis showed that the integrin $\alpha$$_{ν}$$\beta$$_3$ subunits localized at the pericellular borders and cell-cell contact areas. Increase in fluorescence intensity was observed in the HB-EGF treated embryos. Intrauterine injection of an anti-HB-EGF antiserum at day 3 significantly decreased the number of implantation sites (14.4, p<0.01) and significantly increased the number of recovered embryos(6.4, p<0.05) at day 5. From these results, it imply that HB-EGF improve the embryo development and accelerated the expression of integrin $\alpha$$_{ν}$$\beta$$_3$ in the preimplantation mouse embryos.

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The Effect of Platelet Rich Plasma Combined with Bovine Bone on the Treatment of Grade II Furcation Defects in Beagle Dogs (성견 2급 치근이개부 병변 치료시 이종골 이식 및 혈소판 농축 혈장의 골재생에 관한 효과)

  • Lim, Sung-Bin;Lee, Kwang-Soo;Park, Young-Chae;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.257-277
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    • 2000
  • New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration are basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, and biological mediators. Platelet Rich Plasma has been reported as a biological mediator which regulates activities of wound healing progress including cell proliferation, migration, and metabolism. The purpose of this study is to evaluate the effects of using the Platelet Rich Plasma as a regeneration promoting agent for furcation involvement defect. Five adult beagle dogs were used in this experiment. The dogs were anesthetized with Ketamin HCl(0.1 ml/kg, IV)and Xylazine hydrochloride($Rompun^{(R)}$, Bayer, 0.1 ml/kg, IM) and conventional periodontal prophylaxis were performed with ultrasonic scaler and hand instruments. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree II furcation defect was made on mandibular third(P3), forth(P4) and fifth(P5) premolar, and stopping was inserted. After 4 weeks, stopping was removed, and bone graft was performed. Ca-P was grafted in P3(experimental group I), Combination of Ca-P and plasma rich platelet were grafted in P4(experimental group II), and P5 was remained at control group.Systemic antibiotics(gentamicin sulfate)and anlgesics(phenyl butazone) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operate sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 4, 8 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with Gomori's trichrome staining. At 4 weeks after surgery, there were rapid osteogenesis phenomenon on the defected area of the Platelet Rich Plasma plus Ca-P BBP group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. In conclusion, Platelet Rich Plasma can promote rapid osteogenesis during healing of periodontalregeneration.

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A Study of Entrepreneurship Education on Entrepreneurial Orientation of Korean and Chinese University Students: Focused on Entrepreneurial Self-Efficacy as Mediator (기업가정신교육이 한·중대학생들의 기업가적 지향성에 미치는 영향: 자기효능감의 매개효과를 중심으로)

  • Yang, Jun Hwan
    • Asia-Pacific Journal of Business Venturing and Entrepreneurship
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    • v.15 no.3
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    • pp.233-242
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    • 2020
  • This empirical study was designed to identify the key factors that affect entrepreneurial orientation of university students. An exploratory model was suggested to verify the relationship among the research variables including entrepreneurship education, entrepreneurial self-efficacy, and entrepreneurial orientation(Innovativeness, Proactiveness, Risk-Taking Propensity) in order to attain the purpose of the study. This paper also aimed at investigating the possible difference between Korea and China in entrepreneurial process within this research framework. Total 440 university students participated in this study from both countries; Korea(n=224) and China(n=216). The hierarchical multiple regression was employed for estimating technique by using IBM SPSS statistics Version 23. The findings showed that entrepreneurship education works as a key predictor of entrepreneurial orien- tation and entrepreneurial self-efficacy as well. Also, entrepreneurial self-efficacy has a positive and significant mediating effect and goes on to further impact entrepreneurial orientation. So, entrepreneurship education should be enhanced extensively in order to promote entrepreneurship of young people. The theoretical and practical implication were presented.

A Relationship Study of Adolescents' Various Stress, School-life Adjustment, and Happiness: mediating effects (남녀청소년의 다양한 스트레스(부모, 학업, 친구, 외모, 경제), 학교생활적응과 행복감의 관계성 연구: 매개관계를 중심으로)

  • Kim, Sunah
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.9
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    • pp.161-169
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    • 2020
  • This study investigated the effects and relationships of various stress, school life adjustment and happiness factors among middle school and high school adolescents. The structural model group effects between boy and girl groups were also investigated. Data was employed from the 'Korean Children and Youth Well-Being Index Survey, 2018' while subjects of the analysis included 5,144 students. Based on the education statistics data, the sample was collected by probability proportional to size. Results utilizing structural equation modeling and multigroup analysis showed that first, school stress, friend stress, and look stress had significant effects on adolescent happiness. School life adjustment had positive significant effects on adolescents' happiness. Parent stress, money stress and school stress had significant effects on school life adjustment. Second, the mediating effect of school life adjustment between stress and happiness was significant with parent, school stress, and financial stress. Third, results of the multi-group analysis by level and gender showed that the structure was similar between boys and girls, with the only differences among mediating effects. Boys had significant mediating effects by school stress, and financial stress, while girls had significant mediating effects by school stress, financial stress, and parent stress. Finally, based on the results of the study, practice and policy implications were suggested to support the promotion of adolescents' happiness.

Effects of Mahaenggamseok-tang-gagambang on Immune Cells and Cytokines in OVA-Induced Asthmatic Mice (마행감석양가감방(麻杏甘石揚加減方)이 천식모델생쥐의 면역세포 및 사이토카인에 미치는 영향)

  • Park, Gil-Byeong;Park, Yang-Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.3
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    • pp.590-598
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    • 2009
  • The purpose of this research is to evaluate the effect of Mahaenggamseok-tang-gagambang (MGTG) on airway hyper- responsiveness (AHR), immune cells, cytokines and lung tissue in OVA-induced asthmatic mice. C578L/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (3times a week) for asthma sensitization and challenge. Two experimental groups were treated with different concentrations of MGTG (400 mg/kg and 200 mg/kg) extract and cyclosporin A (10 mg/kg) for the later 8 weeks. Enhanced pause (Penh) levels were measured by whole body plethysmography. Immune cells were analyzed by flow cytometer in peripheral blood monocyte cell (PBMC) and lung cells. The IL-1b, IL-12, IFN-${\gamma}$, OVA-lgE, IL-4, IL-5, TNF-${\alpha}$ were analyzed by ELISA kit in serum and splenocyte+a-cCDS/a-CD28. Enhanced pause (Penh) levels of the MGTG groups (400 mg/kg and 200 mg/kg) were decreased significantly compared with that of control group. The numbers of MGTG groups (400 mg/kg and 200 mg/kg) on lung total cells were decreased significantly compared with that of control group. The numbers of MGTG groups (400 mg/kg and 200 mg/kg) on $CD3^+/CD69^+$, $B220^+/CD22^+$, $B220^+/CD23^+$, $B220^+/lgE^+$, $CCR3^+$ cells were decreased significantly compared with that of control group. The number of MGTG group (400 mg/kg) on $CD3^+/CD49b^+$ cells was decreased significantly compared with that of control group. The level of MGTG groups (400 mg/kg and 200 mg/kg) on IL-4, IL-5, IL-12, TNF-${\alpha}$, OVA-lgE were decreased significantly compared with that of control group. The level of MGTG group (400 mg/kg) on IL-1b, IL-1S, OVA-lgE were decreased significantly compared with that of control group. These results demonstrate that MGTG could be a desirable alternative therapy for allergic asthma by inhibiting the expression of immune cells, the activation of inflammatory mediator.

Multilevel Mediation Analysis: Statistical Methods, Analytic Procedure, and a Real Example (다층자료의 매개효과 분석: 통계방법, 분석절차 및 실례)

  • Park, Sun-Mi;Bak, Byung-Gee
    • Science of Emotion and Sensibility
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    • v.19 no.4
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    • pp.95-110
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    • 2016
  • The purpose of this study was to propose a proper method for the multilevel mediation analysis, for which the hierarchical method should be utilized, then MLM (multilevel modeling) approach as a hierarchical method has been popularly utilized until MSEM (multilevel structural equation modeling) approach was not proposed. This purpose was covered by three research questions about statistical methods, analytic procedure, and real example. First, MSEM statistical method was preferred to MLM method for its estimation accuracy and analytic flexibility. Second, the four-step procedures of model building, assumption examination, model comparison, and coefficient testing were proposed for the multilevel mediation analysis. Third, the real data of 2695 students of elementary and secondary schools and 89 teachers were analyzed in the multilevel directions of $2{\rightarrow}2{\rightarrow}1$ and $1{\rightarrow}1{\rightarrow}2$. Out of these directions of $2{\rightarrow}2{\rightarrow}1$, and $1{\rightarrow}1{\rightarrow}2$ model, only the coefficient of $2{\rightarrow}2{\rightarrow}1$ model was significant at the 95% CI. Mplus programs used for the real example are attached on the Appendix. Based on the results, significance and limitations of this study, were discussed in detail.

Rho-associated Kinase is Involved in Preimplantation Development and Embryonic Compaction in Pigs

  • Son, Myeong-Ju;Park, Jin-Mo;Min, Sung-Hun;Park, Hum-Dai;Koo, Deog-Bon
    • Journal of Embryo Transfer
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    • v.25 no.2
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    • pp.103-110
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    • 2010
  • The first morphogenetic event of preimplantation development, compaction, was required efficient production of porcine embryos in vitro. Compaction of the porcine embryo, which takes place at post 4-cell stage, is dependent upon the adhesion molecule E-cadherin. The E-cadherin through ${\beta}$-catenin contributes to stable cell-cell adhesion. Rho-associated kinase (ROCK) signaling was found to support the integrity of E-cadherin based cell contacts. In this study, we traced the effects of ROCK-1 on early embryonic development and structural integrity of blastocysts in pigs. Then, in order to gain new insights into the process of compaction, we also examined whether ROCK-1 signaling is involved in the regulation of the compaction mediated by E-cadherin of cellular adhesion molecules. As a result, real-time RT-PCR analysis showed that the expression of ROCK-1 mRNA was presented throughout porcine preimplantation stages, but not expressed as consistent levels. Thus, we investigated the blastocyst formation of porcine embryos treated with LPA and Y27632. Blastocysts formation and their qualities in LPA treated group increased significantly compared to those in the Y27632-treated group (p < 0.05). Then, to determine whether ROCK-1 associates embryonic compaction, we explored the effect of activator and/or inhibitor of ROCK-1 on compaction of embryos in pigs. The rate of compacted morula in LPA treated group was increased compared to that in the Y27632-treated group (39.7 vs 12.0%). Furthermore, we investigated the localization and expression pattern of E-cadherin at 4-cell stage porcine embryos in both LPA- and Y27632-treated groups by immunocytochemical analysis and Western blot analysis. The expression of E-cadherin was increased in LPA-treated group compared to that in the Y27632-treated group. The localization of E-cadherin in LPA-treated group was enriched in part of blastomere contacts compared to that Y27632-treated group. ROCK-1 as a crucial mediator of embryo compaction may plays an important role in regulating compaction through E-cadherin of the cell adhesion during the porcine preimplantation embryo. We concluded that ROCK-1 gene may affect the developmental potential of porcine blastocysts through regulating embryonic compaction.

Analysis of the effects of δ-Aminolevulinic acid on the proliferation and apoptosis of mammalian cells (포유류 세포주에서 δ-Aminolevulinic acid (ALA)의 세포증식과 사멸에 미치는 영향분석)

  • Jun, Yong-Woo;Kim, Kun-Hyung;Jo, Su-Yeon;Lee, Jin-A;Jang, Deok-Jin
    • Analytical Science and Technology
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    • v.27 no.5
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    • pp.223-227
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    • 2014
  • ${\delta}$-Aminolevulinic acid (ALA) is a compound which is widely present in the biosphere and plays an important role in the living body as an intermediate of the tetrapyrrole compound biosynthesis pathway that leads to heme in mammals and chlorophyll in plants. ALA is of interest as a biodegradable mediator, a growth regulator, a precursor of heme proteins, and an effective agent used in therapy of cancer. It has been recently reported that ALA is commonly used in dermatology, due to good effects of skin therapy. Although for the last few decades a substantial amount of research has been focused on the elucidation of the mechanism of ALA and the improvement of its therapeutic activity, it's effect on the cell functions and growth was not cleared. Here, we identified that ALA treatment could attenuate cell proliferation of HEK293T and HaCaT cells. In addition, ALA treatement could induce apoptosis of HeLa cells. These results suggest that apoptosis induced by ALA treatment might be responsible for inhibition of cell proliferation. These results propose the possibility of the improved therapeutic strategy making ALA one of the effective drugs used in human cancers.