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Effect of Moutan Cortex Radicis on gene expression profile of differentiated PC12 rat cells oxidative-stressed with hydrogen peroxide (모단피의 PC12 cell 산화억제 효과 및 neuronal 유전자 발현 profile 분석에 대한 연구)

  • Kim Hyun Hee;Rho Sam Woong;Na Youn Gin;Bae Hyun Su;Shin Min Kyu;Kim Chung Suk;Hong Moo Chang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.529-541
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    • 2003
  • Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

Identification of Glycine max Genes Expressed in Response to Soybean mosaic virus Infection

  • Jeong, Rae-Dong;Lim, Won-Seok;Kwon, Sang-Wook;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.21 no.1
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    • pp.47-54
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    • 2005
  • Identification of host genes involved in disease progresses and/or defense responses is one of the most critical steps leading to the elucidation of disease resistance mechanisms in plants. Soybean mosaic virus (SMV) is one of the most prevalent pathogen of soybean (Glycine max). Although the soybeans are placed one of many important crops, relatively little is known about defense mechanism. In order to obtain host genes involved in SMV disease progress and host defense especially for virus resistance, two different cloning strategies (DD RT-PCR and Subtractive hybridization) were employed to identify pathogenesis- and defenserelated genes (PRs and DRs) from susceptible (Geumjeong 1) and resistant (Geumjeong 2) cultivars against SMV strain G7H. Using these approaches, we obtained 570 genes that expressed differentially during SMV infection processes. Based upon sequence analyses, differentially expressed host genes were classified into five groups, i.e. metabolism, genetic information processing, environmental information processing, cellular processes and unclassified group. A total of 11 differentially expressed genes including protein kinase, transcription factor, other potential signaling components and resistant-like gene involved in host defense response were selected to further characterize and determine expression profiles of each selected gene. Functional characterization of these genes will likely facilitate the elucidation of defense signal transduction and biological function in SMV-infected soybean plants.

The Impacts of Social Support and Psychological Factors on Guild Members' Flow and Loyalty in MMORPG (MMORPG에서 길드 구성원들의 사회적 지지와 심리적 요인들이 플로우 및 충성도에 미치는 영향)

  • Kang, Ju-Seon;Ko, Yoon-Jung;Ko, Il-Sang
    • Asia pacific journal of information systems
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    • v.19 no.3
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    • pp.69-98
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    • 2009
  • We investigated what factors motivate gamers to participate in a guild and why they continue to be engaged as members of the guild. We find that, based on the result of focus group interviews with MMORPG gamers, social support and self-esteem factors play important roles. Considering both prior research and the focus group interviews we have conducted, we define social support and character control as independent variables. Character identity, guild identity, and self-esteem are proposed as mediating variables while guild flow and game loyalty as dependent variables. Accordingly, we develop the research model and hypotheses, and verify them empirically. Based on our experiences of playing the WoW game, we proposed a research model and conducted focus-group interviews (FGIs). FGIs involve formulating a hypothesis and then collecting some relevant data. FGIs were conducted face-to-face with students of C University in Korea. We formulated structured interview schedules, and the questions were based on our research variables and personal experiences. The questions for the interviews encompassed the following areas: (a) the demographic characteristics of the focus group; (b) the number of years for which respondents had played online games; (c) the motive for starting a game; (d) the number of game-characters assumed by each gamer; (e) the type of game played; and (f) other issues such as the reasons for involvement in the play, the willingness to reuse the game in case new versions were released, etc. On average, it took two hours to interview each of three groups. A primary set of FGIs was conducted with three groups on the premise that there would be some differences caused by character race (Horde vs. Alliance) or by playable server (Normal vs. Combat). With respect to the manner of playing, we found that guild members shared information, felt a sense of belonging, and played computer games for quite a long time through the guild; however, they did not undergo these experiences when playing alone. Gamers who belonged to a specific guild helped other players without expecting compensation for that, freely shared information about the game, gave away items for free, and more generous with other members who made mistakes. The guild members were aware of the existence other members and experienced a sense of belonging through interactions with, and evaluations from, other players. It was clear that social support was shown within the guild and that it played an important role as a major research variable. Based on the results of the first FGIs, a second set of in-depth FGIs was carried out with a focus on the psychology of the individual within the guild and the social community of the guild. The second set of FGIs also focused on the guild's offline meetings. Gamers, over all, recognize the necessity of joining a community, not only off-line but also online world of the guild. They admit that the guild is important for them to easily and conveniently enjoy playing online computer games. The active behavior and positive attitudes of existing guild members can motivate new members of the guild to adapt themselves to the guild environment. They then adopt the same behaviors and attitudes of established guild members. In this manner, the new members of the guild strengthen the bonds with other gamers while feeling a sense of belonging, and developing social identity, thereby. It was discovered that the interaction among guild members and the social support encouraged new gamers to quickly develop a sense of social identity and increase their self-esteem. The guild seemed to play the role of socializing gamers. Sometimes, even in the real world, the guild members helped one another; therefore, the features of the guild also spilled over to the offline environment. We intend to use self-esteem, which was found through the second set of FGIs, as an important research variable. To collect data, an online survey was designed with a questionnaire to be completed by WoW gamers, who belong to a guild. The survey was registered on the best three domestic game-sites: 'WoW playforum,' 'WoW gamemeca,' and 'Wow invent.' The selected items to be measured in the questionnaire were decided based on prior research and data from FGIs. To verify the content of the questionnaire, we carried out a pilot test with the same participants to point out ambiguous questions as a way to ensure maximum accuracy of the survey result. A total of 244 responses were analyzed from the 250 completed questionnaires. The SEM analysis was used to test goodness-of-fit of the model. As a result, we found important results as follows: First, according to the statistics, social support had statistically significant impacts on character control, character identity, guild identity and self-esteem. Second, character control had significant effects on character identity, guild identity and self-esteem. Third, character identity shows its clear impact on self-esteem and game loyalty. Fourth, guild identity affected self-esteem, guild flow and game loyalty. Fifth, self-esteem had a positive influence on the guild flow. These days, the number of virtual community is rising along with its significance largely because of the nature of the online games. Accordingly, this study is designed to clarify the psychological relationship between gamers within the guild that has been generally established by gamers to play online games together. This study focuses on the relationships in which social support influences guild flow or game loyalty through character control, character identity, guild identity, and self-esteem, which are present within a guild in the MMORPG game environment. The study results are as follows. First, the effects of social support on character control, character identity, guild identity and self-esteem are proven to be statistically significant. It was found that character control improves character identity, guild identity and self-esteem. Among the seven variables, social support, which is derived from FGIs, plays an important role in this study. With the active support of other guild members, gamers can improve their ability to develop good characters and to control them. Second, character identity has a positive effect on self-esteem and game loyalty, while guild identity has a significant effect on self-esteem, guild flow and game loyalty. Self-esteem affects guild flow. It was found that the higher the character and guild identities become, the greater the self-esteem is established. Contrary to the findings of prior research, our study results indicate that the relationship between character identity and guild flow is not significant. Rather, it was found that character identity directly affects game players' loyalty. Even though the character identity had no direct effect on increasing guild flow, it has indirectly affected guild flow through self-esteem. The significant relationship between self-esteem and guild flow indicates that gamers achieve flow, i.e., a feeling of pleasure and excitement through social support. Several important implications of this study should be noted. First, both qualitative and quantitative methods were used to conduct this study. Through FGIs, it was observed that both social support and self-esteem are important variables. Second, because guilds had been rarely studied, this research is expected to play an important role in the online community. Third, according to the result, six hypotheses (H1, H5, H6, H7, H8, and H11) setup based on FGIs, were statistically significant; thus, we can suggest the corresponding relationships among the variables as a guideline for follow-up research. Our research is significant as it has following implications: first, the social support of the guild members is important when establishing character control, character identity, guildidentity and self-esteem. It is also a major variable that affects guild flow and game loyalty. Second, character control when improved by social support shows notable influence on the development of character identity, guild identity and self-esteem. Third, character identity and guild identity are major factors to help establish gamers' own self-esteem. Fourth, character identity affects guild flow through self-esteem and game loyalty. The gamers usually express themselves through characters; the higher character identity is, the more loyalty a gamer has. Fifth, guild identity, established within the guild, has clear effects on self-esteem, guild flow and game loyalty. Sixth, qualitative and quantitative methods are employed to conduct this study. Based on the results of focus group interviews and SEM analysis, we find that the social support by guild members and psychological factors are significant in strengthening the flow of guild and loyalty to the game. As such, game developers should provide some extra functions for guild community, through which gamers can play online games in collaboration with one another. Also, we suggest that positive self-esteem which is built up through social support can help gamers achieve higher level of flow and satisfaction, which will consequently contribute to minimizing the possibility for the players to develop negative attitude toward the guild they belong to.