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An Experimental Study on the Effect of Low Level Laser and Some Cytokines on Gene Expression of Human Gingival Fibroblasts (저출력레이저조사와 염증성 자극물질이 치은섬유아세포의 유전자 발현에 미치는 영향에 관한 실험적 연구)

  • Jung-Min Kim;Keum-Back Shin
    • Journal of Oral Medicine and Pain
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    • v.19 no.2
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    • pp.57-71
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    • 1994
  • Gingival fibroblasts were cultured and subjected to the test of Northern blot analysis for the demonstration of various mRNA expression in response to the low level laser treatment. For duplication of in vivo. Wound healing process, fibroblasts were pretreated with proinflammatory cytokine interleukin-1$\beta$(IL-1$\beta$) or mitogenic substance phorbol 12-myristate 13-acetate(PMA) prior to laser irradiation. The results were as follows : 1. By the laser irradiation, the gene expression of collagen type I was markedly increased I n gingival fibroblasts, especially in the case of PMA pretreatment. The gene expression of collagen type IV, however, was not only affected by laser irradiation but also by chemical cell stimulation. 2. Oncogene v-myc expression was affected by both laser irradiation and IL-1$\beta$ or PMA stimulation, But v-fos gene expression was not detected in any case of this experimental system. 3. Heat shock gene(Hsp 70)was expressed constiutively, but slightly increased by laser irradiation. 4. mRNA of fibroblast growth factor(FGF) was induced by both laser irradiation and IL-1$\beta$ or PMA treatment.

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Effect of Environmental Enrichment on Cognitive Impairment-induced by Ethanol Exposure in Adolescent Rat

  • Kim, Yoon Ju;Park, Jong Min;Song, Min Kyung;Seong, Ho Hyun;Kim, Youn Jung
    • Journal of Korean Biological Nursing Science
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    • v.18 no.4
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    • pp.274-279
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    • 2016
  • Purpose: Adolescents who experienced the alcohol consumption have gradually increased. Adolescence is a critical period of the neural plasticity in the brain. Neural plasticity is mediated by neurotrophins and has an impact on cognitive function. Environmental enrichment ameliorates the cognitive function and increases neurotrophins. Thus, we investigated the neuroprotective effect of environmental enrichment on ethanol induced cognitive impairment in adolescent rats. Methods: The ethanol groups and the controls groups were injected with ethanol (0.5g/kg) and phosphate buffered saline, respectively, through intraperitoneal from 28th day of birth for 11 days. The environmental enrichment groups were provided larger cages containing toys than the standard cage. Passive avoidance test and Y-maze test were performed to evaluate the spatial memory. Results: Environmental enrichment+ethanol group showed higher alterations than the standard environment+ethanol group in Y-maze test (p<.05). In hippocampus, The environmental enrichment+ethanol group showed significantly higher level of the number of c-fos positive celsl and density of tropomyosin receptors kinase B receptor than the standard environment+ethanol group (p<.05). Conclusion: So, we suggested that the environmental enrichment played a role as a prophylaxis for prevention of memory impairment induced by ethanol exposure in adolescence.

YAC tripeptide of epidermal growth factor promotes the proliferation of HaCaT keratinocytes through activation of EGFR

  • Yoo, Yeon Ho;Kim, Yu Ri;Kim, Min Seo;Lee, Kyoung-Jin;Park, Kyeong Han;Hahn, Jang-Hee
    • BMB Reports
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    • v.47 no.10
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    • pp.581-586
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    • 2014
  • Epidermal growth factor (EGF) is known to play key roles in skin regeneration and wound-healing. Here, we demonstrate that Pep2-YAC, a tripeptide covering residues 29-31 in the B loop of EGF, promotes the proliferation of HaCaT keratinocytes with activity comparable to EGF. The treatment of HaCaT cells with Pep2-YAC induced phosphorylation, internalization, and degradation of EGFR and organization of signaling complexes, which consist of Grb2, Gab1, SHP2, and PI3K. In addition, it stimulated the phosphorylation of ERK1/2 at Thr 202/Tyr 204 and of Akt1 at Ser 473 and the nuclear translocation of EGFR, STAT3, c-Jun, and c-Fos. These results suggest that Pep2-YAC may be useful as a therapeutic agent for skin regeneration and wound-healing as an EGFR agonist.

Effects of Kanghwalsokdan-tang Gamibang Water Extract on Osteoclast Differentiation and Osteoblast Proliferation (강활속단탕가미방(羌活續斷湯加味方)이 파골세포 분화 및 조골세포 활성에 미치는 영향)

  • Jung, Eun-Hye;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.29 no.2
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    • pp.66-82
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    • 2016
  • Objectives : This study was conducted to evaluate the effect of Kanghwalsokdan-tang Gamibang water extract (KSG) on osteoporosis. Methods : RANKL-stimulated RAW 264.7 was used to evaluate inhibitory effect of KSG osteoclast differentiation and gene expression. We counted TRAP (+) multinucleated cells and measured TRAP activity and mRNA expressions of osteoclastogenesis-related genes (NFATc1, MITF, JNK1, cathepsin K, MMP-9) to figure out the effect of KSG on osteoclast. Osteoblastogenesis was also determined in rat calvarial cell. Alkaline phosphatase (ALP) activity, bone matrix protein and collagen synthesis were measured by using murine calvarial cell. Results : KSG inhibited the differentiation of osteoclast precursor cell and expression of genes related osteoclastogenesis like NAFTc1, MITF, c-fos, JNK1, Cathepsin K, MMP-9 and TRAP. KSG increased cell division and function of osteoblast separated from the skull of a rat and ALP synthesis, biosynthesis of bone matrix protein and collagen. Conclusions : Reviewing these results, KSG has efficacy on osteoclast inhibition and osteoblast activation. After further study, KSG will be able to apply for osteoporosis treatment and prevention.

DNA Methylation in Brain and Liver Tissues of Mice Infected with Scrapie Agent (스크래피에 감염된 마우스의 뇌 및 간조직에서의 DNA Methylation)

  • Choi, E.K.;Uyeno, S.;Ono, T.;Carp, R.I.;Kim, Y.S.
    • The Journal of Korean Society of Virology
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    • v.28 no.2
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    • pp.183-192
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    • 1998
  • DNA methylation degree in the several murine brain and liver genes of different ages and after scrapie infection have been examined by using methylation-sensitive restriction endonuclease digestion. We found that the methylation of c-fos and c-myc in the brain and liver was increased during the late fetal to one month postnatal developmental periods. However, those of the SGP-2, $S100{\beta}$, APP950, PrP, and APLP1 genes were decreased at the same periods. The comparison of the DNA methylation patterns between scrapie infected brains and controls demonstrated there is no significant difference in methylation degree of scrapie-infected brains. These observations indicate that DNA methylation might be importantly related to the aging process. The scrapie-infected murine brain was not significantly developed more senescent than the same age-controls did.

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Performance evaluation of smart prefabricated concrete elements

  • Zonta, Daniele;Pozzi, Matteo;Bursi, Oreste S.
    • Smart Structures and Systems
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    • v.3 no.4
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    • pp.475-494
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    • 2007
  • This paper deals with the development of an innovative distributed construction system based on smart prefabricated concrete elements for the real-time condition assessment of civil infrastructure. So far, two reduced-scale prototypes have been produced, each consisting of a $0.2{\times}0.3{\times}5.6$ m RC beam specifically designed for permanent instrumentation with 8 long-gauge Fiber Optic Sensors (FOS) at the lower edge. The sensing system is Fiber Bragg Grating (FBG)-based and can measure finite displacements both static and dynamic with a sample frequency of 625 Hz per channel. The performance of the system underwent validation in the laboratory. The scope of the experiment was to correlate changes in the dynamic response of the beams with different damage scenarios, using a direct modal strain approach. Each specimen was dynamically characterized in the undamaged state and in various damage conditions, simulating different cracking levels and recurrent deterioration scenarios, including cover spalling and corrosion of the reinforcement. The location and the extent of damage are evaluated by calculating damage indices which take account of changes in frequency and in strain-mode-shapes. The outcomes of the experiment demonstrate how the damage distribution detected by the system is fully compatible with the damage extent appraised by inspection.

Characterization of Protein Kinases Activated during Treatment of Cells with Okadaic Acid

  • Bogoyevitch, Marie A.;Thien, Marilyn;Ng, Dominic C.H.
    • BMB Reports
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    • v.34 no.6
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    • pp.517-525
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    • 2001
  • Six renaturable protein kinases that utilize the myelin basic protein (MBP) as a substrate were activated during prolonged exposure of cardiac myocytes to okadaic acid (OA). We characterized the substrate preference and activation of these kinases, with particular emphasis on 3 novel kinases-MBPK-55, MBPK-62 and MBPK-87. The transcription factors c-Jun, Elk, ATF2, and c-Fos that are used to assess mitogen-activated protein kinase activation were all poor substrates for these three kinases. MAPKAPK2 was also not phosphorylated. In contrast, Histone IIIS was phosphorylated by MBPK-55 and MBPK-62. These protein kinases were activated in cultured cardiac fibroblasts, H9c2 cardiac myoblasts, and Cos cells. High concentrations (0.5 to $1\;{\mu}M$) of OA were essential for the activation of the protein kinases in all of the cell types examined, whereas calyculin A [an inhibitor of protein phosphatase 1 (PP1) and PP2A], cyclosporin A (a PP2B inhibitor), and an inactive OA analog all failed to activate these kinases. The high dose of okadaic acid that is required for kinase activation was also required for phosphatase inhibition, as assessed by immunoblotting whole cell lysates with anti-phosphothreonine antibodies. A variety of chemical inhibitors, including PD98059 (MEK-specific), genistein (tyrosine kinase-specific) and Bisindolylmaleimide I (protein kinase C-specific), failed to inhibit the OA activation of these kinases. Thus, MBPK-55 and MBPK-62 are also Histone IIIS kinases that are widely expressed and specifically activated upon exposure to high OA concentrations.

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Platycodon grandiflorum Extracts Exhibits Anti-inflammatory Properties by Down-regulating MAPK Signaling Pathways Lipopolysaccharide-treated RAW264.7 Cells

  • Kim, Hyeon Jin;Jeong, Seong-Yun;Kim, Jin-Kyung
    • Biomedical Science Letters
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    • v.18 no.4
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    • pp.369-376
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    • 2012
  • Platycodon grandiflorum is a medicinal herb that is used to treat pulmonary and respiratory allergic disorders. The objective of this study was to investigate the protective effects of ethyl acetate extract of Platycodon grandiflorum (PGEA) against inflammation and to discern the molecular mechanism of PGEA in lipopolysaccharide (LPS)-induced signal pathways in RAW264.7 macrophage cells. PGEA suppressed the generation of nitric oxide (NO) and the expression of inducible NO synthase induced by LPS in RAW264.7 cells, and inhibited the release of pro-inflammatory cytokines induced by LPS in RAW264.7 cells. Western blot analysis showed that PGEA suppressed LPS-induced phosphorylation of p38 and c-Jun N-terminal kinase (JNK) but not extracellular signal-regulated kinase and $I{\kappa}-B{\alpha}$ degradation. Inactivation of JNK and p38 was effectively alleviated by PGEA, which subsequently affected the activation of c-Jun and c-Fos, which are the essential components of the activator protein-1 (AP-1) transcription complex. Taken together, the results indicate PGEA suppress the activation of p38, JNK, and AP-1, thereby inhibiting the generation of NO and pro-inflammatory cytokines, which affect the regulation of inflammation. PGEA may be useful for the treatment of various inflammatory diseases.

Transcriptional Profile and Cellular Effects on Time Course & Doses Treatment of Methylmercury using Human cDNA Microarray System

  • Kim, Youn-Jung;Yun, Hye-Jung;Kim, Eun-Young;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.176-176
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    • 2003
  • Methylmercury is known to have devastating effects on the mammalian nervous system. When human neuroblastoma SH-SY5Y cells were treated with methylmercury at sublethal concentrations (6.25 uM), up-regulated genes (39) & down-regulated genes (19) were identified by human 8k cDNA microarray. These genes are related with microtubule process, signal transduction pathway and cell death (apoptosis), Apoptosis-associated genes, HSP70, CDK inhibitor 1, FOS-like antigen were up-regulated and microtubule related genes like villin and dynein down-regultaed. To confirm the presence of apoptosis in cultured SH-SY5Y cells treated 6.25 and 1 uM methylmercury, we applied Annexin V-FITC assay followed by flow cytometric measurements after 6 and 24h. Studies on transcriptional and molecular effect by methylmercury may provide an insight into the neurotoxic effects of methylmercury in human neuronal cells and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

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Apoptosis and Anti-proliferaction by Saussurea lappa and Pharbitis nil in AGS Human Gastric Cancer Cell Line

  • Ko Seong-Gyu;Oh Hee-Rah;Lee Sun-Dong;Hwang Gwi-Seo
    • The Journal of Internal Korean Medicine
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    • v.24 no.1
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    • pp.134-143
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    • 2003
  • Objectives : We performed this study to understand the molecular basis of the antitumor effect of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for cancer treatment in Korean traditional medicine. Design: We analyzed, the effect of these medicinal herbs on proliferation and apoptosis of tumor cells and its association with gene expression, We performed semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) analysis of cell cycle- and apoptosis-related genes using a gastric cancer cell line AGS. Results : Cell counting assay and $[^3H]thymidine$ uptake analysis showed that Saussurea lappa and Pharbitis nil strongly inhibit cell proliferation of AGS in a dose-dependent manner. Interestingly, gene espression assay revealed that mRNA espression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly decreased by Saussurea lappa and Pharbitis nil. Furthermore, Saussurea lappa was identified to activate expression of the p53 tumor suppressor and its downstream effector $p21^{Wafl}$, which leads to $G_1$ cell cycle arrest and apoptosis. These observations suggest that the anticancer effect of Saussurea lappa and Pharbitis nil might be associated with their regulatory capability of tumor-related gene expression.

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