• Title/Summary/Keyword: BrdU

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Influence of cold condition exposure on cognitive function and cell proliferation in rats (저온 노출이 인지기능과 뇌신경세포생성에 미치는 영향)

  • Lim, Beak-Vin;Lee, Sung-Pil
    • Science of Emotion and Sensibility
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    • v.14 no.3
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    • pp.425-434
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    • 2011
  • In the present study was to examine the influence of cold stress conditions on memory function in relation with 5-hydroxytryptamine(serotonin, 5-HT), trptophanhydroxylase(TPH) expression and cell proliferation in the hippocampus. For this study, male Sprague-Dawley rats weighing $250{\pm}10g$ (7 weeks in age) were used. The rats were randomly divided into three groups(n = 10 in each group): the $22^{\circ}C$-control group, the $4^{\circ}C$-3 days group, the $4^{\circ}C$-5 weeks group. The environmental temperature at $22^{\circ}C$ set as the normal conditions, $4^{\circ}C$ was as the cold stress conditions. The present results showed that cold stress conditions shorten latency, representing cold stress disturbed memory function. 5-HT and TPH expressions in the dorsal raphe were increased cold stress. Neurogenesis in the dentate gyrus was increased under cold conditions. The present study revealed that cold stress exerted deteriorative memory function. However, through increasing of 5-HT, TPH and BrdU expression under cold stress conditions did not show memory enhancing effect.

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Anti-angiogenic Effects of Shiquandabutang (십전대보탕(十全大補湯)이 혈관신생(血管新生) 억제(抑制)에 미치는 효과(效果))

  • Cui, Xun;Kang, Hee;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok
    • Journal of Korean Traditional Oncology
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    • v.11 no.1
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    • pp.119-134
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    • 2006
  • Shiquandabutang is very famous prescription for tonifying vital energy. We examined the anti-metatstastic effect of Shiquandabutang with in vitro invasion assay model. We performed the following experiments and the results are listed below:Cell viability assay was carried to determine the dose of Shiquandabutang. At lower dose under 200 ${\mu}g/m{\ell}$ (89.6%) viability was very high. But, viability downed as dose grows. At the dose of 600 ${\mu}g/m{\ell}$ (54.2%) viability was almost half of that of control. And at high dose of 1000 ${\mu}g/m{\ell}$ (15.8%) viability was very pure. In BrdU incorporation assay, Shiquandabutang treated groups showed the decreased DNA synthesis rate compared with control group.(200 ${\mu}g/m{\ell}$ (64.4%), 400 ${\mu}g/m{\ell}$ (7.3%)) The results of gelatinase assay showed that Shiquandabutang decreases the gelatinolytic activity of MMP-9. We examined tube formation assay and the result was that Shiquandabutang ihhibits the tube formation at the dose of 200 ${\mu}g/m{\ell}$ and 400 ${\mu}g/m{\ell}$. We examined rat aortic ring assay and the result was that Shiquandabutang ihhibits the angiogenesis of the rat aortic ring at the dose of 400 ${\mu}g/m{\ell}$. From our research, part of the mechanism underlying anti-metastastic effect of Shiquandabutang was proven in vitro. Moreover, we knew that Shiquandabutang is more effectively inhibits the angiogenesis at high dose.

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Knockdown of HMGN5 Expression by RNA Interference Induces Cell Cycle Arrest in Human Lung Cancer Cells

  • Chen, Peng;Wang, Xiu-Li;Ma, Zhong-Sen;Xu, Zhong;Jia, Bo;Ren, Jin;Hu, Yu-Xin;Zhang, Qing-Hua;Ma, Tian-Gang;Yan, Bing-Di;Yan, Qing-Zhu;Li, Yan-Lei;Li, Zhen;Yu, Jin-Yan;Gao, Rong;Fan, Na;Li, Bo;Yang, Jun-Ling
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3223-3228
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    • 2012
  • HMGN5 is a typical member of the HMGN (high mobility group nucleosome-binding protein) family which may function as a nucleosomal binding and transcriptional activating protein. Overexpression of HMGN5 has been observed in several human tumors but its role in tumorigenesis has not been fully clarified. To investigate its significance for human lung cancer progression, we successfully constructed a shRNA expression lentiviral vector in which sense and antisense sequences targeting the human HMGN5 were linked with a 9-nucleotide loop. Inhibitory effects of siRNA on endogenous HMGN5 gene expression and protein synthesis were demonstrated via real-time RT-PCR and western blotting. We found HMGN5 silencing to significantly inhibit A549 and H1299 cell proliferation assessed by MTT, BrdU incorporation and colony formation assays. Furthermore, flow cytometry analysis showed that specific knockdown of HMGN5 slowed down the cell cycle at the G0/G1 phase and decreased the populations of A549 and H1299 cells at the S and G2/M phases. Taken together, these results suggest that HMGN5 is directly involved in regulation cell proliferation in A549 and H1299 cells by influencing signaling pathways involved in cell cycle progression. Thus, our finding suggests that targeting HMGN5 may be an effective strategy for human lung cancer treatment.

Inhibitory Effect of YBR on Hepatic Fibrogenesis (YBR의 간섬유화(肝纖維化)억제 효과(效果)에 관한 연구(硏究))

  • Seung, Hyun-Seok;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.314-330
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    • 2010
  • Objective : This study was performed to investigate the anti-fibrogenic effect and changes of inflammation-related genes by YBR I and YBR II (YBR I: Arteisiae Capillaris Herba, Atractylodis Rhizoma Alba, Hoelen/ YBR II: YBR I +Sanguisorbae Radix, Biotae Cacumen, Cirsii Japonici Herba) on HSC(hepatic stellate cells)-T6 and TAA-induced rat liver tissue. Materials and Methods : HSC-T6 were treated with various concentrations of distilled-water extract YBR I and YBR II extract for 24, 48 and 72 hours. After the treatment, cell viability, proliferation, procollagen levels and IL-6 levels were measured by using MTT Assay, BrdU Assay, Procollagen Type 1 C-peptide EIA kit, and Murine IL-6 ELISA Development kit. Rat liver fibrosis was induced by intraperitoneal TAA injection of 150mg/kg 3 times a week for 6 weeks. After the treatment, body weight, liver & spleen weights, liver function test, complete blood cell count and change of portal pressure were studied. In addition, gene expressions of ASMA, IL-6, MMP-2, TIMP-1 and TIMP-2, all of which are known to be associated with liver fibrosis, were analyzed by using Real-Time PCR. After YBR I and YBR IItreatment, percentages of collagen in TAA-induced rat liver tissue were measured. Results : The viability and proliferation of the HSC-T6 decreased as the concentration increased. The production of procollagen decreased as the concentration increased. The production of IL-6 was little influenced by YBR I and YBR II. There was no difference in rat body weight between the TAA-only group and the YBR groups. Compared with rat liver weight of TAA-only group, that of the YBR groups increased. In the YBR I group, the serum level of AST elevated by TAA injection significantly decreased and in the YBR I and II group, the serum level of ALP and ALT elevated by TAA injection decreased. In the YBR I group, white blood cell count elevated by TAA injection decreased but platelets increased. In the YBR I group, the portal pressure elevated by TAA injection significantly decreased. Decreases in the gene expression of ASMA and MMP-2 were observed in the YBR I group. The gene expression of IL-6 was little influenced by YBR I and YBR II -treated groups. In the histological finding, TAA injections caused severe fibrosis, but YBR I and YBR II treatment significantly reduced the amounts of hepatic collagens. Conclusions : These results suggest that YBR I and II have inhibitory effects on the hepatic fibrogenesis.

Spherical and Aspherical RGP Lens Fitting to Epithelium and Endothelium of Rabbit's Cornea -Scanning Electron Microscopy (구면과 비구면 RGP 렌즈 장기 착용시 각막 상피, 내피에 미치는 영향-주사전자현미경적 관찰)

  • Kim, In-Suk;Ryu, Gun-Chang;Chae, Soo-Chul;Jeon, Chang-Jin
    • Applied Microscopy
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    • v.36 no.3
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    • pp.227-234
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    • 2006
  • To investigate the comparative effect of spherical and aspherical RGP lens were worn during 3 weeks on rabbit's cornea. Four white rabbits were worn right eyes with spherical lens and 4 white rabbits were worn right eyes with aspherical RGP lens. Left eyes were served as control. The rabbits were sacrificed at 3 weeks after fitting and observed morphological changes by scanning electron microscopy and also investigate proliferation rate of the corneal epithelium with RGP wearing. After spherical RGP lens wearing, the epithet layer damaged compared to aspherical lens. The superficial cell layer strip off seriously, cell size significantly changed abnormal. Both spherical and aspherical RGP lens fitting group showed so many bacteria and back surface of lens was found like a fern shape. The aspherical RGP lens original material type was some formal than spherical lens. We thought that these pattern was significantly altered with spherical lens by prohibited transmitter oxygen from atmosphere therefore the epithelium shape was changed. This suggested wearing the aspherical lens might be less physiologic than shperical lens fitting.

THE EFFECT OF DIFFERENTIAL MODULATION OF N-METHYL-D-ASPARTATE RECEPTOR ON THE PROLIFERATION OF PRIMARY CULTURED NORMAL HUMAN ORAL KERATINOCYTES: DNA SYNTHESIS RATE ANALYSIS (N-methyl-D-aspartate 수용기의 다양한 조절이 일차 배양된 정상사람구강각화세포의 증식에 미치는 영향; DNA 합성율 평가)

  • Kim, In-Soo;Paik, Ki-Suk;Chang, Mi-Sook;Lee, Won;Lee, Seung-Pyo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.2
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    • pp.124-130
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    • 2007
  • In the present study, I investigated the effects of N-methyl-D-aspartate (NMDA), arachidonic acid (AA), and Nitric Oxide Synthase Inhibitor (NOSI), alone or in combination, on the proliferation of cultured primary normal human oral keratinocytes (NHOK). The purpose of this study was therefore the preliminary study for the examination of the interaction between these agents and NHOK in order to elucidate the mechanisms by which epithelial growth and regeneration are regulated. NHOK were obtained from gingival tissue of 20 individuals aged 20 to 29, and third passage (P3) cells were used for this study. The DNA synthesis was measured by the BrdU assay. Addition of low concentration of AA ($1{\mu}M$) and high concentration of AA with NMDA group (NMDA+AA $10{\mu}M$) made DNA synthesis rate increase significantly at the early stage. Adding NNA ($10{\mu}M$) affected DNA synthesis rate to increase significantly in 4 hours. At the early stage, DNA synthesis was significantly active in the NOS-I with NMDA groups than in the control and the NMDA-only group, while it didn't become statistically meaningful in 24 hours. AA $1{\mu}M$ and NNA $10{\mu}M$ may induce the proliferation of the NHOK independently and NOS-I may induce the proliferation of the NHOK with NMDA. These reactions might be related to the NMDA receptor in the cell and the change of the intracellular calcium ion concentration.

Differential Anti-Carcinogenic Effect of Mountain Cultivated Ginseng and Ginseng on Mouse Skin Carcinogenesis (마우스 피부암에 대한 장뇌삼과 인삼의 특이적 항암 효능)

  • Lee, Min-Hee;Choi, Sang-Won;Kim, Eun-Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.4
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    • pp.462-470
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    • 2012
  • Mountain cultivated ginseng (MCG) is a type of Panax ginseng C. A. Mayer, grown in the mountains by artificial seeding. In general, it has been known that the biophysical activities of MCG is greater than that of ginseng. However, the in vivo efficacy of MCG on cancer has not been studied. In this study, we investigated the anti-carcinogenic effect of MCG and ginseng using the 7,12-dimethylbenz[a]anthracene (DMBA)-12-O-tetradecanoylphorbol- 13-acetate (TPA) two stage mouse skin carcinogenesis model. Six weeks of female ICR mice were divided into control, MCG, and ginseng diet groups and were subjected into two different experimental protocols. In the first study, each experimental diet was fed with TPA promotion for 24 weeks. The result showed that supplementation of MCG reduced tumor incidence, tumor multiplicity, and tumor size compared to those of the control and ginseng groups. In the second study, 3 groups of mice were supplied with each diet 4 weeks before DMBA tumor initiation, until the end of experiment. The result showed that tumor incidence, tumor multiplicity, and tumor size were reduced in the ginseng diet group compared to those of the control and MCG groups. TPA-induced BrdU incorporation was also significantly reduced in the ginseng diet group. Taken together, these results suggest that MCG is chemotherapeutic, whereas ginseng has a chemopreventative effect on mouse skin cancer.

Effects of a gintonin-enriched fraction on hair growth: an in vitro and in vivo study

  • Lee, Na-Eun;Park, Sang-Deuk;Hwang, Hongik;Choi, Sun-Hye;Lee, Ra Mi;Nam, Sung Min;Choi, Jong Hee;Rhim, Hyewhon;Cho, Ik-Hyun;Kim, Hyoung-Chun;Hwang, Sung-Hee;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.168-177
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    • 2020
  • Background: Ginseng has been widely used as a health-promoting tonic. Gintonin present in ginseng acts as a lysophosphatidic acid (LPA) receptor ligand that activates six LPA receptor subtypes. The LPA6 subtype plays a key role in normal hair growth, and mutations in the LPA6 receptor impair normal human hair growth. Currently, human hair loss and alopecia are concerning issues that affect peoples' social and day-to-day lives. Objective: We investigated the in vitro and in vivo effects of a gintonin-enriched fraction (GEF) on mouse hair growth. Methods: Human hair follicle dermal papilla cells (HFDPCs) and six-week-old male C57BL/6 mice were used. The mice were divided into the four groups: control, 1% minoxidil, 0.75% GEF, and 1.5% GEF. The dorsal hair was removed to synchronize the telogen phase. Each group was treated topically, once a day, for 15 days. We analyzed hair growth activity and histological changes. Results: GEF induced transient [Ca2+]i, which stimulated HFDPC proliferation and caused 5-bromo-2'-deoxyuridine (BrdU) incorporation in a concentration-dependent manner. GEF-mediated HFDPC proliferation was blocked by the LPA receptor antagonist and Ca2+ chelator. HFDPC treatment with GEF stimulated vascular endothelial growth factor release. Topical application of GEF and minoxidil promoted hair growth in a dose-dependent manner. Histological analysis showed that GEF and minoxidil increased the number of hair follicles and hair weight. Conclusion: Topical application of GEF promotes mouse hair growth through HFDPC proliferation. GEF could be one of the main components of ginseng that promote hair growth and could be used to treat human alopecia.

20(S)-protopanaxadiol and oleanolic acid ameliorate cognitive deficits in APP/PS1 transgenic mice by enhancing hippocampal neurogenesis

  • Lin, Kaili;Sze, Stephen Cho-Wing;Liu, Bin;Zhang, Zhang;Zhang, Zhu;Zhu, Peili;Wang, Ying;Deng, Qiudi;Yung, Ken Kin-Lam;Zhang, Shiqing
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.325-333
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    • 2021
  • Background: Alzheimer's disease (AD) is one of the most prevalent neurodegenerative disorders. Enhancing hippocampal neurogenesis by promoting proliferation and differentiation of neural stem cells (NSCs) is a promising therapeutic strategy for AD. 20(S)-protopanaxadiol (PPD) and oleanolic acid (OA) are small, bioactive compounds found in ginseng that can promote NSC proliferation and neural differentiation in vitro. However, it is currently unknown whether PPD or OA can attenuate cognitive deficits by enhancing hippocampal neurogenesis in vivo in a transgenic APP/PS1 AD mouse model. Here, we administered PPD or OA to APP/PS1 mice and monitored the effects on cognition and hippocampal neurogenesis. Methods: We used the Morris water maze, Y maze, and open field tests to compare the cognitive capacities of treated and untreated APP/PS1 mice. We investigated hippocampal neurogenesis using Nissl staining and BrdU/NeuN double labeling. NSC proliferation was quantified by Sox2 labeling of the hippocampal dentate gyrus. We used western blotting to determine the effects of PPD and OA on Wnt/GSK3β/β-catenin pathway activation in the hippocampus. Results: Both PPD and OA significantly ameliorated the cognitive impairments observed in untreated APP/PS1 mice. Furthermore, PPD and OA significantly promoted hippocampal neurogenesis and NSC proliferation. At the mechanistic level, PPD and OA treatments resulted in Wnt/GSK-3β/β-catenin pathway activation in the hippocampus. Conclusion: PPD and OA ameliorate cognitive deficits in APP/PS1 mice by enhancing hippocampal neurogenesis, achieved by stimulating the Wnt/GSK-3β/β-catenin pathway. As such, PPD and OA are promising novel therapeutic agents for the treatment of AD and other neurodegenerative diseases.

Transforming growth factor-β promoted vascular endothelial growth factor release by human lung fibroblasts (인간 폐섬유아세포에서 TGF-β 자극에 의한 VEGF 분비)

  • Park, Sang-Uk;Shin, Joo-Hwa;Shim, Jae-Won;Kim, Deok-Soo;Jung, Hye-Lim;Park, Moon-Soo;Shim, Jung-Yeon
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.879-885
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    • 2008
  • Purpose : The human lung fibroblast may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, which are important in airway remodeling. Vascular endothelial growth factor (VEGF) induces mucosal edema and angiogenesis. Thymus and activation regulated chemokine (TARC) induces selective migration of T helper 2 cells. We investigated whether human lung fibroblasts produced VEGF and TARC, and the effects were augmented with the co-culture of fibroblasts and human bronchial smooth muscle cells (HBSMC), and whether dexamethasone can inhibit the proliferation and the release of VEGF in lung fibroblasts. Methods : Human lung fibroblasts were cultured with and without HBSMC, growth-arrested in serum-deprived medium, and pretreated with dexamethasone for 16 hours. After 24-hour stimulation with platelet derived growth factor-BB (PDGF-BB) and/or transforming growth factor-${\beta}$ (TGF-${\beta}$), culture supernatant was harvested for assays of VEGF and TARC. Cell proliferation was assayed using BrdU cell proliferation ELISA kit. Results : 1) The release of VEGF was significantly increased after stimulation with TGF-${\beta}$, and its release was augmented when co-stimulated with PDGF and TGF-${\beta}$. 2) VEGF release induced by PDGF or TGF-${\beta}$ was inhibited by dexamethasone. 3) There was no synergistic effect on the release of VEGF when human lung fibroblasts were co-cultured with HBSMC. 4) Dexamethasone did not suppress human lung fibroblasts proliferations. 5) Neither TGF-${\beta}$ nor PDGF induced TARC release from lung fibroblasts. Conclusion : Human lung fibroblasts may modulate airway remodeling by release of VEGF, but they have no synergistic effects when co-cultured with HBSMC. Dexamethasone suppresses VEGF release, not proliferation of lung fibroblast.