• Title/Summary/Keyword: BrdU assay

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Cell Cycle Arrest in Human Monocyte Cell Line by Human Cytomegalovirus (인체거대세포바이러스에 의한 인체 단핵구세포의 세포주기 저해)

  • Jang, So-Young;Kim, Mi-Suk;Lee, Chan-Hee
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.299-304
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    • 2008
  • Monocytic cells in myeloid lineage are known for latent site of HCMV Previous studies have suggested that HCMV regulates cell cycle progression in a variety of cells, but studies in monocytic cells are limited. In this study, we attempted to understand cell cycle changes after HCMV infection in the monocytic cell lines. Flow cytometric analyses using propidium iodide revealed that the proportion of G0-G1 phase was increased and the proportion of S phase decreased in HCMV-infected THP-1 cells, but not in HL-60 cells. BrdU-incorporation assay supported that cell proliferation was inhibited in HCMV-infected THP-1 cells by inhibition of de novo DNA synthesis. Western blot analysis revealed that p21, inhibitor of cell cycle progression from G1 phase to S phase, was induced in HCMV-infected THP-1 cells but not in HL-60 cells. Thus, HCMV inhibited cell pro-liferation by arresting the cell cycle at G0-G1 phase through induction of p21 protein in promocytic THP-1 cells.

Acupuncture Suppresses Intrastriatal Hemorrhage-Induced Neuronal Cell Death and Proliferation in Rats

  • Cho, Nam-Hun;Park, Dong-suk;Lee, Jae-Dong
    • The Journal of Korean Medicine
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    • v.24 no.4
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    • pp.127-135
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    • 2003
  • Objectives : Intracerebral hemorrhage (ICH) is one of the most devastating types of stroke. The effect of acupuncture on the intrastriatal hemorrhage-induced neuronal cell death and cell proliferation in rats is examined. Methods : Cell death and cell proliferation in rats was investigated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and immunohistochemistry for caspase-3 and 5-bromo-2'-deoxyuridine (BrdU). Results : Results showed that apoptotic cell death in the striatum and cell proliferation in the hippocampal dentate gyrus significantly increased following intrastriatal hemorrhage in rats, and that acupunctural treatment at the Zusanli acupoint suppressed the hemorrhage-induced increase in apoptosis in the striatum and cell proliferation in the dentate gyrus. Conclusions : It is suggested that acupunctural treatment, especially at the Zusanli acupoint, may aid recovery following central nervous system sequelae following ICH.

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Effects of Ganoderma lucidum on the IL-1, TNF and IL-12 Gene Expression of Macrophages (영지버섯이 탐식세포의 IL-1, TNF 및 IL-12 유전자 발현에 미치는 영향)

  • 배지현
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.5
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    • pp.978-982
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    • 1997
  • In order to investigate the immunomodulatory mechanism of Ganoderma lucidum, the effects of protein-bound polysacchride of Ganoderma lucidum on the proliferation and cytokine gene expression of mouse peritoneal macrophages was studied. In the macrophage proliferation assay using the BrdU labeling reagent, the GLA component extracted from Ganoderma lucidum or GLB from the bud of Ganoderma lucidum were added to the medium at the concentration of 0 to 256ug/ml. DNA synthesis of the macrophage was increased at 16ug/ml of GLA and 64ug/ml of GLB, respectively. In the reverse transcription polymerase chain reaction(RT-PCR), the cytokine(TNF, IL-1, and IL-12) gene and $\beta$-actin expression were also analyzed. 20$\mu\textrm{g}$/ml of either GLA or GLB increased TNF and IL-1 expression of the macrophages.

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Sister Chromatid Exchanges in Lymphocytes on Normal Human Blood Culture with Mercury chloride or Methylmercury Chloride (Mercury chloride 및 Methylmercury chloride가 정상인(正常人)의 혈액배양(血液培養)에서 임파구(淋巴球)의 자매염색분체교환(姉妹染色分體交換)에 미치는 영향(影響))

  • Koh, Dai-Ha
    • Journal of Preventive Medicine and Public Health
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    • v.17 no.1
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    • pp.245-250
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    • 1984
  • Reciprocal exchanges of DNA in sister chromatids (SCEs) are induced by various carcinogens and mutagens, although the quantitative relationship between the number of mutations and SCEs induced varies among chemicals. Nevertheless, the analysis of SCEs production by various agents often proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity. Mercury, even if which has no evidences for mutagenicity and carcinogenicity, is reported to exert some cytotoxic effects, such as chromosomal aberrations or bad influences to ovulation and reproduction in experimental animals, etc.. In this study, tests for sister chromatid exchanges have been carried out on normal human lymphocytes in whole blood culture to add mercury chloride ($HgCl_2$) or methylmercury chloride ($CH_3\;HgCl$) for 72 hr. The results indicate the dose-dependent relationship between the frequencies of SCEs and the concentrations of $HgCl_2,\;CH_{3}HgCl$ and 5-bromo-2'-deoxyuridine (BrdU). Lymphocyte proliferation has depressed in the higher concentration of mercury.

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Stem cell properties of cells derived from canine periodontal ligament (성견 치주인대세포의 줄기세포 특성 연구)

  • Kim, Kyoung-Hwa;Kim, Su-Hwan;Seol, Yang-Jo;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.37 no.3
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    • pp.479-488
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    • 2007
  • In spite of the attention given to the study of mesenchymal stem cells derived periodontal ligament (PDL), there is a lack of information about canine PDL cells. In this study, we characterized canine PDL cells to clarify their stem cell properties, including self renewal, proliferate rate, stem cell markers and multipotency. PDL cells were obtained from extracted premolars of canines, following a colony forming assay and proliferation rate of sub-confluent cultures of cells for self-renewal, immunostaining for STRO-1 and CD146/MUC18 and a differentiation assay for multipotency. Canine PDL cells formed single-cells colonies and 25% of the PDL cells displayed positive staining for BrdU. The cells expressed the mesenchymal stem-cell markers, STRO-1 and CD146/MUC18. Under defined culture conditions, the cells differentiated into osteoblasts and adipocytes, but the cells didn't differentiated into chondrocytes. The findings of this study indicated that the canine PDL cells possess crucial stem cells properties, such as self-renewal and multipotency, and express the mesenchymal stem cell markers on their surface. The isolation and characterization of canine PDL cells makes it feasible to pursue preclinical models of periodontal regeneration in canine.

Angiogenic Inhibitory Effect of Zingiberis Rhizoma (건강의 혈관신생 억제효과에 관한 연구)

  • Nam Sang Choon;Myung Eu gene;Kang Hee;Shim Bum Sang;Kim Sung Hoon;Choi Seung Hoon;Ahn Kyoo Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1608-1612
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    • 2004
  • This study was conducted to investigate angiogenic inhibitory effect of Zingiberis Rhizoma methanol extract using ECV-304 cells and HT1080 fibrosarcoma cells. The viability of ECV-304 was 30% at 50㎍/㎖ of Zingiberis extract and that of HT1080 was 30% at 100㎍/㎖. Using the BrdU incorporation assay, Zingiberis inhibited the DNA synthesis of ECV-304 and HT1080 by 70% and 50% at 200㎍/㎖. In tube formation assay, at 10㎍/㎖ of Zingiberis, tube network began to degrade and at higher doses, it was completely destroyed. Zymography demonstrated that Zingiberis extract decreased MMP-9 at 10㎍/㎖ and higher doses remarkably inhibited the expression of MMP-9. These data indicate that Zingiberis Rhizoma has angiogenic inhibitory effects and shows the possibility of future anti-metastatic drug.

Immatured Type Uterotrophic Assay for Estrogenicity Evaluation of DEHA (미성숙 랫드 자궁비대반응시험을 이용한 DEHA의 내분비계 장애작용 평가)

  • Park, Ki-Dae;Han, Beom-Seok;Jeong, Ja-Young;Oh, Jae-Ho;Cho, Wan-Seob;Cho, Min-Jeong;Choi, Mi-Na;Kim, Sung-Joon;Kim, Seung-Hee
    • Journal of Environmental Health Sciences
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    • v.33 no.5
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    • pp.379-385
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    • 2007
  • This study was aimed to investigate the estrogenic activity of Di-(2-ethylhexyl) adipate (DEHA) using immatured type uterotrophic assay. SD rats were treated with DEHA (40, 200, 1000mg/kg/day), estradiol-3-benzoate (EB) $(1{\mu}g/kg/day)$ as positive control on the assay. In immatured-type uterotrophic assay, relative organ weights of kidney and reproductive organs such as ovary at high-dose group were significantly increased compared to those of vehicle control group. DEHA did not influence the levels of serum FSH and LH, and uterine morphological changes such as luminal epithelial height, myometrial thickness and numbers of uterine gland, and BrdU indices. In these results, there was no significant variation by DEHA treatment, suggesting that DEHA appears not to be a endocrine disrupter with estrogenic activity.

Puerariae Radix Induces Angiogenesis in vitro and in vivo

  • Choi, Do-Young;Kang, Jung-Won;Cho, Eun-Mi;Lee, Jae-Dong;Huh, Jeong-Eun;Yang, Ha-Ru;Baek, Yong-Hyeon;Kim, Deog-Yoon;Cho, Yoon-Je;Kim, Kang-Il;Park, Dong-Suk
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.171-180
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    • 2005
  • Background & Objective : Angiogenesis consists of the proliferation, migration, and differentiation of endothelial cells, and angiogenic factors and matrix protein interactions modulate this process. The aim of this study was to determine whether Puerariae radix could induce angiogenic activity in human umbilical vein endothelial cells (HUVECs). Methods: The angiogenic activity of Puerariae radix were evaluated by using BrdU assay, chemotactic migration assay, tube formation assay, measurement of bFGF in HUVECs, and Matrigel plug assay in mice. Results : Puerariae radix significantly increased HUVECs proliferation in a dose-dependent manner. In addition, Puerariae radix increased migration and tube-like formation in HUVECs. Interestingly,the expression of basic fibroblast growth factor (bFGF), an angiogenesis-stimulating growth factor, was dose-dependently increased by Puerariae radix. The angiogenic activity of Puerariae radix was confirmed using an in vivo Matrigel angiogenesis model, showing promotion of blood vessel formation. Conclusion : Puerariae radix significantly induces angiogenesis in vitro and in vivo. These results suggest that Puerariae radix is a potent angiogenic agent, and a promising drug, for the induction of neovascularization.

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