• Title/Summary/Keyword: BrdU assay

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The Effects of Achyranthis Radix on Short-term Memory and Apoptosis in the Hippocampus of the Gerbil with Transient Global Ischemia (우슬이 뇌허혈 유발 모래쥐의 해마에서 신경세포 사멸과 단기기억력에 미치는 영향)

  • Yoon, Hyun-Seok;Song, Yun-Kyung;Lim, Hyung-Ho
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.2
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    • pp.15-30
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    • 2011
  • Objectives : The present study investigated the effects of Achyranthis Radix on short-term memory, apoptotic neuronal cell death in the hippocampus following transient global ischemia in gerbils. Methods : The gerbils were divided into 5 groups(n=10); Sham operation group, ischemia-induced group, ischemia-induced and 50 mg/kg Achyranthis Radix-treated group, ischemia-induced and 100 mg/kg Achyranthis Radix-treated group, ischemia-induced and 200 mg/kg Achyranthis Radix-treated group. For this study, a step-down avoidance task, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay, immunohistochemistry for caspase-3 and BrdU(5-Bromo-2'-deoxyuridine), and western blotting for bax, bcl-2 were performed. Results : The results revealed that ischemic injury impaired short-term memory and increased apoototic neuronal cell death in the hippocampal CA1(cornu ammonis area 1) region. Ischemic injury enhanced cell proliferation in the hippocampal CA1 region, the compensatory and adaptive process for excessive apoptosis. Achyranthis Radix treatment improved short-term memory by suppressing ischemia-induced apoptotic neuronal cell death in the hippocampal CA1 region. Also, Achyranthis Radix suppressed the ischemia-induced increase in cell proliferation in the hippocampal CA1 region. Conclusions : We showed that Achyranthis Radix alleviates ischemia-induced apoptotic neuronal cell death, thus facilitates the recovery of short-term memory impairment induced by ischemic cerebral injury.

Fabrication of PHBV/Keratin Composite Nanofibrous Mats for Biomedical Applications

  • Yuan, Jiang;Xing, Zhi-Cai;Park, Suk-Woo;Geng, Jia;Kang, Inn-Kyu;Yuan, Jiang;Shen, Jian;Meng, Wan;Shim, Kyoung-Jin;Han, In-Suk;Kim, Jung-Chul
    • Macromolecular Research
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    • v.17 no.11
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    • pp.850-855
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    • 2009
  • Keratin is an important protein used in wound healing and tissue recovery. In this study, keratin was modified chemically with iodoacetic acid (IAA) to enhance its solubility in organic solvent. Poly(hydroxybutylate-co-hydroxyvalerate) (PHBV) and modified keratin were dissolved in hexafluoroisopropanol (HFIP) and electrospun to produce nanofibrous mats. The resulting mats were surface-characterized by ATR-FTIR, field-emission scanning electron microscopy (FE-SEM) and electron spectroscopy for chemical analysis (ESCA). The pure m-keratin mat was cross-linked with glutaraldehyde vapor to make it insoluble in water. The biodegradation test in vitro showed that the mats could be biodegraded by PHB depolymerase and trypsin aqueous solution. The results of the cell adhesion experiment showed that the NIH 3T3 cells adhered more to the PHBV/m-keratin nanofibrous mats than the PHBV film. The BrdU assay showed that the keratin and PHBV/m-keratin nanofibrous mats could accelerate the proliferation of fibroblast cells compared to the PHBV nanofibrous mats.

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.

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.