• Title/Summary/Keyword: thymidine incorporation

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Toxicological Evaluation of Medicinal Plants Used for Herbal Drugs (II) -Acute Toxicity and Effects on DNA Biosynthesis in Bone Marrow Cells and Hemoglobin Content in Blood- (한국산 생약의 약리작용 및 독성연구 (제2보) -급성 독성 및 골수세포의 DNA생합성에 미치는 영향-)

  • Chang, Il-Moo;Kim, Young-Soo;Han, Byung-Hoon
    • Korean Journal of Pharmacognosy
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    • v.13 no.1
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    • pp.14-19
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    • 1982
  • Potential toxicity of 15 medicinal plants used for herbal drugs, which were also described as being tonic for hematopoietic system or being toxic for the system in a oriental book 'Dong Ee Bo Gam', were evaluated in mice. Six plants among 15 plants tested appeared to exhibit acute toxicity along with bone marrow depression or with abnormally enhancing the $^3H-thymidine$ incorporation into DNA biosynthesis in bone marrow cells. Six plants were Paeonia albiflora, Pharbitis nil, Cemphalia lapidescens, Scutellaria baicalensis, Akebia quinata and Glycyrriza uralensis.

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Kaempferol inhibits the platelet-derived growth factor $\beta$-receptor tyrosine-phosphorylation and its downstream intracellular signal transduction pathway in rat aortic vascular smooth muscle cells

  • Kim, Soo-Yeon;Kim, Jin-Ho;Lim, Yong;Yoo, Hwan-Soo;Yun, Yeo-Pyo
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.108.2-108.2
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    • 2003
  • Kaempferol, a flavonol compound, has been reported as the anti-oxidant and anti-angiogenic agent and it has been found to inhibit cell growth in vitro. Abnormal proliferation of vascular smooth muscle cells (VSMCs) plays an important role in development of atherosclerosis. In this study, we examined the anti-proliferative effect and its mechanism on rat aortic VSMCs treated by kaempferol. kaempferol significantly inhibited the platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic VSMCs in concentration-dependent manner by cell count and [$^3$H]-thymidine incorporation assay. (omitted)

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Cloning and Characterization of cDNA for Korean Rockfish (Sebastes schlegeli ) Insulin-like Growth Factor-I

  • Kwon, Mi-Jin;Jo, Jae-Yoon;Nam, Taek-Jeong
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.119-125
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    • 2006
  • To understand the comprehensive mechanisms of biological function for insulin-like growth factor-I (IGF-I) in vertebrates, we have investigated the cDNA sequence of this gene in the korean rockfish (Sebastes schlegeli). The mature form of korean rockfish IGF-I was found to be comprised of 67 amino acid residues, showing about a 7 kDa molecular weight. In this study, we used the polymerase chain reaction (PCR) to obtain a korean rockfish IGF-I (KR IGF-I) cDNA fragment, and methods of rapid amplification of cDNA ends (RACE) to obtain a full length of the KR IGF-I sequence. The KR IGF-I encoded for a predicted amino acid sequence showed identities of 93.6 %, 90.7 %, and 85.4 % in comparison with flounder, chinook salmon, and human IGF-I, respectively. To obtain recombinant biologically active polypeptides, korean rockfish B-C-A-D domains were amplified using the PCR, then the isolated cDNA was expressed in the E. coli BL21(DE3). The recombinant KR IGF-I protein biological function was measured by stimulation of [$^3H$] thymidine incorporation, suggesting the cDNA codes for the korean rockfish proIGF-I.

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Control Mechanisms of Ovarian Follicle Development by Follicle Stimulating Hormone and Pituitary Adenylate Cyclase-activating Polypeptide (난포자극호르몬과 Pituitary Adenylate Cyclase-activating Polypeptide에 의한 난소의 난포성장)

  • Lee, Yu-Il;Shin, Jin-Ok;Kim, Mi-Young;Chun, Sang-Young
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.1
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    • pp.15-23
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    • 2006
  • Objective: Pituitary adenylate cyclase-activating polypeptide (PACAP), a novel hypothalamic neuropeptide, has been suggested to play a role in ovarian folliculogenesis. The present study evaluated the effect of PACAP on the growth of preantral follicles. Methods: Preantral follicles were mechanically isolated from ovaries of 21-day-old rats and cultured in groups for 3 days in serum-free medium in the absence or presence of PACAP-38 ($10^{-6}M$). Results: Treatment with PACAP-38 resulted in an increase in follicle diameter by 75% whereas treatment with follicle stimulating hormone (FSH) increased follicle diameter by 65%. PACAP-38 treatment enhanced the granulosa cell proliferation as measured by thymidine incorporation analysis. Furthermore, the production of progesterone by cultured granulosa cells and GFSHR-17 cell line was stimulated by PACAP-38. Interestingly, PACAP enhanced FSH action on stimulation of SF-1 and aromatase gene expression. Conclusion: The present results demonstrate that PACAP stimulated preantral follicle growth by potentiating proliferation and by stimulating steroidogenesis.

ppGalNAc T1 as a Potential Novel Marker for Human Bladder Cancer

  • Ding, Ming-Xia;Wang, Hai-Feng;Wang, Jian-Song;Zhan, Hui;Zuo, Yi-Gang;Yang, De-Lin;Liu, Jing-Yu;Wang, Wei;Ke, Chang-Xing;Yan, Ru-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5653-5657
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    • 2012
  • Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.

Effect of White Ginseng on the Proliferation of Mouse Peritoneal Macrophages and Their Nitric Oxide Synthesis (백삼성분이 마우스 복강 탐식세포의 증식 및 Nitric Oxide 생성에 미치는 영향)

  • 김주원;배지현
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.4
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    • pp.484-490
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    • 1997
  • In an attempt to investigate the effects of white ginseng on the proliferation and the nitric oxide(NO) secretion of mouse peritoneal macrophages, which are the first mai or defense phagocytes in the immune system, the studies have been carried out. In the macrophage proliferation assay using the $^3$H-thymidine incorporation, the total saponin or Ginsenoside Rb$_2$ were added to the medium at the concentration of 0 to 256$\mu\textrm{g}$/$m\ell$. DNA synthesis of the macrophage was increased at 64$\mu\textrm{g}$/$m\ell$ of total saponin and either 16$\mu\textrm{g}$/$m\ell$ or 64$\mu\textrm{g}$/$m\ell$ of Ginsenoside Rb$_2$, respectively. Also, the effect o(white ginseng on the nitric oxide secretion of the macrophages was investigated. The addition of either total saponin or Ginsenoside Rb$_2$ at the concentration of 20$\mu\textrm{g}$/$m\ell$ significantly increased the secretion of NO from the macrophages. The nitric oxide synthase (NOS) gene expression which is responsible for the synthesis of the nitric oxide has been studied using reverse transcription polymerase chain reaction. In RT-PCR, the $\beta$-actin and nos gene expression have been analyzed. 20$\mu\textrm{g}$/$m\ell$ of either total saponin or Ginsenoside Rb$_2$ increased nos gene expression of the macrophages.

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THE EFFECT OF STATIC MAGNETIC FIELDS ON MOLECULAR AND CELLULAR ACTIVITIES (정자기장이 효소와 세포 활성에 미치는 영향)

  • Park, Jae-Gu;Hwang, Hyeon-Shik
    • The korean journal of orthodontics
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    • v.27 no.6 s.65
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    • pp.929-941
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    • 1997
  • Optimal force for orthodontic treatment is the force that produces a rapid rate of tooth movement without discomfort to the Patient or ensuing tissue damage. Recently considerable interest has been generated in the application of magnets as a way to obtain an optimal force. The purpose of the present study was to investigate the effect of static magnetic fields of Sm-Co magnets on molecular and cellular activities. The distance of erythrocyte sedimentation was measured directly, and the activities and the syntheses of $Fe^{2+}$-related enzymes (catalase and NO synthase) and non $Fe^{2+}$-related enzyme (lactic dehydrogenase) were assayed by the spectrophotometer. The growth and the proliferation of osteoblast-like cells $MC_3T_3-E_1$ were determined by the crystal violet staining and the ${^3}H$-thymidine incorporation. The erythrocytes were exposed to the pole face flux density of 1,400 G (gauss), and the enzymes and osteoblast-like cells $MC_{3}T_3-E_1$ were exposed to the flux density of 7,000 G. The results obtained were as follows: 1. The distance of sedimentation of erythrocyte was not affected by the static magnetic fields. 2. The activities of catalase and lactic dehydrogenase were not affected by the static magnetic fields. 3. The intracellular syntheses of NO synthase and lactic dehydrogenase were not affected by the static magnetic fields. 4. The growth and the proliferation of cultured osteoblast-like cells $MC_{3}T_3-E_1$ were not affected by the static magnetic fields. These results suggested that the molecular and cellular activities were not significantly influenced by the static magnetic fields.

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Wiryeongtang attenuates diabetic renal dysfunction in human renal mesangial cells (위령탕(胃苓湯) 추출물의 사람 유래 신장 메산지움 세포에서의 당뇨병성 신장 손상 개선 효과)

  • Yoon, Jung Joo;Han, Byung Hyuk;Choi, Eun Sik;NamGung, Seung;Jeong, Da Hye;Kim, Hye Yoom;Ahn, You Mee;Lee, Yun Jung;Kang, Dae Gill;Lee, Ho Sub
    • The Korea Journal of Herbology
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    • v.31 no.5
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    • pp.71-78
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    • 2016
  • Objectives : Diabetic nephropathy is one of the most common chronic complications of diabetes and a leading cause of end-stage renal failure in the world. Mesangial cell proliferation is known as the major pathologic features such as glomerulosclerosis and renal fibrosis. Wiryeongtang (WRT) is a well-known traditional herbal formula as therapeutic agents for chronic edema and dysuresia of renal homeostasis. In the present study, we investigated whether WRT inhibits high glucose (HG)-induced renal dysfunction by TGF-β/Smads signal regulation in cultured mesangial cells.Methods : Inhibitory effect of WRT (10-50 ㎍/ml) on HG-stimulated mesangial cells proliferation and dysfunction were evaluated by [3H]-thymidine incorporation, Western blot, and RT-qPCR.Results : WRT significantly decreased HG-accelerated thymidine incorporation in human renal mesangial cell in a dose-dependent levels. WRT induced down-regulation of cyclins/CDKs and up-regulation of CDK inhibitor, p21waf1/cip1 and p27kip1 expression. In addition, HG enhanced expression of dysfunction biomarker such as collagen IV and CTGF, which was markedly attenuated by WRT. WRT decreased TGF-β1 and Smad-2/Smad-4 expression, whereas increased Smad-7 expression under HG. Furthermore, WRT inhibited HG-induced inflammatory factors level such as ICAM-1 and MCP-1 as well as NF-κB p65 nuclear translocation and intracellular ROS production.Conclusions : These results suggested that WRT may alleviate mesangial proliferation and inflammation possibly involved in renal fibrotic process, further diabetic nephropathy through disturbing TGF-β1/Smad signaling and NF-κB/ROS pathway. Thus, WRT might prove to be effective in the treatment of renal dysfunction leading to diabetic nephropathy.

Role of Integrin, FAK (Focal Adhesion Kinase) and ERK (Extracellular Signal Regulated Kinase) on the Suppressed Cell Proliferation of Endometrial Cancer Cells by GnRH (Gonadotropin-Releasing Hormone) (GnRH (Gonadotropin-Releasing Hormone)에 의한 자궁내막암 유래 세포주의 세포 증식 억제 기전에 있어서 Integrin, FAK (Focal Adhesion Kinase) 및 ERK (Extracellular Signal Regulated Kinase)의 역할)

  • Choi, Jong Rak;Park, Dong Wook;Choi, Dong Soon;Min, Churl K.
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.2
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    • pp.115-123
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    • 2006
  • Objective: To investigate new signal transduction cascade through integrin, FAK and ERK in the suppressed cell proliferation by GnRH-I and -II. Method: Human endometrial cancer cells (HEC1A) were cultured under the following condition: DMEM/F12 (10% FBS). GnRH-I and -II were treated time (0, 5, 10, 15, 20, 30 min; 100 nM) and dose (10 nM or 100 nM; 20 min) dependent manner according to experimental purposes. Cell proliferation was measured using [$^3H$] thymidine incorporation assay. Immunoblotting was utilized to detect proteins. Results: GnRH-I and -II inhibited proliferation of HEC1A cells and induced expression of integrin ${\beta}3$. Phosphorylation of FAK and ERK were induced by GnRH-I and -II. Conclusion: GnRH inhibited cell proliferation via the expression of integrin and FAK, ERK phosphorylation.

Influence of Panax Ginseng on Hepatic DNA Synthesis in Mice (고려인삼이 마우스의 간조직 DNA 합성능에 미치는 영향( I ))

  • Chae, Y.B.;Chang, W.S.;Kwon, Y.C.
    • The Korean Journal of Physiology
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    • v.8 no.1
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    • pp.27-30
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    • 1974
  • It was planned to evaluate the influence of Panax Ginseng upon hepatic DNA synthesis in mice by observing incorporation of $[^3H]$ thymidine into the tissue cells. Thirty male mice$(body\;weight:\;18{\sim}20\;g)$ were divided equally into the ginseng and the saline groups. Each animal of the ginseng and the saline groups received every day (subcutaneously) 0.05 m1/10 g body weight of ginseng extract (4mg of ginseng alcohol extract in 1 ml of saline) and the same amount of saline, respectively, for 5 days. On the 5th experimental day, all animals received $1\;{\mu}Ci/g$ body weight of $[^3H]$ thymidine intraperitoneally 2 hours after the last medication. Five animals, at a lime, of each group were sacrificed 1, 10, and 24 hours after thymidine administration, and their hepatic radioactivity was measured autoradiographically in terms of the % number of radioactive cells in 1,000 cell counts (Radioactive Index, R.I.). Following results were obtained: 1. The hepatic radioactive indices obtained from the saline group 1, 10, and 24 hour after $[^3H]$ thymidine administration were $3.23{\pm}0.23,\;5.20{\pm}0.21,\;and\;6.00{\pm}0.30\;(mean{\pm}S.D.)$, respectively. 2. The corresponding values obtained from the ginseng group $(4.22{\pm}0.33,\;6.32{\pm}0.32,\;and\;7.42{\pm}0.35)$ were significant higher than the values of the saline group. The inference from the above results was that the ginseng facilitated hepatic DNA synthesis.

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