• Title/Summary/Keyword: [$^3H$]-thymidine

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Effect of Glycyrrhizae Radix on the Immune Responses(I) - Immuno-regulatory Action of 50% Methanol Extract - (감초가 면역반응에 미치는 영향 (I) - 50% 메탄올 엑스의 면역조절작용 -)

  • 한종현;오찬호;은재순
    • YAKHAK HOEJI
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    • v.35 no.3
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    • pp.154-164
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    • 1991
  • These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

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Expression of the Functional Recombinant Interleukin-16 in E. coli and Mammalian Cell Lines

  • Kim, Seon-Young;Lee, Chang-Hun;Kim, Kyung-Joo;Kim, Yeon-Soo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.234-241
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    • 2001
  • The C-terminal 393 bp region of the human interleukin-16 (IL-16) gene was cloned and expressed in E. coli along with mammalian cell lines. Recombinant IL-16 expressed from E. coli was 22 kDa on SDS-PAGE and showed 260% of chemoattractant activity at a concentration of $0.1\;{\mu}g/ml$. HeLa, COS, and Neuro-2a cells were transduced by recombinant retrovirus vector pLNC/IL-16/IRES/TK and the intracellular and secreted amounts of IL-16 produced by HeLa/IL-16/TK, COS/IL-16/TK, and Neuro-2a/IL-16/TK cells were determined by enzyme-linked immunosorbent assay (ELISA). HeLa/IL-16/TK $(1{\times}10^5)$ and COS/IL-16/TK $(1{\times}10^5)$ cells secreted 36.1 and 13.3 ng of IL-16 for 48 h, respectively. Forty-nine ng and 86.4 ng of IL-16 remained in the cell lysates of HeLa/IL-16/TK and COS/IL-16/TK. Intracellular and secreted amounts of IL-16 from Neuro-2a/IL-16/TK $(5{\times}10^5)$ cells during 24 h cultivation were 50 ng and 3.3 ng, respectively. Also, HeLa and COS cells wee stably transfected with mammalian expression vector pCRIII/IL-16. Both culture media and cell lysates prepared from HeLa/IL-16 cells and COS/IL-16 cells showed chemoattractant activity ranging from 190% to 460% as compared to the control experiment. Expression of the herpes simplex virus thymidine kinase (HSV0tk) gene in pLNC/IL-16/ IRES/TK bicistronic retroviral expression vector was verified by performing a genciclovir (GCV) sensitivity assay. Finally, IL-16 repressed Tat-transactivated human immunodeficiency virus type 1 long terminal repeat (HIV-1 LTR) promoter activity.

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Drug Resistance Effects of Ribosomal Protein L24 Overexpression in Hepatocellular Carcinoma HepG2 Cells

  • Guo, Yong-Li;Kong, Qing-Sheng;Liu, Hong-Sheng;Tan, Wen-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9853-9857
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    • 2014
  • Background: The morbidity and mortality rate of liver cancer continues to rise in China and advanced cases respond poorly to chemotherapy. Ribosomal protein L24 has been reported to be a potential therapeutic target whose depletion or acetylation inhibits polysome assembly and cell growth of cancer. Materials and Methods: Total RNA of cultured amycin-resistant and susceptible HepG2 cells was isolated, and real time quantitative RT-PCR were used to indicate differences between amycin-resistant and susceptible strains of HepG2 cells. Viability assays were used to determine amycin resistance in RPL24 transfected and control vector and null-transfected HepG2 cell lines. Results: The ribosomal protein L24 transcription level was 7.7 times higher in the drug-resistant HepG2 cells as compared to susceptible cells on quantitative RT-PCR analysis. This was associated with enhanced drug resistance as determined by methyl tritiated thymidine (3H-TdR) incorporation. Conclusions: The ribosomal protein L24 gene may have effects on drug resistance mechanisms in hepatocellular carcinoma HepG2 cells.

Abundance and Structure of Microbial Loop Components (Bacteria and Protists) in Lakes of Different Trophic Status

  • Chrost, Ryszard J.;Tomasz, Adamczewski;Kalinowska, Krystyna;Skowronska, Agnieszka
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.858-868
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    • 2009
  • The abundance, biomass, size distribution, and taxonomic composition of bacterial and protistan (heterotrophic and autotrophic nanoflagellates and ciliates) communities were investigated in six lakes of Masurian Lake District (north-eastern Poland) differing in trophic state. Samples were taken from the trophogenic water layer during summer stratification periods. Image analysis techniques with fluorescent in situ hybridization (FISH) as well as [$^3H$]-methyl-thymidine incorporation methods were applied to analyze differences in the composition and activity of bacterial communities. The greatest differences in trophic parameters were found between the humic lake and remaining non-humic ones. The same bacterial and heterotrophic nanoflagellate (HNF) cell size classes dominated in all the studied lakes. However, distinct increases in the contributions of large bacterial (>$1.0{\mu}m$) and HNF (>$10{\mu}m$) cells were observed in eutrophic lakes. The bacterial community was dominated by the ${\beta}$-Proteohacteria group, which accounted for 27% of total DAPI counts. Ciliate communities were largely composed of Oligotrichida. Positive correlations between bacteria and protists, as well as between nanoflagellates (both heterotrophic and autotrophic) and ciliates, suggest that concentrations of food sources may be important in determining the abundance of protists in the studied lakes.

The Effect of Cyclosporin A on the Growth of human Glioma Cell Lines

  • Pyen, Jhin-Soo;Kim, Soo-Kie;Choi, Sun-Ju;Park, Yoon-Sun;Cho, Hyun-Chul;Han, Young-Pyo
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.379-383
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    • 1997
  • Cyclosporin A, an potent immunosuppressant, has been known to be one of the modulators of drug resistance as well as a cytostatic drug. Despite many attempts to basic or clinical application of cyclosporin A, there are few reports on the inhibition of brain tumor cells. In the present experiment, the possibility of cyclosporin A as synergic adjuvant was investigated by MTT assay, $[^{3}H]$ thymidine uptake and through flowcytometric anaysis. Sole treatment of cyclosporin A on the CRT and CH235-MG glioma cell line revealed dose dependent cytotoxicity within a range of tested dose. Combined treatment of cyclosporin A with ACNU, BCNU and hydroxyurea on various glioma cancer cell line led to a significant synergistic cytotoxicity as well as inhibition of DNA synthesis with dose-dependency. In addition, cyclosporin A alone or combined treatment caused discernible changes of cell cycle in the tested cells. These data provide that cyclosporin A could potentiate the effect of nitrosourea compounds in vitro on human glioma cells.

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RADIOAUTOGRAPHIC ANALYSIS OF CHANGES IN DIFFERENT PHASES OF CELL KINETICS IN MURINE ORAL MUCOSA

  • PARK CHANG SUCK;You Dong Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.13 no.1
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    • pp.29-73
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    • 1983
  • The age related changes in the life cycle of the progenitor cell population of murine oral epithelia was studied. Using radioautographic methods which have been adopted in previous cell cycle studies, the age-related changes of different phases in renewing cells of the palatal, buccal and lingual mucosae were determined. The results confirm published findings on cell cycle changes of epithelia with aging and illustrated further that mitotic phase which has hither to been considered stationary, also changes with aging. The major parts revealed by this study are as follows: 1) The basal progenitor cells in different regions of oral mucosa have different generation times. 2) The basal cell cycle time increases as a function of aging and the region most affected by aging appears to be the epithelium of the cheek. 3) The phases of the cell cycle affected by the process of aging are in increasing order of magnitude: M-, S- and G₁-phase. 4) The age elated change in the number of DNA synthesizing basal progenitor cells occurs at two age periods. Between 1 and 12 months of life it decreases, while from 12 to 20 months it increases.

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The Effects of Saussurea Radix and Plantaginis Herba on Cellular Viability, Proliferation, Apoptosis and Expression of Cell Cycle-related Genes in Gastric Cancer Cells (목향(木香)과 차전초(車前草)가 위암세포(胃癌細胞)의 활성(活性), 증식(增殖), 자기살해능(自己殺害能) 및 세포주기관련 유전자 발현에 미치는 영향)

  • Oh, Hee-Rah;Ko, Seong-Gyu
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.7 no.1
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    • pp.1-18
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    • 2001
  • Objective: This experimental study was carried out to evaluate the effects of Saussurea Radix and Plantaginis Herba on cellular viability, proliferation, apoptosis and expression of the cell cycle-related genes in cultured gastric cancer cells. Method :MTT assay for analysis of cellular toxicity and the effect on suppression of cellular viability, $[^{3}H]$ thymidine incorporation assay for evaluation of the effect on suppression of DNA replication, tryphan blue exclusion assay for measurement of induction of apoptosis and Quantitative RT-PCR for analysis of the effects on expression of cell cycle or apoptosis-related genes were performed. Results: Antitumor activity of Saussurea Radix associated with inhibition of cell cycle progression and promotion of apoptosis caused by transcriptional regulation of p53, p21/Wafl and the other related genes was observed.

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Inhibition Effect of Chunglijagam-Tang on Invasion Activity of Human Lung Adenocarcinoma, A549 (청리자감탕(淸離滋坎湯)의 폐암 세포주 A549의 invasion activity 억제 효과)

  • Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Koo-Seok
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.7 no.1
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    • pp.109-116
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    • 2001
  • By applying in vitro invasion assay model, we examined the anti-metatstastic effect of ChunghjagamTang(CLJGT). In 3H-thymidine incorporation assay, CLJGT treated groups showed the decreased DNA synthesis rate compared with control group. Gelatin zymogram assay showed that CLJGT decreases the gelatinolytic activity of MMP-9 from A-549, at the concentration of $800{\mu}g/ml$. We examined whether CLJGT inhibits the invasion of A-549 cells through the matrigel precoated transwell chamber. The results showed that CLJGT effectively inhibited the invasion of A-549 as compared with the control (+PMA) groups. From our research, part of mechanism underlying anti-metastastic effect of CLJGT was proven in vitro.

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Circadian DNA-synthetic Rhythm Accompanied Mitotic Rhythm in Newly Hatched in Chicken Liver : Possible Role of Feeding Regiment (사료섭취가 병아리의 간세포증식 리듬에 미치는 영향)

  • Han, Joon-Pyo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.18 no.4
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    • pp.444-448
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    • 1989
  • The division and circadian DNA-synthesis rtythm are studied in chick liver immediately after hatching. The vision function is tested by the mitotic and the $^3H-thymidine$ labelling index The DNA synthesis exhibited a cyclic variation of 12 hours immediately after hatching. The rhythmic changes of DNA synthesis was maintained in the liver of meal-fed chickens, but the DNA synthetic activities decreased gradually in the starved chicken liver. From this time, the rhythm of DNA synthesis was greatly afffected by lighting schedule, all these DNA synthesis of chicken liver was accompained by mitosis.

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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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