• Title/Summary/Keyword: five dose assay

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Development of a Noble Dosimetry Using Metaphase Analysis and Micronuclei Assay of Bone Marrow Cells in Mice (마우스 골수세포의 중기염색체 분석 및 미소핵 검사를 이용한 피폭선량 평가법의 개발)

  • Min, Jung-Jun;Bom, Hee-Seung;Kim, Young-Ho;Yoon, Hyun-Joong;Kim, Ji-Yeul
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.1
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    • pp.74-81
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    • 2000
  • Purpose: The purpose of this study was to develop in vivo dosimetries using both chromosomal aberrations and micronuclei in mice to assess biological effects of radiations. Materials and Methods: Five each mice were irradiated with 0, 1, 2, 3, 4, 5, 10 Gy of Cs-137 gamma-rays. We scored numbers of chromosomal aberrations in metaphase spreads and numbers of micronuclei in bone marrow smears under light microscope, and obtained the dose-response relationships. We also examined the relationship between the two dose-response curves. Results: The frequency of both chromosomal aberrations and micronuclei increased with dose, in a linear-quadratic manner The delta, beta, and alpha coefficients were 0.0176, 0.0324, and 0.0567 for metaphase analysis (r=1.0, p<0.001) and 0.0019, 0.0073, and 0.0506 for micronuclei assay (r=1.0, p<0.001). The frequency of chromosomal aberrations and micronuclei in different radiation doses was significantly correlated (r=0.99, p<0.01). Conclusion: In vivo dosimetry using either metaphase analysis or micronucleus assay was feasible in mice. These methods could be useful to evaluate biological effects of radiation.

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Anti-allergic and Anti-inflammatory Effects of Jacho(Lithospermum Erythrorhizon) (자초가 항알러지 염증반응에 미치는 영향)

  • Kwon Mi-Hwa;Lee Jin-Yong;Kim Deog-Gon
    • The Journal of Korean Medicine
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    • v.26 no.3 s.63
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    • pp.110-123
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    • 2005
  • Objectives : This study was performed to examine the anti-allergic and anti-inflammatory effects of Jacho (Lithospermum erythrorhizon). Methods : Macrophage 264.7 cells were pretreatment, macrophage were incubated with lipopolysaccharide(LPS) 100ng/ml for 12h ($TNF-{\alpha}$, IL-6) or 24h ($IL-1\beta$, IL-10) and media collectred and $TNF-{\alpha}$, IL-6, $IL-1{\beta}$, and IL-10 concentrations in supernatants were each measured by enzyme-linked immunosorbent assay. Concentrations of Jacho used were 50, 100, 250, 500, and $1000{\mu}g/ml$, and hydrocortisones used were 10-8, 10-7, 10-6, 10-5, and 10-4M. Results : Jacho showed inhibitory effect on $TNF-{\alpha}$ LPS-stimulated macrophage 264.7. The inhibitory effect was most significant in $250{\mu}g/ml$, and was not in a dose-dependent manner as in the hydrocortisone group Jacho also showed inhibitory effect on IL-6 by LPS-stimulated macrophage 264.7. The inhibitory effect was most significant in $1000{\mu}g/mL$, and increased in a roughly dose-dependent manner. Jacho and hydrocortisone showed contrary effect on $IL-1\beta$. Jacho obviously increased the expression of $IL-1\beta$, in alt five concentrations, End at the fewest concentration $(50{\mu}g/ml)$ the level of $IL-1\beta$, was highest. On the other hand, hydrocortisone was observed to have inhibitory effect on $IL-1\beta$, in all five concentrations. IL-10 was obviously inhibited by Jacho and hydrocortisone respectively in a roughly dose-dependent manner. Conclusions : By the findings of this experiment. Jacho was observed to have anti-allergic and anti-inflammatory effects through inhibiting pro-inflammatory cytokine $TNF-{\alpha}$ and IL-6, and might be one of the effective therapeutic regimens for allergic diseases.

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Cytotoxicity of Some Natural Products in Human Cancer Cells (수종의 한국산 야생식물에서의 항암효과 검색)

  • Park, Sung-Hee;Cho, Sung-Ji;Rhee, In-Sun;Lee, Chong-Ock;Kim, Chong-Won;Rhee, In-Ja
    • Korean Journal of Pharmacognosy
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    • v.27 no.4
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    • pp.383-388
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    • 1996
  • The objective of this reseach is to find new antitumoral substances from natural products. Several of natural products have been used as food that were isolated into hexane(Hex.) and/or ethylacetate(EtOAc) extracts. we have tested cytotoxicities of these plants against human solid tumor cells. The cytotoxic activity of these plants were tested using Sulforhodamin B(SRB) assay. Hexane extracts of Chrysanthemum sinense, Allium tubersum. Beta vulgaris, Ixeris dentata have revealed cytotoxicities against five human solid tumor cells, and its cytotoxicities of each cell line were $10-100\;{\mu}l/ml$ ED50 (Effective dose that cause 50% inhibition of cell growth in vitro)

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Antioxidative Effect of Proteolytic Hydrolysates from Ecklonia cava on Radical Scavenging Using ESR and $H_2O_2$-induced DNA Damage

  • Heo, Soo-Jin;Park, Pyo-Jam;Park, Eun-Ju;Cho, So-Mi K.;Kim, Se-Kwon;Jeon, You-Jin
    • Food Science and Biotechnology
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    • v.14 no.5
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    • pp.614-620
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    • 2005
  • The antioxidative effect of Ecklonia cava, a brown marine alga, was investigated on radical scavenging, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), and hydroxyl and alkyl radicals, using an electron spin resonance (ESR) technique, and on the inhibition of $H_2O_2$-induced DNA damage using comet assay. E. cava was enzymatically hydrolyzed with five food industrial proteases (Alcalase, Flavourzyme, Kojizyme, Neutrase and Protamex) to prepare water-soluble extracts. All the proteolytic hydrolysates exhibited strong dose-dependent radical scavenging activities (above 80%) at a concentration of $2.5\;{\mu}g/mL$. Kojizyme extract (obtained by proteolytic hydrolysation of E. cava with Kojizyme) showed the highest hydroxyl radical scavenging activity of around 98%. In addition, the $H_2O_2$-induced DNA damage was determined using a comet assay, which was quantified by measuring the tail length. Reduction of DNA damage increased with increasing concentrations of Kojizyme extract from E. cava. These results indicated that E. cava has a potential as a valuable natural antioxidative source.

Effects of Sophorae Radix on Human Colorectal Adenocarcinoma Cells (고삼의 인체 대장암세포에 미치는 효과)

  • Kim, Min-Chul;Lee, Hee-Jung;Lim, Bo-Ra;Kim, Hyung-Woo;Kim, Byung-Joo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.2
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    • pp.155-159
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    • 2012
  • The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of 5-Fluorouracil (5-FU) in human colorectal adenocarcinoma cells (HT-29). We used human colorectal adenocarcinoma cell line, HT-29 cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle (sub G1) analysis was done the HT-29 cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with $150{\mu}g$ Sophorae Radix. Sophorae Radix inhibited the growth of HT-29 cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in HT-29 cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with 5-FU markedly inhibited the growth of HT-29 cells compared to Sophorae Radix or 5-FU alone. After 3 days treatment of HT-29 cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Our findings provide insight into unraveling the effects of Sophorae Radix in human colorectal adenocarcinoma cells and developing therapeutic agents against colorectal cancer.

Effects of Apoptosis of Sophorae Radix on Human Gastric Adenocarcinoma cells (인체 위암세포에서 고삼의 세포사멸효과)

  • Lim, Bo-Ra;Lee, Hee-Jung;Kim, Min-Chul;Kim, Hyung-Woo;Kim, Byung-Joo
    • Korean Journal of Oriental Medicine
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    • v.18 no.1
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    • pp.85-92
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    • 2012
  • Objective : The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of 5-Fluorouracil (5-FU) in human gastric adenocarcinoma cells (AGS). Method : We used human gastric adenocarcinoma cell line, AGS cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle (sub G1) analysis was done the AGS cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with 100 ${\mu}g/ml$ Sophorae Radix. Result : Sophorae Radix inhibited the growth of AGS cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in AGS cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with 5-FU markedly inhibited the growth of AGS cells compared to Sophorae Radix or 5-FU alone. After 3 days treatment of AGS cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Conclusion : Our findings provide insight into unraveling the effects of Sophorae Radix in human gastric adenocarcinoma cells and developing therapeutic agents against gastric cancer.

Effects of Sophorae Radix on Human Breast Adenocarcinoma Cells (고삼의 인체 유방암세포에 미치는 효과)

  • Lee, Hee-Jung;Kim, Min-Chul;Lim, Bo-Ra;Bae, Go-Eun;Kim, Hyung-Woo;Kwon, Young-Kyu;Kim, Byung-Joo
    • Korean Journal of Oriental Medicine
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    • v.18 no.1
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    • pp.75-84
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    • 2012
  • Objective : The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of Doxorubicin (DOX) in human breast adenocarcinoma cells (MCF-7). Method : We used human breast adenocarcinoma cell line, MCF-7 cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle analysis was done the MCF-7 cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with 100 ${\mu}g/ml$ Sophorae Radix. Result : Sophorae Radix inhibited the growth of MCF-7 cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in MCF-7 cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with DOX markedly inhibited the growth of MCF-7 cells compared to Sophorae Radix or DOX alone. After 3 days treatment of MCF-7 cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Conclusion : Our findings provide insight into unraveling the effects of Sophorae Radix in human breast adenocarcinoma cells and developing therapeutic agents against breast cancer.

In Vitro Antitumor Properties of an Isolate from Leaves of Cassia alata L

  • Olarte, Elizabeth Iglesias;Herrera, Annabelle Aliga;Villasenor, Irene Manese;Jacinto, Sonia Donaldo
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3191-3196
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    • 2013
  • Leaf extracts of Cassia alata L (akapulko), traditionally used for treatment of a variety of diseases, were evaluated for their potential antitumor properties in vitro. MTT assays were used to examine the cytotoxic effects of crude extracts on five human cancer cell lines, namely MCF-7, derived from a breast carcinoma, SK-BR-3, another breast carcinoma, T24 a bladder carcinoma, Col 2, a colorectal carcinoma, and A549, a nonsmall cell lung adenocarcinoma. Hexane extracts showed remarkable cytotoxicity against MCF-7, T24, and Col 2 in a dose-dependent manner. This observation was confirmed by morphological investigation using light microscopy. Further bioassay-directed fractionation of the cytotoxic extract led to the isolation of a TLC-pure isolate labeled as f6l. Isolate f6l was further evaluated using MTT assay and morphological and biochemical investigations, which likewise showed selectivity to MCF-7, T24, and Col 2 cells with $IC_{50}$ values of 16, 17, and 17 ${\mu}g/ml$, respectively. Isolate f6l, however, showed no cytotoxicity towards the non-cancer Chinese hamster ovarian cell line (CHO-AA8). Cytochemical investigation using DAPI staining and biochemical investigation using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-a method used to detect DNA fragmentation-together with caspase assay, demonstrated apoptotic cell death. Spectral characterization of isolate f6l revealed that it contained polyunsaturated fatty acid esters. Considering the cytotoxicity profile and its mode of action, f6l might represent a new promising compound with potential for development as an anticancer drug with low or no toxicity to non-cancer cells used in this study.

Effect of Kp, an Antitumor Protein-Polysaccharide from Mycelial Culture of Phellinus Linteus on the Humoral lammune Response of Tumor-Bearing ICR Mice to Sheep Red Blood Cells

  • Chung, Kyeong-Soo;Kim, Shin-Sook;Kim, Hee-Soo;Kim, Kye-Yang;Han, Man-Woo;Kim, Kil-Hyun
    • Archives of Pharmacal Research
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    • v.16 no.4
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    • pp.336-338
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    • 1993
  • The immunomodulating activity of Kp, an antitumor protein-polysacchanide preparation from the shake-cultured mycelia of Phellinus linteus, was investigated in ICR mice subcutaneously implanted wit $1\times10^6$ cells of sarcoma 180. The mice were intraperitoneally administered with Kp at a does of 100 mg/kg once daily for five consecutive days starting from 24 hrs after the tumor implantation. Ten days after the last injection, the mice were immunized with $1\times10^7$ or $4\times10^8$ sheep red blood cells (SRBC) and five days later, the antibody-forming immune response were assessed by direct hemolytic plaque assay. To an immunization does of $1\times10^7$ SRBC, the Kp-treated mice elicied a successful humoral immune response despite the turmor-burden and produced $259\times10^3$ plaque-forming cells (PFC)/spleen, while the corresponding tumor-bearing control mice showed virtually no reponse $(2.0\times10^3$ PFC/spleen) (the stimulation index=129.5). However, to an immunization dose of $4\times10^8$ SRBC, both of the control mice and Kp-treated mice showed almost the same level of strong humoral immune response. From these data it is clear that Kp effectively restores the humoral immune response of the turmor-bearing ICR mice.

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Bio-antimutagenic effects of water extract from Rehmannia glutinosa Liboschitz in SOS Chromotest (SOS Chromotest에서 숙지황 물 추출물의 세포내 항돌연변이 효과)

  • Ahn, Byung-Yong;Lee, Kap-Sang;Maeng, Il-Kyung;Song, Geun-Seoub;Choi, Dong-Seong
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.439-445
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    • 1998
  • The antimutagenic activity of the water extract of Rehmannia glutinosa Liboschitz (RG) on the mutagenicity induced by 4-nitroquinoline 1-oxide (4-NQO), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), mitomycin C (MMC), $aflatoxin\;B_1\;(AFB_1)$ and benzo(a)pyrene [B(a)P] were studied using the SOS Chromotest with Escherichia coli PQ37. The water extract of RG was separated into methanol soluble and methanol insoluble parts. The methanol soluble part exhibited higher inhibition effects than the methanol insoluble part against the mutagenic activities of five mutagens. Step-wise fractionation of methanol soluble part was done using methanol, ethyl acetate and water. Among these fractions, water fraction had the strongest inhibitory effects against the mutagenenicity of five model mutagens, showing $4.5{\sim}29.5%$ inhibition, but the $AFB_1$ mutagenic potency was increased slightly by ethyl acetate fraction. The water fraction was further partitioned by sephadex LH-20 column chromtography, and 9 subfractions were obtained. The fraction III showed the strongest inhibitory effects with dose response against the mutagenic activities induced by all the tested chemical mutagens. The inhibition rates of fraction III at concentration of $400\;{\mu}g/assay$ were 29%, 35%, 38%, 25% and 24% against 4-NQO, MNNG, MMC, AFBl and B(a)P, respectively. The fraction III also exhibited a strong bio-an-timutagenicity against 4-NQO and $AFB_1$ by showing more than 40% inhibition.

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