• Title/Summary/Keyword: C3H mouse

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The Production Mechanism of TNF-${\alpha}$ and IL-6 by Group IIA Phospholipase $A_2$ (IIA형 Phospholipase $A_2$에 의한 TNF-${\alpha}$와 IL-6 생성 기전)

  • Ahn, Jae-Hong;Park, Dae-Won;Kim, Jin-Hee;Bae, Jei-Jun;Bae, Yeun-Kyoung;Park, Yoon-Ki
    • Journal of Yeungnam Medical Science
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    • v.21 no.2
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    • pp.177-190
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    • 2004
  • Background: Secretory phospholipase $A_2$ ($sPLA_2$) are a group of extracellular enzymes that release fatty acids at the sn-2 position of phospholipids. Group IIA $sPLA_2$ ($sPLA_2$-IIA) has been detected in the inflammatory fluids, and its plasma level increases in the inflammatory disease. This study examined the effect of $sPLA_2$-IIA on mouse macropahges in order to investigate the potential mechanism of $sPLA_2$-induced inflammation. Materials and Methods: Wild type $PLA_2$ and mutant H48Q $PLA_2$ were purified from HEK293 cells transfected with the corresponding plasmids, and the $PLA_2$ activities were measured using 1-palmitoyl-2-[1-$^{14}C$]linoleoyl-3-phosphatidylethanolamine as substrates. The TNF-${\alpha}$ and IL-6 released in the supernatants were determined by ELISA. In addition, the TNF-${\alpha}$ and IL-6 mRNA were analyzed by RT-PCR. Results: $sPLA_2$-IIA stimulated the production of TNF-${\alpha}$ and IL-6 in a dose- and time-dependent manner. In addition, the effect of $sPLA_2$-IIA on cytokine production from the macrophage was found to be associated with the accumulation of their specific mRNA. The mRNA levels of TNF-${\alpha}$ and IL-6 peaked at 2 and 6 hours in a time-dependent manner, respectively. Conclusion: In conclusion, the production of proinflammatory cytokine might be mediated by the binding of $sPLA_2$-IIA to the receptors.

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Validation of a HPLC MS/MS Method for Determination of Doxorubicin in Mouse Serum and its Small Tissues (마우스 혈장과 조직에서의 doxorubicin 측정 HPLC-MS/MS 방법)

  • Park, Jung-Sun;Kim, Hye-Kyung;Lee, Hye-Won;Lee, Mi-Hyun;Kim, Hyun-Gi;Chae, Soo-Wan;Chae, Han-Jung
    • Korean Journal of Clinical Pharmacy
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    • v.16 no.1
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    • pp.23-27
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    • 2006
  • Doxorubicin (DXR) is a type of anti-cancer drug called an 'anthracycline glycoside', It works by impairing DNA synthesis, a crucial feature of cell division, and thus is able to target rapidly dividing cells. Doxorubicin is a very serious anti-cancer medication with definite potential to do great harm as well as great good. A liquid chromatography-tandem mass spectroscopy (LC-MS/MS) method was developed to identify and quantify DXR in small-volume biological samples. After the addition of internal standard (IS, $5{\mu}L\;of\;1{\mu}M/ml$ daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for a 1min and centrifugation at 15,000g for 10 min the organic phase was transferred and evaporated under a vacuum. The residue was reconstituted with $350{\mu}L$ of mobile phase and $10{\mu}L$ was injected into C18 column with mobile phase composed of 0.05M ammonium acetate (0.1 M acetic acid adjusted to pH 3.5) and acetonitrile (40:60, v/v). The flow rate was kept constant at $350{\mu}L/min$. The ions were quantified in the multiple reaction mode (MRM), using positive ions, on a triple quadrupole mass spectrometer. The lower limits of quantification for Doxorubicin in plasma and small tissues were approximately 0.5 ng/mL and 0.5 ng/mL respectively. Intra- and inter-assay accuracy (% of nominal concentration) and precision (% CV) for all analytes were within 15%, respectively.

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Biological Markers as Predictors of Radiosensitivity in Syngeneic Murine Tumors (동계 마우스 종양의 방사선 감수성 예측인자로서의 생물학적 표지자)

  • Chang Sei-Kyung;Kim Sung-Hee;Shin Hyun-Soo;Seong Jin-Sil
    • Radiation Oncology Journal
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    • v.24 no.2
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    • pp.128-137
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    • 2006
  • Purpose: We investigated whether a relationship exists between tumor control dose 50 ($TCD_{50}$) or tumor growth delay (TGD) and radiation induced apoptosis (RIA) in syngeneic murine tumors. Also we investigated the biological markers that can predict radiosensitivity in murine tumor system through analysis of relationship between $TCD_{50}$, TGD, RIA and constitutive expression levels of the genetic products regulating RIA. Materials and Methods: Syngeneic murine tumors such as ovarian adenocarcinoma, mammary carcinoma, squamous cell carcinoma, fibrosarcoma, hepatocarcinoma were used In this study. C3H/HeJ mice were bred and maintained in our specific pathogen free mouse colony and were $8{\sim}12$ weeks old when used for the experiments. The tumors, growing in the right hind legs of mice, were analyzed for $TCD_{50}$, TGD, and RIA at 8 mm in diameter. The tumors were also analyzed for the constitutive expression levels of $p53,\;p21^{WAF1/CIP1},\;BAX,\;Bcl-2,\;Bcl-X_L,\;Bcl-X_S$, and p34. Correlation analysis was peformed whether the level of RIA were correlated with $TCD_{50}$ or TGD, and the constitutive expression levels of genetic products regulating RIA were correlated with $TCD_{50}$, TGD, RIA. Results: The level of RIA showed a significant positive correlation (R=0.922, p=0.026) with TGD, and showed a trend to correlation (R=-0.848), marginally significant correlation with $TCD_{50}$ (p=0.070). It indicates that tumors that respond to radiation with high percentage of apoptosis were more radiosensitive. The constitutive expression levels of $p21^{WAF1/CIP1}$ and 34 showed a significant correlation either with $TCD_{50}$ (R=0.893, p=0.041 and R=0.904, p=0.035) or with TGD (R=-0.922, p=0.026 and R=-0.890 p=0.043). The tumors with high constitutive expression levels of $p21^{WAF1/CIP1}$ or p34 were less radiosensitive than those with low expression. Conclusion: Radiosensitivity may be predicted with the level of RIA in murine tumors. The constitutive expression levels of $p21^{WAF1/CIP1}$ or p34 can be used as biological markers which predict the radiosensitivity.

The Effect of Ginkgo Biloba Extract on Radiosensitivity of Mouse Skin and Jejunal Crypt (Ginkgo Biloba Extract가 마우스 피부 및 공장 소낭선의 방사선감수성에 미치는 영향)

  • Shin, Kyung-Hwan;Ha, Sung-Whan
    • Radiation Oncology Journal
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    • v.16 no.2
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    • pp.107-114
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    • 1998
  • Purpose : Ginkgo biloba extract(GBE) is known to increase the peripheral blood circulation. This study was designed to evaluate the effect of GBE on the acute normal tissue radiation reaction. Materials and Methods : mice were divided into two groups, radiation alone and two doses GBE plus radiation, for both acute skin reaction and jejunal crypt assay. GBE was given i.p. one hour before irradiation with priming dose given one day earlier. Thirty to Fifty Gy for acute skin reaction and 11 to 14 Gy for jejunal crypt were irradiated to right hind leg and whole body, respectively. Results : Radiation doses($RD_{50}$) for Peak skin score of 2.0 were 44.2Gy (40.6-48.2Gy) for radiation alone and 44.4Gy(41.6-47.4Gy) for two doses GBE plus radiation, showing no effect of GBE on acute radiation skin damage. The numbers of regenerating jejunal crypts per circumference were also almost the same for each radiation dose level(p=0.57-0.94), and the mean lethal doses($D_o$) were 1.800y(1.57-2.09Gy) for radiation alone and 1.88Gy(1.65-2.18Gy) for two doses GBE plus radiation, indicating no effect of GBE on jejunal crypt cell survival after radiation. Conclusion : GBE doesn't increase acute normal tissue radiation reaction in this model system. As GBE was verified to enhance radiation effect on tumor, high therapeutic gain is expected when GBE is combined with radiation therapy.

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Anti-wrinkle Activity of a Curdrania tricuspidata Extract on Ultraviolet-induced Photoaging (Hairless Mice를 이용한 광노화 모델에서 꾸지뽕나무 추출물의 피부주름 형성 억제효과)

  • Kim, Ok Kyung;Ho, Jin-Nyoung;Nam, Da-Eun;Jun, Woojin;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.4
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    • pp.608-614
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    • 2013
  • We investigated the anti-wrinkle activity of an 80% ethanol extract of Curdrania tricuspidata leaves (CTL80) on ultraviolet-induced photoaging in hairless mice. Skin wrinkles were induced by 10 weeks of UVB-irradiation on the back of Skh-1 hairless mice three times a week. Mice were divided into ten groups; normal control (-UVB), UVB irradiated control group (+UVB), dietary groups (UVB+ascorbic acid 0.1%, UVB+CTL80 0.1%, UVB+CTL80 0.25%) and topical application groups (-UVB+base lotion (BL), UVB+BL, UVB+ascorbic acid 1%+BL, UVB+CTL80 1%+BL, UVB+CTL80 2%+BL). Wrinkle formation, histological changes, superoxide dismutase (SOD) activities, glutathione peroxidase (GSH-Px), and the expression of matrix metalloproteinases (MMP-1, MMP-3 and MMP-9) were analyzed. Wrinkles for the +UVB groups formed as a pattern of deep furrows and thick crests. Wrinkles with CTL80 treatment formed as a pattern of shallow furrows and thin crests, with wrinkle areas were lower than the +UVB group. In an antioxidant analysis of mouse blood, SOD and GSH-Px activities were significantly higher in the CTL80 topical application group compared to the +UVB group. The mRNA expression of MMPs in the +UVB group was significantly higher than the normal control group, and significantly lower in the CTL80-treated group. In conclusion, CTL80 exerted anti-wrinkle activity on ultraviolet-induced photoaging by regulating antioxidative defense systems and MMPs expression.

Antioxidant Activity and Protective Effects of Cirsium japonicum against Damaged Mouse Liver Cell (BNL CL.2) (엉겅퀴의 항산화 활성 및 손상된 흰쥐 간세포(BNL CL.2)에 대한 간 보호 효과)

  • Kim, Seonjeong;Kang, Seungmi;Ko, Keonhee;Nam, Sanghae
    • Journal of Life Science
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    • v.27 no.4
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    • pp.442-449
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    • 2017
  • We analyzed the content of total phenolic and silymarin compounds of Cirsium japonicum (CJ), and its antioxidant activities and Liver protective effects were compared with those of Silybum marianum (SM). The total phenolic content in the aerial part ($97.22{\pm}5.51mg/g$) of CJ is higher than that in the underground part ($85.32{\pm}3.06mg/g$). The total silymarin content of CJ was 55.56% of SM, with the underground part ($0.47{\pm}0.03mg/g$) having higher content than the aerial part ($0.18{\pm}0.02mg/g$). The antioxidant activity of CJ was generally slightly lower than that of milk thistle, and the underground part of CJ generally had higher activity compared to the aerial part. When CJ extracts were processed at 1 mg/ml, DPPH activities were $83.76{\pm}0.60$ and $88.28{\pm}0.17%$, and FRAP activities were $77.63{\pm}0.70$ and $82.83{\pm}0.39%$ for extracts from aerial part and underground part, respectively. ABTS activities were $68.60{\pm}1.24$ and $63.41{\pm}0.57%$ for underground and aerial part respectively when extracts were processed at 0.1 mg/ml. The Liver protective effects of CJ were higher in the extracts from underground part compared to the aerial part, Liver cells were damaged by treating them with t-BHP, $H_2O_2$ and Ethanol, and then they were treated with 0.2 mg/ml CJ extracts. The survival rates of the damaged liver cells were $49.58{\pm}0.34$, $76.87{\pm}1.10$ and $71.73{\pm}0.58%$ respectively, which were higher than the cells not treated with extracts.

Natural killer cell activity in mice infected with Acanthamoeba culbertsoni (Acanthamoeba culbertsoni 감염에 있어 자연살세포의 활성)

  • Hyeon, Dong-Geun;Sin, Ju-Ok;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.101-112
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    • 1992
  • The natural killer cell activity of splenocytes and TBC, active NK cells, recycling capacity of natural killer cells were observed by means of both the 51Cr-release cytotoxicity assay and single cell cytotoxicity assay against YAC-1. CSH/HeJ mice were infected intranasally with $1{\times}10^4{\;}or{\;}1{\times}10^5$ trophosoites of pathogenic Acanthamoeba culbertsoni. The infected mice showed mortality rate of 34% in $1{\times}10^4$ group and 65% in 1{\times}10^5 group, and mean survival time was $16.40{\pm}3.50$ {\;}and{\;}3.20{\pm}4.09$ days respectively. The cytotoxic activity of natural killer cells of the 2 groups was significantly higher than that of non-infected mice from the 12th hour to the 2nd day after infection, showing the highest on the first day. On the l0th day after infection, the cytotoxic activity of natural killer cells was significantly suppressed as compared with that of the control. There was no significant difference in NK cell cytotoxicity between two infected groups. The targetbinding capacity and active NK cells of natural killer cells in $1{\times}10^5$ trophosoite infected mice was significantly increased on the 12th hour and the first day after infection as compared with the control group. Maximal recycling capacity (MRC) was not changed during the observation period. The present results indicated that the elevation of natural killer cell activity in the mice infected with A. culbertsoni was due to elevation of target.binding capacity and increased active NK cells of natural killer cells, and not due to the maximal recycling capacity of the individual NK cell, and there was no difference between two experimental dose groups.

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Radioprotective Effects of Granulocyte-Colony Stimulating Factor in the Jejunal Mucosa of Mouse (생쥐에서 과립구 집락형성인자(Granulocyte-Colony Stimulating Factor)의 공장점막에 대한 방사선 보호효과)

  • Ryu, Mi-Ryeong;Chung, Su-Mi;Kay, Chul-Seung;Kim, Yeon-Shil;Yoon, Sei-Chul
    • Radiation Oncology Journal
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    • v.19 no.1
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    • pp.45-52
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    • 2001
  • Purpose : Granulocyle-colony stimulating factor (G-CSF) has been widely used to treat neutropenia caused by chemotherapy or radiotherapy. The efficacy of recombinant human hematopoietic growth factors in improving oral mucositis after chemotherapy or radiotherapy has been recently demonstrated in some clinical studies. This study was designed to determine whether G-CSF can modify the radiation injury of the intestinal mucosa in mice. Materials and Methods : One hundred and five BALB/c mice weighing 20 grams were divided into nine subgroups including G-CSF alone group $(I:10\;{\mu}g/kg\;or\;II:100\;{\mu}g/kg)$, radiation alone group (7.5 or 12 Gy on the whole body), combination group with G-CSF and radiation (G-CSF I or II plus 7.5 Gy, G-CSF I or II plus 12 Gy), and control group. Radiation was administered with a 6 MV linear accelerator (Mevatron Siemens) with a dose rate of 3 Gy/min on day 0. G-CSF was injected subcutaneously for 3 days, once a day, from day -2 to day 0. Each group was sacrificed on the day 1, day 3, and day 7. The mucosal changes of jejunum were evaluated microscopically by crypt count per circumference, villi length, and histologic damage grading. Results : In both G-CSF I and II groups, crypt counts, villi length, and histologic damage scores were not significantly different from those of the control one (p>0.05). The 7.5 Gy and 12 Gy radiation alone groups showed significantly lower crypt counts and higher histologic damage scores compared with those of control one (p<0.05). The groups exposed to 7.5 Gy radiation plus G-CSF I or II showed significantly higher crypt counts and lower histologic damage scores on the day 3, and lower histologic damage scores on the day 7 compared with those of the 7.5 Gy radiation alone one (p<0.05). The 12 Gy radiation plus G-CSF I or II group did not show significant difference in crypt counts and histologic damage scores compared with those of the 12 Gy radiation alone one (p>0,05). Most of the mice in 12 Gy radiation with or without G-CSF group showed intestinal death within 5 days. Conclusion : These results suggest that G-CSF may protect the jejunal mucosa from the acute radiation damage following within the tolerable ranges of whole body irradiation in mice.

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