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Cell Viability and Flow Cytometry Analysis of a Novel Antitumor Agent, Heptaplatin in Human Melanoma Cell Line, SK-MEL-28 (신규항암제인 Heptaplatin의 인체 흑색종세포(SK-MEL-28)에 대한 세포생존률 및 유세포 분석)

  • 최수라;명평근
    • YAKHAK HOEJI
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    • v.47 no.6
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    • pp.345-351
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    • 2003
  • Heptaplatin, cis-Malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1,3-dioxolane]platinum(II), is a novel platinum-based antitumor agent with clinical potential against human stomach cancer and the 3rd generation of the cisplatin. This study was performed to study how cisplain, heptaplatin and sunpla which is a mixture of heptaplatin and mannitol (w: w=l : 2) affect cell viability of SK-MEL-28 human melanoma cell line. Heptaplatin ($IC_{50}$/; 95.35 $\mu$M) and sunpla ($IC_{50}$/; 10.95 11M) were less effect than cisplatin (IC $_{50}$; 10.92 $\mu$M) on the SK-MEL-28 cells. By cell cycle analysis using flow cytometry, it was identified that the cells were arrested at G2/M phase by cisplatin, heptaplatin and sunpla, and percentage of cell death group was increased according to increasing of time and concentration. These results suggest that cisplatin, heptaplatin and sunpla are a novel anticancer agent against human melanoma cell.l.

Effects of TAM (Taraxacum mongolicum) on Th2 Cytokine Production in MC/9 Mast Cells (포공영(蒲公英)이 MC/9 mast cell에서의 Th2 cytokine 발현에 미치는 영향)

  • Jang, Moon-Hee;Choi, Jae-Song;Bae, Na-Young;Ahn, Teak-Won
    • Journal of Sasang Constitutional Medicine
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    • v.24 no.1
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    • pp.54-65
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    • 2012
  • 1. Objective : The purpose of this study is to investigate the effects of TAM (Taraxacum mongolicum) on Th2 cytokine production in MC/9 mast cells. 2. Methods : The effects of TAM was analyzed by ELISA and Real-time PCR in MC/9 mast cells. Levels of IL-5, IL-13 were measured using enzyme-linked immunosorbent assays(ELISA). mRNA levels of IL-4, IL-5, IL-6, IL-13 were analyzed with Real-time PCR. 3. Results : 1) TAM inhibited the IL-4 production significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$, $200{\mu}g/ml$. 2) TAM inhibited the IL-13 production significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$, $200{\mu}g/ml$. 3) TAM inhibited the IL-4 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 4) TAM inhibited the IL-5 mRNA expression significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$. 5) TAM inhibited the IL-6 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 6) TAM inhibited the IL-13 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 4. Conclusions : These results indicate that TAM (Taraxacum mongolicum) has the effect of decreasing the Th2 cytokine production in the MC/9 mast cell.

Antitumor Activity of Arylacetylshikonin Analogues

  • Kim, Seon-Hee;Song, Gyu-Yong;Jin, Guang-Zhu;Ahn, Byung-Zun
    • Archives of Pharmacal Research
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    • v.19 no.5
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    • pp.416-422
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    • 1996
  • Twenty one phenylacetylshikonin analogues were synthesized from various subsitituted phenyl acetic acids and their cytotoxicity values against A549, K562 and L1210 cell lines and antitumor action in mice bearing S-180 cells were measured. All of phenylacetylshikonin analogues expressed a potent cytotoxicity $(ED_{50}, 0.1-1.80{\mu}g/ml)$ against L1210 and K562 cells. L1210 cells were the most sensitive to shikonin analogues among these cells. Except 4-methosyphenylacetylshikonin $(0.098 {\mu}g/ml)$, and a-acetoxyphenylacetylshikonin $(0.10 {\mu}g/ml)$, all other shikonin derivatives sshowed higher $ED_{50}$ values than phenylacetylshikonin $(0.13{\mu}g/ml)$, in L1210. In K562 cell, a-substitution of phenylacetylshikonin $(0.1{\mu}g/ml)$, while other subsitutions increased it slightly; 4-methoxyphenylacetylshikonin $(0.033{\mu}g/ml)$ showed a exceptionally good cytotoxicity against K562 cell. 4-Halogenation tended to decrease the cytotoxic effect on L1210 cells, while it enhanced the effect on K562; 4-bromophenylacetyl $$[ED_{50};(L1210)=1.76{\mu}g/ml, ;ED_{50};(K 562)=0.32 {\mu}g/ml]$$ and 4-chlorophenylacetyl shikonin $$[ED_{50};(L1210)=1.64 {\mu}g/ml, ;ED_{50};(K562)=0.32 {\mu}g/ml]$$. In contrast, A549 cells were much less sensitive to these shikonin analogues which showed $ED_{50}$ values of$1.5-1.35 {\mu}g/ml)$.Most of phenylacetylshikonin derivatives showed good antitumor activity in mice bearing S-180 cells. a-A-cetoxyphenylacetylshikonin and 4-dimethylaminophenylacetylshikonin showed highest T/C value (192-195%), implying that introduction of a-acetyl or of 4-dimethylamino group enhanced the antitumor activity as shown for 4-dimethylaminophenylacetylshikonin (T/C, 192%). It might be due to improvement of water solubility by dimethylamino group in the molecule.

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Cytotoxicity and DNA Damage Induced by Magnetic Nanoparticle Silica in L5178Y Cell

  • Kang, Jin-Seok;Yum, Young-Na;Park, Sue-Nie
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.261-266
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    • 2011
  • As recent reports suggest that nanoparticles may penetrate into cell membrane and effect DNA condition, it is necessary to assay possible cytotoxic and genotoxic risk. Three different sizes of magnetic nanoparticle silica (MNP@$SiO_2$) (50, 100 and 200 nm diameter) were tested for cytotoxicity and DNA damage using L5178Y cell. MNP@$SiO_2$ had constant physicochemical characteristics confirmed by transmission electron microscope, electron spin resonance spectrometer and inductively coupled plasma-atomic emission spectrometer for 48 h. Treatment of MNP@$SiO_2$ induced dose and time dependent cytotoxicity. At 6 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$ compared to vehicle control (p<0.05 or p<0.01). Moreover, at 24 h, 50 or 100 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$(p<0.01). And treatment of 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Furthermore, at 48 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Cellular location detected by confocal microscope represented they were existed in cytoplasm, mainly around cell membrane at 2 h after treatment of MNP@$SiO_2$. Treatment of 50 nm MNP@$SiO_2$ significantly increased DNA damage at middle and high dose (p<0.01), and treatment of 100 nm or 200 nm significantly increased DNA damage in all dose compared to control (p<0.01). Taken together, treatment of MNP@$SiO_2$ induced cytotoxicity and enhanced DNA damage in L5178Y cell.

Effects of enamel matrix protein derivatives on the periodontal ligament like fibroblast and osteoblast like cells (법랑기질 단백질 유도체가 치주인대양세포 및 조골양세포에 미치는 영향)

  • Kim, Dong-Woon;Chung, Chin-Hyung;Lim, Sung-Bin;Ko, Seon-Yle
    • Journal of Periodontal and Implant Science
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    • v.33 no.2
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    • pp.225-246
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    • 2003
  • Recent study on the enamel matrix derivatives explained on the effects of new bone and new attachment formation in infrabony pocket of periodontal defects. The purpose of this study was to investigate on the biological effects of enamel matrix derivatives to attachment, proliferation and activation of periodontal ligament and osteoblast cells, After treatment of osteoblast and PDL cells with various Emdogain concentration level(0.03${\mu}g$/ml, 3${\mu}g$/ml, 300${\mu}g$/ml), activation of osteogenetic factor, calcified nodule formation and measuring alkaline phosphatase activity(ALP) were performed. 1. Both osteoblast and PDL cell showed increasing initial cell attachment with 300${\mu}g$/ml Emdogain concentration. 2. At the level of 300${\mu}g$/ml, accelerated proliferation of oseoblast and PDL cell was appeared. 3. As Emdogain's concentration increased, increased ALP activation of osteoblast was shown. In case of PDL cell, Emdogain increased ALP activation prominently at the level of 300${\mu}g$/ml. 4. No statistically significant activating change were founded at all of the concentrations of Emdogain on the activating of transcript factor Runx2 for differentiating osteoblast. 5. At the level of 300${\mu}g$/ml, calcified nodule formation was increased prominently to compare with other concentration. These results indicated that Emdogain should activate initial attachment, proliferation and activation, but not on Runx2 activation and can be used for useful tool of the treatment of periodontal tissue regeneration.

Regulation of Inflammatory Cytokine Production by Bee Venom in Rat Chondrocytes

  • Kim, Eun-Jung;Kim, Gye-Yeop
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.1
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    • pp.132-137
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    • 2011
  • Bee venom acupuncture (BVA), as a kind of herbal acupuncture, involved injecting diluted bee venom into acupoints and is used for pain, osteoarthritis and rheumatoid arthritis patients. BVA is growing in popularity, especially in Korea, and is used primarily for pain relief in many kinds of diseases. However, the effect of bee venom anti-inflammatory related action in lipopolysaccharide (LPS) induced chondrocyte stress have not been reported yet. The aim of this study was to investigate the effect of bee venom of cell viability and inflammatory cytokine in rat articular chondrocyte cultures stimulated with lipopolysaccharide. Inflammation was induced in rat chondrocytes by treatment with $10{\mu}g/m{\ell}$ LPS. The change of cell viability were decreased in chondrocytes after treatment with lipopolysaccharide. The cell viability revealed that BV exerted no significant cytotoxicity in the rat chondrocyte. Bee venom inhibited decreased cell viability in the presence of lipopolysaccharide ($10{\mu}g/m{\ell}$) in a dose dependent manner(0.1, 0.5, 1.0 and $5.0{\mu}g/m{\ell}$) at bee venom(p<0.05). Tumor necrosis factor (TNF)-${\alpha}$ production in the presence of lipopolysaccharide($1{\mu}g/m{\ell}$) was also inhibited in a dose dependent manner (p<0.05 from bee venom $0.1{\mu}g/m{\ell}$). Interleukin (IL)-6 production in the presence of lipopolysaccharide ($10{\mu}g/m{\ell}$) was inhibited as well (p<0.05 at bee venom 0.1, 0.5, 1.0 and $5.0{\mu}g/m{\ell}$, respectively). Our results demonstrate that bee venom was a anti-inflammatory agent of chondrocytes. Bee venom may exert its anti inflammatory effects through inhibition of TNF-${\alpha}$ and IL-6 synthesis, and may then pain relief and reduce the articular destruction.

The Comparative Study on Anti-inflammatory Antioxidant and Antibacterial Effects with Regard to the Extraction Solvents of Ligustici Rhizoma (고본(藁本)의 추출용매에 따른 항염, 항산화 및 항균 효과에 대한 비교 연구)

  • HwangBo, Min;Seo, Hyung-Sik
    • Journal of Pharmacopuncture
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    • v.14 no.1
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    • pp.71-78
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    • 2011
  • Objective : This study was performed to compare anti-inflammation, anti-oxidation and anti-bacterial effects of Ligustici Rhizoma (LR) extracted with two kinds of solvents, ethanol and distilled water. Methods : It is prepared two kinds of LR extracts 20, 50, 100 ${\mu}l/mg$ by first. MTT assay way to measure cytotoxicity is formed in Raw 264.7 cell. The anti-inflammation effect is measured by ability to inhibit production of NO in Raw 264.7 cell. The anti-oxidation effect was measured by DPPH Radical scavenging ability in HaCaT cell. The anti-bacterial effect was measured by inhibition zone diameter on Propionibacterium acnes. Results : 1. LR (20 ${\mu}l/mg$) extracted with ethanol was showed 80% cytotoxicity, LR (50 ${\mu}l/mg$) extracted with ethanol and LR (20, 50 ${\mu}l/mg$) extracted with water were showed 70% cytotoxicity, LR (100 ${\mu}l/mg$) extracted with ethanol and LR (100 ${\mu}l/mg$) extracted with water were showed 60% cytotoxicity in Raw 264.7 cell. 2. LR (100 ${\mu}l/mg$) extracted with ethanol was showed more significantly inhibitory effect on NO production than the water extraction. 3. Two kinds of LR extraction groups did not show significantly scavenging effect of DPPH radicals. 4. Two kinds of LR extractions did not have a inhibitory effect on Propionibactrium acnes. Conclusion : Two kinds of LR extracts have not cytotoxicity, statistically significant ability to scavenge DPPH radicals and effect to inhibit Propionibactrium acnes. LR extracted with ethanol only have a little effect to inhibit NO production. This study proposes that LR extracted with ethanol is more effective in anti-inflammation.

NF-${\kappa}B$ Inhibitor Suppresses Hypoxia-induced Apoptosis of Mouse Pancreatic ${\beta}$-cell Line MIN6

  • Koh, Hyun Sook;Kim, Jae Young
    • Biomedical Science Letters
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    • v.20 no.1
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    • pp.14-24
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    • 2014
  • Hypoxia is one of the main reasons for islet apoptosis after transplantation as well as during isolation. In this study, we attempted to determine the potential usefulness of NF-${\kappa}B$ inhibitor for suppression of hypoxia-induced ${\beta}$-cell apoptosis as well as the relationship between IP-10 induction and ${\beta}$-cell apoptosis in hypoxia. To accomplish this, we cultured the mouse pancreatic ${\beta}$-cell line MIN6 in hypoxia (1% $O_2$). Among several examined chemokines, only IP-10 mRNA expression was induced under hypoxia, and this induced IP-10 expression was due to NF-${\kappa}B$ activity. Since a previous study suggested that IP-10 mediates ${\beta}$-cell apoptosis, we measured hypoxia-induced IP-10 protein and examined the effect of anti-IP-10 neutralizing Ab on hypoxia-induced ${\beta}$-cell apoptosis. However, IP-10 protein was not detected, and anti-IP-10 neutralizing Ab did not rescue hypoxia-induced MIN6 apoptosis, indicating that there is no relationship between hypoxia-induced IP-10 mRNA expression and hypoxia-induced ${\beta}$-cell apoptosis. Since it was still not clear if NF-${\kappa}B$ functions as an apoptotic or anti-apoptotic mediator in hypoxia-induced ${\beta}$-cell apoptosis, we examined possible involvement of NF-${\kappa}B$ in hypoxia-induced ${\beta}$-cell apoptosis. Treatment with 1 ${\mu}M$ NF-${\kappa}B$ inhibitor suppressed hypoxiainduced apoptosis by more than 50%, while 10 ${\mu}M$ AP-1 or 4 ${\mu}M$ NF-AT inhibitor did not, indicating involvement of NF-${\kappa}B$ in hypoxia-induced ${\beta}$-cell apoptosis. Overall, these results suggest that IP-10 is not involved in hypoxia-induced ${\beta}$-cell apoptosis, and that NF-${\kappa}B$ inhibitor can be useful for ameliorating hypoxia-induced ${\beta}$-cell apoptosis.

Study on the analysis of disproportionate data and hypothesis testing (불균형 자료 분석과 가설 검정에 관한 연구)

  • 장석환;송규문;김장한
    • The Korean Journal of Applied Statistics
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    • v.5 no.2
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    • pp.243-254
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    • 1992
  • In the present study two sets of unbalanced two-way cross-classification data with and without empty cell(s) were used to evaluate empirically the various sums of squares in the analysis of variance table. Searle(1977) and Searle et.al.(1981) developed a method of computing R($\alpha$\mid$\mu, \beta$) and R($\beta$\mid$\mu, \alpha$) by the use of partitioned matrix of X'X for the model of no interaction, interchanging the columns of X in order of $\alpha, \mu, \beta$ and accordingly the elements in b. An alternative way of computing R($\alpha$\mid$\mu, \beta$), R($\beta$\mid$\mu, \alpha$) and R($\gamma$\mid$\mu, \alpha, \beta$) without interchanging the columns of X has been found by means of,$(X'X)^-$ derived, using $W_2 = Z_2Z_2-Z_2Z_1(Z_1Z_1)^-Z_1Z_2$. It is true that $R(\alpha$\mid$\mu,\beta,\gamma)\Sigma = SSA_W and R(\beta$\mid$\mu,\alpha,\gamma)\Sigma = SSB_W$ where $SSA_W$ and means analysis and $R(\gamma$\mid$\mu,\alpha,\beta) = R(\gamma$\mid$\mu,\alpha,\beta)\Sigma$ for the data without empty cell, but not for the data with empty cell(s). It is also noticed that for the datd with empty cells under W - restrictions $R(\alpha$\mid$\mu,\beta,\gamma)_W = R(\mu,\alpha,\beta,\gamma)_W - R(\mu,\alpha,\beta,\gamma)_W = R(\alpha$\mid$\mu) and R(\beta$\mid$\mu,\alpha,\gamma)_W = R(\mu,\alpha,\beta,\gamma)_W - R(\mu,\alpha,\beta,\gamma)_W = R(\beta$\mid$\mu) but R(\gamma$\mid$\mu,\alpha,\beta)_W = R(\mu,\alpha,\beta,\gamma)_W - R(\mu,\alpha,\beta,\gamma)_W \neq R(\gamma$\mid$\mu,\alpha,\beta)$. The hypotheses $H_o : K' b = 0$ commonly tested were examined in the relation with the corresponding sums of squares for $R(\alpha$\mid$\mu), R(\beta$\mid$\mu), R(\alpha$\mid$\mu,\beta), R(\beta$\mid$\mu,\alpha), R(\alpha$\mid$\mu,\beta,\gamma), R(\beta$\mid$\mu,\alpha,\gamma), and R(\gamma$\mid$\mu,\alpha,\beta)$ under the restrictions.

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BCSC(Buired contact Solar cel1)의 제조를 위한 laser scribing Laser scrining for Buired contact Solar ell

  • 조은철;지일환;이수홍
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1995.05a
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    • pp.154-159
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    • 1995
  • To achieve a high aspect ration of metal contact, buried contact solar cell scribe the silicon surface using laser. The Q-switched NdLYAG laser which has 1.064$\mu\textrm{m}$ wavelength use for silicon scribing with 25~40$\mu\textrm{m}$ width and 20~200$\mu\textrm{m}$ depth capabilities. The 2~3% shading losses are very low campared to the screen printing solar cell. In this paper, we investigate the silicon scribing theory and pratice, scribing system for BCSC processing.

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