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Anti-Inflammatory Effect of Ligustri Lucidi Fructus Water Extract in RAW 264.7 Cells Induced by LPS (여정실(女貞實)이 LPS로 유발된 RAW 264.7 cell의 염증에 미치는 영향)

  • Lee, Yong-Hyun;Lim, Eun-Mee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.4
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    • pp.66-81
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    • 2013
  • Purpose: This study was carried out to investigate the anti-inflammatory effects of Ligustri Lucidi Fructus water extract (LF) in the lipopolysaccharide (LPS)-induced mouse macrophages RAW 264.7 cell. Methods: Ligustri Lucidi Fructus was extracted with distilled water (2,000 ml) for 2 hours. In order to evaluate cytotoxicity of LF, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. To investigate anti-inflammatory effects of LF, the concentration of nitric oxide (NO) was measured with NO assay, cytokine was measured by Bio-Plex cytokine assay, and intracellular calcium (Ca) was measured with Fluo-4 Ca assay in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically (P<0.05). Results: 1. LF showed no cytotoxicity. 2. LF inhibited significantly the production of NO at the concentration of 25, 50 and $100{\mu}g/ml$. 3. LF inhibited significantly the production of interleukin (IL)-4, macrophage inflammatory protein (MIP)-$1{\alpha}$, granulocyte colony stimulating factor (G-CSF) at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 4. LF inhibited significantly the production of granulocyte macrophage-colony stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF) at the concentration of 50, 100 and $200{\mu}g/ml$, the interferon (IFN)-${\gamma}$ at 25, 50 and $100{\mu}g/ml$ respectively. 5. LF inhibited significantly the production of IL-$1{\beta}$ at the concentration of 50 and $200{\mu}g/ml$, the IL-5 at 25 and $100{\mu}g/ml$, the IL-12p70, MIP-$1{\beta}$ at 50 and $100{\mu}g/ml$, the regulated on activation, normal T cell expressed and secrete d (RANTES) at 100 and $200{\mu}g/ml$ respectively. 6. LF inhibited significantly the production of IL-10, interferon gamma-induced protein (IP)-10 at the concentration of $200{\mu}g/ml$. 7. LF inhibited significantly the production of intracellular Ca at the concentration of 25, 50, 100 and $200{\mu}g/ml$. Conclusions: These results suggest that LF has anti-inflammatory effect and immuno-modulating activity.

Antioxidant Activity of Green Tea Extracts toward Human Low Density Lipoprotein (사람의 Low Density Lipoprotein에 대한 녹차의 항산화 활성)

  • Park, Chun-Ok;Jin, Seung-Heun;Ryu, Beung-Ho
    • Korean Journal of Food Science and Technology
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    • v.28 no.5
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    • pp.850-858
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    • 1996
  • Green tea leaves 12.5 g were extracted twice with 500 ml boiling water. The green tea extract (GTE) contained 4.67 mg solid. The GTE contained polyphenols sush as 54.12% (-) epicatechin gallate, 26.21% (-) epicatechin, 10.71% epicatechin gallate, 7.09% (-) epicatechin and 1.85% catechin. The GTE inhibited the copper-catalyzed oxidation of human LDL at the concentrations of 50 and $100\;{\mu}g/ml$ GTE in the presence of $5\;{\mu}M$ $CuSO_{4}$. The electrophoretic mobility of the LDL oxidized in the presence of $5\;{\mu}M\;CuSO_{4}$ was higher than that of the native LDL. The GTE also inhibited LDL oxidation induced by J774, human monocyte-derived macrophages and vascular endotherial cells. The LDL modified by copper or cells was inhibited by human macrophages at a much greater rate than native LDL in the presence of GTE. The GTE was found to be a potent inhibitor of modification of LDL. GTE inhibited the uptake of cell-modified $^(125)I-labelled$ LDL by macrophages. The formation of conjugated dienes was strongly inhibited in the presence of 50 or $100\;{\mu}g/ml$ GTE.

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Biomass of Primary Producer in the Ch$\check{o}$nsu Bay -Relationships between Phytoplankton Carbon, Cell Number and chlorophyll- (천수만 일차생산자의 생물량 -식물플랑크톤 탄소량과 세포개체수 및 클로로필과의 관계-)

  • Shim, Jae Hyung;Shin, Yoon Keun
    • 한국해양학회지
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    • v.24 no.4
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    • pp.194-205
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    • 1989
  • In order to study the biomass of primary producer, phytoplankton is collected monthly September 1985 to August 1986 in Ch$\check{o}$nsu Bay. Phytoplankton carbon contents which are calculated from phytoplankton volume were ranged from $26.7{\mu}gC/l$ to $960.7{\mu}gC/l$, and average carbon contents of each month lie in the range of $58.6-684.7{\mu}gC/l$(annual mean $208.5{\mu}gC/l$). For net plankton analysis with the carbon contents, cell numbers, and chlorophyll concentrations show a close correlation, while for nanoplankton the correlation was low, indicating that nano-fraction includes a significant portion of picoplankton. Also, the multiple regression analysis with carbon content, cell number, and chlorophyll concentration to size fraction well illustrate the prime importance of the net-fraction in phytoplankton group. C/Chl-a ratios ranged from 9.1 to 100.5, average rations of net- and nanoplankton are 111 and 6.4, respectively. The greater net plankton faction is, the higher C/Chl-a ratio is, however in case of high nanoplankton portion C/Chl-a ratio show low level. These results indicate that the difference of C/Chl-a ratio per phytoplankton cell size be main factor for the variation of C/Chl-a ratio in Ch$\check{o}$nsu Bay. As C/Chl-a ratio fluctuates greatly in coastal ecosystem, that use of a direct conversion of convert chlorophyll to organic carbon may lead erronous estimation.

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Cytotoxic Effect of Radioprotective Ginseng Protein Fraction on CHO-KI Cells (방사선 방어작용이 있는 인삼 단백분획의 CHO-KI 세포에 대한 세포 독성)

  • Kim, Choon-Mi;Yoon, Suk-Ran
    • YAKHAK HOEJI
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    • v.32 no.5
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    • pp.313-318
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    • 1988
  • Radioprotective ginseng protein fraction was isolated from Korean white ginseng and its cytotoxic effect on CHO-K1 cells was studied by the method of measuring the relative cell survival and total cellular protein content (FRAME method). When ginseng protein at the dose of 300, 600, 900, $1200{\mu}g/ml$ was treated to cells for 24 hrs, the relative survival was significantly decreased at the concentration of above $600{\mu}g/ml$, indicating the presence of cytotoxic effect of the protein at certain concentration. When cellular protein content was measured after ginseng protein at the dose of 300, 600, 900, $1200\;{\mu}g/ml$ was treated, the amount of cellular protein was significantly reduced at the concentration above $600{\mu}g/ml$ in the case of 24 hr treatment and at all concentrations including $300{\mu}g/ml$ in the case of 72 hr treatment. The data suggest that the protein may inhibit cell growth, resulting in the reduction of live cells in culture. $ID_{50}$ value which is the concentration of ginseng protein that reduces the total cellular protein content to 50% of the control was calculated as 2276.86 and $1323.32\;{\mu}g/ml$ in groups treated for 24 and 72 hr, respectively. Since $ID_{50}$ value of above $1000{\mu}g/ml$ indicates very weak cytotoxicity, the ginseng protein seems to exert very weak cytotoxic effect on CHO-K1 cells.

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The Role of Nitric Oxide on the Growth Regulation of Chinese Cabbage (Brassica campestris L.) Primary Leaves (배추 (Brassica campestris L.) 제 1엽의 생장조절에 대한 Nitric Oxide의 역할)

  • Ham Jeong-Hun;Jin Chang-Duck
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.293-300
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    • 2005
  • The possible role of nitric oxide (NO)-induced cell division was investigated to explain the physiologycal effects of a NO donor, sodium nitroprusside (SNP) on the growth of primary leaves in chinese cabbage seedling plants. Exogenous treatment of SNP to chinese cabbage plants for 8 days at different concentrations (0, 200, 500 and 1000 ${\mu}M$) affected the leaf growth in a concentration-dependent manner, showing a maximum growth at $200\;{\mu}M$. In accordance with leaf growth responses, the chlorophyll and soluble protein contents increased strongly to 142% and 134% of control at $200\;{\mu}M$ SNP, respectively. However, a very little decrease in chlorophyll and a 13%> decrease in protein were observed at $1000\;{\mu}M$ SNP. In addition, the content of DNA and RNA also increased maximumly to 142% and 139% of the control at $200\;{\mu}M$ SNP, respectively, whereas they decreased to 80% and 84% of the control at $1000\;{\mu}M$ SNP. With respect to the development of enzymes related to cell wall synthesis, $200\;{\mu}M$ SNP led to the maximum activities in both phenylalanine ammonia-lyase (212% of the control) and guaiacol peroxidase (134% of the control). However, the activities of both enzymes were not modified significantly at $1000\;{\mu}M$ SNP. In conclusion, these results suggest that the enhancement of leaf growth in chinese cabbage plants by SNP at the effective concentration was probably due to the NO ability in the induction of cell division.

Effects of Donor Somatic Cell Conditions on In Vitro Development of Nuclear Transplanted Porcine Embryos (돼지 공여세포의 조건이 핵이식 수정란의 체외발달에 미치는 영향)

  • 홍승표;박준규;이명열;이지삼;정장용
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.213-221
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    • 2001
  • This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.

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Effective Utilization of Hemp Fiber for Pulp and Papermaking (I) -Morphological Characteristics of Hemp Fiber- (펄프.제지용 원료로서의 삼 섬유 이용에 관한 연구(제 1보) -대마 구성 세포의 현미경적 관찰-)

  • Yoon, Seung-Lak;Lee, Myoung-Ku
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.42 no.1
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    • pp.7-12
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    • 2010
  • Morphological characteristics of hemp fiber were investigated using a light microscope in order to provide fundamental data for the use of hemp as a papermaking law material. Phloem of hemp is composed of cortical parenchyma cells and bast fiber with thick walls while xylem is composed of vessel, wood fiber and ray parenchyma cells. Also there are solitary pore and radial pore multiple which exist in diffuse porous pattern. Ray cells consist of uniseriate rays and thin walled ray parenchyma cells. Wood fibers are composed of three types: a large diameter fiber with longer length; a large diameter fiber with shorter length; a small diameter fiber with medium length. Vessel elements are composed of: a medium length one; a longer length one; the one whose both end walls have ligules or tails. Parenchyma cells in xylem and pit parenchyma cells have completely different size and shape. For bast fiber, the average length is about 4.4 mm and the width is about $30.5\;{\mu}m$; for vessel element, $600.0\;{\mu}m$ in length and $493.6\;{\mu}m$ in width; for wood fiber, $1000\;{\mu}m$ and $38.9\;{\mu}m$; for parenchyma cell, $50\;{\mu}m$ and $26.4\;{\mu}m$.

Toxicological Effects of B(a)P on Preimplantation Mouse Embryos in Vitro (in vitro에서 B(a)P이 착상전 마우스 배자에 미치는 독성학적 영향에 관한 연구)

  • 박귀례;이유미;김판기;신재호;강태석;김주일;장성재
    • Journal of Environmental Health Sciences
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    • v.24 no.2
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    • pp.126-133
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    • 1998
  • Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

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A Study of Neuroproctective Effect of Bupleuri Radix on Hippocampal Neurons (시호(柴胡)의 뇌해마 신경세포 보호효능에 대한 연구)

  • Lee, Won-Chul;Shin, Kwang-Sik
    • The Journal of Internal Korean Medicine
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    • v.25 no.4
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    • pp.227-241
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    • 2004
  • Objective : This study was performed to investigate neuroprotective effects of Bupleuri Radix against oxidative and ischemic damages. Method : To observe the neuroprotective effects against ischemic damage, ischemic insult was induced by oxygen/glucose deprivation (OGD) on organotypic hippocampal slice cultures (OHSC) from 1 week-old Sprague-Dawley rats. Propidium iodide (PI) fluorescence-stained neuronal dead-cell areas, area percentages and TUNEL-positive apoptotic cells in CA1 and dentate gyrus, and LDH levels in culture media of the OHSC were measured following Bupleuri Radix extract treatment. Result : The following results were obtained: (1) The $5\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in CA1 region of the OHSC from 18 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ of Bupleuri Radix treatment was also significant from 6 hrs to 48 hrs following the OGD and was more effective. (2) The 5 and $50\;{\mu}g/ml$ of Bupleuri Radix treatment demonstrated a significant decrease in PI fluorescence-stained neuronal dead-cell areas and area percentage in DG region of the OHSC from 6 hrs to 48 hrs following the OGD. The $50\;{\mu}g/ml$ treatment was more effective than the $5\;{\mu}g/ml$ treatment. (3) Bupleuri Radix treatment demonstrated a significant decrease in TUNEL-positive apoptotic cells in CA1 region (with 5 and $50\;{\mu}g/ml$) and in DG region (with $50\;{\mu}g/ml$) of the OHSC damaged by the OGD. (4) Bupleuri Radix treatment demonstrated a significant decrease in LDH concentrations in culture media of the OHSC damaged by the OGD. Conclusion : These results suggest that Bupleuri Radix has neuroprotective and control effects on inflammatory and immune responses where there has been ischemic damage to the central nervous system.

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Evaluation of Cytotoxicity, Carbohydrate, and Lipid Inhibitory Activity of Codonopsis lanceolata using Different Solvent Fractions

  • Boo, Hee-Ock;Park, Jeong-Hun;Kim, Seung-Mi;Woo, Sun-Hee;Park, Hyeon-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.4
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    • pp.498-503
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    • 2015
  • This study was conducted to evaluate the cytotoxicity and ${\alpha}-Amylase$, ${\alpha}-Glucosidase$, pancreatic lipase inhibition in vitro by different solvent fractions from the roots of Codonopsis lanceolata. The values of $IC_{50}$ against Calu-6 cell showed a high effect in n-hexane fraction ($10.13{\mu}g/mL$) whereas DW fraction exhibited the weakest inhibition on cell viability, having an $IC_{50}$ value of over $1,000{\mu}g/mL$. The values of $IC_{50}$ against HCT-116 cell showed the highest activity in n-BuOH fraction ($102.01{\mu}g/mL$), followed by n-hexane fraction ($145.85{\mu}g/mL$), methylene chloride fraction ($332.02{\mu}g/mL$), ethyl acetate fraction ($462.93{\mu}g/mL$) and DW fracion ($>1,000{\mu}g/mL$). ${\alpha}-Amylase$ inhibitory activity in methylene chloride fraction and ethyl acetate fraction was found to have a higher inhibitory effect with 24.5% and 25.6% than the other fractions. The highest ${\alpha}-Glucosidase$ inhibitory activity was observed from the ethyl acetate fraction extract, while the extract of DW fraction showed the lowest level of inhibitory activity at given experiment concentration. The pancreatic lipase inhibitory activity of C. lanceolata was found to have a higher the effect in ethyl acetate fraction. Inhibition of lipase activity of the ethyl acetate fraction and n-hexane fraction showed a relatively high, while the extract of DW fraction showed the lowest level at given experiment concentration. These results suggested that the roots of C. lanceolata may assist in the potential biological activity on carbohydrate, lipid Inhibitory activity and anticancer activity.