• YAKHAK HOEJI
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• v.46 no.1
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• pp.39-46
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• 2002

The water fraction exhibiting anticancer activity was prepared from 70% methanol extract of Artemisis argyi by stepwise solvent partioning. This water fraction(5 $\mu$g/ml concentration) showed a considerable cytotoxicity against leukemic L1210 cells with a maximal value of 92% for 3 days culture. Contrastingly to such substantial anticancer activities the identical fraction showed far low toxicity against normal lymphocytes than chloroform fraction of Artemisia argyi mitomycine and 5-fluorouracil at every concentration ranging 0.01$\mu$g/ml~10.00$\mu$g/ml. The cytotoxicity displayed against L1210 cells by the water fraction of Artemisia was found to be proportinal to the decrease of viability of L1210 cells. On the other hand, $O_2$ion generation in L1210 cells appeared to be elevated in accordance to cytotoxicity by the water fraction with concurrent increases of superoxide dismuatse (SOD) and glutathione peroxidase (GPx) which are responsible for the conversion of $O_2$ ion and $H_2O$$_2$ respectively These findings taken together indicate that the death of L1210 cells by the water fraction of Auemisia atgyi, may be induced at least in part by the detrimental action of reactive oxygen species (ROS) including $O_2$- in spite of substantial extorts of SOD and GPx to overcome the attack of ROS.

• Reproductive and Developmental Biology
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• v.29 no.2
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• pp.109-115
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• 2005

Hematopoietic stem cells (HSC) are multipotent cells that reside in the bone marrow and replenish all adult hematopoietic lineages throughoutthe lifetime. In this study, we analyzed the expression of receptors of $P2Y_{10}$, purinergic receptor families in murine hematopoietic stem cells, hematopoietic progenitor cells. In addition, the biological activity of $P2Y_{10}$ was investigated with B lymphocyte cell line, Ba/F3 in effect to cell growth and cell cycle. From the analysis of expression in hematopoieticstem cell. and progenitor with RT-PCR, $P2Y_{10}$ was strongly expressed in murine hematopoieticstem cells (c-kit+ Sca-l+ Lin-) and progenitor cell population, such as c-kit- Sca-l+ Lin-, c-kit+ Sca-l- Lin- and c-kit- Sca-l- Lin-. To investigate the biological effects by $P2Y_{10}$, retroviral vector from subcloned murine $P2Y_{10}$ cDNA was used fur gene introduction into Ba/F3 cells, and stable transfectant cells were obtained by flow cytometry sorting. In cell proliferation assay, the proliferation ability of $P2Y_{10}$ receptor gene­transfected cells was strongly inhibited, and the cell cycle was arrested at G1 phase. These result suggest that the $P2Y_{10}$ may be involved the biological activity in hematopoietic stem cells and immature B lymphocytes.

• Biomedical Science Letters
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• v.11 no.4
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• pp.487-492
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• 2005

The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. Spodoptera frugiperda Clone 21 (Sf2l) cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, very little is known about the properties of the endogenous sugar transporter(s) in Sf2l cells, although a saturable transport system for hexose uptake has been previously revealed in the Sf cells. In order to further examine the substrate and inhibitor recognition properties of the Sf2l cell transporter, the ability of hexoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. Transport was effectively inhibited by D-mannose and D-glucose. Of the hexoses tested, L-glucose had the least effect on 2dGlc transport in the Sf2l cells, indicating that the transport is stereoselective. Unlike the human HepG2 type glucose transport system, D-mannose had a somewhat greater affinity for the Sf2l cell transporter than D-glucose, implying that the hydroxyl group at the C-2 position is not necessary for strong binding. However, epimerization at the C-4 position of D-glucose (D-galactose) resulted in a dramatic decrease in affinity of the hexose for the Sf2l cell transporter. Such a lowering of affinity might be the result of the involvement of the C-4 hydroxyl in hydrogen bonding. It is therefore suggested that Sf2l cells were found to contain an endogenous sugar transport activity that in several aspects resembles the human HepG2 type glucose transporter, although the insect and human transporters do differ in their affinity for cytochalasin B.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.9
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• pp.1367-1374
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• 2005

Amino acid transporters play an important role in supplying nutrition to normal and cancer cells for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the $Na^+$-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-tyre amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In the present study, we have examined the expression and functional characterization of system L amino acid transporters in FOB human osteoblast cells. RT-PCR and western blot analysis have revealed that the FOB cells expressed LAT1, LAT2 together with their associating protein 4F2hc. The uptakes of $[^{14}C]_L$-leucine by FOB cells are $Na^+$-independent and almost completely inhibited by system L amino acid transporter selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). These results suggest that the transport of neutral amino acids including several essential amino acids for cellular nutrition into the FOB human osteoblast cells is mediated by system L amino acid transporters.

• Archives of Pharmacal Research
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• v.29 no.8
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• pp.645-650
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• 2006

Tributyltin chloride (TBTC) at concentrations of $0.5-1.0\;{\mu}M$ inhibits dopamine biosynthesis in PC12 cells. In this study, the effects of TBTC on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced cytotoxicity in PC12 cells were investigated. TBTC at concentrations up to $1.0\;{\mu}M$ neither affected cell viability, nor induced apoptosis after 24 or 48 h in PC12 cells. However, TBTC at concentrations higher than $2.0\;{\mu}M$ caused cytotoxicity through an apoptotic process. In addition, exposure of PC12 cells to non-cytotoxic (0.5 and $1.0\;{\mu}M$) or cytotoxic $(2.0\;{\mu}M)$ concentrations of TBTC in combination with L-DOPA (20, 50 and $100\;{\mu}M$) resulted in a significant increase in cell loss and the percentage of apoptotic cells after 24 or 48 h compared with TBTC or L-DOPA alone. The enhancing effects of TBTC on L-DOPA-induced cytotoxicity were concentration- and treatment time-dependent. These data demonstrate that TBTC enhances L-DOPA-induced cytotoxicity in PC 12 cells.

• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.545-555
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• 2002

Ginsenosides of 20(S)-protopanaxadiol and 20(S)-protopanaxatriol classification including the aglycones, PD, PI and ginsenosides Rh2, Rhl were shown to posses characteristic effects on proliferation of THP-l human leukaemia cells. A similar result was not apparent for ginsenoside Rg3 or dexamathasone. The concentration to inhibit $50\%$ of cells $(LC_{50})$ for PD, Rh2, PI and Rhl were 13 ${\mu}g/mL,\;15{\mu}g/mL,\;19{\mu}g/mL\;and\;210\;{\mu}g/mL$ respectively. Cell cycle analysis showed apoptosis with PD and PI treatment of THP-1 cells resulting in a build up of sub-G1 cells after 24, 48 and 72 hours of treatment. Rh2, and dexamathasone treatments also increased apoptotic cells after 24 hours, where as Rhl did not. After 48 and 72 hours Rh2, Rhl and dexamathasone similarly increased apoptosis, but these effects were significantly (P<0.05) lower than observed for both PD and PI treatments. Furthermore, treatments that produced the largest build up of apoptotic cells were also found to have the largest release of lactate dehydrogenase (LDH). It can be concluded from these studies that the presence of sugars to PD and PI aglycone structure reduces the potency to induce apoptosis, and alternately alter membrane integrity. These cytotoxic effects to THP-l cells were different from dexamethasone.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.6
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• pp.357-362
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• 2003

Carbon monoxide (CO) is low molecular weight oxide gas that is endogenously produced under physiological conditions and interacts with another gas, nitric oxide (NO), to act as a gastrointestinal messenger. The aim of this study was to determine the effects of exogenous CO on L-type calcium channel currents of human jejunal circular smooth muscle cells. Cells were voltage clamped with 10 mM barium ($Ba^{2+}$) as the charge carrier, and CO was directly applied into the bath to avoid perfusion induced effects on the recorded currents. 0.2% CO was increased barium current ($I_{Ba}$) by $15{\pm}2$% ($mean{\pm}S.E.$, p<0.01, n=11) in the cells. To determine if the effects of CO on barium current were mediated through the cGMP pathway, cells were pretreated with 1-H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, $10{mu}M$), a soluble guanylyl cyclase inhibitor, and exogenous CO (0.2%) had no effect on barium currents in the presence of ODQ ($2{\pm}1$% increase, n=6, p>0.05). CO mediates inhibitory neurotransmission through the nitric oxide pathway. Therefore, to determine if the effects of CO on L-calcium channels were also mediated through NO, cells were incubated with $N^G-nitro-L-arginine$ (L-NNA, 1 mM), a nitric oxide synthase inhibitor. After L-NNA pretreatment, 0.2 % CO did not increase barium current ($4{\pm}2$% increase, n=6, p>0.05). NO donor, SNAP ($20{\mu}M$) increased barium current by $13{\pm}2$% (n=6, p<0.05) in human jejunal smooth muscle cells. These data suggest that CO activates L-type calcium channels through NO/cGMP dependant mechanism.

• Journal of Practical Agriculture & Fisheries Research
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• v.14 no.1
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• pp.107-115
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• 2012

In juvenile size, the maximum clearance rate and ingestion rate were 15mm, which is the 70.8 mL/h, 4245.0×104cells/h, respectively. But the lowest clearance rate and ingestion rate were found at 5mm, which is the 4.1 mL/h, 246.0×104cells/h, respectively. The highest clearance rate and ingestion rate of AFDW seen at 7.5mm, with 3.3 mL/mg AFDW/h, 196.0×104cells/mg AFDW/h, respectively. According to the results of water temperature, clearance rate and ingestion rate were lowest at 10℃, showing the 0.3 mL/h, 20.3×104cells/h, respectively. The highest clearance rate and ingestion rate were 25℃, which is the 16.6 mL/h, 993.4×104cells/h, respectively. But in 30℃, clearance rate and ingestion rate decreased to the 12.9 mL/h, 772.6×104cells/h, respectively. With respect to the food density, the experiment groups with 30×104 cells/mL and 240×104 cells/mL showed the clearance rate of 21.1 mL/h and 2.3 mL/h, respectively. But the experimental group with 120×104 cells/mL or more showed decreased clearance rate. While, the maximum ingestion rate was at 60×104 cells/m, showing the 876.2×104cells/h.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.2
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• pp.311-315
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• 2002

Cellular death by apoptosis is an active process, depending on gene transcription and protein synthesis. It was reported that nitric oxide can induce apoptosis in several cancer cell-lines. We studied effects of Phytolacca esculentum van Houtt (Phytolaccaceae) Radix water extract (PRE) on the proliferation of transplanted-L1210 cells in mice. When PRE (500 mg/kg) was administered orally once a day for 7 days after transplantation of L1210 cells to mice, DNA fragmentation of transplanted-L1210 cells was induced and mitochondrial transmembrane potential of those cells was reduced. Additionally, DNA fragmentation of L1210 cells was induced by the treatment of PRE in vitro. Also, DNA fragmentation of L1210 cells was enhanced by co-culture with the peritoneal macrophages obtained from PRE-administered mice and was partly inhibited by L-NMMA in vitro. PRE enhanced the production of nitric oxide and tumor necrosis factor-α from peritoneal rnacrophages. These results suggest that PRE induces apoptosis of transplanted-L1210 cells via directive action on L1210 cells and stimulation of nitric oxide and tumor neaosis factor-α from macrophages.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.2
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• pp.235-240
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• 2000

Inhibitory effects of cockle extracts on carcinogen-induced cytotoxicity in C3H/10T1/2 cells were studied. Soup (62$\mu\textrm{g}$/mL), solubility (28$\mu\textrm{g}$/mL) and liposolubility (9 $\mu\textrm{g}$/mL) of the cockle inhibited 3-methyl-cholanthrene(MCA)-induced cytotoxicity in C3H/10T1/2 cells by 53 and 94%, respectively. These results suggest that the extracts cockle might have anticarcinogen-induced cytotoxicity of C3H/10T1/2 cells. The effects of cockle extracts on the immune response related to its antitumor activity in vitro and in vivo were investigated. The cockle extracts showed a direct cytotoxic effect on sarcoma-180 cells, tumor cells in vitro. Soup (0.49 mg/mL), solubility (0.11 mg/mL) and liposolubiliy (0.05 mg/mL) of the cockle markedly decreased the total numbers of sarcoma-180 cells, but not their viability. The phagocytic acitivity of peritoneal macrophage of mice was significantly augmented by these extracts of the cockle compared with that of control in vivo. These extracts also raised the phagocuytic index, indicating that the number of phagocytize dmicrobes per macrophage increased. Thus, cockle extracts might show a antitumor activity by enhancing the phagocytic cell activities.