• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.2
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• pp.300-308
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• 2000

The Fibroblast growth factors(FGFs) plays an important role in the control of osteogenesis during skeletal development. Especially, FGF-2 is a potent mesodermal inducer during embryogenesis and FGF receptors (FGFRs) messages are strongly expressed in developing bones. In this study, we investigated the effect of bFGF on osteopontin(OPN) gene expression in ST-2 cells and tried to elucidate the mechanism of its stimulatory effects. The obtain results were as follows; The treatment of bFGF(1ng/ml) upregulates OPN, fibronectin mRNA levels and downregulates type I collagen mRNA levels. But, there was no remarkable difference in alkaline phosphatase mRNA levels between two groups. The OPN gene expression increased in a dose-dependent manner up to 10ng/ml and OPN gene began to occur at around 3h with continuous increase up to 24h then decreased to basal level at 48h. 30 minutues pretreatment with cycloheximide (500ng/ml), a protein synthesis inhibitor, prior to addition bFGF resulted in blocking bFGF induced OPN expression. These results suggest that bFGF increased the level of OPN mRNA in a dose and time-dependent manner via the synthesis of certain transcriptional regulatory proteins.

• Journal of Ginseng Research
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• v.24 no.3
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• pp.134-137
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• 2000

Spontaneous bursting activity was studied in rat thalamocortical slices using extracellular field potential recording to test the potential utilization of ginsenoside Rb$_1$ in controlling overactivated neural systems. In order to induce bursting activity, slices were perfused with Mg$\^$2+/-free artificial cerebrospinal fluid (ACSF). Two major types of spontaneous bursting activity, simple thalamocortical burst complexes (sTBCs) and complex thalamocortical burst complexes (cTBCs), were recorded in Mg$\^$2+/ -free ACSF. Ginsenoside Rb$_1$ selectively suppressed cTBCs. Duration and occurrence rate of cTBCs were reduced by 87.3${\pm}$10.2% and 85.3${\pm}$ 14.7% in the presence of 90 ${\mu}$M ginsenoside Rb$_1$ respectively, while amplitude and intraburst frequency were slightly changed by ginsenoside Rb$_1$. In contrast, ginsenoside Rb$_1$was much less effective in reducing duration and occurrence rate of sTBCs. We also tested effects of ginsenoside Rb$_1$ on bursting activity in the presence of a GABA$\sub$A/ receptor antagonist, bicuculline methiodide (BMI). Ginsenoside Rb$_1$ had no effect in suppressing BMI-induced bursting activities. These results suggest that ginsenoside Rbi may be useful in controlling seizure-like bursting activity under pathological conditions.

• Parasites, Hosts and Diseases
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• v.29 no.2
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• pp.121-128
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• 1991

The degree of attraction of Toxoplasma gondii to vertebrate cells varies with cell type and cell phase. Human promyelocytic leukemia cells, HL-60, were synchronized by double thymidine block method and co-cultured with Toxoplasma for 1 hr at each cell stage to investigate the cell cycle specific susceptibility of parasites to host cells. For 30 hr the average number of Texoplasma that invaded was a little changed except at 3 hr from G1/S phase boundary which concurred with the peak point of DNA synthesis. At 3 hr which is a relatively short interval compared to whole S phase, modification of cells by parasitic invasion was most remarkable. The number of Toxoplasma that penetrated was increased to more than sin times. The shape of the cells became sludgy and almost indiscernible by strong accessibility of parasites only for an hour of mfd-S phase. The same auctuation was also observed at the second peak of S phase but weakly. This suggests that there be surface molecules concerning with the attachment of Texoplasma to the host cells, which is expressed at special point of S phase. further studies on the specific protein or similar molecules related could be carried out using synchronized HL-60 cells.

• Journal of Life Science
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• v.13 no.4
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• pp.529-534
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• 2003

This study was done to investigated the phytosterol compositions of safflower (Carthamus tinctorius L.) seed. The phytoestrogen activity was also determined using CAT-ELISA Kit in ethanol extract of safflower seed. The phytosterol of safflower seeds was identified using gas chromatography-mass spectrometry after saponification of the oils. The phytosterol content and composition of safflower seed oils were 4% and identified stigmast-5-en-3-ol (3$\beta$, 24S)-form, ${\gamma}$-sitosterol (clionasterol) with Wiley MS spectrum library. The synergistic effect of human estrogen receptor (hER) has been investigated using a minimal chimeric promoters composed of the TATA region of the adenovirus-2 major late promoter (A22MLP) and two consensus perfectly polindromic Xenopus vitellogenin A2 gene estrogen responsive elements (XVEREl19). Transient transfection experiments in tile human breast adenocarcinoma cell line MCF-7, which is known to express the estrogen receptor endogenously, revealed that phytoestrogen from Carthamus tinctorius L. acts as estrogen. We have observed the transcriptional activities stimulated methanol and ethanol extract of safflower seed in MCF-7, were 0.43 and 0.37 respectively, compared to that by $\beta$-estradiol as 1.0. Our data showed that safflower seeds have estrogenic activity methanol and ethanol extracts and ethanol lower than that of $\beta$-estradiol. This result provides the first evidence that the beneficial effect of safflower seeds may be mediated, at least in part, by the stimulating effect of phytoestrogen ell bone-protecting.

• Animal cells and systems
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• v.13 no.3
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• pp.275-281
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• 2009

The effects of pentylenetetrazol (PTZ), a GABA receptor antagonist, were studied on passive avoidance learning and expression of heat shock protein 70 (hsp70), neuroglobin, and fatty acid binding protein-7 (fabp-7) genes. Zebrafish were trained to stay in a dark compartment to avoid a weight dropping in an acryl shuttle box with a central sliding door. In two training sessions of 2 h interval, each consisting of 3 trials, the crossing time was significantly increased from $43.2{\pm}14.4s$ to $149.3{\pm}38.5s$ in the first training session and remained $116.1{\pm}36.0s$ s in the first trial of the second training session in the control. In zebrafish treated with PTZ before the first training session, the crossing time was significantly increased neither in the first nor in the second training session. However, the increased crossing time was maintained in the second training session when 10 mM PTZ was treated three times for 10 min at 30 min intervals between the first and second training session. Quantitative real-time PCR showed that expression level of hsp70 mRNA increased two to eight fold over that of control in the brain at 0-24 h after termination of PTZ treatment. No change in expression of neuroglobin and fabp-7 mRNA was shown in PTZ-treated zebrafish. Our studies suggest that PTZ impairs learning ability in avoidance response and also modifies expression of genes related to the neuroprotection.

• Molecules and Cells
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• v.24 no.1
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• pp.132-138
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• 2007

4-1BB (CD137), a member of the tumor necrosis factor receptor superfamily, is expressed on activated T-cells, and 4-1BB signaling due to interaction with 4-1BB ligand or ligation with anti-4-1BB monoclonal antibody (mAb) costimulates T cells. It has been shown that administration of anti-4-1BB mAb induces anti-tumor immunity in mice, but the nature of the cellular subsets responsible for this immunity is uncertain. In this study we found that anti-4-1BB mAb administration to B16F10 melanoma-bearing mice induced marked expansion of $CD11c^+CD8^+$ T-cells in parallel with suppression of pulmonary tumors. The mAb-treated mice produced higher levels of $IFN-{\gamma}$ in their tumor tissues, spleen and lymph nodes than mice exposed to control antibody. When the $CD11c^+CD8^+$ T-cells were purified and re-stimulated in vitro, they produced high levels of the Th1 cytokines, $IFN-{\gamma}$ and IL-2, but low levels of the Th2 cytokines, IL-4 and IL-10. Furthermore, they expressed high levels of 4-1BB and CD107a, a marker of activated cytotoxic T-lymphocytes. Our results suggest that $CD11c^+CD8^+$ T-cells play a role in the anti-tumor immunity induced by anti-4-1BB mAb.

• Development and Reproduction
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• v.22 no.4
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• pp.309-318
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• 2018

The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) activation by $17{\beta}$-estradiol ($E_2$) and progesterone ($P_4$) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL $E_2$, and $P_4$ with or without fetal bovine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and expression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of $E_2$ compared to 0.1% DMSO treatment in serum-free group (p<0.05). Then, $E_2$ and $P_4$ were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cultured cells without FBS. Only tPA mRNA was significantly increased by $E_2$ treatment (p<0.05). PAs activity was enhanced in $E_2$ treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to evaluate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.

• Biomolecules & Therapeutics
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• v.24 no.3
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• pp.291-297
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• 2016

Cytisine (CYT), a partial agonist of ${\alpha}4{\beta}2-nicotinic$ receptors, has been used for antidepressant efficacy in several tests. Nicotinic receptors have been shown to be closely associated with depression. However, little is known about the effects of CYT on the depression. In the present study, a mouse model of depression, the unpredictable chronic mild stress (UCMS), was used to evaluate the activities of CYT. UCMS caused significant depression-like behaviors, as shown by the decrease of total distances in open field test, and the prolonged duration of immobility in tail suspension test and forced swimming test. Treatment with CYT for two weeks notably relieved the depression-like behaviors in the UCMS mice. Next, proteins related to depressive disorder in the brain region of hippocampus and amygdala were analyzed to elucidate the underlying mechanisms of CYT. CYT significantly reversed the decreases of 5-HT1A, BDNF, and mTOR levels in the hippocampus and amygdala. These results imply that CYT may act as a potential anti-depressant in the animals under chronic stress.

• Journal of the East Asian Society of Dietary Life
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• v.26 no.3
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• pp.207-214
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• 2016

This study investigated the effects of ginger (Zingiber officinale Roscoe) on antioxidant and antiproliferative activities in A2058 human melanoma cells. The antioxidant and antiproliferative activities of 70% ethanol extracts of Zingiber officinale Roscoe were identified based on DPPH and ABTS free radical scavenging capacities. Treatment of cells with Zingiber officinale Roscoe at concentrations of 0, 0.2, and 0.4 mg/mL for 24 hours significantly reduced cell viability as determined by Hoechst 33258 nuclear staining, apoptosis analysis, and Western blotting analysis, respectively. In our study, 70% ethanol extracts of Zingiber officinale Roscoe exhibited antioxidant activity and inhibited A2058 cell growth in a dose-dependent manner. Concomitant activation of the mitochondria-dependent apoptotic pathway of A2058 human melanoma cells by Zingiber officinale Roscoe extracts was mediated via modulation of Bax and Bcl-2 expression, which activated cleavage of caspases-3, caspases-9, and poly ADP-ribose polymerase. The findings of study indicate that Zingiber officinale Roscoe extracts induce apoptosis in A2058 human melanoma cells, and this phenomenon occurs via the death receptor-mediated and intrinsic pathways.

• Clinical and Experimental Pediatrics
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• v.48 no.7
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• pp.779-788
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• 2005

Purpose : Echinacea, a traditional plant medicine has been used as immune-stimulant. Recent studies have revealed that extract of Echinacea has immunostimulatory effects on human blood mononuclear cells. This study was designed for the purpose of screening the genes associated with immunologic effects of Echinacea on monocytes and dendritic cells using a cDNA microarray chip. Methods : $CD14^+$ monocyte cells were cultured for one day with Echinacea extract(final concentration : $50{\mu}g/mL$) in experiment 1, but were cultured without Echinacea in experiment 2. The gene expression of these cultured monocytes was analyzed using the cDNA microarray chip. Dendritic cells produced from $CD14^+$ monocyte were cultured for five days with GM-CSF and IL-4, and then cultured for one day with Echinacea in experiment 3, but were done without Echinacea in experiment 4. Results : In experiments 1 and 2, there were 17 significantly expressed genes with average expression ratios above 2.5, including interferon gamma-inducible protein 30(IFI 30), CDC(cell-division-cylcle)-like kinase 2(CLK 2), syndecan binding protein(syntenin), superoxide dismutase 2, etc. In experiments 3 and 4, there were 24 gene, with significantly expressed genes were 24 genes, which were insulin-like growth factor 2(somatomedin A), methyl-CpG binding domain protein 3, IFI 30, small inducible cytokine subfamily A, member 22, etc. The genes encoding CD44, IFI 30, mannose receptor C type 1(MRC 1), chemokine receptor 7(CCR 7), CLK 2, syntenin and cytochrome C oxidase subunit VIII were significantly expressed in both monocytes and dendritic cells cultured with Echinacea. Conclusion : This study employed a cDNA microarray chip to elicit the immune-associated gene profile; the expression was enhanced by Echinacea in CD14+ monocytes and dendritic cells. Thus we laid the basis for the quantitative and functional analysis of genes induced by Echinacea in monocytes and monocyte-derived dendritic cells.