• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.203-207
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• 2004

The transgenic mice carrying human Interleukin-10 (hIL-10) gene in conjunction with bovine (3 -casein promoter express hIL-10 in milk during lactation. In this study, stability of germ line transmission and expression of hIL-10 transgene integrated into host chromosome were monitored up to generation F8 of transgenic mice. When male mouse of generation F8 was crossbred with normal females, approximately half of offspring (50.9±5.8%) were identified as transgenic mice. Generation F9 to F15 mice also showed similar transmission rates (66.0±20.1%, 61.5±16.7%, 41.1±8.4%, 40.7±20.3%, 61.3±10.8%, 49.2±18.8% and 43.8±25.9%, respectively), implying that hIL-10 transgene can be transmitted stably up to long term generation in the transgenic mice. Expression levels of human IL-10 from milk of generation F9 to F14 mice were 3.6± 1.2 mg/ml, 4.2±0.9 mg/ml, 5.7±1.5 mg/ml, 6.3±3.5 mg/ml, 6.8±4.5 mg/ml and 6.8±3.1 mg/ml, respectively, which was showed high-level expression compared with that of generation F1 (1.6 mg/ml) mice. In conclusion, our results suggest that transgenic mice can be continuously passed their transgenes to the progeny through the breeding program with the same productivity of human IL-10 protein in their milk.

• Biomolecules & Therapeutics
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• v.30 no.6
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• pp.540-545
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• 2022

Betulin is a triterpenoid natural product contained in several medicinal plants including Betulae Cortex. These medicinal plants have been used for controlling diverse inflammatory diseases in folk medicine and betulin showed anti-inflammatory, antioxidative, and anticancer activities. In this study, we tried to examine whether betulin exerts a regulative effect on the gene expression of MUC5AC mucin under the status simulating a pulmonary inflammation, in human airway epithelial cells. Confluent NCI-H292 cells were pretreated with betulin for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h or the indicated periods. The MUC5AC mucin mRNA expression and mucin glycoprotein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. To elucidate the action mechanism of betulin, effect of betulin on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated by western blot analysis. The results were as follows: 1) Betulin significantly suppressed the production of MUC5AC mucin glycoprotein and down-regulated MUC5AC mRNA expression induced by PMA in NCI-H292 cells. 2) Betulin inhibited NF-κB activation stimulated by PMA. Suppression of inhibitory kappa B kinase (IKK) by betulin led to the inhibition of the phosphorylation and degradation of inhibitory kappa B alpha (IκBα), and the nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of MUC5AC glycoprotein production in NCI-H292 cells. These results suggest betulin inhibits the gene expression of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.

• Journal of Food Hygiene and Safety
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• v.13 no.1
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• pp.6-13
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• 1998

This study was carried out to investigate the antioxidative activity on oxidaton of human low density lipoprotein from marine microorganisms. Bacillus sp. RH-5 producing antioxidative activity have been isolated and identified from coast sea in Pusan. Bacillus sp. RH-5 produced at highest level of antioxidative activity in the medium of 1.0% glucose, 0.25% polypepton, 0.25% yeast extract, 0.01% $FeSO_4{\cdot}7H_2O$ and 50% sea water. The optimal medium pH, cultural temperature and shaking time for the highest production as the antioxidant were 7.0, $30^{\circ}C$ and 48 hr, respectively. The culture broth inhibited the copper catalyzed oxidation of human low density lipoprotein (LDL) at the concentration of 500 and $1,000\;\mu\textrm{g}/ml$ ethylacetate extracts in the presence of $5\;\mu\textrm{M}\;CuSo_4$. The electrophoretic mobility of the LDL oxidized in the presense of $5\;\mu\textrm{M}\;CuSo_4$ was higher than that of native LDL.

• Archives of Pharmacal Research
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• v.29 no.4
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• pp.339-342
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• 2006

A simple high performance liquid chromatographic (HPLC) method was developed for the determination of tolperisone in human plasma. Tolperisone and internal standard (chlorphenesin) were isolated from 1 mL of plasma using 8 mL of dichlormethane. The organic phase was collected and evaporated under nitrogen gas. The residue was then reconstituted with 300 mL aliquot of mobile phase and a 100 mL aliquot was injected onto the $C_{18}$ reverse-phased column. The mobile phase, $45\%$ methanol containing $1\%$ glacial acetic acid and $0.05\%$ 1-hexanesulfonic acid was run at a flow rate of 1 mL/min. The column effluent was monitored using UV detector at 260 nm. The retention times for tolperisone and the internal standard were approximately 7.1 and 8.4 min, respectively. The standard curve was linear with minimal intra-day and inter-day variability. The quantification limit of tolperisone in human plasma was 10 ng/ mL. The proposed method has been applied to the determination of pharmacokinetic profile of tolperisone in Koreans. The T max of tolperisone in Koreans $(0.94{\pm}0.42\;h)$ was not significantly differ from that reported in Europeans (0.5-1 h), but the mean half-life in Koreans $(1.14{\pm}0.27\;h)$ was shorter than that in Europeans $(2.56{\pm}0.2\;h)$. The proposed HPLC method is simple, accurate, reproducible and suitable for pharmacokinetic study of tolperisone.

• BMB Reports
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• v.39 no.4
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• pp.448-451
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• 2006

Chemopreventive and cytotoxic effect of genistein against human breast cancer cell lines was investigated. Genistein inhibited cell proliferation in estrogen receptor-positive (MCF-7) and estrogen receptor-negative (MDA-MB-231) human breast carcinoma cell lines. Cytochrome P450 (CYP) 1A1-mediated ethoxyresorufin O-deethylase (EROD) activity was inhibited by genistein in a concentrationdependent manner. Genistein significantly inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cyclooxy-genase-2 activity and protein expression at the concentrations of 10 (p < 0.05), 25 (p < 0.05) and 50 mM (p < 0.01). In addition, ornithine decarboxylase (ODC) activity was reduced to 53.8 % of the control after 6 h treatment with 50 mM genistein in MCF-7 breast cancer cells. These results suggest that genistein could be of therapeutic value in preventing human breast cancer.

• The Journal of Korean Medicine
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• v.27 no.3 s.67
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• pp.88-106
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• 2006

Objectives: This study was carried out to find the immune responses of the Gami-sopunghwalhyeol-tang $(Ji{\={a}}w{\`{e}}i-sh{\={u}}f{\={e}}nghu{\'{o}}xu{\`{e}}-tang)$ (hereinafter referred to GSHT) to the human fibroblast-like synoviocytes (hFLSs) isolated from patients with rheumatoid arthritis. Methods: Experiments were performed to measure the cytotoxity against hFCs and the production of pro-inflammatory cytokines in hFLSs and the production of NO, ROS. Results: 1. The gene expression of TNF-a, IL-6, IL-8 in hFLSs was effectively reduced at $100{\mu}g/ml$, whereas IL-1 $\beta$ was effectively reduced at 100 and $10{\mu}g/ml$ of GSHT. 2. The gene expression of ICAM-1, MMP-3 in hFLSs was effectively inhibited at 100 and $10{\mu}g/ml$ of GSHT, whereas TIMP-1 was effectively increased at 100 and $10{\mu}g/ml$ of GSHT. 3. The gene expression of NOS-II in hFLSs was effectively inhibited at $100{\mu}g/ml$ of GSHT. 4. The production of NO and ROS in hFLSs was inhibited at 100 and $10{\mu}g/ml$ of GSHT. 5. The proliferation of hFLSs was significantly inhibited at $100{\mu}g/ml$ of GSHT. Conclusions: Comparison of the results for this study showed that Gami-sopunghwalhyeol-tang ($Ji{\={a}}w{\`{e}}i-sh{\={u}}f{\={e}}nghu{\'{o}}xu{\`{e}}-tang$: GSHT) had immunomodulatory effects of suppressing or enhancing.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.263-268
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• 2021

7-Hydroxy-4-methylcoumarin (7H-4MC) inhibits hyaluronan production in multiple cell lines and tissue types both in vitro and in vivo. It is a commercially available drug approved for human use, called hymecromone, in European and Asian countries to prevent biliary spasms. Nevertheless, as the pharmacological efficacy of 7H-4MC has not yet been reported in macrophages, this study investigated its anti-inflammatory effects and mechanism of action using lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. LPS-induced RAW 264.7 cells were treated with various concentrations of 7H-4MC (62.5, 125, 250, and 500 μM). The application of 7H-4MC significantly reduced nitric oxide and prostaglandin E₂ production without cytotoxic effects. Additionally, 7H-4MC strongly decreased the expression of inducible nitric oxide synthase and cyclooxygenase. Furthermore, 7H-4MC reduced the production of proinflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. Finally, 7H-4MC exerted its potent anti-inflammatory actions via the upregulation of IκB-α production, which led to the inhibition of nuclear factor-κB (NF-κB) activity. These results, obtained in macrophage cell lines, suggest that 7H-4MC prevents inflammatory diseases via the NF-κB signaling pathway and that its use could be beneficial for human health. Ultimately, this is the first report describing the anti-inflammatory activity of 7H-4MC in a macrophage cell line.

• The Journal of Korean Medicine
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• v.25 no.3
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• pp.191-202
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• 2004

Objectives : This study was designed to elucidate the synergistic cytotoxic mechanisms of the co-treatment of Gagamjengac-tang (GGJAT) and As₂O₃ in human lung cancer cell line, H-157. Methods : The combination of GGJAT and As₂O₃ synergistically augmented the cytotoxicity of GGJAT and As₂O₃ in H­157 cells. The cytotoxicity by the combination of these two drugs was revealed as apoptosis which was characterized by chromatin condensation and fragmentation in DAPI staining. Results : Antioxidant NAC completely blocked the apoptotic death of H-157 cells by GGJAT and As₂O₃. The apoptotic cytotoxicity of GGJAT and As₂O₃ was accompanied by the induction of DR4 and DR5 in RT-PCR. In addition, antioxidant enzymes such as SOD1, GSH synthetase and GSH reductase were also increased in H-157 cells treated with GGJAT and As₂O₃. However, of note, p53, Fas, FasL and TRAIL were not detected in H-157 cells treated with GGJAT and As₂O₃ by RT-PCR. Conclusions : These results suggest that the synergistic cytotoxicity of the co-treatment of H-457 cells treated with GGJAT and As₂O₃ may cause induction of death receptors DR4 and DR5 as well as reactive oxygen species.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.79-79
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• 2003

Embryonic stem cells are capable of differentiating into a variety of cell lineages. However, the ultimate results of differentiation in vitro greatly depend on the duration of treatment and kinds of differentiating inducers added. In order to investigate the efficiencies of various differentiation inducers and the methods of treatment, we examined differentiation patterns of human embryonic stem cell (hESC, MB03) according to several different protocols. Exp. I) Upon differentiation using retinoic acid and ascorbic acid (RA/AA), embryoid bodies (EB, for 4days) derived from hESC was exposed to Rh (10$^{-6}$ M) and AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. Exp. III) In addition, to examine the effects of neurotrophic factors in the production of mature neurons, groups of cells were exposed to either BDNF (5 ng/ml) or TGF-$\alpha$(10 ng/ml) during the 28 days of final differentiation. Differentiation patterns of RA/AA or bFGF treated groups were very similar; approximately 82% and 83% of the cells, respectively, were positive for anti-NF200 antibody, while it was about 10% and 11%, respectively, for anti-NF160 antibody in 28 days in N2 medium. Alsor, cells expressing TH were as low as 5%, while the cells doubled when matured at the presence of either BDNF or TGF-$\alpha$. Cells immunoreactive to anti-GAD antibody were approximately 20%. These results suggest that a maturation step rather than differentiation induction step, which is formation of EB, effects more decisively to the ultimate differentiation pattern.

• Toxicological Research
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• v.17 no.2
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• pp.131-137
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• 2001

Baicalein, a major flavonoid of extract from Scutellaria baicalensis Georgi, has been shown to exhibit antioxidant and anti proliferative effects. In the present study, we investigate the effects of baicalein on viability and induction of apoptosis in human promyelocytic leukemia HL-60 cells. Baicalein was found to induce apoptosis of HL-60 cells in a concentration-dependent and time-dependent manner. When HL-60 cells were exposed to 100 $\mu\textrm{M}$ baicalein for 6h, the viability was decreased remarkably to 27% of control, whereas DNA fragmentation was significantly increased to 64%. Nucleosomal fragmentation of baicalein treated HL-60 cells, a hallmark of apoptosis, was further identified by agarose gel electrophoresis (DNA ladder). Flow cytometric analysis showed that apoptotic cells were increased to 66.6% after treatment with 100 $\mu\textrm{M}$ baicalein for 6 h. Baicalein-induced apoptosis of HL-60 cells was reduced by 1h pretreatment with inhibitor of caspases, z-Asp-$CH_2$-DCB. At 3 and 10 $\mu\textrm{M}$ of z-Asp-$CH_2$-DCB, DNA fragmentation of HL-60 cells induced by baicalein (50 $\mu\textrm{M}$) was 36.8 and 17.1 %, respectively, whereas, that of HL-60 cells treated by baicalein (50 $\mu\textrm{M}$) without pretreatment with inhibitor of caspases was 62.7%. These data suggest that baicalein induces apoptosis in human leukemia HL-60 cells, and that caspase enzymes might be involved in baicalein-induced apoptosis.