• Journal of Ginseng Research
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• v.39 no.1
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• pp.61-68
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• 2015

Background: Korean Red Ginseng (KRG) is a representative traditional herbal medicine with many different pharmacological properties including anticancer, anti-atherosclerosis, anti-diabetes, and anti-inflammatory activities. Only a few studies have explored the molecular mechanism of KRG-mediated anti-inflammatory activity. Methods: We investigated the anti-inflammatory mechanisms of the protopanaxadiol saponin fraction (PPD-SF) of KRG using in vitro and in vivo inflammatory models. Results: PPD-SF dose-dependently diminished the release of inflammatory mediators [nitric oxide (NO), tumor necrosis factor-${\alpha}$, and prostaglandin $E_2$], and downregulated the mRNA expression of their corresponding genes (inducible NO synthase, tumor necrosis factor-${\alpha}$, and cyclooxygenase-2), without altering cell viability. The PPD-SF-mediated suppression of these events appeared to be regulated by a blockade of p38, c-Jun N-terminal kinase (JNK), and TANK (TRAF family member-associated NF-kappa-B activator)-binding kinase 1 (TBK1), which are linked to the activation of activating transcription factor 2 (ATF2) and interferon regulatory transcription factor 3 (IRF3). Moreover, this fraction also ameliorated HCl/ethanol/-induced gastritis via suppression of phospho-JNK2 levels. Conclusion: These results strongly suggest that the anti-inflammatory action of PPD-SF could be mediated by a reduction in the activation of p38-, JNK2-, and TANK-binding-kinase-1-linked pathways and their corresponding transcription factors (ATF2 and IRF3).

• Neonatal Medicine
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• v.17 no.2
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• pp.181-192
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• 2010

Purpose: Current studies have demonstrated the neuroprotective effects of dizocilpine (MK-801) in many animal models of brain injury, including hypoxic-ischemic (HI) encephlopathy, trauma and excitotoxicity, but limited data are available for those during the neonatal periods. Here we investigated whether dizocilpine can protect the developing rat brain from HI injury via anti-apoptosis. Methods: In an in vitro model, embryonic cortical neuronal cell culture of Sprague-Dawley (SD) rats at 18-day gestation was done. The cultured cells were divided into three groups: normoxia (N), hypoxia (H), and hypoxia treated with dizocilpine (HD). The N group was prepared in 5% $CO_2$ incubators and the other groups were placed in 1% $O_2$ incubators (94% N2, 5% $CO_2$) for 16 hours. In an in vivo model, left carotid artery ligation was done in 7-day-old SD rat pups. The animals were divided into six groups; hypoxia (N), hypoxia (H), hypoxia with sham-operation (HS), hypoxia with operation (HO), HO treated with vehicle (HV), and HO treated with dizocilpine (HD). Hypoxia was made by exposure to a 2 hour period of hypoxic incubator (92% N2, 8% $O_2$). Results: In the in vitvo and in vivo models, the expressions of Bcl-2 in the hypoxia groups were reduced compared to the normoxia group. whereas those in the dizocilpine-treated group were increased compared to the hypoxia group. However. the expressions of Bax and caspase-3 and the ratio of Bax/Bcl-2 were revealed reversely. Conclusion: Dizocilpine has neuroprotective property over perinatal HI brain injury via anti-apoptosis.

• Journal of Preventive Medicine and Public Health
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• v.27 no.1 s.45
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• pp.11-24
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• 1994

The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

• The Journal of Korean Obstetrics and Gynecology
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• v.30 no.2
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• pp.49-70
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• 2017

Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.

• Korean Journal of Food Science and Technology
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• v.38 no.2
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• pp.290-294
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• 2006

Inactivation rates of Salmonella enteritidis in vitro and in vivo were assessed using confocal microscopy and flow cytometry. S. enteritidis was inactivated with 1% (w/v) trisodium phosphate (TSP) and live cells, and inactive cells were distinguished by staining with fluorescent probe, LIVE/DEAD BacLight Bacteria Viability stain. After TSP treatment for 1 min, most of Salmonella cells changed from green (live cells) fluorescence to red (inactive cells) fluorescence, indication of effective sanitizing. Inactivation efficiency and contamination sites of S. enteritidis on chicken skin by TSP treatment were assessed using confocal laser microscopy. Precise flow cytometry histograms for viability changes of S. enteritidis. after TSP treatments were obtained. Efficiency of various sanitizer treatments on foodborne pathogens could be assessed using this method.

• Journal of Life Science
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• v.23 no.9
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• pp.1147-1154
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• 2013

This study was conducted to evaluate the antioxidation activity of Jeju crossbred horse (Jeju native horse ${\times}$ thoroughbred) leg bone extracts (HLBE) and its enzyme hydrolysates by determination of 1,1-diphenyl-2picryl-hydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzo thiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity, ferric reducing/antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC). HLBE was extracted with hot water for 24 hr and lyophilized. The lyophilized HLBE was hydrolyzed using multifect PR6L (MP), pepsin (PS), and a pepsin and pancreatin mixture (PSPC) for 4 hr at 60, 50, and $50^{\circ}C$, respectively. The hydrolysates were separated by a molecular weight of 3 kDa more or less. When the yield of HLBE was 100%, the yield of hydrolysates less than 3 kDa of MP, PS, and PSPC was 10.86, 3.26, and 8.00%, respectively. Enzyme hydrolysates with low molecular weight less than 3 kDa in MP and PSPC showed significantly higher DPPH, ABTS radical scavenging activity, and ORAC compared to HLBE and its hydrolysates with more than 3 kDa. However, the FRAP of the hydrolysates less than 3 kDa in PS was significantly higher than in MP. These results suggest that low molecular weight enzyme hydrolysates less than 3 kDa have more powerful antioxidation activity, especially when they are hydrolyzed by MP and PSPC rather than PS. Therefore, low molecular weight enzyme hydrolysates of HLBE hydrolyzed with MP and PSPC have significant potential as antioxidants in the food industry. Further in vivo studies are required to support the antioxidation activities of the hydrolysates in vitro.

• Journal of Oral Medicine and Pain
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• v.34 no.3
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• pp.261-276
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• 2009

Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

• Journal of The Korean Society of Grassland and Forage Science
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• v.5 no.2
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• pp.127-135
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• 1985

The effects of morphological development and environmental temperature on synthesis and accumulation behavior of cell-wall constituents were studied in maize cv. Blizzard and sorghum cv. Sioux and Pioneer 931 at Muenchen Technical University from 1979 to 1981. Various growth stages of maize and sorghum plants were grown on field and phytotron at 4 temperature regimes of 30/25, 25/20, 28/18 and 18/8 degree C and mid-summer sunlight over 13-hour days. The results are summarized as follow: 1. Cell-wall constituents in sorghum and maize plants were shown to have a great synthesis rates at early growth stage from growing point differentiation to final leaf visible. The highest concentration of cell wall contents were found at heading stage with 52-54% and 64-68% of neutral detergence fiber, and 30% and 45% of acid detergence fiber foe maize and sorghum, respectively. 2. The structural carbohydrates, cellulose and hemicellulose, were found as a main components of cell-wall constituents. Cellulose were mainly accumulated in stalks, while hemicellulose were an important cell wall components in leaves and panicle. 3. Synthesis rates of cell-wall constituents and non-strnctural carbohydrates were associated with increasing of temperature. Reserved carbohydrates such as fructosan, mono - and dissaccharose in plant were, however, declined when the temperature exceeded 30 deg C, during the accumulation of cellulose, hemicellulose and lignin were increased continuously. 4. Cell-wall constituents lowered digestibility and net energy accumulation in sorghum and maize plants. In a in vitro and in vivo trial, it was found a negative correlation between digestion dry matter and cell wall constituents, especially cellulose and lignin.

• Journal of Life Science
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• v.19 no.6
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• pp.770-780
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• 2009

A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a), was derived from Spirulina platensis. We conducted a series of experiments, in vitro and in vivo, to evaluate the anticancer effect and mechanism of photodynamic therapy using 9-HpbD-a and 660 nm diode lasers on a squamous carcinoma cell line. We studied the cytotoxic effects of pheophytin-a, 9-HpbD-a, 9-HpbD-a red and 660 nm diode lasers in a human head and neck cancer cell line (SNU-1041). Cell growth inhibition was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of 9-HpbD was higher than those of 9-HpbD-a red or pheophytin-a in PDT. We then tested the cytotoxic effects of 9-hydroxypheophorbide-a (9-HpbD-a) in vitro. The cultured SNU-I041 cells were treated with serial concentrations of 9-HpbD-a followed by various energy doses (0, 0.1, 0.5, 3.2 J/$cm^{2}$) and by various interval times (0, 3, 6, 9, 12 hr) until laser irradiation, then MTT assay was applied to measure the relative inhibitory effects of photodynamic therapy (PDT). Optimal laser irradiation time was 30 minutes and the cytotoxic effects according to incubation time after 9-HpbD-a treatment increased until 6 hours, after which it then showed no increase. To observe the cell death mechanism after PDT, SUN-I041 cells were stained by Hoechst 33342 and propidium iodide after PDT, and observed under transmission electron microscopy (TEM). The principal mechanism of PDT at a low dose of 9-HpbD-a was apoptosis, and at a high dose of 9-HpbD-a it was necrosis. PDT effects were also observed in a xenografted nude mouse model. Group I (no 9-HpbD-a, no laser irradiation) and Group II (9-HpbD-a injection only) showed no response (4/4, 100%), and Group III (laser irradiation only) showed recurrence (1/4,25%) or no response (3/4, 75 %). Group IV (9-HpbD-a + laser irradiation) showed complete response (10/16, 62.5%), recurrence (4/16, 25%) or no response (2/16, 12.5%). Group IV showed a significant remission rate compared to other groups (p<0.05). These results suggest that 9-HpbD-a is a promising photosensitizer for the future and that further studies on biodistribution, toxicity and mechanism of action would be needed to use 9-HpbD-a as a photosensitizer in the clinical setting.

• Korean Journal of Food Science and Technology
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• v.38 no.1
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• pp.104-108
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• 2006

Water extracts of Jehotang were evaluated for their growth-promoting effects on Bifidobacterium longum, Lactobacillus sp., L. acidophilus, and Clostridium perfringens. Addition of Jehotang water extract to modified EG media at 0.1 mg/mL increased growths of B. longum, Lactobacillus sp., and L. acidophilus, with 1.8-fold increase in growth of L. acidophilus compared to that of control. Studies on these strains by agar diffusion method showed Lactobacillus sp. and L. acidophilus were activated by addition of Jehotang extract at 10 mg/disc. Proliferation responses of mice splenocytes and Peyer's patch cells to ConA by LPS-stimulation at 500 mg/kg B.W./day Jehotang extract were investigated in vitro. Upon treatment of 1 mg/mL Jehotang water extract to mice, proliferations of splenocytes and Peyer's patch cells increased 1.4- and 1.6-fold compared to control, respectively. In mice administered Jehotang extract, production of intestinal secretory IgA (sIgA) increased 2.4-fold compared to control. These results indicate water extract of Jehotang stimulated intestinal immune system of mice. In mice treated with Jehotang extract, production of lymphocytes was 4% lower, whereas those of granulocytes and platelets were 4% and slightly higher than control, respectively.