• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6261-6265
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• 2013

Background: Recent studies have suggested that expression of the RAS protein activator like-1 gene (RASAL1) is decreased in gastric carcinoma tissues and cell lines, indicated a role in tumorigenesis and development of gastric cancer. Reduced expression of RASAL1 could result in aberrant increase of activity of RAS signaling pathways in cancer cells. However, the exact mechanism which induces down-regulation of the RASAL1 gene remains unclear. This study aimed to determine the methylation status and regulation of RASAL1 in gastric cancer. Materials and Methods: Using the methylation-specific polymerase chain reaction (MSP), the methylation status of CpG islands in the RASAL1 promoter in gastric cancers and paired adjacent non-cancerous tissues from 40 patients was assessed and its clinicopathological significance was analyzed. The methylation status of RASAL1 in gastric cancer lines MKN-28, SGC-790l, BGC-823, as well as in normal gastric epithelial cell line GES-l was also determined after treatment with a DNA methyltransferase inhibitor, 5-aza-2'-doexycytidine (5-Aza-CdR). RAS activity (GAS-GTP) was assessed through a pull-down method, while protein levels of ERK1/2, a downstream molecule of RAS signaling pathways, were determined by Western blotting. Results: The frequencies of RASAL1 promoter methylation in gastric cancer and paired adjacent non-cancerous tissues were 70% (28/40) and 30% (12/40) respectively (P<0.05). There were significantly correlations between RASAL1 promoter methylation with tumor differentiation, tumor size, invasive depth and lymph node metastasis in patients with gastric cancer (all P<0.05), but no correlation was found for age or gender. Promoter hypermethylation of the RASAL1 gene was detected in MKN-28, SGC-790l and BGC-823 cancer cells, but not in the normal gastric epithelial cell line GES-1. Elevated expression of the RASAL1 protein, a decreased RAS-GTP and p-ERK1/2 protein were detected in three gastric cancer cell lines after treatment with 5-Aza-CdR. Conclusions: Aberrant hypermethylation of the RASAL1 gene promoter frequently occurs in gastric cancer tissues and cells. In addition, the demethylating agent 5-Aza-CdR can reverse the hypermethylation of RASAL1 gene and up-regulate the expression of RASAL1 significantly in gastric cancer cells in vivo. Our study suggests that RASAL1 promoter methylation may have a certain relationship with the reduced RASAL1 expression in gastric cancer.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.7
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• pp.895-900
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• 2012

The effects of three fractions, hexane (BHHH), chloroform (BHHC), and ethyl acetate (BHHE), from water extract of Benincasa hispida on the underlying mechanisms of adipogenesis were investigated in 3T3-L1 cells. Intracellular lipid droplets were stained with Oil Red O dye and quantified. Compared to control, lipid accumulation significantly decreased by 11% and 13% upon treatment with BHHC and BHHE, respectively at a concentration of 50 ${\mu}g/mL$. Intracellular triglyceride (TG) levels were also reduced by 21% and 16%, respectively, at the same concentration. To determine the mechanism behind the reductions in TG content and lipid accumulation, glycerol release and expression levels of adipogenic marker genes were measured. The levels of free glycerol released into culture medium increased by 13% and 17% upon treatment with BHHC and BHHE, respectively. In subsequent measurements using real-time polymerization chain reaction, the mRNA levels of $PPAR{\gamma}$, C/$EBP{\alpha}$, and leptin significantly decreased upon treatment with BHHE (45%, 67%, and 35%) in comparison with non-treated control. These results suggest that BHHE inhibits adipocyte differentiation by blocking $PPAR{\gamma}$, C/$EBP{\alpha}$, and leptin gene expression in 3T3-L1 cells, resulting in reduced lipid accumulation, increased glycerol release, and intracellular triglycerides.

• Animal Bioscience
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• v.37 no.5
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• pp.929-943
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• 2024

Objective: The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis. Methods: The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their negative control (NC) in bovine adipocytes, and the multiplicity of infection, transfection efficiency, interference efficiency were evaluated through quantitative real-time polymerase chain reaction, western blotting and fluorescence microscopy. The lipid droplets, triglycerides (TG) content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing. Results: The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their down-regulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as CCAAT enhancer binding protein alpha (C/EBPα), fatty acid binding protein (FABP4), peroxisome proliferator-activated receptor-γ (PPARγ), lipoprotein lipase (LPL), and Fas cell surface death receptor (FAS), similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPα, PPARγ, FABP4, and LPL genes (p<0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1,774 differentially expressed genes (DEGs) including 157 up regulated and 1,617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top Kyoto encyclopedia of genes and genomes pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes. Conclusion: Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection.

• Journal of Animal Environmental Science
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• v.11 no.3
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• pp.207-214
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• 2005

To reduce phosphorus and nitrogen from the swine wastewater, magnesium chloride $(MgCl_2)$ was used as a reaction material for both soluble phosphorus (SP) and ammonia-nitrogen (AN). The initial value of SP content were $471mg/\ell$ far aeration test and $515 mg/\ell$ for NaOH addition test, but treatment of $MgCl_2$ reduced SP value to $5mg/\ell$ and $4mg/\ell$. The removal efficiency of $MgCl_2$ for SP showed $99\%$ in both treatment, and the removal efficiency of $MgCl_2$ for AN showed $15\%$ with treatment of aeration and $18\%$ with NaOH. All the experiments were done in a low temperature of 6 to $8^{\circ}C$, suggesting that this methods are possibly able to apply to a cold weather conditions. Moreover, the struvite crystal structure was identified by electronic microscope, implying that $MgCl_2$ is an effective material for removal of SP from swine wastewater In addition to the increased removal rate of the AN in wastewater, both $MgCl_2$ and $KH_2PO_4$ were added. The SP value was reduced by $99\%$ with 2g addition of the phosphate. The SP removal rate by 4g addition of the phosphate was increased only as $15-19\%$, but the quantity of removed SP was higher than that of 2g addition test. The value of AN was not reduced as expected by adding $KH_2PO_4$. The AN removal rate were low as $18\%$ and $15\%$ like as the level of the former test with $MgCl_2$ alone. Therefore, it is needed to examine closely the reaction mechanism f3r reducing both SP and AN simultaneously.

• Economic and Environmental Geology
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• v.36 no.1
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• pp.39-48
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• 2003

The removal of chromate from aqueous solution using finely ground pyrite and biotite was investigated by batch experiments and the kinetics and the mechanism of chromate reduction were discussed. The chromate reduction by pyrite was about hundred times faster than that by biotite and was also faster at pH 3 than at pH 4. When pyrite was used, more than 90% of initial chromate was reduced within four hours at pH 4 and within 40 min. at pH 3. However, more than 400 hours was taken for the reduction of 90% of initial chromate by biotite. The results indicate that the rate of chromate reduction was strongly depending on the amount of Fe(II) in the minerals and on the dissolution rate of Fe(II) from the minerals. The reduction of chromate at pH 4 resulted in the precipitation of (Cr, Fe)(OH))$_3$$_{ (s)}$, which is believed to have limited the concentrations of dissolved Cr(III) and Fe(III) to less than expected values. When biotite was used, amounts of decreased Fe(II) and reduced Cr(Ⅵ) did not show stoichiometric relationship, which implying there was not only chromate reduction by ferrous ions in the acidic solution but also heterogeneous reduction of ferric ions by the structural ferrous iron in biotite. However, the results from a series of the experiments using Pyrite showed that concentrations of the decreased Fe(II) and the reduced Cr(Ⅵ) were close to the stoichiometric ratio of 3:1. It was because the oxidation of pyrite rapidly created ferrous ions even in oxygenated solutions and the chromate reduction by the ferrous ions was significantly faster than ferrous ion oxygenation.

• Journal of the korean academy of Pediatric Dentistry
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• v.23 no.3
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• pp.640-658
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• 1996

The purpose of this study was to examine the mechanism of improving acid resistance of Nd-YAG laser irradiated tooth enamel and determine the most effective energy density for improving acid resistance. The bovine tooth enamel were lased with a pulsed Nd-YAG laser. The energy densities of exposed laser beam were varied from 10 to $70\;J/cm^2$. To investigate the degree of improving acid resistance by irradiation, all the samples were submerged to demineralize in 0.5 N $HClO_4$ solution for 1 minute. After 1 minute, 0.05 % $LaCl_3$ was added to the solution for interrupting the demineralization reaction. The amounts of dissolved calcium and phosphate in the solution were measured by using an atomic absorption spectrophotometer and the UV/VIS spectrophotometer, respectively. To examine the mechanism of improving acid resistance, X-ray diffraction analysis, infrared spectroscopy, and scanning electron microscopy were taken. The X-ray diffraction pattern of the samples were obtained in the $10^{\circ}{\sim}80^{\circ}2{\theta}$ range with $Cu-K{\alpha}$ radiation using M18HF(Mac Science Co.) with X-ray diffractometer operating at 40 KV and 300 mA. The infra-red spectra of the ground samples in 300 mg KBr pellets 10 mm diameter were obtained in the $4000cm^{-1}\;to\;400cm^{-1}$ range using JASCO 300E spectrophotometer. The scanning electron microscopy was carried out using JSM6400(JEOL Co.) with $500{\sim}2000$ times magnification. The results were as follow 1. The concentration of calcium dissolved from laser irradiated enamel with $50J/cm^2$ was significantly lesser than that of unlased control group (p<0.05) 2. From the result of the X-ray diffraction analysis, $\beta$-TCP, which increases acid solubility, was identified in lased enamel but the diffraction peaks of (002) and (004) became sharp with increasing energy density of laser irradiation. This means that the crystals in lased samples were grown through the c-axis and subsequently, the acid solubility of enamel decreased. 3. The a-axis parameter was slightly increased by laser irradiation, whereas the c-axis parameter was almost constant except for a little decrease at $50J/cm^2$. 4. In the infra-red spectra of lased enamels, phosphate bands ($600{\sim}500cm^{-1}$), B-carbonate bands (870, $1415{\sim}1455cm^{-1}$), and A-carbonate band ($1545cm^{-1}$) were observed. The amounts of phosphate bands and the B-carbonate bands were reduced, on the other hand, the amount of the A-carbonate band was increased by increase the energy density. 5. The SEM experiments reveal that the surface melting and recrystallization were appeared at $30J/cm^2$ and the cracks were observed at $70J/cm^2$. From above results, It may be suggested that the most effective energy density for improving acid resistance of tooth enamel with the irradiation of Nd-YAG laser was $50J/cm^2$. The mechanism of improving acid resistance were reduction of permeability due to surface melting and recrystallization of lased enamel and reduction of acid solubility of enamel due to decrease of carbonate content and growth of crystal.

• Journal of the Korean Chemical Society
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• v.33 no.3
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• pp.273-280
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• 1989

The spectra of the $Co^{II}CyDTA$(CyDTA: cyclohexyldiaminetetraacetic acid) complex have been measured in aqueous solution of pH = 6-13.2. The red shift of the spectrum in the more basic solution was ascribed to the transformation of $CoCyDTA^{2-}$ into $CoCyDTA(OH)^{3-}$. The equilibrium constant, $K_{OH} = [CoCyDTA(OH)^{3-}]/[CoCyDTA^{2-}][OH^-]$ was $75M^{-1}$ at $40^{\circ}C$. The electron transfer reactions of $CoCyDTA^{2-}$ and $CoCyDTA(OH)^{3-}$ with $Fe(CN)_6^{3-}$ have been studied using spectrophotometric technique in the range of pH applied to the determination of equilibrium constant. The pseudo first-order rate constants observed ($k_{obs}$) were not changed upto pH = 10.8, but increased with increasing pH in the range of pH = $10.8{\sim}13.0$. The rate law reduced in the range of pH = 6-13 was $k_{obs} = (k_3[CoCyDTA^{2-}] + k_4[CoCyDTA(OH)^{3-}])/(1+K_1[CoCyDTA^{2-}])$. The rate constants of the reactions (3a) and (3b), $k_3$ and $k_4$ respectively have been determined to be 0.529 and $4.500M^{-1}sec^{-1}$ at $40^{\circ}C$. The activation entropies (147{\pm}1.1JK^{-1} mol^{-1}$ at pH = 10.8) and activation volumes $(6.25cm^3mol^{-1}, pH = 10.8)$ increased with increasing pH, while the activation enthalpy (12.44 ${\pm}$ 0.20 kcal/mole) was independent of pH. Using the pH effect on the rate constants, the activation entropies and the activation volumes, the mechanism of the electron transfer reaction for $Co^{II}-Fe^{III}$ system was discussed.

• The Journal of Pediatrics of Korean Medicine
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• v.32 no.3
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• pp.1-15
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• 2018

Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.3
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• pp.419-427
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• 2016

There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at $42^{\circ}C$ for one hour and then allowed to recover at normal incubation temperature of $37^{\circ}C$ for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to $400{\mu}g/mL$) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.

• Journal of Korean Neurosurgical Society
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• v.47 no.5
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• pp.345-351
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• 2010

Objective : In this study, the Mindfulness Based Stress Reduction (MBSR) program was applied to patients presenting with depression and anxiety after surgery from spontaneous subarachnoid hemorrhage (SAH) and the effects were assessed. Methods : The subjects were patients admitted for cerebral aneurysm rupture and treated by means of surgery from March to December, 2007. More than 6 months had passed after surgery, without any special lesions showing up on computed tomography (CT), and the Glasgow outcome scale (GOS) was 5 points. Among patients with anxiety and depression symptoms, 11 patients completed the program. The MBSR program was conducted once a week, 2.5 hours each, for 8 weeks. The evaluation criteria were : 1) the Beck Depression Inventory (BDI): it measures the type and level of depression, 2) the State-Trait Anxiety Inventory : the anxiety state of normal adults without mental disorder, and 3) Heart Rate Variability (HRV) : the influence of the autonomous nervous system on the sinoarterial node varies continuously in response to the change of the internal/external environment. Results : The BDI value was decreased from 18.5 ${\pm}$ 10.9 to 9.5 ${\pm}$ 7.1 (p = 0.013) : it was statistically significant, and the depression level of patients was lowered. The state anxiety was decreased from 51.3 ${\pm}$ 13.9 to 42.3 ${\pm}$ 15.2; the trait anxiety was reduced from 50.9 ${\pm}$ 12.3 to 41.3 ${\pm}$ 12.8, and a borderline significant difference was shown (p = 0.091, p = 0.056). In other words, after the treatment, although it was not statistically significant, a decreased tendency in anxiety was shown. In the HRV measurement, standard deviation normal to normal (SDNN), square root of the square root of the mean sum of squared differences between adjacent normal to normal intervals (RMSSD), and total power (TP) showed significant increase, Physical Stress Index (PSI) showed a significant reduction, and thus an improvement in the homeostatic control mechanism of the autonomic nervous system was ween. Conclusion : The MBSR program was applied to the patients showing anxiety and depression reaction after SAH treatment, and a reduction in depression symptoms and physiological reactions were observed. The application of the MBSR program may be considered as a new tool in improving the quality of life for patients after surgery.