• 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 Applied Biological Chemistry
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• v.66
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• pp.369-378
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• 2023

Smilax sieboldii is one of the Smilax species. A number of Smilax plants have multiple physiologically-active components and anti-inflammatory/anti-oxidant effects. Antiobesity effects induced by Smilax sieboldii have not been reported. In this study, we investigated the effects and molecular mechanisms of anti-obesity activity of 70% ethanol Smilax sieboldii extract (SSE). The anti-obesity effect of SSE was determined using 3T3-L1 adipocytes. We confirmed that SSE was not cytotoxic to murine 3T3-L1 preadipocytes, we evaluated SSE dose-dependently decreased the accumulation of lipids via an Oil Red O assay and triglyceride assay. These anti-obesity activities of SSE were mediated by the inhibition of adipogenesis-related marker genes (peroxisome proliferator activated receptor-γ, CCAAT-enhancer-binding protein α, and SREBP1c) and lipogenesis-related marker genes (fatty acid synthase and aP2). These results suggest that SSE has the potential to exert anti-obesity and anti-hyperlipidemia effects by regulating adipogenic transcription factors and inhibiting the expression of adipogenic markers.

• Journal of Korean Medicine for Obesity Research
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• v.24 no.1
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• pp.1-12
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• 2024

Objectives: Ephedrae herba (EH) and Coicis semen (CS) has been frequently prescribed for the treatment of obesity. However, effects of combinational extracts of these two herbs on non-alcoholic fatty liver disease are unknown. The aim of the present study was to investigate the effects of EH and CS on lipid accumulation and glucose absorption in free fatty acids (FFAs) or palmitic acid (PA)-treated HepG2 cells. Methods: Five samples of EH and CS were extracted by combination ratios (S1=0:100, S2=25:75, S3=50:50, S4=75:25, S5=100:0). Oil Red O staining was used to measure lipid accumulation in FFAs-induced steatosis cells. Intracellular triglycerides and total cholesterol levels were measured in FFAs-induced steatotic HepG2 cells. In PA-treated cells, intracellular 2-NBDG was detected using a fluorescence microplate reader and flow cytometry. Phosphorylation of key metabolism-related factors of AMP-activated protein kinase and acetyl-CoA carboxylase, expression of key lipid synthesis-related factors carnitine palmitoyltransferase 1 alpha (CPT1α), sterol regulatory element-binding protein 1 (SREBP1), peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT enhancer binding protein alpha (C/EBPα) were confirmed by western blot. Results: Treatment of EH-CS combination in the FFAs-induced steatotic HepG2 cells significantly reduced lipid accumulation. As the relative ratio of Ephedrae herba increased, the lipid-lowering effects of the combination were increased. However, S1 and S5 of Ephedrae herba and Coicis semen did not significantly reduce triglycerides and total cholesterol induced by FFAs. However, the combination of Ephedrae herba and Coicis semen restored glucose absorption in PA-induced HepG2 cells. Major makers of SREBP1, PPARγ, C/EBPα, and CPT1α expression tended to decrease with EH ratio. Conclusions: The EH-CS combination has advantages over sole EH and CS extracts in improving lipid and glucose metabolism in liver steatosis models.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7179-7188
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• 2015

Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.11
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• pp.1627-1634
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• 2014

The recipe for sujeonggwa, a Korean traditional sweet drink containing cinnamon, ginger, sugar, or honey, was modified by replacing sugar with alternative sweeteners [stevia or short-chain frutooligosaccharide (scFOS)] in order to improve the health functionality of sujeonggwa. The aim of this study was to evaluate the effects of modified sujeonggwa on lipid peroxidation and oxidized DNA damage in diet-induced hypercholesterolemic ApoE knockout mice. Hypercholesterolemia was induced in 6-week-old male mice by administration of a high cholesterol diet (1.25% cholesterol, 0.5% cholic acid, and 10% coconut oil) for 4 weeks, after which mice were divided into five groups: sucrose solution-fed control group, sujeonggwa containing sucrose group, sucrose+stevia group, sucrose+stevia+scFOS group, and commercially available sujeonggwa group as a positive control. After 6 weeks, sujeonggwa supplementation resulted in reduced hepatic thiobarbituric acid reactive substances (TBARS), regardless of sweetener type. However, reduction of hepatic TBARS by commercially available sujeonggwa was insignificant. Both endogenous and $H_2O_2$-induced DNA damage in hepatocytes and splenocytes were significantly reduced only in the sujeonggwa containing stevia group compared to the sucrose-fed control group. There were no significant effects of sujeonggwa supplementation on total radical trapping potential, lipid peroxidation, or DNA damage in blood. These results suggest that sujeonggwa has protective effects against hepatic lipid peroxidation and DNA damage in hepatocytes or splenocytes from diet-induced hypercholesterolemic ApoE knockout mice, and the type of sweetener should be modified to improve the health benefits of sujeonggwa.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.5
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• pp.659-666
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• 2010

Lysobacter antibioticus HS124 was isolated from rhizosphere soil in previous experiments, which produced lytic enzymes such as chitinase, gelatinase, lipase and protease. In addition, HS124 released an antibiotic compound, 4-hydroxyphenylacetic acid (4-HPAA). When larvae of P. xylostella was treated with HS124 culture broth, its body was destroyed, and degraded with the increase of incubation time, yielding glycine which was detected from HS124 culture broth. When 4-HPAA produced from HS124 was sprayed, larvae mortality increased with increasing concentration of 4-HPAA. When HS124 culture supplemented with Tween 80 was sprayed, its insecticidal activity against larvae was approximately 1.4 times higher compared to the culture without Tween 80. Insecticide (IS), HS124 culture broth (HS124), Magic-pi (MP) and HS124 culture broth+Magic-pi (HS124+MP) were each treated against larvae of P. xylostella to investigate their insecticidal effect where sterile diluted water (SDW) was used as a control. The highest mortality of larvae was found in HS124+MP, followed by IS, MP, HS124 and SDW respectively. Mortality of larvae in HS124 was 31% higher than that in SDW, but 41% lower than that in HS124+MP, meaning that both enzymes and antibiotics produced from HS124 may synergistically act as active agents with plant extract containing neem oil and turmeric in HS124+MP treatment. These results suggested that L. antibioticus HS124 together with plant extract can be one of candidates for biocontrol agents against Plutella xylostella.

• Journal of Food Hygiene and Safety
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• v.22 no.4
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• pp.382-387
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• 2007

The approval of use of certain food-grade phosphates as food additives in a wide variety of meat products greatly stimulated research on the applications of phosphates in foods. Phytic acid is a natural plant inositol hexaphosphate constituting 1-5% of most cereals, nuts, legumes, oil seeds, pollen, and spores. In this study, we investigated antibacterial activity of sodium phytate (SPT) against Salmonella typhimurium in tryptic soy broth with different pHs and in chicken, pork and beef. In tryptic soy broth, SPT at the concentrations of 0.1, 0.5, and 1.0% effectively inhibited the growth of Salmonella typhymurium in a concentration-dependent manner. At pH 5.5-7.0 similar to meat pHs, 1% SPT almost completely inhibited the bacterial growth. The inhibitory effect of SPT was stronger at pH 7.0 than pH 5.5. In chicken, pork, and beef, SPT at the concentrations of 0.1, 0.5, and 1% significantly inhibited the growth of Salmonella typhimurium in a dose-dependant manner (p<0.01). The addition of 1% SPT in the meats significantly increased the meat pHs. These results indicate that SPT is very effective for inhibition of bacterial growth as a muscle food additive for increasing food safety and functions.

• Journal of the East Asian Society of Dietary Life
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• v.20 no.1
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• pp.20-29
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• 2010

Naringin has antioxidant and antihyperlipidemic properties, however, phenolic compounds including naringin are unstable in the presence of light, heat and oxygen. Beta-cyclodextrin ($\beta$-CD) is a cyclic heptamer composed of seven glucose units that enhances the stability and solubility of molecules through the formation of inclusion complexes. This study was conducted out to compare the effects of CD-naringin (CD-N) inclusion complexes with naringin on lipid metabolism in high fat-fed animals. Male C57BL/6 mice were fed either CD-N (0.048%, w/w) or naringin (N, 0.02%, w/w) in a 20% high-fat (HFC, 15% lard, 5% corn oil, w/w) diet for 10 weeks. Orlistat (Xenical, 0.01%, w/w) was used as a positive control (PC). There were no differences in body weight, food intake, liver and heart weights, plasma triglyceride(TG), leptin, adiponectin, resistin, IL-$1{\beta}$ and IL-6 concentrations, and hepatic $\beta$-oxidation, carnitine palmitoyl transferase(CPT), glucose-6-phosphate dehydrogenase (G6PD) and malic enzyme activities between the HFC and CD-N groups or between the HFC and N groups. However, both CD-naringin and naringin supplementation les to a significant reduction in the epididymal and perirenal white adipose tissue weights, plasma free fatty acid, insulin and blood glucose concentrations, hepatic cholesterol and TG contents and hepatic fatty acid synthase (FAS), phosphatidate phosphohydrolase (PAP) and HMG-CoA reductase activities compared to the HFC group. The plasma HDL-cholesterol concentration was significantly higher in CD-N and N groups than in HF and PC groups. These results indicate that both CD-naringin and naringin supplementation effectively improved plasma and hepatic lipid metabolism without differences between CD-N and naringin groups.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1015-1021
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• 2013

Glycyrrhiza inflata Batal, an important species of licorice, is one of the most widely used medicinal plants for over 4000 years. Glycyrrhiza plant species has been well known for its various therapeutic activities such as anti-inflammatory, anti-allergic, and anti-ulcer. The purpose of this study was to determine the effects of Glycyrrhiza inflata Batal ethanol extracts (GBE) on adipocyte and osteoblast differentiation. Mesenchymal C3H10T1/2 cells were treated with sub-cytotoxic doses of GBE, and its effects on adipocyte differentiation were assessed. We found that GBE dose-dependently increased lipid accumulation and also induced the expression of adipocyte markers, such as $PPAR{\gamma}$ and its target genes, aP2, and adiponectin, in C3H10T1/2 cells. Consistently, similar effects of GBE on lipid accumulation were also observed in preadipocyte 3T3-L1 cells that further supports the pro-adipogenic activities of GBE. We also investigated the effects of GBE on osteoblast differentiation of mesenchymal C3H10T1/2 cells. As a results, we found that GBE increased the activity of alkaline phosphatase in a dose-dependent manner and also promoted the expression of osteoblast markers, such as ALP and RUNX2, during osteoblast differentiation of C3H10T1/2 cells. Similar pro-osteogenic effects of GBE were also observed in preosteoblast MC3T3-E1 cells. Finally, our data show that a major bioactive compound found in Glycyrrhiza inflata Batal, licochalcone A (LA) but not glycyrrhizic acid (GA), can mediate the pro-adipogenic and pro-osteogenic effects of GBE. Taken together, this study provides data to show the possibility of GBE and its bioactive component LA as putative strategies for type 2 diabetes and bone diseases.

• Journal of Environmental Impact Assessment
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• v.29 no.3
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• pp.157-181
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• 2020

Sediments from rivers, lakes and marine ports serve as end points for pollutants discharged into the water, and at the same time serve as sources of pollutants that are continuously released into the water. Until now, the contaminated sediments have been landfilled or dumped at sea. Landfilling, however, was expensive and dumping at sea was completely banned due to the London Convention. Therefore, this study applied contaminated sedimentation soil of 'Royal Palace Livestock Complex' as soil purification method. Soil remediation methods were applied to pretreatment, composting, soil washing, electrokinetics, and thermal desorption by selecting overseas application cases and domestically applicable application technologies. As a result of surveying the site for pollutant characteristics, Disolved Oxigen (DO), Suspended Solid (SS), Chemical Oxygen Demand (COD), Total Nitrogen (TN), and Total Phosphorus (TP) exceeded the discharged water quality standard, and especially SS, COD, TN, and TP exceeded the standard several tens to several hundred times. Soil showed high concentrations of copper and zinc, which promote the growth of pig feed, and cadmium exceeded 1 standard of Soil Environment Conservation Act. In the pretreatment technology, hydrocyclone was used for particle size separation, and the fine soil was separated by more than 80%. Composting was performed on organic and Total Petroleum Hydrocarbon (TPH) contaminated soils. TPH was treated within the standard of concern, and E. coli was analyzed to be high in organic matter, and the fertilizer specification was satisfied by applying the optimum composting conditions at 70℃, but the organic matter content was lower than the fertilizer specification. As a result of continuous washing test, Cd has 5 levels of residual material in fine soil. Cu and Zn were mostly composed of ion exchange properties (stage 1), carbonates (stage 2), and iron / manganese oxides (stage 3), which facilitate easy separation of contamination. As a result of applying acid dissolution and multi-stage washing step by step, hydrochloric acid, 1.0M, 1: 3, 200rpm, 60min was analyzed as the optimal washing factor. Most of the contaminated sediments were found to satisfy the Soil Environmental Conservation Act's standards. Therefore, as a result of the applicability test of this study, soil with high heavy metal contamination was used as aggregate by applying soil cleaning after pre-treatment. It was possible to verify that it was efficient to use organic and oil-contaminated soil as compost Maturity after exterminating contaminants and E. coli by applying composting.