• Journal of Dental Anesthesia and Pain Medicine
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• v.16 no.4
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• pp.263-271
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• 2016

Background: Bone injury is common in many clinical situations, such as surgery or trauma. During surgery, excessive reactive oxygen species (ROS) production decreases the quality and quantity of osteoblasts. Remifentanil decreases ROS production, reducing oxidative stress and the inflammatory response. We investigated remifentanil's protective effects against $H_2O_2$-induced oxidative stress in osteoblasts. Methods: To investigate the effect of remifentanil on human fetal osteoblast (hFOB) cells, the cells were incubated with 1 ng/ml of remifentanil for 2 h before exposure to $H_2O_2$. For induction of oxidative stress, hFOB cells were then treated with $200{\mu}M$ $H_2O_2$ for 2 h. To evaluate the effect on autophagy, a separate group of cells were incubated with 1 mM 3-methyladenine (3-MA) before treatment with remifentanil and $H_2O_2$. Cell viability and apoptotic cell death were determined via MTT assay and Hoechst staining, respectively. Mineralized matrix formation was visualized using alizarin red S staining. Western blot analysis was used to determine the expression levels of bone-related genes. Results: Cell viability and mineralized matrix formation increased on remifentanil pretreatment before exposure to $H_2O_2$-induced oxidative stress. As determined via western blot analysis, remifentanil pretreatment increased the expression of bone-related genes (Col I, BMP-2, osterix, and $TGF-{\beta}$). However, pretreatment with 3-MA before exposure to remifentanil and $H_2O_2$ inhibited remifentanil's protective effects on hFOB cells during oxidative stress. Conclusions: We showed that remifentanil prevents oxidative damage in hFOB cells via a mechanism that may be highly related to autophagy. Further clinical studies are required to investigate its potential as a therapeutic agent.

• Development and Reproduction
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• v.12 no.3
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• pp.221-229
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• 2008

For the clinical application, it is needed to keep characteristics of stem cells after storage for a long time. In the present study, we examined stem cell properties of human cord-derived stem cells (HUC) after cryopreservation. Cells were isolated from human umbilical cord and cultured in vitro. At passage 2 or 3, HUC were suspended at a concentration of $1.0{\times}10^6/m{\ell}$ in cryomedium consisting of DMSO and FBS. After freezing at $-80^{\circ}C$ overnight, HUC were cryopreserved at $-196^{\circ}C$ nitrogen gas. After 6 months, HUC were thawed and cultured in vitro. Assessment for the stem cell properties was made upon the morphology, population doubling time, and expression profiles of genes and various proteins. Cryopreserved HUC showed more than 70% viability and maintained fibroblast-like morphology similar to HUC before cryopreservation. Throughout the culture, they underwent average 42.8 doublings and produced $6.75{\times}{10^{18}}$ cells. RT-PCR analyses showed that cryopreserved HUC expressed Oct-4, nanog, SCF, NCAM, nestin, GATA-4, BMP4, and HLA-1 genes. They did not express Brachyury and HLA-DR genes. Immunocytochemical studies showed that cryopreserved HUC reacted with antibodies against SSEA-3, -4, Thy-1, vimentin, fibronectin, HCAM, ICAM, HLA-1 proteins. They did not react with antibody against HLA-DR protein. Theses genes and proteins expression patterns of cryopresserved HUC were similar to those of HUC before cryopreservation. These results suggest that cryopreserved HUC could retain proliferative potential and they expressed various genes and proteins similar to HUC before cryopreservation. Thus, cryopreservation might be useful for HUC for future research and clinical application.

• Journal of environmental and Sanitary engineering
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• v.20 no.1 s.55
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• pp.64-75
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• 2005

The total production of food waste was about 11,398ton/day('03) in Korea. Also, food waste was treated by landfill, incineration, reuse and anaerobic digestion. The method of food waste treatment depended primarily on landfill. However, the method of landfill causing social problems was prevented to treat food waste in the first of January 2005.12) Thus, anaerobic digestion is an important method to treat food waste because of possibility of energy recovery as methane gas. In this study, the possibility of food waste treatment containing high organic material and low pH in the one stage anaerobic reactor to save cost and time and energy recovery using $CH_{4}$ gas by the hybrid anaerobic reactor (HAR) was measured. The HAR was designed by combing the merits of the anaerobic filter (AF) to minimize the microorganism shock when food waste of very low pH was injected and up-flow anaerobic sludge blanket (UASB) to prevent from plugging and channeling phenomena by large suspended solids when semi solids were injected. Granule was packed in the section of HAR. The purpose of the BMP experiment was to measure the amount of methane generated when organic material was resolved under anaerobic conditions, to grasp bio resolution of organic material. Total accumulated methane production per VS amount was $0.471(m^{3}/\cal{kg}\;VS)$. So, the value was about $81.2\%$ of theoretical methane production which was $0.58(m^{3}/\cal{kg}\;VS)$ by elementary analysis and organic matter removal velocity (K) was $0.18(d^{-1})$. From these results, food waste was treated by anaerobic treatment. From this study, $CH_{4}$ generation from food waste (11,398 ton/day) could be estimated. By using an energy conversion factor of Braun's study, $5.97KWh/m^{3}\;CH4,\;60\%\;of\;CH_{4}$ gas generation, the amount of total energy producing food waste is to 6,727MWh/day. It could be confirmed that energy recovery using $CH_{4}$ gas was possible. Above these results, food waste containing organic matters of high concentration could be treated in HRT 30 days under an anaerobic condition, using the hybrid anaerobic reactor and reuse of $CH_{4}$ gas was possible.

• International Journal of Highway Engineering
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• v.9 no.4
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• pp.185-192
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• 2007

Newly constructed road is a requisite to be able to carry out BMPs (Best Management Practices) under TMDL(Total Maximum Daily Load) program of the Ministry of Environment. BMPs require pollutant source control during road construction and wash off reduction plan as well as maintenance practices subsequent to construction on the purpose of discharging the minimum wash off non-point source pollutants. The objective of this study is to provide supportive discharged data in evaluating the discharged non-point pollutant load from a highway toll gate area. It can be applied to manage non-point source pollutants on roads. The results validate the first flush phenomenon that it is known to be one of the wash off characteristics in paved area. In addition, the load per unit area and load per unit rainfall duration applying EMC are calculated. The mean load per unit rainfall duration is assessed to be $533.7mg/m^2-hr$ for TSS, $396.2mg/m^2-hr$ for COD, $17.0mg/m^2-hr$ for TN, and $4.8mg/m^2-hr$ for TP. These results show the unitload taken from monitoring are higher than the unit load suggested in the TMDL. It is important to adopt real pollutant unit for road to be able to perform BMP successfully.

• Journal of Korean Medicine Rehabilitation
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• v.28 no.1
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• pp.1-17
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• 2018

Objectives The purpose of this study was to evaluate the effect of Jeopgolsan (JGS) extract on anti-oxidant, anti-inflammatory activities in RAW 264.7 cells and on factors related with fracture healing in skull fractured rat. Methods Experimental animals were divided into four groups: normal group without any treatment (Normal), contral group were treated orally with distilled water (Control), Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 200) and Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 400). Rats in each group except the normal group were induced fractures in the skull. The 1,1-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity were measured to evaluate antioxidant activity. The production of nitric oxide (NO), $interleukin-1{\beta}$ ($IL-1{\beta}$), interleukin-6 (IL-6) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) in the RAW 264.7 cells were measured to evaluate anti-inflammatory activity. The production of osteocalcin calcitonin, carboxy-terminal telepeptides of type II collagen (CTX II), transforming growth $factor-{\beta}$ ($TGF-{\beta}$), bone morphogenetic protein-2 (BMP-2), Insulin and alkaline phosphatase (ALP) in serum of rats were measured to evaluate the effects of fracture healing at 0, 2, 4, and 6th week. X-rays were taken every 3 week from 0 to 6th week to evaluate fracture healing effect. Results 1. No cytotoxicity was observed. 2. DPPH and ABTS radical scavenging activity were increased in a concentration dependent manner, indicating anti-oxidant effect. 3. NO, $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ were not significantly changed, indicating no anti-inflammatory effect. 4. Osteocalcin, Calcitonin, $TGF-{\beta}$ and ALP were significantly increased in the experimental groups. 5. CTX II, insulin were significantly decreased in the expermental groups. 6. Radiologic examination showed that union of fracture was promoted. Conclusions From above results, JGS showed significant results in factors related with fracture healing and radiologic examination. Threfore, JGS is expected to be effective in the treatment of fracture.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.1
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• pp.23-34
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• 2009

Objectives: Mesenchymal stem cells (MSC) comprise a promising tool for cellular therapy. It is known that long-term in vitro culture of human bone marrow and adipose tissue derived-MSCs lead to a reduction of life span and a change of stem-like characters. The aim of our study was to examine whether stem cell properties of human umbilical cord-derived stem cells (HUC) could be affected by in vitro expansion. Methods: HUC were isolated from human umbilical cord and cultured for 10 passages in vitro. Morphology and population doubling time (PDT) were investigated, and changes of stem cell properties were examined using RT-PCR and immunocytochemistry during serial subcultures. Results: Morphology and PDT of HUC began to change slightly from the 7th passage (p7). Expression level of nestin and vimentin mRNAs increased along with the culture period from p4 until p10. In contrast, expression level of SCF mRNA decreased during the same culture period. Expression level of Oct-4 and HNF-4${\alpha}$ mRNAs was not significantly changed throughout the culture period until p10. Expression level of BMP-4, FGF-5, NCAM and HLA-ABC mRNAs appeared to increase as the culture continued, however, the difference was not significant. Immunocytochemical studies showed that HUC at p3, p6 and p9 positively were stained with antibodies against SSEA-3 and SSEA-4 proteins. Interestingly, staining intensity of HUC for ICAM-1 and HLA-ABC gradually increased throughout the culture period. Intensity against thy-1 and fibronectin antibodies increased at p9 while that against TRA-1-60 and VCAM-1 antibodies began to decrease at p6 until p9. Conclusions: These results suggest that HUC change some of their stem cell characteristics during in vitro culture. Development of culture system might be needed for the maintenance of characteristics.

• Journal of Applied Biological Chemistry
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• v.57 no.1
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• pp.95-101
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• 2014

Nonpoint pollution sources from agricultural activities are a major cause of water quality impairment. A nutrient management program utilizes farm practices that maintain efficient crop production systems and control agricultural nonpoint pollution sources. The objectives of present study were to identify appropriate best management practices (BMPs) according to changes of cropping system of main upland crop for reducing AGNPs loadings and to simulate the effects of the application of the several BMPs scenarios in Saemangeum watershed. The selected BMP scenarios were: 1) to convert naked barley and hulled barley to hairy vetch or chinese milk vetch, 2) to convert red pepper to soybean crop, and 3) to combine two scenarios, converting naked barley and hulled barley to hairy vetch or chinese milk vetch + converting red pepper to soybean crop. As a result of BMPs application, the crop requirement of nitrogen and phosphorus for upland crop reduced nitrogen by 41% and phosphorus by 47% in scenario 1, whereas scenario 2 reduced nitrogen by 30% and phosphorus by 23%. Overall, scenario 3 reduced nitrogen by 72% and phosphorus by 70% in agricultural non-point pollution sources associated with chemical fertilizer and livestock manure in Saemangeum watershed.

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.269-279
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• 2004

백악질 세포의 분리 및 배양방법을 확립하고, 이를 이용하여 백악질 세포의 형질특성을 알아보고자 하였다. 교정목적으로 발거된 소구치를 이용하여, 치은섬유아세포, 치주인대 세포 및 백악질 유래세포를 분리, 배양하였다. 백악질 유래 세포 배양시에는 백악질을 절제한 후 Collagenase P를 이용하여 백악질 유래 세포 외의 다른 세포의 개제를 배제하였고, 기질을 분해하여 세포의 분리 및 배양이 용이하도록 하였다. 분리 및 배양시기의 세포의 형태를 광화현미정을 이용하여 관찰하였다. 조골세포의 특성을 가지는 SaOs-2 세포를 대조군으로 이용하여 분리 및 배양된 세포군들을 동일한 조건으로 배양하였다. 3일 및 7일째에 세포증식도를 측정하였고 7일째에 ALPase 효소 활성도를 측정하였다. 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 osteopontin(OPN), Alkaline phosphatase(ALP), type I collagen(COL-I), Bone sialoprotein(BSP), BMP-2 및 osteocalcin(OC)의 발현을 비교 관찰하였다. 백악질 유래 세포의 분리 및 배양을 시도한 5명의 치아 중에서 3명의 치아에서 세포군을 배양해 낼 수 있었다. 배양한 백악질 유래 세포는 섬유아세포와 유사한 형태와 증식을 보였다. ALPase 효소 활성도 검사 결과 백악질 유래 세포는 SaOs-2 세포보다 낮은 활성도를 나타내었으며, 배양된 세포의 RT-PCR 결과 백악질 유래 세포군에서는 ALPase의 발현이 나타나지 않았고, 다른 조골세포 표식자의 발현도 낮게 나타났다. 이는 백악질 유래 세포가 조골세포 및 다른 대조군의 세포와는 다른 형질 특성을 가지고 있다는 것을 시사한다. 이상의 관찰결과로 사람의 백악질 유래 세포롤 백악질의 절제 및 효소처리 방법으로 효과적인 분리 및 배양이 가능하며, 이는 향후 백악질 세포의 형질 특성 및 백악질 형성의 분자적 기전을 파악하는 중요한 연구자료로 활용 될 수 있을 것으로 사료된다.

• Journal of Physiology & Pathology in Korean Medicine
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• v.31 no.6
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• pp.374-379
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• 2017

Hair loss affects interpersonal relationships and causes psychological stress. In this study, we investigated the antioxidant activity of Cynanchi Wilfordii Radix (CWR) and its effects on dermal papilla (DP) cells. Antioxidant efficacy was examined by ABTS assay. To confirm the effect on cell activity, MTS assay was performed and cell count was directly measured by hemocytometer. The mRNA expression of genes involved in hair formation and hair loss formation was confirmed by quantitative RT-PCR. CWR has a strong antioxidant activity. Cell viability of DP cells was increased to 118.5% by treatment of 0.5 mg/ml CWR for 24 hours, but the effect on the cell number was insignificant. These results suggest that CWR increases mitochondrial activity without promoting cell proliferation. Treatment of DP cells with 0.5 mg/ml CWR resulted in 48.5% reduction of mRNA expression of type 2 $5{\alpha}$-reductase, a major cause of male hair loss. In addition, mRNA expression of bone morphogenetic pretein (BMP), fibroblast growth factor (FGF)7, and FGF10, which are closely related to hair growth, was also decreased. Reactive oxygen species (ROS) acts as a cause of hair loss. The excellent antioxidant efficacy of CWR is thought to be able to effectively remove ROS. The dihydrotestosterone produced by type 2 $5{\alpha}$-reductase in DP cells is a potent inducer of male pattern hair loss. The inhibitory effect of type 2 $5{\alpha}$-reductase mRNA on DP cells induced by CWR may induce a positive therapeutic effect of male pattern hair loss.

• 한국신재생에너지학회:학술대회논문집
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• 2010.11a
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• pp.101-101
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• 2010

Anaerobic digestion(AD) is the most promising method for treating and recycling of different organic wastes, such as organic fraction of municipal solid waste, household wastes, animal manure, agro-industrial wastes, industrial organic wastes and sewage sludge. During AD, i.e. organic materials are decomposed by anaerobic forming bacteria and fina1ly converted to excellent fertilizer and biogas which is a mixture of carbon dioxide and methane. AD has been one of the leading technologies that can make a large contribution to produce renewable energy and to reduce $CO_2$ and other green-house gas(GHG) emission, it is becoming a key method for both waste treatment and recovery of a renewable fuel and other valuable co-products. Currently some 80% of the world's overall energy supply of about 400 EJ per year in derived from fossil fuels. Nevertheless roughly 10~15% of this demand is covered by biomass resources, making biomass by far the most important renewable energy source used to date. The representative biofuels produced from the biomass are bioethanol, biodiesel and biogas, and currently biogas plays a smaller than other biofuels but steadily growing role. Traditionally anaerobic digestion applied for different biowaste e.g. sewage sludge, manure, other organic wastes treatment and stabilization, biogas has become a well established energy resource. However, the biowaste are fairly limited in respect to the production and utilization as renewable source, but the plant biomass, the so called "energy crops" are used for more biogas production in EU countries and the investigation on the biomethane potential of different crops and plant materials have been carried out. In Korea, with steadily increasing oil prices and improved environmental regulations, since 2005 anaerobic digestion was again stimulated, especially on the biogasification of different biowastes and agro-industrial biomass including "energy crops". This study have been carried out to investigate anaerobic biodegradability by the biochemical methane potential(BMP) test of animal manures, different forage crops i.e. "energy crops", plant and industrial organic wastes in the condition of thermophilic temperature, The biodegradability of animal manure were 63.2% and 58.2% with $315m^3CH_4/tonVS$ of cattle slurry and $370m^3CH_4/tonVS$ of pig slurry in ultimate methane yields. Those of winter forage crops were the range 75% to 87% with ultimate methane yield of $378m^3CH_4/tonVS$ to $450m^3CH_4/tonVS$ and those of summer forage crops were the range 81% to 85% with ultimate methane yield of $392m^3CH_4/tonVS$ to $415m^3CH_4/tonVS$. The forge crops as "energy crops" could be used as good renewable energy source to increase methane production and to improve biodegradability in co-digestion with animal manure or only energy crop digestion.