• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10407-10412
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• 2015

Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.12
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• pp.4773-4780
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• 2014

Background: To investigate the effects of small interference RNA (siRNA) targeting BCR/ABL mRNA on proliferation and apoptosis in the K562 human chronic myeloid leukemia (CML) cell line and to provide a theoretical rationale and experimental evidence for its potential clinical application for anti-CML treatment. Materials and Methods: The gene sequence for BCR/ABL mRNA was found from the GeneBank. The target gene site on the BCR/ABL mRNA were selected according to Max-Planck-Institute (MPI) and rational siRNA design rules, the secondary structure of the candidate targeted mRNA was predicted, the relevant thermodynamic parameters were analyzed, and the targeted gene sequences were compared with BLAST to eliminate any sequences with significant homology. Inhibition of proliferation was evaluated by MTT assay and colony-formation inhibiting test. Apoptosis was determined by flow cytometry (FCM) and the morphology of apoptotic cells was identified by Giemsa-Wright staining. Western blotting was used to analyze the expression of BCR/ABL fusion protein in K562 cells after siRNA treatment. Results: The mRNA local secondary structure calculated by RNA structure software, and the optimal design of specific siRNA were contributed by bioinformatics rules. Five sequences of BCR/ABL siRNAs were designed and synthesized in vitro. Three sequences, siRNA1384, siRNA1276 and siRNA1786, which showed the most effective inhibition of K562 cell growth, were identified among the five candidate siRNAs, with a cell proliferative inhibitory rate nearly 50% after exposure to 12.5nmol/L~50nmol/L siRNA1384 for 24,48 and 72 hours. The 50% inhibitory concentrations ($IC_{50}$) of siRNA1384, siRNA1276 and siRNA1786 for 24hours were 46.6 nmol/L, 59.3 nmol/L and 62.6 nmol/L, respectively, and 65.668 nmol/L, 76.6 nmol/L, 74.4 nmol/L for 72 hours. The colony-formation inhibiting test also indicated that, compared with control, cell growth of siRNA treated group was inhibited. FCM results showed that the rate of cell apoptosis increased 24 hours after transfecting siRNA. The results of annexinV/PI staining indicated that the rate of apoptosis imcreased (1.53%, 15.3%, 64.5%, 57.5% and 21.5%) following treamtne with siRNAs (siRNA34, siRNA372, siRNA1384, siRNA1276 and siRNA1786). Morphological analysis showed td typical morphologic changes of apoptosis such as shrunken, fragmentation nucleus as well as "apoptotic bodies" after K562 cell exposure to siRNA. Western blot analysis showed that BCR/ABL protein was reduced sharply after a single dose of 50nmol/L siRNA transfection. Conclusions: Proliferation of K562 cells was remarkbly inhibited by siRNAs (siRNA1384, siRNA1276 and siRNA1786) in a concentration-dependent manner in vitro, with effective induction of apoptosis at a concentration of 50 nmol/L. One anti-leukemia mechanism in K562 cells appeared that BCR/ABL targeted protein was highly down-regulated. The siRNAs (siRNA1384, siRNA1276 and siRNA1786) may prove valuable in the treatment of CML.

• Journal of Korean Society of Environmental Engineers
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• v.31 no.11
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• pp.1007-1018
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• 2009

Hydrocyclone is widely used in industry, because of its simplicity in design, high capacity, low maintenance and operational cost. The separation action of a hydrocyclone treating particulate slurry is a consequence of the swirling flow that produces a centrifugal force on the fluid and suspended particles. In spite of hydrocyclone have many advantage, the application for treatment of urban stormwater case study were rare. We conducted a laboratory scale study on treatable potential of micro particles using hydrocyclone filter (HCF) that was a combined modified hydrocyclone with perlite filter cartridge. Since it was not easy to use actual storm water in the scaled-down hydraulic model investigations, it was necessary to reproduce ranges of particles sizes with synthetic materials. The synthesized storm runoff was made with water and addition of particles; ion exchange resin, road sediment, commercial area manhole sediment, and silica gel particles. Experimental studies have been carried out about the particle separation performance of HCF-open system and HCF-closed system. The principal structural differences of these HCFs are underflow zone structure and vortex finder. HCF was made of acryl resin with 120 mm of diameter hydrocyclone and 250 mm of diameter filter chamber and overall height of 800 mm. To determine the removal efficiency for various influent concentrations of suspended solids (SS) and chemical oxygen demand (COD), tests were performed with different operational conditions. The operated maximum of surface loading rate was about 700 $m^3/m^2$/day for HCF-open system, and 1,200 $m^3/m^2$/day for HCF-closed system. It was found that particle removal efficiency for the HCF-closed system is better than the HCF-open system under same surface loading rate. Results showed that SS removal efficiency with the HCF-closed system improved by about 8~20% compared with HCF-open system. The average removal efficiency difference for HCF-closed system between measurement and CFD particle tracking simulation was about 4%.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.1
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• pp.23-32
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• 2010

The capacity of natural purification was limited by the interruption of natural flow and the problems such as eutrophication were occurred by nutritive salts accumulation in stagnant stream. Moreover, the inflow of non-point sources causes non-degradable materials to increase in stagnant stream. In this study, an upflow biological activated carbon (BAC) biofilm process comprised of anoxic, aerobic 1, and aerobic 2 reactors were introduced for treatment of stagnant stream and SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$, TN, and TP were monitored in the upflow BAC biofilm reactors with continuous cycling. In order to simulate stagnant stream, the lake water of amusement park and golf course were stored as influent in a tank of $2m^3$ and hydraulic retention time (HRT) was changed into 6, 4, and 2 hours. At HRT 4hr and the lake water of amusement park as influent, the removal efficiencies of SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$, TN, and TP showed the best water quality improvement and were 69.8, 83.0, 91.3, 74.1, 74.7, and 88.9%, respectively. At HRT 4hr and the lake water of golf course as influent, the removal efficiencies of SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$, TN and TP were 78.5, 78.0, 80.2, 74.9, 55.6 and 97.5%, respectively. As the results of polymerase chain reaction - denaturing gel gradient electrophoresis (PCR-DGGE), microbial community was different depending on influent type. Fluorescence in situ hybridization (FISH) showed that nitrifying bacteria was dominant at HRT 4 hr. The biomass amount and microbial activities by INT-DHA test were not decrease even at lower HRT condition. In this study, the upflow BAC biofilm process would be considered to the water quality improvement of stagnant stream.

• The Journal of the Petrological Society of Korea
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• v.9 no.4
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• pp.238-253
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• 2000

In the eastern part of the Euiseong Basin acidic~intermediate volcanic rocks widely distribute on the Cretaceous sedimentary basement. Coeval granitic rocks and dyke rocks intruded into the volcanic rocks. Volcanic stratigraphy of study area are andesite lava, dacitic lapilli tuff, dacitic flow-banded lava, rhyolitic bedded tuff, rhyolitic massive tuff, dacitic massive lava, rhyolitlc welded tuff occur from the lower to the upper strata. $SiO_2$ content of the volcanic rocks range from 51 to 74 wt.%. With the increase of $SiO_2$, the contents of $TiO_2$, $Al_2$$O_3$, MgO, FeOT MnO, CaO, $P_2$$O_{5}$ decrease but those of $K_2$O increase. The contents of $Na_2$O show dispersive variation. This trend is quite sim-ilar to the major oxide variation in the volcanic rocks from the Yucheon sub-basin. The geochemical natures indicate that the volcanic rocks in the study area are discriminated to the island-arc type high K to medium K calc-alkaline rocks. The compositional variation of the volcanic rocks can be explained by the plagioclase fractionation of the volcanic magmas originated from similar source materials. The volcanic stratigraphy seems to have formed by at least two eruptive sequences of andesitic to rhyolitic and dacitic to rhyolitic magmas which underwent crystallization differentiation.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.4
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• pp.299-305
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• 2010

Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.

• Tuberculosis and Respiratory Diseases
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• v.52 no.3
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• pp.251-259
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• 2002

Background : Reduced lung compliance and increased lung resistance are the primary lung mechanical abnormalities in acute respiratory distress syndrome (ARDS). Although there is little information regarding the mechanisms responsible for the increases in the respiratory resistance of ARDS, bronchodilators have been frequently administered in mechanically ventilated ARDS patients. To determine the effect of a bronchodilator on the respiratory mechanics depending on the level of applied positive end-expiratory pressure (PEEP), the changes in the respiratory mechanics by salbutamol inhalation was measured under the variable PEEP level in patients with ARDS. Materials and Methods : Fifteen mechanically ventilated paralyzed ARDS patients (14 of male, mean age 57 years) were enrolled in this study. The respiratory system compliance, and the maximum and minimum inspiratory resistance were obtained by the end-inspiratory occlusion method during constant flow inflation using the CP-100 pulmonary monitor (Bicore, Irvine, CA, USA). The measurements were performed at randomly applied 8, 10 and 12 cm $H_2O$ PEEP before and 30 mins after administrating salbutamol using a meter-dose-inhaler (100ug${\times}$6). Results : 1) The maximum inspiratory resistance of the lung was higher than the reported normal values due to an increase in the minimal inspiratory resistance & additional resistance. 2) The maximum inspiratory resistance and peak airway pressure were significantly higher at 12cm $H_2O$ of PEEP compared with those at 10cm $H_2O$ of PEEP. 3) Salbutamol induced a significant decrease in the maximum and the minimum inspiratory resistance but no significant change in the additional resistance only was observed at 12cm $H_2O$ of PEEP(from $15.66{\pm}1.99$ to $13.54{\pm}2.41$, from $10.24{\pm}2.98$ to $8.04{\pm}2.34$, and from $5.42{\pm}3.41$ to $5.50{\pm}3.58cm$ $H_2O$/L/sec, respectively). 4)The lung compliance did not change at the applied PEEP and salbutamol inhalation levels. Conclusion : The bronchodilator response would be different depending on the level of applied PEEP despite the increased respiratory resistance in patients with ARDS.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.1
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• pp.61-67
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• 2016

An HPLC analysis method was developed for standard determinations of chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid as functional health materials in Ligularia fischeri extract. HPLC was performed on a $C_{18}$ Kromasil column ($4.6{\times}250mm$, $5{\mu}m$ column) with a gradient elution of 0.1% (v/v) trifluoroacetic acid and acetonitrile at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analytes were detected at 330 nm. The HPLC method was validated in accordance with the International Conference on Harmonization guideline of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation for the four compounds were 3.0~14.6 and $9.2{\sim}44.4{\mu}g/mL$, respectively. Calibration curves showed good linearity ($r^2$ > 0.999), and the precision of analysis was satisfied (less than 0.9%). Recoveries of quantified compounds ranged from 98.96 to 101.81%. This result indicates that the established HPLC method is very useful for the determination of marker compounds in Ligularia fischeri leaf extracts.

• The Korean Journal of Nuclear Medicine
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• v.37 no.2
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• pp.94-102
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• 2003

Purpose : Cerebral blood flow (CBF) reactivity to acetazolamide (ACZ) is useful to select patients with hemodynamic failure. However, it is still a matter of speculation that varying degrees of regional CBF increases after ACZ administration represent the severity or stage of regional hemodynamic failure as assessed by positron emission tomography (PET). We studied to elucidate whether ACZ challenge $^{123}I-IMP$ brain single photon emission tomography (SPECT) can accurately grade the seventy of regional hemodynamic failure. Materials and Methods: Eighteen patients (M: 16, F: 2, average age: 61 years) with unilateral occlusive disease of the internal carotid artery or the trunk of the middle cerebral artery (MCA). Patients undewent $^{123}I-IMP$ brain SPECT study with acetazolamide challenge and PET study was carried out within 2 weeks before and after SPECT study. Five healthy volunteers with a mean age of 48 years (range: 28-73 yr, M: 3, F: 2) underwent PET studies to determine normal values. In SPECT study, an asymmetry index (Al)-the percentage of radioactivity of region of interest (ROI) in the occlusive cerebrovascular lesion to the contralateral homologous ROI-was used for numerical evaluation of relative $^{123}I-IMP$ distribution. In PET study, regional CBF, oxygen extraction fraction (OEF), cerebral metabolic rate of oxygen ($CMRO_2$) and cerebral blood volume (CBV) values were measured with $^{15}O-labeled$ gas inhalation method and the values were used for comparison with Al (Al during acetazolamide challenge-Al of basal study) on the SPECT study. ROls were classified by severity into three groups (normal, stage I and stage II). Results: Mean values of Al in areas with normal, stage I and stage II hemodynamic failure were $6.25{\pm}7.77%\;(n=107),\;-10.38{\pm}10.41%\:(n=117)\;and\;13.30{\pm}10.51%\;(n=140)$, respectively. Al significantly differed with each groups (p<0.05). Correlation between Al and CBF, OEF and CBV/CBF in hemisphere with occlusive cerebrovascular lesion was 0.20 (p<0.01), -0.28 (p<0.01) and -0.28 (p<0.01), respectively. Conclusion: We concluded that $^{123}I-IMP$ brain SPECT with acetazolamide challenge could determine the severity ad stage of regional hemodynamic failure as assessed by PET.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.18
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• pp.7849-7855
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• 2014

Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.