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Establishment of Paclitaxel-resistant Breast Cancer Cell Line and Nude Mice Models, and Underlying Multidrug Resistance Mechanisms in Vitro and in Vivo

  • Chen, Si-Ying;Hu, Sa-Sa;Dong, Qian;Cai, Jiang-Xia;Zhang, Wei-Peng;Sun, Jin-Yao;Wang, Tao-Tao;Xie, Jiao;He, Hai-Rong;Xing, Jian-Feng;Lu, Jun;Dong, Ya-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6135-6140
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    • 2013
  • Background: Breast cancer is a common malignant tumor which affects health of women and multidrug resistance (MDR) is one of the main factors leading to failure of chemotherapy. This study was conducted to establish paclitaxel-resistant breast cancer cell line and nude mice models to explore underlying mechanisms of MDR. Methods: The breast cancer drug-sensitive cell line MCF-7 (MCF-7/S) was exposed in stepwise escalating paclitaxel (TAX) to induce a resistant cell line MCF-7/TAX. Cell sensitivity to drugs and growth curves were measured by MTT assay. Changes of cell morphology and ultrastructure were examined by optical and electron microscopy. The cell cycle distribution was determined by flow cytometry. Furthermore, expression of proteins related to breast cancer occurrence and MDR was tested by immunocytochemistry. In Vivo, nude mice were injected with MCF-7/S and MCF-7/TAX cells and weights and tumor sizes were observed after paclitaxel treatment. In addition, proteins involved breast cancer and MDR were detected by immunohistochemistry. Results: Compared to MCF-7/S, MCF-7/TAX cells had a higher resistance to paclitaxel, cross-resistance and prolonged doubling time. Moreover, MCF-7/TAX showed obvious alterations of ultrastructure. Estrogen receptor (ER) expression was low in drug resistant cells and tumors while expression of human epidermal growth factor receptor 2 (HER2) and Ki-67 was up-regulated. P-glycoprotein (P-gp), lung resistance-related protein (LRP) and glutathione-S-transferase-${\pi}$ (GST-${\pi}$) involved in the MDR phenotype of resistant cells and tumors were all overexpressed. Conclusion: The underlying MDR mechanism of breast cancer may involve increased expression of P-gp, LRP and GST-${\pi}$.

TATA box binding protein and ribosomal protein 4 are suitable reference genes for normalization during quantitative polymerase chain reaction study in bovine mesenchymal stem cells

  • Jang, Si-Jung;Jeon, Ryoung-Hoon;Kim, Hwan-Deuk;Hwang, Jong-Chan;Lee, Hyeon-Jeong;Bae, Seul-Gi;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.12
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    • pp.2021-2030
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    • 2020
  • Objective: Quantitative polymerase chain reaction (qPCR) has been extensively used in the field of mesenchymal stem cell (MSC) research to elucidate their characteristics and clinical potential by normalization of target genes against reference genes (RGs), which are believed to be stably expressed irrespective of various experimental conditions. However, the expression of RGs is also variable depending on the experimental conditions, which may lead to false or contradictory conclusions upon normalization. Due to the current lack of information for a clear list of stable RGs in bovine MSCs, we conducted this study to identify suitable RGs in bovine MSCs. Methods: The cycle threshold values of ten traditionally used RGs (18S ribosomal RNA [18S], beta-2-microglobulin [B2M], H2A histone family, member Z [H2A], peptidylprolyl isomerase A [PPIA], ribosomal protein 4 [RPL4], succinate dehydrogenase complex, subunit A [SDHA], beta actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box binding protein [TBP], and hypoxanthine phosphoribosyltrasnfrase1 [HPRT1]) in bovine bone marrow-derived MSCs (bBMMSCs) were validated for their stabilities using three types of RG evaluation algorithms (geNorm, Normfinder, and Bestkeeper). The effect of validated RGs was then verified by normalization of lineage-specific genes (fatty acid binding protein 4 [FABP4] and osteonectin [ON]) expressions during differentiations of bBMMSCs or POU class 5 homeobox 1 (OCT4) expression between bBMMSCs and dermal skins. Results: Based on the results obtained for the three most stable RGs from geNorm (TBP, RPL4, and H2A), Normfinder (TBP, RPL4, and SDHA), and Bestkeeper (TBP, RPL4, and SDHA), it was comprehensively determined that TBP and RPL4 were the most stable RGs in bBMMSCs. However, traditional RGs were suggested to be the least stable (18S) or moderately stable (GAPDH and ACTB) in bBMMSCs. Normalization of FABP4 or ON against TBP, RPL4, and 18S presented significant differences during differentiation of bBMMSCs. However, although significantly low expression of OCT4 was detected in dermal skins compared to that in bBMMSCs when TBP and RPL4 were used in normalization, normalization against 18S exhibited no significance. Conclusion: This study proposes that TBP and RPL4 were suitable as stable RGs for qPCR study in bovine MSCs.

A Study on Characteristics of an Integrated Urea-SCR Catalytic Filter System for Simultaneous Reduction of Soot and NOX Emissions in ECU Common-rail Diesel Engines (ECU 커먼레일 디젤기관에 있어서 매연 및 NOX 배출물 동시 저감용 일체형 요소-SCR 촉매필터 시스템의 특성에 관한 연구)

  • Bae, Myung-Whan
    • Transactions of the Korean Society of Automotive Engineers
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    • v.22 no.4
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    • pp.111-120
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    • 2014
  • The aim of this study is to develop an integrated urea-SCR catalytic filter system for reducing soot and $NO_X$ emissions simultaneously in diesel engines. In this study, the characteristics of exhaust emissions relative to reactive activation temperature under four kinds of engine loads are experimentally investigated by using a four-cycle, four-cylinder, direct injection type, water-cooled turbo intercooler ECU common-rail diesel engine with the integrated urea-SCR $MnO_2-V_2O_5-WO_3/TiO_2/SiC$ catalytic filter system operating at three kinds of engine speeds. The urea-SCR reactor is used to reduce $NO_X$ emissions, and the catalytic filter system is used to reduce soot emissions. The reactive activation temperature is very important for reacting a reducing agent with exhaust emissions. The reactive activation temperatures in this experiment is applied to 523, 573 and 623 K. The fuel is sprayed by the pilot and main injections at the variable injection timing between BTDC $15^{\circ}$ and ATDC $1^{\circ}$ according to experimental conditions. It is found that the $NO_X$ conversion rate is the highest as 83.9% at the reactive activation temperature of 523 K in all experimental conditions of engine speed and load, and the soot emissions shown by the average reduction rate of approximately 93.3% are almost decreased below 0.6% in all experimental conditions regardless of reactive activation temperatures. Also, the THC and CO emissions by oxidation reaction of Mn, V and Ti are shown in the average reduction rates of 70.3% and 38% regardless of all experimental conditions.

Influence of the Monitoring Interval and Intake Pattern for the Evaluation of Intake (내부피폭 감시주기 및 섭취형태가 방사성핵종 섭취량 평가에 미치는 영향)

  • Jong-Il Lee;Tae-Young Lee;Si-Young Chang;Jai-Ki Lee
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.2 no.1
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    • pp.53-59
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    • 2004
  • A variety of factors such as the pattern of intake (acute or chronic), monitoring interval and the characteristics of the radionuclides could have a significant influence on the estimates for the intake and internal dose. The relative differences of the assessed intakes based on the assumption of an acute intake to that of a chronic intake were evaluated by using the predicted bioassay quantity in the whole body or organs for an acute and chronic intake through the inhalation of $^{125}$ I, $^{137}$ C, $^{235}$ U with the AMAD of 1 ${\mu}{\textrm}{m}$ and 5 ${\mu}{\textrm}{m}$ for the monitoring intervals of 7, 14, 30, 60, 90, 120, 180, 360 days, respectively, The relative difference of the assessed intakes based on the intake pattern is affected by the monitoring interval, radionuclide and absorption type, but the particle size has little influence on the difference of the assessed intakes based on the intake pattern. The maximum monitoring interval, which is defined as the monitoring interval that the relative difference of the assessed intakes based on the assumption of an acute intake to that of a chronic intake is less than 10%, is 60 d for $^{125}$ I with Type F, 180 d for $^{137}$ C with Type F, 90 d for $^{235}$ U with Type M, and 360 d for $^{235}$ U with Type S. It was concluded that an intake pattern has little influence on the estimates of the assessed intake in the case where the monitoring interval is shorter than the maximum monitoring interval for each radionuclide.

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HIPIMS Arc-Free Reactive Deposition of Non-conductive Films Using the Applied Material ENDURA 200 mm Cluster Tool

  • Chistyakov, Roman
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.96-97
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    • 2012
  • In nitride and oxide film deposition, sputtered metals react with nitrogen or oxygen gas in a vacuum chamber to form metal nitride or oxide films on a substrate. The physical properties of sputtered films (metals, oxides, and nitrides) are strongly influenced by magnetron plasma density during the deposition process. Typical target power densities on the magnetron during the deposition process are ~ (5-30) W/cm2, which gives a relatively low plasma density. The main challenge in reactive sputtering is the ability to generate a stable, arc free discharge at high plasma densities. Arcs occur due to formation of an insulating layer on the target surface caused by the re-deposition effect. One current method of generating an arc free discharge is to use the commercially available Pinnacle Plus+ Pulsed DC plasma generator manufactured by Advanced Energy Inc. This plasma generator uses a positive voltage pulse between negative pulses to attract electrons and discharge the target surface, thus preventing arc formation. However, this method can only generate low density plasma and therefore cannot allow full control of film properties. Also, after long runs ~ (1-3) hours, depends on duty cycle the stability of the reactive process is reduced due to increased probability of arc formation. Between 1995 and 1999, a new way of magnetron sputtering called HIPIMS (highly ionized pulse impulse magnetron sputtering) was developed. The main idea of this approach is to apply short ${\sim}(50-100){\mu}s$ high power pulses with a target power densities during the pulse between ~ (1-3) kW/cm2. These high power pulses generate high-density magnetron plasma that can significantly improve and control film properties. From the beginning, HIPIMS method has been applied to reactive sputtering processes for deposition of conductive and nonconductive films. However, commercially available HIPIMS plasma generators have not been able to create a stable, arc-free discharge in most reactive magnetron sputtering processes. HIPIMS plasma generators have been successfully used in reactive sputtering of nitrides for hard coating applications and for Al2O3 films. But until now there has been no HIPIMS data presented on reactive sputtering in cluster tools for semiconductors and MEMs applications. In this presentation, a new method of generating an arc free discharge for reactive HIPIMS using the new Cyprium plasma generator from Zpulser LLC will be introduced. Data (or evidence) will be presented showing that arc formation in reactive HIPIMS can be controlled without applying a positive voltage pulse between high power pulses. Arc-free reactive HIPIMS processes for sputtering AlN, TiO2, TiN and Si3N4 on the Applied Materials ENDURA 200 mm cluster tool will be presented. A direct comparison of the properties of films sputtered with the Advanced Energy Pinnacle Plus + plasma generator and the Zpulser Cyprium plasma generator will be presented.

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Geochemical characteristics of a LILW repository I. Groundwater (중.저준위 방사성 폐기물 처분부지의 지구화학 특성 I. 지하수)

  • Choi, Byoung-Young;Kim, Geon-Young;Koh, Yong-Kwon;Shin, Seon-Ho;Yoo, Si-Won;Kim, Doo-Haeng
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.6 no.4
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    • pp.297-306
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    • 2008
  • This study was carried out to identify the characteristics of hydrochemistry controlling groundwater chemical condition in a repository site of Gyeongju. For this study, 12 bore holes of all monitoring bore holes in the study area were selected and total 46 groundwater samples were collected with depth. In addition, 3 surfacewater samples and 1 seawater sample were collected. For water samples, cations and anions were analyzed. The environmental isotopes(${\delta}^{18}O-{\delta}D$, Tritium, ${\delta}^{13}C,\;{\cdot}{\delta}^{34}S$) were also analyzed to trace the origin of water and solutes. The result of ${\delta}^{18}O\;and\;{\delta}D$ analysis showed that surface water and groundwater were originated from precipitation. Tritium concentrations of groundwater decreased with depth but high concentrations of tritium indicated that groundwater was recharged recently. The results of ion and correlation analysis showed that groundwater types of the study area were represented by Ca-Na-$HCO_3$ and Na-Cl-$SO_4$, which was caused by sea spray and water-rock interaction. Especially, high ratio of Na content in groundwater resulted from ion exchange. For redox condition of groundwater, the values of DO and Eh decreased with depth, which indicated that reducing condition was formed in deeper groundwater. In addtion, high concentration of Fe and Mn showed that redox condition of groundwater was controlled by the reduction of Fe and Mn oxides.

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Hydrogeological characteristics of the LILW disposal site (처분부지의 수리지질 특성)

  • Kim, Kyung-Su;Kim, Chun-Soo;Bae, Dae-Seok;Ji, Sung-Hoon;Yoon, Si-Tae
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.6 no.4
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    • pp.245-255
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    • 2008
  • Korea Hydro and Nuclear Power Company(KHNP) conducted site investigations for a low and intermediate-level nuclear waste repository in the Gyeong Ju site. The site characterization work constitutes a description of the site, its regional setting and the current state of the geosphere and biosphere. The main objectives of hydogeological investigation aimed to understand the hydrogeological setting and conditions of the site, and to provide the input parameters for safety evaluation. The hydogeological characterization of the site was performed from the results of surface based investigations, i.e geological mapping and analysis, drilling works and hydraulic testing, and geophysical survey and interpretation. The hydro-structural model based on the hydrogeological characterization consists of one-Hydraulic Soil Domain, three-Hydraulic Rock Domains and five-Hydraulic Conductor Domains. The hydrogeological framework and the hydraulic values provided for each hydraulic unit over a relevant scale were used as the baseline for the conceptualization and interpretation of flow modeling. The current hydrogeological characteristics based on the surface based investigation include some uncertainties resulted from the basic assumption of investigation methods and field data. Therefore, the reassessment of hydrostructure model and hydraulic properties based on the field data obtained during the construction is necessitated for a final hydrogeological characterization.

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A Study on Designing Method of VoIP QoS Management Framework Model under NGN Infrastructure Environment (NGN 기반환경 에서의 VoIP QoS 관리체계 모델 설계)

  • Noh, Si-Choon;Bang, Kee-Chun
    • Journal of Digital Contents Society
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    • v.12 no.1
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    • pp.85-94
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    • 2011
  • QoS(Quality of Service) is defined as "The collective effect of service performance which determines the degree of satisfaction of a user of the service" by ITU-T Rec. E.800. While the use of VoIP(Voice Over Internet Protocol) has been widely implemented, persistent problems with QoS are a very important sue which needs to be solved. This research is finding the assignment of VoIP QoS to deduct how to manage the control system and presenting the QoS control process and framework under NGN(Next Generation Network) environment. The trial framework is the modeling of the QoS measurement metrics, instrument, equipment, method of measurement, the series of cycle & the methodology about analysis of the result of measurement. This research underlines that the vulnerability of the VoIP protocol in relation to its QoS can be guaranteed when the product quality and management are controlled and measured systematically. Especially it's very important time to maintain the research about VoIP QoS measurement and control because the big conversion of new network technology paradigm is now spreading. In addition, when the proposed method is applied, it can reduce an overall delay and can contribute to improved service quality, in relation to signal, voice processing, filtering more effectively.

A Study on CFD Analysis to Investigate the Effects of Different Feed Rate into the High Temperature H2SO4 Transferring Pump at Fixed Frequency

  • Choi, Jung-Sik;Choi, Jae-Hyuk
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.20 no.3
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    • pp.304-311
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    • 2014
  • In this study, to apply hydrogen energy to ship engine and to generate effective hydrogen production, we investigated the effects of high temperature $H_2SO_4$ feed rate and cooling water rate to pump parts with fixed frequency needed to reciprocate motion and a simulation was conducted at each condition. In the fixed frequency and cooling water inlet flow rate of 0.5 Hz and 3.9 kg/s, we changed the high temperature $H_2SO_4$ flow rate to 47.46 kg/s (it is 105 % of 45.2 kg/s), 49.72 kg/s (110 %), and 51.98 kg/s (115 %). Also, at 0.5 Hz and 45.2 kg/s of frequency and high temperature $H_2SO_4$ flow, the thermal hydraulic analysis was performed at the condition of 95 % (3.705 kg/s), 90 % (3.51 kg/s), and 85 % (3.315 kg/s). In overall simulation cases, the physical properties of materials are more influential to the temperature increase in the pump part rather than the changes on the feed rate of high temperature $H_2SO_4$ and cooling water. A continuous operation of pump was also capable even if the excess feed of high temperature $H_2SO_4$ of about 15 % or the less feed of cooling water of about 15 % were performed, respectively. When the increasing feed of high temperature $H_2SO_4$ of up to 5 %, 10 %, and 15 % were compared with base flow (45.2 kg/s), the deviation of time period rose to a certain temperature and ranged from 0 to 4.5 s in the same position (same material). In case of cooling water, the deviation of time period rose to a certain temperature and ranged from 0 to 5.9 s according to the decreasing feed changes of cooling water at 5 %, 10 %, and 15 % compared to a base flow (3.9 kg/s). Finally, the additional researches related to the two different materials (Teflon and STS for Pitch and End-plate), which are concerned about the effects of temperature changes to the parts contacting different materials, are needed, and we have a plan to conduct a follow-up study.

Computational Analysis for a Molten-salt Electrowinner with Liquid Cadmium Cathode (액체 카드뮴 음극을 사용한 용융염 전해제련로 전산해석)

  • Kim, Kwang-Rag;Jung, Young-Joo;Paek, Seung-Woo;Kim, Ji-Yong;Kwon, Sang-Woon;Yoon, Dal-Seong;Kim, Si-Hyung;Shim, Jun-Bo;Kim, Jung-Gug;Ahn, Do-Hee;Lee, Han-Soo
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.8 no.1
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    • pp.1-7
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    • 2010
  • In the present work, an electrowinning process in the LiCl-KCl/Cd system is considered to model and analyze the electrotransport of the actinide and rare-earth elements. A simple dynamic modeling of this process was performed by taking into account the material balances and diffusion-controlled electrochemical reactions in a diffusion boundary layer at an electrode interface between the molten salt electrolyte and liquid cadmium cathode. The proposed modeling approach was based on the half-cell reduction reactions of metal chloride occurring on the cathode. This model demonstrated a capability for the prediction of the concentration behaviors, a faradic current of each element and an electrochemical potential as function of the time up to the corresponding electrotransport satisfying a given applied current based on a galvanostatic electrolysis. The results of selected case studies including five elements (U, Pu, Am, La, Nd) system are shown, and a preliminary simulation is carried out to show how the model can be used to understand the electrochemical characteristics and provide better information for developing an advanced electrowinner.