• Title/Summary/Keyword: Marker and Cell method

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Quantification of the Bioactive Components of the Rhizomes of Curcuma wenyujin and Assessment of Its Anti-inflammatory Effect in Benign Prostatic Hyperplasia-1 Cells

  • Seo, Chang-Seob;Park, Eunsook;Lee, Mee-Young;Shin, Hyeun-Kyoo
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.238-243
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    • 2019
  • In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini $C_{18}$ columns ($250mm{\times}4.6mm$, $5{\mu}m$) at $40^{\circ}C$ by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin $E_2$ ($PGE_2$) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were $0.10-0.32{\mu}g/mL$ and $0.30-0.98{\mu}g/mL$, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 - 5.92 mg/g, 4.72 - 4.86 mg/g, and 1.06 - 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed $PGE_2$ levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit antiinflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.

Construction of an expression vector with SV40 DNA in a mammalian cell (SV 40 DNA를 이용한 포유동물의 유전자 운반체 개발)

  • 정민혜;김상해;전희숙;노현모
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.165-172
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    • 1987
  • An expression vector in a mammalian cell was constructed using the origin of replication (OR) and the promoters of SV40. The plasmid pSVOE was constructed by inserting SV40 DNA fragment (1, 118bp) containing SV40 OR and promoters into pBR322-1, and then a multiple cloning sequence was inserted at the immediate downstream of the late promoter of SV40 in the pSVOE vector. The plasmid was named pSVML. As a selection marker, thymidine kinase gene of herpes simplex virus with its promoter was inserted into EcoRI site of pSVML and the recombinant was named pSVML-TKp. To test the expression capacity of foreigen gene inserted at the multiple cloning site of pSVML, the thymidine kinase gene without its own promoter was inserted at the BamHI site of pSVML. The recombinant was named pSVML-TK. These plasmids, pSVML-TKp and pSVML-TK, were transfected into COS cells with calcium phosphate precipitation method. The thymidine kinase activity was significantly increased in both transfected cells.

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A Numerical Calculation of Viscous Flow around a Hydrofoil Advancing beneath the Free Surface (자유수면 아래서 전진하는 수중익 주위의 점성유동 해석)

  • J.J. Park;S.M. Jeong;Y.G. Lee;S.H. Lee;S.W. Hong
    • Journal of the Society of Naval Architects of Korea
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    • v.32 no.3
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    • pp.72-82
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    • 1995
  • In the present paper, flow characteristics and free surface waves generated by a submerged hydrofoil advancing with an uniform speed are calculated. Using a numerical method based on a MAC(Marker And Cell) method, the Navier-Stokes and the continuity equations are solved to simulate flow fields around the hydrofoil. Computations are carried out in a rectangular grid system in which grids are concentrated near the foil and the free surface to improve numerical accuracies. Viscous flow phenomenas including pressure distributions are computed. Moreover, the influences of submerged depths upon the generated wave profiles and the wave breaking phenomena are also investigated. Experiments are performed at the towing tank of Inha University to measure free surface wave elevations due to the advancing hydrofoil. The computational results are compared with the present and the other available experimental data to show the accuracy of the numerical method developed.

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Improvement of Histopathological Sample Preparation by Employing Microwave Heating Method on Frozen Section Specimens

  • Ahn, Seung-Ju
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.361-368
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    • 2007
  • Biological samples can be fixed either by chemical method by using chemical solution or physical methods by using heat treatment. The problem in traditional heat fixation is unsatisfactory quality due to uneven heat conduction in specimen and loss of inner cell contents. Chemical fixation method also bears several intrinsic problems like the limit in specimen size, time consumption in fixative impregnation, and loss of low molecular weight cell components. These factors deteriorate the quality of fixed specimen, thus limit the magnification and contrast of tissue pictures. Microwave heat has been reported to be a good alternative to current chemical methods to overcome these problem. In this study, we tried to introduce the microwave energy method to routine fixation work in hospital. We replaced chemical fixative with saline to provide moderate reaction condition, and used frozen section to reduce time for sample preparation. Temperature was measured at each experiment. The fixation of rat kidney tissue with 2.45 GHz electromagnetic wave and saline showed similar result to the control group fixed with traditional chemical method. Human tumor tissue fixed with 2.45 GHz electromagnetic in frozen section was improved in terms of histochemistry of PAS and immunohistochemistry of tumor marker like cytokeratin. Total turnaround time was reduced from $24\sim38$ h to to $2\sim4$ h. In conclusion, the quality of samples prepared by microwave heating method was at least as good as that of traditional method. If the condition for the fixation of different specimens is standardized, this new method could be applied to routine work in hospital, and could save working time as well.

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Effect of complex mechanical stimuli for MC3T3-E1 cells (MC3T3-E1 세포에 대한 복합 기계적 자극의 영향)

  • Kang, Kyung-Shin;Lee, Seung-Jae;Cho, Dong-Woo
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1700-1703
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    • 2008
  • The purpose of this study was to examine the effects of various mechanical stimuli for MC3T3-E1 cells. Among the several mechanical stimulations, we focused on compressive stain and ultrasound. In this study, we developed a bioreactor capable of applying controlled stimuli to scaffolds. PLLA/PCL scaffold was fabricated by using salt-leaching method. We performed dynamic cell culture using preosteoblasts MC3T3-E1 cells with 1MHz, 30mW/cm2 ultrasound and 10% of compressive strain. Result of CCK-8 analysis at 1, 4, 7, 10 days showed that mechanical stimuli had no significant effect for cell proliferation. However, those stimuli influenced ALP(Alkaline phopatase) activity, which is one of differentiation marker.

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A Study on the Bow Wave Characteristics for the Resistance-Minimized Hull Form of Small Fishing Boat (저항최소화 소형 어선선형의 선수파 특성에 관한 연구)

  • Yu, Jin-Won;Lee, Young-Gill
    • Journal of the Society of Naval Architects of Korea
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    • v.45 no.2
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    • pp.124-131
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    • 2008
  • This paper presents the analysis of resistance performance and bow waves for the resistance-minimized hull form of small fishing boat by using numerical simulations and model tests. The resistance-minimized hull form is developed from an original hull form which is selected from existing small fisher boats in our country. In order to estimate the resistance performance for the original and the developed hull form, several numerical simulations and model tests are carried out. Marker and Cell(MAC) method and Marker-Density method are adopted to simulate the free-surface bow waves around advancing hull surface. The results of numerical simulations are compared with the model tests in towing tank. The results show that the resistance performance of the resistance-minimized hull form is improved than that of the original hull form. The results of this study will be a good guide to the hull form development of small fishing boats in future.

Effect of p38 inhibitor on the proliferation of chicken muscle stem cells and differentiation into muscle and fat

  • Minkyung, Ryu;Minsu, Kim;Hyun Young, Jung;Cho Hyun, Kim;Cheorun, Jo
    • Animal Bioscience
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    • v.36 no.2
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    • pp.295-306
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    • 2023
  • Objective: Inhibiting the p38 mitogen-activated protein kinase (MAPK) signaling pathway delays differentiation and increases proliferation of muscle stem cells in most species. Here, we aimed to investigate the effect of p38 inhibitor (p38i) treatment on the proliferation and differentiation of chicken muscle stem cells. Methods: Chicken muscle stem cells were collected from the muscle tissues of Hy-line Brown chicken embryos at embryonic day 18, then isolated by the preplating method. Cells were cultured for 4 days in growth medium supplemented with dimethyl sulfoxide or 1, 10, 20 μM of p38i, then subcultured for up to 4 passages. Differentiation was induced for 3 days with differentiation medium. Each treatment was replicated 3 times. Results: The proliferation and mRNA expression of paired box 7 gene and myogenic factor 5 gene, as well as the mRNA expression of myogenic differentiation marker gene myogenin were significantly higher in p38i-treated cultures than in control (p<0.05), but immunofluorescence staining and mRNA expression of myosin heavy chain (MHC) were not significantly different between the two groups. Oil red O staining of accumulated lipid droplets in differentiated cell cultures revealed a higher lipid density in p38i-treated cultures than in control; however, the expression of the adipogenic marker gene peroxisome proliferator activated receptor gamma was not significantly different between the two groups. Conclusion: p38 inhibition in chicken muscle stem cells improves cell proliferation, but the effects on myogenic differentiation and lipid accumulation require additional analysis. Further studies are needed on the chicken p38-MAPK pathway to understand the muscle and fat development mechanism.

Clinical Utility of Haptoglobin in Combination with CEA, NSE and CYFRA21-1 for Diagnosis of Lung Cancer

  • Wang, Bing;He, Yu-Jie;Tian, Ying-Xing;Yang, Rui-Ning;Zhu, Yue-Rong;Qiu, Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9611-9614
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    • 2014
  • Purpose: To investigate the clinical value in lung cancer of a combination of four serum tumor markers, haptoglobin (Hp), carcinoembryonic antigen (CEA), neuron specific enolase (NSE) as well as the cytokeratin 19 fragment (CYFRA21-1). Materials and Methods: Serum Hp (with immune-turbidimetric method), CEA, NSE, CYFRA21-1 (with chemiluminescence method) level were assessed in 193 patients with lung cancer, 87 patients with benign lung disease and 150 healthy controls. Differences of expression were compared among groups, and joint effects of these tumor markers for the diagnosis of lung cancer were analyzed. Results: Serum tumor marker levels in patients with lung cancer were obviously higher than those with benign lung disease and normal controls (p<0.01). The sensitivities of Hp, CEA, NSE and CYFRA21-1 were 43.5%, 40.9%, 23.3% and 41.5%, with specificities of 90.7%, 99.2%, 97.9% and 97.9%. Four tumor markers combined together could produce a positive detection rate of 85.0%, significantly higher than that of any single test. With squamous carcinomas, the positive detection rates with Hp and CYFRA21-1 were higher than that of other markers. In the adenocarcinoma case, the positive detection rate of CEA was higher than that of other markers. For small cell carcinomas, the positive detection rate of NSE was highest. The area under receiver operating characteristic curve ($AUC^{ROC}$) of Hp in squamous carcinoma (0.805) was higher than in adenocarcinoma (0.664) and small cell carcinoma (0.665). Conclusions: Hp can be used as a new serum tumor marker for lung cancer. Combination detection of Hp, CEA, NSE and CYFRA21-1 could significantly improve the sensitivity and specificity in diagnosis of lung cancer, and could be useful for pathological typing.

In Situ Single Cell Monitoring by Isocyanide-Functionalized Ag and Au Nanoprobe-Based Raman Spectroscopy

  • Lee, So-Yeong;Jang, Soo-Hwa;Cho, Myung-Haing;Kim, Young-Min;Cho, Keun-Chang;Ryu, Pan Dong;Gong, Myoung-Seon;Joo, Sang-Woo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.904-910
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    • 2009
  • The development of effective cellular imaging requires a specific labeling method for targeting, tracking, and monitoring cellular/molecular events in the living organism. For this purpose, we studied the cellular uptake of isocyanide-functionalized silver and gold nanoparticles by surface-enhanced Raman scattering (SERS). Inside a single mammalian cell, we could monitor the intracellular behavior of such nanoparticles by measuring the SERS spectra. The NC stretching band appeared clearly at ${\sim}2,100cm^{-1}$ in the well-isolated spectral region from many organic constituents between 300 and 1,700 or 2,800 and $3,600cm^{-1}$. The SERS marker band at ${\sim}2,100cm^{-1}$ could be used to judge the location of the isocyanide-functionalized nanoparticles inside the cell without much spectral interference from other cellular constituents. Our results demonstrate that isocyanide-modified silver or gold nanoparticle-based SERS may have high potential for monitoring and imaging the biological processes at the single cell level.

Study of Natural Convection of Magnetic Fluid in Cubic Cavity (정방형관 내에서 자성유체의 자연대류현상에 대한 수치적 연구)

  • Seo, Jae-Hyeong;Lee, Moo-Yeon;Seo, Lee-Soo
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.37 no.7
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    • pp.637-646
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    • 2013
  • This study aims to numerically investigate the natural convection characteristics of a magnetic fluid in a cubic cavity. The governing equations of the magnetic fluid are solved using the Generalized-Simplified Marker and Cell Method (GSMAC). The natural convection and heat transfer characteristics of the magnetic fluid were analyzed by varying the intensity and direction of the magnetic field. As a result, it was found that the natural convection characteristics were controlled by the intensity and direction of the magnetic field, and the mean Nusselt numbers were minimized at a vertical intensity of H=-4000 and horizontal intensity of H=12000 of the magnetic field. In addition, the mean Nusselt numbers increased with the intensities of the magnetic field, regardless of the direction of the magnetic field.