• Title/Summary/Keyword: 형광분석법

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The study on cytotoxicity of cytokines produced by the activated human NKT cells on neuroblastoma (활성화된 자연살상 T 세포(NKT)에서 생성된 사이토카인에 의한 신경모세포종의 세포독성에 관한 연구)

  • Cho, Jin Young;Yoon, Young Wook;Yoon, Hyang Suk;Kim, Jong Duk;Choi, Du Young
    • Clinical and Experimental Pediatrics
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    • v.49 no.4
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    • pp.439-445
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    • 2006
  • Purpose : ${\alpha}$-Galactosylceramide (${\alpha}$-GalCer)-stimulated human $V{\alpha}24$ natural killer T (NKT) cells exert antitumor activity against some leukemia in a CD1d dependent and TCR-mediated manner, but could not kill CD1d - negative neuroblastoma (NB) cells. There are few reports about the direct antitumor effect of highly secreted cytokines by these cells on activation. In this study, using a cell-free supernatant (SPN) collected from plate bound hCD1d/${\alpha}$ GalCer tetramers-stimulated NKT cells, we examined whether they could be helpful in the immunotherapeutic treatment of NB. Methods : Cells were cultured in IMDM. The cytokines produced by NKT cells were measured with Cytometric Bead Array (CBA) analysis. Cell viability was evaluated by calcein-AM fluorescence with digital image microscopy scanning (DIMSCAN). The percentage of specific apoptosis was calculated by flow cytometric detection of apoptosis using annexin V and 7-AAD. Results : The activated NKT cells secreted high levels of IL-2, INF-${\gamma}$, TNF-${\alpha}$. The SPN was significantly cytotoxic against four out of eight tested NB cell lines, through mainly apoptosis as evidenced by annexin-V staining and inhibition with the pretreatment of pancaspase blocker. This apoptosis was significantly inhibited when anti-TNF-${\alpha}$ and anti-IFN-${\gamma}$ neutralizing mAbs were used separately and it was completely abolished when the two mAbs were combined. Conclusion : IFN-${\gamma}$ and TNF-${\alpha}$ produced by NKT cells could exert synergistically direct antitumor activity through apoptosis on some NB cell lines.

Usefulness of serum procalcitonin test for the diagnosis of upper urinary tract infection in children (소아 상부 요로감염의 진단을 위한 혈청 procalcitonin 검사의 유용성)

  • Kim, Dong Wook;Chung, Ju Young;Koo, Ja Wook;Kim, Sang Woo;Han, Tae Hee
    • Clinical and Experimental Pediatrics
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    • v.49 no.1
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    • pp.87-92
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    • 2006
  • Purpose : It is difficult to make a distinction between lower urinary tract infection(UTI) and acute pyelonephritis(APN) during the acute phase of febrile UTI due to nonspecific clinical symptoms and laboratory findings, especially among young children. We measured the serum procalcitonin(PCT) in children with UTI to distinguish between acute pyelonephritis and lower UTI, and to determine the accuracy of PCT measurement compared with other inflammatory markers. Methods : Serum samples were taken from children who admitted with unexplained fever or were suspected of having UTI. 51 children(mean $12.2{\pm}11.4$ months) were enrolled in this study. Leukocyte counts, erythrocyte sedimentation rates(ESR) and C-reactive protein(CRP) were also measured. Renal parenchymal involvement was assessed by $^{99m}Tc$ DMSA scintigraphy in the first 7 days after admission. PCT was measured by immunoluminometric assay. Results : PCT values were significantly correlated with the presence of renal defects in children with UTI(n=16)($5.06{\pm}12.97{\mu}g/L$, P<0.05). However, PCT values were not significantly different between children with UTI without renal damage(n=18) and children without UTI(n=17). Using a cutoff of $0.5{\mu}g/L$ for PCT and 20 mm/hr for ESR, 20 mg/L for CRP, sensitivity and specificity in distinguishing between UTI with and without renal involvement were 81.3 percent and 88.9 percent for PCT 87.5 percent and 72.2 percent for ESR, and 87.5 percent and 55.6 percent for CRP, respectively. Positive and negative predictive values were 86.7 percent and 84.2 percent for PCT and 60.9 percent and 81.8 percent for CRP, respectively. Conclusion : In febrile UTI, PCT values were more specific than CRP, ESR and leukocyte count for the identification of patients who might develop renal defects.

The Role of Heme Oxygenase-1 in Lung Cancer Cells (폐암세포주에서 Heme Oxygenase-1의 역할)

  • Jung, Jong-Hoon;Kim, Hak-Ryul;Kim, Eun-Jung;Hwang, Ki-Eun;Kim, So-Young;Park, Jung-Hyun;Kim, Hwi-Jung;Yang, Sei-Hoon;Jeong, Eun-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.304-313
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    • 2006
  • Background : Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes the oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. The current evidence has indicated a critical role of HO-1 in cytoprotection and also in other, more diverse biological functions. It is known that the high expression of HO-1 occurs in various tumors, and that HO-1 has an important role in rapid tumor growth because of its antioxidative and antiapoptotic effects. Therefore, the role of HO-1 was analyzed in human lung cancer cell lines, and especially in the A549 cell line. Material and Methods : Human lung cancer cell lines, i.e., A549, NCI-H23, NCI-H157 and NCI-H460, were used for this study. The expression of HO-1 in the untreated state was defined by Western blotting. ZnPP, which is the specific HO inhibitor we used, and the viability of cells were tested for by conducting MTT assaysy. The HO enzymatic activity, as determined via the bilirubin level, was also indirectly measured. Moreover, the generation of intracellular hydrogen peroxide (H2O2) was monitored fluorimetrically with using a scopoletin-horse radish peroxidase (HRP) assay and 2',7'-dichlorofluorescein diacetate (DCFH-DA). We have also transfected small HO-1 interfering RNA (siRNA) into A549 cells, and the apoptotic effects were evaluated by flow cytometric analysis and Western blotting. Results : The A549 cells had a greater expression of HO-1 than the other cell lines, whereas ZnPP significantly decreased the viability of the A549 cells more than the viability of the other lung cancer cells in a dose-dependant fashion. Consistent with the viability, the HO enzymatic activity also was decreased. Moreover, intracellular H2O2 generation via ZnPP was induced in a dose-dependent manner. Apoptotic events were, then induced in the HO-1 siRNA transfected A549 cells. Conclusion : HO-1 provides new important insights into the possible molecular mechanism of the antitumor therapy in lung cancer.

The optimal conditions to improve retrovirus-mediated transduction efficiency to NIH 3T3 cells (레트로바이러스(retrovirus)의 NIH 3T3 세포로의 유전자 전달효율을 증가시키기 위한 적절한 조건들)

  • Lee, Jun Ah;Lee, Kang-Min;Lee, Hyun Jae;Lee, Yun Jeong;Kim, Dong Ho;Lim, Jung Sub;Park, Kyung-Duk
    • Clinical and Experimental Pediatrics
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    • v.50 no.10
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    • pp.1011-1017
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    • 2007
  • Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

Analysis of the Spatial Dose Rates during Dental Panoramic Radiography (치과 파노라마 촬영에서 공간선량률 분석)

  • Ko, Jong-Kyung;Park, Myeong-Hwan;Kim, Yongmin
    • Journal of radiological science and technology
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    • v.39 no.4
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    • pp.509-516
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    • 2016
  • A dental panoramic radiography which usually uses low level X-rays is subject to the Nuclear Safety Act when it is installed for the purpose of education. This paper measures radiation dose and spatial dose rate by usage and thereby aims to verify the effectiveness of radiation safety equipment and provide basic information for radiation safety of radiation workers and students. After glass dosimeter (GD-352M) is attached to direct exposure area, the teeth, and indirect exposure area, the eye lens and the thyroid, on the dental radiography head phantom, these exposure areas are measured. Then, after dividing the horizontal into a $45^{\circ}$, it is separated into seven directions which all includes 30, 60, 90, 120 cm distance. The paper shows that the spatial dose rate is the highest at 30 cm and declines as the distance increases. At 30 cm, the spatial dose rate around the starting area of rotation is $3,840{\mu}Sv/h$, which is four times higher than the lowest level $778{\mu}Sv/h$. Furthermore, the spatial dose rate was $408{\mu}Sv/h$ on average at the distance of 60 cm where radiation workers can be located. From a conservative point of view, It is possible to avoid needless exposure to radiation for the purpose of education. However, in case that an unintended exposure to radiation happens within a radiation controlled area, it is still necessary to educate radiation safety. But according to the current Medical Service Act, in medical institutions, even if they are not installed, the equipment such as interlock are obliged by the Nuclear Safety Law, considering that the spatial dose rate of the educational dental panoramic radiography room is low. It seems to be excessive regulation.

Seroepizootiological Study on Bovine Leucosis in Korea (한국(韓國)에서의 소백혈병(白血病)의 혈청역학적(血淸疫學的) 연구(硏究))

  • Jun, Moo-Hyung;Chung, Un-Ik;Lee, Chang-Ku;Baig, Soon-Yong;Lim, Chang-Hyeong
    • Korean Journal of Veterinary Research
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    • v.22 no.2
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    • pp.175-185
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    • 1982
  • Since bovine lymphosarcoma causes considerable economic loss to the dairy industry, seroepidemiological survey on bovine leucosis virus (BLV) was carried out for the dairy herds throughout the country to observe the epidemiological situation of the disease by using immunodiffusion test. Attempts were simultaneously made to detect bovine leucosis virus in the lymphocytes from BLV antibody-positive cattle by means of fluorescent antibody techniques, syncytium assay and electron microscopy. In immunodiffusion test for BLV antibody in 2003 heads of dairy cattle selected randomly from 164 herds, the prevalence of positive reactors by regions were 37.8% in Central, 27.2% in Honam (Southwest), 28.0% in Youngnam (Southeast) and 25.2% in Youngdong (East coast)and averaging 29.7%. By provinces, Chungcheong appeared the highest prevalence of BLV antibody carriers (41.8%), while Jeonbug revealed the lowest incidence rate (24.4%). When the results of serological studies were analyzed by age groups and the sizes of herds, the number of reactors increased gradually with the advance in the age of cattle and the herd size. The highest rate of BLV carriers was found in the ages between 6 and 8 years, and in the size of herds with 20 to 50 heads. One hundred and seventeen breeding bulls from the central regions were tested for BLV antibody. Four out of 70 bulls (5.7%) of Korean cattle and 14 out of 39 bulls (35.9%) of Holstein were reactive for BLV antigens. Of 164 dairy herds examined, 17 herds (10.4%) have no BLV antibody-positive cattle, while 42 herds (25.6%) were included in the range of 20 to 40% of the positive rate and 10 herds (6.1%) in the range of over 80% of the rate. When the lymphocytes from the BLV antibody carrying cattle were cultured in the presence of phytohemagglutinin and stained with FITC-conjugated sheep anti-BLV serum, 8 out of 11 cases (72.7%) of BLV positive cattle revealed specific fluorescence for BLV in the lymphocytes. In syncytium assay of the peripheral lymphocytes of the cattle, 5 out of 7 (71.4%) lymphocytes from BLV antibody carriers induced syncytia in the indicators of bovine embryonic splenic cells. The cultured lymphocytes were examined with an electron microscope to detect the BLV particles. Two out of 6 specimens (33.3%) from the reactors showed the typical type C virus with the size of 90 to 110 nm around microvilli and in intracytoplasmic vacuoles.

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Analysis of Signal Properties in accordance with electrode area of x-ray conversion material (X선 검출 물질의 전극 면적에 따른 신호특성 분석)

  • Jeon, S.P.;Kim, S.H.;CHO, K.S.;Jung, S.H.;Park, J.K.;Kang, S.S.;Han, Y.H.;Kim, K.S.;Mun, C.W.;Nam, S.H.
    • Journal of the Korean Society of Radiology
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    • v.4 no.1
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    • pp.5-9
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    • 2010
  • In recent, a digital x-ray detector attracted worldwide attention and there are many studies to commercialize. There are two methods in digital x-ray detector. This method is an Indirect method and Direct method. This study is to see the differences between the digital x-ray detector based on a-Se used in the existing indirect conversion method and an x-ray conversion material that has better SNR(Signal-to-noise ratio) and property than the a-Se. To solve the problem that is difficult to make a large area film using Screen-Print method, we used a Screen-Print method. In this study, we used a polyclystal $HgI_2$ as x-ray conversion material and a sample thickness is $150{\mu}m$ and an area is $3cm{\times}3cm$. ITO(Indium-Tin-Oxide) electrode was used as top electrode using a Magnetron Sputtering System and each area is $3cm{\times}3cm$, $2cm{\times}2cm$ and $1cm{\times}1cm$ and then we evaluated darkcurrent, sensitivity and SNR of the $HgI_2$ film are measured, then we evaluated the electrical properties. And we used a current integration mode when I-V test. This experiment shows that the sensitivity increases in accordance with the area of the electrode but the SNR is decreased because of the high darkcurrent. Through fabricating of various thicknesses and optimal electrodes, we will optimize SNR in the future work.