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Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • 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.

Effect of Unilateral Renal Perfusion of Cyclosporine and Mitomycin on Rat's Kidney (Cyclosporine과 Mitomycin의 일측성 신관류로 초래되는 백서 신병변에 관한 연구)

  • Baek Seung In;Lim Hyun Suk;Shin Weon Hye;Ko Cheol Woo;Koo Ja Hoon;Kwak Jung Sik
    • Childhood Kidney Diseases
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    • v.2 no.2
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    • pp.138-144
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    • 1998
  • Purpose : The use of cyclosporine and mitomycin in various immunologic or neoplastic disorders has been known to cause wide-ranged nephrotoxic effects including thrombotic microangiopathy. However, the mechanism of nephrotoxicity of these drugs has not been studied adequately, so that present experimental study has been undertaken to find out whether these drugs can cause direct damage to the kidney and to clarify the pathogenetic mechanism of nephrotoxic effect of these drugs. Materials and methods : Sprague-Dawley rats weighing 250-300 gm were used for experimental animals and unilateral renal perfusion technique, modified from the method described by Hoyer et al was used. Isolation of left kidney from systemic circulation was made by clamping aorta and left renal vein and a hole was punctured in the anterior wall of the left renal vein. Cyclosporine (2.5 mg in 4 ml solution) and mitomycin (1.6 mg in 4ml solution) were infused through left renal artery and normal saline was used in control rats. Forty-eight hours after infusion of the drugs, animals were sacrificed and left kidney removed and processed for histologic examination. Total ischemic time of left kidney was less than 15 minutes: Results : Cyclosporine-perfused group showed severe swelling of glomerular endothelial ceil along with swelling of glomerular epithelial cell and interstitial vascular endothelial cell. Mitomycin-perfused group also showed severe swelling of glomerular endothelial and epithelial cells. And in addition to these findings, they demonstrated platelets aggregation, swelling and degranulation of platelets and fibrin accumulation in some of the capillaries, indicating occurrance of thrombotic microangiopathy. Conclusion : present experiment indicates that cyclosporine and mitomycin can cause direct toxic injury to renal endothelial cell. And this direct toxic damage to endothelial cell seems to be an important initiating event for the development of thrombotic microangiopathy.

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Usefulness of Flow Composite Image in Raynaud Scan ($^{201}Tl$) ($^{201}Tl$을 이용한 레이노 검사에서 동적 Composite 영상의 유용성)

  • Kim, Dae-Yeon;Shin, Gyoo-Seol;Oh, Eun-Jung;Kim, Gun-Jae
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.1
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    • pp.101-104
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    • 2010
  • Purpose: Raynaud scan is divided to flow, blood pool and local-delay image. Usually, we evaluate comparison through blood pool and local-delay image. We will evaluate about usability when comparative observe blood image and local-delay image in Raynaud scan that used $^{201}Tl$ as making flow image to one sheet of images. Materials and Methods: We have selected 29 Raynaud phenomenon patients aged 14~68 years who visited department of vascular surgery between Feb. 2008 and Aug. 2009. An intravenous injection $^{201}Tl$ of 111 MBq (3 mCi) to opposite side diagonal line limbs above an internal auditing department. Equipment used Philips gamma camera forte A-Z, and collimator used LEHR. Matrix size set up to each $64{\times}64$, $128{\times}128$, $256{\times}256$ and zoom factor used to full field. Protocol of dynamic is 2 second to 155 frames. Blood pool and delay count to 300 second. We set up ROI by a foundation to data acquired in PEGASYS processing program. Each results were analyzed with the SPSS 12.0 statistical software. Results: Each averages of count ratio (Rt / Lt) to have been given at composite image, a blood pool image, delay images analyzed at Raynaud phenomenon patients is $1.25{\pm}0.39$, $1.20{\pm}0.33$, $1.11{\pm}0.17$. The sample analysis results of blood pool image and delay image contented itself with p<0.029. Also, there don't have been each difference, and blood pool image, delay image regarding composite image was able to know. Conclusion: We were able to give help for comparison to evaluate a blood pool image and a local delay image at the Raynaud scan which used $^{201}Tl$ while making a flow image to one sheet image. Identification to be visual too was possible. If you are proceeded a researcher that there was further depth, you are more appropriate for, and you may get useful information.

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Relations between Polymorphism of NRAMP1 Gene and Susceptibility to Pulmonary Tuberculosis (NRAMP1 유전자 다형성과 폐결핵의 감수성과의 관계)

  • Lee, Ji Seok;Cho, Jin Hoon;Kim, Ki Uk;Park, Hye-Kyung;Kim, Yun Seong;Lee, Ho Seok;Kim, Yeong Dae;Jeon, Doo Soo;Park, Seung Kyu;Lee, Min Ki;Park, Soon Kew
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.6
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    • pp.492-498
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    • 2007
  • Background: Several lines of evidence suggest that a host's genetic factors influence the outcome of exposure to Mycobacterium tuberculosis. The aim of this study was to determine whether polymorphism in NRAMP1 (natural resistance associated macrophage protein 1) gene is associated with the susceptibility or resistance to tuberculosis infection for patients with drug-sensitive pulmonary tuberculosis (DS-TB) and multi-drug resistant pulmonary tuberculosis (MDR-TB). Methods: Eight genetic polymorphisms of the NRAMP1 gene were investigated in patients suffering with DS-TB (n=100) or MDR-TB (n=102), and in healthy normal controls (NC, n=96). The genetic polymorphisms of NRAMP1 were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequency of D543N A/G heterogygotes was significantly higher in the DS-TB subjects than the NCs (OR=2.10, 95% CI: 1.00 to 4.41, p=0.049). The frequency of 823C/T T/C heterozygotes was significantly higher in the DS-TB subjects (OR=2.79, 95% CI: 1.11 to 7.04, p=0.029) and the MDR-TB subject (OR=3.30, 95% CI 1.33 to 8.18, p=0.010) than in the NCs. However, the frequency of these genotypes was not different between the DS-TB and MDR-TB subjects. Conclusion: A significant association was found between NRAMP1 823 C/T polymorphism and pulmonary tuberculosis. This result suggests that NRAMP1 polymorphism may be involved in the development of pulmonary tuberculosis in Koreans.

Pressure Drop Predictions Using Multiple Regression Model in Pulse Jet Type Bag Filter Without Venturi (다중회귀모형을 이용한 벤츄리가 없는 충격기류식 여과집진장치 압력손실 예측)

  • Suh, Jeong-Min;Park, Jeong-Ho;Cho, Jae-Hwan;Jin, Kyung-Ho;Jung, Moon-Sub;Yi, Pyong-In;Hong, Sung-Chul;Sivakumar, S.;Choi, Kum-Chan
    • Journal of Environmental Science International
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    • v.23 no.12
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    • pp.2045-2056
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    • 2014
  • In this study, pressure drop was measured in the pulse jet bag filter without venturi on which 16 numbers of filter bags (Ø$140{\times}850{\ell}$) are installed according to operation condition(filtration velocity, inlet dust concentration, pulse pressure, and pulse interval) using coke dust from steel mill. The obtained 180 pressure drop test data were used to predict pressure drop with multiple regression model so that pressure drop data can be used for effective operation condition and as basic data for economical design. The prediction results showed that when filtration velocity was increased by 1%, pressure drop was increased by 2.2% which indicated that filtration velocity among operation condition was attributed on the pressure drop the most. Pressure was dropped by 1.53% when pulse pressure was increased by 1% which also confirmed that pulse pressure was the major factor affecting on the pressure drop next to filtration velocity. Meanwhile, pressure drops were found increased by 0.3% and 0.37%, respectively when inlet dust concentration and pulse interval were increased by 1% implying that the effects of inlet dust concentration and pulse interval were less as compared with those changes of filtration velocity and pulse pressure. Therefore, the larger effect on the pressure drop the pulse jet bag filter was found in the order of filtration velocity($V_f$), pulse pressure($P_p$), inlet dust concentration($C_i$), pulse interval($P_i$). Also, the prediction result of filtration velocity, inlet dust concentration, pulse pressure, and pulse interval which showed the largest effect on the pressure drop indicated that stable operation can be executed with filtration velocity less than 1.5 m/min and inlet dust concentration less than $4g/m^3$. However, it was regarded that pulse pressure and pulse interval need to be adjusted when inlet dust concentration is higher than $4g/m^3$. When filtration velocity and pulse pressure were examined, operation was possible regardless of changes in pulse pressure if filtration velocity was at 1.5 m/min. If filtration velocity was increased to 2 m/min. operation would be possible only when pulse pressure was set at higher than $5.8kgf/cm^2$. Also, the prediction result of pressure drop with filtration velocity and pulse interval showed that operation with pulse interval less than 50 sec. should be carried out under filtration velocity at 1.5 m/min. While, pulse interval should be set at lower than 11 sec. if filtration velocity was set at 2 m/min. Under the conditions of filtration velocity lower than 1 m/min and high pulse pressure higher than $7kgf/cm^2$, though pressure drop would be less, in this case, economic feasibility would be low due to increased in installation and operation cost since scale of dust collection equipment becomes larger and life of filtration bag becomes shortened due to high pulse pressure.

Optimization of Biotransformation Process for Sodium Gluconate Production by Aspergillus niger (Aspergillus niger를 이용한 글루콘산 나트륨 생산 생변환 공정의 최적화)

  • 박부수;조병관;이상윤;임승환;김동일;김병기
    • KSBB Journal
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    • v.14 no.3
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    • pp.309-314
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    • 1999
  • In order to produce high concentration of sodium gluconate, optimization of the fermentation conditions, such as glucose concentration, inoculum size, dissolved oxygen concentration and glucose feeding method, was examined. When the glucose concentration was maintained in the range of 30∼50 g/L during the batch fermentation, glucose conversion yield and productivity were 92.2% and 6.0 g/L/hr, respectively. In the case of the low concentration below 30 g/L, the yield decreased by about 25%. As the inoculum size increased above 20%(w/v), lag phase was shortened but the productivity decreased. The dissolved oxygen level of 60∼70% was shown to be the threshold point for 75% of increase in the productivity of sodium gluconate. Finally, optimal glucose feeding rate was determined using various feeding methods such as exponential feeding, feeding based on the average glucose consumption rate and was determined using various feeding methods such as exponential feeding, feeding based on the average glucose consumption rate and on the oxygen uptake rate and etc. Our result shows that glucose feeding, based on the oxygen uptake rate is a very simple, efficient and robust method, especially when oxygen is consumed as a substrate for the bioconversion. Using the above glucose feeding strategy under the optimized condition, 255 g/L of sodium gluconate concentration, 12 g/L/hr of productivity and 95% of glucose conversion yield were achieved with A. niger ACM53.

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Antioxidant, Physiological Activities, and Acetylcholinesterase Inhibitory Activity of Portulaca oleracea Extracts with Different Extraction Methods (추출방법에 따른 쇠비름의 항산화, 생리활성 및 Acetylcholinesterase 저해활성)

  • Kwon, Yu-Ri;Cho, Sung-Mook;Hwang, Seung-Pil;Kwon, Gi-Man;Kim, Jae-Won;Youn, Kwang-Sup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.389-396
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    • 2014
  • The physiological properties of 70% ethanol extracts from Portulaca oleracea with different extraction methods (reflux extraction, RE; autoclave extraction, AE; low temperature high pressure extraction, LTPE) were investigated. The freeze-dried powder yields of RE, AE, and LTPE were 33.78%, 30.80%, and 11.05%, respectively. The color values of L and b were higher in LTPE, and the chroma values were higher in AE and LTPE compared to RE. The total polyphenolics and proanthocyanidin contents in LTPE were significantly higher than in other extracts. The amount of substances related to flavonoids contents was highest in RE (4.30 mg/g), followed by AE (4.06 mg/g), and LTPE (4.00 mg/g). DPPH radical scavenging ability with a concentration of 500 mg% (w/v) were in the following order; LTPE (88.87%)> RE (83.84%)> AE (80.67%). Further, the reducing power, ABTS radical scavenging ability, and nitrite scavenging activity was observed in the same tendency as seen with the DPPH radical scavenging ability. However, the ferrous ion chelating activity of RE (85.45%) and AE (83.88%) was significantly higher than that of LTPE (75.60%). ${\alpha}$-Glucosidase inhibitory activities of RE and LTPE with a concentration of 100 mg% were significantly higher than AE. Xanthine oxidase, and acetylcholinesterase inhibitory activities of LTPE were higher than the other extracts. These results suggest that the extracts from Portulaca oleracea have the potential to act as functional materials, and components of Portulaca oleracea could be effective in the prevention of Alzheimer's disease, and may be used to develop various functional food products.

The Development of Air-kerma Strength Calculation Algorithm in Terms of the Absorbed Dose to Water for HDR Ir-192 Source (기준점에서의 물 흡수선량을 이용한 Ir-192 선원의 공기커마 세기 계산을 위한 알고리즘 개발)

  • Huh, Hyun-Do;Kim, Woo-Chul;Loh, John-Jk;Lee, Suk;Lee, Sang-Hoon;Cho, Sam-Ju;Shin, Dong-Oh;Choi, Jin-Ho;Kwon, Soo-Il;Kim, Seong-Hoon
    • Progress in Medical Physics
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    • v.17 no.4
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    • pp.232-237
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    • 2006
  • The aim of this study was to develop the calculation algorithm of source strength of Ir-192 source In terms of the absorbed dose to water instead of an apparent activity (Ci). For this work the Multi Purpose Brachytherapy Phantom(MPBP) was developed, which was designed to locate the source and the chamber precisely at a specific position Inside the water phantom. The reference point of measurement was set at the 5 cm distance along the transverse axis of the source. For a brachytherapy source calibration, the absorbed dose to water calibration factor ($N_{D.W.Q}$) of an lonization chamber were determined and then apply standard protocols of absorbed dose to water. The calibration factor ($N_{D.W.Q}$) of the ion chamber (TM30013, PTW, Germany) was determined using the EGSnrcCPP Monte Carlo Code. The calculated calibration factor ($N_{D.W.Q}$) was 5.28 cGy/nC. The calculated factor was then used to determine the absorbed dose to water from which the air kerma strength for an Ir-192 source can be easily derived at the reference point (5 cm). The calculated air kerma strength showed discrepancies of -0.6% to +1.8% relative to the air kerma strength provided by the vendor, In this work we demonstrated that the air kerma strength ($S_k$) could be determined from the absorbed dose to water calibration factor for Ir-192 source. In audition, this source calibration method could be applied directly to the dose Calculation formalism of AAPM report TG-43.

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Serum Carbohydrate Antigen 19-9 as an Indicator of Liver Metastasis in Colorectal Carcinoma Cases

  • Dong, Hang;Tang, Jie;Li, Long-Hao;Ge, Jun;Chen, Xin;Ding, Jing;Men, Hai-Tao;Luo, Wu-Xia;Du, Yang;Li, Cong;Zhao, Feng;Chen, Ye;Cheng, Ke;Liu, Ji-Yan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.909-913
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    • 2013
  • Purpose: The liver is the organ to which colorectal carcinomas (CRCs) most commonly metastasize, and surgical resection has been established as the most effective and potentially curative treatment for CRC with liver metastasis (LM). Therefore, surveillance of LM is vital for improvement of prognosis of CRC patients. In this study, we aimed to explore the potential value of carbohydrate antigen 19-9 (CA 19-9), carcinoembryonic antigen (CEA), and marker enzymes in indicating LM with CRC. Methods: Three groups of eligible patients with metastatic cancers were retrospectively included: CRC patients with LM (CRC-LM) or without LM (CRC-NLM), and non-CRC patients with LM (NCRC-LM). All metastatic lesions were identified by CT or MRI. Data on characteristics of the patients, the primary site, the locations of metastasis, CA 19-9, CEA, and biochemical parameters were collected for analysis. Results: A total of 493 patients were retrospectively included. More alcohol consumption was found in CRC-LM than CRC-NLM. Some biochemical enzymes were found to be significantly higher in groups with LM than without (CRC-LM or NCRC-LM v.s CRC-NLM). Both CEA and CA 19-9 were much higher in CRC-LM than CRC-NLM or NCRC-LM. For CRC patients, CA 19-9, ${\gamma}$-glutamyl transpeptidase, CEA and alcohol consumption were identified as independent factors associated with LM. Conclusion: Our analysis suggested the CA 19-9 might be a potential valuable indicator for LM of CRC in the clinic.

The Acceptance Testing of 5 Mega Pixels Primary Electronic Display Devices and the Study of Quality Control Guideline Suitable for Domestic Circumstance (5 Mega 화소 진단용 전자표시장치 인수검사 및 국내 실정에 적합한 정도관리 가이드라인 연구)

  • Jung, Hai-Jo;Kim, Hee-Joung;Kim, Sung-Kyu
    • Progress in Medical Physics
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    • v.18 no.2
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    • pp.98-106
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    • 2007
  • In June 2005, Yonsei University Medical Center, Severance Hospital upgraded a full-PACS system by adding twenty (5 mega pixels) Totoku ME511L flat panel LCD display devices for diagnostic interpretation purposes. Here we report upon the quantitative (or visual) acceptance testing of the twenty Totoku ME511L display devices for reflection, luminance response, luminance spatial dependency, resolution, noise, veiling glare, and display chromaticity based on AAPM TG 18 report. The tools used in the tests included a telescopic photometer, which was used as a colorimeter, illuminance meter, light sources for reflection assessment, light-blocking devices, and digital TG18 test patterns. For selected 8 flat panel displays, mean diffuse reflection coefficient ($R_d$) was $0.019{\pm}0.02sr^{-1}$. In the luminance response test, luminance ratio (LR), maximum luminance difference ($L_{max}$), and deviation of contrast response were $550{\pm}100,\;2.0{\pm}1.9%\;and\;5.8{\pm}1.8%$, respectively. In the luminance uniformity test, maximum luminance deviation was $14.3{\pm}5.5%$ for the 10% luminance of the TG18-UNL10 test pattern. In the resolution test with luminance measurement method, percent luminance (${\Dalta}L$) at the center was $0.94{\pm}0.64%$. In all cases of noise testing, rectangular target In every square in the three quadrants was visible and all 15 targets except the smallest one in the every corner pattern and the center pattern. The glare ratio (GR) was $12,346{\pm}1,995$. The color uniformity, (u',v'), was $0.0025{\pm}0.0008$. Also, the research results of qualify control guideline of primary disply devices suitable for domestic circumstance are presented All test results are in-line with the criteria recommended by AAPM TG18 report and are thus fully acceptable for diagnostic image interpretation. As a result, the acceptance testing schedule described provides not only an acceptance standard but also guidelines for quality control, optimized viewing conditions, and a means for determining the upgrading time of LCD display devices for diagnostic interpretation.

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