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Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation

  • Choi, Jae-Woong;Kim, Jong-Wook;Nguyen, Lap P.;Nguyen, Huu C.;Park, Eun-Mee;Choi, Dong Hwa;Han, Kang Min;Kang, Sang Min;Tark, Dongseob;Lim, Yun-Sook;Hwang, Soon B.
    • Molecules and Cells
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    • v.43 no.5
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    • pp.469-478
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    • 2020
  • Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an important role in the regulation of cell proliferation, migration, and protein-protein interactions. Alteration of LASP-1 expression has been implicated in hepatocellular carcinoma. However, the functional involvement of LASP-1 in HCV propagation and HCV-induced pathogenesis has not been elucidated. Here, we first verified the protein interaction of NS5A and LASP-1 by both in vitro pulldown and coimmunoprecipitation assays. We further showed that NS5A and LASP-1 were colocalized in the cytoplasm of HCV infected cells. NS5A interacted with LASP-1 through the proline motif in domain I of NS5A and the tryptophan residue in the SH3 domain of LASP-1. Knockdown of LASP1 increased HCV replication in both HCV-infected cells and HCV subgenomic replicon cells. LASP-1 negatively regulated viral propagation and thereby overexpression of LASP-1 decreased HCV replication. Moreover, HCV propagation was decreased by wild-type LASP-1 but not by an NS5A binding-defective mutant of LASP-1. We further demonstrated that LASP-1 was involved in the replication stage of the HCV life cycle. Importantly, LASP-1 expression levels were increased in persistently infected cells with HCV. These data suggest that HCV modulates LASP-1 via NS5A in order to regulate virion levels and maintain a persistent infection.

Resumption of Ovarian Cyclicity after Superovulation Treatment to Donor Cow in Hanwoo (한우 공란우 과배란 처리 후 난소 주기 재개에 관한 연구)

  • Son, Jun-Kyu;Choe, Chang-Yong;Cho, Sang-Rae;Yeon, Seong-Heum;Choi, Sun-Ho;Kim, Nam-Tae;Jung, Jin-Woo;Kim, Sung-Jae;Jung, Yeon-Sub;Bok, Nan-Hee;You, Yong-Hee;Son, Dong-Soo
    • Journal of Embryo Transfer
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    • v.25 no.3
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    • pp.149-154
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    • 2010
  • The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100\;{\mu}g$ GnRH at the time of Ind insemination. Embryos were recovered 7 or 8 days after the 1st insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. 50.0 percentage of the cows (18/36) had normal resumption of ovarian cyclicity (resumption within 40 days after superovulation), and 50.0% (18/36) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II (first ovulation did not occur until $\geq$ 40 days after superovulation, i.e. delayed first ovulation 33.3%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and 10$\leq$ of corpora lutea (CL) was 7.3 and 13.9, respectively. The number of transferable embryos differed between < 10 and 10$\leq$ CL was 4.2 and 5.1, respectively. 11.1 percentage of the cows (4/36) did not resumption their ovarian cyclicity until 60 days after superovulation treatment.

Technical Feasibility of Ethanol-Kerosene Blends for Farm Kerosene Engines (에타놀-석유(石油) 혼합연료(混合燃料)의 농용석유(農用石油)엔진에의 이용(利用)에 관(關)한 연구(硏究))

  • Bae, Yeong Hwan;Ryu, Kwan Hee
    • Journal of Biosystems Engineering
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    • v.7 no.1
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    • pp.53-61
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    • 1982
  • As an attempt to reduce the consumption of petroleum resources and to improve the performance of a kerosene engine, a series of experiments was conducted using several kinds of ethanol-kerosene blends under the various compression ratios. The engine used in this study was a single-cylinder, four-cycle kerosene engine having a compression ratio of 4.5. To investigate the feasibility of ethanol-kerosene blends in the original engine, kerosene and blends of 5-percent, 10-percent, and 20-percent-ethanol, by volume, with kerosene were used. And to investigate the feasibility of improving the performance of the kerosene engine, a portion of the cylinder head was cut off to increase the compression ratio up to 5.0 by reducing the combustion chamber volume. Kerosene and blends of 30-percent and 40-percent-ethanol, by volume, with kerosene were used for the modified engine with an increased compression ratio. Variable speed tests at wide-open throttle were also conducted at five speed levels in the range of 1000 to 2200 rpm for each compression ratio and fuel type. Volumetric efficiency, engine torque, and brake specific fuel consumption were determined, and brake thermal efficiency based on the lower heating values of kerosene and ethanol was calculated. The results obtained in the study are summarized as follows: A. Test with the original engine: (1) No abnormal conditions were found when burning ethanol-kerosene blends in the original engine. (2) Volumetric efficiency increased with ethanol concentration in blends. When burning blends of 5-percent, 10-percent, and 20-percent ethanol, by volume, with kerosene, average volumetric efficiency increased 1.6 percent, 2.6 percent, and 4.1 percent respectively, than when burning kerosene. (3) Mean engine torque increased 5.2 percent for 5-percent-ethanol blend, 9.3 percent for 10-percent-ethanol blend, and 11.5 percent for 20-percent-ethanol blend than for kerosene. Increase in engine torque when using ethanol-kerosene blends was due to the improved combustion characteristics of ethanol as well as an increase in volumetric efficiency. (4) Up to ethanol concentration of 20 percent, mean brake specific fuel consumption was nearly constant inspite of the difference in heating value between ethanol and kerosene. (5) Brake thermal efficiency increased 0.3 percent for 5-percent-ethanol blend, 3.8 percent for 10-percent-ethanol blend, and 6.8 percent for 20-percent-ethanol blend than for kerosene. B. Test with the modified engine with an increased compression ratio: (1) When burning kerosene, mean volumetric efficiency, engine torque, and brake thermal efficiency were somewhat lower than for the original engine. (2) Engine torque increased 15.1 percent for 30-percent-ethanol blend and 18.4 percent for 40-percent-ethanol blend than for kerosene. (3) There was no significant difference in brake specific fuel consumption regardless of ethanol concentration in blends. (4) Brake thermal efficiency increased 15.0 percent for 30-percent-ethanol blend and 19. 5 percent for 40-percent-ethanol blend than for kerosene.

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Relationship between the Expression of Forkhead box M1 (FoxM1) and $p27^{kip1}$ in Non-Small Cell Lung Cancers

  • Lee, Kyung-Eun;Hong, Young-Seoub;Choi, Phil-Jo;Um, Soo-Jung;Son, Choon-Hee;Roh, Mee-Sook
    • Biomedical Science Letters
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    • v.14 no.4
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    • pp.243-248
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    • 2008
  • The Forkhead box M1 (FoxM1) has been shown to regulate transcription of cell cycle genes essential for $G_1$-S and $G_2$-M progression, including $p27^{kip1}$. The $p27^{kip1}$ gene is a member of the universal cyclin-dependent kinase inhibitor family. Immunohistochemical studies for FoxM1 and $p27^{kip1}$ were performed in 154 lung cancers (69 squamous cell carcinomas (SCC) and 85 adenocarcinomas (ADC)). Immunoreactivity for FoxM1 and $p27^{kip1}$ were found in 79 (51.3%) and 49 (31.8%) out of 154 cases, respectively. Forty-six (58.2%) of the 79 cases with a positive FoxM1 immunoreactivity showed a negative $p27^{kip1}$ expression in 154 lung cancers. According to histologic type, 22 (53.7%) of the 41 SCC cases with a positive FoxM1 immunoreactivity showed a negative $p27^{kip1}$ expression and 24 (63.2%) of the 38 ADC cases with a positive FoxM1 immunoreactivity showed a negative $p27^{kip1}$ expression. The expression of $p27^{kip1}$ was significantly higher in the SCC than in the ADC (P=0.050). There were no significant associations between the FoxM1 and $p27^{kip1}$ expressions and other clinicopathologic factors. These findings suggest that FoxM1 overexpression may diminish the expression of $p27^{kip1}$ protein in lung cancers. Further studies are needed to define the relation between FoxM1 and $p27^{kip1}$ for examining the mechanisms of tissue-specific FoxM1 expression.

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A Study on the Thermo-mechanical Characteristics and Adhesion Reliability of Anisotropic Conductive Films Depend on the Curing Methods of Epoxy Resins (에폭시 레진의 경화방법에 따른 이방성 전도필름의 접합신뢰성 및 열적기계적 특성 변화)

  • Gil, Man-Seok;Seo, Kyoung-Won;Kim, Jae-Han;Lee, Jong-Won;Jang, Eun-Hee;Jeong, Do-Yeon;Kim, Su-Ja;Kim, Jeong-Soo
    • Polymer(Korea)
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    • v.34 no.3
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    • pp.191-197
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    • 2010
  • To improve the curing method of anisotropic conductive film (ACF) at low temperature, it was studied to replace the thermal latent curing agent of imidazole compounds by the curing agent of cationically initiating type. Thermo-mechanical properties such as glass transition temperature, storage modulus, and coefficient of thermal expansion were investigated for the analysis of curing behavior. The reliability of ACF were observed in thermal cycle and high temperature-high humidity test. ACF using cationic initiator showed faster curing, lower CTE, and higher $T_g$ than the case of using imidazole curing agent, which is important for the high temperature stability. Furthermore, ACF using cationic initiator maintained a stable contact resistance in reliability test, although it was cured at low temperature and fast rate. With these results, it was confirmed that the curing method of epoxy had great effect on thermo-mechanical properties and reliability of ACF.

Influence of the Soluble Salt on the Exfoliation of the Stone Monument (수용성염이 석조문화재 표면 박리현상에 미치는 영향)

  • Do, Jin-Young;Lim, Kwon-Woong
    • Journal of Conservation Science
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    • v.22
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    • pp.121-134
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    • 2008
  • The mechanism of stone exfoliation and its cause in relation to chemical weathering by soluble salt were studied. Chemical, mineralogical and physical analysis was performed in exfoliation samples from stone monuments. The representative salt is gypsum in the exfoliation samples. In order to understand the salt reaction, stone samples(tuff and granite) were treated with two type of the salt, gypsum and sodium sulfate, which have different solubility. The capillary water uptakes are slight increased in impregnated with Na2SO4 and weathering simulation of two rock types. It means that the rock is deteriorated in the near of the surface by $Na_2SO_4$. $CaSO_4{\cdot}2H_2O$ bring out the thicker exfoliation than $Na_2SO_4$ because it is penetrated into the deeper zone and amount of accumulated salt is more abundant in the inner part than in the near of the surface. The exfoliation was formed in the tuff by salt treatment and 30cycle of weathering simulation, but there are not significant symptoms of exfoliation in the granite by same condition. This result was caused by the different capillary water uptakes and porosity of the rocks. In the tuff, salt solutions are penetrated into the inner part due to its high capillary water uptakes and porosity but the granite, which has low value relatively, can be formed thinner exfoliation.

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Performances of the Used Frying Oil on a Small Diesel Engine (폐식용유를 이용한 소형 디젤기관의 성능)

  • 김성태;정형길;김영복
    • Journal of Biosystems Engineering
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    • v.26 no.3
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    • pp.209-220
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    • 2001
  • This study was carried out to investigate the usability of the used frying oil, which was extracted from soybean, as one of the alternative fuel of a small diesel engine. For the experiment, NO. 2 diesel oil [D], used frying oil [UF], and their volumetric blends were applied and analysis of the properties and compositions of the experimental fuels were conducted. A four cycle diesel engine with single cylinder, water cooling system, maximum output 8.1 ㎾/2,200 rpm was selected and a direct injection chamber and a precombustion chamber were attached alternately. The results obtained were as follows: 1. Engine power (BHP) were increased from 4.13~4.27㎾ to 9.08~9.15㎾ for diesel oil, from 4.05~4.19㎾ to 8.44~8.92㎾ for UF, and from 4.01~4.48㎾ to 8.69~9.16㎾ for blend fuel, as the engine speed increased from 1,000 rpm to 2,200 rpm. The BHP in case of the direct combustion chamber were fluctuated higher than those of the pre-combustion chamber. 2. With the engine speed increased, torque of the engine were increased from 39.50~40.80 N.m to 42.89 N.m, then decreased to 39.44~39.77 N.m for diesel oil, and increased from 38.73~40.04 N.m to 40.12~40.82 N.m then decreased as 36.53~38.76 N.m for UF. Torque of the blend fuels were increased from 38.75~41.76 N.m to 40.47~42.89 N.m then decreased to 37.73~39.78 N.m. There is no significant difference of torque between the type of combustion chambers. 3. The specific fuel consumption of the UF was increased about 20 percent depending on the engine speed variations. And in case of direct injection chamber, about 12 percent lower fuel consumption was observed than that of precombustion chamber. 4. NOx emission of the UF was higher than that of diesel oil at above 1,800rpm of the engine speed. In case of the direct injection chamber, NOx emission was revealed higher about 59 percent than that of the precombustion chamber, depending on the range of the engine speeds. 5. Smoke emission was decreased in case of UF compared with diesel oil on direct injection chamber. When using precombustion chamber smoke emission was a little higher than that of the direct injection chamber were showed at the engine speed range. 6. At all the engine speed range, exhaust gas temperatures were decreased 2~3$^{\circ}C$ for UF used engine compared with those of the diesel oil. The exhaust gas temperature of the direct injection chamber was higher than that of the precombustion chamber by 72$^{\circ}C$. 7. Unburnt materials remained in the cylinder in case of the pre-combustion chamber was smaller and softer than that of the direct combustion chamber. 8. The feasibility of the blend fuel B-1 and B-2 were verified as a direct combustion chamber was attached to the diesel engine, with respect to the power performance of the engine.

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A Study on Electrolysis of Heavy Water and Interaction of Hydrogen with Lattice Defects in Palladium Electrodes (팔라디움전극에서 중수소의 전기분해와 수소와 격자결함의 반응에 관한 연구)

  • Ko, Won-Il;Yoon, Young-Ku;Park, Yong-Ki
    • Nuclear Engineering and Technology
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    • v.24 no.2
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    • pp.141-153
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    • 1992
  • Excess tritium analysis was peformed to verify whether or not cold fusion occurs during electrolysis of heavy water in the current density range of 83~600 mA/$\textrm{cm}^2$ for a period of 24 ~ 48 hours with use of palladium electrodes of seven different processing treatments and geometries. The extent of recombination of D$_2$ and $O_2$gases in the electrolytic cell was measured for the calculation of accurate enthaplpy values. The behavior and interaction of hydrogen atoms with defects in Pd electrodes were examined using the Sieverts gas charging and the positron annihilation(PA) method. Slight enrichment of tritium observed was attributed to electrolytic enrichment but not to the formation of a by-product of cold fusion. The extent of recombination of D$_2$and $O_2$gases was 32%. Hence the excess heat measured during the electrolysis was considered to be due to the exothermic reaction of recombination but not to nuclear fusion. Lifetime results from the PA measurements on the Pd electrodes indicated that hydrogen atoms could be trapped at dislocations and vacancies in the electrodes and that dislocations were slightly more preferred sites than vacancies. It was also inferred from R parameters that the formation of hydrides was accompanied by generation of mostly dislocations. Doppler broadening results of the Pd electrodes indicated that lattiec defect sites where positrons were trapped first increased and then decreased, and this cycle was repeated as electrolysis continued. It can be inferred from PA measurements on the cold-rolled Pd and the isochronally annealed Pd hydride specimens that microvoid-type defects existed in the hydrogen-charged electrode specimen.

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Comparison of quality of 30:2 vs. 2:30 CPR in manikins (심폐소생술 방법 변화에 따른 quality 비교 - 30:2와 2:30 비교분석실험 -)

  • Uhm, Tai-Hwan;Yoou, Soon-Kyu;Choi, Hea-Kyung;Jung, Ji-Yeon
    • The Korean Journal of Emergency Medical Services
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    • v.14 no.3
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    • pp.71-81
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    • 2010
  • Purpose: To minimize an interruption in chest compression, reduce the hands-off time, the American Heart Association has recommended the ratio of chest compression to ventilation ratio to 30:2 from 2005 CPR guideline to 2010 CPR guideline. However, current studies have shown that the hands-off time was > 10 seconds with that method. For this reason, we devised new CPR method that a ventilation to chest compression ratio of 2:30 to reduce pt assessment time and skipped the assessment step of carotid artery pulse would be a more effective way to reduce the hands-off time & the time to set the CPR. According to the more detailed purpose are listed below. 1) We would like to confirm efficiency of a ventilation to chest compression ratio of 2:30 than a chest compression to ventilation ratio of 30:2 to reduce the hands-off time & the time to set the CPR. 2) We would like to evaluate possibility of increasing for chest compression accuracy of a ventilation to chest compression ratio of 2:30 than a chest compression to ventilation ratio of 30:2 3) We would like to evaluate possibility of increasing for ventilation accuracy of a ventilation to chest compression ratio of 2:30 than a chest compression to ventilation ratio of 30:2 Methods: According to 2005 American Heart Association Guidelines, 60 paramedic students(20 students X freshmen, sophomore, junior) performed 5 cycles of 3~ chest compressions : 2 ventilations after A, B, C evaluation with Laerdal Resusci R Anne SkillReporters. After 5 minutes rest, the 60 students performed 5 cycles of 2 ventilations : 30 chest compressions after A, B evaluation with the manikins between 13 and 17 September 2010. The short reports including speed & accuracy of chest compression, respiratory, CPR cycle were gained from the manikins. Hands-off times were measured by assistants. Results: Recently, the importance of high quality CPR was emphasized in order to perform the CPR faster and more accurate. To find out improving the conventional CPR method, we switch the procedure of the compression and the ventilation. By switching the procedure back and forth, we are able to compare the effectiveness of CPR between two type of CPR method which are 2:30 and 30:2 methods. 2:30 is that the breaths is delivered twice, first and perform 30 compressions while 30:2 perform 30 compressions first and give 2 breaths followed by the ABC method. Also, we verify the effectiveness of the hands off time, compression accuracy of the compression through the comparison of the two procedure as mentioned earlier. Consequently research verified that 2:30 is the efficient by providing faster set up delivering more accurate chest compression. Conclusion: 2:30 can minimize a time delay from cardiac standstill until starting the chest compression. In addition, hands-off time which is an interruption in chest compression can be shortened by 2:30 method, which result to effective oxygenation of coronary artery & maintenance of the bloodstream. Once again, performing the 2:30 method provide lessen hands off time and increase the accuracy of the chest compression.

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Study on ICT convergence in Lentinula edodes (Shiitake) cultivation system using Automated container (컨테이너형 수출용 버섯식물공장시스템설계 및 표고버섯 생산 연구)

  • Jo, Woo-Sik;Lee, Sung-Hak;Park, Woo-Ram;Shin, Seung-Ho;Park, Chang-Min;Oh, Ji-Hyun;Park, Who-Won
    • Journal of Mushroom
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    • v.15 no.4
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    • pp.264-268
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    • 2017
  • In the 21st century, information and communication technology (ICT) worldwide presents a new vision for agriculture. Time and place, as well as the high-tech industry, to overcome barriers to the fusion of the so-called "smart agriculture," are changing the agricultural landscape. Core container production in precision agriculture for mushroom cultivation, optimal temperature, humidity, irradiation, self-regulation of factors such as carbon dioxide, and environment for mushroom cultivation were adopted. Lentinula edodes (shiitake) is an edible mushroom native to East Asia, cultivated and consumed in many Asian countries. It is considered to be medicinal in certain practices of traditional medicine. We used different controlled light sources (Blue-Red-White-combined LED, blue LED, red LED, and fluorescent light) with different LED radiation intensities (1.5, 10.5, and $20.5{\mu}mol/m^2s$ for LEDs) to compare growth and development. Mushrooms were treated with light in a 12-hour-on/12-hour-off cycle, and maintained in a controlled room at $19{\sim}21^{\circ}C$, with 80~90% humidity, and an atmospheric $CO_2$ concentration of 1,000 ppm for 30 days. Growth and development differed with the LED source color and LED radiation intensity. Growth and development were the highest at $10.5{\mu}mol/m^2s$ of blue LED light. After harvesting the fruit bodies, we measured their weight and length, thickness of pileus and stipe, chromaticity, and hardness. The $10.5{\mu}mol/m^2s$ blue-LED-irradiated group showed the best harvest results with an average individual weight of 39.82 g and length of 64.03 mm, pileus thickness of 30.85 mm and pileus length of 43.22 mm, and stipe thickness of 16.96 mm with fine chromaticity and hardness. These results showed that blue LED light at $10.5{\mu}mol/m^2s$ s exerted the best effect on the growth and development of L. edodes (shiitake) mushroom in the ICT-system container-type environment.