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In vitro Development of Somatic Cell Nuclear Transferred Bovine Embryos Following Activation Timing in Enucleated and Cryopreserved MII Oocytes (탈핵 후 동결한 MII 난자의 활성화 시기가 체세포 핵치환 이후 소 난자의 체외발달에 미치는 영향)

  • 박세필;김은영;김선균;이영재;길광수;박세영;윤지연;이창현;정길생
    • Korean Journal of Animal Reproduction
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    • v.26 no.3
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    • pp.245-252
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    • 2002
  • This study was to evaluate the in vitro survival of bovine enucleated MII (eMII) oocytes according to minimum volume cooling (MVC) freezing method and activation timing, and their in vitro development after somatic cell nuclear transfer (SONT). in vitro matured bovine oocytes for 20 h were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst, and their 1st polar body and MII plate were removed by enucleation micropipette under UV filter. Also, eMII oocytes were subjected to activation after (group II) and before (group III) vitrification in 5 ${\mu}{\textrm}{m}$ ionomycin added CRlaa medium for 5 min. For vitrification, eMll oocytes were pretreated with EG10 for 5 min, exposed to EG30 for 30 sec and then directly plunged into L$N_2$. Thawing was taken by 4-step procedures at 37$^{\circ}C$. Survived eMII oocytes were subjected to SONT with cultured adult bovine ear cells. Reconstructed oocytes were cultured in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide and 2.5 $\mu\textrm{g}$/$m\ell$ of cytochalasin D added CRlaa medium for 1 h, and then in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide added CRlaa medium for 4 h. Subsequently, the reconstructed oocytes were incubated for 2 days and cleaved embryos were further cultured on cumulus-cell monolayer drop in CRlaa medium for 6 days. Survival rates of bovine vitrified-thawed eMII oocytes in group II (activation after vitrification and thawing) and III (activation before vitrification) were 81.0% and 84.9%, respectively. Fusion rates of cytoplasts and oocytes in group II and III were 69.0% and 70.0%, respectively, and their results were not different with non-frozen NT group (control, 75.2%). Although their cleaved rates (53.4% and 58.4%) were not different, cytoplasmic fragment rate in group II (32.8%) was significantly higher than that in group III (15.6%)(P<0.05). Also, subsequent development rate into >morula in group II (8.6%) was low than that in group III(15.6%). However, in vitro development rate in group III was not different with that in control (24.8%). This result suggested that MVC method was appropriate freezing method for the bovine eMII oocytes and vitrified eMII oocytes after pre-activation could support in vitro embryonic development after SONT as equally well as fresh oocytes.

Physico-Chemical Properties and Composition of Venison in Deer Breeds. (사슴 품종별 녹육의 이화학적 특성에 관한 연구)

  • 이길왕
    • Journal of Life Science
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    • v.14 no.3
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    • pp.525-530
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    • 2004
  • This study has been performed to examine physico-chemical properties and composition of venison in Deer breeds. According the results, moisture content in the venison ranged between 74% and 75%, and there were no significant differences among the breeds, or among the cutting parts. Fat content in the meats was estimated as less than 1% except in loin, protein contents was estimated as more than 22%, and ash content was estimated as more than 1.5%. For the contents of inorganic material in the loin, Fe$^{3+}$ content was higher in Elk than in other breeds, the contents of $Cu^{2+}$, $Mn^{2+}$, $K^{+}$, $Mg^{2+}$ and $Ca^{2+}$were higher in Red deer than in others, and the contents of $Fe^{3+}$, $K^{+}$, and $P^{+}$ were higher in Sika deer than in others. For the contents of inorganic material in the leg, the contents of $Cu^{2+}$, $Zn^{2+}$, $K^{+}$, $Ca^{2+}$ and $Mg^{2+}$ were higher in Elk than in other breeds, the contents of $Fe^{3+}$ and$P^{+}$ were higher in Red deer than in others, and the contents of $Cu^{2+}$ and $Fe^{3+}$were higher in Sika deer than in others. Amino acid contents in the venison alanine, proline, glycine, lycine and histidine were highly contained. and the contents of asparagine, cysteine, isoleucine and threonine were low. For the differences of amino acid contents among the breeds, the level was the higher in Sika deer than those of others. For the fatty acid in loin, the contents of myristic acid, oleic acid and linolenic acid in Sika deer were higher than those of others, the contents of myristic acid, palmitoleic acid, oleic acid and arachidonic acid in Red deer were higher than those of others and in Elk, the contents of palmitic acid and stearic acid were higher than those of others. For the contents of unsaturated fatty acid and polyunsaturated fatty acids, the level was higher in Red deer than those of others. For the fatty acid in leg, the contents of oleic acid, linolenic acid, linolenic acid and arachidonine acid in Sika deer were higher than those of others, the contents of myristic acid, palmitic acid, palmitoleic acid and oleic acid in Red deer were higher than those of others and in Elk, the contents of stearic acid and linolenic acid were higher than those of others. For the contents of saturated fatty acid, the level was lowest in Sika deer. For the contents of unsaturated fatty acid and polyunsaturated fatty acids were respectively highest in Sika deer. Comparing the contents inorganic materials between loin and leg, the concentrations in leg were generally higher than that of loin.n.

Studies on Dry Matter Yields , Chemical Composition and Net Energy Accumulation in Three Leading Temperate Grass Species II. Synthesis and accumulation pattern of nonstructural carbohydrate (주요 북방형목초의 건물수량 , 화학성분 및 New Energy 축적에 관한 연구 II. 비구조성탄수화물의 합성 및 축적형태)

  • ;;F. Muhlschlegel
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.6 no.2
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    • pp.111-118
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    • 1986
  • Sysnthesis and accumulation pattern or nonstructural carbohydrates in orchardgrass (Dactylis glomerata L.) cv. Potomac and Baraula, perennial ryegrass (Lolium perenne L.) cv. Reveille and Semperweide and meadow fescue (Festuca pratensis Huds.) cv. Cosmos 11 and N.F.G. were studied under different meteorological environments and cutting managements. The field experiments were conducted as a split plot design with three cutting regimes of 6-7 cuts at grzing stage, 4-5 cuts at silage stage and 3 cuts at hay stage in Korea and West Germany from 1975 to 1979. The results obtained are summarized as follows: 1. Accumlation of nonstructural carbohydrates in temperate grasses was influenced by grass species and regional climatic environments. Total nonstructural carbohydrates (TNC) of orchardgrass, perennial ryegrass and meadow fescue in Korea, taken as average of all cutting regimes, were shown a value of 4.39%, 6.08% and 8.01%, respectively, while those under cool summer climatic condition in West Germany accumulated to 10.42% (orchardgrass), 18.02% (perennial ryegrass) and 12.73% (meadow fescue). 2. Nonstructural carbohydrates in orchardgrass were accumulated mainly as mono-and disaccharose, while those in perennial ryegrass resreved as fructosan. The contents of fructosan and mono-and disaccharose were 1.34% and 3.04% for orchardgrass, 3.25% and 2.83% for perenninal ryegrass, respectively. Meadow fescue had a concentration of 3.93% fructosan and 4.08% mono-and disaccharose. 3. Synthesis and accumulation of nonstructural carbohydrates in temperate grasses were negative associated with increasing of air temperature (P$\leq$ 0.1%). Under hot stress during summer season in Korea, the contents of fructosan, mono-and disaccharose were decreased to about 0.34% nd 1.28% from a value of 1.34% and 2.69% in spring season. In Freising and Braunschweig, the concentration of reserved carbohydrates was less influenced by growing season. 4. Synthesis and accumulation pattern of nonstructural carbohydrates were shown a great respons to cutting frequency of the plants. Frequent cutting system under high temperature lowered the accumulation of reserved carbohydrates, especially fructosan and also caused to decrease the plant regrowth. However, under cool temperature, it shows a less differences of tructosan, mono-and disaccharose in the plants at all cutting systems.

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Growth and Physiological Characteristics of Pinus densiflora Seedlings in Response to Open-field Experimental Warming using the Infrared Lamp (적외선등을 이용한 실외 실험적 온난화 처리가 소나무 묘목의 생장과 생리적 특성에 미치는 영향)

  • Lee, Sun Jeoung;Han, Saerom;Yoon, Tae Kyung;Han, Seung Hyun;Jung, Yejee;Yun, Soon Jin;Son, Yowhan
    • Journal of Korean Society of Forest Science
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    • v.102 no.4
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    • pp.522-529
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    • 2013
  • Climate change will affect the physiological traits and growth of forest trees. This study was conducted to investigate the effects of an experimental warming on growth and physiological characteristics of Pinus densiflora S. et Z. seedlings. One-year-old P. densiflora seedlings were planted in control and warmed plots in April 2010. The air temperature of warmed plots was increased by $3^{\circ}C$ using infrared lamps from November 2010. We measured shoot height, root collar diameter, above and below ground biomass, chlorophyll contents and leaf nitrogen concentration from March 2011 to March 2013. Seedling height and root collar diameter showed no significant difference between warmed and control plots except for root collar diameter measured in June 2012. Seedling leaf biomass was lower in the warmed ($23.94{\pm}2.10g$) than in the control ($26.08{\pm}1.72g$) plots in 2013. Shoot to root ratio (S/R ratio) was lower in the warmed ($1.09{\pm}0.07$) than in the control ($1.31{\pm}0.10$) plots in 2013. Leaf nitrogen concentrations and chlorophyll contents were not significantly different between warmed and control plots except for leaf nitrogen concentration in 2011. Leaf C/N ratio was increased in 2012 under the warming treatment. Low growth and S/R ratio in warmed plots might be related to the higher temperature and water stress. In the future, the below-ground carbon allocation of P. densiflora might be increased by global warming due to temperature and water stress.

Risk Factors for Hearing Loss in Very Low Birth Weight Infants: Results of Hearing Test in Infants <1,500 g (극소 저체중 출생아에서 청력 손상에 영향을 미치는 요인: 1,500 g미만의 청력 검사 결과)

  • Sung, Min-Jung;Han, Young-Mi;Park, Kyung-Hee;Lee, Il-Woo;Byun, Shin-Yun
    • Neonatal Medicine
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    • v.18 no.2
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    • pp.328-336
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    • 2011
  • Purpose: An association between very low birth weight infants(VLBWI) and hearing loss has long been recognized. Early identification and intervention for hearing loss benefits language and speech/cognitive development. We investigated the risk factors and clinical outcomes of hearing loss among VLBWI. Methods: We analyzed the results of auditory brainstem response (ABR) testing of VLBWI. These infants were admitted to the neonatal intensive care unit (NICU) of Pusan National University Yangsan Hospital between December 2008 and February 2011. A follow-up was conducted subsequently. Results: ABR evaluations were performed on 65 infants, and 31 showed abnormal results (47.7%). Among the 31 infants, 10 were classified with moderate/severe/profound hearing loss (15.4%). The infants with abnormal ABR had a higher incidence of low birth weight, prolonged ventilator care, cumulative dose of furosemide, and the lowest $PaO_2$ (P<0.05). Those with moderate/severe/profound hearing loss had a higher incidence of low Apgar scores at 5 minutes (odds ratio[OR],0.34; 95% confidence interval[CI],0.13-0.89), prolonged ventilator care (OR,1.06; 95% CI,1.01-1.12), and mild hearing loss compared to those without profound hearing loss. Follow-up evaluations on eight infants with ABR reveled improvements 5.6${\pm}$3.9 months later. One infant, who had profound hearing loss in both ears, used a hearing aid. Conclusion: Factors influencing hearing loss at the first VLBWI hearing screening test included lower Apgar scores at 5 min and prolonged use of a ventilator. Most VLBWI with hearing losses were expected to recover after several months of follow-up.

The Significance of Maturation Score of Brain Magnetic Resonance Imaging in Extremely Low Birth Weight Infant (초극소 저체중 출생아의 뇌 MRI 상 Maturation Score의 의의)

  • Song, In-Gu;Kim, Su-Yeong;Kim, Cur-Rie;Kim, Yoon-Joo;Shin, Seung-Han;Lee, Seung-Hyun;Lee, Jae-Myoung;Lee, Ju-Young;Kim, Ji-Young;Sohn, Jin-A;Lee, Jin-A;Choi, Chang-Won;Kim, Ee-Kyung;Cheon, Jung-Eun;Kim, Woo-Sun;Kim, Han-Suk;Kim, Byeong-II;Kim, In-One;Choi, Jung-Hwan
    • Neonatal Medicine
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    • v.18 no.2
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    • pp.310-319
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    • 2011
  • Purpose: The aim of this study was to investigate the effect of perinatal risk factors on brain maturation and the relationship of brain maturation and neurodevelopmental outcomes with brain maturation scoring system in brain MRI. Methods: ELBWI infants born at the Seoul National University Children's Hospital from January 2006 to December 2010 were included. A retrospective analysis was performed with their medical record and brain MR images acquired at near full term. We read brain MRI and measured maturity with total maturation score (TMS). TMS is a previously developed anatomic scoring system to assess brain maturity. The total maturation score was used to evaluate the four parameters of maturity: (1) myelination, (2) cortical infolding, (3) involution of glial cell migration bands, and (4) presence of germinal matrix tissue. Results: Images from 124 infants were evaluated. Their mean gestational age at birth was 27.1${\pm}$2.1 weeks, and mean birth weight was 781.5${\pm}$143.9 g. The mean TMS was 10.8${\pm}$2.0. TMS was significantly related to the postmenstrual age (PMA) of the infant, increasing with advancing postmenstrual age (P<0.001). TMS showed no significance with neurodevelopmental delay, and with brain injury, respectively. Conclusion: TMS was developed for evaluating brain maturation in conventional brain MRI. The results of this study suggest that TMS was not useful for predicting neurodevelopmental delay, but further studies are needed to make standard score for each PMA and to re-evaluate the relationship between brain maturation and neurodevelopmental delay.

Production and Characterization of Extracellular Polysaccharide Produced by Pseudomonas sp. GP32 (Pseudomonas sp. GP32에 의해 생산된 세포 외 다당류의 생산 및 특성)

  • Lee, Myoung Eun;Lee, Hyun Don;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1027-1035
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    • 2015
  • A strain GP32 which produces a highly viscous extracellular polysaccharide was conducted with soil samples and identified as Pseudomonas species. The culture flask conditions for the production of extracellular polysaccharide by Pseudomonas sp. GP32 were investigated. The most suitable carbon and nitrogen source for extracellular polysaccharide production were galactose and (NH4)2SO4. The optimum carbon/nitrogen ratio for the production of extracellular polysaccharide was around 50. The optimum pH and temperature for extracellular polysaccharide production was 7.5 and 32℃, respectively. In batch fermentation using a jar fermentor, the highest extracellular polysaccharide content (15.7 g/l) was obtained after 70 hr of cultivation. The extracellular polysaccharide produced by Pseudomonas sp. GP32 (designated Biopol32) was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation, and gel permeation chromatography. Biopol32, which has an estimated molecular weight of over 3×107 datons, is a novel polysaccharide derived from sugar components consisting of galactose, glucose, gulcouronic acid and galactouronic acid in an approximate molar ratio of 1.85 : 3.24 : 1.00 : 1.42. The solution of Biopol32 showed non-Newtonian characteristics. The viscosity of Biopol32 exhibited appeared to be higher at all concentration compared to that of zooglan from Zoogloea ramigera. An analysis of the flocculating efficiency of Biopol32 in industry wastewater (food, textile, and paper wastewater) revealed chemical oxygen demand (COD) reduction rates 58.4-67.3% and suspended solid (SS) removal rates 82.6-91.3%. Based on these results, Biopol32 is a possible candidate for industrial applications such as wastewater treatment.

Pro-apoptotic and Anti-adipogenic Effects of Proso Millet (Panicum miliaceum) Grains on 3T3-L1 Preadipocytes (기장(Panicum miliaceum)의 마우스 3T3-L1 세포에 대한 에폽토시스 유발 및 지방세포형성 억제 효능)

  • Jun, Do Youn;Lee, Ji Young;Han, Cho Rong;Kim, Kwan-Pil;Seo, Myung Chul;Nam, Min Hee;Kim, Young Ho
    • Journal of Life Science
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    • v.24 no.5
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    • pp.505-514
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    • 2014
  • To examine the anti-obese activity of miscellaneous cereal grains, 80% ethanol extracts from eight selected miscellaneous cereal grains were compared for their cytotoxic effects on 3T3-L1 murine preadipocytes. The ethanol extract of proso millet exhibited the highest cytotoxicity. Further fractionation of the ethanol extract with methylene chloride, ethyl acetate, and n-butanol showed that the cytotoxicity of the ethanol extract was mainly partitioned into the butanol fraction. As compared with differentiated mature adipocytes, 3T3-L1 preadipocytes were more susceptible to the cyctotoxicity of the butanol fraction. When each organic solvent fraction (25 ${\mu}g/ml$) was added during the differentiation period for 6 days, the cell viability was not affected significantly except for the butanol fraction, but the intracellular lipid accumulation declined to a level of 81.5%~50.3% of the control. The Oil Red O staining data also demonstrated that the ethanol extract as well as the butanol fraction could inhibit the differentiation of 3T3-L1 preadipocytes into mature adipocytes. The presence of the butanol extract during the induced adipocytic differentiation also resulted in a significant reduction in the expression levels of critical adipogenesis mediators $(C/EBP{\alpha}$, $PPAR{\gamma}$, aP2, and LPL) to a barely detectable or undetectable level and the cells retained the fibroblast-like morphology of 3T3-L1. In 3T3-L1 cells, the cytotoxicity of the butanol fraction (50-100 ${\mu}g/ml$) was accompanied by mitochondrial membrane potential (${\Delta}{\psi}m$) loss, caspase-3 activation, and PARP degradation. Taken together, these results indicate that proso millet grains possess pro-apoptotic and anti-adipocytic activities toward adipocytes, which can be applicable to prevention of obesity.

Study on the Optimum Range of Weight-Age Data for Estimation of Growth Curve Parameters of Hanwoo (한우의 체중 성장곡선 모수 추정을 위한 체중 측정 자료의 최적 범위에 관한 연구)

  • Cho, Y.M.;Yoon, H.B.;Park, B.H.;Ahn, B.S.;Jeon, B.S.;Park, Y.I.
    • Journal of Animal Science and Technology
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    • v.44 no.2
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    • pp.165-170
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    • 2002
  • Mature weight (A) and rate of maturing (k) estimated by nonlinear regression were studied to determine the optimum age range over which the estimate of growth curve parameters can be estimated. The weight-age data from 1,133 Hanwoo bulls at Hanwoo Improvement Center of N.A.C.F. were used to fit the growth curve using Gompertz model. All available weight data from birth to the specific age of months were used for the estimation of parameters: the six specific ages used were 12, 14, 16, 18, 20 22 and 24 months of age. The mean estimates of mature weight (A) were 966.5, 1,255.9, 1,126.2, 916.5, 842.2, 780.9 and 767.0kg for ages 12 through 24 months, respectively. The mean estimates of mature weight (A) to 22 and 24 months of age were not different from each other. However, they were different from the estimates based on the data to other ages. Mean estimates of rate of maturing (k) were 3.362, 3.595, 3.536, 3.421, 3.403, 3.409 and 3.411 for ages 12 through 24 months, respectively. The mean estimates of maturing rate (k) for ages 18 through 24 months of age were not significantly different from each other. However, they were different from the estimates based on the data to other ages. Correlations among estimates of A at various ages showed the highest value of 0.93 between 22 and 24 months. Correlations among estimates of k at various ages were highest ranging from 0.91 to 0.99 among 18 to 24 months. The correlations between A and k were positive and tended to decrease with the increase of the age from 0.84 for the age of 12 months to 0.10 for the age of 24 months. Thus, the estimates of growth curve parameters, A and k, suitable for genetic studies can be derived from accumulated Hanwoo bulls after 22 months of age.

Effects of Light Intensity and Nutrient Solution Strength during Short Day Treatment on the Growth and Nutrient Absorption of Kalanchoe blossfeldiana 'Rako' in Ebb and Flow System and the Accumulation of Nutrients in Growing Medium (단일처리시기의 광도와 양액농도가 Ebb and Flow 재배시스템에서 재배한 칼랑코에(Kalanchoe blossfeldiana 'Rako')의 생육, 양분흡수 및 배지 양분집적에 미치는 영향)

  • Noh, Eun-Hee;Choi, Jong-Myoung;Son, Jung-Eek
    • Journal of Bio-Environment Control
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    • v.20 no.3
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    • pp.189-196
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    • 2011
  • The objective of this study was to investigate the effects of light intensity and electrical conductivity (EC) of nutrient solution during short day treatment in an ebb and flow systems on the growth and nutrient uptake of potted Kalanchoe blossfeldiana 'Rako' and the nutrient accumulation of growing medium. Nutrient concentrations in the growing medium were also analyzed to investigate the accumulation rates of macro-nutrients such as T-N, P, K, Ca, and Mg, respectively. To achieve the objectives, plants were fed with a nutrient solution with 1.2, 1.8, or $2.4dS{\cdot}m^{-1}$ under three daily photosynthetic photon flux (PPF) of 4.26, 5.51, or $9.75mol{\cdot}m^{-2}{\cdot}d^{-1}$. Both light intensity and EC of nutrient solution significantly influenced the crop growth. The elevation of PPFs resulted in the increase of plant growth. For each light condition, plant growth, such as dry and fresh weight and leaf area, was the highest when the electrical conductivity of nutrient solution was controlled to $2.4dS{\cdot}m^{-1}$. However, growth was acceptable in the EC ranges from 1.8 to $2.4dS{\cdot}m^{-1}$. Both light intensity and EC of nutrient solution significantly influenced the uptake of nutrients in the solution tanks and the accumulation of nutrients in the growing medium. As the EC of nutrient solution was elevated, the absorption rates of $NO_3^-$, $PO_4^{-3}$, $K^+$, and $Mg^{2+}$ by crops and accumulation of those in growing medium increased, but the light intensity did not significantly influence the absorption rates. Based on the above results, the regression models were suggested for anticipating the macro-nutrient accumulations in growing medium.