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Cytotoxicity of Paraquat or Bentazone and Compensatory Effects of 3-Methylcholanthrene on the Rat Liver (Paraquat 및 Bentazone의 세포독성과 흰쥐 간에서 3-Methylcholanchrene의 독성경감효과)

  • Rim, Yo-Sup;Han, Du-Seok
    • Korean Journal of Environmental Agriculture
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    • v.20 no.3
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    • pp.155-161
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    • 2001
  • This study was carried out to investigate cytotoxicity of paraquat or bentazone on NIH 3T3 fibroblasts, toxicity of paraquat or bentazone, and compensatory effects of 3-Methylcholanthrene(3-MC) on the rat liver. In order to MTT assay, the $5.0{\times}10^4$ cell/mL of NIH 3T3 fibroblast in each well of 24 multidish were cultured. After 24 hours, the cells were treated with solution of paraquat or bentazone(1, 25, 50, 100 ${\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MTT assay were performed to evaluate the cytotoxicity of cell organelles. Paraquat or bentazone $MTT_{50}$ were 1668.97 ${\mu}M$ and 1506.97 ${\mu}M$, respectively. These $IC_{50}$ of paraquat or bentazone were decided low cytotoxicity by Borenfreund. In order to observe the toxicity and compensatory effects of paraquat or bentazone on the rat liver, Sprague-Dawley male rats were used as experimental animals and divided into paraquat or bentazone only treated group and simultaneous application group of paraquat or bentazone and 3-MC. At 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment, the animals were sacrificed by decapitation and liver were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM and Best Carmine. Under the light microscope, degenerative changes of hepatic lobules were frequently observed in portal area from 3 hrs after paraquat or bentazone treatment. All hepatic cells were induced degenerative change at 12 hrs and more severe degenerative change at 48 hrs after paraquat or bentazone treatment. Especially, hepatic cells of bentazone only treated group were distinctly showed pyknotic. Glycogen granules were increased in portal area at 3 hrs, all hepatic cells at 12 hrs and remarkably increased at 48 hrs after paraquat or bentazone treated group. But hepatic cells of bentazone only treated group were regeneration at 48 hrs from portal area and glycogen granules of hepatic cells of paraquat or bentazone and 3-MC combination treated group showed in central area only at 48 hrs. The results indicate that 3-MC may be decrease paraquat or bentazone cytotoxicity on the rat liver.

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Improvement of L-Lysine Productivity by Using Cell Fusion and Immobilized System (세포융합과 고정화 시스템을 이용한 L-Lysine의 생산성 향상)

  • Ryu, Beung-Ho;Kim, Hye-Sung;Roh, Myung-Hoon;Park, Bob-Gyu;Chung, Jong-Soon;Bai, Ki-Chul
    • Korean Journal of Food Science and Technology
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    • v.21 no.1
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    • pp.154-163
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    • 1989
  • This studies were designed to improve the productivity of L-lysine by protoplast fusion and immobilized system of fusants using strains of Brevibacterium flavum ATCC 21528, Brevibacterium lactofermentum ATCC 21086 and Corynebacterium glutamicum 820. Mutants were isolated with concentration method of $300{\mu}g/ml$ penicillin-G after treatment of $250{\mu}g/ml$ N-methyl-N-nitro-N-nitrosoguanidine. B. flavum $37-2(Hos^-,\;Kan^r,\;AEC^r)$, B. lactofermentum $6-2(Ile^-,\;Val^-,\;Str^r,\;AEC^r)$ and C. glutamicum 57-5$(Met^-,\;Thr^-,\;Rif^r,\;AEC^r)$ were isolated from mutants. Protoplasts were induced by being incubated with $500{\mu}g/ml$ lysozyme of lysis solution for 6 hr and the ratio of protoplast formation and regeneration were ranging from 97-99% and 33-37%, respectively. Fusion frequencies of fusants of BBFL 21, BCFG 37 and BCLG 59 were shown in the range from $1.25{\times}10^{-6}\;to\;5.83{\times}10^{-7}$ under the optimum conditions. The fusant BBFL 21 showed the highest productivity of $411.1\;ng/ml{\cdot}hr$ L-lysine in the lysine productivity broth at $30^{\circ}C$ for 72hr. In the immobilization systems, fusant BBFL 21 was employed in various polymer matrices such as sodium alginate, polyacrylamide, agar and ${\alpha}-carrageena$. The immobilization of sodium alginate showed the highest productivity of $413\;ng/ml{\cdot}hr$ L-lysine in the batch system. Continuous fermentation of immobilization system by using tube fermentor was produced the highest productivity $416.7\;ng/ml{\cdot}hr $ L-lysine under optimum condition.

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The Microleakage in Class V Cavity Based on Different Kinds of Several Dentin Adhesive and Composite Resin (5급 와동에서 수종의 상아질 접착제와 복합레진의 종류에 따른 미세누출)

  • Choi, Yoo-Seok;Jeong, Soon-Jeong;Jeong, Moon-Jin;Ahn, Yong-Soon;Park, Seon-Nyeo;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.330-338
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    • 2013
  • In order to explore microleakage in class V cavity based on different kinds of several dentin adhesive and composite resin, 2 kinds of composite resin was restored and exposed after applying 4 kinds of dentin adhesives. Deposited in methylene blue solution for 4 hours and cut in parallel with tooth longitudinal axis. By observing dye penetration level of enamel and dentin margins of each restored resin following conclusion was obtained. 1. In composite resin Filtek Z350XT Universal (3M/ESPE Dental Products, USA) in enamel margin, Easy Bond (3M/ESPE Dental Products) showed the lowest microleakage and this leakage was represented to be high in the order of Single Bond 2 (3M/ESPE Dental Products), Scotchbond Multi-Purpose (3M/ESPE Dental Products) and Cearfil SE Bond (Kuraray Medical Inc., Japan). In case of Filtek Z350XT Flowable (3M/ESPE Dental Products), Scotchbond Multi-Purpose showed the lowest microleakage and this leakage was represented to be high in the order of Single Bond 2, Clearfil SE Bond and Easy Bond. 2. In case of Filtek Z350XT Universal in dentin margin, Easy Bond showed the lowest microleakage and this leakage was represented to be high in the order of Scotchbond Multi-Purpose, Single Bond 2 and Clearfil SE Bond. In case of Filtek Z350XT Flowable, Scotchbond Multi-Purpose and Single Bond showed the lowest microleakage and this leakage was represented to be high in the order of Clearfil SE Bond and Easy Bond. 3. In all the groups excepting S-U group (Single Bond 2+Filtek Z350XT Universal), enamel margin showed more higher microleakage than that of dentin margin. 4. There was a difference between enamel and dentin margin among each group but it was not significant statistically (p>0.05). When summarizing this result, it is considered that composite resin and dentin adhesive could be applied selectively and particularly in case of applying 1-step self-etching dentin adhesive, this method would be advantageous for manipulation convenience and shortening of operation time.

Spectrophotometric Determination of Aluminium Ion in Drinking Water by Flow Injection Analysis (흐름주입분석법에 의한 음용수 중 알루미늄 이온의 분광광도법 정량)

  • Choi, Yong-Wook;Jin, Jae-Young
    • Journal of the Korean Chemical Society
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    • v.44 no.5
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    • pp.422-428
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    • 2000
  • Optimum analytical conditions of the aluminium ion were established by flow injection analysis. Eriochrome Cyanine R(ECR) dye reacts with the aluminium ion at pH 6.0 to form a complex that exhibits maximum absorption at 535 nm. Reaction conditions including the mixing and the reaction coil length, the concentration and the pH of the buffer solutio, temperature, and injection loop volume were optimized to intro-duce this reaction into flow injection analysis. The results were as follows. A mixing coil length of 0.5 m and a reaction coil length of 4.0 m, the pH 6.0 and 1M of acetate buffer solution, the ECR concentration of 0.56 mM, the reaction temperature of 40$^{\circ}C$, the injection loop volume of 300${\mu}L$ were chosen as optimum conditions. Under these conditions the detection limit of the aluminiumion was less than 0.05 mg/L and the repeatability was better than 1%. A sampling frequency of 24 times for an hour was achieved. Interfering ions such as $F^-$, HP$O_4^{2-}$, $Fe^{2+}$, $Fe^{3+}$, $Mn^{2+}$, and other anions were tested, interference did not occur up to 1,000mg/L of ion concentration and up to 2,CO0mg/L of sulfate ion con-centration. This method was applied for the determination of aluminium ion in tap water and ground water of Jeonju and the Gochang area. The results showed that the aluminium residual in tap water of the Jeonju area was at a mean of 0.478mg/L and that in tap water of the Gochang area was at a mean of 0.278mg/L. Aluminium ion residual of the tap waters in the Jeonju area was higher level than that in the Gochang area. Aluminium residual in the ground water of the Jeonju area was 0.386 mg/L and was lower compared to 0.564 mg/L for the Gochang area.

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The Effects of Medicinal Herbs Extracts on Estrogen-like Activities and Osteoblast Proliferation and Differentiation (한약재 추출물의 에스트로겐 유사활성 및 조골세포 증식과 분화에 미치는 영향)

  • Kim, Mihyang;Kim, Bokyung;Kim, Jae-Deog;Kang, A-Ram;Lee, Chang-Eun;Seo, Jungmin;Lee, Dong-Geun;Jo, Jung-Kwon;Kim, Yuck Yong;Yu, Ki Hwan;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.27 no.4
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    • pp.456-463
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    • 2017
  • The purpose of this study was to investigate the effect of 3 types of medicinal herbs (Glycyrrhizae radix, Astragali radix and Dioscorea rhizoma) extracts on estrogen-like activities, proliferation and differentiation in osteoblast. Human breast cancer cell line MCF7 was transfected using an estrogen responsive luciferase reporter plasmid for measure the estrogen-like activity. Estrogen-like activities of extracts were in the range of 1.11~5.73 fold to that of negative control. The extract of G. radix showed the strongest estrogen-like activities. The estrogen-like activities of 50 and $500{\mu}g/ml$ extracts of G. radix were similar to that of $10^{-8}$ and $10^{-7}$ M standard solution ($17{\beta}-estradiol$), respectively. G. radix extract showed no cytotoxicity against osteoblast MC3T3-E1 cells at $1{\sim}1,000{\mu}g/ml$. The extract of A. radix showed no significant proliferation of osteoblast. However, the extract of G. radix and D. rhizome showed maximum 148% and 133% proliferation effects. The extract of G. radix also increased alkaline phosphatase activity and the maximum was 122% at $100{\mu}g/ml$ compared to that of control. The nodule formation by the method of the Alizarin red S staining increased compared to control. These results suggest that G. radix is able to perform the bone formation and prevent osteoporosis.

The Study on the Methylmercury Analysis and the Monitoring of Total Mercury and Methylmercury in Fish (어류 중 메틸수은 분석법 확립 및 모니터링)

  • Kim, Hee-Yun;Chung, So-Young;Sho, You-Sub;Oh, Geum-Soon;Park, Seong-Soo;Suh, Jung-Hyuk;Lee, Eun-Ju;Lee, Yoon-Dong;Choi, Woo-Jeong;Eom, Ji-Yoon;Song, Min-Soo;Lee, Jong-Ok;Woo, Gun-Jo
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.882-888
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    • 2005
  • Procedure for analysis of methylmercury in fish was developed, involving addition of HCl, extraction with toluene, and clean-up using L-cystein solution. Obtained extract is analyzed by gas chromatography with electron capture detector using Ulbon HR-Thermon-Hg column. Detection limit and recovery of the method were 0.005mg/kg (expressed as Hg), 98-107 (103%), respectively. Total mercury and methylmercury concentrations in 175 commercial fish samples ranged from [mean-max (mean), unit: mg/kg]: 0.014-1.200 (0.270) and 0.006-0.901 (0.168) in tuna-fish, 0.020-0.934 (0.323) and 0.012-0.553 (0.149) in martin-fish, 0.082-0.782 (0.391) and 0.040-0.436(0.201) in shark, 0,023-0.031 (0.026) and 0,013-0.018 (0.015) in salmon, 0.098-0.193 (0.133) and 0.031-0.015(0.090) in tilefish, and 0,031-0.214 (0.089) and 0.016-0.093 (0.042) in canned tuna respectively. No sample of analyzed fish exceeded 1.0mg/kg wet wt., limit for methylmercury established by Codex. In all species examined, estimated weekly intake was lower than Provisional Tolerable Weekly Intake recommended by the JECFA (the Joint FAO/WHO Expert Committee on Food Additives).

Seed Germination, Seedling Grwoth and Optimal Seedling Density of Phellodendron amurense Rupr. in Nursery (황벽나무의 종자발아(種子發芽)와 묘목생장(苗木生長) 및 적정생육밀도(適正生育密度)에 관(關)한 연구(硏究))

  • Goo, Gwan-Hyo;Lee, Kang-Young;Youn, Ki-Sik;Lee, Chong-Kyu
    • Journal of Korean Society of Forest Science
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    • v.86 no.4
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    • pp.443-449
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    • 1997
  • We examined the effect of pretreatments on seed germination, seedling growth, optimal seedling density of Phellodendron amurense. The seeds were carefully purified after collection in late Oct. from natural population at Mt. Chiri. For the evaluation of pretreatment effects, seeds were sowed in an experimental nursery after being stored in open ground($2{\sim}8^{\circ}C$) with soaking in 20% $H_2O_2$, 50-fold pon-pon solution, cold moist stratification in refrigerator($2{\sim}4^{\circ}C$) and dry storage in roam temperature($4{\sim}18^{\circ}C$), respectively, For the effect of growth density of seedlings, seedlings germinated in nursery were precisely controlled with 53, 104, 220, 380 seedlings per $m^2$. Among pretreatments for the promotion of germination rate, pon-pon treatment(68.8%) was highest, the others were higher than that in dry storage(2.3%), and these seed pretreatment methods were significantly different at 1% level. The height growth of seedlings showed the most vigor from Jun.12 to Jul. 8, and from Jul.20 to Aug. 2, and it reached to 72.8% of height growth. In case of 1-0 seedling, although seedling density per $m^2$ was increased, seedling growth was decreased. The optimal seedling density was 104 seedlings per $m^2$. The standard seedlings for plantation were above height 55cm, root collar diameter 8.1mm, and root length 21cm. These results demonstrated that seed of P. amurense requires stratification for the germination promotion.

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Cryopreservation of Bovine IVM/IVF/IVC Blastocysts by Vitrification (체외성숙, 체외수정 및 체외배양에서 생산된 소 배반포기배의 초자화 동결)

  • Nam, H.K.;Kim, E.Y.;Lee, K.S.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.231-238
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    • 1999
  • The objective of this study was to examine the effect of developmental stage and embryo age of in vitro produced bovine blastocysts after vitrification and thawing. In vitro cultured day 8 blastocysts after IVF were equilibrated 20% ethylene glycol (EG) for 3 min. and were vitrified using EFS40, which is consisted of 40% EG, 18% ficoll, 0.3M sucrose and 10% FBS added in mDPBS for 30 sec. before being plunged into $LN_2$. Also, survival in vitro was assessed by re-expansion and hatching or hatched at 24 hand 48 h postwarming, respectively. The results obtained in these experiments were summarized as follows; 1) When the embryos were cultured for 8 day after IVF, 41.0% of the cleaved embryos developed to the blastocysts (early; 7.6%, expanded; 22.9%, hatching; 4.6% and hatched; 5.9%). 2) When the embryos were exposed or vitrified to the freezing solution, the re-expansion of vitrified embryos (73.3%) was significantly lower than that of control and exposed embryos (100, 97.0%) (p<0.05). But the formation rate of hatching or hatched blastocysts of vitrified embryos (66.7, 46.7%) at 48h after thawing was similar to that of exposed embryos (66.7, 39.4%) but not control (100, 100%) (p<0.01). However, in the total cell numbers of those developed hatched blastocysts, there were not significantly different among the treatment groups. 3) When the embryo survival rates by different developmental stage were examined, the re-expansion was not different among the groups $(64.5{\sim}75.6%)$. After warming 48 h, the hatching and hatched formation of early blastocysts (25.8, 9.7%) was significantly lower than those of expanded (69.7, 39.4%) and hatching blastocysts (53.3, 43.3%) (p<0.05). 4) In addition, when the expanded blastocysts at day 7, 8 and 9 were vitrified, the re-expansion of day 8 and 9 embryos was significantly lower than that of day 7 (day 7; 93.9%, day 8; 75.8% and day 9; 87.5%) (p<0.05). However, the rates of development to hatched blastocysts were no difference among the groups (day 7; 36.4%, day 8; 36.4% and day 9; 31.3%). These results suggested that in vitro produced expanded or hatching blastocysts can be efficiently cryopreserved by the two-step vitrification method using EFS40.

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A comparative study on the fracture behavior of zironia, glass infiltrated alumina and PFM full crown system (지르코니아, 유리침투알루미나 및 PFM 전부관 시스템의 파절 경향에 관한 비교연구)

  • Lee, Sang-Hyeok;Ahn, Jin-Soo;Kim, Myung-Ho;Lim, Bum-Soon
    • The Journal of Korean Academy of Prosthodontics
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    • v.50 no.4
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    • pp.235-242
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    • 2012
  • Purpose: The purpose of this study was to compare the fracture behavior of Zironia, glass infiltrated Alumina and PFM full crown system. Materials and methods: Fifteen crowns for each of 3 experimental groups (Zironia, glass infiltrated Alumina and PFM full crown) were made by the conventional method. The crowns mounted on the testing jig were inclined in 30 degrees to the long axis of the tooth and the universal testing machine was used to measure the fracture strength. Results: 1. The mean fracture strengths were $588.3{\pm}49.6MPa$ for zirconia system, $569.1{\pm}61.8MPa$ for PFM system and $551.0{\pm}76.5MPa$ for glass-infiltrated alumina system (P>.05). 2. The mean shear bond strengths were $25.5{\pm}5.6MPa$ for zirconia system, $38.9{\pm}5.0MPa$ for Ni-Cr alloy system and $39.4{\pm}5.1MPa$ for glass-infiltrated alumina system. 3. The chemical bonding was observed at interfaces between PFM or glass-infiltrated alumina and veneering porcelain, however, no chemical bonding was observed at interface between zirconia and veneering porcelain. Conclusion: With the study, the fracture strengths of PFM crown system had a higher fracture strength than conventional zirconia system crown and glass-infiltrated alumina crowns. and than the shear bond strengths glass-infiltrated alumina system had a higher shear bond strength than conventional PFM system and zirconia system.

A Study of EFFECT and MECHANISM of IL-2 on SURVIVAL of EOSINOPHILS (Interleukin-2가 호산구 생존에 미치는 영향과 가전에 관한 연구)

  • Kim, Hyo-Seok;Lee, Young-Mok;Choi, Young-Soo;Kim, Kyung-Ho;Im, Geon-Il;Moon, Seung-Hyug;Jeong, Sung-Whan;Kim, Hyeon-Tae;Uh, Soo-Taek;Kim, Yong-Hun;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.3
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    • pp.348-358
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    • 1996
  • Background : Interleukin-5 (IL-5) is responsible for eosinophilia in allergic diseases. In allergic bronchial asthma, there is a correlation between the extent of eosinophil infiltration in bronchial mucosa and IL-5 concentrations. In addition, IL-2 concentration is elevated in the airways and associated with eosinophilia in symptomatic patients with bronchial asthma. In animal studies, IL-2 can induce eosinophilia by increasing the synthesis of IL-5, however, it is still unknown how IL-2 can induce eosinophila in human being. The aim of this study is to evaluation the effect and mechanism of IL-2 on prolongation of eosinophil survival. Methods : After purifiing the eosinophils from the venous blood of allergic patients with eosinophilia, we measured the survival rates of eosinophils using trypan blue dye exclusion test, and the number of eosinophils with Randolp's solution. We compared the survival rates of eosinophils in the presence of IL-2 or IL-5. Neutralizing antibody for IL-5 was added in IL-2 treated eosinophils to reveal whether IL-2 induced prolongation of eosinophil survival was mediated by IL-5. We checked IL-5 m-RNA expression of lymphocytes in the presence of IL-2 by using Reverse transcription-Polymerase chain reaction (RT-PCR) method to revealed the effect of IL-2 on IL-5 m-RNA expression on lymphocyte. $\alpha$ and $\beta$ IL-2 receptors were measured on eosinophils and lymphocytes with flow-cytometer after stimulated with IL-2. Results : 1) Eosinophil survival rates increased dose dependently on IL-5 and IL-2. 2) The eosinophil survival rates increased by IL-2 were not inhibited by the pretreatment with neutralizing antibody for IL-5. 3) IL-5 m-RNA was not expressed on lymphocytes by the treatment with IL-2 up to 96 hours. 4) IL-2 upregulate the expression of IL-$2R{\alpha}$ on eosinophils, instead of no effect on the expression of IL-$2R{\beta}$. Conclusion: Interleukin-2 had the enhancing effect on the survival rates of eosinophils. The mechanism behind IL-2 induced eosinophilia might be the increment of IL-2 receptors on eosinophils rather than IL-5 synthesis by lymphocytes.

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