动物营养学报 >

2021 , Vol. 33 >Issue 11: 6534 - 6547

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.11.052

研究简报 SHORT COMMUNICATION

饲料花生四烯酸含量对子二代中华鲟成活与生长、抗氧化能力、组织花生四烯酸代谢酶活性及肠道菌群组成的影响

  • 吴金平 ,
  • 杨代勤 ,
  • 杜浩 ,
  • 罗江 ,
  • 熊伟 ,
  • 刘源 ,
  • 危起伟 ,
  • 朱建强
展开
  • 1. 长江大学农学院, 荆州 434025;
    2. 中国水产科学研究院长江水产研究所, 农业农村部淡水生物多样性保护重点实验室, 武汉 430223
吴金平(1989-),男,湖北武汉人,助理研究员,博士研究生,从事鲟鱼营养与饲料研究。E-mail:Wujinping@yfi.ac.cn

收稿日期: 2021-03-30

  网络出版日期: 2021-11-10

基金资助

中国水产科学研究院基本科研业务费项目"国家农业科学渔业资源环境武汉观测实验站科研监测与评价"（2020JBF01）；湿地生态与农业利用教育部工程研究中心开放基金（KFT201904）

收起

Effects of Dietary Arachidonic Acid Content on Survival and Growth, Antioxidant Capacity, Arachidonic Acid Metabolic Enzyme Activities and Intestinal Microbial Composition of F2 Generation Chinese Sturgeon

  • WU Jinping ,
  • YANG Daiqin ,
  • DU Hao ,
  • LUO Jiang ,
  • XIONG Wei ,
  • LIU Yuan ,
  • WEI Qiwei ,
  • ZHU Jianqiang
Expand
  • 1. College of Agriculture, Yangtze University, Jinzhou 434025, China;
    2. Key Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture and Rural Affairs of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China

Received date: 2021-03-30

  Online published: 2021-11-10

Supported by

 

Fold

摘要

本试验旨在探究饲料花生四烯酸（ARA）含量对子二代中华鲟成活与生长、抗氧化能力、组织ARA代谢酶活性及肠道菌群组成的影响。通过在基础饲料中添加0、0.5%、1.0%和2.0%的ARA纯化油（ARA占总脂肪酸的44.88%），制成4种等氮等能的试验饲料，饲料ARA含量分别为0.18%、0.53%、0.96%和1.89%。用4种试验饲料投喂初始体质量为（4.17±0.22） kg的子二代中华鲟22周，每种饲料投喂3个重复，每个重复18尾鱼。结果表明：试验鱼的增重率与成活率不受饲料ARA含量的显著影响（P>0.05）。肝脏超氧化物歧化酶（SOD）、过氧化氢酶（CAT）与谷胱甘肽硫转移酶（GST）活性随饲料ARA含量的升高而升高，丙二醛（MDA）含量随饲料ARA含量的升高而下降。血清SOD、CAT、GST活性与MDA含量不受饲料ARA含量的显著影响（P>0.05）。性腺中，1.89% ARA组环氧合酶-2（COX-2）与细胞色素氧化酶P450（CYP450）活性显著高于0.18%与0.53% ARA组（P<0.05），与0.96% ARA组无显著差异（P>0.05）；0.96% ARA组环氧合酶-1（COX-1）活性显著高于0.18%与0.53% ARA组（P<0.05）；1.96% ARA组脂氧合酶（LOX）活性显著高于0.18% ARA组（P<0.05）。肝脏中，COX-1、COX-2与LOX活性均以1.89% ARA组最高，与0.96% ARA组无显著差异（P>0.05）；0.18% ARA组CYP450活性显著低于0.96%与1.89% ARA组（P<0.05）。0.18% ARA组Chao1指数、ACE指数、Shannon指数与Observed-species数量显著高于0.53% ARA组（P<0.05），与0.96%及1.89% ARA组无显著差异（P>0.05）。Simpson指数与PD-whole tree指数不受饲料ARA含量的显著影响（P>0.05）。上述结果表明，饲料中添加1.0%的ARA纯化油使饲料ARA含量为0.96%时对子二代中华鲟的生长与存活无显著影响，但能增强鱼体的抗氧化能力、组织中ARA代谢酶活性及调控肠道菌群多样性。

关键词： 花生四烯酸; 中华鲟; 生长; 存活; 抗氧化能力; 花生四烯酸代谢酶; 菌群多样性

本文引用格式

吴金平 , 杨代勤 , 杜浩 , 罗江 , 熊伟 , 刘源 , 危起伟 , 朱建强 . 饲料花生四烯酸含量对子二代中华鲟成活与生长、抗氧化能力、组织花生四烯酸代谢酶活性及肠道菌群组成的影响[J]. 动物营养学报, 2021 , 33(11) : 6534 -6547 . DOI: 10.3969/j.issn.1006-267x.2021.11.052

Abstract

The effects of arachidonic acid (ARA) on survival and growth, antioxidant capacity, tissue ARA metabolic enzyme activities and intestinal microbial composition of the F2 generation Chinese sturgeon were evaluated in this experiment. Four isonitrogenous and isoenergetic experimental diets were formulated by adding 0, 0.5%, 1.0% and 2.0% purified ARA oil (43% ARA in total fatty acids) in the basal diet, and dietary arachidonic acid content was 0.18%, 0.53%, 0.96% and 1.89%, respectively. The F2 generation Chinese sturgeon with an initial body weight of (4.17±0.22) kg were fed for 22 weeks with 3 replicates per diet treatment, and 18 fish per replicate. The results showed that the weight gain rate and survival rate were not significantly affected by dietary ARA content (P>0.05). The activities of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST) in the liver were increased with dietary ARA content increasing, while the content of malonic aldehyde (MDA) in the liver was decreased with dietary ARA content increasing. The activities of SOD, CAT, GST and the content of MDA in the serum were not significantly affected by dietary ARA content (P>0.05). In the gonad, the activities of cyclooxygenase-2 (COX-2) and cytochrome P450 (CYP450) of the 1.89% ARA group were significantly higher than those of the 0.18% and 0.53% ARA groups (P<0.05), but there were no significant differences between the 1.89% and 0.96% ARA groups (P>0.05); the cyclooxygenase-1 (COX-1) activity of the 0.96% ARA group was significantly higher than that of the 0.18% and 0.53% ARA groups (P<0.05); the lipoxygenase (LOX) activity of the 1.89% ARA group was significantly higher than that of the 0.18% ARA group (P<0.05). In the liver, the activities of COX-1, COX-2 and LOX were the highest in the 1.89% ARA group, and there were no significant differences between the 0.96% and 1.89% ARA groups (P>0.05); the CYP450 activity of the 0.18% ARA group was significantly lower than that of the 0.96% and 1.89% groups (P<0.05). The Chao1 index, ACE index, Shannon index and Observed-species number of the 0.18% ARA group were significantly higher than those of the 0.53% ARA group (P<0.05), and there were no significant differences among the 0.18%, 0.96% and 1.89% ARA groups (P>0.05). The Simpson index and PD-whole tree index were not significantly affected by dietary ARA content (P>0.05). These results indicates that dietary with 1.0% ARA has no significant effects on the growth and survival of F2 Chinese sturgeon, but can enhance the antioxidant capacity, tissue ARA metabolic enzyme activities and regulate the diversity of intestinal microbes.

Key words： arachidonic acid; Chinese sturgeon; growth; survival; antioxidant capacity; ARA metabolic enzymes; microbial diversity

参考文献

[1] HALVER J E.Fish nutrition[M].2nd ed.San Diego:Academic Press, 1989:153-218.
[2] SARGENT J R, BELL J G, BELL M V, et al.Requirement criteria for essential fatty acids[J].Journal of Applied Ichthyology, 1995, 11(3/4):183-198.
[3] HALVER J E, HARDY R W.Fish nutrition[M].3rd ed.Pittsburgh:Academic Press, 2002:181-257.
[4] BELL J G, SARGENT J R.Arachidonic acid in aquaculture feeds:current status and future opportunities[J].Aquaculture, 2003, 218(1/2/3/4):491-499.
[5] 刘镜恪, 陈晓林, 李岿然, 等.实验微粒饲料中花生四烯酸含量对牙鲆(Paralichthys olivaceus)仔稚鱼生长、存活的影响[J].海洋与湖沼, 2005, 36(5):418-422. LIU J K, CHEN X L, LI K R, et al.Effects of different contents of arachidonic acid in experimental microdiets on growth and survival of larval Japanese flounder Paralichthys olivaceus[J].Oceanologia et Limnologia Sinica, 2005, 36(5):418-422.(in Chinese)
[6] BAE J Y, KIM D J, YOO K Y, et al.Effects of dietary arachidonic acid (20:4n-6) levels on growth performance and fatty acid composition of juvenile eel, Anguilla japonica[J].Asian-Australasian Journal of Animal Sciences, 2010, 23(4):508-514.  
[7] XU H G, AI Q H, MAI K S, et al.Effects of dietary arachidonic acid on growth performance, survival, immune response and tissue fatty acid composition of juvenile Japanese seabass, Lateolabrax japonicus[J].Aquacultue, 2010, 307(1/2):75-82.
[8] 谢奉军.大黄鱼仔稚鱼氨基酸及脂肪酸营养生理的研究[D].博士学位论文.青岛:中国海洋大学, 2011. XIE F J.Studies on nutritional physiology of amino acid and fattr acid for large yellow croaker (Pseudosciaena crocea) larve[D].Ph.D.Thesis.Qingdao:Ocean University of China, 2011.(in Chinese)
[9] LUO Z, TAN X Y, LI X D, et al.Effect of dietary arachidonic acid levels on growth performance, hepatic fatty acid profile, intermediary metabolism and antioxidant responses for juvenile Synechogobius hasta[J].Aquacultue Nuntrition, 2012, 18(3):340-348.  
[10] 王成强, 梁萌青, 徐后国, 等.大规格鲈鱼(Lateolabrax japonicas)对饲料中花生四烯酸的需求量[J].渔业科学进展, 2016, 37(5):46-55. WANG C Q, LIANG M Q, XU H G, et al.Requirement of arachidonic acid in adult Japanese seabass (Lateolabrax japonicas)[J].Progress in Fishery Science, 2016, 37(5):46-55.(in Chinese)
[11] KOVEN W, VAN ANHOLT R, LUTZKY S, et al.The effect of dietary arachidonic acid on growth, survival, and cortisol levels in different-age gilthead seabream larvae (Sparus auratus) exposed to handling or daily salinity change[J].Aquaculture, 2003, 228(1/2/3/4):307-320.
[12] BRANSDEN M P, COBCROFT J M, BATTAGLENE S C, et al.Dietary arachidonic acid alters tissue fatty acid profile, whole body eicosanoid production and resistance to hypersaline challenge in larvae of the temperate marine fish, striped trumpeter (Latris lineata)[J].Fish Physiology and Biochemistry, 2004, 30:241-256.
[13] TORRECILLAS S, BETANCOR M B, CABALLERO M J, et al.Supplementation of arachidonic acid rich oil in European sea bass juveniles (Dicentrarchus labrax) diets:effects on growth performance, tissue fatty acid profile and lipid metabolism[J].Fish Physiology and Biochemistry, 2018, 44(1):283-300.  
[14] ARAÚJO B C, FLORES-GALVEZ K, HONJI R M, et al.Arachidonic acid effects on the overall performance, fatty acid profile, hepatopancreas morphology and lipid-relevant genes in Litopenaeus vannamei juveniles[J].Aquaculture, 2020, 523:735207.
[15] NORAMBUENA F, ESTÉVEZ A, MAÑANÓS E, et al.Effects of graded levels of arachidonic acid on the reproductive physiology of Senegalese sole (Solea senegalensis):fatty acid composition, prostaglandins and steroid levels in the blood of broodstock bred in captivity[J].General and Comparative Endocrinology, 2013, 191:92-101.
[16] XU H G, CAO L, ZHANG Y Q, et al.Dietary arachidonic acid differentially regulates the gonadal steroidogenesis in the marine teleost, tongue sole (Cynoglossus semilaevis), depending on fish gender and maturation stage[J].Aquaculture, 2017, 468(Part 1):378-385.
[17] 张圆琴, 徐后国, 曹林, 等.饲料中花生四烯酸对发育前期大菱鲆亲鱼性类固醇激素合成的影响[J].水产学报, 2017, 41(4):588-601. ZHANG Y Q, XU H G, CAO L, et al.Effects of dietary arachidonic acid on the sex steroid hormone synthesis in turbot broodstock before maturation[J].Journal of Fisheries of China, 2017, 41(4):588-601.(in Chinese)
[18] COPEMAN L A, PARRISH C C, BROWN J A, et al.Effects of docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limanda ferruginea):a live food enrichment experiment[J].Aquacultue, 2002, 210(1/2/3/4):285-304.
[19] VILLALTA M, ESTÉVEZ A, BRANSDEN M P, et al.Arachidonic acid, arachidonic/eicosapentaenoic acid ratio, stearidonic acid and eicosanoids are involved in dietary-induced albinism in Senegal sole (Solea senegalensis)[J].Aquacultue Nuntrition, 2008, 14(2):120-128.  
[20] LUND I, STEENFELDT S J, HANSEN B W.Influence of dietary arachidonic acid combined with light intensity and tank colour on pigmentation of common sole (Solea solea L.) larvae[J].Aquacultue, 2010, 308(3/4):154-159.
[21] LI Q F, AI Q H, MAI K S, et al.In vitro effects of arachidonic acid on immune functions of head kidney macrophages isolated from large yellow croaker (Larmichthys crocea)[J].Aquacultue, 2012, 330/333:47-53.
[22] ESTÉVEZ A, MCEVORY L A, BELL J G, et al.Growth, survival, lipid composition and pigmentation of turbot (Scophthalmus maximus) larvae fed live-prey enriched in arachidonic and eicosapentaenoic acids[J].Aquacultue, 1999, 180(3/4):321-343.
[23] ASIL S M, KENARI A A, MIYANJI G R, et al.The influence of dietary arachidonic acid on growth, reproductive performance, and fatty acid composition of ovary, egg and larvae in an anabantid model fish, blue gourami (Trichopodus trichopterus;Pallas, 1770)[J].Aquacultue, 2017, 476:8-18.
[24] NHAN H T, NHU T Q, DUC P M, et al.Effects of dietary arachidonic acid on final maturation, spawning and composition of gonad of black sea urchin Diadema setosum (Leske, 1778)[J].Aquaculture Nutrition, 2020, 26(5):1771-1779.  
[25] TIAN J J, LEI C X, JI H, et al.Role of cyclooxygenase-mediated metabolites in lipid metabolism and expression of some immune-related genes in juvenile grass carp (Ctenopharyngodon idellus) fed arachidonic acid[J].Fish Physiology and Biochemistry, 2017, 43(3):703-717.  
[26] FEI S Z, CHEN Z, XIA Y.Effects of dietary arachidonic acid on reproduction performance, tissue fatty acid profile and gonadal steroidogenesis in female yellow catfish Pelteobagrus fulvidraco[J].Aquaculture Nutrition, 2020, 27(3):700-711.
[27] 马红娜.饲料脂肪和花生四烯酸水平对黄颡鱼生长性能、脂肪酸组成和脂肪代谢的影响[D].硕士学位论文.宁波:宁波大学, 2018. MA H N.Effects of dietary lipid and arachidonic acid levels on growth performance, fatty acid profiles and lipid metabolism of juvenile yellow catfish (Pelteobagrus fulvidraco)[D].Master's Thesis.Ningbo:Ningbo University, 2018.(in Chinese)
[28] WIRTH M, KIRSCHBAUM F, GESSNER J, et al.Fatty acid composition in sturgeon caviar from different species:comparing wild and farmed origins[J].International Review Hydrobiology, 2002, 87(5/6):629-636.
[29] WANG J H, WEI Q W, ZOU Y C.Conservation strategies for the Chinese sturgeon, Acipenser sinensis:an overview on 30 years of practices and future needs[J].Journal of Applied Ichthyology, 2011, 27(2):176-180.  
[30] QIAO Y, TANG X, BROSSE S, et al.Chinese sturgeon (Acipenser sinensis) in the Yangtze river:a hydroacoustic assessment of fish location and abundance on the last spawning ground[J].Journal of Applied Ichthyology, 2006, 22(Suppl.1):140-144.
[31] 危起伟, 李罗新, 杜浩, 等.中华鲟全人工繁殖技术研究[J].中国水产科学, 2013, 20(1):1-11. WEI Q W, LI L X, DU H, et al.Research on technology for controlled propagation of cultured Chinese sturgeon (Acipenser sinensis)[J].Journal of Fishery Sciences of China, 2013, 20(1):1-11.(in Chinese)
[32] ZHOU H, LENG X Q, TAN Q S, et al.Identification of key nutrients for gonadal development by comparative analysis of proximate composition and fatty/amino acid profile in tissues and eggs of Chinese sturgeon (Acipenser sinensis Gray, 1835)[J].Journal of Applied Ichthyology, 2017, 33(5):885-891.  
[33] LIU J J, LI Q F, GUO B F, et al.Selective utilization of fatty acids by Chinese sturgeon (Acipenser sinensis Gray, 1835) during gonadal development and first year juvenile growth[J].Journal of Applied Ichthyology, 2017, 30(6):1181-1185.
[34] LENG X Q, ZHOU H, TAN Q S, et al.Integrated metabolomic and transcriptomic analyses suggest that high dietary lipid levels facilitate ovary development through the enhanced arachidonic acid metabolism, cholesterol biosynthesis and steroid hormone synthesis in Chinese sturgeon (Acipenser sinensis)[J].The British Journal of Nutrition, 2019, 122(11):1230-1241.  
[35] MAGOČ T, SALZBERG S L.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics, 2011, 27(27):2957-2963.
[36] BOKULICH N A, SUBRAMANIAN S, FAITH J J, et al.Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing[J].Nature Methods, 2013, 10(1):57-59.  
[37] CAPORASO J G, KUCZYNSKI J, STOMBAUGH J, et al.QIIME allows analysis of high-throughput community sequencing data[J].Nature Methods, 2010:335-336.
[38] ROGNES T, FLOURI T, NICHOLS B, et al.VSEARCH:a versatile open source tool for metagenomics[J].PeerJ, 2016, 4:e2584.
[39] EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods, 2013, 10(10):996-998.  
[40] WANG Q, GARRITY G M, TIEDJE J M, et al.Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J].Applied and Environmental Microbiology, 2007, 73(16):5261-5267.  
[41] 王成强, 王际英, 黄炳山, 等.饲料花生四烯酸水平对珍珠龙胆石斑鱼幼鱼生长性能、抗氧化能力、血清生化指标以及肝脏和肌肉脂肪酸组成的影响[J].动物营养学报, 2018, 30(9):3567-3580. WANG C Q, WANG J Y, HUANG B S, et al.Effects of dietary arachidonic acid level on growth performance, antioxidant ability, serum biochemical parameters and fatty acid composition in liver and muscle of juvenile hybrid grouper (Epinephelus fuscoguttatus♀×Epinephelus lanceolatus)[J].Chinese Journal of Animal Nutrition, 2018, 30(9):3567-3580.(in Chinese)
[42] 李广, 李世顺, 李敏, 等.饲料中花生四烯酸对中间球海胆存活、生长和肠道菌群组成的影响[J].中国渔业质量与标准, 2018, 8(3):42-51. LI G, LI S X, LI M, et al.Effects of dietary arachidonic acid on survival and growth performance and the intestinal bacterial profile of adult sea urchin (Strongylocentrotus intermedius)[J].Chinese Fishery Quality and Standards, 2018, 8(3):42-51.(in Chinese)
[43] CHEE W L, TURCHINI G M, TEOH C Y, et al.Dietary arachidonic acid and the impact on growth performance, health and tissues fatty acids in Malabar red snapper (Lutjanus malabaricus) fingerlings[J].Aquaculture, 2020, 519:734757.
[44] BRANSDEN M P, BUTTERFIELD G M, WALDEN J, et al.Tank colour and dietary arachidonic acid affects pigmentation, eicosanoid production and tissue fatty acid profile of larval Atlantic cod (Gadus morhua)[J].Aquaculture, 2005, 250(1/2):328-340.
[45] 危起伟.中华鲟保护生物学[M].北京:科学出版社, 2019:53-54. WEI Q W.Conservation biology of Chinese sturgeon (Acipenser sinensis)[M].Beijing:Science Press, 2019:53-54.(in Chinese)
[46] SHI X C, JIN A, SUN J, et al.α-lipoic acid ameliorates n-3 highly-unsaturated fatty acids induced lipid peroxidation via regulating antioxidant defenses in grass carp (Ctenopharyngodon idellus)[J].Fish & Shellfish Immunology, 2017, 67:359-367.
[47] CHANDRAN R, SIVAKUMAR A A, MOHANDASS S, et al.Effect of cadmium and zinc on antioxidant enzyme activity in the gastropod, Achatina fulica[J].Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology, 2005, 140(3/4):422-426.
[48] 王成强, 王际英, 李宝山, 等.饲料中花生四烯酸含量对刺参生长性能、抗氧化能力及脂肪酸代谢的影响[J].中国水产科学, 2018, 25(3):555-566. WANG C Q, WANG J Y, LI B S, et al.Effects of dietary arachidonic acid on the growth performance, anti-oxidant capacity, and fatty acid metabolism of sea cucumber (Apostichopus japonicus)[J].Journal of Fishery Sciences of China, 2018, 25(3):555-566.(in Chinese)
[49] FUNK C D.Prostaglandins and leukotrienes:advances in eicosanoid biology[J].Science, 2001, 294(5548):1871-1875.  
[50] YIN H Y, ZHOU Y H, ZHU M J, et al.Role of mitochondria in programmed cell death mediated by arachidonic acid-derived eicosanoids[J].Mitochondrion, 2013, 13(3):209-224.  
[51] 田晶晶.花生四烯酸(ARA)对草鱼脂质代谢的影响及其机制分析[D].博士学位论文.杨凌:西北农林科技大学, 2017. TIAN J J.Effects and mechanism analysis of arachidonic acid on lipid metabolism of grass carp (Ctenopharygodon idellus)[D].Ph.D.Thesis.Yangling:Northwest A&F University, 2017.(in Chinese)
[52] LI B, ZHANG X X, GUO F, et al.Characterization of tetracycline resistant bacterial community in saline activated sludge using batch stress incubation with high-throughput sequencing analysis[J].Water Research, 2013, 47(13):4207-4216.  
[53] 李秀玲, 刘宝锁, 刘波, 等.不同脂肪源对卵形鲳鲹肠道微生物菌群的影响[J].水产科学, 2020, 39(1):88-95. LI X L, LIU B S, LIU B, et al.Effects of different lipid sources on intestinal microbial flora in golden pompano Trachinotus ovatus[J].Fisheries Science, 2020, 39(1):88-95.(in Chinese)
[54] XU G L, XING W, LI T L, et al.Comparative study on the effects of different feeding habits and diets on intestinal microbiota in Acipenser baeri Brandt and Huso huso[J].BMC Microbiology, 2019, 19(1):297.
[55] 阮瑞, 吴金平, 李营, 等.人工养殖下达氏鳇幼鱼肠道菌群组成分析[J].淡水渔业, 2018, 48(5):93-98. RUAN R, WU J P, LI Y, et al.Composition of intestinal microbiota in juvenile Huso dauricus under artificial rearing[J].Freshwater Fisheries, 2018, 48(5):93-98.(in Chinese)
[56] 邸军.鲟细菌性败血症病原学及其防治研究[D].博士学位论文.重庆:西南大学, 2020. DI J.Study on etiology and prevention technique of bacterial septicemia in sturgeons[D].Ph.D. Thesis.Chongqing:Southwest University, 2020.(in Chinese)
[57] LIU H, GUO X W, GOONERATNE R, et al.The gut microbiome and degradation enzyme activity of wild freshwater fishes influenced by their trophic levels[J].Scientific Reports, 2016, 6:24340.
Options
文章导航

/

〈 〉

    联系我们

  • 地址：北京市海淀区圆明园西路2号
    　　　中国农业大学动科动医大楼153室
  • 邮编：100193
  • 电话：010－62817823
  • 网址：http://www.chinajan.com
  • E-mail：cjan@chinajan.com
京公网安备 11010802041926号    京ICP备17050989号-2