English
聯系我們
網站地圖
郵箱
舊版回顧



科學家破解基孔肯雅病毒入侵機制

文章來源:微生物研究所   發布時間:2019-05-10  【字號:     】  

  59日,中國科學院高福團隊在《細胞》(Cell雜志上發表了題為Molecular Basis of Arthritogenic Alphavirus Receptor MXRA8 Binding to Chikungunya Virus Envelope Protein 的文章,首次從分子水平闡釋了基孔肯雅病毒囊膜表面E蛋白與其細胞受體MXRA8分子的相互作用機制,揭示了此類病毒入侵細胞的分子機制,為抗病毒藥物開發及新型疫苗設計提供了新靶點。

  基孔肯雅熱(Chikungunya fever, CHIKF)是一種由基孔肯雅病毒(Chikungunya virusCHIKV)引起,經蚊蟲叮咬傳播的急性發熱性傳染病。該病毒病過去主要分布在非洲、南亞、東南亞熱帶和亞熱帶地區。近年來,隨著全球變暖和現代交通工具等因素,該病毒疫情呈不斷暴發和蔓延的趨勢。2005年以來疫情在東南亞和南亞地區再度暴發,2007年疫情傳播至歐洲,2013年疫情開始在加勒比海地區、美國以及南美洲等地暴發流行,造成了數百萬人感染。人感染該病毒后可致急性或慢性的外周關節痛或關節炎,嚴重時致人死亡,目前尚無特異性的治療方法和疫苗,使得該病毒引起人們的廣泛關注,也是重要的全球性公共衛生問題之一。與該病毒親緣關系較近,同樣能引起關節炎的甲病毒還包括馬亞羅病毒(Mayaro virus)、羅斯河病毒(Ross River virus)、阿尼昂尼昂病毒(O' nyong- nyong virus)等。

  病毒侵入細胞的第一步依賴于病毒囊膜蛋白與宿主受體的相互作用。過去的研究認為,CHIKV等多種致關節炎甲病毒囊膜表面含有80個三聚體形式的刺突E蛋白,每個三聚體由三個E1/E2二聚體組成其中E1蛋白參與膜融合,E2蛋白參與受體的結合。近來研究報道表明,基質重塑相關蛋白8Matrix remodeling-associated protein 8MXRA8)分子是CHIKV等多種致關節炎甲病毒的受體。MXRA8是一種在上皮細胞、髓細胞及間充質干細胞表達的黏附分子,廣泛分布于形成軟骨、肌肉和骨骼的細胞表面。但是,MXRA8分子如何介導病毒入侵卻是未解之謎。

  高福團隊深入研究了MXRA8分子介導CHIKV的入侵機制。首先,他們利用結構生物學的技術和方法解析了小鼠MXRA8mMXRA8)的晶體結構,表明MXRA8分子胞外段由兩個免疫球蛋白(Ig)樣結構域構成,但其兩個Ig樣結構域呈現獨特的拓撲結構。結構域1D1)由兩個不連續的片段構成,而結構域2D2)插在了D1的兩個片段之間,這導致D1D2之間有兩個柔性鉸鏈(Hinge loop)連接,這與以往所有報道的含有兩個Ig樣結構域的蛋白都不同。因此,他們首次提出MXRA8是一種新型的Ig樣受體分子,并具有獨特的拓撲結構及結構域間組裝形式。

  為進一步闡明CHIKV E與受體MXRA8相互作用機制,研究團隊解析了人MXRA8hMXRA8)與CHIKV E復合物的晶體結構,他們發現MXRA8CHIKV E采用一種獨特的3:3的結合模式,MXRA8結合到病毒表面三聚體刺突蛋白兩個E蛋白單體間的“峽谷”中,形成非常緊密的結合模式。其中E1E2均參與結合,MXRA8的兩個結構域及鉸鏈區均與E1E2蛋白發生相互作用。隨后他們利用冷凍電鏡技術解析了人MXRA8CHIKV病毒樣顆粒的復合物結構,證明了MXRA8在病毒表面的結合模式跟晶體結構所觀察到的結合模式一致,并通過點突變及表面等離子共振(SPR)方法對結合關鍵氨基酸進行了驗證。該研究首次“看清”了基孔肯雅病毒和受體相互作用的分子模式,為一種新型的病毒-受體結合模式,同時糾正了過去對基孔肯雅病毒和受體相互作用的一些錯誤認識。

  MXRA8因其獨特的拓撲結構排布而與其他I型跨膜蛋白不同,其ND1結構域不是遠膜端結構域,而實為近膜端結構域。在復合物結構中,MXRA8D1結構域深深地插入到CHIKV三聚體刺突E蛋白的“峽谷”中,這就需要近膜端的頸部區有足夠的長度和柔性。

  通過一系列MXRA8莖部區截短體和病毒及細胞水平功能實驗發現,MXRA8中長達48個氨基酸的莖部區為病毒入侵所必需,其足夠長且具有柔性,被病毒利用作為受體入侵細胞。

  此項研究首次揭示了致關節炎甲病毒CHIKVMXRA8受體相互作用機制,證實了MXRA8是一種具有獨特的拓撲結構及結構域間組裝形式的新型Ig樣受體分子,并發現了此類致關節炎甲病毒和受體相互作用是一種新型的病毒-受體結合模式。這些研究結果為新型疫苗及廣譜中和抗體的研發提供了理論指導,為抗病毒藥物設計提供了新靶點。

  中科院北京生命科學研究院助理研究員宋豪及微生物所碩士生趙振楠、助理研究員柴彥為論文的并列第一作者,中科院院士高福和中科院天津工業生物技術研究所副研究員高峰為論文共同通訊作者。微生物所研究員齊建勛、施一、嚴景華及巴西FIOCRUZ基金會衛生技術發展中心主任Carlos Morel給予了大力支持。該研究得到科技部重點研發計劃、中科院戰略性先導科技專項項目、國家科技重大專項、國家自然科學基金以及中科院青促會、中國科協“青年人才托舉工程”等的經費支持。

  文章鏈接

 

1. 小鼠MXRA8的三維結構

 

2. CHIKV E與人MXRA8復合物結構

 

3. MXRA8莖部區對CHIKV入侵很關鍵

 

4. CHIKV利用MXRA8入侵細胞的分子機制模型




(責任編輯:葉瑞優)

附件:

專題推薦

相關新聞


© 1996 - 中國科學院 版權所有 京ICP備05002857號  京公網安備110402500047號  聯系我們

地址:北京市三里河路52號 郵編:100864

华夏彩票www.hx5577.com 29027.com | 566984.com | www.86ku.com | www.331109.com | www.826957.com | www.cai3466.com | 773085.com | www.670034.com | www.095992.com | www.781213.com | www.91852.com | 大赢家彩票网www.54400w.com | www.027057.com | www.155683.com | www.573173.com | www.im79.com | www.713619.com | www.46584.com | 111075.com | 040956.com | www.96311.cc | www.220457.com | www.44565.com | 089357.com | 128387.com | 324281.com | www.787408.com | www.75018.com | www.51099.cc | www.304416.com | www.54nw.com | www.960855.com | 766624.com | 033904.com | www.558635.com | www.592273.com | 007926.com | www.297322.com | www.x52.cc | www.653512.com | www.613918.com | www.567159.com | 950843.com | www.737786.com | www.x18.pw | www.12186.com | www.047062.com | www.430721.com | 947634.com | www.238160.com | www.388624.com | www.875232.com | 788332.com | www.024487.com | 536729.com | www.373892.com | www.sq84.com | www.986898.com | www.161761.com | 548996.com | www.pw88.cc | www.8738.biz | www.6623.pw | www.51749.cc | www.004563.com | www.875660.com | 857240.com | 974845.com | www.874318.com | www.149700.com | www.655662.com | www.6481.vip | www.323788.com | www.777348.com | www.wp88.cc | www.874877.com | www.457688.com | 984959.com | www.092435.com | www.46035.cc | www.942233.com | 137906.com | www.183553.com | www.b53.xyz | www.602313.com | www.987977.cc | 121166.com | www.lb81.com | www.382068.com | www.001388.com | www.711917.com | www.220558.com | 518933.com | www.55884.cc | 899783.com | www.414900.com | www.660254.com | www.751988.com | www.940448.com | www.809177.com | www.196216.com | www.558633.com | www.867275.com | www.ps08.com | www.80819.cc | 743198.com | 418706.com | 562655.com | 350093.com | www.243025.com | 236535.com | 096647.com | 500彩票www.50080z.com | 233464.com | www.521477.com | www.636508.com | www.4613.cc | www.53432.com | 9号彩票www.80767u.com | www.1905.site | www.302322.com | www.461693.com | www.120126.com | www.889.net | www.520795.com | www.538.biz | www.520816.com | 093667.com | www.0120.net | 445218.com | www.914098.com | www.679279.com | www.174959.com | www.ss67.cc | 幸运彩票www.73166f.com | www.899074.com | www.kd46.com | www.726685.com | www.660819.com | 269201.com | www.cp508.cc | www.8994.vip | www.995138.com | www.129817.com | 220499.com | www.975348.com | www.21pk.com | www.193033.com | www.640517.com | www.397283.com | www.391600.com | www.0718.bid | www.2336.bid | www.2hs.cc | www.113197.com | www.866539.com | www.018072.com | www.165006.com | www.012648.com | www.948325.com | 423813.com | www.858453.com | 587400.com | www.040350.com | www.414182.com | 968426.com | www.as03.com | www.cai4442.com | www.725909.com | www.fo31.com | www.132462.com | www.783165.com | www.951687.com | www.dn54.com | 973274.com | 994861.com | 775406.com | www.126420.com | www.948840.com | 518733.com | www.6op.com | 500彩票网www.896020.com | www.599270.com | www.709649.com | www.914813.com | 787654.com | 316422.com | www.266.com | www.32176.com | www.162529.com | www.45640.com | www.130717.com | www.927870.com | www.287172.com | www.8jz.com | www.201635.com | www.840538.com | www.3888.top | www.5578.wang | www.21673.cc | www.56226.com | www.204111.com | 630395.com | 686718.com | www.877298.com | www.327353.com | www.399161.com | www.13ot.com | www.7276.net | www.228442.com | www.579462.com | www.x69.website | 109677.com | 678998.com | www.884546.com | 931591.com | www.91293.com | www.597590.com | www.cp3118.com | www.7571.loan | www.480806.com | www.455228.com | www.877666.com | 162081.com | www.vx32.com | www.605219.com | www.153626.com | www.686916.cc | 94cz.com | www.2953.biz | www.062494.com | www.368909.com | www.932017.com | www.z81.com | www.47ws.com | www.894810.com | www.923949.com | 474537.com | www.ji17.com | www.cb32.com | www.5131.biz | www.540280.com | www.480043.com | 833296.com | 397460.com | 309902.com | 新世界www.616777a.com | www.403575.com | 938473.com | www.ur35.com | www.5381.vip | www.576927.com | www.623969.com | www.01686.cc | www.nl79.com | www.rj3.com | www.715965.com | www.603892.com | 954934.com | www.86002.cc | 557426.com | 078934.com | www.fk10.com | www.539222.com | www.516222.com | www.105181.com | www.80378.com | www.631631.com | 028473.com | aj07.com | www.173071.com | 438477.com | www.806013.com | 301898.com | www.539308.com | www.098761.com | www.396281.com | www.266288.com | 488967.com | 梦想彩票站www.43131m.com | 中彩网www.81233l.com | 270939.com | www.5911.cn | 901876.com | 13dh.com--www.13dh.com | www.226053.com | www.zw42.com | 开心彩票www.596112.com | www.2579.win | www.235098.com | www.857951.com | www.59952.cc | www.093738.com | 大赢家彩票平台www.226105.com | www.jv03.com | www.4kj.cc | www.vw45.com | www.163682.com | www.408112.com | www.013722.com | www.412363.com | www.331989.com | 68彩票www.68689i.com | www.176479.com | www.876524.com | www.933533.com | www.692676.com | www.466949.com | www.202676.com | 645299.com | www.1687.vip | www.20bx.com | www.980590.com | 08yz.com | 804178.com | 905105.com | www.yy4.com | www.481819.com | www.440800.com | www.39713.com | www.049199.com | www.290117.com | www.407358.com | www.021905.com | www.659485.com | 福彩www.16878r.com | 277038.com | 彩71www.032823.com | 304894.com | 大赢家彩票平台www.377591.com | www.bq63.com | www.376455.com | www.890552.com | www.756773.com | www.002121.com | 576804.com | www.9058.vip | 681228.cc | www.746179.com | www.99bj.com | www.153060.com | 290323.com | www.cp917.com | www.97pd.com | www.8889.cm | www.2479.cc | www.7335.in | www.3157.top | www.017.in | www.178819.com | www.257006.com | www.618184.com | 240340.com | www.56hd.com | www.435180.com | www.7859.site | www.540570.com | www.590653.com | 033805.com--667345.com | www.ty26.com | www.725603.com | www.52963.cc | 250896.com | www.62611.com | www.013971.com | www.456886.com | www.719311.com | www.a89.me | www.09yn.com | www.17171.cc | www.587847.com | www.902005.com | www.vh.com | www.4530.biz | www.9749.in | www.124292.com | www.222306.com | www.645572.com | www.135546.com | 377197.com | 089052.com | www.25235.cc | www.666425.com | 221068.com | www.ec.cc | www.26hm.com | www.7070.xyz | www.292288.com | www.107216.com | www.kb1.com | www.93316.com | www.92pt.com | www.zg23.com | www.1967.cn | www.106411.com | www.075091.com | www.358972.com | www.386408.com | www.jg20.com | www.25zt.com | www.736078.com | 好运彩吧www.098286.com | www.t56.in | www.09605.com | www.7981.net | www.93752.com | www.40636.com | www.2399.in | www.863758.com | www.78603.com | www.968966.com | www.128637.com | www.2xs.com | www.911888.cc | www.092230.com | 268973.com | www.493448.com | 亿万彩票www.770640.com | 055703.com | 红旗彩票www.895594.com | 636597.com | www.397081.com | www.369294.com | www.951318.com | www.vv26.com | www.tc01.cc | 139930.com | www.0582.me | www.93vs.com | www.836139.com | www.848451.com | www.cp1110.com | www.345189.com | www.252330.com | www.862852.com | 190572.com | www.14dr.com | www.25319.com | www.8049.loan | www.672.in | www.46889.com | www.136509.com | www.157715.com | www.790512.com | www.2018999.com | www.833991.com | 964839.com | www.029198.com | www.262157.com | 984632.com | 519525.com | www.wq78.com | www.og78.cc | www.d79.net | www.sr36.com | 780636.com | www.447270.com | www.848856.com | www.794163.com | www.az99.cc | www.4512.me | www.203795.com | www.819795.com | www.3529.win | www.311.cn | www.kg09.com | www.82dw.com | www.03cs.com | www.9896.cm | www.65411.cc | www.598078.com | www.708119.com | www.278365.com | www.233743.com | www.240110.com | www.500177.cc | www.534336.com | www.426992.com | www.026759.com | www.033499.com | www.975804.com | www.944945.com | 488965.com | www.44200.cc | www.37282.cc | www.343709.com | 彩民村www.88cmc.com | 671496.com | www.971616.com | www.466189.com | 99彩www.506586.com | www.32mw.com | www.21ey.com | www.652338.com | 500彩票www.50024v.com | www.75lt.com | www.9851.in | www.8830.vip | www.55702.com | www.150626.com | www.75395.cc | www.779376.com | www.16co.com | www.pd67.com | www.0906.mobi | 大赢家彩票www.799343.com | 796291.com | www.65203.com | www.267122.com | www.276250.com | www.634009.com | www.81gi.com | www.21ha.com | www.560073.com | 967062.com | 581190.com | www.81zw.com | www.52353.com | www.051584.com | www.379238.com | www.744252.com | www.467739.com | www.781336.com | www.971509.com | 14pj.cc | 659675.com | www.027541.com | 961642.com | www.832227.com | www.332030.com | www.222711.com | www.73rc.com | www.528110.com | www.776793.com | www.644444.com | 429783.com | www.16ma.com | www.176582.com | www.714422.com | www.210978.com | www.658041.com | www.545453.com | www.557170.com | www.473783.com | www.491345.com | www.403575.com | www.628250.com | 466859.com | www.042223.com | www.290829.com | www.997429.com | www.886374.com | www.786555.com | www.53126.cc | www.83de.com | www.243568.com | www.tf66.com | 656888.com | www.952.tv | www.127376.com | www.99293.cc | www.42826.com | www.010090.com | www.71ah.com | www.924011.com | www.1268.xyz | www.34777.cc | www.01661.com | www.85883.cc | www.55210.com | 大赢家彩票www.399049.com | www.17898.com | www.106041.com | www.725691.com | www.595125.com | www.334164.com | 894163.com | www.fc11.cc | www.2399.vip | www.142.mobi | www.962555.com | www.904880.com | www.904536.com | 855883.com | www.591234.com | 94lg.com | 七星彩票www.015689.com | www.g32.net | www.206560.com | www.68244.com | www.25me.com | by77.cc | 166237.com | www.745754.com | www.659947.com | www.907106.com | www.zh3.cc | 00ak.com | www.fy61.com | 杏彩www.00840b.com | www.nk05.com | www.la19.com | www.001290.com | www.1171.in | www.109408.com | 012567.com | 044182.com | 557597.com | www.33896.com | www.548864.com | www.052360.com | www.708933.com | www.639729.com | www.063797.com | www.672596.com | www.323788.com | 965742.com | 407256.com | www.cai24.cc | www.21246.com | www.808599.com | www.971390.com | 796399.com | 778601.com | 534357.com | www.57808.com | www.681535.com | www.598553.com | www.42654.com | www.79614.com | www.121081.com | www.52681.com | www.48166.com | www.46uf.com | www.17462.com | www.757588.cc | www.448229.com | www.822027.com | www.126111.com | www.706033.com | www.962501.com | www.mi.com | 964284.com | www.900166.com | www.64407.com | www.220396.com | www.106896.com | www.365626.cc | www.791886.com | 239268.com | www.996100.com | 彩61www.649638.com | www.363527.com | www.029807.com | www.367028.com | www.664686.com | www.877500.com | 168381.com | 08450.com | www.0rr.com | www.oa27.com | www.ts54.com | www.ta35.com | www.258584.com | www.234340.com | www.867638.com | www.152.in | www.050.cx | 086049.com | www.348360.com | www.34359.com | www.971500.com | www.008054.com | www.696793.com | www.608569.com | www.446548.com | www.380.cm | www.94yc.com | 694978.com | 500彩票www.653182.com | 690106.com | www.63330.cc | www.554015.com | www.751959.com | www.394847.com | www.197449.com | www.333353.com | www.jn59.com | 945552.com | www.la14.com | www.3824.wang | www.31318.cc | www.870.gg | www.73kl.com | www.664.la | www.95062.com | www.292991.com | www.769399.com | www.cp9552.com | 627732.com | www.385822.com | www.900966.cc | www.853641.com | 774414.com | 9643.biz | 487011.com--200444.com | www.710646.com | 950680.com | www.972717.com | 000049.cc | www.22133.com | www.150959.com | www.939335.com | www.603704.com | www.327699.com | www.597223.com | www.558199.com | www.512572.com | www.41293.com | www.874286.com | www.03971.com | www.259.in | www.6845.cn | www.0999.cc | www.294618.com | www.964504.com | 227012.com | www.ao88.cc | www.766623.com | www.354835.com | www.378859.com | www.135209.com | www.171029.com | www.586026.com | www.ci22.com | www.52198.cc | www.394100.com | www.818225.com | www.473111.com | www.281237.com | www.365145.cc | www.628090.com | 519478.com | 941707.com | www.781103.com | 974203.com | 984953.com | www.xpj.com | www.3598.xyz | www.313195.com | www.j28.cn | www.24nk.com | www.47vd.com | www.8189.pw | www.502289.com | www.139720.com | www.247904.com | www.364234.com | www.659364.com | 奔驰彩票www.281877.com | 8146.biz | www.746224.com | www.969804.com | www.891466.com | www.054665.com | 优彩网www.974091.com | www.505274.com | www.825398.com | www.533923.com | www.964712.com | www.bk29.com | www.93316.cc | www.591855.com | www.87916.com | 中彩网www.61233k.com | www.6789789c.com | 731864.com | www.wc08.com | www.93073.com | www.aw12.com | 329080.com | 936117.com | 530649.cc | 780198.com | www.24388.com | www.36nu.com | 324711.com | 679103.com | www.dw7.com | www.570668.com | 505298.com | 085579.com--163a.com | www.956049.com | www.770391.com | 878075.com | www.035725.com | cai0002.com | 085965.com | www.206912.com | www.12ez.com | 747295.com | www.865617.com | www.524408.com | www.774598.com | www.458095.com | 533844.com | www.260172.com | www.53381.com | 幸运彩票www.5095t.com | www.iz21.com | 326616.com | www.03jn.com | www.cai3459.com | 686888.com | 954650.com | www.009832.com | 042461.com | www.804888.com | www.599001.com | www.662758.com | 520195.com | www.007959.com | www.33136.com | www.587608.com | www.807861.com | www.258948.com | www.288438.com | www.505697.com | www.909271.com | 158710.com | 358714.com | 139462.com | 614613.com | www.317011.com | www.962935.com | 434438.com | 292873.com | www.wr2.com | www.24.gg | www.6849.vip | www.761799.com | 035075.com | www.73225.cc | www.019477.com | www.42mk.com | www.vg95.com | www.603329.com | www.050052.com | www.51740.com | www.12317.com | www.917006.com | www.533716.com | www.026316.com | www.532958.com | www.183172.com | www.257526.com | www.036621.com | www.032550.com | www.5115.pw | 1js.com | www.620049.com | www.283082.com | www.836803.com | 3181.biz | 926260.com | www.062608.com | www.rj13.com | 500彩票www.645977.com | 博亿彩票www.622by.com | www.18715.com | www.78223.cc | www.0651.in | 62np.com | www.889557.com | 285966.com | www.468578.com | www.oq74.com | www.628009.com | www.8665.club | www.798257.com | 036334.com | www.16281.cc | www.610679.com | www.255893.com | www.95792.com | www.5518.cc | www.51dx.com | www.vh39.com | 029369.com | 722268.com | www.947164.com | 彩69www.095335.com | www.784208.com | www.893991.com | www.523156.com | www.113351.com | www.080835.com | www.050200.com | www.3309.tv | www.505.cm | www.48yy.com | 福彩网www.055123.cc | 111981.com | www.331456.com | www.417855.com | www.948230.com | www.605363.com | 702198.com | 321478.com | www.72687.com | 004514.com | 037568.com | 635663.com | 890525.com | www.813524.com | www.500088.com | www.1311.vip | www.718651.com | www.099111.com | www.2923.xyz | 4377.vip | www.978600.com | www.702111.cc | www.296573.com | www.206496.com | www.33033.com | www.tb82.com | 291175.com | www.3452.in | www.670292.com | www.690052.com | www.cj5500.com | www.222764.com | www.753298.com | www.415108.com | www.206033.com | www.010995.com | www.18846.com | www.08jb.com | www.150603.com | www.1502.me | www.46.me | 500彩票www.574177.com | www.02ta.com | www.2189.org | www.140422.com | www.581098.com | www.84447.cc | www.414196.com | 明星彩票www.cp67.cc | www.9971.cn | 幸运彩票www.luck878.com | 613729.com | www.896185.com | www.009208.com | cai2224.com | www.879032.com | www.476747.com | www.039693.com | www.4727.biz | 中彩网www.81233v.com | 88734.com | 001219.com | www.798681.com | www.441886.com | www.064469.com | www.05781.com | www.1000.biz | www.68655.com | www.39193.com | www.u29.club | www.c10.top | 675910.com | www.113092.com | www.043195.com | www.391203.com | www.504600.com | www.ve83.com | 338264.com | www.859.cm | www.536.cn | www.12lu.com | www.17796.com | www.b62.club | 793101.com | 977961.com | www.79494.com | www.787577.com | www.879326.com | 972055.com | www.172785.com | www.878337.cc | www.948369.com | www.878742.com | www.788904.com | www.820148.com | 024875.com | www.2067.vip | 655457.com | www.867757.com | www.631036.com | 262222.com | www.17uc.com | www.oy42.com | www.632019.com | 彩票大家乐www.21202e.com | 3523.com | www.917159.com | www.413880.com | www.yp6.cc | www.888677.com | www.637398.com | www.624533.com | www.894416.com | www.531154.com | www.261594.com | 947489.com | www.840811.com | www.366627.com | www.45vc.com | www.284156.com | www.220572.com | 736171.com | www.829197.com | www.ks02.cc | www.557733.com | www.316785.com | www.092630.com | www.893150.com | www.559787.com | www.537013.com | www.6261.biz | www.81867.cc | www.385101.com | www.379773.com | www.823271.com | www.95pj.cc | www.837057.com | 84411.cc | www.771017.com | www.200.xyz | www.91472.com | www.046589.com | www.22959.cc | www.66927.cc | www.89bg.com | www.jt15.com | www.cai3344.com | www.ug16.com | 463412.com | 409556.com | www.213548.com | 624949.com | www.179114.com | www.311604.com | www.201591.com | www.563861.com | 843322.com | www.928250.com | www.893095.com | www.362614.com | www.998281.com | www.880502.com | www.728355.com | www.297632.com | www.5284.net | www.342655.com | ke30.com--ke30.com | www.83822.cc | www.391659.com | www.85743.com | www.1059.cc | 967470.com | www.13529.cc | www.72ma.com | 中彩网www.71233k.com | www.x55.top | www.8884.loan | www.35iz.com | www.036317.com | www.9672.me | www.8674.com | cr91.cc | 127460.com | www.921223.com | www.630280.com | www.721969.com | www.800070.cc | www.769484.com | www.910992.com | www.697629.com | www.053785.com | 4157.biz | www.855379.com | 830389.com | 181066.com | 2072.com | 43xj.com | www.173632.com | www.1357.xyz | www.11729.com | www.34.cc | www.634230.com | www.qx3.cc | www.964637.com | www.556000.com | www.576381.com | www.239519.com | www.98uq.com | 111656.com | www.11yg.com | 882404.com | www.380923.com | www.085208.com | www.129437.com | www.093699.com | www.70551.com | www.zl6.cc | www.197.xyz | www.425214.com | www.3459.net | www.029486.com | www.qx21.com | www.mw01.com | www.lz88.com | www.81aw.com | 博通彩票www.694050.com | www.939193.com | www.374144.com | www.769602.com | www.506893.com | www.536.website | www.388981.com | www.562609.com | www.462216.com | www.127086.com | 920061.com | www.1855.live | www.hf49.com | 236157.com | www.595331.com | 074386.com | 091081.com | www.cp5600.com | 517705.com | 500彩票www.50051t.com | 739339.com | www.872451.com | 519930.com | www.5320.cn | www.hh62.cc | www.28911.com | www.av27.com | 424889.com | www.581753.com | www.44wn.com | www.678.net | www.053477.com | 215351.com | 971385.com | www.775533.com | www.916205.com | www.954198.com | www.508244.com | www.150169.com | www.632652.com | 895634.com | www.970589.com | www.678763.com | www.427846.com | www.16959.com | www.34kn.com | www.72ct.com | www.420153.com | www.820977.com | www.283555.com | www.98875.com | www.5du.cc | 008034.com | 985458.com | www.91dz.cc | www.i65.com | www.lm0.cc | www.tf57.com | www.028641.com | www.729279.com | www.506368.com | 284921.com | www.826276.cc | www.600090.cc | 084661.com | www.25hb.com | www.341185.com | www.591522.com | www.543442.com | www.954007.com | www.960846.com | www.iv08.com | 388713.com | www.983896.com | www.703968.com | www.333713.com | www.6633.vip | 639646.com | www.907909.com | www.293366.com | www.3767.xyz | 293468.com | 456141.com | www.540420.com | www.81854.com | www.081042.com | www.2372.biz | bm30.com | www.983335.com | www.535397.com | www.903030.com | www.571612.com | www.255369.com | www.688764.com | www.867373.com | www.238258.com | www.803189.com | www.222516.com | www.294832.com | www.4397.biz | 彩82www.08992.com | www.941252.com | www.343727.com | 867056.com | www.924833.com | www.530082.com | www.653374.com | 062886.com | www.795857.com | www.543166.com | www.917099.com | 707526.com | www.729456.com | www.185845.com | www.34732.com | www.0845.org | 552642.com | www.950673.com | www.335368.com | www.8822.online | 848698.com | 22828.cc | www.662083.com | www.184561.com | www.68043.com | www.96vg.com | 211447.com | www.645207.com | www.400122.com | www.619999.com | www.033617.com | www.185.date | 971443.com | www.838394.com | www.393138.com | www.86022.com | www.db08.com | 891245.com | www.ng11.cc | www.169010.com | www.125086.com | www.e26.org | www.996351.com | www.384919.com | www.030015.com | www.930583.com | www.659116.com | www.916994.com | www.717214.com | www.853767.com | www.124838.com | www.1313.vip | www.bw80.com | 367206.com | 127392.com | www.690000.com | www.900411.com | www.777354.com | www.396500.com | www.440111.com | www.78285.cc | www.75jm.com | www.136303.com | www.72778.cc | www.85wu.com | www.99bc.cc | www.tg33.com | 260744.com | www.962128.cc | 亿彩彩票www.ycp5.com | www.899711.com | www.305598.com | www.8483.cc | 百姓彩票www.www.bxcp3333.com | 582167.com | www.783995.com | www.587445.com | www.288385.com | www.036523.com | www.w50.xyz | 456231.com | www.729411.com | www.210363.com | www.3505.cm | 377089.com | www.931120.com | www.408679.com | www.075507.com | www.11ue.com | www.960686.com | www.253944.com | 234668.cc | www.920233.com | www.227111.cc | www.27ag.com | www.996709.com | www.707027.com | www.65377.cc | www.2co.com | 816495.com | www.684479.com | www.162079.com | www.394400.com | www.440162.com | www.8084.cn | 网易彩票www.07163y.com | 018673.com | www.19qq.cc | 739486.com | www.678110.com | www.r91.club | 532573.com | www.5952.win | www.cp770.com | www.gg22.cc | 1409.cc | www.876584.com | www.732785.com | www.984703.com | www.622099.com | www.560534.com | www.906031.com | www.994423.com | www.610351.com | www.511478.com | www.117946.com | www.5533.live | www.579484.com | www.177396.com | 628675.com | www.618433.com | www.230876.com | www.367.tv | www.b97.club | 9583.biz | www.86zf.com | www.gl1.cc | 661409.com | www.762275.com | www.284172.com | www.092241.com | www.34ub.com | 964810.com | www.948413.com | www.451555.com | www.744464.com | www.972307.com | www.515650.com | www.594040.com | www.439833.com | www.002543.cc | www.211242.com | www.583652.com | www.157309.com | www.161071.com | www.123397.com | www.6498.vip | 969426.com | www.980586.com | 331649.com | www.557911.com | www.235470.com | www.020769.com | www.30di.com | 1095.xyz | www.857393.com | www.533910.com | www.108098.com | 557556.com | 097829.com | www.948693.com | www.590622.com | www.085331.com | www.83538.com | www.2493.com | www.133937.com | www.75650.cc | www.98rd.com | 爱彩www.014ac.com | www.954460.com | www.994739.com | www.515865.com | www.062476.com | www.316439.com | www.428559.com | www.250003.com | www.3567.cc | 482609.com | www.914022.com | www.482061.com | www.065636.com | www.59pm.com | 826023.com | www.786981.com | www.285676.com | www.995077.com | www.542814.com | www.66591.com | www.368.bid | 541166.com | www.937996.com | www.835862.com | www.346349.com | www.3084.biz | 460037.com | www.853694.com | www.382295.com | www.204078.com | www.2714.me | 彩票坊www.022cq.cc | www.957201.com | www.429608.com | www.739976.com | www.690157.com | www.232799.com | www.22571.cc | www.1840.org | 714122.com | 796514.com | www.s06.org | www.299817.com | www.588001.com | www.cp996.com | www.291520.com | 722349.com | www.93650.com | www.960546.com | 029970.com | 241601.com | www.719849.com | www.541005.com | 彩61www.097968.com | www.301355.com | 207169.com | www.342348.com | www.612555.com | www.223746.com | www.6sc.com | www.6816.biz | 980586.com | www.805057.com | www.214389.com | www.177864.com | www.39764.com | www.ld35.com | www.81da.com | 506002.com | www.947536.com | www.634376.com | www.369493.com | www.058138.com | www.081089.com | www.5438.in | 494558.com | 31399.com | www.773164.com | www.430056.com | www.156681.com | www.323657.com | www.312330.com | www.002366.com | www.dg96.com | 267789.com | www.878849.com | www.346982.com | www.226044.com | www.23143.com | 天天赢彩票www.73736f.com | 052887.com | 981757.com | www.893268.com | www.265250.com | www.036032.com | www.w75.org | 482168.com | 67324.cc | www.961815.com | www.872940.com | www.639034.com | www.496318.com | www.712983.com | www.053583.com | www.td26.com | www.907711.com | www.593028.com | www.081856.com | www.21633.cc | www.a50.wang | www.989926.com | www.355709.com | www.087212.com | www.04wf.com | 122994.com | www.787852.com | www.645246.com | www.192567.com | www.449832.com | www.756888.cc | www.417666.com | www.197803.com | www.1134.site | www.fl20.com | 057810.com | www.607223.com | www.313636.com | www.313618.com | www.083803.com | www.1080.pro | yl69.cc | www.948839.com | www.771163.com | www.559580.com | www.318641.com | www.595156.com | www.082046.com | www.xs96.com | www.47fp.com | www.76fn.com | www.195.la | www.wp39.com | www.13hn.com | www.973594.com | www.377108.com | www.87665.cc | www.i59.biz | 020827.com | www.791664.com | www.428878.com | www.11879.com | www.zy68.com | www.830624.com | www.224755.com | www.28225.cc | www.3085.vip | 461489.com | www.903801.com | www.328674.com | www.7799.vip | www.23272.cc | 103643.com | www.764803.com | www.791819.com | www.309005.com | www.02768.com | 易彩网www.12455l.com | www.847858.com | www.795863.com | www.266288.com | www.784567.com | www.185728.com | www.041078.com | www.206296.com | www.0711.net | JJ彩票www.12018.cc | 849820.com | www.870812.com | www.714571.com | www.670070.com | 102890.com | www.945199.com | www.559661.com | www.