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

金冠彩票www.c4534.com 530944.com | www.55682.com | www.209751.com | www.54411.cc | www.315748.com | www.377903.com | www.963917.com | www.n91.net | 190445.com | www.89579.com | www.118003.com | www.948985.com | 110761.com | www.98sn.com | www.165053.com | www.102036.com | cp617.com | www.277014.com | www.447349.com | www.808493.com | www.929373.com | www.fp03.com | www.079156.com | www.191591.com | 汇彩网www.www.hcw266.com | www.567284.com | 964226.com | 776657.com | www.707979.com | www.cai9077.com | 849646.com | www.320608.com | 949851.com | 091540.com | www.cp8638.com | www.01ik.com | www.070778.com | www.380806.com | www.591208.com | www.p66.top | www.17429.com | www.065065.cc | www.449590.com | 979449.com | www.51568.com | www.001594.com | www.303237.com | www.482868.com | www.jo87.com | www.33he.com | www.572101.com | www.62ub.com | www.333644.com | 824931.com | www.839158.com | 581858.com | 923153.com | www.771225.com | www.188160.com | 589550.com | www.ek40.com | 天下彩www.854980.com | www.co41.com | www.7709.vip | www.2kk.com | www.616448.com | 500彩票网www.61655w.com | www.k10.shop | 872824.com | www.774101.com | www.97yi.cc | www.5984.org | www.077789.com | www.10169.com | www.102080.cc | www.170370.com | www.512190.com | 幸运彩票www.717699.com | www.ol7.com | www.661041.com | 692790.com | 787971.com | 502196.com | www.fh47.com | www.6655.loan | www.64424.com | 687282.com | www.16457.com | www.46661.com | www.046011.com | www.065807.com | www.28851.com | www.759135.com | www.175611.com | 020679.com | www.oq27.com | www.197031.com | 105489.com | 087356.com | 789787.com | www.2883.biz | 757904.com | 网易彩票www.069wy.com | 105448.com | www.5913.me | www.154446.com | www.922535.com | www.863092.com | www.536044.com | www.271909.com | www.218723.com | www.769713.com | www.16543.com | 879234.com | 757994.com | www.98rs.com | www.sf33.cc | www.308301.com | www.30694.com | www.423326.com | www.380029.com | www.654271.com | www.lc64.com | www.yy01.cc | www.29133.com | www.ri82.com | www.c47.link | 588281.com | 365149.cc | 500彩票www.377501.com | 872602.com | www.733342.com | www.86dt.com | www.09291.com | www.6513.biz | www.40103.com | www.207.cm | www.950297.com | www.256978.com | www.ea72.com | www.393277.com | 374360.com | www.562853.com | www.590363.com | www.650237.com | www.600762.com | www.982473.com | www.22oq.com | www.94733.com | www.028490.com | www.q38.org | www.802282.com | 747999.com | www.gs1.com | 大赢家彩票平台www.226065.com | 139147.com | www.445515.com | www.720663.com | 790635.com | www.6412.vip | www.028218.com | www.970891.com | 计划网页版 | www.108579.com | www.926516.com | www.962743.com | www.5737.net | www.886957.com | www.yi22.com | www.053290.com | www.247244.com | www.75578.com | www.957395.com | 631880.com | www.995447.com | www.977764.com | www.250481.com | www.9520.vip | www.67id.com | 724013.com | www.104117.com | www.4hk.cc | www.773085.com | www.zr39.com | www.85619.cc | www.184228.com | www.05419.com | www.639624.com | 538235.com | 030041.com | www.51aa.com | www.89549.com | 222065.com | www.88297.cc | www.jk46.com | www.669479.com | www.41ux.com | www.029680.com | www.570022.com | 070765.com | www.059566.com | www.161386.com | www.596359.com | www.253846.com | www.et9.com | www.26lb.com | www.398025.com | www.684672.com | iv80.com | www.zq71.com | www.59936.cc | www.175626.com | www.620137.com | www.303016.com | www.464544.com | www.373239.com | www.174719.com | www.673840.com | www.80lw.com | www.7830.pw | www.838.cm | www.218.red | www.206263.com | www.78460.com | www.656365.cc | www.970468.com | www.xj23.cc | www.gd09.com | www.287580.com | www.183510.com | www.338656.com | www.175325.com | 111564.com | 611755.com | www.v46.net | www.743255.com | www.680775.com | www.502198.com | www.588317.com | 幸运彩票www.444957.com | www.ae21.com | www.zg74.com | www.76855.com | 789838.com | www.814011.com | www.073094.cc | www.3382.pw | www.391300.com | www.85ja.com | www.dt56.com | www.4748.cc | www.986545.com | 040410.com | www.dm40.com | www.33ik.com | www.025682.com | www.98128.cc | www.9061.com | www.141199.com | www.026585.com | www.062327.com | www.391679.com | www.510613.com | www.639646.com | www.937655.com | 954861.com | www.74zm.com | www.230767.com | www.378100.com | www.393888.com | www.771222.com | www.434434.com | www.581210.com | www.680751.com | www.987617.com | www.854945.com | www.585265.com | www.761708.com | www.851926.com | www.881413.com | www.141991.com | www.282100.com | www.201538.com | www.91047.com | www.022196.com | www.820198.com | 188743.com--88msc.com | www.779851.com | www.mx16.com | www.52hc.com | www.co57.com | www.oh95.com | www.379176.com | www.865.red | www.026272.com | www.145679.com | www.768108.com | www.691415.com | www.700225.com | www.865313.com | 03nu.com | www.880877.cc | 111605.com | www.773551.com | www.827289.com | 365624.cc | www.739472.com | www.202422.com | www.285171.com | www.994348.com | www.952503.com | www.973397.com | www.956150.com | www.17nl.com | www.9tz.cc | www.100.com | www.6868.com | 829079.com | www.gg2.com | www.537508.com | www.522382.com | 利彩www.57578b.com | 947167.com | 595026.com | www.94nj.com | www.439943.com | www.940098.com | 94338.com | www.134821.com | www.ca6.com | www.cy15.com | 836995.com | www.547326.com | www.552327.com | www.513711.com | www.507836.com | www.458199.com | 896772.com | www.g50.top | www.1xw.com | www.24ul.com | www.3805.cm | www.89065.com | www.101835.com | www.062527.com | www.761881.com | www.792565.com | www.762352.com | www.208606.com | www.cp776.com | www.537172.com | www.599234.com | 438765.com | www.042597.com | www.25066.com | www.253878.com | www.237000.cc | 501368.com | www.308026.com | www.668858.cc | 480300.com | www.us62.com | www.702676.com | www.7113.net | www.914033.com | 2995.pw | www.187621.com | www.049055.com | www.86387.cc | www.150494.com | www.238339.com | www.254061.com | www.62422.com | www.992346.com | www.806297.com | www.j95.in | www.916386.com | 464529.com | www.702227.com | www.918814.com | www.374755.com | www.438866.com | www.654003.com | www.xpj88.com | www.504244.com | www.636080.com | www.mh52.com | 845200.com | 905568.com | 中彩网www.71233w.com | www.cai8887.com | www.520335.com | www.458722.com | www.631623.com | 118268.com | www.in99.com | www.389913.com | www.64rd.com | www.058818.com | www.300644.com | www.542244.com | www.393978.com | www.1301.org | www.9647.net | www.753672.com | www.390.in | www.17866.cc | www.765262.com | www.526234.com | www.77103.com | www.029873.com | www.991729.com | www.ce4.com | www.16446.com | www.029377.com | www.386308.com | 635957.com | 974962.com | www.089095.com | 554613.com | www.wb42.com | www.es61.com | www.fk72.com | 474810.com | www.655367.com | www.612708.com | www.579574.com | www.604669.com | www.562923.com | www.543648.com | www.540730.com | www.499573.com | www.371360.com | www.329474.com | www.239613.com | www.150607.com | www.136247.com | www.194544.com | www.188085.com | www.197809.com | www.134031.com | www.681572.com | www.330823.com | www.940047.cc | www.965789.cc | www.16mf.com | www.e91.com | www.3ba.cc | www.xl33.cc | www.hq47.com | 13e彩票www.809381.com | 368448.com | 522583.com | 367569.com | 190393.com | 458498.com | 242233.com | www.919715.com | 222089.com | 900358.com | 409166.com | www.925025.com | www.817277.com | www.726708.com | www.655766.com | www.589404.com | www.530612.com | www.507921.com | www.548986.com | www.441194.com | www.321321.com | www.351445.com | www.278678.com | www.222619.com | www.156497.com | www.179732.com | www.127929.com | www.70789.com | www.35166.com | www.6776.top | www.3154.live | www.99fp.com | www.oa45.com | www.ab38.com | 563588.cc | 0127.com | cp739.com | 30419.com | 017832.com | www.982510.com | www.936685.com | www.831094.com | www.728083.com | www.657619.com | www.554999.com | www.493910.com | www.348422.com | www.267297.com | www.186952.com | www.121916.com | www.206854.com | www.096371.com | www.004442.com | www.32126.cc | www.2426.org | www.34sr.com | www.tq57.com | 幸运彩票www.444539.com | 487920.com | 008496.com | 202836.com | www.966730.com | www.872305.com | www.766368.com | www.491574.com | www.380202.com | www.265779.com | www.191729.com | www.096331.com | www.007243.com | www.13807.cc | www.2425.net | www.13sg.com | www.ik01.com | 873710.com | 820568.com | 277058.com | 456819.com | www.975029.com | www.879418.com | www.927025.com | www.024851.com | www.963407.com | www.855821.com | www.715283.com | www.594228.com | www.351946.com | www.026708.com | www.26hk.com | 213823.com | www.3862.cc | www.65nq.com | www.tg14.com | 500彩票www.50066c.com | 311058.com | 694234.com | 394480.com | www.970962.com | www.947183.com | www.347311.com | www.22331.cc | www.974953.com | www.943603.com | www.934504.com | www.731413.com | www.771566.com | www.831040.com | www.547730.com | www.984019.com | www.717508.com | www.370939.com | www.038448.com | www.3127.pw | www.qe76.com | 385207.com | 605499.com | www.544099.com | 973148.com | www.pp42.com | www.5206.vip | www.102306.com | www.131118.com | www.695920.com | www.zr75.com | www.558281.com | www.874402.com | www.059667.com | www.55481.com | www.276378.com | www.191008.com | www.926385.com | www.929667.com | www.956505.com | 938268.com | www.345388.cc | www.6876.org | 彩69www.214656.com | www.q88.website | www.31yk.com | www.w62.net | www.06549.com | www.202630.com | www.125403.com | www.mn17.com | www.513711.com | www.442666.com | www.447966.com | www.422787.com | www.397311.com | www.904948.com | www.082232.com | www.g67.com | 凤凰彩票www.66652c.com | www.7354.tv | www.19272.com | www.am35.cc | 快彩www.81678l.com | www.26815.cc | www.3311.vip | www.20032.com | www.642042.com | www.464046.com | www.345183.com | www.238678.com | www.219218.com | www.590813.com | 168005.com | www.lk66.com | www.79553.cc | www.ix6.com | www.me35.com | www.499049.com | www.885035.com | www.w83.club | www.1615.vip | www.635954.com | www.611206.com | 大玩家彩票www.315019.com | www.266071.com | www.923273.com | www.727279.com | www.gz.cc | 381205.com | 254393.com | www.021293.com | www.430582.com | www.239533.com | www.773812.com | www.062436.com | www.52778.com | www.111136.com | www.098033.com | www.98156.com | 408146.com | 快彩在线www.cpkk6.com | www.0214.online | www.032046.com | www.512963.com | 15574.com | 905178.com | www.828969.com | www.878165.com | www.730578.com | www.590490.com | www.635689.com | www.587786.com | www.812861.com | www.508.vip | www.96bk.com | www.523390.com | www.835554.com | 366453.com | www.339155.com | www.566138.com | www.999748.com | www.98820.cc | www.198491.com | www.3773.com | 358472.com | www.la09.com | www.085305.com | www.110697.com | 027904.com | tm9.cc | 144915.com | 947747.com | 338674.com | www.of9.com | 292769.com | www.256005.com | www.350642.com | www.588627.com | www.027636.com | www.bb27.com | www.516767.com | www.07326.com | www.1464.xyz | www.c64.net | 中彩网www.81233cc.com | 226836.com | 190699.com | 97by.cc | 892769.com | www.992629.com | www.820727.com | www.691242.com | www.500555.cc | www.301664.com | www.122575.com | www.70033.com | www.5447.cc | www.57gb.com | www.dt70.com | 233184.com | www.gl4.com | www.067711.com | www.139020.cc | 67550.com | www.982854.com | 943543.com | 708192.com | 582352.com | www.213566.com | www.904128.com | www.501296.com | www.450504.com | 177844.com | www.856255.com | eb90.com | 183967.com | 必发彩票www.925507.com | www.128659.com | www.966862.com | www.796271.com | www.981745.com | www.664678.com | www.512644.com | www.080062.com | www.274625.com | www.33136.com | www.034960.com | www.04297.com | www.zx85.com | www.ox71.com | www.945556.com | www.672008.com | www.737029.com | www.557862.com | www.725308.com | www.427058.com | www.113349.com | www.039254.com | www.7287.cn | www.64uk.com | 250867.com | www.li61.com | www.ku0.cc | www.18078.com | www.771415.com | www.667281.com | www.050704.com | www.965834.com | www.445572.com | www.877068.com | www.974861.com | 212287.com | www.881457.com | xwcp.com | www.966801.com | www.386763.com | www.509839.com | 55233.cc | www.389157.com | www.380582.com | www.97qh.com | www.sk90.com | 531510.com | www.xpj432.com | www.952697.com | www.678337.com | www.486353.com | www.183773.com | www.77312.cc | www.21jy.com | www.42sn.com | www.cp470.cc | www.691973.com | www.242585.com | www.122961.com | www.ly96.com | www.85ki.com | www.789094.com | 378948.com | 301648.com | cai1239.com | 111043.com | www.606923.com | www.942097.com | www.903638.com | www.99ka.com | www.501982.com | www.117249.com | www.560466.com | www.298779.com | www.356693.com | 241115.com | www.119464.com | www.44877.cc | www.cp799.com | 591716.com | 018939.com | www.978416.com | www.22fy.com | www.lr40.com | 562607.com | www.5561.cc | 5865.biz | 812392.com | 848785.com | 811035.com--163a.com | www.899116.com | www.685057.com | www.377531.com | www.177024.com | www.82511.cc | www.74641.cc | www.0476.love | www.co82.com | 168024.com | www.973409.com | www.705656.com | www.388366.com | www.259058.com | www.040094.com | www.9559.cc | www.45xz.com | www.837844.com | 055331.com | www.858425.com | www.583120.com | www.322400.com | www.112029.com | www.19456.cc | www.7kt.cc | 818合彩www.www.8000hc.com | 462085.com | www.956083.com | www.750988.com | www.615468.com | www.379333.com | www.148050.com | www.41769.com | www.0220.net | www.aa57.com | www.39499.com | www.890834.com | www.356531.com | www.616398.com | www.572348.com | www.442095.com | www.026362.com | www.052733.com | www.66731.com | www.7299.club | www.22ta.com | 072898.com | www.740822.cc | www.326708.com | www.339767.com | 219076.com | www.873198.com | www.569536.com | www.388208.com | www.232158.com | www.803089.com | www.523531.com | www.507371.com | www.802820.com | www.20989.com | www.2182.top | www.444089.com | 958901.com | www.947767.com | 637505.com | www.896531.com | www.902613.com | www.841880.com | www.2dj.cc | 奔驰彩票www.509866.com | 591553.com | www.07001.com | www.79ig.com | www.622264.com | www.856108.com | www.908310.com | 大赢家彩票平台www.926730.com | 261802.com | www.275594.com | www.580582.com | www.27nv.com | www.6549.xyz | www.5921.me | www.019642.com | www.11iv.cc | www.dl43.com | www.87407.com | www.2403.vip | www.qq65.com | 大赢家彩票网www.54400x.com | 974848.com | 639476.com | www.x12.com | www.19657.cc | www.sv10.com | 540048.com | 568481.com | www.930213.com | www.794122.com | www.81998.com | www.254959.com | www.33413.com | www.821043.com | www.748482.com | www.608933.com | www.501001.com | www.25955.cc | www.ao13.com | 654924.com | 711162.com | www.699311.com | 662327.com | 050606.com | www.9679.biz | www.m86.mobi | www.481978.com | www.192075.com | www.71776.cc | www.371277.com | www.602037.com | www.511705.com | www.315742.com | www.87890.cc | www.025796.com | www.875905.com | www.117782.com | 762152.com | www.4834.biz | www.63xg.com | 140436.com | www.786023.com | www.944481.com | www.1371.cc | www.4395.cc | www.152960.com | www.077578.com | www.971813.com | 865029.com | 973651.com | www.66932.cc | www.209995.com | www.786255.com | www.750092.com | www.584721.com | www.383763.com | www.973205.com | www.fp40.com | www.dh97.com | 8816彩票www.8816f.com | 388236.com | www.2wq.cc | 99yydb.com | 947607.com | 81079.cc | 554298.com | www.908818.com | www.395919.com | www.57513.com | www.il22.com | www.772010.com | www.892145.com | www.617090.com | www.100074.com | www.730456.com | 294132.com | www.90nk.com | www.33sz.com | www.24100.com | 030918.com | www.66282.cc | www.766534.com | www.736896.com | www.31ed.com | www.7168.org | www.3958.cc | www.17za.com | www.955117.com | 507035.com | www.910058.com | www.270608.com | www.356020.com | 143328.com | www.990604.com | www.661119.com | www.369495.com | www.059116.com | www.8059.wang | www.kn30.com | 221453.com | 201357.com | www.0418.site | www.rt.com | 699223.com | www.417372.com | 453781.com | 809355.com | www.816490.com | www.rp02.com | 500彩票网www.976909.com | 60tk.com | www.297009.com | www.450475.com | www.00151.com | www.63992.cc | www.5mm.com | www.228909.com | www.158358.com | www.884777.com | 082768.com | 18648.com | www.38705.com | 370252.com | www.463013.com | 082014.com | www.936398.com | www.719752.com | www.gp39.com | 151036.com | www.76ir.com | www.823739.com | 248697.com | 859728.com | www.122993.com | www.57708.com | www.607062.com | www.380147.com | www.cx75.com | 3065.biz | www.859412.com | www.468198.com | www.139811.com | www.32934.com | www.08.org | www.cai9466.com | 189061.com | www.ut85.com | www.mg62.com | 615189.com | 366514.com | www.2161.xyz | www.875282.com | www.851862.com | 338042.com | www.833821.com | www.325138.com | www.zn3.com | 930627.com | www.931970.com | 130222a.com | www.833665.com | www.562937.com | www.330653.com | www.038553.com | www.051080.com | www.6503.top | www.gx34.com | 35934.com--35934.com | 866493.com | www.785590.com | www.438634.com | www.176188.com | www.33279.com | www.76ua.com | www.by06.com | 962904.com | 395715.com | www.887636.com | www.690052.com | www.jq44.com | www.160232.com | www.089557.com | www.116392.com | www.441235.com | www.216881.com | www.309432.com | www.179626.com | www.027602.com | www.2738.in | www.593752.com | www.251566.com | www.166961.com | www.31697.cc | www.005780.com | www.020049.com | www.1270.net | www.90pf.com | www.zz65.com | 易彩网www.36166u.com | www.063020.com | www.9949.net | 202815.com | 659047.com | www.827933.com | www.522176.com | www.274040.com | www.914954.com | www.658214.com | www.553038.com | www.365139.cc | www.990184.com | www.758583.com | www.566286.com | www.kx12.com | www.29944.cc | www.163833.com | www.688695.com | 4052.biz | www.864686.com | www.695548.com | www.242477.com | www.0680.vip | www.6ky.com | 373099.com | 52480.cc | www.217048.com | 777144.com | www.cai6661.com | 33415.com | www.8957.xyz | www.957375.com | 994851.com | 901565.com | www.929909.com | www.793174.com | 608716.com | www.151947.com | www.66791.cc | www.326648.com | www.292821.com | www.635189.com | 772422.com | 36644.com | www.127273.com | www.792312.com | www.54211.com | www.1447.cn | www.co25.com | 539244.com | www.717805.com | 8777.hk | www.944107.com | www.344803.com | www.287285.com | www.19733.cc | www.16th.com | www.cai9900.com | www.vl65.com | www.783773.com | www.456615.com | www.742125.com | www.344108.com | www.zd18.com | www.my2.com | www.81162.cc | www.534565.com | www.5fr.com | 715648.com | 628954.com | www.222439.com | www.71332.cc | www.2020.com | www.78er.com | www.wg92.com | 460062.com | 008704.com | 5831.biz | www.755280.com | www.570939.com | www.683192.com | 285355.com | 446876.com | www.693139.com | www.27337.com | www.426692.com | www.673883.cc | 784204.com | www.5zi.cc | www.961101.com | www.600080.cc | www.632476.com | www.301955.com | www.277775.com | www.57351.cc | www.661922.com | www.630609.com | www.4330.vip | www.f17.shop | www.954243.com | www.679156.com | www.380318.com | www.158883.com | www.165696.com | www.877503.com | www.053229.com | www.336753.com | www.092756.com | www.335444.com | www.050130.com | www.092225.com | 500彩票www.338071.com | 712473.com | 62ze.com | 229210.com | 159881.com | www.656668.com | www.817575.com | www.909054.com | 688hz.net | www.751647.com | www.503355.com | www.189142.com | www.886658.com | www.1il.com | www.372037.com | www.126432.com | 801094.com | www.670037.com | www.655395.com | www.590191.com | www.010093.com | www.125441.com | www.493769.com | 中彩网www.71233j.com | www.018668.com | www.5oj.com | www.zr40.com | 448191.com | www.787917.com | www.009475.com | www.806782.com | www.576344.com | www.637162.com | www.376455.com | www.a19.bid | www.41742.com | www.17yu.cc | www.371248.com | 199017.com | www.820746.com | www.301978.com | www.2924.biz | www.37ul.com | www.7568.cc | 杏彩www.00840u.com | 166430.com | www.949054.com | www.563515.com | www.122804.com | www.053797.com | www.193773.com | 爱彩乐www.682918.com | 943368.com | www.48188.com | www.83144.com | www.80jc.com | www.768077.com | www.194007.com | www.068101.com | www.pv7.cc | www.858451.com | 203544.com | www.825172.com | www.906090.com | www.588176.com | www.92aq.com | www.6864.vip | www.5359.cm | 662716.com | 705180.com | 071902.com | 323477.com | 4374.cc | www.986468.com | www.653434.com | www.54677.com | www.dj02.com | 64884.com | www.869786.com | www.883902.com | www.36807.com | www.640132.com | 779597.com | CP701www.627791.com | 491884.com | www.720066.com | www.294619.com | www.0736.me | www.150578.com | www.71qt.com | 555877.com | www.341711.com | www.94168.com | www.309280.com | www.1357.online | www.2727.win | www.193795.com | 490369.com | www.385826.com | www.358373.com | www.338992.com | www.az44.com | www.51697.cc | www.224236.com | www.773412.com | 盛大彩票www.aa0090.com | 656313.com | VIP彩票www.312876.com | www.079097.com | www.571164.com | www.368658.com | www.213022.com | www.101027.com | www.527365.com | www.985046.com | www.942513.com | www.772363.com | www.361312.com | www.94492.com | www.0145.win | www.ek5.com | y85.cc | www.862782.com | www.278796.com | www.8228.vip | www.3ve.com | 433748.com | www.914323.com | www.313670.com | www.454.tv | 665594.com | www.500730.com | 中国福利彩票www.08588z.com | 大赢家彩票www.579031.com | 972848.com | www.669769.com | www.225595.com | www.2326.xyz | www.719437.com | www.958046.com | 468211.com | www.540072.com | www.7638.vip | www.cp0718.com | www.002397.com | www.6015.cn | www.0868.org | www.9us.com | www.59bh.com | www.338643.com | www.6909.xyz | www.65959.com | www.176971.com | www.582161.com | www.xw12.cc | www.873820.com | www.747799.com | www.660755.com | www.205008.com | www.885451.com | www.882714.com | www.976804.com | www.037802.com | www.x80.biz | 612817.com | www.571545.com | 236061.com | www.xz33.com | 360147.com | 811027.com | www.583.date | www.900834.com | www.800195.com | www.323115.com | www.751567.com | www.445067.com | www.1814.live | www.77524.cc | www.mv84.com | 969781.com | www.814399.com | www.4950.cm | www.35sa.com | 233764.com | www.476013.com | www.900182.com | www.440377.com | 959472.com | www.760378.com | www.981116.com | www.582208.com | 500万彩票www.583792.com | 12324.cc | www.599818.com | www.220719.com | www.9728.in | www.qg74.com | www.fk89.com | 562631.com | 026721.com | www.687342.com | www.b30.shop | 127144.com | www.801107.com | www.304169.com | www.916035.com | 337319.com | www.740711.com | 782140.com | 935051.com | www.689598.com | www.248104.com | www.75638.com | www.126638.com | www.42eo.com | 969984.com | cp9818.com | www.851705.com | www.323004.com | www.5253.top | www.gd0.com | 025726.com | www.989876.com | www.208318.com | www.075996.com | www.544590.com | www.098220.com | www.03kj.com | www.204868.com | www.365369.cc | www.251075.com | www.870438.com | www.873630.com | 6012.biz | 560293.com | www.907058.com | www.435886.com | www.879864.com | www.765536.com | www.216819.com | www.26258.com | www.806122.com | www.568949.com | www.127431.com | www.547902.com | www.271024.com | www.576565.com | www.134742.com | www.64222.cc | www.610.cm | www.2011.cm | www.iz7.com | 914422.com | www.877285.com | 47481.com | www.666293.com | www.761145.com | www.43384.com | www.cp99.com | 80811.cc | www.650715.com | www.22886.com | www.990215.com | www.679915.com | www.325395.com | www.20518.com | 520260.com | www.981423.com | www.591380.com | www.165168.com | www.09822.com | 幸运彩票www.550806.com | 442678.com | www.871926.com | www.989948.com | www.556305.com | www.509870.com | www.91398.com | 中彩网www.91233u.com | www.726552.com | www.520577.com | www.947206.com | www.nb22.com | 303923.com | www.475365.com | www.724222.cc | www.293515.com | www.363749.com | www.22308.cc | 吉利彩票www.80065f.com | www.949057.com | www.637073.com | www.533799.com | 中国福利彩票www.6364n.com | www.999233.com | www.445068.com | www.809284.com | www.886583.com | www.266126.com | www.175920.com | www.388372.com | 518674.com | www.33596.cc | 500彩票www.26299s.com | www.i05.cn | 038739.com | www.603768.com | www.070959.com | www.837961.com | 274105.com | 246876.com | www.722489.com | www.074287.com | www.78705.com | www.8825.in | 847481.com | www.979079.com | www.602992.com | www.85698.cc | 572661.com | www.cp931.com | www.878713.com | www.713120.com | www.309749.com | www.9959.pw | www.tv04.com | 995400.com | www.664412.com | 748777.com | www.vl4.com | 997225.com | www.751436.com | www.830139.com | www.654674.com | www.227877.com | www.003425.com | www.521195.com | www.238188.com | www.1888.tv | www.18228.cc | www.70550.com | www.121028.com | www.07xi.com | www.784636.com | www.690615.com | www.933056.com | www.432077.com | www.60438.cc | www.59674.com | www.339428.com | www.658268.com | www.04xn.com | www.ks07.cc | www.7003.vip | www.36vy.com | www.3061.vip | 984739.com | www.889557.com | 七乐彩票www.qilc5.com | 202446.com | www.67jq.com | www.wz03.cc | 210668.cc | www.963429.com | www.099763.com | www.06893.com | 999927.cc | www.206843.com | www.4642.vip | www.111803.com | 2665.wang | www.212189.com | www.61085.com | www.97192.com | www.95kn.com | www.294678.com | www.86138.cc | www.019148.com | www.1518.xyz | www.299347.com | www.921578.com | www.486453.com | www.226938.com | www.3368.com | www.099975.com | www.569708.com | www.886097.com | www.699037.com | www.456791.com | www.5049.org | www.2277.hk | www.5719.com | 716448.com | www.871377.com | www.750532.com | www.256716.com | www.922726.com | www.356276.com | www.700268.com | www.390950.com | www.121770.com | www.081361.com | www.429.com | 福彩www.26878s.com | www.010501.com | www.82719.com | www.ie22.com | www.688843.com | www.110545.com | www.963059.com | 360387.com | www.530333.cc | www.78482.com | www.091087.com | www.812741.com | www.382597.com | www.12431.com | www.868459.com | www.970839.com | www.06577.com | www.978.me | www.5255.cc | www.784788.com | www.220136.com | www.18236.com | www.135081.com | www.3599.cc | 654043.com | 948475.com | www.308203.com | www.97880.com | www.151805.com | www.2332.date | www.cp9775.com | www.650509.com | www.876150.com | www.63qy.com | 909214.com | www.608972.com | www.71747.com | www.296135.com | www.205915.com | www.84qo.com | www.3438.vip | www.999565.com | www.426405.com | www.001908.com | www.1642.vip | 福彩网www.603294.com | 113460.com | www.cai6811.com | www.552326.com | www.249808.com | www.6074.vip | 511592.com | www.821506.com | www.252176.com | www.693.me | 276175.com | www.656697.com | www.211113.com | www.97qn.com | fe70.com | www.542433.com | www.66078.cc | 870514.com--www.wns9778.com | www.566958.com | www.4556.cn | www.989401.com | www.210362.com | www.4853.biz | 566817.com | www.662063.com | www.062029.com | www.hv87.com | www.920098.com | www.299936.com | www.110.com | 055303.com | www.604917.com | www.022113.com | www.cp3766.com | www.880070.com | www.296317.com | www.1312.pw | 001575.com | www.606756.com | www.004328.com | www.tq6.cc | www.888772.com | www.241238.com | www.92os.com | 275342.com | www.588667.com | www.003981.com | 九州彩票www.424988.com | www.830325.com | www.220858.com | www.83of.com | 568188.com | www.555375.com | www.269371.com | www.49xs.com | 336062.com | www.820040.com | www.225820.com | www.64wj.com | 793102.com | www.499429.com | www.24036.cc | 982411.com | www.806727.com | www.155959.com | www.q44.me | www.970854.com | www.327669.com | www.7883.bid | 099568.com | www.759532.com | www.147655.com | www.i74.net | 中彩网www.700345.com | www.984641.com | www.026484.com | www.6703.vip | 730045.com | www.994946.com | www.527959.com | www.93387.cc | www.772402.com | 761834.com | www.778460.com | www.52266.com | 梦想彩票站www.43131i.com | www.bg17.cc | 319086.com | 974787.com | www.0805n.com | www.599032.com | www.467429.com | www.ls89.com | www.31sq.com | 吉利彩票www.66376z.com |