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

彩客吧www.66ckb.com www.978328.com | 458833.com | www.75572.cc | www.785234.com | www.76rh.com | www.594978.com | www.257933.com | www.67fl.com | www.bf49.com | www.93138.cc | 金彩网www.jcai5.com | www.035428.com | www.10tj.com | www.jt47.com | www.912174.com | www.58zm.com | 500彩票www.786587.com | www.69131.cc | www.827772.com | www.84wn.com | www.249343.com | www.1665.wang | www.087371.com | www.152538.com | www.494709.com | 455964.com | www.297339.com | 745522.com | www.kr.cc | www.k56.cc | www.25eq.com | www.345975.com | www.2305.vip | www.254453.com | www.38451.com | www.474880.com | www.83eu.com | www.40846.com | www.596285.com | 天天彩票www.171880.com | www.j38.me | www.085214.com | www.804146.com | www.008081.com | www.249056.com | www.18711.cc | 548699.com | www.9995.loan | www.nl6.com | 947873.com | www.375028.com | www.ie40.com | 529012.com | www.lb89.com | www.4819.vip | www.0071.wang | www.hd17.com | 5763.biz | 150609.com | 871388.com | www.520.me | 44056.com | 422423.com | www.cp4733.cc | www.9081.cm | www.659884.com | www.aa4.com | www.260080.com | 563911.com | www.69490.com | www.789372.com | 093056.com | www.981045.com | www.026324.com | 059871.com | www.612635.com | www.902993.com | www.781522.com | www.555201.com | www.711334.com | www.654003.com | www.231832.com | www.700808.com | 884876.com | 701730.com | www.14wd.com | www.kf08.com | www.cai6123.com | www.464700.com | www.245368.cc | www.923011.com | www.150137.com | www.607328.com | www.949179.com | 112083.com | www.7070.cc | www.020079.com | www.835917.com | 239007.com | www.41pc.com | www.591852.com | www.606881.com | www.985208.com | www.248268.com | www.60355.cc | 629206.com | www.669470.com | www.9959.loan | www.015621.com | www.273815.com | www.44pz.cc | www.088587.com | 113097.com | 009109.com | www.964953.com | www.297772.com | 03ag.com | 727356.com | www.122306.com | www.452013.com | www.754711.com | tk34.com | 065978.com | 184288.com | www.zm74.com | www.384899.com | www.ul43.com | www.238827.com | www.835679.com | www.029928.com | www.781822.com | www.ne01.cc | www.168133.com | www.820021.com | www.802406.com | www.15567.cc | www.115372.com | www.329553.com | www.472889.com | www.155603.com | www.020702.com | www.ml07.com | www.033715.com | www.967814.com | 969416.com | www.7897.top | www.215325.com | www.45xq.com | www.949717.com | www.848536.com | 202489.com | 816392.com | www.vq21.com | www.6080.vip | www.418400.cc | www.940069.com | 238902.com | www.6432.cc | www.408103.com | 456231.com | 456214.com | www.164259.com | 539476.com | www.594066.com | www.258268.cc | www.179762.com | www.010656.com | www.283198.com | www.382093.com | www.526644.com | www.802026.com | www.852667.com | www.074732.com | www.697744.com | www.4956.cc | www.724.website | 295253.com | www.024143.com | www.521309.com | 204524.com | www.90nr.com | www.249055.com | www.562483.com | www.604564.com | 049666.com | www.57550.com | 57dg.com | www.946393.com | 大赢家彩票平台www.183096.com | www.14044.cc | 中国福利彩票www.33588z.com | www.54510.com | www.807884.com | www.509277.com | www.298058.com | 081247.com | www.12128.com | www.66929.cc | www.279053.com | www.663164.com | www.69854.com | www.131187.com | www.665387.com | www.oj42.com | www.172001.com | www.117736.com | www.986590.com | www.37556.com | www.861797.com | www.964871.com | 4075.biz | www.96qa.com | www.3614.cm | www.556358.com | www.699698.com | www.898831.com | www.683128.com | www.50455.com | www.053886.com | www.272351.com | www.82849.com | www.134809.com | www.797622.com | www.938745.com | www.ar78.com | 801991.com | 365471.cc | 234238.com | www.53537.com | www.571892.com | 中国福利彩票www.08588g.com | www.4621.biz | www.125578.com | www.045355.com | www.72931.com | www.54088.com | www.210722.com | 106791.com | 607713.com | www.2456.org | www.384388.com | www.173002.com | 999139.com | www.cr5.com | www.320796.com | www.163907.com | www.518144.com | 网易彩票www.506237.com | 953428.com | 04ko.com | www.38556.cc | www.162569.com | www.ji91.com | 057050.com | www.04206.com | www.751146.com | 688243.com | www.5131.biz | 386375.com | www.737213.com | www.97133.com | www.387339.com | www.3zy.cc | www.023473.com | c73.cc | www.820411.com | www.210352.com | www.8677.date | www.670641.com | www.957374.com | 福利彩票www.8039c.com | www.460051.com | www.820935.com | 天天彩票www.2373f.com | www.8148.cm | 153899.com | www.113234.com | www.635566.com | 978513.com | www.92651.com | www.378036.com | www.2001.cc | www.626171.com | www.oe07.com | www.191211.com | www.512126.com | www.789685.com | 419182.com | 彩乐乐www.cll168.com | www.845649.com | www.hf09.com | www.445333.cc | www.849221.com | www.30lb.com | www.970464.com | www.101801.com | www.668.tv | www.129417.com | www.307358.com | www.052852.com | www.36dj.com | www.391713.com | 150851.com | www.10ds.cc | www.01217.com | www.3446.com | www.228223.com | 058707.com | www.327611.com | www.5586.biz | www.we76.com | www.157298.com | www.949247.com | www.304179.com | www.fd71.com | www.rt03.com | 529637.com | www.12ul.com | www.641887.com | www.nw9.com | www.975272.com | www.840890.com | 623080.com | www.3929.cn | www.205720.com | www.eo87.com | 92779.com | www.5589.site | www.178545.com | www.970845.com | www.9536.online | www.638.cm | www.270902.com | www.382482.com | www.529531.com | www.975748.com | 352808.com | www.ck11.cc | www.715291.com | www.158860.com | www.530419.cc | www.027684.com | www.454566.com | www.663153.com | www.42bs.com | www.109179.com | www.300987.com | www.666032.com | www.4669.vip | www.081220.com | www.277316.com | www.797998.com | www.805365.com | www.019703.com | www.577351.com | 477012.com | 401478.com | www.845196.com | www.121123.com | www.777690.com | www.65op.com | www.8815.loan | 090438.com | www.mx94.com | www.512669.com | www.69504.com | 101066.com | 177186.com | www.cai0866.com | www.85wc.com | www.133553.com | 365427.cc | 931661.com | 374619.com | www.981045.com | 044339.com | www.086368.com | www.446603.com | www.976954.com | 451511.com | www.204026.com | www.871384.com | www.iq10.com | www.xr81.com | www.p45.club | www.26025.com | www.198941.com | www.349837.com | www.540404.com | www.386837.com | www.601853.com | www.742011.com | www.853141.com | www.562790.com | www.nj78.com | www.fw53.com | www.32371.com | www.03700.com | www.075391.com | www.892796.com | www.872104.com | 639010.com | www.32238d.com | 298506.com | www.05335.com | www.412410.com | www.636644.com | www.948049.com | www.14jd.com | www.80669.com | www.916031.com | 656596.com | www.568093.com | www.600469.com | www.500078.cc | www.051586.com | 977335.com | www.009238.com | 501781.com--91838.com | www.666102.com | www.822572.com | 515382.com | www.725424.com | www.277899.com | www.352342.com | 526230.com | 全民福彩www.qmfc8.com | www.195027.com | 716825.com | www.0548.me | www.598820.com | www.54409.com | www.1890.org | www.cp244.com | 017831.com | www.353060.com | www.653271.com | www.678729.com | www.019048.com | www.456470.com | www.672028.com | www.675119.com | www.682270.com | www.jj72.com | www.12tb.com | www.4029.biz | www.438844.com | www.509684.com | 266795.com | 857677.com | 862690.com | www.919541.com | www.312208.com | www.10093.com | 055096.com | www.863587.com | www.365194.cc | www.069823.com | www.399277.cc | 634505.com | www.804017.com | www.65596.cc | www.316018.com | 891337.com | 927116.com | www.1jp.cc | www.36dv.com | www.0236.wang | www.144931.com | www.962096.com | www.533998.com | www.968074.com | www.143666.com | www.381947.com | kv6.com--kv6.com | www.207656.com | www.e55.cc | www.984294.com | www.60964.com | www.024434.com | www.67661.cc | www.oq85.com | ja18.com | 幸运彩票www.982799.com | www.ou55.com | www.956692.com | 957749.com | www.to80.com | 387225.com | www.1398.biz | www.4771.vip | 338241.com | www.48xv.com | www.500911.com | 874385.com | www.s96.club | 6809.me | www.952231.com | www.853075.com | www.938796.com | www.828266.com | 818合彩www.www.1555hc.com | 034935.com | www.53426.cc | www.3665.in | www.xk02.com | www.552331.com | 757509.com | www.87er.com | www.746778.com | 516297.com | www.zr83.com | www.5538.net | www.937310.com | 346056.com | 042133.com | www.5228.pw | www.233167.com | www.723.cm | www.99100.com | www.04709.com | www.86pk.com | 835727.com | www.708192.com | 105924.com | 642629.com | www.1358.cn | www.9344.net | www.889.mobi | www.69673.com | www.713902.com | www.623890.com | www.5544.biz | www.131174.com | www.427543.com | www.744841.com | www.839816.com | www.xg69.com | 706516.com | uedbet.net | www.93355.com | www.130674.com | www.331509.com | www.24811.com | 560257.com | www.315003.com | www.208502.com | www.770044.com | www.75689.com | www.202688.com | www.366840.com | 397868.com | www.683332.com | www.561785.com | www.1087.cn | 548262.com | www.658202.com | www.083610.com | 008099.com | www.139239.com | www.07775.cc | www.730118.com | www.662690.com | www.61282.cc | www.141195.com | www.8088.win | www.r17.cc | www.767336.com | www.222280.com | www.093759.com | www.751150.com | www.632004.com | www.130263.com | www.ai9.cc | www.19866.com | www.819698.com | www.726671.com | www.245314.com | www.xu19.com | 659502.com | www.10lm.com | www.j89.xyz | www.112079.com | www.54044.com | www.175326.com | 974293.com | www.682510.com | www.953909.com | 125505.com | www.027787.com | www.358547.com | www.63kq.com | www.665703.com | www.195312.com | www.593881.com | 161575.com | www.yy71.com | www.1jr.cc | www.279527.com | www.297700.com | 731980.com | www.757200.com | 大赢家彩票平台www.916370.com | 163461.com | www.193876.com | www.882909.com | www.968397.com | www.097810.com | www.921926.com | www.696221.com | www.77152.cc | 818合彩www.www.9889hc.com | www.7381.vip | www.362886.com | www.710715.com | www.984301.com | www.915381.com | www.810402.com | 639894.com | 021909.com | cp1515.com | www.834977.com | www.4535.xyz | www.9259.in | www.07087.com | www.025486.com | www.93865.com | www.384385.com | 31199.cc | www.086983.com | www.ot23.com | www.166082.com | www.95928.com | 543291.com | www.4xs.com | www.872734.com | www.712411.com | www.396344.com | 026789.com | 072410.com | 1cw.cc | www.279731.com | 768090.com | 525936.com | www.cai3655.com | 259723.com | www.596948.com | 015864.com | 039129.com | www.654324.com | www.d52.com | www.441600.cc | 586202.com | 今日彩票www.805462.com | 291040.com | www.091327.com | 福彩www.15355s.com | 920076.com | 067018.com | www.999911.com | www.988535.com | www.663184.com | www.2954.com | www.879228.com | www.sz2.cc | www.122049.com | us28.cc | 532510.com | www.y03.top | 11446.com | www.577085.com | www.4457.net | 305059.com | www.563854.com | www.rj1.com | www.81669.com | www.144384.com | www.95632.cc | www.121000.cc | www.18358.cc | www.304900.com | 127603.com | www.135.website | 979092.com | www.637368.com | www.365812.cc | www.234868.com | www.917969.com | www.810885.com | www.772436.com | www.9059.cc | www.144228.com | www.284566.com | www.572510.com | www.70709.cc | 众乐彩票www.907844.com | www.981874.com | www.163595.com | www.60304.com | www.554.mobi | www.285989.com | www.271698.com | www.909908.com | www.607237.com | www.588745.com | www.392653.com | www.99cp.com | www.775369.com | www.872346.com | www.0015.com | www.8090.com | www.032919.com | www.350439.com | www.004001.com | www.05301.com | 123495.com | www.803545.com | www.823078.com | 057302.com | www.49bt.com | www.04854.com | www.479480.com | www.nm42.com | www.4071.org | www.799216.com | www.622399.com | www.420805.com | www.66sk.com | 243896.com | www.ce30.com | www.404800.com | www.922921.com | www.863640.com | www.106428.com | www.640967.com | 877881.com | www.kj37.cc | www.071882.com | www.05gr.com | www.395406.com | www.gp51.com | www.385193.com | 917603.com | www.h67.in | www.c64.online | www.233007.com | www.a08.top | www.7368.cm | www.853941.com | www.gr62.com | 5000.hk | www.890185.com | www.134593.com | www.js8855b.com | www.903556.com | 972487.com | www.lm55.com | www.339742.com | www.8555.top | www.638196.com | 369647.com | www.246670.com | www.297788.com | www.283.red | www.92339.com | www.171860.com | www.535452.com | 115382.com | 967646.com | www.857159.com | www.534668.com | www.mw43.com | www.at73.com | www.79lu.com | www.93tv.com | www.002277.com | 971483.com | www.43am.com | www.40997.com | www.397277.com | 易彩网www.12455h.com | www.4804.vip | www.189641.com | 852667.com | 280530.com | www.kr08.com | www.6lj.com | 401493.com | 974264.com | www.992131.com | www.654455.com | www.bv78.com | www.la9.com | www.398089.com | www.769937.com | 083953.com | www.132484.com | www.fe37.com | www.731488.com | www.659514.com | www.22801.com | www.531722.com | www.297612.com | 491880.com | www.4907.vip | www.36443.com | www.69815.com | www.480540.com | www.cx30.com | www.51522.cc | www.580153.com | www.629005.com | www.66285.cc | www.695027.com | www.678374.com | www.157122.com | www.013404.com | www.827252.com | www.225160.com | www.609313.com | 445408.com | 23427.com | www.498580.com | www.365800.com | www.621467.com | www.914464.com | 088365.com | 02876.com | www.757392.com | www.885029.com | www.129204.com | www.r20.site | www.278576.com | www.vm62.com | www.785989.com | www.237965.com | 066230.com | www.409961.com | 844324.com | www.789242.com | 2344.bid | 358304.com | 480091.com | 575806.com | www.at71.com | ag52.cc | www.971667.com | www.857186.com | www.se42.com | www.325915.com | www.338789.com | 705858.com | www.875273.com | www.46600.cc | www.619713.com | www.59554.com | 500彩票www.328477.com | www.021192.com | 429321.com | 15674.com--15674.com | www.62bh.com | www.1136.in | www.398.tv | www.5282.net | 872096.com | www.662718.com | www.4574.biz | www.017353.com | www.337109.com | www.895711.com | 4015.biz | 0094.cc | www.49ln.com | www.227161.com | www.329308.com | 631241.com | www.948405.com | 654972.com | www.67pc.com | 560720.com | 637904.com | 588819.com | www.08508.cc | www.30xm.com | 9号彩票www.80767f.com | www.540690.com | www.730666.com | www.796174.com | www.7412.top | www.220295.com | www.826681.com | www.98181.com | www.6767.hk | www.9533.me | 中国福利彩票www.08588u.com | 373333.com | www.cp2043.com | www.br92.com | www.97572.com | www.102790.com | www.596929.com | www.270457.com | www.102124.com | www.006294.com | 375515.com | www.61gc.com | www.630516.com | www.428816.com | 19652.com | 098798.com | www.605118.com | www.24my.com | www.718110.com | www.cp8383.com | www.30sa.com | www.89247.com | www.189001.com | www.858.red | 828805.com | www.057673.com | www.498733.com | www.484805.cc | 003350.com | www.926675.com | www.616793.com | 770492.com | www.at26.com | www.32731.com | ev80.com | www.nd98.com | 365557.cc | 404234.com | www.456970.com | www.990351.com | www.835226.com | www.587101.com | 28494.com | www.264543.com | 8141.biz | www.945896.com | www.02ji.com | 9号彩票www.80767z.com | 456456.com | www.99155.cc | 581418.com | 569171.com | www.656671.com | www.6371.xyz | www.732006.com | www.502037.com | www.317648.com | www.26153.cc | www.367.cm | www.pf05.com | 085780.com | 60113.cc | 072008.com | www.801925.com | www.512053.cc | 549713.com | www.656613.com | 425898.com | www.915639.com | www.598230.com | www.468541.com | www.982816.com | www.469038.com | www.622079.com | 276629.com | www.949241.com | www.19ay.com | www.77265.cc | www.873514.com | www.956279.com | www.s37.cn | www.41757.com | www.506306.com | 948653.com | www.210051.com | www.07076.com | www.36av.cc | www.829877.com | www.331271.com | www.83517.com | www.823868.com | www.678964.com | www.870007.com | www.7254.cc | www.302848.com | www.ps59.com | www.9ce.com | www.397673.com | 632605.com | 550458.com | 358834.com | www.17894.com | www.056682.com | www.hk66.com | 054485.com | 610902.com | www.090999.cc | www.578144.com | 694694.com | www.591902.com | www.188606.com | www.065610.com | www.54120.com | www.384499.com | www.538612.com | www.qk84.com | www.73sp.com | www.52xh.com | www.cx97.com | www.765929.com | www.zg87.com | www.822168.com | 860035.com | www.830318.com | www.947300.com | www.507726.com | www.319646.com | www.219894.com | 097602.com | www.274350.com | www.9775.net | www.8743.com | www.15751.cc | www.820048.com | www.661872.com | 895549.com | www.788739.com | 067652.com | 037628.com | www.uh84.com | www.5048.org | www.zx66.cc | www.384801.com | www.28755.com | www.1156.cm | www.12al.com | 026065.com | 078857.com | www.91tr.com | www.4252.net | www.1064.cm | www.04308.com | www.lb41.com | www.9407.cc | www.cj94.com | 519210.com | 390299.com | www.qn97.com | 网易彩票www.062wy.com | 265665.com | www.017894.com | www.4912.com | www.381447.com | www.143315.com | www.300631.com | www.83578.cc | www.787.in | www.98397.com | www.47409.com | www.6187.top | www.895654.com | www.666399.com | wd7.com | 754037.com | www.po83.com | www.495422.com | 576415.com | www.95li.com | www.86co.com | www.92355.cc | www.139595.com | 804072.com | 彩69www.091181.com | www.vm98.com | www.059377.com | www.vr47.com | www.435528.com | www.709005.com | 4781.biz | www.613996.com | 509602.com | 247311.com | www.658841.com | www.lj34.com | 074896.com | www.nj10.com | www.92mq.com | 福彩www.15355z.com | www.481084.com | 059507.com | www.839252.com | www.8663.live | www.210489.com | www.024302.com | 775914.com | 139518.com | www.4149.net | www.832052.com | 528828.com | www.947671.com | 018923.com | www.857398.com | 辉煌彩票www.2875q.com | www.li25.com | www.770459.com | www.yw42.com | www.97884.com | 190206.com | 657743.com | www.503131.com | 902921.com | www.al70.com | www.302403.com | 博通彩票www.606645.com | 48ja.com | www.390332.com | www.886993.com | www.906173.com | www.620231.com | 957324.com | www.75pb.com | www.jz0.cc | www.14sp.com | www.0078.pw | www.184664.com | www.769292.com | www.4329.in | www.9qi.com | www.6200.cm | www.316.tv | www.026497.com | www.2718.in | www.51681.com | www.993716.com | www.162.cm | www.287018.com | www.7874.vip | www.789312.com | www.723787.com | www.668064.com | www.30mi.com | 477761.com | 159彩票www.159888a.com | www.967740.com | www.07376.com | 057620.com | www.938971.com | www.793183.com | www.543227.com | 025492.com | www.909301.com | www.348.bid | www.37994.com | www.632709.com | www.692229.com | www.56gs.com | www.111187.com | www.042247.com | www.506506.cc | 顶尖彩票www.234265.com | www.8579.loan | www.156123.com | 978964.com | 大赢家彩票平台www.976281.com | www.538951.com | www.101926.com | www.17925.cc | www.uv99.com | www.0617.vip | www.637468.com | 981953.com | www.962916.com | www.623935.com | www.779552.com | 277468.com | www.607758.com | www.11hl.com | www.7kk.cc | www.4644.cc | www.03oj.com | www.u59.com | www.277969.com | www.066504.com | www.33608.com | www.724.cm | www.7268.cm | www.056212.com | www.71081.com | 500彩票www.50788o.com | 949364.com | 018821.com | www.sp9.cc | www.107080.com | www.602651.com | www.716632.com | www.571213.com | www.124173.com | www.558465.com | www.687094.com | 850804.com | 878058.com | 556903.com | www.e38.xyz | www.535.red | www.126471.com | 093003.com | 495031.com | www.592200.com | www.069163.com | www.020748.com | www.6718.com | www.187035.com | www.240247.com | www.52257.com | www.243055.com | www.995484.com | 549721.com | www.930449.com | www.903687.com | www.414955.com | www.874234.com | www.365479.cc | www.356412.com | 80lx.com | www.879136.com | 518427.com | www.20uy.com | 中彩网www.81233b.com | 160278.com | www.021558.com | www.049147.com | www.328978.com | 500彩票www.66653y.com | www.073234.com | www.1592.cn | www.15fp.com | 091322.com | 911141.com | www.786069.com | 454188.com | www.665663.com | 647304.com | www.966634.com | www.1259.vip | www.296606.com | www.704618.com | www.959770.com | www.761592.com | www.555128.com | www.288856.com | www.964711.com | www.tc58.cc | www.879411.com | www.870521.com | www.7pd.com | www.623683.com | www.777552.com | www.441060.com | www.20201.cc | www.492244.com | www.45596.com | www.dr8.cc | www.73315.com | 845113.com | www.xe32.com | www.683056.com | 500彩票www.637477.com | www.978505.com | www.10wb.com | www.ai20.com | 32533.com | 524731.com | 334156.com | www.876119.com | 737127.com | 74993.cc | 899955.com | www.843962.com | www.978800.com | www.531794.com | 820817.com | 皇都彩票www.qj63.com | www.qt93.com | 中彩网www.61233y.com | www.112392.com | www.067236.cc | www.5236.net | www.el01.com | 456024.com | www.948831.com | www.597816.com | www.606556.com | www.94pj.com | www.723639.com | www.703326.com | www.997048.com | 496387.com | www.684171.com | www.370578.com | www.944252.com | www.677345.com | www.313105.com | 502581.com | 083208.com | 167196.com | www.657998.com | www.93ut.com | www.198931.com | www.562868.com | www.54pq.com | www.2151.online | www.371872.com | www.603371.com | www.830234.com | 111057.com | www.392.tv | www.si.com | 716089.com | www.25mx.com | www.629783.com | 875013.com | www.jh03.com | www.82.club | www.7572.win | www.12499.com | www.251912.com | www.47924.cc | www.030085.com | www.016221.com | www.29982.com | www.994881.com | www.9303.vip | www.18854.com | www.559223.com | www.003147.com | www.720579.com | www.915594.com | 979224.com | www.915771.com | www.365479.cc | 571749.com | www.126543.com | 00499.com | 303261.com | www.58631.cc | www.97771.cc | www.011177.com | www.132756.com | 749504.com | www.862923.com | www.193272.com | www.35717.cc | www.791457.com | www.398691.com | www.016596.com | www.89722.cc | www.992176.com | www.603422.com | www.499163.com | www.205656.com | www.17771.com | www.037589.com | www.36225.com | www.015950.com | www.75no.com | l01.net | 967824.com | www.903539.com | www.657545.com | www.69oz.com | www.636334.com | www.390583.com | 371789.com | www.60477.com | 779806.com | 64ar.com | 525553.com | 251385.com | www.932575.com | www.966085.com | www.714014.com | www.850369.com | www.249829.com | www.eg81.com | 386390.com | 790292.com | 810569.com | 金冠彩票www.c4529.com | 397857.com | www.li87.com | 919858.com | www.tz41.com | www.020441.com | www.357022.com | www.675386.com | www.253286.com | www.158310.com | www.669687.com | 893160.com | www.5898.in | www.jj53.com | www.332166.com | 233649.com | 680871.com | hk90.com | www.874462.com | www.5289.com | 天下彩www.249880.com | 946381.com | www.862331.cc | 87rt.com | 500彩票www.652291.com | 954709.com | www.650966.com | www.40tr.com | 840045.com | www.141325.com | www.694801.com | 天下彩www.842686.com | www.681152.com | www.974500.com | www.630083.com | www.695875.com | www.089372.com | www.4344.in | www.139799.com | www.m10.cx | www.697098.com | www.5550.me | 986260.com | www.cp781.cc | 358713.com | www.cp7198.com | 775877.com | 华人彩票www.220755.com | www.720.cx | www.08za.com | 500彩票www.50064g.com | www.396414.com | www.306783.cc | www.272284.com | www.036226.com | www.567448.com | www.215151.com | www.91ju.cc | 8627.pw | www.766884.com | www.0307.in | www.fj43.com | 32842.com | www.613931.com | 761546.com | www.824402.com | www.870484.com | www.970218.com | www.445277.com | www.0536.loan | www.do23.com | 398573.com | www.666254.com | www.221167.com | www.062356.com | www.7pj.com | 164064.cc | www.902520.com | www.cp5685.com | 946541.com | www.741922.com | www.61366.com | www.03401.com | www.35vh.com | www.40tz.com | www.40926.com | www.85619.cc | www.1364.org | www.61287.cc | www.961811.com | 彩多多www.78949q.com | 739120.com | www.72wx.com | www.cp9578.com | 394149.com | 009106.com | www.968302.com | www.700036.cc | 127411.com | 026797.com | www.94wg.com | www.39626.cc | www.8613.top | www.cai7211.com | www.775320.com | www.399351.com | www.769877.com | www.7340.in | www.325517.com | www.4574.top | www.tp02.com | www.522145.com | www.87287.cc | 804923.com | www.c91.top | www.86rs.com | 271883.com | www.175857.com | www.752659.com | www.487164.com | www.222661.com | www.013002.com | www.wa89.com | 985879.com | www.852766.com | www.065683.com | www.868851.com | 大赢家彩票www.697206.com | 664847.com | 328862.com | www.fo52.com | www.865542.com | www.591867.com | www.358459.com | www.134178.com | www.055467.com | www.a22.pw | www.04kh.com | 482965.com | www.961976.com | 522824.com | 647308.com | 599946.com | www.906953.com | www.723958.com | www.862.cm | 5682.cc | www.316647.com | 930417.com--www.361888.com | www.528238.com | 大赢家彩票平台www.163770.com | 664373.com | www.870113.com | 695071.com | www.740818.com | www.505950.com | www.050097.com | www.935.red | www.78vo.com | www.718722.com | www.207557.com | 583752.com | www.692516.com | www.gb18.com | 福彩www.16878n.com | www.913582.com | 083880.com | www.619429.com | www.445231.com | www.977195.com | www.28368.cc | www.3441.org | www.zn58.com | 908448.com | 623071.com | 09yn.com | www.673947.com | www.506328.com | www.062730.com | 960646.com | www.lr80.com | www.68437.cc | www.mn11.com | 984206.com | www.467.tv | www.177263.com | www.123411.cc | www.250.gg | www.e27.net | 965534.com | www.517522.com | www.84pm.com | 844928.com | www.111667.cc | www.817569.com | www.316831.com | www.356000.com | www.220308.com | www.9199.top | 云博彩票www.www.221375.com | www.234328.com | www.106563.com | 618190.com | www.107790.com | www.6266.pw | www.719888.cc | www.789967.com | www.kr05.com | 952547.com | www.915268.com | www.307322.com | www.558848.com | www.28280.com | www.6546.xyz | www.v68.cc | www.aa04.com | 023111.cc | 356448.com | 970204.com | www.876017.com | www.687380.com | www.857969.com | www.8881.cm | www.200653.com | www.163232.com | www.875909.com | www.803913.com | www.500931.com | www.925564.com | www.810547.com | www.739244.com | www.390622.com | www.355902.com | www.853872.com | www.827831.com | www.619720.com | 468504.com | www.504097.com | www.pn91.com | www.576163.com | www.f74.xyz | www.951061.com | www.06311.com | www.929848.com | www.927666.com | www.681980.com | 930795.com | 639034.com | www.978854.com | www.593266.com | www.567413.com | 500彩票www.50064h.com | 365775.com | www.012819.com | www.6909.vip | www.883687.com | www.bb16.com | www.823552.com | www.9a.com | www.961642.com | www.526310.com | www.771441.com | www.6113.shop | www.125129.com | www.194629.com | www.9917.top | www.017891.com | www.613673.com | www.974775.com | www.744422.com | www.033682.com | www.502111.cc | 38770.com--38770.com | www.846508.com | www.959704.com | www.66563.cc | www.873150.com | www.6885.top | www.807339.com | www.48841.com | 180013.com | 029249.com | www.697190.com | www.331064.com | www.496644.com | www.860503.com | www.944138.com | 260869.com | www.286798.com | www.sunbet.2.2.5.com | www.40at.com | 421006.com | www.31dz.com | 99福彩www.768996.com | 863175.com | www.866836.com | 873907.com | www.969296.com | www.456375.com | www.63373.com | www.0656.vip | www.10544.com | www.tc02.com | 0446.com | www.979710.com | www.820908.com | www.331108.com | www.198572.com | www.359996.com | www.505230.com | www.83317.cc | www.45vn.com | 324258.com | www.779182.com | www.424718.com | www.34152.com | www.zj51.com | www.34104.com | www.h11.me | cai932.com | www.734693.com | www.210425.com | 510568.com | 盛彩网www.41518l.com | www.919338.com | www.78187.com | www.7938.wang | www.543614.com | www.339875.com | www.079608.com | 43zq.com | www.735023.com | www.921235.com | www.8588.net | 957410.com | www.356567.com | www.800095.cc | www.ss27.com | www.960646.com | www.293627.com | www.pe84.com | www.799201.com | 3068.biz | 406062.com | www.665749.com | www.177441.com | www.466401.com | www.171012.com | www.760671.com | www.252590.com | www.38128.com | www.252266.com | www.94793.com | www.139474.com | www.059550.com | ew60.com | www.976559.com | www.436193.com | www.910827.com | www.4516.wang | www.802453.com | www.169176.com | www.ct68.com | www.793813.com | www.86uj.com | www.b21.club | www.383636.com | www.218393.com | www.69529.cc | www.53lc.com | www.083965.com | www.486725.com | www.136711.com | www.28ls.com | www.923363.com | www.877882.com | www.1303.me | www.24446.com | www.0sq.com | www.052959.com | www.0880.pw | www.872550.com | www.072728.com | www.we54.com | 339767.com | www.888578.com | 725353.com | 277432.com | www.3654.net | www.08521.com | www.23ez.com | www.74456.com | 888345.com | www.131740.com | 139443.com | www.339661.com | www.dq10.com | 909187.com | www.456304.com | www.tq82.com | www.886180.com | 1545.com | www.518864.com | www.681180.com | www.521430.com | www.630259.com | www.390015.com | www.518970.com | www.029641.com | 038621.com | www.583151.com | www.668944.com | www.303558.com | www.20966.com | www.800785.com | www.441.net | www.rt48.com | www.221580.com | www.4533.vip | 500彩票www.394077.com | www.pg49.com | 900044.com | 398071.com | 542690.cc | www.948113.cc | www.997329.com | www.500774.com | www.086380.com | www.8046.vip | www.oa95.com | www.95zw.cc | www.0895.date | 639754.com | www.993697.com | www.551992.com | www.22209.cc | 964673.com | www.587675.com | www.3374.cm | www.394847.com | 973149.com | www.892711.com | 944955.cc | www.ux05.com | www.771619.com | 647896.com | 484300.com | www.81106.com | www.219741.com | www.452266.com | 186016.com | www.976634.com | 万利www.32123e.com | www.213952.com | www.238867.com | www.04673.com | www.807792.com | www.130461.com | www.287432.com | www.tk81.com | 848550.com | www.95793.com | www.27ng.com | 4379.biz | www.362807.com | www.78002.cc | www.05pw.com | www.566204.com | www.1408.cc | www.518628.com | www.60975.com | www.04577.com | www.374744.com | www.043567.com | www.025832.com | www.33919.cc | 292236.com | www.703128.com | www.260051.com | www.663829.com | www.411163.com | www.849813.com | www.by07.com | 962092.com | www.461114.com | www.8521.biz | www.81zy.com | 217789.com | www.550762.com | www.27728.com | www.00842.com | 316717.cc | www.766504.com | 437122.com | www.030866.com | www.432890.com | www.6474.me | www.09220.com | 彩票5www.199602.com | 877735.com | www.753885.com | www.290538.com | www.jp89.com | www.898032.com | 419413.com | www.258751.com | www.551901.com | www.528759.com | www.505483.com | www.222666.cc | www.068100.com | www.456188.com | www.73382.com | 862200.com | www.925769.com | www.332323.com | 353368.com | www.602332.com | www.501801.com | www.379267.com | www.112218.com | www.re22.com | 981306.com | 508517.com | www.029862.com | www.6vj.com | www.969909.com | www.368896.com | www.425020.com | www.61371.cc | www.191230.com | www.6969.online | www.3168.online | www.011132.com | www.10ud.com | www.sb7.cc | www.114bf.org | www.xc21.com | www.ah89.com | www.779361.com | www.746456.com | www.001674.com | 065285.com | www.947820.com | 幸运彩票www.631799.com | www.796373.com | www.836882.com | www.771138.com | www.510748.com | www.1443.net | 凤凰彩票www.88266z.com | www.997494.com | www.254731.com | www.5gx.com | www.307791.com | www.m94.net | www.ab00.com | www.126622.com | www.876269.com | www.354062.com | www.775221.com | www.635484.com | www.655623.com | www.977289.com | www.380921.com | www.579229.com | www.063256.com | 739429.com | www.642683.com | www.040123.com | 872106.com | www.701242.com | www.35872.com | 157859.com | www.71000.cc | www.92sh.com | www.lu26.com | www.60vb.com | www.sv17.com | www.19pf.com | 312358.com | www.70327.com | 874813.com | www.550643.com | www.xj79.com | www.540752.com | www.61765.com | 055361.com | www.957283.com | www.555356.com | 473384.com | 947303.com | www.519046.com | www.115134.com | www.tq67.com | www.800432.com | www.t87.club | www.077104.com | www.h07.cn | www.422568.com | www.127869.com | www.rv83.com | www.947768.com | www.345057.com | www.011374.com | www.23kn.com | www.47031.com | www.174300.com | www.021740.com | www.57vu.com | 771154.com | www.890982.com | www.277969.com | 百姓彩票www.zuan222.com | www.272800.com | 748834.com | www.035507.com | 49234.com | www.6570.in | 639224.com | www.675819.com | www.494054.com | www.740864.com | www.652902.com | www.380943.com | www.863503.com | 166477.com | 397402.com | www.760698.com | www.213519.com | www.0423.cn | www.81jm.com | www.10ig.com | 869743.com | www.71321.cc | www.025.pink | www.802266.com | ee49.com | www.528690.com | www.156022.com | www.838759.com | www.767192.com | www.448001.com | 购彩网www.917gc.com | www.234940.com | www.668107.com | www.886734.com | www.882080.com | www.536502.com | www.19585.cc | 6532.com | www.756909.com | www.682