Stem Cell Res Ther:使用蛋白质组学和基因表达分析定位马间充质基质细胞表面蛋白质组以鉴定特异性标记物

2018-11-12 MedSci MedSci原创

干细胞具有很大的组织再生潜力,但在干细胞群可用于临床之前,干细胞必须具有一组特异性标记物,这一点至关重要。虽然已经尝试鉴定一组免疫表型标志物以表征马间充质基质细胞(MSCs),但由于在不同实验室中高质量和一致性能的合适抗体的有限性,马MSCs的免疫表型分析仍然具有挑战性。本研究旨在评估通过使用生物素富集和质谱(MS)分析绘制马MSCs表面蛋白质组的策略,并挖掘蛋白质组以寻找属于分化蛋白组簇的潜在马

干细胞具有很大的组织再生潜力,但在干细胞群可用于临床之前,干细胞必须具有一组特异性标记物,这一点至关重要。虽然已经尝试鉴定一组免疫表型标志物以表征马间充质基质细胞(MSCs),但由于在不同实验室中高质量和一致性能的合适抗体的有限性,马MSCs的免疫表型分析仍然具有挑战性。本研究旨在评估通过使用生物素富集和质谱(MS)分析绘制马MSCs表面蛋白质组的策略,并挖掘蛋白质组以寻找属于分化蛋白组簇的潜在马MSCs表面标记。基因表达分析用于验证。

将来源于骨髓(BM)(n = 3)和脂肪组织(AT)(n = 3)的马MSCs扩增至P3,用于(1)细胞分化为中胚层谱系(软骨形成和成骨),(2)通过生物素化富集MSCs表面蛋白,然后通过凝胶内消化分离的蛋白质和nanoLC-MS/MS分析来解析富集的细胞表面蛋白质组,以及(3)RNA分离和定量实时逆转录酶PCR分析CD29、CD44、CD90、CD105、CD166、CD34、CD45和CD79a基因的表达。

结果,通过MS分析富集的MSCs表面蛋白样品,鉴定了1%FDR的总共1239种蛋白质。在这些蛋白质中,在所有六种生物样品中鉴定出939种。鉴定的蛋白质包括指定为分化分类系统簇的19种蛋白质作为潜在的细胞表面靶标。蛋白质和基因表达模式对于常用的阳性MSCs标志物CD29,CD44,CD90,CD105和CD166是阳性的,并且缺乏阴性MSCs标志物CD34,CD45和CD79a。

该研究的结果表明,当没有合适的抗体时,通过生物素化然后通过MS分析富集MSCs表面蛋白质组是表面标志物的免疫表型分型的有价值的替代方案。此外,它们支持基因表达分析作为有价值的对照分析以验证数据。

原始出处:

Louise Bundgaard, Allan Stensballe, et al., Mapping of equine mesenchymal stromal cell surface proteomes for identification of specific markers using proteomics and gene expression analysis: an in vitro cross-sectional study. Stem Cell Res Ther. 2018; 9: 288.

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encryptionId=252ae1803a3, topicName=白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=778d36, createdName=柳叶一刀, createdTime=Sat Jun 01 17:49:00 CST 2019, time=2019-06-01, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1794655, encodeId=66521e946552d, content=<a href='/topic/show?id=82b416e79f5' target=_blank style='color:#2F92EE;'>#stem cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=23, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=16779, encryptionId=82b416e79f5, topicName=stem cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=hb2008ye, createdTime=Fri Apr 05 00:49:00 CST 2019, time=2019-04-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1904518, encodeId=8e2d19045184a, content=<a href='/topic/show?id=b8aa619690e' target=_blank style='color:#2F92EE;'>#标记物#</a>, beContent=null, objectType=article, 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href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Wed May 29 21:49:00 CST 2019, time=2019-05-29, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1754840, encodeId=afcd1e54840fb, content=<a href='/topic/show?id=0a1688e33b3' target=_blank style='color:#2F92EE;'>#蛋白质组#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=20, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88733, encryptionId=0a1688e33b3, topicName=蛋白质组)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=cf6736789983, createdName=jjjiang0211, createdTime=Wed Dec 26 12:49:00 CST 2018, time=2018-12-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1297059, encodeId=1532129e059b5, content=<a href='/topic/show?id=158288e017c' target=_blank style='color:#2F92EE;'>#蛋白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88701, encryptionId=158288e017c, topicName=蛋白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c689267, createdName=cmj8wellington, createdTime=Wed Nov 14 11:49:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
  5. [GetPortalCommentsPageByObjectIdResponse(id=1870984, encodeId=e10318e09847a, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Mon Apr 08 23:49:00 CST 2019, time=2019-04-08, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1936629, encodeId=8a1b1936629b1, content=<a href='/topic/show?id=20f5902e082' target=_blank style='color:#2F92EE;'>#表达分析#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90270, encryptionId=20f5902e082, topicName=表达分析)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=198d2500149, 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encryptionId=252ae1803a3, topicName=白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=778d36, createdName=柳叶一刀, createdTime=Sat Jun 01 17:49:00 CST 2019, time=2019-06-01, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1794655, encodeId=66521e946552d, content=<a href='/topic/show?id=82b416e79f5' target=_blank style='color:#2F92EE;'>#stem cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=23, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=16779, encryptionId=82b416e79f5, topicName=stem cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=hb2008ye, createdTime=Fri Apr 05 00:49:00 CST 2019, time=2019-04-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1904518, encodeId=8e2d19045184a, content=<a href='/topic/show?id=b8aa619690e' target=_blank style='color:#2F92EE;'>#标记物#</a>, beContent=null, objectType=article, 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href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Wed May 29 21:49:00 CST 2019, time=2019-05-29, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1754840, encodeId=afcd1e54840fb, content=<a href='/topic/show?id=0a1688e33b3' target=_blank style='color:#2F92EE;'>#蛋白质组#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=20, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88733, encryptionId=0a1688e33b3, topicName=蛋白质组)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=cf6736789983, createdName=jjjiang0211, createdTime=Wed Dec 26 12:49:00 CST 2018, time=2018-12-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1297059, encodeId=1532129e059b5, content=<a href='/topic/show?id=158288e017c' target=_blank style='color:#2F92EE;'>#蛋白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88701, encryptionId=158288e017c, topicName=蛋白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c689267, createdName=cmj8wellington, createdTime=Wed Nov 14 11:49:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
  6. [GetPortalCommentsPageByObjectIdResponse(id=1870984, encodeId=e10318e09847a, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Mon Apr 08 23:49:00 CST 2019, time=2019-04-08, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1936629, encodeId=8a1b1936629b1, content=<a href='/topic/show?id=20f5902e082' target=_blank style='color:#2F92EE;'>#表达分析#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90270, encryptionId=20f5902e082, topicName=表达分析)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=198d2500149, 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ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1297059, encodeId=1532129e059b5, content=<a href='/topic/show?id=158288e017c' target=_blank style='color:#2F92EE;'>#蛋白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88701, encryptionId=158288e017c, topicName=蛋白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c689267, createdName=cmj8wellington, createdTime=Wed Nov 14 11:49:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
  7. 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href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Wed May 29 21:49:00 CST 2019, time=2019-05-29, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1754840, encodeId=afcd1e54840fb, content=<a href='/topic/show?id=0a1688e33b3' target=_blank style='color:#2F92EE;'>#蛋白质组#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=20, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88733, encryptionId=0a1688e33b3, topicName=蛋白质组)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=cf6736789983, createdName=jjjiang0211, createdTime=Wed Dec 26 12:49:00 CST 2018, time=2018-12-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1297059, encodeId=1532129e059b5, content=<a href='/topic/show?id=158288e017c' target=_blank style='color:#2F92EE;'>#蛋白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88701, encryptionId=158288e017c, topicName=蛋白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c689267, createdName=cmj8wellington, createdTime=Wed Nov 14 11:49:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
  8. [GetPortalCommentsPageByObjectIdResponse(id=1870984, encodeId=e10318e09847a, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Mon Apr 08 23:49:00 CST 2019, time=2019-04-08, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1936629, encodeId=8a1b1936629b1, content=<a href='/topic/show?id=20f5902e082' target=_blank style='color:#2F92EE;'>#表达分析#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90270, encryptionId=20f5902e082, topicName=表达分析)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=198d2500149, createdName=12498bd5m16暂无昵称, createdTime=Fri Apr 26 22:49:00 CST 2019, time=2019-04-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1757139, encodeId=0f8d1e57139db, content=<a href='/topic/show?id=103b421e0e8' target=_blank style='color:#2F92EE;'>#基质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=42170, encryptionId=103b421e0e8, topicName=基质)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=281337296540, createdName=ms7899726347904398, createdTime=Wed Sep 11 00:49:00 CST 2019, time=2019-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1765745, encodeId=48dd1e6574591, content=<a href='/topic/show?id=252ae1803a3' target=_blank style='color:#2F92EE;'>#白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=71803, encryptionId=252ae1803a3, topicName=白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=778d36, createdName=柳叶一刀, createdTime=Sat Jun 01 17:49:00 CST 2019, time=2019-06-01, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1794655, encodeId=66521e946552d, content=<a href='/topic/show?id=82b416e79f5' target=_blank style='color:#2F92EE;'>#stem cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=23, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=16779, encryptionId=82b416e79f5, topicName=stem cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=hb2008ye, createdTime=Fri Apr 05 00:49:00 CST 2019, time=2019-04-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1904518, encodeId=8e2d19045184a, content=<a href='/topic/show?id=b8aa619690e' target=_blank style='color:#2F92EE;'>#标记物#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=22, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=61969, encryptionId=b8aa619690e, topicName=标记物)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=医者仁心, createdTime=Sun Jun 09 03:49:00 CST 2019, time=2019-06-09, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1899253, encodeId=1471189925343, content=<a href='/topic/show?id=6f746e94170' target=_blank style='color:#2F92EE;'>#特异性#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=67941, encryptionId=6f746e94170, topicName=特异性)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=dc7a157, createdName=jinweidong, createdTime=Wed Nov 21 00:49:00 CST 2018, time=2018-11-21, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1961257, encodeId=f5d9196125e73, content=<a href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Wed May 29 21:49:00 CST 2019, time=2019-05-29, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1754840, encodeId=afcd1e54840fb, content=<a href='/topic/show?id=0a1688e33b3' target=_blank style='color:#2F92EE;'>#蛋白质组#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=20, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88733, encryptionId=0a1688e33b3, topicName=蛋白质组)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=cf6736789983, createdName=jjjiang0211, createdTime=Wed Dec 26 12:49:00 CST 2018, time=2018-12-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1297059, encodeId=1532129e059b5, content=<a href='/topic/show?id=158288e017c' target=_blank style='color:#2F92EE;'>#蛋白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88701, encryptionId=158288e017c, topicName=蛋白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c689267, createdName=cmj8wellington, createdTime=Wed Nov 14 11:49:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
    2019-05-29 维他命
  9. 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ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1297059, encodeId=1532129e059b5, content=<a href='/topic/show?id=158288e017c' target=_blank style='color:#2F92EE;'>#蛋白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88701, encryptionId=158288e017c, topicName=蛋白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c689267, createdName=cmj8wellington, createdTime=Wed Nov 14 11:49:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]
  10. [GetPortalCommentsPageByObjectIdResponse(id=1870984, encodeId=e10318e09847a, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Mon Apr 08 23:49:00 CST 2019, time=2019-04-08, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1936629, encodeId=8a1b1936629b1, content=<a href='/topic/show?id=20f5902e082' target=_blank style='color:#2F92EE;'>#表达分析#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=90270, encryptionId=20f5902e082, topicName=表达分析)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=198d2500149, 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href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Wed May 29 21:49:00 CST 2019, time=2019-05-29, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1754840, encodeId=afcd1e54840fb, content=<a href='/topic/show?id=0a1688e33b3' target=_blank style='color:#2F92EE;'>#蛋白质组#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=20, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88733, encryptionId=0a1688e33b3, topicName=蛋白质组)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=cf6736789983, createdName=jjjiang0211, createdTime=Wed Dec 26 12:49:00 CST 2018, time=2018-12-26, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1297059, encodeId=1532129e059b5, content=<a href='/topic/show?id=158288e017c' target=_blank style='color:#2F92EE;'>#蛋白质#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=88701, encryptionId=158288e017c, topicName=蛋白质)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c689267, createdName=cmj8wellington, createdTime=Wed Nov 14 11:49:00 CST 2018, time=2018-11-14, status=1, ipAttribution=)]