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我国临床分子诊断应用的现状及存在的问题

2018-04-01 李金明 检验医学网

我国乙型肝炎病毒(hepatitis B vires,HBV)人群感染率一直较高,上个世纪80年代,在国内不少传染病医院或医院的传染科实验室,采用同位素(32P)标记DNA探针直接膜上斑点杂交的方法检测HBVDNA。但由于检测灵敏度较低(>103拷贝/mL),尽管假阴性结果存在,但假阳性问题较少发生。1989年后,随着PCR技术的引入,从90年代初到中期,全国大到三级甲等医院,小到镇一级的卫生院,


作者:卫生部北京医院,卫生部临床检验中心  李金明

我国临床分子诊断应用及质量管理的回顾

我国乙型肝炎病毒(hepatitis B vires,HBV)人群感染率一直较高,上个世纪80年代,在国内不少传染病医院或医院的传染科实验室,采用同位素(32P)标记DNA探针直接膜上斑点杂交的方法检测HBVDNA。但由于检测灵敏度较低(>103拷贝/mL),尽管假阴性结果存在,但假阳性问题较少发生。1989年后,随着PCR技术的引入,从90年代初到中期,全国大到三级甲等医院,小到镇一级的卫生院,众多的实验室都在开展临床PCR检验,全国生产PCR试剂的厂家也有上百家之多。临床检测应用中出现了大量的假阳性问题,于是,有些临床实验室在检测HBV DNA后通常是先看HBV血清标志物结果,相符即报阳性,不相符报阴性。一个性病痛原体核酸PCR检测,出现阳性结果,则将患者找过来问,有临床症状的,报阳性,否则,报阴性。导致了一临床医生对PCR检验结果的不信任。其影响之大之深,直到现在,在不同的学术场合,乃至一些临床疾病诊疗指南内,一提到PCR,就会说其不可靠,假阳性过高。鉴于上世纪90年代中期临床PCR检验存在的上述问题,卫生部临床检验中心于1998年3月20日在卫生部北京医院召开专家座谈会,与会专家指出了所存在的大量问题,随后不到一个月,卫生部下发了《关于暂停临床基因扩增(PCR)检验的通知》(卫医发[1998]第9号),暂停了PCR在临床检验中的应用。

这种暂停并不意味着PCR技术本身有什么问题,无疑其是一个成熟的技术,问题的存在是应用不规范,一则实验室没有严格的实验室分区设计;二是实验技术人员没有经过临床基因扩增检验相关的质量保证方面知识的培训;三是实验室没有建立有效的质量保证体系;四是当时的试剂方法多为科研用扩增产物初步鉴定所用到的PCR凝胶电泳方法,很不规范。经过充分酝酿和准备,部中心于1997年年底,向全国发出了第一次HBV DNA质评样本进行室问质量评价的预研,1998年正式开展HBV DNA和丙型肝炎病毒(hepatitis C virus,HCV)RNA室间质量评价项目。1999年8月举办了第一期《全国临床基因扩增检验实验室技术人员培训班》。2002年1月卫生部下发了《临床基因扩增检验实验室管理暂行办法》(卫医发[2002]10号),要求凡是开展向患者收费的临床基因扩增检验项目的临床实验室必须按规范进行设置,要有严格的实验室分区,配备必要的仪器设备,人员须接受部临检中心及其指定机构的培训,持证上岗。同时,要建立实验室质量保证体系,编写标准操作程序(standardoperation procedure,SOP)。具备上述硬件和软件后,须通过由部临检中心或省级临床检验中心组织的专家验收,合格后方可正式开展临床基因扩增检验项目。同时,部临检中心下发了《临床基因扩增检验实验室工作规范》(卫检字[2002]9号)。2002年7月深圳市人民医院检验医学部分子生物实验室成为第一家通过验收的临床基因扩增检验实验室。

现状及问题

长期以来,我国的临床分子诊断项目主要为感染性疾病病原体核酸的扩增检测,如HBV、HCV、人类免疫缺陷病毒(human immunodeficiency virus,HIV)、结核分枝杆茵、沙眼衣原体、淋球菌等,采用的方法基本上都是实时荧光PCR,开展的科室主要在医疗机构的检验科,个别在感染科、皮肤科等临床科室的实验室。极少部分为遗传病的产前诊断,如广东、广西和海南地区的地中海贫血的产前基因检测等,采用的方法主要有PCR琼脂糖凝胶电泳和PCR杂交(膜上和芯片)等,开展的科室除了检验科外,相当部分在妇幼保健相关的产前诊断中心的实验室。已接受培训的人员及通过验收的实验室基本上也都是这些实验室。上述实验室相当一部分都经过了5~10年的运行及持续改进培训(各级临床检验中心定期举办的质控和持续改进培训班及会议等),除了有符合分区要求的实验室外,基本上都建立了较完整的实验室质量管理体系,有了较好的质量意识,迅速扭转了临床PCR检验不可靠的局面。

尽管如此,上述实验室仍有需要注意或改进之处,主要在以下几个方面,主要是理念问题。

(1)SOP没有可操作性。这一点是实验室质量管理和标准化的灵魂,如果SOP的编写不是为实际工作,只是为了应付验收检查,则质量管理和标准化就无从谈起。

(2)仪器设备维护和校准不到位。对PCR仪、恒温干浴仪、生物安全桓、测序仪、杂交仪、芯片扫描仪、加样器等仪器设备没有定期的维护,或根本不知道怎么维护。对加样器、扩增仪等的校准由外部机构如仪器厂家、计量部门等进行,没有SOP、没有校准的原始实验记录,并且不清楚仪器设备的技术档案如何建立及其作用。(3)实验记录的形式化及不会归档。

(4)不会分析室内质控结果,有些没有商品质控品的项目不知道质控品从哪来,自然就没有室内质控。

(5)有些项目不参加室间质量评价。

(6)人员的内部培训形式化,不参加相关的质控会议,缺乏持续的外部培训。

此外,也有相当的人员对实验室分区、缓冲间、传递窗和空气流向设计的必要性和正确性的理解不到位。随着新的治疗药物、药物代谢与特定基因多态性关系、药物作用特定基因的突变等的研究进展,以个体化用药治疗为特征的个体化医学正快走走向临床医学的前台,个体化医学得以实施的一个关键性支柱就是以基因及结构变化扣基因表达为检测靶标的个体化分子检测,这些检测除了荧光原位杂交(fluorescence in situ hybridization,FISH)和银增强原位杂交(silver—enhanced in situ hybridization,SISH)检测乳腺癌HER-2/neu基因扩增以及免疫组织化学法(immunohistochemistry,IHC)检测HER2受体蛋白表达状态等少数项目外,基本上都涉及PCR扩增,如基于特异荧光探针的实时荧光PCR、扩增阻滞突变(amplification refractory mutation sys—tem,ARMS)实时荧光PCR、蝎形(scorpion)探针ARMS实时荧光PCR、PCR测序、PCR芯片或膜或微颗粒(如luminex)上杂交等。相关研究的进展,带来了“分子病理学”、“肿瘤基因组学”、“转录组学”和“药物基因组学”等概念。最早接触这些概念者是相关专业和临床科室的一些专家,于是,近年来个体化医学检测在全国的许多医疗机构的病理科、药剂科、肿瘤科、妇产科、内科、甚至胸外科的实验室开展起来,混乱状况与上世纪90年代初中期病原体核酸PCR检测极为相似,其问题主要表现在以下几个方面:

(1)没有规范的实验室分区设计,大多数开展此类检测的实验室目前处于这种状况。

(2)没有任何的实验室质量保证体系,不知道临床基因扩增检验实验室须验收合格后,方能开展临床检验项目。

(3)人员没有经过有资质机构的规范化培训,缺乏质量意识。

(4)开展临床检测,有的使用商品试剂盒,有的是自配试剂但没有严格的试剂配制SOP,有的甚至一部分实验如标本制备和基因扩增在自己实验室完成,测序则像研究生做科研一样,找有关商业测序公司进行。

(5)没有室内质控,或根本不知道如何进行室内质控。

(6)除了极少部分实验室参加了一些国外公司组织实验室间结果比对外,基本上没有定期的室间质量评价或实验室间比对。

做一件事情,没有一个好的过程很难有一个好的结果,至少是出现好的结果的概率极低。临床基因扩增检验亦如此。对临床基因扩增检验中的“过程”(包括分析前、中和后)的有效掌控,最关键的就是靠SOP的按实际S--作详细制订和日常工作中的切实遵循。编写有可操作性SOP的一个基本原则就是,怎么做怎么写,注重最可能影响结果的细节。为使实验室操作者能形象地理解和掌握SOP,可采用文字叙述加照片图示的方法。此外,要指出的是,一个实验室刚建立,由于没有太长的实际检验运行时间,有很多的问题没有想到,因此,编写的SOP可能就会有缺陷,这就需要在以后实际工作中不断根据发生的问题,对文件进行修改,其根本目的是避免同样的问题出现第二次,这也是质量管理的精髓。整个检验过程包括标本采集运送保存、仪器设备使用维护校准、室内质控及其分析、室间质评及其分析衣结果报告与解释等细节,决定于有可操作性的SOP。细节决定成败。

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createdAvatar=https://wx.qlogo.cn/mmopen/mStl88fu4Nd44v3xHnnictn9UpctwtH62EPQ59TMdI9lnqHMgQ16nH7GmGCfpzicsHgjibYVk6vwSpXrC86gMibRkoaIzf5HwSz7/0, createdBy=78cc1691107, createdName=龙胆草, createdTime=Mon Apr 02 08:40:41 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301983, encodeId=02ba30198306, content=了解了解.继续关注, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=93f31631454, createdName=1dd8c52fm63(暂无匿称), createdTime=Mon Apr 02 07:39:04 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301974, encodeId=7a9e3019e470, content=谢谢分享.学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/NUyjXTCJjo6Nx0VGA4QMDzOP8oAgnV4h51EjSfehJ9QxyiaSmw8PB56BhY8KXFSxHCibXrzcy4abrzSibXQPxJ9kzrLcJyhhB8a/0, createdBy=1d3b1672603, createdName=明月清辉, createdTime=Mon Apr 02 07:32:27 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301943, encodeId=b1d030194376, content=学习了很有用不错, beContent=null, objectType=article, channel=null, level=null, likeNumber=97, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/oLAjfB7s1ib3IzoRn621JdKCFUI4XvNELcibocrAVU6aUzEVDSaf3rYH08PUYUiagGqxAEhj8g9WqqNvD1CupIpCrmnR2OrqHZh/0, createdBy=7f181365728, createdName=飛歌, createdTime=Mon Apr 02 06:55:29 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301942, encodeId=176130194273, content=一起学习学习, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-03 yfjms

    学习

    0

  3. 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topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-03 chunjianf

    学习了.谢谢分享

    0

  4. 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topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-03 青山颖钰

    学习了

    0

  5. 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topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-03 jihuaijun1112

    学习学习学习

    0

  6. 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createdAvatar=https://wx.qlogo.cn/mmopen/mStl88fu4Nd44v3xHnnictn9UpctwtH62EPQ59TMdI9lnqHMgQ16nH7GmGCfpzicsHgjibYVk6vwSpXrC86gMibRkoaIzf5HwSz7/0, createdBy=78cc1691107, createdName=龙胆草, createdTime=Mon Apr 02 08:40:41 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301983, encodeId=02ba30198306, content=了解了解.继续关注, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=93f31631454, createdName=1dd8c52fm63(暂无匿称), createdTime=Mon Apr 02 07:39:04 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301974, encodeId=7a9e3019e470, content=谢谢分享.学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/NUyjXTCJjo6Nx0VGA4QMDzOP8oAgnV4h51EjSfehJ9QxyiaSmw8PB56BhY8KXFSxHCibXrzcy4abrzSibXQPxJ9kzrLcJyhhB8a/0, createdBy=1d3b1672603, createdName=明月清辉, createdTime=Mon Apr 02 07:32:27 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301943, encodeId=b1d030194376, content=学习了很有用不错, beContent=null, objectType=article, channel=null, level=null, likeNumber=97, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/oLAjfB7s1ib3IzoRn621JdKCFUI4XvNELcibocrAVU6aUzEVDSaf3rYH08PUYUiagGqxAEhj8g9WqqNvD1CupIpCrmnR2OrqHZh/0, createdBy=7f181365728, createdName=飛歌, createdTime=Mon Apr 02 06:55:29 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301942, encodeId=176130194273, content=一起学习学习, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-02 龙胆草

    学习谢谢分享

    0

  7. 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topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-02 1dd8c52fm63(暂无匿称)

    了解了解.继续关注

    0

  8. 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topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-02 明月清辉

    谢谢分享.学习了

    0

  9. 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2018, time=2018-04-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=302194, encodeId=a3b73021941d, content=学习学习学习, beContent=null, objectType=article, channel=null, level=null, likeNumber=90, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzR8ylhYfiaoAUx2BsaYHVia1IKhG3Cg8CT84rBrn6iaVRkyAibJxBcLEXabOQ5p19vWgDe34ib5ErxNgx/0, createdBy=8a701629084, createdName=jihuaijun1112, createdTime=Tue Apr 03 00:07:15 CST 2018, time=2018-04-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301998, encodeId=2dda30199856, content=学习谢谢分享, beContent=null, objectType=article, channel=null, level=null, likeNumber=99, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/mStl88fu4Nd44v3xHnnictn9UpctwtH62EPQ59TMdI9lnqHMgQ16nH7GmGCfpzicsHgjibYVk6vwSpXrC86gMibRkoaIzf5HwSz7/0, createdBy=78cc1691107, createdName=龙胆草, createdTime=Mon Apr 02 08:40:41 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301983, encodeId=02ba30198306, content=了解了解.继续关注, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=93f31631454, createdName=1dd8c52fm63(暂无匿称), createdTime=Mon Apr 02 07:39:04 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301974, encodeId=7a9e3019e470, content=谢谢分享.学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/NUyjXTCJjo6Nx0VGA4QMDzOP8oAgnV4h51EjSfehJ9QxyiaSmw8PB56BhY8KXFSxHCibXrzcy4abrzSibXQPxJ9kzrLcJyhhB8a/0, createdBy=1d3b1672603, createdName=明月清辉, createdTime=Mon Apr 02 07:32:27 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301943, encodeId=b1d030194376, content=学习了很有用不错, beContent=null, objectType=article, channel=null, level=null, likeNumber=97, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/oLAjfB7s1ib3IzoRn621JdKCFUI4XvNELcibocrAVU6aUzEVDSaf3rYH08PUYUiagGqxAEhj8g9WqqNvD1CupIpCrmnR2OrqHZh/0, createdBy=7f181365728, createdName=飛歌, createdTime=Mon Apr 02 06:55:29 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301942, encodeId=176130194273, content=一起学习学习, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-02 飛歌

    学习了很有用不错

    0

  10. 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2018, time=2018-04-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=302194, encodeId=a3b73021941d, content=学习学习学习, beContent=null, objectType=article, channel=null, level=null, likeNumber=90, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzR8ylhYfiaoAUx2BsaYHVia1IKhG3Cg8CT84rBrn6iaVRkyAibJxBcLEXabOQ5p19vWgDe34ib5ErxNgx/0, createdBy=8a701629084, createdName=jihuaijun1112, createdTime=Tue Apr 03 00:07:15 CST 2018, time=2018-04-03, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301998, encodeId=2dda30199856, content=学习谢谢分享, beContent=null, objectType=article, channel=null, level=null, likeNumber=99, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/mStl88fu4Nd44v3xHnnictn9UpctwtH62EPQ59TMdI9lnqHMgQ16nH7GmGCfpzicsHgjibYVk6vwSpXrC86gMibRkoaIzf5HwSz7/0, createdBy=78cc1691107, createdName=龙胆草, createdTime=Mon Apr 02 08:40:41 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301983, encodeId=02ba30198306, content=了解了解.继续关注, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=93f31631454, createdName=1dd8c52fm63(暂无匿称), createdTime=Mon Apr 02 07:39:04 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301974, encodeId=7a9e3019e470, content=谢谢分享.学习了, beContent=null, objectType=article, channel=null, level=null, likeNumber=55, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/NUyjXTCJjo6Nx0VGA4QMDzOP8oAgnV4h51EjSfehJ9QxyiaSmw8PB56BhY8KXFSxHCibXrzcy4abrzSibXQPxJ9kzrLcJyhhB8a/0, createdBy=1d3b1672603, createdName=明月清辉, createdTime=Mon Apr 02 07:32:27 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301943, encodeId=b1d030194376, content=学习了很有用不错, beContent=null, objectType=article, channel=null, level=null, likeNumber=97, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/oLAjfB7s1ib3IzoRn621JdKCFUI4XvNELcibocrAVU6aUzEVDSaf3rYH08PUYUiagGqxAEhj8g9WqqNvD1CupIpCrmnR2OrqHZh/0, createdBy=7f181365728, createdName=飛歌, createdTime=Mon Apr 02 06:55:29 CST 2018, time=2018-04-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=301942, encodeId=176130194273, content=一起学习学习, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/vi_32/TDiaAWyicHgedzX3wKo3qcxudVJib7AfibfvNeufHRF2jG3Bw8MmZyWjIhtBV1mpJI7Sy9n7RcBKK5t3CuOKuOFxkg/0, createdBy=dd702206933, createdName=wzb521zf, createdTime=Mon Apr 02 06:55:27 CST 2018, time=2018-04-02, status=1, ipAttribution=)]
    2018-04-02 wzb521zf

    一起学习学习

    0

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