这项工作是由部分DK089016和L30 RR030244（MAC），CA153036（AS），P30-DK52574（华盛顿大学消化疾病研究核心）。 AIT是霍华德·休斯医学研究所的医学研究培训研究员。

1。溃疡性结肠炎相关性诱发癌症<ol><li>抛开笼子里的性别和年龄相匹配的6-8周龄小鼠用于实验组和对照组。与尾标记或耳夹的小鼠可以单独标记。 </li><li>在第0天，记录基线的权重，并与10毫克/公斤的保持在-20摄氏度AOM的工作溶液（1 mg / ml的等渗盐水中，10毫克/毫升原液蒸馏水稀释，注入每只小鼠腹膜内（IP）） 。经验的基础上，可以调整剂量7-14 mg / kg和/之间，在实验的早期或重复。 </li></ol> 注意：AOM是一种易挥发的基因毒性剂，并根据附带的MSDS应谨慎处理。稀释液应准备的化学式吸油烟机，保持在冰上，被丢弃后，针对具体机构的协议。 <ol start="3"><li>设为2.5％（2.5 0.5g/100ml的）在蒸馏水中，并通过DSS溶液通过0.22μm的醋酸纤维素fILTER的真空。此剂量可根据小鼠品系和环境的1-3.5％之间进行调整。一旦制备完成，DSS溶液可以保存在冰箱中长达1周。 </li><li>在第7天，供应DSS溶液的小鼠作为他们的饮用水。约250毫升/笼，将需要每次新的DSS提供的最大的5只/笼，但这些都只是估计和动物设施中使用的水瓶的类型将取决于。 </li><li> DSS七天，以提供持续的供应应更换干净的瓶子三次（每2-3天），在此期间，DSS解决方案。一些研究者测量DSS消耗之前更换新的解决方案作为衡量的曝光量。 </li><li>第14天，转笼标准的饮用水了两个星期。 </li><li>重复步骤（1.4） - （1.6）在28和49天的DSS提供了第二个和第三个周期。 “循环”的DSS一个星期的DSS的饮用水其次2周定期（灭菌）水。 </li></ol> 2。临床评估结肠炎与肿瘤进展<ol><li>在小鼠，应权衡利弊/观察，每周2-3次。 ％的重量损失相对基准是用来作为替代措施结肠炎的严重程度。定期评估直肠出血，腹泻，或脱出，可以根据实验目标，并根据不同的评分系统。9，10 </li><li> DSS（每日观察可能是有用的，在这后立即DSS）的一个完整周期后一周的重量损失高达10％，可以预期，随着2-3天的腹泻，直肠出血。 </li></ol>这是可能的，一些老鼠可能无法收回;小鼠失去自己的体重大于20％，是不太容易生存，可能需要提前安乐死。一个单一的IP在这些小鼠注射0.5 - 1.0毫升生理盐水可以成为一个有用的支持措施，对纠正flui<html> D音量因腹泻而失去。 3。小鼠内窥镜检查（可选） <ol><li>内镜的决策支持系统的第二个或第三个周期之后，可以执行之前确认体内肿瘤的生长在牺牲。由于AOM / DSS的模型是相当可靠的，在产生肿瘤，这一步是没有必要的。只有少数几个小鼠应使用这种方法进行评价的，因为它有可能破坏小鼠肿瘤11 </li><li>吸入或注射镇静麻醉的小鼠。 </li><li>小鼠被麻醉后，平面板用胶带固定在鼠标的尾部和胸部。 </li><li>推进小鼠内窥镜插入直肠的鼠标轻轻用生理盐水膨胀，灌溉，改善管腔内查看和删除粪内容。 （请注意，也可以进行内窥镜检查用空气吹入盐水和这种技术作为一种替代，可以提供改善的可视化的肿瘤。12） </li></ol>E“&gt; 4。鼠标的牺牲和结肠收获<ol><li>第70天，每个AOM / DSS对待的鼠标应该怀有多个结肠肿瘤，并准备评估。这个日期可能会延迟几个星期到几个月，如果需要较大的肿瘤。 （值得注意的是，可能需要重度直肠脱垂的小鼠早期被安乐死动物每体制动物护理委员会，以避免过度的不适。） </li><li>尸检之前，单独实施安乐死小鼠异氟醚和颈椎脱位（或其他体制认可的方法）。最终重量和其他测量数据，可在这个时候。 </li><li>奠定其腹侧面暴露砧板上的鼠标。量的腹部，用70％乙醇，以防止毛发污染腹腔内容物，在提取过程中的结肠。 </li><li>使用产钳，把握中线的腹部，做一个小切口，显露腹膜。 </li><li>此切口延伸的腹部的每一侧上沿肋软骨MARGIN。 </li><li>用剪刀剪通过骨盆，使结肠可收获的肛门直肠交界处。这是很重要的，因为DSS结肠炎损伤是最大的在的前端直肠和相应的区域，这是最大的肿瘤的发展。 </li><li>其他组织如肠系膜淋巴结也可以在这个时候收获。 </li></ol> 5。准备结肠宏观分析的<ol><li>使用18G灌胃针连接到一个5毫升注射器，用冰冷的磷酸盐缓冲盐水（PBS），使得流体生理方向退出冲洗结肠。如果结肠组织将被用于RNA或蛋白质为基础的分析，它可以保持在低温，通过采用预先冷却的工作托盘。 </li><li>打开冲洗结肠纵向沿其肠系膜和油漆的冷托盘上。肿瘤是更容易地可视化结肠时，被放置在一个黑暗的表面。可替换地，将1％阿辛兰染料可以施加到突出吨umors。评估肿瘤的数量和大小，用直尺或游标卡尺。 </li><li>切除肿瘤和邻近正常组织的小部分可以在这一点上，供日后分析的RNA，蛋白质或免疫组织化学法（IHC），离开其它结肠组织学评估的某些部分。 </li></ol>数码摄影的总的结肠标本紧随步骤5.2可以帮助肿瘤负荷进行精确的分析，特别是如果冒号的重要部分切除上述其他形式的分析。可以计算出使用自由软件，如ImageJ的肿瘤面积/总结肠区肿瘤的负担（％）。 6。准备结肠组织评估<ol><li>应用10％福尔马林剩余的结肠通过注射器或手指在冰冷的托盘。允许组织固定为至少30秒，以这种方式，有利于转移到固定盆地。确保残留的甲醛是WIP编关闭之前，在同一托盘上画一个新开的结肠。 </li><li>转移冒号充满10％的福尔马林的一个盆地，牵制的边缘。 </li><li>固定3小时后，转移到70％乙醇中的冒号。冒号可以保留无限期地在70％乙醇。 </li><li>三纵截面冒号（近，中，远端/直肠）可以稳定在2％琼脂和石蜡包埋提交。保证在近端结肠和远端结肠/直肠癌取向在一个均匀的方式为每个鼠标。 </li><li>不典型增生，隐窝破坏和炎症可以由有经验的个人描述和评估根据此前公布的协议，10日，13日至15日小鼠可以牺牲前的第70天，如果实验者感兴趣早期发育不良事件的。 </li></ol> 7。代表性的成果 AOM / DSS本文所述的模型使研究人员能够可靠地产生结肠肿瘤在小鼠体内。在这个模型中肿瘤生长的影响直接由相关联的炎症过程。结肠炎严重性应监测临床按照体重减轻和腹泻/便血存在下（ 图2）。这些迹象的疾病活动往往DSS除去后的DSS周期的第5天开始，四个或更多天。在极少数情况下，一个显着的直肠肿瘤负担的小鼠可以发展直肠脱垂。后，第二次或第三次DSS​​周期的腹泻可能成为持续性。通常情况下，肿瘤是本和可辨认的DSS（ 图3）的第三个周期之前由小鼠结肠镜检查。更多的时间是第三个疗程的DSS结果在较大的肿瘤，在收割时（ 图4）。使用的局部应用的爱茜蓝染色可被用来强调肿瘤（ 图5）。结肠肿瘤的照片将协助产生肿瘤测量可用于定量地比较肿瘤生产和大小实验组之间（ 图6）。然后，可以固定，石蜡包埋的结肠标本评价组织学或与使用免疫组织化学染色（ 图7和8）。 <img src="/files/ftp_upload/4100/4100fig1.jpg" alt="图1" /> 图1。图 AOM和DSS管理的。 AOM（10毫克/千克）在第0天被注入。在开始的第二个星期（7天），2.5％DSS溶液给药的小鼠在他们的饮用水。七天的DSS随后两个星期的无菌水。前两个周期的DSS牺牲。 <img src="/files/ftp_upload/4100/4100fig2.jpg" alt="图2" /> 图2。的鼠标重量相对于基线AOM和DSS管理期间，。请注意，在本周每个DSS周期之后，小鼠洛斯<html> Ê5-10％的体重。在本实验中减肥结肠炎严重程度的替代指标。 <img src="/files/ftp_upload/4100/4100fig3.jpg" alt="图3" /> 图3。肿瘤的远端结肠通过小鼠内镜在50天AOM / DSS处理。注意的多个息肉群众阻碍远端结肠的管腔（乙，丙）在比较正常结肠（一）。 <img src="/files/ftp_upload/4100/4100fig4.jpg" alt="图4" /> 图4。纵向打开的小鼠结肠说明肿瘤的大体外观。注意较高的肿瘤负荷在远端结肠/直肠癌（左上图像），和与肿瘤的生长小的近端结肠（右上形象）的特征瓦楞纸纹理。远端结肠，密切了众多肿瘤大小不等（见下文）。 ove_content的“Fo：保持together.within”页面=“总是”&gt; <img src="/files/ftp_upload/4100/4100fig5.jpg" alt="图5" /> 图5。肿瘤所强调的应用阿尔辛蓝染色。注意染料如何强调的结肠的正常纹理以及每个单独的肿瘤的边界。这种染色可以帮助在肿瘤领域由统治者或数字测量的精确测量。 <img src="/files/ftp_upload/4100/4100fig6.jpg" alt="图6" /> 图6。代表分布AOM / DSS每只小鼠的肿瘤治疗的平均次数。注意大多数肿瘤位于远端结肠和&lt;2毫米大小。 <img src="/files/ftp_upload/4100/4100fig7.jpg" alt="图8" /> 图7。石蜡包埋的结肠磁带盒的纵向部分（上述）d的幻灯片后，H＆E染色（如下图）。需要注意的是剩余的阿尔辛蓝染色不干扰H＆E染色。一个大的肿瘤盘旋而上的幻灯片图像（圆圈内）。所用名称“远侧”，“中间”和“近侧”的结果，从切片整个结肠盲肠和肛门之间的三分之二。 <img src="/files/ftp_upload/4100/4100fig8.jpg" alt="图8" /> 图8。代表一个从AOM / DSS管理远端结肠肿瘤的组织学。 H＆E，尿嘧啶和β-catenin的染色切片50X（上图）和400X（底部面板），分别表现出类似人类的结肠腺癌发育不良的变化。

<ol>
	<li>Ekbom, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ulcerative+colitis+and+colorectal+cancer.+A+population-based+study.">Ulcerative colitis and colorectal cancer. A population-based study.</a> N. Engl. J. Med. 323, 1228-1233 (1990).</li><li>Terzic, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammation+and+colon+cancer.">Inflammation and colon cancer.</a> Gastroenterology. 138, 2101-2114 (2010).</li><li>Ullman, T. A., Itzkowitz, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+inflammation+and+cancer.">Intestinal inflammation and cancer.</a> Gastroenterology. 140, 1807-1816 (2011).</li><li>Okayasu, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dysplasia+and+carcinoma+development+in+a+repeated+dextran+sulfate+sodium-induced+colitis+model.">Dysplasia and carcinoma development in a repeated dextran sulfate sodium-induced colitis model.</a> J. Gastroenterol. Hepatol. 17, 1078-1083 (2002).</li><li>Kanneganti, M., Mino-Kenudson, M., Mizoguchi, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+colitis-associated+carcinogenesis.">Animal models of colitis-associated carcinogenesis.</a> J. Biomed. Biotechnol. 342637, (2011).</li><li>Neufert, C., Becker, C., Neurath, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+inducible+mouse+model+of+colon+carcinogenesis+for+the+analysis+of+sporadic+and+inflammation-driven+tumor+progression.">An inducible mouse model of colon carcinogenesis for the analysis of sporadic and inflammation-driven tumor progression.</a> Nat. Protoc. 2, 1998-2004 (2007).</li><li>Tanaka, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+inflammation-related+mouse+colon+carcinogenesis+model+induced+by+azoxymethane+and+dextran+sodium+sulfate.">A novel inflammation-related mouse colon carcinogenesis model induced by azoxymethane and dextran sodium sulfate.</a> Cancer Sci. 94, 965-973 (2003).</li><li>De Robertis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+AOM/DSS+murine+model+for+the+study+of+colon+carcinogenesis:+From+pathways+to+diagnosis+and+therapy+studies.">The AOM/DSS murine model for the study of colon carcinogenesis: From pathways to diagnosis and therapy studies.</a> J. Carcinog. 10, 9 (2011).</li><li>Wirtz, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemically+induced+mouse+models+of+intestinal+inflammation.">Chemically induced mouse models of intestinal inflammation.</a> Nat. Protoc. 2, 541-546 (2007).</li><li>Cooper, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinicopathologic+study+of+dextran+sulfate+sodium+experimental+murine+colitis.">Clinicopathologic study of dextran sulfate sodium experimental murine colitis.</a> Lab Invest. 69, 238-249 (1993).</li><li>Becker, C., Fantini, M. C., Neurath, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+resolution+colonoscopy+in+live+mice.">High resolution colonoscopy in live mice.</a> Nat. Protoc. 1, 2900-2904 (2006).</li><li>Becker, C., Fantini, M. C., Wirtz, S., Nikolaev, A., Kiesslich, R., Lehr, H. A., Galle, P. R., Neurath, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+of+colitis+and+colon+cancer+development+in+mice+using+high+resolution+chromoendoscopy.">In vivo imaging of colitis and colon cancer development in mice using high resolution chromoendoscopy.</a> Gut. 54, 950-954 (2005).</li><li>Shaker, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epimorphin+deletion+protects+mice+from+inflammation-induced+colon+carcinogenesis+and+alters+stem+cell+niche+myofibroblast+secretion.">Epimorphin deletion protects mice from inflammation-induced colon carcinogenesis and alters stem cell niche myofibroblast secretion.</a> J. Clin. Invest. 120, 2081-2093 (2010).</li><li>Boivin, G. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathology+of+mouse+models+of+intestinal+cancer:+consensus+report+and+recommendations.">Pathology of mouse models of intestinal cancer: consensus report and recommendations.</a> Gastroenterology. 124, 762-777 (2003).</li><li>Cooper, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dysplasia+and+cancer+in+the+dextran+sulfate+sodium+mouse+colitis+model.+Relevance+to+colitis-associated+neoplasia+in+the+human:+a+study+of+histopathology,+B-catenin+and+p53+expression+and+the+role+of+inflammation.">Dysplasia and cancer in the dextran sulfate sodium mouse colitis model. Relevance to colitis-associated neoplasia in the human: a study of histopathology, B-catenin and p53 expression and the role of inflammation.</a> Carcinogenesis. 21, 757-768 (2000).</li><li>Yoshida, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+forkhead+box+M1+transcription+factor+contributes+to+the+development+and+growth+of+mouse+colorectal+cancer.">The forkhead box M1 transcription factor contributes to the development and growth of mouse colorectal cancer.</a> Gastroenterology. 132, 1420-1431 (2007).</li><li>Suzuki, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain+differences+in+the+susceptibility+to+azoxymethane+and+dextran+sodium+sulfate-induced+colon+carcinogenesis+in+mice.">Strain differences in the susceptibility to azoxymethane and dextran sodium sulfate-induced colon carcinogenesis in mice.</a> Carcinogenesis. 27, 162-169 (2006).</li><li>Mahler, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+susceptibility+of+inbred+mouse+strains+to+dextran+sulfate+sodium-induced+colitis.">Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis.</a> Am. J. Physiol. 274, 544-551 (1998).</li><li>Nambiar, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preliminary+analysis+of+azoxymethane+induced+colon+tumors+in+inbred+mice+commonly+used+as+transgenic/knockout+progenitors.">Preliminary analysis of azoxymethane induced colon tumors in inbred mice commonly used as transgenic/knockout progenitors.</a> Int. J. Oncol. 22, 145-150 (2003).</li><li>Tanaka, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colorectal+carcinogenesis:+Review+of+human+and+experimental+animal+studies.">Colorectal carcinogenesis: Review of human and experimental animal studies.</a> J Carcinog. 8, (2009).</li><li>Ciorba, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+IDO-1+by+immunostimulatory+DNA+limits+severity+of+experimental+colitis.">Induction of IDO-1 by immunostimulatory DNA limits severity of experimental colitis.</a> J. Immunol. 184, 3907-3916 (2010).</li><li>Kerr, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dextran+sodium+sulfate+inhibition+of+real-time+polymerase+chain+reaction+amplification:+A+poly-A+purification+solution.">Dextran sodium sulfate inhibition of real-time polymerase chain reaction amplification: A poly-A purification solution.</a> Inflamm. Bowel Dis. 18, 344-348 (2012).</li><li>Tang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=is+required+for+resection-induced+changes+in+apoptosis,+proliferation,+and+members+of+the+extrinsic+cell+death+pathways.">is required for resection-induced changes in apoptosis, proliferation, and members of the extrinsic cell death pathways.</a> Gastroenterology. 126, 220-230 (2004).</li></ol>

没有利益冲突的声明。

AOM和DSS的小鼠迅速和有效的模型人结肠炎相关的癌症的治疗。假设关于遗传因素结肠炎相关癌症可以容易地研究与遗传工程小鼠结肠炎相关癌症的效果的药理学目标13，16或者，可以通过采用野生型小鼠研究。 虽然这种模式是高度重视的那些有兴趣在结肠肿瘤的发生发展中的设置炎症的研究存在局限性。在这个模型中小鼠中观察到疾病的程度可以有很大的不同，这取?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> C57BL/6J小鼠</td><td>杰克逊实验室</td><td> 000664 </td><td></td></tr><tr><td>氧化偶氮甲烷（AOM） </td><td> Sigma Aldrich公司</td><td> A5486-100MG </td><td> 储备溶液：稀释至10被保持在-20℃下在蒸馏水中为0.5毫克/毫升- 1毫升的等分试样。 解决方案：1 mg / ml的等渗（0.9％）盐水稀释股票</td></tr><tr><td>葡聚糖硫酸酯钠（DSS） </td><td> TDB咨询</td><td> DB001 </td><td> MW 40 kDa的（36-50 kDa的准备从其他来源是可接受的;应该使用相同的特定的一个单一的实验）6 </td></tr><tr><td> Coloviewminiendoscopic系统</td><td>卡尔斯托斯</td><td>多种</td><td> Becker 等人 11的设备和设置的详细的解释。 </td></tr><tr><td> TPP快速FILTERMAX 500毫升瓶，过滤器，0.22微米PES </td><td>中西部地区科学</td><td> TP99500 </td><td>任何标准组织文化过滤器是可以接受的</td></tr><tr><td>乙醇200证明ASC / USP </td><td>药物AAPER（或其它） </td><td> 11ACS200 </td><td>在蒸馏水中，稀释至70％的</td></tr><tr><td>异氟醚，USP </td><td>巴特勒动物保健供应</td><td> 4029405 </td><td>将鼠标玻璃瓶中，用纱布或小布浸泡在麻醉</td></tr><tr><td> 18G直灌胃针</td><td>布伦特里科学</td><td> N-008 </td><td></td></tr><tr><td>磷酸盐缓冲液（PBS） </td><td>西格玛Aldrich公司</td><td> P5493 </td><td>在蒸馏水中稀释至1X（0.01M） </td></tr><tr><td>冷托盘（II冷板组织康） </td><td> Fisher Scientific则</td><td> NC9491941 </td><td>存放在-20°C </td></tr><tr><td> ImageJ软件</td><td>美国国立卫生研究院（免费下载） </td><td></td><td> <a href="http://rsbweb.nih.gov/ij/" target="_blank">http://rsbweb.nih.gov/ij/</a> </td></tr><tr><td>甲醛（37％） </td><td> Fisher Scientific则</td><td> F79-500 </td><td>在PBS中稀释至10％的</td></tr><tr><td> BD Bacto琼脂</td><td> Fisher Scientific则</td><td> DF0140-01-0 </td><td>使用烤盘创建2％的溶液，在蒸馏水中</td></tr><tr><td> MILTEX眼睛敷料钳</td><td> MedPlus公司</td><td> 18-780 </td><td></td></tr><tr><td> MILTEX眼睛剪刀</td><td> MedPlus公司</td><td> 18-1430 </td> &lt;TD“&gt;”，曲线点期间防止损坏结肠开幕。 </td></tr><tr><td>经Alcian Blue 8GX（粉末） </td><td> Sigma Aldrich公司</td><td> A5268 </td><td> 1克粉末添加到100毫升3％醋酸（3毫升冰醋酸+ 97毫升蒸馏水中） </td></tr><tr><td> 1毫升结核菌素皮锥针注射器附有26克×3/8 </td><td> BD </td><td> 305946 </td><td>对于注射AOM </td></tr></tbody></table>

modeling colitis-associated cancer with azoxymethane (aom) and dextran sulfate sodium (dss)

个人与炎症性肠病（IBD），如克罗恩病（CD）和溃疡性结肠炎（UC）是在健康的个人发展结直肠癌（CRC）的风险增加。这种风险是成正比的持续时间和程度的疾病，结肠发育不良鸡传染性法氏囊病和结肠炎相关癌症（CAC）被认为是发展的结果，累计发病率高达30％，个人长期的UC结肠的广泛参与。上皮细胞的损伤和修复的重复循环，而这些细胞沐浴在一种慢性炎症性细胞因子环境2虽然自发和结肠炎相关的癌症的共享的质量是腺癌，被认为是相关的分子事件的序列是不同的。3这区分认为需要特定动物模型的CAC。 目前存在的一些小鼠模型的研究CAC。硫酸葡聚糖SODIUM（DSS），代理人对结肠上皮细胞的直接毒性作用，可管理的饮用水中创建多个周期的小鼠慢性炎症状态。有足够的时间，这些小鼠肿瘤的发展。4肿瘤的发展，赶紧在这个模型中，如果一个亲致癌设置的管理。这些包括与遗传突变小鼠在肿瘤发生途径（APC，p53基因，MSH2），以及预先处理的基因毒性剂（氧化偶氮甲烷[AOM]，1,2 -二甲基肼[DMH]）。5小鼠与AOM相结合的DSS结肠炎相关癌症的典范，重现性，效力，低廉的价格，和易用性已经得到普及。虽然他们有一个共享的机制，AOM已被发现是更有效和稳定的溶液中比DMH。虽然肿瘤的发生发展中的其他车型，一般需要几个月，注射AOM，随后用DSS小鼠肿瘤的制定适当的的小为7-10周。6，7的最后，AOM和DSS可以施用任何遗传背景的小鼠（基因敲除，转基因等），而不到一个特定的致瘤菌株杂交育种。在这里，我们展示了一个协议驱动的炎症结肠肿瘤在小鼠体内利用一个单一的注射AOM三个7天为一个周期超过10周的时间DSS。这种模式导致肿瘤的组织学和分子的变化密切类似于发生在人类CAC，在本病的发生和化学预防的研究提供了一个非常有价值的模型8

Watch this Scientific Journal Video about Modeling Colitis-Associated Cancer with Azoxymethane AOM and Dextran Sulfate Sodium DSS at JoVE.com

Modeling Colitis-Associated Cancer with Azoxymethane AOM and Dextran Sulfate Sodium DSS

我们展示了一个协议，氧化偶氮甲烷（AOM）的遗传毒性剂，其中给药后由促炎性剂硫酸葡聚糖钠（DSS）的迅速的三个周期的和一贯生成结肠肿瘤在小鼠与在人类结肠炎所见的形态学和分子的相似之处相关的癌症。

建模结肠炎相关的癌症与​​氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）

Individuals with inflammatory bowel disease (IBD), such as Crohn's disease (CD) or ulcerative colitis (UC) are at increased risk of developing colorectal ...

modeling-colitis-associated-cancer-with-azoxymethane-aom-dextran

Research

JoVE Journal

Medicine

Modeling Colitis-Associated Cancer with Azoxymethane (AOM) and Dextran Sulfate Sodium (DSS)

46.5K 次观看.  Washington University School of Medicine. 我们展示了一个协议，氧化偶氮甲烷（AOM）的遗传毒性剂，其中给药后由促炎性剂硫酸葡聚糖钠（DSS）的迅速的三个周期的和一贯生成结肠肿瘤在小鼠与在人类结肠炎所见的形态学和分子的相似之处相关的癌症。

视频: Modeling Colitis-Associated Cancer with Azoxymethane AOM and Dextran Sulfate Sodium DSS

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

建模在小鼠体内的行程 - 中东与灯丝模型大脑中动脉闭塞

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

科研

医学

Modeling and Imaging 3-Dimensional Collective Cell Invasion

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy tissue is the root cause of disease and death. For other cancers (e.g. breast, lung, etc.), it is the process of metastasis, in which tumor cells move from a primary tumor mass, colonize distal sites and ultimately contribute to organ failure, that eventually leads to morbidity and mortality 3. It has been estimated that invasion and metastasis are responsible for 90&#37; of cancer deaths 4. As a result, there has been intense interest in identifying the molecular processes and critical protein mediators of invasion and metastasis for the purposes of improving diagnosis and treatment 5.
A challenge for cancer scientists is to develop invasion assays that sufficiently resemble the in vivo situation to enable accurate disease modeling 6. Two-dimensional cell motility assays are only informative about one aspect of invasion and do not take into account extracellular matrix (ECM) protein remodeling which is also a critical element. Recently, research has refined our understanding of tumor cell invasion and revealed that individual cells may move by elongated or rounded modes 7. In addition, there has been greater appreciation of the contribution of collective invasion, in which cells invade in strands, sheets and clusters, particularly in highly differentiated tumors that maintain epithelial characteristics, to the spread of cancer 8.
We present a refined method 9 for examining the contributions of candidate proteins to collective invasion 10. In particular, by engineering separate pools of cells to express different fluorescent proteins, it is possible to molecularly dissect the activities and proteins required in leading cells versus those required in following cells. The use of RNAi provides the molecular tool to experimentally disassemble the processes involved in individual cell invasion as well as in different positions of collective invasion. In this procedure, mixtures of fluorescently-labeled cells are plated on the bottom of a Transwell insert previously filled with Matrigel ECM protein, then allowed to invade "upwards" through the filter and into the Matrigel. Reconstruction of z-series image stacks, obtained by confocal imaging, into three-dimensional representations allows for visualization of collectively invading strands and analysis of the representation of fluorescently-labeled cells in leading versus following positions.

Models of tumor cell invasion into three-dimensional extracellular matrix better reflect the in vivo situation than two-dimensional motility assays. Using matrix invasion assays combined with confocal imaging of fluorescently-labeled cells, detailed information on invasion modes and the distinct contributions of leading versus following cells can be obtained.

3维建模和成像集体细胞的侵袭

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy ...

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical treatments and improved platinum-based chemotherapies, there has been little improvement in EOC survival rates for more than four decades 1,2. Whilst stage I tumors have 5-year survival rates &gt;85%, survival rates for stage III/IV disease are &lt;40%. Thus, the high rates of mortality for EOC could be significantly decreased if tumors were detected at earlier, more treatable, stages 3-5. At present, the molecular genetic and biological basis of early stage disease development is poorly understood. More specifically, little is known about the role of the microenvironment during tumor initiation; but known risk factors for EOCs (e.g. age and parity) suggest that the microenvironment plays a key role in the early genesis of EOCs. We therefore developed three-dimensional heterotypic models of both the normal ovary and of early stage ovarian cancers. For the normal ovary, we co-cultured normal ovarian surface epithelial (IOSE) and normal stromal fibroblast (INOF)&nbsp;cells, immortalized by retrovrial transduction of the catalytic subunit of human telomerase holoenzyme (hTERT) to extend the lifespan of these cells in culture. To model the earliest stages of ovarian epithelial cell transformation, overexpression of the CMYC oncogene in IOSE cells, again co-cultured with INOF cells. These heterotypic models were used to investigate the effects of aging and senescence on the transformation and invasion of epithelial cells. Here we describe the methodological steps in development of these three-dimensional model; these methodologies aren't specific to the development of normal ovary and ovarian cancer tissues, and could be used to study other tissue types where stromal and epithelial cell interactions are a fundamental aspect of the tissue maintenance and disease development.

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

We describe methodologies for establishing in vitro heterotypic three-dimensional models comprising ovarian fibroblasts and normal ovarian surface or ovarian cancer epithelial cells. We discuss the use of these models to study stromal-epithelial interactions that occur during ovarian cancer development.

异型三维<em在体外</em&gt;卵巢癌的发生和发展过程中的间质 - 上皮细胞相互作用模型

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical ...

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

视网膜脱离模型鼠害由透明质酸钠的视网膜下注射

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully recapitulate early- and late-stage metastatic disease progression. Typical tumor implantation models utilize ectopic or orthotopic primary tumor implantation, but few include systemic spontaneous metastatic disease that mimics the clinical setting. This protocol describes the key steps to develop RCC disease progression stages similar to patients. First, it uses a highly metastatic mouse tumor cell line in a syngeneic model to show orthotopic tumor cell implantation. Methods include superficial and internal implantation into the sub-capsular space with cells combined with matrigel to prevent leakage and early spread. Next it describes the procedures for excision of tumor-bearing kidney (nephrectomy), with critical pre- and post- surgical mouse care. Finally, it outlines the steps necessary to monitor and assess micro-and macro-metastatic disease progression, including bioluminescent imaging as well provides a detailed visual necropsy guide to score systemic disease distribution. The goal of this protocol description is to facilitate the widespread use of clinically relevant metastatic RCC models to improve the predictive value of future therapeutic testing.&#160;

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

Models of spontaneous metastatic renal cell carcinoma (RCC) disease progression can be used for evaluating treatments in a clinically relevant setting. This protocol demonstrates different procedures for orthotopic kidney tumor cell implantation, proper nephrectomy, and finally outlines a necropsy guide for visual and bioluminescent scoring of metastatic burden and localization.

在小鼠模型自发转移性肾细胞癌（MRCC）肾切除

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

Working with Human Tissues for Translational Cancer Research

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human specimens, precious samples must be handled with the greatest care in order to ensure integrity of organs, tissues, and cells to the highest degree. Unfortunately, tissue processing by definition requires extraction of tissues from the host, a change which can cause great cellular stress and have major repercussions on subsequent analyses. These stresses could result in the specimen being no longer representative of the site from which it was retrieved. Therefore, a strict protocol must be adhered to while processing these specimens to ensure representativeness. The desired assay(s) must also be taken into consideration in order to ensure that an optimal technique is used for sample processing. Outlined here is a protocol for tissue retrieval, processing and various analyses which may be performed on processed tissue in order to maximize downstream production from limited tissue samples.

Translational cancer research is dependent on extraction of human tissues. Much work has gone into optimizing extraction methods for ex vivo analysis. Here, we describe tissue processing methods allowing for maximal data output from limited samples.

与人体组织转化为癌症研究工作

Medical research for human benefit is greatly impeded by the necessity for human tissues and subjects. However, upon obtaining consent for human ...

Modeling Encephalopathy of Prematurity Using Prenatal Hypoxia-ischemia with Intra-amniotic Lipopolysaccharide in Rats

Encephalopathy of prematurity (EoP) is a term that encompasses the central nervous system (CNS) abnormalities associated with preterm birth. To best advance translational objectives and uncover new therapeutic strategies for brain injury associated with preterm birth, preclinical models of EoP must include similar mechanisms of prenatal global injury observed in humans and involve multiple components of the maternal-placental-fetal system. Ideally, models should produce a similar spectrum of functional deficits in the mature animal and recapitulate multiple aspects of the pathophysiology. To mimic human systemic placental perfusion defects, placental underperfusion and/or chorioamnionitis associated with pathogen-induced inflammation in early preterm birth, we developed a model of prenatal transient systemic hypoxia-ischemia (TSHI) combined with intra-amniotic lipopolysaccharide (LPS). In pregnant Sprague Dawley rats, TSHI via uterine artery occlusion on embryonic day 18 (E18) induces a graded placental underperfusion defect associated with increasing CNS damage in the fetus. When combined with intra-amniotic LPS injections, placental inflammation is increased and CNS damage is compounded with associated white matter, gait and imaging abnormalities. Prenatal TSHI and TSHI+LPS prenatal insults meet several of the criteria of an EoP model including recapitulating the intrauterine insult, causing loss of neurons, oligodendrocytes and axons, loss of subplate, and functional deficits in adult animals that mimic those observed in children born extremely preterm. Moreover, this model allows for the dissection of inflammation induced by divergent injury types.

Encephalopathy of prematurity encompasses the central nervous system abnormalities associated with injury from preterm birth. This report describes a clinically relevant rat model of in utero transient systemic hypoxia-ischemia and intra-amniotic lipopolysaccharide administration (LPS) that mimics chorioamnionitis, and the related impact of infectious stimuli and placental underperfusion on CNS development.

早产儿脑病建模使用产前缺氧缺血大鼠羊膜腔内毒素

Encephalopathy of prematurity (EoP) is a term that encompasses the central nervous system (CNS) abnormalities associated with preterm birth. To best ...

Modeling the Early Steps of Ovarian Cancer Dissemination in an Organotypic Culture of the Human Peritoneal Cavity

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. Instead, ovarian cancer cells exfoliated from the primary tumor are carried by peritoneal fluid to metastatic sites within the peritoneal cavity. These sites, most notably the abdominal peritoneum and omentum, are organs covered by a mesothelium-lined surface. To investigate the processes of ovarian cancer dissemination, we assembled a complex three-dimensional culture system that reconstructs the lining of the peritoneal cavity in vitro. Primary human fibroblasts and mesothelial cells were isolated from human omentum. The fibroblasts were then mixed with extracellular matrix and covered with a layer of the primary human mesothelial cells to mimic the peritoneal and omental surfaces encountered by metastasizing ovarian cancer cells. The resulting organotypic model is, as shown, used to examine the early steps of ovarian cancer dissemination, including cancer cell adhesion, invasion, and proliferation. This model has been used in a number of studies to investigate the role of the microenvironment (cellular and acellular) in early ovarian cancer dissemination. It has also been successfully adapted to high throughput screening and used to identify and test inhibitors of ovarian cancer metastasis.

Here, we present a protocol to construct a three-dimensional in vitro model of the lining of the peritoneal cavity, composed of primary human mesothelial cells and fibroblasts layered with extracellular matrix, as a tool to investigate ovarian cancer cell adhesion, invasion, and proliferation.

造型卵巢癌传播在人类腹腔的器官文化的早期步骤

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. ...

Modeling Chemotherapy Resistant Leukemia In Vitro

It is well established that the bone marrow microenvironment provides a unique site of sanctuary for hematopoietic diseases that both initiate and progress in this site. The model presented in the current report utilizes human primary bone marrow stromal cells and osteoblasts as two representative cell types from the marrow niche that influence tumor cell phenotype. The in vitro co-culture conditions described for human leukemic cells with these primary niche components support the generation of a chemoresistant subpopulation of tumor cells that can be efficiently recovered from culture for analysis by diverse techniques. A strict feeding schedule to prevent nutrient fluxes followed by gel type 10 cross-linked dextran (G10) particles recovery of the population of tumor cells that have migrated beneath the adherent bone marrow stromal cells (BMSC) or osteoblasts (OB) generating a "phase dim" (PD) population of tumor cells, provides a consistent source of purified therapy resistant leukemic cells. This clinically relevant population of tumor cells can be evaluated by standard methods to investigate apoptotic, metabolic, and cell cycle regulatory pathways as well as providing a more rigorous target in which to test novel therapeutic strategies prior to pre-clinical investigations targeted at minimal residual disease.

Modeling Chemotherapy Resistant Leukemia In Vitro

The current report summarizes a protocol that can be utilized to model the influence of the bone marrow microenvironment niche on leukemic cells with emphasis placed on enrichment of the most chemoresistant subpopulation.

建模化疗耐药白血病<em&gt;体外</em

It is well established that the bone marrow microenvironment provides a unique site of sanctuary for hematopoietic diseases that both initiate and ...

Modeling and Simulations of Olfactory Drug Delivery with Passive and Active Controls of Nasally Inhaled Pharmaceutical Aerosols

There are many advantages of direct nose-to-brain drug delivery in the treatment of neurological disorders. However, its application is limited by the extremely low delivery efficiency (&#60; 1%) to the olfactory mucosa that directly connects the brain. It is crucial to develop novel techniques to deliver neurological medications more effectively to the olfactory region. The objective of this study is to develop a numerical platform to simulate and improve intranasal olfactory drug delivery. A coupled image-CFD method was presented that synthetized the image-based model development, quality meshing, fluid simulation, and magnetic particle tracking. With this method, performances of three intranasal delivery protocols were numerically assessed and compared. Influences of breathing maneuvers, magnet layout, magnetic field strength, drug release position, and particle size on the olfactory dosage were also numerically studied.
From the simulations, we found that clinically significant olfactory dosage (up to 45%) were feasible using the combination of magnet layout and selective drug release. A 64 -fold higher delivery of dosage was predicted in the case with magnetophoretic guidance compared to the case without it. However, precise guidance of nasally inhaled aerosols to the olfactory region remains challenging due to the unstable nature of magnetophoresis, as well as the high sensitivity of olfactory dosage to patient-, device-, and particle-related factors.

This manuscript reviews the modeling and simulations of different protocols to deliver medications to the olfactory region in image-based nasal airway models. Multiple software modules are used to develop the anatomically accurate nose model, generate computational mesh, simulate nasal airflows, and predict particle deposition at the olfactory region.

建模与嗅觉药物传送的模拟与鼻吸入气雾剂药品的被动和主动控制

There are many advantages of direct nose-to-brain drug delivery in the treatment of neurological disorders. However, its application is limited by the ...

建模结肠炎相关的癌症与​​氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）

建模结肠炎相关的癌症与氧化偶氮甲烷（AOM）和葡聚糖硫酸钠（DSS）