这项研究获得资助联邦教育部（BMBF）通过授予公务员事务局（0801）01 EO。柏林中风研究中心（CSB）是一家综合性的研究和治疗中心。我们感谢基金会费尔根豪尔的财政支持。 我们感谢理查德·A·达奇的英文文本更正。

1。制备研究职位<ol><li>让病人采取一个座位，在屏幕的正前方的距离为2 m，（203×152厘米覆盖在横轴上的可视角度的58.15度和43.61度的视觉上的垂直轴的角度，分辨率:1400× 1050），在模拟真实的汽车座椅固定基地模拟汽车座椅。帮助病人调整座椅的距离与手柄下方的踏板。帮助调节靠背。 </li><li>指导患者如何使用仿真车小工具（刹车，转向灯，方向盘）。 </li><li>指导病人任务:在驱动器中，你会做一个真正的非模拟驾驶的情况。这条路是单向车道公路的曲线（最小半径500米，最大半径1200米）和交通。要提高警惕，路牌和突破的道路两侧新兴的汽车。阵营的概念有潜在危险的活动，如野猪或接近的球通过踩下制动或使用转向信号，或两者兼而有之，似乎是适当的在各自的驱动情况下尽快路。同时按下踏板时，汽车加速70公里每小时的恒定速度，除非制动器被使用1。该驱动器大约需要10分钟。 </li><li>通知关于模拟疾病的病人。如果出现不适，恶心，出汗，中断测试环节。 </li><li>用更少的任务密度习惯的模拟情况进行的测试驱动器和允许足够的时间来调整，以防止模拟疾病的方法。 </li></ol> 2。眼动仪的校准<ol><li>在第二次测试环节，术后病人正确就位，并已经获得了足够的练习时间，将眼跟踪病人的头部和拉动灵活伸缩调整它。头摄像头激光指向屏幕中间。调整打印头相机的聚焦的瞳孔。 </li><li>指导患者根据校准鼠标箭头的领导五个点看。 </li><li>开始模拟。 </li><li>完整的校准与添加水平校准:病人注视叠加在左边的屏幕上的图象（眼睛），接着在屏幕上移动的眼睛，并再次注视它的右侧。 </li><li>测试的校准通过询问患者的注意力集中在屏幕上的特定对象上，和匹配用的重叠眼的图片，这表示由软件计算出的视线位置。校准是成功的，如果满足的目光和叠加图像在屏幕上的相同点。眼动仪视力的垂直漂移的过程中可能出现的驱动器。评价通过目视检查在开始和结束时的驱动器的漂移量，检查需要复检。 </li><li>如果校准成功，关闭覆盖图片。如果没有重复校准过程，直到成功为止。 </李&gt; <li>对于快速评估补偿行为的目光运动反过来覆盖眼睛的图片。 </li></ol> 3。模拟<ol><li>通过询问患者开始驾驶进行模拟。 </li><li>由于周围环境的分心水平与不同任务难度，让患者的驱动器不同的路由（每6,500米的农村地区，持续时间约10分钟）。每个病人驱动三条路线。 </li><li>即时评估的动眼神经行为:开启左眼图像上叠加和可视化的注视行为的病人在测试过程中会:眼跟踪连续发送坐标的实际视线位置的仿真软件SILAB的。作为回报，SILAB项目的覆盖眼睛的图片，这是一个形象的眼睛，在屏幕上正好在现场的病人看起来。这不仅可以被用来证明质量校准，也使的目光立即可见的行为不仅给你，但一个莱索托的病人。 </li></ol> 4。分析<ol><li>用于数据记录的使用SILAB软件为100 Hz的采样速率。使用SILAB软件，记录速度，反应时间（使用转向灯，制动）。 </li><li>头部和眼睛运动参数进行统计分析，用Matlab（MathWorks公司，Natic​​k郡，USA）。使用以下条件: <ol><li>作为视线轨迹凝视速度超过30℃/ s和凝视振幅是大于1°（如小于1°的眼球运动属于到microsaccades）的章节定义扫视。集群扫视发生在80 ms内。定义部分之间扫视作为注视。定义头部运动运动超过6°/秒11，幅度超过3°。且不同时头部和眼睛的运动，与目录中的相反的方向，因为它们代表没有凝视振幅增益。 </li><li>作为固定的对象注视的对象上定义视线位置最大1，24＆D例如，除了从物体在x-轴和1个，在y-轴的66°。对象不触发根据患者注视位置，但考虑对象的偏心注视着位置，通过计算它的对象时，会出现3。 </li><li>计算参与者的录制品（平均固定的持续时间），并在水平和垂直的经络（固定位置的方差）的蔓延的搜索的平均长度。 </li></ol></li><li>由任一固定或手动检测的第一检测的方式有两种:作为一个第一模式（第一检测）测量反应时间的测量反应时间:如果病人注视对象的第一个，和响应手动之后（在大多数情况下），然后选择作为第一次检测的反应时间的固定时间。如果病人使用转向灯或制动踏板的一个指标，而不录对象，然后选择手动反应时间为首次发现。作为第二模式（手动反应），测量反应离子手动反应（制动或转信号）。 </li></ol> 5。代表性的成果我们招募了6例不完全偏盲缺血性PCA梗死的权利和2在左侧半球（4）和85名健康对照者的不同年龄段（20-75岁不同年龄段（35-71岁），同样分布），以确定年龄相关的变化在眼睛和头部的运动驾驶性能以及为参照组。他们没有认知功能障碍，神经或精神缺陷或疾病报告和视力高于0.5。病史和虚拟媒体的经验与探索。这项研究是符合赫尔辛基宣言，是当地的伦理委员会批准。从所有参与者获得书面知情同意书。所有受试者不知道的实验的目的。 在这里，我们展示了重新表象的两个病人的测试结果测试7 - 9个月后的右侧和中风事件不完全偏盲（ 图1）无补偿行为以及健康的受试者作为控制。由于年龄相仿，驾驶和电脑游戏的经验，选择健康对照。 患者A代偿扫视运动的视觉缺陷导致正常的表现，在驾驶模拟成功的检测和反应在乡村驾驶的情况下可能产生的危害健康对照相比，位于侧面。然而，乙病人没有表现出代偿扫视运动，并透露表现不佳，驾驶模拟错过了周围物体的盲区，造成长期的反应时间或碰撞。然而，在整个驱动器，乙病人通过代偿行为造成更少的碰撞，没有被指示这样做。测试进行头奔放的，让现实条件和检测头部运动补偿行为可能造成的影响。 患者被要求驱动的过程，因为他们会做一个真正的非模拟驾驶的情况。的健康受试者病人相比甲进行扫视的1.7倍，更频繁，这主要覆盖其中视觉缺陷位于屏幕侧（63％）。扫视病人的振幅A和控制是相似的（平均振幅:5.5度和5.3度的病人的健康问题）。病人的固定时间短健康对照（患者A与控制在483毫秒的平均注视时间为381毫秒）。 对比度患者B和控制探索同样频繁两侧的屏幕上。 图2示出在屏幕上的分布的录制品期间，第一驱动器的patient A，健康者和病人B.的患者B进行3.4少扫视运动相比，患者的覆盖幅度的一半大小的病人（平均幅度:5.5患者A在乙病人为2.9度）。患者B表现出较长的固定持续时间比较健康的控制和患者的A（平均注视时间1049毫秒）。 病人和病人乙的表现几乎没有头运动（1〜2），而健康对照执行少数（5〜10）头架次驱动会话促进凝视幅度。 图3显示的对象的位置的偏心的影响，相对于视线位置上的反应时间分别为左侧和右侧的视场，证明。该图中示出了反应时间的增加，由于偏心所示的视场的两侧分开在两个主题。在非常小的ECC一些反应时间<html> entricities是小于50 ms。这些都不是真实的反应时间，但而由于扫描可能的危险场所，公路沿线，或点固定的患者在出现的对象。我们没有过滤这些事件，因为这也代表了一定的兴趣:承认驾驶行为产生潜在的危险场所。 （该图还显示，有少的反应时间，注意病人的B，因为在他的盲区错过的对象。） 在病人和健康对照的所有对象进行检测，并没有冲突发生。虽然在乙病人，不同的反应时间清楚（盲人）之间的权利和左（近视）中:患者B检测对象相比，明眼人领域中发生的盲区慢了1.6倍和4倍相撞的对象发生在盲区（反应时间中位数:右（盲人）领域:4411.66毫秒与左（近视）领域的2810毫秒）。 "因此，病人的补偿他的损失以及由右偏心视觉扫视运动，达到侧视野缺损的人数有所增加。目前还不清楚，但如果这种代偿策略变得具有较高的工作量不足。证据是建议虽然病人管理同样快的反应的右侧由于偏侧扫视运动，在图中为左视野:他表明较长的反应时间，在更大的偏心率的左侧，表明一个可能的成本方面的策略性能。然而，在健康对照还示出的比较，反应时间的两侧，这也可能是由于执行健康对照的事实，即一个驱动器小于患者的略有不同。为了测试是否这代表稳定的效果，进行更多的临床试验是需要的。 在病人的A，乙病人介绍病人缺乏补偿的有代表性的结果atory行为和驾驶性能的影响:缺乏代偿扫视运动的盲区导致碰撞的对象中出现的盲区和延长反应时间。然而，在整个驱动器，病人自发开始执行更扫视，右视野，以更大的幅度，从而在更小的碰撞的发生率。 <img alt="图1A" src="/files/ftp_upload/4427/4427fig1A.jpg" /> 图1。患者A 30°，自动阈值视野。 <img alt="图1B" src="/files/ftp_upload/4427/4427fig1B.jpg" /> 图1B。病人B，自动化30℃阈值视野。 <img alt="图2" src="/files/ftp_upload/4427/4427fig2.jpg" /> 图2。 </STRONG&gt;分布的注视屏幕上的病人A，乙病人和健康对照。 <img alt="图3" src="/files/ftp_upload/4427/4427fig3.jpg" /> 图3。反应时间出现在各种怪癖的视野，患者A，乙病人和健康对照的对象。 1 tempomat实施，各年龄组之间的反应时间，以保证可比性，因为它是已知的较旧的驱动器降低速度，一个可能的补偿机制7。 2模拟疾病被描述为恶心，出汗，头晕，坚持在驾驶的。有不同的数据出现的频率范围从9％提高到37％，因年龄而异，因为它发生更容易在老人8，9，10。充分的准备和实践驱动器足够长的时间对每一个人进行适当的adjus黄贤金，，减少模拟疾病的机会。 3每页驱动器有4个野猪和4球编程为接近从每一侧的道路，在两个不同的偏心，在直链部分的当然，在不同的时间间隔的过程，以防止测试习惯。触发对象的外观由当事人通过流量点的道路上。

<ol>
	<li>Bouwmeester, L., Heutink, J., Lucas, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+visual+training+for+patients+with+visual+field+defects+due+to+brain+damage:+a+systematic+review.">The effect of visual training for patients with visual field defects due to brain damage: a systematic review.</a> Journal of Neurology, Neurosurgery, and Psychiatry. 78, 555-564 (2007).</li><li>Roth, T., Sokolov, A. N., Messias, A., Roth, P., Weller, M., Trauzettel-Klosinski, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparing+explorative+saccade+and+flicker+training+in+hemianopia.+A+randomized+controlled+study.">Comparing explorative saccade and flicker training in hemianopia. A randomized controlled study.</a> Neurology. 72, 324-331 (2009).</li><li>Freedman, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordination+of+the+eyes+and+head+during+visual+orienting.">Coordination of the eyes and head during visual orienting.</a> Experimental brain research. 190, 369-387 (2008).</li><li>Martin, T., Riley, M. E., Kelly, K. N., Hayhoe, M., Huxlin, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visually-guided+behavior+of+homonymous+hemianopes+in+a+naturalistic+task.">Visually-guided behavior of homonymous hemianopes in a naturalistic task.</a> Vision Research. 47, 3434-3446 (2007).</li><li>Hayhoe, M. M., Ballard, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye+movements+in+natural+behavior.">Eye movements in natural behavior.</a> Trends in Cognitive Sciences. 9, 188-194 (2005).</li><li>Wood, J. M., McGwin, G., Elgin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemianopic+and+quadrantanopic+field+loss,+eye+and+head+movements,+and+driving.">Hemianopic and quadrantanopic field loss, eye and head movements, and driving.</a> Investigative ophthalmology &#38; visual science. 52, 1220-1225 (2011).</li><li>Cantin, V., Lavalli re, M., Simoneau, M., Teasdale, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mental+workload+when+driving+in+a+simulator:+effects+of+age+and+driving+complexity.">Mental workload when driving in a simulator: effects of age and driving complexity.</a> Accident; analysis and prevention. 41, 763-771 (2009).</li><li>Brooks, J. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simulator+sickness+during+driving+simulation+studies.">Simulator sickness during driving simulation studies.</a> Accident; analysis and prevention. 42, 788-796 (2010).</li><li>Allen, R. W., Park, G. D., Fiorentino, D., Rosenthal, T. J., Cook, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+simulator+sickness+as+a+function+of+age+and+gender.">Analysis of simulator sickness as a function of age and gender.</a> , (2006).</li><li>Liu, L., Watson, B., Miyazaki, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VR+for+the+Elderly:+Quantitative+and+Qualitative+Differences+in+Performance+with+a+Driving+Simulator.">VR for the Elderly: Quantitative and Qualitative Differences in Performance with a Driving Simulator.</a> Cyberpsychol. Behav. 2, 567-577 (1999).</li><li>Einh&#228;user, W., Moeller, G. U., Schumann, F., Conradt, J., Vockeroth, J., Bartl, K., Schneider, E., K&#246;nig, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye-Head+Coordination+during+Free+Exploration+in+Human+and+Cat.+Basic+and+Clinical+Aspects+of+Vertigo+and+Dizziness.">Eye-Head Coordination during Free Exploration in Human and Cat. Basic and Clinical Aspects of Vertigo and Dizziness.</a> Ann. N.Y. Acad. Sci. 1164, 353-366 (2009).</li><li>Bowers, A. R., Mandel, A. J., Goldstein, R. B., Peli, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Driving+with+Hemianopia,+I:+Detection+Performance+in+a+Driving+Simulator.">Driving with Hemianopia, I: Detection Performance in a Driving Simulator.</a> Investigative Ophthalmology and Visual Science. 50, 5137-5147 (2009).</li><li>Cockelbergh, T. R. M., Brouwer, W. H., Cornelissen, F. W., van Wolffelaar, P., Kooijman, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effect+of+Visual+Field+Defects+on+Driving+Performance.">The Effect of Visual Field Defects on Driving Performance.</a> Archives of Ophthalmology. 120, 1509-1516 (2002).</li></ol>

没有利益冲突的声明。

新成立的方法，使考试的的视觉探索行为的中风引起的视野缺损的患者。测试设计还提供了一个直接的方法来评估补偿的注视行为:通过将左眼图像上叠加考官可​​以想像的注视行为的病人在测试过程中会话。因此，它可以让一个非常快速和即时评估患者是否采取了补偿的注视行为。它也可以让患者知道它的覆盖眼图像在屏幕上移动的注视反馈工具，可视化的凝视运动。头运动补偿注视行为的作用目前还不清楚。在我们的对照组头运动在老年人中更为常见。比例健康对照组进行头部运动。当测试来看是更广泛的领域比我们的设置时，头部运动可以发挥更大的作用。因此我们不能identif的Ÿ头部运动的代偿的注视行为在我们的病人中的一部分。然而，越来越多的患者需要进行检查，以澄清头运动补偿行为的作用。 这项研究的限制有以下几种:复检成为必要在某些个体，由于在整个驱动器的眼跟踪器的垂直偏移。对象自然出现沿道路，而不是在一个固定偏心所引发的视线位置。然而目前的视线位置关系的对象时，被认为是解释的反应时间。 视野缺损的患者进行了测试前在模拟和真实的驾驶设置:Bowers 等12和13 Cockelbergh 等。在驾驶模拟器进行了研究和证明患者与健康对照组相比，驾驶性能较差。不过，他们并没有录得眼睛和头部运动和个体差异，可能会ŇOT相关的视觉探索行为。 Wood 等人。在现实生活中的情况下进行测试，并建立了一个评估的驾驶性能与视野缺损的患者。头部和眼睛运动进行了分析和测试后得分由两个独立的研究人员通过视频，从而与可靠性评估者间交易。然而，他们没有固定的持续时间，扫视，和头部动作和评估依赖于认证的驾驶康复专家提供了定量分析。我们与模拟驾驶的优势是方便，快捷的评估在临床上，记录良好定义的参数的眼睛和头部动作以及反应时间。这是可以控制的水平会分散人的注意力，使每个驱动器类似的驱动的情况下，标准化的路线和条件允许的可比性。罗斯表明，SCT提高了搜索的行为，就盲目的景象自然的搜寻Ĥ任务。通过调整在驾驶课程分心的水平，这将有可能证明，如果在哪一级，代偿行为与工作量增加失败。比较模拟真实的驾驶研究，教它似乎是适当的补偿行为在模拟环境中，暴露病人到真正的驾驶情况下，作为第二个步骤。特别是因为后者使驾驶安全性评价。 在未来，我们希望包括不同程度的代偿行为分析扫视，振幅和分布的表征。这可以帮助提供更多的个人康复计划调整，以病人目前的补偿行为。其次，乙病人发现的补偿策略的自发采用的，我们作为一个可能的工具来测试的设计，康复的目的不只是作为一个诊断测试的设计，但也提供特殊培训的模拟驾驶，指示日Ë病人进行代偿扫视行为。结合即时的注视行为的可视化表示眼睛覆盖图片的目光，这可能会提供一种反馈机制，出现了注意力进行补偿的策略。

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 产品编号 </td><td> 评论 </td></tr><tr><td>驾驶模拟器软件SILAB的</td><td>维尔茨堡交通科学研究所有限公司（WIVW） </td><td></td><td> <a href="http://www.wivw.de/index.php.en" target="_blank">http://www.wivw.de/index.php.en</a> </td></tr><tr><td> EyeSeeCam </td><td>慕尼黑的医院 临床神经科学</td><td></td><td> <a href="http://eyeseecam.com" target="_blank">http://eyeseecam.com</a> </td></tr><tr><td></td><td></td><td></td><td>估计成本及建立20000欧元，3个月的时间。 </td></tr></tbody></table>

driving simulation in the clinic: testing visual exploratory behavior in daily life activities in patients with visual field defects

患同向偏盲梗死后大脑后动脉（PCA）的患者在日常生活中报告了不同程度的约束，尽管类似的视觉障碍。我们假设，这可能是由于补偿策略，如改变视觉扫描行为变量的发展。扫描代偿治疗（SCT）研究旁边的视力恢复治疗梗死后的视觉训练的一部分。 SCT的学习，使较大的眼球运动的盲区，扩大了可视化搜索领域，这已被证明是最有用的策略，不仅在自然搜索的任务，但也掌握日常生活活动2。然而，在日常临床工作中，它是很难确定的补偿行为的个人水平和培训效果，因为它需要在头奔放的条件下测量眼球运动。研究表明，奔放的头部运动ALTER的视觉探索行为相比，头克制的实验室条件下，3。 Martin等。4，霍等[5]表明，在实验室环境中表现出的行为不能很容易被分配到的自然条件。因此，我们的目标是建立一个研究组，在实际测试的情况下迅速发现不同的代偿性动眼神经战略:患者在临床环境中驾驶模拟器进行测试。软件SILAB（ 维尔茨堡研究所为交通科学GmbH公司（WIVW）），使用程序复杂程度不同的驾驶情况下，记录驱动器的性能。该软件相结合，与头戴式红外线视频瞳孔跟踪，记录头和眼睛运动（EyeSeeCam德国慕尼黑大学医院，临床神经科学）。 驾驶模拟器和定位在病人的定位，调整和校准的相机是DEM的梅雨期。说明在本试验研究和补偿策略及健康对照病人的典型表现。指示受试者凝视在屏幕上位于由动态覆盖图片，并通过分析该数据驱动器本身的过程中，不同的动眼神经的行为（眼和头部运动的频率和振幅的）进行评估非常迅速。补偿注视行为的患者与健康对照的驾驶性能，而没有补偿行为的病人显着的性能更坏。的眼睛和头部的运动行为，以及驾驶性能进行了讨论对于不同的动眼神经战略和在更广泛的范围内，就可能整个测试环节的康复潜力和影响培训效果的数据。

Watch this Scientific Journal Video about Driving Simulation in the Clinic: Testing Visual Exploratory Behavior in Daily Life Activities in Patients with Visual Field Defects at JoVE.com

Driving Simulation in the Clinic: Testing Visual Exploratory Behavior in Daily Life Activities in Patients with Visual Field Defects

患者中风后的报告在日常生活中不同的约束最有可能是由于变量的补偿策略，这是很难区分在临床常规视觉赤字。我们提出了一个临床的设置可以测量不同的代偿头位和眼动战略，并评估其对驾驶性能的影响。

驾驶模拟诊所:测试视觉与视野缺损的患者在日常活动中的探索行为

Patients suffering from homonymous hemianopia after infarction of the posterior cerebral artery (PCA) report different degrees of constraint in daily ...

driving-simulation-clinic-testing-visual-exploratory-behavior-daily

Research

JoVE Journal

Medicine

17.3K 次观看.  Universitätsmedizin Charité. 患者中风后的报告在日常生活中不同的约束最有可能是由于变量的补偿策略，这是很难区分在临床常规视觉赤字。我们提出了一个临床的设置可以测量不同的代偿头位和眼动战略，并评估其对驾驶性能的影响。

视频: Driving Simulation in the Clinic: Testing Visual Exploratory Behavior in Daily Life Activities in Patients with Visual Field Defects

Sequencing of Bacterial Microflora in Peripheral Blood: our Experience with HIV-infected Patients

The healthy gastrointestinal tract is physiologically colonized by a large variety of commensal microbes that influence the development 
	of the humoral and cellular mucosal immune system1,2. 
 Microbiota is shielded from the immune system via a strong mucosal barrier. Infections and antibiotics are known to alter both the normal 
	gastrointestinal tract barrier and the composition of resident bacteria, which may result in possible immune abnormalities3.
HIV causes a breach in the gastrointestinal barrier with progressive failure of mucosal immunity and leakage into the systemic circulation of bacterial bioproducts, such as lipopolysaccharide and 
	bacterial DNA fragments, which contribute to systemic immune activation4-7. Microbial translocation is implicated in HIV/AIDS immunopathogenesis and response to therapy 4,8.
We aimed to characterise the composition of bacteria translocating in peripheral blood of HIV-infected patients. To pursue our aim we set up a PCR reaction for the panbacteric 16S ribosomial gene 
	followed by a sequencing analysis.
Briefly, whole blood from both HIV-infected and healthy subjects is used. Given that healthy individuals present normal intestinal homeostasis no translocation of microflora is expected in 
	these patients. Following whole blood collection by venipuncture and plasma separation, DNA is extracted from plasma and used to perform a broad range PCR reaction for the panbacteric 
	16S ribosomial gene9. Following PCR product purification, cloning and sequencing analyses are performed.

Our experiment will show how to perform a sequencing analysis of bacterial species translocating in peripheral blood of HIV positive patients.

测序外周血中细菌菌群:我们的经验与艾滋病毒感染者

The healthy gastrointestinal tract is physiologically colonized by a large variety of commensal microbes that influence the development 
	of the humoral ...

科研

医学

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio based cues and set within the context of a video game metaphor, users gather relevant spatial information regarding a building's layout. This allows the user to develop an accurate spatial cognitive map of a large-scale three-dimensional space that can be manipulated for the purposes of a real indoor navigation task. After game play, participants are then assessed on their ability to navigate within the target physical building represented in the game. Preliminary results suggest that early blind users were able to acquire relevant information regarding the spatial layout of a previously unfamiliar building as indexed by their performance on a series of navigation tasks. These tasks included path finding through the virtual and physical building, as well as a series of drop off tasks. We find that the immersive and highly interactive nature of the AbES software appears to greatly engage the blind user to actively explore the virtual environment. Applications of this approach may extend to larger populations of visually impaired individuals.

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind.

基于音频的虚拟游戏环境的发展，以帮助盲人的导航技巧

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

A Laser-induced Mouse Model of Chronic Ocular Hypertension to Characterize Visual Defects

Glaucoma, frequently associated with elevated intraocular pressure (IOP), is one of the leading causes of blindness. We sought to establish a mouse model of ocular hypertension to mimic human high-tension glaucoma. Here laser illumination is applied to the corneal limbus to photocoagulate the aqueous outflow, inducing angle closure. The changes of IOP are monitored using a rebound tonometer before and after the laser treatment. An optomotor behavioral test is used to measure corresponding changes in visual capacity. The representative result from one mouse which developed sustained IOP elevation after laser illumination is shown. A decreased visual acuity and contrast sensitivity is observed in this ocular hypertensive mouse. Together, our study introduces a valuable model system to investigate neuronal degeneration and the underlying molecular mechanisms in glaucomatous mice.

Chronic ocular hypertension is induced using laser photocoagulation of the trabecular meshwork in mouse eyes. The intraocular pressure (IOP) is elevated for several months after laser treatment. The decrease of visual acuity and contrast sensitivity of experimental animals are monitored using the optomotor test.

激光诱导的慢性高眼压表征视觉缺陷的小鼠模型

Glaucoma, frequently associated with elevated intraocular pressure (IOP), is one of the leading causes of blindness. We sought to establish a mouse model ...

Detecting Abnormalities in Choroidal Vasculature in a Mouse Model of Age-related Macular Degeneration by Time-course Indocyanine Green Angiography

Indocyanine Green&#160;Angiography (or ICGA) is a technique performed by ophthalmologists to diagnose abnormalities of the choroidal and retinal vasculature of various eye diseases such as age-related macular degeneration (AMD). ICGA is especially useful to image the posterior choroidal vasculature of the eye due to its capability of penetrating through the pigmented layer with its infrared spectrum. ICGA time course can be divided into early, middle, and late phases. The three phases provide valuable information on the pathology of eye problems. Although time-course ICGA by intravenous (IV) injection is widely used in the clinic for the diagnosis and management of choroid problems, ICGA by intraperitoneal injection (IP) is commonly used in animal research. Here we demonstrated the technique to obtain high-resolution ICGA time-course images in mice by tail-vein injection and confocal scanning laser ophthalmoscopy. We used this technique to image the choroidal lesions in a mouse model of age-related macular degeneration. Although it is much easier to introduce ICG to the mouse vasculature by IP, our data indicate that it is difficult to obtain reproducible ICGA time course images by IP-ICGA. In contrast, ICGA via tail vein injection provides high quality ICGA time-course images comparable to human studies. In addition, we showed that ICGA performed on albino mice gives clearer pictures of choroidal vessels than that performed on pigmented mice. We suggest that time-course IV-ICGA should become a standard practice in AMD research based on animal models.

Indocyanine Green Angiography (or ICGA) performed by tail vein injection provides high quality ICGA time course images to characterize abnormalities in mouse choroid.

按时间当然吲哚菁绿血管造影检测脉络膜血管系统异常在年龄相关性黄斑变性的小鼠模型

Indocyanine Green&#160;Angiography (or ICGA) is a technique performed by ophthalmologists to diagnose abnormalities of the choroidal and retinal ...

Dynamic Visual Tests to Identify and Quantify Visual Damage and Repair Following Demyelination in Optic Neuritis Patients

In order to follow optic neuritis patients and evaluate the effectiveness of their treatment, a handy, accurate and quantifiable tool is required to assess changes in myelination at the central nervous system (CNS). However, standard measurements, including routine visual tests and MRI scans, are not sensitive enough for this purpose. We present two visual tests addressing dynamic monocular and binocular functions which may closely associate with the extent of myelination along visual pathways. These include Object From Motion (OFM) extraction and Time-constrained stereo protocols. In the OFM test, an array of dots compose an object, by moving the dots within the image rightward while moving the dots outside the image leftward or vice versa. The dot pattern generates a camouflaged object that cannot be detected when the dots are stationary or moving as a whole. Importantly, object recognition is critically dependent on motion perception. In the Time-constrained Stereo protocol, spatially disparate images are presented for a limited length of time, challenging binocular 3-dimensional integration in time. Both tests are appropriate for clinical usage and provide a simple, yet powerful, way to identify and quantify processes of demyelination and remyelination along visual pathways. These protocols may be efficient to diagnose and follow optic neuritis and multiple sclerosis patients.
In the diagnostic process, these protocols may reveal visual deficits that cannot be identified via current standard visual measurements. Moreover, these protocols sensitively identify the basis of the currently unexplained continued visual complaints of patients following recovery of visual acuity. In the longitudinal follow up course, the protocols can be used as a sensitive marker of demyelinating and remyelinating processes along time. These protocols may therefore be used to evaluate the efficacy of current and evolving therapeutic strategies, targeting myelination of the CNS.

Object From Motion (OFM) and Time-constrained stereo protocols are sensitive tools to identify monocular and binocular dynamic visual function deficits, which are uniquely affected in optic neuritis patients. Furthermore, these tests may be used as quantitative noninvasive tools to assess the extent of myelination along visual pathways.

动态视觉测试，以确定和量化视觉损伤与修复继脱髓鞘视神经炎患者

In order to follow optic neuritis patients and evaluate the effectiveness of their treatment, a handy, accurate and quantifiable tool is required to ...

In Vivo Dynamics of Retinal Microglial Activation During Neurodegeneration: Confocal Ophthalmoscopic Imaging and Cell Morphometry in Mouse Glaucoma

Microglia, which are CNS-resident neuroimmune cells, transform their morphology and size in response to CNS damage, switching to an activated state with distinct functions and gene expression profiles. The roles of microglial activation in health, injury and disease remain incompletely understood due to their dynamic and complex regulation in response to changes in their microenvironment. Thus, it is critical to non-invasively monitor and analyze changes in microglial activation over time in the intact organism. In vivo studies of microglial activation have been delayed by technical limitations to tracking microglial behavior without altering the CNS environment. This has been particularly challenging during chronic neurodegeneration, where long-term changes must be tracked. The retina, a CNS organ amenable to non-invasive live imaging, offers a powerful system to visualize and characterize the dynamics of microglia activation during chronic disorders.
This protocol outlines methods for long-term, in vivo imaging of retinal microglia, using confocal ophthalmoscopy (cSLO) and CX3CR1GFP/+ reporter mice, to visualize microglia with cellular resolution. Also, we describe methods to quantify monthly changes in cell activation and density in large cell subsets (200-300 cells per retina). We confirm the use of somal area as a useful metric for live tracking of microglial activation in the retina by applying automated threshold-based morphometric analysis of in vivo images. We use these live image acquisition and analyses strategies to monitor the dynamic changes in microglial activation and microgliosis during early stages of retinal neurodegeneration in a mouse model of chronic glaucoma. This approach should be useful to investigate the contributions of microglia to neuronal and axonal decline in chronic CNS disorders that affect the retina and optic nerve.

In Vivo Dynamics of Retinal Microglial Activation During Neurodegeneration: Confocal Ophthalmoscopic Imaging and Cell Morphometry in Mouse Glaucoma

Microglia activation and microgliosis are key responses to chronic neurodegeneration. Here, we present methods for in vivo, long-term visualization of retinal CX3CR1-GFP+ microglial cells by confocal ophthalmoscopy, and for threshold and morphometric analyses to identify and quantify their activation. We monitor microglial changes during early stages of age-related glaucoma.

视网膜胶质细胞活化的体内动力学在神经退行性疾病:共聚焦成像眼底及细胞形态计量小鼠青光眼

Microglia, which are CNS-resident neuroimmune cells, transform their morphology and size in response to CNS damage, switching to an activated state with ...

A Simple Mechanical Procedure to Create Limbal Stem Cell Deficiency in Mouse

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with conjunctiva. Patients suffer from recurrent corneal defects, pain, inflammation, and loss of vision.
Previously, a murine model of LSCD was described and compared to two other models. The goal was to produce a consistent mouse model of LSCD that both mimics the phenotype in humans and lasts long enough to make it possible to study the disease pathophysiology and to evaluate new treatments. Here, the technique is described in more detail.
A motorized tool with a rotating burr has been designed to remove the rust rings from the corneal surface or to smooth the pterygium bed in patients. It is a suitable device to create the desired LSCD model. It is a readily available, easy-to-use tool with a fine tip that makes it appropriate for working on small eyes, as in mice. Its application prevents unnecessary trauma to the eye and it does not result in unwanted injuries, as often is the case with chemical injury models. As opposed to a blunt scraper, it removes the epithelium with the basement membrane. In this protocol, the limbal area was abraded two times, and then the whole corneal epithelium was shaved from limbus to limbus. To avoid stroma injury, care was taken not to brush the corneal surface once the epithelium was already removed.

The following article provides an easy, reproducible technique to effectively create a sustainable mouse model of limbal stem cell deficiency (LSCD). This animal model is useful in testing and comparing the efficacy of treatments for limbal stem cell diseases.

一个简单的机械过程来创建在小鼠角膜缘干细胞缺乏症

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with ...

A Method for Quantifying Upper Limb Performance in Daily Life Using Accelerometers

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. It has become important to measure a person's activities in daily life, and not just measure their capacity for activity in the structured environment of a clinic or laboratory. A wearable sensor that is now enabling measurement of daily movement is the accelerometer. Accelerometers are commercially-available devices resembling large wrist watches that can be worn throughout the day. Data from accelerometers can quantify how the limbs are engaged to perform activities in peoples' homes and communities. This report describes a methodology to collect accelerometry data and turn it into clinically-relevant information. First, data are collected by having the participant wear two accelerometers (one on each wrist) for 24 h or longer. The accelerometry data are then downloaded and processed to produce four different variables that describe key aspects of upper limb activity in daily life: hours of use, use ratio, magnitude ratio, and the bilateral magnitude. Density plots can be constructed that visually represent the data from the 24 h wearing period. The variables and their resultant density plots are highly consistent in neurologically-intact, community-dwelling adults. This striking consistency makes them a useful tool for determining if upper limb daily performance is different from normal. This methodology is appropriate for research studies investigating upper limb dysfunction and interventions designed to improve upper limb performance in daily life in people with stroke and other patient populations. Because of its relative simplicity, it may not be long before it is also incorporated in clinical neurorehabilitation practice.

This protocol describes a method to quantify upper limb performance in daily life using wrist-worn accelerometers.

一种使用加速度计量化上肢表现在日常生活的方法

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

眼毛刺角膜上皮磨损与角膜创面愈合的模型研究

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

Integration of Brain Tissue Saturation Monitoring in Cardiopulmonary Exercise Testing in Patients with Heart Failure

Cerebral hypo-oxygenation during rest or exercise negatively impacts the exercise capacity of patients with heart failure with reduced ejection fraction (HF). However, in clinical cardiopulmonary exercise testing (CPET), cerebral hemodynamics is not assessed. NIRS is used to measure cerebral tissue oxygen saturation (SctO2) in the frontal lobe. This method is reliable and valid and has been utilized in several studies. SctO2 is lower during both rest and peak exercise in patients with HF than in healthy controls (66.3 &#177; 13.3% and 63.4 &#177; 13.8% vs. 73.1 &#177; 2.8% and 72 &#177; 3.2%). SctO2 at rest is significantly linearly correlated with peak VO2 (r = 0.602), oxygen uptake efficiency slope (r = 0.501), and brain natriuretic peptide (r = -0.492), all of which are recognized prognostic and disease severity markers, indicating its potential prognostic value. SctO2 is determined mainly by end-tidal CO2 pressure, mean arterial pressure, and hemoglobin in the HF population. This article demonstrates a protocol that integrates SctO2 using NIRS into incremental CPET on a calibrated bicycle ergometer.

This protocol integrated near-infrared spectroscopy into conventional cardiopulmonary exercise testing to identify the involvement of the cerebral hemodynamic response in exercise intolerance in patients with heart failure.

心力衰竭患者心肺运动测试中脑组织饱和监测的集成

Cerebral hypo-oxygenation during rest or exercise negatively impacts the exercise capacity of patients with heart failure with reduced ejection fraction ...