本研究由国家自然科学基金（批准号：82174521），湖南中医药大学研究生创新项目（2022CX109）资助

涉及动物的程序已获得湖南中医药大学机构护理和使用委员会的批准。
1.滚动按摩器的组装
<ol><li>选择由橡胶滚轮、叉架、弹簧、限位挡板、调节夹板、螺钉和亚克力手柄组成的按摩器（图 1）。确保胶辊长 3 厘米，直径 1.6 厘米，弹簧长 3 厘米，直径 0.9 厘米，限位挡板长 3 厘米，宽 2 厘米，手柄长 12 厘米，直径 0.9 厘米。</li>
	<li>力控制：根据文献结果23，发现丁氏滚动法向下的压力约为体重的10%，因此在设计前滚时施加的压力约为大鼠体重的10%（0.2-0.3 N）。通过调整限位挡板的角度，测试称重控制器上按摩器的最大压力约为 0.3 N。这种压力要求满足了大鼠的需要。</li>
	<li>回滚时确保最小压力约为 0.08 N（图 2）。确保压力精确符合丁氏横滚法要求，即正反向力比为3：1。</li>
	<li>在处理之前，要求操作人员使用节拍器软件将滚动频率控制在 140 卷/分钟，并在预实验中练习 3 次以上，以确保操作标准化。</li>
</ol>2. 骨骼肌损伤大鼠模型的建立
<ol><li>将 24 只雄性 Sprague-Dawley 大鼠（体重 200-250 g）随机分为三组，每组 8 只大鼠，包括对照组 （C）、notexin （NTX） 和 notexin 与推拿 （NTX + 推拿），并以标准饮食为食。保持在12小时光照/12小时黑暗循环中，在20-25°C和50%-70%湿度下。</li>
	<li>腹膜内注射用1%戊巴比妥钠（40mg / kg）麻醉，然后用脱毛膏去除右下肢的毛发。去除毛皮后，用生理盐水擦拭残留的乳霜。通过脚趾捏合反应确认充分麻醉。在动物麻醉时涂抹眼药膏滋润眼睛。在整个过程中提供热支持。</li>
	<li>注射前用碘伏消毒液和 75% 酒精交替对右下肢进行皮肤消毒。将浸泡在碘伏消毒液中的棉签接触下肢皮肤中心，然后以打圈的方式向外涂抹。用浸有乙醇的棉签重复上述步骤。</li>
	<li>根据参考方法24建立骨骼肌损伤模型。仅在一条腿上注射 notexin（以防止双重 notexin 注射）。用 30G 针头将 200 μL notexin 溶液（通过在 15 mL 离心管中加入 10 μg notexin 加入 10 mL 生理盐水制备的 10 μg/mL notexin 溶液制备）放入 30G 针头的 1 mL 注射器中，并将 notexin 溶液肌肉注射到腓肠肌中以产生肌肉损伤。</li>
	<li>缓慢注射notexin并等待3秒后再拔出针头（以完全注射）。 注意：Notexin 是一种有毒化学物质，接触开放性伤口后需要立即用大量水冲洗，并在必要时及时就医。</li>
	<li>向对照组的大鼠注射200μL生理盐水。将麻醉的大鼠移至空笼子中，并铺有干净的垫料。注意去除老鼠鼻子和嘴巴周围的衬垫，以保持它们的呼吸畅通。目视观察注射结束时的组织颜色和呼吸频率，直到大鼠恢复足够的意识。</li>
	<li>将大鼠放回笼子，通常饲养24小时。</li>
</ol>3. 推拿疗法
<ol><li>将SD大鼠置于俯卧位，头部用黑布覆盖在用75%酒精消毒的实验平台上，以暴露腓肠肌。不要盖得太紧。</li>
	<li>NTX+推拿组使用推拿按摩器：握住按摩器，将滚轮放在大鼠的腓肠肌上，向前滚动，直到弹簧接触限位挡板。然后缩回力并返回其原始位置，从而进行往复运动（图 3）。</li>
	<li>以每分钟 140 卷的速度滚动按摩器，每次操作 3 分钟。连续3天上午和下午各进行一次按摩。</li>
	<li>每次处理后将大鼠放回笼中，并在最后一次处理后禁食8小时。</li>
</ol>4.实验后从大鼠身上采集血液和组织
<ol><li>根据相关动物实验伦理委员会的要求，通过腹腔注射1%戊巴比妥钠（40mg/kg，腹腔注射）麻醉大鼠。通过脚趾捏合反应确认充分麻醉。采血后通过腹主动脉放血对大鼠实施安乐死。</li>
	<li>注射前用碘伏消毒液和75%酒精交替进行皮肤消毒。将浸泡在聚维酮碘中的棉签触摸腹部皮肤的中心，然后以圆周运动向外涂抹。用浸有乙醇的棉签重复上述步骤。重复消毒 3 次。</li>
	<li>让助手使用两个止血钳来抬起腹部中间的皮肤。作为操作者，使用手术刀切开腹部皮肤和肌肉，从臀部到耻骨联合。</li>
	<li>打开腹腔后，用无菌棉球将肠分开，露出腹后壁的腹主动脉。</li>
	<li>找到腹主动脉，将5mL大鼠血液取入采血管中，并在静置血液1小时后以3000× g 离心10分钟，将血浆放入1.5个微管中。将血浆储存在-80°C。</li>
	<li>用手术剪刀沿着下腹部开口向右下肢的外侧切开皮肤，露出下肢肌肉，用镊子小心地分离筋膜后，切开手术刀以去除完整的腓肠肌。</li>
	<li>用无菌盐水清洗腓肠肌，以去除粘附的毛发和血液。</li>
	<li>将去除的腓肠肌放入含有 4% 多聚甲醛的 15 mL 离心管中。</li>
</ol>5. ELISA检测血浆CK和FABP 3水平
<ol><li>计算并确定一次实验所需的预包装板数量。取出所需的板，将它们放入96孔框架中，将剩余的微孔板放回铝箔袋中密封，并将它们储存在4°C。</li>
	<li>将试剂盒和样品在室温（25-28°C）下平衡120分钟，完全平衡至室温。 注意：试剂盒和样品的平衡至关重要，必须平衡到足够的时间。</li>
	<li>设置标准孔、样品孔和空白孔。向标准孔中加入不同浓度（100、50、25、12.5、6.25、0 ng/mL）的 50 μL CK 或 FABP3 标准品。每个标准重复一次，总共占据12个孔。</li>
	<li>用 40 μL 样品稀释剂（0.8 g NaCl、0.02 g KH 2 PO 4、0.29 gNa 2 HPO 4 12H2O、0.02 g KCl、0.01 g NaN3在 100 mL 双蒸水，pH7.4 中）填充样品孔，然后加入 10 μL 待测样品。重复每个样品一次，总共占用 48 个孔。</li>
	<li>除了位于最后一个样品孔后面两个孔的空白孔外，向每个标准品和样品孔中加入 100 μL HRP 标记的抗人 CK 或 FABP3 抗体（酶标记抗体）。</li>
	<li>用密封膜密封孔，并在37°C水浴或恒温器中孵育60分钟。</li>
	<li>丢弃液体，在吸水纸上拍干，用洗涤液填充每个孔，静置 20 秒，甩掉洗涤液，在吸墨纸上拍干，然后重复洗涤板 5 次（或使用洗板机）。</li>
	<li>向每个孔中加入 50 μL 显色剂溶液 A（20 mg 四甲基联苯胺、10 mL 乙醇溶于 100 mL 双蒸水中）和 50 μL 显色剂溶液 B（0.1 M/L 柠檬酸、0.2 M/L 磷酸二氢钠缓冲液，pH 5.0-5.4）。在37°C下避光15分钟。</li>
	<li>对于标准孔、样品孔和空白孔，向每个孔中加入 50 μL 终止溶液，并在 15 分钟内测量每个孔在 450 nm 处的光密度值。</li>
</ol>6. notexin诱导的大鼠腓肠肌损伤的组织学分析
<ol><li>准备用苏木精和伊红染色的 5 μm 厚石蜡切片，用于光学显微镜检查，如25 中所述。</li>
</ol>7. 图像处理和数据分析
<ol><li>使用分析软件读取和分析成像系统捕获的图像。用鼠标将选定的图像视野移动到屏幕中央，单击 40 倍，然后单击“ 拍摄快照”。</li>
	<li>在电子表格中记录ELISA的OD值，并使用标准曲线计算样品中的大鼠CK和FABP3水平。</li>
	<li>使用统计分析软件进行统计分析。将测量值表示为平均值±标准差 （<img alt="Equation 1" src="/files/ftp_upload/65820/65820eq01.jpg" style="margin: auto;">），并通过单因素方差分析分析组间比较，方差均匀时采用 LSD 检验，方差不均匀时采用 Tamhane T2 方法。当 p 值小于 0.05 时，该差异被认为具有统计学意义。</li>
</ol>

基于 Ding's Roll 操作治疗大鼠模型肌肉损伤的按摩器

<ol>
	<li>Lempainen, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+anterior+thigh+injuries+in+soccer+players:+practical+guide.">Management of anterior thigh injuries in soccer players: practical guide.</a> BMC Sports Science, Medicine and Rehabilitation. 14 (1), 41 (2022).</li><li>Bayer, M. L., Mackey, A., Magnusson, S. P., Krogsgaard, M. R., Kj&#230;r, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+acute+muscle+injuries+(in+Danish).">Treatment of acute muscle injuries (in Danish).</a> Ugeskrift for Laeger. 181 (8), V11180753 (2019).</li><li>Serner, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progression+of+Strength,+Flexibility,+and+Palpation+Pain+During+Rehabilitation+of+Athletes+with+Acute+Adductor+Injuries:+A+Prospective+Cohort+Study.">Progression of Strength, Flexibility, and Palpation Pain During Rehabilitation of Athletes with Acute Adductor Injuries: A Prospective Cohort Study.</a> The Journal of Orthopaedic and Sports Physical Therapy. 51 (3), 126-134 (2021).</li><li>Gozubuyuk, O. B., Koksal, C., Tasdemir, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rehabilitation+of+a+patient+with+bilateral+rectus+abdominis+full+thickness+tear+sustained+in+recreational+strength+training:+a+case+report.">Rehabilitation of a patient with bilateral rectus abdominis full thickness tear sustained in recreational strength training: a case report.</a> Physiotherapy Theory and Practice. 38 (13), 3216-3225 (2022).</li><li>Hotfiel, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+Conservative+Treatment+and+Management+Strategies+of+Skeletal+Muscle+Injuries.">Current Conservative Treatment and Management Strategies of Skeletal Muscle Injuries.</a> Zeitschrift f&#252;r Orthop&#228;die und Unfallchirurgie. 154 (3), 245-253 (2016).</li><li>de Sire, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+Treatment+for+Acute+Traumatic+Musculoskeletal+Pain+in+Athletes.">Pharmacological Treatment for Acute Traumatic Musculoskeletal Pain in Athletes.</a> Medicina. 57 (11), 1208 (2021).</li><li>Connor, D. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapeutic+potential+of+exosomes+in+rotator+cuff+tendon+healing.">Therapeutic potential of exosomes in rotator cuff tendon healing.</a> Journal of Bone and Mineral Metabolism. 37 (5), 759-767 (2019).</li><li>Martins, A. L. L., Giorno, L. P., Santos, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+Engineering+Applied+to+Skeletal+Muscle:+Strategies+and+Perspectives.">Tissue Engineering Applied to Skeletal Muscle: Strategies and Perspectives.</a> Bioengineering. 9 (12), 744 (2022).</li><li>Horgan, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clouds+across+the+new+dawn+for+clinical,+diagnostic+and+biological+data:+accelerating+the+development,+delivery+and+uptake+of+personalized+medicine.">Clouds across the new dawn for clinical, diagnostic and biological data: accelerating the development, delivery and uptake of personalized medicine.</a> Diagnosis. , (2023).</li><li>Urits, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Comprehensive+Review+of+Alternative+Therapies+for+the+Management+of+Chronic+Pain+Patients:+Acupuncture,+Tai+Chi,+Osteopathic+Manipulative+Medicine,+and+Chiropractic+Care.">A Comprehensive Review of Alternative Therapies for the Management of Chronic Pain Patients: Acupuncture, Tai Chi, Osteopathic Manipulative Medicine, and Chiropractic Care.</a> Advances in Therapy. 38 (1), 76-89 (2021).</li><li>Lee, N. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chuna+(or+Tuina)+Manual+Therapy+for+Musculoskeletal+Disorders:+A+Systematic+Review+and+Meta-Analysis+of+Randomized+Controlled+Trials.">Chuna (or Tuina) Manual Therapy for Musculoskeletal Disorders: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.</a> Evidence-based Complementary and Alternative Medicine: eCAM. 2017, 8218139 (2017).</li><li>Xie, J., Deng, D. X., Chen, Y., Peng, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress+in+the+intervention+of+massage+techniques+on+skeletal+muscle+injury.">Progress in the intervention of massage techniques on skeletal muscle injury.</a> Hunan Journal of Traditional Chinese Medicine. 34 (04), 199-201 (2018).</li><li>Yuan, Y., Zhang, H., Zhang, G. H., Xue, X. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Research+progress+on+microstructure+changes+and+rehabilitation+treatment+of+exercise-induced+skeletal+muscle+injury.">Research progress on microstructure changes and rehabilitation treatment of exercise-induced skeletal muscle injury.</a> Massage and Rehabilitation Medicine. 14 (6), 29-33 (2023).</li><li>Zhao, 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+Establishment+of+Famous+Tuina+Master+Ding+Jifeng+and+Wei+Fa+-+Commemorating+the+100th+Anniversary+of+Mr.+Ding+Jifeng's+Birthday.">The Establishment of Famous Tuina Master Ding Jifeng and Wei Fa - Commemorating the 100th Anniversary of Mr. Ding Jifeng's Birthday.</a> Traditional Chinese Medicine Culture. 9 (6), 18-21 (2014).</li><li>Hardy, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+Study+of+Injury+Models+for+Studying+Muscle+Regeneration+in+Mice.">Comparative Study of Injury Models for Studying Muscle Regeneration in Mice.</a> PloS one. 11 (1), e0147198 (2016).</li><li>Plant, D. R., Colarossi, F. E., Lynch, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notexin+causes+greater+myotoxic+damage+and+slower+functional+repair+in+mouse+skeletal+muscles+than+bupivacaine.">Notexin causes greater myotoxic damage and slower functional repair in mouse skeletal muscles than bupivacaine.</a> Muscle &amp; Nerve. 34 (5), 577-585 (2006).</li><li>Tierney, M. T., Sacco, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inducing+and+Evaluating+Skeletal+Muscle+Injury+by+Notexin+and+Barium+Chloride.">Inducing and Evaluating Skeletal Muscle Injury by Notexin and Barium Chloride.</a> Methods in Molecular Biology. 1460, 53-60 (2016).</li><li>Torres-Ruiz, J., Alcal&#225;-Carmona, B., Alejandre-Aguilar, R., G&#243;mez-Mart&#237;n, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammatory+myopathies+and+beyond:+The+dual+role+of+neutrophils+in+muscle+damage+and+regeneration.">Inflammatory myopathies and beyond: The dual role of neutrophils in muscle damage and regeneration.</a> Frontiers in Immunology. 14, 1113214 (2023).</li><li>Tu, H., Li, Y. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammation+balance+in+skeletal+muscle+damage+and+repair.">Inflammation balance in skeletal muscle damage and repair.</a> Frontiers in Immunology. 14, 1133355 (2023).</li><li>Castro, C., Gourley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis+and+treatment+of+inflammatory+myopathy:+issues+and+management.">Diagnosis and treatment of inflammatory myopathy: issues and management.</a> Therapeutic Advances in Musculoskeletal Disease. 4 (2), 111-120 (2012).</li><li>Dabby, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymptomatic+or+minimally+symptomatic+hyperCKemia:+histopathologic+correlates.">Asymptomatic or minimally symptomatic hyperCKemia: histopathologic correlates.</a> The Israel Medical Association Journal: IMAJ. 8 (2), 110-113 (2006).</li><li>Khodabukus, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue-Engineered+Human+Myobundle+System+as+a+Platform+for+Evaluation+of+Skeletal+Muscle+Injury+Biomarkers.">Tissue-Engineered Human Myobundle System as a Platform for Evaluation of Skeletal Muscle Injury Biomarkers.</a> Toxicological Sciences. 176 (1), 124-136 (2020).</li><li>Zhou, X. W., Jin, W. D., Zhu, L., Liu, X. H., Zhou, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+observation+on+the+influence+of+different+frequency,+intensity+and+action+time+of+Ding+rolling+manipulation+on+hemodynamics.">Experimental observation on the influence of different frequency, intensity and action time of Ding rolling manipulation on hemodynamics.</a> Shanghai Journal of Traditional Chinese Medicine. (06), 42-44 (1998).</li><li>Pablos, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+Effects+of+Foam+Rolling+against+Inflammation+and+Notexin+Induced+Muscle+Damage+in+Rats.">Protective Effects of Foam Rolling against Inflammation and Notexin Induced Muscle Damage in Rats.</a> International Journal of Medical Sciences. 17 (1), 71-81 (2017).</li><li>Wisner, L., Larsen, B., Maguire, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancing+Tumor+Content+through+Tumor+Macrodissection.">Enhancing Tumor Content through Tumor Macrodissection.</a> Journal of Visualized Experiments: JoVE. (180), e62961 (2022).</li><li>Deng, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contusion+concomitant+with+ischemia+injury+aggravates+skeletal+muscle+necrosis+and+hinders+muscle+functional+recovery.">Contusion concomitant with ischemia injury aggravates skeletal muscle necrosis and hinders muscle functional recovery.</a> Experimental Biology and Medicine. 247 (17), 1577-1590 (2022).</li><li>Dobek, G. L., Fulkerson, N. D., Nicholas, J., Schneider, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+of+muscle+crush+injury+of+the+legs.">Mouse model of muscle crush injury of the legs.</a> Comparative Medicine. 63 (3), 227-232 (2013).</li><li>Armstrong, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sildenafil+citrate+protects+skeletal+muscle+of+ischemia-reperfusion+injury:+immunohistochemical+study+in+rat+model.">Sildenafil citrate protects skeletal muscle of ischemia-reperfusion injury: immunohistochemical study in rat model.</a> Acta Ciru&#769;rgica Brasileira. 28 (4), 282-287 (2013).</li></ol>

作者没有什么可透露的。

在这里，我们描述了推拿治疗大鼠骨骼肌损伤的方案，然后分析了治疗后骨骼肌损伤的程度，以验证该方法的有效性。值得注意的是，大鼠骨骼肌损伤模型，包括但不限于药物诱导（notexin、布比卡因）16、钝挫伤26、挤压 27 和缺血再灌注28，可以用推拿进行干预。通过HE染色观察组织病理学变化和ELISA检测确定骨骼肌损伤标志?...

肌肉损伤是临床和日常生活中常见的创伤性损伤，由外部打击（挫伤）或肌纤维慢性过度拉伤（拉伤）等引起，导致肌肉功能障碍和疼痛，甚至严重影响患者的生活质量1。急性劳损后尽早开始康复是缩短恢复运动时间的关键2 和减轻疼痛 3,4。在现代西医中，肌肉损伤的临床急救遵循休息、冰敷、加压和抬高 （RICE） 原则，以阻止肌肉组织有害出血5 和非甾体抗炎药缓解疼痛6。外泌体7 和组织工程8 等新疗法的发现成为骨骼肌疾病的潜在治疗策略，弥补了先前药物治疗的不足。然而，它也会增加患者的治疗成本，使他们承受巨大的经济压力9.因此，建议使用替代疗法和补充疗法来治疗肌肉骨骼问题10.推拿作为传统医疗方法在中国临床上被广泛使用，因其疗效好、副作用少而受到患者的欢迎。推拿治疗肌肉骨骼疾病可以减轻疼痛并改善功能11,12,13。上海著名推拿法师丁继峰先生创立了丁氏卷法14。它是一种独特的轧碎技术，受力面积大，力均匀柔和，穿透力强。
不同的动物模型基于不同的病因。它们各有利弊，选择正确、合适的动物模型对基础实验具有重要意义，有助于了解骨骼肌损伤后再生和修复的细胞和分子信号通路，为开发治疗骨骼肌疾病的新疗法奠定基础意义。化学诱导的肌肉损伤模型被广泛使用，骨骼肌注射引起肌纤维坏死并产生可以在 2 周内有效再生的再生区域15。notexin 和布比卡因都会导致肌肉损伤。然而，与布比卡因相比，notexin 对骨骼肌的肌毒性损伤更严重，并且自然功能恢复相对较慢16.药物肌内注射成型不仅时间更短，而且对骨骼肌损伤的效果和程度也可控。这种可量化的控制使成功的成型难度降低15,17。
炎症反应是一种重要的生物学反应，已在肌病的背景下进行了广泛研究18,19。在骨骼肌损伤的早期阶段，肌纤维坏死会破坏局部肌肉稳态，许多炎症细胞浸润损伤部位，分泌许多促炎细胞因子19。肌酸激酶 （CK） 是用于评估骨骼肌损伤的传统血清生物标志物。然而，它缺乏组织特异性20 和敏感性21，这限制了其评估药物诱导的肌肉损伤程度和间接报告损伤后肌肉恢复程度的能力。包括脂肪酸结合蛋白 3 （FABP3） 在内的新型生物标志物最近在骨骼肌损伤的啮齿动物模型中显示出相对较高的组织特异性和敏感性。FABP3 是一个结合蛋白家族，主要在心脏和骨骼肌细胞中表达，与脂肪酸代谢、转运和信号传导有关22。因此，我们选择了两种生物标志物CK和FABP3的组合，以评估notexin诱导的骨骼肌损伤和治疗后的恢复程度。
在啮齿类动物中，肌肉较浅，皮肤面积较小，这也决定了啮齿类动物按摩的各种参数不会与人类相同，例如在动物治疗中，按摩治疗师应使用丁氏滚动法以较小的力气对待它们，并且可能由于受伤部位的尺寸较小，不利于该技术的操作， 这最终会导致按摩效果的降低。因此，本实验利用自制的符合丁氏滚动法特点的滚动按摩器，对notexin诱导的大鼠骨骼肌损伤模型的治疗效果进行干预和评价，有助于在实验动物研究中标准化推拿参数，以深入研究推拿的分子作用机制。 一种中医治疗方法，用于肌肉骨骼疾病。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 mL syringe</td>
			<td>JIANGXI FENGLIN</td>
			<td>20220521</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 microtubes&#160;</td>
			<td>Servicebio</td>
			<td>EP-150X-J</td>
			<td></td>
		</tr>
		<tr>
			<td>15 mL centrifuge tube</td>
			<td>Servicebio</td>
			<td>EP-1501-J</td>
			<td></td>
		</tr>
		<tr>
			<td>30G needle</td>
			<td>CONPUVON</td>
			<td>220318</td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL blood collection tube</td>
			<td>Servicebio</td>
			<td>QX0023</td>
			<td></td>
		</tr>
		<tr>
			<td>Acrylic handle</td>
			<td>Guangdong Guangxingwang Plastic Materials Co., Ltd</td>
			<td>65643645</td>
			<td></td>
		</tr>
		<tr>
			<td>Adjustment splint</td>
			<td>CREROMEM</td>
			<td>20220729</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Swab</td>
			<td>INOHV</td>
			<td>22080215</td>
			<td></td>
		</tr>
		<tr>
			<td>Enzyme-labeled Instrument</td>
			<td>Rayto</td>
			<td>RT-6100&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>INOHV</td>
			<td>211106</td>
			<td></td>
		</tr>
		<tr>
			<td>Fork holder</td>
			<td>Yongkang Kangzhe Health Technology Co., Ltd</td>
			<td>JL001</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair removal cream</td>
			<td>Veet, France</td>
			<td>LOTC190922002</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin dyeing solution set</td>
			<td>Wuhan Google Biotech</td>
			<td>G1005</td>
			<td></td>
		</tr>
		<tr>
			<td>Imaging system&#160;</td>
			<td>Nikon, Japan</td>
			<td>Nikon DS-U3</td>
			<td></td>
		</tr>
		<tr>
			<td>IODOPHOR disfecting solution</td>
			<td>Hale&#38;Hearty</td>
			<td>20221205</td>
			<td></td>
		</tr>
		<tr>
			<td>Light microscope</td>
			<td>Nikon, Japan</td>
			<td>Nikon Eclipse E100</td>
			<td></td>
		</tr>
		<tr>
			<td>Limit baffle</td>
			<td>CREROMEM</td>
			<td>20220724</td>
			<td></td>
		</tr>
		<tr>
			<td>Notexin</td>
			<td>Latoxan S.A.S.</td>
			<td>L8104-100UG</td>
			<td></td>
		</tr>
		<tr>
			<td>Pentobarbital sodium</td>
			<td>Merck KGaA</td>
			<td>P3761</td>
			<td></td>
		</tr>
		<tr>
			<td>Rat creatine kinase (CK) ELISA kit</td>
			<td>LunChangShuoBiotech</td>
			<td>YD-35237</td>
			<td></td>
		</tr>
		<tr>
			<td>Rat fatty acid-binding protein 3 (FABP3) ELISA kit</td>
			<td>LunChangShuoBiotech</td>
			<td>YD-35730</td>
			<td></td>
		</tr>
		<tr>
			<td>Rubber roller</td>
			<td>Hebei Mgkui Chemical Technology Co.,Ltd</td>
			<td>202207</td>
			<td></td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>Weiyan Hardware</td>
			<td>B05Z122</td>
			<td></td>
		</tr>
		<tr>
			<td>Sprague Dawley rats</td>
			<td>Hunan Slake Kingda Laboratory Animal Co.</td>
			<td>SYXK2019-0009</td>
			<td></td>
		</tr>
		<tr>
			<td>Spring</td>
			<td>Bingzhang Hardware</td>
			<td>TH001</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical blade</td>
			<td>Covetrus</td>
			<td>#23</td>
			<td></td>
		</tr>
		<tr>
			<td>Weigh controller</td>
			<td>Iyoys</td>
			<td>HY-XSQ</td>
			<td></td>
		</tr>
	</tbody>
</table>

mimicking ding's roll method on notexin-induced muscle injury in rats

丁氏翻滚法是中国传统按摩诊所中最常用的推拿法之一，也是中国当代推拿法中最有影响力的推拿法之一。它以单指禅宗流派中常用的传统卷法为基础，命名为丁氏卷法。由于其抗炎和促进血液循环的作用，丁氏滚动法对肌病有很好的治疗效果。由于施加在人体皮肤上的力面积很大，丁氏滚动法在皮肤面积小的实验动物（如大鼠和兔子）上具有挑战性。此外，推拿应用于人体的强度与应用于实验动物的强度不同，因此在实验过程中可能会发生强度过高或过低而无法达到推拿的治疗效果。该实验旨在根据 Ding 的滚动操作参数（强度、频率、推拿持续时间）创建一种适合大鼠的简单按摩器。该装置可以标准化动物实验中的操作，并减少由于主观因素而施加到不同动物身上的推拿力的变化。建立大鼠骨骼肌损伤模型，采用血浆损伤标志物肌酸激酶（CK）和脂肪酸结合蛋白3（FABP3）评价推拿对骨骼肌损伤的治疗效果。结果表明，该推拿按摩器可降低CK和FABP3的表达水平，减缓骨骼肌损伤的程度。因此，本文介绍的推拿按摩器，模仿丁氏滚动法，有助于实验研究中推拿操作的标准化，对后续推拿治疗肌病分子机制的研究有很大帮助。

为了观察损伤后大鼠骨骼肌的形态学特性，用苏木精和伊红对腓肠肌进行染色，并用方案中描述的分析软件读取染色图像，每组8只大鼠。在notexin诱导的腓肠肌损伤大鼠（NTX组）中，许多肌肉细胞破裂、萎缩、坏死和排列不规则。受影响区域周围也有中性粒细胞和淋巴细胞的高浸润（图4B）。然而，在用滚动按摩器治疗推拿后，NTX+推拿组肌肉细胞的病理状况有所改善，破裂?...

Watch this Scientific Journal Video about Performing Tuina Massage in Rats Using a Self-Made Massager at JoVE.com

作者聚焦：使用自制按摩器对大鼠进行推拿按摩 

该协议描述了一种模仿Ding滚动方法的简单装置，建立了骨骼肌损伤的大鼠模型，并使用苏木精-伊红染色观察受损组织的病理学和酶联免疫吸附测定法来检测血清损伤标志物的变化。

模仿丁氏滚动法治疗大鼠肌损伤

Ding's roll method is one of the most commonly used manipulations in traditional Chinese massage (Tuina) clinics and one of the most influential ...

我们的研究范围是推拿治疗肌肉骨骼疾病的生物学机制。在这里，我们试图制作自制的推拿装置，以标准化并有效地对实验动物进行推拿治疗。我们正在通过创建一个可以模拟大鼠滚动操作的按摩器来解决无法成对进行这种滚动操作的问题，并验证按摩器对肌肉损伤大鼠的有效性。在未来的标准化研究中，我们将重点关注滚动按摩器对不同参数下大鼠骨骼肌损伤的疗效问题。

mimicking-ding-s-roll-method-on-notexin-induced-muscle-injury-in-rats

Research

JoVE Journal

Medicine

Mimicking Ding's Roll Method on Notexin-Induced Muscle Injury in Rats

945 次观看.  Hunan University of Chinese Medicine. 我们的研究范围是推拿治疗肌肉骨骼疾病的生物学机制。在这里，我们试图制作自制的推拿装置，以标准化并有效地对实验动物进行推拿治疗。我们正在通过创建一个可以模拟大鼠滚动操作的按摩器来解决无法成对进行这种滚动操作的问题，并验证按摩器对肌肉损伤大鼠的有效性。在未来的标准化研究中，我们将重点关注滚动按摩器对不同参数下大鼠骨骼肌损伤的疗效?...

视频: 作者聚焦：使用自制按摩器对大鼠进行推拿按摩 

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

在大鼠脊髓损伤后的急性和慢性触觉测试

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

科研

医学

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

挫伤模型脊髓严重损伤的保护作用

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

评价呼吸肌慢性脊髓损伤个人使用呼吸电机控制评估（RMCA）

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

无线电遥测系统，以监测大鼠脊髓横断和胚胎神经干细胞移植在心血管功能

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Early Detection of Drug-Induced Renal Hemodynamic Dysfunction Using Sonographic Technology in Rats

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is estimated to contribute to 19- 25% of all clinical cases of acute kidney injury (AKI) in critically ill patients. AKI detection has historically relied on metrics such as serum creatinine (sCr) or blood urea nitrogen (BUN) which are demonstrably inadequate in full assessment of nephrotoxicity in the early phase of renal dysfunction. Currently, there is no robust diagnostic method to accurately detect hemodynamic alteration in the early phase of AKI while such alterations might actually precede the rise in serum biomarker levels. Such early detection can help clinicians make an accurate diagnosis and help in in decision making for therapeutic strategy. Rats were treated with Cisplatin to induce AKI. Nephrotoxicity was assessed for six days using high-frequency sonography, sCr measurement and upon histopathology of kidney. Hemodynamic evaluation using 2D and Color-Doppler images were used to serially study nephrotoxicity in rats, using the sonography. Our data showed successful drug-induced kidney injury in adult rats by histological examination. Color-Doppler based sonographic assessment of AKI indicated that resistive-index (RI) and pulsatile-index (PI) were increased in the treatment group; and peak-systolic velocity (mm/s), end-diastolic velocity (mm/s) and velocity-time integral (VTI, mm) were decreased in renal arteries in the same group. Importantly, these hemodynamic changes evaluated by sonography preceded the rise of sCr levels. Sonography-based indices such as RI or PI can thus be useful predictive markers of declining renal function in rodents. From our sonography-based observations in the kidneys of rats that underwent AKI, we showed that these noninvasive hemodynamic measurements may consider as an accurate, sensitive and robust method in detecting early stage kidney dysfunction. This study also underscores the importance of ethical issues associated with animal use in research.

Early stage hemodynamic dysfunction is critical to the development of kidney disease. Yet, detection methodologies are limited. Recent advances in sonography provide a noninvasive, accurate option for early detection of kidney injury. This study outlines a step-by-step, sonographic methodology for detecting kidney dysfunction using a drug-induced nephrotoxicity rat model.

大鼠使用超声技术药物引起的肾血流动力学障碍的早期检测

The kidney normally functions to maintain hemodynamic homeostasis and is a major site of damage caused by drug toxicity. Drug-induced nephrotoxicity is ...

Ultrasonography in Experimental Reproductive Investigations on Rats

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in reproductive medicine. In the past, the study of reproductive system development in rodents has been based on one-time histological examination of excised tissues. Recently, with the development of high-resolution transabdominal ultrasound, high-quality sonography can now be performed to evaluate the reproductive organs of rats, allowing a new method for studying the reproductive system. Images were obtained using a high-resolution ultrasonographic system. Gynecological ultrasonography was performed on 28 eight-week-old non-pregnant rats and 5 pregnant Sprague-Dawley rats. We describe how to recognize organs of the reproductive system and associated structures in typical views during different phases of the estrus cycle. Color flow Doppler was used to measure uterine artery blood flow and evaluate uterine blood flow pattern changes during different stages of pregnancy. We have demonstrated that ultrasound exploration is a useful method for evaluating changes in internal reproductive organs. Its use raises the possibility of conducting additional experiments, including medical or surgical procedures, and provides the ability to monitor sonographic changes to internal organs without sacrificing animals.

This manuscript describes the utility of ultrasound performed on female rats to design experimental models for reproductive and gynecological investigation. A step-by-step explanation of how to perform ultrasonographic evaluation is shown.

大鼠实验性生殖研究中的超声检查

With the development of assisted reproductive technology and the ethical limitations of research on humans, rat animal models have been widely used in ...

Evaluation of Stem Cell Therapies in a Bilateral Patellar Tendon Injury Model in Rats

Regenerative medicine provides novel alternatives to conditions that challenge traditional treatments. The prevalence and morbidity of tendinopathy across species, combined with the limited healing properties of this tissue, have prompted the search for cellular therapies and propelled the development of experimental models to study their efficacy. Umbilical cord matrix-derived mesenchymal stem cells (UCM-MSC) are appealing candidates because they are abundant, easy to collect, circumvent the ethical concerns and risk of teratoma formation, yet resemble primitive embryonic stem cells more closely than adult tissue-derived MSCs. Significant interest has focused on chitosan as a strategy to enhance the properties of MSCs through spheroid formation. This paper details techniques to isolate UCM-MSCs, prepare spheroids on chitosan film, and analyze the effect of spheroid formation on surface marker expression. Consequently, creation of a bilateral patellar tendon injury model in rats is described for in vivo implantation of UCM-MSC spheroids formed on chitosan film. No complication was observed in the study with respect to morbidity, stress rising effects, or tissue infection. The total functional score of the operated rats at 7 days was lower than that of normal rats, but returned to normal within 28 days after surgery. Histological scores of tissue-healing confirmed the presence of a clot in treated defects evaluated at 7 days, absence of foreign body reaction, and progressing healing at 28 days. This bilateral patella tendon defect model controls inter-individual variation via creation of an internal control in each rat, was associated with acceptable morbidity, and allowed detection of differences between untreated tendons and treatments.

This paper describes the preparation and evaluation of umbilical cord matrix-derived mesenchymal stem cells spheroids with a bilateral patellar tendon defect model in a rat. This model was associated with an acceptable morbidity and was found to detect differences between untreated and treated tendons, and between the two treatments tested.

大鼠双侧髌腱损伤模型中干细胞治疗的评价

Regenerative medicine provides novel alternatives to conditions that challenge traditional treatments. The prevalence and morbidity of tendinopathy across ...

Spinal Cord Lateral Hemisection and Asymmetric Behavioral Assessments in Adult Rats

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human Brown-S&#233;quard syndrome&#160;is a common type of incomplete SCI caused by a lesion to one half of the spinal cord which results in paralysis and loss of&#160;proprioception&#160;on the same (or ipsilesional) side as the injury, and loss of pain and temperature sensation on the opposite (or contralesional) side. Adequate methodologies for producing a spinal cord lateral hemisection (HX) and assessing neurological impairments are essential to establish a reliable animal model of Brown-S&#233;quard syndrome. Although lateral hemisection model plays a pivotal role in basic and translational research, standardized protocols for creating such a hemisection and assessing unilateralized function are lacking. The goal of this study is to describe step-by-step procedures to produce a rat spinal lateral HX at the 9th thoracic (T9) vertebral level. We, then, describe a combined behavior scale for HX (CBS-HX) that provides a simple and sensitive assessment of asymmetric neurological performance for unilateral SCI. The CBS-HX, ranging from 0 to 18, is composed of 4 individual assessments which include unilateral hindlimb stepping (UHS), coupling, contact placing, and grid walking. For CBS-HX, the ipsilateral and contralateral hindlimbs are assessed separately. We found that, after a T9 HX, the ipsilateral hindlimb showed impaired behavior function whereas the contralateral hindlimb showed substantial recovery. The CBS-HX effectively discriminated behavioral functions between ipsilateral and contralateral hindlimbs and detected temporal progression of recovery of the ipsilateral hindlimb. The CBS-HX components can be analyzed separately or in combination with other measures when needed. Although we only provided visual descriptions of the surgical procedures and behavioral assessments of a thoracic HX, the principle may be applied to other incomplete SCIs and at other levels of the injury.

Here we describe surgical procedures to produce a reliable spinal cord lateral hemisection (HX) at the 9th thoracic level in adult rats and neurobehavioral assessments designed for detecting asymmetric deficits after such a unilateral injury.

成年大鼠脊髓侧性横断与不对称行为评估

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human ...

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss of relevant amounts of heart muscle mass can lead to the heart failure. Besides heart transplantation, there is no curative treatment option for the end-stage heart failure. In times of organ donor shortage, organ independent treatment modalities are needed. Left-ventricular assist devices are a promising therapy option, however, especially as destination therapy, limited by its side-effects like stroke, infections and bleedings. In recent years, several cardiac repair strategies including stem cell injection, cardiac progenitors or myocardial tissue engineering have been investigated. Recent improvements in cell biology allow for the differentiation of large amounts of cardiomyocytes derived from human induced pluripotent stem cells (iPSC). One of the cardiac repair strategies currently under evaluation is to transplant artificial heart tissue. Engineered heart tissue (EHT) is a three-dimensional in vitro created cardiomyocyte network, with functional properties of native heart tissue. We have created EHT-patches from hiPSC derived cardiomyocytes. Here we present a protocol for the induction of left ventricular myocardial cryoinjury in a guinea pig, followed by implantation of hiPSC derived EHT on the left ventricular wall.

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

几内亚猪模型心肌损伤后 hiPSC-derived 衍生心肌贴片的植入

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

大鼠心肌缺血再灌注后冠状动脉流动储备评价

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...