Name: pseudofracture: an acute peripheral tissue trauma model
Uploaded: 2011-04-18T11:00:00
Description: Watch this Scientific Journal Video about Pseudofracture: An Acute Peripheral Tissue Trauma Model at JoVE.com

资金来源/编号分子生物学失血性休克GM053789

1。仪器及手术现场准备： 所有的实验程序进行使用无菌操作技术。实验区开始之前，必须彻底清洗和消毒。台式，应进行消毒，空气干燥，然后用70％的酒精抹允许。将一个蓝色的手术垫和无菌区，敷在试点工作区。 所有材料和工具，高压灭菌消毒后方可使用。收到无菌注射器和针头。研究者应适当garbed白大褂，口罩，无菌手术手套。 <p class="jove_conte...

<ol>
	<li>DeCamp, M. M., Demling, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Posttraumatic+multisystem+organ+failure.">Posttraumatic multisystem organ failure.</a> JAMA. 260, 530-534 (1988).</li><li>Deitch, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+organ+failure.+Pathophysiology+and+potential+future+therapy.">Multiple organ failure. Pathophysiology and potential future therapy.</a> Ann Surg. 216, 117-134 (1992).</li><li>Carrico, C. J., Meakins, J. L., Marshall, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+organ+failure+syndrome.">Multiple organ failure syndrome.</a> Arch Surg. 121, 196-208 (1986).</li><li>Hauser, C. J., Joshi, P., Jones, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+natural+killer+cell+activity+in+patients+with+fracture/soft+tissue+injury.">Suppression of natural killer cell activity in patients with fracture/soft tissue injury.</a> Arch Surg. 132, 1326-1330 (1997).</li><li>Faist, E., Baue, A. E., Dittmer, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+organ+failure+in+polytrauma+patients.">Multiple organ failure in polytrauma patients.</a> J Trauma. 23, 775-787 (1983).</li><li>Baker, C. C., Oppenheimer, L., Stephens, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+trauma+deaths.">Epidemiology of trauma deaths.</a> Am J Surg. 140, 144-150 (1980).</li><li>Faist, E., Kupper, T. S., Bakeer, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depression+of+cellular+immunity+after+major+injury+its+association+with+posttraumatic+complications+and+its+reversal+with+immunomodulation.">Depression of cellular immunity after major injury its association with posttraumatic complications and its reversal with immunomodulation.</a> Arch Surg. 121, 1000-1005 (1986).</li><li>Lenz, A., Franklin, G. A., Cheadle, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systemic+inflammation+after+trauma.">Systemic inflammation after trauma.</a> Injury. 38, 1336-1345 (2007).</li><li>Flohe, S., Flohe, S. B., Schade, F. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+response+of+severely+injured+patients--influence+of+surgical+intervention+and+therapeutic+impact.">Immune response of severely injured patients--influence of surgical intervention and therapeutic impact.</a> Lang Arch Surg. 392, 639-648 (2007).</li><li>Ayala, A., Wang, P., Ba, Z. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+alterations+in+plasma+IL-6+and+TNF+levels+after+trauma+and+hemorrhage.">Differential alterations in plasma IL-6 and TNF levels after trauma and hemorrhage.</a> Am J Physiol. 260, R167-R171 (1991).</li><li>Kalicke, T., Schlegel, U., Printzen, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+a+standardized+closed+soft+tissue+trauma+on+resistance+to+local+infection.+An+experimental+study+in+rats.">Influence of a standardized closed soft tissue trauma on resistance to local infection. An experimental study in rats.</a> J Ortho Res. 21, 373-378 (2003).</li><li>Kobbe, P., Vodovotz, Y., Kaczorowski, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+fracture+associated+soft+tissue+injury+in+the+induction+of+sytemic+inflammation+and+remote+organ+dysfunction+after+bilateral+femur+fracture.">The role of fracture associated soft tissue injury in the induction of sytemic inflammation and remote organ dysfunction after bilateral femur fracture.</a> J Ortho Trauma. 22, 385-390 (2008).</li><li>Kobbe, P., Vodovotz, Y., Kaczorwoski, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pattern+of+cytokine+release+and+evolution+of+remote+organ+dysfunction+after+bilateral+femur+fracture.">Pattern of cytokine release and evolution of remote organ dysfunction after bilateral femur fracture.</a> Shock. 30, 43-47 (2008).</li><li>Flohe, S. B., Flohe, S., Schade, F. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deterioration+of+the+immune+system+after+trauma:+signals+and+cellular+mechanisms.">Deterioration of the immune system after trauma: signals and cellular mechanisms.</a> Inn. Immun. 14, 333-344 (2008).</li><li>Maier, B., LeFering, R., Lhenert, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+versus+late+onset+of+multiple+organ+failure+is+associated+with+differing+patterns+of+plasma+cytokine+biomarker+expression+and+outcome+after+severe+trauma.">Early versus late onset of multiple organ failure is associated with differing patterns of plasma cytokine biomarker expression and outcome after severe trauma.</a> Shock. 28, 668-674 (2007).</li><li>Mestas, J., Hughes, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Of+mice+not+men:+differences+between+mouse+and+human+immunology.">Of mice not men: differences between mouse and human immunology.</a> J. Immunol. 172, 2731-2738 (2004).</li><li>Hoth, J. J., Wells, J. D., Brownlee, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toll+like+receptor+4-dependent+responses+to+lung+injury+in+a+murine+model+of+pulmonary+contusion.">Toll like receptor 4-dependent responses to lung injury in a murine model of pulmonary contusion.</a> Shock. 31, 376-381 (2009).</li><li>Matsutani, T., Samy, A. n. a. n. t. h. a., Rue, T. S., W, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+prolactin-/-+mice:+effect+of+trauma-hemorrhage+on+splenocyte+functions.">Transgenic prolactin-/- mice: effect of trauma-hemorrhage on splenocyte functions.</a> Am J Physiol Cell Physiol. 288, 1109-1116 (2005).</li><li>Matsutani, T., Samy, A. n. a. n. t. h. a., Kang, T. S., S-C, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+genetic+background+influences+severity+of+immune+responses+following+trauma-hemorrhage.">Mouse genetic background influences severity of immune responses following trauma-hemorrhage.</a> Cytokine. 30, 168-117 (2005).</li><li>Tsukamoto, T., Pape, H. C. <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+for+trauma+research:+What+are+the+options.">Animal models for trauma research: What are the options.</a> Shock. 31, 3-10 (2008).</li><li>DeMaria, E. J., Pellicane, J. V., Lee, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemorrhagic+shock+in+endotoxin+resistant+mice+:+Improved+survival+unrelated+to+deficient+production+of+tumor+necrosis+factor.">Hemorrhagic shock in endotoxin resistant mice : Improved survival unrelated to deficient production of tumor necrosis factor.</a> J Trauma. 35, 720-724 (1993).</li><li>Jamsa, T., Jalovaara, P., Peng, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+three-point+bending+test+and+peripheral+quantitative+computed+tomography+analysis+in+the+evaluation+of+the+strength+of+mouse+femur.">Comparison of three-point bending test and peripheral quantitative computed tomography analysis in the evaluation of the strength of mouse femur.</a> Bone. 23, 155-161 (1998).</li><li>Bounarens, F., Einhorn, 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=Production+of+a+standard+closed+fracture+in+laboratory+animal+bone.">Production of a standard closed fracture in laboratory animal bone.</a> J Ortho Res. 2, 97-101 (1984).</li><li>Holstein, J. H., Menger, M. D., Culemann, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+locking+femur+nail+for+mice.">Development of a locking femur nail for mice.</a> J Biomech. 40, 215-219 (2007).</li><li>Holstein, J. H., Garcia, P., Histing, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+the+establishment+of+defined+mouse+models+for+the+study+of+fracture+healing+and+bone+regeneration.">Advances in the establishment of defined mouse models for the study of fracture healing and bone regeneration.</a> J Orthop Trauma. 23, S31-S38 (2009).</li><li>Histing, T., Garcia, P., Matthys, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+internal+locking+plate+to+study+intramembranous+bone+healing+in+a+mouse+femur+fracture+model.">An internal locking plate to study intramembranous bone healing in a mouse femur fracture model.</a> J Orthop Research. 28, 397-402 .</li><li>Manigrasso, M. B., O'Connor, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+closed+femur+fracture+model+in+mice.">Characterization of a closed femur fracture model in mice.</a> J Ortho Trauma. 18, 687-695 (2004).</li><li>Mark, H., Bergholm, J., Nilsson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+external+fixation+method+and+device+to+study+fracture+healing+in+rats.">An external fixation method and device to study fracture healing in rats.</a> Acta Orthop Scand. 74, 476-487 (2003).</li><li>Bhandari, M., Shanghnessy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+minimally+invasive+percutaneous+technique+of+intramedullary+nail+insertion+in+an+animal+model+of+fracture+healing.">A minimally invasive percutaneous technique of intramedullary nail insertion in an animal model of fracture healing.</a> Arch Orthop Trauma Surg. 121, 591-593 (2001).</li><li>Sonanis, S. V., Lampard, A. L., Kamat, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+technique+to+remove+a+bent+femoral+intramedullary+nail+and+broken+interlocking+screw.">A simple technique to remove a bent femoral intramedullary nail and broken interlocking screw.</a> J Trauma. 63, 435-438 (2007).</li><li>Kobbe, P., Kaczorwoski, D. J., Vodovotz, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+exposure+of+bone+components+to+injured+soft+tissue+induces+Toll-like+receptor+4-dependent+systemic+inflammation+with+acute+lung+injury.">Local exposure of bone components to injured soft tissue induces Toll-like receptor 4-dependent systemic inflammation with acute lung injury.</a> Shock. 30, 686-691 (2008).</li><li>Bumann, M., Henke, T., Gerngross, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+haemorrhagic+shock+on+fracture+healing.">Influence of haemorrhagic shock on fracture healing.</a> Lang Arch Surg. 388, 331-338 (2003).</li><li>Santry, H. P., Alam, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluid+resuscitation:+past,+present,+and+the+future.">Fluid resuscitation: past, present, and the future.</a> Shock. 33, 229-241 (2010).</li><li>Hierholzer, C., Billiar, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+in+the+early+phase+of+hemorrhagic+shock.">Molecular mechanisms in the early phase of hemorrhagic shock.</a> Lang Arch Surg. 386, 302-308 (2001).</li><li>Chaudry, I., Ayala, A., Ertel, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemorrhage+and+resuscitation:+immunological+aspects.">Hemorrhage and resuscitation: immunological aspects.</a> Am J Physiol. 259, 63-678 (1990).</li><li>Kawasaki, T., Hubbard, W. J., Choudhry, 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=Trauma-hemorrhage+induces+depressed+splenic+dendritic+cell+functions+in+mice.">Trauma-hemorrhage induces depressed splenic dendritic cell functions in mice.</a> J Immunol. 177, 4514-4520 (2006).</li><li>Kohut, L., Darwiche, S. S., Frank, A. M., Brumfield, J. M., Billiar, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fixed+volume+or+Fixed+Pressure:+A+Murine+Model+of+Hemorrhgaic+Shock.">Fixed volume or Fixed Pressure: A Murine Model of Hemorrhgaic Shock.</a> J Vis Exp. , (2010).</li><li>Menzel, C. L., Pfeifer, R., Darwiche, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+of+lower+extremity+damage+in+mice:+time+course+of+organ+damage+and+immune+response.">Models of lower extremity damage in mice: time course of organ damage and immune response.</a> J Surg Res. 166, e149-e156 (2011).</li><li>Pfeifer, R., Kobbe, P., Darwiche, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+hemorrhage+in+the+induction+of+systemic+inflammation+and+remote+organ+damage:+Analysis+of+combined+pseudo-fracture+and+hemorrhagic+shock.">Role of hemorrhage in the induction of systemic inflammation and remote organ damage: Analysis of combined pseudo-fracture and hemorrhagic shock.</a> J Orthop Res. 29, 270-274 (2011).</li></ol>

Pseudofracture，重现的无菌肌肉骨骼损伤的小鼠模型，可以评估创伤后的免疫反应。通过休闲的长骨骨折的特点，pseudofracture模型免疫模仿下肢骨折环境：受伤的肌肉和软组织暴露不破坏原生骨骨和骨髓受损 38,39双相免疫反应，可以看出以下。 pseudofracture创伤早期hyperinflammatory的响应，可以看出在6个小时的延迟免疫抑制的第二个组件描述为大约48小时的低谷到高峰组成。这种模式有助于克服?...

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Surgical blue pad</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>50-7105</td>
<td>&nbsp;</td>
<tr>
<td>Sterile Field dressings</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>NC9517505</td>
<td>&nbsp;</td>
<tr>
<td>Circulating heating pad 18"x26"</td>
<td>&nbsp;</td>
<td>Harvard</td>
<td>py872-5272</td>
<td>&nbsp;</td>
<tr>
<td>Hot bead instrument sterilizer</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>11156-002</td>
<td>&nbsp;</td>
<tr>
<td>Stainless steel tray 8" x 11"</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>62687-049</td>
<td>&nbsp;</td>
<tr>
<td>Plexiglass boards (10x15x0.5cm) </td>
<td>&nbsp;</td>
<td>University of Pittsburgh Machine shop</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Tape rolls 1"</td>
<td>&nbsp;</td>
<td>Corporate Express</td>
<td>MMM26001</td>
<td>&nbsp;</td>
<tr>
<td>50cc conical tube</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Straight side wide mouth jars (used as cap for nose cone)</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>159000-058</td>
<td>&nbsp;</td>
<tr>
<td>Oster A5 clippers w. size 40 blade</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>10749-020</td>
<td>&nbsp;</td>
<tr>
<td>Surgical scissors (straight &#x2013; 12cm)</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14068-12</td>
<td>&nbsp;</td>
<tr>
<td>Hemostats curved -18cm</td>
<td>&nbsp;</td>
<td>Harvard</td>
<td>81331718</td>
<td>&nbsp;</td>
<tr>
<td>Forceps (0.8mm-tip, curved-10cm)</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11050-10</td>
<td>&nbsp;</td>
<tr>
<td>Gauze 4"x4"</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1.5cc microfuge tube</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Ice bucket</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Mortar and Pestle </td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>12-961AA</td>
<td>&nbsp;</td>
<tr>
<td>1cc syringe w/ 25G needle</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>14-826-88</td>
<td>&nbsp;</td>
<tr>
<td>20G needle</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1mL pipetteman</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1mL pipette tips</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Falcon polystyrene 8ml tubes </td>
<td>&nbsp;</td>
<td>VWR</td>
<td>60819-331</td>
<td>&nbsp;</td>
<tr>
<td>Sterilization pouch 3"x8"</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>24008</td>
<td>&nbsp;</td>
<tr>
<td>Sterilization pouch 5"x10"</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>24010</td>
<td>&nbsp;</td>
<tr>
<td>MacConkey II Agar plate</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>221172</td>
<td>&nbsp;</td>
<tr>
<td>Ethyl Alcohol - 200 proof </td>
<td>&nbsp;</td>
<td>Pharmaco-AAPER</td>
<td>&nbsp;</td>
<td>[70%]</td>
</tr>
<tr>
<td>Pentobarbital Sodium (Nembutol Sodium Solution)</td>
<td>&nbsp;</td>
<td>Ovation </td>
<td>&nbsp;</td>
<td>70mg/kg</td>
</tr>
<tr>
<td>Aerrane (Isoflurane)</td>
<td>&nbsp;</td>
<td>Baxter </td>
<td>&nbsp;</td>
<td>99.9%</td>
</tr>
<tr>
<td>Triadine Povidone Iodine (Betadine)</td>
<td>&nbsp;</td>
<td>Triad disposables</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Phosphate Buffered Saline (PBS)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Buprenorphine HCl</td>
<td>&nbsp;</td>
<td>Bedford Laboratories</td>
<td>&nbsp;</td>
<td>0.1mg/kg</td>
</tr>		</tbody>
		</table>
		</div>

pseudofracture: an acute peripheral tissue trauma model

创伤后有一个早期的超反应性的炎症反应，可导致多器官功能障碍和创伤患者的死亡率高;这种反应往往伴随着延迟免疫抑制，增加感染的临床并发症， 还可以增加死亡率1-9。许多研究已开始在免疫系统的反应，以评估这些变化的创伤。 10-15 免疫学研究的大力支持，通过多种转基因和基因敲除小鼠体内建模;这些菌株在详细调查援助，以评估参与免疫反应的分子途径 16-21 在实验小鼠创伤建模的挑战是长期的调查，在小鼠骨折的内固定技术，可以复杂，不容易重现 。22-30 这pseudofracture模式，一个很容易复制的创伤模型，克服了这些困难，免疫模仿下肢骨折的环境，同时允许在动物和长期生存的自由运动，没有持续的，长期使用麻醉。其目的是重新长骨骨折的特点，受伤的肌肉和软组织暴露不破坏原生骨受损的骨骼和骨髓。 pseudofracture模型由两部分组成：一个双边的后肢肌肉挤压伤，其次是注射到这些受伤的肌肉骨解决方案，。准备收获从一个年龄和体重相匹配的同源捐助两个后肢长骨骨的解决方案。然后，这些骨头粉碎，重悬于磷酸盐缓冲生理盐水创建骨解决方案。 双侧股骨骨折是四肢创伤中常用的和行之有效的模式，并pseudofracture模型的发展过程中比较模型。各种可用的骨折模型当中，我们选择使用我们的pseudofracture比较软组织损伤的骨折闭合方法，因为我们想无菌但周围组织的比例严重创伤模型。 31 失血性休克是一种严重创伤的设置共同发现，和全球的低灌注创伤模型增加了一个非常相关的元素。32-36的pseudofracture模型可以很容易地与失血性休克模型为多发性创伤的严重程度高的模型相结合37。

Watch this Scientific Journal Video about Pseudofracture: An Acute Peripheral Tissue Trauma Model at JoVE.com

Pseudofracture: An Acute Peripheral Tissue Trauma Model

Pseudofracture，重现的无菌肌肉骨骼损伤的小鼠模型，可以为后期的长期创伤后的免疫反应的评价。本文介绍的模式一步一步的程序执行，包括允许多发性创伤的研究实验模型组合的潜力。

Pseudofracture：急性周围组织的创伤模型

Following trauma there is an early hyper-reactive inflammatory response that can lead to multiple organ dysfunction and high mortality in trauma patients; ...

pseudofracture-an-acute-peripheral-tissue-trauma-model

Research

JoVE Journal

Medicine

Expired CO2 Measurement in Intubated or Spontaneously Breathing Patients from the Emergency Department

Carbon dioxide (CO2) along with oxygen (O2) share the role of being the most important gases in the human body. The measuring of expired CO2 at the mouth has solicited growing clinical interest among physicians in the emergency department for various indications: (1) surveillance et monitoring of the intubated patient; (2) verification of the correct positioning of an endotracheal tube; (3) monitoring of a patient in cardiac arrest; (4) achieving normocapnia in intubated head trauma patients; (5) monitoring ventilation during procedural sedation. The video allows physicians to familiarize themselves with the use of capnography and the text offers a review of the theory and principals involved. In particular, the importance of CO2 for the organism, the relevance of measuring expired CO2, the differences between arterial and expired CO2, the material used in capnography with their artifacts and traps, will be reviewed. Since the main reluctance in the use of expired CO2 measurement is due to lack of correct knowledge concerning the physiopathology of CO2 by the physician, we hope that this explanation and the video sequences accompanying will help resolve this limitation.

Expired CO2 Measurement in Intubated or Spontaneously Breathing Patients from the Emergency Department

This video describes how to perform CO2 measurement in intubated as well as spontaneously breathing patients. The main clinical indications refer to emergency situations: (1) verifying adequate positioning of an endotracheal tube; (2) achieving normocapnia in trauma patients; (3) monitoring ventilation in the case of procedural sedation.

过期的CO 2测量气管插管或自主呼吸的病人从急诊科

Carbon dioxide (CO2) along with oxygen (O2) share the role of being the most important gases in the human body. The measuring of expired CO2 at the mouth ...

科研

医学

Visualization and Analysis of Blood Flow and Oxygen Consumption in Hepatic Microcirculation: Application to an Acute Hepatitis Model

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of the hepatic blood supply is poorly oxygenated portal vein blood derived from the gastrointestinal tract and spleen. Oxygen is delivered to hepatocytes by blood flowing from a terminal branch of the portal vein to a central venule via sinusoids, and this makes an oxygen gradient in hepatic lobules. The oxygen gradient is an important physical parameter that involves the expression of enzymes upstream and downstream in hepatic microcirculation, but the lack of techniques for measuring oxygen consumption in the hepatic microcirculation has delayed the elucidation of mechanisms relating to oxygen metabolism in liver. We therefore used FITC-labeled erythrocytes to visualize the hepatic microcirculation and used laser-assisted phosphorimetry to measure the partial pressure of oxygen in the microvessels there. Noncontact and continuous optical measurement can quantify blood flow velocities, vessel diameters, and oxygen gradients related to oxygen consumption in the liver. In an acute hepatitis model we made by administering acetaminophen to mice we observed increased oxygen pressure in both portal and central venules but a decreased oxygen gradient in the sinusoids, indicating that hepatocyte necrosis in the pericentral zone could shift the oxygen pressure up and affect enzyme expression in the periportal zone. In conclusion, our optical methods for measuring hepatic hemodynamics and oxygen consumption can reveal mechanisms related to hepatic disease.

An optical system was developed to visualize hepatic microcirculation with FITC-labeled erythrocytes and to measure the partial pressure of oxygen in the microvessels with laser-assisted phosphorimetry. This method can be used to investigate physiological and pathological mechanisms by analyzing microvascular structure, diameter, blood flow velocity, and oxygen tension.

可视化和肝脏微循环血流量和耗氧量分析：急性肝炎模型中的应用

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

采用实时定量PCR检测供体细胞植入一个具有竞争力的小鼠骨髓移植模型

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no effective treatment for it yet. Limitations of currently used animal models impede understanding of the pathology mechanism. Multiphoton microscopy allows studying cells and tissues within intact animal brains longitudinally under physiological and pathological conditions. Here, we describe two models of acute brain injury studied by means of two-photon imaging of brain cell behavior under posttraumatic conditions. A selected brain region is injured with a sharp needle to produce a trauma of a controlled width and depth in the brain parenchyma. Our method uses stereotaxic prick with a syringe needle, which can be combined with simultaneous drug application. We propose that this method can be used as an advanced tool to study cellular mechanisms of pathophysiological consequences of acute trauma in mammalian brain in vivo. In this video, we combine acute brain injury with two preparations: cranial window and skull thinning. We also discuss advantages and limitations of both preparations for multisession imaging of brain regeneration after trauma.

Acute brain trauma is a severe injury that has no adequate treatment to date. Multiphoton microscopy allows studying longitudinally the process of acute brain trauma development and probing therapeutical strategies in rodents. Two models of acute brain trauma studied with in vivo two-photon imaging of brain are demonstrated in this protocol.

急性脑外伤小鼠其次是纵向双光子成像

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.

The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses.

在背皮褶商会鼠标的缺血性组织损伤：一个皮瓣模型，探讨急性缺血持续

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are ...

Direct Mouse Trauma/Burn Model of Heterotopic Ossification

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical procedures. The majority of animal models used to study HO rely on the application of exogenous substances, such as bone morphogenetic protein (BMP), exogenous cell constructs, or genetic mutations in BMP signaling. While these models are useful they do not accurately reproduce the inflammatory states that cause the majority of cases of HO. Here we describe a burn/tenotomy model in mice that reliably produces focused HO. This protocol involves creating a 30% total body surface area partial thickness contact burn on the dorsal skin as well as division of the Achilles tendon at its midpoint. Relying solely on traumatic injury to induce HO at a predictable location allows for time-course study of endochondral heterotopic bone formation from intrinsic physiologic processes and environment only. This method could prove instrumental in understanding the inflammatory and osteogenic pathways involved in trauma-induced HO. Furthermore, because HO develops in a predictable location and time-course in this model, it allows for research to improve early imaging strategies and treatment modalities to prevent HO formation.

An Achilles tenotomy and burn injury model of heterotopic ossification allows for the reliable study of trauma induced ectopic bone formation without the application of exogenous factors.

异位骨化直接鼠标创伤/烧伤模型

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical ...

An Acute Retinal Model for Evaluating Blood Retinal Barrier Breach and Potential Drugs for Treatment

A low-cost, easy-to-use and powerful model system is established to evaluate potential treatments that could ameliorate blood retinal barrier breach. An inflammatory factor, histamine, is demonstrated to compromise vessel integrity in the cultured retina through positive staining of IgG outside of the blood vessels. The effects of histamine itself and those of candidate drugs for potential treatments, such as lipoxin A4, are assessed using three parameters: blood vessel leakage via IgG immunostaining, activation of M&#252;ller cells via GFAP staining and change in neuronal dendrites through staining for MAP2. Furthermore, the layered organization of the retina allows a detailed analysis of the processes of M&#252;ller and ganglion cells, such as changes in width and continuity. While the data presented is with swine retinal culture, the system is applicable to multiple species. Thus, the model provides a reliable tool to investigate the early effects of compromised retinal vessel integrity on different cell types and also to evaluate potential drug candidates for treatment.

A low-cost, easy-to-use and powerful system is established to evaluate potential treatments that could ameliorate blood retinal barrier breach induced by histamine. Blood vessel leakage, M&#252;ller cell activation and the continuity of neuronal processes are utilized to assess the damage response and its reversal with a potential drug, lipoxin A4.

急性视网膜模型评估血视网膜屏障违约和潜在的药物治疗

A low-cost, easy-to-use and powerful model system is established to evaluate potential treatments that could ameliorate blood retinal barrier breach. An ...

A Saline/Bipolar Radiofrequency Energy Device As an Adjunct for Hemostasis in Solid Organ Injury/Trauma

Solid organ (liver, spleen, and kidney) hemorrhage is often life-threatening and can be difficult to stop in critically ill patients. Traditional techniques for arresting this ongoing bleeding include coagulation by high voltage electrocautery, topical hemostatic application, and the delivery of ignited argon gas. The goal of this study/video was to demonstrate the efficacy of a new energy device for arresting persistent solid organ hemorrhage.A novel instrument utilizing bipolar radiofrequency (RF) energy which acts to ignite/boil dripping saline from a simple handpiece is employed to arrest ongoing bleeding from solid organ injuries in a porcine model. This instrument is extrapolated from experience within elective hepatic resections. An escalating series of injuries to solid organs within a porcine model will be created. This will be followed by arresting hemorrhage with this novel energy device in sequence. A standard suction device will also be employed. This simple saline/RF energy instrument has the potential to arrest ongoing solid organ surface/capsular bleeding, as well as moderate hemorrhage associated with deep lacerations.

The goal of this publication is to demonstrate the potential application of a novel device using simulated solid organ injuries in a porcine model.

盐水/双极射频能量装置作为固体器官损伤/创伤中血侧体

Solid organ (liver, spleen, and kidney) hemorrhage is often life-threatening and can be difficult to stop in critically ill patients. Traditional ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

An Ex Vivo Tissue Culture Model for Fibrovascular Complications in Proliferative Diabetic Retinopathy

Diabetic retinopathy (DR) is the most common microvascular complication of diabetes and one of the leading causes of blindness in working-age adults. No current animal models of diabetes and oxygen-induced retinopathy develop the full-range progressive changes manifested in human proliferative diabetic retinopathy (PDR). Therefore, understanding of the disease pathogenesis and pathophysiology has relied largely on the use of histological sections and vitreous samples in approaches that only provide steady-state information on the involved pathogenic factors. Increasing evidence indicates that dynamic cell-cell and cell-extracellular matrix (ECM) interactions in the context of three-dimensional (3D) microenvironments are essential for the mechanistic and functional studies towards the development of new treatment strategies. Therefore, we hypothesized that the pathological fibrovascular tissue surgically excised from eyes with PDR could be utilized to reliably unravel the cellular and molecular mechanisms of this devastating disease and to test the potential for novel clinical interventions. Towards this end, we developed a novel method for 3D ex vivo culture of surgically-excised patient-derived fibrovascular tissue (FT), which will serve as a relevant model of human PDR pathophysiology. The FTs are dissected into explants and embedded in fibrin matrix for ex vivo culture and 3D characterization. Whole-mount immunofluorescence of the native FTs and end-point cultures allows thorough investigation of tissue composition and multicellular processes, highlighting the importance of 3D tissue-level characterization for uncovering relevant features of PDR pathophysiology. This model will allow the simultaneous assessment of molecular mechanisms, cellular/tissue processes and treatment responses in the complex context of dynamic biochemical and physical interactions within the PDR tissue architecture and microenvironment. Since this model recapitulates PDR pathophysiology, it will also be amenable for testing or developing new treatments.

Here, we present a protocol to study the pathophysiology of proliferative diabetic retinopathy by using patient-derived, surgically-excised, fibrovascular tissues for three-dimensional native tissue characterization and ex vivo culture. This ex vivo culture model is also amenable for testing or developing new treatments.

增殖性糖尿病视网膜病变纤维血管并发症的体外组织培养模型

Diabetic retinopathy (DR) is the most common microvascular complication of diabetes and one of the leading causes of blindness in working-age adults. No ...