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However, time-out application of tourniquets leads to ischemic-necrosis of skeletal muscles and ischemia-reperfusion injury. Regional hypothermia (RH) on wounded limb can relieve the injury on local tissue and distant organs. We aimed to investigate the protective effects of RH on rabbits\u2019 limbs injured by a steel-ball combined with hemorrhagic-shock, and then employed tourniquet over-time, tried to identify the optimal treatment RH.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Thirty rabbits were randomly divided into 5 groups. All rabbits were anesthetized, intubated femoral artery and vein in right-hind limbs. Sham operation group (<jats:bold>Sham<\/jats:bold>): only femoral arteriovenous cannula in right-hind limb. None RH group (<jats:bold>NRH<\/jats:bold>): rabbits were intubated as Sham group, then the soft tissues of rabbits\u2019 left-hinds were injured by a steel-ball shooting, and were exsanguinated until shock, then bundled with rubber tourniquets for 4\u2009h.<jats:bold>Three RH subgroups<\/jats:bold>: rabbits were injured as mentioned above, the injured limbs were bundled with rubber tourniquets and treated with different temperature (5\u2009\u00b1\u20091\u2009\u00b0C, 10\u2009\u00b1\u20091\u2009\u00b0C, and 20\u2009\u00b1\u20091\u2009\u00b0C, respectively) for 4\u2009h. The injury severity of lung and regional muscle was assessed by histologic examination. Activity of adenosine triphosphatase (ATPase) and content of malondialdehyde (MDA) in muscle, inflammatory cytokines, myoglobin, creatine kinase-MM (CK-MM), Heme, Heme oxygenase 1 (HO-1), lactic acid (Lac), and lectrolyte ion in serum were detected.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Following with RH treatment, the injury of lung and local muscle tissue was alleviated evidencing by mitigation of histopathological changes, significant decrease of water-content and MDA content, and increase of ATPase activity. Lower level of Lac, Potassium (K<jats:sup>+<\/jats:sup>), inflammatory cytokines, Heme, CK-MM, myoglobin content, and higher level of Calcium (Ca<jats:sup>2+<\/jats:sup>), HO-1 content were shown in RH treatment. 10\u2009\u00b0C was the most effective RH to increase ATPase activity, and decrease MDA, myoglobin, CK-MM content.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Transient RH (4\u2009h) had a \u201clong-term mitigation effects\u201d (continued for 6\u2009h) on time-out application of tourniquet with the fluid resuscitation and core temperature maintenance, and the most effective temperature for reducing the side effects on tourniquet time-out application was 10\u2009\u00b0C.<\/jats:p><\/jats:sec>","DOI":"10.1186\/s13049-019-0678-3","type":"journal-article","created":{"date-parts":[[2019,11,21]],"date-time":"2019-11-21T14:03:34Z","timestamp":1574345014000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Regional hypothermia attenuates secondary-injury caused by time-out application of tourniquets following limb fragments injury combined with hemorrhagic shock"],"prefix":"10.1186","volume":"27","author":[{"given":"Changmei","family":"Weng","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Lan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liangchao","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianmin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinan","family":"Lai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,11,21]]},"reference":[{"issue":"9\u201310","key":"678_CR1","first-page":"283","volume":"115","author":"Z Stanec","year":"1993","unstructured":"Stanec Z, Skrbic S, Depina I, Hulina D, Ivrlac R, Unusic J, Montani D, Prpic I, Unusic I. 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