Dexamethasone prevents vascular damage in earlystage non-freezing cold injury of the sciatic nerve

Non-freezing cold injury is a prevalent cause of peripheral nerve damage, but its pathogenic mechanism is poorly understood, and treatment remains inadequate. Glucocorticoids have anti-inflammatory and lipid peroxidation-inhibiting properties. We therefore examined whether dexamethasone, a synthetic glucocorticoid compound, would alleviate early-stage non-freezing cold injury of the sciatic nerve. We established Wistar rat models of non-freezing cold injury by exposing the left sciatic nerve to cold（3–5°C） for 2 hours, then administered dexamethasone（3 mg/kg intraperitoneally） to half of the models. One day after injury, the concentration of Evans blue tracer in the injured sciatic nerve of rats that received dexamethasone was notably lower than that in the injured sciatic nerve of rats that did not receive dexamethasone; neither Evans blue dye nor capillary stenosis was observed in the endoneurium, but myelinated nerve fibers were markedly degenerated in the injured sciatic nerve of animals that received dexamethasone. After dexamethasone administration, however, endoneurial vasculopathy was markedly improved, although damage to the myelinated nerve fiber was not alleviated. These findings suggest that dexamethasone protects the blood-nerve barrier, but its benefit in non-freezing cold injury is limited to the vascular system.

Expert consensus on the prevention, diagnosis and treatment of cold injury in China, 2020

Cold injury refers to local or systemic injury caused by a rapid, massive loss of body heat in a cold environment. The incidence of cold injury is high. However, the current situation regarding the diagnosis and treatment of cold injury in our country is not ideal. To standardize and improve the level of clinical diagnosis and treatment of cold injury in China, it is necessary to make a consensus that is practical and adapted to the conditions in China. We used the latest population-level epidemiological and clinical research data, combined with relevant literature from China and foreign countries. The consensus was developed by a joint committee of multidisciplinary experts. This expert consensus addresses the epidemiology, diagnosis, on-site emergency procedures, in-hospital treatment, and prevention of cold injury.

Is it time to put traditional cold therapy in rehabilitation of soft-tissue injuries out to pasture?

Cold therapy has been used regularly as an immediate treatment to induce analgesia following acute soft-tissue injuries,however,a prolonged ice application has proved to delay the start of the healing and lengthen the recovery process.Hyperbaric gaseous cryotherapy,also known as neurocryostimulation,has shown the ability to overcome most of the limitations of traditional cold therapy,and meanwhile promotes the analgesic and anti-inflammatory effects well,but the current existing studies have shown conflicting results on its effects.Traditional cold therapy still has beneficial effect especially when injuries are severe and swelling is the limiting factor for recovery after soft-tissue injuries,and therefore no need to be entirely put out to pasture in the rehabilitation practice.Strong randomized controlled trials with good methodological quality are still needed in the future to evaluate the effects of different cryotherapy modalities.

Cold water swimming pretreatment reduces cognitive deficits in a rat model of traumatic brain injury

A moderate stress such as cold water swimming can raise the tolerance of the body to potentially injurious events. However, little is known about the mechanism of beneficial effects induced by moderate stress. In this study, we used a classic rat model of traumatic brain injury to test the hypothesis that cold water swimming preconditioning improved the recovery of cognitive functions and explored the mechanisms. Results showed that after traumatic brain injury, pre-conditioned rats（cold water swimming for 3 minutes at 4℃） spent a significantly higher percent of times in the goal quadrant of cold water swim, and escape latencies were shorter than for non-pretreated rats. The number of circulating endothelial progenitor cells was significantly higher in pre-conditioned rats than those without pretreatment at 0, 3, 6 and 24 hours after traumatic brain injury. Immunohistochemical staining and Von Willebrand factor staining demonstrated that the number of CD34~＋ stem cells and new blood vessels in the injured hippocampus tissue increased significantly in pre-conditioned rats. These data suggest that pretreatment with cold water swimming could promote the proliferation of endothelial progenitor cells and angiogenesis in the peripheral blood and hippocampus. It also ameliorated cognitive deficits caused by experimental traumatic brain injury.

Warm HTK donor pretreatment reduces liver injury during static cold storage in experimental rat liver transplantation

BACKGROUND： Organ shortage has led to an increased number of transplantations from extended criteria donors. These organs are more vulnerable to ischemia-reperfusion injury. Thus, improvement of organ preservation is needed. HTK is a widely used preservation solution for static cold storage in liver transplantation. The present study was to investigate the beneficial effect of warm HTK donor pretreatment on liver preservation.

Dose-dependent neuroprotective effect of enoxaparin on cold-induced traumatic brain injury

Recent evidence exists that enoxaparin can reduce brain injury because of its anticoagulant activity. To investigate the potential therapeutic effect of enoxaparin on cold-induced traumatic brain injury, at 20 minutes after modeling, male BALB/c mouse models of cold-induced traumatic brain injury were intraperitoneally administered 3 and 10 mg/kg enoxaparin or isotonic saline solution. Twenty-four hours later, enoxaparin at 10 mg/kg greatly reduced infarct volume, decreased cell apoptosis in the cortex and obviously increased serum level of total antioxidant status. By contrast, administration of enoxaparin at 3 mg/kg did not lead to these changes. These findings suggest that enoxaparin exhibits neuroprotective effect on cold-induced traumatic brain injury in a dose-dependent manner.

Neuroprotective effects of cold-inducible RNA-binding protein during mild hypothermia on traumatic brain injury

Cold-inducible RNA-binding protein（CIRP）, a key regulatory protein, could be facilitated by mild hypothermia in the brain, heart and liver. This study observed the effects of mild hypothermia at 31 ± 0.5℃ on traumatic brain injury in rats. Results demonstrated that mild hypothermia suppressed apoptosis in the cortex, hippocampus and hypothalamus, facilitated CIRP m RNA and protein expression in these regions, especially in the hypothalamus. The anti-apoptotic effect of mild hypothermia disappeared after CIRP silencing. There was no correlation between mitogen-activated extracellular signal-regulated kinase activation and CIRP silencing. CIRP silencing inhibited extracellular signal-regulated kinase-1/2 activation. These indicate that CIRP inhibits apoptosis by affecting extracellular signal-regulated kinase-1/2 activation, and exerts a neuroprotective effect during mild hypothermia for traumatic brain injury.

Analysis on Meteorological Conditions of Winter Cold Injury of Sugar Orange Industry in Guilin

Yield and quality of sugar orange are closely related to meteorological conditions,especially the low temperature and cold injury in mature period of fruit. To study the influence of winter cold injury on sugar orange industry of Guilin,the chilling injury weather in each county of Guilin planting sugar orange is analyzed,and the disastrous cold weather and its defense measures are studied. Frost,freezing and cold wave happen in Guilin winter sometimes,and the disastrous weather could seriously harm maturing sugar orange or that in the tree,causing peel coke,dehiscent fruit,fruit drop and quality decline. It can effectively reduce the loss by chilling injury and guarantee excellent quantity and high yield of sugar orange by timely understanding weather change information,scientifically managing water and fertilizer,and doing film cover well.

Corona virus disease 2019-associated liver injury in cold regions

The corona virus disease 2019(COVID-19)pandemic has created a global health and economic crisis.Our studies uncovered that in addition to respiratory symptoms,liver damage is also common in COVID-19 patients;however,the cause of liver damage has not been fully elucidated.In this article,we summarize the clinical manifestations and pathological features of COVID-19 reported in published relevant studies and delineate the etiology and pathogenesis of COVID-19-related liver injury.We speculate that cold stimulation may be associated with COVID-19-related liver injury,which should be considered in clinical decision-making and treatment of COVID-19 in cold regions.

BH3-only protein Bim is involved in myocardial injury induced by co-stress of ischemia and cold stress in rats

Objectives To investigate the effect of co-exposure of myocardial ischemia and cold stress on myocardial injury in rats and the relative mechanism.Methods Myocardial ischemia model was established by ligation of left coronary artery.SD rats were randomly allocated to 4 groups; sham+normal temperature(S group),sham+cold stress(SC group),myocardial ischemia+ normal temperature(Ⅰgroup), myocardial ischemia+cold stress(IC group).On the condition of 26℃,SC and IC groups were keeped in a 4℃artificial chamber for 8h(8;00-16:00) for 4 consecu- tive days.Car diac function was assessed by echocardiography;pathological change was analyzed by HE staining;myocardial infarct size was determined by TTC staining;Bim,Caspase-3 expression in myocardium was determined by western blotting.Results It was demonstrated that co-exposure of myocardial ischemia and cold stress could significantly make the cardiac muscle in abnormal shape,increase the infarct size and the expression of Bim and Caspase-3.Conclusions Co-exposure of myocardial ischemia and cold stress may aggravate the cardiac injury,pro- apoptosis protein Bim is involved.

Research progress on combat trauma treatment in cold regions

Cold regions are a special combat environment in which low temperatures have a great impact on human metabolism and other vital functions, including the nervous, motion, cardiovascular, circulatory, respiratory, and urinary systems; consequently, low temperatures often aggravate existing trauma, leading to high mortality rates if rapid and appropriate treatment is not provided. Hypothermia is an independent risk factor of fatality following combat trauma; therefore, proactive preventative measures are needed to reduce the rate of mortality. After summarizing the basic research on battlefield environments and progress in the prevention and treatment of trauma, this article concludes that current treatment and prevention measures for combat trauma in cold regions are inadequate. Future molecular biology studies are needed to elucidate the mechanisms and relevant cell factors underlying bodily injury caused by cold environment, a research goal will also allow further exploration of corresponding treatments.

Liver graft preservation methods during cold ischemia phase and normothermic machine perfusion

The growing demand for donor organs requires measures to expand donor pool.Those include extended criteria donors, such as elderly people, steatotic livers,donation after cardiac death, etc. Static cold storage to reduce metabolic requirements developed by Collins in late 1960 s is the mainstay and the golden standard for donated organ protection. Hypothermic machine perfusion provides dynamic organ preservation at 4°C with protracted infusion of metabolic substrates to the graft during the ex vivo period. It has been used instead of static cold storage or after it as short perfusion in transplant center. Normothermic machine perfusion(NMP) delivers oxygen, and nutrition at physiological temperature mimicking regular environment in order to support cellular function. This would minimize effects of ischemia/reperfusion injury.Potentially, NMP may help to estimate graft functionality before implantation into a recipient. Clinical studies demonstrated at least its non-inferiority or better outcomes vs static cold storage. Regular grafts donated after brain death could be safely preserved with convenient static cold storage. Except for prolonged ischemia time where hypothermic machine perfusion started in transplant center could be estimated to provide possible positive reconditioning effect. Use of hypothermic machine perfusion in regular donation instead of static cold storage or in extended criteria donors requires further investigation. Multicenter randomized clinical trial supposed to be completed in December 2021. Extended criteria donors need additional measures for graft storage and assessment until its implantation. NMP is actively evaluating promising method for this purpose.Future studies are necessary for precise estimation and confirmation to issue clinical practice recommendations.

Melatonin treatment alleviates chilling injury in mango fruit 'Keitt' by modulating proline metabolism under chilling stress

Mangoes often suffer from low temperature-induced chilling injury (CI) during postharvest cold storage.Therefore,advanced techniques are crucial and in high demand to solve the chilling stress of mango fruit for a higher value.This study addresses chilling stress modulation by investigating the effects of melatonin treatment on CI,proline metabolism,and related gene expressions of ‘Keitt’ mango during cold storage after dipped in 0 (control),0.1 (MT1),and 0.2 mmol L^(-1)(MT2) melatonin solution for 30 min.The results revealed that melatonin treatment in MT1 significantly reduced CI development and increased proline content in mango fruit during cold storage compared to the control.These changes were along with increases in the activity of critical enzymes as well as the expression of encoding genes involved in proline biosynthesis,such as pyrroline-5-carboxylate synthetase (P5CS),pyrroline-5-carboxylate reductase (P5CR),ornithine D-aminotransferase (OAT),P5CS2,P5CR2,and OAT3.Additionally,proline dehydrogenase (PDH) activity and the expression of the PDH3 gene associated with proline dehydrogenation were lower in MT1-treated mangoes than the controlled group.Thus,melatonin treatment has regulated proline metabolism resulting in the accumulation of proline,subsequently contributing to enhancing the chilling tolerance of‘Keitt’mango fruit.

Gene expression profile in rat small intestinal allografts after cold preservation/reperfusion

AIM: To determine the changes of gene expression profile in small intestinal allografts in rats after cold preservation/ reperfusion, and to identify the genes relevant to cold preservation/reperfusion injury. METHODS: Heterotopic segmental small bowel transplantation was performed in six rats with a sham operation and they were used as controls. Total RNA was extracted from the allografts (experimental group) and normal intestines (control group) 1 h after cold preservation/ reperfusion, and then purified to mRNA, which was then reversely transcribed to cDNA, and labeled with fluorescent Cy5-dUTP and Cy3-dUTP to prepare hybridization probes. The mixed probes were hybridized to the cDNA microarray. After high-stringent washing, the fluorescent signals on cDNA microarray chip were scanned and analyzed. RESULTS: Among the 4 096 target genes, 82 differentially expressed genes were identified between the two groups. There were 18 novel genes, 33 expression sequence tags, and 31 previously reported genes. The selected genes may be divided into four classes: genes modulating cellular adhesion, genes regulating cellular energy, glucose and protein metabolism, early response genes and other genes. CONCLUSION: A total of 82 genes that may be relevant to cold preservation/reperfusion injury in small intestinal allografts are identified. Abnormal adhesion between polymorphonuclears and endothelia and failure in energy, glucose and protein metabolism of the grafts may contribute to preservation/reperfusion injury. The functions of the novel genes identified in our study need to be clarified further.

The Primary Motor Cortex Stimulation Attenuates Cold Allodynia in a Chronic Peripheral Neuropathic Pain Condition in <i>Rattus norvegicus</i>

Background: The primary motor cortex (M1) stimulation (MCS) is a useful tool for attenuation of the peripheral neuropathic pain in patients with pharmacologically refractory pain. Furthermore, that neurological procedure may also cause antinociception in rodents with neuropathic pain. Cold allodynia is a frequent clinical finding in patients with neuropathic pain, then, we evaluated if an adapted model of neuropathy induced by chronic constriction injury (CCI) of the ischiadicus nervus (sciatic nerve) produces cold allodynia in an animal model of chronic pain. In addition, we also investigated the effect of the electrical stimulation of the M1 on chronic neuropathic pain condition in laboratory animals. Methods: Male Wistar rats were used. An adapted model of peripheral mononeuropathy induced by CCI was carried out by placing a single loose ligature around the right sciatic nerve. The acetone test was used to evaluate the cold allodynia in CCI or Sham (without ligature) rats. The MCS (M1) was performed at low-frequency (20 μA, 100 Hz) during 15 s by deep brain stimulation (DBS-Thomas Recording device) 21 days after CCI or Sham procedures. The cold allodynia was measured before and immediately after the neurostimulation of M1 in the following time-window: 0, 15 and 30 min after MCS. Results: Cold allodynia threshold increased in animals with chronic neuropathic pain submitted to the acetone test 21 days after the CCI surgery. The M1-stimulation by DBS procedure decreased the cold allodynia immediately and until 30 min after M1-stimulation in rats with chronic neuropathic pain. Conclusion: The current proposal for a CCI model by a single loose ligature of the sciatic nerve can be employed as an experimental model of chronic neuropathic pain in rats submitted to peripheral nervous system injury. The M1-stimulation produced antinociception in rats with chronic neuropathic pain. Thus, we reinforced that the MCS decreases cold allodynia in laboratory animals submitted to persistent sciatic nerve constriction and can be a more reasonable procedure for the treatment of chronic intractable neuropathic pain.

冷休克蛋白RBM3在幼鼠肾脏缺血再灌注损伤中的作用研究

目的建立幼鼠肾脏缺血再灌注损伤(RIRI)模型,探讨亚低温环境诱导的冷休克蛋白RBM3对RIRI的作用。方法将7 d SD幼鼠24只随机分为正常对照组(NC组)、假手术组(Sham组)、RIRI未亚低温组(NH组)、RIRI亚低温组(TH组),每组6只。建立肾脏RIRI模型,进行全身亚低温治疗;检测血肌酐(SCr)、半胱氨酸蛋白酶抑制剂C(CysC)水平;取左肾组织行HE染色,观察病理学变化,取右肾组织用qPCR检测YAP1和NRF2基因表达情况;用Western Blot检测RBM3蛋白表达量。结果NC组与Sham组的SCr、CysC水平比较,差异无统计学意义(P>0.05);NH组与TH组的SCr、CysC水平显著高于NC组,NH组的SCr、CysC水平显著高于TH组(P<0.05)。NH组和TH组的病理组织评分高于NC组,NH组的病理组织评分高于TH组(P<0.05)。TH组的YAP1、NRF2基因表达显著高于NC组和NH组(P<0.05)。TH组的RBM3蛋白表达量明显高于NC组和NH组(P<0.05)。结论全身亚低温诱导的RBM3对幼鼠RIRI有一定的保护作用,这种保护作用可能与低温诱导的RBM3降低了炎症反应有关。

多参数功能MRI评价冷缺血再灌注肾损伤大鼠肾脏血流及氧合水平的价值

目的探讨动脉自旋标记(ASL)和血氧水平依赖(BOLD)技术评估冷缺血再灌注损伤(CIRI)大鼠肾脏血流和血氧微观改变的价值。方法将100只健康雄性SD大鼠随机分为4组,分别为肾脏冷缺血1 h组(CIRI 1 h)、冷缺血2 h组(CIRI 2 h)、冷缺血4 h组(CIRI 4 h)组及假手术组(CIRI 0 h),每组25只。每组随机选取5只分别在术前、术后(1 h、1 d、2 d、5 d)进行ASL及BOLD MRI,测量肾皮质的血流量(RBFCo)值及T_(2)^(*)值(T_(2)^(*)Co)、肾外的髓外带T_(2)^(*)值(T_(2)^(*)OSOM)和肾外髓内带T^(2*)值(T^(2*)ISOM)。随后切除大鼠左肾进行病理学分析并对肾小管损伤程度评分。采用单因素方差分析比较各组间及组内的不同时间点之间MRI参数和大鼠肾小管损伤程度评分之间的差异;采用Pearson相关分析MRI参数间及MRI参数与肾小管损伤程度评分的相关性。结果术后1 h,冷缺血组大鼠肾脏RBFCo、T_(2)^(*)_(Co)、T_(2)^(*)_(OSOM)值均较术前减低(均P<0.05);术后5 d,CIRI 0 h、CIRI 1 h组的RBFCo、T_(2)^(*)_(Co)、T_(2)^(*)_(OSOM)、T^(2*)_(ISOM)值恢复至其术前水平(均P>0.05),CIRI 4 h组上述指标仍低于术前水平(均P<0.05)。术后1 h、1 d、2 d,CIRI 4 h组T_(2)^(*)OSOM值均较其他各组低;术后5 d,CIRI 4 h组RBFCo、T^(2*)_(Co)、T^(2*)_(OSOM)、T^(2*)_(ISOM)值均低于CIRI 0 h组(均P<0.05)。CIRI 2 h组、CIRI 4 h组的术后各时间点的肾小管损伤程度评分均高于其术前(均P<0.05),术后随时间延长,评分呈减低趋势;术后各时间点,CIRI 4 h组评分均较CIRI 0 h、CIRI 1 h组高;术后1 h、2 d、5 d,CIRI 2 h组的评分均较CIRI 0 h、CIRI 1 h组的要高(均P<0.05)。RBF_(Co)、T_(2)^(*)Co、T_(2)^(*)_(OSOM)、T^(2*)_(ISOM)值与肾小管损伤评分均呈中度负相关(r=-0.714、-0.689、-0.518、-0.579,均P<0.001);RBFCo值与T_(2)^(*)_(Co)、T^(2*)_(OSOM)、T^(2*)_(ISOM)值呈中度正相关(r=0.704、0.525、0.612,均P<0.001);T_(2)^(*)_(Co)值与T_(2)^(*)_(OSOM)、T_(2)^(*)_(ISOM)值呈中度正相关(r=0.685、0.572,均P<0.001);T_(2)^(*)_(OSOM)值与T_(2)^(*)_(ISOM)值呈中度正相关(r=0.606,P<0.001)。结论ASL和BOLD成像可动态评估肾CIRI后组织血流及氧合水平的微观变化,为早期发现肾CIRI提供影像学依据。

冷水浸泡剂量对骨骼肌运动性疲劳恢复效果的系统评价与Meta分析

目的:冷水浸泡恢复方法在浸泡水温、时长及深度等操作性指标上暂无统一方案,导致在骨骼肌运动性疲劳恢复疗效方面产生争议。文章以此分析不同因素下冷水浸泡对肌肉损伤、肌肉酸痛及肌力恢复的影响,以期探寻最佳的浸泡实施方案,从而为肌肉组织疲劳恢复提供证据。方法:对中国知网、万方数据、Web of Science、PubMed数据库进行检索,检索跨度为2000-01-01/2023-08-15,共得文献4759篇,通过筛选排除4735篇,最终纳入文献24篇。采用物理治疗证据数据库量表对纳入文献进行方法学质量评估,并使用Stata-MP 16软件进行效应量合并、亚组分析、Meta回归、敏感性检验和发表偏倚分析。结果:(1)共纳入24项随机对照试验,包括617例受试者,整体上方法学质量较高;(2)Meta分析结果显示:冷水浸泡能显著降低肌酸激酶血值(SMD=-0.17,95%CI:-0.29至-0.05,P<0.01),减轻肌肉酸痛(SMD=-0.60,95%CI:-0.81至-0.38,P<0.01)和促进最大肌力恢复(SMD=0.17,95%CI:0.05-0.30,P<0.01);(3)亚组分析结果显示,水温>14℃(SMD=-0.48,95%CI:-0.76至-0.20,P<0.01)和时长12-14min(SMD=-0.38,95%CI:-0.61至-0.15,P<0.01)的浸泡方案降低肌酸激酶血值效果最好,且对耐力运动(SMD=-0.45,95%CI:-0.71至-0.20,P<0.01)后的干预疗效更显著;水温<10℃(SMD=-0.61,95%CI:-0.79至-0.43,P<0.01)、时长<12 min(SMD=-0.76,95%CI:-0.98至-0.53,P<0.01)以及髂棘以上(SMD=-0.74,95%CI:-0.97至-0.52,P<0.01)的浸泡深度缓解肌肉酸痛效果最好,且对耐力运动(SMD=-0.42,95%CI:-0.61至-0.22,P<0.01)后的镇痛效果更明显;(4)Meta回归结果显示,浸泡水温、浸泡时长及运动类型是影响肌酸激酶效应量的重要调节因子;浸泡水温及浸泡深度是影响目测类比评分效应量的重要调节因子,运动类型是影响最大等长肌力效应量的重要调节因子。结论:(1)极低到中等强度证据显示,冷水浸泡能有效减轻肌肉损伤、缓解肌肉酸痛和促进肌力恢复;(2)在减轻肌肉损伤方面,浸泡水温、浸泡时长、运动类型是影响浸泡疗效的显著调节因子,其中浸泡水温>14℃、时长12-14 min是降低运动后肌肉损伤的最佳方案,且对耐力运动后的浸泡效果更好;(3)在降低肌肉酸痛方面,浸泡水温、浸泡深度是影响干预效应的重要调节因子,其中浸泡水温<10℃、时长<12 min、髂棘以上浸泡深度是减轻肌肉酸痛的最佳方案,且对耐力运动后的镇痛效果更好;(4)在促进肌力恢复方面,运动类型是影响最大等长肌力效应量的关键调节因子。

冷缺血期一氧化碳和氢气联合膨肺对肺移植后大鼠肺损伤的影响

目的 观察冷缺血期一氧化碳(CO)和氢气(H_(2))联合膨肺对大鼠移植肺脏炎症反应的影响及机制。方法 选择成年SPF级雄性SD大鼠80只,40只供体和40只受体,8~10周龄,体重250~280 g。采用随机数字表法将大鼠分为四组:O_(2)膨肺组(O_(2)组)、CO膨肺组(CO组)、H_(2)膨肺组(H_(2)组)和CO和H_(2)联合膨肺组(CH组),每组10对(每对1只供体和1只受体)。供体大鼠切取左肺后,于冷缺血期进行气体膨肺,O_(2)组采用40%O_(2)+60%N_(2)膨肺,CO组采用0.05%CO+40%O_(2)+59.95%N_(2)膨肺,H_(2)组采用3%H_(2)+40%O_(2)+57%N_(2)膨肺,CH组采用0.05%CO+3%H_(2)+40%O_(2)+56.95%N_(2)膨肺,四组体积均为5 ml/kg,每30分钟使用气密针置换气体1次,180 min后行肺移植。受体大鼠于移植前即刻、再灌注后3、60、120、180 min进行血气分析,记录PaO_(2)/FiO_(2)、pH和碱剩余(BE)。再灌注后180 min处死受体大鼠,测定移植肺组织湿/干重比(W/D),采用ELISA法检测移植肺组织白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)浓度及髓过氧化物酶(MPO)含量。检测移植肺组织支气管肺泡灌洗液(BALF)中性粒细胞百分比,采用高效液相色谱法检测BALF中大聚积体(LA)、小聚积体(SA)和LA/SA。采用Western blot法检测移植肺组织表面活性蛋白A(SP-A)和核转录因子-κB(NF-κB)蛋白含量。采用HE染色法观察移植肺组织病理形态并进行肺组织损伤评分(LIS)。结果 与O_(2)组比较,CO组、H_(2)组和CH组受体大鼠再灌注后60、120、180 min PaO_(2)/FiO_(2)、pH、BE均明显升高(P<0.05),移植肺组织W/D、IL-6、IL-1β、TNF-α浓度、MPO含量、中性粒细胞百分比、SA、NF-κB蛋白含量和LIS均明显降低(P<0.05),IL-10浓度、LA、LA/SA和SP-A蛋白含量明显升高(P<0.05)。与CO组比较,H_(2)组和CH组受体大鼠再灌注后120、180 min PaO_(2)/FiO_(2)、pH、BE均明显升高(P<0.05),CH组移植肺组织W/D、IL-6、IL-1β、TNF-α浓度、MPO含量、中性粒细胞比、SA、NF-κB蛋白含量和LIS明显降低(P<0.05),IL-10浓度、LA、LA/SA和SP-A蛋白含量明显升高(P<0.05)。与H_(2)组比较,CH组受体大鼠再灌注后120、180 min PaO_(2)/FiO_(2)、pH、BE均明显升高(P<0.05),移植肺组织W/D、IL-6、IL-1β、TNF-α浓度、MPO含量、中性粒细胞百分比、SA、NF-κB蛋白含量和LIS均明显降低(P<0.05),IL-10浓度、LA、LA/SA和SP-A蛋白含量明显升高(P<0.05)。结论 冷缺血期CO和H_(2)联合膨肺抑制移植肺脏炎症反应,降低NF-κB蛋白含量、升高SP-A蛋白含量、维持肺泡表面活性物质稳定。

4℃冷藏保存血小板输注对大鼠失血性休克/复苏致急性肾损伤的影响研究

目的研究4℃冷藏保存血小板(cold storage platelets,CSP)对大鼠失血性休克/复苏(HS/R)导致急性肾损伤的影响。方法健康雄性SD大鼠40只,随机选取20只用以制备血小板制剂,剩余大鼠随机分为4组:假手术组(空白组)、HS/R模型组(模型组)、CSP输注组、22℃±2℃振荡保存血小板输注组(RTP输注组)。容量复苏后24 h处死动物,采集血和肾组织标本。检测各组血清中血肌酐(Scr)、尿素氮(BUN)、血清胱抑素C(CysC)及肾组织中白细胞介素1β(IL-1β)、白细胞介素18(IL-18)含量。Western Blot法检测各组大鼠肾组织中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、肾损伤因分子1(KIM-1)表达量,HE染色观察肾组织病理学改变。结果①四组大鼠基线状态体质量、MAP、SCr、BUN等指标无明显差异(P>0.05)。②复苏后24 h,模型组大鼠SCr(P<0.05)、BUN(P<0.01)、CysC(P<0.01)水平均高于其他三组;CSP输注组BUN(P<0.01)、SCr(P=0.037)、CysC(P=0.020)水平低于RTP输注组。③与模型组比较,CSP输注组和RTP输注组血清IL-1β和IL-18水平均显著降低。较于RTP输注组,CSP输注组血清IL-1β水平有所降低(P=0.041)。④与模型组相比,CSP输注组NGAL蛋白表达明显下降(P<0.01),CSP输注组和RTP输注组KIM-1蛋白表达均显著下降(P<0.01)。肾组织病理学结果示,CSP输注可减轻肾细胞变性、坏死及炎性细胞浸润程度。结论较于RTP,CSP输注可减轻SD大鼠HS/R急性肾损伤。