Brachial plexus avulsion (BPA) is a combined injury involving the central and peripheral nervous systems. Patients with BPA often experience severe neuropathic pain (NP) in the affected limb. NP is insensitive to the existing treatments, which makes it a challenge to researchers and clinicians. Accumulated evidence shows that a BPA-induced pain state is often accompanied by sympathetic nervous dysfunction, which suggests that the excitation state of the sympathetic nervous system is correlated with the existence of NP. However, the mechanism of how somatosensory neural crosstalk with the sympathetic nerve at the peripheral level remains unclear. In this study, through using a novel BPA C7 root avulsion mouse model, we found that the expression of BDNF and its receptor TrκB in the DRGs of the BPA mice increased, and the markers of sympathetic nervous system activity including α1 and α2 adrenergic receptors (α1-AR and α2-AR) also increased after BPA. The phenomenon of superexcitation of the sympathetic nervous system, including hypothermia and edema of the affected extremity, was also observed in BPA mice by using CatWalk gait analysis, an infrared thermometer, and an edema evaluation. Genetic knockdown of BDNF in DRGs not only reversed the mechanical allodynia but also alleviated the hypothermia and edema of the affected extremity in BPA mice. Further, intraperitoneal injection of adrenergic receptor inhibitors decreased neuronal excitability in patch clamp recording and reversed the mechanical allodynia of BPA mice. In another branch experiment, we also found the elevated expression of BDNF, TrκB, TH, α1-AR, and α2-AR in DRG tissues from BPA patients compared with normal human DRGs through western blot and immunohistochemistry. Our results revealed that peripheral BDNF is a key molecule in the regulation of somatosensory-sympathetic coupling in BPA-induced NP. This study also opens a novel analgesic target (BDNF) in the treatment of this pain with fewer complications, which has great potential for clinical transformation.
Humans ; Mice ; Animals ; Hyperalgesia/metabolism* ; Brain-Derived Neurotrophic Factor/metabolism* ; Hypothermia/metabolism* ; Neuralgia ; Brachial Plexus/injuries* ; Edema/metabolism*

BACKGROUNDAntegrade selective cerebral perfusion (ASCP) is regarded to perform cerebral protection during the thoracic aorta surgery as an adjunctive technique to deep hypothermic circulatory arrest (DHCA). However, brain metabolism profile after ASCP has not been systematically investigated by metabolomics technology.

METHODSTo clarify the metabolomics profiling of ASCP, 12 New Zealand white rabbits were randomly assigned into 60 min DHCA with (DHCA+ASCP [DA] group, n = 6) and without ( DHCA [D] group, n = 6) ASCP according to the random number table. ASCP was conducted by cannulation on the right subclavian artery and cross-clamping of the innominate artery. Rabbits were sacrificed 60 min after weaning off cardiopulmonary bypass. The metabolic features of the cerebral cortex were analyzed by a nontargeted metabolic profiling strategy based on gas chromatography-mass spectrometry. Variable importance projection values exceeding 1.0 were selected as potentially changed metabolites, and then Student's t-test was applied to test for statistical significance between the two groups.

RESULTSMetabolic profiling of brain was distinctive significantly between the two groups (Q 2 Y = 0.88 for partial least squares-DA model). In comparing to group D, 62 definable metabolites were varied significantly after ASCP, which were mainly related to amino acid metabolism, carbohydrate metabolism, and lipid metabolism. Kyoto Encyclopedia of Genes and Genomes analysis revealed that metabolic pathways after DHCA with ASCP were mainly involved in the activated glycolytic pathway, subdued anaerobic metabolism, and oxidative stress. In addition, L-kynurenine (P = 0.0019), 5-methoxyindole-3-acetic acid (P = 0.0499), and 5-hydroxyindole-3-acetic acid (P = 0.0495) in tryptophan metabolism pathways were decreased, and citrulline (P = 0.0158) in urea cycle was increased in group DA comparing to group D.

CONCLUSIONSThe present study applied metabolomics analysis to identify the cerebral metabolic profiling in rabbits with ASCP, and the results may shed new lights that cerebral metabolism is better preserved by ASCP compared with DHCA alone.

Animals ; Brain ; metabolism ; Cerebrovascular Circulation ; Circulatory Arrest, Deep Hypothermia Induced ; Humans ; Male ; Metabolomics ; Rabbits

Hypoxic-ischemic encephalopathy (HIE) in neonates is the brain injury caused by perinatal asphyxia or hypoxia and is a major cause of death in neonates and nervous system dysfunction in infants and young children. Although to a certain degree, mild hypothermia therapy reduces the mortality of infants with moderate to severe HIE, it cannot achieve the expected improvements in nervous system dysfunction. Hence, it is of vital importance to search for effective therapeutic methods for HIE. The search for more therapies and better preventive measures based on the pathogenesis of HIE has resulted in much research. As an important link in the course of HIE, energy failure greatly affects the development and progression of HIE. This article reviews the research advances in the treatment and prevention of energy failure in the course of HIE.
Energy Metabolism ; Humans ; Hypothermia, Induced ; Hypoxia-Ischemia, Brain ; prevention & control ; therapy ; Infant, Newborn ; Infant, Newborn, Diseases ; prevention & control

In this study, we intend to confirm our hypothesis that cold inducible RNA-binding protein (CIRP) can inhibit neuronal apoptosis through suppressing the formation of oxygen free radicals under hypothermia. Primary rat hippocampal neurons were isolated and cultured in vitro, and were divided into five groups: (1) normal control group (37 °C), (2) cells infected by empty viral vector group, (3) CIRP over-expressed group, (4) CIRP knock-down group, and (5) hypothermia control group. Cells in groups 2-5 were cultured under 32 °C, 5% CO2. Apoptosis of hippocampal neurons were detected by Annexin V-FITC/PI staining and flow cytometry; Expression of CIRP was determined by Western blot; Redox-related parameters (T-AOC, GSH-Px, SOD, MDA) were detected by ELISA kits. Results showed that CIRP expression levels were significantly increased (P < 0.01) and the apoptotic rates were significantly decreased (P < 0.01) in hypothermia control group and CIRP over-expressed group when compared with normal control group. On the other hand, the apoptotic rate was significantly increased (P < 0.05) in CIRP knock-down group compared with that in hypothermia control group. The levels of redox parameters in hypothermia control group and CIRP over-expressed group were significantly changed in comparison with those in normal control group, CIRP knock-down group and empty viral vector infected group, respectively (P < 0.05 or P < 0.01). These results suggest that up-regulation of CIRP by hypothermia treatment can protect the neuron from apoptosis through suppressing the formation of oxygen free radicals.
Animals ; Apoptosis ; Cells, Cultured ; Cold Shock Proteins and Peptides ; metabolism ; Cold Temperature ; Hippocampus ; cytology ; Hypothermia ; Neurons ; cytology ; Oxidation-Reduction ; RNA-Binding Proteins ; metabolism ; Rats ; Up-Regulation

OBJECTIVETo investigate the protective effect of hypothermia combined with dexamethasone on spermatogenesis and the expression of intercellular adhesion molecule 1 (ICAM1) after testicular torsion-detorsion.

METHODSWe made unilateral testicular torsion models in 100 pubertal male Sprague-Dawley rats by 720 degree torsion of the left testis and then randomly divided them into four groups of equal number to be treated with normal temperature + physiological saline (group A), hypothermia + physiological saline (group B), normal temperature + dexamethasone (group C), and hypothermia + dexamethasone (group D). After 48 hours, we collected the testes, observed pathological changes of the testicular tissue by HE staining under the light microscope, detected the apoptosis of spermatogenic cells by TUNEL, and determined the expression of ICAM1 by Western blot.

RESULTSHE staining showed different degrees of testicular tissue injury in the four groups of rats, most obvious in group A, but mild in the other three. The ICAM1 protein expression was significantly higher in group A (0.68 +/-0. 03) than in B (0. 49 +/- 0. 06, P <0. 05) , C (0. 46 +/- 0. 09, P < 0.05) , and D (0.17 +/- 0.08, P <0.01). The nuclei were deep brown or brown. Lots of apoptotic spermatogenic cells were seen in the torsion testis of group A, with a significantly higher apoptosis index ( [33. 13 +/- 3.21 ]%) than in B ( [ 17. 12 +/-5.23 ]%, P < 0.05), C ([14.13 +/- 2.03]%, P <0.05), and D ([9.05 +/- 1.03]%, P <0.01).

CONCLUSIONHypothermia combined with dexamethasone can protect the testis from injury as well as the reproductive function of the testis after testicular torsion-detorsion and reduce the expression of ICAM1.

Animals ; Dexamethasone ; pharmacology ; Disease Models, Animal ; Hypothermia, Induced ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; physiopathology ; Spermatogenesis ; drug effects

OBJECTIVESTo observe the protective effect of retrograde venous perfusion of cryogenic liquid via accessory hemiazygos vein and treated with resveratrol on spinal cord injury and evaluate the expression changes of microtubule-associated protein 2 (MAP-2) after spinal cord ischemia reperfusion injury (SCII) in swine.

METHODSEighteen swine were divided into 3 groups: group I/R (n = 6, operation group), group CL (n = 6, retrograde venous perfusion of cryogenic liquid), group CL+Res (n = 6, retrograde venous perfusion of cryogenic liquid and treated with resveratrol after ischemia). In the group I/R, the aorta was clamped for 60 minutes and then removed. In the group CL and CL+Res, 9 g/L cold (4 °C) saline solution (perfusion rate, 16.65 ml/min) was infused into the accessory hemiazygos vein during ischemia.In the group CL+Res, the swine were treated with resveratrol (10 mg/kg) after spinal cord ischemia. Arterial pressure, blood gas analysis and the spinal canal and nasopharyngeal temperature changes were monitored during the surgery. Nervous function were assessed at 6 hours, 1, 2 days, 1, 2, 4 weeks and MAP-2 expression were detected at 4 weeks after reperfusion by using Western blot analysis in spinal cord tissue.

RESULTSAfter operation 18 swine were all survival. Behavioral scores of all groups decreased until 1 week after reperfusion and increased as time went by. The scores of group CL and CL+Res were higher than group I/R (F = 8.612, 17.276 and 11.985, P = 0.035,0.011 and 0.023) at 6 hours, 1, 2 days, group CL+Res were higher than group CL(P = 0.021) at 1 days after surgery. After descending aortic cross clamping, the spinal canal and nasopharyngeal temperature were obviously decreased in all groups and dropped to the lowest at 60 minutes after ischemia and 20 minutes after reperfusion in group I/R and the other groups respectively(F = 23.187-55.029, P < 0.01).In group CL(0.54 ± 0.26) and CL+Res (0.66 ± 0.31), the MAP-2 expression were higher than group I/R(0.37 ± 0.18) (F = 9.381, P = 0.037) , and the level in group CL+Res was higher than in group CL (P = 0.021) .

CONCLUSIONRetrograde venous perfusion of cryogenic liquid via accessory hemiazygos vein and treated with resveratrol can relieve the ischemia-induced spinal cord injury in swine.

Animals ; Hypothermia, Induced ; Male ; Microtubule-Associated Proteins ; metabolism ; Reperfusion Injury ; therapy ; Spinal Cord ; blood supply ; Spinal Cord Injuries ; therapy ; Stilbenes ; therapeutic use ; Swine

Brain injury secondary to hypoxia-ischemia (HI) is one of the major causes of neonatal death and severe, long-term neurologic deficits in children. Aside from hypothermia, no established therapies exist. Although the specific mechanisms of hypothermic neuroprotection remain unclear, in part hypothermia suppresses a broad range of injurious factors involved in the both early or primary and late or secondary phases of cell damage and death during the HI injury. In particular, latent (early recovery) phase-a brief period of normal cerebral energetics between resuscitation/reperfusion and the secondary phase of impaired energy metabolism and injury - represents the effective window of opportunity for initiation of therapeutic hypothermia, and ameliorate the later secondary energetic decline, neuronal death and the subsequent neurodevelopmental disability. Randomized controlled studies and systemic reviews have demonstrated that moderate hypothermia (33-35degrees C of core body temperature) using systemic or whole body cooling and selective head cooling, started within 6 hours after birth and protracted for 72 hours, significantly improves survival and reduces neurologic impairment in term and near-term infants with moderate and severe HI encephalopathy. Throughout the world, therapeutic hypothermia is increasingly recommended and we should have the protocols, equipment and training to treat the newborns with moderate and severe HI encephalopathy with therapeutic hypothermia.
Brain ; Brain Injuries ; Child ; Energy Metabolism ; Head ; Humans ; Hypothermia ; Hypoxia-Ischemia, Brain ; Infant ; Infant, Newborn ; Neurologic Manifestations ; Neurons ; Parturition

OBJECTIVETo explore the effect of mild to moderate hypothermia on the expressions of apoptosis-related genes in the brain tissue of rats after cardiopulmonary resuscitation (CPR).

METHODSCPR models were established by asphyxia in 15 male SD rats, which were randomized equally into normal temperature group, 34 degrees celsius hypothermia group and 32 degrees celsius hypothermia group. The brain tissues of the rats were obtained after treatment for 12 h to observe the pathological changes. The expression of caspase-3 in cerebral cortex neurons was determined with immunohistochemistry, and the expressions of Bcl-2 and Bax were detected by Western blotting.

RESULTSCompared with normal temperature group, the two hypothermia groups (especially 32 degrees celsius group) showed significantly decreased expression of caspase-3 in the cortical neurons (P<0.05). Bcl-2 protein expression was significantly increased in the hypothermia groups, especially in 32 degrees celsius hypothermia group (P<0.05). There was no significant difference in Bax protein expression among the 3 groups.

CONCLUSIONMild hypothermia can relieve brain injury by down-regulating caspase-3 expression and up-regulating Bcl-2 protein expression to inhibit apoptosis of the brain neurons. Hypothermia at 32 degrees celsius offers better protection of the brain tissue than hypothermia at 34 degrees celsius.

Animals ; Apoptosis ; Cardiopulmonary Resuscitation ; Caspase 3 ; metabolism ; Cerebral Cortex ; metabolism ; pathology ; Hypothermia, Induced ; Male ; Neurons ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the protective effects of hypothermia combined with dexamethasone on the testis of rats after testicular torsion reduction and on the expression of eNOS and apoptosis of spermatogenic cells.

METHODSWe made unilateral testicular torsion models in 80 adolescent male Sprague-Dawley rats by 720 degrees torsion of the left testis, and then randomly divided them into four groups of equal number to be treated with normal temperature + physiological saline (group A), hypothermia + physiological saline (group B), hypothermia + dexamethasone (group C), and normal temperature + dexamethasone (group D). After 48 hours, we collected the testes, observed pathological changes of the testicular tissue by HE staining under the light microscope, determined the expression of eNOS by immunohistochemistry, and detected the apoptosis of spermatogenic cells by TUNEL.

RESULTSHE staining showed different degrees of testicular tissue injury in all the four groups of rats, most obvious in group A, while protective effect was observed in the other three groups. Immunohistochemistry revealed significantly more positive cells and higher positive staining intensity in the torsion (left) testis in group A than in B (P < 0.05), C (P < 0.01) and D (P < 0.01). The nuclei were deep brown or brown. Lots of apoptotic spermatogenic cells were seen in the torsion testis of group A, with a significantly higher apoptosis index (31.12 +/- 4.68) than in B (16.58 +/- 6.22) (P < 0.05), C (8.60 +/- 1.15) (P < 0.01) and D (13.52 +/- 3.06) (P < 0.01).

CONCLUSIONIschemia-reperfusion injury after testicular torsion reduction can increase the apoptosis of spermatogenic cells and decrease testicular reproductivity. Hypothermia combined with dexamethasone can protect the testis from injury as well as the reproductive function of the testis after testicular torsion reduction.

Animals ; Apoptosis ; Dexamethasone ; pharmacology ; Germ Cells ; cytology ; metabolism ; Hypothermia, Induced ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; pathology ; Spermatogenesis ; Testis ; metabolism ; pathology

BACKGROUNDDuring cardiac arrest, the gastrointestinal tract is sensitive to ischemia. Protection of the gastrointestinal tract is a critical factor in determining prognosis following cardiopulmonary resuscitation (CPR). This study seeks to determine the extent of gastrointestinal tract injury and the potential protective effect of inducing hypothermia following a porcine cardiac arrest model and CPR.

METHODSVentricular fibrillation was induced by programmed electrical stimulation in 16 male domestic pigs (n = 8 per group). Four minutes after ventricular fibrillation, CPR was performed. Pigs that successfully restored spontaneous circulation then received intravenous infusions of saline at either 4°C or room temperature to produce hypothermic and control conditions respectively. Serum diamine oxidase and gastrointestinal adenosine triphosphate enzyme activity were determined and histopathology of the gastrointestinal tract was performed by light microscopy and electron microscopy.

RESULTSSignificant injury of the gastrointestinal tract after CPR was found. Na(+)-K(+) and Ca(2+) adenosine triphosphate enzyme activity in the gastric tissue were significantly high in animals receiving hypothermia treatment compared to controls. Hypothermia also significantly reduced serum diamine oxidase after CPR compared to the control group. Moreover, severe injury sustained by the gastrointestinal tissue was significantly ameliorated under hypothermic conditions compared to controls.

CONCLUSIONSGastrointestinal injury and abnormal energy metabolism are strikingly evident following CPR. Hypothermia, which is induced by an infusion of 4°C saline, can rapidly reduce internal body temperature, improve energy metabolism, and ameliorate injury to the gastrointestinal mucosa after CPR.

Animals ; Cardiopulmonary Resuscitation ; adverse effects ; Disease Models, Animal ; Energy Metabolism ; Gastrointestinal Tract ; injuries ; Heart Arrest ; therapy ; Hypothermia, Induced ; methods ; Male ; Swine