BACKGROUND: Hyaluronan-methylcellulose hydrogel cannot only be conjugated with short peptide sequences and growth factors to achieve sustained release, but also has a role in blocking dural defects and reducing inflammation. It is an ideal biomaterial for the treatment of spinal cord injury. OBJECTIVE: To investigate the effect of neurotrophin-3 modified hyaluronan-methylcellulose (HAMC-NT-3) hydrogel on the recovery of neurological function in rats with spinal cord injury. METHODS: Fifty-four female Sprague-Dawley rats (provided by the Experimental Animal Center of the Academy of Military Medical Sciences in China) were randomly divided into three groups (n=18 per group) . The sham group only underwent T10 laminectomy. In the model group and the experimental group, an aneurysm clip was used to establish spinal cord injury models after T10 laminectomy. The experimental group was locally injected with HAMC-NT-3 hydrogel. The Basso Beattie Bresnahan function scoring was performed at 1 day, 1, 2, 3, 4, 5, 6, 7, and 8 weeks after surgery. The inclined plane test was performed at 4, 6 and 8 weeks after surgery to evaluate the recovery of hindlimb motor function. ELISA was used to detect the concentrations of inflammatory factors in the spinal cord at 1 week after surgery. Immunohistochemical staining was used to observe the area of syringomyelia, glial fibrillary acidic protein expression and nerve regeneration at 8 weeks after surgery. RESULTS AND CONCLUSION: (1) The Basso Beattie Bresnahan scores of the model group and the experimental group were lower than those of the sham group at various time points after surgery (P < 0.05) . The Basso Beattie Bresnahan scores of the experimental group were higher than those of the model group at 4-8 weeks after surgery (P < 0.05) . (2) In the inclined plane test, the maximum inclined angles of the model group and the experimental group at each time point after surgery were lower than that of the sham group (P < 0.05) . The maximum inclined angles of the experimental group at 6 and 8 weeks after surgery were higher than those of the sham group (P < 0.05) . (3) The concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-6 and interleukin-10 in the experimental group and the model group were higher than those in the sham group (P < 0.05) . The concentrations of tumor necrosis factor-α, interleukin-1β and interleukin-6 in the experimental group were lower than those in the model group (P < 0.05) . The concentration of interleukin-10 in the experimental group was higher than that in the model group (P < 0.05) . (4) Immunohistochemical staining showed that the expression levels of glial fibrillary acidic protein in the experimental group and the model group were higher than those in the sham group, while the expression of glial fibrillary acidic protein in the experimental group was lower than that in the model group. The area of syringomyelia in the experimental group was smaller than that in the model group (P < 0.05) . These results indicate that local injection of HAMC-NT-3 hydrogel can effectively inhibit inflammation as well as astrocyte activation and proliferation, reduce fibrous scar formation, and promote the protection of nerve tissue and the recovery of hindlimb motor function after spinal cord injury.

Objective To explore the effect of Dexmedetomidine (Dex) on acute brain edema in mice in condition with targeted temperature management (TTM) following traumatic brain injury (TBI).Methods A total of 180 male C57BL/6J mice were divided into control group,sham operation group,TBI group,TBI + Dex group,TBI + TTM group,and TBI + Dex + TTM group according to the random number table (n =30 per group).The sham operation group only opened the bone window but did not hit it,and the control group did not open the bone window.The TBI + Dex,TBI + TIM,and TBI + Dex + TTM groups were intraperitoneally injected with Dex (60 μg/kg once every 2 h for 3 times) and/or hypothermia after TBI.The brain tissue injury volume,EB extravasation and brain water content of each group were determined by toluidine blue,Evans blue staining and dry-wet weight method at 24 hours after injury.Real-time quantitative PCR and Western blot were used to detect the expression of Claudin-5 in the injured brain tissue.At 24,48,and 72 hours after injury,the neurological deficiency degree was assessed using the modified neurological severity scores (mNSS).Results Compared with the sham operation group,TBI mice showed significant increase in brain tissue injury volume [(0.49 ± 0.04)mm3 vs.(1 1.57 ± 1.01)mm3],blood-brain barrier permeability [(16.4 ± 0.8) μg/g vs.(54.3 ± 1.7) μg/g],brain tissue water content [(76.7 ± 0.9) ％ vs.(83.1 ± 0.8) ％],and mNSS score [(1.6 ± 0.7) points vs.(13.4 ± 0.7) points] at 24 hour after TBI (all P ＜ 0.01).However,Dex or TTM treatment reduced brain tissue injury volume [(7.20±0.18)mm3 and (5.94 ±0.18)mm3],blood-brain barrier permeability [(32.7 ± 1.2) μg/g and (27.6 ± 1.0) μg,/g],brain tissue water content [(78.5 ± 0.4) ％ and (78.2 ± 0.6) ％],and neurological function [mNSS:(7.3 ± 1.1) points and (5.8 ± 1.3) points] (all P＜0.01).Moreover,Dex + TTM group showed better neuroprotection [reduced brain tissue injury volume:(3.92 ± 0.05) mm3,reduced BBB permeability:(21.6 ± 0.7) μg/g,reduced brain water content:(77.7 ±0.3)％,and reduced mNSS:(4.3 ± 1.2) points] compared with Dex or TTM alone (all P ＜ 0.01).Additionally,the mRNA expression of Claudin-5 (0.23 ± 0.01) decreased significantly at 24 hours after TBI compared with sham group (0.93 ± 0.04,P ＜ 0.01),but Dex or TTM could increase the expression of Claudin-5 (0.47 ± 0.01,and 0.54 ± 0.09) compared with TBI group (P ＜0.01),especially that of TBI + Dex + TTM group (0.64 ± 0.02,P ＜ 0.01).Furthermore,the protein expression of Claudin-5 was in accordance with the result of its mRNA expression.Conclusion Dex in condition with targeted temperature management can up-regulate Claudin-5 expression in early TBI,protect the integrity of blood-brain barrier,attenuate acute brain edema and neurological damage,and improve neurological function recovery.

Objective: To investigate the neuroprotective effect of Ghrelin on traumatic brain injury (TBI) in mice. Methods: TBI model of C57BL / 6 mice was established by electronic cortical impact instrument (eCCI). According to the random figure table method, twenty-four mice were randomly divided into sham group(Sham group), TBI group and Ghrelin intervention group(Ghrelin group) with 8 mice in each group. The model of TBI was established in TBI group and Ghrelin group.The mice in Ghrelin group was injected intraperitoneally 0.5 g/kg before and 1 h after injury respectively. And the mice Sham group and TBI group were injected with the same amount of normal saline. The changes of cerebral blood perfusion (CBP) were monitored in real time by laser speckle contrast analysis(LSCI), the changes of neuroelectrophysiology were observed by monitoring motor evoked potential (MEP), and the status of neurological deficit was evaluated by modified neurological deficit score (mNSS). Results: Compared with Sham group, the mice in TBI group had significantly lower cerebral blood perfusion(CBP) (t=-12.36, P<0.01), longer latency and lower amplitude of motor evoked potential (MEP) (t=5.03, -11.55, all P<0.01), and significantly higher mNSS scores (t=9.34, P<0.01). However, compared with the TBI group, the cerebral blood perfusion(CBP) of Ghrelin group increased significantly at 12 h after TBI((196.87±17.36) PU/mm² vs (123.62±8.04)PU/mm², t=10.45, P<0.01), while the latency of MEP decreased((5.30±0.33)ms vs (6.80±0.97)ms, t=-5.01, P<0.01), the amplitude of MEP increased((2.21±0.16)mV vs (1.27±0.27)mV, t=9.65, P<0.01). And compared with the TBI group, the neurological deficit score of Ghrelin group decreased significantly at 24 h after TBI((4.9±1.2) vs (8.4±2.6), t=-3.87, P<0.01). Conclusion Ghrelin exhibits a significant neuroprotective role by increasing cerebral blood flow perfusion, reducing the degree of neurological deficit and promoting motor function recovery in TBI mice.

Objective To investigate the effect of Ghrelin on gastrointestinal motility after traumatic brain injury (TBI).Methods A total of 72 adult male SD rats were randomly divided into sham operation group (n =8),TBI group (n =32) and Ghrelin group (n =32),according to the random number table.In the sham operation group,the scalp was sutured after craniotomy and sterilization,without any strike.In the TBI group,after intraperitoneal anesthesia,the skull was opened and the electric cortical contusion impactor was used to strike the center of bone window at the depth of 3 mm and the rate of 5 m/s.The duration of hitting the lowest point was 200 ms.In the Ghrelin group,20 μg/kg of Ghrelin was injected into the rat via the tail vein 30 minutes after injury.The modified neurologicalseverity score (mNSS),percentage of water content in feces and percentage of gastric contents in body weight at 6,24,48 and 72 hours after operation in each group were measured.The stomach,the small intestine 15 cm from ileocecal junction,ileocecal junction (about 3 cm in the proximal ileal loops,about 3 cm in the distal ileal loops,and 3 cm colon loops) were taken out to prepare the electron microscopy section and observe the microscopic changes of the gastrointestinal mucosa.Results The mNSS in the TBI and Ghrelin groups was higher than that in the sham operation group after 24,48 and 72 hours (P ＜0.01).The mNSS in the TBI group was higher than that in the Ghrelin group after 24,48 and 72 hours (P ＜0.01).In the sham operation group,the intestinal wall was pink.In the TBI group,gastric dilatation and thinner wall with pale or dark red color were seen,and small intestine cavity expansion with dark color and even congestion were observed.There was much mucus in the intestinal wall.The Ghrelin group improved obviously than the TBI group after 6 hours.Compared with the Ghrelin group,the percentage of fecal water content in the TBI group decreased significantly after 24 hours (P ＜ 0.05),and the decrease rate dropped with time.Obvious delayed gastric emptying occurred (P ＜ 0.05),and the percentage of gastric contents in body weight demonstrated downtrend.The changes of gastric mucosa were as follows:the chief cells in the gastric glands were observed 72 hours after TBI in the TBI group,and scattered short microvilli were seen in the cell surface.The cytoplasm protruded into the glandular cavity,and a large number of rough endoplasmic reticulum could be seen in the cytoplasm,with irregular arrangement.Medullary bodies could be seen inside the mitochondria which swelled locally.Abundant endocrine granules were seen in the cytoplasm.Mitochondria were scattered and swollen,and mitochondria cristae became shorter and fewer,which contained medullary bodies.The Ghrelin group improved obviously than TBI group after 72 hours in terms of gastric mucosa changes.With respect to cecum mucosa,in the TBI group 72 hours after TBI,severe edema of the cecum absorption epithelium,obvious dilation of the rough endoplasmic reticulum,expansion of the free water gap inside the cell,and local decrease of the microvilli at the top of the cell were observed.Abundant microvilli were seen in the cecum absorption epithelium and cell top.The connection complex composed of tight connections,intermediate connections,and bridging particle connections could be seen between cells.The Ghrelin group improved obviously than TBI group after 72 hours in terms of cecum mucosa changes.Conclusions Ghrelin can improve gastrointestinal motility and protect gastrointestinal mucosa in rats after TBI.

Objective: To investigate the effects of Casein kinase 2-interacting protein 1 (CKIP-1) gene silencing on the proliferation of glioma cells U87-MG. Methods: The recombinant lentiviral vectors targeting CKIP-1 gene or negative control were constructed and then used to infect glioma U87-MG cell line.The effects of knock-down on the mRNA or protein expression of CKIP-1 were evaluated by real-time qPCR and western blotting.Cell cycle was detected by the flow cytometry assay, and cell proliferation changes were evaluated by cell counting, MTT, and BrdU assay, respectively.Lastly, the colony formation was used to investigate the effect of CKIP-1 knock-down on the clone formation. Results: Compared with the group of Ctrl, CKIP-1 siRNA was observed to significantly inhibit CKIP-1 expression at the mRNA levels (Ctrl (1.01±0.13) vs CKIP-1 siRNA (0.23±0.02), P<0.01) and protein levels in the U87-MG cells.Also, CKIP-1 suppression mediated by RNAi decreased the ratio of G0/G1 phase (Ctrl (69.64±0.79) vs CKIP-1 siRNA(62.64±0.66), P<0.01), increased that of G2/M phase (Ctrl (8.36±0.52) vs CKIP-1 siRNA(13.87±2.90), P<0.05), and significantly inhibited the cell proliferation and clone formation (Colony number: Ctrl (25±2) vs CKIP-1 siRNA(2±1), P<0.05) of transfected U87-MG cells. Conclusion Knocking down the expression of CKIP-1 significantly inhibit cell proliferation in human U87-MG glioma cells, indicating that CKIP-1 is involved in the development of gliomas and could promote the cell proliferation.

Objective To explore the effects of humanistic care on preoperative negative emotions in patients undergoing eye surgery, so as to alleviate the negative emotion and promote the rehabilitation process of the operation. Methods From October 2016 to February 2017, totals of 120 patients who were undergoing eye surgery from each of the 2 wards of the ophthalmology department in the First Affiliated Hospital of Zhengzhou University were recruited in the research. All the patients were randomly divided into control group and observation group by coin tossing method. Routine nursing was applied in the control group, while on the basis of the control group, humanistic care was given to the observation group. The intervention effects were evaluated by the general information questionnaire, the Self-Perceived Burden Scale (SPBS), Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS). Results There was no statistical difference in negative emotions between the two groups before intervention (P>0.05). The scores of negative emotions all had a certain decrease after intervention. The scores of SPBS, SAS and SDS were (28.49±5.92), (45.49±6.49) and (41.22±5.62) in the observation group after intervention respectively; while those scores were (34.95±6.56), (58.57±8.56) and (54.53±6.45) in the control group respectively. The scores of the observation group were all lower than those of the control group, and the differences were statistically significant (t=10.49, 6.45, 7.34; P< 0.01). Conclusions Humanistic care can obviously reduce patients' self-perceived burden, anxiety, depression and improve patients negative emotions.

Objective To explore the effect and mechanism of necroptosis related proteins in middle cerebral artery occlusion (MCAO) induced brain ischemia/reperfusion injury in mice.Methods C57BL/6 mice were used to establish the brain ischemia/reperfusion injury model induced by MCAO.MCAO mice were treated with z-VAD.fmk (zVAD,1.1 g/kg),GSK'872 (0.7 g/kg) and combined intervention of zVAD and GSK'872,and neurological defect was evaluated by mNSS while brain infarct volume was measured by TTC staining.Western blot and immunofluorescence assay were used to detect protein expression and location of RIP1,RIP3 and MLKL,respectively.Results Neurological defect and brain infarction were caused by MCAO.Compared with MCAO group,zVAD,GSK'872 and the combined intervention alleviated neurological defect and reduced brain infarct volume significantly (P＜0.05 or P＜0.01).The protein levels of RIP3 and RIP1 MLKL were increased in mice of MCAO group,while GSK'872 and the combined intervention obviously downregulated the aforementioned protein expression [RIP1 (GSK'872:0.64± 0.02 vs MCAO:1.28±0.02,P＜0.01);RIP3 (GSK'872:1.08±0.02 vs MCAO:1.45±0.02,P＜0.01);MLKL (GSK'872:0.54±0.01 vs MCAO:1.00±0.01,P＜0.01)].However,zVAD only slightly reduced protein expression of MLKL (P＜0.05) but didn't change the protein expression of RIP1 and RIP3 (P＞0.05).Conclusion RIP1,RIP3 and MLKL are involved in the execution of necroptosis and contribute to the pathological progress of brain ischemia/reperfusion injury.

Objective To screen altered proteins of hippocampus in the stress-induced depression (STRID) rat model, and explore the potential molecular mechanism. Methods Twenty Sprague-Dawley rats were randomly divided into the control group and STRID group, 10 rats in each group. Chronic unpredictable mild stress (CUMS) methods including fasting for solids and liquids, electric foot-shock, reversing day and night, cold water swimming, cage tilt, scare stimulation and tail pinch were conducted on STRID rats with no repeats for 28 days to make up the depression animal model. The control group was normally fed during this period. After the stress stimulation, the hippocampus protein samples were used for two dimensional electrophoresis to screen the differentially expressed protein, and then mass spectrum identification and function analyze were conducted. Results Compared with the control group, 34 proteins were altered in STRID group. Among which, 18 were up-regulated, and 16 were down-regulated. The differentially expressed proteins mainly located in cytoplasm, mitochondrion, extracellular exosome and myelin sheath. The involved signaling pathways included metabolic pathway, oxidative phosphorylation pathway, and Alzheimer's disease, Parkinson's disease and Huntington's disease pathways. Conclusion The altered proteins and dysfunction of nerve signaling, and the excess of oxidative phosphorylation in hippocampus of STRID rats may be one of the pathogenesises.

Objective To explore the effects of different doses of dexmedetomidine on acute brain edema in mice with traumatic brain injury (TBI).Methods A total of 132 male C57BL/6J mice were randomly divided into six groups:control group (group C),sham-operation group (group Sham),traumatic brain injury group (group TBI),Dex 20 μg/kg (group D20),40 μg/kg (group D40),and 60 μg/kg (group D60),n=22 in each group.The TBI animal model was established by electric controlled cortical impactor (eCCI),then intraperitoneal injected by the administration of different doses of dexmedetomidine at 0,2 and 4 h after TBI.Twenty-four hours post-TBI,brain water content was measured by the dry-wet method,histological observation was performed using HE staining,and aquaporin 4 (AQP4) and NF-κB expression were detected using Western blot assay,respectively.Then,the modified neurological scale scores (mNSS) on 1,2,3,and 7 d and Morris water maze (MWM) test on 4,5,6 and 7 d post-TBI were used to evaluate the neurologic deficit of TBI mice.Results After traumatic brain injury,the mNSS scores,the escape latency,the brain water content and the expression of AQP4 and NF-κB increased significantly in group TBI (P＜0.01).Different doses of dexmedetomidine significantly reduced the mNSS scores,the escape latency,the brain water content and the expression of AQP4 and NF-κB (P ＜ 0.05 or P ＜ 0.01).And meanwhile dexrnedetomidine can lessen neuronal degeneration,and inflammation response.Additionally,the effect was remarkably in group D60 compared with group D20 (P ＜ 0.05 or P ＜ 0.01).Conclusion Dexmedetomidine can lessen brain edema and cognition impairment induced with traumatic brain injury,which is a dose-effect relationship within 20-60 μg/kg,and this effect may be related to the downregulation of AQP4 and NF-κB expression.

The refractory epilepsy refers to the epilepsy whose seizures couldn't be cured after using two kinds of correctly selected antiepileptic drugs which can be tolerated and enough dosage and duration of monotherapy or combination therapy.The pathogenesis of intractable epilepsy is complex,and there is no established theory at home and abroad.Recently,more and more attention has been paid to the correlation between mitochondrial dysfunction caused by oxidative stress and intractable epilepsy.Based on the summary of common treatment of refractory epilepsy and by searching the related literature at home and abroad,further investigation would be made to explore the diagnosis and treatment of intractable epilepsy theory in order to provide new strategies for diagnosis and treatment of intractable epilepsy.