Objective To investigate the effects of flurbiprofen pretreatment on the permeability of bloodbrain barrier in a rat model of global cerbral ischemia-reperfusion (I/R) injury. Methods Forty-five male SD rats weighing 300-350 g were randomly divided into 3 groups (n = 15 each): sham operation group (group S); global cerebral I/R group (group I/R); flurbiprofen 10 mg/kg + global cerebral I/R group (group F). Global cerebral ischemia was induced by 20 min occlusion of bilateral common carotid arteries combined with hypotension (MAP maintained at 35-45 mm Hg). In group F, flurbiprofen 10 mg/kg was injected iv at 15 min before ischemia. Evans blue 3 ml/kg was injectcd iv at 24 h of reperfusion, then the rats were sacrificed and their brains were immediately removed for determination of the apoptosis rate, brain water content, Evans blue content, TNF-α, IL-1β and IL-10 content, and microscopic examination. Results The apoptosis rate, brain water content, Evans blue content, and TNF-α, IL-1β and IL-10 content were significantly higher in group I/R and F than in group S (P ＜ 0.05 or 0.01).The apoptosis rate, brain water content, and Evans blue content and TNF-α and IL-1β content were significantly lower, while IL-10 content was higher in group F than in group I/R (P ＜ 0.01). Global cerbral I/R-induced changes were significantly attenuated in group F. Conclusion Pretreatment with flurbiprofen can protect bloodbrain barrier against cerebral I/R injury by inhibition of the inflammatory reaction.

Objective:To evaluate the changes in the blood-cerebrospinal fluid barrier in postoperative delirium rats.Methods:One hundred and forty-seven healthy female Sprague-Dawley rats, aged 3 months, weighing 240-300 g, were divided into 3 groups ( n=49 each) using a random number table method: control group (group C), anesthesia group (group A) and postoperative delirium group (group P). Group C received no treatment.Group A received 2-h anesthesia with 1.4% isoflurane.Group S underwent an exploratory laparotomy under 1.4% isoflurane anesthesia.The behaviors of rats in each group were tested at 24 h before surgery and 6, 9 and 24 h after surgery using buried food test, open field test and Y maze test.Sodium fluorescence was injected through the tail vein at 6, 9 and 24 h after surgery.Then the rats were sacrificed, the choroid plexus (CP) was obtained, and cerebrospinal fluid (CSF) of bilateral cerebral ventricles was collected, and the expression of ZO-1, occludin, claudin1, E-cadherin and VE-cadherin in CP was detected using Western blot.FITC-dextran 10, 40 and 70 kDa was injected through the tail vein at 6 h after surgery, and then CSF was collected for determination of the concentrations of NaFI, 10, 40 and 70 kDa fluorescein isothiocyanate labeled dextran (FITC-dextran) in CSF by fluorescence spectrophotometry.CP was obtained to observe the morphology of choroid plexus epithelial cells (CPECs) of bilateral cerebral ventricles with a transmission electron microscope. Results:Compared with group C and group A, the latency to eat food in buried food test was significantly prolonged, the time of staying at the central region was shortened, the percentage of the number of entries into novel arm and percentage of time of staying at novel arm in Y maze test were decreased, the freezing time in open field test was shortened, the expression of ZO-1, occludin and claudin1 in CP was down-regulated, the concentrations of NaFI and 10 kDa and 40 kDa FITC-dextranin CSF were increased ( P<0.05 or 0.01), the CPECs arranged at random and loose, the microvilli of CPECs were absent, the tight junction was blurred, and the gap became wider in group P. Conclusion:The occurrence of postoperative delirium is related to the change in blood-cerebrospinal fluid barrier.

Objective To evaluate the effect of dexmedetomidine on NF-κB activity in ventilator-induced lung injury in dogs.Methods Thirty healthy 5-year-old Beagles,weighing 10-12 kg,were randomly assigned into 5 groups (n =6 each) using a random number table:control group (group C),mechanical ventilation (MV) group and 3 different concentrations of dexmedetomidine groups (groups DEX1-3).The animals were anesthetized with pentobarbital sodium,ketamine and atropine.The rats were tracheostomized and spontaneous breathing was maintained in group C,while the rats were tracheostomized and mechanically ventilated for 4 h in MV and DEX1-3 groups.Tracheal intubation was performed in all the groups.The concentration of O2 inhaled was set at 50％,respiratory rate at 15 bpm,tidal volume at 20 ml/kg,and positive end-expiratory pressure at 2 cm H2 O.A loading dose of dexmedetomidine 0.5,1.0 and 2.0 μg/kg was infused over 20 min before intubation,followed by continuous infusion (lasting for4 h) at a rate of 0.5,1.0 and 2.0μg·kg-1 ·h-1 in DEX1-3 groups,respectively.Blood samples were taken from the femoral artery at baseline state,and 1,2 and 4 h of MV for detection of PaO2.The animals were sacrificed after 4 h of MV.The lungs were removed for determination of W/D lung weight ratio (W/ D ratio),myeloperoxidase (MPO) activity,NF-κB p65 expression (by Western blot) and TNF-α mRNA expression (by RT-PCR) in lung tissues.Results Compared with group C,W/D ratio,MPO activity,and expression of NFκB p65 and TNF-α mRNA were significantly increased in group MV (P ＜ 0.05).Compared with group MV,MPO activity,and expression of NF-κB p65 and TNF-α mRNA were significantly decreased,and no significant change was found in W/D ratio in DEX2 and DEX3 groups (P ＜ 0.05).Conclusion Dexmedetomidine can inhibit NF-κB activity and reduce inflammatory responses,thus improving ventilator-induced lung injury in dogs.

Objective To evaluate the effects of dexmedetomidine or dezocine alone or the combination of the two agents on the emergence agitation in patients undergoing thoracic surgery.Methods Eighty ASA Ⅰ or Ⅱ patients,aged 18-64 yr,weighing 48-75 kg,scheduled for elective thoracic surgery under general anesthesia,were randomly divided into 4 groups (n =20 each):control group (group C),dexmedetomidine group (group DEX),dezocine group (group DEZ) and dexmedetomidine + dezocine group (group DEX + DEZ).In group DEX,dexmedetomidine 0.5 μg/kg was injected intravenously at 15 min before induction of anesthesia,followed by continuous infusion of dexmedetomidine at 0.4 μg· kg-1 · h-1 until beginning of chest closure.Dezocine 0.1 mg/kg was injected intravenously after beginning of chest closure in group DEZ.In group DEX + DEZ,dexmedetomidine 0.5 μg/kg was injected intravenously at 15 min before induction of anesthesia,followed by continuous infusion of dexmedetomidine at 0.4 μg·kg-1 ·h-1 until beginning of chest closure and then dezocine 0.1 mg/kg was injected intravenously.While the equal volume of normal saline was given instead in group C.Flurbiprofen axetil 50 mg was injected intravenously at beginning of skin closure in each group.Venous blood samples were collected for determination of plasma C-reactive protein (CRP),TNF-α and IL-10 levels at 10 min before induction of anesthesia (T1),before skin closure (T2),immediately after extubation (T3) and 15 min after extubation (T4).Side effects such as agitation during emergence from anesthesia were recorded.Sedation was assessed using Ramsay score.Results Compared with group C,the levels of plasma CRP and TNF-α at T2-4 and ratio of TNF-α/IL-10 were significantly decreased,the levels of IL-10 were increased at T2-4,the degree and incidence of agitation were decreased,and Ramsay score was increased in the other three groups (P ＜ 0.05).Compared with groups DEX and DEZ,the levels of plasma CRP and TNF-α at T2_4 and ratio of TNF-α/IL-10 were significantly decreased,the levels of IL-10 were increased at T2-4,and the degree and incidence of agitation were decreased in group DEX + DEZ (P ＜0.05).No side effects such as hypotension,bradycardia,respiratory depression,nausea and vomiting were observed in the four groups.Conclusion Dexmedetomidine or dezocine alone or combination of the two agents can decrease the degree and occurrence of emergence agitation and inhibit the inflammatory response simultaneously,and the combination of the two agents provides better efficacy than either alone in patients undergoing thoracic surgery.

Objective: To evaluate the effect of irisin preconditioning on global cerebral ischemia-reperfusion (I/R) injury in rats. Methods: Thirty-six healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were divided into 3 groups (n=12 each) using a random number table method: sham operation group (group S), global cerebral I/R group (group I/R) and irisin preconditioning group (group I). Global cerebral I/R was induced by occlusion of bilateral common carotid arteries combined with hypotension (MAP maintained at 35-45 mmHg) in anesthetized rats.At 30 min before ischemia, irisin 10 μg/kg (diluted to 10 μg/ml in normal saline) was intravenously injected in group I, and the equal volume of normal saline was intravenously injected in S and I/R groups.Morris water maze test was performed at day 3 of reperfusion to assess the cognitive function.Rats were sacrificed after the end of morris water maze test, and brains were removed for determination of histopathologic changes in hippocampal CA1 region (using HE staining), neuronal apoptosis in hippocampal CA1 region (Tunel staining), glial fibrillary acidic protein (GFAP) expression (by Western blot), myeloperoxidase (MPO) activity in the hippocampal tissues (by colorimetric assay), and contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in hippocampal tissues (by enzyme-linked immunosorbent assay). The cell necrosis rate and apoptotic rate were calculated. Results: Compared with group S, the escape latency was significantly prolonged on 1-5 days in group I/R and on 1-3 days in group I, the time of staying at 1st quadrant was significantly shortened, the cell necrosis rate and apoptotic rate were increased, the expression of GFAP was up-regulated, and the activity of MPO and contents of TNF-α and IL-1β were increased in I/R and I groups (P<0.05 or 0.01). Compared with group I/R, the escape latency was significantly shortened on 1-5 days, the time of staying at 1st quadrant was prolonged, the cell necrosis rate and apoptotic rate were decreased, the expression of GFAP was down-regulated, and the MPO activity and contents of TNF-α and IL-1β were decreased in group I (P<0.05 or 0.01). Conclusion Irisin preconditioning can reduce the global cerebral I/R injury in rats, and the mechanism may be related to inhibiting activation of astrocytes in hippocampus and reducing inflammatory responses.

Objective To explore the DNA expression of human cytomegalovirus (HCMV) in glioma and the association between HCMV infection and prognosis of glioma patients.Methods Used for this study were 89 specimens of glioma which had been surgically ablated and pathologically confirmed from the patients between January 2007 and December 2016 at Department of Neurosurgery,The First Affiliated Hospital of Zhengzhou University,and Department of Neurosurgery,The First Hospital of China Medical University.Of them,32 belonged to WHO grade Ⅱ,31 to WHO grade Ⅲ and 26 to WHO grade Ⅳ.Ten specimens of normal brain tissue were excised as controls from the contemporary patients receiving resection for essential epilepsy.Nested PCR was used to analyze the DNA expression of HCMV in the glioma tissue and normal brain tissue,and in the peripheral blood from the glioma and control patients.Prognosis of the glioma patients was evaluated using the Kaplan-Meier survival analysis.Results The DNA expression of HCMV was positive in 46 of the 89 specimens of glioma,involving 14 cases of WHO grade Ⅱ,16 ones of WHO grade Ⅲ and 16 ones of WHO grade Ⅳ.The DNA expression of HCMV was negative in all the 10 specimens of normal brain tissue.There was a significant difference in the DNA expression of HCMV between the glioma tissue and normal brain tissue (P=0.002).The HCMV DNA was measured in the peripheral blood from 26 glioma patients,involving 10 cases of WHO grade Ⅱ,8 ones of WHO grade Ⅲ and 8 ones of WHO grade Ⅳ.No HCMV DNA was detected in the peripheral blood from the 10 control patients.There was a significant difference between the brain glioma and control groups in gene expression of HCMV in peripheral blood (P=0.048).There were no significant differences in the survival rate between the patients with positive or negative DNA expression of HCMV in the glioma tissue or in the peripheral blood from the glioma and control patients (x2=1.849,P=0.174;x2=0.082,2=0.774).Conclusion HCMV infection may play an active role in pathogenesis and development of glioma.

Objective To evaluate the effect of quercetin pretreatment on the permeability of blood-brain barrier in a rat model of global cerebral ischemia-reperfusion ( I∕R). Methods Sixty-three clean-grade healthy male Sprague-Dawley rats, weighing 300-350 g, aged 4-5 months, were divided into 3 groups (n＝21 each) using a random number table method: sham operation group ( group S), group I∕R and quercetin pretreatment group ( group Q). Global cerebral I∕R was induced by occlusion of bilateral common carotid arteries combined with hypotension ( mean arterial pressure was maintained at 35-45 mmHg) in chloral hydrate-anesthetized rats. Quercetin 25 μmol∕kg was injected intraperitoneally twice a day for 3 consecutive days starting from 3 days before establishment of the model in group Q, while the e-qual volume of normal saline was given instead at the corresponding time points in group S and group I∕R, respectively. The animals were sacrificed at 24 h of reperfusion and brains were removed to determine the brain water content, Evans blue ( EB) content and expression of occludin protein in cerebral cortex ( by Western blot) and to observe the ultrastructure of blood-brain barrier. Results Compared with group S, the brain water content and EB content were significantly increased, the expression of occludin protein was down-regulated (P＜0. 05), and the injury to ultrastructure of blood-brain barrier was accentuated in I∕R and Q groups. Compared with group I∕R, the brain water content and EB content were significantly de-creased, the expression of occludin protein was up-regulated (P＜0. 05), and the injury to ultrastructure of blood-brain barrier was significantly attenuated in group Q. Conclusion Quercetin pretreatment can de-crease the permeability of blood-brain barrier and attenuate brain edema, and the mechanism may be related to up-regulated expression of occludin protein in a rat model of global cerebral I∕R.

Objective:To investigate the effect of oxycodone hydrochloride injection pretreatment on focal cerebral ischemia-reperfusion injury in rats.Methods:Seventy-two male SD rats weighing 200-250 g were randomly divided into 3 groups( n=24 each group): sham operation group (sham group), focal cerebral I/R group (I/R group), and oxycodone hydrochloride injection group (Oxy group). Focal cerebral I/R was induced by middle cerebral artery occlusion for 2 h followed by reperfusion. In the Oxy group, oxycodone hydrochloride 0.5 mg/kg was injected iv at 5 min before ischemia. While the same volume of saline (1 mL) was injected in the sham group and I/R group. The neurological deficit score (NDS) was assessed at 24 h of reperfusion, the rats were then sacrificed, and their brains were immediately removed for determination of brain water content and the infarct volume, and the histopathological changes were observed after HE staining. The levels of cytokines (TNF-α, IL-1β and IL-10) in the ischemia cortex were quantified by ELISA. MPO activity in the ischemia cortex was assessed. Western blot was used to detect the expression of NF-κB in the ischemia cortex. The data were analyzed using SPSS 20.0 software, multiple-group comparisons were performed using one-way ANOVA, and LSD- t test was used for pairwise comparison between groups. A P<0.05 was considered statistically significant different. Results:Compared with the sham group, NDS, brain water content, relative infarction volume and rate of nerve cell necrosis were significantly increased in the I/R and Oxy groups (all P<0.05). Levels of TNF-α, IL-1β, IL-10, NF-κB and the activities of MPO were increased in the ischemia cortex (all P<0.05). Compared the Oxy group with the I/R group, NDS, brain water content, relative infarction volume and rate of nerve cell necrosis were significantly decreased [(1.7±0.9) vs (2.6±1.1);(79.2±2.4)% vs (84.7±4.2)%; (23.0±5.4)% vs (34.8±6.0)%; (25.2±12.4)% vs (54.8±14.8)%, all P<0.05]. The levels of TNF-α, IL-1β, relative expression of NF-κB, and the activities of MPO were significantly decreased in the ischemia cortex [(4.4±1.2) pg/mg vs (6.5±1.2) pg/mg; (5.4±0.7) pg/mg vs (7.8±0.8) pg/mg; (0.83±0.11) vs (1.23±0.33); (0.27±0.09) U/g vs (0.36±0.14) U/g, all P<0.05] , while the concentration of IL-10 was significantly increased [(20.9±4.5) pg/mg vs (9.2±1.6) pg/mg, t=6.036, P=0.000 1]. Conclusions:Oxycodone hydrochloride can attenuate focal cerebral I/R injury through inhibiting NF-κB activity.

[Abstract] Objective: To evaluate the expression level of FOXD1 in glioma tissues of different grades, and to investigate the correlation between the expression of FOXD1 and the prognosis of glioma patients. Methods: The tumor tissues were collected from 40 glioma patients, who received surgical treatment in the neurosurgery department of the First Hospital of China Medical University from September 2014 to February 2015; Seven non-tumor tissues obtained from patients underwent internal decompression for traumatic brain injury were used as controls. The FOXD1 expression in glioma and non-tumor brain tissues was analyzed by qRT-PCR and IHC, and the correlations between clinical pathological features of glioma patients and FOXD1 expression level were analyzed. Furthermore, the Kaplan-Meier method was used to analyze the relationship between FOXD1 expression and survival time of patients. In addition, the expression of FOXD1 in glioma tissues and its relationship with patients’prognosis were confirmed by the data from GEO (GSE4290, GSE2223) and Rembrandt database. Results: qRT-PCR showed that the FOXD1 mRNA expression in glioma tissues of WHO grade IV was significantly higher than that of non-tumor brain tissues and glioma tissues of WHO grade II (P<0.01). German immunohistochemical score (GIS) was used to evaluate the immunohistochemical staining intensity, and the relationship between FOXD1 expression and clinical pathological features was analyzed. The results showed that FOXD1 in glioma tissues was related to WHO phathological grade level (χ2=11.73， P<0.01). There was statistically significant difference between the survival time of FOXD1 high expression group and FOXD1 low expression group (P=0.043). The data from GEO data base (GSE4290, GSE2223) and Rembrandt datasets showed that glioma tissues have a higher FOXD1 mRNA expression level than normal brain tissues, and the elevated expression of FOXD1 mRNA was negatively associated with the survival time of glioma patients. Conclusion: FOXD1 was highly expressed in glioma tissues, and the expression level of FOXD1 was increased as the pathological grade increases. The elevated expression of FOXD1 was related with the poor survival of glioma patients.

Preoperative sleep loss can amplify post-operative mechanical hyperalgesia. However, the underlying mechanisms are still largely unknown. In the current study, rats were randomly allocated to a control group and an acute sleep deprivation (ASD) group which experienced 6 h ASD before surgery. Then the variations in cerebral function and activity were investigated with multi-modal techniques, such as nuclear magnetic resonance, functional magnetic resonance imaging, c-Fos immunofluorescence, and electrophysiology. The results indicated that ASD induced hyperalgesia, and the metabolic kinetics were remarkably decreased in the striatum and midbrain. The functional connectivity (FC) between the nucleus accumbens (NAc, a subregion of the ventral striatum) and the ventrolateral periaqueductal gray (vLPAG) was significantly reduced, and the c-Fos expression in the NAc and the vLPAG was suppressed. Furthermore, the electrophysiological recordings demonstrated that both the neuronal activity in the NAc and the vLPAG, and the coherence of the NAc-vLPAG were suppressed in both resting and task states. This study showed that neuronal activity in the NAc and the vLPAG were weakened and the FC between the NAc and the vLPAG was also suppressed in rats with ASD-induced hyperalgesia. This study highlights the importance of preoperative sleep management for surgical patients.
Rats ; Animals ; Hyperalgesia/metabolism* ; Sleep Deprivation/metabolism* ; Rats, Sprague-Dawley ; Periaqueductal Gray/pathology* ; Proto-Oncogene Proteins c-fos/metabolism* ; Pain, Postoperative/pathology*