Objective To evaluate the relationship between the levels of microRNA-125b (miR-125b) and miR-181c in cerebrospinal flnid (CSF) before joint replacement and postoperative delirium (POD) in elderly patients.Methods Fifty-two patients of hoth sexes,aged ≥ 65 yr,weighing 45-70 kg,of American Society of Anesthesiologists physical status Ⅰ-Ⅲ,scheduled for elcctive hip or knee replacement under spinal anesthesia,were included in the study.CSF was collected after successful puncture to measure the levels of miR-181c and miR-125b by fluorescent quantitative real-time polymerase chain reaction.The patients were divided into POD group and non-POD group using the Chinese reversion of Confusion Assessment Method on postoperative days 1 and 2.Results The incidence of POD was about 28％ in the patients underwent hip or knee replacement.Compared with non-POD group,the preoperative level of miR-181c in CSF was significantly increased (P＜0.05),and no significant change was found in the preoperative level of miR-125b in CSF in POD group (P＞0.05).Conclusion The level of miR-181c in CSF before joint replacement is related to POD in elderly patients,and the preoperative level of miR-181c in CSF is a risk factor for POD.

Objective To investigate the changes of miR-146a expression in serum,hippocam-pus and prefrontal cortex in postoperative cognitive dysfunction (POCD) mice.Methods Fifty healthy male C57BL/6 mice,aged 12-14 weeks,weighing 25-30 g,were randomly divided into two groups (n =25 each)using a random number table:control group (group C)and anesthesia plus sur-gery group (group AS).Mice in group AS underwent open tibial fracture of the left hind paw with in-tramedullary fixation in aseptic conditions under general anesthesia with 2.1% isoflurne.Ten mice in each group received the fear conditioning test (FCT)on the 1,3 and 7 days after anesthesia/surgery. The rest of mice were sacrificed 24 h before (baseline),and 6,12,24,48 h after anesthesia/surgery, and then the serum,prefrontal cortex and hippocampus were collected or removed for detection of the expression of miR-146a using quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results Compared with group C,the percentage of freezing time in contextual FCT was sig-nificantly decreased (P <0.05)in group AS,while no significant change in freezing time percentage was found in tone-cued FCT.In serum,compared with group C,miR-146a expression at 6,12,24, 48 h after anesthesia/surgery was significantly up-regulated in group AS (P < 0.05 );and in group AS,the expression of miR-146a was significantly decreased 6,24,48 h as compared to that at 12 h after anesthesia/surgery (P <0.05).In hippocampus,compared with group C,miR-146a expression at 6,12,24,48 h after surgery was significantly up-regulated in group AS (P <0.05);and in group AS,the expression of miR-146a at 6,48 h after surgery was significantly decreased as compared to that at 12 h after anesthesia/surgery (P <0.05).In prefrontal cortex,compared with group C,miR-146a expression at 24,48 h after surgery was significantly up-regulated in group AS (P <0.05);and in group AS,the expression of miR-146a was significantly increased at 48 h as compared to that at 24 h after anesthesia/surgery (P <0.05).Conclusion The expression of miR-146a in serum,hippocam-pus and prefrontal cortex in POCD mice was up-regulated,and changes of miR-146a expression may be related to the development of POCD.

Objective To evaluate the effect of exogenous miR-181b on postoperative cognitive dysfunction ( POCD) in mice. Methods A total of 108 clean-grade healthy male C57BL∕6 mice, weighing 25-30 g, were divided into 4 groups ( n=27 each) using a random number table method: control group ( group C) , POCD group, miR-181b agonist agomir group ( group Ag) , and agomir negative control group ( group AC) . POCD was induced by open tibial fracture in 2. 1％ isoflurane-anesthetized mice. A total vol-ume of miR-181b agomir 4μl ( 0. 25 nmol∕μl) was injected into bilateral hippocampi at 2 days before estab-lishing the model in group Ag. Agomir negative control solution 4μl was given at 2 days before establishing the model in group AC. Eighteen mice were selected at days 1, 3 and 7 after surgery, and contextual fear conditioning test was performed to assess cognitive function. Nine mice in each group were sacrificed at 6, 12 and 24 h after surgery ( 3 mice at each time point) , and the hippocampus was isolated for determination of miR-181b expression ( by quantitative real-time polymerase chain reaction) and interleukin-1β ( IL-1β) and tumor necrosis factor-α ( TNF-α) contents ( by enzyme-linked immunosorbent assay) . Results Com-pared with group C, the percentage of time spent freezing in the contextual fear conditioning test was signifi-cantly decreased at 1, 3 and 7 days after surgery, and the expression of miR-181b was down-regulated and the contents of IL-1β and TNF-α were increased at 6, 12 and 24 h after surgery in POCD and AC groups( P<0. 05) . Compared with POCD and AC groups, the percentage of time spent freezing in the contextual fear conditioning test was significantly increased 1, 3 and 7 days after surgery, and the expression of miR-181b was up-regulated and the contents of IL-1βand TNF-αwere decreased at 6, 12 and 24 h after surgery in group Ag ( P<0. 05). Conclusion Exogenous miR-181b can mitigate POCD, and the mechanism may be related to inhibiting inflammatory responses in the hippocampus of mice.

Objective To investigate the role of hippocampal cannabinoid receptor 2 (CB2R) in postoperative cognitive dysfunction (POCD) in mice.Methods Fifty-four healthy male C57BL/6 mice,aged 12-16 weeks,weighing 20-30 g,were divided into 3 groups (n=18 each) using a random number table:control group (group C),group POCD and POCD plus CB2R agonist JWH133 group (group POCD+J).The mice were subjected to an intramedullary fixation for tibial fracture under 1.5％ isoflurane anesthesia to establish POCD model.In group POCD+J,2 mg/kg JWH133 was injected intraperitoneally at a total volume of 10 ml/kg after emergence from anesthesia and then the injection was repeated once a day until the 7th day after surgery.Open field test was carried out on 1 day before surgery and 1,3 and 7 days after surgery to evaluate the locomotor activity.Fear conditioning test was carried out at 15 min after completion of open field test.Mice were sacrificed at 2 h after the end of behavioral test,and the hippocampus was harvested for determination of the expression of tumor necrosis factor-alpha (TNF-α),interleukin-1beta (IL-1β),monocyte chemotactic protein-1 (MCP-1) and CB2R (by Western blot) and expression of CD11b in hippocampal tissues (by immunofluorescence).Results There was no significant difference in the total exploring distance in open field test,percentage of freezing time in contextual fear conditioning test or percentage of freezing time in tone-fear conditioning text at each time point among the three groups (P＞0.05).Compared with group C,the percentage of freezing time in contextual fear conditioning test was significantly decreased,and the expression of TNF-α,IL-1β,MCP-1,CD11b and CB2R was up-regulated after surgery in group S (P＜0.05).Compared with group S,the percentage of freezing time in contextual fear conditioning test was significantly increased,and the expression of TNF-α,IL-1β,MCP-1,CD11band CB2R was down-regulated after surgery in group S+J (P＜ 0.05).Conclusion Activation of hippocampal CB2R may be involved in the endogenous protective mechanism of POCD in mice,and the mechanism is related to inhibiting activation of hippocampal microglia and attenuating central inflammatory responses.

Objective:To compare the effects of desflurane and sevoflurane anesthesia on the sleep quality of sleep-deprived mice.Methods:Thirty-two clean-grade healthy male C57BL/6 mice, aged 10 weeks, weighing 20-25 g, were divided into 4 groups ( n=8 each) by the random number table method: control group (C group), sleep deprivation group (SD group), sleep deprivation+ sevoflurane group (SD+ SEV group), and sleep deprivation+ desflurane group (SD+ DES group). In the four groups, EEG-EMG electrodes were implanted for recording EEG and EMG, and sleep deprivation model was developed by the gentle stimulation method with a brush for 12 h (6: 00-18: 00) after 7 days of adaptation. The 6 h after sleep deprivation was divided into 2 time periods: T 1 period (18: 00-20: 00) and T 2 period (20: 00-24: 00). T 1 period In SD group, mice were allowed ad libitum recovery sleep after sleep deprivation. C group and SD group were exposed to 60% oxygen 1.5 L/min. In SD+ DES group and SD+ SEV group, mice were exposed to 6% desflurane and 2.5% sevoflurane, respectively, for 2 h in 60% oxygen 1.5 L/min following sleep deprivation. T 2 period Four groups were allowed ad libitum recovery sleep with the EEG-EMG signal recording. The percentages and number of wakefulness time, rapid eye movement time and non-rapid eye movement time during each time period were calculated using Lunion Data software. Results:Compared with C group, the percentage of non-rapid eye movement time and the percentage of rapid eye movement time were significantly decreased, and the percentage of wakefulness time was increased during 12 h sleep deprivation in SD group, SD+ SEV group and SD+ DES group ( P<0.05). Compared with T 1 period, the percentage of non-rapid eye movement time was significantly increased, and the percentage of wakefulness time and percentage of rapid eye movement time were decreased in T 2 period in SD group ( P<0.05). Compared with SD group, the percentage of non-rapid eye movement time and percentage of rapid eye movement time were significantly decreased, and the percentage of wakefulness time was increased in T 2 period in SD+ SEV group and SD+ DES group ( P<0.05). There was no significant difference in the percentage of non-rapid eye movement, rapid eye movement and wakefulness time in T 2 period between SD+ SEV group and SD+ DES group ( P>0.05). Compared with SD+ SEV group, the number of non-rapid eye movement in T 2 period was significantly reduced in SD+ DES group ( P<0.05). Conclusions:The effect of desflurane anesthesia in improving sleep quality is better than sevoflurane anesthesia in sleep-deprived mice.

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 investigate the effect of CT total tumor perfusion parameters in the identification of benign and malignant lung lesions. Methods 80 patients with pulmonary space-occupying lesions who were treated in our hospital from December 2015 to May 2017 were enrolled. After pathological histological diagnosis, 52 cases of malignant lesions and 28 cases of benign lesions were confirmed. And they all underwent CT whole tumor perfusion scan and perfusion imaging analysis. And the blood flow, blood volume, average transit time and surface permeability were obtained.Some cases of histopathological specimens were treated by monoclonal CD34 staining method, and the microvascularrich areas in the specimens were selected for blood vessel counting. The morphology of blood vessels was observed and microvessel density was obtained. Results The levels of BF, BV, MTT and PS in benign lesions were significantly lower than those in patients with malignant lesions. The best diagnostic threshold, sensitivity and specificity of BF in lung occupying lesions was> 49.12 mL/100 mL, 80.00％ and 52.40％. The best diagnostic threshold, sensitivity and specificity of BV was 6.68 m L/100 mL, 70.00％ and 81.00％. The best diagnostic threshold, sensitivity and specificity of MTT was3.82 m L/100 mL, 97.50 ％ and 52.40％. The best diagnostic threshold, sensitivity and specificity of PS was 7.25 mL/100 mL, 92.50％ and 90.50％. The malignant lesion microvessel density count in the malignant lesions was significantly higher than that of benign lesions, and the difference was statistically significant (P<0.05). Conclusion CT total tumor perfusion parameters have a great auxiliary effect on the diagnosis of benign and malignant lung occupying lesions. Among them, PS and BV have the strongest sensitivity and are worthy of clinical promotion.

Objective:To evaluate the role of activation of vesicular glutamate transporter 2 (VGLUT2) neurons in vagal nodose ganglion in dexmedetomidine-caused bradycardia in mice.Methods:Ninety-six SPF healthy male VGLUT2-cre mice, aged 10 weeks, weighing 20-25 g, were divided into 6 groups ( n=16 each) by the random number table method: normal saline control group (NS group), dexmedetomidine group (Dex group), viral control + chemogenetic control + dexmedetomidine group (eGFP-NS+ Dex group), viral transfection + chemogenetic control + dexmedetomidine group (hM4Di-NS+ Dex group), viral control + chemogenetic inhibition + dexmedetomidine group (eGFP-CNO+ Dex group) and viral transfection + chemogenetic inhibition + dexmedetomidine group (hM4Di-CNO+ Dex group). Dexmedetomidine 100 μg/kg was intraperitoneally injected in Dex group. The equal volume of normal saline was intraperitoneally injected in NS group. AAV2/9-hSyn-DIO-hM4Di-eGFP was injected in the right nodose ganglion in hM4Di-NS+ Dex group and hM4Di-CNO+ Dex group, and AAV2/9-hSyn-DIO-eGFP was injected in the right nodose ganglion in eGFP-NS+ Dex group and eGFP-CNO+ Dex group, allowing the virus expression for 21 days. On the 22nd day after virus injection, clozapine-n-oxide (CNO) 5 mg/kg was intraperitoneally injected in hM4Di-CNO+ Dex group and eGFP-CNO+ Dex group, the equal volume of normal saline was intraperitoneally injected in hM4Di-NS+ Dex group and eGFP-NS+ Dex group, 1 h later the efficacy of CNO reached the peak, and then dexmedetomidine 100 μg/kg was intraperitoneally injected. The respiratory rate, heart rate, SpO 2 and discharge frequency of the right vagal nodose ganglion were synchronously measured by multi-channel electrophysiology in vivo. The expression of phosphorylated extracellular signal-regulated kinase (pERK) and VGLUT2 and co-expression of pERK and VGLUT2 in the right vagal nodose ganglion were detected by immunofluorescence assay. Results:Compared with NS group, the percentage of heart rate variation and neuron firing frequency after administration were significantly increased, and pERK expression was up-regulated in the other five groups ( P<0.05). Compared with Dex group, the percentage of heart rate variation and neuron firing frequency after administration were significantly decreased, and pERK expression was down-regulated in hM4Di-CNO+ Dex group, and no significant change was found in the parameters mentioned above in hM4Di-NS+ Dex group, eGFP-NS+ Dex group and eGFP-CNO+ Dex group ( P>0.05). Compared with hM4Di-CNO+ Dex group, the percentage of heart rate variation and neuron firing frequency after administration were significantly increased, and pERK expression was up-regulated in eGFP-CNO+ Dex group ( P<0.05). There was no significant difference in the percentage of respiratory variation and SpO 2 among the six groups ( P>0.05). The expression of VGLUT2-positive neurons was abundant in nodose ganglia, and the co-expression rate of pERK and VGLUT2 was nearly 90%. The co-expression rate of pERK and VGLUT2 decreased to about 30% after inhibition of VGLUT2 neurons in ganglion. Conclusions:The mechanism by which dexmedetomidine causes bradycardia is associated with activation of VGLUT2 neurons in vagal nodose ganglia in mice.

Objective:To evaluate the effect of propofol on parvalbumin (PV) neurons in the medical prefrontal cortex(mPFC)of rats with social behavior disorders induced by chronic sleep deprivation.Methods:Forty-two SPF male Sprague-Dawley rats, aged 8 weeks, weighing 200-250 g, were divided into 3 groups ( n=14 each) using a random number table method: control group (group Con), chronic sleep deprivation plus natural sleep group (group CSD+ NS), and chronic sleep deprivation plus propofol group (group CSD+ Pro). Sleep deprivation model was established by the modified multiple platform method, the rats were placed in the sleep-deprivation tank for 20 h a day (14: 00-10: 00), and allowed to sleep naturally for 4 h (10: 00-14: 00) a day for 28 consecutive days. Propofol 40 mg/kg was intraperitoneally injected for 28 consecutive days after sleep deprivation in CSD+ Pro group. While the equal volume of 10% fat emulsion was given in Con and CSD+ NS groups. After the end of sleep deprivation, a three-box social experiment was used to detect the social behavior of rats, and the number of the PV positive cells and density of the perineuronal network (PNN) in the mPFC area were measured by immunofluorescence. Results:Compared with group Con, the pertentage of rapid eye movement sleep and sniffing time preference coefficients for the strange rat 1 in the first stage and for the strange rat 2 in the second stage were significantly decreased, and the number of the PV positive cells and density of PNN in the mPFC area were decreased in group CSD+ NS ( P<0.05). Compared with group CSD+ NS, the sniffing time preference coefficients for the strange rat 1 in the first stage and for the strange rat 2 in the second stage were significantly increased, the number of the PV positive cells and density of PNN in the mPFC area were increased( P<0.05), and no significant change was found in the percentage of the rapid eye movement sleep in group CSD+ Pro. Conclusions:Propofol probably increases the number and function of PV neurons in the mPFC and ameliorates sleep deprivation-induced social behavior disorders in sleep-deprived rats.

Objective:To investigate the role of dopamine receptors in the central amygdala (CeA) in reduction of the anxiety level by propofol in a mouse model of post-traumatic stress disorder (PTSD).Methods:Fifty-six SPF healthy adult male C57BL/6 mice, aged 10 weeks, weighing 20-25 g, were divided into 7 groups ( n=8 each) using a random number table method: control group (C group), PTSD group (P group), PTSD+ propofol group (PP group), PTSD+ fat emulsion group (PF group), PTSD+ propofol+ normal saline group (PPN group), PTSD+ propofol+ dopamine receptor D1 (DRD1) antagonist group (PP+ DRD1-Ant group), and PTSD+ propofol+ DRD2 antagonist group (PP+ DRD2-Ant group). The PTSD model was developed by continuous plantar electric shock for 3 days. Propofol 120 mg/kg was intraperitoneally injected after successful establishment of the model in PP group, and the equal volume of fat emulsion was intraperitoneally injected in PF group. In PPN group, PP+ DRD1-Ant group and PP+ DRD2-Ant group, the equal volume of normal saline, DRD1 antagonist hydrochloride and DRD2 antagonist eticlopride hydrochloride were injected in bilateral CeA regions, respectively, 30 min later the efficacy of drugs reached the peak, and then propofol 120 mg/kg was intraperitoneally injected. The anxiety levels were measured at 4 h (T 1) and day 3 after propofol injection (T 2) by the open field test and elevated cross maze test. Results:Compared with C group, the time spent entering the open and central areas was significantly shortened at T 1, 2, and the number of entering the open and central areas was decreased at T 1, 2 in P group ( P<0.001). Compared with P group, the time spent entering the open and central areas was significantly prolonged at T 1, the number of entering the open and central areas was increased at T 1 ( P<0.001), and no significant change was found at T 2 in PP group ( P>0.05), and no significant change was found in the aforementioned parameters at T 1, 2 in PF group ( P>0.05). Compared with PPN group, the time spent entering the open and central areas was significantly shortened at T 1, and the number of entering the open and central areas was decreased at T 1 in PP+ DRD2-Ant group ( P<0.001), and no significant change was found at T 1 in PP+ DRD1-Ant group ( P>0.05). Conclusions:Activation of DRD2 in the CeA is involved in the process by which propofol reduces the anxiety level of mice with PTSD.