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Objective To evaluate the role of group Ⅱ metabotropic glutamate receptors (mGluRs) in cognitive decline caused by multiple administrations of ketamine in mice and the relationship with hippocampal glycogen synthase kinase-3 beta (GSK-3β) expression.Methods Forty-five SPF healthy female C57BL/6 mice,aged 6-8 weeks,weighing 20-30 g,were randomized into 3 groups (n=15 each) using a random number table method:control group (group C),ketamine group (group K) and mGluR agonist LY354740 group (group L+K).In K and L+K groups,ketamine 30 mg/kg was intraperitoneally injected three times a day at an 30-min interval for 14 consecutive days.LY354740 was intraperitoneally injected at 30 min before the first injection of ketamine in group L+K.The equal volume of normal saline was given instead in group C.Morris water maze test was performed the day after the last administration.The mice were then sacrificed,and hippocampi were harvested to determine the expression of GSK3β,NR2A and postsynaptic density protein 95 (PSD95) by Western blot.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,the frequency of crossing the original platform was decreased,the expression of GSK3β3 and NR2A was up-regulated,and the expression of PSD95 was down-regulated in group K (P＜0.05),and no significant change was found in the parameters mentioned above in group L+K (P＞0.05).Compared with group K,the escape latency was significantly shortened,the time of staying at the original platform quadrant was prolonged,the frequency of crossing the original platform was increased,the expression of GSK3β and NR2A was down-regulated,and the expression of PSD95 was up-regulated in group L+K (P＜0.05).Conclusion Group Ⅱ mGluRs are involved in the process of cognitive decline caused by multiple administrations of ketamine in mice,which is associated with up-regulated expression of hippocampal GSK-3β.
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Objective To explore the application of bundle management in enhanced recovery after surgery (ERAS) on liver transplantation.Methods The multidisciplinary team of West China Hospital of Sichuan University discussed the program of ERAS on liver transplantation in 2016,and implemented this program in July 2016.A retrospective analysis was made on 220 liver transplant patients who were admitted to West China Hospital of Sichuan University in the period from Jan.2015 to Mar.2017.According to the inclusion and exclusion criteria,there were 104 patients in traditional group and 92 patients in ERAS group.The clinical indicators during and after surgery were compared between the two groups,and the applied value of ERAS on liver transplantation was analyzed.Results As compared with the traditional group,the patients in ERAS group had advantages in operative time,blood loss,postoperative stay in intensive care unit (ICU),transfusion volume in ICU,endotracheal intubation time and total hospitalization time,with significant difference between the two groups (P＜0.05).Meanwhile readmission and mortality rate after one month in ERAS group was not increased.Conclusion The bundle management in ERAS on liver transplantation of West China Hospital of Sichuan University can improve the prognosis of liver transplant patients.
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Objective To evaluate the role of nuclear factor kappa B (NF-κB) in cognitive decline in aged mice with sepsis.Methods Forty-five SPF healthy aged female C57BL/6 mice,aged 10-12 months,weighing 20-30 g,were assigned into 3 groups (n=15 each) using a random number table:control group (group C),sepsis group (group Sep) and NF-κB selective inhibitor pyrrolidine dithiocarbamate (PDTC) group (group PDTC).Lipopolysaccharide 250 μg/kg was injected intraperitoneally once a day for 7 consecutive days in Sep and PDTC groups,and in addition PDTC 50 mg/kg was injected intraperitoneally at 30 min before lipopolysaccharide injection once a day for 7 consecutive days in group PDTC.The equal volume of normal saline was given in group C.Five mice in each group were sacrificed at 2 h after the last administration,cardiac puncture was performed and blood samples were collected,and then the mice were sacrificed and hippocampi were harvested for determination of the levels of tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and IL-6 in plasma and hippocampal tissues using enzyme-linked immunosorbent assay.Cognitive function was assessed using open field,elevated plus maze and Morris water maze tests at 24 h after the last administration in the other mice left in each group.Results Compared with group C,the levels of TNF-α,IL-1β and IL-6 in plasma and hippocampal tissues were significantly increased,the time of movement at the central region was shortened,the percentage of time spent in the open arms and number of entries into the open and closed arms were decreased,the escape latency was prolonged,the time of staying at the original platform quadrant was shortened,and the frequency of crossing the platform was decreased in group Sep (P＜0.05).Compared with group Sep,the levels of TNF-α,IL-1β5 and IL-6 in plasma and hippocampal tissues were significantly decreased,the time of movement at the central region was prolonged,the percentage of time spent in the open arms and number of entries into the open and closed arms were increased,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in group PDTC (P＜0.05).Conclusion NF-κB is involved in cognitive decline in aged mice with sepsis.
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Objective To evaluate the efficacy of pressure support ventilation ( PSV ) in the infants undergoing laparoscopic hernia repair under sevoflurane anesthesia. Methods Thirty ASA physical statusⅠpediatric children, aged 9 months-1 yr, weighing 8.0-11.5 kg, undergoing elective laparoscopic hernia repair, were randomly assigned into 3 groups ( n=10 each) using a random number table: pressure control ventilation ( PCV) used for muscle relaxants in combination with low?concentration sevoflurane group ( group PCV1 ) , PCV used for high?concentration sevoflurane group ( group PCV2 ) , and PSV used for low?concentration sevoflurane group ( group PSV) . Anesthesia was induced with inhalation of 4%-6%sevoflurane and iv fentanyl 2 μg∕kg and succinylcholine 1.5 mg∕kg. The pediatric children were endotracheally intubated and mechanically ventilated. In PCV1 and PCV2 groups, PCV was used during operation. In group PSV, PCV was used first after intubation, and then PSV was applied after spontaneous breathing recovered. Anesthesia was maintained as follows: in group PCV1 , the end?tidal concentration of sevoflurane was maintained at 2.5% - 3.0%, and cisatracurium besylate 0.1 mg∕kg was injected intermittently as required; in group PCV1 , the end?tidal concentration of sevoflurane was maintained at 3.5%-4.0%; in group PSV, the end?tidal concentration of sevoflurane was maintained at 2.5%-3.0%, and succinylcholine 1.0 mg∕kg was injected intravenously before pneumoperitoneum. Narcotrend index value was maintained at 50-60 in PCV1 and PSV groups, or at 37-45 in PCV2 group. Heart rate ( HR) and mean arterial pressure (MAP) were recorded before induction of anesthesia (baseline), at the beginning of pneumoperitoneum, at 5 and 10 min of pneumoperitoneum, at the end of pneumoperitoneum, at the end of operation and immediately after extubation. The time interval from the end of surgery to extubation was recorded. Results Pulse oxygen saturation was 100% during anesthesia, and>95% during recovery from anesthesia in the three groups. Compared with the baseline value, HR was significantly faster, and MAP was increased during extubation in PCV1 and PCV2 groups, and no significant change was found in HR and MAP at each time point in group PSV. The time interval from the end of surgery to extubation was 30.3± 5.4, 18.4±4.3 and (4.1±1.2) min in PCV1, PCV2 and PSV groups, respectively. Compared with PCV1 and PCV2 groups, the time interval from the end of surgery to extubation was significantly shortened in group PSV. Conclusion When PSV is applied in the infants undergoing laparoscopic hernia repair under sevoflurane anesthesia, it can provide adequate ventilation, recovery from anesthesia is rapid, and no cardiovascular responses occur during extubation.
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Objective To evaluate the effects of isoflurane anesthesia on inflammatory responses and long-term cognitive function in the hippocampi of neonatal rats .Methods Forty-six Sprague-Dawley rats of both sexes , aged 7 days ,weighing 12-17 g ,were randomly divided into 2 groups (n=23 each):control group (group C) and isoflurane anesthesia group (group I ) .In group I ,the rats were exposed to 2.5% isoflurane for 3 min to induce anesthesia and then exposed to 1.5% isoflurane for 4 h to maintain anesthesia ,while in group C the rats were only exposed to air for 4 h .Eight rats in each group were sacrificed and hippocampi were removed for determination of the levels of interleukin-6 ,interleukin-1βand tumor necrosis factor-α.Open field and Morris water maze tests were carried out three weeks later in the left rats .Results Compared with group C ,the escape latency was significantly prolonged ,the time of staying at the central region was shortened ,the time of staying at the border region was prolonged ,the total distance was reduced , and the contents of interleukin-1β and tumor necrosis factor-αwere increased in group I ( P<0.05) .Conclusion Isoflurane anesthesia results in decreased cognitive function ,which may be related to promotion of inflammatory responses in the hippocampi of neonatal rats .
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Objective To evaluate the effects of ulinastatin pretreatment on cognitive dysfunction induced by chronic exposure to ketamine in immature mice.Methods Thirty-six healthy male C57BL/6 mice,aged 21 days,weighing 20-30 g,were randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),ketamine group (group K),and ulinastatin pretreatment group (group U).In K and U groups,ketamine 30 mg/kg was injected intraperitoneally three times a day at 30-minute intervals for 21 consecutive days,while in group U,ulinastatin 50 000 U/kg was injected intraperitoneally at 30 min before the first injection of ketamine everyday.Cognitive function was assessed using Morris water maze and open field tests at 24 h after the last administration of ketamine.Mice in each group were sacrificed immediately after the end of the tests and hippocampi were harvested to determine the contents of interleukin-6 (IL-6),IL-1 and tumor necrosis factor-α (TNF-α) using ELISA.Results Compared with group C,the escape latency was significantly prolonged,the time spent in the original platform and in the central area for the open field was shortened,the frequency of crossing the original platform was decreased,and the contents of IL-1,IL-6,and TNF-α were increased in group K (P ＜ 0.05),while there were no significant differences in the indexes mentioned above in group U (P ＞ 0.05).Compared with group K,the escape latency was significantly shortened,the time spent in the original platform and in the central area for the open field was prolonged,the frequency of crossing the original platform was increased,and the contents of IL-1,IL-6,and TNF-α were decreased in group U (P ＜ 0.05).Conclusion Ulinastatin pretreatment can improve cognitive dysfunction induced by chronic exposure to ketamine in immature mice,and inhibition of inflammatory responses in hippocampi may be involved in the mechanism.
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Objective To investigate the effects of isoflurane anesthesia on proliferation and differentiation of neural stem cells (NSCs) in the dentate gyrus of neonatal rats.Methods Ten SD rats,aged 7 days,weighing 16-20 g,were randomly divided into 2 groups ( n =5 each):control group (group C) and isoflurane group (group 1) .The rats in group Ⅰ inhaled 2.5％ isoflurane for 3 min for induction and then anesthesia was maintained with 1.5 ％ isoflurane for 4 h,while the rats in group C only breathed the room air for 4 h.Bromodeoxyuridine (BrdU) 100 mg/kg was injected intraperitoneally right before the induction and after the end of administration to label NSCs and their progeny in the dentate gyrus.At 24 h after the 2nd administration of BrdU,double immunofluorescence for BrdU and NeuroD (a marker of neuroblasts and immature neurons) was used to assess NSC proliferation and neuronal differentiation.Results Compared with group C,the number of BrdU+ cells in group Ⅰ was significantly decreased,whereas the fraction of NeuroD+/BrdU+ differentiated cells was increased in the dentate gyrus( P ＜ 0.05 or 0.01 ).Conclusion Isoflurane anesthesia suppresses the proliferation of NSCs and induces neuronal differentiation of NSCs in the dentate gyrus of neonatal rats.
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ObjectiveTo investigate the effects of isoflurane anesthesia on the expression of brain-derived neurotrophic factor (BDNF) and phosphorylated extracellular signal-regulated kinase (p-ERK) in neonatal rat hippocampus.MethodsForty-eight SD rats of both sexes,aged 7 days,weighing 12-17 g,were randomly divided into 2 groups ( n =24 each):control group (group C) and isoflurane anesthesia group (group Ⅰ).In group Ⅰ,the rats were exposed to2.5％ isotlurane for 3 min and then 1.5％ isoflurane was inhaled for 4 h,while in group C the rats were exposed to air for4 h.Arterial blood samples were collected immediately after anesthesia for blood gas analysis and for determination of the blood glucose concentration.Five rats in each group were sacrificed at 0,6,24 and 48 h after anesthesia (T1-4) and hippocanpi were removed for determination of the expression of potassiumchloride cotransporter 2 (KCC2),potassium-chloride cotrmsporter 1 (NKCC1),BDNF and p-ERK by Western blot.NKCC1/KCC2 ratio was calculated.ResultsAcid-base imbalance,hypoxemia and glycopenia were not found immediately after anesthesia in both groups.Compared with group C,KCC2 expression was significantly down-regulated and NKCC1/KCC2 ratio was increased at T3 and T4,and the expression of BDNF and p-ERK was dewn-regnlated at T1 and T2 in group Ⅰ (P＜0.05).There was no significant difference in NKCCI expression at each time point between groups Ⅰ and C ( P ＞ 0.05 )、ConclusionIsoflurane anesthesia delays the neuronal development in neonatal rat hippocampus through down-regulating the expression of BDNF and p-ERK.
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Objective To investigate the effect of precondition with Toll-like receptor 4 monoclonal antibody (TLR4mAb) on lipopolysaccharide (LPS) -induced acute lung injury in mice.Methods A total of 45 male BALB/c mice were randomly divided into 3 groups:the control group ( group C),the sepsis group (group S) and the pretreatment group (group P).Mice in the group P and group S were injected intraperitoneally with LPS ( 10 mg/kg) to produce acute lung injury models.Mice in the group P was injected intraperitoneally with TLR4mAb (5 μg/g) 1 h before the injection of LPS.Expression of TLR4mRNA in lung tissue,expression of TNF-α and IL-6 in serum,water content of lung,and the pathomorphologic changes of lung were detected after 6 h,12 h and 24 h.One-way ANOVA was used for inter-group comparison and two-way ANOVA was used for intra-group comparison.Results Compared to group C,water content significantly increased at 12 h and 24 h in group S and group P; compared to group S,water content significantly decreased in group P at 12 h and 24 h.Compared to group C,the expression of IL-6 and TNF-α significantly increased in group S and group P at 6 h,12 h and 24 h; compared to group S,the expression of IL-6 and TNF-α significantly decreased at 6 h,12 h and 24 h in group P.Compared to group C,the expression of TLR4 mRNA increased significantly in group S and group P at 6 h,12 h and 24 h; compared to group S,the expression of TLR4 mRNA decreased significantly in group P at6 h,12 h and 24 h.Compared to group S,pathological damage of the lung was improved significantly in group P.Conclusions Precondition with TLR4mAb can attenuate LPS-induced acute lung injury,suppress the expression of inflammatory factors.Regulation of TLR4 pathway may be a promising therapeutic strategy for ALI.
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Objective To investigate the effects of propofol on the development of spatial learning and memory and neuron proliferation of neonatal rats at different doses. Methods 60 neonatal rats were divided into four groups among per litter by using a randomized block design. Three different doses of propofol group were induced with propofol 10 mg/kg( group P10) ,50 mg/kg( group P50) or 50 mg/kg twice( group P50D) by subcutaneous injection respectively. Neuron proliferation at dentate gyrus was detected by using BrdU marker 3 days later.Morris water maze test was carried out on postnatal day 28. Escape latency,time in probe quadrant were recorded.Results Compared to the control group,neuron marked with BrdU at dentate gyrus in group P50D was significantly decreased( (840±76) vs (225 ±66), P＜0.05) ,group P10 was significantly increased( (840 ±76) vs ( 1225± 154), P＜0.05). Compared to the control group,latency of group P50D was significantly increased( ( 15.12 ±3.43 ) s vs (42.68 ± 6. 18 ) s, P ＜ 0. 05 ), time in probe quadrant of group P50D were significantly decreased ( ( 55.66 ± 8.57 ) s vs (32. 18 ± 5. 38 ) s, P＜ 0. 05 ). Compared to the control group, there was no significant difference between group P50 and group P10. Conclusion Propofol given to seven-day-old rats with 50 mg/kg twice by subcutaneous injection suppresses neuron proliferation and impairs development of memory and learning in neonatal rats,but propofol given with 10 mg/kg once promotes neuron proliferation.
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Objective To observe the pain behavioral performance of rats that different sensory nerve fibers were transected,and examine the expression of brain-derived neurotrophic factor(BDNF) in these models.Methods Twenty-four rats were divided into three groups according to random number table method:SUR group,GS group and SHAM group, which received sural nerve transection, gastrocnemius-soleus nerve transection or sham operation respectively.There were 8 rats in every group.The expression of BDNF in the lumbar 5 DRG and spinal dorsal horn were detected,and the types of damaged cells were also observed.Results In GS group, 50% paw-withdrawal thresholds were significantly decreased on the ipsilateral hind paw compared with baseline and with those in SHAM group,and the paw-withdrawal durations in response to the thermal stimulus increased significantly (P＜0.01 ＝.In contrast, no change was found in SUR group(P＞0.05 ).The expression of BDNF in the lumbar 5 DRG ( (37.87 ± 4.23 ) % ) and spinal dorsal horn ( (21.9 ± 3.1 ) % ) was significantly higher in GS group than in SHAM group( ( 17.31 ± 2.12 ) %, ( 12.6 ± 1.3 ) % ), and no significant difference was found between SUR and SHAM groups(P＞0.05 ).FG opposite cells which also expressed BDNF in GS group were more than those in SUR group ( (47.7 ± 1.8) % and (26.7 ± 2.3 ) % ) (P ＜ 0.01 ＝.The percentage in N200 and FG double positive cells to N200 positive cells in GS group was significantly increased in GS group than those in SUR group ( (47.7 ±1.8 ) %, (26.7 ± 2.3 ) % ) (P ＜ 0.01 ＝.Conclusion The data suggest that injury of the sensory nerve innervating skin does not produce hyperalgesia, but injury of the sensory nerve innervating muscle does.Different kinds of neuron were damaged and the differences of BDNF expression is essential for this difference.
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Objective To investigate the effects of different concentrations of isoflurane on the caspase-3 expression in the hippocampus and S100β level of plasma in fetal rats. Methods 18 pregnant rats at gestational day 21 were divided into control group, 1. 3% isoflurane group,3% isoflurane group. Rats in the control group spontaneously breathed 100% oxygen for 1 h. Rats in the treatment groups breathed 1.3% or 3% isoflurane in 100% oxygen through an endotracheal tube, with mechanical ventilation for 1 h. Rat pups were delivered by cesarean section 6 h after treatment, and fetal blood was sampled from the left ventricle of each fetal heart and evaluated for S100β. Fetal brains were then evaluated for apoptosis, using caspase-3 immunohistochemistry in the CA1 region of the hippocampus. Results Compared to the control group ((1. 48 ± 0. 08) μg/L) and the 1. 3% isoflurane group( (1.53 ±0. 12)μg/L) ,the 3% isoflurane group showed significantly higher level of S100β( (3. 12 ±0. 15) μg/L, P＜0.05) . There was no differences in densities of caspase-3-positive cells between the control ((33 ±4) cell/mm ) and 1.3% isoflurane groups((31 ±5)cell/mm2). Compared to 1.3% isoflurane,isoflurane at a concentration of 3%((75 ± 7) cell/mm2, P＜0.05) for lh increased neurodegeneration in the hippocampal CA1 area in the developing brain of fetal rats. Conclusion Isoflurane can dose-dependently induce brain damage. Isoflurane at a concentration of 3% for lh can induce apoptosis in the hippocampal CA1 area and increase S100β levels of fetal rats.
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Objective To evaluate the role of protein kinase C (PKC) in reduction of hypoxia-reoxygenation (H/R) injury by hypoxic preconditioning or norepinephrine preconditioning in cultured neonatal rat cardiomyocytes. Methods Primary cultured neonatal rat cardiomyocytes were randomly divided into 6 groups (n = 25 each): control group (group Ⅰ), H/R group (group Ⅱ), hypoxia preconditioning group (group Ⅲ), norepinephrine preconditioning group (group Ⅳ), H7 + hypoxia preconditioning group (group Ⅴ) and H7 + norepinephrine preconditioning group (group Ⅵ). In group Ⅱ , the cardiomyocytes were exposed to 3 h of hypoxia followed by 1 h of reoxygenation. In group Ⅲ, the cells were subjected to 20 min of hypoxia followed by 20 min of reoxygenation before H/R. Norepinephrine was added to the culture medium with a final concentration of 10- 7 mol/L,and then the cells were cultured for 30 min before H/R in group Ⅳ. H7 was added to the culture medium with a final concentration of 5 × 10-5 mol/L, the cells were then cultured for 10 min, and the following procedures before H/R were the same as thase described in group Ⅴ . H7 was added to the culture medium with a final concentration of 5 × 10-5 mol/L, the cells were then cultured for 10 min, and the following procedures were the same as those described in group Ⅵ. The cell survival rate, the activities of LDH and CK in the supernatant, and the content of MDA and activity of SOD in cardiomyocytes were determined. Results The cell survival rate and activity of SOD were significantly lower, while the LDH and CK activities and MDA content higher in group Ⅱ than in group Ⅰ ,in group Ⅴ than in group Ⅲ, and in group Ⅵ than in group Ⅳ (P ＜ 0.01). The cell survival rate and activity of SOD were significantly increased, while the LDH and CK activities and MDA content decreased in group Ⅲ and Ⅳ compared with group Ⅱ (P＜0.01).Conclusion The activiation of PKC is involved in the reduction of H/R injury by hypoxic preconditioning or norepinephrine preconditioning in cultured neonatal rat cardiomyocytes.
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Objective It has been shown that strong acute stress or long-term chronic stress significantly affects learning and memory. The aim of this study was to investigate the effects of chronic pain on learning and memory and morphological structure of hippocampus in neonatal rats.Methods Sixty SD rats aged 7 days were randomly divided into two groups : (A) chronic pain group ( n = 30) in which 0.5% formalin 0.1 ml was injected subcutaneously into plantar region of hind paw every day for two weeks and (B) control group (n = 30) in which the plantar region of hind paw was touched with cotton-tipped swab every day for 2 weeks instead of subcutaneous injection of formalin. Morris water maze performance was used to test learning and memory. The number of granule neurons in dentate gyrus and pyramidal neurons in CA3 were counted. Results The mean latency period in the Morris water maze intelligence test was significantly longer in chronic pain group than that in control group ( P
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Objective To evaluate the feasibility and safety of using continuous spinal anesthesia (CSA) in high risk elderly patients undergoing lower abdominal or lower extremity surgery. Methods Sixty-four ASA III or IV patients aged 70-101 yr weighing 38-55 kg undergoing lower abdominal or lower extremity surgery were randomized to receive epidural anesthesia (EA) (n = 32) or CSA ( n = 32). The patients were complicated with cerebral embolism and/or hypertension, coronary artery disease and/or COPD and/or diabetes mellitus. The patients were unpremedicated. EA was performed at I2.3 or L1.2. A test dose of 2 ml of 2% lidocaine was given. When no signs of spinal block was observed, 1 % ropivacaine was given in small increments until the block height reached T6-8 . CSA was performed at L3,4 using Spinocath (B. Braun). A 2-cm catheter segment was left in subarachnoid space. 0.5% bupivacaine was given in 0.5 ml increments every 3 min until satisfactory block level was reached. The onset time of anesthesia was recorded. The degree of motor block was assessed using modified Bromage scale. Arterial blood samples were obtained before anesthesia (T0 , baseline), when satisfactory block level was reached ( T1), 1 h after skin incision (T2) and at the end of operation (T3) for determination of lactate concentration. The amount of ephedrine and innovar ( droperidol-fentanyl mixture) used during operation and the recovery of motor function of lower extremities were also recorded. Results The demographic data including sex, age, height, body weight, ASA physical status and types of operation were not significantly different between the two groups. BP and HR were significantly decreased after anesthesia as compared to the baseline values at T0 in group EA, whereas in CSA group there were no significant changes in BP and HR after anesthesia. Significantly more patients received ephedrine in EA group (98% ) than in CSA group (15%) and the mean dose of ephedrine was significantly higher in EA group [(34.5?3.1) mg] than in CSA group [(4.3?0.5) mg ]. The onset of block was significantly faster in CSA group than in EA group. Motor blockade was less intense in EA group as assessed by modified Bromage scale. Analgesia was more satisfactory in CSA group and less patients received innovar during operation in CSA group (20% ) than in EA group (51% ). The recovery of motor function of lower extremities was faster in CSA group than in EA group. Blood lactate was significantly higher at T1 , T2 , T3 in EA group than in CSA group. No late complications related to CSA was observed. Conclusion CSA is safe and effective and provides better analgesia with better hemodynamic stability and faster onset of block than EA in elderly patients.