Objective To evaluate the effect of isoflurane on the apoptosis of SH-SYSY cells transfected with APPsw gene and the role of inositol 1,4,5-triphosphate (IP3) recepters.Methods The SH-SYSY ceils transfected with APPsw gene were seeded in culture flasks with the density of 1.2 × 104/cm2.The cells were randomly divided into 4 groups (n =6 each):control group (group C),IP3 receptor antagonist group (group Ⅹ),isoflurane group (group Ⅰ) and isoflurane + IP3 receptor antagonist group (group Ⅰ + Ⅹ).After the cells were cultured for 24 h and attached to the wall,the cells were cultured routinely in group C,and Xestospongin C 100 nmol/L (IP3 receptor antagonist) was added to DMEM culture medium in groups X and Ⅰ + X,and 30 min later the cells were exposed to 1.2 ％ sevoflurane for 8 h in groups Ⅰ and Ⅰ + X.The cells were collected for examination of the ultrastructure and for determination of cell apoptosis,intracellular free calcium ion concentration [Ca2 +] i (by flow cytometry) and expression of IP3 receptor protein (by Western blot).The apoptosis rate was calculated.Results Compared with group C,there was no significant change in the apoptosis rate,[Ca2 +]i or IP3 receptor protein expression in group Ⅹ (P ＞ 0.05),while the cell apoptosis rate and [Ca2 +] i were significantly increased and IP3 receptor protein expression was up-regulated in groups I and Ⅰ + Ⅹ (P ＜ 0.05 or 0.01).Compared with group Ⅰ,cell apoptosis rate and [Ca2+]i were significantly decreased and IP3 receptor protein expression was down-regulated in group Ⅰ + Ⅹ (P ＜ 0.01).The pathological changes of the cells happened in groups Ⅰ and Ⅰ + Ⅹ,and the pathological changes were severer in group Ⅰ than in group Ⅰ + Ⅹ.Conclusion Isoflurane can induce apoptosis of SH-SY5Y cells transfected with APPsw gene through increasing [Ca2+]i and up-regulating IP3 receptor protein expression.

BACKGROUND:Sleep deprivation, resulting in a series of physiological and psychological reactions, is a common phenomenon in our modem society. Recently, many physical and medical therapies are on their way to eliminate the negative influence the sleep deprivation exerted on our human beings, and a large number of cigarette smokers believe that cigarette smoke can obviously improve their abnormal behavioral performance and negative emotional fluctuation caused by sleep deprivation in short terms.However, there are few researches but many disputes when it comes to effect of passive smoking on the behavior and emotion of rats on the occasion of acute sleep deprivation.OBJECTIVE: To investigate the effect of passive smoking (PS) on the behavior and emotion of rats with acute sleep deprivation (SD).DESIGN: Completed randomized and controlled study.SETTING: Basic Medicine Experiment Center, School of Basic Medicine;Department of Radiation Medicine, School of Preventive Medicine, Fourth Military Medical University of Chinese PLA.MATERIALS: The experiment was carried out at Experiment Center of Basic Medicine, Fourth Military Medical University of Chinese PLA from April to June 2004. Totally 49 healthy adult male Sprague-Dawley rats,being 3 months old, weighting (180±15) g, were randomly divided into 5groups: sleep deprivation + 24-hour passive smoking group (n=8), 24-hour sleep deprivation group (n=8), sleep deprivation + 54-hour passive smoking group (n=8), 54-hour sleep deprivation group (n=8), and blank control group (n=7).METHODS: ① Rats in sleep deprivation + 24-hour passive smoking group, 24-hour sleep deprivation group, sleep deprivation + 54-hour passive smoking group and 54-hour sleep deprivation group were performed with sleep deprivation with flower pot method for 24 and 54 hours, and rats in sleep deprivation + 24-hour passive smoking group and sleep deprivation + 54-hour passive smoking group were treated with passive smoking at the same time at a frequency of 12 times/day, 120 minute/time, 3cigarettes were lighted at the same time and kept into ashes. ② Broad field test was conducted to determine the behavioral and emotional changes of the rats. We trailed their activities by video recorder for 5 minutes each time. Then we presented the image information to computer process and finally got the total distance and the average distance from the center of the broad field for each rat during the 5-minite test. Meanwhile, we also observed the emotional variety and excitability of the rats. The nearer the average distance from center was, the more the emotional behavior and the greater excitability were; and the longer the total distance was, the greater the excitability was. ③ All the data were expressed with Mean ± SD, the method of non-parametric rank sum test was adopte d for comparing among groups, P ＜ 0.05 was regarded as statistical significance.MAIN OUTCOME MEASURES: Comparisons between DC and TD of rats in each group, and changes of behavior and emotion of rats in each group.RESULTS: Data of all the 49 rats was entered the final data analysis without any loss. ① Observation of emotional stage: Compared with sleep deprivation + 54-hour passive smoking group, rats in sleep deprivation+24-hour passive smoking group appeared more quiet and indifferent, and presented lower excitability, less sensitivity to outside stimulus and less aggressive behaviors towards other rats, and the same as 24-hour sleep deprivation group, compared with 54-hour sleep deprivation group. ② Results from the open field test: DC of 24-hour sleep deprivation group was significantly smaller than that of blank control group and sleep deprivation + 24-hour passive smoking group [(53.93±1.83, 58.21±4.45, 58.11±1.62) cm, (P＜ 0.01-0.05)]. DC of 54-hour sleep deprivation group was significantly bigger than that of 24-hour sleep deprivation group [(61.53±3.02, 58.11±1.62) cm, (P＜ 0.01)]; TD of 24-hour sleep deprivation group was significantly bigger than that of sleep deprivation + 24-hour passive smoking group [(3310.45±1 445.97, 1 818.20±733.25, 2 338.15±694.70) cm, (P ＜ 0.01-0.05)], and that of 54-hour sleep deprivation group was bigger than that of sleep deprivation + 54-hour passive smoking group andsmaller than that of 24-hour sleep deprivation group [(2 410.70t548.64, 1 473.50±945.89, 3 310.45±1 445.97) cm, (P＜0.05)].CONCLUSION: With the prolongation of sleep deprivation time, behavior and emotional reaction of rats are shown from exciting to inhibiting, however, passive smoking can inhibit behavior and emotional state of rats during the whole sleep deprivation.

Objective To evaluate the effects of sevoflurane preconditioning on postoperative cognitive dysfunction in aged rats.Methods One hundred and twenty healthy male Sprague-Dawley rats,aged 18 months,weighing 400-450 g,were randomly divided into 3 groups (n =40 each) using a random number table:control group (group C),operation group (group O),and sevoflurane preconditioning group (group Sev).In group C,the rats were only anesthetized with pentobarbital sodium and did not undergo operation.In group O,the rats were anesthetized with pentobarbital sodium and underwent 30 min of exploratory laparotomy.In group Sev,the rats inhaled 2.4％ sevoflurane for 30 min and then inhaled air for 30 min,and the other procedures were similar to those previously described in group O.At 30 min before operation and on 1st,3rd,5th and 7th days after operation,Morris water maze test was performed to record the escape latency,time of staying at the original platform quadrant and frequency of crossing the original platform.At 30 min before operation and on 1st and 7th days after operation,10 rats in each group were sacrificed and hippocampi were isolated to detect the apoptotic rate and intracellular [Ca2 +] i (using flow cytometry) and the ultrastructure of hippocampal neurons was observed with transmission electron microscope.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,and the apoptotic rate and intracellular [Ca2 +] i were increased after operation in O and Sev groups (P ＜0.05).Compared with group O,the escape latency was significantly shorten,the time of staying at the original platform quadrant was prolonged,the frequency of crossing the original platform was increased,and the apoptotic rate and intracellular [Ca2+]i were decreased after operation in group Sev (P ＜ 0.05).Microscopic examination showed no abnormality in the ultrastructure of hippocampal neurons in group C,and the pathological changes of the ultrastructure of hippocampal neurons were obvious in group O,and were significantly attenuated in group Sev.Conclusion 2.4％ sevoflurane preconditioning can reduce postoperative cognitive dysfunction in aged rats,and regulation of imbalance of calcium homeostasis and reduction of cell apoptosis are involved in the mechanism.

Objective To evaluate the effects of the prone position on pulmonary gas exchange during mechanical ventilation under general anesthesia.Methods Thirty patients scheduled for elective spine surgery in the prone position under general anesthesia (group prone,n =30),30 patients scheduled for elective spine surgery in the supine position under general anesthesia (group supine,n=30),aged 30-64 yr,with body mass index of 19-30 kg/m2,of ASA physical status Ⅰ or Ⅱ,were enrolled in the study.After induction of general anesthesia,the patients were mechanically ventilated.Anesthesia was maintained with total intravenous anesthesia.At 10 min before pre-oxygenation (T0),10 min after intubation (immediately after the patients were moved from the supine to the prone position) (T1),45 and 90 min after intubation (T2,3),5 min before extubation (immediately before supine position to the prone position) (T4),and 15 min after extubation (T5),arterial blood samples were taken for blood gas analysis,and PaO2 and PaCO2 were recorded.Alveolar-arterial oxygen difference (A-aDO2) was calculated.Digital radiography was performed and the changes of the lung were observed.Results Compared with supine group,PaO2 was significantly increased and A-aDO2 was decreased at T1-4 in prone group.There was no significant difference in PaCO2,and PaO2 and A-aDO2 at T0 and T5 between the two groups.The results of digital radiography showed no atelectasis at different time points in either group.Conclusion Pulmonary gas exchange in the prone position is superior to that in the supine position during mechanical ventilationunder general anesthesia.

Objective: To identify barriers of evidence-based practice in venous blood specimen collection. Methods: Totally 12 nurses from Zhongshan Hospital Fudan University participated in the non-structure interviews. Data were analyzed by category analysis. Results: There were 8 barriers, including standard operation procedure of blood specimen collection was not updated in time, lacking of education and professional support for evidence-based nursing, lacking of effective superintendence, rising economic cost, deficiency of technology and equipment, sufficient nurses, deficiency of evidence-based practice environment. Conclusions Establishing and implementing an evidence-based standard operation procedure, strengthening personnel education, looking for professional support, developing link management, reasonable allocation of human resources and flexible scheduling, creating an evidence-based practice environment, seeking technical and equipment support and evaluating evidence to control costs are benefit to promote the best evidence of venous blood collection to achieve clinical transformation.

Objective:To evaluate the role of RhoA/ROCK2 signaling pathway in multiple exposures to sevoflurane-induced long-term cognitive impairment in neonatal rats.Methods:Sixty SPF healthy neonatal Sprague-Dawley rats of either sex, aged 6 days, weighing 12-20 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), multiple exposures to sevoflurane group (group S) and RhoA/ROCK2 signaling pathway inhibitor Y-27632 group (group Y). Group S and group Y inhaled 3% sevoflurane for 2 h at days 6, 7 and 8 after birth.In group Y, Y-27632 5 mg/kg was intraperitoneally injected before sevoflurane anesthesia.The spontaneous activity was evaluated by open field test on day 35 after birth.The cognitive function was detected by Morris water maze test at day 36 after birth.The rats were sacrificed after Morris water maze test, and the hippocampal tissues were isolated for determination of the apoptosis rate of hippocampal neurons and cytoplasmic calcium concentration ([Ca 2+ ] i) (by flow cytometry) and expression of phosphorylated RhoA (p-RhoA), ROCK2 and cleaved-caspase-3 (by Western blot) and for microscopic examination of the ultrastructure of hippocampal neurons (with a transmission electron microscope). Results:There was no significant difference in movement speed, distance and time of stay in the open field center in the open field test among the three groups ( P>0.05). Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the apoptosis rate of hippocampal neurons and [Ca 2+ ] i were increased, the expression of p-RhoA, ROCK2 and cleaved-caspase-3 was up-regulated ( P<0.05), and the pathological injury to hippocampal neurons was found in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, the apoptosis rate of hippocampal neurons and [Ca 2+ ] i were decreased, the expression of p-RhoA, ROCK2 and cleaved-caspase-3 was down-regulated ( P<0.05), and the pathological injury to hippocampal neurons was attenuated in group Y. Conclusions:The mechanism by which multiple exposures to sevoflurane induces long-term cognitive impairment is related to activation of RhoA/Rock2 signaling pathway and induction of apoptosis rate of hippocampal neurons in neonatal rats.

Objective:To evaluate the role of reactive oxygen species (ROS)-mediated mitochondrial pathway of apoptosis in long-term cognitive impairment induced by multiple exposures to sevoflurane in the neonatal rats.Methods:Sixty SPF healthy neonatal Sprague-Dawley rats, weighing 12-20 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), multiple exposures to sevoflurane for anesthesia group (group S) and ROS inhibitor group (group A). Group S and group A inhaled 3% sevoflurane for 2 h starting from 6, 7 and 8 days after birth, while group C inhaled air.In group A, ROS inhibitor N-acetylcysteine (NAC) 150 mg/kg was intraperitoneally injected before each anesthesia with sevoflurane.The spontaneous activity was evaluated by open field test on day 35 after birth.The cognitive function was determined by Morris water maze test on day 36 after birth.The rats were sacrificed after the end of Morris water maze test, and the hippocampal tissues were obtained for determination of the apoptosis rate of hippocampal neurons, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) (by flow cytometry) and levels of Cyt c and cleaved caspase-9 and caspase-3 (by Western blot). The expression of Bcl-2 and Bax mRNA was detected by real-time polymerase chain reaction.The ultrastructure of mitochondria in hippocampal neurons was observed with a transmission electron microscope. Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the apoptosis rate of hippocampal neurons and levels of ROS and MMP were increased, the expression of Cyt c, cleaved caspase-9, cleaved caspase-3 and Bax mRNA was up-regulated, the expression of Bcl-2 mRNA was down-regulated, the ratio of Bax/Bcl-2 was increased ( P<0.05), mitochondria were swollen, and mitochondrial cristae structure was broken in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, the apoptosis rate of hippocampal neurons and levels of ROS and MMP were decreased, the expression of Cyt c, cleaved caspase-9, cleaved caspase-3 and Bax mRNA was down-regulated, the expression of Bcl-2 mRNA was up-regulated, the ratio of Bax/Bcl-2 was decreased ( P<0.05), and the mitochondrial swelling and rupture of cristae structure were improved in group A. Conclusion:The mechanism by which multiple exposures to sevoflurane induce long-term cognitive impairment may be related to activating the ROS-mediated mitochondrial pathway of apoptosis in neonatal rats.

Objective:Based on the Delphi method, the electronic intelligent kanban system for general ward nursing was constructed, and its clinical application was studied.Methods:This study was the quasi experimental research, 39 nursing staff working in the experimental ward of electronic intelligent kanban in general surgery of Zhongshan Hospital Fudan University in November 2021 and November 2022 were selected, Delphi expert consultation was used to develop the menus and detailed contents of general ward nursing electronic intelligent kanban system, and it was used in clinical practice. We used the Chinese version of the Nursing Assessment of Shift Report to evaluate the effect of nursing staff before and after the application.Results:The authority coefficients of the two rounds of expert consultation were 0.868 and 0.886 respectively, and the Kendall coordination coefficients of expert consultation were 0.068 and 0.076 respectively (all P<0.01). Finally, the electronic intelligent kanban consisted of 4 first-level menus, 8 second-level menus and 13 third-level menus. After the application of electronic intelligent kanban, the score of Nursing Assessment of Shift Report increased from (79.62 ± 7.64) to (83.51 ± 2.47), with a statistically significant difference ( t=-3.03, P<0.05). Conclusions:The constructed nursing electronic intelligent kanban system was scientific and reliable, which will be beneficial to improve the the effect of nurse shift handover.

Objective:To evaluate the effect of esketamine on hippocampal neuronal necroptosis in aged rats with postoperative cognitive dysfunction.Methods:One hundred and twenty SPF-grade healthy male Sprague-Dawley rats, aged 22 months, weighing 550-600 g, were divided into 4 groups ( n=30 each) using a random number table method: control group (group C), postoperative cognitive dysfunction group (group P), postoperative cognitive dysfunction+ esketamine group (group PE), and esketamine group (group CE). Rats received exploratory laparotomy under sevoflurane anesthesia, and esketamine 10 mg/kg and the equal volume of 0.9% sodium chloride were intraperitoneally injected at the end of surgery once a day for 6 consecutive days in group P and group PE, respectively. Rats received no anesthesia and surgery, and esketamine 10 mg/kg and the equal volume of 0.9% sodium chloride were intraperitoneally injected at the end of surgery once a day for 6 consecutive days in group CE and group C, respectively. Morris water maze test was performed at 7th day after surgery. The escape latency, times of crossing the original platform and time spent in the original platform quadrant were recorded. The rats were sacrificed at the end of Morris water maze test, and the hippocampal tissues were collected for determination of the rate of necroptosis and cytosolic Ca 2+ concentrations (by flow cytometry) and expression of mixed lineage kinase domain-like protein (MLKL), phosphorylated MLKL (p-MLKL), receptor-interacting protein kinase-3 (RIPK3), phosphorylated RIPK3 (p-RIPK3), receptor-interacting protein kinase-1 (RIPK1) and phosphorylated RIPK1 (p-RIPK1) (by Western blot). Results:Compared with group C, the escape latency was significantly prolonged, the times of crossing the original platform were decreased, the time spent in the original platform quadrant was shortened, the necroptosis rate of hippocampal neurons and cytosolic Ca 2+ concentrations were increased, and the expression of MLKL, p-MLKL, RIPK3, p-RIPK3, RIPK1 and p-RIPK1 was up-regulated in group P and group PE ( P<0.05). Compared with group P, the escape latency was significantly shortened, the times of crossing the original platform were increased, the time spent in the original platform quadrant was prolonged, the necroptosis rate of hippocampal neurons and cytosolic Ca 2+ concentrations were decreased, and the expression of MLKL, p-MLKL, RIPK3, p-RIPK3, RIPK1 and p-RIPK1 was down-regulated in group PE ( P<0.05). Conclusions:The mechanism by which esketamine attenuates postoperative cognitive dysfunction may be related to inhibition of necroptosis in hippocampal neurons of aged rats.

Objective:To evaluate the effect of transcutaneous auricular vagus nerve stimulation (taVNS) on tourniquet-induced hypertension (TIH) in the patients undergoing anterior cruciate ligament reconstruction.Methods:Seventy-four patients of either sex, aged 18-60 yr, of American Society of Anesthesiologists Physical Status classification I or II, with body mass index of 18-30 kg/m 2, undergoing elective anterior cruciate ligament reconstruction under general anesthesia combined with preoperative femoral nerve block, were divided into 2 groups ( n=37 each) using a random number table method: sham stimulation group (group SS) and group taVNS. Group SS received stimulation on the ear lobe and the tail of the helix of the left ear. Group taVNS received stimulation on the cymba concha and the earlobe of the left ear. Both groups received stimulation from 1 h before induction of anesthesia until the end of the procedure (frequency of 30 Hz, pulse width of 300 μs, and amplitude of the strongest current that could be tolerated by the patient in the absence of pain). The tourniquet inflation pressure was 280 mmHg, with an inflation time of 60-90 min. Systolic blood pressure, diastolic blood pressure and heart rate were recorded before tourniquet inflation to assess the development of intraoperative TIH. The consumption of intraoperative propofol, remifentanil, nitroglycerin, esmolol, norepinephrine and atropine was recorded, and the occurrence of postoperative nausea and vomiting, skin itching and headache and dizziness was also recorded. Results:Compared with group SS, the incidence of TIH and the number of patients used nitroglycerin were significantly reduced ( P<0.05), and no significant changes were found in the other parameters in group taVNS ( P>0.05). Conclusions:taVNS can decrease the occurrence of TIH in the patients undergoing anterior cruciate ligament reconstruction.