ObjectiveTo investigate the effects of sevoflurane on short-term memory impairment and the relative synaptic mechanism.MethodsAt 12 h,24 h and 72 h after the mice were exposed to 1.5％ sevoflurane for 1h,the spontaneous alternation and locomotor activity was assessed by Y maze,the short term potentiation (STP) were measured with extracellular recording technique in hippocampal slices.ResultsAt 12h after administration with sevoflurane in vivo,the spontaneous alternation ( (54.7 ± 1.7) ％,P＜0.01 ) and locomotor activity (16.4 ± 1.3,P ＜ 0.01 ) decreased significantly compared with that of control group,and the population spike amplitude after induction of STP ( ( 122.3 ± 13.9) ％,P ＜ 0.05 ) decreased significantly in hippocampal slices,but there was no different at 24h and 72h.After administration with sevoflurane in vitro,the basic ( ( 83.5 ± 8.9) ％,P ＜ 0.01 ) or titanic ( ( 116.5 ± 14.9) ％,P ＜ 0.01 ) population spike amplitude decreased significantly in hippocampal slices,but the amplitude could recovery after wash-out.ConclusionSevoflurane can impair the shortterm memory by suppressing synaptic transmission in the near future but not the long future.

BACKGROUND: Lidocaine is deemed to can attenuate the inflammatory response, but the mechanisms are poorly understood. Nuclear factor-κB (NF-κB) is a key process of imflammatory response. OBJECTIVE: To determine the effect of lidocaine on activation and apoptosis of NF-κB in human neutrophiis in vitro. DESIGN, TIME AND SETTING: The controlled experiment was conducted at the Jiangsu Provincial Key Laboratory of Anesthesiology from October 2006 to February 2007. MATERIALS: Tumor necrosis factor (TNF)-α (0127:B8) was purchased from Sigma, USA. Peripheral blood samples were obtained from healthy donors after their informed consent. METHODS: Human neutrephilic granulocytes were assigned into five groups: ① saline control, ② TNF-α, ③ TNF-α+ lidocaine 1.0 mmol/L, ④ TNF-α+ lidocaine 2.0 mmol/L, and ⑤ TNF-α+ lidocaine 4.0 mmol/L groups. Incubation wasperformed for 3 hours.MAIN OUTCOME MEASURES: The effects of lidocaine on expression of NF- κB mRNA and I- κB mRNA in the cytosol extracts were analyzed by reverse transcription-polymerase chain reaction (RT-PCR), and content of p65 protein were analyzed by Western blot. Neutrophilic granulocyte apoptosis was detected on flow cytomatry after incubation 12 hours and 24 hours.RESULTS: The expression of NF- κB mRNA in the nuclear extracts was significantly decreased and I- κB mRNA in the cytosoI extracts was markedly increased in the lidocaine group. The expression of NF- κB was significantly better in the 2.0 mmol/L and 4.0 mmol/L lidocaine groups than in the 1.0 mmol/L lidocaine group (P< 0.05). No significant difference was detected between the 2.0 mmol/L lidocaine group and the 4.0 mmol/L lidocaine group (P0.05). Lidocaine could significantly inhibit peripheral blood neutrophilic granulocyte apoptosis induced by TNF-α (p < 0.05). The inhibitory effect was significantly better in the 4.0 mmol/L lidocaine group than in the 1.0 mmol/L lidocaine group (P < 0.05). CONCLUSION: Lidocaine (1.0, 2.0 and 4.0 mmol/L) can down-regulate the expressions of NF- κB subunit p65 mRNA and the content of p65 protein in human polymorphoneclear neutrophils, and can significant reverse the reduction of apoptosis induced by TNF-α.

Objective To establish a doxycycline-controlled immortalized pre-cartilaginons stem cells (IPCSCs) strains, clone parathroid hormone-related peptide[PTHrP(1-36)] gene and construct re- sponsive plasmid, pTRE-PTHrP (1-36). Methods Plasmid pTet-on was transfected into IPCSCs by using LipoinfectaminTM 2000 and then the stable clones were obtained by G418 screening. The doxycyc- line was added into the medium of monoclonal cells that were transiently transfected with plasmid pTRE- 2Hyg-Lue. The total RNA was extracted from PCSCs and the PTHrP(1-36) gene obtained by RT-PCR method. Then, the PTHrP (1-36) gene was subcloned to plasmids of Tet-responsive element with the se- lection marker of hygromycin pTRE-2Hyg to construct recombinant eukaryotic expression plasmid pTRE- PTHrP(1-36). After transferred into E. coli-DH5α, the clone was amplified, the recombinant plasm0ids were purified and identified by double-enzyme digestion. Results The doxycycline induced IPCSCs line was obtained, with 50 times higher than the non-induced cell line. Double enzyme digestion analysis and sequencing showed that the target gene was cloned into recombinant plasmid. Conclusions The induced IPCSCs line can be used to highly express alien genes. The responsive plasmid containing PTHrP (1-36) gene may be premising for rigorous control of PTHrp (1-36) gene expression.

Objective To investigate the effect of sevoflurane anesthesia on learning and memory in mice and the relationship with histone acetylation. Methods Thirty-six adult male C57BL/6 mice were ran-domly divided into control group(mice inhaled 95％ O2 for 6 h), sevoflurane (Sevo) group (1. 5％ Sevo group, 2％ Sevo group, 3％ Sevo group:mice inhaled 1. 5％, 2％ and 3％ sevoflurane for 6 h respectively) , sevoflurane + SAHA (Sevo + SAHA) group (mice were intraperitoneally injected with histone deacetylase inhibitor SAHA ( 25 mg/kg) . And 1 h later, 3％ Sevo was inhaled continuously for 6 hours. ) and SAHA group(mice were intraperitoneally injected with histone deacetylase inhibitor SAHA (25 mg/kg)). The abil-ity of learning and memory in mice was estimated by Morris water maze. The expression levels of Ac-H3, BDNF and Syt-I protein in the hippocampus were detected by Western blot. Results In Morris water maze, 3％ sevoflurane anesthesia significantly prolonged the escape latency((46. 91±1. 84)s),and significantly decreased the ratio of target time((35. 84±5. 40)％) compared with that of control group((23. 46±2. 67)s, (49. 74±4. 91)％,P<0. 05). Compared with 3％ Sevo group,the ratio of target time in Sevot+SAHA group ((46. 86±4. 37)％) was increased(P<0. 05). Moreover,3％ sevoflurane anesthesia significantly decreased the expression levels of Ac-H3 (10. 23±2. 45),BDNF (6. 72±1. 21) and Syt-I (8. 25±2. 11) in the hippo-campus compared with that of control group((15. 45±2. 58),(10. 17±1. 45) and (15. 02±3. 38),P<0. 05) . However,pre-administration of the histone deacetylase inhibitor SAHA significantly increased the ra-tio of target time in Morris water maze,and improved the expression levels of Ac-H3 (14. 06±2. 79),BDNF (10. 13±1. 06) and Syt-I (14. 16±3. 66) in Sevo+SAHA group compared with that of Sevo group (P<0. 05) . Conclusion The high dose of sevoflurane anesthesia can induce learning and memory impairment through the inhibitation of histone acetylation in the hippocampus.

Objective:To explore the improvement and its mechanism of minocycline on sevoflurane anesthesia induced cognitive dysfunction in aged mice.Methods:Totally 75 aged clean-grade C57BL/6 mice were randomly divided into control (Con) group( n=25), sevoflurane (Sev) group( n=25) and sevoflurane + minocycline (Sev+ Min) group( n=25). Anesthetic injury was induced by 3% sevoflurane (2 h/d for 3 days) in Sev group. Minocycline (50 mg/kg) was injected intraperitoneally first, and then 3% sevoflurane (2 h/d for 3 days) anesthesia was performed in Sev+ Min group. Saline alone was injected intraperitoneally (once a day for 3 days) in Con group.The spatial memory function was detected by Morris water maze experiments. BrdU was used to label new neuron and the proliferation was observed by immunohistochemistry. The field excitatory postsynaptic potential (fEPSP) slope was measured in hippocampal dentate gyrus (DG) region of isolated brain slices by electricphysiological technique.The data were analyzed by repeated measures analysis of variance and SNK-q test using SPSS 21.0 software. Results:Results from positioning navigation experiment showed that the group×time interaction effect of mice was significant( F=15.65, P<0.01). On the 6th day after anesthesia, compared with Con group, the escape latency of the original platform in Sev group was significantly increased ( q=4.35, P<0.05) in space exploration experiment, while the target quadrant time ratio ( q=6.15, P<0.05))and the mean annulus crossings ( q=6.45, P<0.05) were significantly decreased. Compared with Sev group, the escape latency in Sev+ Min group was significantly decreased ( q=3.01, P<0.05), while the target quadrant time ratio ( q=3.21, P<0.05) and the mean annulus crossings ( q=3.48, P<0.05) were significantly increased. In immunohistochemistry experiment, the number of BrdU positive cells in Sev group was significantly reduced ((227.45±43.25), q=8.67, P<0.01) compared with Con group (355.87±62.58). Compared with Sev group, the number of BrdU positive cells in Sev+ Min group was significantly increased ((338.73±47.27), q=8.68, P<0.01). In electricphysiological test, the fEPSP slope after high frequency stimulation in Sev group ((126.83±25.67)%, q=6.18, P<0.01)) was significantly lower than that in Con group((214.38±43.42)%). In Sev+ Min group, the fEPSP slope was significantly higher ((178.49±32.67)%, q=3.64, P<0.05) than that in Sev group. Conclusion:Sevoflurane anesthesia can induce the short-term cognitive dysfunction in aged mice, and its mechanism is related to inhibiting neuron proliferation and synaptic plasticity. Minocycline can alleviate the damage caused by sevoflurane.

Objective To study the effects of pressure controlled ventilation(PCV)and volume controlled ventilation(VCV)on diaphragm function and the incidence of postoperative pulmonary complica-tions(PPCs)in patients undergoing laparoscopic surgery.Methods Sixty-six patients underwent laparo-scopic gynecological surgery under general anesthesia,aged 18-64 years,BMI 18-30 kg/m2,ASA physical status Ⅰ or Ⅱ were recruited.The patients were randomly divided into two groups:PCV group(group P)and VCV group(group V),33 cases in each group.All the patients were ventilated in VCV mode after induction.Group P was switched to PCV after pneumoperitoneum and group V maintained VCV until the end of operation after pneumoperitoneum.The diaphragm ultrasonic evaluation indexes including di-aphragmatic excursion(DE),diaphragm contraction velocity(DCV),and diaphragmatic rapid shallow breathing index(D-RSBI)were recorded before anesthesia induction,immediately after extubation,and 30 minutes after extubation.The mechanical ventilation time,artificial pneumoperitoneum time,the time from the end of artificial pneumoperitoneum to extubation,the cumulative dosage of cisatracuriumbesylate,and the patient's observer's assessment alert/sedation(OAA/S)immediately after extubation,the incidence of diaphragm dysfunction immediately after extubation and 30 minutes after extubation,and the cumulative in-cidence of PPCs in 1-3 days after operation.Results Compared with group V,DE in group P was in-creased significantly immediately after extubation(P＜0.05),but there was no significant difference in DE of 30 minutes between the two groups after extubation.Compared with group V,DCV in group P was in-creased significantly immediately after extubation and 30 minutes after extubation(P＜0.05),the inci-dence of PPCs in group P was significantly lower on the 1st day after operation(P＜0.05).There were no significant differences in D-RSBI,time of mechanical ventilation,time of artificial pneumoperitoneum,time from the end of pneumoperitoneum to extubation,cumulative dosage of atracurium besylate,OAA/S score immediately after extubation,and the incidence of diaphragm dysfunction immediately after extubation and 30 minutes after extubation,and the cumulative incidence of PPCs on the 2nd and 3rd day after operation.Conclusion In lower abdominal endoscopic gynecological surgery,compared with volume-controlled venti-lation mode,pressure-controlled ventilation mode dose not reduce the incidence of postoperative diaphragm dysfunction,but dose alleviate the weakening of diaphragm inspiratory force and reduce the incidence of pul-monary complications on the first day after operation.

Objective:To investigate changes of intestinal flora and the mechanism of NLRP3 inflammasome in elderly mice with cognitive dysfunction induced by sevoflurane anesthesia.Methods:Eighteen fourteen-month-old male SPF grade C57BL/6J mice were randomly divided into control and sevoflurane groups, with 9 mice in each group. The mice of sevoflurane group inhaled 3% sevoflurane for 2 hours daily for three days. Fecal samples were collected post-exposure 24 hours for 16S rRNA sequencing. Morris water maze was then used to test the cognitive ability. Western blot was used to detect the expressions of synapse-associated proteins, NLRP3 inflammasome-related proteins of hippocampus, and NLRP3 inflammasome-related proteins of colon. Golgi staining was used to observe the number of dendritic spines in the hippocampus. qPCR was used to detect the expression of inflammatory cytokines IL-1β, IL-18, TNF-α mRNA in mice colon and hippocampal tissues.Results:(1) The Morris water maze test showed that the escape latency of the sevoflurane group was longer than the control group, but there was statistical difference only on the fifth day ( P<0.05). In the spatial exploration test, escape latency of the sevoflurane group was higher than that of the control group((49.50±9.99)s, (18.67±7.63)s, t=6.005, P<0.001), and platform crossing frequency was less than that of the control group((0.83±0.75)times, (2.33±1.03)times, t=2.87, P=0.017). (2) Western blot and Golgi staining results showed that the expression of hippocampal synaptic-related proteins and the number of dendritic spines in the sevoflurane group were significantly reduced compared with those in control group (all P<0.05). (3) 16S rRNA sequencing showed significant β-diversity difference between the two groups ( P<0.05). Compared with the control group, potential pathogens that p_Desulfobacterota and g_Desulfovibrio increased significantly in the sevoflurane group (both P<0.05), and beneficial bacteria that p_Verrucomicrobiota and g_Akkermansia decreased significantly (both P<0.05). (4) Compared with the control group, the results of qPCR showed increased expression of inflammatory cytokines TNF-α, IL-1β mRNA in the colon and hippocampal tissues of the sevoflurane group (all P<0.05). Western blot results showed increased expression of NLRP3 inflammasome-related proteins in the colon and hippocampal tissues of the sevoflurane group (both P<0.05). Immunofluorescence results showed the higher fluorescence intensity of ASC in the DG region of the hippocampus of the sevoflurane group compared with the control group ( P<0.01). Conclusion:The cognitive dysfunction model induced by sevoflurane in elderly mice shows neuroinflammatory reactions and synaptic damage, which may be related to intestinal microbiota imbalance and activation of NLRP3 inflammasome.

Objective:To evaluate the effect of dexamethasone combined with aminophylline on perioperative airway responses in COVID-19 convalescent patients undergoing gynecological laparoscopic surgery.Methods:Sixty-eight COVID-19 convalescent patients, aged 25-57 yr, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, undergoing gynecological laparoscopic surgery, were divided into experimental group ( n=34) and control group ( n=34). In experimental group, dexamethasone 10 mg was intravenously injected at the beginning of anesthesia induction, and aminophylline 0.25 g (in 100 ml of normal saline) was intravenously infused for 10 min starting from 15 min before the end of surgery. In control group, the equal volume of normal saline was administrated instead at the same time point. Airway secretions, laryngospasm and bronchospasm were recorded from the time point before operation to 24 h after operation, and coughing was also recorded from emergence to 3 min after extubation. The blood eosinophils (EOS) count, percentage of EOS (EOS%), and neutrophil to lymphocyte ratio were determined, and plasma C reactive protein level was measured by enzyme-linked immunosorbent assay before operation and at 24 h after operation. The serum levels of interleukin-6, tumor necrosis factor-α and interferon-γ were measured before operation, at 5 and 10 min after extubation and at 24 h after operation. Results:Compared with control group, the incidence of coughing, severity of coughing, incidence of increased airway secretion, and grade of airway secretion were significantly decreased, the levels of EOS, EOS%, neutrophil to lymphocyte ratio and plasma C reactive protein in peripheral blood and serum levels of interferon-γ, interleukin-6 and tumor necrosis factor-α were significantly decreased after operation ( P<0.05), and no significant change was found in the incidence of bronchial spasm in experimental group ( P>0.05). No laryngeal spasm occurred in both groups. Conclusions:Dexamethasone combined with aminophylline can relieve the perioperative airway responses by inhibition of inflammatory responses in COVID-19 convalescent patients undergoing gynecological laparoscopic surgery.