Objective To evaluate the role of etomidate post-conditioning on mitochondrial permeability transition pore (mPTP) in the rat cortical neurons subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with Robo receptors.Methods The cortical neurons obtained from Sprague-Dawley rats (＜ 24 h after birth) were cultured in vitro and seeded in 6-well plates (2 ml/well).The neurons were divided into 4 groups (n=24 each) using a random number table:control group (group C),OGD/R group,etomidate post-conditioning group (group E),and etomidate post-conditioning + Robo receptor blocker group (group ER).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In E and ER groups,etomidate was added to the culture medium with the final concentration of 6 μmol/L immediately after onset of O2-glucose supply.In group ER,Robo blocker RoboN was added to the culture medium with the final concentration of 1 μg/ml at 6 h before O2-glucose deprivation.The neuronal apoptosis was detected using Hoechst/PI double staining,the viability of neurons was measured by MTT assay,and the amount of lactic dehydrogenase (LDH) released was measured using colorimetric method.The mitochondria were extracted,and mitochondrial permeability transition pore (mPTP) opening was detected.Results Compared with group C,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in OGD/R,E and ER groups (P＜0.05).Compared with group OGD/R,the apoptosis rate,amount of LDH released,and mPTP opening were significantly decreased,and the cell survival rate was increased in group E,and the apoptosis and amount of LDH released were significantly decreased,and the cell survival rate was increased in group ER (P＜0.05).Compared with group E,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in group ER (P＜0.05).Conclusion Etomidate post-conditioning mitigates OGD/R-induced damage to the cortical neurons through activating Robo receptors and inhibiting mPTP opening in rats.

Objective To study the related factors of no or weak immune response after inoculation of hepatitis B vaccine .Meth‐ods 120casesofadultreceivedinoculationofhepatitisBvaccineinourhospitalwereenrolled,including60casesofnoweakre‐sponse and 60 cases of normal response ,they were divided into non response group and response group .Related factors of no or weakimmuneresponsewereanalyzed.Results (1)singlefactoranalysis:Therewerestatisticalsignificanceofage,gender,BMI, smoking history ,HBV potential infection ,CD4+ T cells and CD8+ T cell content ,IFN‐γand IL‐2 content between two groups ;(2) logistic regression analysis :old age ,male ,smoking history ,high values of BMI ,HPV latent infection ,low content of CD4+ T cells and IFN‐γand IL‐2 ,high content of CD8+ cells were risk factors of no or weak immune response after inoculation of hepatitis B vaccine .Conclusion No or weak immune response after inoculation of hepatitis B vaccine is affected by gender ,age ,smoking ,obesi‐ty ,potential infection of HPV and immune function ,and necessary measure should be taken .

In this study ,we intended to prepare anti‐Toxoplasma gondii aquaporin (TgAQP) peptide antibody which was used to the application in the detection of the aquaporin expression and its subcellular localization of Toxop lasma gondii (T .gondii) ME49 strain .The B cell peptide antigen was designed based on the TgAQP amino acids sequence .After the pep‐tide antigen was conjugated to the KLH ,the fusion antigen was injected into New Zealand rabbits to prepare polyclonal anti‐body ,followed by identification of ELISA ,Western‐blotting and immunofluorescence assays .The ELISA showed that the titer of anti‐TgAQP antibody was about 1∶40 000 .Western blotting revealed the specific affinity of the antigen to polyclonal anti‐body at 29 .9 kDa protein T .gondii .The protein detected by the indirect immunofluorescence assays was distributed in the cy‐toplasm of the parasite .Thus far ,the anti‐TgAQP polyclonal antibody was successfully prepared ,providing a useful tool for further study of biological function and metabolic characteristics of TgAQP .

Objective To evaluate the role of mitochondrial ATP-seusitive potassium (mito-KATP) channels in sevoflurane postconditioning-induced inhibition of oxygen-glucose dcprivation and restoration (OGD/R)-induced pyroptosis in primary rat cardiomyocytes.Methods Cardiomyocytes of newborn Sprague-Dawley rats (＜48 h after birth) were cultured in vitro and seeded in 6-well dishes (2 cm in diameter)or in 96-well plates.The cells were divided into 6 groups (n =15 each) using a random number table:control group (group C),OGD/R group (group O),sevoflurane postconditioning group (group Sev),sevoflurane postconditioning plus 5-hydroxydecanoate (5-HD) group (group SH),5-HD group (group H) and OGD/R plus 5-HD group (group HO).The cardiomyocytes were subjected to oxygen-glucose deprivation for 4 h followed by restoration of oxygen-glucose supply for 24 h.After oxygen-glucose restoration,the cardiomyocytes in the culture media were exposed to 2％ sevoflurane for 1 h to perform sevoflurane postconditioning.At 1 h before oxygen-glucose deprivation,a specific mito-KATP channel blocker 5-HD 100 μmol/L was added to the culture media.Cardiomyocytes were cultured in normal culture atmosphere in group C.Cardiomyocytes were collected at 24 h of oxygen-glucose restoration.Cell pyroptosis was detected by double flow cytometry AlexaFour488 (caspase-1 FLICA staining) and TMR red (DNA staining) staining.The pyroptosis rate was calculated.The cell survival rate was measured by methyl thiazolyl tetrazolium assay.The content of reactive oxygen species (ROS) in mitochondria was determined by 2',7'-dichlorofluorescin diacetate assay.The mitochondrial membrane potential (MMP) was measured by using JC-I fluorescent probe.The expression of interleukin-1beta (IL-1β) was determined by Western blot.Results Compared with group C,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group O (P＜0.05).Compared with group O,the pyroptosis rate and ROS content were significantly decreased,the cell survival rate and MMP were increased,and the expression of IL-1β was down-regulated in group Sev (P＜0.05).Compared with group Sev,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and M MP were decreased,and the expression of IL-1β was up-regulated in group SH (P＜0.05).Compared with group SH,the pyroptosis rate and ROS content were significantly increased,the cell survival rate and MMP were decreased,and the expression of IL-1β was up-regulated in group H O (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning inhibits OGD/R-induced pyroptosis in primary rat cardiomyocytes is probably associated with increasing mito-KATP channel opening.

AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmid for expression in E. coli. METHODS: Total fragment of hSP17 cDNA gene were amplified by RT-PCR, then subcoloned into pGEX-3b to generate recombinant hSP17/pGEX. Right orientation of insert are identified by restricted enzyme digestion. Transform the correct recombinant plasmid into the E. coli DH5a. The expression of fusion proteins hSP17-GST were induced by adding isopropylthiogalactoside (IPTG). RESULTS and CONCLUSION: The recombinant plasmid hSP17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected.

Objective To analyze the risk factors of nosocomial infection in Department of Neurosurgery and to provide evidence for the prevention and treatment of infection.Methods A total of 931 patients with neurosurgery operation in our hospital from January 2012-January 2016 were collected medical history data immediately after admission,including age,gender,underlying diseases,and primary diseases.Surgical records include preoperative white blood cell count,blood glucose level before operation,duration of operation,and reoperation.Hospitalization records include hospitalization time,without the use of corticosteroids,with or without the use of proton there is no pump inhibitor,and tracheal intubation / incision.Patients were divided into infection group and non infection group according to whether the hospital infection occurred during hospitalization.The difference of two groups of clinical data with statistically significant variables was Logistic multivariate regression analysis.Results There were 112 patients with nosocomial infection,the infection rate was 12.03％,and the infection occurred in the postoperative 3-25 d.The main infection site was postoperative wound,accounting for 35.7％;respiratory tract,accounting for 34.8％.There were 64 strains of pathogenic bacteria,81 strains of Gram-negative bacteria,accounting for 64.1％,21 strains of gram positive bacteria,accounting for 32.8％,2 strains of fungi,accounting for 3.1％.There were significant difference between infection group and non infection group in ≥ 60 years,with basic diseases,reoperation,combined with other injuries,white blood cells,abnormal preoperative hyperglycemia,glucocorticoid use,proton pump inhibitors use,tracheotomy,hospitalization time,operation time (P ＜ 0.05).Further Logisitc regression analysis showed that age,reoperation,hospitalization time,preoperative high blood sugar and tracheotomy were the risk factors of nosocomial infection in Department of neurosurgery.Conclusions For the older,reoperation,longer hospitalization time,preoperative hyperglycemia and tracheotomy patients can take specific measures to improve the immunity of the patients,the rational use of antimicrobial drugs to avoid the occurrence of postoperative infection.

Objective To explore the relationship of sevoflurane neurotoxicity with the expres-sion of Bid,Bim,Puma.Methods The cortical neuron from newborn SD rat (within 24 h)were see-ded in 6 or 12 well plate,and then randomly divided into 4 groups.Rat culture cortical neurons in vitro exposed in 1%,2%,4% and 0% sevoflurane for 6h were divided into A,B,C and D group. The effect of neuron viability,death and apoptosis were assessed using CCK-8,LDH and caspase-3 cleavage 1 7kDa expression assay.The expressions of Bid,Bim and Puma were assessed by western blot.Results Compared with group D, there were significant increases of neuron death and apoptosis,but a decrease of neuron viability,and upregulated expressions of Bid,Bim and Puma in group B (P <0.05);Compared with group B,Group C had increased death and apoptosis and de-creased viability of neurons,as well as upregulated expressions of Bid,Bim and Puma (P <0.05 ). Conclusion Along with the increase of the concentration,sevoflurane neurotoxicity was increased by upregulation of Bid,Bim,Puma expression.

Objective To observe the sedation and analgesia in surface anesthesia in conscious glossopha-ryngeal LMA for the clinical effect of laparoscopic hysterectomy.Methods 90 patients undergoing elective laparo-scopic total hysterectomy were randomly divided into three groups,30 cases in each group.The observation group (group A):sedation,analgesia,full surface anesthesia,control group 1 (B group)control group 2 (group C)were treated with endotracheal intubation under general anesthesia.At intubation and pull tube stage,the patients'reaction, hemodynamic changes,pneumoperitoneum 1 h blood gas and perioperative complications were observed.Results The mean arterial pressure(MAP)and heart rate(HR)of A group were (92.7 ±10.6)mmHg and (82.8 ±12.1)/min. Those of B group were (98.4 ±11.6)mmHg,(89.1 ±11.4)/min,C group were (111.2 ±12.5)mmHg,(104.1 ± 13.2)/min,those in A group and B group were better than C group(A group and C group,t =6.18,6.52,P <0.01;B group and C group,t =4.11,4.71,all P <0.01).The pulse oxygen saturation(SpO2 )and peak airway pressure (Paw)of A group were (99.6 ±0.4)and (17.3 ±2.1)mmHg,those in group B were (99.5 ±0.5)% and (17.6 ± 2.0)mmHg,group C were (99.5 ±0.5)% and (22.5 ±2.8)mmHg.The differences between A group and B group were statistically significant(compared with C group,t =14.3,8.14,all P <0.01;B group and C group,t =12.7, 7.78,all P <0.01).The incidence rates of perioperative complications and adverse reactions of A,B,C groups were 27%,33%,67%,that in C group was significantly higher than A group and B group(χ2 =9.64,6.67,all P <0.01). Conclusion The laryngeal mask airway was used for laparoscopic total hysterectomy under the condition of sedation and analgesia,and it can shorten the time of the whole body,prevent the difficulty of intubation,emergency airway and anesthesia related complications.

Objective To evaluate the role of extracellular signal-regulated kinase (ERK) signaling pathway in inhibition of oxygen-glucose deprivation and restoration (OGD/R)-induced apoptosis in rat cortical neurons by sevoflurane and the relationship with mitochondrial permeability transition pore (mPTP).Methods The rat cortical neurons were cultured in vitro and seeded in 6-well or 12-well culture plates.The neurons were randomly divided into 4 groups (n =18 each) using a random number table:control group (group C);OGD/R group (group O);sevoflurane group (group OS);sevoflurane + ERK1/2 inhibitor PD98059 group (group OSP).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h in group O.The neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OS.OGD/R was performed at 1 h after ERK1/2 inhibitor PD98059 30 μmol/L was added,and the neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OSP.At 24 h of restoration of O2-glucose supply,the expression of phosphorylated ERK1/2 (p-ERK1/2) in neurons was measured by Western blot,the neuronal apoptosis was detected using Annexin V-FITC/PI double staining combined with flow cytometry,and the opening of mPTP was determined through measuring the optical density at 540 nm.The apoptosis rate was calculated.Results Compared with group C,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly increased in group O (P＜0.05).Compared with group O,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly decreased in group OS (P＜0.05).Compared with group OS,the expression of p-ERK1/2 in neurons was significantly down-regulated,and the apoptosis rate and mPTP opening were significantly increased in group OSP (P＜0.05).Conclusion The mechanism by which sevoflurane inhibits OGD/R-induced apoptosis in rat cortical neurons is related to inhibition of mPTP opening after activation of ERK signaling pathway.