Objective:Biological markers of the ventrolateral orbitofrontal cortex(vlOFC)involved in pain regula-tion were screened.Methods:Chronic inflammatory pain was induced in male C57BL/6J mice by injection of complete Freund's adjuvant(CFA)into the left posterior plantar.Paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)were detected to evalue hyperalgesia.Transcriptome sequencing was performed on fresh tissue from vlOFC of mice after behavioral tests.The differentially expressed genes(DEGs)were screened by bioinformatics method,and their biological functions and pathways were enriched.Results:Compared with the PBS group,the left hindpaw me-chanical pain threshold and the paw withdrawal latency caused by heat pain were significantly reduced in the CFA group(P＜0.001).The DEGs of vlOFC in the two groups were 497,of which 143 were up-regulated and 354 were down-reg-ulated.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGGs)analysis showed that:In chro-nic inflammatory pain model mice,DEGs of vlOFC were mainly manifested in biological processes such as organic cation transport,neurotransmitter transport,and regulation of cytoplasmic calcium ion concentration.It is related to G protein-coupled receptors(GPCRs),neuropeptides and ammonium transport.DEGs mainly focuses on neuroactive ligand-receptor interactions,cytokine-cytokine receptor interactions,and cAMP signaling pathways.Reactome functional en-richment analysis showed that the pathway with the highest number of DEGs enriched and the lowest P value-adjusted was GPCRs ligand binding.Conclusion:Ion transport,neurotransmitter transport and binding,and GPCRs-related ac-tivities in vlOFC are involved in the regulation of chronic inflammatory pain.

Objective:To evaluate the efficacy of ultrasound-guided adductor canal approach to nerve block using one-puncture four-point technique for analgesia after total knee arthroplasty (TKA).Methods:Sixty patients of both sexes, aged 65-80 yr, with body mass index of 18-27 kg/m 2, of American Society of Anesthesiologists physical status Ⅱor Ⅲ, scheduled for elective unilateral TKA under general anesthesia, were enrolled in this study.Peripheral nerve block was performed for postoperative analgesia under ultrasound guidance.All the patients were allocated into 2 groups ( n=30 each) using a random number table method: adductor canal block combined with posterior approach to sciatic nerve block group (group ASB) and adductor canal approach to nerve block using one-puncture four-point technique including sciatic nerve, medial vastus muscle nerve, saphenous nerve, anterior femoral cutaneous nerve at adductor canal level group (group ANSB). When visual analogue scale score ≥4 during postoperative movement, ketorolac tromethamine 0.6 mg/kg was intravenously injected as rescue analgesic. The muscle strength of quadriceps on the affected side, range of knee joint motion and rate of the ability to walk for 10 m using the walker were recorded at 2, 8, 24 and 48 h after surgery.The consumption of analgesics for rescue analgesia, manipulation completion time, length of hospital stay and patients′ satisfaction score were recorded within 48 h after surgery. Results:Compared with group ASB, the consumption of ketorolac tromethamine was significantly reduced, manipulation completion time and length of hospital stay were shortened, patients′ satisfaction scores were increased, and the range of knee joint motion and rate of the ability to walk for 10 m using the walker were increased at 2, 8, 24 and 48 h after surgery in group ANSB ( P<0.05). There was no significant difference in muscle strength of quadriceps between two groups ( P>0.05). Conclusion:Ultrasound-guided adductor canal approach to nerve block using one-puncture four-point technique provides better efficacy than adductor canal block combined with posterior approach to sciatic nerve block when used for analgesia after TKA.

Objective To compare the effectiveness and feasibility of oral tracheal intubation with Clarus video stylet,HPHJ-A video laryngoscope and Airtraq laryngoscope and in snoring pa-tients.Methods Ninety patients undergoing elective snoring surgery with general anesthesia,3 9 males and 51 females,aged 22-55 years,BMI 25-29 kg/m2,ASA physical status Ⅰ-Ⅲ,were ran-domly divided into three equal-sized groups (n=30):Clarus Video Stylet group (group C),HPHJ-A video laryngoscope group (group H)and Airtraq laryngoscope group (group A).The time of success-ful endotracheal intubation and the success rate of initial intubation of all groups were observed.The mean arterial pressure (MAP),heart rate(HR)and Rate-pressure product(RPP)were also monitored before induction of anesthesia(T0),before tracheal intubation(T1),at 0 min(T2),1 min (T3),and 3 min (T4)after intubation,as well as the throat injury and hemorrhage were noted. Results Patients in group C were successful intubated.Two patients in group H and one patient in group A with failed intubation were successfully intubated by using Clarus video stylet.The time re-quired for successful intubation in group C was longer than groups H and A (P<0.05).Compared with T0,the increasing of hemodynamic parameters (MAP,HR,RPP)after induction of all groups were significant(P<0.05).Compared with T1,the HR,MAP and RPP were increased significantly at T2in all groups (P<0.05).Compared with T1,the MAP and RPP were increased significantly at T3in all groups,and the HR were increased significantly at T3in groups H and A (P<0.05).There was no significant difference in the HR between T1and T3in group C.The increase in the HR and RPP at T2 was lower in group C than that in both the groups H and A (P <0.05).The changes of hemodynamic parameters were no significant among the three groups at other time.The differences of sore throat score and hemorrhage were no significant.Conclusion Compared with HPHJ-A video laryngoscope and Airtraq laryngoscope,Clarus video stylet for guiding oral tracheal intubation in snoring patients have less influence on hemodynamic parameters,and have no limited mouth opening. But Clarus Video Stylet spends longer intubating times,and has no obvious advantages on preventing throat injury.

Objective To compare of the application of tracheal intubation guided by Lightwand,Shikani optical stylet,or Macintosh. Methods Ninety ASA Ⅰ~Ⅱ patients undergoing elective general anesthesia were randomly divided into three groups(n = 30):Macintosh group(group M),Lightwand group(group L)and Shi-kani optical stylet group(group S). MAP and HR were recorded at the time of entering the operating room(T1), before intubation(T2),at the immediate time of intubation(T3),3 mins(T4)and 5 mins after intubation(T5). The intubation duration,the first intubation success rate,the number of intubation,and the incidence of complications including sore throat were observed. Results As compared with T1,MAP and HR decreased significantly at T2in the three groups(P<0.05). As compared with T2,HR increased in group L and MAP and HR increased in group S at T3(P<0.05). As compared with group M,MAP and HR at T4and T5were lower in groups L and S(P<0.05). The first intubation time was obviously shorter in group L than in other groups(P<0.05). The incidence of compli-cations was lower in group S(P < 0.05). The first success rate of intubation,the intubation times and the inci-dence of airway complications did not differ significantly among the three groups.(P > 0.05). Conclusions As compared with Macintosh,Lightwand and Shikani optical stylet have less influence on hemodynamic parameters. Lightwand needs shorter intubation time,Shikani optical stylet has the lowest rate of sore throat.

Objective To evaluate analgesic effect of dexmedetomidine (DEX) combined with ropivacaine in single sciatic nerve block after total knee arthroplasty (TKA) with continuous femoral never block.Meth ods One hundred and twenty patients,ASA physical status Ⅰ ～ Ⅱ,who underwent unilateral TKA were randomly allocated into two groups:group D (DEX with ropivacaine);group R (ropivacaine only).Both groups experienced spinal-epidural anesthesia.Continuous femoral nerve block was performed after operation.Rest and movement visual analogue score (VAS),Ramsay sedation scores were recorded.In addition to the date of the analgesia drug dose,the pressed times of PCA,the mobility of knee joint and the force of quadriceps femoris were obtained at different points of time.Results Rest Pain scores (RVAS) of the group D were lower at time point 8,12,24,36,48 h and Ramsay sedation scores were higher at time point 4,8,12 h than that in the group R (P ＜ 0.05).The analgesia drug dose and the pressed times of PCA reduced in the group D.The range of motion (ROM) was better in the group D at 1 d,2 d after operation (P ＜ 0.05).There was no significant difference in the force of quadriceps femoris in both groups.Conclusion Dexmedetomidine combined with ropivacaine in single sciatic nerve block after TKA provided better analgesia when continuous femoral nerve block was performed.Meanwhile,analgesia lasted longer and did not increase the motor nerve blockade.

Objective To compare the application of Shikani optical stylet(SOS)and Clarus Video Sty-let(Tracway)in patient with cervical spine immobilization in tracheal intubation. Method Sixty patients,ASAⅠ~Ⅱ,undergoing cervical internal fixation operation,were randomly divided into Shikani optical stylet group (Group S,n = 30)and Clarus Video Stylet group(Group T,n = 30). MAP,HR and RPP(The rate-pressure product)were recorded at the point before induction of anesthesia(T1),before intubation(T2),at the immediate time of intubation(T3)and 1 min(T4),3 mins(T5),and 5 mins after intubation(T6).The intubation time,one-time success rate of intubation,the number of intubation times and the incidence of sore throat and other complica-tions were observed. Results The one-time intubation time in group S was obviously shorter than that in group T (P<0.05).The incidence of mild sore throat and intubation throat injury rate were lower in group S than those in group T(P<0.05).Compared with those at T1,MAP and RPP decreased significantly at point of T2~T6in both of two groups(P<0.05).There was no significantly difference in MAP,HR and RPP at any points of time between the two groups. The one-time success rate of intubation,the number of intubation times and the incidence of air-way complications in two groups(P>0.05)were no significantly different. Conclusions Compared with Clarus Video Stylet(Tracway),Shikani optical stylet can shorten the intubation time in patients with cervical spine immo-bilization,but no difference was found in regard to the hemodynamic influence on intubation,success rate of intu-bation,the intubation times,the sore throat and the other related complications.

Objective To investigate the effects of sevoflurane ( Sevo ) on dendritic development and the expression of collapsin response mediator proteins ( CRMP ) in the hippocampus of developing rats. Methods Twenty-four neonatal Sprague Dawley (SD) rats at postnatal day 7 (P7) were randomly divided into control group or sevoflurane group ( 12 rat pups for each group) .Rats in the control group were exposed to air for 4 h,whereas rats in the sevoflurane group were exposed to 2.8%sevoflurane for 4 h.The hippocam-pus of some rats were collected,and the expressions of CRMP1,CRMP2 and CRMP4 proteins and phospho-rylation of CRMP2 protein at Ser522,Thr514 and Thr555 were detected by Western blot 6h after exposure ( n=6) .The rest rats were housed till P30,the expression of CRMP1,CRMP2 and CRMP4 proteins in the hip-pocampus were detected by Western blot ( n=6) and the morphology changes of dendrites in the dentate gy-rus ( DG) of hippocampal neurons were detected by Golgi-Cox Staining ( n=6) .Results The expression of CRMP1,CRMP2 and CRMP4 proteins of rats at P7 in the sevoflurane group was decreased by 35.0%( P=0.004) ,27.5%( P=0.015) and 12.0%( P=0.003) ,respectively,and the phosphorylation of CRMP2 pro-tein at Ser522 and Thr514 in the sevoflurane group were increased by 68.3%( P<0.01) ,74.5%( P<0.01) , respectively,6 h after exposure compared with control rats.However,the phosphorylation of CRMP2 protein at Thr555 was not significantly changed after sevoflurane exposure.At P30,both total dendrite length ( P=0.001) and the dendrites length at level 2 and 3 ( P=0.033, P<0.01,respectively) were shorter and the dendritic branching at 120,140 and 160 μm rings in Sholl analysis were less ( P=0.009, P=0.028, P=0.048,respectively) for rats in the sevoflurane group,compared with control rats.There were no significant changes at the expressions of CRMP1,CRMP2 and CRMP4 proteins.Conclusion Sevoflurane inhibits the development of dendrites in the hippocampal DG area of developing rats,which may be related to inhibition of CRMP1,CRMP2 and CRMP4 proteins expression and hyperphosphorylation of CRMP2 Ser522 and Thr514.

[ABSTRACT]AIM:Toinvestigatetheeffectofsevoflurane(Sevo)onthedendriticdevelopmentinprefrontal cortex ( PFC) of neonatal rats and the role of cyclin dependent kinase 5 ( Cdk5 )-collapsin response mediator protein ( CRMP) pathway in it.METHODS:Eighty-eight postnatal day 7 Sprague Dawley ( SD) rats were randomly divided into 4 groups (n=22):Air+NS group, Air+roscovitine (Ros) group, Sevo+NS group and Sevo+Ros group.The rats in Air+NS group and Air+Ros group were exposed to the air for 4 h, while the rats in the other 2 groups were exposed to 2.8%sevoflurane for 4 h.The rats received intraperitoneal injection of 150μL normal saline 15 min before exposure in the Air+NS group and Sevo+NS group, while the rats in the Air+Ros group and Sevo+Ros group received intraperitoneal injection of 150μL roscovitine ( in DMSO solution, 10 mg/kg) 15 min before exposure.At the end of exposure, the corti-ces of the rat brain were collected and the protein levels of P35, P25, Cdk5, CRMP1, CRMP2, CRMP4 and p-CRMP2 Ser522 in PFC were detected by Western blot.On the postnatal day 30, the rat brains were sectioned for Golgi-Cox staining and morphological analysis of dendrites in the PFC neurons.Open-field test and contextual fear conditioning test were per-formed on postnatal days 25~27 and 31~32, respectively.RESULTS:Compared with Air+NS group, the expression of P35 in the Sevo+NS group was significantly decreased, and the expression of P25 was dramatically increased (P<0.05), whereas roscovitine partly reversed the changes above induced by sevoflurane (P<0.05).The expression of Cdk5 was not significantly different among all groups.Compared with the Air+NS group, the expression of CRMP1, 2, and 4 in the Se-vo+NS group were decreased, and the protein level of p-CRMP2 Ser522/CRMP2 was increased ( P<0.05 ) , whereas roscovitine partly reversed the changes above induced by sevoflurane (P<0.05), except for the expression of CRMP2. Compared with Air+NS group, the total dendrite length, secondary dendritic length and interactions on 60 and 80 μm shells in the Sevo+NS group were decreased (P<0.01), whereas roscovitine partly reversed the changes above induced by sevoflurane (P<0.05).Compared with Air+NS group, the percentage of freezing time in the Sevo+NS group was de-creased (P<0.01), whereas roscovitine partly reversed the changes induced by sevoflurane (P<0.05).No significant difference among groups in the open-field test was observed.CONCLUSION:Sevoflurane exposure disturbed dendritic de-velopment of neurons in PFC, learning and memory ability of neonatal rats, which may be mediated by Cdk5-CRMP path-way.

Aim To investigate the effect of isoflurane on the phosphorylation of p38 mitogen-activated protein kinase (MAPK)in the hippocampus of neonatal rats, and the effect of p38 MAPK pathway on isoflurane-in-duced neuronal apoptosis.Methods Forty-eight neo-natal rats on postnatal day 7 were assigned randomly into four groups:DMSO group (group Air +DMSO), p38 MAPK inhibitor SB203580 group (group Air +SB20 ),isoflurane +DMSO group (group Iso +DM-SO),and isoflurane +SB203580 group (group Iso +SB20 ).Rats were exposed to air or isoflurane (volume fraction of 0.01 1 )for 4h.The p38 inhibitor SB203580 (20 nmol)or DMSO (volume fraction of 0.1 )5μl was intraventricularly administered 30 min before the expo-sure.The brains of some rats in each group were per-fused and embedded by paraffin 6h after the exposure. Neuronal apoptosis in the hippocampal CA1 area was detected by terminal deoxyribonucleotide transferase-mediated dUTP nick end labeling (TUNEL)(n =6). The hippocampal tissues of the other rats in each group were dissected 6h after the exposure,and the protein expressions of phospho-p38 (p-p38 ),p38,cleaved caspase-3,phospho-NF-κB (p-NF-κB ),Bcl-2 and Bax were detected by Westem blot (n =6).Results The number of TUNEL positive cells in the hippocam-pal CA1 region in group Iso +DMSO increased by 4.8 fold compared with that in group Air +DMSO (P <0.01 ),while the number of TUNEL positive cells in group Iso +SB20 decreased by 3 /5 compared with that in group Iso +DMSO (P <0.01 ).The protein expres-sion of cleaved caspase-3 in group Iso +DMSO signifi-cantly increasd (P =0.003)compared to that in group Air +DMSO,which was significantly decreasd in group Iso +SB20 (P =0.007 ).In addition,isoflurane also increased the protein expression of p-p38,p-NF-κB and Bax,decreased the level of Bcl-2,and reduced the ratio of Bcl-2 /Bax compared with control animals (P <0.01 ,P =0.004,P <0.01 ,P <0.01 ,P <0.01 ,respectively).Howerver,SB203580 partly at-tenuated the isoflurane-induced protein change above. Conclusion Isoflurane induces neuroapoptosis in neo-natal rat hippocampus by the activation of p38 MAPK pathway.

Objective To investigate the effects of sevoflurane postconditioning on intestinal ischemiareperfusion (I/R) injury in rats.Methods Thirty-six adult male Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 4 groups (n =9 each):sham operation group (group Sham),group I/R,ischemic postconditioning group (group Ipo) and sevoflurane postconditioning group (group Sevo).Intestinal I/R was induced by clamping the superior mesenteric artery (SMA) for 60 min followed by 120 min of reperfusion in groups I/R,Ipo and Sevo.In group Ipo the animals were subjected to 3 cycles of 30 min reperfusion-30 min ischemia starting from the beginning of reperfusion.The animals inhaled 1.15％ sevoflurane for 30 min starting from the beginning of reperfusion in group Sevo.The animals were sacrificed at 120 min of reperfusion and then the small intestines were removed for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity (by colorimetric method) and caspase-3 protein expression in intestinal tissues (by Western blot).The density of apoptotic cells was calculated by TUNEL.Results Compared with group Sham,the intestinal injury score,density of apoptotic cells and MDA content were significantly increased,SOD activity was decreased,and caspase-3 protein expression was up-regulated in groups I/R,Ipo and Sevo (P ＜ 0.05).Compared with group l/R,the intestinal injury score,density of apoptotic cells and MDA content were significantly decreased,SOD activity was increased,and caspase-3 protein expression was down-regulated in groups Ipo and Sevo (P ＜ 0.05).There was no significant difference in the intestinal injury score,density of apoptotic cells,SOD activity,MDA content and caspase-3 protein expression between Sevo and Ipo groups (P ＞ 0.05).Conclusion Sevoflurane postconditioning can attenuate intestinal I/R injury through reducing lipid peroxidation and cell apoptosis in rats,and the protective effect is similar to that of ischemic post-conditioning.