Objective To investigate the effects of pretreatment with Chinese herbal Sini Decoction ( SND) on the acute lung injury induced by intestinal ischemia / reperfusion (I/R) .Methods Thirty-two healthy SD rats of both sexes weighing 275-300 g were randomly divided into four groups of 8 animals : (Ⅰ) control group in which sham operation was performed, ( Ⅱ) I/R group in which superior mesenteric artery was clamped for 1 h followed by 3 h reperfusion; (Ⅲ) and (Ⅳ) SND group 1 and 2 in which SND 3 g?kg-1 ( Ⅲ) or 6 g?kg-1 (Ⅳ) was given via gastric tube every day for 3 days before I/R. Carotid artery was cannulated for MAP monitoring. The animals were sacrificed by decapitation at the end of 3 h reperfusion. Blood was collected and the lungs were immediately removed for determination of lung water content [ (wet weight - dry weight) / wet weight ?100% ], lung NO, endothelin-1 (ET-1) and MDA contents and SOD activity, lung permeability index (BALF protein concentration/serum protein concentration) and microscopic examination. Results SND pretreatment significantly alleviated the hypotension and morphological changes of the lungs induced by intestinal I/R. Lung water content, lung permeability index and lung MDA and NO contents increased significantly whereas lung SOD activity significantly decreased in I/R group ( group Ⅱ) compared with those in control group ( P

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 effects of electrical stimulation of vagus nerve on gut injury following intestinal ischemia-reperfusion in rats. METHODS: 30 adult male Wistar rats subjected to bilateral cervical vagotomy were randomly divided into three groups (n=10 per group): (1) Intestinal ischemia-reperfusion group (group I/R): laparotomy and I/R induced by clamping arteria mesenterica superior for 1 h followed by reperfusion for 2 h. (2) Vagus nerve stimulation group (group VNS): laparotomy, I/R and electric stimulation with pulse train of constant amplitude 5V, pulse width 2 ms and frequency 1 Hz at the left caudal vagus ends for 20 minutes before and after occlusion. (3) Sham control group (group SC): sham operation and sham stimulation. Carotid artery was cannulated for mean arterial pressure (MAP) monitoring. A strip of small intestine was taken from distal end of ileum for light microscopic (LM) and transient electron microscopic (TEM) examination at the time of 2 h after reperfusion. Improved Chiu’s scale was used to quantitatively assay the damage degree. The levels of malondialdehyde (MDA) and TNF-? in plasma were detected. RESULTS: MAP in every group kept steady during ischemia, but decreased gradually with the prolongation in the time of reperfusion. MAP decreased more dramaticly in group I/R than that in group VNS (P

Objective To investigate the effect of isoflurane on the spatial learning and memory in aged mice,and whether this is associated with the changes of Caspase-3 expression and apoptosis in brain.Methods Twenty-four CO57BL/6 aged mice(16 months)were randomly divided into isoflurane treatment group(Iso Group,n=12) and sham control group (Con Group,n=12).Mice in Iso group were exposed to 1% isoflurane in carrying gas of 30% oxygen,balance nitrogen in a warmed,humidified chamber for4 h per day for2 days.For Con group,animals were treated at the same condition with only carrying gas.After anesthetic exposures,behavioral testing was performed using the Morris water maze(MWM),and then changes of Caspase-3 expression and apoptosis in hippocampus CAI,dentate gyrus(DG) and cortex(CX) in brain were determined by using immunofluorecence staining and TUNEL staining.Results In hidden-platform training of MWM,the mean escape latency to platform showed no significant difference between the two groups (F=0.007,P=1.235),but the mice in Iso group showed obviously impaired retention of memory by spending more percentage of time swimming in the probe quadrant as compared to the control animals in the probe test((34.5±5.0)%vs(45.1±4.9)%.P<0.01).There was no significant difference in average swimming speed during the MWM testing trials between the two groups (F=1.537,P=0.241).A few Caspase-3 and apoptosis positive cells were found in hippocampus Cal,DG and CX regious,while no difference was found in the density of positive cells between the two groups(P>0.05).Conclusion 1% isoflurane repeatedly exposure significantly impaires the spatial reference memory in aged mouses,however does not significantly change the expression of easpase-3 and apoptosis in brain.

Objective To determine whether fetal rats exposure to isoflurane will cause postnatal learning and memory deficits,and change Bcl-2/Bax ratio in the hippocampus CA1 and retrosplenial cortex in fetal brain of rats. Methods Twenty-eight Sprague Dawley pregnant rats at gestation day 21 (E21) were randomly divided into isoflurane treatment group(n=14) and sham control group(n=14). Rats in isoflurane treatment group were ex-posed to 1.3% isoflurane in a carrying gas of 30% oxygen, balance nitrogen for 6 h in a warmed, humidified cham-ber. For sham control group,animals were treated at the same condition with only carrying gas. In behavior study,the spatial learning and memory ability at juvenile ages was determined with the Morris Water Maze(MWM). In immunohistochemistry study,changes of Bcl-2 and Bax expression in hippocampus CA1 and retrosplenial cortex in the fetus brain after isoflurane treatment at 2 hours was performed by using immunofluorecence staining.Results In the MWM training, the escape latency to platform in the place trials showed no significant difference between the two groups,but the postnatal rats in 1.3% isoflurane group showed obviously improved retention of memory by spending more percentage of time swimming in the probe quadrant as compared to the control animals ((42.33±2.31) s vs (33.2±2.15) s, t=2.21, P<0.05) in the probe test. Compared to controls, 1.3% isoflu-rane exposure for 6 h to the pregnant rats increased the intensity of Bcl-2, decreased the intensity of Bax, and sig-nificantly increased the Bcl-2/Bax ratio in the fetal hippocampal CA1 region (4.40±0. 86 vs 1.31±0.32, t=3. 378, P<0.01) and the fetal retrosplenial cortex (5.07±1.27 vs 1.47±0.48, t=2.656, P < 0.05) respec-tively. Conclusion 1.3% isoflurane exposure in pregnant rats significantly improves the spatial retention memo-ry of their rat pups at a juvenile age and increases the Bcl-2/Bax ratio in the hippocampal CA1 region and the ret-resplenial cortex in the fetal rat brains.

Objective To investigate the practical value of Shikani optical stylet used for orotracheal intubation in critically conscious patients and to evaluate the successful rate. Method A total of 48 conscious patients with severe respiratory failure were selected from January 2008 to June 2009. Of them, there were 31 males and 17 females,aging 21-86 years old with an average of 57 years old. All enrolled patients needed endotracheal intubation for mechanical ventilation support, and they were assigned to Shikani group (group S, n = 25) and Macintosh group (group M, n = 23) according to the odd and even number of date of admission to this study. The time consumed for intubation, the number of failure in intubation, the adverse effects or complications such as hemodynam-ic changes, injury to the pharyngo-oral cavity, choking and breath-hold were observed and recorded. The rank test and chi-square test were used for statistical analysis. Results The ratio of the successful intubation at first attempt was much higher in group S (96.0%) than that in group M (60.9%) (P <0.01). Compared with group M, the time consumed for intubation was significantly shortened, the cardiovascular reactions were much mild, and the incidence of injury to pharyngo-oral cavity, choking and breath-hold were less in the group S ( P < 0.01 all). Conclusions For the acute and critical patients, especially the conscious ones, orotracheal intubation with the Shikani optical stylet is rapid, successful, safe and less injurious, resulting in mild cardiovascular reactions.

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.

Objective To investigate the effects of isoflurane and sevoflurane at the equivalent depth of subanesthesia on neuronal proliferation and phosphorylation of extraceullar signal-regulated kinase 1/2 (ERK1/2)protein in the hippocampi of neonatal rats.Methods Seventy-two neonatal rats at postnatal day 7 were involved in this study and they were assigned randomly into isoflurane group (Iso group),sevoflurane group (Sev group) and control group (Con group).The rats in I group,S group or C group were separately exposed to 0.75％ isoflurane or 1.2％ sevoflurane (equivalent to 0.3 MAC for neonatal rats) or air for 6 h.Some rats in each group were injected intraperitoneally BrdU 100 mg/kg immediately (D0) (n =6) or 3 days after exposure (D3) (n =6),and their brains were perfusion and embedded by paraffin 24 h after BrdU injection.BrdU positive expressions in the in dentate gyrus (DG) area of hippocampus were detected by IHC staining.Besides,the fresh hippocampi of some rats each group were dissected at the end of anesthesia,caspase-3 and phospho-ERK1/2,ERK1/2 proteins expression were detected by Western blot (n =6).The other rats in each group were used to measure changes of pH and blood glucose (n =6).One way ANOVA test was used for data analysis among groups.Results BrdU-positive cells had no significant difference among group IsoD0 ((1332.43 ± 192.70)/mm2),group SevD0 ((1207.33 ±139.50)/mm2),and group ConDO ((1362.40 ± 227.90)/mm2) at D0,while which had significantly decreased by 32.6％ (P＜ 0.05) in group IsoD3 ((604.56 ± 65.77)/mm2) when compared with those in group ConD3 ((896.90 ± 78.77)/mm2) at D3.There was no significant difference between groups of SevD3 ((808.73 ± 41.27)/mm2) and ConD3.The expression of caspase-3 protein was increased by 195％ (P＜ 0.01) in Iso group while which only increased by 74％ (P ＜ 0.05) in Sev group when compared with Con group.The expression of P44 and P42 of phospho-ERK1/2 protein in the hippocami decreased by 53％ (P ＜ 0.01) and 47％ (P ＜ 0.01))seperately in Iso group when compared with Con group,while there were no significant differences between Sev group and Con group.Conclusion 0.3 MAC isoflurane,not sevoflurane inhibits neuronal proliferation in DG of hippocampi in the neonatal rats.Inhibiting ERK1/2 phosphorylation may involve in the mechanisms of that isoflurane inhibits neuronal proliferation.

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.