Objective To investigate the efficiency of transduction of recombinant adenovirus-mediated human endothelial nitric oxide synthase (eNOS) into lung tissue by repeated intratracheal transfection in rats.Methods Sixty 3-4 month old male Wistar rats weighing 220-280 g were randomly divided into 2 groups:control group (group C,n =10) and eNOS gene transduction group (group T,n =50).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 35 mg/kg,tracheally intubated and mechanically ventilated (VT 2.5 ml,RR 60 bpm,FiO2 1.0).Recombinant adenovirus carrying human eNOS gene was given as gift by Professor Gerard from Texas University,Southwest Medical Center.In group T 50 μl of the recombinant adenovirus in concentration of 5 × 109 PFU/ml was instilled into trachea every 5 minutes for 12 times,while in group C equal volume of vector conservation solution was instilled instead.Pulmonary arterial blood samples were obtained at 2,5,7,14 and 21 d after intratracheal transfection (n =10 at each time point) for determination of serum NO concentration.The animals were immediately sacrificed after blood sample collection for determination of expression of eNOS protein in the lung tissue and RNA.The eNOS expression in the trachea,bronchus,lung,liver,spleen and kidney was detected by immuno-histochemistry.Results The serum NO concentrations were significantly higher at all time points in group T than in group C.The eNOS expression was detected in the epithelial cells of trachea and bronchi,and endothelial cells of alveoli and pulmonary blood vessels in group T but not in group C.eNOS expression was not detected in liver,spleen and kidney at 7 d after intratracheal transfection in group T.Conclusion Human eNOS gene mediated by recombinant adenovirus was transducted into rat lung tissue with normal enzyme activity by repeated intratracheal administration without being detected in distant organs.

Objective To explore the pattern of feedback learning deficits in patients with schizophrenia.Methods Twenty-five patients with schizophrenia and 31 controls participated in the study of probabilistic stimulus selection task.The percentage of choose A and avoid B and individual training blocks of reach performance criterion were analyzed.Results There was no significant difference on the percentage ofchoose A between patient group and control group(control group:(66.13±26.31) ％ ; patient group:(63.75±20.57) ％ ; t=0.37,P=0.713).The percentage of avoid B in patient group was significantly lower than that in control group(control group:(62.10±27.10)％;patient group:(49.75±13.68)％; t=2.212,P=0.032).In addition,the training blocks of reach performance criterion in patient group was significantly greater than that in control group (control group:3.23±2.012;patient group:4.64±1.977; t=-2.635,P=0.011).Conclusion The deficits of feedback learning in patients with schizophrenia is largely due to the failure of avoiding negative feedback stimuli.Learning efficiency was lower in patients with schizophrenia than controls.

Objective To investigate the effects of penehyclidine hydrochloride on blood-brain barrier in a rat model of cardiopulmonary bypass ( CPB) . Methods Sixty adult male SD rats, aged 4-6 months, weighing 320- 370 g, were randomly divided into 5 groups ( n = 12 each) : sham operation group (group S), CPB group, and low-, median- and high-dose penehyclidine hydrochloride groups (groups LP, MP and HP). The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg, intubated and mechanically ventilated. The femoral and jugular arteries and jugular vein were cannulated. CPB was performed for 60 min. Penehyclidine hydrochloride 0.2, 0.6 and 2.0 mg/kg were added to the priming solution in groups LP, MP and HP respectively, while the equal volume of normal saline was added in group CPB. Evans blue was injected via femoral vein at 1 h before the animals were sacrificed. Six rats in each group were sacrificed, their brains immediately removed and the hippocampi isolated for determination of Evans blue content. The other rats were sacrificed and the hippocampi isolated to determine the water content and observe the ultrastructure of blood-brain barrier. Results Compared with group S, the Evans blue content and water content were significantly increased in the other groups ( P ＜ 0.05) . Compared with groups CPB and LP, the Evans blue content and water content were significantly decreased in groups MP and HP ( P ＜ 0.05) . The Evans blue content was significantly lower in group HP than in group MP ( P ＜ 0.05). The CPB-induced changes were significantly attenuated in groups MP and HP compared with groups CPB and LP. Conclusion Penehyclidine hydrochloride can protect blood-brain barrier against the CPB-induced injury and the effect is related to the dose.

ObjectiveTo investigate the effect of penehyclidine hydrochloride on acute lung injury induced by cardiopulmonary bypass (CPB) in rats.MethodsForty adult male SD rats aged 4-6 months weighing 330-420 g were randomly divided into4 groups ( n =10 each): sham operation group (group S),acute lung injury group (group ALI) and low and high dose of penehyclidine hydrochloride groups (groups PL and PH ).Penehyclidine hydrochloride 0.6 and 2.0 mg/kg were added to the priming solution in groups PL and PH,while the equal volume of normal saline was added in group ALI instead.The rats of groups ALI,PL and PH were underwent 1 h of CPB.Arterial blood samples were collected before CPB and at 2 h after CPB for blood gas analysis.The superior vera cava blood samples and lung tissues were collected at 2 h after CPB for determination of concentrations of TNF-α and IL-6,lung tissue contents of water and malondialdehyde (MDA) and activity of glutathione peroxidase (GSH-px).The pathological change of lung tissue was also examined.ResultsCompared with group S,PaO2 was significantly decreased at 2 h after CPB,plasma concentrations of TNF-α and IL-6 and contents of water and MDA in lung tissues were increased,while activity of GSH-px in lung tissues was decreased in groups ALI,PL and PH ( R ＜ 0.05).Compared with group ALI,PaO2 was significantly increased at 2 h after CPB,plasma concentrations of TNF-α and IL-6 and contents of water and MDA in lung tissues were decreased,activity of GSH-px in lung tissues was increased (P ＜ 0.05),and the pathological change was reduced in groups PL and PH.Compared with group PL,PaO2 was significantly increased at 2 h after CPB,plasma concentrations of TNF-α and IL-6 and contents of water and MDA in lung tissues were decreased,activity of GSH-px in lung tissues was increased ( P ＜0.05),and the pathological change was reduced more obviously in group PH.ConclusionPenehyclidine hydrochloride 0.6 or 2.0 mg/kg can reduce the CPB-induced lung injury in a dose-dependent manner by antioxidant and anti-inflammatory mechanism in rats.

Objective To evaluate the effect of penehyclidine hydrochloride (PHC) on the level of angiopoietin-1 (Ang-1) and tyrosine kinase receptor-2 (Tie-2) during endotoxin-induced acute lung injury (ALI) in rats.Methods Forty adult male Sprague-Dawley rats,weighing 180-220 g,were randomly divided into 4 groups using a random number table (n =10 each):control group (group C),ALI group,low-dose PHC group (group L-PHC) and high-dose PHC group (group H-PHC).ALI was induced with iv injection of lipopolysaccharide 5.0 mg/kg via the tail vein.In L-PHC and H-PHC groups,PHC 0.6 and 2 mg/kg were injected,respectively,via the tail vein at 1 and 24 h after lipopolysaccharide injection.The rats were sacrificed at 48 h after the initial injection of PHC to measure the lung water content,protein concentration in bronchoalveolar lavage fluid (BALF),and the expression of Ang-1,Tie-2 and phosphorylated Tie-2 in lung tissues.The morphological changes of lung tissues were observed under light microscope and the ultrastructural changes of alveolar epithelial barrier under transmission electron microscope.Results Compared with group C,the lung water content and protein concentrations in BALF were significantly increased,and the expression of Ang-1 and phosphorylated Tie-2 was down-regulated in the other three groups (P ＜ 0.05).Compared with group ALI,the lung water content and protein concentrations in BALF were significantly decreased,and the expression of Ang-1 and phosphorylated Tie-2 was up-regulated in H-PHC group (P ＜ 0.05),and no significant changes were found in the parameters mentioned above in group L-PHC (P ＞0.05).The damage to lung tissues was significantly reduced in group H-PHC as compared with group ALI.Conclusion PHC can improve the permeability of pulmonary microvascular and reduce injury to alveolar epithelial barrier,thus ameliorating endotoxin-induced ALI in rats,and the effect is dose-related and up-regulation of Ang-1 expression and inhancement of Tie-2 activity are involved in the mechanism.

Objective To investigate the effect of penehyclidine hydrochloride on brain injury induced by cardiopulmonary bypass (CPB) in rats. Methods Thirty adult male SD rats were randomly divided into 5 groups ( n = 6 each): sham operation group (group S), CPB group, and low, median and high dose penehyclidine hydrochloride groups (group PL, PM , PH). Penehyclidine hydrochloride 0.2, 0.6 and 2.0 mg/kg were added to the priming solution in group PL, PM and PH respectively, while the equal volume of normal saline was added instead in group S. Blood samples were obtained at 2 h after termination of CPB to determine the plasma concentrations of neuron specific enolase (NSE) and S-100β protein. The brain tissues were taken to observe the ultrastructure of hippocampal neurons with electron microscope. Results The concentrations of NSE and S-100β protein were significantly higher in the other groups than in group S, while lower in group PM and PH than in group CPB and PL( P＜ 0.05). The S-100β protein concentration was significantly lower in group PH than in group PM( P ＜ 0.05). The damage to hippocampal neurons was significantly attenuated in group PM and Ps. Conclusion Penehyclidine hydrochloride 0.6 or 2.0 mg/kg can reduce the CPB-induced brain injury in a dose-dependent manner in rats.

Objective To investigate the effects of cardiopulmonary bypass (CPB) on the expression of tight junction protein occludin in rat lung tissues.Methods Twenty adult male Sprague-Dawley rats were randomly divided into 2 groups (n=10 each): sham operation group (group S) and CPBgroup.The rats underwent 1 h of CPB and were observed 2 h later in group CPB.The lung water content,neutrophil percentage and protein concentration in bronchoalveolar lavage fluid (BALF) were measured.The expression of occludin in lung tissues was detected by immunohistochemistry and Western-blot analysis.The ultrastructure of alveolar epithelial barrier was observed with transmission electron microscope.Results Compared with group S,the lung water content,protein concentration in BALF and neutrophil percentage were significantly increased (P ＜ 0.05),the expression of occludin in lung tissues was significantly down-regulated (P ＜ 0.05) and the damage to alveolar epithelial barrier was aggravated in group CPB.Conclusion The expression of occludin in lung tissues is down-regulated and the damage to alveolar epithelial barrier is induced after CPB,which may be one of the important factors in acute lung injury induced by CPB.

Objective To evaluate the effect of hemorrhagic shock factor on the pharmacokinetics of rocuronium in pigs.Methods Sixteen pathogen-free Bama miniature pigs of both sexes,aged 3-5 months,weighing 22-25 kg,were divided into 2 groups (n=8 each) using a random number table:control group (group C) and hemorrhagic shock group (group HS).In group C,rocuronium 3.78 mg/kg was injected via the auricular vein.In group HS,the animals were subjected to volume-controlled hemorrhage,about 40％ of blood volume was withdrawn from the left femoral artery over 15 min (30 ml/kg),and rocuronium 3.78 mg/kg was injected via the auricular vein after the model was successfully established.At 0,2,4,7,10,15,20,30,60,120,180,240,300,360 and 420 min after rocuronium injection,blood samples were collected from the internal jugular vein for determination of the plasma concentration of rocuronium by high-performance liquid chromatography-tandem mass spectrometry.The pharmacokinetic parameters of rocuronium were calculated.Results Compared with group C,the plasma concentration of rocuronium was significantly increased at 20 and 60-420 min after rocuronium injection,the elimination half-life and mean residence time were prolonged,and the plasma effect-site equilibration rate constant was decreased in group HS (P＜0.05).There was no significant difference in the maximal concentration and area under the concentration-time curve between the two groups (P＞ 0.05).Conclusion The elimination of rocuronium is slower in a pig model of hemorrhagic shock.

Objective To evaluate the effect of dexmedetomidine on acute liver injury in rats with endotoxemia.Methods Eighteen adult male Sprague-Dawley rats, aged 3-4 months, weighing 250-300 g, were randomly divided into 3 groups (n=6 each) using a random number table: control group (group C), endotoxin group (group E), and dexmedetomidine group (group D).In E and D groups, lipopo-lysaccharide 5 mg/kg was injected via the femoral vein of rats anesthetized with chloral hydrate.In group D, dexmedetomidine was infused with a 7 μg/kg loading bolus over 15 min after injection of lipopolysaccharide, followed by a 6 h continuous infusion of 5 μg · kg-1 · h-1.The equal volume of normal saline was given instead in E and C groups.After the end of administration, blood samples from the femoral vein were drawn for determination of tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) concentrations in serum (by using enzyme-linked immunosorbent assay), and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum (using the International Federation of Clinical Chemistry and Laboratory Medicine reference procedures).Liver specimens were obtained for examination of pathologic changes with electron microscope.Results Compared with group C, the serum ALT and AST activities and TNF-α and IL-lβ concentrations were significantly increased in E and D groups.Compared with group E, the serum ALT and AST activities and TNF-α and IL-1β concentrations were significantly decreased in group D.The pathologic changes of livers were obvious in group E, and were significantly reduced in group D compared with group E.Conclusion Dexmedetomidine can alleviate acute liver injury in rats with endotoxemia, and the underlying mechanism is associated with inhibition of systemic inflammatory responses.

Objective To investigate the effect of dexmedetomidine on acute kidney injury in endotoxemic rats.Methods Thirty adult male Sprague-Dawley rats,aged 4-6 months,weighing 180-220 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group (group C),lipopolysaccharide group (group L),and dexmedetomidine (group D).Lipopolysaccharide (LPS) 5 mg/kg was injected slowly into the femoral vein to establish the model of endotoxemic in rats anesthetized with chloral hydrate.In group D,after LPS injection,a loading dose of dexmedetomidine 7 μg/kg was injected intravenously,and 15 min later dexmedetomidine was infused for 6 h at 5 μg · kg-1 · h-1,while the equal volume of normal saline was given in L and C groups.At 6 h after the end of LPS administration,blood samples were collected from the femoral vein for determination of serum creatinine (Cr),blood urea nitrogen (BUN),tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) concentrations.At 24 h after the end of LPS administration,the animals were sacrificed and kidneys were removed for microscopic examination and for determination of the expression of tight junction proteins ZO-1 and occludin in renal tissues by Western blot.Results Compared with group C,the serum Cr,BUN,TNF-α and IL-6 concentrations were significantly increased,and the expression of ZO-1 and occluding was down-regulated in L and D groups.Compared with group L,the serum Cr,BUN,TNF-α and IL-6 concentrations were significantly decreased,the expression of ZO-1 and occluding was up-regulated,and the pathological changes of kidneys were mitigated in D group.Conclusion Dexmedetomidine can alleviate acute kidney injury in endotoxemic rats.