Rat liver mitochondria were exposed to various conentrations of halothane,enflurane, isoflurane and sevoflurane. Electron transfer rates from NADH and succinate to cytochrome C were measured by scanning dual wavelength spectrophotometer. Statistical analysis of the data suggested that halothane at clinical or higher than clinical concentrations markedly inhibited activities of NADH-Cyt,C reductase.in contrast,no decrease occurred in the activities of NADH dehydrogenase,NADH-coenzyme Q reductase and enzymatical system of succinate chain. Enflurane,isoflurane and sevoflurane had little effect on enzymatical system of mitochondrial electron transfer chain. These data indicate that halothane interfere with utilization of NADH-linked substrate by blocking electon transport from NADH to cytochrome C and it is probable that the locus of action is at Q binding protein(Qpn) or complex of Qpn and ubiquinone.

Objective In order to improve the survival of graft liver after liver transplantation, this study was designed to investigate whether intraportal injection of 150mg/kg N-acetylcysteine (NAC) in rats could reduce hepatic ischemia-reperfusion injury after 48 h of cold storage and 2 h of reperfusion.Methods Healthy male Wistar rats weighing 250-350g were used.The study consisted of three groups: control group (group Ⅰ);NAC-treated group(group Ⅱ).1 ml of 5% dextrose (D5%) or 1 ml D5% containing 150mg/kg NAC was injected into the superior mesenteric vein.15 min after the injection of D5% or NAC the liver was flushed with cold (4℃) Ringer's solution through the portal vein .After perfusion, the liver was removed and kept in 100 ml UW solution at 4℃ for 48 h.In group Ⅲ animals were pretreated with buthionine sulfoximine (BSO) 2 h before intraportal injection of D5% or NAC and liver harvesting.After cold storage, the livers were then perfused for 2 h by a closed circulating system.Aspartate transaminase (AST), alanine transaminase (ALT), and lactate dehydrogenase (LDH) activities in the perfusate were determined by reflectometry.Lactate and acid phosphatase activities were determined by enzymatic methods.Results After 48 h of cold storage and 2 h of reperfusion, livers from NAC-treated group produced larger amounts of bile than those in the control group, and released less LDH, AST, ALT and acid phosphatase, a marker of Kupffer cell injury in the perfusate.The protective effects of NAC against cold ischemia-reperfusion liver injury were maintained when animals were pretreated with BSO, a specific inhibitor of glutathione synthesis.Conclusions This study shows that intraportal administration of NAC in vivo significantly improves the initial function of the isolated rat liver.Our results also indicate that NAC inhibits the activation of Kupffer cells, which are the first source of reactive oxygen intermediates during reperfusion.

ObjectiveIn order to improve the survival of graft liver after liver transplantation, this study was designed to investigate whether intraportal injection of 150mg/kg N-acetylcysteine (NAC) in rats could reduce hepatic ischemia-reperfusion injury after 48 h of cold storage and 2 h of reperfusion. Methods Healthy male Wistar rats weighing 250-350g were used. The study consisted of three groups: control group (group Ⅰ) ;NAC-treated group(group Ⅱ). 1 ml of 5% dextrose (D5%) or 1 ml D5% containing 150mg/ kg NAC was injected into the superior mesenteric vein. 15 min after the injection of D5 % or NAC the liver was flushed with cold (4℃) Ringer' s solution through the portal vein . After perfusion, the liver was removed and kept in 100 ml UW solution at 4℃ for 48 h. In group Ⅲ animals were pretreated with buthionine sulfoximine (BSO) 2 h before intraportal injection of D5 % or NAC and liver harvesting. After cold storage, the livers were then perfused for 2 h by a closed circulating system. Aspartate transaminase (AST), alanine transaminase (ALT), and lactate dehydrogenase (LDH) activities in the perfusate were determined by reflectometry. Lactate and acid phosphatase activities were determined by enzymatic methods. ResultsAfter 48 h of cold storage and 2 h of reperfusion, livers from NAC-treated group produced larger amounts of bile than those in the control group, and released less LDH, AST, ALT and acid phosphatase, a marker of Kupffer cell injury in the perfusate. The protective effects of NAC against cold ischemia-reperfusion liver injury were maintained when animals were pretreated with BSO, a specific inhibitor of glutathione synthesis. ConclusionsThis study shows that intraportal administration of NAC in vivo significantly improves the initial function of the isolated rat liver. Our results also indicate that NAC inhibits the activation of Kupffer cells, which are the first source of reactive oxygen intermediates during reperfusion.

Objective To establish a method for acute isolation of rat superior cervical ganglion (SCG) cells and identify the electrophysiological properties.Methods Sprague-Dawley rats of both sexes,aged 5-12 days,were decapitated.The SCGs were removed quickly,and the single SCG cell was enzymatically isolated from the SCGs.When the holding potential was - 60 mV,100 μmol/L acetylcholine was applied and the nicotinic acetylcholine receptor currents were recorded by whole-cell patch-clamp technique.When the holding potential was 760 mV,65 mmol/L KCl was applied and quantal release of catecholamines was detected by using carbon fiber electrodes.Results SCG cells with normal electrophysiological properties were isolated.Typical nicotinic acetylcholine receptor currents and quantal release of catecholamines were recorded successfully.Conclusion The cells suitable for patch-clamp experiments can be obtained by using the method for acute isolation of rat SCG cells.

The use of genetic approaches to probe relative genes that control sensitivity to volatile anesthetics in intact model has recently emerged as the powerful tools and strategies in dissecting mechanisms of anesthesia. Multiple model organisms such as yeast, nematodes, fruitflies and mammals are currently being exploited, and a number of sensitive genes have been screened, with some of them being cloned, located, and function identified. The emerging technologies are likely to provide further great advances for elucidating the specific anesthetic molecular sites.

G1.(3)Among 5 groups,the highest incidences of nausea,vomiting and pruritis were observed in group G1 for about 24% 32%,but with no statistical difference compared with other groups.Conclusion: With equal doses of bupivacaine and fentanyl mixed,the concentration/volume ratios may affect the analgesic effects of postoperative epidural analgesia in patients with hepatectomy.

Clinical practice is an important teaching stage for interns of anesthesiology in medical universities.We discuss advantages and disadvantages for different teaching modes in the course of clinical practice,aiming at choosing a better one to optimize practice quality and bringing up graduates with high diathesis.

Objective To investigate the effect of different concentrations of heparin in the cold preservation solution on blood coagulation after reperfusion during orthotopic liver transplantation ( OLT) . Methods Forty ASA Ⅱ or Ⅲ patients with end-stage liver disease (31 males, 9 females) weighing 51-82 kg undergoing OLT were randomly divided into 3 groups according to the concentrations of heparin in the liver preservation solution: group Ⅰ 2 000 U?L-1( n = 14); group Ⅱ 8000 U?L-1(n= 12) and group Ⅲ 20 000 U?L-1(n = 12) .Anesthesia was induced with propofol, fentanyl and rocuronium and maintained with isoflurane inhalation and intermittent i.v. boluses of fentanyl. Body temperature was maintained at 35.5-37.5℃ and Hct at 22%-35% . Blood samples were taken from central vein at 15 min before reperfusion (T0,baseline) and 5, 30, 60, 120 and 180 min of reperfusion (T1-5) for determination of ACT, CR and platelet function (PF) using Sonoclot platelet function and coagulation analyzer (U.S.A.). Blood concentration of heparin was determined in 3-4 patients in each group at T0-5 ( HPLC) . Results The 3 groups were comparable with respect to age, body weight, ASA physical status and Child classification. The glass ball activated coagulation time (gbACT) was significantly prolonged at T1-4 in group Ⅲ and at T1,2 in group Ⅱ as compared to the baseline values at T0 ( P

Objective To determine whether thiol can affect the activity of nuclear factor kappa B (NF-?B) in and release of TNF-?from lipopolysaccharide (LPS)-activated kupffer cells (KCs) and ascertain whether these effects are mediated through glutathione (GSH) .Methods KCs were isolated from livers of healthy male adult SD rats weighing 250-300g and cultured for 12h. The cultured KCs were exposed to LPS 10 ng?ml-1. The effects of different concentrations of glutathione monoethylester (GSHEE) and N-acetylcysteine (NAC) on NF-?B activity in and release of TNF-afrom KCs were determined. The effect of pretreatment with GSH synthesis inhibitor - BSO on the inhibitory effect of NAC on inflammation was investigated. NF?B activity was determined by electrophoretic mobility shift assay (EMSA) and intracellular GSH content and TNF-?level in supernatant by HPLC. Results Intracellular GSH content remained unchanged after exposure of KCs to LPS. Both GSHEE and NAC increased intracellular GSH level and inhibited NF-?B activity and release of TNF-?. BSO blocked the increase in GSH induced by NAC but did not affect the inhibitory effect of NAC on TNF?release. Conclusions LPS does not affect GSH level in KCs. NAC can inhibit NF-?B activity in and TNF-?release from KCs and increase intracellular GSH level but the inhibitory effect is not mediated through GSH.