Objective To investigate whether diazoxide preconditioning can exert protective effect on myocardium against ischemia-reperfusion injury in immature rabbits and the possible mechanism. Methods Twenty-one healthy 3-4 week old white rabbits of either sex were randomly divided into 3 groups : group Ⅰ control ( n = 8) ; group Ⅱ diazoxide preconditioning ( n = 8) and group Ⅲ diazoxide + 5-HD preconditioning ( n = 5) . The animals were anesthetized with intraperitoneal pentobarbital 50 mg?kg-1 and heparized (150 IU?kg-1). The hearts were excised and connected to Langendorff apparatus and passively perfused with normothermic (37℃), oxygenated (95% O2 , 5% CO2) Krebs-Henseleit buffer (KHB) at a constant perfusion pressure of 70cmH2O. A latex balloon was inserted via left atrium into left ventricle and filled with water. The left ventricular end-diastolic pressure (LVEDP) was maintained at 10 mm Hg. In group I cardiac arrest was induced with St Thomas Ⅱ solution after the heart was perfused with KHB for 30 min. In group 11 after being perfused with KHB for 15 min, the hearts were perfused with diazoxide 100?mol?L-1 for 5 min followed by 10 min wash-out with KHB , then cardiac arrest was induced as in group Ⅰ . In group Ⅲ after being perfused with KHB for 15 min, the hearts were perfused with diazoxide 100?mol?L-1 and 5-HD 100?mol?L-1 for 5 min, followed by 10 min wash-out with KHB, then the cardiac arrest was induced as in group Ⅰ and Ⅱ . All hearts were subjected to 30 min ischemia followed by 45 min reperfusion after cardiac arrest. Coronary flow (CF), HR, left ventricular developed pressure (LVDP) and dp / dt max were measured after the hearts were perfused with KHB for 15 min (baseline) and at 5, 10, 15, 30, 45 min after reperfusion was resumed. Coronary effluent was collected at 5 min after reperfusion was resumed for determination of myocardial enzymes, CK, LDH and AST. At the end of experiment myocardial specimen was obtained for determination of ATP content and ultrastructure examination. Results There was no significant difference in the baseline hemodynamic parameters among the three groups. The rates of recovery of LVDP and ? dp / dt max after reperfusion were significantly higher in group Ⅱ than those in group I and Ⅲ ( P 0.05 ) , Conclusion Diazoxide can protect myocardium from ischemia-reperfusion injury by opening the mitochondrial KATP channel in immature rabbits.

We analyzed the special characteristics of postgraduate education for anesthesiology majors based on the current situation of medical postgraduate education.Next,we introduced the ‘ sandwich' teaching method and discussed how it can promote students' subjective initiative by transforming various learning patterns.Finally,the effectiveness of study was elevated by ‘sandwich' teaching method.

ObjectiveTo investigate the effect of cardiopulmonary bypass (CPB) on the secretory function of islet cells in rabbits.MethodsTwenty adult New Zealand white rabbits of both sexes,weighing 2.5-3.0kg,were randomly divided into 2 groups ( n =10 each):sham operation group (group S) and CPB group.The rabbits were anesthetized with 3％ pentobarbital sodium 30 mg/kg.Blood samples were collected from the left femoral artery at 5 min after anesthesia (T1),immediately before CPB (T2 ),immediately after aortic clamping (T3 ),and at 5,35 and 75 min after aortic unclamping (T4-6) in the two groups for determination of levels of blood glucose,insulin and glucagons.Insulin resistance index was calculated.ResultsCompared with group S,the blood glucose concentration and levels of insulin and glucagons and insulin resistance index at T3-6 were significantly increased in group CPB ( P ＜ 0.05).ConclusionAlthough increase in blood glucose enhances the secretion of insulin in islet β cells,hyperglycemia cannot be compensated completely by the increased insulin during CPB in rabbits.The increase in blood glucose may be related to islet α cell resistance.

Objective To investigate the effect of Shenfu injectio on insulin resistance in rabbits undergoing cardiopulmonary bypass (CPB) . Methods Thirty New Zealand white rabbits of both sexes weighing 2.2-2.5 kg were randomly divided into 3 groups (n = 10 each): group I sham operation (group S); group Ⅰ CPB and group Ⅰ CPB + Shenfu injectio (group SFI). In group IE (group SFI) Shenfii injectio 10 ml/kg was injected intraperitoneally (IP) once a day for 2 days and at 30 min before anesthesia. In group Ⅱ and Ⅲ the duration between aortic cross-clamping and unclamping was 60 min. MAP was maintained at 50-60 mm Hg during CPB. The plasma glucose and insulin concentrations were measured immediately after induction of anesthesia (T_1), immediately after aortic cross-clamping (T_2) , and at 5, 35 and 75 min after aortic unclamping (T_3, T_4 , T_5) and insulin resistance index (HOMA-IR) was calculated. The expression of insulin receptor substrate-1 (IRS-1), phosphatidyl-inositol 3 kinase (PI3Kp85) and glucose transporters (CLUT4) in skeletal muscle was detected at 150 min of CPB by immuno-histochemistry. Results Compared with the baseline values at T_1 , plasma glucose and insulin concentrations and HOMA-IR were significantly increased at T_(2-5) in all 3 groups ( P < 0.05) . CPB significantly increased the plasma glucose and insulin concentrations and HOMA-IR at T_(2-5) in group Ⅱ and Ⅲ as compared with group S ( P < 0.05). The expression of PI3Kp85 and GLUT4 in muscle was significantly decreased in group Ⅱ and Ⅲ as compared with group S ( P < 0.05). Shenfu injectio significantly decreased plasma glucose concentration and HOMA-IR and increased plasma insulin concentration in group fD as compared with group Ⅱ . Shenfu injectio significantly attenuated the CPB-induced changes in IRS-1, PI3Kp85 and GLUT4 expression in muscle. Conclusion Shenfu injectio can ameliorate CPB-induced insulin resistance and the mechanism may be related to the up-regulation of the expression of insulin signal transduction molecules in skeletal muscle.

Objective To investigate the blood-saving effect of controlled low central venous pressure (CLCVP) in different types of hepatectomy.Methods Ninety ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 37-76 yr,weighing 40-75 kg,undergoing elective hepatectomy,were divided into 6 groups according to the surgical approach and whether CLCVP was used during surgery (n =15 each):CLCVP1-3 groups and nonCLCVP1-3 groups (NCLCVP1-3 groups).The standard hepatectomy,half liver resection and irregular hepatectomy were performed in CLCVP1-3 groups,respectively,with CLCVP.The standard hepatectomy,half liver resection and irregular hepatectomy were performed in NCLCVP1-3 groups,respectively,without CLCVP.In CLCVP1-3 groups,from skin incision to the end of liver resection,CVP was maintained ≤ 5 cm H2 O through adjustment of the position,fluid restriction and iv infusion of nitroglycerin,and norepinephrine was infused simultaneously to maintain mean arterial pressure ≥ 60 mm Hg.In NCLCVP1-3 groups CVP was maintained at 6-12 cm H2O.Intraoperative blood loss and blood transfusion were recorded.Results Compared with NCLCVP1-3 groups,intraoperative blood loss was significantly decreased in CLCVP1-3 groups (P ＜ 0.05).Compared with NCLCVP3 group,the amount of blood transfusion was significantly decreased,the constituent ratio of intraoperative blood loss ＜ 200 ml was increased,and the constituent ratio of intraoperative blood loss ＞ 1000 ml was decreased in group CLCVP3 (P ＜ 0.05).Conclusion CLCVP can decrease the intraoperative blood loss and blood transfusion in patients undergoing irregular hepatectomy.

Objective To evaluate the effect of pretreatment with nerve growth factor-beta (NGF-β) on ischemia-reperfusion (I/R) injury in isolated rat hearts by comparing it with NGF-β preconditioning.Methods Pathogen-free male Sprague-Dawley rats,weighing 200-300 g,aged 8-10 weeks,were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with KH solution aerated with 95％ O2 and 5％ CO2 at 37℃.Twenty-four isolated rat hearts were randomly divided into 3 groups (n =8 each) using a random number table:I/R group,NGF-β pretreatment group (group NGFPt) and NGF-β preconditioning group (group NGFPc).The hearts were perfused with K-H solution for 10 min (equilibration) in each group.In group I/R,the hearts were continuously perfused with K-H solution for 30 min.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min before ischemia in group NGF-Pt.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min followed by 10 min washout before ischemia in group NGFPc.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in each group.HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 10 min equilibration (baseline,T1),immediately before suspension of perfusion (T2),and at 5,30,60 and 120 min of reperfusion (T3-6).The activities of creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) in coronary effluent were measured at T1 and T3-6.Myocardial specimens were obtained at T6 for detection of myocardial apoptosis (by TUNEL) and for microscopic examination.Apoptotic index (AI) was calculated.Results Compared with the baseline value at T1,+ dp/dtmax was significantly decreased,and LVEDP and activities of CK-MB and LDH were increased at T3-6 in each group,LVDP and HR were decreased at T3-6 in group I/R,LVDP was decreased at T3,4 in group NGFPt and at T3-6 in group NGFPc,and HR was increased at T2-6 in NGFPt and NGF Pc groups (P ＜ 0.05).Compared with group I/R,LVDP,+ dp/dtmax and HR were significantly increased and LVEDP and activities of CK-MB and LDH and AI were decreased in NGFPt and NGFPc groups (P ＜ 0.05).Compared with group NGFPc,LVDP,+ dp/dtmax and HR were significantly increased,while the LDH activity and AI were decreased (P ＜ 0.05) and no significant changes were found in LVEDP and CK-MB activity in group NGFPt (P ＞ 0.05).The pathologic changes of myocardium were significantly reduced in NGFPt and NGFPc groups as compared with I/R group.Conclusion Pretreatment with 0.1 μg/ml NGF-β attenuates I/R injury in isloated rat hearts,and the efficacy is superior to that of NGF-β preconditioning.

Objective To evaluate the effects of nerve growth factor-beta (NGF-β) pretreatment on cell apoptosis induced by endoplasmic reticulum stress during ischemia-reperfusion in isolated rat hearts.Methods Male Sprague-Dawley rats,weighing 180-220 g,were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.The hearts were excised and perfused in a Langendorff apparatus with K-H solution aerated with 95％ O2 and 5％ CO2 at 37 ℃.Thirty-two isolated rat hearts were randomly divided into 4 groups (n =8 each) using a random number table:control group (group C),I/R group,NGF-β pretreatment group (group N) and NGF-β combined with K252a (trkA receptor antagonist) pretreatment group (group N + K).In group C,the hearts were continuously perfused with K-H solution for 195 min.The hearts were perfused with K-H solution for 45 min in group I/R.In N and N + K groups,the hearts were perfused with K-H solution for 15 min,and then with K-H solution containing 0.1 μg/ml NGF-β and 0.1 μg/ml NGF-β mixed with 100 nmol/L K252a,respectively,for 30 min.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in I/R,N and N + K groups.HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 15 min equilibration (baseline) and at 5,30,60 and 120 min of reperfusion.Myocardial specimens were obtained at 120 min of reperfusion for detection of myocardial apoptosis (by TUNEL) and expression of glucose-related protein 78 (GRP78),CCAAT/enhancer-binding protein homologous protein (CHOP),and caspase-12 (by Western blot analysis).Apoptosis index was calculated.Results Compared with group C,HR,LVDP and + dp/dtmax were significantly decreased,and LVEDP,apoptosis index and expression of GRP78 and CHOP were increased in I/R and N groups,and the expression of caspase-12 was upregulated in I/R group.Compared with group I/R,HR,LVDP,and + dp/dtmax were significantly increased,and LVEDP,apoptosis index and expression of GRP78,CHOP and caspase-12 were decreased in group N,and the expression of GRP78 was down-regulated in group N + K.There was no significant difference in cardiac function indexes between group I/R and N + K.Compared with group N,HR,LVDP and + dp/dtmax were significantly decreased,and LVEDP,apoptosis index,and expression of GRP78,CHOP and caspase-12 were increased in group N + K.Conclusion NGF-β pretreatment can protect the isolated rat hearts against ischemia-reperfusion injury,and inhibition of the endoplasmic reticulum stress-triggered cell apoptosis after activating trkA receptors is involved in the mechanism.

Objective To evaluate the role of 1-phosphatidylinositol 3-kinase (PI3K) signaling pathway in nerve growth factor-beta (NGF-β)-produced mitigation of cell apoptosis induced by endoplasmic reticulum stress during hypoxia-reoxygenation (H/R) in rat cardiomyocytes.Methods H9c2 cells were seeded in 96-well plates at a density of 5× 105 cells/ml (100 μl/well).The wells were randomly divided into 5 groups (n =15 each) using a random number table: control group (group C);group H/R;NGF-β group (group N);NGF-β+NGF receptor trkA antagonist K252a group (group N+K);NGF-β+ PI3K inhibitor LY294002 group (group N+L).The cells were exposed to 95％ N2-5％ CO2 for 4 h in an anaerobic incubator, followed by reoxygenation in a standard incubator for 4 h in H/R, N, N+K and N+L groups.In addition, the cells in N, N + K and N + L groups were incubated in a standard incubator containing NGF-β, the mixture of NGF-β and K252a and the mixture of NGF-β and LY294002, respectively, during reoxygenation, and the final concentrations of NGF-β , K252a and LY294002 were 50 ng/ml, 100 nmol/L and 50 μmol/L, respectively.The cell viability was detected by using CCK-8 assay, and the cell survival rate was calculated.The cell apoptosis was examined by flow cytometry, and apoptosis rate was calculated.The expression of glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), caspase-12, Akt and phosphorylated Akt (p-Akt) was detected by Western blot.The ratio of p-Akt to Akt was calculated.Results Compared with group C, the cell survival rate was significantly decreased, and the apoptosis rate was increased in H/R and N groups, and the expression of GRP78, CHOP and caspase-12 was significantly up-regulated in group H/R, and p-Akt/Akt was significantly increased in group N (P＜0.05).Compared with group H/R, the cell survival rate was significantly increased, the apoptosis rate was decreased, the expression of GRP78, CHOP and caspase-12 was up-regulated, and p-Akt/Akt was increased in group N (P＜0.05), and no significant change was found in the parameters mentioned above in N+K and N+L groups (P＞0.05).Compared with group N, the cell survival rate was significantly decreased, the apoptosis rate was increased, the expression of GRP78, CHOP and caspase-12 was up-regulated, and p-Akt/Akt was decreased in N+K and N+L groups (P＜0.05).Conclusion NGF-β can mitigate the cell apoptosis induced by endoplasmic reticulum stress during H/R, and activation of PI3K signaling pathway is involved in the mechanism.

Objective To evaluate the effect of alveolar recruitment maneuver on the perioperative pulmonary function in the morbidly obese patients undergoing laparoscopic sleeve gastrectomy.Methods Forty morbidly obese patients of both sexes,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 18-64 yr,with body mass index ≥ 40 kg/m2,scheduled for elective laparoscopic sleeve gastrectomy,were randomly divided into either control group (group C) or alveolar recruitment maneuver group (group R) using a random number table,with 20 patients in each group.Patients in group C were treated with volume-or pressure-controlled ventilation after creation of pneumoperitoneum,maintaining the peak inspiratory pressure (Ppeak) ≤ 30 cmH2O and partial pressure of end-tidal CO2 35-40 mmHg.Patients in group R received alveolar recruitment maneuver once every 30 min starting from creation of pneumoperitoneum until the end of surgery.Patients were transfered to post-anesthesia care unit (PACU) with endotracheal tube which was extubated when the unified extubation standard was achieved in PACU.The patients who stayed in PACU for 2 h showing no indications for extubation were transfered to intensive care unit for continuous ventilation support.Immediately after intubation,immediately after creation of pneumoperitoneum,at 30,60 and 90 min of pneumoperitoneum,and at the end of pneumoperitoneum,blood samples were collected from the radial artery for blood gas analysis.Immediately after intubation,immediately after creation of pneumoperitoneum,at 30,60 and 90 min of pneumoperitoneum,at the end of surgery,and immediately before discharge from PACU,Ppeak,plateau pressure (Peat),and dynamic lung compliance were recorded.The time for achieving extubation standard and time for achieving the standard for discharge from PACU were recorded.Patients were followed up until discharge,and the feeding time and duration of hospital stay were recorded.Results Compared with group C,PaO2 and oxygenation index were significantly increased at 90 min of pneumoperitoneum,at the end of surgery,and immediately before discharge from PACU,Ppeak was decreased at 60 and 90 min of pneumoperitoneum and immediately after the end of pneumoperitoneum,Pplat was decreased at 60 and 90 min of pneumoperitoneum,the dynamic lung compliance was increased at 30,60 and 90 min of pneumoperitoneum and immediately after the end of pneumoperitoneum,and the time for achieving extubation standard,time for achieving the standard for discharge from PACU,feeding time,and duration of hospital stay were shortened in group R (P＜0.05 or 0.01).In group C,one patient did not present with indications for extubation and were transfered to intensive care unit for continuous ventilation support.Conclusion Intraoperative alveolar recruitment maneuver can effectively improve the intraoperative pulmonary function and promote the recovery of postoperative pulmonary function in the morbidly obese patients undergoing laparoscopic sleeve gastrectomy.

ObjeCtive To investigate the value of protein biochip in the diagnosis of Lyme borreliosis. Methods The serum IgM and IgG antibodies against flagellin were detected by flagellin coated-and N-hydroxysuccinimide (NHS) modified-biochips in 82 patients with neuroborreliosis (NB),35 patients with erythema migrans (EM) and 44 normal controls.The results were compared with those from enzyme-linked immunosorbent assays (ELISA) and Western blot.Results The levels of both IgM and IgG antibodies were significantly higher in the NB patients than those in the normal controls (P