Effect of long-chain fat emulsion on perioperative nutritional status of patients with low rectal cancer / 肿瘤研究与临床

Objective:To explore the effect of long-chain fat emulsion in parenteral nutrition therapy on the perioperative nutritional status of patients with low rectal cancer.Methods:A total of 204 patients who underwent rectal cancer surgery in the Third Hospital of Shanxi Medical University from January 2017 to June 2020 were retrospectively analyzed. The patients were divided into two groups according to the specific nutritional treatment methods, 100 cases in the study group used long-chain fat emulsion for parenteral nutrition support, and 104 cases in the control group used medium- and long-chain fat emulsion injection. After admission, the nutritional status of patients were evaluated according to the results of Scored Patient-Generated Subjective Global Assessment (PG-SGA) and related laboratory tests. At 7th day before the operation, the patients were treated with nutrition and electrolyte support. Parenteral nutrition and enteral nutrition combined treatment and early enteral nutrition were given after the operation. The albumin, prealbumin, retinol-binding protein, total cholesterol and body mass index (BMI) at 7th day before the operation, 1st day after the operation and 7th day after the operation and the patient's first exhaust time after surgery, occurrence of postoperative complications, postoperative fever and total hospital stay were recorded and compared between the two groups.Results:Postoperative first exhaust time [(42±11) h vs. (54±10) h], fever time [(48±8) h vs. (57±7) h], total hospital stay [(16.0±0.7) d vs. (18.0±0.9) d)], resting energy expenditure at the 7th day after surgery [(5 326±589) kJ/d vs. (5 840±599) kJ/d] and total cholesterol at the 7th day after surgery [(4.8±0.3) mmol/L vs. (5.0± 0.4) mmol/L] in the study group were lower than those in the control group, and albumin [(33±3) g/L vs. (28± 3) g/L], prealbumin [(0.189±0.041) g/L vs. (0.164±0.037) g/L] and retinol-binding protein [(0.039±0.016) g/L vs. (0.032±0.013) g/L] at the 7th day after surgery in the study group were higher than those in the control group, and the differences between the two groups were statistically significant (all P < 0.05). There was no statistical difference in other detection indexes between the two groups (all P > 0.05). Conclusion:The use of long-chain fat emulsion in low rectal cancer patients with malnutrition during the perioperative period may be more conducive to the recovery of the body.

Effects of structured fat emulsion and medium / long chain fat emulsion on patients after hepatectomy / 中华肝胆外科杂志

Objective To analyze the effects of structured fat emulsion and medium/long chain fat emulsion on blood lipids,immune cells and acute inflammation after hepatectomy for hepatocellular carcinoma.Methods Total of 60 patients with hepatocellular carcinoma who underwent hepatectomy in Henan People's Hospital (Zhengzhou University People's Hospital) from January 2013 to March 2017 were divided into experimental group (using structured fat emulsion) and control group (using medium/long chain fat emulsion),30 cases in each group.Triglyceride (TG),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C),total cholesterol (TC),T lymphocyte level,fibrinogen (FIB),C-reactive protein (CRP),prostaglandin E2 (PGE2) were detected before and 2,4,6 and 8 days after infusion.Results There were no significant differences in LDL-C,HDL-C,TG and TC between the two groups before infusion (P＞0.05).On the 2nd day of parenteral nutrition infusion,the level of blood lipids in both groups was higher than experimental group before infusion;on the 4th,6th and 8th day of infusion,LDL-C,HDL-C,TG and TC in the control group were higher than those in the experimental group (P＜0.05).After parenteral nutrition infusion,the levels of CD3+,CD4+,CD8+,CD4+/CD8+ in both groups were higher than experimental group before infusion,and the experimental group was higher than the control group,the differences were statistically significant (P＜0.05).Compared with before infusion,level of FIB,CRP and PGE2 began to increase on the 2nd day of infusion,and the differences were statistically significant (P＜0.05).On the 2nd,4th,6th and 8th day,CRP in the control group was higher than experimental group.And resepeatively (19.12±5.84) mg/ml vs.(13.76±2.36) mg/ml,(31.67±8.68) mg/ml vs.(17.21±2.66) mg/ml,(22.15±8.33) mg/ml vs.(12.48±0.63) mg/ml,(9.65±4.66) mg/ml vs.(7.52±0.99) mg/ml,and PGE2 were also higher than that in the experimental group (P＜0.05).Conclusion Structured fat emulsion is superior to medium/long chain fat emulsion in improving blood lipid,immune cells and inflammatory reaction in patients after hepatectomy.

Intravenous lipid emulsion therapy for cardiac arrest and refractory ventricular tachycardia due to multiple herb intoxication

Herbal products have been used for therapeutic purposes for a long time. However, many herbs can be toxic and even life-threatening. If refractory ventricular tachycardia (VT) is caused by herbal products and shows no response to conventional therapy, intravenous lipid emulsion (ILE) therapy can be considered. We report a case of herbal intoxication leading to refractory VT, which was successfully treated with ILE therapy. A 36-year-old woman with aplastic anemia presented with mental changes. She had taken an unknown herbal decoction three days before visiting the hospital. Soon after coming to the hospital, she went into cardiac arrest. Cardiopulmonary resuscitation was performed, and return of spontaneous circulation with VT was achieved. Synchronized cardioversion was then performed and amiodarone was administered. However, VT with pulse continued, so ILE therapy was attempted, which led to the resolution of VT.

Role of mitochondrion-dependent apoptosis in reduction of bupivacaine-induced cardiotoxicity by lipid emulsion in rats / 中华麻醉学杂志

Objective@#To evaluate the role of mitochondrion-dependent apoptosis in reduction of bupivacaine-induced cardiotoxicity by lipid emulsion in rats.@*Methods@#Forty-five healthy adult male Sprague-Dawley rats, weighing 300-350 g, were divided into 3 groups by a random number table method: sham operation group (Sham group, n=5), bupivacaine group (B group, n=20), and lipid emulsion group (L group, n=20). Cardiac arrest was induced by intravenously injecting 0.4% bupivacaine 30 mg/kg over 20 s to establish the cardiotoxicity model.Twenty percent lipid emulsion was intravenously injected in a dose of 5 ml/kg during resuscitation in group L, and normal saline was intravenously injected in a loading dose of 5 ml/kg during resuscitation in group B, followed by a 3-min infusion of 1 ml·kg-1·min-1in two groups.The successful resuscitation and survival rate at 120 min of return of spontaneous circulation (ROSC) were recorded.Systolic blood pressure, heart rate, mean arterial pressure, rate-pressure product (RPP) and ratio of RPP at each time point after recovery of spontaneous heart beat to baseline value (RPPh) were recorded every 10 min after ROSC.The time from administration to cardiac arrest (T0), time from beginning of cardiopulmonary resuscitation to appearance of the first spontaneous heart beat (Ts) and time from beginning of cardiopulmonary resuscitation to appearance of ROSC (Tr) were recorded.Rats were sacrificed at 120 min of ROSC, and left ventricular tissues were obtained for determination of the expression of Bax, Bcl-2, cleaved caspase-9, cleaved caspase-3, cytochrome C (Cyt c) in cytoplasm and mitochondria (by Western blot) and expression of Bax and Bcl-2 mRNA (by real-time polymerase chain reaction) and for examination of myocardial ultrastructure.@*Results@#Compared with Sham group, the expression of Bcl-2 protein and mRNA and mitochondrial Cyt c was significantly down-regulated, and the expression of Bax protein and mRNA, cleaved caspase-9, cleaved caspase-3 and cytoplasmic Cyt c was up-regulated in B group (P<0.05). Compared with B group, the rate of successful resuscitation and survival rate were significantly increased, Tr was shortened, systolic blood pressure, heart rate, RPP and RPPh were increased after ROSC, the expression of Bcl-2 protein and mRNA and mitochondrial Cyt c was up-regulated, the expression of Bax protein and mRNA, cleaved caspase-9, cleaved caspase-3 and cytoplasmic Cyt c was down-regulated (P<0.05), no significant change was found in To or Ts (P>0.05), and the pathological changes of myocardium were significantly attenuated in L group.@*Conclusion@#The mechanism by which lipid emulsion reduces bupivacaine-induced cardiotoxicity may be related to inhibiting mitochondrion-dependent apoptosis in rats.

Role of mitochondrion-dependent apoptosis in reduction of bupivacaine-induced cardiotoxicity by lipid emulsion in rats / 中华麻醉学杂志

Objective To evaluate the role of mitochondrion-dependent apoptosis in reduction of bupivacaine-induced cardiotoxicity by lipid emulsion in rats.Methods Forty-five healthy adult male Sprague-Dawley rats,weighing 300-350 g,were divided into 3 groups by a random number table method:sham operation group (Sham group,n =5),bupivacaine group (B group,n =20),and lipid emulsion group (L group,n =20).Cardiac arrest was induced by intravenously injecting 0.4％ bupivacaine 30mg/kg over 20 s to establish the cardiotoxicity model.Twenty percent lipid emulsion was intravenously injected in a dose of 5 ml/kg during resuscitation in group L,and normal saline was intravenously injected in a loading dose of 5 ml/kg during resuscitation in group B,followed by a 3-min infusion of 1 ml · kg-1 · min-1 in two groups.The successful resuscitation and survival rate at 120 min of return of spontaneous circulation (ROSC) were recorded.Systolic blood pressure,heart rate,mean arterial pressure,rate-pressure product (RPP) and ratio of RPP at each time point after recovery of spontaneous heart beat to baseline value (RPPh) were recorded every 10 min after ROSC.The time from administration to cardiac arrest (T0),time from beginning of cardiopulmonary resuscitation to appearance of the first spontaneous heart beat (Ts) and time from beginning of cardiopulmonary resuscitation to appearance of ROSC (Tr) were recorded.Rats were sacrificed at 120 min of ROSC,and left ventricular tissues were obtained for determination of the expression of Bax,Bcl-2,cleaved caspase-9,cleaved caspase-3,cytochrome C (Cyt c) in cytoplasm and mitochondria (by Western blot) and expression of Bax and Bcl-2 mRNA (by real-time polymerase chain reaction) and for examination of myocardial ultrastructure.Results Compared with Sham group,the expression of Bcl-2 protein and mRNA and mitochondrial Cyt c was significantly down-regulated,and the expression of Bax protein and mRNA,cleaved caspase-9,cleaved caspase-3 and cytoplasmic Cyt c was up-regulated in B group (P＜0.05).Compared with B group,the rate of successful resuscitation and survival rate were signif-icantly increased,Tr was shortened,systolic blood pressure,heart rate,RPP and RPPh were increased after ROSC,the expression of Bcl-2 protein and mRNA and mitochondrial Cyt c was up-regulated,the expression of Bax protein and mRNA,cleaved caspase-9,cleaved caspase-3 and cytoplasmic Cyt c was downregulated (P＜0.05),no significant change was found in To or Ts (P＞0.05),and the pathological changes of myocardium were significantly attenuated in L group.Conclusion The mechanism by which lipid emulsion reduces bupivacaine-induced cardiotoxicity may be related to inhibiting mitochondrion-dependent apoptosis in rats.

Role of GSK-3β in lipid emulsion-induced inhibition of bupivacaine-induced apoptosis in cardiomyocytes of rats: evaluation using RNA interference adenovirus infection method / 中华麻醉学杂志

Objective To evaluate the role of glycogen synthase kinase 3 beta (GSK-3β) in lipid emulsion-induced inhibition of bupivacaine-induced apoptosis in cardiomyocytes of rats using RNA interference (RNAi) adenovirus infection method.Methods H9C2 cells were transferred into 96-well cell plates at a density of 1× 105 cells/ml after culture and then divided into 8 groups (n =10 each) using a random number table:control group (group C),bupivacaine group (group B),lipid emulsion group (group LE),bupivacaine plus lipid emulsion group (group B+LE),control plus GSK-3βRNAi adenovirus (GSK-3βi) group (group C+GSK-3βi),bupivacaine plus GSK-3βi group (group B+GSK-3βi),lipid emulsion plus GSK-3βi group (group LE+GSK-3βi) and bupivacaine plus lipid emulsion plus GSK-3βi group (group B+LE+GSK-3βi).ln B,LE and B+LE groups,the cells were incubated with culture medium containing 1 mmol/L bupivacaine,1％ lipid emulsion and 1 mmol/L bupivacaine plus 1％ lipid emulsion,respectively.In C+GSK-3βi,B+GSK-3βi,LE+GSK-3βi and B+LE+GSK-3βi groups,the cells were incubated with the drugs mentioned above on 2nd day after being infected by adenovirus.At 24 h after incubation with drugs,the expression of Bax and Bcl-2 was determined by Western blot,and the apoptosis rate was calculated using DAPI staining.Results Compared with group C,the expression of Bax was significantly upregulated,the expression of Bcl-2 was down-regulated,and the apoptosis rate was increased in group B (P＜0.05).Compared with group B,the expression of Bax was significantly down-regulated,the expression of Bcl-2 was up-regulated,and the apoptosis rate was decreased in group B+LE (P＜0.05).Compared with group B+LE,the expression of Bax was significantly up-regulated,the expression of Bcl-2 was downregulated and the apoptosis rate was increased in group B+LE+GSK-3βi (P＜0.05).Conclusion The mechanism by which lipid emulsion inhibits bupivacaine-induced apoptosis in cardiomyocytes of rats is associated with GSK-33.

Effect of emulsified isofluraue postconditioning on mitophagy during myocardial ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the effect of emulsified isoflurane postconditioning on mitophagy during myocardial ischemia-reperfusion (I/R) in rats.Methods Forty-eight pathogen-free healthy male Sprague-Dawley rats,aged 4-5 months,weighing 250-300 g,were divided into 4 groups (n=12 each) using a random number table:sham operation group (group S),group I/R,fat emulsion group (group F) and emulsified isoflurane postconditioning group (group EIP).Myocardial I/R was induced by occlusion of the anterior descending branch of the left coronary artery for 30 min followed by 120 min of reperfusion in pentobarbital sodium-anesthetized rats.Starting from 3 min before reperfusion,8％ emulsified isoflurane 2 ml/kg was intravenously infused over 8 min in group EIP,while 30％ fat emulsion 2 ml/kg was intravenously infused over 8 min in group F.Rats were sacrificed at the end of reperfusion,and hearts were removed for measurement of the myocardial infarct size (by 2,3,5-triphenyltetrazolium chloride staining),cell apoptosis (by TUNEL),mitochondrial membrane potential and expression of microtubule-associated protein 1 light chain 3 (LC3),Beclinl,P62,PINK1 and Parkin in cardiomyocytes (by using Western blot).Apoptosis index (AI) was calculated.Results Compared with group S,the myocardial infarct size and AI were significantly increased,the mitochondrial membrane potential was decreased,the expression of LC3,Beclinl,PINK1 and Parkin was up-regulated,and the expression of P62 was down-regulated in I/R,F and EIP groups (P＜0.05).Compared with group I/R,the myocardial infarct size and AI were significantly decreased,the mitochondrial membrane potential was increased,the expression of LC3,Beclinl,PINK1 and Parkin was down-regulated,and the expression of P62 was up-regulated in group EIP (P＜0.05).Compared with group F,the myocardial infarct size and AI were significantly decreased,the mitochondrial membrane potential was increased,the expression of LC3,Beclin1,PINK1 and Parkin was down-regulated,and the expression of P62 was up-regulated in group EIP (P＜0.05).Conclusion The mechanisin by which emulsified isoflurane postconditioning reduces myocardial I/R injury is related to inhibition of mitophagy in rats.

Effect of emulsified isoflurane post-conditioning on mitochondrial function during lung ischemia-reperfusion in rats: an in vitro experiment / 中华麻醉学杂志

Objective To evaluate the effect of emulsified isoflurane post-conditioning on the mitochondrial function during lung ischemia-reperfusion (I/R) in rats in an in vitro experiment.Methods Twenty-four SPF healthy male Sprague-Dawley rats,weighing 250-300 g,were used in the study.After the animals were anesthetized,the lungs were removed,connected to the perfusion system and then divided into 4 groups (n=6 each) using a random number table:control group (group C),group I/R,emulsified isoflurane post-conditioning group (group EI) and intralipid post-conditioning group (group IL).After 20 min of equilibration,the lungs were continuously perfused for 105 min in group C,and the lungs were subjected to 45 min ischemia followed by 60 min reperfusion to establish the model of lung I/R injury in the other three groups.During the reperfusion period,the common perfusate was used in group I/R,the perfusate containing 1.68 mmol/L emulsified isoflurane was used in group EI,and the equal volume of perfusate containing 30％ intralipid was used in group IL.At the end of the equilibration (T0),immediately after beginning of reperfusion (T1) and at 30 and 60 min of reperfusion (T2.3),the arterial oxygen partial pressure (PaO2),airway resistance,pulmonary compliance and tidal volume (VT) were recorded.The right upper lobe of the lung was removed at T3 for determination of wet to dry weight ratio (W/D ratio).The right middle lobe of the lung was removed at T3 for pathologic examination with light microscope.The contents of reactive oxygen species (ROS),NAD+ and ATP in lung tissues were detected.Results Compared with group C,the PaO2,pulmonary compliance and Vr were significantly decreased,and the airway resistance was increased at T1-3,and the W/D ratio and ROS content were increased,and NAD+ and ATP contents were decreased at T3 in I/R,EI and IL groups (P＜0.05).Compared with I/R and IL groups,the PaO2,pulmonary compliance and VT were significantly increased,and the airway resistance was decreased at T2.3,and the W/D ratio and ROS content were decreased,and NAD+ and ATP contents were increased at T3 in group EI (P＜0.05).The pathologic changes of lungs were significantly attenuated in group EI as compared with group I/R.Conclusion The mechanism by which emulsified isoflurane post-conditioning attenuates lung I/R injury is related to decrease in mitochondrial dysfunction in rats in an in vitro experiment.

A bioequivalence evaluation of long-chain triacylglycerol/medium-chain triacylglycerol emulsion in beagle dogs / 中华临床营养杂志

Objective Long-chain triacylglycerol (LCT) by three producers,each mixed with the same medium-chain triacylglycerol (MCT),were compared with the brand MCT/LCT in causing focal necrosis of hepatocytes in beagle dogs (a bioequivalence evaluation).Methods 21 beagle dogs (male,0.7-1.5 years old,10-15 kg) were used in this study.According to the sources of the LCT,the animals were divided into Group A (LCT made in China),Group B (LCT made in Japan),Group C (LCT made in Germany),and the control group (the brand 10％ MCT/LCT).Central venous port was placed via the lumber vein of the animals under general anesthesia.After 2 weeks of rehabilitation,MCT/LCT was administered through this port for 28 days at 9 g/ (kg · d) [while the routine dose used clinically was 1 g/ (kg · d)].The laboratory indexes and the pathomorphism of the liver and kidney were studied single blindly.Results Laboratory tests,including liver and kidney function,blood coagulation function and lipid metabolism,did not identify differences among emulsions with different sources of LCT.Liver biopsy at day 28 showed no focal necrosis in Group C and the control group;there was minor damage in Group B;and Group A had obvious liver necrosis.and the pathological findings of other organs are similar.No significant difference was observed in biopsies of other organs.Conclusions Emulsions with different sources of LCT varied in their damage to the liver.Generics with LCT of higher quality were equivalent to the brand MCT/LCT in terms of safety.

Role of ROS in emulsified isoflurane postconditioning-induced promotion of Nrf2∕ARE signaling pathway activation in rats with myocardial injury / 中华麻醉学杂志

Objective To investigate the role of reactive oxygen species ( ROS ) in emulsified isoflurane postconditioning?induced promotion of nuclear factor?E2 related factor 2 ( Nrf2 )∕antioxidant re?sponse element ( ARE) signaling pathway activation in rats with myocardial injury. Methods Healthy male Sprague?Dawley rats, aged 16-20 weeks, weighing 250-300 g, were heparinized and anesthetized. Their hearts were excised and perfused in a Langendorff apparatus with K?H solution. Eighty isolated rat hearts were divided into 5 groups ( n=16 each) using a random number table: control group ( group C);ischemia?reperfusion (I∕R) group; emulsified isoflurane postconditioning group (group EIP); fat emulsion postconditioning group (group FEP); N?(2?mercaptopropionyl)?glycine (a ROS scavenger) plus emulsi?fied isoflurane postconditioning group ( group N+EIP ) . Group C was perfused with K?H solution for 100 min, and the other 4 groups were subjected to 40 min of ischemia followed by 60 min of reperfusion. EIP and FEP groups were perfused for 2 min with K?H solution containing 1?68 mmol∕L emulsified isoflurane and 712 mg∕L intralipid, respectively, starting from the onset of reperfusion, and then continuously per?fused with K?H solution for 58 min. Group N+EIP was perfused for 3 min with K?H solution containing N?(2?mercaptopropionyl)?glycine 2 mmol∕L, and then emulsified isoflurane postconditioning was performed. Heart rate ( HR ) , left ventricular developed pressure ( LVDP ) , left ventricular end?diastolic pressure (LVEDP) and the maximum rate of increase of left ventricular pressure (+dp∕dtmax) were recorded at the end of equilibration and of reperfusion. At 5 min of reperfusion and the end of reperfusion, myocardial specimens were obtained for determination of ROS content. At the end of reperfusion, myocardial specimens were obtained for examination of the ultrastructure of myocardial cells and for determination of the expression of Nrf2, heme oxygenase?1 ( HO?1 ) , quinone oxidoreductase 1 ( NQO1 ) and superoxide dismutase?1 ( SOD?1) protein and mRNA in myocardial tissues. Mitochondrial injury scores ( Flameng scores) were e?valuated. Results Compared with group C, HR, +dp∕dtmax and LVDP were significantly decreased, LV?EDP , mitochondrial Flameng scores and ROS contents were increased, and the expression of Nrf2, HO?1, NQO1 and SOD?1 protein and mRNA was down?regulated at the end of reperfusion in I∕R group ( P0?05) . Compared with group EIP, HR, +dp∕dtmax and LVDP were significantly decreased, LVEDP and mitochondrial Flameng scores were increased, ROS contents at 5 min of reperfusion were decreased, ROS contents at the end of reperfu?sion were increased, and the expression of Nrf2, HO?1, NQO1 and SOD?1 protein and mRNA was down?regulated in group N+EIP ( P<0?05) . The degree of myocardial injury was reduced in group EIP as com?pared with group I∕R. There was no significant difference in the degree of myocardial injury between group N+EIP and group I∕R. Conclusion The mechanism by which emulsified isoflurane postconditioning pro?motes Nrf2?ARE signaling pathway activation is totally related to ROS in rats with myocardial injury.

Effects of emulsified isoflurane on apoptosis in human neuroblastoma cells : the role of c-Jun N-terminal kinase / 中华麻醉学杂志

Objective To investigate the effects of emulsified isoflurane on apoptosis in human neuroblastoma cells and the role of c-Jun N-terminal kinase (JNK) in it.Methods The human neuroblastoma SHSY-5Y cells were seeded in 96-well plates or dishes and then randomly divided into 8 groups using a random number table:control group(group C,n=24),different concentrations of lipid emulsion groups(LE1 groups [n =24],LE2 group [n =24] and LE3 group [n =72]),different concentrations of emulsified isoflurane groups (EI1 group [n=24],El2 group [n=24] and EI3 group [n=72]),and emulsified isoflurane + JNK inhibitor SP600125 group (group EI-SP,n =24).At 24 h after the cells were plated,in LE1-3 groups,30％ lipid emulsion was added to the culture medium with the final concentrations of 0.395 6,0.791 2 and 1.582 4 μl/ml,respectively;in EI1-3 groups,8％ emulsified isoflurane was added with the final concentrations of 0.56,1.12 and 2.24 mmol/L,respectively;in group EI-SP,emulsified isoflurane was added with the final concentration of 1.12 mmol/L,and SP600125 was added at 1 h before addition of emulsified isoflurane with the final concentration of 10 μmol/L;the cells were cultured normally in group C.At 6,12 and 24 h of incubation in EI3 and LE3 groups,and at 24 h of incubation or culture in the other groups,the morphology of cells was detected,the cell viability was measured using methyl thiazolyl tetrazolium assay,and the expression of JNK,phosphorylated JNK (p-JNK) and cytochrome c (Cyt c) was detected by Western blot.Results Compared with group C,the cell viability was significantly decreased,and the expression of p-JNK and Cyt c was significantly up-regulated at 24 h of incubation in group EI2 and at 12 and 24 h of incubation in group EI3,the cell viability was significantly decreased (P＜0.05),and no significant change was found in the expression of p-JNK and Cyt c in group EI-SP,and no significant change was found in the cell viability and expression of p-JNK and Cyt c in LE1-3 and EI1 groups (P＞0.05).Compared with group EI1,the cell viability was significantly decreased,and the expression of p-JNK and Cyt c was significantly up-regulated at 24 h of incubation in EI2.3 groups (P＜0.05).Compared with group EI2,the cell viability was significantly decreased,and the expression of p-JNK and Cyt c was significantly up-regulated at 24 h of incubation in group EI3,and the cell viability was significantly increased,and the expression of p-JNK and Cyt c was significantly down-regulated in group EI-SP (P＜0.05).There was no significant difference in JNK expression between the eight groups (P＞0.05).Conclusion High concentrations of emulsified isoflurane can induce apoptosis in neurons only when applied for a long time,while low concentrations do not have the effect when applied for a short time.The mechanism by which emulsified isoflurane induces neuronal apoptosis is related to activation of JNK pathway.

Hemodynamic changes with two lipid emulsions for treatment of bupivacaine toxicity in swines

PURPOSE: To compare the hemodynamic changes following two different lipid emulsion therapies after bupivacaine intoxication in swines. METHODS: Large White pigs were anesthetized with thiopental, tracheal intubation performed and mechanical ventilation instituted. Hemodynamic variables were recorded with invasive pressure monitoring and pulmonary artery catheterization (Swan-Ganz catheter). After a 30-minute resting period, 5 mg.kg-1 of bupivacaine by intravenous injection was administered and new hemodynamic measures were performed 1 minute later; the animals were than randomly divided into three groups and received 4 ml.kg-1 of one of the two different lipid emulsion with standard long-chaim triglyceride, or mixture of long and medium-chain triglyceride, or saline solution. Hemodynamic changes were then re-evaluated at 5, 10, 15, 20 and 30 minutes. RESULTS: Bupivacaine intoxication caused fall in arterial blood pressure, cardiac index, ventricular systolic work index mainly and no important changes in vascular resistances. Both emulsion improved arterial blood pressure mainly increasing vascular resistance since the cardiac index had no significant improvement. On the systemic circulation the hemodynamic results were similar with both lipid emulsions. CONCLUSION: Both lipid emulsions were efficient and similar options to reverse hypotension in cases of bupivacaine toxicity. .

Effect of lipid emulsion on mitochondrial energy metabolism during bupivacaine-induced myocardiotoxicity in rats / 中华麻醉学杂志

Objective To investigate the effect of lipid emulsion on mitochondrial energy metabolism during bupivacaine-induced myocardiotoxicity in rats.Methods H9c2 cells were inoculated in 96-well plates at a density of 1 × 105cells/ml, and were randomly divided into 4 groups (n =18 each) using a random number table: control group (group C) , bupivacaine group (group B) , lipid emulsion group (group LE) , and bupivacaine + lipid emulsion group (group B + LE).The cells were incubated in the normal culture medium in group C.In group LE, the cells were incubated in the culture medium containing 1％ lipid emulsion.In group B, the cells were incubated in the culture medium containing 1 mmol/L bupivacaine.In group B + LE, the cells were incubated in the culture medium containing 1 mmol/L bupivacaine and 1％ lipid emulsion.After 24 h of incubation, the contents of ATP, ADP, and AMP were measured by high-performance liquid chromatography, and apoptotic rate was calculated by Hoechst33342/ PI staining.Results Compared with group C, the contents of ATP, ADP and AMP were significantly decreased, and apoptotic rate was increased in group B (P＜0.05), and the contents of ATP and ADP in group LE and ATP content in group B + LE were increased, and no significant changes were found in apoptotic rate in LE and B+LE groups (P＞0.05).Compared with group C, the contents of ATP, ADP and AMP were significantly increased, and apoptotic rate was decreased in LE and B+LE groups (P＜ 0.05).Compared with group LE, the contents of ATP, ADP and AMP were significantly decreased (P＜ 0.05), and no significant change was found in apoptotic rate in group B+LE (P＞0.05).Conclusion The mechanism by which lipid emulsion reduces bupivacaine-induced myocardiotoxicity may be associated with improved mitochondrial energy metabolism in rats.

Role of mitochondrial permeability transition pore in lipid emulsion-induced inversion of bupivacaine myocardiotoxicity in rats / 中华麻醉学杂志

Objective To evaluate the effect of mitochondrial permeability transition pore (mPTP) in lipid emulsion-induced inversion of bupivacaine myocardiotoxicity in rats.Methods H9c2 cells were inoculated in 6-well plates at a density of 105 cells/ml, and were randomly divided into 4 groups (6 wells in each group, 2 ml/well) using a random number table: control group (group C) , bupivacaine group (group B) , lipid emusion + bupivacaine group (group LB) , and lipid emusion + bupivacaine + atractyloside group (group LBA).Phosphate buffer solution 100 μl was added to the culture medium in group C.In group B, bupivacaine was added to the culture medium with the final concentration of 1 mmol/L.In group LB, lipid emusion and bupivacaine were added to the culture medium with the final concentrations of 1％ and 1 mmol/L, respectively.In group LBA, lipid emusion, bupivacaine and atractyloside (an mPTP opener) were added to the culture medium with the final concentrations of 1％, 1 mmol/L and 30 μmol/L, respectively.All the cells were incubated for 24 h.After the end of incubation, the expression of Bcl-2, Bax, phosphorylated Bad (p-Bad) , caspase-3, activated caspase-3, caspase-9,activated caspase-9 and cytochrome c (Cyt c) was detected using Western blot.The expression of Bcl-2 mRNA, Bax mRNA, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was detected using real-time reverse transcriptase polymerase chain reaction.The ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were calculated.Results Compared with group C,the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/ Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group B (P＜0.05) , and no significant change was found in the parameters mentioned above in group LB (P＞0.05).Compared with group B, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly decreased, the expression of p-Bad was up-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was down-regulated in LB and LBA groups (P＜ 0.05).Compared with group LB, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group LBA (P ＜ 0.05).Conclusion The mechanism underlying lipid emulsioninduced inversion of bupivacaine myocardiotoxicity is related to inhibited mPTP opening in rats.

Effects of emulsified isoflurane preconditioning on lung ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To investigate the effects of emulsified isoflurane preconditioning on lung ischemia-reperfusion (Ⅰ/R) injury in rats.Methods Thirty-two male Sprague-Dawley rats,weighing 250-320 g,were randomly divided into 4 groups (n =8 each) using a random number table:sham operation group (group S),group Ⅰ/R,intralipid group (group IL),and emulsified isoflurane preconditioning group (group EI).Lung Ⅰ/R was produced by clamping the left hilum of lungs for 1 h followed by 2 h of reperfusion in anesthetized rats.In group S,intraperitoneal normal saline 10.5 ml/kg was injected,but the hilum was only exposed 24 h later.In groups Ⅰ/R,IL and EI,intraperitoneal normal saline 10.5 ml/kg,30％ intralipid 10.5 ml/kg and 8％ emulsified isoflurane 10.5 ml/kg were injected intraperitoneally,respectively,and then the model was established 24 h later.At the end of reperfusion,blood samples were collected from the left ventricle for measurement of PaO2,PaCO2,plasma malondialdehyde (MDA) concentration (by ELISA).The rats were then sacrificed,and left lungs were removed immediately for microscopic examination and for determination of myleoperoxidase (MPO) activity (using colorimetric method) in pulmonary specimens.Wet/dry lung weight ratio (W/D ratio) was calculated.Results Compared with group S,W/D ratio,MPO activity,plasma MDA concentration and PaCO2 were significantly increased,and PaO2 was decreased in the other three groups.Compared with group Ⅰ/R,W/D ratio,MPO activity,plasma MDA concentration and PaCO2 were significantly decreased,and PaO2 was increased in IL and EI groups.W/D ratio,MPO activity,plasma MDA concentration and PaCO2 were significantly lower,and PaO2 was higher in group EI than in group IL.The pathological changes of the lung were significantly attenuated in group EI compared with group Ⅰ/R.Conclusion Emulsified isoflurane preconditioning can reduce lung Ⅰ/R injury in rats,and inhibition of pulmonary inflammatory responses and of production of oxygen free radicals are involved in the mechanism.

Effect of emulsified isoflurane postconditioning on Nrf2-ARE signaling pathway during myocardial ischemia-reperfusion in rats in vitro / 中华麻醉学杂志

Objective To evaluate the effect of emulsified isoflurane postconditioning on nuclear factor?E2 related factor 2 ( Nrf2 )?antioxidant response element ( ARE ) signaling pathway during myocardial ischemia?reperfusion ( I∕R ) in rats in vitro. Methods Healthy male Sprague?Dawley rats, aged 4-5 months, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1% amobarbital sodium 40 mg∕kg. Their hearts were excised and perfused in a Langendorff apparatus with K?H solution. Thirty?two isolated rat hearts were randomly divided into 4 groups ( n=8 each ) using a random number table: control group (group C), group I∕R, emulsified isoflurane postconditioning group (EIP group) and fat emulsion group ( group F) . After 20 min of equilibration, group C was continuously perfused with K?H solusion for 100 min. Group I∕R underwent 40 min of ischemia at 32 ℃, followed by reperfusion for 60 min. In EIP and F groups, after undergoing 40 min of global ischemia, the isolated hearts were perfused for 2 min with K?H solution containing 1.68 mmol∕L emulsified isoflurane and 712 mg∕L intralipid, respectively, starting from the onset of reperfusion, and then were continuously perfused with K?H solution containing oxygen at 37 ℃ for 58 min. Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end?diastolic pressure ( LVEDP ) , and positive maximal pressure of left ventricular increase (+dp∕dtmax ) were recorded at the end of equilibration and reperfusion. At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase?1 ( HO?1) , quinone oxidoreductase 1 ( NQO1) , and superoxide dismutase 1 ( SOD1) and mRNA expression using Western blot and real?time PCR. Results Compared with group C, HR, +dp∕dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I∕R and F groups, LVDP was significantly decreased, LVEDP was increased, and no significant changes were found in HR and +dp∕dtmax at the end of reperfusion in EIP group, and Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was down?regulated at the end of reperfusion in I∕R, EIP and F groups. Compared with group I∕R, HR, +dp∕dtmax and LVDP were significantly increased, and LVEDP was decreased at the end of reperfusion in EIP and F groups, Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was significantly up?regulated at the end of reperfusion in EIP group, and Nrf2, HO?1, NQO1 and SOD1 mRNA expression was significantly up?regulated, Nrf2 and HO?1 expression was up?regulated, and no significant changes were found in NQO1 and SOD1 expression at the end of reperfusion in group F. Compared with group EIP, HR, +dp∕dtmax and LVDP were significantly decreased, LVEDP was increased, and Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was down?regulated in group F. Conclusion Emulsified isoflurane postconditioning attenuates myocardial I∕R injury probably by activating Nrf2?ARE signaling pathway in isolated rat hearts.

Role of Nrf2-ARE signaling pathway in mitigation of anoxia/reoxygenation-induced damage to rat cardiomyocytes by emulsified isoflurane postconditioning / 中华麻醉学杂志

Objective To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway in mitigation of anoxia/reoxygenation (A/R)-induced damage to rat cardiomyocytes by emulsified isoflurane postconditioning.Methods Primarily cultured cardiomyocytes obtained from Sprague-Dawley rats,aged 16-20 weeks,were seeded into the laminin pre-treated 6-well plates at a density of 104/cm2 and cultured for 20 h.The cells were randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),group A/R,and emulsified isoflurane postconditioning group (group EI).The cells were cultured for 110 min without any treatment in group C.The cells were exposed to 45 min anoxia followed by 60 min reoxygenation in A/R and EI groups.After 45 min of hypoxia,the cells were cultured with emulsified isoflurane 1.68 mmol/L for 5 min before onset of reoxygenation in group EI.At the end of reoxygenation,transmission electron microscope was used to observe the ultrastructure of myocardial cells,and mitochondrial function was scored.The mRNA and protein expression of Nrf2,heme oxygenase-1 (HO-1),superoxide dismutase 1 (SOD1),and quinone oxidoreductase 1 (NQO1) was detected by real-time PCR and Western blot.Laser scanning confocal microscopy was used to detect Nrf2 nuclear translocation at the end of incubation,and Nrf2 activity was recorded in the nucleus of myocardial cells.Results Compared with group C,the mitochondrial function score was significantly increased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was down-regulated,and Nrf2 activity in the nucleus was enhanced in A/R and EI groups.Compared with group A/R,the mitochondrial function score was significantly decreased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was up-regulated,and Nrf2 activity in the nucleus was enhanced in group EI.Conclusion The mechanism by which emulsified isoflurane postconditioning mitigates A/R-induced damage to rat cardiomyocytes is related to induced nuclear translocation of Nrf2 and activated Nrf2-ARE signaling pathway.

Effect of emulsified isoflurane preconditioning on renal ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To evaluate the effect of 8% emulsified isoflurane preconditioning on renal ischemia-reperfusion (I/R) injury in rats.Methods Thirty-two male Sprague-Dawley rats,aged 10-13 weeks,weighing 220-300 g,were randomly divided into 4 groups (n =8 each):sham operation group (group S); I/R group;emulsified isoflurane preconditioning group (group E) ; intralipid preconditioning group (group I).Renal ischemia was induced by occlusion of the left renal pedicle for 45 min with atraumatic microclips followed by 3 h reperfusion.8 % emulsified isoflurane and 30 % intralipid 4 ml· kg-1· h-1 were infused intravenously for 30 min followed by 15 min washout before renal UR in groups E and I,respectively.Arterial blood samples were taken at 3 h of reperfusion to determine the concentrations of serum creatinine (Cr),cystatin C (Cys C),TNF-α,IL-6 and IL-10.The animals were then sacrificed and left kidneys were removed and stained with hematoxylin-eosin for microscopic examination and assessment of necrosis of renal proximal convoluted tubules (0 =normal,4 =necrosis of whole segment of proximal convoluted tubules).Results Compared with group S,the serum Cr,Cys C,TNF-α,IL-6 and IL-10 concentrations and severity of necrosis of renal proximal convoluted tubules were significantly increased in groups I/R,E and I (P ＜ 0.05).The serum Cr,Cys C,TNF-α and IL-6 concentrations and severity of necrosis of renal proximal convoluted tubules were significantly lower,while the serum IL-10 concentration was higher in group E than in groups I/R and I (P ＜0.05).There was no significant difference in the indexes mentioned above between groups L and I/R (P ＞ 0.05).The damage to renal tissues was less serious in group E than in groups I/R and L.Conclusion Preconditioning with 8 % emulsified isoflurane can attenuate renal I/R injury by inhibiting inflammatory responses in rats.

Effect of emulsified isoflurane anesthesia on expression of hippocampal amyloid-beta protein and phosphorylation of Tau protein in rats / 中华麻醉学杂志

Objective To investigate the effect of emulsified isoflurane anesthesia on the expression of hippocampal amyloid-beta protein (Aβ) and phosphorylation of Tau protein in rats.Methods Seventy-two healthy Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were randomly divided into 3 groups:normal control group (group C,n =12),fat emulsion group (group E,n =12),and 8％ emulsified isoflurane group (group EI,n =48).30％ fat emulsion and 8％ emulsified isoflurane 0.15 ml/100 g were slowly injected via the tail vein in groups E and EI,respectively.Morris water maze test was performed 6 days before anesthesia.Twelve animals in each group were chosen at 2 h after administration (T1) in E group,at the corresponding time points in C group,or at T1 and 1,7 and 14 days after administration (T2-4) in EI group and underwent spatial probe tests,and the escape latency was measured.The rats were anesthetized with intraperitoneal 1％ pentobarbital sodium 4 g/100 g after the end of Morris water maze test.Blood samples were taken for determination of the plasma S100 protein concentration.The rats were then sacrificed and brains were removed for determination of the expression of hippocampal Aβ and phosphorylated Tau (p-Tau) protein by immunohistochemistry.Results The escape latency and swimming distance were significantly shorter on 3rd,4th and 5th days than those on 1st day of place navigation test before anethesia (P ＜ 0.01).Compared with group C,the escape latency and swimming distance were significantly prolonged and the time of staying at the original platform quadrant was shortened at T1,and the plasma S100 protein concentration was decreased at T4 in group EI (P ＜ 0.05),and no significant change was found in the each parameter of Morris water maze test,plasma S100 protein concentration,and expression of Aβ and p-Tau protein in group E (P ＞ 0.05).The escape latency and swimming distance were significantly shorter and the time of staying at the original platform quadrant was longer at T2-T4 than at T1 in group EI (P ＜ 0.05).Conclusion Emulsified isoflurane anesthesia exerts no effect on the expression of hippocampal Aβ and phosphorylation of Tau protein,indicating that hippocampal Aβ and Tau protein are not involved in emulsified isoflurane anesthesia-induced cognitive dysfunction in rats.

Terapia lipídica com dois agentes na intoxicação por ropivacaína: estudo experimental em suínos / Lipid therapy with two agents in ropivacaine-induced toxicity: experimental study in swine / Terapia lipídica con dos agentes en la intoxicación por ropivacaína: estudio experimental en cerdos

OBJETIVO: Comparar alterações hemodinâmicas após intoxicação com ropivacaína seguida de terapia com duas emulsões lipídicas em suínos. MÉTODO: Suínos da raça Large White foram anestesiados com tiopental, intubados e mantidos em ventilação mecânica. Variáveis hemodinâmicas de repouso foram registradas através de pressão invasiva e cateterização da artéria pulmonar. Após 30 minutos, 7 mg.kg-1 de ropivacaína foram injetados por via venosa e novas medidas hemodinâmicas foram feitas em um minuto; os animais foram então aleatoriamente alocados em três grupos e receberam: 4 mL.kg-1 de solução salina, 4 mL.kg-1 de solução lipídica com triglicérides de cadeia longa e 4 mL.kg-1 de solução lipídica com triglicérides de cadeia média e longa. As alterações hemodinâmicas foram reavaliadas aos cinco, 10, 15, 20 e 30 minutos. RESULTADOS: A intoxicação pela ropivacaína causou queda da pressão arterial e do índice cardíaco, principalmente, sem importantes alterações das resistências vasculares. A terapia com as emulsões lipídicas restaurou a pressão arterial através, principalmente, do aumento das resistências vasculares, uma vez que o índice cardíaco não apresentou melhoria expressiva. A emulsão lipídica com triglicérides de cadeia média causou aumento superior das resistências vasculares, sobretudo pulmonares. CONCLUSÃO: Nos grupos que receberam emulsões lipídicas os resultados hemodinâmicos foram melhores do que no grupo controle; não foram observadas diferenças da pressão arterial sistêmica e do índice cardíaco entre os animais que receberam a solução com triglicérides de cadeia longa e a mistura de triglicérides de cadeia média e longa.

BACKGROUND AND OBJECTIVE: Compare hemodynamic changes after ropivacaine-induced toxicity followed by treatment with two lipid emulsions in swine. METHODS: Large White pigs were anesthetized with thiopental, followed by intubation, and kept on mechanical ventilation. Hemodynamic variables at rest were recorded with invasive pressure monitoring and pulmonary artery catheterization. After 30 minutes, 7 mg.kg-1 ropivacaine were injected intravenously and new hemodynamic measurements were performed within one minute. The animals were then randomly allocated into three groups and received: 4 mL.kg-1 saline solution, or 4 mL.kg-1 lipid emulsion with long-chain triglycerides, or 4 mL.kg-1 lipid emulsion with long-and medium-chain triglycerides. Hemodynamic changes were reevaluated at 5, 10, 15, 20 and 30 minutes. RESULTS: Ropivacaine-induced toxicity mainly caused a drop in blood pressure and cardiac index without significant changes in vascular resistance. Therapy with lipid emulsions restored blood pressure primarily through increased vascular resistance, as cardiac index showed no significant improvement. Lipid emulsion with medium-chain triglycerides caused a greater increase in vascular resistance, particularly pulmonary. CONCLUSION: In groups receiving lipid emulsions, hemodynamic results were better than in control group. There were no differences in systemic arterial pressure and cardiac index between animals receiving lipid emulsion with long-chain triglycerides and mixed long- and medium-chain triglycerides.

JUSTIFICATIVAS Y OBJETIVO: Comparar las alteraciones hemodinámicas después de la intoxicación con ropivacaína seguida de terapia con dos emulsiones lipídicas en cerdos. MÉTODO: Cerdos de la raza Large White que fueron anestesiados con tiopental, intubados y mantenidos bajo ventilación mecánica. Las variables hemodinámicas de reposo se registraron por medio de la presión invasiva y la cateterización de la arteria pulmonar. Después de 30 minutos, se inyectaron 7 mg.kg-1 de ropivacaína por vía venosa y nuevas medidas hemodinámicas se tomaron en un minuto. Los animales se dividieron entonces aleatoriamente en tres grupos y recibieron: 4 mL.kg-1 de solución salina, 4 mL.kg-1 de solución lipídica con triglicéridos de cadena larga y 4 mL.kg-1 de solución lipídica con triglicéridos de cadena media y larga. Las alteraciones hemodinámicas fueron nuevamente calculadas a los 5, 10, 15, 20 y 30 minutos. RESULTADOS: La intoxicación por ropivacaína ha causado la caída de la presión arterial y del índice cardíaco, principalmente, sin alteraciones importantes de las resistencias vasculares. La terapia con las emulsiones lipídicas ha restaurado la presión arterial principalmente por medio del aumento de las resistencias vasculares, una vez que el índice cardíaco no tuvo una mejoría expresiva. La emulsión lipídica con triglicéridos de cadena media causó un aumento superior de las resistencias vasculares, sobre todo en las pulmonares. CONCLUSIONES: En los grupos que recibieron emulsiones lipídicas, los resultados hemodinámicos fueron mejores que en el grupo control. No se observaron diferencias de la presión arterial sistémica y del índice cardíaco entre los animales que recibieron la solución con triglicéridos de cadena larga y la mezcla de triglicéridos de cadena media y larga.