Risk factors for early acute lung injury after pediatric living-related liver transplantation and the predictive value / 中华麻醉学杂志

Objective:To identify the risk factors for early acute lung injury (ALI) after living-related liver transplantation in pediatric patients and evaluate the value of the risk factors in prediction of ALI.Methods:Perioperative data of patients were obtained through the electronic medical records system. Patients were divided into non-ALI group and ALI group according to whether ALI occurred within the first week after surgery. The risk factors of which P values were less than 0.05 would enter the multiple logistic regression analysis to stratify ALI-related risk factors. Area under the ROC curve was used to analyze the value of the risk factors in prediction of postoperative ALI. Results:A total of 67 patients were enrolled, including 45 cases in non-ALI group and 22 cases in ALI group. Increased intraoperative blood transfusion volume and up-regulated expression of miR-122 and miR-21 were independent risk factors for the occurrence of postoperative ALI ( P<0.05), and the area under the ROC curve (95% confidence interval) of serum miR-122 and miR-21 expression was 0.946 (0.875 to 1.00), with sensitivity and specificity of 95% and 90%, respectively. Conclusions:Increased intraoperative blood transfusion volume and up-regulated expression of serum miR-122 and miR-21 are independent risk factors for early postoperative ALI, and serum miR-122 and miR-21 levels have a high value in prediction of the development of postoperative ALI in pediatric patients undergoing living-related liver transplantation.

Relationship between serum exosomes and microglial pyroptosis during brain injury in a young rat model of hepatic ischemia-reperfusion / 中华麻醉学杂志

Objective:To evaluate the relationship between serum exosomes and microglial pyroptosis during brain injury in a young rat model of hepatic ischemia-reperfusion (I/R).Methods:Forty-six clean-grade healthy male Sprague-Dawley rats, aged 2-3 weeks, weighing 40-60 g, were allocated into 4 groups using a random number table method: sham operation group (group S, n=10), hepatic I/R group (group I/R, n=13), treatment with serum exosome in sham-operated young rat group (group S-Exosome, n=10), and treatment with serum exosomes in young rats with I/R group (group I/R-Exosome, n=13). The common trunk of the portal vein, left hepatic artery and bile duct was clamped for 60 min resulting in ischemia of 70% of the liver in anesthetized animals.After 6 h of reperfusion, the serum was collected to extract exosomes in S group and I/R group, and the serum exosome suspension 100 μl of S group and I/R group was injected through the tail vein in S-Exosome group and I/R-Exosome group, respectively.The expression of serum exosome marker proteins CD9 and CD81 was determined by Western blot in S group and I/R group.Serum and hippocampi were obtained from each group at 6 h after the corresponding treatment.The expression of NOD-like receptor protein 3 (NLRP3), gasdermin D (GSDMD), pro-caspase-1, cleaved-caspase-1, and apoptosis-associated speck-like protein containing CARD (ASC) in the hippocampus was detected using Western blot, and the expression of GSDMD in hippocampal tissues was determined by immunohistochemistry.The levels of interleukin-1beta (IL-1β), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) in serum and hippocampal tissues and S100β and NSE in serum were determined by enzyme-linked immunosorbent assay.In group I/R-Exosome, 3 rats were selected, and their serum exosomes were extracted, labeled with PKH26 red fluorescence and then injected via the tail vein, and the co-localization between exosomes and microglia was identified by immunofluorescence technique. Results:Compared with group S, the expression of serum CD9 and CD81 was significantly up-regulated in group I/R, the expression of NLRP3, GSDMD, cleaved-caspase-1 and ASC in hippocampal tissues was significantly up-regulated, the levels of IL-18, IL-1β and TNF-α in serum and hippocampal tissues and S100β and NSE in serum were increased in I/R and I/R-Exosome groups ( P<0.05), and no significant change was found in the parameters mentioned above in group S-Exosome ( P>0.05). The positive expression of GSDMD was significantly increased in I/R and I/R-Exosome groups ( P<0.05), no positive expression of GSDMD was found in S and S-Exosome groups ( P>0.05), and the results of immunofluorescence showed the co-localization between exosomes and microglia. Conclusions:The mechanism by which hepatic I/R induces brain injury may be related to serum exosomes-mediated microglial pyroptosis in young rats.

Effects of berberine on necroptosis of non-alcoholic fatty liver disease in mice: Relationship with AMPK/STAT6 pathway / 中华内分泌代谢杂志

Objective:To evaluate the effects of berberine on necroptosis of non-alcoholic fatty liver disease in mice and its relationship with adenosine monophosphate-activated protein kinase(AMPK)/ signal transducer and activator of transcription 6(STAT6) pathway.Methods:Twenty-five 8-week-old male C57BL/6N mice were divided into control group, steatotic liver group, berberine treatment group(200 mg·kg -1·d -1), AMPK inhibitor Compound C treatment group(0.2 mg·kg -1·d -1), and STAT6 inhibitor AS1517499 treatment group(10 mg·kg -1·d -1). After 12 weeks of intervention, the mice and liver tissue were weighed, and serum aspartate aminotransferase(AST), alanine aminotransferase(ALT), triglyceride, tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) as well as liver malondialdehyde and superoxide dismutase were measured; liver tissue HE, Masson, and oil red O staining were performed. Western blotting was used to detect the expressions of necroptosis related proteins[receptor interaction protein kinase 3(RIPK3), phosphorylated(p-) mixed lineage kinase domain-like(MLKL)], AMPK, p-AMPK, and p-STAT6. Results:Compared with control group, the steatotic liver group had higher quality of liver and liver index, and higher levels of serum AST, ALT, triglyceride, TNF-α, IL-1β, and oxidative stress( P<0.05); Liver tissue was full of cavity changes and inflammatory cell infiltration, widely distributed red lipid droplets and obvious blue fiber dyeing; The expressions of RIPK3 and p-MLKL were up-regulated ( P<0.05), but the levels of p-AMPK and p-STAT6 were relatively reduced ( P<0.05). Compared with the steatotic liver group, berberine intervention decreased liver quality and liver index, improved liver function, reduced blood lipid levels, pro-inflammatory factor expression and oxidative stress level, and significantly alleviated the degree of liver steatosis and fibrosis, the levels of RIPK3 and p-MLKL ( P<0.05), while the expressions of p-AMPK and p-STAT6 were increased significantly ( P<0.05). As compared with the berberine treatment, AMPK and STAT6 inhibitor treatment could offset the protective effect of berberine on steatotic liver, moreover, the expressions of RIPK3 and p-MLKL were increased ( P<0.05). There was no statistical difference in AMPK total protein content among the five groups ( P>0.05). Conclusion:Berberine can activate AMPK/STAT6 pathway to inhibit the necroptosis of hepatocyte, thus plays a protective role on non-alcoholic fatty liver disease in mice.

Characteristics of changes in postoperative early lung ultrasound imaging in pediatric patients undergoing living-related liver transplantation / 中华麻醉学杂志

Objective:To determine the characteristics of changes in postoperative early lung ultrasound imaging in pediatric patients undergoing living-related liver transplantation.Methods:A total of 96 pediatric patients of both sexes, aged 3-24 months, of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, with Child-Pugh grade B or C, scheduled for elective living-related liver transplantation performed with the piggy back technique, were selected.At 24 h before surgery (T 0), at 24 h after surgery (T 1) and at 72 h after surgery (T 2), the condition of lung ultrasound was recorded.Ultrasound examination was performed using Color Doppler ultrasound diagnostic instrument with a linear array probe (frequency 7-13 MHz). The lung was divided into 12 areas, and each area of both lungs was scanned successively.The lung ultrasound score in each area was recorded.The results with the highest scores were considered as the ultrasound results of this area.The general conditions of the pediatric patients were recorded and the fluid volume (input minus output) per kilogram per minute was calculated. Results:Compared with the values at T 0, the scores for lung consolidation and the pulmonary edema in lateral area were significantly increased at T 1, and the scores for lung consolidation, pulmonary edema and pleural effusion in posterior area and total score for lung consolidation, pulmonary edema and pleural effusion in each area were increased at T 1, 2 ( P<0.05). Compared with the values at T 1, the scores for lung consolidation in posterior area and total score for lung consolidation in each area were significantly decreased, and the scores for pulmonary edema in lateral and posterior areas and total score for pulmonary edema in each area were decreased at T 2 ( P<0.05), and no significant change was found in the lung ultrasound scores in anterior area at each time point ( P>0.05). Compared with the values in anterior area, the scores for lung consolidation at each time point were significantly increased, scores for pulmonary edema at T 1, 2 were increased, and score for pleural effusion at T 1 was increased in lateral area, and scores for lung consolidation, pulmonary edema and pleural effusion at each time point were increased in posterior area ( P<0.05). Compared with the values in lateral area, the scores for lung consolidation, pulmonary edema and pleural effusion at each time point were significantly increased in posterior area ( P<0.05). Conclusion:Lung consolidation, pulmonary edema and pleural effusion tend to occur in the lateral and posterior areas of the lung at 1 day after living-related liver transplantation, and the complications are severer in posterior areas.The lung consolidation in the posterior region is alleviated than that in the anterior area, and the pulmonary edema in the lateral and posterior areas is alleviated than that in the anterior area.However, all of these indexes don′t return back to their preoperative status.

Role of SIRT1/Nrf2 signaling pathway in berberine-induced reduction of renal ischemia-reperfusion injury in mice / 中华麻醉学杂志

Objective:To evaluate the role of silencing information regulator 1 (SIRT1)/nuclear factors E2-related factor2 (Nrf2) signaling pathway in berberine-induced reduction of renal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 5 groups ( n=6 each) using a random number table method: sham operation group (S group), renal I/R group (RIR group), berberine+ I/R group (B group), berberine+ I/R+ SIRT1 inhibitor EX527 group (BE group) and berberine+ I/R+ Nrf2 inhibitor ATRA group (BA group). After the right kidney was removed, the left renal artery was clamped for 45 min followed by reperfusion to establish the model of renal I/R injury.In B, BE, and BA groups, berberine 100 mg·kg -1·d -1 was given for intragastric administration at 14 days before surgery.In group BE and group BA, EX527 5 mg·kg -1·d -1 and ATRA 10 mg·kg -1·d -1 were injected intraperitoneally at 3 days before surgery, respectively.The equal volume of normal saline was given for 14 consecutive days in group S and group RIR.Blood samples were collected from orbital vein at 24 h of reperfusion for measurement of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations, for determination of the interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) contents (by enzyme-linked immunosorbent assay) and expression of SIRT1, Nrf2, apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, nucleotide-binding oligomerization domain-like receptor containing pyrin domain (NLRP3) (by Western blot) and for examination of the pathological changes of renal tubules (with a light microscope). The damage to the renal tubules was scored. Results:Compared with group S, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly increased in RIR, B, BE and BA groups, the expression of SIRT1, Nrf2, ASC, caspase-1 and NLRP3 was up-regulated in RIR, BE and BA groups, and the expression of SIRT1, Nrf2, caspase-1 and NLRP3 was up-regulated in group B ( P<0.05). Compared with group RIR, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly decreased in B, BE and BA groups, the expression of SIRT1 and Nrf2 in group B, Nrf2 and ASC in BE group and SIRT1, ASC and caspase-1 in BA group was up-regulated, and the expression of ASC, caspase-1 and NLRP3 in group B, SIRT1 and NLRP3 in BE group and Nrf2 in BA group was down-regulated ( P<0.05). Compared with group B, the serum concentrations of Cr and BUN, the contents of IL-1β and TNF-α and renal tubular injury score were significantly increased in BE and BA groups, the expression of ASC, caspase-1 and NLRP3 in BE and BA groups was up-regulated, and the expression of SIRT1 in BE and Nrf2 in BA groups was down-regulated ( P<0.05). Conclusion:SIRT1/Nrf2 signaling pathway is involved in the process of berberine-induced reduction of renal I/R, which is related to inhibiting pyroptosis in mice.

Berberine prevents steatotic liver ischemia reperfusion injury by inhibiting endoplasmic reticulum stress and autophagy / 中华器官移植杂志

Objective To explore the effect of berberine (BBR) on steatotic liver ischemia reperfusion injury and analyze the role of endoplasmic reticulum stress and autophagy .Methods Thirty-four Wistar rats were fed with a high-fat diet for 12 weeks and 2 rats were randomly selected after 8 weeks to observe pathological changes and confirm the model of steatotic liver successfully .Then before opening and closing abdominal cavity ,32 rats were divided into I/R group (normal saline was intragastrically 4 weeks before performing cold I/R treatment) ,BBR group (normal saline was replaced by BBR ,BBR was intragastrically at a dose of 300 mg·kg-1 ·d-1 weeks and others were the same as I/R group) and TG group (TG was intraperitoneally at a dose of 0 .2 mg·kg-124h pre-operation and others were the same as BBR group ) .Then the rats were sacrificed at 6h post-reperfusion .Blood samples were collected from inferior vena cava and hepatic tissues harvested .The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected ,histopathologic changes observed by Hematoxylin & Eosin (HE) staining ,oxidative stress and inflammation determined by ELISA kit and the expressions of p-PERK ,CHOP ,Bip ,LC3 ,Beclin-1 and p62 detected by Western blot .Results As compared with Sham group ,the serum levels of ALT and AST were significantly higher in I/R ,BBR and TG groups (P < 0 .05) .And hepatic histological changes were severe and oxidative stress increased in parallel with the enhancement of pro-inflammation (P< 0 .05) .In BBR group ,the level of hepatic enzymes declined ,liver injury was milder ,oxidative stress decreased and pro-inflammation was lesser compared with I/R and TG groups (P< 0 .05) .Additionally ,as compared with sham group ,the expressions of p-PERK ,CHOP ,Bip ,LC3 ,Beclin-1 and p62 were up-regulated in I/R and BBR groups (P < 0 .05) .TG group increased the levels of LC3 ,Beclin-1 and p62 (P< 0 .05) .Interestingly ,compared with I/R group ,BBR pretreatment down-regulated the expressions of p-PERK ,CHOP ,Bip ,LC3 ,Beclin-1 and p62 (P< 0 .05) .TG group had the higher expressions of LC3 ,Beclin-1 and p62 than those of BBR group (P< 0 .05) .Conclusions BBR pretreatment can protect steatotic liver ischemia reperfusion injury .And the mechanisms may be attributed to the inhibitions of endoplasmic reticulum stress and autophagy .

Effect of berberine on fatty liver ischemia-reperfusion injury in rats: the relationship with endoplasmic reticulum stress in liver tissues / 中华麻醉学杂志

Objective To evaluate the effect of berberine on fatty liver ischemia-repeffusion (I/R) injury and the relationship with endoplasmic reticulum stress in liver tissues.Methods Thirty-two male Wistar rats,aged 4 weeks,weighing 100-150 g,were divided into 4 groups (n=8 each) using a random number table method:control group (group C),fatty liver group (group FL),I/R group and berberine group (group BBR).Rats were fed a normal fat diet for 12 weeks,normal saline 3.5 ml was given intragastrically for 4 weeks starting from 9th week,and rats only underwent simple laparotomy in group C.Rats were fed a high-fat diet (45％ energy originating from fat) for 12 weeks,and the other treatments were similar to those previously described in group C.Rats were fed a high-fat diet (45％ energy originating from fat) for 12 weeks,normal saline 3.5 ml was given intragastrically for 4 weeks starting from 9th week,and then the model of liver I/R injury was established in group I/R.Rats were fed a high-fat diet (45％ energy originating from fat) for 12 weeks,berberine solution (300 mg/kg) 3.5 ml was given intragastrically for 4 weeks starting from 9th week,and then the model of liver I/R injury was established in group BBR.Hepatic ischemia was induced by clamping the portal vein,hepatic artery,right gastric vein,and supra-and inferior-hepatic vena cava to perform cold perfusion with 4 ℃ lactated Ringer's solution lasting for 30 min,followed by reperfusion.The serum triglyceride (TG) concentrations were determined after 4,8 and 12 weeks of diet.Blood samples were collected at 6 h of reperfusion for measurement of serum aspartate transminase (AST) and alanine transaminase (ALT) concentrations.Livers were removed after blood sampling at 6 h of reperfusion and liver tissues were obtained and stained with oil red O and haematoxylin and eosin for examination of pathological changes and for determination of the expression of Bip,CCAAT/enhancer-binding protein homologous protein (CHOP),protein kinase RNA-like endoplasmic reticulum kinase (PERK) and phosphorylated PERK (p-PERK) (by Western blot).p-PERK/PERK ratio was calculated.Results Compared with group C,the serum TG,ALT and AST concentrations were significantly increased,the expression of Bip and CHOP was up-regulated,p-PERK/PERK ratio was increased (P＜0.05),lipid deposition was increased,and liver steatosis was found in group FL.Compared with group FL,the serum AST and ALT concentrations were significantly increased,the expression of Bip and CHOP was up-regulated,pPERK/PERK ratio was increased (P＜0.05),and the pathological changes of liver tissues were accentuated in group I/R.Compared with group I/R,the serum TG,ALT and AST concentrations were significantly decreased,the expression of Bip and CHOP was down-regulated,p-PERK/PERK ratio was decreased (P＜ 0.05),lipid deposition was reduced,and the pathological changes of liver tissues were significantly attenuated in group BBR.Conclusion Berberine can ameliorate fatty liver I/R injury,and the mechanism may be related to inhibiting endoplasmic reticulum stress in liver tissues of rats.

Role of mammalian target of rapamycin in synaptic plasticity of entorhinal area-hippocampal formation in rats with inflammatory pain / 中华麻醉学杂志

Objective To evaluate the role of mammalian target of rapamycin (mTOR) in the synaptic plasticity of entorhinal area-hippocampal formation in rats with inflammatory pain.Methods Twenty-four healthy adult male Sprague-Dawley rats,weighing 180-240 g,were divided into 4 groups (n =6 each) by using a random number table method:control group (group C),inflammatory pain group (group IP),dimethyl sulfoxide (DMSO) group and mTOR inhibitor rapamycin group (group R).Inflammatory pain model was established by subcutaneous injection of 50 μl bee venom into the plantar surface of the left hindpaw.The equal volume of normal saline was subcutaneously injected into the plantar surface of the left hindpaw in group C.In group DMSO,2％ DMSO was administered by intragastric gavage for 3 days,1 ml per day,and the inflammatory pain model was established at 1 h after administration on 3rd day.In group R,rapamycin was administered by intragastric gavage for 3 days,1 ml per day,and the inflammatory pain model was established at 1 h after administration on 3rd day.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 h after establishing the model.The rats were sacrificed after measurement of the pain threshold,and hippocampi were removed to prepare hippocampal slices.Hippocampal CA1 region and dentate gyrus (DG region) were located with an inverted microscope.Planar microelectrode array technique was used to record the number of channels and the standardized amplitude of evoked effective field excitatory postsynaptic potentials (fEPSPs) (fEPSPs amplitude＞20％ of the baseline value) at different stimulus intensities.Results Compared with group C,MWT was significantly decreased,TWL was shortened,the number of effective fEPSP channels at different stimulus intensities was increased,and the amplitude of standardized fEPSPs in hippocampal DG and CA1 regions was increased in group IP (P＜0.05 or 0.01),and no significant change was found in the parameters mentioned above in group R (P＞0.05).Compared with group IP,MWT was significantly increased,TWL was prolonged,the number of effective fEPSP channels at different stimulus intensities was decreased,and the amplitude of standardized fEPSPs in hippocampal DG and CA1 regions was decreased in group R (P＜0.05 or 0.01),and no significant change was found in the parameters mentioned above in group DMSO (P＞0.05).Conclusion mTOR is involved in the changes in the synaptic plasticity of entorhinal areahippocampal formation in rats with inflammatory pain.

Prognosis of post-transplant diabetes mellitus in recipients with steatotic donor livers / 中华肝胆外科杂志

Objective To study the risk factors and prognosis of post-transplant diabetes mellitus (PTDM) in recipients with steatotic donor livers.Method The clinical data of 182 patients who underwent liver transplantation from donors with liver steatosis in Tianjin First Central Hospital from June 2002 to December 2010 were retrospectively analyzed.There were sixteen patients with PTDM and one hundred sixtysix without PTDM.The clinical data of these patients were compared and the risk factors were evaluated by COX regression analysis.The 1-,3-,5-year cumulative survival rates were analyzed after liver transplantation.Result The variables which included sex,pretransplant serum creatinine level,model for end-stage liver disease (MELD) score,intraoperative red blood cell transfusion,and post-transplant biliary complications were significantly different between the two groups.Multivariate Cox regression analysis showed that living-donor,pretransplant fasting blood glucose and post-transplant biliary complications could affect the survival time of patients in PTDM group.The 1-,3-and 5-year cumulative survival rates in the PTDM group were 81.3％,68.8％ and 62.5％,which were significantly lower than those in the non-PTDM group (95.2％,86.1％ and 80.7％ respectively,P ＜ 0.05).Conclusions Living-donation,pretransplant fasting blood glucose and post-transplant biliary complications had a worse prognosis in the PTDM group.A comparatively better long-term survival after liver transplantation can be achieved by reducing the risk factors and the occurrence of PTDM.

Role of autophagy in hydrogen-induced inhibition of apoptosis in hippocampal neurons in a rat model of orthotopic liver transplantation / 中华麻醉学杂志

Objective To evaluate the role of autophagy in hydrogen-induced inhibition of apoptosis in hippocampal neurons in a rat model of orthotopic liver transplantation (OLT).Methods Fifty-six pathogen-free healthy adult male Sprague-Dawley rats,aged 8-10 weeks,weighing 220-250 g,were used in the study.Thirty-two rats were selected and assigned into 4 groups (n =8 each) using a random number table:sham operation group (group S),OLT group,hydrogen-rich saline group (group HS) and chloroquine group (group CQ).The other 24 rats severed as the donors.In group S,laparotomy was performed,and the related blood vessels were isolated.The model of OLT was established in OLT,HS and CQ groups.In group OLT,normal saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group HS,hydrogen-rich saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group CQ,autophagy inhibitor chloroquine 60 mg/kg was injected intraperitoneally at 1 h before establishment of the model,and the other treatments were similar to those previously described in group HS.At 6 h of reperfusion,the rats were sacrificed and hippocampi were isolated for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity,for pathological examination (with light microscope),and for detection of cell apoptosis (by TUNEL staining) and expression of autophagy-and apoptosis-related proteins caspase-3,cytochrome c (Cyt c),microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),Beclin-1 and p53 in hippocampal tissues (by Western blot analysis).Apoptosis index (AI) was calculated.Results Compared with group S,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3,Cyt c,LC3 Ⅱ,Beclin-1 and p53 was up-regulated in OLT,HS and CQ groups (P＜0.05).Compared with group OLT,the MDA content and AI were significantly decreased,the SOD activity was increased,the expression of caspase-3 and Cyt c was down-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was up-regulated in group HS (P＜0.05).Compared with group HS,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3 and Cyt c was up-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was down-regulated in group CQ (P＜0.05).Conclusion The mechanism by which hydrogen inhibits apoptosis in hippocampal neurons is related to promotion of autophagy in a rat model of OLT.

Comparison of myocardial injury in pediatric patients undergoing living-donor liver transplantation performed under propofol-versus sevoflurane-based anesthesia / 中华麻醉学杂志

Objective To compare the severity of myocardial injury in pediatric patients undergoing living-donor liver transplantation (LDLT) performed under propofol-versus sevoflurane-based anesthesia.Methods Forty American Society of Anesthesiologists physical status Ⅲ or Ⅳ pediatric patients of both sexes,aged 5-13 months,weighing 5-12 kg,scheduled for elective LDLT under general anesthesia,were divided into 2 groups (n =20 each) using a random number table:propofol-based anesthesia group (group P) and sevoflurane-based anesthesia group (group S).Anesthesia was induced with intravenously injected midazolam 0.15 mg/kg,fentanyl 2-5 μg/kg and cisatracurium 0.15 mg/kg.Anesthesia was maintained by Ⅳ infusion of propofol 9-15 mg · kg-1 · h-1 (in group P),continuous inhalation of 2.6％-4.0％ sevoflurane (in group S),intermittent Ⅳ boluses of fentanyl 1-3 μg/kg and Ⅳ infusion of cisatracurium 1-2 μg · kg-1 · min-1.At 5 min of anesthesia induction,30 min of anhepatic phase,3 h of neohepatic stage and the end of surgery,blood samples were taken from the central vein for determination of concentrations of cardiac troponin Ⅰ and creatine kinase-MB in serum and concentrations of tumor necrosis factor-alpha,interleukin-6 and high-mobility group box 1 protein in serum (by enzyme-linked immunosorbent assay).The occurrence of adverse cardiovascular events (hypertension or hypotention,myocardial ischemia and ventricular premature beat) was recorded during surgery.Results There was no significant difference in concentrations of tumor necrosis factor-alpha,interleukin-6,high-mobility group box 1 protein,cardiac troponin Ⅰ and creatine kinase-MB in serum at each time point or incidence of adverse cardiovascular events between the two groups (P＞0.05).Conclusion The severity of myocardial injury is comparable in pediatric patients undergoing LDLT performed under propofol-versus sevoflurane-based anesthesia.

Effect of dexmedetomidine on myocardial injury in pediatric patients undergoing living-related liver transplantation / 中华麻醉学杂志

Objective To evaluate the effect of dexmedetomidine on myocardial injury in pediatric patients undergoing living-related liver transplantation. Methods Fifty-eight pediatric patients of both sexes,aged 5-20 months,weighing 4.5-15.0kg,of American Society of Anesthesiologists physical status Ⅲ or Ⅳ,scheduled for elective living-related liver transplantation,were divided into 2 groups(n=29 each)using a random number table:control group(group C)and dexmedetomidine group(group D).In group D,dexmedetomidine was intravenously infused at a dose of 0.5 μg/kg over 10min starting from the time point immediately before skin incision,followed by an infusion of 0.8 μg·kg-1·h-1 until the end of surgery. The equal volume of normal saline was given instead in group C. Immediately before skin incision(baseline,T0),at 10min of anhepatic phase(T1),at 30min of neohepatic phase(T2)and at the end of surgery(T3),blood samples were obtained from the central vein for determination of serum cardiac troponin I(cTnI),lactate dehydrogenase(LDH),alpha-hydroxybutyrate dehydrogenase(α-HBDH),interleukin-6(IL-6)and IL-10 concentrations. The changing rate of serum cTnI concentrations were calculated at T2. The occurrence of myocardial ischemia and ventricular premature beat and requirement for dopamine were recorded during surgery. Results Compared with the baseline at T0,the serum concentrations of cTnI,LDH and α-HBDH were significantly increased at T2,3,and the serum concentrations of IL-6 and IL-10 were increased at T1-3 in both groups(P0.05).Conclusion Dexmedetomidine can attenuate the myocardial injury to some extent in pediatric patients undergoing living-related liver transplantation.

Role of SIRT1/FoxO3α signaling pathway in berberine pretreatment-induced reduction of hypoxia/ reoxygenation-caused injury to hepatic parenchymal cells / 中华麻醉学杂志

Objective To evaluate the role of silent information regulator fac tor 2-related enzyme 1 (SIRT1)/Forkhead Box O3 (FoxO3a) signaling pathway in berberine pretreatment-induced reduction of hypoxia/reoxygenation (H/R)-caused injury to hepatic parenchymnal cells.Methods Hepatic parenchymal cells obtained from AML12 mice were cultured and seeded in 6-well plates (2 ml/well) and in 96-well plates (200 μl/well) at the density of l×l06 cells/ml.The cells were divided into 4 groups (n=36 each)using a randomn number table:control group (group C),group H/R,berberine pretreatment group (group BP) and SIRT1-siRNA group (group SS).The cells were cultured in normal culture atmosphere (5％ CO2-21％ O2-74％ N2) in group C.In H/R,BP and SS groups,the cells were exposed to hypoxic air (5％ CO2-1％ O2-94％ N2) for 12 h,followed by 6 h reoxygenation in normal culture atmosphere (5％ CO2-21％ O2-74％ N2).In group SS,small interference RNA targeting SIRT1 (SIRT1-siRNA) was added to the culture medium at 24 h prior to hypoxia.Berberine (final concentration 5 μmol/L) was added at 2 h prior to hypoxia in BP and SS groups.At the end of reoxygenation,the cell viability was measured by methyl thiazolyl tetrazolium assay,the malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined using enzyme-linked immunosorbent assay,cell apoptosis was detected by flow cytometry,the expression of SIRT1 and FoxO3α was detected by Western blot,and the acetylation of FoxO3α was measnred by using immunoprecipitation.Apoptotic rate was calculated.Results Compared with group C,the cell viability was significantly decreased,the MDA content was increased,the SOD activity was decreased,apoptotic rate was increased,the expression of SIRT1 and ratio of FoxO3α expression in nucleus/in cytoplasma were increased,and the acetylation of FoxO3α in the nucleus was increased in H/ R,BP and SS groups (P＜ 0.05).Compared with group H/R,the cell viability was significantly increased,the MDA content was decreased,the SOD activity was increased,apoptotic rate was decreased,the expression of SIRT1 and ratio of FoxO3α expression in nucleus/in cytoplasma were increased,and the acetylation of FoxO3α in the nucleus was increased in group BP (P＜0.05).Compared with group BP,the cell viability was significantly decreased,the MDA content was increased,the SOD activity was decreased,apoptotic rate was increased,the expression of SIRT1 and ratio of FoxO3α expression in nucleus/in cytoplasma were decreased,and the acetylation of FoxO3α in the nucleus was decreased in group SS (P＜O.05).Conclusion The mechanism by which berberine pretreatment attenuates H/R-caused injury to hepatic parenchymal cells is related to promotion of SIRT1 expression in cells and inhibition of FoxO3α acetylation in the nucleus.

Role of Nrf2 signaling pathway in attenuation of ischemia injury by hydrogen-rich University of Wisconsin solution during cold storage of rat donor kidneys / 中华麻醉学杂志

Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway in attenuation of ischemia injury by hydrogen-rich University of Wisconsin (HRUW) solution during cold storage of rat donor kidneys.Methods Forty healthy adult male Wistar rats,weighing 200-250 g,were divided into 4 groups (n =10 each) using a random number table:control group (group C),University of Wisconsin (UW) solution group (group UW),HRUW solution group (group HRUW) and Nrf2 inhibitor all-trans retinoic acid (ATRA) group (group ATRA).ATRA 7 mg/kg was intraperitioneally injected once a day for 2 consecutive days,kidneys were isolated and underwent cold storage at 8 h after the last administration,and kidneys were stored in HRUW solution for 48 h at 4 ℃C in group ATRA.In UW and HRUW groups,the equal volume of normal saline was intraperitioneally injected instead,and isolated kidneys were stored in UW solution and HRUW solution for 48 h at 4 ℃C,respectively.Kidney specimens were obtained for microscopic examination and for determination of tumor necrosis factoralpha (TNF-α),interleukin-lbeta (IL-1β3),high-mobility group box 1 protein (HMGB1),IL-10 and 8-iso-prostaglandin F2α (8-iso-PGF2α) contents (by enzyme-linked immunosorlbent assay),superoxide dismutase (SOD) and catalase (CAT) activities (using spectrophotometry),and expression of Nrf2,heme oxygenase-1 (HO-1),Bcl-2,Bax and caspase-3 in renal tissues (by using Western blot).The damage to the renal tubules was scored.Results Compared with group C,renal tubular damage scores were signifieantly increased,TNF-α,IL-1β,HMGB1 and 8-iso-PGF2α contents were increased,IL-10 contents were decreased,the expression of Nrf2 and HO-1 was up-regulated,SOD and CAT activities were decreased,the expression of Bcl-2 was down-regulated,and the expression of Bax and caspase-3 was upregulated in group UW (P＜0.05 or 0.01).Compared w,ith group UW,renal tubular damage scores were significantly decreased,TNF-α,IL-1β,HMGB1 and 8-iso-PGF2α contents were decreased,IL-10 contents were increased,the expression of Nrf2 and HO-1 was up-regulated,SOD and CAT activities were increased,the expression of Bcl-2 was up-regulated,and the expression of Bax and caspase-3 was down-regulated in group HRUW,and the expression of Nrf2 and Bcl-2 was up-regulated (P＜0.05),and no significant change was found in the other parameters in group ATRA (P＞0.05).Compared witb group HRUW,renal tubular damage seores were significantly increased,TNF-α,IL-1β,HMGB1 and 8-iso-PGF2α contents were increased,IL-10 contents were decreased,the expression of HO-1 and Bcl-2 was down-regulated,SOD and CAT activities were decreased,and the expression of Bax and caspase-3 was up-regulated in group ATRA.Conclusion HRUW solution reduces inflammatory responses,oxidative damage and cell apoptosis during cold storage of rat donor kidneys,and the mechanism by which HRUW solution attenuates ischemia injury is related to activation of Nrf2 signaling pathway.

Role of Nrf2/HO-1 signaling pathway in reduction of acute kidney injury following orthotopic liver transplantation by hydrogen-rich saline in rats / 中华麻醉学杂志

Objective To investigate the role of nuclear factor erythroid 2-related factor 2 (Nrf2) / heme oxygenase-1 (HO-1) signaling pathway in reduction of acute kidney injury following orthotopic liver transplantation (OLT) by hydrogen-rich saline in rats.Methods Thirty-two healthy adult male SpragueDawley rats,weighing 220-250 g,were randomly assigned into 4 groups (n =8 each) using a random number table:sham operation group (S group),OLT group,hydrogen-rich saline group (HS group),and all-trans retinoic acid (ATRA) group.Laparotomy was performed,and the related blood vessels were isolated in group S.The model of orthotopic autologous liver transplantation was established in OLT,HS and ATRA groups.Normal saline and hydrogen-rich saline 6 ml/kg were injected through the inferior vena cava at 5 min before the portal vein was clamped in OLT and HS groups,respectively.In group ATRA,Nrf2 inhibitor ATRA 7 mg/kg was injected intraperitoneally once a day for 2 consecutive days,the model of orthotopic autologous liver transplantation was established at 16 h after the last injection of ATRA,and the other treatments were similar to those previously described in group HS.At 6 h of reperfusion,blood samples were collected for determination of serum blood urea nitrogen (BUN),creatinine (Cr),interleukin10 (1L-10) and tumor necrosis factor-alpha (TNF-α) concentrations.After blood sampling,the lungs were removed for determination of malondialdehyde (MDA) content,superoxide dismutase (SOD) activity,expression of HO-1,Bcl-2 and Bax mRNA (by using real-time reverse transcriptase polymerase chain reaction),and HO-1 protein expression in lung tissues (by Western blot) and for microscopic examination.The damage to the renal tubules was scored.Results Compared with group S,the serum BUN,Cr and TNF-α concentrations were significantly increased,the serum IL-10 concentrations were decreased,the MDA content and renal tubular damage score were increased,the SOD activity was decreased,and the expression of HO-1 protein and mRNA,and Bcl-2 and Bax mRNA was up-regulated in group OLT (P＜ 0.05).Compared with group OLT,the serum BUN,Cr and TNF-α concentrations were significantly decreased,the serum IL-10 concentrations were increased,the MDA content and renal tubular damage score were decreased,the SOD activity was increased,the expression of HO-1 protein and mRNA and Bcl-2 mR-NA was up-regulated,and the expression of Bax mRNA was down-regulated in group HS (P＜0.05).Compared with group HS,the serum BUN,Cr and TNF-α concentrations were significantly increased,the serum IL-10 concentrations were decreased,the MDA content and renal tubular damage score were increased,the SOD activity was decreased,the expression of HO-1 protein and mRNA and Bcl-2 mRNA was down-regulated,and the expression of Bax mRNA was up-regulated in group ATRA (P＜0.05).Conclusion The mechanism by which hydrogen-rich saline reduces acute kidney injury following OLT is probably associated with activation of Nrf2/HO-1 signaling pathway in rats.

Prognostic value of serum cardiac troponin Ⅰ in acute lung injury after living-donor liver transplantation for children with biliary atresia / 中华器官移植杂志

Objective To evaluate the impact of cardiac troponin Ⅰ (cTnI) on acute lung injury in pediatric living donor liver transplant children with biliary atresia.Methods The clinical data of 112 pediatric living donor liver transplant recipients with biliary atresia in Tianjin First Central Hospital from February 2011 to September 2015 were retrospectively reviewed.Fifty-five recipients with cTnI ≥0.07 μg/L served as high-cTnI group and 57 recipients with cTnI group ＜0.07μg/L as normalcTnI group.The clinical data between two groups were compared and the association between serum cTnI level and acute lung injury after living-donor liver transplantation was evaluated by logistic regression analysis.Results The percentage of acute lung injury after pediatric living donor liver transplantation in high-cTnI group and normal-cTnI group was 31.6％ and 9.1％,respectively.Intratransplant cTnI ≥0.07μg/L (OR =4.489,confidence interval 1.170-17.226) was the risk factor for acute lung injury after transplantation.The value of cTnI showed the positive correlation with preoperative PELD scores (OR =4.489,confidence interval 1.170-17.226).Conclusions Intratransplant cTnI level was the significant prognostic risk factor in acute lung injury after pediatric living-donor liver transplantation for children with biliary atresia.The cTnI level was associated with preoperative PELD scores.

Effect of dexmedetomidine on kidney injury induced by liver ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the effect of dexmedetomidine on kidney injury induced by liver ischemia-reperfusion (I/R) in rats.Methods Twenty-four healthy male Sprague-Dawley rats,weighing 220-250 g,aged 8-10 weeks,were randomly divided into 3 groups (n=8 each) using a random number table:sham operation group (group S);liver I/R group (group I/R);dexmedetomidine group (group D).In group I/R,liver I/R model was established by clamping the portal vein,hepatic artery,supra-and infra-hepatic vena cava for 40 min,followed by 6 h of reperfusion in anesthetized rats.In group D,dexmedetomidine 100 μg/kg was injected intraperitoneally at 30 min before skin incision.The equal volume of normal saline was given instead of dexmedetomidine in S and I/R groups.At 6 h of reperfusion,blood samples were collected from the infra-hepatic vena cava for determination of blood urea nitrogen (BUN) and creatinine (Cr) concentrations (by automatic biochemical analyzer) and tumor necrosis factor-alpha (TNFα) and interleukin-10 (IL-10) concentrations in serum (by enzyme-linked immunosorbent assay).After blood sampling,the rats were sacrificed,and kidneys were harvested for examination of histopathological changes (with light microscope) and for determination of malondialdehyde (MDA) content (using thiobarbituric acid method) and superoxide dismutase (SOD) activity (by xanthine oxidase method),expression of activated caspase-3 (by immuno-histochemistry),and apoptotic cells (using TUNEL).Apoptotic rate was calculated.Results Compared with group S,the serum BUN,Cr and TNF-α concentrations were significantly increased,the concentration of serum IL-10 was decreased,the MDA content and apoptotic rate were increased,the SOD activity was decreased,and the expression of activated caspase-3 was up-regulated in I/R and D groups (P＜0.05).Compared with group I/R,the serum BUN,Cr and TNF-α concentrations were significantly decreased,the concentration of serum IL-10 was increased,MDA content and apoptotic rate were increased,the SOD activity was decreased,the expression of activated caspase-3 was down-regulated (P＜0.05),and the histopathological changes of renal tissues were attenuated in group D.Conclusion Dexmedetomidine can reduce kidney injury induced by liver I/R in rats,and the mechanism is probably related to inhibition of inflammatory responses,lipid peroxidation and cell apoptosis.

Berberine preconditioning protects against hepatic cold ischemia reperfusion injury through the modulation of PI3K/Akt/mTOR signaling pathway / 中华器官移植杂志

Objective To confirm the protective effect of berberine (BBR) on cold ischemia reperfusion (I/R)-induced liver injury and to show whether the hepatic protection conferred by BBR involves the activation of phosphatidylinositol 3 kinase (PI3K) / protein kinase B (Akt)/mammalian target of rapamycin(mTOR) signal pathway.Method Adult male Sprague-Dawley rats were assigned randomly to four groups:BBR group (BBR was intragastrically administered at a dose of 100 mg·kg-1 · d-1 2 weeks before hepatic cold I/R treatment),dimethyl sulfoxide (DMSO) group (BBR was replaced by DMSO,and others were the same as BBR group),I/R group (BBR was replaced by normal saline,and others were the same as BBR group) and sham group (normal saline was administered 2 weeks before opening and closing abdomen treatment).Then the rats were sacrificed at 3,6,and 24 h after reperfusion.The liver function,oxidative stress level,apoptosis rate,and the expression of PI3K/Akt/mTOR related pathway proteins were assayed.Result As compared with sham group,the I/R-induced liver tissue displayed severe lobular distortion with widespread necrosis,high level of oxidative stress and apoptosis rate.As compared with I/R group,BBR dramatically attenuated the histopathologic damage,restored the liver function and decreased the oxidative stress level.Simultaneously,BBR significantly ameliorated the apoptosis by decreasing the apoptosis rate,increasing the Bcl-2/Bax ratio and inhibiting caspase-3 activity in rats subjected to hepatic I/R.The expression of p-Akt was effectively upregulated with the inhibited expression of p-mTOR.Conclusion Our result provides robust in vivo evidence that BBR can prevent I/R-induced oxidative stress and apoptosis.The mechanisms involved can be attributed to the activation of P]3K/Akt/mTOR signal pathway.

Effect of berberine pretreatment on hypoxia/reoxygenation-induced apoptosis in human renal tubular epithelial cells / 中华麻醉学杂志

Objective To investigate the effect of berberine pretreatment on hypoxia/reoxygenation (H/R)-induced apoptosis in human renal tubular epithelial cells.Methods The human renal tubular epithelial cells were cultured and seeded in culture dishes (2 ml/dish) or 96-well plates (200 μl/well) with the density of 1 ×106/ml.The cells were then randomly divided into 4 groups (n =30 each):normal control group (group C),berberine group (group B),H/R group and H/R + berberine group (H/R + B group).In groups B and H/R + B,berberine 10 μmol/L was added to the culture medium and the cells were incubated for 2 h.Groups H/R and H/R + B were then exposed to 94％ N2-5％ CO2-1％ O2 for 24 h followed by 3 h reoxygenation.The cell viability,apoptotic rate and level of malondialdehyde (MDA) and superoxide dismutase (SOD) were detected.The expression of caspase-3,activated caspase-3,cytochrome c,glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) was determined.The ratio of Bax/Bcl-2 was calculated.Results Compared with group C,the cell viability,SOD activity and caspase-3 expression were significantly decreased,the apoptotic rate,Bax/Bcl-2 ratio and concentration of MDA were increased,and the expression of activated caspase-3,cytochrome c,GRP78 and CHOP was up-regulated in groups H/R and H/R + B (P ＜ 0.05).Compared with group H/R,the cell viability,SOD activity and caspase-3 expression were significantly increased,the apoptotic rate,Bax/Bcl-2 ratio and concentration of MDA were decreased,and the expression of activated caspase-3,cytochrome c,GRP78 and CHOP was down-regulated in group H/R + B (P ＜ 0.05).Conclusion Berberine pretreatment can inhibit H/R-induced apoptosis in human renal tubular epithelial cells,and inhibition of mitochondrial stress pathway and endoplasmic reticulum stress pathway is involved in the mechanism.

In vitro study on Coxsackie virus B3 infection on islet cells / 中华微生物学和免疫学杂志

Objective To study the effect of Coxsackie virus B3 (CVB3) infection on islet cells in vitro, and to explore the mechanism of islet cells caused by CVB3. Methods Bone marrow mesenchymal stem cells( BMSCs) were separated from the bone marrow and cultured. Then they were induced to differentiate into islet-like cells using nicotinamide and mercaptoethanol. Differentiated cells were detected by morphology , special staining and RT-PCR. Observe CVB3 infection on islet cells under inverse microscope and detect the specific gene fragment by RT-PCR. Results BMSCs showed half suspended shape and gathered to form a cluster after induction. Cells became red brown by dithizone specific staining. RT-PCR also proved the existence of mRNA expressing insulin. Infected islet cells appeared typical pathological changes like shrinks, refraction decreases. RT-PCR detected the desired specific gene fragment of 299 bp in infected islet cells. Conclusion CVB3 can directly injury islet cells, and damage the function of islet cells of secreting insulin.