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The review summarizes the risk factors, diagnostic criteria and perioperative control strategies of acute kidney injury in pediatric liver transplantation, aiming to provide rationales for proper managements.
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The report described one case of vascular paralysis syndrome during kidney transplantation to provide references for clinical practice.After intraoperative opening of kidney artery and vein, the recipient developed vascular paralysis syndrome.However, the efficacy is not obvious after dosing of norepinephrine.After an intravenous infusion of methylene blue, the recipient has a successful removal of tracheal intubation and recovered well.
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Objective:To evaluate the effect of stroke volume variation(SVV) goal-directed fluid therapy on postoperative pulmonary complications(PPCs) after pediatric living donor liver transplantation.Methods:One hundred and twenty pediatric patients undergoing pediatric living-donor liver transplantation(all diagnosed with congenital biliary atresia) were divided into 2 groups( n=60 each) using the random number table method: control group and SVV group. Intraoperative fluid management was guided by central venous pressure and mean arterial pressure in control group, while by SVV combined with cardiac output in SVV group. Intraoperative circulation, fluid intake and usage of vasoactive drug were recorded. Central venous blood samples were collected to determine the concentrations of serum Clara cell 16 kDa protein, interleukin-6, and tumor necrosis factor-alpha before anesthesia(T 0), at the end of anhepatic phase(T 1), at 3 h of neohepatic phase(T 2), at the end of surgery(T 3) and at 24 h after operation(T 4). Pulmonary ultrasonography was performed before surgery, at the end of surgery and at 1, 3 and 7 days after surgery. The pediatric patients were followed up for 1 week after surgery to record the PPCs, including acute lung injury, pulmonary infection, pulmonary atelectasis, pleural effusion and acute respiratory distress syndrome. Results:Compared with control group, the incidence of PPCs, acute lung injury and pulmonary infection was significantly decreased, the pulmonary ultrasound score was decreased at the end of surgery and at 1, 3 and 7 days after surgery, the usage of intraoperative dobutamine was increased, the duration of postreperfusion syndrome was shortened, the fluid intake and epinephrine usage were reduced, and the serum Clara cell 16 kDa protein, tumor necrosis factor-alpha and interleukin-6 concentrations were decreased at T 1-T 4 in SVV group( P<0.05). Conclusions:SVV goal-directed fluid management can reduce the development of PPCs in pediatric living donor liver transplantation.
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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.
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Objective:To compare the myocardial protection in pediatric patients undergoing living-donor liver transplantation (LDLT) performed under propofol- versus desflurane-based anesthesia. Methods:Sixty American Society of Anesthesiologists Physical Status classification Ⅲ or Ⅳ pediatric patients of both sexes, aged 5-24 months, weighing 5-15 kg, scheduled for elective LDLT under general anesthesia, were divided into 2 groups ( n=30 each) using a random number table method: propofol group (group P) and desflurane anesthesia group (group D). During anesthesia maintenance, propofol 5-10 mg·kg -1·min -1 was intravenously infused in group P, desflurane 0.65 MAC-1.30 MAC was inhaled in group D. At 5 min after induction of anesthesia, at 1 h of reperfusion, at the end of surgery, at 1, 2, 3, 7 and 14 days after surgery, and on the day of discharge, the concentrations of serum high-sensitivity cardiac troponin I, creatine kinase isoenzyme, N-terminal pro-B-type natriuretic peptide were determined by enzyme-linked immunosorbent assay, the occurrence of nausea and vomiting, agitation, and shivering, postoperative tracheal extubation time, intensive care unit stay time, and postoperative length of hospital stay were recorded within 24 h after surgery. Results:Compared with group P, the concentrations of serum high-sensitivity cardiac troponin I and creatine kinase isoenzyme were significantly decreased after surgery, the extubation time and intensive care unit stay time were shortened ( P<0.05), and no significant change was found in serum N-terminal pro-B-type natriuretic peptide concentrations, postoperative length of hospital stay and incidence of postoperative adverse effects at each time point in group D ( P>0.05). Conclusions:Desflurane has better myocardial protection than propofol in pediatric patients undergoing LDLT, which is helpful for early prognosis.
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Objective:To evaluate the effects of inhalation of high-concentration hydrogen on acute kidney injury (AKI) and mitochondrial dynamics in septic mice.Methods:One hundred and twenty-eight male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=32 each) using a random number table method: sham operation group (group Sham), sham operation + hydrogen group (group Sham+ H), sepsis AKI group, and sepsis AKI+ hydrogen group (group S-AKI+ H). A mouse model of sepsis-induced AKI was developed by cecal ligation and puncture in anesthetized animals. In Sham+ H and S-AKI+ H groups, 67% H 2+ 33% O 2 was inhaled for 1 h starting from 1 and 6 h after sham operation or developing the model, respectively. Twenty mice were selected to observe the survival at 7 days after developing the model. At 24 h after developing the model, blood samples were collected for determination of serum BUN and Cr concentrations (by colorimetric analysis), and renal tissues were obtained for determination of the contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and high mobility group protein B1 (HMGB1) (by enzyme-linked immunosorbent assay), activities of superoxide dismutase (SOD) and catalase (CAT) (by spectrophotometry) and expression of dynamin-related protein 1 (Drp1) and mitofusin 2 (Mfn2) (by Western blot). The damage to the renal tubules was scored after HE staining. Results:Compared with Sham group, the survival rate was significantly decreased, the serum BUN and Cr concentrations, renal tubular damage score and contents of TNF-α, IL-1β and HMGB1 were increased, the activities of SOD and CAT were decreased, the expression of Drp1 was up-regulated, and the expression of Mfn2 was down-regulated in S-AKI group ( P<0.05), and no significant change was found in the parameters mentioned above in Sham+ H group ( P>0.05). Compared with S-AKI group, the survival rate was significantly increased, the serum BUN and Cr concentrations, renal tubular injury score and contents of TNF-α, IL-1β and HMGB1 were decreased, the activities of SOD and CAT were increased, the expression of Drp1 was down-regulated, and the expression of Mfn2 was up-regulated in S-AKI+ H group ( P<0.05). Conclusions:Inhalation of high-concentration hydrogen can alleviate AKI in septic mice, and the mechanism may be related to inhibition of renal mitochondrial fission and promotion of mitochondrial fusion.
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Objective:To evaluate the effects of inhaling high concentration hydrogen on myocardial injury and mitochondrial biogenesis in septic mice.Methods:One hundred and twenty-eight clean-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=32 each) using a random number table method: sham operation group (group Sham), sham operation + hydrogen group (group Sham+ H), sepsis group (group Sep), and sepsis+ hydrogen group (group Sep+ H). The sepsis model was developed by cecal ligation and puncture in anesthetized animals. In Sham+ H and Sep+ H groups, 67% H 2 was inhaled for 1 h starting from 1 and 6 h after operation, respectively. Twenty mice in each group were randomly selected to observe the survival conditions at 7 days after operation. Blood samples were taken from the remaining mice at 24 h after operation for determination of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), cardiac troponin I (cTnI) and creatine kinase isoenzyme (CK-MB) (by enzyme-linked immunosorbent assay), for examination of the pathological changes of myocardial tissues (by HE staining), and for determination of the mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry), ATP content (by luciferase assay), and expression of myocardial peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), nuclear respiratory factor 2 (NRF2) and mitochondrial transcription factor A (TFAM) (by Western blot). Results:Compared with Sham group, the survival rate was significantly decreased, the serum concentrations of TNF-α, IL-1β, cTnI and CK-MB and pathological score were increased, the MMP and content of ATP in myocardial mitochondria were decreased, and the expression of PGC-1α, NRF2 and TFAM in myocardial tissues was down-regulated in Sep group ( P<0.05), and no significant change was found in the parameters mentioned above in Sham+ H group ( P>0.05). Compared with group Sep, the survival rate was significantly increased, the serum concentrations of TNF-α, IL-1β, cTnI and CK-MB and pathological score were decreased, the MMP and content of ATP in myocardial mitochondria were increased, and the expression of PGC-1α, NRF2 and TFAM in myocardial tissues was up-regulated in group Sep+ H ( P<0.05). Conclusions:Inhaling high concentration hydrogen can attenuate sepsis-induced myocardial injury in mice, and the mechanism may be related to promotion of mitochondrial biosynthesis and improvement in mitochondrial function.
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Objective:To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on postoperative acute lung injury (ALI) in the pediatric patients undergoing living-related liver transplantation.Methods:Sixty pediatric patients of either sex, aged 4-24 months, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, with New York Heart Association (NYHA) class Ⅰ or Ⅱ, with Child-Pugh B or C, scheduled to undergo elective left external lobe piggyback living-related liver transplantation, were divided into 2 groups ( n=30 each) using a computer-generated table of random numbers: control group (group C) and TEAS group (group T). In group T, bilateral Zusanli (ST36), Neiguan (PC6), and Feishu (BL13) acupoints were stimulated with disperse-dense waves at the initial intensity of 0.5 mA and frequency of 2/15 Hz, the current intensity was gradually increased until local slight muscle shaking appeared, and continuous stimulation lasted for 30 min at a 30-min interval (a cycle) until the end of operation. TEAS was performed for 30 min at the same time every day up to 1 week after surgery. Stimulus locations in group C were selected at 0.5 cm lateral to the acupoints, and the electrodes with inert medium were attached to the location, with no effective current output from acupuncture treatment instrument. The peak inspiratory pressure, plateau pressure, and pulmonary compliance were recorded before skin incision (T 0), at 30 min after portal vein occlusion (T 1), at 1 h after portal vein opening (T 2), at the end of operation (T 3), and the difference between peak inspiratory pressure and plateau pressure was calculated. Blood samples from the jugular vein were collected at T 0-3 to determine the levels of plasma club cell protein 16 (CC16), surfactant protein D (SP-D), soluble receptor for advanced glycation end products (sRAGE), tumor necrosis factor-alpha (TNF-α), and interleukin-10 (IL-10) by enzyme-linked immunosorbent assay. Blood samples from the radial artery were collected at T 0-3 for blood gas analysis, PaO 2 and A-aDO 2 were recorded, and oxygenation index (OI) and respiratory index (RI) were calculated. The indwelling time of postoperative tracheal tube and length of ICU stay were also recorded. The lung injury was assessed and scored using ultrasound at 48 h after surgery. The occurrence of ALI within 1 week after operation was also recorded. Results:Compared with baseline at T 0, OI was significantly decreased, RI was increased, and plasma IL-10 concentrations were increased at T 2, 3, and the plasma concentrations of TNF-α, CC16, sRAGE and SP-D were increased at T 1-3 in both groups ( P<0.05). Compared with group C, OI was significantly increased, RI was decreased, the plasma concentrations of sRAGE were decreased, and the plasma concentrations of IL-10 were increased at T 2, 3, and the concentrations of plasma TNF-α, CC16 and SP-D were decreased at T 1-3, the indwelling time of postoperative tracheal tube and length of ICU stay were shortened, the ultrasound score of lung injury was decreased ( P<0.05), and no significant change was found in the incidence of ALI in group T ( P>0.05). Conclusions:TEAS can alleviate ALI in the pediatric patients after living-related liver transplantation.
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Objective:To identify the risk factors for acute lung injury (ALI) after pediatric living donor liver transplantation (LDLT) and evaluate the predictive value.Methods:The pediatric patients (all diagnosed with congenital biliary atresia) who underwent parental liver transplantation in our center from January to December 2021 were selected. Perioperative data were obtained through the electronic medical record system, and the pediatric patients were divided into non-ALI group and ALI group according to whether ALI occurred or not at 1 week after surgery. The factors of which P values were less than 0.05 between groups would enter the multivariate logistic regression analysis to stratify the risk factors for ALI after pediatric LDLT, and the value of the risk factors in predicting intraoperative ALI was evaluated using the receiver operating characteristic curve. Results:A total of 140 pediatric patients were enrolled in the analysis, and the incidence of ALI was 30.7%. The results of the multivariate logistic regression analysis showed that preoperative pediatric end-stage liver disease score, preoperative serum NT-pro-BNP concentrations, intraoperative volume of fluid transfused, and duration of postreperfusion syndrome were independent risk factors for ALI after LDLT in pediatric patients ( P<0.05). The area under the receiver operating characteristic curve of the preoperative N-terminal pro-brain natriuretic peptide(NT-pro-BNP) concentration in predicting postoperative ALI was 0.737 ( P<0.001), with a cutoff value of 222.1 ng/L, sensitivity of 0.628, and specificity of 0.732. Conclusions:Preoperative pediatric end-stage liver disease score, serum NT-pro-BNP concentrations, intraoperative volume of fluid transfused, and duration of postreperfusion syndrome are independent risk factors for ALI after LDLT in pediatric patients; preoperative serum NT-pro-BNP concentrations can effectively predict the development of ALI after pediatric LDLT surgery.
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BACKGROUND@#Red-cell transfusion is critical for surgery during the peri-operative period; however, the transfusion threshold remains controversial mainly owing to the diversity among patients. The patient's medical status should be evaluated before making a transfusion decision. Herein, we developed an individualized transfusion strategy using the West-China-Liu's Score based on the physiology of oxygen delivery/consumption balance and designed an open-label, multicenter, randomized clinical trial to verify whether it reduced red cell requirement as compared with that associated with restrictive and liberal strategies safely and effectively, providing valid evidence for peri-operative transfusion.@*METHODS@#Patients aged >14 years undergoing elective non-cardiac surgery with estimated blood loss > 1000 mL or 20% blood volume and hemoglobin concentration <10 g/dL were randomly assigned to an individualized strategy, a restrictive strategy following China's guideline or a liberal strategy with a transfusion threshold of hemoglobin concentration <9.5 g/dL. We evaluated two primary outcomes: the proportion of patients who received red blood cells (superiority test) and a composite of in-hospital complications and all-cause mortality by day 30 (non-inferiority test).@*RESULTS@#We enrolled 1182 patients: 379, 419, and 384 received individualized, restrictive, and liberal strategies, respectively. Approximately 30.6% (116/379) of patients in the individualized strategy received a red-cell transfusion, less than 62.5% (262/419) in the restrictive strategy (absolute risk difference, 31.92%; 97.5% confidence interval [CI]: 24.42-39.42%; odds ratio, 3.78%; 97.5% CI: 2.70-5.30%; P <0.001), and 89.8% (345/384) in the liberal strategy (absolute risk difference, 59.24%; 97.5% CI: 52.91-65.57%; odds ratio, 20.06; 97.5% CI: 12.74-31.57; P <0.001). No statistically significant differences were found in the composite of in-hospital complications and mortality by day 30 among the three strategies.@*CONCLUSION@#The individualized red-cell transfusion strategy using the West-China-Liu's Score reduced red-cell transfusion without increasing in-hospital complications and mortality by day 30 when compared with restrictive and liberal strategies in elective non-cardiac surgeries.@*TRIAL REGISTRATION@#ClinicalTrials.gov, NCT01597232.
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Humans , Adult , Postoperative Complications , Erythrocyte Transfusion/adverse effects , Blood Transfusion , Hospitals , Hemoglobins/analysisABSTRACT
Many neutralizing antibodies (nAbs) elicited to ancestral SARS-CoV-2 through natural infection and vaccination generally have reduced effectiveness to SARS-CoV-2 variants. Here we show therapeutic antibody ADG20 is able to neutralize all SARS-CoV-2 variants of concern (VOCs) including Omicron (B.1.1.529) as well as other SARS-related coronaviruses. We delineate the structural basis of this relatively escape-resistant epitope that extends from one end of the receptor binding site (RBS) into the highly conserved CR3022 site. ADG20 can then benefit from high potency through direct competition with ACE2 in the more variable RBS and interaction with the more highly conserved CR3022 site. Importantly, antibodies that are able to target this site generally neutralize all VOCs, albeit with reduced potency against Omicron. Thus, this highly conserved and vulnerable site can be exploited for design of universal vaccines and therapeutic antibodies.
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Objective:To evaluate the role of nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy in intestinal ischemia-reperfusion (I/R) injury in mice and the relationship with ferroptosis.Methods:Thirty SPF-grade healthy male C57BL/6 mice, aged 8-10 weeks, weighing 21-25 g, were divided into 5 groups ( n=6 each) using a random number table method: sham operation group (Sham group), intestinal I/R group (I/R group), intestinal I/R + NCOA4 silencing group (I/R+ NCOA4 shRNA group), intestinal I/R + autophagy inhibitor 3-methyladenine (3-MA) group (I/R+ 3-MA group), and intestinal I/R + NCOA4 silencing + autophagy activator rapamycin (RAPA) group (I/R+ NCOA4 shRNA + RAPA group). The intestinal I/R injury model was developed by clamping the superior mesenteric artery for 45 min followed by reperfusion in anesthetized animals.At 2 weeks before developing the model, AAV-NCOA4-shRNA 1×10 11 vp was injected via the tail vein in group I/R+ NCOA4 shRNA and group I/R+ NCOA4 shRNA+ RAPA, and AAV shCtrl (adenovirus control) 1 x10 11 vp was injected in Sham, I/R and I/R+ 3-MA groups.Rapamycin 4 mg/kg was intraperitoneally injected once a day starting from 7 days before developing the model in group I/R+ NCOA4 shRNA+ RAPA.In group I/R+ 3-MA, 3-MA 10 mg/kg was intraperitoneally injected at 1 h before developing the model.The animals were sacrificed at 2 h of reperfusion, and intestinal tissues were obtained for determination of contents of malondialdehyde (MDA), glutathione (GSH) and Fe 2+ (by colorimetry), contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) (by enzyme-linked immunosorbent assay), and expression of NCOA4, microtubule-associated protein 1 light chain 3 (LC3), long-chain fatty acyl-CoA synthase 4 (ACSL4), ferritin heavy chain 1 (FTH1) and glutathione peroxidase 4 (GPX4) (by Western blot). The LC3Ⅱ/LC3Ⅰ ratio was calculated.Intestinal damage was assessed and scored according to Chiu. Results:Compared with group Sham, the Chiu′s score and contents of MDA, Fe 2+ , TNF-α and IL-1β were significantly increased, GSH content was decreased, the expression of NCOA4 and ACSL4 was up-regulated, LC3Ⅱ/LC3Ⅰ ratio was increased, and the expression of GPX4 and FTH1 was down-regulated in group I/R ( P<0.05). Compared with group I/R, the Chiu′s score and contents of MDA, Fe 2+ , TNF-α and IL-1β were significantly decreased, and GSH content was increased in I/R+ NCOA4 shRNA and I/R+ 3-MA groups, the expression of NCOA4 and ACSL4 was significantly down-regulated, and the expression of GPX4 and FTH1 was up-regulated in group I/R+ NCOA4 shRNA ( P<0.05), and ACSL4 expression was significantly down-regulated, LC3Ⅱ/LC3Ⅰ ratio was decreased, and the expression of GPX4 and FTH1 was up-regulated in group I/R+ 3-MA ( P<0.05). Conclusions:NCOA4-mediated ferritinophagy can promote ferroptosis, which is involved in the pathophysiological mechanism of intestinal I/R injury in mice.
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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.
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Objective:To evaluate the role of microRNA-10a (miRNA-10a) in renal ischemia-reperfusion (I/R) injury in mice and the relationship with transforming growth factor beta1 (TGF-β 1)/Smad2 signaling pathway. Methods:Twenty-four SPF healthy male adult C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups ( n=6 each) by the random number table method: sham operation group (S group), renal I/R group (IR group), renal I/R plus miRNA-10a antagonist group (I group), and renal I/R+ miRNA-10a agonist group (M group). The mouse model of renal I/R was developed by clamping the left renal pedicle for 45 min followed by reperfusion in anesthetized animals.miRNA-10a antagonist and agonist 20 nmol were injected via the tail vein once every 24 h for 3 consecutive days starting from 72 h before surgery in group M and group I, respectively, while the equal volume of normal saline was given instead in S and IR groups.Blood samples were collected from the orbital vein at 24 h of reperfusion to determine the concentrations of the serum creatinine (Scr) and blood urea nitrogen (BUN). Then the mice were sacrificed, and the kidney tissues were taken for determination of the malondialdehyde (MDA) content and superoxide dismutase (SOD) activity, contents of interleukin-1 beta (IL-β) and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay), and expression of TGF-β 1 and phosphorylated Smad2 (p-Smad2) (by Western blot) and for microscopic examination of the pathological changes.The damage to the renal tubules was scored. Results:Compared with group S, the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly increased in IR, I and M groups, and the activity of SOD was significantly decreased, and the expression of TGF-β 1 and p-Smad2 was up-regulated in IR and M groups ( P<0.05). Compared with group IR, the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly decreased, the activity of SOD was increased, and the expression of TGF-β 1 and p-Smad2 was down-regulated in group I, and the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly increased, the activity of SOD was decreased, and the expression of TGF-β 1 and p-Smad2 was up-regulated in group M ( P<0.05). Compared with group I, the concentrations of Scr and BUN, contents of MDA, IL-1β and TNF-α in renal tissues and renal tubular damage score were significantly increased, the activity of SOD was decreased, and the expression of TGF-β 1 and p-Smad2 was up-regulated in group M ( P<0.05). Conclusions:miRNA-10a is involved in the process of renal I/R injury and is related to activation of TGF-β 1/Smad2 signaling pathway in mice.
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Objective:To evaluate the role of nicotinamide adenine dinucleotide-dependent deacetylase Sirtuin3 (Sirt3) expression in peritoneal macrophages in dexmedetomidine-induced inhibition of inflammatory responses in septic mice.Methods:Sixty-four healthy male C57BL/6 mice, aged 7 weeks, weighing about 20 g, were divided into 4 groups ( n=16 each) using a random number table method: sham operation group (S group), sepsis group (Sep group), dexmedetomidine group (DEX group), and dexmedetomidine plus Sirt3 inhibitor 3-TYP group (TYP group). Sepsis was induced by cecal ligation and puncture.In group DEX, normal saline 0.25 ml was intraperitoneally injected at 1 h before development of the model, and 30 min later dexmedetomidine 50 μg/kg (in 0.25 ml of normal saline) was intraperitoneally injected.In group S and group Sep, the equal volume of normal saline was intraperitoneally injected at 1 h and 30 min before development of the model, respectively.In group S, laparotomy and cecal extraction were performed without ligation and perforation.Peritoneal lavage was obtained at 24 h after operation, and peritoneal macrophages were cultured for 1 h. The expression of Sirt3, interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) mRNA in peritoneal macrophages was detected using quantitative real-time polymerase chain reaction.Western blot was used to detect the expression of Sirt3 in peritoneal macrophages.The concentrations of IL-1β and TNF-α in peritoneal fluid were measured by enzyme-linked immunosorbent assay.The 10-day survival of mice in each group was observed, and the survival rates were calculated. Results:Compared with group S, the expression of Sirt 3 protein and mRNA in peritoneal macrophages was significantly down-regulated, the expression of IL-1β and TNF-α mRNA was up-regulated, the concentrations of IL-1β and TNF-α in peritoneal lavage were increased, and the 10-day survival rates were decreased in Sep, DEX and TYP groups ( P<0.05). Compared with group Sep, the expression of Sirt3 protein and mRNA in peritoneal macrophages was significantly up-regulated, the expression of IL-1β and TNF-α mRNA was down-regulated, the concentrations of IL-1β and TNF-α in the peritoneal lavage were decreased, and the 10-day survival rate was increased in group DEX ( P<0.05). Compared with group DEX, the expression of Sirt3 protein and mRNA and IL-1β and TNF-α mRNA in peritoneal macrophages was significantly up-regulated, the concentrations of IL-1β and TNF-α in peritoneal lavage were increased, and the 10-day survival rate was decreased in group TYP ( P<0.05). Conclusions:Up-regulation of Sirt3 expression in peritoneal macrophages is involved in the process of dexmedetomidine-induced inhibition of inflammatory responses in septic mice.
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Objective:To evaluate the role of ecto-5′-nucleotidase (CD73) in endogenous protective mechanism of hepatic ischemia-reperfusion (I/R) injury in mice and the relationship with transforming growth factor beta1 (TGF-β 1)/Smad3 signaling pathway. Methods:Twenty-four SPF healthy male C57BL/6 mice, aged 8-10 weeks, weighing 20-23 g, were divided into 3 groups ( n=8 each) by the random number table method: sham operation group (S group), hepatic I/R group (IR group) and hepatic I/R plus CD73 specific inhibitor group (APCP group). The hepatic hilum was only exposed but not occluded in group S. The hepatic portal was occluded for 30 min followed by reperfusion to develop the model of hepatic I/R in anesthetized animals in group IR.CD73-specific inhibitor APCP 40 mg/kg was intraperitoneally injected, and 10 min later hepatic I/R was performed.Orbital venous blood samples were collected at 6 h of reperfusion for determination of serum alanine transaminase (ALT) and aspartate transaminase (AST) concentrations.Then the mice were sacrificed, and liver tissues were obtained for determination of the expression of CD73, TGF-β 1 and phosphorylated Smad3 (p-Smad3) (by Western blot), contents of interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay), and content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) (with a visible spectrophotometer) and for microscopic examination of the pathological changes of liver tissues (with a light microscope). Results:Compared with group S, the concentrations of AST and ALT in serum and contents of IL-1β, TNF-α and MDA in liver tissues were significantly increased, the activity of SOD was decreased, and the expression of CD73, TGF-β 1 and p-Smad3 was up-regulated in IR and APCP groups ( P<0.05). Compared with group IR, the concentrations of AST and ALT in serum and contents of IL-1β, TNF-α and MDA in liver tissues were significantly increased, the activity of SOD was decreased, and the expression of CD73, TGF-β 1 and p-Smad3 in liver tissues was down-regulated in group APCP ( P<0.05). The pathological changes of liver tissues were accentuated in group APCP as compared with group IR. Conclusions:CD73 is involved in the process of endogenous protective mechanism of hepatic I/R injury in mice, which may be related to the regulation of TGF-β 1/Smad3 signaling pathway.
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Objective:To explore the effect of intensive insulin therapy on hemodynamics and cardiac function in organ donors.Methods:A total of 60 organ donors were randomly divided into two groups of intensive insulin therapy(IIT)and control(30cases each group). Blood glucose was adjusted at 6.2~10.0 mmol/L in control group and 4.4~6.1 mmol/L in IIT group.Blood glucose and insulin dosage during maintenance were recorded.Cardiac function values as well as serum inflammatory factor concentrations at admission and during donation were compared between two groups.Results:During maintenance, blood glucose was significantly lower in IIT group than that in control group [(5.1±0.6)vs(8.2±1.5)mmol/L, P<0.05] and insulin dosage was higher than that in control group [(9.5±3.2)vs(5.8±1.5)U/h, P<0.05]. As compared with control group, cardiac cycle efficiency(CCE), maximal rate of elevated pressure(DP/DT max)and left ventricular ejection fraction(LVEF)in were significantly higher in IIT group than those of control group.And serum cardiac troponin I(cTnI), N-terminal B-type natriuretic peptide(NT-Pro-BNP), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and high mobility group box-1 protein(HMGB1)as well as vasoactive-inotropic score(VIS)were significantly lower than those in control group( P<0.05). As compared with control group, cardiac donation rate of IIT group was significantly higher(30% vs 16.7%, P<0.05). Conclusions:Intensive insulin therapy and blood glucose control may blunt inflammatory response in organ donors, lessen myocardial injury and myocardial depression, stabilize hemodynamics and boost the rate of cardiac donation.
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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.
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Objective:To evaluate the effect of hepatic ischemia-reperfusion (I/R) rats-derived exosomes on microglial pyroptosis.Methods:Twenty clean-grade healthy male Sprague-Dawley rats, aged 2-3 weeks, weighing 20-50 g, were divided into 2 groups ( n=10 each) using a random number table method: sham operation group (group S) and hepatic I/R group (group I/R). The serum of rats in S group and I/R group was collected, and exosomes were isolated from the sera using differential centrifugations.Microglial cells were co-cultured with PKH26-labeled exosomes for 6 h. The intake of exosomes in microglial cells was determined using immunofluorescence staining.Primary microglial cells were seeded onto 6-well culture plates at a density of 5×10 5 cells/ml and were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), 10 7 cells/ml I/R-exosomes treated group (group 10 7), 10 8 cells/ml I/R-exosomes treated group (group 10 8), and 10 9 cells/ml I/R-exosomes treated group (group 10 9). Microglia in each group were co-cultured with the corresponding concentration of I/R-exosomes for 6 h. The expression of NOD-like receptor family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein (ASC), cleaved-caspase-1 and gasdermin-D (GSDMD) was detected using Western blot.Primary microglial cells were divided into 3 groups ( n=24 each) by a random number table method: control group (group C), sham operation-exosomes treated group (group S-exosome) and I/R-exosomes treated group (group I/R-exosome). In S-exosome group and I/R-exosome group, exosomes 10 8 cells/ml in S group and I/R group were given, respectively, to incubate cells for 6 h. The expression of NLRP3, ASC and GSDMD mRNA was determined by real-time polymerase chain reaction, and the levels of interleukin-18 (IL-18), IL-1β and tumor necrosis factor-alpha (TNF-α) in the cell culture supernatant were measured by enzyme-linked immunosorbent assay. Results:The results from immunofluorescence staining showed that I/R-exosomes colocalized with microglia.The 10 8 cells/ml I/R-exosomes and 10 9 cells/ml I/R-exosomes up-regulated the expression of NLRP3, ASC, GSDMD and cleaved-caspase-1 in microglial cells ( P<0.01). Compared with group C and group S-exosome, the expression of NLRP3, ASC and GSDMD mRNA in microglial cells was up-regulated, and the levels of IL-18, IL-1β and TNF-α in the supernatant were elevated in group I/R-exosome ( P<0.01). Conclusions:Hepatic I/R rats-derived exosomes can promote microglial pyroptosis.
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SARS-CoV-2 Beta variant of concern (VOC) resists neutralization by major classes of antibodies from non-VOC COVID-19 patients and vaccinated individuals. Here, serum of Beta variant infected patients revealed reduced cross-neutralization of non-VOC virus. From these patients, we isolated Beta-specific and cross-reactive receptor-binding domain (RBD) antibodies. The Beta-specificity results from recruitment of novel VOC-specific clonotypes and accommodation of VOC-defining amino acids into a major non-VOC antibody class that is normally sensitive to these mutations. The Beta-elicited cross-reactive antibodies share genetic and structural features with non-VOC-elicited antibodies, including a public VH1-58 clonotype targeting the RBD ridge independent of VOC mutations. These findings advance our understanding of the antibody response to SARS-CoV-2 shaped by antigenic drift with implications for design of next-generation vaccines and therapeutics. One sentence summarySARS-CoV-2 Beta variant elicits lineage-specific antibodies and antibodies with neutralizing breadth against wild-type virus and VOCs.
