The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic aci...The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid(5-ALA).Furthermore,this study aimed to explore the underlying mechanism of the protective effects of 5-ALA.First,we collected and stored mouse testicular fragments in different media,including Hank’s balanced salt solution(HBSS;n=5),Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F12;n=5),and alpha-minimum essential medium(αMEM;n=5).Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group(P<0.05)and theαMEM group(P<0.01).Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA(0[control],1 mmol l−1,2 mmol l−1,and 5 mmol l−1)to determine the most effective concentration of 5-ALA.The 2 mmol l−15-ALA group(n=3)presented the highest positive rate of spermatogonial stem cells compared with those in the control,1 mmol l−1,and 5 mmol l−15-ALA groups.Finally,the tissue fragments were preserved in HBSS with control(n=3)and 2 mmol l−15-ALA(n=3)under low-temperature conditions.A comparative analysis was performed against fresh testes(n=3)to elucidate the underlying mechanism of 5-ALA.Gene set enrichment analysis(GSEA)for WikiPathways revealed that the p38 mitogen-activated protein kinase(MAPK)signaling pathway was downregulated in the 2 mmol l−15-ALA group compared with that in the control group(normalized enrichment score[NES]=−1.57,false discovery rate[FDR]=0.229,and P=0.019).In conclusion,these data suggest that using 2 mmol l−15-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.展开更多
Background:The use of grafts from donation after circulatory death(DCD)overcomes the inadequate donor organ supply.Our team developed a transportable dual hypothermic oxygenated machine perfusion(DHOPE)device,which in...Background:The use of grafts from donation after circulatory death(DCD)overcomes the inadequate donor organ supply.Our team developed a transportable dual hypothermic oxygenated machine perfusion(DHOPE)device,which initiates DHOPE at a recipient center to reduce static cold storage(SCS)time and the risk of graft failure in DCD liver transplantation.Methods:Six porcine livers per group with 30 min of warm ischemia exposure were preserved via SCS or DHOPE for 6 h and then reperfused for 12 h with whole blood to mimic transplantation.Hepatocellular and biliary function and injury were assessed in perfusate and bile samples.Molecular biomarkers and histology were compared between groups.Results:Reperfusion portal vein pressure,in a flow-constant manner,and alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP)and gamma-glutamyltransferase(γ-GGT)release were significantly lower in the DHOPE group than in the SCS group at all time points.Higher bile production paralleled the lower levels of ALP andγ-GGT in the DHOPE group.The DHOPE group secreted more total bilirubin(TBIL)in bile,resulting in decreased TBIL in the perfusate,and livers preserved with DHOPE exhibited better cholangiocellular function.Furthermore,improvements in hypoxia,the inflammatory response,cell-free microRNAs and energy metabolism were observed in the DHOPE group.There were fewer apoptotic cells and TGF-β1-positive cells in the liver parenchyma and extrahepatic bile duct in the DHOPE group than in the SCS group.Conclusions:This study demonstrates the efficacy of local 4℃DHOPE to protect porcine liver grafts from30-min warm ischemia damage.展开更多
AIM: To optimize the perfusates used for hypothermicmachine perfusion(HMP).METHODS: Sprague-Dawley rats were assigned randomly to three groups(n = 12 per group) that received either saline, University of Wisconsin col...AIM: To optimize the perfusates used for hypothermicmachine perfusion(HMP).METHODS: Sprague-Dawley rats were assigned randomly to three groups(n = 12 per group) that received either saline, University of Wisconsin coldstorage solution(UW) or histidine-tryptophan-ketoglutarate solution(HTK) as the perfusate. Each group was divided into two subgroups: static cold storage(SCS) and HMP(n = 6 per subgroup). The liver graft was retrieved according to the method described by Kamada. For the SCS group, the graft was directly placed into cold perfusate(0-4?℃) for 6 h after liver isolation while the portal vein of the graft was connected to the perfusion machine for the HMP group. Then the perfusates were collected at different time points for analysis of aspartate aminotransferase(AST), alanine transaminase(ALT) and lactate dehydrogenase(LDH) levels. Liver tissues were obtained for evaluation of histology, dry/wet weight(D/W) ratio, and malondialdehyde(MDA) and adenosine-triphosphate(ATP) levels. The portal vein pressure and velocity were monitored in real time in all HMP subgroups.RESULTS: Comparison of HMP and SCS: Regardless of the perfusate, HMP improved the architecture of donor graft in reducing the congestion around sinusoids and central vein and maintaining sinusoid lining in morphology; HMP improved liver function in terms of ALT, AST and LDH, especially during the 3-6 h period(SCS vs HMP using saline: ALT3, 225.00 ± 105.62 vs 49.50 ± 18.50, P = 0.047; LDH3, 1362.17 ± 563.30 vs 325.75 ± 147.43, P = 0.041; UW: LDH6, 2880.14 ± 948.46 vs 2135.00 ± 174.27, P = 0.049; HTK, AST6, 307.50 ± 52.95 vs 185.20 ± 20.46, P = 0.041); HMP decreased MDA level(saline, 2.79 ± 0.30 vs 1.09 ± 0.09, P = 0.008; UW, 3.01 ± 0.77 vs 1.23 ± 0.68, P = 0.005; HTK, 3.30 ± 0.52 vs 1.56 ± 0.22, P = 0.006). Comparison among HMP subgroups: HTK showed less portal vein resistance than UW and saline(vs saline, 3.41 ± 0.49 vs 5.00 ± 0.38, P < 0.001; vs UW, 3.41 ± 0.49 vs 4.52 ± 0.63, P = 0.007); UW reduced edema most efficiently(vs saline, 0.68 ± 0.02 vs 0.79 ± 0.05, P = 0.013), while HTK maintained ATP levels best(vs saline, 622.60 ± 29.11 vs 327.43 ± 44.66, P < 0.001; vs UW, 622.60 ± 29.11 vs 301.80 ± 37.68, P < 0.001).CONCLUSION: HMP is superior to SCS in maintaining both architecture and function of liver grafts. Further, HTK was found to be the optimal perfusate for HMP.展开更多
Oxidative stress and apoptosis are the key factors that limit the hypothermic preservation time of donor hearts to within 4–6 h.The aim of this study was to investigate whether the histone deacetylase 3(HDAC3)inhibit...Oxidative stress and apoptosis are the key factors that limit the hypothermic preservation time of donor hearts to within 4–6 h.The aim of this study was to investigate whether the histone deacetylase 3(HDAC3)inhibitor RmGodFyPn9 a6 m6 i cc opualrda mpreotteercst daugrianing srt ecpaerrdfiuasci oinn juwreyr ei nedvuacleuda tebyd.pTrholeo nexgperde shsyipoont haenrdm pich opsrpehsoerryvlaattiioonn.leRvaet lsh eoaf rtms awmemrea lihayn-??STE20-like kinase-1(Mst1)and Yes-associated protein(YAP)were determined by western blotting.Cell apoptosis was measured by the terminal deoxynucleotidyl-transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)method.Addition of RGFP966 in Celsior solution significantly inhibited cardiac dysfunction induced by hypothermic preservation.RGFP966 inhibited the hypothermic preservation-induced increase of the phosphorylated(p)-Mst1/Mst1 and p-YAP/YAP ratios,prevented a reduction in total YAP protein expression,and increased the nuclear YAP protein level.Verteporfin(VP),a small molecular inhibitor of YAP–transcriptional enhanced associate domain(TEAD)interaction,partially abolished the protective effect of RGFP966 on cardiac function,and reduced lactate dehydrogenase activity and malondialdehyde content.RGFP966 increased superoxide dismutase,catalase,and glutathione peroxidase gene and protein expression,which was abolished by VP.RGFP966 inhibited hypothermic preservation-induced overexpression of B-cell lymphoma protein 2(Bcl-2)-associated X(Bax)and cleaved caspase-3,increased Bcl-2 mRNA and protein expression,and reduced cardiomyocyte apoptosis.The antioxidant and anti-apoptotic effects of RGFP966 were cancelled by VP.The results suggest that supplementation of Celsior solution with RGFP966 attenuated prolonged hypothermic preservation-induced cardiac dysfunction.The mechanism may involve inhibition of oxidative stress and apoptosis via inactivation of the YAP pathway.展开更多
Using deep hypothermic circulatory arrest, thoracic aorta diseases and complex heart diseases can be subjected to corrective procedures. However, mechanisms underlying brain protection during deep hypothermic circulat...Using deep hypothermic circulatory arrest, thoracic aorta diseases and complex heart diseases can be subjected to corrective procedures. However, mechanisms underlying brain protection during deep hypothermic circulatory arrest are unclear. After piglet models underwent 60 minutes of deep hypothermic circulatory arrest at 14°C, expression of microRNAs(miRNAs) was analyzed in the hippocampus by microarray. Subsequently, TargetScan 6.2, RNA22 v2.0, miRWalk 2.0, and miRanda were used to predict potential targets, and gene ontology enrichment analysis was carried out to identify functional pathways involved. Quantitative reverse transcription-polymerase chain reaction was conducted to verify miRNA changes. Deep hypothermic circulatory arrest altered the expression of 35 miRNAs. Twenty-two miRNAs were significantly downregulated and thirteen miRNAs were significantly upregulated in the hippocampus after deep hypothermic circulatory arrest. Six out of eight targets among the differentially expressed miRNAs were enriched for neuronal projection(cyclin dependent kinase, CDK16 and SLC1 A2), central nervous system development(FOXO3, TYRO3, and SLC1 A2), ion transmembrane transporter activity(ATP2 B2 and SLC1 A2), and interleukin-6 receptor binding(IL6 R)– these are the key functional pathways involved in cerebral protection during deep hypothermic circulatory arrest. Quantitative reverse transcription-polymerase chain reaction confirmed the results of microarray analysis. Our experimental results illustrate a new role for transcriptional regulation in deep hypothermic circulatory arrest, and provide significant insight for the development of miRNAs to treat brain injuries. All procedures were approved by the Animal Care Committee of Xuanwu Hospital, Capital Medical University, China on March 1, 2017(approval No. XW-INI-AD2017-0112).展开更多
AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex...AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex vivo by hypothermic machine perfusion(HMP).METHODS Eighteen young(4 mo old) and 18 aged(17 mo old)healthy male SD rats were selected and randomly divided into three groups: control group, UW solution perfusion group(UWP), and UW solution with metformin perfusion group(MUWP). Aspartate aminotransferase(AST), alanine aminotransferase(ALT), lactate dehydrogenase(LDH), interleukin-18(IL-18), and tumor necrosis factor-alpha(TNF-α) in the perfused liquid were tested. The expression levels of AMPK and endothelial nitric oxide synthase(e NOS) in liver sinusoidal endothelial cells were also examined.Additionally, microscopic evaluation of the harvested perfused liver tissue samples was done. RESULTS AST, ALT, LDH, IL-18 and TNF-α levels in the young and aged liver-perfused liquid were, respectively,significantly lower in the MUWP group than in the UWP group(P < 0.05), but no significant differences were found between the young and aged MUWP groups.Metformin increased the expression of AMPK and e NOS protein levels, and promoted the extracellular release of nitric oxide through activation of the AMPK-e NOS mediated pathway. Histological examination revealed that in the MUWP group, the extent of liver cells and tissue damage was significantly reduced compared with the UWP group.CONCLUSION The addition of metformin to the UW preservative solution for ex vivo HMP can reduce rat liver injury during cold ischemia, with significant protective effects on livers, especially of aged rats.展开更多
The technique for bloodless hepatic resection using the total hepatic vascular isolation under the normothermic or hypothermic perfusion was reported to deal with the large liver tumor involving in the liver hilum,the...The technique for bloodless hepatic resection using the total hepatic vascular isolation under the normothermic or hypothermic perfusion was reported to deal with the large liver tumor involving in the liver hilum,the main hepatic veins or the retrohepatic vena cava.The original Heaney's and Fortner's methods were modified so that the technique could be simpler and more practicable to perform otherwise hazardous liver resection.During the past 4 year,major hepatic resection with the normothermic or hypothermic total vascular exclusion technique was successfully performed on 19 patients with liver tumors in our department.Among the 19 cases,16 underwent hepatic resection with the normothermic selective total vascular exclusion(extended right lobectomy in 5 cases,extended left lobectomy in 3 cases;right lobectomy in 5 cases;central segmentectomy in 3 cases)and 3 with the total vascular isolation and in situ cold perfusion(extended left lobectomy in 2 case,extended right lobectong in 1case).We believe that the technique of normothermic vascular exclusion may be indicated to deal with the lesion close to the hepatic veins and the retrohepatic vena cava.However,for more complicated hepatic resection,the hypothermic perfusion technique should be considered to prolong the safety of ischemic tune of the liver.The preliminary experience in the clinical application using the above technique is reported.展开更多
BACKGROUND Persistent left superior vena cava(PLSVC),a relatively rare thoracic vascular malformation,can inconvenience perfusionists and operators when encountered during deep hypothermic circulatory arrest(DHCA).CAS...BACKGROUND Persistent left superior vena cava(PLSVC),a relatively rare thoracic vascular malformation,can inconvenience perfusionists and operators when encountered during deep hypothermic circulatory arrest(DHCA).CASE SUMMARY Herein,we describe the case of a patient with concurrent giant aortic arch aneurysm,aortic stenosis,and PLSVC.To treat these conditions,we performed right hemiarch and aortic valve replacements under DHCA.Notably,we applied“bilateral superior vena cava retrograde cerebral perfusion(RCP)”for cerebral protection,which significantly optimized the surgical procedure and reduced the risk of postoperative complications.The patient was discharged 14 d after surgery with no complications.CONCLUSION Surgical intervention for PLSVC under DHCA can be performed using the bilateral superior vena cava RCP approach.展开更多
BACKGROUND Hypothermic machine perfusion(HMP)has demonstrated benefits in terms of early kidney transplant function compared to static cold storage.While longer preservation times have shown detrimental effects,a prev...BACKGROUND Hypothermic machine perfusion(HMP)has demonstrated benefits in terms of early kidney transplant function compared to static cold storage.While longer preservation times have shown detrimental effects,a previous paired study indicated that longer pump times(the second kidney in a pair)might lead to improved outcomes.AIM To revisit the prior paired study's somewhat unexpected results by reviewing our program's experience.METHODS A total of 61 pairs of transplant recipients who received kidneys from the same donor(2012-2021)were analyzed.Patients were divided into two groups depending on whether they were transplanted first(K1)or second(K2).Therefore,the patients in each pair had identical donor characteristics,except for time on the pump.Statistical analyses included Kaplan-Meyer analysis and paired tests,including McNemar's test,student's paired t-test,or Wilcoxon's test,as appropriate.RESULTS The two groups of recipients had similar demographics(age,body mass index,diabetes,time on dialysis,sensit-ization and retransplants).Cold ischemic times for K1 and K2 were 8.9(95%CI:7.9,9.8)and 14.7 hours(13.7,15.8)(P<0.0001),respectively.Overall,K2 had a higher rate of freedom from biopsy-proven acute rejection at 1 year(P=0.015).Delayed graft function was less common in K2,12/61(20%)than in K1,20/61(33%)(P=0.046).Finally,K2 showed a higher graft survival than K1(P=0.023).CONCLUSION Our results agree with a previous study that suggested possible advantages to longer pump times.Both studies should encourage further research into HMP's potential anti-inflammatory effect.展开更多
Background:Liver transplantation(LT)is a crucial treatment for end-stage liver disease,but the limited organ supply has led to the use of extended criteria donors(ECD).The implementation of dynamic preservation techni...Background:Liver transplantation(LT)is a crucial treatment for end-stage liver disease,but the limited organ supply has led to the use of extended criteria donors(ECD).The implementation of dynamic preservation techniques like hypothermic oxygenated machine perfusion(HOPE)is crucial in improving outcomes for ECD grafts.However,graft contamination and infection are a concern.This study aimed to evaluate the risk of infections within 10 days from LT between HOPE and static cold storage(SCS)groups and postoperative complications.Methods:A retrospective cohort study was conducted,including LT recipients transplanted at a single-center from March 2016 to June 2023.Patients were divided into HOPE and SCS groups,and propensity score matching was used to select comparable cohorts.Data on patient and donor characteristics were analyzed.Results:After propensity score matching,a cohort of 370(HOPE,n=185;SCS,n=185)patients was selected for analysis.The study found no significant differences in the rate of clinically relevant infections,microbiological positive samples,or donor-derived infections within 10 days between HOPE and SCS groups.Postoperative outcomes,as well as patient and graft survival,were also similar between the two groups.The study showed that HOPE is a feasible and safe approach,with a comparable risk of infection.Conclusions:The study results indicate that HOPE use in LT does not increase the risk of infection and is associated with similar patient and graft survival outcomes compared to SCS.These findings confirm the safety and efficacy of HOPE in LT and its potential to expand the donor pool without compromising recipient outcomes.展开更多
Cell sheet(CS)-based approaches hold significant potential for tissue regeneration,relying on the extracellular matrix(ECM)for success.Like in native tissues,the ECM provides structural and biochemical support for cel...Cell sheet(CS)-based approaches hold significant potential for tissue regeneration,relying on the extracellular matrix(ECM)for success.Like in native tissues,the ECM provides structural and biochemical support for cellular homeostasis and function.Effective preservation strategies that maintain ECM integrity are critical to enhance the therapeutic potential of CS-based approaches.While cryogenic and hypothermic preservation methods offer potential solutions,their impact on CS ECM structure is not fully understood.Therefore,a comprehensive analysis of the ECM of hASCs CS following cryogenic and hypothermic preservation for 3 and 7 days,was conducted.Although proteomic analysis indicated that cryopreservation had no significant effect on the overall composition of the ECM,it induced significant ECM structural alterations,particularly disrupting collagen organization,which was not observed following hypothermic preservation.These structural changes were accompanied by alterations in mechanical properties,including a reduction in elastic modulus.In contrast,hypothermic preservation maintained ECM integrity and mechanical properties similar to the control.The notable ECM structural changes following cryogenic preservation can potentially impact cellular behavior,including adhesion,proliferation,and differentiation,thereby affecting the efficacy of CS therapies in vivo.This suggests that hypothermia may offer a promising alternative to cryopreservation for preserving CS integrity and functionality.展开更多
Background:Ischemia reperfusion injury(IRI)is a major contributing factor to organ damage in liver transplantation(LT)impacting donor organ quality and patient survival.IRI-inflicted graft injury can be reduced by usi...Background:Ischemia reperfusion injury(IRI)is a major contributing factor to organ damage in liver transplantation(LT)impacting donor organ quality and patient survival.IRI-inflicted graft injury can be reduced by using hypothermic oxygenated machine perfusion(HOPE)as a preservation strategy instead of static cold storage(SCS).The endothelial glycocalyx is highly sensitive to IRI and its degradation during graft preservation and reperfusion was previously associated with inferior postoperative outcome after LT.Here,we aimed to measure glycocalyx degradation during and after HOPE in order to evaluate its potential for viability-assessment during machine perfusion and outcome prediction in patients undergoing LT.Methods:Glycocalyx degradation was quantified via enzyme-linked immunoassay(ELISA)for its main component syndecan-1(Sdc-1)in serum of 40 patients undergoing LT after HOPE.In addition,Sdc-1 was evaluated at multiple time points during HOPE.Patients were followed up for 3.5 years to assess postoperative complications including morbidity,the development of early allograft dysfunction(EAD)and graft survival.Results:Liver grafts which later developed EAD showed significantly higher Sdc-1 concentrations after 60 min of HOPE compared to grafts exhibiting normal postoperative function(P=0.02).Receiver operating characteristic analysis revealed a strong predictive potential with an area under the curve of 0.73.A cut-off at 808 ng/mL Sdc-1 at 60 min of HOPE allowed identification of a high-risk group with an incidence of EAD of 66.7%.Sdc-1 concentrations increased during all types of HOPE but were significantly higher in HOPE versus dual HOPE(D-HOPE)after 120 min of perfusion(P=0.02).Conclusions:Sdc-1 evaluated at 60 min during HOPE allows prediction of EAD after LT.Accordingly,Sdc-1 should be considered a potential additional biomarker for viability assessment during HOPE.展开更多
Background:Vascular resistance and flow rate during hypotherrnic machine perfusion (HMP)of kidneys is correlated with graft function. We aimed to determine the effects of increasing HMP pressure versus maintaining the...Background:Vascular resistance and flow rate during hypotherrnic machine perfusion (HMP)of kidneys is correlated with graft function. We aimed to determine the effects of increasing HMP pressure versus maintaining the initial pressure on kidney transplantation outcomes. Methods:We retrospectively reviewed the data of 76 primary transplantation patients who received HMP-preserved kidneys from 48 donors after cardiac death between September 1,2013,and August 31,2015.HMP pressure was increased from 30 to 40mmHg (1mmHg =0.133kPa)in kidneys with poor flow and/or vascular resistance (increased pressure [IP]group;36 patients);otherwise,the initial pressure was maintained (constant pressure group;40 patients).Finally,the clinical characteristics and transplantation outcomes in both groups were assessed. Results:Delayed graft function (DGF)incidence,1-year allograft,patient survival,kidney function recovery time,and serum creatinine level on day 30 were similar in both groups,with improved flow and resistance in the IP group.Among patients with DGF,kidney function recovery time and DGF duration were ameliorated in the IP group.Multivariate logistic regression analysis revealed that donor hypertension (odds ratio [OR]:1.43,95%confidence interval [CI]:1.02-2.06,P =0.035),donor terminal serum creatinine (OR:1.27,95%C7:1.06-1.62,P =0.023),warm ischemic time (OR:3.45,95%CI:1.97-6.37,P =0.002),and terminal resistance (OR:3.12,95%CI:1.76-6.09,P =0.012)were independent predictors of DGF.Cox proportional hazards analysis showed that terminal resistance (hazard ratio:2.06,95%C1:1.32-5.16,P =0.032)significantly affected graft survival. Conclusion:Increased HMP pressure improves graft perfusion but does not affect DGF incidence or 1-year graft survival.展开更多
Background: Hypothermic machine perfusion(HMP) is being used more often in cardiac death kidney transplantation; however, the significance of assessing organ quality and predicting delayed graft function(DGF) by HMP p...Background: Hypothermic machine perfusion(HMP) is being used more often in cardiac death kidney transplantation; however, the significance of assessing organ quality and predicting delayed graft function(DGF) by HMP parameters is still controversial. Therefore,we used a readily available HMP variable to design a scoring model that can identify the highest risk of DGF and provide the guidance and advice for organ allocation and DCD kidney assessment.Methods: From September 1, 2012 to August 31, 2016, 366 qualified kidneys were randomly assigned to the development and validation cohorts in a 2:1 distribution. The HMP variables of the development cohort served as candidate univariate predictors for DGF. The independent predictors of DGF were identified by multivariate logistic regression analysis with a P < 0.05. According to the odds ratios(ORs) value, each HMP variable was assigned a weighted integer, and the sum of the integers indicated the total risk score for each kidney. The validation cohort was used to verify the accuracy and reliability of the scoring model.Results: HMP duration(OR = 1.165, 95% confidence interval [CI ]: 1.008–1.360, P = 0.043), resistance(OR = 2.190, 95%CI: 1.032–10.20, P < 0.001), and flow rate(OR = 0.931, 95% CI: 0.894–0.967, P = 0.011) were the independent predictors of identified DGF. The HMP predictive score ranged from 0 to 14, and there was a clear increase in the incidence of DGF, from the low predictive score group to the very high predictive score group. We formed four increasingly serious risk categories(scores 0–3, 4–7, 8–11, and 12–14)according to the frequency associated with the different risk scores of DGF. The HMP predictive score indicates good discriminative power with a c?statistic of 0.706 in the validation cohort, and it had significantly better prediction value for DGF compared to both terminal flow(P = 0.012) and resistance(P = 0.006).Conclusion: The HMP predictive score is a good noninvasive tool for assessing the quality of DCD kidneys, and it is potentially useful for physicians in making optimal decisions about the organs donated.展开更多
Background:Delayed graft function(DGF)is the main cause of renal function failure after kidney transplantation.This study aims at investigating the value of hypothermic machine perfusion(HMP)parameters combined with p...Background:Delayed graft function(DGF)is the main cause of renal function failure after kidney transplantation.This study aims at investigating the value of hypothermic machine perfusion(HMP)parameters combined with perfusate biomarkers on predicting DGF and the time of renal function recovery after deceased donor(DD)kidney transplantation.Methods:HMP parameters,perfusate biomarkers and baseline characteristics of 113 DD kidney transplantations from January 1,2019 to August 31,2019 in the First Affiliated Hospital of Xi’an Jiaotong University were retrospectively analyzed using univariate and multivariate logistic regression analysis.Results:In this study,the DGF incidence was 17.7%(20/113);The multivariate logistic regression results showed that terminal resistance(OR:1.879,95%CI 1.145-3.56)and glutathione S-transferase(GST)(OR=1.62,95%CI 1.23-2.46)were risk factors for DGF;The Cox model analysis indicated that terminal resistance was an independent hazard factor for renal function recovery time(HR=0.823,95%CI 0.735-0.981).The model combining terminal resistance and GST(AUC=0.888,95%CI:0.842-0.933)significantly improved the DGF predictability compared with the use of terminal resistance(AUC=0.756,95%CI 0.693-0.818)or GST alone(AUC=0.729,95%CI 0.591-0.806).Conclusion:According to the factors analyzed in this study,the combination of HMP parameters and perfusate biomarkers displays a potent DGF predictive value.展开更多
Background Large animal cardiopulmonary bypass （CPB） models are expensive,and prevent assessment of neurocognitive function,and difficulties with long-term recovery.The purpose of this study was to establish a novel...Background Large animal cardiopulmonary bypass （CPB） models are expensive,and prevent assessment of neurocognitive function,and difficulties with long-term recovery.The purpose of this study was to establish a novel rat model of cardiopulmonary bypass for deep hypothermic circulatory arrest without blood priming.Methods Twenty adult male Sprague-Dawley rats weighing 450-560 g were randomized to CPB with deep hypothermic circulatory arrest （DHCA） and control groups,with 10 rats each.The experimental protocols,including blood and crystalloid fluid administration,anesthesia,orotracheal intubation,ventilation,cannulation,and heparinization were identical in both groups.After inducing cardiac arrest,the circuit was turned off and rats were left in a DHCA state for 15 minutes.Rats were rewarmed to 34℃ to 35℃ over a period of 36 to 42 minutes using CPB-assisted rewarming,a heating blanket,and a heating lamp along with administration of 0.1 mEq of sodium bicarbonate and 0.14 mEq of calcium chloride.The remaining priming volume was reinfused and animals were weaned from CPB.Results All CPB with DHCA processes were successfully achieved.Blood gas analysis and hemodynamic parameters were in the normal range.The vital signs of all rats were stable.Conclusions Our CPB circuit has several novel features,including a small priming volume,active cooling/rewarming processes,vacuum-assisted venous drainage,peripheral cannulation without thoracotomy or stemotomy,and an accurate means of monitoring peripheral tissue oxygenation.展开更多
Background Organ preservation keeps the quality of the organs under prolonged ischemia. Continuous machineperfusions are gaining an important position in clinical research and practice. The aim of this study was to ev...Background Organ preservation keeps the quality of the organs under prolonged ischemia. Continuous machineperfusions are gaining an important position in clinical research and practice. The aim of this study was to evaluate theprotective effect of continuous hypothermic machine perfusion transport system （AirdriveTM） on cold ischemic injury ofcanine kidney.展开更多
The chronic shortage of organs and tissues for transplantation represents a dramatic burden on healthcare systems worldwide.Tissue engineering offers a potential solution to address these shortages,but several challen...The chronic shortage of organs and tissues for transplantation represents a dramatic burden on healthcare systems worldwide.Tissue engineering offers a potential solution to address these shortages,but several challenges remain,with prevascularization being a critical factor for in vivo survival and integration of tissue engineering products.Concurrently,a different challenge hindering the clinical implementation of such products,regards their efficient preservation from the fabrication site to the bedside.Hypothermia has emerged as a potential solution for this issue due to its milder effects on biologic systems in comparison with other cold preservation methodologies.Its impact on prevascularization,however,has not been well studied.In this work,3D prevascularized constructs were fabricated using adipose-derived stromal vascular fraction cells and preserved at 4◦C using Hypothermosol or basal culture media(α-MEM).Hypothermosol efficiently preserved the structural and cellular integrity of prevascular networks as compared to constructs before preservation.In contrast,the use ofα-MEM led to a clear reduction in prevascular structures,with concurrent induction of high levels of apoptosis and autophagy at the cellular level.In vivo evaluation using a chorioallantoic membrane model demonstrated that,in opposition toα-MEM,Hypothermosol preservation retained the angiogenic potential of constructs before preservation by recruiting a similar number of blood vessels from the host and presenting similar integration with host tissue.These results emphasize the need of studying the impact of preservation techniques on key properties of tissue engineering constructs such as prevascularization,in order to validate and streamline their clinical application.展开更多
We read with great interest the study of Mergental et al.(1)reporting the 5-year outcomes of the VITTAL trial(2,3).VITTAL was a prospective,non-randomized,single-arm trial that tested end-ischemic oxygenated normother...We read with great interest the study of Mergental et al.(1)reporting the 5-year outcomes of the VITTAL trial(2,3).VITTAL was a prospective,non-randomized,single-arm trial that tested end-ischemic oxygenated normothermic machine perfusion(NMP)with a“back-to-base”strategy to evaluate,and potentially transplant,liver grafts declined by all liver transplantation(LT)centers in the United Kingdom.For a liver to be considered viable,it had to metabolize perfusate lactate to a concentration of≤2.5 mmol/L within four hours from the start of perfusion,and meet at least two of the following criteria:bile production without a defined quantity;maintenance of perfusate pH above 7.3;glucose consumption in the perfusate;maintenance of stable arterial and portal flow above 150 and 500 mL/min,respectively;maintenance of graft suppleness and homogeneous perfusion(4).Thirty-one discarded human donor livers underwent viability testing by using end-ischemic NMP,of which 22(71%)livers were subsequently transplanted.The primary outcome of the trial was graft survival rate at 90 days and it was 100%.展开更多
基金funded by the National Natural Science Foundation of China(No.81971759 and No.82171604)the Guangdong Basic and Applied Basic Research Foundation(2023B1515020108)+3 种基金the Science and Technology Program of Guangzhou(202206010089)the Excellent Talents Training Project of The Sixth Affiliated Hospital of Sun Yat-sen University(R20210217202601970)the Postdoctoral Fellowship Program of CPSF(GZC20233216)the Basic and Applied Basic Research Foundation of Guangdong Province(2021A1515111195).
文摘The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid(5-ALA).Furthermore,this study aimed to explore the underlying mechanism of the protective effects of 5-ALA.First,we collected and stored mouse testicular fragments in different media,including Hank’s balanced salt solution(HBSS;n=5),Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F12;n=5),and alpha-minimum essential medium(αMEM;n=5).Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group(P<0.05)and theαMEM group(P<0.01).Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA(0[control],1 mmol l−1,2 mmol l−1,and 5 mmol l−1)to determine the most effective concentration of 5-ALA.The 2 mmol l−15-ALA group(n=3)presented the highest positive rate of spermatogonial stem cells compared with those in the control,1 mmol l−1,and 5 mmol l−15-ALA groups.Finally,the tissue fragments were preserved in HBSS with control(n=3)and 2 mmol l−15-ALA(n=3)under low-temperature conditions.A comparative analysis was performed against fresh testes(n=3)to elucidate the underlying mechanism of 5-ALA.Gene set enrichment analysis(GSEA)for WikiPathways revealed that the p38 mitogen-activated protein kinase(MAPK)signaling pathway was downregulated in the 2 mmol l−15-ALA group compared with that in the control group(normalized enrichment score[NES]=−1.57,false discovery rate[FDR]=0.229,and P=0.019).In conclusion,these data suggest that using 2 mmol l−15-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
基金supported by grants from 2020 Guangdong Provincial and Municipal Joint Fund For Basic and Applied Basic Research Project(2020B1515120031)2020 Foshan Municipal Deng Fang Distinguished Research Program(2020A007)+2 种基金Key Scientific Research Program for the development of Ex vivo Liver Perfusion System of Foshan City,China[(2019)No.42]Science and Technol-ogy Program of Guangzhou(202002030201)Guangzhou Sci-ence and Technology Plan(2025A04J3674)。
文摘Background:The use of grafts from donation after circulatory death(DCD)overcomes the inadequate donor organ supply.Our team developed a transportable dual hypothermic oxygenated machine perfusion(DHOPE)device,which initiates DHOPE at a recipient center to reduce static cold storage(SCS)time and the risk of graft failure in DCD liver transplantation.Methods:Six porcine livers per group with 30 min of warm ischemia exposure were preserved via SCS or DHOPE for 6 h and then reperfused for 12 h with whole blood to mimic transplantation.Hepatocellular and biliary function and injury were assessed in perfusate and bile samples.Molecular biomarkers and histology were compared between groups.Results:Reperfusion portal vein pressure,in a flow-constant manner,and alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP)and gamma-glutamyltransferase(γ-GGT)release were significantly lower in the DHOPE group than in the SCS group at all time points.Higher bile production paralleled the lower levels of ALP andγ-GGT in the DHOPE group.The DHOPE group secreted more total bilirubin(TBIL)in bile,resulting in decreased TBIL in the perfusate,and livers preserved with DHOPE exhibited better cholangiocellular function.Furthermore,improvements in hypoxia,the inflammatory response,cell-free microRNAs and energy metabolism were observed in the DHOPE group.There were fewer apoptotic cells and TGF-β1-positive cells in the liver parenchyma and extrahepatic bile duct in the DHOPE group than in the SCS group.Conclusions:This study demonstrates the efficacy of local 4℃DHOPE to protect porcine liver grafts from30-min warm ischemia damage.
基金Supported by National Science and Technology Major Project,No.2012ZX10002-017Natural Science Foundation of China for Innovative Research Group,No.81121002+4 种基金National Natural Science Foundation of China,No.81000137 and No.81470891The Qianjiang Talent Program of Zhejiang Province,China,No.2012R10045the Scientific Research Program for the Returned Overseas Chinese Scholars,Ministry of Health,China,No.J20112008National High Technology Research and Development Program of China for Young Scientists(863 Program),No.2015AA020923Ministry of Education,Zhejiang Province,China,No.Y201328095
文摘AIM: To optimize the perfusates used for hypothermicmachine perfusion(HMP).METHODS: Sprague-Dawley rats were assigned randomly to three groups(n = 12 per group) that received either saline, University of Wisconsin coldstorage solution(UW) or histidine-tryptophan-ketoglutarate solution(HTK) as the perfusate. Each group was divided into two subgroups: static cold storage(SCS) and HMP(n = 6 per subgroup). The liver graft was retrieved according to the method described by Kamada. For the SCS group, the graft was directly placed into cold perfusate(0-4?℃) for 6 h after liver isolation while the portal vein of the graft was connected to the perfusion machine for the HMP group. Then the perfusates were collected at different time points for analysis of aspartate aminotransferase(AST), alanine transaminase(ALT) and lactate dehydrogenase(LDH) levels. Liver tissues were obtained for evaluation of histology, dry/wet weight(D/W) ratio, and malondialdehyde(MDA) and adenosine-triphosphate(ATP) levels. The portal vein pressure and velocity were monitored in real time in all HMP subgroups.RESULTS: Comparison of HMP and SCS: Regardless of the perfusate, HMP improved the architecture of donor graft in reducing the congestion around sinusoids and central vein and maintaining sinusoid lining in morphology; HMP improved liver function in terms of ALT, AST and LDH, especially during the 3-6 h period(SCS vs HMP using saline: ALT3, 225.00 ± 105.62 vs 49.50 ± 18.50, P = 0.047; LDH3, 1362.17 ± 563.30 vs 325.75 ± 147.43, P = 0.041; UW: LDH6, 2880.14 ± 948.46 vs 2135.00 ± 174.27, P = 0.049; HTK, AST6, 307.50 ± 52.95 vs 185.20 ± 20.46, P = 0.041); HMP decreased MDA level(saline, 2.79 ± 0.30 vs 1.09 ± 0.09, P = 0.008; UW, 3.01 ± 0.77 vs 1.23 ± 0.68, P = 0.005; HTK, 3.30 ± 0.52 vs 1.56 ± 0.22, P = 0.006). Comparison among HMP subgroups: HTK showed less portal vein resistance than UW and saline(vs saline, 3.41 ± 0.49 vs 5.00 ± 0.38, P < 0.001; vs UW, 3.41 ± 0.49 vs 4.52 ± 0.63, P = 0.007); UW reduced edema most efficiently(vs saline, 0.68 ± 0.02 vs 0.79 ± 0.05, P = 0.013), while HTK maintained ATP levels best(vs saline, 622.60 ± 29.11 vs 327.43 ± 44.66, P < 0.001; vs UW, 622.60 ± 29.11 vs 301.80 ± 37.68, P < 0.001).CONCLUSION: HMP is superior to SCS in maintaining both architecture and function of liver grafts. Further, HTK was found to be the optimal perfusate for HMP.
基金Project supported by the National Natural Science Foundation of China(No.81871541)。
文摘Oxidative stress and apoptosis are the key factors that limit the hypothermic preservation time of donor hearts to within 4–6 h.The aim of this study was to investigate whether the histone deacetylase 3(HDAC3)inhibitor RmGodFyPn9 a6 m6 i cc opualrda mpreotteercst daugrianing srt ecpaerrdfiuasci oinn juwreyr ei nedvuacleuda tebyd.pTrholeo nexgperde shsyipoont haenrdm pich opsrpehsoerryvlaattiioonn.leRvaet lsh eoaf rtms awmemrea lihayn-??STE20-like kinase-1(Mst1)and Yes-associated protein(YAP)were determined by western blotting.Cell apoptosis was measured by the terminal deoxynucleotidyl-transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)method.Addition of RGFP966 in Celsior solution significantly inhibited cardiac dysfunction induced by hypothermic preservation.RGFP966 inhibited the hypothermic preservation-induced increase of the phosphorylated(p)-Mst1/Mst1 and p-YAP/YAP ratios,prevented a reduction in total YAP protein expression,and increased the nuclear YAP protein level.Verteporfin(VP),a small molecular inhibitor of YAP–transcriptional enhanced associate domain(TEAD)interaction,partially abolished the protective effect of RGFP966 on cardiac function,and reduced lactate dehydrogenase activity and malondialdehyde content.RGFP966 increased superoxide dismutase,catalase,and glutathione peroxidase gene and protein expression,which was abolished by VP.RGFP966 inhibited hypothermic preservation-induced overexpression of B-cell lymphoma protein 2(Bcl-2)-associated X(Bax)and cleaved caspase-3,increased Bcl-2 mRNA and protein expression,and reduced cardiomyocyte apoptosis.The antioxidant and anti-apoptotic effects of RGFP966 were cancelled by VP.The results suggest that supplementation of Celsior solution with RGFP966 attenuated prolonged hypothermic preservation-induced cardiac dysfunction.The mechanism may involve inhibition of oxidative stress and apoptosis via inactivation of the YAP pathway.
基金supported by the National Natural Science Foundation of China,No.81401084(to XHW)the Beijing Municipal Administration of Hospital Ascent Plan in China,No.DFL20150802(to TLW)+2 种基金the Beijing 215 High Level Healthcare Talent Plan Academic Leader in China,No.008-0027(to TLW)the Beijing Municipal Commission of Health and Family Planning in China,No.PXM2017_026283_000002(to TLW)the Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support in China,No.ZYLX201706(to TLW),303-01-005-0137-11(to TLW),65683.00(to TLW)
文摘Using deep hypothermic circulatory arrest, thoracic aorta diseases and complex heart diseases can be subjected to corrective procedures. However, mechanisms underlying brain protection during deep hypothermic circulatory arrest are unclear. After piglet models underwent 60 minutes of deep hypothermic circulatory arrest at 14°C, expression of microRNAs(miRNAs) was analyzed in the hippocampus by microarray. Subsequently, TargetScan 6.2, RNA22 v2.0, miRWalk 2.0, and miRanda were used to predict potential targets, and gene ontology enrichment analysis was carried out to identify functional pathways involved. Quantitative reverse transcription-polymerase chain reaction was conducted to verify miRNA changes. Deep hypothermic circulatory arrest altered the expression of 35 miRNAs. Twenty-two miRNAs were significantly downregulated and thirteen miRNAs were significantly upregulated in the hippocampus after deep hypothermic circulatory arrest. Six out of eight targets among the differentially expressed miRNAs were enriched for neuronal projection(cyclin dependent kinase, CDK16 and SLC1 A2), central nervous system development(FOXO3, TYRO3, and SLC1 A2), ion transmembrane transporter activity(ATP2 B2 and SLC1 A2), and interleukin-6 receptor binding(IL6 R)– these are the key functional pathways involved in cerebral protection during deep hypothermic circulatory arrest. Quantitative reverse transcription-polymerase chain reaction confirmed the results of microarray analysis. Our experimental results illustrate a new role for transcriptional regulation in deep hypothermic circulatory arrest, and provide significant insight for the development of miRNAs to treat brain injuries. All procedures were approved by the Animal Care Committee of Xuanwu Hospital, Capital Medical University, China on March 1, 2017(approval No. XW-INI-AD2017-0112).
基金Supported by the National Natural Science Foundation,No.81470896the Project of Development and Innovation Team of Ministry of Education,No.IRT_16R57
文摘AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex vivo by hypothermic machine perfusion(HMP).METHODS Eighteen young(4 mo old) and 18 aged(17 mo old)healthy male SD rats were selected and randomly divided into three groups: control group, UW solution perfusion group(UWP), and UW solution with metformin perfusion group(MUWP). Aspartate aminotransferase(AST), alanine aminotransferase(ALT), lactate dehydrogenase(LDH), interleukin-18(IL-18), and tumor necrosis factor-alpha(TNF-α) in the perfused liquid were tested. The expression levels of AMPK and endothelial nitric oxide synthase(e NOS) in liver sinusoidal endothelial cells were also examined.Additionally, microscopic evaluation of the harvested perfused liver tissue samples was done. RESULTS AST, ALT, LDH, IL-18 and TNF-α levels in the young and aged liver-perfused liquid were, respectively,significantly lower in the MUWP group than in the UWP group(P < 0.05), but no significant differences were found between the young and aged MUWP groups.Metformin increased the expression of AMPK and e NOS protein levels, and promoted the extracellular release of nitric oxide through activation of the AMPK-e NOS mediated pathway. Histological examination revealed that in the MUWP group, the extent of liver cells and tissue damage was significantly reduced compared with the UWP group.CONCLUSION The addition of metformin to the UW preservative solution for ex vivo HMP can reduce rat liver injury during cold ischemia, with significant protective effects on livers, especially of aged rats.
文摘The technique for bloodless hepatic resection using the total hepatic vascular isolation under the normothermic or hypothermic perfusion was reported to deal with the large liver tumor involving in the liver hilum,the main hepatic veins or the retrohepatic vena cava.The original Heaney's and Fortner's methods were modified so that the technique could be simpler and more practicable to perform otherwise hazardous liver resection.During the past 4 year,major hepatic resection with the normothermic or hypothermic total vascular exclusion technique was successfully performed on 19 patients with liver tumors in our department.Among the 19 cases,16 underwent hepatic resection with the normothermic selective total vascular exclusion(extended right lobectomy in 5 cases,extended left lobectomy in 3 cases;right lobectomy in 5 cases;central segmentectomy in 3 cases)and 3 with the total vascular isolation and in situ cold perfusion(extended left lobectomy in 2 case,extended right lobectong in 1case).We believe that the technique of normothermic vascular exclusion may be indicated to deal with the lesion close to the hepatic veins and the retrohepatic vena cava.However,for more complicated hepatic resection,the hypothermic perfusion technique should be considered to prolong the safety of ischemic tune of the liver.The preliminary experience in the clinical application using the above technique is reported.
基金Technology Innovation and Application Development Project of Chongqing,No.2021yc-cxfz30016.
文摘BACKGROUND Persistent left superior vena cava(PLSVC),a relatively rare thoracic vascular malformation,can inconvenience perfusionists and operators when encountered during deep hypothermic circulatory arrest(DHCA).CASE SUMMARY Herein,we describe the case of a patient with concurrent giant aortic arch aneurysm,aortic stenosis,and PLSVC.To treat these conditions,we performed right hemiarch and aortic valve replacements under DHCA.Notably,we applied“bilateral superior vena cava retrograde cerebral perfusion(RCP)”for cerebral protection,which significantly optimized the surgical procedure and reduced the risk of postoperative complications.The patient was discharged 14 d after surgery with no complications.CONCLUSION Surgical intervention for PLSVC under DHCA can be performed using the bilateral superior vena cava RCP approach.
基金This study was reviewed and approved by our local Research Ethics Board(Bio-REB#1894).
文摘BACKGROUND Hypothermic machine perfusion(HMP)has demonstrated benefits in terms of early kidney transplant function compared to static cold storage.While longer preservation times have shown detrimental effects,a previous paired study indicated that longer pump times(the second kidney in a pair)might lead to improved outcomes.AIM To revisit the prior paired study's somewhat unexpected results by reviewing our program's experience.METHODS A total of 61 pairs of transplant recipients who received kidneys from the same donor(2012-2021)were analyzed.Patients were divided into two groups depending on whether they were transplanted first(K1)or second(K2).Therefore,the patients in each pair had identical donor characteristics,except for time on the pump.Statistical analyses included Kaplan-Meyer analysis and paired tests,including McNemar's test,student's paired t-test,or Wilcoxon's test,as appropriate.RESULTS The two groups of recipients had similar demographics(age,body mass index,diabetes,time on dialysis,sensit-ization and retransplants).Cold ischemic times for K1 and K2 were 8.9(95%CI:7.9,9.8)and 14.7 hours(13.7,15.8)(P<0.0001),respectively.Overall,K2 had a higher rate of freedom from biopsy-proven acute rejection at 1 year(P=0.015).Delayed graft function was less common in K2,12/61(20%)than in K1,20/61(33%)(P=0.046).Finally,K2 showed a higher graft survival than K1(P=0.023).CONCLUSION Our results agree with a previous study that suggested possible advantages to longer pump times.Both studies should encourage further research into HMP's potential anti-inflammatory effect.
文摘Background:Liver transplantation(LT)is a crucial treatment for end-stage liver disease,but the limited organ supply has led to the use of extended criteria donors(ECD).The implementation of dynamic preservation techniques like hypothermic oxygenated machine perfusion(HOPE)is crucial in improving outcomes for ECD grafts.However,graft contamination and infection are a concern.This study aimed to evaluate the risk of infections within 10 days from LT between HOPE and static cold storage(SCS)groups and postoperative complications.Methods:A retrospective cohort study was conducted,including LT recipients transplanted at a single-center from March 2016 to June 2023.Patients were divided into HOPE and SCS groups,and propensity score matching was used to select comparable cohorts.Data on patient and donor characteristics were analyzed.Results:After propensity score matching,a cohort of 370(HOPE,n=185;SCS,n=185)patients was selected for analysis.The study found no significant differences in the rate of clinically relevant infections,microbiological positive samples,or donor-derived infections within 10 days between HOPE and SCS groups.Postoperative outcomes,as well as patient and graft survival,were also similar between the two groups.The study showed that HOPE is a feasible and safe approach,with a comparable risk of infection.Conclusions:The study results indicate that HOPE use in LT does not increase the risk of infection and is associated with similar patient and graft survival outcomes compared to SCS.These findings confirm the safety and efficacy of HOPE in LT and its potential to expand the donor pool without compromising recipient outcomes.
基金funded by EU Horizon 2020 research and innovation program under the European Research Council(ERC)grant CapBed(805411)the Portuguese Science and Technology Foundation(FCT)UIDB/04539/2020,UIDP/04539/2020+1 种基金The National Mass Spectrometry Network(POCI-01–0145-FEDER-402–022125 Ref.ROTEIRO/0028/2013)SFR acknowledges doctoral fellowship PD/BD/135252/2017 and RAP acknowledges individual grant CEECIND/05623/2022.
文摘Cell sheet(CS)-based approaches hold significant potential for tissue regeneration,relying on the extracellular matrix(ECM)for success.Like in native tissues,the ECM provides structural and biochemical support for cellular homeostasis and function.Effective preservation strategies that maintain ECM integrity are critical to enhance the therapeutic potential of CS-based approaches.While cryogenic and hypothermic preservation methods offer potential solutions,their impact on CS ECM structure is not fully understood.Therefore,a comprehensive analysis of the ECM of hASCs CS following cryogenic and hypothermic preservation for 3 and 7 days,was conducted.Although proteomic analysis indicated that cryopreservation had no significant effect on the overall composition of the ECM,it induced significant ECM structural alterations,particularly disrupting collagen organization,which was not observed following hypothermic preservation.These structural changes were accompanied by alterations in mechanical properties,including a reduction in elastic modulus.In contrast,hypothermic preservation maintained ECM integrity and mechanical properties similar to the control.The notable ECM structural changes following cryogenic preservation can potentially impact cellular behavior,including adhesion,proliferation,and differentiation,thereby affecting the efficacy of CS therapies in vivo.This suggests that hypothermia may offer a promising alternative to cryopreservation for preserving CS integrity and functionality.
文摘Background:Ischemia reperfusion injury(IRI)is a major contributing factor to organ damage in liver transplantation(LT)impacting donor organ quality and patient survival.IRI-inflicted graft injury can be reduced by using hypothermic oxygenated machine perfusion(HOPE)as a preservation strategy instead of static cold storage(SCS).The endothelial glycocalyx is highly sensitive to IRI and its degradation during graft preservation and reperfusion was previously associated with inferior postoperative outcome after LT.Here,we aimed to measure glycocalyx degradation during and after HOPE in order to evaluate its potential for viability-assessment during machine perfusion and outcome prediction in patients undergoing LT.Methods:Glycocalyx degradation was quantified via enzyme-linked immunoassay(ELISA)for its main component syndecan-1(Sdc-1)in serum of 40 patients undergoing LT after HOPE.In addition,Sdc-1 was evaluated at multiple time points during HOPE.Patients were followed up for 3.5 years to assess postoperative complications including morbidity,the development of early allograft dysfunction(EAD)and graft survival.Results:Liver grafts which later developed EAD showed significantly higher Sdc-1 concentrations after 60 min of HOPE compared to grafts exhibiting normal postoperative function(P=0.02).Receiver operating characteristic analysis revealed a strong predictive potential with an area under the curve of 0.73.A cut-off at 808 ng/mL Sdc-1 at 60 min of HOPE allowed identification of a high-risk group with an incidence of EAD of 66.7%.Sdc-1 concentrations increased during all types of HOPE but were significantly higher in HOPE versus dual HOPE(D-HOPE)after 120 min of perfusion(P=0.02).Conclusions:Sdc-1 evaluated at 60 min during HOPE allows prediction of EAD after LT.Accordingly,Sdc-1 should be considered a potential additional biomarker for viability assessment during HOPE.
基金grants from the Fundamental Research Funds for the Central Universities (No.xjj2018091)Major Clinical Research Projects of the First Affiliated Hospital of Xi'an Jiaotong University (No.XJTU 1AF-CRF-2015-005)+1 种基金Scientific and Technological Breakthrough in Social Development of Shaanxi Province (No.2016SF-246)National Natural Science Foundation of China (No.81670681and 81760137).
文摘Background:Vascular resistance and flow rate during hypotherrnic machine perfusion (HMP)of kidneys is correlated with graft function. We aimed to determine the effects of increasing HMP pressure versus maintaining the initial pressure on kidney transplantation outcomes. Methods:We retrospectively reviewed the data of 76 primary transplantation patients who received HMP-preserved kidneys from 48 donors after cardiac death between September 1,2013,and August 31,2015.HMP pressure was increased from 30 to 40mmHg (1mmHg =0.133kPa)in kidneys with poor flow and/or vascular resistance (increased pressure [IP]group;36 patients);otherwise,the initial pressure was maintained (constant pressure group;40 patients).Finally,the clinical characteristics and transplantation outcomes in both groups were assessed. Results:Delayed graft function (DGF)incidence,1-year allograft,patient survival,kidney function recovery time,and serum creatinine level on day 30 were similar in both groups,with improved flow and resistance in the IP group.Among patients with DGF,kidney function recovery time and DGF duration were ameliorated in the IP group.Multivariate logistic regression analysis revealed that donor hypertension (odds ratio [OR]:1.43,95%confidence interval [CI]:1.02-2.06,P =0.035),donor terminal serum creatinine (OR:1.27,95%C7:1.06-1.62,P =0.023),warm ischemic time (OR:3.45,95%CI:1.97-6.37,P =0.002),and terminal resistance (OR:3.12,95%CI:1.76-6.09,P =0.012)were independent predictors of DGF.Cox proportional hazards analysis showed that terminal resistance (hazard ratio:2.06,95%C1:1.32-5.16,P =0.032)significantly affected graft survival. Conclusion:Increased HMP pressure improves graft perfusion but does not affect DGF incidence or 1-year graft survival.
基金grants from the Fundamental Research Funds for the Central Universities (No.xjj2018091)Major Clinical Research Projects of the First Affiliated Hospital of Xi'an Jiaotong University (No.XJTU1 AF-CRF-2015-005)+1 种基金Scientific and Technological Breakthrough in Social Development of Shaanxi Province (No.2016SF-246) National Natural Science Foundation of China (No.81670681 and 81760137).
文摘Background: Hypothermic machine perfusion(HMP) is being used more often in cardiac death kidney transplantation; however, the significance of assessing organ quality and predicting delayed graft function(DGF) by HMP parameters is still controversial. Therefore,we used a readily available HMP variable to design a scoring model that can identify the highest risk of DGF and provide the guidance and advice for organ allocation and DCD kidney assessment.Methods: From September 1, 2012 to August 31, 2016, 366 qualified kidneys were randomly assigned to the development and validation cohorts in a 2:1 distribution. The HMP variables of the development cohort served as candidate univariate predictors for DGF. The independent predictors of DGF were identified by multivariate logistic regression analysis with a P < 0.05. According to the odds ratios(ORs) value, each HMP variable was assigned a weighted integer, and the sum of the integers indicated the total risk score for each kidney. The validation cohort was used to verify the accuracy and reliability of the scoring model.Results: HMP duration(OR = 1.165, 95% confidence interval [CI ]: 1.008–1.360, P = 0.043), resistance(OR = 2.190, 95%CI: 1.032–10.20, P < 0.001), and flow rate(OR = 0.931, 95% CI: 0.894–0.967, P = 0.011) were the independent predictors of identified DGF. The HMP predictive score ranged from 0 to 14, and there was a clear increase in the incidence of DGF, from the low predictive score group to the very high predictive score group. We formed four increasingly serious risk categories(scores 0–3, 4–7, 8–11, and 12–14)according to the frequency associated with the different risk scores of DGF. The HMP predictive score indicates good discriminative power with a c?statistic of 0.706 in the validation cohort, and it had significantly better prediction value for DGF compared to both terminal flow(P = 0.012) and resistance(P = 0.006).Conclusion: The HMP predictive score is a good noninvasive tool for assessing the quality of DCD kidneys, and it is potentially useful for physicians in making optimal decisions about the organs donated.
基金This work was supported by the grants from the National Natural Science Foundation of China(Nos.81670681,81760137,81870514,81970668 and 81970670)the Fundamental Research Funds for the Central Universities(No.xjj2018091)+1 种基金the Clinical Research Award of the First Affiliated Hospital of Xi’an Jiaotong University(No.XJTU1AF-CRF-2019-008)the Special Supportive Program for Organ Transplantation by COTDF(No.2019JYJH04).
文摘Background:Delayed graft function(DGF)is the main cause of renal function failure after kidney transplantation.This study aims at investigating the value of hypothermic machine perfusion(HMP)parameters combined with perfusate biomarkers on predicting DGF and the time of renal function recovery after deceased donor(DD)kidney transplantation.Methods:HMP parameters,perfusate biomarkers and baseline characteristics of 113 DD kidney transplantations from January 1,2019 to August 31,2019 in the First Affiliated Hospital of Xi’an Jiaotong University were retrospectively analyzed using univariate and multivariate logistic regression analysis.Results:In this study,the DGF incidence was 17.7%(20/113);The multivariate logistic regression results showed that terminal resistance(OR:1.879,95%CI 1.145-3.56)and glutathione S-transferase(GST)(OR=1.62,95%CI 1.23-2.46)were risk factors for DGF;The Cox model analysis indicated that terminal resistance was an independent hazard factor for renal function recovery time(HR=0.823,95%CI 0.735-0.981).The model combining terminal resistance and GST(AUC=0.888,95%CI:0.842-0.933)significantly improved the DGF predictability compared with the use of terminal resistance(AUC=0.756,95%CI 0.693-0.818)or GST alone(AUC=0.729,95%CI 0.591-0.806).Conclusion:According to the factors analyzed in this study,the combination of HMP parameters and perfusate biomarkers displays a potent DGF predictive value.
基金This study was supported by grants from the National Natural Science Foundation of China （No.81371443）,Beijing Natural Science Foundation （No.7142137,No.7122056 and No.7142049）,Basic and Clinical Cooperation Project of Capital Medical University （No.13JL26）.
文摘Background Large animal cardiopulmonary bypass （CPB） models are expensive,and prevent assessment of neurocognitive function,and difficulties with long-term recovery.The purpose of this study was to establish a novel rat model of cardiopulmonary bypass for deep hypothermic circulatory arrest without blood priming.Methods Twenty adult male Sprague-Dawley rats weighing 450-560 g were randomized to CPB with deep hypothermic circulatory arrest （DHCA） and control groups,with 10 rats each.The experimental protocols,including blood and crystalloid fluid administration,anesthesia,orotracheal intubation,ventilation,cannulation,and heparinization were identical in both groups.After inducing cardiac arrest,the circuit was turned off and rats were left in a DHCA state for 15 minutes.Rats were rewarmed to 34℃ to 35℃ over a period of 36 to 42 minutes using CPB-assisted rewarming,a heating blanket,and a heating lamp along with administration of 0.1 mEq of sodium bicarbonate and 0.14 mEq of calcium chloride.The remaining priming volume was reinfused and animals were weaned from CPB.Results All CPB with DHCA processes were successfully achieved.Blood gas analysis and hemodynamic parameters were in the normal range.The vital signs of all rats were stable.Conclusions Our CPB circuit has several novel features,including a small priming volume,active cooling/rewarming processes,vacuum-assisted venous drainage,peripheral cannulation without thoracotomy or stemotomy,and an accurate means of monitoring peripheral tissue oxygenation.
基金The study was supported by grants from the National Natural Science Foundation of China （No. 81270837） and the Beijing Natural Science Foundation （No. 7132107）.
文摘Background Organ preservation keeps the quality of the organs under prolonged ischemia. Continuous machineperfusions are gaining an important position in clinical research and practice. The aim of this study was to evaluate theprotective effect of continuous hypothermic machine perfusion transport system （AirdriveTM） on cold ischemic injury ofcanine kidney.
基金funded by EU Horizon 2020 research and innovation program under the ERC grant CapBed (805411)national funds+1 种基金trough the Portuguese Foundation for Science and Technology (FCT)project UIDB/50026/2020 and UIDP/50026/2020SFR acknowledges doctoral fellowship PD/BD/135252/2017,RPP individual grant IF/00347/2015,BSM individual grant DL 57/2016,LdS Scientific Employment Stimulus-Individual Call (CEEC Individual)2020.01541.CEECIND/CP1600/CT0024。
文摘The chronic shortage of organs and tissues for transplantation represents a dramatic burden on healthcare systems worldwide.Tissue engineering offers a potential solution to address these shortages,but several challenges remain,with prevascularization being a critical factor for in vivo survival and integration of tissue engineering products.Concurrently,a different challenge hindering the clinical implementation of such products,regards their efficient preservation from the fabrication site to the bedside.Hypothermia has emerged as a potential solution for this issue due to its milder effects on biologic systems in comparison with other cold preservation methodologies.Its impact on prevascularization,however,has not been well studied.In this work,3D prevascularized constructs were fabricated using adipose-derived stromal vascular fraction cells and preserved at 4◦C using Hypothermosol or basal culture media(α-MEM).Hypothermosol efficiently preserved the structural and cellular integrity of prevascular networks as compared to constructs before preservation.In contrast,the use ofα-MEM led to a clear reduction in prevascular structures,with concurrent induction of high levels of apoptosis and autophagy at the cellular level.In vivo evaluation using a chorioallantoic membrane model demonstrated that,in opposition toα-MEM,Hypothermosol preservation retained the angiogenic potential of constructs before preservation by recruiting a similar number of blood vessels from the host and presenting similar integration with host tissue.These results emphasize the need of studying the impact of preservation techniques on key properties of tissue engineering constructs such as prevascularization,in order to validate and streamline their clinical application.
文摘We read with great interest the study of Mergental et al.(1)reporting the 5-year outcomes of the VITTAL trial(2,3).VITTAL was a prospective,non-randomized,single-arm trial that tested end-ischemic oxygenated normothermic machine perfusion(NMP)with a“back-to-base”strategy to evaluate,and potentially transplant,liver grafts declined by all liver transplantation(LT)centers in the United Kingdom.For a liver to be considered viable,it had to metabolize perfusate lactate to a concentration of≤2.5 mmol/L within four hours from the start of perfusion,and meet at least two of the following criteria:bile production without a defined quantity;maintenance of perfusate pH above 7.3;glucose consumption in the perfusate;maintenance of stable arterial and portal flow above 150 and 500 mL/min,respectively;maintenance of graft suppleness and homogeneous perfusion(4).Thirty-one discarded human donor livers underwent viability testing by using end-ischemic NMP,of which 22(71%)livers were subsequently transplanted.The primary outcome of the trial was graft survival rate at 90 days and it was 100%.
