Evolution and innovation of preservation fluid for donor liver / 器官移植

Organ preservation fluid could mitigate cold ischemia injury and maintain normal function of the grafts. At present, how to reduce a series of injury caused by cold ischemia of donor liver and improve the preservation quality of grafts are the hot and challenging spots in this field. Currently, preservation fluid in clinical practice has not achieved ideal preservation effect, especially for the protection of marginal donor organs. In the context of severe donor shortage, the key solution is still to explore the optimal preservation protocol for donor liver to prevent grafts from cold ischemia injury. In this article, the mechanism of donor liver injury during cold ischemia, the classification and evolution of donor liver preservation fluid were summarized, the development direction and challenges of donor liver preservation fluid were discussed, aiming to provide novel ideas and references for the research and development of donor liver preservation fluid.

Research progress on protective effect of hydrogen molecule and its products on lung transplantation / 器官移植

Ischemia-reperfusion injury after lung transplantation is the main cause of primary graft dysfunction, which will subsequently reduce the function of lung allograft and lower the overall survival rate of lung transplant recipients. As a physiological regulatory molecule, hydrogen molecule has the functions of anti-inflammation, easing oxidative stress, alleviating direct cell injury and mitigating epithelial edema. Recent studies have demonstrated that hydrogen molecule and its products (hydrogen and hydrogen-rich solution) could significantly mitigate ischemia-reperfusion injury and postoperative complications after lung transplantation. In this article, the protective effect and exact mechanism of hydrogen molecule and its products in lung transplantation were reviewed, aiming to provide theoretical basis for the application of hydrogen molecule and its products as a novel treatment for lung transplantation-related complications, enhance the overall prognosis and improve the quality of life of lung transplant recipients

Efetividade da máquina de perfusão hipotérmica versus armazenamento estático para preservação do rim de doador falecido: revisão rápida / Effectiveness of hypothermic machine perfusion versus cold static storage for deceased donor kidney preservation: rapid review

Tecnologia: máquina de perfusão hipotérmica. Indicação: Transplante renal de doador falecido. Pergunta: Qual a efetividade da máquina de perfusão hipotérmica (HMP) para a preservação do rim de doador falecido, quando comparada ao armazenamento estático a frio (SCS)? Objetivo. Avaliar a efetividade da máquina de perfusão hipotérmica na preservação do rim de doador falecido, em comparação com o armazenamento estático a frio. Métodos: Revisão de revisões sistemáticas (overview) do tipo revisão rápida. Foi realizado um levantamento bibliográfico nas bases de dados: PubMed, Embase, BVS, Epistemonikos, Cochrane Library e em bases de registro de protocolos de revisões sistemáticas e ensaios clínicos, utilizando descritores e estratégias de busca predefinidas. A avaliação da qualidade metodológica dos estudos incluídos foi feita através da ferramenta AMSTAR-2 (Assessing the Methodological Quality of Systematic Reviews Version 2). Resultados: Duas revisões sistemáticas atenderam aos critérios de elegibilidade e foram incluídas na análise. Uma delas apresentou alto nível de qualidade metodológica. Conclusão: O uso da HMP para a preservação de rins de doadores falecidos foi associado a melhores desfechos clínicos relacionados à função retardada e à sobrevida do enxerto e foi considerado custo-efetivo, quando comparado ao SCS. Faz-se necessária a geração de evidências mais robustas acerca dos custos e benefícios do uso desta tecnologia no âmbito do SUS

Technology: hypothermic machine perfusion. Indication: Deceased donor kidney transplantation. Question: How effective is hypothermic machine perfusion (HMP) for preserving deceased donor kidneys compared to static cold storage (SCS)? Objective: To evaluate the effectiveness of the hypothermic machine perfusion in preserving the deceased donor kidney, compared to static cold storage. Methods: Rapid review of systematic reviews (overview). A bibliographic survey was carried out in the databases: PubMed, Embase, VHL, Epistemonikos, Cochrane Library and in databases of systematic review protocols and clinical trials, using predefined descriptors and search strategies. The assessment of the methodological quality of the included studies was performed using the AMSTAR-2 tool (Assessing the Methodological Quality of Systematic Reviews Version 2). Results: Two systematic reviews met the eligibility criteria and were included in the analysis. One of them performed a high level of methodological quality. Conclusion: The use of HMP for the preservation of deceased donor kidneys was associated with better clinical outcomes related to delayed graft function and graft survival and was considered cost-effective. It is necessary to generate more evidence about the costs and benefits of using this technology within the Brazilian Unified System of Healthcare (SUS)

A modified mouse model of orthotopic left lung transplantation based on "pendulum" anastomosis of the reverse-view plane (with video demonstration) / 器官移植

Objective To modify the mouse model of orthotopic left lung transplantation from different perspectives, aiming to establish a simpler, faster and stabler mouse model of lung transplantation. Methods Based on preliminary modified rat model of orthotopic left lung transplantation established by our team, varying extent of modifications were made regarding the tracheal intubation, cannula preparation and anastomosis procedures of orthotopic left lung transplantation in the recipient mice. Orthotopic left lung transplantation in 40 mice were performed by an operator with microsurgical experience. The dissection of the recipient's hilar structure was carried out at the plane of the hilar clamp model within the reverse-view, and the three branches (left main bronchus, pulmonary artery and pulmonary vein) of the pulmonary hilum were anastomosed in turn by the "pendulum" anastomosis method. The operation time of each procedure was recorded. The recipient mice were sacrificed at postoperative 2 weeks, and the incidence of postoperative complications was recorded. Results Lung transplantation was successfully completed in 40 mice, with no bronchial and vascular tearing or twisting, and no bleeding at the anastomosis site. The overall cardiopulmonary procurement time was (10.7±1.5) min, cannula preparation time was (16.2±1.5) min, cold ischemia time was (25.1±2.4) min, warm ischemia time was (19.4±1.6) min, and the total operation time was (57.2±2.9) min, respectively. During the follow-up from 6 to 14 days after surgery, one recipient mouse died of pleural effusion, probably caused by infection. No pneumothorax, thrombosis or atelectasis was found in the remaining recipient mice during postoperative follow-up. Conclusions The modified mouse model of orthotopic left lung transplantation based on "pendulum" anastomosis of the reverse-view plane possesses multiple advantages of short operation time, high success rate and few complications, which is expected to become an alternative model of studying pathological changes after lung transplantation and worthy of further application.

Clinical application of split liver transplantation technique / 器官移植

Currently, multiple difficulties exist in clinical liver transplantation, such as shortage of donor liver, increasing quantity of patients waiting for liver transplantation and lack of matching donors, etc. Some children and adult patients have little chance of undergoing liver transplantation, which also limits the development of liver transplantation. In this context, split liver transplantation emerges, in which 1 donor liver can be applied to 2 or even more recipients. It may effectively increase the utilization rate of donor liver and alleviate the shortage of donor liver. With the development of split liver transplantation, the survival rate of split liver transplantation is comparable to that of total liver transplantation. Multiple transplantation centers have routinely adopted split liver transplantation. In this article, the development of split liver transplantation, the selection and matching of donors and recipients, the split and reconstruction techniques of donor liver and postoperative complications were reviewed, aiming to provide reference for subsequent development of split liver transplantation in clinical practice and increase the chance of liver transplantation for more patients diagnosed with end-stage liver diseases.

¿Puede el Líquıdo Amnıótıco Ser una Solucıón Alternatıva de Conservacıón de Órganos para el Almacenamıento Renal en Frío? / Can Amniotic Fluid Be an Alternative Organ Preservation Solution for Cold Renal Storage?

Abstract Introduction: Kidney-transplantation is a life-saving treatment option for patients with chronic renal failure. Preserving the viability of the organ from the removal of the organ until transplantation into the recipient is one of the most essential factors affecting postransplant success. Kidney tissue is exposed to ischemia following removal of the organ from the donor, initiating some cellular events. Amniotic fluid (AF) was previously reported as a preservation solution for the liver, but not for the kidney yet. The aim of this study is to investigate the effectiveness of AF as a preserving solution for rat kidneys compared with the University of Wisconsin (UW) and Histidine-Tryptophan-Ketoglutarate (HTK), which are reported to be the most commonly used and preferred preserving solutions. Methods: Forty male Wistar albino rats were used in this study in four experimental groups. Group 1: Ringer Lactate (RL, Control) group, Group 2: HTK group, Group 3: UW group, and Group 4: AF group. A midline incision was performed, and the renal artery was isolated under ketamine and xylazine anesthesia. Solutions relevant for groups (cooled to + 4°C) were used for kidney perfusion. Nephrectomy was applied, and the removed kidneys were placed into + 4°C standard organ storage solution and stored at + 4° C for 12 hours. After 12 hours of storage, samples from the kidney tissues were fixed in 10% neutral buffered formalin. Histopathological, immunohistochemistry evaluation and apoptosis detection via TUNEL method were performed. Results: The results of the AF group were close to those of the UW and HTK groups. Tubular necrosis and vacuolization were high in the RL solution group when compared to the other ‎experimental groups. Immunohistochemistry staining for all three markers (TNF-alpha, IL-18, and iNOS) was decreased in the amniotic fluid group, similar to the UW and HTK groups. Also, the number of apoptotic cells was decreased in the AF group compared to control. Conclusions: UW, HTK, and AF had similar and higher protective effects compared to the RL solution. Thus, AF may be used as an inexpensive and readily available alternative natural tissue preservation solution.

Resumen Introducción: El trasplante de riñón es una opción de tratamiento que puede salvar la vida de pacientes con insuficiencia renal crónica. Preservar la viabilidad del órgano desde su extracción hasta el momento del trasplante en el receptor es uno de los factores principales que influyen en el éxito postrasplante. El tejido renal está expuesto a la isquemia después de la extracción del órgano del donante, lo cual da inicio a algunos eventos celulares. Existen estudios que indican que el líquido amniótico (LA) funciona como una solución de conservación para el hígado, pero aún se desconoce si sucede lo mismo con el riñón. El objetivo de este estudio es investigar la efectividad del LA como solución conservadora para los riñones de ratas, en comparación con la solución de Wisconsin (UW) y la solución de histidina-triptófano-cetoglutarato (HTK), que son los conservantes más utilizados y preferidos. Material y métodos: Se emplearon cuarenta ratas albinas macho de la cepa Wistar en este estudio, en cuatro grupos experimentales. Grupo 1: grupo solución de lactato sódico compuesta (LSC, Control); Grupo 2: grupo HTK; Grupo 3: grupo UW y Grupo 4: grupo LA. Habiendo aplicado anestesia con ketamina y xilazina, se realizó una incisión en la línea media y se aisló la arteria renal. Se utilizaron soluciones relevantes para grupos (enfriadas a + 4° C) para perfusión renal. Se realizó una nefrectomía, y los riñones extraídos fueron colocaron en una solución estándar de almacenamiento de órganos a + 4° C y se conservaron así durante 12 horas. Después de dicho periodo de almacenamiento, las muestras de los tejidos renales se fijaron en formalina tamponada neutra al 10%. Se llevaron a cabo una evaluación histopatológica e inmunohistoquímica y una detección de apoptosis mediante el método TUNEL. Resultados: Los resultados del grupo LA fueron cercanos a los de los grupos UW y HTK. La necrosis tubular y la vacuolización fueron más altas en el grupo de la solución LSC que en los otros grupos experimentales. La tinción inmunohistoquímica para los tres marcadores (TNF-alfa, IL-18 e iNOS) disminuyó en el grupo de líquido amniótico, similar a los grupos UW y HTK. Además, el número de células apoptóticas menguó en el grupo LA, en comparación con el de control. Conclusiones: UW, HTK y LA tuvieron efectos protectores similares y superiores en comparación con la solución LSC. Por lo tanto, el LA puede usarse como una solución alternativa de bajo costo para la preservación de tejidos naturales.

Comparative study between fructose 1-6 bisphosphate and histidine-tryptophan-ketoglutarate in liver preservation in rats submitted to total cold ischemia

Abstract Purpose To compare Fructose-1,6-Bisphosphate (FBP) to Histidine-Tryptophan-Ketoglutarate (HTK) in liver preservation at cold ischemia. Methods Male rats (Sprague-Dawley: 280-340g) divided into three groups (n=7): Control; Fructose-1,6-bisphosphate (FBP); Histidine-Tryptophan-Ketoglutarate (HTK). Animals underwent laparotomy-thoracotomy for perfusion of livers with saline. Livers were removed and deposited into solutions. Mitochondria were isolated to determine State 3 (S3), State 4 (S4), Respiratory Control Ratio (RCR) and Swelling (S). Liver enzymes (AST, ALT, LDH) were determined in solution. At tissue, Malondialdehyde (MDA) and Nitrate (NOx) were determined. All parameters were analyzed at 0.6 and 24 hours of hypothermic preservation. Statistics analysis were made by Mann-Whitney test (p<0.05). Results Regarding ALT, there was a difference between FBP-6h/HTK-6h, lower in HTK. Regarding AST, there was a significant difference between FBP-24h/HTK-24h, lower in FBP. Regarding NOx, there was a difference between 0h and 6h, as well as 0h and 24h for both solutions. Regarding S3, there was a significant difference in 24h compared to Control-0h for both solutions, and a significant difference between FBP-6h/FBP-24h. Regarding S4, there was a difference between Control-0h/HTK-24h and FBP-24h/HTK-24h, higher in HTK. There was a difference between Control-0h/FBP-24h for Swelling, higher in FBP. Conclusion Fructose-1,6-Bisphosphate showed better performance at nitrate and aspartate aminotransferase compared to histidine-tryptophan-ketoglutarate.

Analysis of early prognosis and risk factors of donor liver with moderate-to-severe steatosis in adult liver transplantation / 器官移植

Objective To explore the early prognosis and the risk factors of delayed graft function (DGF) of the recipients undergoing liver transplantation from donor liver with moderate-to-severe steatosis. Methods Clinical data of 475 donors and 475 recipients undergoing liver transplantation from donor liver of organ donation after citizen's death were retrospectively analyzed. According to the classification criteria of steatosis proposed by Australia National Liver Transplantation Unit (ANLTU), all recipients were divided into the S0 group (no steatosis, n=308), S1 group (mild steatosis, n=97), S2 group (moderate steatosis, n=52) and S3 group (severe steatosis, n=18), respectively. The early postoperative death and incidence of postoperative complications were statistically compared among each group. The risk factors from donors, recipients and operation leading to DGF were analyzed by univariate and multivariate logistic regression models. Results The incidence of postoperative DGF in the S2 and S3 groups was significantly higher than that in the S1 and S0 groups (all P < 0.05). The incidence of postoperative DGF in the S3 group was remarkably higher than that in the S2 group (P < 0.05). The early postoperative fatality, the incidence of primary nonfunction (PNF) of the transplant liver, postoperative bleeding, infection, biliary complications and vascular complications did not significantly differ among each group (all P > 0.05). Univariate regression analysis showed that severe steatosis of donor liver, long cold ischemia time, high model for end-stage liver disease (MELD) score and tumors of the recipients before operation were the risk factors of DGF (all P < 0.05). Multivariate logistic regression analysis demonstrated that moderate-to-severe steatosis of donor liver, cold ischemia time > 8 h and MELD score > 30 of the recipients were the independent risk factors for early postoperative DGF. Conclusions The early-stage incidence of DGF after adult liver transplantation from donor liver with moderate-to-severe steatosis is high, whereas it does not affect the early survival rate of the recipients. The selection of donor liver with moderate-to-severe steatosis should be considered in combination with cold ischemia time of the donors and MELD score of the recipients before operation, etc.

Effect of cold ischemia time on early graft function and acute rejection after liver transplantation / 器官移植

Objective To evaluate the effect of different cold ischemia time (CIT) on early graft function and acute rejection (AR) after liver transplantation. Methods Clinical data of 218 donors and recipients undergoing liver transplantation were collected and analyzed. All patients were divided into three groups according to the CIT of donor liver: group A (CIT≤6 h, n=60), group B (6 h < CIT≤10 h, n=89) and group C (CIT > 10 h, n=69). Blood samples were collected on the 1, 7 and 14 d after liver transplantation. The changes of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and adenosine triphosphate (ATP) in CD4+T cells were detected. The incidence of AR and the positive rate of C4d deposition were analyzed. Results The ALT, AST and LDH levels in each group reached the peak on the 1 d after operation, and then gradually decreased. The indexes in each group were almost equivalent on the 14 d. An interaction effect existed between postoperative time and group. After liver transplantation, ATP levels in CD4+T cells were gradually increased in each group, peaked at postoperative 7 d, and then decreased gradually. An interaction effect was noted between postoperative time and group. The incidence of AR in groups A, B and C was 10%, 12% and 28%. Compared with group C, the incidence of AR in groups A and B was decreased significantly (both P < 0.05/3). The positive rate of C4d deposition in AR recipients of groups A, B and C was 1/3, 45% and 89% respectively. Compared with group C, the positive rate of C4d deposition in group A was decreased significantly (P=0.015). Conclusions The prolongation of CIT may lead to aggravation of early-stage liver function injury after liver transplantation, which is more easily to induce humoral AR.

Máquina de perfusão versus armazenamento estático na preservação renal de doadores com morte encefálica: revisão sistemática e metanálise / Machine perfusion versus cold storage in renal preservation of deceased donors with brain death: systematic review and meta-analysis

RESUMO Com a utilização crescente da máquina de perfusão no transplante renal, tem sido constatado que a isquemia dinâmica correlaciona-se à melhora da preservação orgânica. Nesse contexto, realizamos uma revisão sistemática que procurou avaliar a eficácia do uso de máquina de perfusão portátil (LifePort Kidney Transporter Machine®), utilizada no Brasil, comparada ao armazenamento estático, no que tange à função retardada do transplante renal de doadores com morte encefálica. Foi efetuada pesquisa bibliográfica, nas bases LILACS, MEDLINE via PubMed, Scopus, Clarivate Analytics, Cochrane Library, Embase, SciELO, além de busca manual no Google acadêmico. A revisão sistemática, finalizada em abril 2017, foi constituída somente por ensaios clínicos randomizados. Para metanálise, foram avaliadas Razão de Risco e Razão de Chance. Foram identificados 86 documentos e selecionados, ao final, dois artigos com critérios de elegibilidade para metanálise, de grupos europeus e brasileiros. Nestes, 374 rins foram alocados para a máquina de perfusão, e igual número para o armazenamento estático. A função retardada do enxerto foi constatada em 84 e 110 pacientes, respectivamente. Na metanálise, foram obtidas uma Razão de Risco de 0,7568 (p=0,0151) e uma Razão de Chance de 0,6665 (p=0,0225), ambas com intervalo de confiança de 95%. A máquina de perfusão reduziu a incidência de função retardada do enxerto de doadores com morte encefálica.

ABSTRACT With the increasing use of machine perfusion in kidney transplantation, it has been observed that dynamic ischemia correlates with the improvement of organ preservation. In this context, we performed a systematic review that aimed to evaluate the efficacy of the portable machine perfusion (LifePort Kidney Transporter Machine®), used in Brazil, compared to cold storage, regarding the delayed graft function of deceased donors with brain death. Literature search was carried out in LILACS, MEDLINE via PubMed, Scopus, Clarivate Analytics, Cochrane Library, Embase, and SciELO, as well as in Google Scholar manually. The systematic review consisted only of randomized clinical trials. For meta-analysis, relative risk and odds ratio were evaluated. Eighty-six documents were identified and two papers from European and Brazilian groups were selected at the end, with eligibility criteria for meta-analysis. In these, 374 kidneys were assigned to machine perfusion and 374 kidneys were assigned to cold storage. Delayed graft function was observed in 84 and 110 patients, respectively. In meta-analysis, a risk ratio of 0.7568 (p=0.0151) and an odds ratio of 0.6665 (p=0.0225) were obtained, both with a 95% confidence interval. Machine perfusion reduced the incidence of delayed graft function of deceased donors with brain death.

Analysis of survival and influencing factors of patients with recurrent and de novo nephritis of renal allograft / 器官移植

Objective To analyze the survival and influencing factors of patients with recurrent and de novo nephritis of the renal allograft. Methods Clinical data of 95 patients undergoing pathological puncture (biopsy) of the renal allograft were retrospectively analyzed. According to the biopsy results, all recipients were assigned into the recurrent group (n=28), de novo group(n=33) and non-nephritis group (n=34). The 1-, 3- and 5-year survival was statistically analyzed and the survival rates were calculated in three groups. Kaplan-Meier survival curve was adopted to analyze the 5-year survival. Clinical data of patients with recurrent and de novo nephritis were analyzed by univariate analysis. Logistic regression analysis was utilized to analyze the influencing factors of clinical prognosis of patients with recurrent and de novo nephritis. Results The 1-year survival rate did not significantly differ among three groups (all P > 0.05). The 3-year survival rates in the de novo group and non-nephritis group were 97% and 100%, significantly higher than 86% in the recurrent group (both P < 0.05). The 5-year survival rates in the de novo group and non-nephritis group were 82% and 91%, considerably higher than 61% in the recurrent group (both P < 0.05). Logistic regression analysis demonstrated that the survival rate of patients with recurrent renal nephritis was significantly correlated with the times of renal transplantation, cold ischemia time (≥12 h), immunosuppressive regime, recovery time of postoperative serum creatinine (Scr) (≥14 d), complications at postoperative 1 month (acute renal tubular necrosis, ultra-acute rejection and acute rejection) and type of nephritis (IgA nephropathy, focal segmental glomerular sclerosis and hemolytic-uremic syndrome) (all P < 0.05). In patients with de novo nephritis, the survival rate was significantly associated with cold ischemia time (≥12 h), immunosuppressive regime, recovery time of postoperative Scr (≥14 d) and complications at postoperative 1 month (acute renal tubular necrosis, ultra-acute rejection and acute rejection) (all P < 0.05). Conclusions The survival rate of patients with recurrent renal nephritis is lower than those in their counterparts with de novo nephritis and without nephritis. Cold ischemia time, immunosuppressive regime, recovery time of postoperative Scr and complications at postoperative 1 month are pivotal influencing factors of the clinical prognosis of patients with recurrent and de novo nephritis of the renal allograft.

Real-time assessments of liver quality in cold ischemia injury based on the technique of laser scanning confocal microscope / 中华器官移植杂志

Objective To assess the quality of donor liver allografts by employing laser scanning confocal microscope (LSCM ) and clinical liver function tests .Methods Sprague-Dawley rats were used for establishing cold ischemia models of liver allografts .According to different timepoints of cold ischemia ,four groups of CIT1h ,CIT6h ,CIT12h and CIT24h were designated .At the end of cold ischemia time (CIT) of each group , perfusion and preservation fluids were collected and fluoresceins perfused . After LSCM examinations ,tissue samples were harvested for HE examination .Finally a comparison was made between LSCM results and hematoxylin & eosin (HE) examinations .Also some relevant clinical parameters were detected in preserving and flushing fluids .Results Both LSCM examination and pathological examination indicated that the quality of liver allografts decreased significantly with the elapsing of time . Only the difference of LDH was statistically significant (P<0 .001) .Conclusions LSCM may be used for evaluating the ex vivo qualities of liver allografts .Simple handling and time efficiency re great advantages of LSCM .As compared with alanine transaminase (ALT ) and aspartate transaminase (AST ) ,LDH is a better indicator reflecting the quality of liver allografts .

Effect of hydrogen preconditioning during cold ischemia phase on activity of Nrf2 in rat pulmonary microvascular endothelial cells subjected to hypoxia-reoxygenation / 中华麻醉学杂志

Objective To evaluate the effect of hydrogen preconditioning during cold ischemia phase on the activity of nuclear factor erythroid 2-related factor 2 ( Nrf2) in rat pulmonary microvascular en-dothelial cells ( PMVECs) subjected to hypoxia-reoxygenation ( H/R) . Methods PMVECs were isolated from clean-grade male Sprague-Dawley rats, aged 2-3 weeks, using the tissue block adherence method and divided into 4 groups ( n=25 each) using a random number table method: control group ( group C) , H/R group, oxygen group ( O group) and hydrogen group ( H group) . Cells were incubated for 4 h with 4℃ low potassium dextransolution ( LPD) pre-equilibrated with 95％ oxygen and 5％ carbondioxide to simulate the cold ischemia phase. LPD pre-balanced with 95％ oxygen and 5％ carbon dioxide was replaced with LPD, and then cells were incubated for 1 h at room temperature to simulate the lung transplantation period. LPD was rapidly replaced with 37℃ M199 complete culture solution, and cells were incubated in the mixture of 40％ oxygen-5％ carbondioxide-55％ nitrogen to simulate the reperfusion period. In O and H groups, the cells were exposed to 40％ oxygen-60％ nitrogen and 3％ hydrogen-40％ oxygen-57％ nitrogen during the cold ischemia period, respectively, and the gas mixture was replaced every 20 min. The cell culture fluid was collected 4 h later for determination of interleukin ( IL )-6, IL-10 and tumor necrosis factor-alpha ( TNF-α) concentrations ( by enzyme-linked immunosorbent assay) and malondialdehyde ( MDA) concen-trations ( by thiobarbituric acid method) . The cytoplasm and nucleoproteins were extracted for measurement of Nrf2 expression ( by Western blot) and cell apoptosis ( by flow cytometry and TUNEL assay) . The cell apoptosis rate was calculated. Results Compared with C group, the IL-6, TNF-α and MDA levels were significantly increased, the IL-10 level was decreased, the apoptosis rate was increased, and the expres-sion of Nrf2 in nucleus was up-regulated in H/R group ( P＞0. 05) . Compared with H/R group, the IL-6, TNF-α and MDA levels were significantly decreased, the IL-10 level was increased, the apoptosis rate was decreased, and the Nrf2 expression in cytoplasm was down-regulated in O and H groups (P＜0. 05), the Nrf2 expression was significantly up-regulated in H group ( P＜0. 05) , and no significant change was found in the expression of Nrf2 in nucleus in O group ( P＞0. 05) . Compared with O group, the IL-6, TNF-αand MDA levels were significantly decreased, the IL-10 level was increased, the apoptosis rate was decreased, the expression of Nrf2 in nucleus was up-regulated, and the expression of Nrf2 protein in cytoplasm was down-regulated in H group ( P＜0. 05 ) . Conclusion The mechanism by which hydrogen preconditioning during cold ischemia phase reduces H/R injury to rat PMVECs is related to activating Nrf2 and thus inhibi-ting oxidative stress.

Correlation between the main indicators of organ donation and donor liver and the early prognosis of transplant after the death of citizens / 中华器官移植杂志

Objective To explore the correlation between main indicators of donor liver and early prognosis after liver transplantation.Methods The clinical data of 166 donors and recipients of post-mortem organ donation (DD) from June 2017 to June 2018 were retrospectively analyzed.The effects of donor age,sex,body mass index,serum sodium level,total bilirubin,prothrombin time and international standardized ratio on early allograft dysfunction (EAD) in liver transplant recipients were investigated.According to the culture results of donor liver preservation solution,the results were divided into positive group and negative group.Combined with the culture results of blood,sputum and drainage fluid after liver transplantation,the early infection rate of recipients in the two groups was observed.Results Univariate analysis showed that preoperative donor bilirubin total ＞17.1 mmol/L and donor cold ischemia time ＞8 h were risk factors for postoperative EAD in transplant recipients.Multivariate analysis showed that donor cold ischemia time ＞8 h was an independent risk factor for postoperative EAD in liver transplant recipients;the incidence of EAD in the group with cold ischemia time ＞8 h was significantly higher than that in the group with cold ischemia time ≤8 h (26.3％ vs.7.0％;P =0.003).The positive rate of postoperative sputum culture and drainage fluid culture in the donors with positive donor culture was 43.9％ and 48.8％,respectively,which was significantly higher than that in the negative group (10.7％ and 13.1％).The difference was statistically significant (P =0.000,P =0.000).The positive rate of postoperative blood culture in the positive group and the negative group was 12.2％ and 6.0％ with the difference being not statistically significant (P =0.161).Conclusion Cold ischemia time of the donor ＞8 h is an independent risk factor for EAD in recipients after liver transplantation.Shortening the cold ischemia time of donor liver can reduce the incidence of postoperative EAD in recipients.The culture results of preservation solution have a certain guiding effect on the postoperative anti-infective treatment of the recipients.

Effects of cold ischemia time on hepatic allograft function / Efeitos do tempo de isquemia fria sobre os enxertos hepáticos

ABSTRACT Background : Cold ischemia time is related to success of liver transplantation. Aim : To compare the impact of cold ischemia time on allografts locally collected to those collected distantly. Methods : Were evaluated 83 transplantations. The patients were divided in two groups: those who received liver grafts collected from cities out of Curitiba (n=42) and locally (n=41). From the donors were compared: cause of death, days at ICU, cardiac arrest, vasoactive drugs, lab exams, gender, age, and BMI. Were compared the subsequent information of receptors: cold ischemia time, warm ischemia time, length of surgery, lab exams, etiology of cirrhosis, MELD score, age, gender, histology of graft, use of vasoactive drugs, and blood components transfusion. Were evaluated the correlation between cold ischemia time and lab results. Results : The liver grafts collected from other cities were submitted to a longer cold ischemia time (500±145 min) compared to those locally collected (317,85±105 min). Donors from other cities showed a higher serum sodium level at donation (154±16 mEq/dl) compared to those from Curitiba (144±10 mEq/dl). The length of cold ischemia time was related to serum levels of ALT and total bilirubin. Conclusion : Liver grafts distantly collected underwent longer cold ischemia times, although it caused neither histologic injuries nor higher transfusion demands. There is a correlation between cold ischemia time and hepatic injury, translated by elevation of serum ALT and total bilirubin levels.

RESUMO Racional : O tempo de isquemia fria está relacionado ao sucesso do transplante hepático. Objetivo : Comparar o impacto do tempo dela sobre enxertos captados localmente com os distantes. Métodos : Avaliaram-se 83 transplantes. Os pacientes foram divididos em dois grupos: enxertos captados fora de Curitiba (n=42) e captados localmente (n=41). Dos doadores compararam-se causa do óbito, dias de UTI, parada cardíaca, drogas vasoativas, exames laboratoriais, gênero, idade e IMC. Dos receptores seguintes dados: tempos de isquemia fria e morna, tempo operatório, exames laboratoriais, causa da cirrose, MELD, idade na operação, gênero, biópsia do enxerto, uso de drogas vasoativas e necessidade de transfusões. Foi realizada avaliação de correlação entre o tempo de isquemia fria e os exames laboratoriais. Resultados : Os enxertos captados à distância foram submetidos a maior tempo de isquemia fria (500,3±145 min) quando comparados aos captados localmente (317,85±105 min). Os doadores de fora apresentaram níveis mais elevados de sódio no momento da doação (154±16 mEq/dl) comparados aos doadores de Curitiba (144±10 mEq/dl). Houve correlação entre o tempo de isquemia fria e os níveis de ALT e de bilirrubina total. Não houve diferenças ao comparar-se os demais dados. Conclusão : Enxertos captados à distância sofreram maior tempo de isquemia fria. Isso não refletiu nos prejuízos histológicos nem na demanda transfusional durante o pós-operatório. Houve correlação entre o tempo de isquemia fria e o grau de lesão hepática avaliada pela ALT e pela bilirrubina total.

Influence of sulforaphane preconditioning in myocardial cold ischemia-reperfusion injury through PI3K/Akt signaling pathway in rats / 吉林大学学报(医学版)

Objective:To explore the effect of sulforaphane (SFN) preconditioning on the cold myocardial ischemia-reperfusion injury (IRI) in the rats through PI3K/Akt signaling pathway.Methods:Sixty-four health male Sprague-Dawley (SD) rats were randomly divided into cold IRI group,SFN group,LY (LY294002) + cold IRI group,and LY+SFN group (n=16).The allogeneic heterotopic heart transplantation model was established by donor heart into recipient abdomen.The myocardium tissue was taken 24 h after reperfusion for the detection of histological changes using HE staining.The expression levels of Akt,p-Akt,Bax and Bcl-2 proteins were detected by immunohistochemistry and Western boltting methods.Results:The morphological results showed that the myocardium tissue damage was serious in cold IRI group and LY+cold IRI group,it was light in SFN group;the myocardium tissue damage of the rats in SFN+ LY group was ranged between cold IRI group and SFN group.Compared with IRI group,the expression levels of p-Akt protein and Bcl-2 protein in SFN group were increased (P＜0.05),and the expression level of Bax protein was decreased (P＜0.05).After treatment of blockage LY294002,compared with LY-+-cold IRI group,the expression level of p-Akt protein in LY-+-SFN group was not statistically significant (P＞0.05),the expression level of Bcl2 protein was increased (P＜0.05),the expression levels of Bax protein was decreased),and the ratio of Bcl-2/Bax was also increased (P＜0.05).Conclusion:SFN may attenuate cold IRI of heart transplantation through PI3K/Akt signaling pathway in the rats.

Influence of sulforaphane preconditioning in myocardial cold ischemia-reperfusion injury through PI3K/Akt signaling pathway in rats / 吉林大学学报(医学版)

Objective:To explore the effect of sulforaphane (SFN) preconditioning on the cold myocardial ischemia-reperfusion injury (IRI) in the rats through PI3K/Akt signaling pathway.Methods:Sixty-four health male Sprague-Dawley (SD) rats were randomly divided into cold IRI group,SFN group,LY (LY294002) + cold IRI group,and LY+SFN group (n=16).The allogeneic heterotopic heart transplantation model was established by donor heart into recipient abdomen.The myocardium tissue was taken 24 h after reperfusion for the detection of histological changes using HE staining.The expression levels of Akt,p-Akt,Bax and Bcl-2 proteins were detected by immunohistochemistry and Western boltting methods.Results:The morphological results showed that the myocardium tissue damage was serious in cold IRI group and LY+cold IRI group,it was light in SFN group;the myocardium tissue damage of the rats in SFN+ LY group was ranged between cold IRI group and SFN group.Compared with IRI group,the expression levels of p-Akt protein and Bcl-2 protein in SFN group were increased (P＜0.05),and the expression level of Bax protein was decreased (P＜0.05).After treatment of blockage LY294002,compared with LY-+-cold IRI group,the expression level of p-Akt protein in LY-+-SFN group was not statistically significant (P＞0.05),the expression level of Bcl2 protein was increased (P＜0.05),the expression levels of Bax protein was decreased),and the ratio of Bcl-2/Bax was also increased (P＜0.05).Conclusion:SFN may attenuate cold IRI of heart transplantation through PI3K/Akt signaling pathway in the rats.

Effects of edaravone on JAK2/STAT3 signaling pathway in transplanted donor liver of rats with different cold ischemia times / 天津医药

Objective To investigate the effect of edaravone on the JAK2/STAT3 signaling pathway after ischemia-reperfusion injury in donor rat liver under different cold ischemia times. Methods A total of 102 SD rats were randomly divided into sham operation group,control group and experimental group. Six rats were in sham operation group with free liver operation and no transplantation. Forty-eight rats were in control group and experimental group respectively, and divided into subgroups according to the different cold ischemia times (30 min, 6 h, 12 h and 18 h). There were 6 donors and 6 recipients in each group. The rat model of orthotopic liver transplantation was established by modifiedtwo cuff method. All the donors were perfused by abdominal aorta and the warm ischemia time was 3-5 min. After different cold ischemia times, the experimental group was treated with edaravone (3 mg/kg) at 5 min before the opening of the new hepatic artery, and control group was injected with 3 mg/kg saline. Recipients of each group were sacrificed after 6 h. Finally, real-time fluorescence quantitative PCR was used to analyze the relative expression of JAK2/STAT3 mRNA of donor liver. Results The GAPDH gene and JAK2/STAT3 were well amplified. Under the same cold ischemia time, compared with the control group, the relative expression of JAK2/STAT3 was significantly decreased in the experimental group (P<0.05). With the prolongation of cold ischemia time, the relative expressions of JAK2 and STAT3 mRNA showed a decreasing trend in control group and experimental group, while the relative expression of JAK2 mRNA increased first and then decreased in the experimental group (P<0.05). Conclusion Edaravone has a protective effect on transplanted donor liver during different cold ischemia times, and extends the cold ischemia time for 18 h, which may be related to the inhibition of JAK2/STAT3 signal transduction pathway.

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

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

Effect of hypothermia status in donors on renal graft function after renal transplantation from donation after citizen's death / 器官移植

Objective To evaluate the effect of hypothermia status in the donors upon the renal graft function after renal transplantation from donation after citizen's death. Methods Thirty-six eligible donors were randomly divided into the normal temperature (body temperature 36.5-37.5 ℃ , n=19) and hypothermia groups (body temperature 34.0-35.0 ℃ , n=17). The matched recipients undergoing renal transplantation were also assigned into the normal temperature (n=38) and hypothermia groups (n=34). Perioperative conditions of the donors and recipients were compared between two groups. And postoperative renal graft function of the recipients were statistically compared between two groups, including the incidence of delayed graft function (DGF) and primary nonfunction (PNF). Results No statistical significance was identified in the perioperative amount of urine volume, serum creatinine (Scr), systolic blood pressure, saturation oxygen, warm ischemia time and cold ischemia time of the donors between two groups (all P>0.05). No statistical significance was noted in terms of the operation time, intraoperative mean blood glucose and intraoperative mean arterial pressure of the recipients between two groups (all P>0.05). Postoperative incidence of DGF of the recipients in the hypothermia group was 6%, significantly lower than that in the normal temperature group (24%) (χ2=4.393, P=0.036). Postoperative incidence of PNF of the recipients was 3% in both the hypothermia and normal temperature groups with no statistical significance (χ2=0.000, P=1). Conclusions The hypothermia status of the donors can significantly reduce the incidence of DGF, whereas exerts no evident effect upon the incidence of PNF in the recipients.