Chinese expert consensus on organ protection of transplantation (2022 edition) / 器官移植

Organ transplantation is the most effective treatment for end-stage organ failure, and voluntary donation after citizen's death is the only source of transplant organ in China. Clinically, transplant organ protection technique plays a critical role in improving the quality of transplant organs and the prognosis of recipients. On the basis of domestic and worldwide basic research and clinical practice of transplant organ protection and according to the Oxford evidence classification and GRADE system, the experts organized by Branch of Organ Transplant Physicians of Chinese Medical Doctor Association, Branch of Transplantation Group of Surgery of Chinese Medical Association and China Liver Transplant Registry Scientific Committee had compiled and published the Chinese Expert Consensus on Organ Protection of Transplantation (2016 edition) for liver, kidney, pancreas, small intestine, heart, lung transplant organs. With the support of China Liver Transplant Registry, National Trauma Medical Center, National Quality Control Center for Human Donated Organ Procurement, National Clinical Research Center for Orthopedics, Sports Medicine & Rehabilitation and National Center for Healthcare Quality Management in Liver Transplant combined with recent domeatic and worldwide clinical practice and research progress for organ transplantation and organ protection, the Chinese Expert Consensus on Organ Protection of Transplantation (2022 edition) has been published recently. This consensus focuses on updating the technical progress and evidence-based medicine of organ procurement, preservation, transport, and quality evaluation in clinical practice. Additionally, the content of composite tissue transplantation mainly including limb transplantation has also been covered. The aim is to promote the the scientific and standardized clinical organ transplantation.

Human cornea conservation in coconut water solution / Conservação de córneas humanas em solução de água de coco

ABSTRACT Purpose: The aim of this study was to evaluate the physical and chemical characteristics of coconut water and to analyze the use of coconut water solution for the conservation of human corneas. Methods: This was an experimental and controlled study performed at the Eye Bank of the General Hospital of Fortaleza. The coconut water-based solution was prepared at the Goat Seed Technology Laboratory of the Department of Veterinary Medicine of the State University of Ceará. Discarded corneas from the Eye Bank were divided into two groups for sequential experiments: G1, coconut water-based solution (experimental group), and G2, conservative treatment with OPTISOL GS® (control group). The osmolality of corneas in G1 was analyzed sequentially at 275, 300, 325, 345, 365, and 400 mOsm/L. The viability of the corneas was determined by specular microscopy and biomicroscopy on the first, third, and seventh days. Results: Corneas preserved in a solution of 365 and 345 mOsm/L had a transparency of 8 mm until the third day and had diffuse edema in the periphery, central folds, and partial epithelium loss until the seventh day. The 365-mOsm/L solution was associated with the worst results during follow-up. Corneas placed in Optisol-GS retained their original aspects. Conclusions: Coconut water-based preservative partially maintained corneal transparency and epithelial integrity, especially during the first three days of follow-up. The coconut water-based solutions used were not effective for use as preservatives in a human eye bank.(AU)

RESUMO Objetivos: As características físico-químicas e o baixo custo da água de coco foram fundamentais para o este estudo. Analisar o uso de solução a base de água de coco como meio de conservação de córneas humanas em banco de olhos. Métodos: Estudo experimental e controlado realizado no Banco de Olhos do Hospital Geral de Fortaleza. Utilizou-se solução à base de água de coco preparada no laboratório de Tecnologia de Sêmen de Caprinos do Departamento de Medicina Veterinária da Universidade Estadual do Ceará. Foram usadas córneas de descartes divididas em dois grupos: G1 (Conservante com água de coco) - grupo experimental e G2 (grupo Conservante com OPTISOL GS®) grupo controle, em experimentos sequenciais. A osmolaridade do G1 foi analisada sequencialmente com 275, 300, 325, 345, 365 e 400 mOsm/L. A viabilidade das córneas foram realizadas por microscopia especular e biomicroscopia nos 1º, 3º e 7º dias. Resultados: As córneas em solução de 365 e 345 mOsm/L apresentavam transparência nos 8mm centrais até o 3º dia, com edema em toda periferia, dobras centrais e edema 2+, com perda parcial do epitélio até 7º dia, sendo o de maior osmolaridade com melhor transparência durante o seguimento. Grupo com 275, 300 e 400 mOsm/L, córnea opaca, edema difuso, perda total do epitélio no 3º dia. As córneas em Optisol mantiveram seus aspectos. Conclusões: O conservante à base de água de coco manteve em parte a transparência corneana e a integridade epitelial, especialmente nos primeiros 3 dias de seguimento. A solução conservante com água de coco nas formulações utilizadas não se mostrou eficaz para o uso em banco de olhos humanos.(AU)

Study of isolation efficiency of new islet isolation solution for mouse islets / 器官移植

Objective To investigate the isolation and protective effect of a new islet purification solution (IPS)-Optiprep solution on the islet in mouse models. Methods The digested pancreatic islets were divided into the IPS and UW groups according to the islet volume. The pancreatic islets were isolated by the continuous gradient density centrifugation using IPS-Optiprep or UW-Optiprep solutions. The purification efficiency and isolated islet activity of purification solution were compared between two groups. The diabetic mouse models were successfully induced and randomly assigned into three groups. In the experimental group (n=10), the mice received pancreatic islet transplantation using islets isolated and purified by the IPS-Optiprep solution. In the control group (n=10), the mice underwent pancreatic islet transplantation using islets isolated and purified by the UW-Optiprep solution. In the sham surgery group (n=5), the mice merely underwent surgery without pancreatic islet transplantation. Postoperative blood glucose levels were detected and compared among three groups. The blood glucose levels of intraperitoneal glucose tolerance test at postoperative 21 d were statistically compared between the experimental and control groups. The cost of the preparation of two isolation solutions was also compared. Results Compared with the UW group, the islet equivalent (IEQ), islet purity, recovery rate and islet integrity were significantly higher in the IPS group. Islet morphological observation revealed that the islet membrane was complete and the islet diameter in the IPS group was considerably larger than that in the UW group. The activity of purified islets in the UW group was significantly higher than that in the IPS group [(88±5)% vs. (84±3)%, P<0.01]. Compared with the UW-Optiprep solution, identical in vivo islet function was obtained in the IPS-Optiprep solution.The cost of IPS-Optiprep solution was significantly less than that of the UW-Optiprep solution. Conclusions The new IPS-Optiprep solution yields higher islet isolation efficiency, purification, integrity and recovery rate and significantly reduces the purification cost compared with the UW-Optiprep solution. Nevertheless, IPS-Optiprep solution exerts a less protective effect on the activity of islet cells, which is probably correlated with the high islet integrity and the endotoxin in the IPS-Optiprep solution.

Contrastive study on cryopreservation of rat amputated limb with UW solution and HTK solution / 中华显微外科杂志

Objective To compare the effect on preserving rat amputated extremities during the cold preservation between the UW solution and HTK solution.Methods Thirty healthy adult Wistar rats were randomly divided into 3 groups (10 rats in each group) for producing the models of amputated limbs of rats.The three groups were separately irrigated the amputated limbs with UW solution,HTK solution and saline from femoral artery.After irrigation,the extremities were preserved in the refrigerator at 4 ℃.Samples of skeleton muscle tissue were taken for pathological and biochemical examination every 6 hours since the amputation.Results After perfusion,HE:changes of organizational structures of skeletal muscle were lighter in experimental groups than in control group (saline-perfusion group) at the same time.While samples in UW solution-perfusion group has lighter changes of organizational structure than those in HTK solution-perfusion group.TEM:at 12 h,all the three groups have shown the mitochondria edema.At 24 h,the myofibril in saline-perfusion group become fragments.Biochemical examination:at 24 h,indexes in salineperfusion group,HTK solution group and UW solution group were:ALP (U/L) 3.62 ± 1.32,3.37 ± 0.84 and 2.68 ± 1.59,respectively;ALT(U/L) 542.25 ± 129.36,521.82 ± 97.32 and 462.53 ± 74.18,respectively;AST (U/L) 2200.12 ± 687.61,2002.20 ± 632.27 and 1742.87 ± 396.09,respectively;CK(U/L) 313190.83 ± 42041.32,283173.50 ± 31488.87 and 271319.67 ± 41147.52,respectively;LDH (U/L) 32409.50 ± 4253.20,30382.50 ± 2337.18 and 30047.83 ± 4628.78,respectively;GLU(mmol/L) 0.242 ± 0.041、0.240 ± 0.044 and 0.252 ± 0.049,respectively;LAC (mmol/L) 1.790 ± 0.160,1.792 ± 0.196 and 1.993 ± 0.366,respectively;SOD(U/mL) 80.82 ± 9.46,91.62 ± 14.97 and 73.71 ± 10.60,respcctively.There were statistical difference among the UW solution-perfusion group and centrol group in ALP and GLU (P ＜ 0.05).UW solution-perfusion group has better results than HTK solution-perfusion group in SOD (P ＜ 0.05).Conclusion Amputated limbs irrigated with organ preservation solution as a way to protect skeletal muscle had great effects on lightening tissue damage and keeping tissue active.Between the two usual organ preservation solution,UW solution had a stronger protective effects on the amputated limbs than HTK solution.

Effect of heart preservation solution containing pinacidil on mitochondrial function in isolated rat hearts / 中华麻醉学杂志

Objective To investigate the effect of heart preservation solution containing pinacidil on mitochondrial function in isolated rat hearts. Methods One hundred and twenty pathogen-free SD rats of both sexes weighing 250-350 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 65 mg/kg. Their hearts were immediately removed and perfused in a Langendorff apparatus. Left ventricular enddiastolic pressure was measured from a fluid-filled latex balloon inserted in the left ventricle. The isolated hearts were randomized into 5 groups (n = 24 each):group Ⅰ was perfused with cardioplegic solution HTK; group Ⅱ with HTK containing pinacidil (a non-specific sarcKATP and mitoKATP channel opener) 0.5 mmol/L; group Ⅲ with HTK containing pinacidil 0.5 mmol/L + 5-HD (a selective mitoKATP channel blocker) 100 μmol/L; group Ⅳ with HTK containing pinacidil 0.5 mmol/L + HMR-1098 100 μmol/L (a selective sarcKATP channel blocker) and group Ⅴ with HTK containing pinacidil 0.5 mmol/L + 5-HD 100 μmol/L + HMR-1098 100μmol/L. The isolated hearts were perfused with simple HTK or HTK containing pinacidil or pinacidil + 5-HD and/or HMR 20 ml/kg at 10 ml/min and then removed from Langendorff apparatus and dipped into the same HTK solution for 8 h at 4 ℃followed by 60 min reperfusion. The respiratory function of mitochondria (respiratory control rate (RCR), the rate of oxygen consumption in state 3/state 4 and P/O) was measured at the end of equilibration (T1) after 8 hpreservation (T2) and at the end of 60 min reperfusion (T3). The CK-MB and LDH activities and cTnI expression in myocardium was detected at T1 and T3. The ultrastructure of myocardium was examined at T3. Results Perfusion suspension-reperfusion (PS/R) significantly decreased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and increased myocardial cTnI concentration and CK-MB and LDH activities at T3 compared with baseline at T1 in group Ⅰ. Pinacidil significantly increased mitochondrial respiratory function (RCR, P/O and rate of O2 consumption in state 3) and decreased myocardial cTnI concentration and CK-MB and LDH activities in group Ⅱ as compared with group Ⅰ-indicative of protective effect of pinacidil on mitochondria against PS/R injury. The protective effect of pinacidil against PS/R injury was attenuated by 5-HD and/or HMR1098. The myocardial damage was slightest in group Ⅱ . Conclusion Both sarcolemmal and mitochondrial KATPchannel are involved in the protective effect of pinacidil against PS/R-induced myocardial damage during heart preservation.

Observation on cold-stored rat Hver flushed with self-designed multi-organ preservation solution / 中华消化外科杂志

Objective To study the effect of serf-designed multi-organ preservation solution(SMO)on cold-stored rat liver.Methods The rat livers were preserved with SMO solution(group A,n=15),UW solution(group B,n=15)and HC-A solution(group C,n=15),respectively.The livers were transplanted orthotopically after 6-,12-,24-hour preservation.The changes of liver function at hour 12 after transplantation were detected and conditions of the survived rats at day 14 after transplantation were observed.Results There was no morphological change of the livers in group A within 24 houm.The total bilimbin,alaninetransaminase,and hyaluronic acid increased in group A and B as the preservation time increased,but the levels of the 3 indexes of group A were significant lower than those in group C(F=49.027,70.280,34.349,71.532,446.544,303.408,P＜0.05).No rat djed in group A 14 days after liver transplantation.Conclusions SMO and UW solution have similar effect on protecting the liver.and the protective effect of the 2 solutioas was better than that of HC-A solution.

Experimental Study on the Cryopreservation of LLC-PK1 Epithelial Cells with Hypoxic UW Solution / 华中科技大学学报（医学）（英德文版）

The effects of oxygen partial pressure on cryopreservation of the cells with organ preservation solution were explored. Hypoxic UW solution was made by purging the UW solution with argon. The pig proximal tubule epithelial cells (LLC-PK1 cells) were cryopreserved in hypoxic UW solution (Ar-UW group) or standard UW solution (UW group) at 4℃ for 48 h. Trypan blue staining and LDH detection were performed to evaluate the injury of the cells. The results showed that the oxygen partial pressure in Ar-UW group was significantly declined from 242±6 mmHg to 83±10 mmHg. After cryopreservation at 4℃ for 48 h, LDH leakage rate and Trypan blue-stained rate in Ar-UW group were (11.3±3.4)% and (10.5±4.7)%, respectively, which were significantly lower than in UW group [(49.5±6.9)% and (47.6±9.3)% respectively, both P＜0.01]. It was concluded that lower oxygen partial pressure of UW solution was more beneficial to the cryopreservation of LLC.

Preparation and Preservation of Hypoxia UW Solution / 华中科技大学学报（医学）（英德文版）

In order to explore the method to prepare hypoxia UW solution and the stability and preservation of hypoxia UW solution, UW solution was purged by argon or air for 15 min or 60 at a flow rate of 0.8 or 2 L/min, and the oxygen partial pressure of UW solution was detected. The hy-poxia UW solution was exposed to the air or sealed up to preserve by using different methods, and the changes of oxygen partial pressure was tested. The results showed that oxygen partial pressure of 50 mL UW solution, purged by argon for 15 min at a flow rate of 2 L/min, was declined from 242±6 mmHg to 83±10 mmHg. After exposure to the air, oxygen partial pressure of hypoxia UW solution was gradually increased to 160±7 mmHg at 48 h. After sealed up by the centrifuge tube and plastic bad filled with argon, oxygen partial pressure of hypoxia UW solution was stable, about 88±13 mmHg at 72 h. It was concluded that oxygen of UW solution could be purged by argon efficiently. Sealed up by the centrifuge tube and plastic bag filled with argon, oxygen partial pressure of UW so- lution could be stabilized.