| Background Since the start of organ donation work,the number of kidney transplantation cases in China has been increasing year by year.With the rapid development of kidney transplantation work,there has been a huge imbalance between the number of donated kidneys and renal failure patients.In order to shorten the waiting time of patients with renal failure as much as possible,the Donor kidney standard had to be expanded to increase the Donor kidney source.However,while improving the quality of life of more patients with chronic renal failure,it also brought a new problem,namely donor-derived Infection(DDI).DDI refers to the organ donation,the donor of pathogens in the body through the transplant process make the recipients suffering from the same infection,DDI incidence of complications and mortality increased significantly,the involvement of the recipients of renal transplantation and other solid organ transplant of DDI,especially drug-resistant bacteria infection risk spurt,grim,explore the effective methods to donor source of infection prevention and control,is imminent,the resources to make full use of the hard-won organs and improve the prognosis of renal transplantation has important significance.Part I The bactericidal effectiveness of cefoperazone and sulbactam under the condition of in vitro low temperature perfusionObjective In order to block the transmission of pathogens causing donor-derived infection and reduce the incidence of infection after kidney transplantation.Experiments were designed to investigate whether cefoperazone and sulbactam was still effective against sensitive pathogens in vitro when mixed with organ preservation solution at low temperature.Methods Three kinds of bacteria commonly found in organ lavage fluid: Klebsiella pneumoniae,Acinetobacter baumannii and Pseudomonas aeruginosa were selected to make bacterial suspensions respectively.Each bacterium was inoculated and incubated for 20 hours in brain-heart Infusion medium(BHI)at 5 ° C and 37 ° C respectively.Each bacterium was then inoculated and cultured in KPS-1 organ preservation solution at 5℃ and 37℃ for 20 hours,respectively.The growth curves of each bacterium under different conditions were plotted.Last each bacteria inoculation and cultivation in 5 ℃ containing different concentrations of cefoperazone sodium sodium shu ba jotham KPS-1 20 hours in organ preservation solution,understanding of cefoperazone sodium sodium shu ba jotham the minimal Inhibitory concentrations(MIC)of each of the bacteria in vitro(Minimum Inhibitory Concentration,MIC),draw the antibiotics-to the time of each type of bacteria sterilization curve,determine the time necessary to kill three kinds of bacteria.Results At 5℃,the three kinds of bacteria were cultured in KPS-1 organ preservation solution and in KPS-1 organ preservation solution for 20 hours,respectively.The growth of the bacteria was relatively slow in the former while no obvious growth was observed in the latter.At 37℃,the three kinds of bacteria were cultured in BHI and KPS-1 organ preservation solution for 20 hours,respectively.In the former,the increase of bacteria was rapid,while in the latter,the change of the number of bacteria was not obvious or even decreased.MIC values of cefoperazone sulbactam sodium against three bacteria inoculated in organ preservation solution at 5℃ were 92ug/ m L for Klebsiella pneumoniae,103ug/ m L for Acinetobacter baumannii,and 85ug/ m L for Pseudomonas aeruginosa.The three bacteria were inoculated and cultured in the KPS-1 organ preservation solution containing cefoperazone sodium and sulbactam sodium with a concentration of 0.5 times MIC to 16 times MIC.It was found that when the drug concentration was greater than 8 times MIC and the duration was greater than 8 hours,the killing efficiency of the three bacteria could reach more than 99%.ConglusionsThe growth curves of the three bacteria under various conditions were compared,and it was found that KPS-1 organ preservation solution itself had almost no effect on the growth of the three bacteria at 5℃ or 37℃,and the preservation solution itself would not promote the growth of bacteria or kill the three bacteria.When the concentration of cefoperazone sodium and sulbactam sodium was more than 1000ug/ml at 5℃ and KPS-1 organ preservation solution was added for more than 8 hours,the killing effect of three kinds of bacteria was observed.Part 2 The bactericidal safety of cefoperazone and sulbactam under in vitro low temperature perfusionObjective An animal model was established to preliminarily evaluate the safety of cefoperazone sodium and sulbactam sodium directly acting on the donor kidney through mechanical perfusion,and to explore effective and safe preventive measures against donor-derived infection,with a view to further popularizing its use in clinical practice.Methods Thirty adult Beagle dogs were randomly divided into drug addition group and drug free group,with 15 dogs in each group.The left kidney was removed in both groups and the left kidney was removed after the same period of low temperature mechanical perfusion,and then the left kidney was autograft and the right kidney was removed.Cefoperazone sodium and sulbactam sodium were added during mechanical perfusion in dosing group.No antibiotics were added during mechanical perfusion in drug free group.The serum creatinine and urea nitrogen of the two groups of experimental dogs were measured by taking venous blood samples at 8 am on the 1st,2nd,3rd,5th,7th,10 th,13th,16 th,19th,22 nd,25th and 28 th days after surgery.The serum creatinine and urea nitrogen were compared by two independent samples t-test between the treated and non-treated Beagle dogs within 28 days after surgery.P < 0.05 was considered statistically significant.After 28 days,all the experimental dogs were sacrificed and dissected to compare the pathological changes of the left kidney between the two groups.Results The two experimental dogs in the dosing group were euthanized on the 5th and 7th day after surgery,and ultrasound examination revealed renal artery thrombosis due to the continuous increase of creatinine without any improvement sign.Serum creatinine and urea nitrogen of the other 13 dogs in the drug addition group increased briefly on the second day after surgery,then gradually decreased and remained stable,and the levels of serum creatinine and urea nitrogen were all higher than normal levels on the first,second,third,fifth and seventh days after surgery.Two experimental dogs in the drug-free group showed continuous increase of creatinine without signs of improvement.Ultrasound found abnormal renal enlargement,and they were euthanized on the 9th day after surgery.The renal volume was found to be abnormally increased,showing edema and congestion,and the texture was soft.The serum creatinine and urea nitrogen levels of the other 13 dogs in drug-free group were higher than those before operation at 1,2,3,5 and 7 days after operation.There was no significant difference in serum creatinine and urea nitrogen levels between the two groups within 28 days after operation.(creatinine: t=-4.544,-5.083,-5.171,-4.946,-5.460,-5.345,-3.757,-3.918,-3.435,-1.996,-2.030,and-1.998,P = > 0.05;Urea nitrogen: t=-4.810,-8.119,-10.379,-12.789,-15.210,-9.983,-5.014,-4.599,-2.657,-0.603,0.711,0.689,P = > 0.05).After 28 days of observation,the dogs were euthanized.Anatomical analysis of the two groups showed that the size of the transplanted kidney was about 7 cm×4 cm×2 cm,the texture was moderate,there was intestinal encapsulation around the kidney,no thrombosis occurred in the renal artery and internal iliac artery,and the renal vein and external iliac vein returned normally.The blood flow of the reconstructed vessels was good,and no stenosis was observed at the ureter and bladder anastomotic site.Under light microscope,HE staining of transplanted kidney tissue of two groups of experimental dogs showed slight changes in renal tissue structure,uniform distribution of glomeruli,no abnormal changes in Baumann’s bursa.The epithelial cells of renal tubules were square or columnar,with abundant cytoplasm and intact lumen structure without obvious injury.Conglusions Therefore,it is feasible to use cefoperazone sodium and sulbactam sodium to prevent common donor-derived pathogens during low temperature mechanical perfusion,which lays a foundation for clinical application in the future. |
PDF Full Text Request