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OBJECTIVE@#To investigate the efficacy of a novel artificial perfusate based on oxygen-carrying perfluoronaphthalene-albumin nanoparticles in normothermic machine perfusion (NMP) for preservation of porcine liver donation after cardiac death.@*METHODS@#Artificial perfusate with perfluoronaphthalene-albumin nanoparticles was prepared at 5% albumin (w/v) and its oxygen carrying capacity was calculated. The livers of 16 Landrace pigs were isolated after 1 h of warm ischemia, and then they were divided into 4 groups and preserved continuously for 24 h with different preservation methods: cold preservation with UW solution (SCS group), NMP preservation by whole blood (blood NMP group), NMP preservation by artificial perfusate without nanoparticles (non-nanoparticles NMP group) and NMP preservation by artificial perfusate containing nanoparticles (nanoparticles NMP group). Hemodynamics, tissue metabolism, biochemical indices of perfusate and bile were monitored every 4 h after the beginning of NMP. Liver tissue samples were collected for histological examination (HE and TUNEL staining) before preservation, 12 h and 24 h after preservation.@*RESULTS@#The oxygen carrying capacity of nanoparticles in 100 mL artificial perfusate was 6.94 μL/mmHg (1 mmHg=0.133 kPa). The hepatic artery and portal vein resistance of nanoparticles NMP group and blood NMP group remained stable during perfusion, and the vascular resistance of nanoparticles NMP group was lower than that of blood NMP group. The concentration of lactic acid in the perfusate decreased to the normal range within 8 h in both nanoparticles NMP group and blood NMP group. There were no significant differences in accumulated bile production, alanine aminotransferase and aspartate aminotransferase in perfusate between nanoparticles NMP group and blood NMP group (all P>0.05). After 24 h perfusion, the histological Suzuki score in blood NMP group and nanoparticles NMP group was lower than that in SCS group and non-nanoparticles NMP group (all P<0.05), and the quantities of TUNEL staining positive cells in blood NMP group and non-nanoparticles NMP group was higher than those in nanoparticles NMP group and SCS group 12 h and 24 h after preservation (all P<0.05).@*CONCLUSION@#Artificial perfusate based on oxygen-carrying nanoparticles can meet the oxygen supply requirements of porcine livers donation after cardiac death during NMP preservation, and it may has superiorities in improving tissue microcirculation and alleviating ischemia-reperfusion injury.
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Animals , Swine , Liver Transplantation , Organ Preservation , Liver , Perfusion , Death , Oxygen/metabolismABSTRACT
Background & objectives: Standard donor lung preservation with cold flush and storage allows up to six hours between retrieval of lungs from the donor and transplantation in the recipient. Ex vivo lung perfusion (EVLP) systems mimic physiological ventilation and perfusion in the donor lungs with potential for prolonged lung preservation and donor lung reconditioning. In this study, it was aimed to perform EVLP on discarded donor lungs using a locally developed EVLP system. Methods: Equipment that are routinely used for cardiac surgeries were collected and a functional EVLP system was assembled. This system was used on five pairs of lungs retrieved from brain-dead organ donors. The lungs were ventilated and pulmonary circulation was continuously perfused with a solution containing oxygen and nutrients for four hours. The system was tested without red blood cells (RBCs) added to the solution (acellular group; n=3; A1, A2 and A3) and also with RBCs added to the solution (cellular group; n=2; C1 and C2). Results: The EVLP system was successfully used in four (A1, A2, A3 and C2) of the five lung pairs. Mechanical and gas exchange functions of the lungs were preserved in these lung pairs. One lung pair (C1) worsened and developed pulmonary oedema. Histopathological examination of all five lung pairs was satisfactory at the end of the procedure. Major challenges faced were leakage of solution from the system and obstruction to drainage of RBCs containing solution from the lungs. Interpretation & conclusions: The results of the present study suggest that, it is possible to maintain the lungs retrieved for transplantation in a physiological condition using a locally prepared EVLP system and a solution without RBCs.
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Invasive aspergillosis remains a problem in solid organs and haematopoietic stem cell transplants. We report a case of 12-year-old female with primary hyperoxaluria with regular haemodialysis for the end-stage renal disease. She underwent a combined liver and renal transplantation which got infected by aspergillosis. In this case study, it is speculated that the most likely source of Aspergillus was contaminated preservative solution (perfusate), resulting in infection within the donor kidney and subsequent systemic infection in the recipient. This case study calls for critical analysis and needs for the routine culture of the preservative solution before transplantation, to detect any fungal contamination and manage it prophylactically.
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OBJECTIVE:To establish a method for the simultaneous determination of the contents of Shuanghuanglian freeze-dried powder in placental perfusate. METHODS:HPLC was performed on the column of Agilent Zorbax-C18 with mobile phase of acetonitrile-1%formic acid aqueous solution(gradient elution)at flow rate of 1.0 ml/min,the internal standard was puera-rin,detection wavelength was 280 nm,column temperature was 25 ℃ and sample volume was 10 μl. RESULTS:There was a lin-ear range between linear ranges and peak area of 8 ingredients(r≥0.999 0);RSDs of within-day and inter-day precision tests were no more than 1.9%,repeatability tests was no more than 7.3%;average recoveries were in the range of 92.73%-112.37%(RSD=3.2%-8.2%,n=6);and average matrix effects were 90.33%-105.78%(RSD=3.2%-8.0%,n=6). CONCLUSIONS:The method is rapid,sensitive and specific,and can be used to the simultaneous determination of the contents of 8 ingredients of Shuan-ghuanglian freeze-dried powder in placental perfusate.
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Objective To observe the influence of nasal washing with Bidouguan Perfusate (BP) on nasal mucosal cytokines of nasal polyp patients after operation. Methods Thirty-nine nasal polyp patients were randomized into BP group ( N=18) and normal saline group ( N=21). All patients received operation for nasal polyp and postoperative comprehensive treatment, and additionally, BP group was given washing with BP and normal saline group was washed with 0.9%normal saline for 3 months. The scores of SNOT-20 and Lund-Kennedy were observed for the evaluation of therapeutic effect. Enzyme-linked immunosorbent assay ( ELISA) was used for detection of interleukin 5 (IL-5), interleukin 8 (IL-8), interleukin 10 (IL-10), interferon gamma (IFN-γ), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF-α) in nasal mucosa before operation and 3 months after operation. Results ( 1) SNOT-20 and Lund-Kennedy scores were decreased after operation ( P 0.05). ( 2) The expression levels of IL-5, IL-8, IL-10, IFN-γ, TNF-αand GM-CSF in the nasal mucosa were decreased in both groups 3 months after operation (P0.05). Conclusion Nasal washing with BP has similar effect to normal saline on improving the quality of life and nasal mucosal histology 3 months after operation, but BP has better effect on regulating nasal mucosal IL-5, IL-8, IL-10 and IFN-γ.
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OBJECTIVE: To develop a method for determining the concentration of lamivudine in placental perfusate by employing ultra fast liquid chromatography with tandem mass spectrometry (UFLC-MS/MS). METHODS: Famotidine was used as the internal standard. After protein precipitation with acetonitrile, the chromatography was performed on a Shim-pack XR-ODS II C18 column (2.0 mm × 75 mm, 2.2 μm) with mobile phase consisting of methanol and 10 mmol · L-1 ammonium acetate, gradient elution was used, and the flow rate was 0.4 mL · min-1. Tandem mass spectrometric detection was conducted by using multiple reaction monitoring (MRM) in positive ionization mode with an electrospray ionization interface. RESULTS: The calibration curve of lamivudine was linear in the range of 1-2500 ng · mL-1. The intra- and inter-batch RSDs were less than 10%. The method recovery was more than 72%. CONCLUSION: The method is rapid and accurate, which is suitable for the determination of lamivudine in placental perfusate.
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Objective:To study the effect of sinuses perfusate plus budesonide in chronic rhinitis and sinusitis after endoscopic surgery. Methods:Rhinitis and sinusitis patients were randomly divided into ffect of sinuses perfusate plus budesonide in chronic rhinitis and sinusitis after endoscopic surgery given sinuses perfusate and observation group given sinuses perfusate plus budesonide. Then saccharin clearance time, pulmonary function and inflammatory factor level were observed. Results:saccharin clearance time, heart rate, respiratory rate after walking, MRC score and IL1, IL2, IL4, IL6, IL8 levels of observation group were lower than control group;a second rate, 6 MWT were significantly higher than control group. Conclusion:Sinuses perfusate plus budesonide has positive clinical value for it can improve treatment effect and alleviate inflammatory state.
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OBJECTIVE: To establish a UPLC-MS/MS method for the determination of antipyrine in placental perfusate. METHODS: Sample was precipitated with acetonitrile. Analysis of antipyrine and estazolam(internal standard) were carried out on a Shim-pack XR-ODS II C18 column(2.0 mm × 75 mm, 2.2 μm). Gradient elution was adopted using acetonitrile and water (containing 0.1% formic acid, V/V) at a flow rate of 0.4 mL · min-1. Detection was performed with multiple reactions monitoring (MRM) using positive electrospray ionization(ESI) at m/z 189.2→56.2 for antipyrine and m/z 295.2→205.2 for estazolam. RESULTS: The calibration curve of antipyrine was linear over the range of 1.2 -1200 ng · mL-1 (r =0.9993). Inter- and intra-day RSDs were less than 15%, and the analytes were proven to be stable. CONCLUSION: This method is rapid, sensitive and specific, and can be applied to the determination of antipyrine in placental perfusate.
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PURPOSE: Reperfusion after acute ischemia is associated with high rates of morbidity and mortality due to the ischemic injury itself and the following myonephropathic metabolic syndrome, in spite of complete revascularization. Therefore, we attempted to verify the difference in reperfusion injury with or without pre-reperfusion intravascular rinsing in a canine acute limb ischemia model. METHOD: We used five female mongrel dogs. Complete acute hind limb ischemia was induced by infrarenal aortic and bilateral femoral artery clamping for 8 hours. We divided the dogs into two groups. In the rinse group (RG): three dogs were declamped with intravascular rinsing, and in the non-rinse group (non-RG): two dogs were declamped without any additional procedures. The perfusate solution was infused through both femoral arteries and was drained via both femoral veins. Serial blood samples were obtained four times. The gastrocnemius muscles were biopsied 3 days after reperfusion. RESULT: The results at preischemia, just before, 30 min, and 3 days after reperfusion were as follows: Creatine phosphokinase (CK) was 204, 1031, 1373, and 443 (IU/L) in the RG and 159, 3855, 6345, and 5455 (IU/L) in the non-RG. Lactate dehydrogenase (LDH) was 177, 248, 173, and 232 (IU/L) in the RG and 95, 508, 638, and 878 (IU/L) in the non-RG. Aspartate transaminase (AST) was 23, 50, 43, and 42 (IU/L) in the RG and 19, 118, 154, and 399 (IU/L) in the non-RG. Potassium (K) was 5.7, 5.8, 5.4, and 4.0 (mEq/L) in the RG and 5.4, 5.5, 5.7, and 5.3 (mEq/L) in the non-RG. Muscle injury in the non-RG was more severe than in the RG. CONCLUSION: There was a considerable difference between the groups in the CK, LDH and AST levels and the muscle biopsy findings. The results showed that the intravascular rinsing provided beneficial effects in reperfusion injury acute limb ischemia.
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Animals , Dogs , Female , Humans , Aspartate Aminotransferases , Biopsy , Constriction , Creatine Kinase , Extremities , Femoral Artery , Femoral Vein , Ischemia , L-Lactate Dehydrogenase , Mortality , Muscles , Potassium , Reperfusion , Reperfusion InjuryABSTRACT
Objective To establish a method for the determination of chlorogenic acid in Fuyanqing perfusate (FP). Methods The sample was extracted with methyl and the chromatographic condition were as follows: C18 chromatographic column (250 mm? 4.6 mm,5 ? m),a mobile phase of methyl alcohol and water and methyl acid(30 ∶ 70 ∶ 1),the detection wavelength at 327 nm and the flow rate being 0.5 mL/min.Results A linearity of chlorogenic acid in FP was obtained in the range of 0.16 ? g～ 0.58 ? g,r=0.9996(n=6).The average recovery was 98 % and RSD =1.0% (n=6).Conclusion This method is easy,sensitive and accurate for the determination of chlorogenic acid in FP.
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Using method of orthogonal design we observed that low K+,high Ca2+ & low Mg2+ in perfusate produced the peak incidence of reperf-usion-indaced ventricular fibrillation & introduction of K+ & Ca2+ significantly affected the incidence & pnset of it, The appropriate prop-ortion of K+, ea2+ & Mg2+ in perfusate is the important effecting factor of reperfusion-induced arrhythmias in Langendorff heart of rats.