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Objective To investigate the role and mechanism of spliced X-box binding protein 1 (XBP1s) in the senescence of primary renal tubular epithelial cells induced by hypoxia/reoxygenation (H/R). Methods Primary renal tubular epithelial cells were divided into the normal control group (NC group), H/R group, empty adenovirus negative control group (Ad-shNC group), targeted silencing XBP1s adenovirus group (Ad-shXBP1s group), empty adenovirus+H/R treatment group (Ad-shNC+H/R group) and targeted silencing XBP1s adenovirus+H/R treatment group (Ad-shXBP1s +H/R group), respectively. The expression levels of XBP1s in the NC, H/R, Ad-shNC and Ad-shXBP1s groups were measured. The number of cells stained with β-galactosidase, the expression levels of cell aging markers including p53, p21 and γH2AX, and the levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined in the Ad-shNC, Ad-shNC+H/R and Ad-shXBP1s+H/R groups. Chromatin immunoprecipitation was employed to verify Sirtuin 3 (Sirt3) of XBP1s transcription regulation, and the expression levels of Sirt3 and downstream SOD2 after down-regulation of XBP1s were detected. Mitochondrial reactive oxygen species (mtROS) were detected by flow cytometry. Results Compared with the NC group, the expression level of XBP1s was up-regulated in the H/R group. Compared with the Ad-shNC group, the expression level of XBP1s was down-regulated in the Ad-shXBP1s group (both P<0.001). Compared with the Ad-shNC group, the number of cells stained with β-galactosidase was increased, the expression levels of p53, p21 and γH2AX were up-regulated, the levels of ROS, MDA and mtROS were increased, the SOD activity was decreased, the expression level of Sirt3 was down-regulated, and the ratio of Ac-SOD2/SOD2 was increased in the Ad-shNC+H/R group. Compared with the Ad-shNC+H/R group, the number of cells stained with β-galactosidase was decreased, the expression levels of p53, p21 and γH2AX were down-regulated, the levels of ROS, MDA and mtROS were decreased, the SOD activity was increased, the expression level of Sirt3 was up-regulated and the ratio of Ac-SOD2/SOD2 was decreased in the Ad-shXBP1s+H/R group (all P<0.05). Conclusions Down-regulation of XBP1s may ameliorate the senescence of primary renal tubular epithelial cells induced by H/R, which probably plays a role through the Sirt3/SOD2/mtROS signaling pathway.
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Objective:To explore the protective effect of methyl eugenol (Me) on islet ischemia/reperfusion (I/R) injury and elucidate its underlying mechanism.Methods:The islets were isolated and purified from 6-8 week male BALB/c mice and divided into four groups of normal control (normal culture without any treatment), hypoxia/reoxygenation (H/R treatment), H/R+ dimethyl sulfoxide (DMSO dosing plus H/R treatment) and H/R+ Me (Me dosing plus H/R). Viability of islet cells in each group was detected by acridine orange (AO)/propidium iodide (PI) double stain.Function of islet cells (insulin secretion) was measured by enzyme-linked immunosorbent assay (ELISA). Murine islet β Min6 cells were selected for detecting the effect of Me on the proliferative activity of normal cultured and H/R treated islet cells under different concentration gradients by CCK8.Then Min6 cells were divided into four groups of normal, H/R, H/R+ DMSO and H/R+ Me.The definition of group was the same as that of primary murine islets.Flow cytometry and Hoechst 33342 nuclear stain were utilized for detecting cell apoptotic rate in each group.The protein expressions of p-JNK, p-p38, JNK, p38, Bcl-2 and Bax were detected by Western blot.And the data were processed by one-way ANOVA or t test.Results:The proportion of dead islet cells in H/R group was (29.47±2.65)% and it was significantly lower than that in normal group (7.63±1.53)%.And the inter-group differences were statistically significant ( P<0.001). The proportion of dead islet cells was (20.63±3.07)% in H/R+ Me group.It was higher than that in H/R group (29.47±2.65)% and in H/R+ DMSO group (30.13±1.50)% and inter-group difference was statistically significant ( P<0.05 & P<0.01). Under the stimulation of high glucose, the insulin secretion level of islet in H/R+ Me group was (1.76+ 0.08) mg/L, which was higher than that in H/R group and H/R+ DMSD group(1.24±0.14)mg/L and(1.27±0.05)mg/L, and the difference was statistically significant[(1.76±0.08) vs. (1.24±0.14) mg/L; (1.76±0.08) vs.(1.27±0.05) mg/L, P<0.01]. There was no significant effect on cell viability after Me dosing within a certain concentration range (0-40 μmol/L). After Me dosing (5 μmol/L), cell viability of H/R-treated Min6 cells was significantly higher than that without Me.And the difference was statistically significant[(1.19±0.03) vs.(1.00±0), P<0.01]. As compared with H/R and H/R+ DMSO groups, overall apoptotic rate declined in H/R+ Me group (Hoechst 33342 stain: 14.50%±1.05% vs. 23.30%±1.18%, 14.50%±1.05% vs. 22.77%±1.75%, P<0.001; Flow cytometry: 4.36%±0.54% vs. 21.44%±1.02%, 4.36%±0.54% vs. 21.68%±3.06%, P<0.01). The expressions of p-JNK and p-p38 were down-regulated (p-JNK: 0.77±0.06 vs. 1.03±0.05, 0.77±0.06 vs.0.93±0.04, P<0.001; p-p38: 0.80±0.05 vs. 1.01±0.08; 0.80±0.05 vs. 1.00±0.05, P<0.05) while Bcl-2/Bax ratio rose (1.62±0.13 vs. 0.72±0.10, 1.62±0.13 vs. 0.74±0.13, P<0.01). Conclusions:Me can improve the viability and function of islets and suppress the apoptosis of Min6 cells after H/R.The mechanism is correlated with JNK and p38 MAPK signaling pathways.
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Objective:To explore the safety and feasibility of optimized pathological evaluation system of donor's kidney and modified surgery during adult dual kidney transplantation(DKT)and evaluate its effectiveness to provide more alternative protocols for kidney transplantation from extended criteria donors.Methods:DKT was performed in 10 recipients using the same protocol from June 2019 to May 2021.And retrospective reviewing was performed for clinical data, including characteristics of donors and recipients, optimized pathological evaluation system, modified surgery, treatment regimens, complications and follow-ups.Results:There were 8 male and 2 female donors with an age of(57.9±12.8)years and BMI(24.1±4.1)kg/m 2.The percentage of DCD was 70% and DBD 30%.The serum creatinine before procurement was 107.6(93.3-163.5)μmol/l.Zero-point puncture biopsy was performed for both kidneys and optimized pathological evaluation system was implemented(Banff criteria & Remuzzi score). The pathological results indicated that glomerular sclerosis for left and right kidneys were 2.0(1.5-2.0)and 1.5(1.0-2.0). And Remuzzi score for left and right kidneys were(4.4±1.2)and(3.6±1.5)points respectively.All recipients were male with an age of(43.1±9.0)years and BMI(22.2±1.9)kg/m 2.All PRAs were negative pre-operation.Modified surgery was performed in all recipients(two kidneys were implanted outside iliac vessels without patch and artery of superior kidney was anastomosed to internal iliac artery). Operative duration was(195±54.3)min and serum creatinine before discharge 125.0(102.0-199.0)μmol/L.Renal dynamic scintigraphy indicated that glomerular filtration rate was(30.0±8.2)ml/min for left kidney and(29.2±13.9)ml/min for right kidney.MRA results indicated that morphologies of renal arteries and veins were regular.The time between operation and discharge was(22.4±4.7)days.Compared with SKT, serum creatinine before discharge of DKT was lower and DGF incidence of DKT was higher without statistical significance.The time between operation and discharge was longer for DKT than that for SKT( P<0.05). The complications consisted of 20% donor derived infection(DDI)and 50% DGF.And there was no surgical complication associated with vessels and ureter.Renal function remained stable during 6-month follow-ups. Conclusions:Optimized pathological evaluation system of donor's kidney and modified surgery during adult dual kidney transplantation are both safe and feasible.The postoperative function of transplanted dual kidney is successfully restored.However, long-term follow-ups are required for evaluating its effectiveness.
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Objective@#To analyze the clinical characteristics of one living-related kidney transplant recipient infected with 2019 coronavirus disease (COVID-19) .@*Method@#The clinical diagnosis and treatment of one relative renal transplant recipient after the occurrence of COVID-19 were analyzed retrospectively, including the course of onset, clinical manifestations, blood routine test, renal function, lung CT scan, nucleic acid detection, outpatient and inpatient therapies and outcomes.@*Result@#The case was diagnosed as COVID-19 (severe type) with influenza A virus infection. The clinical symptoms were gradually relieved and the lung lesions were absorbed through the treatment of reduce and stop taking immunosuppressant, antiviral therapy of abidol/oseltamivir, prevention of bacterial infection, hormone anti-inflammatory, oxygen inhalation, nutritional support and adequate rest.@*Conclusion@#This case present typical characteristics of COVID-19 in epidemiological investigation, clinical manifestation, examination, pulmonary imaging and etiology. After comprehensive treatment including reduce and stop immunosuppressive therapy, clinical cure was achieved. The long-term effect of COVID-19 on this immunosuppressive patient remains follow-up.
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Objective:To explore the clinical characteristics of one living-related kidney transplant recipient infected with 2019 coronavirus disease(COVID-19).Methods:The clinical diagnosis and treatment of one living-related kidney transplant recipient after the occurrence of COVID-19 were analyzed retrospectively. Course of onset, clinical manifestations, laboratory and image enamination, outpatient and inpatient therapies and outcomes.Results:The renal transplant recipient was diagnosed as COVID-19(severe) with influenza A virus infection based upon epidemiological survey, clinical manifestations, laboratory examinations, imaging findings and etiological tests. The clinical symptoms were gradually relieved and lung lesions became absorbed after tapering and withdrawing immunosuppressants, antiviral therapy of abidol/oseltamivir, antibiotic therapy, hormonal anti-inflammation, oxygen inhalation, nutritional supports and adequate rest.Conclusions:Living-related kidney transplant recipients have specific immunosuppressive states.The long-term effect of covid-19 on recipients should be determined through long-term follow-ups.
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Objective:To summarize the transplant outcomes of pediatric kidney transplantation at a single center and discuss probable measures of improving the outcomes.Methods:A total of 111 pediatric renal transplantation were performed from September 2002 to September 2019. They were divided into adult-donor group ( n=41) and pediatric-donor group ( n=70). Adult-donor group consisted of two subgroups based upon donor sources: living-donor group ( n=19) and deceased-donor group ( n=22). Pediatric-donor group consisted of two subgroups based upon surgical types: single kidney group ( n=48) and en bloc kidney group ( n=22). Clinical data and outcomes of grafts and recipients were retrospectively analyzed. Results:The average age of recipients was (15.6±1.9) years in adult-donor group. None developed delayed graft function (DGF) in living-donor group whereas 6 patients (27.3%) had DGF in deceased-donor group ( P<0.05). During a follow-up period of 22-181 months, 1-year and 5-year graft survivals were 100% vs 94.1% and 93.8% vs 94.1% in living-donor and deceased-donor groups respectively. There were no statistical differences. In pediatric-donor group, the age of donors was significantly lower in en bloc subgroup than that in single kidney subgroup (median: 0.5 vs 6 months, P<0.05). The age of recipients was similar between two subgroups: (9.5±5.3) years in single kidney group vs. 11.5± 1.8 years in en bloc kidney group. In addition, 7 cases of single kidney were transplanted for infant recipients aged under 1 year. Vascular thrombosis occurred in 3 patients (6.3%) of single kidney group, less than that in 5 patients (22.7%) of en bloc kidney group ( P=0.06). During a follow-up period of 4-54 months, 1-year and 2-year graft survivals were 85% and 80% in single kidney group whereas 75% and 70% in en bloc kidney group. However, there was no statistically significant difference. One-year survival was 98% in single kidney group and 95% in en bloc kidney group. Conclusions:For elder pediatric recipients, excellent kidney transplant outcomes may be achieved with grafts from adult donors. For pediatric kidney recipients, transplant outcomes can be further improved with careful assessments and cautious usage of small grafts, particularly those form neonatal donors.
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Objective To investigate the clinical efficacy of vena cava-atrium anastomosis liver transplantation (VCAALT) for Budd-Chiari syndrome (BCS).Methods The retrospective descriptive study was conducted.The clinicopathological data of 18 BCS patients who underwent VCAALT in the Zhongnan Hospital of Wuhan University (6 cases),the Third Xiangya Hospital of Central South University (8 cases) and Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology (4 cases) from May 1996 to December 2012 were collected.All the 18 patients were males,aged from 29 to 61 years,with an average age of 42 years.According to characteristics and invasion extent of hepatic vein and vena cava after preoperative examinations,patients were performed different surgical procedures of VCAALT,including bridge piggyback liver transplantation (BPBLT),hanging atrium liver transplantation (HALT) and cava vena resection bridge liver transplantation (CVRBLT).Observation indicators:(1) surgical and postoperative situations;(2) typical case analysis;(3) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect patients' survival up to December 2018.Measurement data with normal distribution were represented as Mean±SD and measurement data with skewed distribution were described as M (range).Results (1) Surgical and postoperative situations:of 18 patients,11 underwent BPBLT,3 underwent HALT,4 underwent CVRBLT.The operation time and volume of intraoperative blood loss were (6.0± 1.3)hours and (1 264±435)mL.One patient died of bilateral pulmonary diffuse inflammation and sepsis due to severe infection.The duration of postoperative hospital stay was (18±5) days.(2) Typical case analysis:one 47-year-old male BCS patient was detected retrohepatic vena cava plaques and thrombus and hepatic venous thrombus by exploratory laparotomy,and underwent BPBLT.A 43-year-old male BCS patient was detected hepatic and retrohepatic vena cava plaques,thrombus,concomitant cavernous transformation,and underwent HALT.A 32-year-old male BCS patient was detected plaques and thrombus with red thrombus in the hepatic vein,from right renal vein to right atrium,and underwent CVRBLT.All the 3 patients underwent VCAALT successfully with a satisfactory recovery.(3) Followup situations:18 patients were followed up for 3.0-60.0 months,with a median time of 51.7 months.During the follow-up,3 patients died of acute rejection,biliary complications and chronic graft dysfunction at 1,3,5 years postoperatively.The 1-,3-,5-year survival rates were 16/18,15/18,14/18,respectively.Conclusion Different surgical procedures of VCAALT for BCS are selected according to different situations of patients,which are safe and feasible with a satisfactory efficacy and beneficial to long-term survival of patients.
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Objective To explore the influencing factors and treatment strategies of long-term survival after simultaneous pancreas and kidney transplantation (SPK ) .Methods One case of long-term post-SPK survival was reviewed and its influencing factors were analyzed along with the relevant literature .Results At 10 years post-SPK ,the patient lost transplanted kidney due to rejection and underwent secondary kidney transplantation . The transplanted pancreas functioned well and has survived for more than 18 years .Conclusions Strict preoperative screening ,adopting mature surgical approaches ,aggressive managements of various perioperative complications ,strengthening of health education of recipients ,improving of compliance and long-term regular follow-ups are conducive for enhancing long-term survival of recipients and grafts of SPK .
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Objective To investigate the clinical outcome of single kidney transplantation from pediatric donors and to explore the application criteria.Methods The clinical data of 14 recipients undergoing renal transplantation from October 2006 to October 2016 were retrospectively analyzed.All the recipients received primary kidney transplantation from pediatric donor and the renal artery was anastomosed with external iliac artery.Based on the length of the kidney donor,the recipients were divided into two groups as group A (length beyond 6 cm) and group B (length 5-6 cm).The clinical effect and complications of the 14 recipients,the survival of the recipients and grafts,the recovery of renal function,the change of the renal length and the postoperative complications were compared between the two groups.Results The renal length in group A (n =7) was (7.5 ± 1.2) cm,and (5.7 ± 0.1) cm in group B (n =7).During the follow up period,all renal grafts and recipients survived.No significant difference was observed between two groups in renal graft function evaluated by serum creatinine and estimated glomerular filtration rate (eGFR) at 7th day,14th day,1st,2nd,3rd and 6th month postoperatively,P＞0.05.The length of transplanted kidney increased after operation in both groups,with results of 9.9 ± 0.6 cm in group A and 10.4 ± 1.5 cm in group B (P＞0.05),respectively,at 2nd month post-transplantation.Delayed graft function (DGF) occurred in 2 cases of group A and 1 case of group B.Seven cases developed proteinuria (50.0％),including 2 cases in group A (28.6％) and 5 cases in group B (71.4％).Four cases suffered hematuria (28.6％),including 2 cases in group A (28.6％) and 2 cases in group B (28.6％).One recipient in group B suffered acute rejection.No vascular embolization,urine leakage,pulmonary infection and other complications were observed in all the recipients.Conclusion The length beyond 5 cm is acceptable for single pediatric kidney donor for adult recipients with a promising clinical outcome in short-term.However,the high incidence of proteinuria and hematuria remains obstacle,and the long-term outcome needs further exploration.
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Objective To investigate the value of different biopsy methods for quality evaluation of the donated kidney organ after citizen death.Methods Six cases (6 pairs) of discard donor kidneys were collected from October 2016 to May 2017,respectively,and grouped by wedge biopsy and core needle biopsy.After being fixed and processed for routine paraffin embedding and hematoxylin-eosin (H&E) staining,the specimens were evaluated by the qualified rate of sample,the number of glomerulus,sclerotic glomerulus and small arteries.Results The comparison of two different biopsy methods showed that the number of samples obtained by wedge biopsy was 30.There were 29 samples which were qualified and the qualification rate was 96.7％.The number of samples obtained by core needle biopsy was 30,and only 21 samples were qualified and the qualification rate was 70％.In the wedge biopsy samples,the average number of glomeruli was 22.1 and 6.9 of them were sclerotic glomeruli.The ratio of sclerotic glomeruli was 31.3％.The average number of glomeruli in core needle biopsy samples was 9.5 and 2.1 of them were sclerotic glomeruli.The ratio of sclerotic glomeruli was 22.1 ％.The average number of arteries in wedge biopsy samples was 5.4,and that in core needle biopsy samples was 3.9.The results indicated that the qualification rate of wedge biopsy was significantly higher than that of core needle biopsy (P＜0.01).The number of glomeruli,sclerotic glomeruli and small arteries in wedge biopsy samples was significantly greater than than in core needle biopsy (P＜0.05).Conclusion Wedge biopsy was superior to core needle biopsy for the quality evaluation of specimens and identifying clinically significantly histopathological findings.Thus it is potential for wedge biopsy to become the main method in pre-implantation histopathological evaluation.
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Objective To analyze the safety of renal transplant from donors with primary central nervous system (CNS) tumors.Methods We retrospectively analyzed the clinical data of 33 donors with primary CNS tumors and the 63 corresponding renal recipients between January 2013 and December 2016 in Tongji Hospital.Results The mean period from diagnosis as primary CNS tumor to donation was about (21.8± 46.4) months (range:0.5 to 192.0 months).The pathological classification of these tumors included gliomas,meningioma,medulloblastoma,etc.Besides,there were 10 donors with high-grade CNS malignancies.Eleven donors have ever been through at least one of the four treatments (craniotomy,V-P/V-A shunt,radiotherapy and chemotherapy),14 donors have undergone none,and the clinical data of rest were unavailable.All the 63 recipients got well renal function after transplant.During an average follow-up of (15.9 ± 8.2) months (range:2.7 to 35.5 months),one recipient got donor-derived rhabdoid tumor 4 months posttransplant,underwent comprehensive treatments,including allograft nephrectomy,radiotherapy,chemotherpy and returned to hemodialysis,while the 62 cases got no donor-derived tumors.Conclusion Tumor transmission of renal allograft from donors with primary CNS tumors is inevitable but with low risk,which means this kind of donors can be used with careful assessment,full informed consent and good balance between wait-list death and tumor transmission.
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Objective To investigate the feasibility and safety of the single kidney transplantation from pediatric donors to adult recipients.Methods From May 2013 to January 2017,a total of 50 single kidney transplants from pediatric donation after citizen death (DCD) donors of age between 3 to 12 years to adult recipients were performed and the data were summarized.Results The average age of donors was 6.4 ± 2.5 years with an average donor weight of 19.1 ± 5.9 kg,and the average kidney length was 6.3 ± 0.6 cm.For the 50 adult recipients,the average age was 38.5 ± 12.1 years,the average body weight was 56.1 ± 13.1 kg,and the number of female patients was 26 (52％).All except 3 of these patients were transplanted for the first time.Delayed graft function (DGF) was observed in 15 patients (30％).The average value of eGFR among all the patients was rapidly increased in the first 3 months after transplantation and then steadily increased to (82.3 ± 13.4) mL· min-1·1.73 m-2 at 1 st year,followed by (83.8 ± 22.5) mL· min-1·1.73 m-2 at 2nd year.Four renal grafts developed acute rejection (8％),and 3 of them were successfully reversed by the treatment.Pulmonary infection occurred in 4 recipients,and 2 died.During a follow-up period of 19 months,uncensored grafts survival was 94％,and patients survival was 96％.Conclusion Excellent intermediate-term transplant outcome can be achieved by using single kidneys from pediatric donors elder than 3 years,which may shorten the waiting time in adult recipients and alleviate the contradictions in the absence of suitable pediatric recipients.
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Objective To explore the mechanism of hepatic autophagy inhibition induced by ischemia-reperfusion injury in the aging liver.Methods The healthy male Lewis rats aged 3 months (3M) and 24 months (24M) were selected,and then were randomly divided into 3M IRI group,3M sham operation group,24M IRI group,24M sham operation group.In the experimental group,noninvasive vascular clamp was used to clamp the left and middle hepatic lobes (about 70o％ hepatic ischemia).The liver was subjected to ischemia at 37 0.5℃ for 30min and reperfusion for 6h.The hepatic duodenal ligament was dissected only by sham operation.The serum levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured at 6 h after operation.Liver tissues of each group were examined by liver pathology and the number of autophagosome of LC3B in liver tissue of each group was observed under confocal microscopy.The changes of autophagyrelated protein were analyzed by Western blotting.Results The levels of serum ALT and AST in 24M IRI group were significantly higher than those in 3M IRI group,the difference was statistically significant (P＜0.05).Pathological analysis showed that 3M IRI group showed spotty necrosis,the 24M IRI group showed massive necrosis and the infiltration of the inflammatory cells;Confocal microscopy showed that the number of autophagosome in the liver tissue of the 24M sham group was slightly lower than that of the 3M sham operation group and the number of autophagosome in the 24M IRI group was significantly lower than that in the 3M IRI group (P＜0.05).The levels of autophagyrelated proteins (Beclin1 and ATG4B protein) in 24M IRI group were significantly down-regulated compare to 3M IRI group (P＜0.05).Conclusion The ischemia-reperfusion injury of liver in aged rats inhibits autophagy,and its mechanism may be related to the decrease of autophagy-related protein level in hepatic ischemia-reperfusion injury.
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Objective To investigate the clinical characteristics and strategies of early stage antibody-mediated rejection after renal transplantation.Method The clinical data of early stage AMR of 3 cases of renal transplantation,and 1 case of pancreas transplantation after renal transplantation were retrospectively analyzed.(1) The case 1 was diagnosed as having early severe acute AMR.Serum creatinine was increased,urine volume rapidly reduced,the blood flow of transplanted kidney reduced on the postoperative day 8;the positive rate of panel reactive antibody (PRA) class Ⅰ and Ⅱ was 74.6％,and 2.7％ respectively on the postoperative day 12.Biopsy showed widely ischemia and local bleeding in transplanted kidney and DSA showed anti-B62 mean fluorescence intensity (MFI) increased to 6800 on the postoperative day 14.(2) The case 2 was diagnosed as having early mild acute AMR.The positive rate of PRA class [and Ⅱ was 65.6％ and 78.9％ respectively.DSA Ⅰ was positive,anti A11 MFI was 3059,and DSA Ⅱ was negative on the postoperative day 13.Biopsy showed mild ischemia reperfusion injury in transplanted kidney on the postoperative day 21.(3) The case 3 was diagnosed as having early severe chronic AMR,and the recipient received pancreas transplantation 1 year after kidney transplantation.Eight months after pancreas transplantation,DSA for pancreas donor was detectable,anti A2 MFI was 7514,anti B46 MFI was 3 159 and anti DQ7 MFI was 1 503.(4) The case 4 was diagnosed as having early mixed rejection.Serum creatinine was elevated on the postoperative day 8;PRA testing showed that the positive rate of class Ⅰ and Ⅱ was 3％ and 70％ respectively,DSA was positive,and anti DR16 MFI was 15 170 on the postoperative day 14;transplanted kidney biopsy showed acute mixed rejection on the postoperative day 16.Result Case 1 and case 3 were not diagnosed and treated in time and graft loss developed.Case 2 and case 4 were functionally recovered after combined treatment of plasmapheresis,IVIG and bortezomib.Conclusion Diagnosis of antibody-mediated rejection is based on transplant graft dysfunction,positive DSA and graft biopsy.Early diagnosis,early treatment and combined therapy can improve the curative rate of AMR.
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Objective To investigate the effect of down-regulated Blimp1 gene expression on differenetiation of bone marrow cells into dendritic cells (DCs).Methods Blimp1-shRNA was constructed and then loaded into lentivirus vector as lenti-blimp1-shRNA.Bone marrow cells from Balb/c mice were induced differentiation to DCs in an 8-day cell culture system with GM-CSF/IL-4 incubation and LPS stimulation at day 7.The cells were divided into groups as empty control (no treatment),lenti-control (transfected by lentivirus empty vector at day 1),and lenti-Blimp (transfected by lenti-blimp1-shRNA at day 1).The transfection efficiency was evaluated by GFP fluorescence for one week.The morphology and growth curve were analyzed.Real-time PT-PCR and Western blotting were used to evaluate mRNA and protein expression of Blimp1.At day 8,CD11 c and CD86/MHC-Ⅱ were quatitified using flow cytometry.Results GFP fluorescence emerged 3 days after transfection and was continuously expressed.Classic DC morphology was shown in no treatment cells,while damaged morphology presented in the cells with lentivirus transfection.The empty control cells proliferated from day 3,peaked as (2.45 ± 0.26) 106/well at day 4,and kept at (2.27 ± 0.19) 106/ well at day 8,The cells receiving lentivirus presented (1.69 ± 0.39) 106/well.The expression of Blimp1 mRNA and protein in the lenti-Blimp1 group was 76％/1％ and 1.0％/74.0％ of the empty control group.At day 8,CD11c,CD86 and MHC-Ⅱ expression in the empty control group was (69.2 ±5.0)％,(51.1± 4.9) ％ and (56.3 ± 7.3) ％,while (68.6±5.9)％,(49.5±4.3)％ and (69.4±4.5)％ in the lenti-control group,and (72.8 ± 5.5)％,(50.2 ± 6.0)％ and (46.5 ± 5.7)％ in the lenti Blimp1 group.Conclusion Lentivirus-mediated Blimp1-shRNA gene therapy modulates blimp1 expression of DC precursors.Down-regulation of Blimp1 fails to interrupt the differentiation of DCs but inhibits the maturation.
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Objective To determine the long-term results after combined pancreas-kidney transplantation at a single-center institution.Methods Fifty-three consecutive patients with insulin-dependent diabetes mellitus and end-stage nephropathy were followed up for more than three years after combined pancreas-kidney transplantation. Immunosuppressive protocol consisted of tacrolimus ( TAC ),mycophenolate mofetil (MMF),and steroids,and antithymocyte globulin or anti-CD25 receptor mAb.The impact of different risk factors was analyzed on long term patient and graft survival.Results The 3-,5- and 8-year survival rate in recipients was 90.1％,89.1 ％ and 80.0％,respectively.The 3-,5- and 8-year survival rate of pancreas grafts was 84.9％,84.8％ and 60.0％,and that of kidney grafts was 83.0％,82.6％ and 53.3％,respectively.Principal causes of death were Infection (n =4),renal failure (n =2),cardiovascular events (n =1 ),and cerebrovascular accident (n =1 ).Graft failure for the pancreas was caused by death with a functioning graft (n =6),rejection (n =2),thrombosis (n =1 ) and pancreatitis (n =1 ).Graft failure for the kidney was due to rejection (n =9),and death with a functioning graft (n =9).Conclusion This series representing the largest experience with long-term follow up in China confirms an excellent long-term survival.Infection,rejection and surgical complication were the major risk factors leading to deaths and graft loss.
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Objective To investigate the effects of T. gondii soluble tachyzoite antigen (STAgs) on the survival time of mouse heart allograft and the possible mechanism. Methods The STAgs were prepared by pulverizing T. gondii tachyzoite with ultrasound on ice. Cervical heterotopic heart transplantations were done by using Balb/c mice as donors, and C57BL/6 mice as recipients.The recipients were classified randomly into three groups: syngeneic group, acute rejection group and STAgs-treated group. The recipients in acute rejection group and STAgs-treated group were injected subcutaneously with 0. 1 ml PBS and 0. 1 ml (5 μg) STAgs at the 4th day before transplantation respectively, and those in syngeneic group were not subjected to any treatment. The grafts were observed daily by cervical palpation, and the total cessation of cardiac contraction was defined as the endpoint. The heart allografts were harvested at the 7th day after transplantation for pathological examination and immunohistochemical staining for CD4+ T, CD8+ T. Results The recipients in syngeneic group were all alive at the 100th day after transplantation. The average survival time in acute rejection group and STAgs-treated group was (6.7± 0.5) days and (70.8± 3.5) days,respectively (P＜0.05). HE staining showed that the rejection on the 7th day after transplantation in syngeneic group, acute rejection group and STAgs-treated group was fallen into 0 degree, Ⅲ-Ⅳ degree and 0- Ⅰ degree, respectively. Immunohistochemical staining revealed that the CD4+ T and CD8+T were markedly down-regulated in STAgs-treated group as compared with those in acute rejection group. Conclusion T. gondii STAgs can significantly prolong the survival time of mouse heart allograft and inhibit the rejection probably by changing the ratio of TH1/TH2, or inhibiting the effect of dendritic cells by inducing the lipoxin A4.
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Objective To analyze the curative effect of simultaneous liver and kidney transplantation (SLKT) for patients with end-stage liver and kidney diseases and liver cirrhosis patients with hepatorenal syndrome.Methods All SLKTs (n=21) performed at our center from January 1999 to December 2010 were reviewed and SLKT outcomes were compared with those of kidney transplantation (KT) (n=609) and liver transplantation (LT) (n=133) performed during the same period.Results There were 3 deaths due to infection 2 weeks, 6 months and 5 years respectively after operation. One patient died due to multiple organ dysfunction syndrome 2 weeks after operation. One patient was dead 5 years after operation because of rejection. MELD level between SLKT and LT had no significant difference, but serum creatinine in SLKT group was significantly higher than in LT group (516.0±329.9 vs 111.4±138.1 mmol/L, P<0.01). The 1-year acute kidney rejection rate and rate of delayed graft function (DGF) of the kidney had no significant difference between SLKT group (0 vs 9.5 %) and KT group (6 % vs 17.3 %). There was no significant difference in one-, 3- and 5-year patient survival rate between SLKT group (87.7 %, 67.8 % and 67.8 %) and LT group (84.2 %, 73.5 % and 69.4 %).Conclusion SLKT is a safe and effective treatment for end-stage liver and kidney diseases.
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Objective To evaluate the relevant causes of neurologic complications following liver transplantation.Methods 155 adult patients (131 males, 24 females) who received liver transplantation for the first time at Tongji Hospital between January 2005 and September 2009 were identified.Case notes were reviewed and demographic data, details of the liver disease, neurologic complications, MELD score and discharge information were recorded.Results Neurologic complications occurred following 36 transplants (23.2 %), The complications included mental symptoms in 15 cases (41.7 %), disorder of consciousness and action in 9 cases (25 %), and coma in 12 cases (33.3 %).Twelve percent patients with liver cancer experienced a neurologic complication, which was lower than for other transplant indications, like acute and chronic hepatic failure because of HBV infection (33.3 %, P＜0.01), inborn/metabolic disease (40 %, P＜0.05), and HCV Infection (25 %, P = 0.36).Patients who experienced a neurologic problem had significantly higher MELD score (for non-cancer patients:22.93 ± 8.21; for cancer patients:17 ± 5.4) than the other Patients (for non-cancer patients:18.33 + 8.47, P＜0.05; for cancer patients:13 ±3.4, P＜0.01).The rate of infection (36.1 %) and mortality (30.5 %) were significantly higher in patients with neurologic complications (P＜0.01).The levels of ALT, TBil, ALB, PT and the concentrations of serum sodium and chlorine had no impact on neurologic complications.Conclusion Neurologic complications are common in liver transplant recipients.These complications are related to primary disease and liver function before the operation, and increase the rate of infection and mortality.
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Objective To prepare recombinant adenovirus pAd-gal-9 containing murine galectin-9 and explore galectin-9's pro-apoptotic effect on T lymphocytes. Methods The recombinant adenovirus plas-mid pAd/CMV/V5-DEST-gal-9 was prepared by conventional molecular cloning and LR reaction. The pAd/ CMV/V5-DEST-gal-9 linearlized by Pac I was transfected into 293A cells with Lipofectin 2000. Eight days after transfection, the 293A cells were subjected to freeze/thraw circle for three times and the supernatant was collected after centrifugation. Higer titer pAd-gal-9 was produced by large-scale infection of 293A cells with the supernatant containing pAd-gal-9. The supernatant was condensed to get purified pAd-gal-9 by CsCl density gradient centrifugation. After titer determination with gradient dilution of harvested pAd-gal-9 infec-tion in 293A-seeded 96-wells, pAd-gal-9 was used to infect the CHO cell line. Immunohistological assay, Western blot and flow cytometry were employed to ascertain the subcellular location expression of galectin-9. We added solid-phase transgenic CHO cells or freshly-cultured supernatant to medium containing activated T cells to detect the pro-apoptotic effect of galectin-9. Results The pAd-gal-9 was prepared successful. Im-munohistochemical staining of CHO infected with pAd-gal-9 confirmed that galectin-9 was expressed in the cytosol. Intercellular staining indicated that mean fluorescence intensity of galectin-9 was significantly higher in pAd-gal-9-infected CHO group than control group. Supernatant from pAd-gal-9-infected CHO promoted the apoptosis of T cells. The percent of apoptotic T cells was higher than the Tim-3 positive T cells. Conclu-sion CHO infected with pAd-gal-9 can secret galectin-9 to promote the apoptosis of activated T cells via Tim-3-independent mechanisms.