Tacrolimus strongly inhibits multiple human UDP-glucuronosyltransferase (UGT) isoforms.

The objective of the present study is to clearly evaluate the inhibitory effects of tacrolimus (tacro) on important UGT isoforms in human liver, including determination of inhibition kinetic type and calculation of inhibition kinetic parameters. An in vitro incubation system was used to investigate the inhibitory effect of tacro on UGT isoforms. The recombinant UGT isoforms were used as enzyme source, and a nonspecific substrate 4-methylumbelliferone (4-MU) was utilized as substrate. Among the tested UGT isoforms, UGT1A1, UGT1A3, UGT2B7 and UGT2B15 were strongly inhibited by tacro in a concentration-dependent manner. Dixon and Lineweaver-Burk plots showed that the inhibition of UGT1A1, UGT1A3, and UGT2B7 was all best fit to competitive inhibition type, and the inhibition of UGT2B15 was best fit to noncompetitive type. The inhibition kinetic parameters (Ki) were determined to be 4.7, 1.3, 1.9, and 4.3 microM for UGT1A1, UGT1A3, UGT2B7, and UGT2B15, respectively. Inhibition of these important UGT isoforms in human liver might be an important reason for clinically frequent drug-drug interaction between tacro and other drugs.

Up-regulation of VSIG4 alleviates kidney transplantation-associated acute kidney injury through suppressing inflammation and ROS via regulation of AKT signaling.

Prolonged cold ischemia (CI) is a risk factor for acute kidney injury (AKI) after kidney transplantation (KT). AKI is an abrupt and rapid reduction in renal function due to multi-factors, including inflammation, oxidative stress and apoptosis. V-set immunoglobulin-domain-containing 4 (VSIG4) is a B7 family-related protein and specifically expressed in resting tissue-resident macrophages to mediate various cellular events. In the study, we attempted to explore the effects of VSIG4 on CI/KT-induced AKI in a mouse model. Our results showed that VSIG4 expression was markedly down-regulated in serum of kidney transplant recipients with acute rejection, and in renal tissues of cold ischemia-reperfusion (IR)-operated mice with AKI, which was confirmed in murine macrophages stimulated by oxygen glucose deprivation/reoxygenation (OGD/R). We then found that exogenous VSIG4 markedly ameliorated histological changes in kidney of CI/KT mice by suppressing inflammation and apoptosis through restraining nuclear factor-κB (NF-κB) and Caspase-3 activation, respectively. Oxidative stress and reactive oxygen species (ROS) accumulation in renal tissues were also mitigated by exogenous VSIG4 in CI/KT mice through improving nuclear factor-erythroid 2 related factor 2 (Nrf2) nuclear expression. The inhibitory effects of VSIG4 on inflammation, ROS generation and cell death were confirmed in OGD/R-treated macrophages, which further ameliorated oxidative damage and apoptosis in podocytes. More in vivo and in vitro studies showed that CI/KT- and OGD/R-induced AKI was further accelerated by VSIG4 knockdown. Mechanistically, VSIG4 directly interacted with AKT, and AKT activation was necessary for VSIG4 to govern all these above mentioned cellular processes. Collectively, our findings demonstrated that VSIG4 could mitigate AKI in a CI/KT mouse model, and we identified VSIG4/AKT axis as a promising therapeutic target for the treatment of the disease.

[Clinical observation on long-term surviving patients after combined abdominal organ transplantation].

OBJECTIVE: To summarize the treatment experience of long-term surviving patients after combined abdominal organ transplantation. METHODS: From October 2001 to January 2005, 19 patients received combined abdominal organ transplantation in Nanfang Hospital, including 6 with simultaneous kidney-pancreas transplantation (SKPT), 12 with combined liver-kidney transplantation (CLKT), and 1 with simultaneous liver-pancreas transplantation (SLPT). The periods of follow up were from 6 months to 3 years and 8 months. Summarize primary diseases of the patients, factors which impacted on patients long-term survival rate, and immunological characteristics of combined abdominal organ transplantation. RESULTS: All of 19 transplant cases were performed successfully. Among then, 18 were followed up; 16 survived till now; 2 patients undergoing liver-kidney transplantation were dead, one of which died from myocardial infarction in the 18 months after operation, and one died from cytomegalovirus in infection of lung in 13 months; 1 liver-kidney transplantation patient and 2 pancreas-liver transplantation patients experienced acute rejection once; 2 patients were found nephrotoxicity. Among the 18 patients, 4 patients' survival time were over 3 years, 7 over 2 years, 6 over 1 year, 1 over 10 months. CONCLUSIONS: Combined abdominal organ transplantation is effective for treatment of two abdominal organ failure diseases. Factors which impact on patients long-term surviving include choosing suitable recipient, high quality of donated organ, avoidance of surgical complication, the history of myocardial infarction before operation, immunosuppressive regime and virus infection late after transplantation. Combined abdominal organ transplantation has some different immunological characteristics from single organ transplantation.

[Simultaneous liver-pancreas-duodenum transplantation: one case report and review of the literature].

OBJECTIVE: To investigate the effect of treatment on end-stage liver disease and type-I diabetes mellitus with simultaneous liver-pancreas-duodenum transplantation. METHOD: In September 2003, one patient with chronic hepatitis B, liver cirrhosis, hepatic cellular cancer, and insulin-dependent diabetes received simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantation. Liver and pancreas graft function was monitored after transplantation. RESULTS: The function of pancreas allograft was recovered immediately and the patient became insulin-independence postoperatively. The liver allograft was experienced an acute rejection episode and reversed by intravenous bolus methylprednisolone. The recipient was currently liver disease-free and insulin-free more than 21 months. CONCLUSIONS: The simultaneous liver-pancreas-duodenum transplantation is an effective method in the treatment of end-stage liver disease and type-I diabetes mellitus.

[Role of miR-663 in acute renal graft rejection: an in vitro study].

OBJECTIVE: To compare the serum miR-663 levels in renal transplant patients with and without acute rejection (AR) and explore the role of miR-663 acute renal graft rejection. METHODS: Real time-PCR was used to determine serum miR-663 levels in renal transplant recipients with and without AR. MTT assay and Annexin V-FITC assay were employed to examine the viability and apoptosis of human renal glomerular endothelial cells (HRGEC) treated with a miR-663 mimic or a miR-663 inhibitor, and ELISA was performed to detect the expression of inflammation-related cytokines including IL-6, IFN-γ, CCL-2 and TNF-α in the cells. Transwell assay was used to examine the effect of miR-663 mimic and miR-663 inhibitor on the chemotactic capability of macrophages. RESULTS: Serum miR-663 level was significantly higher in renal transplant recipients with AR than in those without AR. The miR-663 mimic significantly inhibited the viability of HRGECs and increase the cell apoptosis rate, while miR-663 inhibitor suppressed the cell apoptosis. The miR-663 mimic increased the expression levels of inflammation-related cytokines and enhanced the chemotactic capability of macrophages. CONCLUSION: miR-663 might play important roles in acute renal graft rejection and may become a therapeutic target for treating AR.

[Expressions of transforming growth factor-beta1 and collagen IV in renal tissues of patients with chronic allograft nephropathy].

OBJECTIVE: To investigate the expressions of transforming growth factor (TGF)-beta1 and collagen IV in the renal tissues of patients with chronic allograft nephropathy (CAN). METHODS: Immunohistochemical method and computer-assisted image analysis system were used to detect the expressions of TGF-beta1 and collagen IV in the renal tissues of patients with CAN, and the association between TGF-beta1 and collagen IV expressions as well as that between their expressions and the pathological grading of CAN were analyzed. RESULTS: The expressions of TGF-beta1 and collagen IV were significantly higher in the renal tissues of the patients than in normal renal tissues (P<0.001), and the expressions tended to increase with the pathological grades of CAN; TGF-beta1 and collagen IV expressions in both the renal glomeruli and the tubulointerstitium were in patients with CAN positively correlated with normal renal tissues (r=0.943, P<0.001; r=0.910, P<0.001). CONCLUSIONS: Abnormal collagen IV deposition is one of the major factors associated with renal fibrosis in CAN, and TGF-beta1 might play an important role in renal fibrosis in CAN through up-regulation of collagen IV in the renal tissues.

[Perioperative management of multiorgan transplantation].

OBJECTIVE: To summarize the experience with perioperative management of multiorgan transplantation. METHODS: From October 2001 to January 2005, 19 patients received multiorgan transplantation in Nanfang Hospital, including 6 with simultaneous kidney-pancreas transplantation (SKPT), 12 with combined liver-kidney transplantation (CLKT), and 1 with simultaneous liver-pancreas transplantation (SLPT). The surgical techniques, application of immunosuppressants, and complication management were reviewed. RESULTS: All transplantation procedures were performed successfully. The transplantation-related complications included tacrolimus-induced renal toxicosis in 1 (5.3%) case, acute graft rejection in 3 (15.8%) cases, intestinal hemorrhage in 2 (10.5%) cases, intra-abdominal hemorrhage in 1 (5.3%) case, and lung infection in 1 (5.3%) case, all of which were cured after proper treatment. CONCLUSIONS: Donor selection, good quality of the donor organ, proper surgical approaches, adequate use of the-mmunosuppressants, and prevention of complications are essential to the success of multiorgan transplantation.

[Establishment of rat models of simultaneous liver and kidney transplantation].

OBJECTIVE: To establish a simple and reliable rat model of simultaneous liver and kidney transplantation. METHODS: The simultaneous transplantation was performed in healthy male SD rats as the recipients and other SD rats of either gender as the donors. The donor liver and kidney were resected simultaneously and grafted into the recipients whose corresponding organs were previously removed. Anastomosis of the portal vein and the inferior vein cava (IVC) inferior to the kidney between the graft and the recipient was performed by a double cuff method, followed by end-to-side anastomosis of the IVC superior to the liver between the donor and the recipient. The urethra and bile duct were anastomised using a simple inside bracket. RESULTS: The time for blood vessel anastomosis and for recipient operation were reduced, with a success rate of 73.3% in the operations. The function of the grafted liver and kidney remained normal. CONCLUSION: This rat model of simultaneous liver-kidney transplantation is simple and reliable.

[Impact of allograft weight and the recipient's body weight on long-term allograft function].

OBJECTIVE: To investigate the relationship of the renal allograft weight and the recipient's body weight with allograft function after transplantation. METHODS: The correlation of the renal allograft weight, the recipient's body weight, the ratio of the allograft weight to the recipient body weight and the mean serum creatinine (sCr) 3 years after transplantation were measured in 108 kidney recipients. RESULTS: The allograft weight was inversely correlated with the mean sCr 3 years after transplantation (P<0.01). CONCLUSION: The renal allograft weight, recipient's body weight and the ratio of allograft weight to recipient's body weight are important indicators of the long-term allograft function after transplantation, and recipients with greater body weight should receive allografts of higher weight.

Clinical evaluation of abdominal multiorgan transplantation.

OBJECTIVE: To evaluate the clinical effect of abdominal multiorgan transplantation in patients with multiorgan failure. METHODS: Simultaneous kidney-pancreas transplantation (SKPT) with enteric drainage of pancreatic exocrine secretions was performed in 2 patients with type 1 diabetes and end-stage renal disease. A combined liver-kidney transplantation (CLKT) was done in a 66-year-old patient with alcoholic liver cirrhosis and uremia. Simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantation (SLPT) was performed in a patient with hepatitis B, hepatocirrhosis, hepatic cellular cancer, and type 1 diabetes. RESULTS: The function of kidney grafts became normal 5 days postoperatively and insulin-independent after treatment with low dose insulin for 10 days in the 2 SKPT patients. For the CLKT patient, both transplanted organs rapidly achieved normal functions after operation but suffered-acute liver graft rejection on postoperation day 10 and the rejection was controlled after methylprednisolone pulse therapy. In the SLPT patient, insulin was withdrawn 5 days after operation, liver allograft function recovered well. All the patients are alive with stable allograft function after following-up for 29, 26, 9 and 6 months, respectively. CONCLUSIONS: Abdominal multi organ transplantation was effective therapy to patients with multiple organ failure. SLPT can reduce acute pancreas rejection and promote the recovery of liver allograft.

Effect of anti-tumor necrosis factor-alpha monoclonal antibody in alleviating renal ischemia-reperfusion injury.

OBJECTIVE: To investigate the effect of anti-tumor necrosis factor-alpha monoclonal antibody (anti-TNF-alpha mAb) in alleviating renal ischemia-reperfusion injury. METHODS: Fifty normal male Sprague-Dawley rats were randomly divided into 3 groups, namely group A that was subjected to ischemia-reperfusion injury with intravenous administration of anti-TNF-alpha mAb (0.1mg/kg.b.w.) 5 min before reperfusion (treatment group), group B with the same injury followed by saline administration in the same manner (control group), and group C with only anesthetization and leparotomy but not ischemia (sham operation group). Routine assays were performed for testing the levels of blood creatine (Cr), blood urea nitrogen (BUN), plasma TNF-alpha and the cell apoptosis. Ultrastructure of the kidney was also observed. RESULTS: Renal ischemia-reperfusion resulted in significant increase of the levels of Cr, BUN and TNF-alpha in the plasma (P<0.01), but these effects were offset by administration of anti-TNF-alpha mAb (P<0.01). In group B, widespread pathological changes and cell apoptosis were observed in the renal tissue following renal ischemia-reperfusion injury, while similar changes were scarcely visible in group A due to the protective effect of intravenous administration of anti-TNF-alpha mAb 5 min before reperfusion. CONCLUSION: Renal ischemia-reperfusion injury can be alleviated by anti-TNF-alpha mAb treatment.

[Thirteen-year follow-up in 8 cases of penis reconstruction by ilio-inguinal and umbilical-thoracic compound flaps].

OBJECTIVE: To observe the long-term effect of ilio-inguinal and umbilical-thoracic compound flaps in one-stage reconstruction of the penis. METHODS: Eight patients, who received one-stage reconstruction of the penis using ilio-inguinal and umbilical-thoracic compound flaps, were all followed up regularly at 6 months, 1, 3, 7 and 13 years postoperatively. The color, diameter, length, and sense recovery of the organ, along with urodynamics and satisfaction degree of both patients and their wives were recorded. RESULTS: The reconstructed organ retained the original color all through the follow-up, but had shrunk and shortened to some degree within the first 3 years after the operation, a condition that was stabilized afterwards. The sense of the proximal end of the constructed organ began to recover six months after the operation and almost assumed normal sense. The sense of the distal end also recovered, though relatively slowly. Few long-term complications were observed, and the patients and their wives were reasonably satisfied with the reconstructed organs in terms of their shape and function. CONCLUSION: Good long-term effect of one-stage reconstruction of the penis can be achieved by using illio-inguinal and umbilical-thoracic compound flaps, which can be adopted clinically in cases of penis reconstruction.

[Combined liver-kidney transplantation in a senior patient].

OBJECTIVE: To study the surgical techniques, perioperative management, management of infections and graft rejection in patients with combined liver-kidney transplantation (CLKT). METHODS: CLKT was performed in a 66-year-old patient with alcoholic liver cirrhosis and uremia. Lavage in situ with University of Wisconsin (UW) solution of the donor organs and en hoc resection was performed. Orthotopic liver transplantation (OLT) and routine kidney transplantation were respectively carried out. Immunosuppression therapy consisted of tacrolimus (FK506), antithymocyte globulin (ATG), mycophenolate mofetil (CellCept, MMF) and corticosteroid. RESULTS: Both of the transplanted organs rapidly recovered normal functions after operation, and acute rejection of the liver graft occurred on day 10 after operation but was controlled after methylprednisolone pulse therapy. The patient fully recovered and was discharged from hospital on day 29 after operation. CONCLUSIONS: CLKT is effective against both liver and renal function failure. Well-matched HLA tissue typing, proficient surgical skills, adequate application of immunosuppressants and effective management of postoperative complications are crucial for successful CLKT.

[Simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantation in a diabetic patient with end-stage liver disease].

OBJECTIVE: To study the effect of combined transplantation of the liver and the pancreas in diabetic patients with end-stage liver disease, and explore the optimal surgical procedure. METHODS: Simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantations were performed in a patient diagnosed as having chronic hepatitis B, hepatocirrhosis, hepatic cellular cancer, and insulin-dependent diabetes. Immunosuppression therapy utilized prednisone, tacrolimus (FK506), mycophenolate mofetil (MMF), and simulect. The function of the liver graft, serum amylase and lipase, blood glucose, and C-peptide were monitored after transplantation. RESULTS: Insulin was withdrawn at the 6th day after operation, good liver allograft functional recovery was achieved, without such complications as pancreatitis, thrombosis, and localized infections. CONCLUSION: End-stage liver disease with concomitant insulin-dependent diabetes is the indication for combined liver-pancreas transplantations, for which simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantations may constitute the optimal surgical approaches as the primary choice.

[The role of miR-223 in the acute rejection after kidney transplantation].

AIM: To explore the role of miR-223 in the acute rejection after kidney transplantation. METHODS: 33 patients who received kidney transplantation in our hospital within a year were collected and 12 of them appeared acute rejection within 1 month after surgery. In all the patients, the peripheral blood miR-223 level was collected and detected by blind arrangements. Furthermore, PBMCs from healthy volunteers were also collected and stimulated by PHA and then miR-223 level was measured. RESULTS: In the peripheral blood cell, the miR-223 was increased 2 times after acute rejection, as well as 3. 76 times after PHA treatment.Furthermore, using miR-223 predicts AR had a specificity of 90% and sensitivity of 92%. CONCLUSION: The miR-223 may have significant role in the acute rejection of kidney transplantation.

[Effect of CD40 blockade on acute renal graft rejection in rats].

OBJECTIVE: To explore the effect of CD40 blockade in suppressing acute rejection of renal graft in rats. METHODS: With Wistar rats as the donor and male SD rats as the recipients, rat models of acute renal graft rejection was established. The rat models were divided into therapy group and control group, and in the former group, CD40 ligand (CD40L) monoclonal antibody was injected daily for 4 consecutive days starting on the next day following kidney transplantation. On day 5 after the transplantation, the renal graft was harvested for histological examination, and graft rejection was evaluated semiquantitatively. RESULTS: The mean semiquantitative score of the renal graft was 0.63∓0.52 in the therapy group, significantly lower than that of the control group (3.72∓1.48, P<0.05). CONCLUSION: CD40L monoclonal antibody can inhibit acute renal graft in rats.

[Clinical study of pulmonary infection in kidney transplantation recipients taking new immunosuppressant].

OBJECTIVE: To explore the etiopathogenesis, therapy and incidence of pulmonary infection in kidney transplantation recipients taking new immunosuppressant. METHODS: The clinical data from 752 kidney transplant recipients were retrospectively analyzed, who were divided into 3 groups according to the immunosuppressants administered, namely group A (CsA+MMF+Pred, n=226), group B (FK506+MMF+Pred, n=386) and group C (FK506+Rap+Pred, n=140). The incidence and mortality of pulmonary infection were recorded and the analysis of etiopathogenesis, diagnosis and therapy of pulmonary infection were carried out in the 3 groups. RESULTS: Fifty-three patients acquired post-transplant pulmonary infection. The incidence of pulmonary infection was 7.08% (16/226) in group A, 7.25% (28/386) in group B and 6.43% (9/140) in group C. One patient died in group A and 2 in group B. Among the 53 patients, 24 had simple bacterial infection, 9 had cytomegalovirus infection, 1 had mycotic infection, 17 had combined infection, and 2 had unidentified pathogen infection. Of the pathogenic bacteria detected, 68.35% were Gram-negative. CONCLUSION: Gram-negative bacteria are most likely responsible for pulmonary infection after kidney transplantation, which most possibly occurs within 6 months after kidney transplantation. Early diagnosis and early treatment are critical for decreasing the mortality of severe pneumonia and for improving the survival rate of the patients and grafts.

[Application of tacrolimus and cyclosporine A in HBV-carrying renal transplant recipients].

OBJECTIVE: To compare the long-term effect and safety of tacrolimus (FK506) and cyclosporine (CsA) in kidney transplant (KT) recipients carrying hepatitis B Virus(HBV). METHODS: A total of 109 patients with HBV were randomized into FK506 group (52 cases) and CsA group (57 cases) after KT, and a 2-year-long follow-up of the patients was conducted to record the patient and graft survival, incidence of acute graft rejection and postoperative liver function. RESULTS: The 2-year patient/graft survival was 86.0%/73.7% and 94.2%/90.3% in CsA and FK506 groups, respectively (P<0.05), with incidence of acute rejection of 10.5% and 9.6% (P>0.05), and rate of abnormal liver function of 26.3% and 15.4% (P<0.05), respectively. Eight patients (14.4%) in CsA group required a drug conversion but none in FK506 group. The drug conversion resulted in significant reduction of ALT/AST level from 255.13+/-31.38/201.88+/-21.25 U/L to 31.25+/-11.50/25.13+/-9.68 U/L (P<0.01). CONCLUSION: For HBV-carrying renal transplant recipients, FK506 as the primary choice of immunosuppressant can be more effective and safer than CsA.

[Retrospective study of the risk factors of transplant renal artery stenosis].

OBJECTIVE: To investigate the risk factors of transplant renal artery stenosis (TRAS). METHODS: The clinical records of 26 patients undergoing renal transplantation in our hospital between 2000 and 2005 were retrospectively analyzed, whose final diagnosis of TRAS was established on the basis of arteriographic findings. A case-control group of 52 post-renal transplantation patients were sampled by stratified randomization, whose blood pressure and renal graft function were without complications of avascularity or urinary passage. The two groups were matched for the operation time, gender, age, primary diseases, blood type, PRA and HLA matching and use of immunosuppressants. Possible events related to TRAS such as cold ischemia time, acute rejection, delayed graft function and approaches of arterial anastomosis were compared. RESULTS: Fifteen patients (57.7%) with TRAS had a history of acute rejection episode, 7 (26.9%) had delayed graft function, both rates of which were higher than those in the control group (P<0.05). The cold ischemic time and type of arterial anastomosis showed no significant effect on TRAS occurrence (P>0.05). CONCLUSIONS: Post-transplant renal artery stenosis is closely associated with acute rejection and delayed graft function but not with the cold ischemic time or the type of arterial anastomosis.