Dual-specificity phosphatase 26 protects against kidney injury caused by ischaemia-reperfusion through restraint of TAK1-JNK/p38-mediated apoptosis and inflammation of renal tubular epithelial cells.

Dual-specificity phosphatase 26 (DUSP26) acts as a pivotal player in the transduction of signalling cascades with its dephosphorylating activity. Currently, DUSP26 attracts extensive attention due to its particular function in several pathological conditions. However, whether DUSP26 plays a role in kidney ischaemia-reperfusion (IR) injury is unknown. Aims of the current work were to explore the relevance of DUSP26 in kidney IR damage. DUSP26 levels were found to be decreased in renal tubular epithelial cells following hypoxia-reoxygenation (HR) and kidney samples subjected to IR treatments. DUSP26-overexpressed renal tubular epithelial cells exhibited protection against HR-caused apoptosis and inflammation, while DUSP26-depleted renal tubular epithelial cells were more sensitive to HR damage. Upregulation of DUSP26 in rat kidneys by infecting adenovirus expressing DUSP26 markedly ameliorated kidney injury caused by IR, while also effectively reducing apoptosis and inflammation. The mechanistic studies showed that the activation of transforming growth factor-ß-activated kinase 1 (TAK1)-JNK/p38 MAPK, contributing to kidney injury under HR or IR conditions, was restrained by increasing DUSP26 expression. Pharmacological restraint of TAK1 markedly diminished DUSP26-depletion-exacebated effects on JNK/p38 activation and HR injury of renal tubular cells. The work reported a renal-protective function of DUSP26, which protects against IR-related kidney damage via the intervention effects on the TAK1-JNK/p38 axis. The findings laid a foundation for understanding the molecular pathogenesis of kidney IR injury and provide a prospective target for treating this condition.

Development and validation of a novel nomogram model for predicting delayed graft function in deceased donor kidney transplantation based on pre-transplant biopsies.

BACKGROUND: Delayed graft function (DGF) is an important complication after kidney transplantation surgery. The present study aimed to develop and validate a nomogram for preoperative prediction of DGF on the basis of clinical and histological risk factors. METHODS: The prediction model was constructed in a development cohort comprising 492 kidney transplant recipients from May 2018 to December 2019. Data regarding donor and recipient characteristics, pre-transplantation biopsy results, and machine perfusion parameters were collected, and univariate analysis was performed. The least absolute shrinkage and selection operator regression model was used for variable selection. The prediction model was developed by multivariate logistic regression analysis and presented as a nomogram. An external validation cohort comprising 105 transplantation cases from January 2020 to April 2020 was included in the analysis. RESULTS: 266 donors were included in the development cohort, 458 kidneys (93.1%) were preserved by hypothermic machine perfusion (HMP), 96 (19.51%) of 492 recipients developed DGF. Twenty-eight variables measured before transplantation surgery were included in the LASSO regression model. The nomogram consisted of 12 variables from donor characteristics, pre-transplantation biopsy results and machine perfusion parameters. Internal and external validation showed good discrimination and calibration of the nomogram, with Area Under Curve (AUC) 0.83 (95%CI, 0.78-0.88) and 0.87 (95%CI, 0.80-0.94). Decision curve analysis demonstrated that the nomogram was clinically useful. CONCLUSION: A DGF predicting nomogram was developed that incorporated donor characteristics, pre-transplantation biopsy results, and machine perfusion parameters. This nomogram can be conveniently used for preoperative individualized prediction of DGF in kidney transplant recipients.

Knockdown of ANGPTL2 Protects Renal Tubular Epithelial Cells Against Hypoxia/Reoxygenation-Induced Injury via Suppressing TLR4/NF-κB Signaling Pathway and Activating Nrf2/HO-1 Signaling Pathway.

Renal ischemia/reperfusion (I/R) injury is a particular threat faced by clinicians in kidney transplantation. Previous studies have confirmed the importance of oxidative stress and inflammation in the pathogenesis of I/R injury. Angiopoietin-like protein 2 (ANGPTL2) belongs to the angiopoietin-like family and has been found to be involved in the regulation of kidney function as well as oxidative and inflammatory response. In the present study, we aimed to evaluate the role of ANGPTL2 in renal I/R injury in vitro. The human proximal tubular epithelial cell line (HK-2 cells) was subjected to hypoxia/ reoxygenation (H/R) to mimic I/R injury in vitro. We found that the expression level of ANGPTL2 was markedly increased in H/R-induced HK-2 cells. Knockdown of ANGPTL2 improved the decreased cell viability of HK-2 cells in response to H/R stimulation. Knockdown of ANGPTL2 significantly inhibited the H/R-caused increase in levels of reactive oxygen species, malondialdehyde, and proinflammatory cytokines, including interleukin (IL)-6, IL-1ß, and tumor necrosis factor-alpha, as well as a decrease in superoxide dismutase activity in the HK-2 cells. Besides, the increased bax expression and caspase-3 activity and decreased bcl-2 expression in H/R-induced HK-2 cells were also attenuated by knockdown of ANGPTL2. Moreover, ANGPTL2 overexpression showed the opposite effects. Further mechanism investigations proved that the activation of Nrf2/HO-1 signaling pathway and the inhibition of toll-like receptor 4/nuclear factor kappa-light-chain-enhancer of activated B cells signaling pathway were both implicated in the renal-protective effects of ANGPTL2 knockdown on H/R-induced HK-2 cells. Collectively, these findings suggested that ANGPTL2 might be a new possible target for the treatment and prevention of renal I/R injury.

Comprehensive assessment of deceased donor kidneys with clinical characteristics, pre-implant biopsy histopathology and hypothermic mechanical perfusion parameters is highly predictive of delayed graft function.

Background: Due to the current high demand for transplant tissue, an increasing proportion of kidney donors are considered extended criteria donors, which results in a higher incidence of delayed graft function (DGF) in organ recipients. Therefore, it is important to fully investigate the risk factors of DGF, and establish a prediction system to assess donor kidney quality before transplantation.Methods: A total of 333 donation after cardiac death kidney transplant recipients were included in this retrospective study. Both univariate and multivariate analyses were used to analyze the risk factors of DGF occurrence. Receiver operating characteristic (ROC) curves were used to analyze the predictive value of variables on DGF posttransplant.Results: The donor clinical scores, kidney histopathologic Remuzzi scores and hypothermic mechanical perfusion (HMP) parameters (flow and resistance index) were all correlated. 46 recipients developed DGF postoperatively, with an incidence of 13.8% (46/333). Multivariate logistic regression analysis of the kidney transplants revealed that the independent risk factors of DGF occurrence post-transplantation included donor score (OR = 1.12, 95% CI 1.06-1.19, p < 0.001), Remuzzi score (OR = 1.21, 95% CI 1.02-1.43, p = 0.029) and acute tubular injury (ATI) score (OR = 4.72, 95% CI 2.32-9.60, p < 0.001). Prediction of DGF with ROC curve showed that the area under the curve was increased to 0.89 when all variables (donor score, Remuzzi score, ATI score and HMP resistance index) were considered together.Conclusions: Combination of donor clinical information, kidney pre-implant histopathology and HMP parameters provide a more accurate prediction of DGF occurrence post-transplantation than any of the measures alone.

C1q/TNF-related protein 6 (CTRP6) attenuates renal ischaemia-reperfusion injury through the activation of PI3K/Akt signalling pathway.

C1q/TNF-related protein 6 (CTRP6) is a member of the CTRP family that has been reported to exhibit a nephroprotective effect. However, the role of CTRP6 in renal ischaemia/reperfusion (I/R) injury (IRI) remains unclear. In the present study, we aimed to explore the protective effect of CTRP6 in renal IRI and the potential mechanism. We found that CTRP6 expression was markedly decreased in the kidneys of mice subjected to I/R and HK-2 cells in response to hypoxia/reoxygenation (H/R) stimulation. Recombinant CTRP6 protein protected against renal I/R injury by the reduction of blood urea nitrogen (BUN) and creatinine levels. The increased production of ROS and malondialdehyde (MDA), as well the decreased activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD) caused by H/R induction were mitigated by CTRP6 in HK-2 cells. The caspase-3 activity and apoptotic rate were both decreased in CTRP6-overexpressing HK-2 cells. In addition, we also found that knockdown of CTRP6 aggravated H/R-caused oxidative stress and cell apoptosis in HK-2 cells. Moreover, CTRP6 overexpression enhanced the H/R-stimulated activation of PI3K/Akt pathway in HK-2 cells. Inhibition of PI3K reversed the nephroprotective effects of CTRP6 in HK-2 cells. Taken together, CTRP6 exerted protective effects against H/R-caused oxidative injury in HK-2 cells via activating the PI3K/Akt pathway.

Prediction of kidney transplant outcome based on different DGF definitions in Chinese deceased donation.

BACKGROUND: Delayed graft function (DGF) is an important complication of kidney transplantation and can be diagnosed according to different definitions. DGF has been suggested to be associated with the long-term outcome of kidney transplantation surgery. However, the best DGF definition for predicting renal transplant outcomes in Chinese donations after cardiac death (DCDs) remains to be determined. METHOD: A total of 372 DCD kidney transplant recipients from June 2013 to July 2017 in the First Affiliated Hospital of Xi'an Jiaotong University were included in this retrospective study to compare 6 different DGF definitions. The relationships of the DGF definitions with transplant outcome were analyzed, including graft loss (GL) and death-censored graft loss (death-censored GL). Renal function indicators, including one-year estimated glomerular filtration rate (eGFR) and three-year eGFR, and were compared between different DGF groups. RESULTS: The incidence of DGF varied from 4.19 to 35.22% according to the different DGF diagnoses. All DGF definitions were significantly associated with three-year GL as well as death-censored GL. DGF based on requirement of hemodialysis within the first week had the best predictive value for GL (AUC 0.77), and DGF based on sCr variation during the first 3 days post-transplant had the best predictive value for three-year death-censored GL (AUC 0.79). Combination of the 48-h sCr reduction ratio and classical DGF can improve the AUC for GL (AUC 0.85) as well as the predictive accuracy for death-censored GL (83.3%). CONCLUSION: DGF was an independent risk factor for poor transplant outcome. The combination of need for hemodialysis within the first week and the 48-h serum creatinine reduction rate has a better predictive value for patient and poor graft outcome.

FOXP1 inhibits high glucose-induced ECM accumulation and oxidative stress in mesangial cells.

Diabetic nephropathy (DN) is a common complication of diabetes that remains the major cause of end-stage renal disease (ESRD). Forkhead box P1 (FOXP1) is a member of FOX family involved in the progression of diabetes. However, the pathogenic role of FOXP1 in DN remains unclear. This study was aimed to explore the effects of FOXP1 on glomerular mesangial cells (MCs) in response to high glucose (HG) stimulation. We found that HG stimulation markedly inhibited the FOXP1 expression in MCs in dose-and time-dependent manner. CCK-8 assay proved that FOXP1 overexpression attenuated HG-induced cell proliferation in MCs. FOXP1 exhibited anti-oxidative activity in HG-induced MCs, as proved by the decreased production of ROS and expressions of ROS producing enzymes, NADPH oxidase (NOX) 2 and NOX4. Besides, FOXP1 suppressed the expression and secretion of extracellular matrix (ECM) proteins including collagen IV (Col IV) and fibronectin (FN). Furthermore, FOXP1 overexpression significantly prevented HG-induced activation of Akt/mTOR signaling in MCs, and Akt activator blocked FOXP1-mediated cell proliferation, ROS production and ECM accumulation in MCs. Collectively, FOXP1 prevented HG-induced proliferation, oxidative stress, and ECM accumulation in MCs via inhibiting the activation of Akt/mTOR signaling pathway. The findings suggested that FOXP1 might be a therapeutic target for the treatment of DN.

Role of prostaglandin E2 receptor 4 in the modulation of apoptosis and mitophagy during ischemia/reperfusion injury in the kidney.

The mechanisms by which prostaglandin E2 receptor 4 (EP4) protects against renal ischemia­reperfusion (I/R) injury (IRI) remain to be fully elucidated. In the present study, the protective effects of EP4 signaling on renal mitochondria and against renal IRI, as well as the underlying mechanisms, were investigated. A rat model of renal IRI was established. The right kidney was separated without damaging the artery clip, and the renal blood perfusion was then restored after 60 min. One group of animals was treated with EP4 agonists prior to I/R. The mitochondrial mass, the copy number of mitochondrial (mt)DNA, adenosine triphosphate (ATP) production and mitochondrial autophagy were analyzed. It was identified that renal IRI reduced the mitochondrial mass, decreased the mtDNA copy number and inhibited ATP production. The loss of renal mitochondria was attributed to the excessive mitochondrial autophagy induced by renal IRI. Pre­treatment with EP4 agonist inhibited excessive mitochondrial autophagy, the loss of mitochondria and maintained and the energy imbalance within the cells. It was indicated that renal IRI causes excessive mitochondrial autophagy, which is one of the important causes of renal dysfunction.

Probiotics ameliorate renal ischemia-reperfusion injury by modulating the phenotype of macrophages through the IL-10/GSK-3ß/PTEN signaling pathway.

After renal ischemic reperfusion injury, a series of pathological changes, such as impaired intestinal barrier function, intestinal flora, and endotoxin translocation, are caused by intestinal ischemia and hypoxia, which then trigger systemic inflammatory responses and affect the condition and prognosis of the patients. In this study, a rat model of ischemia-reperfusion injury was established by examining changes in renal function, intestinal barrier function, inflammatory index, oxidative stress, and macrophage phenotypes to evaluate the effect of probiotic VSL#3 on renal ischemia-reperfusion injury. The results showed that, after VSL#3 intervention, the levels of BUN, Scr, Cys C, PRO, and NGAL were all significantly decreased compared with the I/R group, while the value of Ccr showed a significant increase. In addition, the concentrations of MPO, IL-1ß, TNF-α, IL-6, ED-1, and PCNA were all significantly lower than those in the I/R group, while the levels of endotoxin, DOA, and D-lactic acid were significantly decreased. Furthermore, the proteins associated with intestinal barrier functions, such as ZO-1, Occludin, and Claudin-1, were significantly upregulated compared with the I/R group. Overall, the VSL#3 intervention group was able to maintain the required number of beneficial intestinal flora and to inhibit the proliferation of harmful bacteria. At the same time, the VSL#3 intervention could also prevent the decrease in the levels of CAT, GSH-PX, H2O2, and T-SOD, while downregulating the expression of Keap1 and Nrf2. After the intervention with the VSL#3, the expression levels of CD68 and CD86 proteins were significantly decreased, while the expression levels of CD163 and CD206 proteins were significantly higher. Further experiments confirmed that the expression of iNOS protein was significantly decreased after the VSL#3 intervention, and the expression of Arg-1 and Ym1 proteins was significantly increased. The VSL#3 was able to induce high expressions of p-GSK-3ß and p-PTEN proteins, while the use of IL-10 antibody impaired the effect of the VSL#3. In summary, this research confirms that probiotics can alleviate renal dysfunction caused by ischemia and reperfusion by protecting the intestinal barrier function and maintaining the functions of intestinal flora. The pathway screening test of this study suggests that IL-10/GSK3ß/PTEN may play an important role in the process of the prototypic VSL#3 inducing M2 transformation of macrophages.

LncRNA CRNDE is a biomarker for clinical progression and poor prognosis in clear cell renal cell carcinoma.

Colorectal neoplasia differentially expressed (CRNDE) served as an oncogenic long noncoding RNA (lncRNA) to be involved in the initialization and development of human cancers. However, the clinical significance and biological function of CRNDE in clear cell renal cell carcinoma (ccRCC) was not fully understood. In our study, we found CRNDE levels were increased in ccRCC tissue specimens and cell lines, and corrected with advanced clinical stage, large tumor size, lymph node metastasis, distant metastasis, and poor pathological grade in patients with ccRCC. Furthermore, levels of CRNDE were negatively correlated with overall survival of patients with ccRCC, and high-expression of CRNDE was an independent poor prognostic factor for patients with ccRCC. Moreover, loss-of-function and gain-of-function approaches showed CRNDE-enhanced ccRCC cell migration and invasion through modulating EMT-associated genes. In conclusion, CRNDE acts as an oncogenic lncRNA in ccRCC.

Early Immunosuppressive Exposure of Enteric-Coated-Mycophenolate Sodium Plus Tacrolimus Associated with Acute Rejection in Expanded Criteria Donor Kidney Transplantation.

BACKGROUND: Immunosuppressive agents are still inefficient in preventing biopsy-proven acute rejection (BPAR) after expanded criteria donor (ECD) kidney transplantation. The aim of this study was to investigate the relationships between early immunosuppressive exposure and the development of BPAR. METHODS: We performed a retrospective study of 58 recipients of ECD kidney transplantation treated with enteric-coated-mycophenolate sodium, tacrolimus (Tac), and prednisone. The levels of mycophenolic acid-area under the curve (MPA-AUC)0-12h and Tac C0were measured at the 1st week and the 1st month posttransplant, respectively. The correlation was assessed by multivariate logistic regression. RESULTS: The occurrence rates of BPAR and antibody-mediated rejection were 24.1% and 10.3%, respectively. A low level of MPA-AUC0-12h at the 1st week posttransplant was found in BPAR recipients (38.42 ± 8.37 vs. 50.64 ± 13.22, P < 0.01). In addition, the incidence of BPAR was significantly high (P < 0.05) when the MPA-AUC0-12hlevel was <30 mg·h-1·L-1 at the 1st week (15.0% vs. 44.4%) or the Tac C0was <4 ng/ml at the 1st month posttransplant (33.3% vs. 21.6%). Multivariable logistic regression analysis showed that the MPA-AUC0-12h at the 1st week (OR: 0.842, 95% CI: 0.784-0.903) and the Tac C0at the 1st month (OR: 0.904, 95% CI: 0.822-0.986) had significant inverse correlation with BPAR (P < 0.05). CONCLUSIONS: Low-level exposure of MPA and Tac C0in the early weeks posttransplant reflects an increased acute rejection risk, which suggested that MPA-AUC0-12h <30 mg·h-1·L-1 and Tac C0 <4 ng/ml should be avoided in the first few weeks after transplantation.

Current attitudes toward organ donation after cardiac death in northwest China.

BACKGROUND: People's attitude toward organ donation after cardiac death (DCD) has not come to an agreement in different countries and regions. Influenced by the local culture in China for thousands of years, the general public has different ideas about this issue. The purpose of this study was to investigate the current attitudes trend and characteristics of transplantation with organs donated after cardiac death in northwest China. METHODS: This largest single-center cohort study was performed by an interview or by telephone using a questionnaire. The family members of potential DCD donors were recruited from the First Affiliated Hospital, medical college of Xi'an Jiaotong University located in a metropolitan area of northwest China. The 12-item attitude questionnaire was specifically developed from the literature review with coordinator, physician, and donor's family feedback. The participants were asked to rate the queries on a 5-point Likert intensity scale. RESULTS: The 174 participants included 56 (32.2%) women and 118 (67.8%) men. Most people were aged between 41 and 50 years (n = 63, 36.2%), 31 and 40 years (n = 59, 33.9%), and less than 30 years (n = 36, 20.7%). The top five attitudes of participants were the best person to suggest organ donation to a family was ranked as the DCD coordinator of Red Cross Organization (RCO, n = 160, 92%), donor is a hero (n = 143, 82.2%), honor to be a donor's family member (n = 136, 78.2%), improved relationship with colleagues (n = 124, 71.3%), and with recipient after donation (n = 123, 70.7%). The best person to suggest organ donation to a family was ranked as the coordinator of RCO (n = 160, 92%), doctor unrelated to transplantation (n = 104, 59.8%), social worker (n = 36, 20.7%), and doctor related to transplantation (n = 25, 14.4%). The top two reasons for non-consent to donation were that the family insisted on intact body after patient death and did not want to have surgery again (n = 51, 41.5%), and feared that they would be misunderstood by neighbors, relatives, and friends about donation (n = 28, 22.8%). CONCLUSIONS: This study revealed initial attitudes toward DCD in China. Some data afford insight into the decision-making procedure. The concerns of potential DCD donors and their families may help professionals provide better interventions in the future.

Assessment of different biomarkers provides valuable diagnostic standards in the evaluation of the risk of acute rejection.

Acute rejection (AR) is a strong risk factor for chronic rejection in renal transplant recipients. Accurate and timely diagnosis of AR episodes is very important for disease control and prognosis. Therefore, objectively evaluated the immune status of patients is essential in the field of post-transplantation treatment. This longitudinal study investigated the usefulness of five biomarkers, human leukocyte antigen (HLA)-G5 and sCD30 level in sera, intracellular adenosine triphosphate (iATP) release level of CD4(+) T cells, and granzyme B/perforin expression in peripheral blood mononuclear cells (PBMCs) and biopsies, to detect AR and the resolution of biomarkers in a total of 84 cases of renal transplantation. The data demonstrated that recipients with clinical or biopsy proven rejection significantly increased iATP release level of CD4(+) T cells, and elevated sCD30 but lowered HLA-G5 level in sera compared with individuals with stable graft function. Expression levels of granzyme B and perforin were also elevated in PBMCs and graft biopsies of AR patients. Taken together, we identified that upregulation of sCD30, iATP, granzyme B, perforin, and downregulation of HLA-G5 could provide valuable diagnostic standards to identify those recipients in the risk of AR. And iATP may be a better biomarker than others for predicting the graft rejection episode.

Hypoxic preconditioning advances CXCR4 and CXCR7 expression by activating HIF-1α in MSCs.

Recent evidence indicated that sublethal hypoxic preconditioning (HP) of bone marrow-derived mesenchymal stem cells (MSCs) before transplantation could ameliorate their capacity to survive and engraft in the target tissue through yet undefined mechanisms. In this study, we demonstrated that HP (3% oxygen) induced the high expression of both chemokine stromal-derived factor-1 (SDF-1) receptors, CXCR4 and CXCR7, in MSCs. HP also improved in vitro migration, adhesion and survival of MSCs. Although SDF-1-induced migration of HP-MSCs was only abolished by an anti-CXCR4 antibody, both CXCR4 and CXCR7 were responsible for elevated adhesion of HP-MSCs. Moreover, CXCR7 but not CXCR4 was essential for the resistance to oxidative stress of HP-MSC. In addition, HP also evoked an increase in expression of hypoxia-inducible factor-1 (HIF-1α) and phosphorylation of Akt. The chemical inducers of HIF-1α, desferrioxamine (DFX) and cobalt chloride (CoCl2), induced upregulation of CXCR4 and CXCR7 expression in MSCs under normoxic conditions. Contrarily, blockade of HIF-1α by siRNA and inhibition of Akt by either wortmannin or LY294002 abrogated upregulation of HP-induced CXCR4 and CXCR7 in MSCs. Collectively, these findings provide evidence for a crucial role of PI3K/Akt-HIF-1α-CXCR4/CXCR7 pathway on enhanced migration, adhesion and survival of HP-MSCs in vitro.

[Living-related donor kidney transplantation in 158 patients].

OBJECTIVE: To introduce clinical experience for living-related donor kidney transplantation (LDKT) by reviewing LDKT clinical data. METHODS: A total of 158 patients underwent LDKT. Expect for 7 patients donated by their spouses, the others had blood relationship donors. Donor-recipient HLA matching showed 2 patients had 5-loci mismatch, 5 with 4-loci mismatch, 88 with 3-loci mismatch, 50 with 2-loci mismatch, 12 with 1-loci mismatch, the other 1 with 0-loci mismatch. All of the 158 donors underwent open nephrectomy, 35 of whom donated the right kidneys and the other 123 donated the left kidneys. Triple immunosuppressive regimen consisted of calcineurin inhibitors or FK506, MMF or AZa, and steroid. RESULTS: All donors were healthy after the operation. All donors were followed up for 6 to 12 months and blood exams showed that inosine levels were normal. The longest kidney transplant functional survival time was 10 years to up June 2008. The one year patient/graft survival rate was 95.5%. Delayed graft function (DGF) occurred in 5 patients, 4 of whom recovered in 2-5 weeks. Five patients died, 4 of whom died of post-operational pulmonary infection within 3-5 months, with no transplantational complications. The other one died of pulmonary bleeding during dialysis while treating for DGF. One patient received a second deceased kidney transplant because of hyperacute rejection during the surgery. Five developed acute rejection 1 month after the operation (incidence rate 3.16%), 4 of whom were cured by administration of methylprednisolone, and the other one returned to dialysis because of renal toxicity of cyclosporine. Three patients had positive chronic rejection, 2 of whom lost graft function in 1.5-3.5 years. Eight patients developed pulmonary infection and 4 of them were cured. CONCLUSION: Sufficient LDKT pre-operational assessment, satisfactory tissue matching and reduced ischemia time may result in lower incidence of DGF, acute rejection and higher patient/graft survival rate. In LDKT, importance should also be attached to the prevention of DGF and graft rejection. Rational dosage of immunosuppressants is advocated to prevent secondary infective complications. Donor specifications and all around evaluation of the living-related donors should also be emphasized to minimize the harm to the donors. Long term follow-up is also essential to ensure donors' post-operational healthy life.

[Follow-up evaluation of a new ureteral anastomosis technique in renal transplantation].

OBJECTIVE: To compare the therapeutic effect of new "One-Stitch" ureterovesical anastomosis to that of the classic Lich-Gregoir method. METHODS: From January 2002 to December 2004, 445 patients suffering from uremia due to chronic nephritis were treated with renal transplantation. Among them, 121 patients were operated with our new modified ureteroneocystostomy technique (the new One-Stitch group), and 324 patients were operated with Lich-Gregoir technique (the Lich-Gregoir group). In the new One-Stitch group, there were 79 males and 42 females, aged 20-62 years (35.7 years on average). The course of the disease was 2-11 years (2.7 years on average). In the Lich-Gregoir group, there were 211 males and 113 females, aged 19-65 years (33.9 years on average). The disease course was 1-14 years (2.3 years on average). There was no significant difference between the two groups in age, proportion of genders, primary diseases and course of the disease (P > 0.05). The operative time, the ureteral complications and non-ureteral complications were compared between the two groups after the renal transplantation. RESULTS: The operative time for the new One-Stitch and Lich-Gregoir techniques was (8.7 +/- 1.1) minutes and (22.4 +/- 5.1) minutes, indicating the difference was significant (P < 0.05). All recipients were followed up for 3-5 years. In the new One-Stitch group, there were 5 patients with leakage of urine, 15 with gross hematuria, 4 with ureteral obstruction and 28 with urinary system infection. Symptomatic vesicoureteral reflux and stone formation were not observed in this group. In the Lich-Gregoir group, there were 17 patients with leakage of urine, 12 with gross hematuria, 13 with ureteral obstruction, 86 with urinary system infection, 6 with symptomatic vesicoureteral reflux and 2 with stones. In the new One-Stitch group, the incidence rate of complications of gross hematuria was 12.4%, which was significantly different from 3.7% in the Lich-Gregoir group (P < 0.05). The incidence rates of ureteral complications in the Lich-Gregoir and the new One-Stitch groups were 19.8% and 15.4%, respectively. The difference was not significant (P > 0.05). There was no significant difference between the two groups in incidence rate of urinary system infection, delayed recovery of kidney function after kidney transplantation and rejection reaction (P > 0.05). CONCLUSION: The new One-Stitch group has no significant difference in ureteral complications compared with the Lich-Gregoir group, and has become a preferential ureterovesical reimplantation technique because of its simple and has convenient operation.

[Recombinant adenovirus-mediated human cytosolic glutathione peroxidase gene transfection protects vascular endothelial cells from oxidative damage].

OBJECTIVE: To study the protective effect of recombinant adenovirus-mediated human cytosolic glutathione peroxidase (hCGPx) gene transfection on vascular endothelial cells ECV304 from oxidative damage. METHODS: pGEM-T Easy Vector containing hCGPx cDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were used to construct the shuttle plasmid pACCMV-hCGPx for cotransfection of 293 cells with pJM17, thereby to obtain the recombinant adenovirus AdCMV-hCGPx. Cultured ECV304 cells were transfected with AdCMV-hCGPx for 24, 48 and 72 h, respectively, with the cells transfected with the empty vector serving as control, and hCGPx gene expression was then examined in the transfected cells. The transfected cell viability and apoptotic cell ratio were evaluated after treatment of the cells with H(2)O(2). RESULTS: The expression ratio of hCGPx gene was significantly higher in the AdCMV-hCGPx-transfected cells than in those with empty vector transfection (P<0.01). The hCGPx gene-transfected cells showed significantly higher viability and significantly lower apoptotic ratio than the control cells following challenge with H(2)O(2)-induced oxidative damage. CONCLUSION: hCGPx gene transfer mediated by recombinant adenovirus protects the vascular endothelial cells from oxidative damage in vitro, possibly due to the antioxidative and apoptosis-inhibiting effect of hCGPx.

[Protection of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by recombinant adenovirus containing hCGPx gene].

AIM: To study the protective effect of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by transfection of recombinant adenovirus-mediated human cytosolic glutathione peroxidase(hCGPx) gene. METHODS: Recombinant pGEM-T vector containing hCGPx cDNA and pACCMV-pLpA adenovirus shuttle plasmid was constructed. Then the shuttle plasmid pACCMV-hCGPx and pJM17 were co-transfected into 293 cells and recombinant adenovirus AdCMV-hCGPx was obtained. Cultured HK-2 cells were transfected with AdCMV-hCGPx or vacant recombinant adenovirus (control). The expression ratio of transfected hCGPx gene were studied. Cell viability, the percentage of apoptosis and death were evaluated after hypoxia-reoxygenation damage. RESULTS: The expression ratio of hCGPx gene was higher in the AdCMV-hCGPx transfected cells than that in the control group (P<0.01). After hypoxia-reoxygenation damage, the viability of hCGPx gene transfected cells was significantly higher than that of control and the percentage of apoptosis and death of hCGPx transfected cells was significantly lower than that of control. CONCLUSION: The transfection of hCGPx mediated by recombinant adenovirus could protect renal tubular epithelial cells from hypoxia-reoxygenation damage in vitro.