Risk factors for urinary tract infection in kidney transplantation from brain death donor and its role in graft function / 中南大学学报(医学版)

OBJECTIVES@#Urinary tract infection (UTI) is the most common infection complication after kidney transplantation, and the reports of the incidence vary greatly among different centers. This study aims to explore the risk factors for UTI after kidney transplantation with the donation from brain death (DBD) and the impact on graft function, thus to provide theoretical basis for comprehensive prevention and treatment of UTI after kidney transplantation.@*METHODS@#The clinical and laboratory data of DBD kidney transplantation from January 2017 to December 2018 in Xiangya Hospital, Central South University were collected and retrospectively analyzed. Patients were assigned into an UTI group and a non-UTI group. The base line characteristics, post-transplant complications, and graft function were compared between the 2 groups. Multivariate logistic regression was used to analyze the risk factors for UTI.@*RESULTS@#A total of 212 DBD kidney transplant recipients were enrolled in this study. UTI occurred in 44 (20.75%) patients after transplantation. The female, the time of indwelling catheter, and postoperative urinary fistula were independent risk factors for UTI after DBD kidney transplantation. A total of 19 strains of gram-positive bacteria, 12 strains of gram-negative bacteria , and 10 strains of fungi were isolated from the urine of 44 UTI patients. The UTI after kidney transplantation significantly increased time of hospital stay (@*CONCLUSIONS@#UTI after DBD kidney transplantation transplantation affects the renal function at 3 months and increases the patient's economic burden.

Transient stopping immunosuppressive agents during the post-transplant pulmonary infection does not affect the long-term outcome of renal transplantation / 中南大学学报(医学版)

OBJECTIVE@#To determine the effect of transient withdrawal of immunosuppressive agents during the treatment of pulmonary infection on long-term survival of patients and graft s.@*METHODS@#A total of 104 patients with post-transplant pulmonary infection were enrolled in this study. These patients received renal transplantation in Center for Organ Transplantation, Xiangya Hospital, Central South University, during December 2005 and August 2014. Among them, 50 patients stopped immunosuppressive agents during the treatment of infection. These patients served as stopping drug (SD) group, whereas the remaining patients who served as a control group did not stop immunosuppressive drugs. The five-year cumulative patient survival, graft survival, and laboratory results were compared between the 2 groups.@*RESULTS@#The five-year cumulative patient survival rates in the SD group were significantly lower than those in the control group [(69.8 ± 7.0)% vs (94.2 ± 3.2)%, P=0.001]. There was no significant difference in the allograft survival rates between the 2 groups [(81.7 ± 6.6)% vs (90.9 ± 4.3)%, P=0.113]. In patients who survived from pulmonary infection, there was no significant difference in long-term survival rates between the 2 groups (P=0.979).@*CONCLUSION@#Pulmonary infection impacts allograft survival after patients underwent renal transplantation. Transient stopping immunosuppressive agents during the treatment of infection is a safe and necessary treatment strategy for patients with serious post-transplant pulmonary infection.

Fibronectin splice variant connecting segment-1 peptides protect sinusoidal endothelial cells and alleviate ischemia/reperfusion injury in rats / 中华器官移植杂志

Objective To examine the effect of fibronectin connecting segment-1 (CS1) peptidefacilitated blockade of inflammatory cells-fibronectin adhesion on a rat liver transplantation model of prolonged ex vivo cold ischemia.Methods A model of liver transplantation in Wistar→Wistar rat was established.The donors of the CS1 treatment group received CS1 peptides through the tail vein for 3 days before operation.Another two doses of CS1 peptides were administered into the liver intraportally during procurement and before transplantion.Recipients received an additional 3-day course of CS1 peptides after transplantation.Rats in control group received scrambled peptides.Rats were sacrificed at 6,24 and 72 h after transplantion,and plasma transaminase activity and hepatic pathological changes were studied.The inflammatory cells and liver sinusoidal endothelial cells were visualized histochemically.Real-time PCR was used to detect tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and vascular endothelial growth factor (VEGF) mRNA expression in the liver.Results The plasma transaminase activity and hepatic necrosis areas in CS1 treatment group were significantly lower than in control group (P＜0.05).CS1 peptides treatment significantly decreased the number of Kupffer cells after transplantation and greatly inhibited the recruitment of neutrophils to the graft liver as compared with control group (P＜0.05).After prolonged cold ischemia,only a few hepatic endothelial cells exhibited positive staining of hepatic sinusoidal endothelial cell biomarker SE-1.Lots of hepatic sinusoidal endothelial cells positive for SE-1 staining could be detected in CS1 group at 72 h after transplantation,while much less SE-1 positive cells presented in the control goup.Prolonged cold ischemia caused a significant increase of TNF-α,IL-1β and VEGF mRNA expression in the graft liver of control group after transplantation.The expression of TNF-α mRNA at 6 and 24 h and VEGF mRNA expression at 24 h were significantly lower in CS1 group than in control group (P＜0.05).Conclusion Peptide-mediated blockade of inflammatory cells-fibronectin interaction decreased the mRNA expression of inflammatory cytokines,prevented hepatic sinusoidal endothelial cells from injury and subsequently protected against severe ischemia/reperfusion injury of the graft liver after transplantation.

Effect of gene GSTP1 silencing via shRNA transfection on androgen independent prostate cancer cell line Du145 / 中南大学学报(医学版)

OBJECTIVE@#To design short hairpin RNA (shRNA) interference sequence to silence glutathione S-transferase P1 (GSTP1) gene of androgen independent prostate cancer cell line DU145, and to explore its effect on proliferation and sensitivity to chemotherapeutics.@*METHODS@#The target sequence was picked up to form the shRNA, and the 3 shRNA expression vectors were shRNA255, shRNA554 and shRNA593. The DNA template was cloned to plasmid pGPU6/GFP/Neo. The shRNA was identified by enzyme digesting and gene sequencing. The screening experiment was done to pick up the shRNA expression vector with the highest transfection ratio and best gene silencing results. DU145 cells were divided into a blank plasmid group and a shRNA transfected group. According to the chemotherapeutics the DU145 cells were divided into a fluorouracil (FU) group and a paclitaxel (PA) group, and the 2 groups were subdivided into 4 subsets according to the chemotherapeutic concentrations (FU: 30, 60, 120, and 240 μg/mL; PA: 0.2, 2, 10, and 20 μg/mL), meanwhile a blank control group was included respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the proliferation after the transfection. MTT and terminal de-oxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were used to detect the inhibition effect of different concentrations of 5-FU or PA on the proliferation and induction of apoptosis of DU145.@*RESULTS@#The transfection ratio of the 3 shRNA expression vectors (shRNA255, shRNA554, and shRNA593) was (63.30±1.04)%, (76.20±0.68)%, and (72.70±0.33)%, and the transfection ratio of shRNA554 was the highest. there was significant difference among the above 3 shRNA expression vectors (P<0.01). After the transfection, the mRNA was 128.31±2.50, 43.24±4.30 and 85.62±6.30, the GSTP1 protein was 163.92±12.40, 65.38±9.30 and 114.25±16.70. After the transfection of shRNA554, the mRNA and protein of GSTP1 were the lowest level. there was significant difference among the above 3 shRNA expression vector (P<0.01). MTT analysis showed that before the transfection, the survival ratio of cells under different concentrations of FU (30, 60, 120, and 240 μg/mL) was (95.60±2.11)%, (90.20±0.86)%, (83.10±3.12)% and (74.60±1.32)%; however after the transfection, the survival ratio of cells was (91.30±1.43)%, (84.60±2.13)%, (73.20±1.52)%, and (65.5±0.942)%. TUNEL assay showed that before the transfection, the apoptosis ratio of cells under different concentrations of FU (30, 60, 120, and 240 μg/mL) was (5.50±0.88)%, (10.20±1.64)%, (15.20±2.39)%, and (25.10±2.59)%; however after the transfection, the apoptosis ratio of cells was (10.8±0.62)%, (15.7±1.32)%, (20.4±1.89)%, and (34.9±2.54)%. After the transfection, the cell survival ratio decreased under the same concentration of FU, and the apoptosis ratio increased, with statistical significance (both P<0.01). MTT analysis showed that before the transfection, the survival ratio of cells under different concentrations of PA (0.2, 2, 10, and 20 μg/mL) was (98.50±2.34)%, (95.20±1.32)%, (89.40±0.68)%, and (82.70±1.73)%; after the transfection the survival ratio of cells was (94.20±0.78)%, (86.50±2.13)%, (78.70±1.34)%, and (70.10±0.76)%. TUNEL assay showed that before the transfection, the apoptosis ratio of cells under different concentrations of PA (0.2, 2, 10, and 20 μg/mL) were (2.40±1.07)%, (5.20±1.33)%, (10.50±2.41)%, (20.70±1.92)%; after the transfection the apoptosis ratio of cells was (5.46±2.13)%, (13.80±1.24)%, (21.20±2.39)%, and (29.20±2.21)%. After the transfection, the cell survival ratio decreased under the same PA concentration, and the apoptosis ratio increased, with statistical significance (both P<0.01).@*CONCLUSION@#gene GSTP1 silence via shRNA transfection to androgen independent prostate cancer cell line DU145 can inhibit its proliferation in time dependent manner, and induce apoptosis and raise its sensitivity to chemotherapeutics.

Expression of Foxp3 mRNA in peripheral blood monocytes in patients after kidney transplantation / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationship between the level of Foxp3 mRNA in the peripheral blood monocytes (PBMCs) with acute rejection or chronic allograft nephropathy after kidney allotransplantation.@*METHODS@#Real-time quantitative polymerase chain reaction was used to examine Foxp3 mRNA expression in the PBMCs from 16 patients with acute rejection, 8 patients with chronic allograft nephropathy, 8 patients at stable stage after kidney transplantation, 8 patients of end-stage renal disease, and 8 normal controls.@*RESULTS@#The level of Foxp3 mRNA in the PBMCs was significantly lower in patients with acute rejection than that in patients with chronic allograft nephropathy (P0.05).@*CONCLUSION@#The level of Foxp3 mRNA expressed in PBMCs can reflect the status of renal allograft, and may be a noninvasive biomarker for diagnosing acute rejection and chronic allograft nephropathy.