Objective    To retrospectively study the risk factors of aortic arch calcificationand its influence on the survival prognosis of maintenance peritoneal dialysis patients.    Methods    One hundred seventy-seven cases of maintenance peritoneal dialysis patients were enrolled, including 66 cases of aortic arch calcification cases. Their general dialysis data were collected for the evaluation of dialysis adequacy and residual renal function, and their chest X-rays were recorded to assess the degree of aortic arch calcification. The two variables Logistics regression was used to analyze independent risk factors of aortic arch calcification; Kaplan-Meier analysis was used to analyze the influence on prognosis of dialysis patients; and multivariate COX regression was employed to analyze independent risk factors of death in dialysis patients.    Results    Among the 177 selected cases of peritoneal dialysis patients, 66 cases (37.29%) presented with aortic arch calcification. Elevated serum phosphorus was an independent risk factor of aortic arch calcification (OR=54.69，95%CI: 10.01-298.65, P＜0.01). The probability of survival in patients with mild and moderate (severe) calcification of aortic arch was less than those without calcification. Moderate (severe) calcification of aortic arch was the independent risk factor of all-cause mortality and cardiovascular disease mortality, whose hazard ratios in patients with calcification were 3.779 times and 5.636 times of those in patients without calcification respectively.    Conclusions    Hyperphosphatemia is an independent risk factor promoting the development of calcification of aortic arch. The probability of survival in patients with mild and moderate (severe) calcification of aortic arch is less than those without calcification; moderate (severe) calcification of aortic arch is the independent risk factor of all-cause mortality and cardiovascular disease mortality.

Objective    To investigate the risk factors predicting the outcome of peritoneal dialysis (PD)-related bacterial-complicating peritonitis.    Methods    In this retrospective study, all the episodes of PD-related bacterial peritonitis presenting during Jan 2009 to Dec 2013 in our center were reviewed. Clinical and laboratory parameters at the onset of peritonitis, including patient demographic information, age, gender, duration of PD, residual renal function, local and systemic inflammation state, daily exchange number, peritoneal glucose exposure and so on, were recorded. Patients episodes were divided into three groups according to the outcome: complete cure (complete resolution of peritonitis without relapse or recurrence or repeat), peritonitis-related catheter removal/death group, and relapse   (relapse or recurrence or repeat) group.    Results    187 CAPD patients with 27.15(11.15, 53.13) PD duration were enrolled in the study. Total of 347 episodes of bacterial peritonitis in these patients were analyzed, with 130 episodes of gram-positive bacterial infection, 71 episodes of gram-negative bacterial infection, 15 episodes of polymicrobial and 131 episodes of cultured negative. Compared to the complete cure group and the relapse group, gram negative bacterial infection was more prevalent in the peritonitis-related catheter removal/death group. Furthermore, patients in the peritonitis-related catheter removal/death group showed longer PD age (P﹤0.01) and higher serum hs-CRP (P﹤0.01). Compared to the complete cure group, the serum albumin concentration was lower in the peritonitis-related catheter removal/death group (P﹤0.01). Kt/V was significantly lower in the relapse group than that in the complete cure group (P﹤0.05). Logistic analysis indicated age, non gram positive bacterial infection and increased hs-CRP were independent predictors for peritonitis-related catheter removal or death.    Conclusions    Age, non gram positive bacterial infection and hs-CRP are risk factors predicting peritonitis-related catheter removal or death in CAPD patients.

Objective    To identify the risk factors associated with cardiovascular and cerebrovascular disease (CCVD) in maintenance hemodialysis (MHD) patients.    Methods    We analyzed all of the patients undergoing maintenance hemodialysis in the dialysis center of the 3^rd Affiliated Hospital of Sun Yat-sen University for at least 3 months from Jan 1^st, 2009 to Dec 31^st, 2014. Baseline and yearly interval clinical data were recorded and patients were followed up until morbidity or death of CCVD. Cox proportional hazard regression and time-dependent Cox regression were used to estimate the relative risk of outcomes associated with clinical measurements.    Results    There were 243 patients enrolled in the study, with a mean age of (53.2±16.4) years old, and 138 of them were male (56.8%). The multivariate Cox proportional model revealed that age (HR=1.040, 95%CI: 1.015-1.065, P=0.002), Erythropoietin (EPO) dose (HR=0.914, 95%CI: 0.846-0.987, P=0.022) and history of cardiovascular and cerebrovascular disease (HR=4.045, 95%CI: 2.074-7.890, P＜0.001) were independent predictors of CCVD in MHD patients. After adjusting for baseline predictors, time-dependent serum phosphorus level (HR=1.722, 95%CI: 1.034-2.866, P=0.037) was significantly associated with CCVD.    Conclusion    Older age, decreases in EPO dose and history of cardiovascular and cerebrovascular disease were associated with increased risks of CCVD in MHD patients. Increase in serum phosphorus level was associated with increased risks of CCVD in a time-dependent manner.

Objective    To investigate the efficacy and safety of cutting balloon angioplasty for the treatment of hemodialysis arteriovenous fistula stenosis resistant to conventional percutaneous transluminal angioplasty (PTA).    Methods    The patients with arteriovenous fistula stenosis who had suboptimal results (residual stenosis ＞30%) by conventional PTA from December 2011 to February 2015 were enrolled. All the patients received cutting balloon angioplasty were rechecked every three months.    Results    A total of 25 patients with age of (60.7±12.9) years had suboptimal PTA results. Eleven patients with native arteriovenous fistula (AVF) and 14 patients with graft fistula (AVG) underwent cutting PTA for 30 times. The technical success rate was 86.7% and clinical success rate was 100%. The diameter stenosis pre-procedural and post-procedural of cutting PTA was (1.7±0.6) mm and (4.5±0.8) mm respectively (P＜0.05). Six patients had multiple lesions and the stenosis consisted of 21 outflow venous, 6 graft-to-vein anastomosis, 6 cephalic arch, 2 artery and 1 puncture hole stenosis. The primary access patency at 3 and 6 months for AVF group were 70.0% and 10.0%, while for AVG group the figures were 64.3% and 7.1% (P＞0.05). The secondary access patency at 3 and 6 months for AVF group were 70.0% and 30.0%, while for AVG group the figures were 85.7% and 64.3% (P＞0.05). The follow-up time was (8.1±7.3) months. The restenosis rate was 64.0%. Cutting PTA failed to achieve technical success for four times, of whom 2 patients required graft stent implantation and 2 patients required ultra-high-pressure balloons angioplasty to finally achieve technical success. The median survival time of fistula was 173 days.    Conclusions    Cutting balloon angioplasty have well short-term patency and safety in arteriovenous fistula stenosis resistant to conventional PTA, especially for calcified lesion or "balloon waist". Although it could provide a satisfied long patency by recurrent PTA, the use of cutting balloon would be not advocated as the first-line treatment for fistula stenosis. The efficacy superiority of cutting balloon between AVF and AVG, as well as the cost-effect comparison between cutting balloon and high-pressure balloon, remains unclear, the verification of which requires large-sampled, prospective and randomized studies.

Objective    To investigate the clinical-pathological features and prognosis of  idiopathic membranous nephropathy (IMN) with focal segmental lesion.    Methods    Two hundred and ninety-eight patients with biopsy-proven IMN in our hospital were retrospectively analyzed. The patients were divided into three groups: without focal segmental lesion group (FSL^-), with focal segmental glomerulosclerosis group (FSGS) and with early focal segmental lesion group (EFSL). The differences of clinical and pathological features and prognosis in the 3 groups were studied.    Results    There were later pathological stage, higher ratio of chronic renal tubulointerstitial damage and global glomerular sclerosis in FSGS group than those in the other two groups (all P＜0.05). The male ratio in EFSL group was higher than that in FSL^- group (P＜0.01), while the level of serum albumin was lower (P＜0.05). Compared with FSL^- group, there was longer average course before renal biopsy, higher blood pressure and levels of Scr in FSGS group (all P＜0.05)．Furthermore, the remission rate in EFSL group was lower than that in FSGS group and FSL^- group. Survival analysis showed that FSGS group had worse prognosis (FSGS to FSL^-, P=0.005, FSGS to EFSL, P=0.008). The analysis of risk factors suggested that triacylglycerol (OR=1.519, P=0.017), glomerulosclerosis (OR=1.073, P=0.041) and FSGS lesion (OR=5.960, P=0.009) were independent risk factors for renal death.    Conclusions    There were some differences between EFSL and FSGS lesion, both in clinical manifestations and pathology. FSGS lesion was independent predictive factor for progression to renal death. And the lowest remission rate was in EFSL group.

Objective    To explore whether angiopoietin-like protein 3 (Angptl3) is involved in the development of hyperlipidemia in nephrotic syndrome.    Methods    PCR analysis was carried out to identify the genotypes of Angptl3 Knockout mice. Sixty newborn Angptl3 knockout (KO) mice and wild type (WT) mice were randomly divided into four groups: KO-ADR, KO-Saline, WT-ADR and WT-Saline group. In each group  6 mice at different time points were separately analyzed: the pre-molding, molding 1st, 2nd, 4th, 8th week. WT-ADR and KO-ADR groups were treated with 25 mg/kg ADR once via tail vein injection at day 0; WT-saline and KO-saline groups were injected with the same volume of saline. Automatic biochemical analyzer was employed to test serum cholesterol (Cho) and triglycerides (TG) levels, ELISA method to detect the urine protein and urine creatinine, and real-time fluorescence quantitative PCR to detect the expression of Angptl3 mRNA in the renal tissue.    Results    (1)There were no significant differences in the weight, morphology or function of the liver and kidney between the KO and WT mice. Compared with WT mice, the levels of Cho and TG obviously decreased in the KO mice (P＜0.01). (2) In the WT-ADR group, urinary protein levels and the levels of Cho and TG increased significantly at 1st week after ADR injection (P＜0.05), while serum albumin level decreased dramatically (P＜0.05). The serum levels of Cho and TG increased gradually during the entire study period. The expressions of Angptl3 mRNA in kidney tissues were up-regulated significantly from 1st week (P＜0.05) to 8th weeks(P＜0.01). Furthermore, the expression of Angptl3  mRNA was significantly positively correlated with the Cho and TG levels (r=0.885, P＜0.01; r=0.788, P＜0.01, respectively). (3)The levels of Cho and TG in the Angptl3^-/- mice were lower than those in the WT mice during the entire study period after ADR injection (P＜0.05).    Conclusions    Angptl3 is involved in the development of hyperlipidemia in nephrotic syndrome. Knocking-out of Angptl3 may play an anti-dyslipidemic role in nephrotic syndrome.

Objective    To investigate the molecular mechanism of protection of ischemia preconditioning on renal ischemia reperfusion injury.    Methods    Male C57/BL6N mice were randomly divided into two groups: in IR group, 35 min ischemia was induced by occlusion of  both renal pedicles followed by 24 h perfusion (I/R). 15 min ischemia was induced 4 days before I/R in IPC group. Blood sample and kidney were collected in IR and IPC group after 24 h perfusion. Serum creatinine (Scr) and histological changes were used to evaluate the renal injury. PHD2 and HIF-1α were evaluated by Western blotting, miR-21 expression was confirmed by real-time PCR. In vitro, hypoxic model was established by 1% O₂ in HK-2 cells. Knockdown of miR-21 in hypoxic model was perfermed by locked nucleic acid modified-anti-miR-21 transfection. The levels of miR-21, HIF-1α and PHD2 mRNA were confirmed by real-time PCR. The levels of HIF-1α and PHD2 proteins were tested by Western blotting.    Results    In vivo, Compared with IR group, the renal function and histological changes were improved in IPC group (P＜0.01). Compared with IR group, the expression of miR-21(P＜0.01) and HIF-1α (P＜0.05) were increased in IPC group, while PHD2 was reduced (P＜0.01). In vitro, hypoxia reduced miR-21. The inhibition of miR-21 could increased the expression of PHD2 (P＜0.05).    Conclusions    Ischemia preconditioning may exert protection against renal ischemia reperfusion injury by inhibiting PHD2.

Objective    To observe the effect of ATF6 on the apoptosis and proliferation of podocytes induced by palmitic acid (PA).    Methods    Podocytes were stimulated with different doses of PA for 24 h. The expression of cleaved-caspase3 was detected by Western blotting. The podocyte apoptosis was analyzed by flow cytometry (FCM), and the expression of ATF6 was tested by Western blotting and immunofluorescence staining. After the transfection of adenovirus siRNA against ATF6, the proliferation, the cell cycle and apoptosis of potocytes stimulated with PA were tested by MTT or FCM.    Results    The levels of cleaved-caspase3 and ATF6 of podocytes stimulated with PA were significantly increased by a dose-dependent manner compared with the control group (P＜0.05). The apoptosis of podocytes stimulated with PA was increased (P＜0.05). Compared with the podocytes stimulated with PA, the apoptosis of podocytes transfected by adenovirus siRNA against ATF6 with PA stimulation was significantly reduced (P＜0.05). The proliferation of podocytes transfected by adenovirus siRNA against ATF6 and stimulated with PA, however, was obviously increased compared with the podocytes stimulated with PA (P＜0.05).    Conclusion    ATF6 mediated the apoptosis of podocytes induced by palmitate acid.

Objective    To investigate the effects and mechanisms of prostaglandin E₂ receptor subtype 3 (EP3) on transforming growth factor β1 (TGF-β1)-induced mouse mesangial cells damage.    Methods    Primary mouse mesangial cells were separated and cultured. Three siRNAs were synthesized and transfected into mesangial cells for silencing EP3 by Lipofectamine^TM 2000 and the best one was chosen. MCs were grouped into: (1)control group; (2)TGF-β1 (10 μg/L) group; (3)NC-siRNA plus TGF-β1 (10  μg/L) group; (4) EP3-siRNA group; (5)EP3-siRNA plus TGF-β1 (10 μg/L). Then the proliferation of MCs was evaluated by CCK-8 assay. The expression of PGE₂ and cAMP in cell supernatant were detected by ELISA. The mRNA and protein expression of fibronectin (FN), connective tissue growth factor (CTGF), cyclooxygenase-2 (COX2), membrane-bound prostaglandin E2 synthase 1 (mPGES1) were detected by real-time quantitative PCR and Western blotting. The phosphorylation of p38 MAPK and ERK1/2 was decected by Western blotting.    Results    Compared with control group, the cell proliferation  induced by TGF-β1 was increased (P＜0.05), the expression of PGE₂ and cAMP were improved, mRNA and protein expression of FN, CTGF, COX2 and mPGES1 were up-regulated (all P＜0.05). Compared with TGF-β1 group, the cell proliferation in EP3-siRNA plus TGF-β1 group was reduced, the expression of FN, CTGF, COX2 and mPGES1 mRNA and protein were downregulated (all P＜0.05), the phosphorylation of ERK1/2, p38 MAPK were also declined (P＜0.05).    Conclusion    EP3-siRNA may reduce TGF-β1-induced cell damage through upregulating the expression of cAMP, repressing the activity of ERK1/2 and p38 MAPK, inhibiting the expression of COX2 mPGES1 and PGE2 by feedback, then decreased the expression of FN and CTGF.

Objective    To explore the protective effects of adipose-derived stem cells (ADSCs) with phosphodiesterase 5 inhibition by lentivirus-mediated stable gene silencing on the proliferation and apoptosis of renal tubular epithelial cells induced by ischemia-reperfusion injury in vitro.    Methods    To isolate cultivate and indentify ADSCs from rats. Lentiviral expression vector of carrying PDE5 shRNA gene was transfected into ADSCs, and a negative control group was set up．Western blotting was used to detect PDE5 protein expression levels. ADSCs were co-cultured with NRK-52E in a transwell system, and NRK-52E cells were treated with ischemia/reoxygenation protocol. Edu assay was performed to evaluate the proliferation of NRK cells, flow cytometry to detect the apoptosis of NRK cells, and ELISA to quantify the protein expressions of fibroblast growth factor (FGF) and hepatocyte growth factor (HGF). The expression of E-cadherin and cytokeratin 18 (CK18) was quantified by real time PCR and flow cytometry.    Results    Western blotting for PDE5 protein indicated a significant reduction of PDE5 protein levels in PDE5 shRNA transduced population. After the treatment of ischemia/reoxygenation in vitro, the proliferative viability and apoptosis of NRK-52E cells co-cultured with ADSCs induced by PDE5 gene inhibition were significantly improved, compared to the normal group (all P＜0.05). And the release of HGF, FGF were markedly enhanced (all P＜0.05). Moreover, the NRK-52E cells survival, the expression of E-cadherin and CK18 on PDE5 inhibited ADSCs co-cultured with I/R injured NRK cells was significantly increased compared to that in the negative control group (all P＜0.05).    Conclusion    ADSCs preconditioned by inhibition of PDE5 can be a powerful novel approach to improve the survival of renal tubular cells following ischemia-reperfusion injury, and have an obvious tendency to transform epithelial cells.