Objective To investigate the expression of endostatin (ES) in rat peritoneum and its association with peritoneal neoangiogensis. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal control rats (C group), renal failure without PD rats (non-PD group), rats dialyzed with 1.5% PD solution (1.5% PD group) and 4.25% PD solution (4.25% PD group). After regular PD for 28 days, mRNA and protein expression of ES in peritoneal tissues of each group were detected by RT-PCR and immunohistochemistry respectively. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results ES mRNA was expressed in each group, 0.47±0.05 in C group, 0.45±0.04 in non-PD group, 0.46±0.04 in 1.5%PD group, 0.47±0.03 in 4.25%PD group, and no significant differences were found among groups. Score of ES protein expression was O in C group, 2 in non-PD group, 4 in 1.5%PD group, and 9 in 4.25%PD group. MVD was 3.13±1.13 in C group, 5.13±1.14 in non-PD group, 9.00±1.51 in 1.5%PD group, 10.75±1.83 in 4.25%PD group, and significant differences were found among groups. Conclusion Uremia circumstance and non-physiological compatibility peritoneal dialysate can increase ES protein expression and MVD, which may participate in and have effects on the course of peritoneal neoangiogensis.

Objective To observe the clinical curative effect and side-effect of MMF plus corticosteroid hormone compared with cyclophosphamide(CTX)plus corticosteroid hormone for idio-focal segmental glomerular sclerosis(I-FSGS).Methods Thirty patients with I-FSGS confirmed by renal biopsy in the Nephrology Department,the First Affiliated Hospital of Zhengzhou University from 2004 to 2006 were randomly divided into two groups-15 patients in each group:MMF combined with corticosteroid hormone as therapy group,and CTX combined with corticosteroid hormone as control group.The two groups were compared on urine protein in 24 hours,serum albumin and creatinine clearance rate(Ccr).Results There were significant differences(P

Objective To investigate the effects of the cyclooxygenase-2 (COX-2) inhibitor (celecoxib) on angiogenesis and peritoneal function of uremic peritoneal dialysis rats.Methods Forty-eight male SD rats were selected,and they were randomly divided into five groups:normal control group(n =8),sham operation group(n =8),uremia group(5/6 nephrectomy,n =8),PD group [4.25％ PD solution,2 weeks PD model(n =8) and 4 weeks PD model(n =8)],PD + celecoxib intervention group[treated by celecoxib(20 mg/kg) via oral gavage,n =8].The peritoneum of uremic peritoneal dialysis rats was observed in different dialysis time from peritoneal structures,functions,peritoneal tissue capillary density (microvessel density,MVD) and COX-2,vascular endothelial growth factor (VEGF) expression level,and the impacts of celecoxib on uremic peritoneal dialysis rats peritoneal angiogenesis and peritoneal function were study.Results With the conduct of the peritoneal dialysis,peritoneal thickness increased,the inflammatory cells infiltrated,peritoneal equilibration test (PET) showed that ultrafiltration volume decreased significantly (P ＜ 0.05),the amount of glucose transport rate rised significantly (P ＜ 0.05),but the celecoxib could improve net ultrafiltration volume (P ＜ 0.05),and reduce the glucose transport rate (P ＜ 0.05).The peritoneal tissue MVD and COX-2,VEGF expression were significantly increased in uremia group and PD group compared with that in the normal control group (all P ＜ 0.05),were significantly lower in PD + Celecoxib intervention group than that in uremia group (P ＜ 0.05).The correlation analysis showed that the level of COX-2 protein expression with MVD,VEGF protein expression was positively correlated (both P ＜ 0.05),the level of VEGF protein expression and MVD was positively correlated (P ＜ 0.05).Conclusions In vivo high glucose dialysate and uremia environmental can stimulate the COX-2 and VEGF expression raised,and the capillaries production increased in peritoneal tissue.Celecoxib can alleviate the change of peritoneal tissue morphology and function in long-term peritoneal dialysis rats.Celecoxib inhibits the peritoneal neovascularization of uremic peritoneal dialysis rats,possibly through inhibition of COX-2 expression to reduce the production of VEGF.

Objective To investigate the effects of cyclooxygenase-2 (COX-2) inhibitor on peritoneal lymphangiogenesis and peritoneum function in uremic rat.Methods Uremic rats treated by peritoneal dialysis were intragastric administration celecoxib.Structures of peritoneum,peritoneal function,peritoneal lymphatic vessel density (LVD) were detected in every group.The mRNA of vascular endothelial growth factor-C (VEGF-C),lymphatic vessel endothelial hyluronan receptor-1 (LYVE-1) and COX-2 were tested by RT-PCR.The protein expressions of LYVE-1,VEGF-C,COX-2 were tested by western blot.Results With the extension of the duration of dialysis,the peritoneum thickness was increasing,inflammatory cell infiltrated obviously,uhrafiltration volume decreased significantly.But the celecoxib could increase uhrafiltration volume and reduce the glucose transport rate(P ＜0.05).Compared with the normal group,the levels of LVD,COX-2,VEGF-C,and LYVE-1 mRNA and protein were significantly up-regulated in uremic and dialysis groups (P ＜0.05).Compared with the uremic dialysis group,the levels of LVD,COX-2,VEGFC and LYVE-1 mRNA and protein were significantly down-regulated in the celecoxib group.There was a positive correlation between COX-2 and VEGF-C,LVD in protein levels,as well as VEGF-C and LVD (all P values ＜ 0.05).Conclusions Hyper glucose dialysis solution and uremic condition could up-regulate the expression of COX-2,VEGF-C,LYVE-1 in gene and protein level and stimulate lymphangiogenesis.COX-2 inhibitor could delay the change of peritoneal structures and function.COX-2 inhibitor could prevem the lymphangiogenesis in uremic rat treated by peritoneal dialysis,which might down-regulate the expression of VEGF-C by COX-2 depended manner.

Objective To study the effect of recombinant human edostatin on peritoneal angiogenesis in uremic peritoneal dialysis(PD) rats. Methods Forty male SD rats were randomly divided into 5 groups: normal control rats (group 1), renal failure without PD rats (group 2), rats dialyzed with 4.25% PD solution (group 3), rats dialyzed with 4.25% PD solution and received subcutaneous injection of recombinant human endostatin 10 mg/kg (group 4), rats dialyzed with 4.25% PD solution and received subcutaneous injection of recombinant human endostatin 40 mg/kg (group 5). Recombinant human endostatin was given every other day during peritoneal dialysis period, total 14 times. After regular PD for 28 days, tissue immunohistochemical staining and RT-PCR were used to detect the mRNA and protein expressions of VEGF and bFGF in peritoneal tissues of each group rats. Microvessel density (MVD) of peritoneum was detected and quantified with anti-CD34 immunohistochemical staining. Results The mRNA and protein of VEGF and bFGF were expressed in each group. Compared to group 1, the mRNA and protein expression of VEGF and bFGF were significantly up-regulated in group 2 and group 3 (all P＜0.05). Compared with group 3, the mRNA and protein expression of VEGF and bFGF were significantly downregulated in group 4 and group 5 (all P＜0.05). Compared with group 4, the mRNA and protein expression of VEGF and bFGF were significantly down-regulated in group 5 (all P＜0.05). The new microvascular vessels in group 1 showed little or none. Compared with group 1, MVD was significantly increased in group 2 and group 3 (P＜0.05). Compared with group 3, MVD was significantly decreased in group 4 and group 5 (all P＜0.05). Conclusions Recombinant human endostatin can effectively inhibit rat peritoneal neoangiogensis. Down-regulated expression of VEGF and bFGF in peritoneum may be one of the mechanisms of recombinant human endostatin inhibiting peritoneal angiogenesis.

Objective To investigate the efficacy and feasibility of combination of leflunomide and clopi-dogrel in the treatment of IgA nephropathy. Methods 84 patients with primary IgA nephropathy were randomly divided into four groups: Control group (group 1) received valsartan; test group (group 2) received valsartan +clopidogrel; group 3 received valsartan + leflunomide; group 4 received valsartan + clopidogrel + leflunomide. The level of 24 h urinary protein,serum creatinine, glomerular filtration rate and therapeutic efficacy, adverse reactions were detected and recorded during the 48 weeks follow-up. Results The 24 h urinary protein of 84 patients showed a downward trend during the course of treatment , but there were statistical differences between groups since 8 weeks after treatment (P < 0.05). The level of urine protein in group 3 and group 4 decreased significantly to 61.48% and 67.23% respectively in 48 weeks after treatment. At the end of the follow up , the serum creatinine levels in group 1 and group 2 significantly increased while the glomerular filtration rate de-creased obviously (P < 0.05). Conclusion The combination of leflunomide with clopidogrel could reduce the level of urinary proteins and slow renal function deterioration in the treatment of IgA nephropathy with less ad-verse reactions. This study provides a new idea for treatment of IgA nephropathy.

Objective To observe the role and related mechanism of chemerin and its receptor ChemR23 in glomerular endothelial cells (GEnCs) stimulated by high glucose.Methods Mouse GEnCs were cultured and divided into control group,20.0 mmol/L high glucose group,40.0 mmol/L high glucose group and mannitol control group.Then the expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cell culture supematant as well as the expressions of intracellular protein and mRNA of chemerin,ChemR23,IL-6 and TNF-α were detected.Lentiviral transfection targeting ChemR23 was applied before high glucose-or Chemerin-stimulated,and expressions of supernatant and intracellular mRNA of IL-6 and TNF-α were measured.Meanwhile whether p38 mitogen-activated protein kinase (p38 MAPK) pathway was activated by high glucose was detected.The specific inhibitor of p38 MAPK was added prior to high glucose-stimulated,then supernatant and intracellular mRNA expressions of IL-6 and TNF-α was detected.The supernatant expressions of IL-6 and TNF-α were measured by ELISA.The intracellular protein expression and p38 MAPK phosphorylation activity were detected by Western blotting.The mRNA expression was detected by real time PCR.Results Compared with those in the control group,in high glucose groups the expressions of IL-6,TNF-α and chemerin were significantly increased (all P ＜ 0.05),however,the expressions of ChemR23 did not change (all P ＞ 0.05);the supernatant and mRNA expressions of IL-6 and TNF-α were also elevated in the chemerin group (all P ＜ 0.05).Lentivirus baring shRNA could efficiently suppress ChemR23 expression,and the Chemerin-or high glucose-induced expressions of IL-6 and TNF-α were reduced (all P ＜ 0.05).Also it could significantly reduce the expression of phosphorylated-p38 MAPK (p-p38 MAPK) induced by high glucose (P ＜ 0.05),as high glucose group had higher p-p38 MAPK than control group (P ＜ 0.05).While the high glucose-elevated expressions of IL-6 and TNF-α were significantly attenuated by p38 MAPK inhibitor (all P ＜ 0.05).Conclusions High glucose stimulation can induce the expression of chemerin in GEnCs.By binding to ChemR23,chemerin activates p38 MAPK signaling pathway,and then promotes the expressions of IL-6 and TNF-α.These inflammatory cytokines aggravate inflammation of GEnCs.

Objective To investigate the effect of soluble Tie2 fusion protein(sTie2/Fc)on the angiogenesis of peritoneal vessels in uremic peritoneal dialysis (PD) rats.Methods Rats were randomly divided into 6 groups:normal rats as control group (group1),rats with sham operation (group2),uremic rats without PD (group3),uremic rats dialyzed with 4:25％ PD solution (group4),uremic rats dialyzed with 4.25％ PD solution and treated by subcutaneous injection of 2.5 μg/kg sTie2/Fc (group5),uremic rats dialyzed with 4.25％ PD solution and treated by subcutaneous injection of 5.0 μg/kg sTie2/Fc (group6).sTie2/Fc was given every other day during peritoneal dialysis period,total 14 doses.After regular PD for 28 days,RT-PCR and tissue immunohistochemical staining were used to detect the mRNA and protein expressions of Angpt-2 in peritoneal tissues in each group of rats.Microvessel density (MVD) of peritoneum was detected and quantified with immunohistochemical staining by using anti-CD34 antibody.Results The expression of Angpt-2 mRNA and protein was found in each group.There was no significant difference of.Angpt-2 expression both in mRNA and protein level between group1 and group2.Compared with group1,the mRNA and protein expression of Angpt-2 were significantly increased in group3 and group4 (all P＜0.05).Compared with group3,the mRNA and protein expression of Angpt-2 were significantly increased in group4 (all P＜0.05).Compared with group 4,the mRNA and protein expression of Angpt-2 were significantly decreased in group5 and group6 (all P＜0.05).Compared with group5,the mRNA and protein expression of Angpt-2 were significantly decreased in group6 (all P＜0.05).Only few new microvessel was found in group1 and group2.Compared with group1,MVD was significantly up-regulated in group3 and group4 (all P＜0.05).Compared with group4,MVD was significantly down-regulated in group5 and group6 (all P＜0.05).Conclusions Peritoneum neoangiogensis can be effectively inhibited by sTie2/Fc in uremic rat treated with PD.Blocking of signal transduction may be involved in the mechanism of sTie2/Fc inhibiting peritoneal angiogenesis.

Objective:To investigate changes in serum C3d and C5b-9 levels in elderly patients with idiopathic membranous nephropathy(IMN)and their correlations with prognosis.Methods:Two hundred thirty-one elderly patients with IMN and 96 non-elderly patients with IMN confirmed by kidney biopsy at the First Affiliated Hospital of Zhengzhou University from January 2015 to May 2017 were enrolled.During the same period, 118 healthy individuals receiving health checkups were included as controls.Patients were divided into the low C3d group( n=112)and the high C3d group( n=113)according to the median level of serum C3d.Serum C3d and C5b-9 levels were measured by enzyme-linked immunosorbent assays. Results:Serum C3d and C5b-9 levels in elderly IMN patients were 0.23(0.15, 0.45)mg/L and 0.28(0.20, 1.23)mg/L, respectively, which were higher than those in healthy controls[0.18(0.13, 0.22)mg/L, 0.22(0.16, 0.26)mg/L, respectively]( Z=-4.261 and -6.213, P<0.001). Serum C3d levels in elderly and non-elderly IMN patients were correlated negatively with the estimated glomerular filtration rate( r=-0.155 and -0.426, P=0.019 and 0.000), but positively with serum creatinine, anti-phospholipase A2 receptor(PLA2R)antibody levels and 24 h urinary protein( r=0.184, 0.326, 0.407, 0.321 and 0.145, P=0.005, 0.001, 0.000, 0.001 and 0.027). Kaplan-Meier survival analysis showed that the cumulative renal survival rate in elderly IMN patients was lower in the high C3d group than in the low C3d group(47.8% vs.70.8%, Log Rank χ2=7.399, P=0.007). Multivariate Cox regression analysis showed that high C3d levels were an independent risk factor for poor renal outcomes in elderly IMN patients( HR=2.288, 95% CI: 1.082-4.839, P=0.030). Conclusions:High serum C3d levels are associated with increases in urinary protein excretion and anti-PLA2R antibody levels, renal function decline, and poor renal outcomes in elderly IMN patients.