Objective To analyze the clinic-pathological data and peritubular capillary (PTC) injuries of malignant nephrosclerosis (MN) patients and their correlations with the long term renal survival. Methods This was a retrospective cohort study of 52 MN patients in Peking Union Medical College Hospital from January 2003 to March 2012. Their clinical data and renal biopsy samples were carefully studied. CD34 staining was performed to evaluate the PTC area, using Benign nephrosclerosis (BN, n=17) patients and glomerular minimal lesions (GML, n=19) patients as controls. Multivariate Cox proportional hazard model was used to identify the potential independent risk factors for long term renal survival. Results Fifty-two MN patients were enrolled. The sex ratio of male to female was 12∶1 and the average age was (34.0±8.2) years. The maximum blood pressure (SBP/DBP) was (230.4±25.0)/(156.4±20.6) mmHg, companied with significant loss of eGFR and proteinuria. Glomerular sclerosis index, tubular atrophy and interstitial fibrosis correlated with eGFR and proteinuria(P＜0.05). After aggressive treatment, BP control rate improved significantly (76.9% vs 3.7%, P＜0.01), Scr [(376.4±263.8) μmol/L vs (486.8±375.7) μmol/L, Wilcoxon test, P＜0.01] and proteinuria [(1.10±0.70) g/24 h vs (2.04±1.26) g/24 h, P＜0.01, n=21] also improved. PTC area in MN patients was significantly lower than those in BN patients and GML patients, and it correlated well with Scr (r=-0.553, P=0.001) and eGFR (r=0.476, P=0.004). The median follow-up time was 74 months, the cumulative renal survival rate at 1 year, 5 year and 10 year was 90%, 64% and 23%, respectively. Kaplan-Meier analysis showed that the patients with higher PTC area had longer renal survival time [(114.8±12.4) months vs (63.0±8.3) months, χ2=5.312, P＜0.05]. Univariate Cox proportional hazard model found that unsatisfied BP control, eGFR＜30 ml?min-1?(1.73 m2)-1 upon discharge, lower PTC area, severer tubular-interstitial damage and anemia were associated with poor renal outcome. Multivariate Cox model showed that unsatisfied BP control (RR=3.89, 95% CI 1.75-8.65, P=0.001), eGFR＜30 ml?min-1?(1.73 m2)-1 upon discharge (RR=4.27, 95%CI 1.40-13.09, P=0.011) were independent risk factors for long-term renal survival. Conclusions The correlation between PTC area and renal functions in MN patients are much better than that of classic vascular changes. Unsatisfied BP control and eGFR＜30 ml?min-1?(1.73 m2)-1 upon discharge are independent risk factors for long-term renal survival.

Objective To investigate the relationship between the expression of Wnt induced secreted protein-1 (WISP-1) and the fibrosis of renal biopsy tissue in IgA nephropathy (IgAN) patients. Methods Fifty-three patients firstly diagnosed as IgA nephropathy by renal biopsy were included and classified according to Oxford and Lee's classification. Sixteen patients with MCD entered the fibrosis negative control group, and fourteen healthy adults entered the normal control group. The expression of WISP-1 in renal tissues and serum of all subjects were detected by immunohistochemistry and ELISA respectively. Results Immunohistochemistry results showed that WISP-1 was not expressed in MCD patients and normal human kidney tissues, which was abundantly deposited in renal tissue of patients with focal proliferative IgAN with renal interstitial fibrosis. The serum level of WISP-1 in IgAN patients was significantly higher than that in normal subjects (P=0.015) and MCD patients (P=0.030). In the subgroup analysis of IgAN renal fibrosis, the serum concentration of WISP-1 of fibrosis grade between 0-10% (F1 group) and fibrosis＞25% (F3 group) were significantly higher than that in the normal group and the MCD group (all P＜0.05). There was no significant difference between F2 group (10%＜fibrosis≤25%) and normal group or MCD group (P＞0.05). Conclusions The expression of WISP-1 in serum and renal tissue of renal interstitial fibrosis IgAN patients is higher than that of normal and MCD patients without renal fibrosis, and the IgAN patients' serum level of WISP-1 is significantly increased in fibrosis lower score group. The expressions of WISP-1 in serum and renal tissue are related to the occurrence of IgAN renal interstitial fibrosis, in which WISP-1 may play an important role as an early precursor factor in the pathogenesis of IgAN renal interstitial fibrosis.

Objective To analyze the related factors of micro-albuminuria and macro-albuminuria in type 1 diabetes mellitus (DM) in the classification tree model, and to screen the high risk population of diabetic kidney disease. Methods 394 patients with type 1 diabetes were enrolled in the hospital from 2008 to 2015. According to glomerular filtration rates and urine albumin quantification, the patients were divided into type 1 diabetes group (299 cases), micro-albuminuria group (73 cases) and macro-albuminuria group (22 cases). The classification tree model was utilized to analyze related factors of the different stages of proteinuria, and the high-risk population was screened by node gain analysis. Results Four important explanatory variables were screened out by the classification tree model from the 27 candidate variables related to primary renal damage, including retinopathy, fibrinogen, waist-hip ratio (WHR), red blood cell distribution width (RDW). Retinopathy was an major factor of DKD. The probability of macro-albuminuria in retinopathy and WHR＞0.82 group was 43.8%, and if at the same time RDW＞0.14, the probability of macro-albuminuria was 88.9%. Conclusions The classification tree model can analyze factors of the separate stages of proteinuria in type 1 diabetic patients effectively. Retinopathy is the major influential factors of type 1 diabetic patients with proteinuria.

Objective To study the incidence and risk factors of new foot ulcer among diabetic patients on peritoneal dialysis. Methods This is a single-center prospective cohort study. Clinically-stable diabetic patients on peritoneal dialysis in our renal division were recruited from January 2014 to June 2014. Baseline data including general information, biochemistry data, dialysis adequacy, the dorsalis pedis artery pulse, clinical symptoms of diabetic foot and ankle brachial index were recorded. All patients were followed till to Dec 31, 2015. The outcomes consisted of new foot ulcer, amputation due to foot ulcer or gangrene, and lower limb vascular blood supply revascularization. Results Totally 108 patients were recruited and followed up the average time (17.7±5.6) months. Among 108 patients, 16 cases had a history of diabetic foot ulcer, and 1 case had amputation. During the follow-up, 11 cases (10.2%) had new foot ulcer, 3 cases (2.8%) had amputation due to foot ulcers or gangrene, and 8 cases (7.4%) had lower limb vascular blood supply revascularization. A total of 13 cases (12%) had composite end points with 81.3 times/1000 patients of incidence. Univariate and multivariate Cox regression models showed that the history of foot ulcer was the only independent risk factors for new foot ulcers-related composite end points. Conclusion The incidence of new foot ulcer-related composite end points was 12%, which could be independently predicted by the history of diabetic foot ulcer.

Objective To explore the effect and safety of mycophenolate mofetil (MMF) and glucocorticoid on Henoch-Schonlein purpura nephritis in children and compared with cyclophosphamide (CTX). Methods The data of 48 patients diagnosed as Henoch-Schonlein purpura by renal biopsy were retrospectively analyzed. Median follow-up time was 22(7, 48) months. The subjects were divided into 2 groups. 34 cases were in the MMF group: MMF (15-20 mg?kg-1?d-1 or 800-1200 mg/m2)+ prednisone, and 14 cases in the CTX group: CTX (8 - 12 mg?kg-1?d-1)+prednisone. Clinical and laboratory data were collected at baseline and 1, 3, 6 months after treatment. During follow - up, cumulative retreatment rate and adverse reactions after treatment were recorded. Results In two groups after treatment for 1, 3, 6 months, 24 hours urinary protein quantitative was significantly lower than the baseline value, serum albumin (sAlb) was significantly higher than the baseline value, and serum creatinine (Scr) indicated no statistically significant difference during the follow-up period. After the treatment of 1 month, the efficient rate of MMF group was higher than the CTX group (MMF 73.5 % vs 42.9%, P=0.046), the effective treatment of 3, 6 months at the end of the follow-up, no statistically significant difference were observed in the accumulative remission rate. The total rate of retreatment was 10.4% (5/48), where MMF group was lower that of the than CTX group (3.0% vs 28.6%, P＜ 0.001). The retreatment often occurred after discontinuation of prednisone and CTX, MMF reduction process. Eleven children received IMPDH2 gene polymorphisms test in MMF group, 9 AA children had shorter time for drugs to be effective than that of the AG and GG children. The incidence of adverse reactions of MMF group was obviously lower than CTX group at the end of the follow-up (8.8% vs 35.7%, P=0.025), where two groups developed fungal infection. Conclusions The short-term effect of both groups are the same, but the recurrent rate and incidence of adverse reactions of MMF group are lower than those of the CTX group. The preliminary study shows that IMPDH2 gene polymorphisms is associated with MMF curative effect and adverse reactions.

Objective To explore the association of fibroblast growth factor-23 (FGF23) with abdominal aortic calcification(AAC) and adverse outcomes in maintenance hemodialysis patients. Methods One hundred and fourteen cases of MHD patients were collected prospectively. Serum intact FGF23 was detected by ELISA. Abdomen lateral plain was used as a criteria to determine the abdominal aortic calcification and the abdominal aortic calcification score was counted. Logistic regression analysis was used to determine the risk factors of AAC. Kaplan-Meier analysis was applied to compare the survival rate among different groups and COX regression analysis was used to determine the association of FGF23 and mortality in MHD patients. Results Seventy-six patients present abdominal aortic calcification. The median of AACS was 4.0(0.0, 11.0). The median level of FGF23 was 7277.4(2535.0, 9990.8) pg/ml. The median follow-up duration was 72.0(67.8, 72.8) months. During the follow-up, 22 patients (19.3%) died of all-cause death and 17 cases (14.9%) died of cardiovascular diseases. Serum FGF23 level was positively correlated with AAC (r=0.285, P=0.002). Logistic regression analysis showed that longer age (OR=1.059, 95%CI: 1.020-1.100, P=0.003) and dialysis vintage (OR=1.009, 95%CI 1.000-1.017, P=0.039), smoking history (OR=3.010, 95%CI 1.177-7.696, P=0.021) and higher FGF23 level(OR=2.831, 95%CI 1.010-7.937, P=0.048) were independent risk factors of moderate to severe AAC in MHD patients. Kaplan-Meier survival curves showed that the patients with AACS≥5 had significantly higher all-cause mortality(P=0.028) and CVD mortality (P=0.035) than those with AACS＜5. However, the Kaplan-Meier analysis showed no significant difference regarding the level of serum FGF23 with the all-cause and CVD mortality. Cox regression demonstrated that FGF23 was not associated with increased mortality risk, neither in crude nor in multivariate adjusted models. Conclusions Abdominal aortic calcification had a high prevalence in MHD patients. The all-cause and CVD mortality was higher in patients with moderate to severe AAC. FGF23 was an independent risk factor of moderate to severe AAC, but it can't yet be a predictor for the all-cause and CVD mortality of MHD patients.

Objective To discuss the efficacy and safety of transversus abdominis plane block (TAPB) combined with rectus sheath block (RSB) for peritoneal dialysis catheter placement. Methods Thirty patients, ASAⅠorⅡ, body mass index (BMI) 18-30 kg/m2, were scheduled for elective peritoneal dialysis catheter placement. They were randomly divided into three groups: local anesthesia group (group L), ultrasound-guided TAPB group (group T), ultrasound-guided TAPB combined with RSB group (group TR). Mean arterial blood pressure (MAP) and heart rate (HR) were recorded before (T0) and after (T1) anaesthesia, the beginning of operation (T2) and abdomen-closing (T3), 6 h (T4), 12 h (T5), 24 h (T6) after surgery. The consumption of local anesthetic during the surgery, dezocine after the surgery, the score of visual analogue scale (VAS) during rest and movement at T2-T6 and the adverse reactions were recorded. Results At T3, MAP and HR in group L and group T were higher than those in group TR (P＜0.05). At T4 and T5, MAP and HR in group L were higher than those in group T and group TR (P＜0.05); the consumption of local anesthetic ropivacaine in group L and group T were more than that in group TR (P＜0.05); the consumption of local anesthetic ropivacaine and dezocine in group L were more than that in group T (P＜0.05). The VAS score was significantly lower in group T and group TR at T3, T4, T5 compared with that in group L (P＜0.05). There were no adverse reactions. Conclusion Ultrasound-guided TAPB combined with RSB is safe and efficacious in patients undergoing peritoneal dialysis catheter placement.

Objective To investigate the role of the group IB secretory phospholipase A2 (sPLA2-IB) and M-type phospholipase A2 receptor (PLA2R) in human podocyte injury and its possible signal transduction pathway. Methods Differentiated human podocytes were exposed to PBS or different sPLA2-IB concentration conditions (10-9, 10-7, 10-5 mol/L) for 2 hours. The wound healing assay was used to measure cell migration rate; Apoptosis in cultured human podocytes was assessed by Hoechst 33342 staining and flow cytometry; Western blot was used to analyze the protein expression of p-cPLA2α, p-p38, and p53. Then control siRNA or PLA2R siRNA were transfected to podocytes. Podocytes were divided into normal control group, negative control siRNA group and PLA2R siRNA group. Twenty four hous later, the cells were stimulated by 10-5 mol/L sPLA2-IB for 2 hours. The protein expression of p-cPLA2α, p-p38, and p53 were detected by Western blot. Results Compared to PBS control group, the migration ability of podocytes decreased when stimulated with sPLA2-IB (10-7 mol/L-10-5 mol/L), and the apoptosis of podocytes increased in a concentration-dependent manner, the protein level of p-cPLA2α, p-p38 and p53 protein increased too. After the knockdown of PLA2R by PLA2R siRNA transfection, stimulated the podocytes with the same dosage of sPLA2-IB, the protein expression of p-cPLA2α, p-p38 and p53 all decreased. Conclusion sPLA2-IB stimulation can increase human podocyte apoptosis and decrease its migration ability. The possible mechanism might be through p38-cPLA2α-p53 pathway.

Objective To investigate the effect of klotho on the human vein umbilical endothelial cells (HUVECs) injury induced by indoxyl sulfate (IS) and to explore its mechanism and the role of endoplasmic reticulum stress (ERS) in this process. Methods (1) The cell vitalities of HUVECs incubated with different concentration of IS (5, 25, 50 mg/L) for 48 h and with 50 mg/L IS for different time points (12, 24, 48 h) were measured by CCK-8 assay. (2) HUVECs were incubated with 50 mg/L IS and different concentration of klotho (0, 1, 10, 100 μg/L) for 48 h and their cell viabilities were measured by CCK-8 assay. (3) HUVECs were divided into four groups: control group, IS group (50 mg/L IS), klotho group (50 mg/L IS+100 μg/L klotho) and Compound C group (50 mg/L IS+100 μg/L klotho+10 μmol/L Compound C). The cell vitality and the apoptosis of HUVECs were evaluated by CCK-8 assay and flow cytometry, respectively. The mRNA and protein expressions of GRP78 and CHOP were measured by real-time PCR and Western blotting. The phosphorylation level of AMPK was tested by Western blotting. Results IS inhibited cell vitality in the time-dependent and concentration-dependent manner. The cell viability of HUVECs with 50 mg/L IS was lower than normal control (P＜0.05). The inhibited cell vitality induced by IS was partly restored by klotho in concentration-dependent manner. The cell viability was higher in 100 μg/L klotho+50 mg/L IS group than 50 mg/L IS group (P＜0.05). Compared with control group, the expressions of GRP78 and CHOP and cell apoptosis increased, however, the level of phosphorylated AMPK (p-AMPK) decreased in IS group (all P＜0.05). Compared with IS group, the expressions of GRP78 and CHOP and cell apoptosis decreased and the level of p-AMPK increased in klotho group (all P＜0.05). Furthermore, the above effects of klotho could be partly blocked by Compound C. The above indexes showed statistical differences between Compound C group and klotho group. Conclusions IS can inhibit the HUVECs cell vitality, and induce ERS and cell apoptosis. Klotho protein could antagonize the above effects, probably through activating AMPK pathway and reducing ERS-mediated cell apoptosis.

Objective To observe the expression of cysteine-rich protein 61 (Cyr61) in transforming growth factor -β1 (TGF-β1)-activated renal fibroblasts (NRK-49F), and to explore its effect and mechanism. Methods (1) NRK-49F cells were activated by TGF-β1 with different concentrations (0.0, 0.5, 1.0, 2.0, 5.0 μg/L). Western blotting was used to detect the expression of Cyr61 protein, and CCK-8 assay was used to test the proliferative activity of NRK-49F cells. (2) NRK-49F cells with low expression and over expression of Cyr61 were established by plasmid transfection. The cells were divided into control group (null vector transfection), over-expression group and low-expression group. The proliferation was discovered by CCK-8 assay after 24, 48 and 72 h. Further, 5.0 μg/L TGF-β1 activated these three groups. The proliferation was also discovered by CCK-8 assay and the cell cycle was analyzed by flow cytometry. The mRNA expressions of fibrosis markers (Col1α1, Col3α1, MMP9, MMP13) and factors of cell senescence signal pathway (p53, p21, Rb, p16) were ascertained by real time PCR, and the protein expressions of Col3 and MMP9 were detected by Western blotting. Results (1) Compared with 0.0 μg/L TGF-β1 group, the proliferation of NRK-49F cells was enhanced in 0.5, 1.0, 2.0 and 5.0 μg/L TGF-β1 groups (all P＜0.05), while the expression of Cyr61 protein was decreased in 1.0 μg/L group and increased in 5.0 μg/L group (all P＜0.05). (2) The proliferation of over-expression group was lower than that of control group after 24, 48 and 72 h (all P＜0.05), which was in a time-dependent manner. (3) Compared with control group activated by TGF-β1, the over-expression group expressed less fibrosis factors (Col1α1 and Col3α1) and more anti-fibrosis factors (MMP9 and MMP13) with decreased proliferation (all P＜0.05). Simultaneously, the proportion of cells bogged down in G1 phases, as well as the expressions of p53, p21 and Rb mRNA increased (all P＜0.05). The above effects of low-expression group were just opposite to over-expression group. Moreover, there was no significant difference in the expression of p16 gene among the three groups (P＞0.05). Conclusions Cyr61 can curb the proliferation and fibrotic phenotypes of fibroblasts, thereafter slowing down the process of renal fibrosis. The p53/p21/Rb interrelated cell senescence signal pathway may be involved in the anti-fibrosis process.

Objective To investigate the role of STAT3 transcription factor in IL-6 inducing epithelial mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs). Methods HPMCs were cultured in vitro and grouped. (1) According to the stimulation time with 50 μg/L IL-6, HPMCs were divided into 24, 48, 72 h groups. (2) HPMCs were grouped 50, 100 μg/L according to IL-6 concentration. (3) HPMCs were respectively divided into control group, IL-6 group, empty vector group, empty vector+IL-6 group, virus infecting group and virus infecting+IL-6 group, as lenti-virus vector mediating RNA interference targeting STAT3 was applied. The mRNA expressions of E-cadherin, α-smooth muscle actin (α-SMA) and vascular endothelial growth factor (VEGF) were detected by real time PCR; their protein expressions and the phosphorylation of JAK2 and STAT3 were detected by Western blotting; the expressions and distribution of E-cadherin and α-SMA were observed by immunofluorescence. Results Compared with those in control group, the expression of E-cadherin decreased remarkably (P＜0.05), while the expressions of VEGF and α-SMA and the ratio of phosphorylated (p)-JAK2/JAK2 and p-STAT3/STAT3 increased significantly in IL-6 concentration groups and stimulation time groups (all P＜0.05), which had been dose and time dependent. Compared with empty vector+IL-6 group, virus infecting+IL-6 group had decreased expressions of VEGF and α-SMA, while increased expressions of E-cadherin (all P＜0.05). Conclusions IL-6 can promote VEGF and α-SMA gene expression and prevent E-cadherin gene expression by STAT3, which involves in EMT of peritoneum fibrosis. While STAT3 gene is knocked-down, EMT is inhibited in HPMCs.