Objective    To determine the condition of conjunctival and corneal calcificationin in maintenance haemodialysis patients, and explore the relationship between conjunctival and corneal calcificationin (CCC) and abdominal aortic calcification (AAC).    Methods     CCC was evaluated by slit-lamp eye photographs, and was graded and scored according to Porter’s classification system in the literature. Abdomen 1ateral X-ray examination were used as a criteria to determine the abdominal aortic calcification. The abdominal aortic calcification (AAC) score was calculated. Spearman correlation coefficient was used to analyze the relationship between CCC and AAC. Logistic regression analysis was used to determine the risk factor of CCC in MHD patients.    Results    Ninety-eight MHD patients were recruited. Their average age and dialysis vintage were (61.89±12.54) years and 51.67 (3.00～192.00) months, ninety-seven patients had conjunctival and corneal calcificationin, and seventy-two patients had abdominal aortic calcification, The median CCC was 8 (0, 17), the median AAC was 6 (0, 20), and the CCC was positively correlated with AAC (r=0.376, P＜0.001). Compared with the patients of CCC score≤5, the patients of CCC score＞10 had a higher level of phosphate, calcium-phosphate product,  iPTH, hs-CRP, and longer dialysis vintage, as well as a higher score of AAC (all P＜0.05). Multivariate logistic regression analysis showed that the higher score of AAC and the longer dialysis vintage were independent risk factors for severe corneal calcificationin calcification.    Conclusions    Conjunctival and corneal calcificationin is common in MHD patients, and CCC is positively associated with AAC, the risk of CCC rises as a result of a severer AAC and a longer dialysis vintage.

Objective    To evaluate the clinical features and the survival benefits of Chinese hemodialysis (HD) patients with central vein stenosis (CVS) undergoing different treatments.    Methods     From January 1, 2011 to Dec 31, 2012, 116 HD patients at high risk of CVS in Wuhan Central Hospital had their bilateral central veins assessed by vascular ultrasound and conventional venography. The clinical outcomes of 24 non-treated asymptomatic CVS patients, 17 non-treated symptomatic CVS patients and 6 treated symptomatic CVS patients were compared. Treatment costs of CVS were recorded and patients' survival rates were estimated by Kaplan-Meier analysis.    Results    Among 116 patients, 47 were diagnosed with CVS. The time span between symptomatic presentation and the diagnosis of CVS was more than 10 months averagely. Compared with non-CVS patients, the duration of HD in CVS patients was longer [(33.8±14.5) months vs (1.1±0.7) months, P﹤0.01] and the rate of central venous catheter (CVC) insertion was higher (87.2% vs 14.5%, P﹤0.01). Only 6 patients tried to maintain vascular access by endovascular intervention which costed ￥33 500 per person, much higher than other treatment options. While 30 patients refused endovascular intervention for fear of re-stenosis risk and high treatment costs, among whom 28 patients lost their initial vascular access. The 12-month survival rates of non-treated symptomatic CVS patients, treated symptomatic CVS patients and non-treated asymptomatic CVS patients were 84.6%, 88.9% and 87.0%, respectively, and the 24-month survival rates were 38.5%, 61% and 53.9%, respectively. No significant difference was found among the three groups.    Conclusions    Endovascular intervention may not be the first choice, but an alternative choice for Chinese HD patients with CVS, considering the long term survival benefit and the high treatment cost.

Objective    To observe the effect of ACEI/ARBs on peritoneal protein loss in peritoneal dialysis patients.    Methods    Total of 81 peritoneal dialysis patients were included in the study. Thirty-seven cases were treated with ACEI/ARBs(ACEI/ARBs group), forty-four cases did not receive any ACEI/ARBs (Control group). After 6 mouths, the effect of ACEI/ARBs on peritoneal protein loss was evaluated, and the effects of residual renal function and dialysis age on the peritoneal protein loss were statistically analyzed.    Results    (1) The peritoneal protein loss was reduced in ACEI/ARBs group, the difference was 1.2(0, 1.6) g/24 h, while in the control group, the protein loss had no statistically significant change, the difference between the two group was statistically significant (P＜0.05).  (2) When the patients’ eGFR＞2 ml•min^-1•(1.73 m²)^-1, the difference of the protein loss in ACEI/ARBs group was 1.4(1.2, 2.3) g/24 h , the difference between the two group was statistically significant(P＜0.01); when patients’ eGFR＜2 ml•min^-1•(1.73 m²)^-1, the differences of the protein loss between the two groups had no significant difference (P＞0.05). (3) When the dialysis ages＜12 months , the difference of the protein loss was 1.0(0.8, 1.4) g/24 h in ACEI/ARBs group，the differences between the two groups was statistically significant (P＜0.05); when the dialysis ages was from 12 months to 24 months or more than 24 months, The differences of the protein loss between the two groups and control group were both not statistically significant (P＞0.05).    Conclusion    ACEI/ARBs can reduce peritoneal protein loss in PD patient，the effect was better when patients’ residual renal function was better or dialysis age was shorter.

Objective    To investigate the association between nutrition and peripheral artery disease (PAD) in continuous ambulatory peritoneal dialysis (CAPD) patients.    Methods    One hundred and two stable CAPD patients from a single center were enrolled in this cross-sectional study. Automatic ankle-brachial index (ABI) measuring system was applied to examine ABI. Patients were divided into PAD group (ABI＜0.9) and non-PAD group (ABI≥0.9). Clinical data were collected. Biochemical parameters were detected. Nutritional status was evaluated by serum albumin, handgrip strength (HGS) and subjective global assessment (SGA). Logistic regression analysis was performed to estimate the association of PAD with each nutritional marker as well as other potential risk factors.     Results    The incidence of PAD was 23.53% (24/102). ABI was significantly lower in patients with malnutrition as compared to those without malnutrition [(0.72±0.21) vs (1.04±0.14), P＜0.01]. Compared with non-PAD patients, serum albumin (P＜0.01), HGS (P＜0.01), diastolic blood pressure (P＜0.05), serum creatine (P＜0.05)、blood urine nitrogen (P＜0.01) were significantly decreased, but age (P＜0.01), the incidence of malnutrition [SGA, P＜0.01], diabetic status (P＜0.01), cardiovascular disease history (P＜0.01) were significantly increased in PAD patients. Logistic regression analysis showed that serum albumin (OR=0.762, 95％CI：0.611-0.948, P=0.015), HGS (OR=0.988, 95％CI：0.979-0.997, P=0.013) were independent protective factors for PAD, malnutrition [(SGA), OR=21.101, 95％CI：5.008-88.901, P＜0.01] was independent risk factor for PAD in CAPD patients.    Conclusions    The PAD incidence of CAPD patients in our center is 23.53%. Nutrition is independent factor associated with PAD in CAPD patients.

Objective    To analysis the distribution and influence factors of N-terminal pro-brain natriuretic peptide (NT-pro BNP), and also its clinical significance though a cross-sectional survey of NT-pro BNP in maintenance hemodialysis patients in Zhongshan Hospital, Fudan University. Methods    A total of 207 stable hemodialysis patients were enrolled. The clinical parameters, plasma NT-proBNP levels and echocardiographic parameters were analyzed.    Results    Level of plasma NT-proBNP in patients with left ventricular hypertrophy (LVH) were significantly higher than those without LVH[M(1/4, 3/4): 3 104(1 626, 7 843) ng/L vs 1 291(772, 1 845) ng/L, P﹤0.01]. After logarithmic transformation for skewed variables NT-proBNP, log[NT-proBNP] was negatively correlated with hemoglobin (r=-0.212, P=0.004) and left ventricular ejection fraction (LVEF)(r=-0.202, P=0.003), and was positively correlated with left ventricular mass index (LVMI)(r=0.370, P=0.001), interdialysic weight gain (IDWG) rate (r=0.233, P=0.001), predialysis systolic blood pressure (r=0.345, P=0.001), predialysis diastolic blood pressure (r=0.152, P=0.032). The areas under curve(AUC) of NT-proBNP for diagnosing LVH and IDWG﹥4% were 0.786(95%CI 0.689-0.883, P﹤0.01) and 0.738(95%CI 0.667-0.810, P﹤0.01). When the threshold of NT-proBNP was set at 1 917 ng/L to diagnosis LVH, the sensitivity and specificity were 0.676 and 0.824. When the threshold of NT-proBNP was set at 2 872 ng/L to diagnosis IDWG﹥4%, the sensitivity and specificity were 0.704 and 0.758.    Conclusions    NT-proBNP levels are significantly abnormality in hemodialysis patients, mainly related with LVH, the high rate of IDWG, and the poorly controlled predialysis blood pressure. Proper dry weight assessment and strict control of IDWG may be effective way to intervene NT-proBNP.

Objective    To investigate the expression of CD26 (dipeptidyl peptidase 4) in the kidney tissues of diabetic rats and the effects of mycophenolate mofetil (MMF) on the renal CD26 expression.    Methods    Wistar rats were randomly divided into three groups: normal control group (NC group, n=7), diabetic model group (DM group, n=7) and MMF-treated group (MMF group, n=7). Wistar rats were fed with high-sucrose-high-fat diet and injected with streptozotocin into abdominal cavity to induce diabetes. Sixteen weeks later, blood glucose (BG), blood urea nitrogen (BUN), serum creatinine (Scr), renal hypertrophy index (kidney weight/body weight) and 24 hour urinary protein (24Upro) were measured. The number of CD3+/CD4+ T cells in renal tissues were measured through flow cytometry. The expression of CD26 in kidney was examined by using Western blotting and immunohistochemistry.    Results    Compared with NC group, BG, BUN, Scr, kidney weight/body weight, 24Upro were significantly increased in DM group (P﹤0.05). Except BG and kidney weight/body weight, the above-mentioned parameters were lower in MMF group compared with that in DM group (P﹤0.05). Intrarenal CD3+/CD4+ T cells were significantly up-regulated in DM group compared with that in NC group (P﹤0.01). CD26 in renal tissue was mainly expressed in T lymphocytes of renal interstitium. CD26 expression in DM group was significantly higher than that in NC group, and also higher than that in MMF group (P﹤0.05). In DM group, CD26+ T lymphocytes infiltration of renal interstitium was positively correlated with 24Upro (r2=0.770, P﹤0.05).    Conclusions    CD26 is related with diabetic nephropathy. MMF maybe inhibit T lymphocytes infiltration to reduce the expression of CD26 in renal interstitium, thus protecting the kidney function.

Objective    To explore the effects and mechanisms of prostaglandin E2 (PGE2) receptor 1 antagonist (SC-19220) on proliferation, prostaglandin synthase and extracellular regulated protein kinases (ERK) signal pathway induced by transforming growth factor β1(TGF-β1) in glomerular mesangial cells.    Methods    Mouse glomerular mesangial cells (GMCs) were divided into 5 groups: control group, TGF-β1 (10 μg/L) group, TGF-β1 (10 μg/L) plus SC-19220 group (0.1, 0.5, 1.0 μmol/L). The proliferation of GMCs was measured by CCK-8. The PGE2 in supernatant was measured by ELISA. The expression of connective tissue growth factor (CTGF), laminin (LN), cyclooxygenase 2(COX2), membrane-bound prostaglandin E2 synthase 1 (mPGES1) protein and mRNA was examined by Western blotting and real-time quantitative PCR, ERK1/2 or phospho-ERK1/2 was measured by Western blotting as well.    Results    TGF-β1 induced the proliferation of GMCs and increased the secretion of PGE2. Besides, TGF-β1 significantly up-regulated the expression of CTGF, LN, COX2 and mPGES1 mRNA and protein (P﹤0.05), and increased the expression of phospho-ERK1/2 protein (P﹤0.05). However, SC-19220 significantly attenuated the changes of above-mentioned parameters and their activities (P﹤0.05). All the effects of SC-19220 were in dose-dependent manner.    Conclusions    SC-19220 may reduce TGF-β1-induced cell damage by suppressing the activity of ERK1/2, and feedback inhibition of COX2, mPGES1 and PGE2, thus decreases the expression of  LN and CTGF.

Objective    To investigate whether the nod-like receptor (NLR) pathway is involved in protection of hydrogen sulfide (H2S) preconditioning during renal ischemia reperfusion.    Methods  Male Wistar rats were randomly divided into 3 groups: sham operation (Sham) group, renal ischemia/reperfusion (I/R) group subjected to occlusion of left renal pedicle for 45 min then reperfusion for 24 hours, and sodium hydrosulfide (NaHS) preconditioning group with continuous infusion of NaHS (300 nmol/min) by left renal artery for 15 min before I/R treatment. Renal injuries were evaluated by HE staining. The protein levels of NOD1, NOD2, nuclear NF-κB P65 and caspase-1 were analyzed by Western blot assay. The protein level of MCP-1 and IL-1β expressions was determined by immunohistochemical staining assay. Cell apoptosis were evaluated by Tunel staining assay.    Results     In I/R group, the renal NOD1 and NOD2 protein expressions were upregulated. Moreover, the nuclear NF-κB P65 expression was also elevated with an increase in its target genes-MCP-1 and IL-1β (All P＜0.01). HE staining revealed the existence of acute tubular necrosis in I/R kidney. TUNEL staining revealed more apoptotic cells in risk zone with the activation of caspase-1 of I/R-treated kidney(P＜0.01). NaHS preconditioning reversed I/R-induced increase in the expression of NOD1 and NOD2(P＜0.05). NaHS preconditioning also reduced I/R-induced activation of NF-κB P65 (P＜0.05) and upregulation of MCP-1 and IL-1β (P＜0.01). Moreover, NaHS preconditioning attenuated inflammation, repressed caspase-1 activation and reduced apoptotic cells after I/R.    Conclusion    Hydrogen sulfide preconditioning can alleviate renal ischemia/reperfusion injury by Nod-like receptor dependent on inflammatory pathway.

Objective    To observe the effect of irbesartan on the expression of angiopoietin-like protein 2 (ANGPTL2) in the diabetic rats kidney and explore the underlying mechanism.    Methods    A total of sixty male SD rats were divided into normal control group (NC group, n=15) and experimental group (n=45) randomly. The experimental group was fed with high sugar-fat diet and given a low dose streptozocin（STZ 30 mg/kg）to establish type 2 diabetic model. Rats successfully induced diabetes were randomly divided into 2 groups: diabetes group (DM) and irbesartan group (DI). Weight, blood pressure, blood glucose, serum creatinine (Scr), blood urea nitrogen(BUN), 24 hour urinary albumin(UAL) and renal histomorphology were observed after drug intervention at the 4th, 8th and 12th weeks. The expression of ANGPTL2 in renal tissue were detected by immunohistochemistry, real-time PCR and Western blotting.    Results    The levels of Scr, BUN, TG, TC and UAL in group DM were higher than in group NC at the 4th, 8th and 12th week (all P＜0.05). Compared with that in group DM, above indexes were lower in group DI at the 4th, 8th and 12th week (all P＜0.05). The pathological changes of the kidney in group DM were more serious than that in group DI. The expression of ANGPTL2 in group DM was much higher than that in group NC at the 4th, 8th and 12th week (all P＜0.05), and irbesartan treatment inhibited the up-regulation of ANGPTL2 in group DI(all P＜0.05).    Conclusion    The expression of ANGPTL2 increases in T2DM rats kidney tissue with time and irbesartan can inhibit the up-regulation of ANGPTL2 in T2DM rats.

Objective    To investigate the effects of pyrrolidine-dithio-carbamate ammonium (PDTC) on high-phosphate-induced vascular calcification in uremic rats.    Methods    Eight-week-old SD rats were pair-fed with standard chow containing 1.2% calcium and 0.6% phosphorus for the control group (n=8) or 0.75% adenine, 1.2% calcium, and 1% phosphorus for the chronic renal failure(CRF) group (n=8) or PDTC group (intraperitoneal injection, 100 mg•kg^-1•d^-1, n=8) for 8 weeks. The abdominal aortas were excised for Western blotting and immunostaining assay of NF-κB p65, osteopontin (OPN) and core binding factor α1(Cbfα1) protein.    Results    Serum urea nitrogen, creatinine, inorganic phosphate, calcium-phosphorus product increased significantly in CRF group and PDTC group after 4 weeks and 8 weeks (all P﹤0.01), although no differences were found between the latter two groups. After 8 weeks, aortic calcification was found in these two groups, immunostaining assay revealed OPN and Cbfα1 expressed in aortic intima, media and adventitia, and Western blotting analysis showed that total NF-κB p65, nuclear phosphorylated-p65 (p-p65), OPN and Cbfα1 expressions were significantly higher than those in control group (all P﹤0.01). The expression of total p65 and p-p65 was positively correlated with Cbfα1(r=0.707, P﹤0.01; r=0.507, P﹤0.01).  Conclusion  PDTC alleviates inorganic phosphate-induced aortic calcification significantly by inhibiting the nuclear translocation of NF-κB p65 and the expression of Cbfα1.

Objective    To investigate the effect of Notch signaling during bone marrow mesenchymal stem cells (BMSC) differentiating into islet in vitro.    Methods    The specific inhibitor of γ-secretase DAPT was used to inhibit the Notch  signaling pathway. After induction, DTZ staining, indirect immunofluorescence staining, RT-PCR and Western blotting were used to detect the expression of insulin, glucagon, Pdx-1 and Ngn3.    Results    (1) Identification of BMSCs: Indirect immunofluorescence staining showed that BMSCs could express CD59 and CD90, which both were makrers of mesenchymal stem cells. Besides, BMSCs could express nerve culluar markers such as NSE, GFAP, suggesting multi-directional differentiation. (2) The result of MTT showed DAPT could inhibit the cell proliferation in a time-dependent manner and a dose-dependent mannar. Besides, DAPT could inhibit the expression of target gene of Notch signal pathway in a time-dependent manner and a dose-dependent mannar. After treated by 1, 5, 20 μmol DAPT, the expression of Hes1 had reached to 92.06%, 71.40% and 46.89% of controls respectively, suggesting efficiency of inhibition on Notch reached 7.94%, 28.6% and 53.11% respectively (all P＜0.05). (3) Indirect immunofluorescence staining showed the expression of pancreas-specific markers such as insulin and glucagon were much higher in DAPT treated BMSCs than that in controls, which was confirmed by RT-PCR and Western blotting analyses. The proportion of insulin-producing cells differentiated from DAPT treated BMSCs was (74.03±3.96)%, which was higher than that from controls[(36.49±3.24)%, P＜0.05]. (4) Furthermore, RT-PCR and Western blotting analysis showed that the expressions of Pdx-1 and Ngn3 were earlier than that of insulin and glucagon, and the expressions of Pdx-1 and Ngn3 were higher in DAPT treated BMSCs than that in controls.    Conclusions    Notch signaling pathway plays a role in the differentiation of BMSCs into islet in vitro. Pharmacological interference with Notch signaling pathway may provide a novel method to obtain islet for therapeutic use.