Objective To clarify the role of pentraxin 3 (PTX3) in the development of peripheral arterial disease (PAD) in maintenance hemodialysis (MHD) patients. Methods One hundred and sixteen patients undergone MHD therapy in our center for more than 3 months were enrolled in the study. Clinical data were collected for analysis. Ankle-brachial index (ABI) was used to estimate the presence of PAD. Patients were divided into PAD group (ABI<0.9) and non-PAD group (ABI≥0.9). Plasma PTX3 was measured by ELISA. Logistic regression was used to estimate the association of PAD with PTX3 as well as other potential risk factors. Results The incidence of PAD was 18.1% (21/116). Plasma level of PTX3 was significantly higher in PAD patients than that in non-PAD patients [(5.55±2.63) μg/L vs (2.32±1.29) μg/L, P<0.01]. Univariate analysis showed ABI values were negatively correlated with plasma PTX3 levels (r=-0.548, P＜0.01), high-sensitivity C-reactive protein (hsCRP), age, blood glucose and triglyceride. ROC curve of PAD revealed that area under curve (AUC) of PTX3 was 0.901 (P＜0.01). With the cut-off value of PTX3 as 4.06 μg/L, the diagnostic sensitivity and specificity in PAD were 81.0% and 91.5%. ROC curve of PAD showed that AUC of hsCRP was 0.640 (P＜0.05). With the cut-off value of hsCRP as 3.33 mg/L, the diagnostic sensitivity and specificity in PAD were 57.1% and 56.8%. Using Logistic regression, plasma PTX3 was found to be associated with PAD (OR=9.755, 95%CI:2.359-19.354, P=0.001). Conclusions The PAD incidence of MHD patients in our center is 18.1%. Plasma PTX3 level is significantly correlated with the presence of PAD in MHD patients. The sensitivity and specificity of PTX3 are higher than those of hsCRP for PAD diagnosis.

Objective To investigate the situation of prevalence, treatment and control of hypertension in patients with chronic kidney disease (CKD) by cross-sectional study. Methods Nine hundred out-patients with CKD in our department from November 2006 to March 2007 were enrolled in the study, including 480 male and 420 female. Among 900 CKD cases, 354 patients underwent maintenance dialysis, including 228 on hemodialysis and 126 on peritoneal dialysis. Results The prevalence of hypertension in CKD patients was 80.2%（male 83.5% vs female 76.4%, P<0.01）. The prevalence of hypertension in patients on dialysis was significantly higher than that in non-dialysis patients (90.1% vs 73.8%, P<0.01), but there was no significant difference between hemodialysis and peritoneal dialysis cases. Antihypertensive treatment rate was 92.4% in CKD patients with hypertension, and was significantly higher in patients on dialysis than that in non-dialysis patients（95.6% vs 89.8%, P<0.01）. The control rate according to current recommendations for CKD patients（BP<130/80 mm Hg） was very low. Control of both SBP and DBP was only achieved in 20.4% of non- dialysis patients. The control rate of hypertension（BP<125/75 mm Hg） in patients with proteinuria >1 g/24 h was 8.4%. The proportion of dialysis patients with BP<140/90 mm Hg was significantly lower than that of non-dialysis patients (45.2% vs 55.5%, P<0.01）. The percentage of hemodialysis patients with BP<140/90 mm Hg was significantly higher than that of peritoneal dialysis patients (49.8% vs 36.5%, P<0.05）. The prevalence of hypertension was associated with the decrease of renal function and the increase of age. The prevalence of hypertension in diabetic nephropathy was higher than that in primary glomerular diseases. Patients received 1, 2, 3 and 4 or more kinds of antihypertensive drugs accounted for 37.2%, 37.5%, 19.3% and 5.9% respectively. The combination of calcium channel blocker (CCB) and renin-angiotensin-aldosterone system (RAAS) inhibitors was more frequently used in CKD patients. The CCB was the most frequently prescribed drug (74.1%), followed by angiotensin II receptor blockers (ARB) (48.4%), angiotensin-converting enzyme inhibitors (ACEI) (25.6%) and alpha, beta-blockers(24.7%). Conclusions The prevalence of hypertension in CKD patients is quite high, which is associated with the progression of renal function, increase of age, the type of underlying kidney disease, obesity and diabetes mellitus. The control of hypertension is unsatisfied in CKD patients, especially in dialysis patients and those with overt proteinuria.

Objective To investigate the safety and tolerability of darbepoetin alfa, a long-lasting erythropoietin, single intravenous administration in maintenance hemodialysis (MHD) patients. Methods A single center, open clinical trial was carried out. Forty-three stable MHD patients were divided into 5 groups and received darbepoetin α at dosage of 0.225, 0.45, 0.9, 1.8, 3.6 μg/kg respectively. The vital signs, symptoms, ECG and laboratory examinations were monitored and detected before and after administration. Results Of the 43 patients (male 26 and female 17), the largest tolerable dosage of darbepoetin alfa was 3.6 μg/kg. During the study, the main side effect associated with darbepoetin was hypertension aggravation (7%). One patient died but that was not associated with darbepoetin alfa. Conclusion Darbepoetin alfa is safe and well tolerated.

Objective To investigate the influence of a combination of Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis (A&A) on the production and elimination of reactive oxygen species (ROS) and the underlying mechanism during the process of renal interstitial fibrosis in the obstructive kidneys of rats with unilateral ureteral obstruction (UUO). Methods Male Wistar rats were randomly divided into control, sham, UUO and UAA (UUO+A&A) groups. The rats in UAA group were administered with A&A (14 g/kg) by oral gavage once daily; the ones in sham and UUO group were given with equal volumes of water. Three days after setting up models, pathological injury of renal tissue was evaluated. Level of total antioxidant capacity (T-AOC) and activity of CuZn superoxide dismutase (CuZn-SOD) in renal homogenates were measured by spectrophotometry. Expression of NADPH oxidase subunits p47-phox, p22-phox and nitrotyrosine was analyzed by Western blot. Results Severe interstitial inflammatory cells infiltration, mild tubular atrophy and interstitial fibrosis were found in UUO group, which were alleviated by A&A administration. Compared to sham group, T-AOC of UUO group was not significantly changed, but the expression of the nitrotyrosine, NADPH oxidase subunits p47-phox and p22-phox was increased significantly (P<0.05). After A&A administration, T-AOC level was increased (2.5±1.1 vs 1.5±0.5, P<0.05) and the expression of nitrotyrosine was decreased (P<0.05) in UAA group compared with UUO group. Additionally, the expression of p47-phox was reduced at day 3 (P<0.05), accompanied with a reduced expression of p22-phox (P< 0.05). CuZn-SOD activity was not significantly changed among the groups. Conclusion The inhibition of A&A in NADPH oxidase subunits p47-phox and p22-phox, which is responsible for reduction of oxidative stress, is associated with the alleviation of renal fibrosis in obstructive rat kidney.

Objective To investigate the influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells in diabetic rats and to explore the mechanism. Methods Wistar rats were randomly divided into three groups: control, diabetes and fasudil-treatment. All the rats were sacrificed after three months of feeding with or without fasudil. Pathological changes of the glomeruli and renal interstitium were studied by periodic acid-Schiff’s staining and Masson staining, respectively. Expression of ROCKI, α-SMA, E-cadherin and the distribution of β-catenin was examined by immunohistochemistry. Changes in the MYPT1 phosphorylation profile and α-SMA, E-cadherin and membrane β-catenin expression were detected by Western blot. Changes in the levels of ROCKI, E-cadherin and total β-catenin mRNA expression were analyzed by real-time PCR. Results Fasudil treatment notably attenuated renal interstitial fibrosis in diabetic rats. Compared to the control rats, diabetic rats showed an elevated phosphorylation of MYPT1 (P<0.01）, increased expression of α-SMA (P<0.01), decreased expression of E-cadherin and membrane β-catenin (P<0.01, respectively) and increased expression of ROCKI, total β-catenin mRNA(P<0.01, respectively）, decreased expression of E-cadherin mRNA（P<0.01）. Fasudil treatment for diabetic rats attenuated MYPT1 phosphorylation (P<0.01）, decreased α-SMA expression（P<0.01), increased E-cadherin and membrane β-catenin expression （P<0.01, respectively), and reduced ROCKI, total β-catenin mRNA expression (P<0.01, respectively), increased expression of E-cadherin mRNA (P<0.01). Conclusions Fasudil may reduce the epithelial-mesenchymal transdifferentiation and renal interstitial fibrosis in diabetic rats through the inhibition of ROCK activity. Such effect further facilitates the recovery of the cell-cell adhesion among renal tubular epithelial cells and the formation of adhesion complex.

Objective To investigate the effect of different concentrations of rapamycin on the proliferation and apoptosis of glomerular mesangial cells (GMCs) and to investigate the mechanism. Methods GMCs were treated with different concentrations of rapamycin(1 μg/L, 2 μg/L, 4 μg/L, 8 μg/L, 16 μg/L). After treatment for 24 h, 48 h and 72 h, cell proliferation was assessed by MTT colorimetric assay and the growth curve was traced. After treatment for 72 h, the cell cycle distribution and the apoptotic rate of GMCs in different concentrations of rapamycin were analyzed by flow cytometry. The effects of different concentrations of rapamycin on the mRNA and protein expression of p27 and p53 were detected by RT-PCR and Western blot respectively. Result The low dose of rapamycin(1 μg/L) could significantly inhibit the proliferation of GMCs and showed no effect on apoptosis. The high dose of rapamycin(8-16 μg/L) could significantly increase the apoptotic rate of GMCs. Rapamycin could increase the mRNA and protein expression of p27 and p53. Conclusion Rapamycin can inhibit GMCs proliferation and promote GMCs apoptosis by increasing the expression of p27 and p53.

Objective To explore the impact of high glucose on the expression of ubiquitin mRNA and ubiquitinated-protein in cultured rat glomerular mesangial cells(GMCs). Methods Cultured HBZY-1 rat GMCs were divided into 5 groups: normal glucose group (5.6 mmol/L), high glucose group(10, 20, 30 mmol/L). Mannitol group was used as high osmosis control. The expression of ubiquitinated-protein and ubiquitin mRNA of each group was measured by Western blot and real-time PCR respectively. Results Ubiquitinated-protein was mostly macromolecule. Compared with normal glucose group, the expression of ubiquitinated-protein was increased significantly in a dose- and time-dependent manner in 20,30 mmol/L glucose groups (all P＜0.01), and the expression of ubiquitin mRNA was also increased obviously (P＜0.05). Conclusions High glucose can obviously increase the expression of ubiquitinated-protein and ubiquitin mRNA, and upregulate ubiquitin-proteasome pathway in cultured GMCs

Objective To observe the effect of 1，25(OH)2D3 on expression of vitamin D receptor (VDR), tumor necrosis factor α (TNF-α) and transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells (RPMCs) stimulated by lipopolysaccharide(LPS), so as to provide some evidence for clinical use of 1，25(OH)2D3 on peritoneal dialysis-associated peritoneal inflammation. Methods RPMCs were isolated, cultured and passaged by enzymatic disaggregation, then identified by phase contrast inverted microscope, transmission electron microscope with immunocytochemistry method. RPMCs were incubated with LPS (1，10，100 mg/L)and LPS (10 mg/L) for 2,6,12 hours, or stimulated by 1，25（OH）2D3 (10-8 mol/L, 10-7 mol/L, 10-6 mol/L) after incubated with LPS (10 mg/L) for 2 hours. RPMCs in the control group were just incubated with medium. Expression of VDR mRNA and protein was detected by RT-PCR and Western blot. In addition, ELISA was performed to investigate the changes of TNF-α and TGF-β1 in culture medium. Results Compared with control group, the expression of VDR mRNA and protein was decreased in LPS group (P<0．05). LPS could significantly induce the expression of TNF-α and TGF-β1 in RPMC(P<0．01), which could be partially reversed by different concentrations of 1，25（OH）2D3 (P<0．01). Conclusions 1，25（OH）2D3 can reverse the decrease of VDR mRNA and protein induced by LPS as well as the induction of TNF-α and TGF-β1 up-regulated by LPS in RPMC in a dose- and time-dependent manner. Our research provides experimental evidence of VDR ameliorating peritoneal dialysis-associated peritoneal inflammation and peritoneal fibrosis.

Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms’ tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol(50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.