Objective To evaluate the sensitivity and specificity of plain radiography in the diagnosis of vascular calcification in maintenance hemodialysis (MHD) patients. Methods Multi-slice computed tomography (MSCT) was used as the reference standard in the assessment of vascular calcification in MHD patients. A total of 54 MHD patients, 26 male and 28 female, mean age (60.4±13.3) years, underwent both MSCT and plain radiography of lateral abdomen and pelvis to evaluate abdominal aortic calcification, bilateral iliac and femoral artery calcification. Abdominal aorta was divided into upper and lower segment by L2-L3 intervertebral space. The severity of vascular calcification by MSCT was graded from score 0 to 5. Two independent radiologists analyzed the results of plain radiography and MSCT, and inter-observer agreements were calculated by using κ statistics. Results According to the results of MSCT, the calcification rate of abdominal aorta was 86.1%, and the calcification rate of iliac and femoral artery was 74.5%. There was significant difference of the calcification rate between large artery and muscular arteries. Inter-observer agreement of calcification was excellent (κ=0.864-0.893). Compared with MSCT, the specificity of plain radiography with regard to detection of abdominal aortic, iliac and femoral calcification were 100%. The sensitivity of plain radiography was different according to the different MSCT score, which was as follows: MSCT score≥grade 1: 60.2% and 24.8% for lateral abdomen radiography to detect abdominal aortic calcification and pelvic radiography to detect iliac, femoral calcification respectively; MSCT score≥ grade 2: 76.9% and 43.5% respectively; MSCT score≥grade 3: 100% and 74.4% respectively. Conclusions The sensitivity of plain radiography in the assessment of vascular calcification increases with the severity of calcification. The sensitivity in the assessment of abdominal aortic calcification is higher than that of iliac and femoral artery calcification. Plain radiography can be used to detect moderate to severe vascular calcification in MHD patients.

Objective To investigate the incidence of acute kidney injury ( AKI) post-orthotopic liver transplant ( OLT ) and its association with prognosis. Methods Data of 28 patients received single OLT in our hospital from 2004 to 2006 were retrospectively analyzed. The incidence of AKI was investigated by new acute kidney injury network (AKIN) criteria. The follow-up was over one year. The prognosis of AKI patients at day 28 and 1 year was evaluated by Kaplan-Meier survival analysis. The association between AKI and prognosis was examined. Results A total of 193 patients were enrolled. The average age was (48.07±10.02) years old. The ratio of male to female was 4:1. One hundred and sixteen (60.1%) patients of post-OLT AKI were found, whose AKI stage 1, 2 and 3 were 50.0%, 21.6% and 28.4% respectively. Ten (8.6%) patients required renal replacement therapy (RRT) after OLT. In AKI post-OLT patients, day 28 and 1 year mortality were significantly higher than those in non-AKI patients (15.5% vs 0, 25.9% vs 3.9%, respectively, both P<0.05). Kaplan-Meier survival analysis showed the 1-year survival rates of AKI stage 1, 2, 3 post-OLT and non-AKI were 84.0%, 81.0%, 42.4% and 90.9%, respectively. The 1-year survival rate of non-AKI was significantly higher than that of AKI stage 1, 2, 3. The 1-year survival rate of AKI stage 3 was significantly lower than that of stage 1 and 2. There was no significant difference between AKI stage 1 and 2. Scr at 1 year post-OLT was significantly higher than that of baseline [（88.35±37.15） vs (73.70±33.88) ?滋mol/L， P<0.05）. The change of Scr value at 1 year compared to baseline in AKI patients was similar to non-AKI patients. However such change in AKI stage 2 and 3 was higher than that in stage 1. Conclusions The incidence of AKI post-OLT is quite high and associated to the poor prognosis in short and long periods. Renal function may decrease gradually which is associated to the AKI stage post-OLTI.

Objective To investigate the prevalence，clinical features and prognosis of pulmonary arterial hypertension (PAH) in maintenance hemodialysis (MHD) patients for early diagnosis and treatment. Methods Complete clinical data of 184 MHD patients in Beijing Chaoyang Hospital between January 2000 and December 2007 were retrospectively analyzed. PAH diagnosis depended on echocardiography. Results Sixty-five (35.3% ) patients were found having PAH, including 31 females and 34 males, with mean (56.84±14.58) years old. The mean hemodialysis duration was (29.69±21.61) months. Among 65 patients with PAH, arteriovenous fistula was used in 61 patients and central venous catheter in 4 patients. The systolic pulmonary arterial pressure (SPAP) was (44.56±8.25) mm Hg(1 mm Hg=0.133 kPa) in PAH group, which was significantly higher than (30.28±3.92) mm Hg in non-PAH group. There were significant differences of interval dialysis weight gain (IDWG), Hb, Hct, right atrial diameter, pulmonary artery diameter and right ventricular diameter between PAH group and non-PAH group (all P<0.05)． Multivariate analysis revealed anemia, IDWG and right atrial diameter were independent factors (P<0.05). There were no significant differences of age, dialysis duration, serum calcium, phosphorus, alkaline phosphatase, parathyroid hormone (PTH) between two groups. Conclusions PAH is a common complication of MHD patients. Patients with mild PAH had few clinical manifestations and good therapeutic responses, but severe PAH may be associated with poor prognosis. Doppler echocardiography should be performed periodically to detect PAH in MHD patients.

Objective To report a Chinese boy suffering from nephrotic syndrome associated with Schimke immuno-osseous dysplasia（SIOD）. Methods The clnical data and pathological changes of renal biopsy were analyzed and associated literatures were reviewed. The clinicopathological features and diagnosis of SIOD were discussed. Results The first symptom of the patient was recurrent infections. Growth retardation, spondyloepiphyseal dysplasia accompanied by nephrotic syndrome and defective cellular immunity were seen as clinical features in this patient. Renal pathology showed focal segmental glomerulosclerosis. Conclusion Combining the clinical manifestation with renal pathology, the case is diagnosed as Schimke immuno-osseous dysplasia.

Objective To compare the efficacy of continuous ambulatory peritoneal dialysis (CAPD) and hemodialysis (HD) on polycystic kidney disease (PKD) patients with end-stage renal disease (ESRD). Methods Retrospective analysis was made on 29 patients with PKD who carried out dialysis therapy for over 3 months in our department from January 2001 to December 2007. They were divided into the CAPD group (10 cases, 34.5%) and HD group (19 cases, 65.5%). Ten cases of non-PKD CAPD patients were randomly selected as the control, who matched the CAPD group in terms of age and gender. The patient information was recorded, such as general data, initial dialysis data, complications, survival time, quit of dialysis or death, etc. Kaplan-Meier method and Log-rank test were adopted to analyze the survival rate. Results The survival rates of 1-, 3- and 5-year for the CAPD group were 90％, 75％ and 25％ respectively, while for the HD group were 94.4％, 67.6％, and 48.3％, and for the control were 83.3％, 44.4％ and 22.2％ respectively, with no significant differences among 3 groups (P>0.05). Kt/V[（2.09±0.97）/W vs (1.93±0.59)/W] and Ccr [（58.5±9.1） L&#8226;W-1&#8226;(1.73 m2)-1 vs (55.0±9.5) L&#8226;W-1&#8226;(1.73 m2)-1] of the first appraisal of the sufficiency of peritoneal dialysis for the CAPD group and the control were quite similar. The incidence of peritonitis for the CAPD group (0.62 times/patient year) was similar to that for the control (0.30 times/patient year)（P>0.05）. The duration of the first peritonitis[(23.5±4.0) months vs (20.0±15.8) months] and the catheter exit-site infection (0 time vs 1 time) for two groups were similar as well （P>0.05）. One patient had hernia in CAPD group and no patient in control group had hernia. The incidence of peritoneal dialysate leakage was similar between these two groups. In the HD group, two patients (10.5%) had cerebral hemorrhage resulting in death, and 10 patients (52.6%) had cystic hemorrhage, 5 out of whom underwent operation due to repeated cystic hemorrhage and 2 cases received unilateral nephrectomy because of severe hemorrhage. No patient in CAPD group had cerebral hemorrhage but 1 patient (10%) had cystic hemorrhage and recovered after conservative treatment. The hemorrhage complication incidence of CAPD group was significantly lower than that of HD group（P<0.05）. Conclusions The prognosis and complication incidence in PKD and non-PKD patients treated with CAPD are similar. The prognosis of PKD patients treated with CAPD or HD is also similar, and the risk of hemorrhage complications of PKD patients treated with CAPD may be decreased compared with those treated with HD. PKD patients can choose HD or PD as the initial therapy of ESRD unless existence of hernia or intolerance. PKD is not the contraindication of PD.

Objective To investigate the influence of heme oxygenase-1 (HO-1) on rat renal tubular epithelial cell apoptosis induced by albumin and the possible mechanism. Methods The renal tubular epithelial cells(NRK-52E) were cultured in DMEM/F12 1:1 medium as normal control group; NRK-52E cells were cultured with 30 g/L fat-free bovine serum albumin(BSA) as the BSA control group; NRK-52E cells were cultured with CoPP (Cobalt protoporphyrin IX) 5 μmol/L for 24 hours as the treatment group． MTT assay was used to observe the effects of CoPP on growth inhibition induced by BSA in NRK-52E cells. The effect of CoPP was observed in BSA-induced apoptosis with the fluorescence microscope dyed by AnnexinV-FITC PI. The levels of HO-1, and expression of Bcl-2 and Bax mRNA were detected by reverse transcript polymerase chain reaction (RT-PCR). Results Compared with normal control group, BSA inhibited the growth of NRK-52E cells（P<0.05) and increased cell apoptosis rate（P<0.05). The CoPP pretreatment partially inhibited the BSA-induced apoptosis（P<0.05). Compared with normal control group, HO-1 mRNA expression increased（0.44±0.06 vs 0.39±0.05, P<0.05） in BSA control group. Compared with the BSA control group, the expression of HO-1 mRNA significantly increased after CoPP pretreatment（0.50±0.06 vs 0.44±0.06, P<0.05）. Meanwhile, BSA increased the expression of Bax mRNA （0.87±0.04 vs 0.67±0.03, P<0.05）and reduced the expression of Bcl-2 mRNA（0.25±0.04 vs 0.42±0.02, P<0.05）. CoPP could inhibit the effect of BSA （Bax mRNA: 0.75±0.07, Bcl-2 mRNA: 0.36±0.03, P<0.05, respectively）. Conclusions BSA can increase the apoptosis rate significantly and regulate the expression of apoptosis associated proteins in mRNA level directly. CoPP inhibits these changes, which provides evidence to support the essential role of HO-1 in cytoprotective function .

Objective To investigate if Rac1 GTPase activation plays an important role in hyperpermeability and tyrosine phosphorylation of tight junction induced by vascular endothelial growth factor (VEGF) in glomerular endothelial cells(GEnCs). Methods Primary cultured rat endothelial cells were used as experimental model. The effect of VEGF at different concentrations (5 or 50 μg/L) on endothelial permeability was investigated by transendothelial electrical resistance (TEER). The permeability of GEnCs transfected with wild type Rac1 (wtRac1) or dominant negative Rac1 (N17Rac1) was also detected. Immune precipitation and immune blotting were used to detect the tyrosine phosphor-occludin in GEnCs. Results VEGF at high concentration (50 μg/L) induced hyperpermeability in GEnCs (P<0.05). At the same time, GTP-binding and membrane-bound Rac1 GTPase significantly increased(P<0.01)in GEnCs. Tyrosine phosphor-occludin was also increased(P<0.05) under VEGF stimulation. However, transfection of GEnCs with N17Rac1 dramatically attenuated the effect of VEGF on tyrosine phospho-occludin and endothelial cell permeability. Conclusions Increased VEGF can induce hyperpermeability in glomerular endothelial cells, which is related to occludin tyrosine phosphorylation through Rac1 activation. It provides a framework for understanding the role of VEGF-induced Rac1-phospho-occludin pathway in the integrity of barrier function in the diabetic milieu.

Objective To observe the steps of vascular smooth muscle cells(VSMCs) calcification induced by high phosphate enviroment in vitro. Methods VSMCs were incubated with high phosphate(2.5 mmol/L or 3.5 mmol/L) medium for different times. Expression of core binding factor α1(Cbfα1), osteopontin(OP), collagen type I(Col I), osteocalcin(OC) and α-smooth muscle actin(α-SMA) was investigated by Western blot, immunofluorescence staining and real time PCR. Mineral deposition was assessed by von Kossa and Alizarin red staining. Ultrastructure of VSMCs calcification was observed by electron microscopy (EM). Results Up-regulated expression of osteoblast-specific transcription factor Cbfα1 in the nuclei occured at as early as 12 hours. The protein of Col I and OP was up-regulated when VSMCs were incubated in high phosphate medium for 3 days, and content of OC increased at the time of 6 days. When cultured in 2.5 mmol/L phosphate medium for 15 days, VSMCs lost their lineage marker α-SMA, developed granular calcium deposits. Moreover, the results of real time PCR indicated mRNA level of OP and Col I increased at day 1, OC increased at day 5 and α-SMA level decreased at day 10, respectively. Ultrastructural analysis also confirmed the presence of collagen and matrix vesicles in the cells. Conclusion VSMCs phenotype transformation induced by high phosphate enviroment is an orchestrated, highly regulated process.

Objective To investigate the interference and associated mechanism of human tissue kallikrein (HK) gene on renal interstitial fibrosis in rats with 5/6 nephrectomy. Methods Human kallikrein cDNA was packed in a recombinant adeno-associated virus(rAAV)-based plasmid vector. The rAAV-HK was produced by transfection in 293 cells. Twenty-four male Wistar rats were divided into sham operation and operation groups. The rats with 5/6 nephrectomy were randomly divided into simple operation, control and experiment groups. The rats in experiment group received single dose rAAV-HK via the tail vein with 1×1011 pfu. Before nephrectomy and every month after surgery until the rats were sacrificed, the caudal arterial pressure was measured using tail cuff blood pressure determinator. Three months after HK gene delivery, the rats were sacrificed. The expression of HK in rats was assessed by RT-PCR，Western blot and enzyme-linked immunosorbent assay(ELISA). The pathological changes of renal interstitium were evaluated by Masson stainning, and the distribution of bradykinin B2 receptor (BKB2R) and angiotensinⅡtype1 receptor(AT1R) was examined by immunohistochemistry. The expressions of BKB2R, AT1R, p-MAPK protein in renal tissue were detected by Western blot. Results Three months after HK gene delivery, the systolic blood pressure of experiment group was significantly decreased compared with the control group [(163±13) mm Hg vs (217±16) mm Hg, P<0.01](1 mm Hg＝0.133 kPa). Compared with sham rats, the rats in simple operation group and control group had much more renal interstitial collagen deposition and more serious fibrosis performance, but renal interstitial collagen deposition and fibrosis were significantly ameliorated in the rats of experiment group. In addition, the tubulointerstitial injury index of HK transgenic rats was significantly lower than that of the rats in control group (1.33±0.73 vs 3.01±0.62, P<0.01). Up-regulating expression of bradykinn B2 receptor protein and down-regulating expression of AT1 receptor and p-MAPK protein were found in renal tissues of experimental group after three months（P<0.05）. Conclusion HK gene delivery significantly alleviates renal interstitial fibrosis in rats with 5/6 nephrectomy through regulating the expression of bradykinin B2 receptor, AT1 receptor and p-MAPK in renal tissue.

Objective To investigate the role of RhoA-Rock signaling pathway in the process of rat peritoneal mesothelial cells (RPMCs) epithelial-mesenchymal transition (EMT) induced by transforming growth factor β1 (TGF-β1). Methods Primary RPMCs were cultured in vitro. After synchronization for 24 hours, RPMCs were randomly assigned to 4 groups: group A (control), group B (TGF-β1, 10 μg/L), group C (10 μg/L TGF-β1+10 μmol/L Y-27632, an inhibitor of Rock, pretreated for 2 hours with Y-27632 before TGF-β1 stimulation), group D(Y-27632 alone, 10 μmol/L). Growth arrested and synchronized RPMCs were stimulated by 10 μg/L TGF-β1 for different time. The mRNA and protein expression levels of E-cadherin, α-SMA and collagenⅠwere measured by RT-PCR and Western blotting respectively. The protein expression level of vimentin was measured by Western blotting. Active RhoA was extracted by Plasma Membrane Protein Extraction Kit, then it was assessed by Western blotting. Results (1) TGF-β1 stimulation elicited a robust increase in RhoA activity in time-dependent manner, which was (2.57±0.52) folds compared with control group (P<0.05) after 10 min stimulation. RhoA activity peaked at 1 hour, which was (4.35±0.41) folds compared with control group (P<0.05). (2) TGF-β1 up-regulated mRNA and/or protein expression of α-SMA, vimentin and collagenⅠ, and down-regulated mRNA and protein expression of E-cadherin in RPMCs. (3) The Rock inhibitor Y-27632 effectively revered TGF-β1-induced expression of α-SMA, collagenⅠand vimentin. The mRNA levels of α-SMA and collagenⅠdecreased by 53.8% and 55.7%, and the protein levels of α-SMA, vimentin and collagenⅠ decreased by 42.6%, 60.1% and 58.1% compared with TGF-β1-stimulated groups (P<0.05). But Y-27632 had no effect on the level of E-cadherin. Conclusions RhoA-Rock signaling pathway may mediate EMT induced by TGF-β1 in rat peritoneal mesothelial cells. RhoA-Rock pathway may be the potential therapeutic target in the progress of peritoneal fibrosis.

Objective To investigate the effect of fluvastatin on activation of nuclear factor kappa B（NF-κB）induced by angiotensin Ⅱ (AngⅡ) in rat kidney tubular epithelial cells (NRK-52E). Methods NRK-52E cells were divided into （1）control group；（2）AngⅡgroups with different concentration and time；（3）AngⅡ（10-6 mol/L）+SB203580 （10 μmol/L）group；（4）AngⅡ（10-6 mol/L） +different fluvastatin concentration （10-7, 10-6, 10-5 mol/L）groups；(5)AngⅡ（10-6 mol/L） +fluvastatin （10-5 mol/L） +mevalonate （10-4 mol/L）group. Electrophoretic mobility shift assays (EMSA) was used to detect NF-κB activation. Phosphorylation of cellular p38 mitogen-activated protein kinase (p38MAPK) was determined by Western blot. Monocyte chemoattractant protein (MCP)-1 mRNA was determined by RT-PCR. Results AngⅡ stimulated the DNA-binding activity of NF-κB，phosphorylation of p38MAPK and up-regulated the expression of MCP-1 mRNA in cultured NRK-52E cells in a dose-dependent manner(P<0.01). Ang Ⅱ(10-6 mol/L) induced a rapid (5 minutes) elevation of the p38MAPK phosphorylation. NF-κB DNA binding activity was increased at as early as 30 minutes(P<0.01), peaked at 2 hours after AngⅡ treatment(P<0.01). This stimulatory effect of AngⅡ on NF-κB was blocked by SB203580 (a specific inhibitor of p38MAPK) (P<0.01). Incubation of cells with fluvastatin significantly inhibited the AngⅡ-induced NF-κB activation and expression of MCP-1 mRNA in dose-dependent manner (P<0.05). Exogenous mevalonate (10-4 mol/L) prevented the effect of fluvastatin on NF-κB activation (P<0.05). Conclusions Fluvastatin reduces AngⅡ-induced NF-κB activation via the p38MAPK pathway in NRK-52E cells. Such effect of flurastatin is partly through blocked by mevalonate.

Objective To observe the changes in the expression of renal tissue TNF-α , NF-κB and the interrelation to renal cell apoptosis, and their influences of Inula Britannica(an inhibitor of inflammatory signal pathway) in exhausted swimming rats, and to investigate the role of inflammatory signal pathway. Methods Forty-eight male Wistar rats were randomly divided into three groups: control group(CN, n=8), exhaustive swimming group (ES, n=24) and Inula Britannica group(IB, n=16). The rats of CN were quiet without swimming. The rats of ES swam to exhaustion and were sacrificed at immediately (ESI, n=8), 6 hour (ES 6 h, n=8) and 24 hour (ES 24 h, n=8) after exhaustiing swimming. The rats of IB group took orally Inula Britannica at the dose of 25 ml/kg body weight at 24 h before swimming and then swam to exhaustive state. The rats of IB group were sacrificed at 6 hour (IB 6 h, n=8) and 24 hour (IB 24 h, n=8) after exhaustiing swimming. The renal cell apoptosis was measured by the method of terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL). The expression of TNF-α in renal tissue was examined by immunohistochemistry. The changes of NF-κB in renal tissue were measured by flow cytometry and immunohistochemistry. The interrelation between TNF-α and NF-κB was analyzed by Pearson method, and the interrelation between TNF-α, NF-κB and renal tissue cell apoptosis was analyzed by Spearman method. Results The number of renal tissue apoptotic cells was increased progressively from ESI to ES 24 h rats (P<0.05). Immunohistochemistry staining showed that the positive expressions of renal tissue TNF-α and NF-κB were increased progressively at 0 h (0.136±0.009, 0.129±0.011), 6 h (0.171±0.011, 0.166±0.009) and 24 h (0.229±0.008, 0.218±0.019) after exhaustiing swimming in ES compared with control group (0.109±0.010, 0.095±0.010) ( all P<0.05). The similar changes of renal tissue NF-κB was also revealved by flow cytometry. The expression of TNF-α was positively correldted with NF-κB （r=0.955, P<0.01）, and renal cell apoptosis was also positively correlated with TNF-α and NF-κB （r=0.953, r=0.939, P<0.01） in ES rats. Pretreatment with Inula Britannica, inhibited the up-regulation of expressions of renal tissue TNF-α（6 h：0.142±0.012, 24 h：0.130±0.010） and NF-κB（6 h：0.138±0.010, 24 h：0.136±0.011） induced by exhausting swimming. Conclusion Overtraining can induce the up-regulating expressions of renal tissue TNF-α and NF-κB, and Inula Britannica can partly counter the above changes in exhaustied swimming rats, which may be one important mechanisms of overtraining-induced renal tissue cell apoptosis and the anti- apoptosis effect of Inula Britannica.