Objective To investigate the characteristics and proper use of anticoagulants in hemodialysis (HD). Methods Thirty-one HD patients were enrolled in the study. Unfractionated heparins (UFH), dalteparin sodium or argatroban were used for HD anticoagulation respectively. Blood specimens were taken from the arterial line at the beginning (0 h) and at the end of HD (4 h), and from the arterial (2a) and the venous (2v) line respectively at 2 h of the HD session. Glass bead activated clotting time (gbACT), clot rate (CR) and platelet function (PF) were examined by Sonoclot analyzer. Prothrombin fragment 1+2 (PF1+2) and granule membrane protein-140 (GMP-140) were assayed by ELISA. Meanwhile, blood was taken from 8 healthy volunteers to examine the above parameters as control. Results (1) Compared with the control group, CR, PF1+2, PF, GMP-140 were increased significantly in all the patients (P<0.05). (2) UFH group: Compared with the 0 h point, gbACT of other time points increased significantly (P<0.05), CR, PF, and PF1+2 decreased significantly (P<0.05). Compared with the control group, gbACT increased (P<0.05) and CR decreased (P<0.05) significantly at the end of the sessions. (3) Dalteparin sodium group: Compared with the 0 h point, gbACT of 2a point increased significantly(P<0.05), CR and PF1+2 of 2a, 2v and 4 h points decreased significantly (P<0.05), meanwhile, the extents of increased gbACT and decreased CR from the arterial line were greater than those from the venous line. Compared with the control group, gbACT increased significantly at the end of HD session (P<0.05), but CR was not significantly different. (4) Argatroban group: There were no significant differences of gbACT between 0 h and other time points. CR of 2a, 2v points decreased obviously than that of 0 h point, and CR of 2v decreased more significantly. CR of 2a point was not different from the control group, while CR of 4 h point was greater as compared to control group. During the monitoring, PF1+2 tended to increase. Conclusions With intensive anticoagulant effect, UFH may induce the risk of hemorrhage not only during but also after the dialysis sessions. Dalteparin sodium, a good anticoagulant, is still related with the risk of hemorrhage during HD. Argatroban is an ideal anticoagulant for patients with the risk of hemorrhage.

Objective To observe the efficacy of calcitonin and bisphosphonates on renal osteopathy of maintenance hemodialysis （MHD）patients. Methods Forty-three MHD patients were randomly divided into two groups: A group and B group. All the patients were routinely received oral calcium carbonate 1.0 g tid and calcitriol 0.25 μg qd. Calcitonin (20U) hypodermic injection was given three times a week additionally during hemodialysis in A group. Patients in B group received bisphosphonates 70 mg once a week based on the therapy of A group. Serum levels of intact parathyroid hormone (iPTH), calcium, phosphorus, alkaline phosphatase (AKP), bone mass density (BMD) of lumbar spine and femoral neck, and the degree of bone ache (visual analogue scale, VAS) were assessed before the therapy and 3, 6 and 12 months after treatment. The adverse reactions were recorded during treatment. Results The levels of AKP and iPTH in both two groups decreased significantly after treatment. The above values of pre-treatment and 12 months after treatment were as follows： AKP（U/L）of A group 244.05±41.99 and 148.35±27.71，of B group 245.60±40.86 and 143.40±28.03；PTH（ng/L） of A group 697.5±119.7 and 267.4±45.9，of B group 708.2±120.3 and 277.6±41.9 （all P<0.05）. While the levels of calcium and phosphorus did not change obviously during treatment（P>0.05). BMD was not improved at 3, 6 mouths and became better at 12 mouths after treatment. As compared to pre-treatment, BMD of lumbar spine（g/cm2） in A group was 1.062±0.223 vs 1.202±0.251，in B group 1.033±0.152 vs 1.189±0.225; BMD of femoral neck（g/cm2）in A group was 0.993±0.108 vs 1.067±0.095，in B group 0.947±0.083 vs 1.018 ±0.217(all P＜0.05）. The scores of VAS also decreased significantly at 3, 6, 12 months after treatment（P<0.05）. No severe adverse reaction was found during the treatment. Conclusions Utilization of calcitonin and combination with bisphosphonates during hemodialysis can effectively preserve the BMD and prevent bone loss in MHD patients and is well tolerated. No significant difference of therapeutic effect is observed between using calcitonin or combination with bisphosphonates.

Objective To investigate the stroke occurrence of chronic kidney disease (CKD) and its related factors, especially the carotid atherosclerosis. Methods The data of stroke occurrence in 700 CKD patients hospitalized in Renji Hospital during 2007 were analyzed retrospectively. The incidences of stroke were compared among CKD Ⅰ-Ⅱ, CKDⅢ-Ⅴ non-dialysis patients and dialysis patients. Carotid atherosclerosis of 409 CKD patients was examined by color Doppler ultrasound. The related factors were selected by Spearman correlation analysis and Logistic regression analysis. Results Of 700 CKD patients, 67 cases (9.57%) experienced at least one episode of stroke, which was much higher than that of general population. The related factors of stroke in CKD included GFR, age, SBP, CRP, Lpa, serum glucose, pre-albumin, HDL and carotid atherosclerosis. Logistic regression revealed that SBP (β=1.021, P=0.042), CRP(β=1.008, P=0.024) and carotid atherosclerosis (β=3.456, P=0.025) were risk factors of stroke in CKD. Incidence of carotid atherosclerosis was high (50.37%) in CKD patients, besides it was significantly higher in CKD patients with stroke history as compared to those without stroke history (80.0% vs 47.4%, P<0.01). Conclusions The incidence of stroke is quite high in CKD patients, which is closely associated with hypertension, inflammation and glyeolipid metabolism disorder. Carotid atherosclerosis is common in CKD patients with stroke, which may be helpful in screening cerebrovascular diseases in CKD patients.

Objective To prospectively investigate the value of urinary neutrophil gelatinase-associated lipocalin (NGAL) in the diagnosis of acute kidney injury (AKI) following adult cardiac surgery. Methods Twenty-nine hospitalization patients undergone cardiac surgery were enrolled in the study. Serial blood and urinary samples were collected immediately before incision and at various time intervals after surgery. The primary outcome measure was acute kidney injury, defined as a 50% increase in the level of serum creatinine (Scr) from baseline. Results Fourteen of 29 developed acute kidney injury. The diagnosis time point with Scr was at 24 (10, 48) h after cardiac surgery. By contrast, the concentration of urinary NGAL rose from a median of 3.42(1.60, 9.92) μg/L at baseline to 20.51(13.42, 50.02) μg/L at admission to ICU (P=0.006), and the median concentration of urinary NGAL in patients who developed AKI was significantly higher at admission to ICU compared with patients who did not develop AKI [ 20.51 (13.42, 50.02) μg/L vs 2.91 (0.72,8.61) μg/L, P=0.002]. As to urinary NGAL at admission to ICU, the area under the receiver-operating characteristic (ROC) curve was 0.824, the sensitivity was 85.7% and the specificity was 80.0% with a cutoff value of 10.95 μg/L. Significant correlation was found between urinary NGAL at admission and the level of Scr at 24 h in ICU (r=0.545, P=0.002), as well as estimated GFR(r=-0.546，P=0.002). Conclusion Urinary NGAL concentration is significantly higher in patients developing postoperative AKI at the early time of admission to ICU, which may be a useful early biomarker of AKI after adult cardiac surgery.

Objective To explore the effect of Numb on tubular epithelial-to-mesenchymal transition(EMT) in rat proximal epithelial cells. Methods NRK52E cells were treated with different concentrations of recombinant human transforming growth factor-β1 (TGF-β1) (0, 1, 5, 10, 15, 20 μg/L) for 48 h or 10 μg/L TGF-β1 for different times (0, 24, 48, 72 h) in vitro. The expressions of E-cadherin, α-smooth muscle actin(α-SMA) and Numb in NRK 52E cells were detected by RT-PCR, Western blot and immunofluorescence staining. Meanwhile Numb siRNA oligo was transfected into NRK 52E cells with lipofectamine before TGF-β1 treatment, then Western blot was applied to detect the protein expression of E-cadherin, α-SMA and Numb in NRK52E cells. Results TGF-β1 could induce EMT in NRK52E cells in dose- and time-dependent manner. During the progress of TGF-β1-induced EMT, the protein expression of Numb in 5, 10, 15, 20 μg/L group was 1.33 folds (P=0.024), 1.39 folds (P=0.035), 1.45 folds (P=0.025), 1.51 folds (P=0.000) respectively as compared to 0 μg/L group. Likewise, the protein and mRNA expression of Numb in 24 h, 48 h, 72 h group was 1.48 folds (P=0.046) and 1.56 folds (P=0.012), 1.54 folds(P=0.011) and 1.82 folds (P=0.008), 1.79 folds (P=0.028) and 1.82 folds (P=0.002) respectively as compared to 0 h group. Moreover, large amount of Numb was accumulated in the cytoplasm. Down-regulation of Numb expression by siRNA transfection did not influence the basal expression of E-cadherin and α-SMA in NRK 52E cells, but attenuated the progression of EMT in NRK52E cells induced by TGF-β1. The up-regulation of α-SMA protein was reduced to 18.1% (P=0.004) while the down-regulation of E-cadherin protein was reversed to 2.19 folds (P=0.004). Conclusion Numb can promote EMT in rat proximal epithelial cells.

Objective To investigate whether the activation of secretory prophospholipase A2 (sPLA2) plays the role in the pathophysiological mechanism of acute aristolochic acid nephropathy(AAN) in rats. Methods Wistar rats were randomly divided into two groups. Model group received decocted Aristolochia Manshuriensis Kom 30 g&#8226;kg-1&#8226;d-1 by gavage for 7 days following tap water in same way for additional 7 days. Control group received only tap water by gavage at parallel time. The renal pathological changes were observed at the 4th, 8th and 14th day. The injury of renal tubules and interstitium was observed under light microscope following a semi-quantity grade. The level of Scr was measured to evaluate glomerular function. Urinary N-acetyl- beta-glucosaminidase (NAG) was tested as renal tubular injury marker. The activity of sPLA2 in serum was detected by manifesting the color of thiols in the substrate. The protein expression of renal cortex and medulla COX-2 was analyzed by Western blot. The metabolic products of protaglandins (PGs) including 6-keto-PGF1α and TXB2 in the plasma and urine were assayed by radioimmunoassay. The ratio of 6-keto-PGF1α/TXB2 was calculated. Results After Aristolochia administration, the tubulointerstitial injury and Scr increased in AA rats and reached the peak at the 8th day, the tubulointerstitial injury index(8.14±2.55 vs 1.50±0.71, P<0.05 ) and Scr[(0.24±0.10) vs (0.19±0.02) μmol/g, P<0.05] increased significantly in AA rats compared with control group. The activity of sPLA2 (μmol&#8226;min-1&#8226;mg-1) in AA group elevated by 1.3-fold compared to control group at 8th day (133.15±17.05 vs 101.3±16.07, P<0.05), while the expression of COX-2 in renal cortex increased (1.16±0.36 vs 0.69±0.28, P<0.05) with no change in renal medulla. Even though the levels of serum 6-keto-PGF1α and TXB2 did not change obviously in both AA and control group, but urinary levels of 6-keto-PGFlα and TXB2 increased by 2-fold and 3-fold in AA group compared to control group, respectively (all P<0.05), while the ratio of 6-keto-PGF1α/TXB2 decreased significantly (207.53±17.52 vs 296.64±51.31, P<0.05). All of above changes recovered to the control level at the 14th day except the tubulointerstitial injury index. Conclusion Serum sPLA2 is activated in the rats with acute kidney injury induced by aristolochic acid, which accompanied by up-regulated expression of COX-2 in renal cotex and increased the metabolic products of vasoconstrictive PGs in urine. These changes may participate the mechanism of renal peritubular ischemia in AAN.

Objective To examine whether tissue inhibitor of metalloproteinases-1 (TIMP-1) is involved in arterial calcification of chronic renal failure(CRF) rats. Methods CRF model was induced in male Wistar rats by gavage daily with 2% adenine 250 mg/kg. The calcification of aorta, femoral artery, renal artery and coronary artery was evaluated histomorphometrically by von Kossa-stained sections at 2-, 4-, 6- and 8-week respectively. RT-PCR and Western blot were used to observe the expressive levels of TIMP-1 mRNA and protein. Expressions of TIMP-1, osteopontin (OPN) and core binding factor α1(Cbfα-1) protein were analyzed by immunhistochemistry. Results Serum urea nitrogen, creatinine, inorganic phosphate, calcium-phosphorus product and intact parathyroid hormone (iPTH) increased significantly in the model animals compared with control group after 2 weeks(P<0.01). Medial calcification was found in above four arteries of model groups after 6 weeks. RT-PCR and Western blot showed that TIMP-1 expression of model group was significantly higher than that of control group (P< 0.05), and obviously elevated in a time-dependent manner. The expression of TIMP-1 and OPN in calcified aortic smooth muscle cells increased obviously (P<0.05), and positive immunostaining of Cbfα-1 was found. The expression of TIMP-1 was positively correlated with OPN and Cbfα-1（r=0.317, P=0.000; r=0.485, P=0.000）. Conclusions The pathology of arterial calcification in CRF rats induced by adenine is similar to CRF patients, which may serve as a useful model of CRF with arterial calcification. The up-regulation of TIMP-1 seems to participate in the formation and development of vascular calcification in CRF.

Objective To investigate the effect of syndecan-4 on the proliferation and extracellular matrix (ECM) secretion of human mesangial cells(HMC) stimulated by basic fibroblast growth factor (bFGF) and to evaluate the role of syndecan-4-PKCα pathway. Methods The expression of syndecan-4 in HMC was observed by immunofluorescence. After the down-regulation of syndecan-4 in HMC by RNA interference, the cell proliferation was detected by MTT. The secretion of fibronectin(FN), type Ⅳ collagen, typeⅠcollagen was assessed by ELISA. The copy number of syndecan-4 and PKCα was measured by fluorescent quantitation PCR at different time points. Results Syndecan-4 was expressed in HMC. bFGF could promote the cell proliferation and ECM secretion together with the PKCα copy number per million house-keeping genes of HMC, which could be reversed by the syndecan-4 siRNA transfection (MTT: 48-60 h, P<0.01; FN: 24 h, P<0.01, 48-96 h, P<0.05; type Ⅳ collagen: 72-96 h, P<0.05; PKCα: 0 h, P<0.05, 12-48 h, P<0.01）. Conclusion Syndecan-4 may regulate the proliferation and ECM secretion of HMC stimulated by bFGF through syndecan-4-PKCα pathway.

Objective To investigate the effect of 12-lipoxygenase(12-LO) on the p27kip1 expression in diabetic glomeruli. Methods Mesangial cells were exposed to 12-LO product 12(S)-HETE (10-7 mmol/L) with or without p38 MAPK (p38) inhibitor (SB203580, 1 μmol/L) for 24 hours. Rats fed with high fat diet received low dose streptozotocin (STZ, 35 mg/kg, IP injection) to develop type 2 diabetes and were divided into 2 groups: low dose STZ, low dose STZ+12-LO inhibitor cinnamyl-3,4-dihydroxy-α-cynanocinnamate (CDC, 8 mg/kg) treatment. Rats fed with regular chow were divided into two groups: controls, CDC treatment. The rats received injection of CDC or vehicle subcutaneously in the hind leg. CDC or vehicle injection was performed three times weekly on alternate days. All the rats were sacrificed after 4 weeks. Wild type and 12-LO knockout C57BL/6 mice were divided into 4 groups: wild type control, 12-LO knockout, STZ- induced wild type type 1 diabetes and STZ-induced 12-LO knockout type 1 diabetes. All the mice were sacrificed after 16 weeks. Urine, blood, kidney cortical tissue and isolated glomeruli by sieving method were collected at the end of study respectively. Western blot and immunohistochemistry for target protein were performed respectively. Results Inhibition of p38 activation could significantly reduce p27kip1 expression induced by 12(S)-HETE in mesangial cells (P<0.01). Increased glomerular volume, microalbuminuria, elevated glomeluli p38 activation, p27kip1 expresssion in type 2 diabetic glomeruli was decreased after CDC treatment (P<0.01). Compared with wild type diabetic mice, glomerular p38 activation, p27kip1 expresssion and extracellular matrix accumulation in the 12-LO knockout diabetic mice were significantly decreased（P<0.01, respectively）. Conclusions 12-LO induces p27kip1 expression via p38 pathway in diabetic glomeruli.

Objective To investigate the expressions of intermedin /adrenomedullin 2 (IMD/AM2) and its receptor calcitonin receptor-like receptor(CRLR) in the kidney of rats after renal ischemia reperfusion injury(IRI). Methods Male Wistar rats were divided randomly into two groups: sham group and operation group. Renal IRI model was induced by clamping both renal arteries. Blood and kidney were harversted at 0 h, 6 h, 12 h, 24 h, 48 h, 72 h after reperfusion, respectively. Renal histological changes were semi-quantitated. Expressions of IMD and CRLR in the kidney were detected by Western blot, and the content of IMD in serum was measured by radioimmunity at 12 h, 48 h, 72 h after reperfusion. Results Kidneys of renal IRI model rats displayed significant pathologic changes, and the changes were much severer at 48 h after reperfusion. The expressions of IMD and CRLR in kidney were significantly up-regulated at 12 h, 48 h, 72 h after renal IRI (P<0.01). The level of IMD in serum increased at 12 h, 48 h, 72 h after renal IRI(P<0.05). Conclusion The expressions of IMD and its receptor are up-regulated in the kidney after renal IRI, which may participate in the pathophysiological changes induced by renal IRI.