Aim To investigate the effects of fluvastatin on the cellular proliferation and the expression of connective tissue growth factor(CTGF)and type Ⅳ collagen(Col Ⅳ)in rat mesangial cells(MCs)induced by transforming growth factor-?1(TGF-?1).Methods MCs are divided into the following groups according to different factors:control group,TGF-?1 group,TGF-?1 plus fluvastatin(Flu,different concentrations)groups.The influence of Flu on rat proliferation of MCs was detected by MTT.The expression of CTGF was measured with RT-PCR and Western blot respectively.The secretion of Col Ⅳ protein was quantitated by ELISA.Results 5 ?g?L-1 TGF-?1 could stimulate proliferation of MCs and the expression of CTGF.Col Ⅳ in MCs significantly.Flu could inhibit proliferation of MCs and the expression of CTGF,Col Ⅳ in MCs induced by TGF-?1 in a dose-dependent manner:1 ?mol?L-1 Flu could suppress proliferation of MCs and downregulate the expression of CTGF,Col Ⅳ significantly,while 10 ?mol?L-1 Flu could just suppress the expression of CTGF protein significantly.Simultaneously exogenous CTGF could promote the expression of Col Ⅳ in a dose-dependent manner:2.5 ?g?L-1 CTGF could just upregulate the expression of Col Ⅳ.Conclusion Fluvastatin could inhibit proliferation of MCs and expression of CTGF-mediated Col Ⅳ in MCs.

Objective To explore the risk factors of bone density disorder and vascular calcification in non-dialysis chronic kidney disease (CKD) patients.Methods Clinical data of nondialysis CKD patients who were admitted to the First Affiliated Hospital of Fujian Medical University between January 2013 and June 2014 were retrospectively analyzed.Using dual energy X-ray absorptiometry to evaluate their bone mineral density (BMD) and T value.Patients were divided into normal BMD group (T≥-1),osteopenia group (-2.5 ＜ T ＜-1) and osteoporosis group (T≤-2.5).The vascular calcification was evaluated by pectoral computed tomography.Multi-factor stepwise logistic regression analysis was used to assess the risk factors for low bone density and vascular calcification in non-dialysis CKD patients.Results A total of 337 non-dialysis CKD patients were enrolled.There were 110 (32.6％) patients with normal BMD,and 146(43.3％) patients with osteopenia,and 81(24.0％) patients with osteoporosis.Gender,history of hypertension,25-hydroxy vitamin D and N-terminal osteocalcin shown statistical differences among three groups (all P ＜ 0.05).The incidence rate of 25-hydroxy vitamin D deficiency shown statistical difference among three groups (P=0.012).Further,the rates were increased with the decreased bone mass (x2=7.100,P=0.008).The other mineral bone disorders,such as hypocalcemia,hyperphosphatemia,low intact parathyroid hormone (iPTH) and high iPTH had no statistical difference among three groups (all P ＞ 0.05).Multi-factor stepwise logistic regression analysis revealed that increased iPTH (OR=1.938),and low bone density (OR=1.724) were independent risk factors for CKD patients with vascular calcification (all P ＜ 0.05),while women (OR=3.312) and vascular calcification (OR=1.742) were independent risk factors for CKD patients with low bone density (all P ＜ 0.05).Conclusion Increased iPTH and low bone density are independent risk factors for non-dialysis CKD patients with vascular calcification,while women and vascular calcification are independent risk factors for non-dialysis CKD patients with low bone density.

Objective To investigate the effects of erythropoietin(EPO)on the function of glomerular endothelial cells in rats with chronic renal failure(CRF). Methods The CRF model was established by a two stage 5/6 nephrectomy procedure in rats.Experimental rats were randomly divided into four groups:sham operation group (control group),CRF group,CRF rats treated with 30 U/kg EPO(low-dosage group)and with 50 U/kg EPO (high-dosage group).CRF rats received EPO by hypodermic injection for 6 weeks and then were sacrificed.Serum creatinine(Scr),blood urea nitrogen fBUN),urine protein,haematoglobin (Hb) and blood pressure were measured.The renal morphologie changes were evaluated on periodic acid-schiff (PAS) stained sections.The CD34 and CD31 expressions in glomerulus were detected by immunohistochemistry method.The mRNA of endothelin 1(ET-1),endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were detected by RT-PCR. Results The expressions of CD34 and CD31 protein in glomerulus,and the expressions of eNOS and VEGF roRNA in renal tissue were higher in EPO treatment group than those in CRF model group(all P<0,05).The expression of ET-1 mRNA in renal tissue was lower in EPO treatment group than that in CRF model group.In addition,the Scr,BUN,urine protein and blood pressure in EPO treatment group were significantly lower than those in CRF model group (all P<0.05).Haematoglobin in EPO treatment group was higher than that in CRF model group (P<0.05).Reanl pathological injury wss improved by EPO treatment in dose-dependent manner. Conclusion EPO can ameliorate renal pathological injury and renal function in rats with chronic renal failure,maybe through promoting the renovation of glomerular capillary endothelium and improving the function of glomerular endothelial cells.

Objective To investigate the effects of erythropoietin (EPO) on the number and function of peripheral blood endothelial progenitor cells (EPCs) from rats with chronic renal failure (CRF). Methods The model of chronic renal failure was established by a two-stage 5/6nephrectomy procedure in rats. Experimental rats were randomly divided into four groups (n =7,respectively): sham operation group, CRF group, CRF rats treated with 30 U/kg EPO (low-dosage group) and with 50 U/kg EPO (high-dosage group). CRF rats were given EPO by hypodermic injection for 6 weeks, then EPCs were isolated by density gradient centrifugation from peripheral blood mononuclear cells. The ability of cell proliferation, adhesion and vasculogenesis in vitro was further observed. Results Compared to sham operation group, the ability of cell proliferation,adhesion and vasculogenesis in vitro in CRF rats was remarkably decreased (P＜0.05, respectively).Such ability was promoted significantly in dose-dependent manner by EPO treatment (P＜0.05,respectively). Conclusion EPO can improve the number and ability of endothelial progenitor cells from rats with chronic renal failure.

Objectives To investigate the risk factors of acute renal injury (acute kidney injury) in patients with acute left heart failure.Methods Clinical data of 188 patients with acute left heart failure who were admitted to our hospital were retrospectively analyzed.Logistic regression analysis was used to assess the risk factors for AKI.Results Among 188 patients with acute left heart failure,incidence of acute kidney injury was 33.51％.Univariate and Multivariable logistic regression analyses showed that the independent predictors of acute kidney injury were lower baseline eGFR (OR=4.294,P ＜ 0.001) and anemia (OR=3.573,P=0.006).Conclusions The incidence of acute left heart failure complicated with AKI was high.Basic state of renal function and anemia were the independent risk factors for AKI.

Objective To observe the effect of fasudil on cytoskeleton reconstruction in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),as well as to study the protective mechanism of fasudil in the pathological changes of podocytes.Methods Conditionally immortalized mouse podocytes were treated with Ang Ⅱ (10-7 mol/L).The podocytes were pre-incubated for 30 min or 60 min with various concentrations of fasudil (10-8,10-7,10-5 mol/L),then Ang Ⅱ (10-7 mol/L) were added and furtherincubated for 24 hours.The cytoskeleton distribution of podocyte as indicated by F-actin and synaptopodin was observed by fluorescence microscopy and Western blotting.At the same time,the activity of Rho signal pathway that mediates actin filament polymerization was analyzed by measuring the extent of Rho-associated coiled-coil-containing protein kinase 1 (ROCK-1) and myosin phosphatase-1 (MYPT1).Results Compared to the control group,AngⅡ disrupted the podocyte actin cytoskeleton and significantly decreased the expression of synaptopodin (P ＜ 0.05),while fasudil stabilized the actin filaments,and improved the synaptopodin expression (P ＜ 0.05).The expression enhancement of ROCK-1 and MYPT1 by Ang Ⅱ were reduced significantly by fasudil (P＜0.05).Conclusion The cytoskeleton reconstruction of podocytes can be induced by Ang Ⅱ and inhibited by fasudil,which suggests that the protective effect of fasudil may be partially contributed to the Rho/ROCK signaling pathway.

Aim To investigate the effects of bone morphogenic protein(BMP)-7 on the expression of extracelluar matrix components(ColⅠand FN)in human renal proximal tubular cells(HK-2)induced by transforming growth factor-?1(TGF-?1),and to explore the inhibition of BMP-7 on renal interstitial fibrosis.Methods HK-2 cells were treated with TGF-?1,BMP-7,or a combination of both for 48 h.The expression of FN was assessed by indirect immunofluorescence.RT-PCR and Western blot were used to determine the mRNA and protein expression of FN and ColⅠ?1.Results Indirect immunofluorescence staining showed that FN staining in 3 ?g?L-1 TGF-?1 group was considerably stronger than that in control group.By an addition of 200 ?g?L-1 BMP-7,the expression of FN was significantly inhibited.RT-PCR and Western blot showed the mRNA and protein expression of FN and ColⅠ?1 were up-regulated by TGF-?1 after 48 hours significantly(P

Aim To investigate the effects of an angiotensin Ⅱ receptor blocker,candesartan, on aorta oxidative stress-LOX-1 pathway in salt-loaded stroke-prone spontaneously hypertensive rats (SHRSP).Methods 12-week-old salt-loaded SHRSP were treated with candesartan(1.0 mg?kg-1?d-1)or a diuretic, trichlormethiazide(TCM,1.6 mg?kg-1?d-1) or no treatment(n=6) in each for 2 weeks. Age-matched salt-loaded WKY rats were served as control(n=6).Systolic blood pressure(SBP)was measured weekly throughout the 2-week period by means of the tail-cuff method.Thoracic aortas were extracted and 24 h urine was collected.NAD(P)H oxidase subunits(p22 phox, p47 phox and gp91 phox)mRNA expression in aorta were assayed by real-time PCR. LOX-1 and type Ⅳ collogen mRNA expression were examined by RT-PCR. gp91 phox and LOX-1 protein expression in aorta were assayed by immunohistochemistry.Urinary albumin excretion was examined by ELISA.Results At the end of the 2nd week, SBP was significantly higher in salt-loaded SHRSP than that in salt-loaded WKY rats. Treatment with candesartan and TCM significantly decreased SBP in salt-loaded SHRSP at similar levels.NAD(P)H oxidase subunits (p47 phox and gp91 phox)and LOX-1 mRNA expression in aorta were markedly higher in salt-loaded SHRSP than those in salt-loaded WKY rats.Candesartan and TCM had the effect of reducing the systolic blood pressure at similar levels. Candesartan significantly down-regulated aorta p22 phox, gp91 phox,LOX-1 and type Ⅳ collogen mRNA expression and decreased urine albumin excretion in salt-loaded SHRSP(P

Objective To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to renal tubular epithelial-like cells under different conditions. Methods MSCs were obtained from rat marrow. MSCs were isolated by gradient density centrifugation and plastic adherence and then purified. Surface markers were identified with flow cytometry after amplification in vitro. The purified MSCs of the third passage were cultured respectively as follows: (1) control group: DMEM medium with fetal bovine serum(FBS). (2) all-trans retinoic acid (ATRA) group: DMEM medium with FBS, ATRA and ischemic reperfusion-injured kidney tissue homogenate. (3)combination group: DMEM medium with FBS, ATRA, ischemic reperfusion-injured kidney tissue homogenate, epidermal growth factor (EGF) and bone morphogenetic protein 7 (BMP-7). After 7 days, the MSCs were collected for alkaline phosphatase (AKP) staining, cytokeratin-18 and E-cadherin immunocytochemical analysis. Results The positive rates of the third passage MSCs in CD44, CD90 and CD29 were 97.8%±0.9%, 96.8%±1.4% and 97.6%±2.4%,respectively, but in CD11b/c and CD34 were only 13.2%±0.6% and 1.2%±0.5%. The MSCs in control group were spindle. The MSCs in ATRA group were round and elliptic. The MSCs in combination group became cobblestone-like cells after 7 days. AKP staining showed that tubular epithelial-like cells from MSCs in control group were negative, some above cells in ATRA group were positive and number of above cells increased in combination group. Compared with negative control group, the ratios of cytokeratin-18 positive cells in ATRA group and combination group were respectively increassed by 29.47%±1.08% and 47.52%±2.13% (all P<0.05), the ratios of E-cadherin positive cells in ATRA group and combination group were respectively increased by 14.88%±2.46% and 36.15%±1.13% (all P<0.05). Conclusion MSCs may differentiate by renal tubular epithelial-like cells under the induction of ischemic reperfusion-injured kidney tissue homogenate and ATRA in vitro, which are further differentiated under the combined induction of EGF and BMP-7.

Objective To clone and express the outer surface protein C ( OspC) from a Chinese Borrelia afzelli FP1 strain and to evaluate the immune protectivity of the recombinant OspC protein ( rOspC) . Methods The gene encoding OspC protein of Borrelia afzelli FP1 strain was amplified by polymerase chain reaction (PCR) and then inserted into pET-30a plasmid to construct the recombinant expression plasmid pET-30a-OspC. The transformed E. coli BL21 strains carrying pET-30a-OspC plasmid were induced by IPTG to express OspC protein. The expressed proteins were purified by Ni-IDA resin chromatography and analyzed by SDS-PAGE and Western blot assay. Indirect immunofluorescence assay ( IFA) was performed to detect anti-rOspC protein antibodies in serum samples from rabbits immunized with rOspC protein. In vitro neutral-ization test was performed for evaluation the immune protectivity of rOspC protein. Results The recombi-nant expression plasmid pET-30a-OspC was successfully constructed and highly expressed in E. coli BL21. A strong antigen-antibody reaction between the rOspC protein and polyclonal antibody against Borrelia afzelli FP1 strain was detected by Western blot assay. The titers of IgG in serum samples from rabbits immunized with rOspC protein were significantly elevated. The in vitro neutralization test indicated that 106/ml of Borre-lia afzelli FP1 strains were neutralized by every anti-OspC protein serum sample from the experiment group. Conclusion The rOspC protein showed a strong immune protectivity against Borrelia afzelli, which could be used in the development of polyvalent subunit vaccine against lyme disease.