Objective To investigate the possible injury mechanism of human kidney proximal tubular epithelial cell-2 (HK-2) induced by aristolochic acid (AA). Methods Cultured HK-2 cells were divided into 4 groups:normal control,treated by AA at the concentration of 30,60 and 120 ?mol/L for 48 h respectively. The morphological changes were observed by inverted phase contract microscopy. The cell viability was measured by the Cell Counting Kit-8 (CCK8) assay. Apoptotic cells were identified by flow cytometry. Expression of active Caspase-3 was measured by Western blot analysis. Automatic biochemical analyzer was used to detect the contents of LDH and ?-N-Acetylglucosaminidase (NAG) in the supernatant. The expression of E-cadherin and a-SMA was detected with laser scanning confocal microscope (LSCM). Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of TGF-?1 and collagen Ⅲ in the supernatant quantitatively. Results AA inhibited HK-2 cells proliferation,induced cell apoptosis and activated Caspase-3 expression,and increased the LDH and NAG levels. All of these were in a concentration-dependent manner. AA at the concentration of 60 ?mol/L inhibited E-cadherin expression,increased ?-SMA expression and TGF-?1 and collagen Ⅲ secretion. Conclusion AA inhibits cell proliferation,induces apoptosis and epithelial-mesenchymal transition (EMT) in HK-2 cells. AA at relatively low concentration (≤60 ?mol/L) mainly induces EMT in HK-2 cells,while,that at high concentration (≥120 ?mol/L) causes apoptosis and cytotoxicity.

Objective To compare the effect of hemodiafiltration(HDF) on the incidence rate of intradialytie hypotension. Methods We followed up eighteen patients with symptomatic hypotension history and divided them into group A and group B.Group A was treated with conventional hemodialysis(CHD) and HDF, Group B adopted HDF with low temperture hemodialysis. The number of hypotensive episodes, blood temperature during cardiopulmonary bypass, energy transfer rates were assessed. Results The incidence of symptomatic hypotension of patients was found lower in CHD (40%) than in HDF (6%),P<0.01. Energy transfer rates were founded higher in CHD than in HDF and low temperture hemodialysis, P<0.01. Conclusions HDF significantly reduced hemodialysis-related side effects and increased the tolerance ability of patients.

Objective To investigate the localization of Smad protein 2, 3, 6, 7 and their changes of expression in experimental interstitial fibrosis model in rats. Methods Thirty-six rats were divided into normal control, sham operation, and unilateral ureteral obstruction(UUO) groups, and were sacrificed at postoperative day 3, 7, 14, 21. The level of TGF-?1 mRNA was examined by RT-PCR. The sites and levels of expression of Smad protein 2, 3, 6, 7 were examined by immunohistochemistry staining and Western blot . Renal fibrosis was assessed by measuring tissue hydroxyproline. Results Compared to sham operation group, TGF-?1 mRNA was significantly increased in UUO rats, and this trend was positively correlated to increased hydroxyproline content. Immunohistochemistry staining studies indicated that Smad protein 2, 3 mainly expressed in renal tubular cells, rarely in glomeruli, and Smad protein 6, 7 were presented in both the glomeruli and proximal renal tubular cells. Expression of the Smad protein 2, 3 were significantly increased from day 3 to 21 after UUO, while the Smad protein 6, 7 were significantly reduced in the obstructive kidney. Conclusions TGF-?1/Smad signaling is involved in the progression of renal tubulointerstitial fibrosis. The absence of up-regulation of these anti-Smads proteins may be the major cause of the interstitial fibrosis in this model.

To evaluate the effects of interleukin-1 and interleukin-4 on the expression of p27 (one of the negative cell-cycle proteins) and CDK 2 (cell-cycle depending kinase, one of the positive cell-cycle proteins) on renal mesangial cells of rat in vitro, and to analyze their significance. The results showed that the proliferation of mesangial cells was induced markedly by interleukin-1, while the expression of p27 was down-regulated and that of CDK 2 was up-regulated. All of above changes were prevented by interleukin-4. The results suggested that the responses of rat renal mesangial cells to interleukin-1 and interleukin-4 were closely related to the changes in the expression of p27 and CDK 2.

Objective To investigate the localization of Smad2, Smad3, Smad4, and Smad7 proteins and their expression changes in the 5/6 subtotally nephrectomized rat kidney. Methods The rat model of chronic renal failure was established by performing 5/6 subtotally nephrectomy (SNx) and rats in the control group underwent sham-operation. The rats were sacrificed at 4, 8, and 12 week after operation. The sites and levels of expressions of Smad2, Smad3, Smad4, and Smad7 proteins were examined by immunohistochemical staining. Renal fibrosis was assessed by measuring tissue hydroxyproline. Results Immunohistochemical staining indicated that Smad2, Smad3, and Smad4 proteins were mainly expressed in glomeruli and renal tubular cells, while Smad protein 7 was expressed in glomeruli, but rarely in proximal renal tubular cells. Expressions of Smad2, Smad3, and Smad4 proteins in glomeruli were significantly increased during 4-12 weeks after 5/6 nephrectomy, but the expression level of Smad protein 7 was significantly decreased, but accompanied increase of hydroxyproline content in the renal tissues. Conclusion These results indicate that TGF-?/Smad signaling is involved in the progress of chronic glomerulosclerosis. The high level expressions of Smad2, Smad3, and Smad4 proteins and the down-regulation of Smad7 protein may be the major cause of the glomerulosclerosis in this model.

0.05).The incidence rate of diarrhea in IP group was 48.4% it was significantly higher than GP group(P

Objective To investigate the effect of over-expression of ROBO4 on permeability of human renal glomerular endothelial cells (HRGECs) in high glucose medium.Methods HRGECs infected with recombinant lentiviral vector ROBO4 were cultured in high glucose or low glucose medium in vitro.The protein levels of ROBO4 and ARF6 in each group were detected by Western blotting.The endothelial permeability was measured by the effiux of fluorescein isothiocyanate-dextran (FITC-Dextran)permeated through the monolayer endothelial cells using Transwell cell model system.The cell viability after lentivirus transfection was measured by CCK8 assay.Results The transfection rate of lentiviruses in HRGECs reached 80％ 72h after,and obvious overexpression of ROBO4 protein was in transformed cells compared with the empty vector group (P＜0.05).The lentivirus-mediated ROBO4 transfection did not affect cell viability of HRGECs.Compared with the low glucose group,the expression of ROBO4 increased obviously after 12h,but declined after 24h (P＜0.05),and reached to minimun after 72h (P＜0.05).On the contrary,the expression of ARF6 increased after 12h,and the increase reached to the maximum after 72h (P＜0.05).Furthermore,the vascular permeability increased gradually after 24h,and reached to the maximum after 72h (P＜0.05) in high glucose group.Compared with the empty vector group,the over-expression of ROBO4 inhibited the expression of ARF6 significantly,and the FITC-Dextran permeability reduced obviously.Conclusion Over-expression of ROBO4 may significantly enhance the barrier functions of HRGEC in high glucose medium,and ROBO4 activation may be a potential therapeutic approach in diabetic nephropathy.

AIM:To approach the therapeutic effects of mycophenolate mofetil(MMF)in hormonal resistance nephrotic syndromes.METHODS:Patients were diagnosed as nephrotic syndrome and treated with prednisone at the dose of 1 mg?kg-1?d-1 for over 8 weeks,and 24 patients with unsatisfactory results or were palindromic were selected,and several patients in the 24 patients had been treated with cyclophosphamide or cyclosporine A.All patients were treated with MMF combined with low dose hormone.The initial dose of MMF was 1.0-1.5 g/d for 3 months,later the dose were reduced,and the maintenance dose of MMF was 0.5-1.0 g/d,the dose of prednisone was 5-20 mg/d,the follow-up visit period more than six months.The changes on urine protein,serum albumin,liver function,renal function were compared before and after treatment.RESULTS:Before and after treatment,urine protein decreased from(3.4?1.7)g/d to(0.9?0.2)g/d,serum albumin increased from(19.6?5.4)g/L to(36.1?7.7)g/L.serum creatinine level decreased from(105.7?6.4)?mol/L to(90.1?5.8)g/L.20 patients(83.3%)pathogenetic condition were relieved,15 patients(65.2%)were with complete remission.5 patients(20.8%)were partially recovered,and 4 patients(16.6%)had no response.The adverse effects were observed,including gastrointestinal events(n=8,33.3%),bacterial pneumonia(n=4,16.6%),herpes zoster(n=1,4.1%),hepatic function mild damage(n=3,12.5%).CONCLUSION:It is safe and effective to combine MMF with low dose hormone in treatment of hormonal resistance nephrotic syndrome,which could become a therapeutic option for refractory nephrotic syndrome(RNS).

[Objective]To investigate the relationship of TCM syndrome patterns with gastrointestinal motility and hormones in functional dyspepsia (FD). [Methods] Among 30 cases of dyskinetic FD, 16 with liver depression and qi stagnation were allocated to Group B, 14 with spleen and stomach deficiency to Group C and 20 healthy volunteers served as control group ( Group A). Gastric evacuation, elctrogastrogram (EGG), serum motilin and migrating motor complex (MMC) wave of gastric antrum and duodenum during digestion were examined and analysed statistically. [Results]Gastric evacuation, contraction index and amplitude of phase II MMC wave, contraction amplitude and coordinated contraction of phase III MMC wave, and the main power of emptying EGG in empty stomach were decreased in Group B and Group C as compared with those in Group A,and those in Group C were lower than those in Group B, the differences being significant (P

Objective To examine the expression of BMP4 in CNS of the developing rat. Methods In situ hybridization histochemistry(ISHH) was carried out on tissue sections using specific digoxigenin\|labeled oligonucleotide probe. Results It showed that BMP4 mRNA positive cells were located mainly in cerebellum and olfactory at E16.Strong positive signal was seen in hypoglossal nucleus,and moderate signal also seen in spinocerebellar tract and spinal lemniscus at P1\|2.The number of BMP4 mRNA positive cells was increased in the frontal cortex,parietal cortex,and hippocampus subiculum at P1W.The peak of BMP4 expression was in cortex and periamydaloid cortex.Widely distributed BMP4 mRNA positive cells were detected in cortex and hippocampus of rats at P1M,strong positive signal was observed in temporal CNS at P3M,strong positive signal was observed in hippocampus,temporal corex and periamydaloid cortex,lateral nucleus of thalamus and paraventricular nucleus of hypothalamus.BMP4 mRNA positive cells were also found in corex,hippocampus,hypothalamus and thalamus at P18M.Conclusion\ These results indicated that BMP4 could play an important role in CNS development of rats.