Objective To explore the pathological changes of the middle-small arteries and veins of uremic patients in early stage.Methods Forty patients with chronic kidney disease at Ⅴ stage were randomly subjected in our study.Radial artery and cephalic vein about 0.5 to 1 cm in length were obtained by arteriovenous fistula operations.Cephalic veins from 4 trauma patients were taken as control group.Immunohistochemisty with anti-?-SMA(smooth muscle actin-alpha),BMP-2(bone morphology protein-2) and BMP-7(bone morphology protein-7) antibodies,alizarin Bordeaux staining,HE staining and Masson trichrome staining were carried out to investigate calcification and fibrosis.Results Compared with control group,the expressions of ?-SMA and BMP-2 were up-regulated obviously.Masson trichrome staining indicated fibrosis in the vein from uremic patients.Conclusion Fibrosis may be one of the early pathologic changes in the vein from uremic patients,suggesting the venous alterations may be related to cardiovascular disease of uremic patients.

Objective: To examine the effect of Benapril on atherogenesis and the plaque rupture in rabbits.Methods: Thirty four healthy male New Zealand white rabbits were randomly divided into 4 groups.Group C (control group) was fed normal diet for 10 weeks,group HL fed 1% cholesterol diet,group B fed 1% cholesterol diet and Benapril 5 mg/d.Ten rabbits of plaque rupture group were fed 1% cholesterol diet for 10 weeks with atherosclerosis induced by left carotid iliac damage.Ten weeks after the initiation of the diet,an angioplasty was performed.After angioplasty,the surviving rabbits( n =10) were randomized to receive benapril(5 mg/d,each) supplementation in drinking(B group, n =4) or no treatment(untreated group, n =4). The levels of CEC were measured by morphometrical counts at different periods(the 5th week,the 10th week).Before and 5,10 weeks after experiment,fasting blood samples were collected for serum total cholesterol(TC),high density lipoprotein cholesterol(HDL C),low density lipoprotein cholesterol(LDL C),and triglyceride(TG) assay.The levels of plasma circulating endothelial cells(CEC) were measured by morphometrical counts and the levels of plasma von Willebrand factor(vWF) were determined by ELISA.After sacrificing the separated, entire aortas were stained with oil red O and then processed for histological examination;planimetry was done with a computer system.Endothelial cells were confined by the presence of Factor Ⅷ related antigen as the specific cell marker.Results: The serum TC,TG,LDL C increased progressively in group B and group HL,but the levels of vWF in group HL were significantly higher than that in group B on the 5th and 10th respetively( P

The aim of this study was to explore the myocardial ultrastructure of experimental uremic rats and the effect of myocardiocyte succinate dehydrogenase(SDH) and free cytosolic calcium changes on membrane channel. By making a uremia model in rats, changes of SDH activation was observed by quantitative enzyme cytochemistry methods and membrane calcium channel was checked by laser scanning confocal microscopy. The results showed that with the uremia becoming heavier, myocardiocyte membrane was rolled and broken, myofibrils were swollen and broken. The quantity of mitochondria with SDH products significantly decreased, while activation of cell membrane calcium channel markedly increased and cytosolic calcium piled up. It is suggested that impaired myocardial membrane, decreased contents of mitochondrial function enzyme and cytosolic calcium overload are the pathophisiological basis leading to cardiac dysfunction in uremia.

To examine the effect of benapril on atherogenesis, circulating endothelial cells (CEC), and factor vWF. Twenty four healthy male New Zealand white rabbits were randomly divided into 3 groups. In group C (control group) the rabbits were fed normal rabbit diet for ten weeks, in group AS 1% cholesterol diet was fed, and in group B both 1% cholesterol diet and benapril 5mg/day were fed. Fasting blood samples were collected for the determination of serum total cholesterol (TC), high density lipoprotein cholesterol (HDL c), low density lipoprotein cholesterol (LDL c), and triglyceride (TG) on day 0, 5 and 10 weeks after feeding. The levels of plasma CEC were measured by morphometrical count and the levels of plasma vWF were determined by ELISA. After the animals were sacrificed, aortas were obtained, and they were stained with oil red O. The tissue was sectioned for histological examination. Planimetry was performed with a computer system. Endothelial cells were identified by the presence of factor VIII related antigen, which was the specific marker for endothelial cells. The results showed that the levels of serum Tc, TG, and LDL c were increased progressively in group B and group AS, but the levels of vWF in group AS were significantly higher than that in group B at the 5 th and 10 th weeks respectively ( P

Objective To investigate the therapeutic effect of endothelial progenitor cells(EPCs) in ameliorating rat progressive focal segmental glomerular sclerosis(FSGS) model induced by adriamycin.Methods Bone marrow mononuclear cells from male SD rats,after cultured by adherence method,were identified as EPCs.Female SD rats were divided into normal control group,adriamycin induced renal disease(ADR) group,EPCs transplantation group.ADR group and EPCs group underwent unilateral nephrectomy and received 5,3 mg/kg of adriamycin via tail vein 1 week and 2 weeks after operation,while the control group underwent sham operation and received 0.9% sodium chloride solution of equal volume.The whole body irradiation by 5 Gy X ray was done 1 week after the 2nd injection of adriamycin,then immediately 1?106 EPCs were transplanted via tail vein.The rats in control group and ADR group were only injected with 0.9% sodium chloride solution after whole body irradiation.The body weight and urine protein were measured before operation(0 week) and 4(1 week after EPCs transplantation),8,12 and 16 weeks after nephrectomy.Y chromatosome incorporation was detected with in situ hybridization at the 4th and 16th week.The histological and ultrastructural changes of kidney were evaluated at the 16th week.Results At the 4th and 16th weeks,Y chromatosome positive cells could be found incorporation in the area of glomerular and tubular epithelial cells.Since the 4th week,the weight of rats in both ADR group and EPC group became significantly less than that in control group and since the 8th week that in ADR group became less than that in EPC group(P

Objective To approach the selection of appropriate period and mode of blood purification for treatment of patients with severe acute pancreatitis(SAP)based on acute physiology and chronic health evaluationⅡ(APACHEⅡ)score. Methods The clinical data of 89 patients with SAP were retrospectively analyzed. They were assigned into two groups:the hemoperfusion(HP)and short continuous veno-venous hemodiafiltration(SCVVHDF) group(HP+SCVVHDF,49 cases)and the HP and hemodiafiltration(HDF)group(HP+HDF,40 cases). All the patients were evaluated by APACHEⅡscore. In the HP+HDF group,26 cases with APACHEⅡ<15 were in group A, while 14 cases with 15-25 in group B. In the HP+SCVVHDF group,31 cases with the score 15-25 were in group C, and 18 cases with<15 in group D. One week after the treatment,APACHEⅡscore,oxygenation index(PaO2/FiO2), C-reactive protein(CRP),serum creatinine(SCr),alanine transaminase(ALT)and mortality were observed in all the groups. Results Compared to those before treatment,the APACHEⅡ scores(group A:7.35±2.12 vs. 10.52±3.26,group B:14.35±4.76 vs. 18.43±4.08,group C:11.83±3.85 vs. 19.39±3.64,group D:6.92±2.54 vs. 11.61±2.19),the levels of CRP(mg/L,group A:85.28±23.48 vs. 195.23±56.77,group B:172.67±36.69 vs. 232.65±62.86,group C:112.43±29.48 vs. 257.29±68.39,group D:76.23±29.05 vs. 206.37±65.49),SCr (μmol/L,group A:107.56±73.01 vs. 225.81±119.06,group B:291.49±123.27 vs. 391.76±273.48,group C:254.89±104.37 vs. 403.62±261.53,group D:112.36±55.36 vs. 258.74±128.25)and ALT(U/L,group A:86.93±27.04 vs. 127.56±84.35, group B:116.34±43.98 vs. 189.15±102.85, group C:94.85±74.42 vs. 178.73±87.21,group D:88.49±29.32 vs. 138.24±90.58)after treatment were all decreased markedly in the four groups. The levels of PaO2/FiO2 were obviously higher in the 4 groups after treatment〔mmHg(1 mmHg=0.133 kPa), group A:293.42±31.26 vs. 253.60±26.62,group B:254.12±35.73 vs. 137.56±23.48,group C:283.21±37.48 vs. 131.96±0.45,group D:305.75±29.66 vs. 267.74±31.42〕,but no statistical significant differences in above indexes between group A and group D were found(all P>0.05),while the APACHEⅡ score,PaO2/FiO2,CRP, SCr and ALT were improved more significantly in group C than those in group B(P<0.05 or P<0.01). The mortality rate of those SAP patients with APACHEⅡscore<15 was lower than those in cases with APACHEⅡscore 15-20〔6.82%(3/44)vs. 24.44%(11/45),P<0.05〕. Conclusions Blood purification is an effective measure to save patients with SAP. The APACHEⅡ score used to select the mode of blood purification in appropriate period for treatment of SAP has guiding significance. Currently the modes of blood purification have limited value and cannot cure all SAP patients.

Objective To investigate the effect of hairy and enhancer of split 5(HES5)on transdifferentiation and apoptosis of renal tubular epithelial cells induced by TGF-β1 and its potential mechanism.Methods The differentially expressed genes in GSE66494 data were analyzed and screened.The mouse model of unilateral uretera obstruction(UUO)was established,and the expression level of HES5 was detected in the renal tissue.HK-2 cells were treated with 10 ng/mL TGF-β1 for 24 h to establish a tubular epithelial-mesenchymal transition(EMT)model,and then qRT-PCR and Western blotting were performed to detect the expression of HES5 at mRNA and protein levels.After HK-2 cells were transfected with the plasmid overexpressing HES5,the protein levels of fibronectin,collagen Ⅰ,vimentin,apoptosis markers Bax and Bcl2 were detected in 24 h later.Then,HK-2 cells were divided into Control group,siHES5 group,TGF-β 1 group,and siHES5+TGF-β1 group.The protein level of fibrosis and apoptosis markers were measured in above groups with Western blotting.TUNEL staining and flow cytometry were employed to detect cell apoptosis.Western blotting was applied to determine the protein levels of AKT,p-AKT,PI3K and p-PI3K.HK-2 cells overexpressing HES5 were treated with PI3K inhibitor LY294002,and the expression of vimentin was detected.Results The expression of HES5 was significantly up-regulated in both chronic kidney disease(CKD)and fibrotic kidneys of mice.Overexpression of HES5 promoted the synthesis of fibronectin,collagen Ⅰ,vimentin and Bax in HK-2 cells,and inhibited the expression of Bcl2(P＜0.05).HES5 knockdown not only down-regulated the expression of fibrosis markers,but also inhibited the apoptosis of HK-2 cells.Furthermore,HES5 knockdown inhibited the activation of PI3K/AKT signaling pathway induced by TGF-β1 in HK-2 cells(P＜0.05).Inhibitors of the PI3K/AKT signaling pathway inhibitor attenuated the induction of HES5 on vimentin.Conclusion HES5 knockdown inhibits the transdifferentiation and apoptosis in TGF-β1-induced renal tubular epithelial cells,which may be related to the decreased activity of the PI3K/AKT signaling pathway.

Objective To investigate the role of fibrinogen-like protein 2(Fgl2)in macrophage polarization during cisplatin(Cis)-induced acute kidney injury(Cis-AKI).Methods Twelve male wild-type(Fgl2+/+)mice and 12 Fgl2 gene knockout(Fgl2)mice,aged 8～10 weeks and weighing 20～25 g,were selected,and then after being administered with a single intraperitoneal injection of either saline or Cis for 3 d,they were randomly divided into 4 groups(n=6):Fgl2+/+Saline group,Fgl2+/+Cis group,Fgl2 Saline group and Fgl2 Cis group.Kidney function indicators such as serum creatinine(Scr)and blood urea nitrogen(BUN)levels were measured 3 d later,and kidney injury was assessed by HE staining.Western blot analysis was performed to evaluate the expression of Fgl2 and kidney injury molecule 1(Kim-1)in the renal tissues.RT-qPCR was conducted to assess the expression levels of Fgl2,Kim-1,neutrophil gelatinase-associated lipocalin(NGAL),IL-6,IL-12p40,IL-1β,inducible nitric oxide synthase(iNOS)and TNF-α in the renal tissues.Immunohistochemical assay was employed to detect the expression of Fgl2 and macrophages(F4/80+)in the kidneys.Immunofluorescence staining was utilized to examine the expression of macrophages(F4/80+)and M1-type macrophages(F4/80+CD86+)in the renal tissues.Flow cytometry was employed to analyze the expression of macrophages(F4/80+)as well as M1-type macrophages(F4/80+MHC Ⅱ+)and M2-type macrophages(F4/80+CD206+)in the renal tissues.Results Compared with the Fgl2+/+Saline group,the Fgl2+/+Cis group exhibited a significant decline in renal function(P＜0.05),a notable increase in pathological score of renal tubular injury(P＜0.05),and an obvious upregulation of renal tissue Fgl2 expression(P＜0.05).Compared with the Fgl2+/+Cis group,the Fgl2 Cis group demonstrated a significant decline in renal function(P＜0.05),an elevation in the expression of renal injury-associated molecules Kim-1 and NGAL(P＜0.05),an increase in pathological score of renal tubular injury(P＜0.05),increase in macrophage infiltration(P＜0.05),and an upregulation in the expression of M1-type macrophage-related molecules IL-6,IL-1β,TNF-α and iNOS(P＜0.05),as well as obviously increase in the percentage of M1-type macrophages as indicated by flow cytometry(P＜0.05),while there was no significant change in the percentage of M2-type macrophages,and the proportion of M1-type macrophages was significantly higher than that of M2-type macrophages(P＜0.05).Conclusion Fgl2 gene knockout exacerbates Cis-AKI by promoting macrophage polarization towards the M1 phenotype.

Objective To investigate the molecular mechanism of endothelial-mesenchymal transition (EndMT) induced by uremic toxin,parathyroid hormone (PTH),in vascular endothelial cells.Methods PTH (1 × 10-8 mol/L) was used to induce EndMT in human aortic endothelial cells (HAECs).TGF-β signaling inhibitor,including SB431542 and pirfenidone (PFD),and integrin-linked kinase (ILK) inhibitor Cpd22 were used to investigate the potential mechanism of EndMT induced by PTH in HAECs.Then the vascular endothelial cell markers VE-cadherin and CD31,and the mesenchymal marker α-SMA were detected by western blot.Results PTH reduced the expression levels of vascular endothelial cell marker CD31 and VE-cadherin (P＜0.05),while significantly increased the expression level of fiber cell marker α-SMA(P＜0.05).Furthermore,the TGF-βsignaling inhibitors (SB431542 and PFD) and ILK inhibitor (Cpd22) were able to partially reverse the EndMT induced by PTH in HAECs,which reversed the effect of PTH on reducing vascular endothelial cell marker expression and increasing fiber cell marker expression.Conclusion PTH could induce EndMT in HAECs via TGF-β/ILK pathway.