Objective To observe the fine distribution and morphologic feature of the intrarenal lymphatics. Meth-ods 20 Wistar rats were involved in this study and nephridial tissues were obtained from each animal. Double im-munofluorescence was applied to detect the lymphatic distribution by using Podoplanin and CD34. Renal tissues were obtained from the cortex, medulla and junctional zone,and put in 3% glutaric dialdehyde stationary liquid for transmission electron microscope to observe the fine distribution and morphologic feature of the intrarenal lymphat-ics. Results Under immunofluorescence double staining, lymphatic capillary appeared anomalism, large lumina, thin wall and located in connective tissue region. Cortilymph obviously exceeded medulla lymph in rats. Under e-lectron microscope, the basal lamina of lymphatic was discontinuity. A lot of lysosome and vesicles existed in the endothelial of capillary lymphatics. Many of the vesicles liberated in cytoplasm. The intercellular junctions of the renal lymphatics were held in close apposition. Conclusion The intercellular junctions of the renal lymphatics are held in close apposition by adhesion devices. A lot of lysosomes and vesicles exist in the endothelial of capillary lymphatics, which are the base for renal to transport big molecules and fluids.

Objective To examine the association of pro-hepcidin with iron metabolism and inflammation in maintenance hemodialysis (MHD) patients with erythropoietin (EPO) resistance.Methods Forty MHD patients and twenty healthy controls were enrolled in the study.Among MHD patients,20 were hyporesponsive to EPO therapy and 20 were normal responsive to EPO therapy.Complete blood red cell count (RBC),Hb concentration,hematocrit (Hct),reticulocyte count (Ret),and serum ferritin (SF),serum iron (Fe),total ironbinding capacity (TIBC),saturation rate of transferrin (TSAT),transferrin (TF),hyper-sensitive C-reactive protein (hs-CRP),pro-hepcidin were measured in all the patients and controls.Differences were compared between groups.Influencing factors were analyzed by Pearson correlation.Predicting value of pro-hepcidin was investigated by ROC curve.Results Serum levels of SF,pro-hepcidin and hs-CRP were significantly higher in MHD patients than those in healthy controls (P＜0.01),while serum TF was lower in MHD patients (P＜0.05).Serum levels of SF,pro-hepcidin and hs-CRP were significantly higher in EPO resistant patients as compared to normal responsive cases (P＜0.01).Serum prohepcidin level was positively correlated with SF (r=0.843,P=0.000) and hs-CRP (r=0.695,P=0.001).In predicting EPO resistance,area under ROC curve of pro-hepcidin,SF and hs-CRP was 0.713,0.769 and 0.958 respectively.Conclusions EPO resistance is correlated with inflammation and iron metabolism.Serum pro-hepcidin,SF and hs-CRP may be used as markers of EPO resistance in MHD patients.

Aim To observe the effect of mycophenolate mofetil(MMF) on expressions of MCP-1 and CD68 in renal tubulointerstitial injury of diabetic rats and explore the mechanism of MMF′s protective role.Methods Diabetes was induced in uninephrectomized male Wistar rats by peritoneal injection of STZ (65 mg?kg-1). Rats were randomly divided into three groups:control group (NC),diabetic group (DM) and treated group (DM+MMF) with MMF(15 mg?kg-1?d-1).This study lasted for 8 weeks. 24 h urinary protein,blood glucose and the ratio of left kidney weight/body weight were determined after 8 weeks.The renal tubulointerstitial morphological change was observed,immunohistochemical method was used to analyze expressions of MCP-1 protein and CD68. Expression of MCP-1 mRNA in renal tissue was measured by quantitative Real-time PCR.Results Compared with NC group, serum glucose level,24 hour urinary protein and the ratio of left kidney weight/body weight were significantly increased(P

[ ABSTRACT] AIM: To investigate whether L-carnitine ( LC) inhibits the eryptosis effect of uremic serum on erythrocytes.METHODS:Erythrocyte suspension (2%) was cultured and divided into 3 groups in vitro: control group ( C group) , uremic serum group ( U group, 30%uremic serum) , and uremic serum+LC group ( L group, 30%uremic serum+200 μmol/L LC) .Erythrocytes were collected at 24 h and 48 h.Eryptosis ( phosphatidylserine expression repre-sents eryptosis) was estimated by flow cytometry with Annexin V staining.The content of reactive oxygen species ( ROS) was also detected.Glutathione ( GSH) was measured by ELISA.RESULTS:Eryptosis in C group was increased as the in-cubating time extended.Eryptosis in U group was higher than that in C group, while that in L group was lower than that in U group.Meanwhile, ROS content was higher and GSH was lower in U group than those in C group.ROS content was low-er and GSH was higher in L group than those in C group.CONCLUSION:LC inhibits uremic serum-induced eryptosis by decreasing ROS and increasing GSH, thus attenuating oxidative stress.

HBx gene is a multifunctional regulator,which has extensive trans-activating effects,and can activate transcription factors,inhibit DNA repair and regulate cell proliferation,differentiation and apoptosis.In recent years,the role of HBx gene in pathogenesis of hepatitis B virus-associated glomemlonephritis (HBV-GN) has been extensively studied,and the results show that HBx can promote glomerular mesangial cell proliferation,and induce damage or apoptosis of podocytes and renal tubular epithelial cells.This paper reviews the research progress on biological characteristics of HBx and its role in pathogenesis of HBV-GN.

Objective To determine whether mutation of Hepatitis B virus (HBV) X gene is associated with hepatitis B virus-associated glomerulonephritis (HBV-GN).Methods The venous blood was collected from 50 patients with HBV-GN and 60 patients with asymptomatic HBV carriers (control group).Serum HBV DNA was extracted to determine the serum titer of HBV-DNA and then polymerase chain reaction (PCR) was used to detect the HBV X gene mutation.Results (1)There were not statistical significance between age and gender in HBV-GN group and control group (P ＞0.05).There were not statistical significance of serum replication level of HBV DNA in HBV-GN with X gene mutation and control group (P ＞ 0.05).Urine protein excretion in HBV-GN group with or without X gene mutation was found with statistical significance (P ＜ 0.05).(2)Nucleotide mutations [84％ (42/50)] resulted in amino acid substitution in HBV-GN.Nucleotide mutations changed in transfunction control region of X gene,including position nt1653,nt1726,nt1727,nt1730,nt1753,nt1762 and nt1764.(3)Nucleotide mutations [8％(5/60)] resulted in amino acid substitution in control group.Nucleotide mutations changed in position nt1632 and nt1635,located in non-functional region.Conclusions HBV X gene mutations and the subsequent amino acid substitutions are found in HBV-GN.The urine protein excretion level increases in patients with X mutation,suggesting that these mutations may play an important role in the pathogenesis of HBV-GN.

Objective To investigate the correlation of HBV DNA level with clinicopathological characteristics and prognosis of HBV-associated glomerulonephritis ( HBV-GN ) .Methods One hundred and two patients with HBV-GN admitted in Affiliated Hospital of Qingdao University during January 2009 and October 2012 were successively enrolled.Fluorescence quantitative polymerase chain reaction was used to determine the serum titer of HBV DNA.According to HBV DNA levels the patients were divided into three groups:low-replication group (HBV DNA <103 copies/mL), moderate-replication group (103 copies/mL ≤HBV DNA≤105 copies/mL) and high-replication group ( HBV DNA >105 copies/mL) .Renal biopsy was performed to determine the pathological type, and immunofluorescence assay was used for quantitative detection of HBV related antigens ( HBsAg, HBcAg and HBeAg ) in kidney. All patients received lamivudine (100 mg/d) plus adefovir dipivoxil (10 mg/d) combination therapy and followed up for 18 months.The renal function, biochemical and immunological indexes before and after the treatment were measured.One-way ANOVA was used to analyze the differences of above parameters among patients in three groups.Spearman correlation coefficient was used for correlation analysis between HBV DNA level and pathological stages in kidney.Results There were 20 patients in low-replication group, 51 in moderate-replication group and 31 in high-replication group.With the increase of serum level of HBV DNA, 24-h urine protein excretion, plasma cholesterol, and triglycerides increased (F=34.64, 40.10 and 31.72, P<0.01), plasma level of albumin decreased (F=24.04, P<0.01), and the immune complexes of HBV related antigens ( HBsAg, HBcAg and HBeAg) in kidney increased ( F=41.49, 15.64 and 10.41, P<0.01).For 78 patients with HBV-membranous nephropathy ( HBV-MN), the pathological injury was aggravated with the increase of serum level of HBV DNA (r=0.38, P<0.01).The level of 24-h urine protein excretion declined after treatment in three groups ( t =7.86, 19.28 and 16.74, P <0.01 );complement C3 increased, but no statistical significance was observed ( t =1.05, 1.04 and 1.94, P >0.05);no change in creatinine was found (t =0.14, 0.52 and 0.57, P >0.05).After 18-month treatment, clinical remission rates in three groups were 95.0% ( 19/20 ) , 70.6% ( 36/51 ) and 54.8%(17/31), respectively.The clinical remission rate was significantly lower in the high-replication group as compared with that in low-replication group (χ2 =9.44, P<0.01).Conclusion Serum level of HBV DNA is closely correlated with renal function, renal pathology and clinical remission rate in HBV-GN patients, which may be used for the evaluation of kidney biopsy, treatment and prognosis in patients with HBV-GN.

OBJECTIVE: To explore the effects of Salvia miltiorrhiza on renal morphology and renal function of rats with streptozotocin diabetes. METHODS: Thirty male Wistar rats were randomly divided into three groups, which were normal control group, untreated group and Salvia miltiorrhiza-treated group. Diabetic nephropathy was induced in rats of the last two groups by intraperitoneal injection of streptozotocin after unilateral nephrectomy. Then the rats in the normal control and untreated groups were fed with normal saline while those in the Salvia miltiorrhiza-treated group were fed Salvia miltiorrhiza preparation for 8 weeks. The glomerular volume (VG), kidney-to-body weight ratio (KW/BW), urinary albumin excretion rate (UAER) and creatinine clearance (Ccr) were observed. The expression levels of transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF), fibronectin (FN) and plasminogen activator inhibitor-1 (PAI-1) were detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) at the end of the experiment. RESULTS: UAER, Ccr, VG and KW/BW ratio were significantly higher in the untreated group than those in the normal control group (P<0.05). The expression levels of TGF-beta1, CTGF, PAI-1 and FN in the untreated group were also significantly higher as compared with those in the normal control group (P<0.05). UAER, Ccr, VG, KW/BW ratio and the levels of TGF-beta1, CTGF, PAI-1 and FN in the Salvia miltiorrhiza-treated group were obviously lower than those in the untreated group (P<0.05). CONCLUSION: Salvia miltiorrhiza can protect rats with streptozotocin diabetes from diabetic nephropathy by suppressing the over-expressions of TGF-beta1, CTGF, PAI-1 and FN in renal cortex.

Objective To explore the effects of human umbilical cord mesenchymal stem cells (HUC?MSCs) on podocytic apoptosis and injury induced by high glucose (HG) and the underlying mechanisms. Methods Podocytes were divided into six groups according to treatment: ⑴ normal glucose group (NG);⑵high glucose group (HG);⑶mannitol control group (NG+Ma);⑷HUC?MSC co?culture group (HUC?MSCs); ⑸ recombinant human hepatocyte growth factor treatment group (rhHGF);⑹ neutralizing antibody group(HGF?NtAb). Cytometry and Hoechst staining were used to detect the apoptosis rates. Western blot was used to measure the ratio of active PARP to total PARP and the level of Bcl?2. Immunofluorescence was used to study podocytic apoptosis and injury. Neutralizing antibody (NtAb) was used to block its function and the recombinant cytokine was added to induce its function. Results High glucose induced podocytic apoptosis in a time?dependent manner, HUC?MSCs co?culture decreased the podocytic apoptosis rate and the expression of PARP (all P﹤0.05), increased the expression of Bcl?2, prevented the reduced expression and maintained the normal arrangement of podocytic podoplanin. The rhHGF prevented podocytic apoptosis and injury similarly to HUC?MSCs, the beneficial effect of HUC?MSC decreased when blockade of HGF. Conclusions HUC?MSCs co?culture ameliorates podocytic apoptosis and injure induced by HG, probably through secreting soluble HGF.

Objective To examine the correlation of imbalance of urinary exosome Th1/Th2 with diabetic nephropathy (DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) and 30 healthy volunteers as control were enrolled in the study.According to urinary albumin/creatinine ratio (UACR),type 2 diabetes mellitus patients were divided into 3 groups:diabetes mellitus without nepbropathy group (DM,n=40,UACR＜30 mg/gCr),microalbuminuria group (DN 1,n=50,UACR-30～300 mg/gCr) and clinicoalbuminuria group (DN 2,n=30,UACR＞300 mg/gCr).Urine exosome-interferon-gamma (IFN-γ) and exosome-interleukin 4 (IL-4) levels were determined by enzyme-linked immunosorbent assay (ELISA).Multiple stepwise linear regression was used to analyze the correlation of exosome-IFN-γ/IL-4 with glycated hemoglobin (HbA1c),cholesterol (CH),UACR,Scr and BUN.Results Th1/Th2 ratio in DM,DN1,DN2 groups was significantly higher than that in healthy group (0.8089±0.2458,0.8993 ±0.3515,0.8571±0.2470 vs 0.6198±0.1769,all P＜0.01).Correlation analysis showed that urinary exosomeIFN-γ/IL-4 ratio was positively correlated with UACR (r=0.213,P=0.015) and BUN (r=0.292,P=0.001).Multiple stepwise linear regression analysis showed that BUN was independent determinants for exosome-IFN-γ/IL-4 (β=0.246,P=0.006).Conclusion The imbalance of urinary exosomeTh1/Th2 is correlated with DN,which may play an important role in the pathogenesis of DN.