Objective    To evaluate the effect of gender matching on the outcomes of living-donor renal transplantation.    Methods    A total of 419 cases of living-donor renal transplantation in our center were divided into male-donor-male-recipient (MDMR) group, male-donor-female-recipient (MDFR) group, female-donor-male-recipient (FDMR) group, female-donor-female-recipient (FDFR) group. The outcomes including graft and patient survival, acute rejection and renal function were analyzed retrospectively.    Results    Compared to MDMR group, MDFR group and FDFR group had lower Scr [(80.7±17.9), (87.4±21.9) μmol/L vs (120.3±72.5) μmol/L, all P＜0.05] and uric acid (UA) [(318.1±86.4), (303.5±66.9) μmol/L vs (358.4±77.8) μmol/L, P＜0.05] 6 months after operation. Compared to MDFR group, FDMR group had higher Scr[(117.7±27.4) μmol/L vs (80.7±17.9) μmol/L, P＜0.01], UA [(371.0±92.4) μmol/L vs (318.1±86.4) μmol/L, P＜0.05] and lower glomerular filtration rate (GFR) [(70.4±17.8) ml/min vs (79.6±18.9) ml/min, P＜0.05]. Compared to FDMR group, FDFR group had lower Scr [(87.4±21.9) μmol/L vs (117.7±27.4) μmol/L, P＜0.01] and UA [(303.5±66.9) μmol/L vs (371.0±92.4) μmol/L, P＜0.01].  Compared to MDFR group, FDFR group showed lower GFR [(72.4±25.3) ml/min vs (82.7±18.7) ml/min, P＜0.05] 1 year after operation. Compared to MDMR group, FDFR group showed lower UA [(322.9±69.7) μmol/L vs (376.0±66.2) μmol/L, P＜0.05] 2 years after operation. Compared to FDMR group, FDFR group showed lower Scr [(88.7±27.0) μmol/L vs (112.7±27.8) μmol/L, P＜0.05] and UA  [(318.3±61.2) μmol/L vs (396.2±100.3) μmol/L, P＜0.05] 3 years after operation. 5 years after operation, there were no significant  differences in above indexes, the incidence of slow graft function, acute rejection and  survival of graft and patient among groups.    Conclusions    Male recipients of female donors have the worst renal function while female recipients have better outcomes after operation.

Objective    To evaluate the clinical efficacy of tacrolimus and cyclophosphamide (CTX) on lupus nephritis. Methods    The clinical trials on treatment of lupus nephritis with tacrolimus and CTX published before October 2012 were searched at Cochrane library, PubMed, OVID, Wanfang database, Chinese Journal full-text Database, Chongqing Weipu Database by using the methods of Cochrane systematic review. At the same time the information from related journals, professional data and network were hand-searched．The homogeneous evaluation was performed by Meta-analysis. Statistical analysis of clinical data was performed by using Stata11 software.    Results    A total of 4783 reports were found, while only 9 papers (8 randomized controlled trials and 1 cohort study) met the inclusion criteria. Tacrolimus group got better complete remission ratio (Z＝4.05, P＜0.01), similar partial remission ratio (Z＝0.44, P＝0.661), and better overall remission ratio (Z＝4.29, P＜0.01) as compared with CTX group. There were no significant differences between tacrolimus  group and CTX group in the incidence of infection (Z＝1.75, P＝0.081), renal damage (Z＝0.88, P＝0.38) and abnormal glucose metabolism (Z＝1.91, P＝0.070). Side effects such as liver function impairment (Z＝2.65, P＜0.01), gastrointestinal discomfort (Z＝2.31, P＜0.05), menstrual disorders (Z＝3.88, P＜0.01), bone marrow suppression (Z＝3.29, P＜0.01) in tacrolimus group were lower than those in CTX group.    Conclusions     Compared with CTX, tacrolimus has better complete remission ratio and overall remission ratio, with less side-effects in the treatment of lupus nephritis. However, large scale, multicenter, well-designed clinical trials should be adopted to further confirm the conclusions.
 

Objective    To investigate the epidemiological characteristics of hemoglobin variability (Hb-Var) in dialysis patients, and to explore the factors related to Hb-Var and the relationship between Hb-Var and patients outcomes.    Methods    The study enrolled 178 hemodialysis and peritoneal dialysis patients in the Department of Nephrology, Peking University People's Hospital between October 2009 and March 2010. First six months were observation period, and then followed up for 12 months. The Hb-Var was described by residual standard deviation (Res-SD) and fluctuation across threshold of the Hb level. Non-fatal cardiovascular events and mortality were the primary endpoints.    Results    The Res-SD was (4.74±2.66) g/L in dialysis patients. Most patients fell into groups of high amplitude (HA) and low amplitude low (LAL), with frequency of 29.8% and 33.1% respectively. The related factors of Hb-Var were fluctuation of erythropoietin (EPO) dosage and hemodialysis. Kaplan-Meier survival curve and multivariable adjusted Cox regression indicated that  Res-SD had no impact on the primary endpoints. Accumulative time to reach target hemoglobin level was an independent factor related to outcome.    Conclusion    Hb-Var occurs commonly in dialysis patients. The fluctuation of EPO dosage and dialysis style increase Hb-Var. Accumulative time to reach target hemoglobin level is an independent factor related to patients outcomes.

Objective    To investigate the prevalence and risk factors of chronic kidney disease (CKD) in the rural adult population of Zhuang nationality from the Guangxi Zhuang Autonomous Region.    Methods    A cross-sectional survey in a village of Zhuang agglomerated settlement was performed by cluster sampling. Demography data of participants were collected using questionnaire. Kidney damage indexes were examined and risk factors were explored. Morning spot urine albumin to creatinine ratio ≥30 mg/g was defined as albuminuria. Estimated glomerular filtration rate (eGFR) by abbreviated MDRD equation ＜60 ml•min-1•(1.73 m2)-1 was defined as reduced renal function. Morning spot urine dipstick (1＋or greater) and then ＞3 red blood cells/HP by microscopy was defined as hematuria. The crude and adjusted prevalence of indicators of kidney damage were calculated and risk factors associated with the presence of CKD was analyzed by Logistic regression.    Results    A total of 2104 subjects older than 18 years were enrolled in the study, of whom 2036 persons agreed and completed. After adjustment for age and gender, the prevalence of albuminuria, haematuria and reduced eGFR was 1.1%, 3.4% and 2.2%, respectively. The prevalence of chronic kidney disease was 5.7％ and the awareness rate was 5.3％. Independent risk factors associated with CKD were age, gender and hypertension.    Conclusions    In the Zhuang nationality village of Guangxi Zhuang Autonomous Region, 5.7％ people have either proteinuria, haematuria and/or reduced eGFR, indicating the presence of kidney damage, with awareness rate of only 5.3%. Independent risk factors associated with chronic kidney disease are age, gender and hypertension, which are similar to developed countries and domestic big cities.

Objective    To investigate the migration of bone-marrow mesenchymal stem cells (BMSCs) under acute kidney injury (AKI) microenvironment in vitro and the effect of erythropoietin (EPO) intervention, and to explore its underlying mechanism.    Methods    Renal tubular epithelial cells (RTECs) were cultured in hypoxia/ re-oxygenation (HR) condition for 12 h, respectively, in order to establish HR-RTEC. BMSCs and RTECs were co-cultured by Transwell system and were divided into 7 groups: control group (group①, only BMSC cultured), BMSC-RTEC co-culturing group (group②), BMSC-HR-RTEC co-culturing ＋EPO intervention groups (group③to group⑦, EPO concentration: 0, 1, 5, 10, 50 IU/ml). All the groups were cultured for 48 h and the number of migrating BMSCs was detected. Western blotting was applied for the detection of SDF-1 expression in RTECs and p-MAPK and MAPK levels in BMSCs. SDF-1 concentration in the RTECs culture supernatant was tested by ELISA.    Results    The number of BMSCs migrating to the low chamber where HR-RTECs were cultured was increased, and EPO intervention further enhanced this migration which reached the peak at the concentration of 10 IU/ml [Compared with group③, (46.67±7.37) cells vs (19.00±2.37) cells, P＜0.05]. Intracellular expression level and the secreated level of SDF-1 in HR-RTECs in group③ were higher than those in RTECs of group② [0.37±0.01 vs 0.19±0.01, P＜0.05; (61.64±4.88) μg/L vs (35.26±8.78) μg/L, P＜0.05]. EPO intervention increased above SDF-1 levels and reached the peak at the concentration of 10 IU/ml [group⑥ vs group③：(173.53±14.66) μg/L vs (61.64±4.88) μg/L, P＜0.05], accompanied with enhanced phosphorylation of MAPK in BMSCs.    Conclusions    AKI microenvironment has obvious chemotaxis effect on BMSCs, and EPO intervention can strengthen this effect. The increased SDF-1 level and enhanced phosphorylation of MAPK, the downstream signal protein of SDF-1/CXCR4 axis, are the possible mechanism for EPO performance.

Objective    To observe the effect of silibinin on the expression of integrin linked kinase (ILK), transforming growth factor β1 (TGF-β1) and α-smooth muscle actin (α-SMA) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose.    Methods    RPMCs were isolated, cultured and passaged by trypsin, then identified. The second generation of cultured RPMCs were used in the experiment. RPMCs were divided into normal control group, high glucose(1.5%, 2.5%, 4.25%) for 24 hours, high glucose (2.5%) for 12, 24, 48, 72 hours, high glucose (2.5%) for 24 hours after silibinin (5, 10, 20 mg/L) preincubate for 2 hours. ILK and α-SMA mRNA were detected by real-time PCR. ILK protein was detected by Western blotting. TGF-β1 protein in supernatants was detected by ELISA.    Results    Compared with the control group, the expresssion of ILK, TGF-β1 and α-SAM was significantly increased in groups stimulated by high glucose (all P＜0.05). Silibinin could significantly decrease the expression of ILK, TGF-β1 and α-SMA induced  by high glucose (all P＜0.05).      Conclusions    High glucose can up-regulate the expression of ILK, TGF-β1 and α-SMA. Silibinin can reverse these changes.

Objective    To investigate the effect of cysteine-rich protein 61 (Cyr61) on proliferation and cell cycle in human renal tubular epithelial cells (HK-2).    Methods    Cyr61 cDNA was cloned into pEGFP-N2, then HK-2 cells were transfected with the recombinant plasmid pEGFP-N2-Cyr61 by Lipofectamine. The cell proliferation was measured by MTT. The expression level of Cyr61,    p-FAK and cyclin dependent cyclin-dependent kinase 2 (CDK2) protein were detected by Western blotting. The cell cycle and cell apoptosis were analyzed by flow cytometry.    Results    The recombinant plasmid pEGFP-N2-Cyr61 could be transfected into HK-2 efficiently. After transfection, the proliferative activity was significantly increased, the proportion of HK-2 cells in G1 phase decreased and in S-phase increased significantly, the level of cell apoptosis decreased markedly (all P＜0.01). The expressions of Cyr61, p-FAK and CDK2 in Cyr61-transfected group were all amplified significantly (all P＜0.01).    Conclusions    Cyr61 protein over-expressed in HK-2 cells can increase CDK2 expression throngh FAK pathway, resulting in the promotion of HK-2 cells entering into S phase, cell proliferation and the reduction of cell apoptosis.
 

Objective    To explore the renoprotective effect of N-acetylcysteine (NAC) on chronic cyclosporine A (CsA) nephrotoxicity in mice model.    Methods    ICR mice were randomly      divided into 4 groups: normal control group [olive oil, 1 ml/kg subcutaneous injection (s.c)], NAC control group (olive oil, 1 ml/kg s.c plus NAC 150 mg•kg-1•d-1 in drinking water), model group        [ICR mice maintained on a salt-depleted diet (0.05% sodium) plus CsA (30 mg•kg-1•d-1) for four weeks]; Treatment group [ICR mice maintained on a salt-depleted diet (0.05% sodium) plus CsA (30 mg•kg-1•d-1 s.c) and NAC (150 mg•kg-1•d-1 in drinking water). Basic parameters, histopathology,    8-hydroxydeoxyguanosine (8-OHdG), the expression of Klotho and AKT-FoxO1 were studied.    Results    Compared with the normal control group, mice in model group showed deterioration in renal function [Scr (27.8±1.2) μmol/L vs (19.0±2.2) μmol/L, P＜0.01], development of tubulointerstitial fibrosis, increased urinary 8-OHdG output [(41.2±16.8) ng/d vs (28.7±7.4) ng/d, P＜0.05], and decreased Klotho expression [(17.8±4.5)% vs (100.0±4.0)%, P＜0.01]. Concomitant administration of NAC significantly improved all above parameters (all P＜0.05). Correlation analysis revealed that Klotho expression was negatively correlated with urinary 8-OHdG excretion (r＝-0.934, P＜0.01) and AKT-FoxO1 expression （AKT: r＝-0.939, P＜0.01; FoxO1: r＝-0.919, P＜0.01).    Conclusion    NAC can attenuate tubulointerstitial fibrosis in chronic CsA nephrotoxicity mice, which may be associated with the up-regulation of Klotho expression and the inhibition of AKT-FoxO1 signaling pathway.

Objective    To investigate the effects of triptolide on tubulointerstitial fibrosis in rats kidneys with the unilateral ureteral obstruction (UUO) by examining the expression of collagen type Ι (Col-Ι), Ski, Smad3, TGF-β1.    Methods    Sixty male SD rats were divided into three groups: Sham operation group (Sham group), UUO group and triptolide (0.2 mg•kg-1•d-1) treatment group. The levels of blood urea nitrogen (BUN), serum creatinine (Scr), pathological changes were measured. Col-Ι, Ski and Smad3 expressions were assessed by immunohistochemistry. Protein and mRNA expressions of Ski, Smad3, TGF-β1 were assessed by Western blotting and real-time PCR.    Results    Compared with Sham group, Scr and BUN increased significantly in UUO group (P＜0.05). Interstitial fibrosis was prominent and renal interstitial injury score increased significantly in UUO group (P＜0.05). The expressions of Col-Ι and Smad3 were increased in UUO group (P＜0.05). Compared with Sham group, the protein expressions of TGF-β1 and Smad3 were increased, the Ski protein was decreased in UUO group  (P＜0.05). In triptolide group, the morphological changes were notably reduced  (P＜0.05). Comparison with UUO group, triptolide could increase the protein and mRNA expressions of Ski significantly, and decreased the protein and mRNA expressions of Smad3 and TGF-β1  (P＜0.05).    Conclusion    Triptolide can reduce the tubulointerstitial fibrosis by up-regulating Ski, and down-regulating TGF-β1 and Smad3.

Objective    To investigate the molecular mechanism of aristolochic acid (AA)-induced renal tubular epithelial cells (NRK-52E) injury, and to elucidate possible role of sonic hedgehog (Shh) signaling in this process.    Methods    The proliferation inhibition rate was measured by WST-1 assay in vitro after NRK-52E cells were treated with AA (1, 10, 100 mg/L) for 12, 24, and 48 h, respectively. Cell apoptosis was determined by Hoechst 33258 staining. mRNA expressions of Smo, Ptch1, α-SMA, E-cadherin, TGF-β1, and type III collagen were detected by real-time PCR. The levels of Shh and TGF-β1 were detected by ELISA assay. The expressions of bcl-2, bax, α-SMA, and  E-cadherin were examined by Western blotting.    Results    WST-1 assay showed that AA significantly inhibited the proliferation of NRK-52E cells after 24 h, which was in a dose-dependent pattern (r＝0.817, P＝0.047). Evidence from Hoechst 33258 staining revealed that lower concentration (1, 10 mg/L) AA induced cell apoptosis, but higher concentration (100 mg/L) AA induced cell necrosis. In AA-treated cells (10 mg/L), apoptosis was induced, which was partly mediated by the mitochondrial pathway with decreased bcl-2 and enhanced bax expression. The over-expression of Shh protein and Smo mRNA, and down-regulation of Ptch1 mRNA expression were found, indicating that Shh signaling was activated. In addition, the expressions of α-SMA, TGF-β1, and type Ⅲ collagen were significantly increased, but E-cadherin expression was decreased, suggesting that epithelial-to-mesenchymal transition and fibrosis occurred.    Conclusions    AA can significantly inhibit proliferation, and induce apoptosis and necrosis in NRK-52E cells. In this process, Shh signaling is activated, which promotes epithelial-to-mesenchymal transition and fibrosis.

Objective    To investigate the effects of angiotensin Ⅱ(AngⅡ) stimulating on cholesterol influx in human renal proximal tubular epithelial cells (HK-2) and the relation to low-density lipoprotein receptor (LDLr) pathway.    Methods    HK-2 cells were cultured and divided into the control group (incubated with serum-free medium) and AngⅡ group (treated by 10-7 mol/L of AngⅡ for 24 hours). The effects of AngⅡ on lipid accumulation were examined by Oil red O staining and a quantitative assay of intracellular cholesterol. The expression of LDLr, sterol regulatory element-binding protein (SREBP) cleavage activating protein (SCAP) and SREBP-2 mRNA and protein were examined by real-time PCR and Western blotting. The cotranslocation of SCAP-SREBP-2 from endoplasmic reticulum to Golgi in HK-2 cells was examined by immunofluorescent staining under confocal microscopy.    Results    AngⅡ treatment increased intracellular lipid accumulation in HK-2 cells, which was associated with increased mRNA and protein expression of LDLr, SCAP, and SREBP-2 in HK-2 cells induced by AngⅡ. Furthermore, results from confocal microscopy observation demonstrated that AngⅡ increased the translocation of SCAP/SREBP-2 complex from endoplasmic reticulum to Golgi, thereby up-regulating LDLr gene transcription.    Conclusion    AngⅡ disrupts LDLr feed-back regulation to increase cholesterol uptake and induce intracellular lipid accumulation.