Objective To analyze the distribution and antimicrobial resistance of pathogenic bacteria in urinary tract infection (UTI) so as to provide evidence for appropriate selection of antimicrobial agents in clinical practice. Methods From January 2001 to December 2008 in Shanghai Ruijin Hospital, 4683 strains of pathogenic bacteria isolated from urine samples were detected by ATB system; drug susceptibility test was performed with disk diffusion method and pathogenic bacteria distribution and drug resistance was analyzed with WHO NET 5.3 software. Results Among 4683 strains of pathogenic bacteria, most was gram-negative bacilli, accounting for about 77.8%, of which predominant strain was Escherichia coli (68.7%, 3217/4683). The predominant strain of gram-positive bacteria was Enterococcus faecalis, accounting for 10.0% (468/4683). Escherichia coli showed high resistance rates to ampicillin, piperacillin and compound sulfamethoxazole (SMZ-TMP), which were 76.6%, 61.7% and 57.4% respectively, while a low resistance to imipenem, cefoperazone-sulbactam, piperacillin-tazobactam. Enterococcus faecalis showed high resistance rates to erythromycin, gentamicin and levofloxacin, which were 65.8%, 43.2% and 31.1% respectively, and were most susceptive to vancomycin and teicoplanin, both with resistance rates of 0. The susceptibility rate of Enterobacteriaceae to imipenem was 100%. From 2006 to 2008, the detection rate of extend-spectrum ?茁-lactamases ESBLs -producing Escherichia coli in outpatient increased year by year, from 28.7% to 43.3% (P<0.05), whereas no significant change was found in inpatients. The detection rate of (ESBLs)-producing Escherichia coli in inpatients was significantly higher than that in outpatients (P<0.05). The detection rate of ESBLs-producing Escherichia coli was 23.6%. The antimicrobial resistance rate in elderly patients was significantly higher than that in overall antimicrobial resistance rate (P<0.05). Conclusions The predominant bacteria of UTI are still gram-negative bacteria, main of which is Escherichia coli. Bacteria are resistant to a variety of antibiotics. Approximate selection of antibiotics in clinical practice should be made on the basis of susceptibility test results.

Objective To assess the incidence, risk factors and mortality of acute kidney injury (AKI) in patients with chronic myelogeneous leukemia (CML) after myeloablative allogenetic hematopoietic stem cell transplantation (HSCT). Methods Renal function in 93 CML patients undergone myeloablative allo-HSCT was retrospectively analyzed by the RIFLE criteria. Results Thirty-nine patients(41.9%) developed AKI at a median of 40 days after allo-HSCT, including 24 AKI-R patients(25.8%)，10 AKI-I patients(10.8%) and 5 AKI-F patients(5.4%). The morbidity of AKI in patients with ≥Ⅲ acute graft-versus-host disease (aGVHD) and without <Ⅲ GVHD was (81.82±11.63)% and (36.59±5.32)%(P＝0.0037)respectively. The morbidity of AKI in patients with increased total bilirubin and without increased total bilirubin was (72.73±13.43)% and(37.04±5.37)%(P＝0.0192) respectively. ≥ⅢaGVHD was poor-prognostic factor of AKI and RR was 2.773[95%CI (1.073-7.167), P＝0.035]. RR of AKI-I and AKI-F in patients with ≥ⅢaGVHD was 6.320[95%CI (1.464-27.291), P＝0.013]. The mortality within 100 days after allo-HSCT of patients with AKI was significantly different as compared to patients without AKI (P＝0.001). Six-month survival rates of different class AKI patients after myeloablative allo-HSCT were (86.96±7.02)% (AKI-R), (70.00±14.49)% (AKI-I), 0 (AKI-F) (P＝0.000) respectively. Conclusions AKI is one of the main complications in CML patients after myeloablative allo-HSCT. ≥ⅢaGVHD and increased total bilirubin are poor-prognostic factors of AKI, and higher morbidity of AKI-I and AKI-F can be found in patients with ≥ⅢaGVHD. With the deteriorated AKI, 6-month survival is decreased. RIFLE criteria is sensitive to the early diagnosis of renal function. Moreover RIFLE can monitor the progression of AKI and predict the clinical outcome.

Objective To assess the safety and efficacy of gabapentin in treatment of refractory uremic pruritus in maintenance hemodialysis (MHD) patients. Methods A randomized controlled trial was performed. Forty-nine MHD patients with severe pruritus were randomly divided into treatment group (gabapentin 100-300 mg at dialysis day before sleep) and the control group (loratadine 10 mg daily). The efficacy was assessed by visual analogue score, pruritus score and improved Duo’s VAG score after 12-week treatment, and the side effect was also observed to assess the safety. Results After treatment, the frequency, degree and area of pruritus in 25 patients of treatment group were significantly reduced, and VAS score (1.46±1.38 vs 8.71±1.17, P<0.01), VAG score (2.92±1.63 vs 8.29±0.68, P<0.01) and the improved Duo’s pruritus score (11.33±3.99 vs 30.75±4.87, P<0.01) decreased significantly compared with that of prior treatment. The symptoms of 24 patients in control group were improved partly, which were not as good as those in treatment group (P<0.01). The side effects of gabapentin were drowsiness and dizziness, but most symptoms were relieved or disappeared within 1 week and no patient interrupted therapy. No serious adverse events were observed. Conclusion Gabapentin is safe and effective for refractory uremic pruritus, but long-term efficacy and safety requires larger sample and long-term observation.

Objective To investigate the interrelationship among urinary albumin elimination rate (UAER), serum adiponectin (ADPN) and blood glucose level in patients with type 2 diabetic nephropathy(DN). Methods A total of 89 type 2 diabetic patients and 30 healthy people as control were enrolled in the study. According to UAER, the diabetic patients were divided into three groups: normal group (35 cases, UAER<30 mg/24 h), microalbuminura group (32 cases, UAER 30-300 mg/24 h) and macroalbuminura group (22 cases, UAER>300 mg/24 h). Serum adiponectin was measured by ELISA. Body mass index (BMI), waist hip radio (WHR), fasting plasma glucose (FPG), 2-hour plasma glucose (2hPG), glycated haemoglobin (HbAlc), blood lipid and UAER were measured routinely. Data were compared among groups. Results Serum adiponectin level was lower in diabetic patients than that in healthy control group(P＜0.05). Among diabetic patients, serum adiponectin of macroalbuminura group was significantly higher than that of normal and microalbuminura groups [(67.74±14.89) vs (39.36±13.92), (54.38±10.14) ng/L, P＜0.05]. Multiple regression analysis showed 2hPG, HbA1c, BMI, disease course, UAER and DBP were closely associated with the level of serum adiponectin. Correlation analysis indicated serum adiponectin was positively correlated with age (r=0.255), disease course (r=0.405), HDL (r=0.501) and UAER (r=0.843); while negatively with HbA1c (r=-0.479), FPG (r=-0.436), 2hPG(r=-0.418), BMI (r=-0.479) and WHR (r= -0.531), all P＜0.01. Conclusions Serum adiponectin is positively correlated with UAER and negatively correlated with FPG. Blood glucose level is one of the main factors affecting serum adiponectin. Strict controlling of blood glucose level is beneficial to higher level of ADPN.

Objective To explore the effect of serum uric acid (SUA) on the clinicopathological manifestation and prognosis of IgA nephropathy (IgAN) patients. Methods A total of 348 patients with renal biopsy-proven IgAN in our hospital were enrolled in this study. The data were retrospectively analyzed to examine the association of SUA level with clinicopathological manifestation and prognosis of IgA nephropathy (IgAN) patients. Results There were no significant differences of 24 hour proteinuria, BUN and Scr between patients of high SUA level with various GFR and those of normal SUA level. While differences of glomerular sclerosis, tubulointerstitial scores and vascular injury between these two groups were significant (P＜0.05). At the end of follow-up, prevalence of GFR decline and ESRD was significantly higher in patients with high SUA as compared to those with normal SUA (40.82% vs 15.70%, 64.71% vs 35.00%, respectively, P＜0.05). Conclusions Patients with different SUA levels have similar clinical manifestations, but different pathological findings and prognosis. It is important to pay attention to the follow-up of SUA level in IgAN patients.

Objective To study the effect of ERK signalling pathway and aldose reductase (AR) on the transforming growth factor-β1 (TGF-β1)-induced expression of fibronectin (FN) in nondiabetic nephropathy. Methods Human mesangial cells (HMCs) were cultured and transfected with pCDNA3-AR and AR gene silencing with small interfering RNA (siRNA). The AR expression in the HMCs was examined by real-time PCR and Western blotting was used to detect the protein expression of AR and FN. Inhibitors of AR and ERK signalling pathway were co-cultured with HMCs, then TGF-β1 was added and Western blotting was used to analyze the protein expression of FN. Results The expression of AR, FN and p-ERK was up-regulated by TGF-β1. AR was increased by 1.8-fold, FN was increased by 1.9-fold (P<0.05), and p-ERK was increased by 5.1-fold after stimulation with TGF-β1 (P<0.01). HMCs transfected with AR showed stronger protein expression of FN, more than 3.6-fold in the protein level of FN was observed in HMCs (P<0.05). The HMCs of knockdown AR gene by siRNA showed decreased expression of AR, FN and p-ERK, the level of AR mRNA in HMCs transfected with AR siRNA was 10% of the level in untransfected cells or cells transfected with control siRNA (P<0.01). Transfection with AR siRNA attenuated TGF-β1-induced FN production, more than 70% decrease in the protein level of FN and p-ERK was observed in HMCs with AR-siRNA (P<0.01). Conclusions AR can regulate the expression of FN with the stimulation of TGF-β1, as AR gene is one of the responsive genes of TGF-β1, which may be associated with the activation of ERK signalling pathways induced by TGF-β1.

Objective To investigate the effect of suppressor of cytokine signaling-1 (SOCS-1) on expression of monocyte chemoattractant protein-1 (MCP-1) in human glomerular mesangial cells(HMCs) under high concentration of glucose. Methods Stable transfections of HMC with pCR3.1 vector and pCR3.1-SOCS-1 were performed with lipofectamine 2000, and cells were selected with geneticin. Cells were stimulated with low glucose（LG, 5.5 mmol/L）, high glucose (HG, 30 mmol/L), LG plus mannitol (24.5 mmol/L) and AG490 (10 μmol/L). The protein expression levels of SOCS-1, signal transducer and activators of transcription 1,3 (STAT1, STAT3), p-STAT1 and p-STAT3 were observed by Western blotting. The protein synthesis of MCP-1, FN and type Ⅳ collagen in the supernatants of the HMCs were detected by ELISA and radioimmunoassay. The expression level of SOCS-1 and MCP-1 mRNA was measured by RT-PCR. Results HG induced the expression of SOCS-1 protein and mRNA in HMCs in time-dependant manner, peaked at 4 h, and gradually decreased to baseline at 24 h. Compared with low glucose control group, the phosphorylation levels of STAT1 and STAT3 and the expression of MCP-1 mRNA [(0.39±0.05) vs (0.16±0.02)] were significantly increased in HMCs under high glucose medium (P <0.01）. Exposure of HMCs to high glucose conditions showed high concentration of MCP-1[(459±67) ng/L vs (241±19) ng/L], fibronectin [(5.84±0.61) mg/L vs (3.41±0.31) mg/L] and type Ⅳ collagen [(16.45±2.30) μg/L vs (9.56±1.52) μg/L] in the supernatants(all P<0.01). Overexpression of SOCS-1 inhibited the phosphorylation levels of STAT1 and STAT3 and the expression of MCP-1 mRNA [(0.34±0.04) vs (0.42±0.05)] in HMCs under high glucose condition(all P<0.05). Compared with vector control group, the concentration of MCP-1[(387±47) ng/L vs(463±56) ng/L], fibronectin [(4.61±0.57) mg/L vs (5.76±0.74) mg/L] and type Ⅳ collagen [(13.4±2.32) μg/L vs (17.1±2.57) μg/L] was decreased in supernatants of HMCs with SOCS-1 overexpression(all P<0.05). Compared with HG group, the expression of MCP-1 mRNA (0.31±0.04) and the concentration of MCP-1 [(361±53) ng/L], FN[(5.46±0.71) mg/L] and type Ⅳ collagen [(15.2±1.97) μg/L] in supernatants were decreased in HMCs treated with AG490. Conclusion Overexpression of SOCS-1 inhibits overproduction of MCP-1 and ECM proteins in HMCs under high glucose conditions, which may be partly by regulating the phosphorylation of STAT1 and STAT3.

Objective To investigate the effect of 4-phenylbutyric acid(4-PBA) on the renal pathogenesis of rats with streptozotocin-induced diabetes and its mechanism. Methods Fifty-four male SD rats were randomly divided into three groups: normal control group (NC group, n= 18), diabetic nephropathy group (DN group, n=18), diabetic nephropathy plus 4-PBA treatment group (4-PBA group, n=18). At the end of 4, 8 and 12 weeks, index of kidney weight/body weight ratio (KI) were measured and calculated. Serum creatinine (Scr), blood urea nitrogen (BUN), urinary MDA levels, urinary SOD activity, and 24 hour urinary protein excretion rate (UAER) were detected by HITACHI automatically. Morphology of kidney was examined by special staining of periodic acid-schiff (PAS). The p47phox and nitrotyrosine (NT) expression in kidney were determined by real-time fluorescence PCR and Western blotting. Results Compared with the NC group, the DN group rats showed a significant increase of KI(P<0.05), UAER(mg/24 h) (4.92±0.70 vs 0.26±0.07, 5.29±0.83 vs 0.28±0.08, 5.54±0.81 vs 0.29±0.04，respectively, P<0.05] for indicated time, mesangial cells proliferation and mesangial matrix expansion at 12 week. However, 4-PBA treatment could significantly inhibit the increase of KI(P<0.05), decrease UAER(mg/24 h) (3.71±0.37, 3.47±0.36, 3.28±0.40， respectively, P<0.05] for indicated time, and prevent the glomeruler pathological alteration induced by diabetes. Moreover, the mRNA expression of p47phox in the kidney of DN group was 154.72%, 148.60% and 91.95% more than that of NC group (all P<0.05) for indicated time. The protein expression of p47phox was 118.00%, 140.10% and 177.82% more than that of NC group (all P<0.05), and the protein expression of NT was 45.29%, 59.13% and 89.28% more than that of NC group(all P<0.05). In addition, urinary MDA levels in DN group were 2.05-, 2.26- and 2.43- folds of NC group, and urinary SOD activities were decreased by 64.78%, 71.29% and 79.32% of NC group. Compared with the DN group, the mRNA and protein expression of p47phox, and protein expression of NT in 4-PBA group were decreased markedly(all P<0.05) at the end of 8 and 12 weeks. The urinary MDA level was decreased, and the urinary SOD activity was increased significantly in rats with diabetes after 4-PBA treatment for indicated time (all P<0.05）. Conclusion 4-PBA treatment can significantly inhibit the renal pathogenesis of rats with diabetes through inhibition of oxidative stress.

Objective To explore the effect of aldosterone on renal epithelial-mesenchymal transition in streptozocin(STZ)-induced diabetic nephropathy rats. Methods Wistar rats were intraperitoneally injected with STZ (60 mg/kg) for the preparation of diabetic model. After 4 weeks, the rats with urinary protein> 30 mg/d were regarded as successful diabetic nephropathy(n=16), and were randomly divided into diabetic nephropathy (DN group, n=8) and spironolactone group (SP group, n=8). Then eight healthy rats were selected randomly as control group (N group, n=8). SP group rats were treated with spironolactone 40 mg&#8226;kg-1&#8226;d-1, and N group and DN group rats were given equal water. After 8 weeks, rats were sacrificed to collect urine, blood plasma, kidney tissue for detection of 24 h urinary protein, creatinine and renal pathological changes. Aldosterone concentration in plasma and kidney tissue was detected by radioimmunoassay; E-cadherin, α-SMA protein expression by immunohistochemistry, Western blotting; E-cadherin, α-SMA mRNA expression by RT-PCR. Results Compared with N group, serum creatinine, urinary protein excretion in the DN rats were significantly higher(P<0.01, respectively), E-cadherin protein and mRNA were significantly reduced (P<0.01, respectively), α-SMA protein and mRNA expression was up-regulated (P<0.01, respectively). Aldosterone level of kidney tissue in DN rats was increased obviously[(24.71±5.30) ng/g vs (16.38±2.85) ng/g, P<0.01], which was positively correlated with urinary protein excretion, serum creatinine and α-SMA protein(r=0.737, 0.574, 0.688, P<0.01, respectively), and negatively correlated with E-cadherin protein (r=－0.659, P<0.01). While no significant difference was found in serum aldosterone among three groups. Compared with DN rats, urinary protein excretion, serum creatinine were reduced (P<0.01, respectively), E-cadherin protein and mRNA were increased (P<0.01, respectively), α-SMA protein and mRNA expression were decreased (P<0.01, respectively) in SP group rats. Conclusions Local aldosterone involves in renal epithelial-mesenchymal transition in diabetic nephropathy rat. Spironolactone can block the effect of aldosterone and play a role in renal protection.

Objective To explore the protective effect and mechanism of antioxidant N-acetyl-L-cysteine (NAC) on the cytotoxicity induced by iohexol in HK-2 cells. Methods The incubated HK-2 cells were divided into four groups: control group, iohexol group, NAC group, and NAC+iohexol group(pre-incubated with NAC and then co-incubated with iohexol). The cell viability was tested by CCK-8 assay; cell apoptosis was determined by Hoechst 33342 fluorescence staining and flow cytometry with AnnexinⅤ-FITC/PI double staining. Intracelluar ROS was detected by flow cytometry with DCFH-DA fluorescence staining. The signaling transduction pathways were investigated by Western blotting and immunofluorescence staining. Results Iohexol decreased cell viability, and increased apoptosis in a dose- and time-dependent manner. In iohexol (100 gI/L, 6 h) group, ROS was increased by 1.30-fold of control (P<0.05). In NAC(5, 10, 15 mmol/L)+ iohexol groups, the cell viability was increased by 104%, 118%, 130% respectively, and iohexol group was 63% (P<0.05, respectively); apoptosis rate was decreased by 13.51%, 13.46%, 12.23% respectively, and iohexol group was 24.41% (P<0.05, respectively); ROS was decreased by 1.05-fold, 0.93-fold, 0.86-fold respectively, and iohexol group was 1.3-fold (P<0.05, respectively). Iohexol induced the increase of p53 phosphorylation and activity, then up-regulation of Bax and down-regulation of Bcl-2 protein expression. Iohexol induced the release of cytochrome C from mitochondria to cytoplasm, all of which caused final activation of caspase-3. The expression levels of p53, Bax and caspase-3 were decreased, while Bcl-2 protein expression level was increased by NAC. Conclusions Iohexol induces the increase of apoptosis rate and ROS generation in HK-2 cells. NAC attenuates this iohexol-induced cytotoxicity by decreasing intracelluar ROS, which is mainly through the intrinsic pathway.

Objective To explore the role of dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) in the tubulointerstitial lesions of immune-mediated nephrotoxic nephritis (NTN) and the intervention regulation by anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb). Methods WKY rats were randomly divided into control, NTN and PsL-EGFmAb-treated groups. The rats in NTN group were injected with 1 ml nephrotoxic rabbit serum per kilogram of rat body weight; the ones in PsL-EGFmAb-treated group were injected with 2 mg PsL-EGFmAb per kilogram of rat body weight simultaneously and 2 h later after nephrotoxic rabbit serum injection; and those in control group were injected with equal volume of 0.9% saline. Renal function and pathology were observed at day 4, 7 and 14 after the induction of NTN. Distribution of DC-SIGN+ dendritic cells(DCs) in renal tissues was measured by immunofluorescence. Real-time PCR was performed to examine the expression of P-selectin, RANTES, TNF-α, IL-10, IFN-γ and IL-4. Expression of MHCⅡ, CD80 and DC-SIGN on dendritic cells was analyzed by flow cytometry. Transendothelial migration was used to detect the ability of DCs migration. DCs ability to activate T cells was determined by mixed lymphocyte reaction (MLR). ELISA was used to detect the concentration of IFN-γ and IL-4 in the supernatant of MLR. Results At day 4, immature DC-SIGN+ DCs infiltrated the rat renal tubulointerstium of NTN group, matured at day 14, and enhanced the ability to migrate and activate T cells. The distribution of DC-SIGN+ DCs was significantly related to the form of crescent, tubulointerstial lesions and renal function. In addition, expression of chemokine RANTES and proinflammatory cytokine TNF-α continuously augmented since day 4, while anti-inflammatory cytokine IL-10 decreased after markedly increased at day 4. At day 14, IFN-γ/IL-4 mRNA increased, which was obviously related to DCs maturation. The intervention of PsL-EGFmAb supressed the expression of DC-SIGN and CD80 on DCs, depressed DCs maturation, migration and ability to activate T cells, down-regulated proinflammatory cytokines and up-regulated anti-inflammatory cytokines in kidney, and thus regulated Th1/Th2 bias. At the same time, kidneys showed the decrease of crescents, improvement of tubulointerstium damage and renal function. Conclusions DC-SIGN may mediate DCs tubulointerstitial infiltration. It may be also a potent regulator of local immune reaction imbalance and pathology of tubulointerstium. PsL-EGFmAb may depress DCs migration and down-regulate DCs maturation and function through DC-SIGN, and thus having a role in prevention and treatment.