Objective To elucidate the features of clinicopathology and mutation in Fabry disease complicated with thin basement membrane nephropathy (TBMN), and to investigate the kindred. Methods Data of clinicopathology and gene mutation of a female patient of Fabry disease complicated with TBMN admitted to the Department of Nephrology in our hospital were analyzed. Members of her kindred were investigated simultaneously. Results Proband was a 41-year-old Chinese woman who presented syndrome of Fabry disease and TBMN including angiokeratomas, chronic pain, tinnitus, vertigo, left ventricular hypertrophy and nephropathy as proteinuria, microscopic hematuria and hypertension. A percutaneous renal biopsy was performed on the proband, which was consistent with FSGS and vaculization of podocytes by light microscopy. Electron microscopy showed concentric lamellated inclusions in some podocytes. Diffuse thinning of glomerular basement membrane (GBM) with a mean thickness of (216±31) nm was found. The diagnosis of TBMN with suspected Fabry disease was identified. Family screening showed that her daughter had microscopic hematuria, tinnitus and neuropathic pain. One of her sisters only had microscopic hematuria. The activity of α-galacsidase A（α-Gal A）enzyme in the proband and her daughter was 33 units and 75 units respectively (the normal range is 100 to 500 units). They all carried the novel GLA mutation 1208 ins 21 bp and COL4A3 SNP c: 3627G>A(p:M1209I). While her sister who only had microscopic hematuria just carried the variant of COL4A3 gene—c: 3627G>A (p:M1209I), and had the normal activity of α-Gal A with no mutation of GLA. Conclusion TBMN should be considered in the patients of Fabry disease with the condition of benign familial hematuria.

Objective To investigate whether the combination of maintenance hemodialysis (MHD) with hemoperfusion(HP) can improve the clearance rate of middle- and large-molecule uremic toxins so as to improve the quality of life and reduce the mortality in MHD patients. Methods A prospective, randomized and controlled clinical trial was carried out. One hundred MHD patients were selected and then randomly divided into two groups after four weeks of run-in period. HD+HP group received MHD alone 2 times a week and the combined treatment of HD with HP (HD+HP) once a week, whereas HD group received MHD alone 3 times a week. The follow up lasted for mean 2 years. The primary outcome was the death of patients. Secondary end points included clinical data, leptin, high sensitive C-reactive protein (hsCRP), interleukin 6 (IL-6), β2 microglobulin (β2-MG), parathyroid hormone (PTH), tumor necrosis factor α(TNF-α) and the indexes of dimensions of Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36 Chinese Edition ). Results At the end of the two-year observation, the serum concentration of leptin, hsCRP, PTH, IL-6, β2-MG and TNF-α, systolic blood pressure(SBP), diastolic blood pressure(DBP), heart rate(HR), cardiothoracic ratio, left ventricular mass index (LVMI), EPO dose and the types of antihypertensive drugs used were lower in HD+HP group as compared to HD group (all P<0.05). HD+HP group had higher hemoglobin(Hb), ejection fraction (EF) and body mass index (BMI) (all P<0.05). No significant differences between two groups were found in terms of serum albumin (Alb), serum iron (SI), total iron binding capacity (TIBC), cardiac output (CO), Kt/V, early/atrial mitral inflow velocities (E/A) (all P>0.05). Besides, the SF-36 indicated that the total score of overall dimensions in HD+HP group was higher (P<0.05) and the quality of life of HD+HP group was evidently better as compared to HD group. The Kaplan-Meier survival curves for the 2-year observation period showed that patients in HD+HP group had obvious survival advantage, while Log-rank test results showed P<0.05. No serious adverse incidents occurred during the HD+HP treatment. Conclusion HD+HP is superior to HD in eliminating regularly middle- and large-molecules uremic toxins and has a potential role in improving the quality of life and survival rate of MHD patients.

Objective To investigate the effects of repeated low dose intravenous infusion of low molecular weight iron dextran and iron sucrose on oxidative stress in chronic renal failure(CRF) rats. Methods CRF model was established by 5/6 subtotal nephrectomy (5/6 Nx). Four weeks after removing the right kidney, successful rats were randomly divided into low molecular weight iron dextran group, sucrose iron group and CRF control group. The sham group was established simultaneously. The dose of iron administrated in each rat was similar in iron dextran group and sucrose iron group. There were 6 rats in each group. Animals were observed for 6 weeks，then the blood, urine and renal tissue samples were collected, and indexes of renal function, anemia, iron status and oxidative stress were investigated. Results The hemoglobulin (Hb) level in iron groups was significantly higher as compared to control group (P<0.05) but was not significantly different between two iron groups. The levels of serum iron, ferritin and saturation rate of transferring (TS) were obviously lower in control group as compared to sham group (P<0.05). Levels of above 3 indexes were significantly higher in two iron groups as compared to control group (P<0.05), but were not significantly different between two iron groups. Concentration of plasma advanced oxidation protein products (AOPP) was obviously higher in two iron groups than that in control group [(127.84±21.19) μmol/L, (134.21±29.38) μmol/L vs (81.83±19.93) μmol/L, P<0.05]. Plasma malonaldehyde (MDA) was significantly higher in iron sucrose group than that in iron dextran group [(6.06±0.73) nmol/L vs (4.99±0.80) nmol/L, P<0.05]. Serum levels of superoxide dismutase (SOD) and total anti-oxidant capacity (TAOC) had no significant differences among three CRF groups. Concentration of plasma glutathione peroxidase (GSH-Px) was significantly decreased in three CRF groups as compared to sham group (P<0.05), while plasma GSH-Px was significantly lower in sucrose iron group than that in iron dextran group and control group [(2123.11±74.78) nmol&#8226;ml-1&#8226;min-1 vs (2352.84±163.90) nmol&#8226;ml-1&#8226;min-1, (2310.23±125.99) nmol&#8226;ml-1&#8226;min-1, P<0.05]. Conclusions Injection of intravenous iron can partially improve the anemia and the iron status indexes in 5/6 Nx CRF rats. Repeated low dose intravenous infusion of iron dextran and iron sucrose can aggravate the oxidative stress state in CRF rats, and the iron sucrose is worst.

Objective To observe the formation of asymmetric dimethylarginine(ADMA) and the expression of dimethylarginine dimethylaminohydrolase 2 (DDAH-2) of human umbilical vein endothelial cells(HUVECs) stimulated by uric acid (UA), and to explore the role of ADMA-DDAH axis in the vascular endothelial dysfunction induced by uric acid. Methods HUVECs were cultured in M199 medium supplemented with 10% FBS. Cells were exposed to different concentrations of UA (0, 60, 120 mg/L) for 6 h and 24 h. Under different concentrations and times, the level of ADMA in cell suspension was detected by high performance liquid chromatography (HPLC) technique; the gene and protein expressions of DDAH-2 were detected by RT-PCR and Western blotting; the fluorescence intensity of intracellular 2’,7’-dichlorofluorescein (DCF) which represented the productions of ROS was detected by the flow cytometry (FCM). The activity of DDAH-2 in HUVCEs which were exposed to different concentrations of UA (0, 60, 120 mg/L) or UA (120 mg/L) +NAC (10 mmol/L) for 24 h was estimated by directly measuring the amount of ADMA metabolized by the enzyme and the role of NAC in the activity was studied. Results The expression of ADMA induced by urid acid was dose-depent and higher at 24 h than that at 6 h in the same dosage(all P<0.05). The dosage and stimulation time of UA did not have any influence on the expression of intracellular DDAH-2 (all P>0.05). When HUVECs exposed to UA(120 mg/L) for 24 h, the production of intracellular ROS was significantly increased while the activity of DDAH-2 was decreasesd (all P<0.05) as compared to 60 mg/L stimulation. This effect could be inhibited by the intervention of anti-oxidant NAC. Conclusions The high UA stimulation on HUVECs can increase the expression of intracellular ROS and inhibit the activity of DDAH-2 which increases the concentration of ADMA by decreasing the degradation of ADMA as well as the formation of NO. DDAH-ADMA axis may participate in the vascular endothelial dysfunction induced by UA.

Objective To explore the role of oxidative stress in the epithelial-to-mesenchymal transition (EMT) of tubular epithelial cells and the protective effect of probucol in rat model with diabetic nephropathy(DN). Methods Thirty SD rats were randomly divided into normal control group, DN group, probucol treatment group (supplemented 1% probucol dietary). Twenty-four hours urinary protein excretion (UTP) was measured at the 3rd, the 8th and the 12th week respectively. The biochemical indicators including blood glucose (BG), lipids [triglyceride (TG), total cholesterol (TC)], low-density lipoprotein (LDL), serum creatinine (Scr), creatinine clearance rate (Ccr),kidney tissue malondialdehyde (MDA) level and glutathione peroxidase (GSH-Px) activity were assessed at the end of the 12th week in all groups. The renal pathological changes were evaluated by hematoxylin & eosin(HE) and Masson staining. The protein expression of specificity protein 1 (Sp1), α-smooth muscle actin (α-SMA) and E-cadherin was also detected and analyzed by immunohistochemistry and Western blotting. Results Compared with the normal control group, the BG, TC, LDL, Scr, 24 h UTP and MDA level of renal tissue increased significantly and the Ccr reduced in the rats of DN group (all P<0.01). The pathological scores and the expression of Sp1 and α-SMA in renal tissue were higher in the DN animals than that in the other animals (all P<0.01), the expression of E-cadherin downregulated significantly in the DN animals (P<0.01). The MDA level of renal tissue was positively correlated to the expression of α-SMA and Sp1 protein in DN group (r＝0.896, P<0.01; r＝0.862, P<0.01, respectively), and negatively correlated to the expression of E-cadherin protein (r＝-0.673, P<0.01). In the diabetic animals treated with probucol, the Scr, 24 h UTP, pathological scores, MDA content,expression of Sp1 and α-SMA in renal tissue were lower than those in the diabetic animals (all P<0.01). The Ccr and the expression of E-cadherin upregulated obviously (all P<0.01). Conclusion Oxidative stress plays an important role in the EMT process of tubular epithelial cells. Probucol can slow down renal disease progression in DN rats through anti-oxidant, downregulating the expression of Sp1 protein and inhibiting the renal tubular EMT.

Objective To investigate the effect of intermedin(IMD) on tubular cells apoptosis induced by renal ischemia/reperfusion(I/R) injury and its associated mechanism. Methods A total of twenty-four male Wistar rats were randomly divided into four groups (control group, I/R group, empty plasmid group and IMD group). One week after the removal of right kidney, ultrasound plasmid was used to transfect empty or IMD plasmid into the left kidney. Renal I/R model was made by clasping the left renal artery for 45 minutes. Tubular cell apoptosis was determined by TUNEL. Expression of Bax, Bcl-2, Fas was detected by semi-quantitative RT-PCR. Activity of caspase-8 and caspase-9 was evaluated with commercially available kits respectively. Protein level of caspase-3 was measured by Western blotting analysis. Results Compared with control group, apoptosis of tubular epithelial cells, expression of Bax and Fas, activities of caspase-8 and caspase-9, as well as protein level of caspase-3 were all significantly increased in I/R group (all P<0.05). IMD pre-treatment significantly inhibited all these effects (all P<0.05). There were no differences of above parameters between empty plasmid group and I/R group. Conclusion IMD pre-treatment protects against renal I/R injury by inhibion of tubular epithelial cell apoptosis.

Objective To investigate the renoprotection of tubular L-FABP in murine IgA nephropathy (IgAN） induced by bone marrow transplantation(BMT). Methods IgAN models were reconstituted by BMT from IgAN-prone mice into mice (Tg) transgenically tubular overexpressing human L-FABP (hL-FABP) and wild type (WT) mice. These recipients were sacrificed at 6 and 12 weeks after BMT and their kidneys were collected. The expressions of hL-FABP, fibronectin (FN) and monocyte chemoattractant protein-1(MCP-1) mRNA were detected by real-time PCR. hL-FABP, FN, type Ⅳ collagen (Col Ⅳ), hemeoxygenase-1(HO-1) and 4-hydroxy-2-nonenal (4-HNE) modified proteins were detected by Western blotting. The distribution of hL-FABP and FN protein in kidney was detected by immunohistochemistry. The level of serum IgA, urinary albumin and urinary hL-FABP was detected by ELISA. Results (1) IgAN was reconstituted in both Tg and WT mice by BMT: mesangial IgA deposition and up-regulation of serum IgA. The levels were not significantly different between two groups (Tg-ddY and WT-ddY). (2) hL-FABP was expressed in proximal tubular cells of normal Tg mice. The mRNA (1.62±0.32 vs 0.46±0.09, P<0.01) and protein expression (1.74±0.76 vs 1.14±0.31, P<0.01) of hL-FABP was up-regulated in Tg-ddY kidney and urinary hL-FABP level (?滋g/g creatinine) was significantly increased (59.87±26.75 vs 31.01±14.86, P<0.05) at the 6th week after BMT. (3) WT-ddY mice showed a significantly higher urinary albumin level (mg/L) (828±656 vs 82±22, P<0.01), severer mesangial matrix expansion (P<0.01)，more glomerular FN and Col Ⅳ deposition at the 12th week. (4) Up-regulation of renal hL-FABP was associated with significant suppression of renal HO-1 expression (P<0.05), accumulation of 4-HNE modified proteins (P<0.05) and MCP-1 mRNA expression (P<0.01) in Tg-ddY mice. Conclusion Tubular L-FABP may lessen the progression of glomerular damage at early stages of IgAN by reducing oxidative stress and inflammatory mediators.

Objective To explore whether the change of S phase kinase-associated protein 2 (Skp2) expression could regulate mesangial cell proliferation. Methods Skp2 siRNA and control siRNA, pIRES-GFP-Skp2 plasmid and pIRES-GFP plasmid were designed and synthesized. Cell transfection was performed by Lipofectamine 2000. Skp2 mRNA and protein levels were detected by semiquantitative PCR and Western blotting respectively. Primary culture rat mesangial cells were divided into 6 groups: 0%FCS, 20%FCS, 10%FCS+pIRES-GFP plasmid, 10%FCS+pIRES-GFP-Skp2 plasmid, 20%FCS+control siRNA, 20%FCS+Skp2 siRNA. Cell number was detected by MTT. S phase entry was measured by BrdU labeling. Cell cycle profile was determined by flow cytometric analysis. Results Skp2 mRNA level was significantly down-regulated by Skp2 siRNA compared to control siRNA. Skp2 protein level increased after pIRES-GFP-Skp2 plasmid transfection compared to pIRES-GFP plasmid. MTT, BrdU labeling and cell cycle profile demonstrated that cell number (A: 0.419±0.088 vs 0.305±0.036, P＜0.01) and S-phase cells (BrdU labeling positive cell: 0.21±0.04 vs 0.15±0.03, P＜0.01；S-phase cell number：20.18±0.64 vs 14.33±0.37, P＜0.01) obviously increased after Skp2 plasmid transfection, while decreased after Skp2 siRNA transfection compared to control groups(A: 0.328±0.069 vs 0.482±0.133, P＜0.01; BrdU labeling positive cell: 0.17±0.01 vs 0.24±0.00, P＜0.01; S-phase cell number: 16.52±0.75 vs 23.81±1.25, P＜0.01). Conclusion Over-expression of Skp2 stimulates mesangial cell proliferation while down-regulated expression of Skp2 inhibits mesangial cell proliferation.

Objective To observe the change of Wnt-β-catenin signaling’s location and expression in kidney repair after acute kidney injury induced by ischemla reperfusion (I/R). Methods Ischemia reperfusion injury in BAT-gal reportor mice was made and blood sample was taken from tails on the 1th day after injury. Mice were sacrified on the 2th or 7th day and kidneys and blood were collected. Renal pathological change was observed by PAS stain. The changes of location and expression of Wnt-β-catenin signaling were detected by immunofluorescence costainning X-gal-LTL, X-gal-NKCC2, X-gal-DBA respectively. The protein expressions of the Wnt4 and co-receptor Lrp6 were assessed by Western blotting. Results PAS-stained kidney sections showed desquamative or flattened epithelia, necrotic debris on day 2 and regenerating tubules on day 7. An injury-induced enhancement of the Wnt pathway response (X-gal staining) in kidney cortex and out-medulla. Immunolabelling of kidney sections from injured BAT-gal mice revealed that X-gal staining was detected in kidney epithelial cells (double-labelled with LTL or NKCC2). Western blotting showed the Wnt4 protein was up-regulated and phospho-Lrp6, indicative of active canonical Wnt signaling, was noted in kidney cortex from day 2 after I/R, but in control kidney cortex pLrp6 was not detected. Conclusion Wnt-β-catenin signaling is activited after acute kidney I/R injury and is required for tubular epithelial repair and regeneration following kidney I/R injury.

Objective To investigate the clinic features in long-term peritoneal dialysis (PD) patients with the first episode of dialysis-related peritonitis and the risk factors. Methods In this retrospective study, 315 PD patients who experienced the first episode of peritonitis from January 2000 to December 2009 were recruited. All the patients were divided into two groups according to the duration of PD treatment: group A (<36 months, n=261) and group B (≥36 months, n=54). Clinical information including demographic character, primary renal disease, biochemical laboratory data, etiology data, treatment methods and clinical outcomes was collected. Results Among 315 patients, 61.0% was male and the mean age was (55.7±15.9) years. The primary renal diseases were glomerulonephritis (54.6%), and then diabetic nephropathy (20.6%). The median duration of PD was 8.4 months in group A and 49.4 months in group B. No significant difference was found in serum potassium and serum albumin levels between the two groups, but the levels of both indexes were below normal range. The most common cause of peritonitis was connection contamination in both groups (48.2% vs 45.2%). There were no significant differences in the incidence of gram-positive and gram-negative peritonitis between the two groups, but group B had higher drug resistance rate in gram-positive peritonitis than that in group A (46.2% vs 19.1%, P=0.035). The incidence of fungus infection in group B was significantly higher than that in group A (17.8% vs 6.4%, P=0.011). Treatment response was lower in group B than that in group A (P=0.000) and the clinical outcome in group B was worse than that in group A (P=0.000). Compared to gram-positive peritonitis, culture-negative peritonitis and peritonitis without culture, the incidences of treatment failure in group B were higher than those in group A (23.1% vs 1.5%, 46.2% vs 6.7%, 22.2% vs 0%, all P<0.05). By Logistic regression analysis, fungus infection, longer duration of PD treatment and lower serum albumin level were risk factors for the treatment failure of PD-related peritonitis (P=0.000, 0.002, 0.025). Conclusions The clinical outcome of the first episode of peritonitis in long-term PD patients is poorer than that in short-term PD patients. Fungus and drug-resistant bacteria infection, as well as malnutrition are the risk factors for the treatment failure of PD-related peritonitis in long-term PD patients.

Objective To investigate the clinicopathological characteristics of non-diabetic renal diseases (NDRD) in the patients with diabetes mellitus． Methods Clinicopatholigical data of 202 patients with diabetes mellitus and NDRD identified by renal biopsy from January 1st, 2003 to December 31st, 2010 were analyzed retrospectively. All the patients were divided into three groups: the young (≤35 years old), the middle-aged (36-59 years old) and the elder (≥60 years old). Clinicopathological characteristics were compared among 3 groups. Results In the young group (n=33), 42.4% of patients presented as chronic glomerulonephritic syndrome, while 36.4% as IgA nephropathy for pathology. In the middle-aged group (n=136), 35.3% of patients presented as chronic glomerulonephritic syndrome, 27.2% as nephritic syndrome, 17.6% as chronic renal failure, 14.7% as latent glomerulonephritis, and 5.1% as acute renal failure, while 42.6% as IgA nephropathy for pathology. In the elder group (n=33), 30.3% of patients presented as nephritic syndrome，30.3% as chronic renal failure, while 27.3% as membranous nephropathy for pathology. Conclusions In clinical manifestation, young patients are mainly chronic glomerulonephritic syndrome, middle-aged patients are diversified, and elder patients are mainly nephritic syndrome and chronic renal failure. In pathology, young and middle-aged patients are mainly IgA nephropathy, and elder patients are mainly membranous nephropathy.

Objective To investigate the inflammatory response and apoptosis of peripheral blood mononuclear cells (PBMCs) and their regulation by triptolide (TP) in IgA nephropathy (IgAN) patients. Methods Blood samples were collected from 29 IgAN patients and 16 healthy individuals. TNF-α and IL-6 concentrations were measured by ELISA and NO concentration by Griess reagent in the plasma of samples. PBMCs were isolated from IgAN patients and cultured in vitro, and subsequently activated by PHA (10 mg/L). The cytotoxicity of different TP concentrations was assayed by MTT and two non-toxic concentrations (12.5 μg/L or 25.0 μg/L) were selected for treatment. TNF-α, IL-6 and NO concentrations were measured in the culture media collected from PBMCs cultures activated by PHA (10 mg/L) and treated with TP (12.5 μg/L or 25.0 μg/L). The PHA-activated, TP-treated cells apoptotic rate was analyzed by FACS using Annexin V-FITC staining. The expression of Bcl-2, Bax, caspase-9 and caspase-3 were detected by RT-PCR and Western blotting from lyses of PHA-activated with or without TP-treated cells. Results The serum concentrations of TNF-α [(131.57±50.61) ng/L vs (30.24±18.93) ng/L, P<0.01], IL-6[(76.36±25.21) ng/L vs(35.08±16.59) ng/L, P<0.01] and NO[(46.36±12.93) μmol/L vs(26.61±10.87) μmol/L, P<0.01] were significantly increased in IgAN patients compared to healthy individuals. PBMCs viability in culture decreased after TP treatment in a dose-dependent manner. TP also inhibited TNF-α, IL-6 and NO levels in the media of PHA-activated PBMCs in culture and induced PBMCs apoptosis. The expression of Bcl-2 decreased markedly and Bax, caspase-9 and caspase-3 increased significantly after TP treatment（all P<0.05). Conclusions The PBMCs from IgAN patients are in a highly activated state, and have a high apoptotic rate. TP treatment induces benificial effects in IgAN patients by inhibiting the activation of PBMCs by activating pro-apoptotic pathway.

Objective To clarify the clinicopathological features of renal amyloidosis in order to achieve early diagnosis and treatment. Methods Clinicopathological data of 26 biopsy-proven renal amyloidosis cases in Department of Nephrology, Zhongshan Hospital, Fudan University between 2006 and 2010 were analyzed retrospectively. Immunohistochemistry and immunofluorescence of amyloid A protein, immunoglobulin light chains such as ?资、?姿 were performed on renal specimens for further classification. Results Age of 26 patients ranged from 40 to 77 years old, average (58.54±10.07) years. Twenty-two out of 26 patients (84.62%) were treated in local hospital before admitted to our department, and 21 patients (95.45%) were misdiagnosed as chronic primary glomerulonephritis. The prominent clinical manifestations of renal amyloidosis were nephrotic syndrome (17 cases, 65.38%), decreased blood pressure (16 cases, 61.53%), organ enlargement (8 cases, 30.77%) and bodyweight loss (6 cases, 23.08%). Fourteen out of 25 patients (56.00%) were found to have monoclonal light chains in serum by immunofixation electrophoresis. Three patients with mild pathological changes who had no confirmable Congo red stain were confirmed by electron microscopy. Twenty-three (88.46%) patients were diagnosed as AL amyloidosis, one (3.85%) as AA amyloidosis, one was strongly suspected of hereditary amyloidosis, and one was undetermined. Conclusions Renal amyloidosis is frequently misdiagnosed. Middle-aged and old nephrotic patients with decreased blood presure, organ enlargement and bodyweight loss may be the most helpful clues of the disease. Most patients have monoclonal light chains in serum or urine. Renal biopsy, especially electronic microscopy plays a crucial role in the early diagnosis of renal amyloidosis. Immunohistochemistry is important for patients with renal amyloidosis in pathological classification and treatment.

Objective To elucidate the association between aquaporins (AQPs) expression in kidney tissue and edema of nephrotic syndrome (NS) patients. Methods NS patients were divided into edema group (14 cases) and non-edema group (8 cases). Ten patients without NS were used as control group. Expressions of AQP1, AQP2, and AQP4 in renal tissues of 3 groups were detected by immunohistochemistry with standard techniques and semi-quantitative analysis. Association between AQPs expression and edema was examined. Results The positive index of AQP1 expression in proximal tubules in edema group was 0.0373±0.0110, which was significantly lower as compared to non-edema group (0.0510±0.0120) and control group 0.0574±0.0100), while the difference between non-edema and control groups was not significant. The positive index of AQP1 expression in glomerulus was 0.0106±0.0037 in edema group, which was significantly higher than that in non-edema group (0.0021±0.0013) and control group (0.0020±0.0012), while no significant difference was found between the last two groups. AQP2 mainly localized in the collecting duct system. The positive indexes of AQP2 expression were 0.0498±0.0081, 0.0370±0.0072 and 0.0255±0.0103 in edema group, non-edema group and control group, respectively. The differences were significant among 3 groups. AQP4 expression was not found in the renal cortex and collecting duct system. Conclusions AQPs expression is different in renal tissues of NS patients. AQP2 may play an important role in the edema of NS patients, and AQP1 may involve in the occurrence of edema.

Objective By reporting a fatal case of severe lupus nephritis (LN) complicated with invasive aspergillus pneumonia and meningitis and reviewing the associated literatures, to provide a way of early diagnosis and proper management for the patients suffering from systemic lupus erythematosus (SLE) and invasive fungus infection (IFI). Methods The onset, diagnosis and treatment course of the disease were described and associated literatures were reviewed to analyze and summary the diagnostic methods, common pathogenic bacteria and predisposing factors of SLE patients with IFI. Results Application of IFI guideline for cancer patients and those undergoing hematopoietic stem cell transplantation could be helpful in the early diagnosis and treatment of SLE patients complicated with IFI. The most common pathogen of SLE patient suffering from IFI was cryptococcus neoformans and aspergillus, not candida albicans. The main predisposing factors were high lupus activity and immunosuppressant. Conclusions Guideline of IFI for cancer patients and those undergoing hematopoietic stem cell transplantation is also helpful for the SLE patients complicated with IFI. The most common pathogens of IFI in SLE patients are cryptococcus neoformans and aspergillus. The predisposing factors are high lupus activity and immunosuppressant.

Objective To evaluate the effects of AngⅡ on apoptosis of podocytes and explore the signaling pathway of nephrin in preventing AngⅡ-induced podocyte apoptosis. Methods Differentiated mouse podocytes were exposed to AngⅡat different concentrations for 18 h or at 10-8 mol/L for variable incubation times. Undifferentiated mouse podocytes were transfected using lipofectamine 2000 with the pcDNA3.1-mNPHS1 plasmid and stably transfected cell lines were generated with G418 selection. In separated experiments, untransfected mouse podocytes (MPC) and stably transfected podocytes with pcDNA3.1-neo and pcDNA3.1-mNPHS1 were exposed to AngⅡ(10-8 mol/L) or LY294002 (a selective Akt inhibitor, 50 μmol/L) for indicated times. Apoptosis was evaluated by flow cytometry. The expression of nephrin was assessed by quantitative real-time PCR，immunofluorescence and Western blotting. The phosphorylation level of Akt was determined by Western blotting. Results (1) AngⅡ promoted podocyte apoptosis in a dose-and time-dependent manner. Pretreatment with losartan significantly prevented Ang Ⅱ-induced apoptosis. (2) Nephrin mRNA and protein were obviously decreased in podocytes exposed to 10-8 mol/L AngⅡ for at least 12 h than those in vehicle-treated cells (P<0.05). (3) AngⅡexposure for more than 15 min inhibited the phosphorylation of AKT in MPC, which was dramatically reversed by pcDNA3.1-mNPHS1 transfection, but not by pcDNA3.1-neo transfection. (4) Podocyte apoptosis was promoted by LY294002. Conversely, AngⅡ-induced podocyte apoptosis was significantly alleviated by pcDNA3.1-mNPHS1 transfection. Conclusion AngⅡ induces mouse podocyte apoptosis which is suppressed by overexpression of nephrin through PI3K-Akt signaling pathway.

Objective To investigate the influence of TGF-β receptor subtypes expression and their downstream signaling Smad proteins on rat renal interstitial fibrosis induced by unilateral ureteral obstruction (UUO). Methods A total of 90 rats were randomly divided into three groups: normal control (CON), sham operation(SOR) and UUO group, and sacrificed 1, 3, 7, 14 and 21 days after operation. Serum creatinine and urea nitrogen were detected to assess renal function. PAS and Masson staining were performed to observe histological damage in the kidneys. Quantitative RT-PCR was used to define expression of mRNA encoding TGF-β receptor subtypes and their downstream signaling Smad proteins in kidney tubular cells. Real-time PCR, Western blotting and immunofluorescence were used to monitor the time-related expression of the TGF-β receptor subtypes and their downstream signaling Smad proteins in kidney. Results Compared with the CON group, serum creatinine and urea nitrogen in UUO groups increased at day 3 after operation (P<0.05) and reached their peak 21 days after operation (P<0.01). Obvious inflammatory cell infiltration was observed in UUO group 3 days after operation, while renal tubular atrophy and renal interstitial fibrosis were observed in UUO group 14 days after operation. The mRNA expressions of ALK-5, ALK-7 and TGF-βRⅡincreased significantly in UUO group 3 days after operation(all P<0.05) and reached their peaks 14 days after operation(all P<0.01). The mRNA expression of ALK-6 decreased significantly in UUO group 3 days after operation (P<0.05) and reached its lowest level 14 days after operation (P<0.01). The changes in the protein level of those receptors were consistent with their mRNA expressions. The protein expressions of Smad2/3 and p-Smad2/3 increased significantly in UUO group at day 3 (all P<0.05) and reached their peak at day 14 after operation (all P<0.01). Conclusion Expressions of TGF-β receptor subtypes ALK-5, ALK-6, ALK-7, TGF-βRⅡand their downstream signaling Smad2 and Smad3 proteins may influence the progress of renal interstitial fibrosis, tubular atrophy and inflammatory cell infiltration in UUO model rats.

Objective To explore the role of ERK1/2 in the expression of the type-1 plasminogen activator inhibitor (PAI-1) induced by parathormone(PTH) in human renal tubular epithelial cell line HK-2 cells. Methods Various concentrentions of PTH and manifold durations were applied in the test. The expression of PAI-1 mRNA and protein in HK-2 cells was measured by RT-PCR and Western blotting, respectively. Besides, ERK1/2 protein was detected by Western blotting before the ERK1/2 inhibitor incubated with the HK-2 cells or after. Results The expression of PAI-1mRNA and protein was gradually up-regulatad along with the increasing concentrations of PTH (10-12-10-10 mol/L). The maximum level of PAI-1 mRNA and protein was detected in 10-10 mol/L PTH and was 4.01 and 3.81 times of control group. Otherwise, the decreased expression of PAI-1 was found while the concentrations of PTH were beyond 10-10 mol/L. The levels of PAI-1 mRNA and protein were increased in pace with time from 12 to 72 hour, in time-dependent manner, which was 4.06 (12 h) and 4.03 (72 h) times of 0 hour group. The levels of ERK1/2 and PAI-1 were ascended after 10-10 mol/L PTH incubated with the HK-2 cells (all P<0.01). Howerver, both of them decended after cells were pretreated by the ERK1/2 inhibitor (all P<0.01), but were still higher than those of control group (all P<0.05). Conclusion ERK1/2 kinase system partly participates in the regulation of PAI-1 induced by PTH in HK-2 cells.

Objective To investigate the effect of FUT8-siRNA on transforming growth factor β(TGF-β)-Smad2/3 signalling pathway in renal tubular epithelial cells. Methods HK-2 cells were divided into six groups: normal group, negative control group, TGF-β1 group, TGF-β1 with FUT8 interference group, TGF-β1 with negative control group, FUT8 interference group. RNAi was performed to silence the expression of FUT8 gene, then immunofluorescent analysis was used to detect the expression of core fucose in the HK-2, immunoprecipitation and lectin blotting were performed to detect the core fucosylation of TGF-βRⅡ and ALK-5, and detect the change of Smad2/3 and p-Smad2/3 in HK-2 cells after FUT8 gene was silenced. Results Compared with the normal and negative control group, incubation with 5 μg/L TGF-β1 for 48 h could significantly up-regulate the core fucosylation of HK-2 cells, enhance the protein expression of TGF-βRⅡ and ALK-5 (P<0.05), markedly increase the expression level of p-Smad 2/3 (P<0.05) and cause it to nuclear translocation in HK-2 cells. While FUT8siRNA could inhibit the above up-regulation of TGF-βRⅡ and ALK-5 (P<0.05), suppress the increase of p-Smad 2/3 (P<0.05) and its nuclear translocation without disturbing the protein expression of TGF-βRⅡ and ALK-5. Conclusion FUT8-catalized core fucosylation of TGF-βRⅡ and ALK-5 is needed to fulfill their functions, and blocking core fucosylation of TGF-βRⅡ and ALK-5 leads to the inhibition of TGF-β-Smad2/3 signalling pathway in HK-2 cells.

Objective To investigate the effect of high volume hemofiltration(HVHF) combined with mechanical ventilation(MV) on seawater respiratory distress syndrome(SW-RDS) canine models. Methods Ten nomal hybrid dogs were randomly assigned into two groups: MV group (MV group, n=5), all the animals only received MV after establishing model successfully; HVHF combined with MV group(HVHF+MV group, n=5), all were received HVHF plus MV after establishing model successfully. Both groups were observed for 4 hours. Mean arterial pressure (MAP), heart rate(HR), central venous pressure (CVP), arterial blood gas and venous plasma osmotic pressure were detected at baseline, 0 min (model establishment), 60 min, 120 min, 180 min, 240 min after treatment. Venous blood was collected to detect inflammatory mediators (IL-8, IL-6, TNF-α) at baseline, 0 min, 120 min, 240 min after treatment. The lung pathology was examined at the end of the experiment. Results (1)All the animals were suvival after four hous of treatment in both groups. (2)Partial pressure of oxygen (PaO2) and O2 saturation (SaO2) rised after four hours of treatment in both groups (P<0.05), and HVHF+MV group was better than MV group. After 4 hours of treatment, pH, actual bicarbonate(AB), bases excess (BE) in HVHF+MV group were significantly better than those in MV group (P<0.05), recovering to the baseline values. (3)MAP, HR, CVP were stable during the four hours of treatment, and compared with 0 min, there was no significant differences after 4 hours of treatment in both groups. There were no significant differernces at the same time of treatment in both groups. (4)Plasma osmotic pressure were stable during the four hours of treatment, and compared with 0 min, there was no significant difference in MV group. But in HVHF+MV group, osmotic pressure was significantly higher after 4 hours of treatment than that at the same time in MV group (P<0.05), and compared with 0 min and 180 min, those were higher too (P<0.01). (5)Compared with those at the same time in MV group, plasma inflammatory mediators(IL-8, IL-6, TNF-α) were significantly decreased after 4 hours of treatment in HVHF+MV group (P<0.01). After 4 hours of treatment IL-8, TNF-α in MV group were higher than those at the same time in 0 min (P<0.05). (6)Compared with those in MV group, there were less infiltration of neutrophils, edema and injury of alveolar epithelium from pulmonary pathology. Conclusions HVHF combined with MV can significantly improve hypoxemia and correct acidosis of SW-RDS model in canines. HVHF can effectively clear plasma inflammatory mediators and redundant water, improve pulmonary pathology changes. HVHF has no impact on MAP, HR and CVP of SW-RDS.

Objective To make a new IgA nephropathy (IgAN) animal model by infecting mice with Mycoplasma penetrans (Mpe). Methods Mice were infected through urinary tract with Mpe, Sp-4 medium or PBS, and combined with tail vein challenge to make experimental IgAN animal model. Pathologic changes were compared between new IgAN model and classical IgAN model. Results A new IgAN animal model was established successfully and the successful rate was 100%. There was no significant difference in IgA deposition rate or fluorescence intensity between our new IgAN animal model and the classical IgAN animal model.There was also IgG deposition found in 66.67% Mpe-infected mice. Light microscopy examination revealed compared with PBS or SP-4 medium control group, the proliferation of mesangial cells and the deposition of matrix were more serious in Mpe-infection group (all P<0.05), which was not signifcantly different with classical IgAN group. Besides, there was mononuclear cell infiltration in tubular interstitium and parietal cell proliferation in Bowman's capsule in Mpe-infection group. Conclusion By infecting mice with Mpe through urinary tract, IgAN animal model can be successfully made which may offer a new direction for IgAN pathogen research.