Objective    To evaluate the copy number variation of FCGR3B gene in Henan Han systemic lupus erythematosus (SLE) patients and healthy controls, and explore the association between FCGR3B gene copy number variants (CNVs) and lupus nephritis (LN) susceptibility in Henan  Han population.    Methods    FCGR3B CNVs was investigated in 142 SLE patients with nephritis, 187 SLE patients without nephritis and 328 healthy controls. A modified methodology based on competitive PCR named Multiplex AccuCopy^TM Kit was used to detect FCGR3B copy number. Clinical and laboratory data were collected retrospectively from the medical record. Logistic regression analysis was used to determine the association of FCGR3B copy number variants with LN susceptibility. Rank correlation was used to determine the correlations between FCGE3B copy number variants and clinical phenotypes of LN.    Results    No significant difference was detected in the copy number variations of FCGR3B in different groups. Low copy number of FCGR3B was more commonly seen in patients with nephritis (P=0.042), and was a risk factor for LN (OR=2.059; 95% CI: 1.081-3.921; P=0.028). However, high copy number (＞2) had no effect on SLE patients without nephritis(OR=1.152; 95%CI: 0.711-1.866; P=0.565) and LN patients (OR=0.838; 95%CI: 0.529-1.329; P=0.454). There were no associations between FCGR3B copy number variants and clinical phenotypes and immunologic characteristics of LN.    Conclusion    The low copy number of FCGR3B is a risk factor for LN in  Henan Han population.

Objective    To investigate the relationship between urinary sediments and renal pathological patterns in IgA nephropathy.    Methods    A total of 516 specimens of fresh fasting morning urine were collected. Urinary specimens were processed routinely and examined with phase-contrast microscopy. Urinary sediments were classified into three types according to the components: type Ⅰ: hematuria containing dysmorphic red blood cells(RBCs); type Ⅱ: active urinary sediments containing hematuria and casts of RBCs, and/or a few casts containing white blood cells or epithelial cells; type Ⅲ: advanced urinary sediments containing hematuria, and various casts consisted of red and white blood cells, renal tubular epithelial cells, and waxy casts. Pathological patterns of IgA nephropathy were classified according to the descriptive diagnosis of pathological lesions. Statistical analysis were performed using kappa test, Χ2 test, and significance was accepted at P＜0.05.    Results    The pathological patterns of 516 cases of IgA nephropathy included focal proliferative pattern in 75.2%, mesangial proliferative pattern in 7%, proliferative and sclerosing pattern in 6.8%, endocapillary proliferative pattern in 2.9%, crescentic pattern in 2.3% respectively. Hematuria was present in 74.8% (386/516 cases) of patients before renal biopsy, 15.7% of them had gross hematuria. Urinary sediments of 386 cases consisted of 54% of type Ⅰ, 35% of type Ⅱ, and 11% of type Ⅲ respectively. Type Ⅰ urinary sediment was present in focal proliferative pattern without active lesions, its concordance was 79.4%; while type Ⅱ were mainly seen in cases with glomerular and/or tubulointerstitial active lesions; type Ⅲ were seen in crescentic or endocapillary proliferative patterns and/or cases with active tubulointerstitial lesions. The concordance of type Ⅱ and type Ⅲ urinary sediments with active glomerular and tubulointerstitial lesions in IgA nephropathy was 79.2%. There was a significant difference in urinary sediment types between active and non-active pathological lesions (P＜0.01). Multivariate analysis indicated urinary sediment types independently predicted the active and non-active lesions in IgA nephropathy (P＜0.01, OR=7.268).    Conclusions    Urinary sediment analysis is an easy and valuable method to predict the active and non-active lesions of renal pathology in IgA nephropathy, can be used as a non- invasive indicator of clinical course.

Objective    To compare the clinical efficacy and side-effects of high dosage methylprednisolone combined with plasmapheresis or only with high dosage methylprednisolone treatment in patients with anti-neutrophil cytoplasmic antibodies (ANCA) associated vasculitis (AAV).   Methods    Forty-seven patients with clinicopathologically diagnosed as ANCA-associated vasculitis kidney damage, whose clinical manifestations were also consistent with rapidly progressive glomerulonephritis syndrome, were treated with high dosage methylprednisolone and plasmapheresis (PE group, n=22) or only with high dosage methylprednisolone (MP group, n=25). Patients received these two regimes were comparable in general conditions and clinicopathological parameters for the activity and severity of renal damage.    Results    At the 6 month, the serum level of ANCA were both lower significantly than before (P＜0.05), and the PE group had the advantage to the MP group (P=0.028). The renal function was improved both in the two groups, and the serum creatinine of PE group was significantly lower than that in the MP group (P=0.043). The BVAS scores were both significantly decreased in the two groups (P＜0.05), and the scores were lower in the PE group than that in the MP group (P＜0.05). Both the serum albumin and hemoglobin levels in the two groups were significantly increased after treatment (P＜0.05), while the differences before and after treatment  of the two levels between the two groups both had no significant difference (P＞0.05). Three patients developed to end stage renal failure in each group during the follow-up period. The incidences of gastrointestinal symptoms (12.0％ and 9.1％ in MP and PE group, respectively) and infectious complications (28.0％ and 22.7％ in MP and PE group, respectively) were similar between two groups.    Conclusions    Both the high dosage methylprednisolone combined with plasmapheresis treatment and the high dosage methylprednisolone treatment have significant effects on patients with ANCA associated vasculitis, while the high dosage methylprednisolone combined with plasmapheresis treatment may be more effective in decreasing serum level of ANCA and improving renal function .

Objective    To clarify the long-term renal prognosis and related risk factors of progression for IgA nephropathy (IgAN) patients who achieved remission under current therapy. To identify the target value of the serum albumin level for Chinese patients with IgAN.    Methods    The patients with biopsy-proven primary IgAN in Nephrology Department of Renji Hospital in Shanghai were studied．The survival of renal and the relationships between clinical parameters and renal outcome were assessed.    Results    A total of 369 patients between Jan 2005 and Dec 2010 were included with a median follow-up time of 49.0 (38.0-65.8) months. All the subjects had achieved a complete remission (CR) or partial remission (PR) following six months’ therapy after diagnosis. Progressive renal disease had occurred in 61 cases at the end of follow-up. Three variables had a significant independent effect on renal outcome in patients achieving remission under current therapy regimen for IgAN, including time-average serum creatinine (TA-Scr) [HR(95%CI): 1.03(1.01-1.04)], time-average serum albumin (TA-Alb) [HR(95%CI): 0.83 (0.69-0.99)], and eGFR ratio within one year [HR(95%CI): 0.00(0.00-0.01)]. By multivariate Cox analyses, each 1 g/L drop of TA-Alb was related with 17.2% increase in the risk of renal progression. The ROC curve indicated that combination of serum albumin at baseline and during a long-term had a more significant value in prediction of renal outcome than independent predictor alone. By Kaplan-Meier analyses, patients with TA-Alb﹤38 g/L had a 10.4 fold sincreased risk of progressive disease compared with that of TA-Alb﹥38 g/L.    Conclusions    IgAN patients with lower eGFR ratio, higher TA-Scr and lower TA-Alb would progress to ESRD more quickly, and serum albumin during follow-up is important for predicting IgAN progression.

Objective    To evaluate the effects on the mineral bone disorder using different calcium concentration citrate-based dialysate in maintenance hemodialysis (MHD) patients. To compare the concentrations of intact parathyroid hormone(PTH) with biointact PTH(1-84) in these patients.    Methods    Citrate dialysate with different calcium concentration (DCa l.75，DCa l.5，DCa l.25 mmol/L)were used in turn in 15 stable MHD patients each week．Serum tCa and iCa were measured by automatic biochemistry analyzer. The concentrations of iPTH and bio-iPTH were compared.    Results    (1) The patients treated with DCa l.75 citrate dialysate had increased serum iCa and tCa after dialysis, and PTH did not change significantly as compared to those findings before the dialysis. With the DCa l.5 citrate dialysate, serum iCa and tCa were kept stable and PTH level was increased. With DCa l.25 citrate dialysate, serum iCa and tCa decreased significantly and PTH decreased. (2)iPTH and bioPTH had excellent correlations. Variation of bio-iPTH was more correlated with the changes of calcium than iPTH.    Conclusions    Serum levels of iPTH, tCa and iCa can be kept stable in MHD patients treated with DCa 1.75～l.5 citrate dialysate. Bio-iPTH is a more sensitive marker for mineral bone disease than iPTH.

Objective    To observe the effect of high volume hemofiltration (HVHF) on the expression of CCAAT enhancer binding protein(CHOP) during the treatment of multiple organ dysfunction syndrome (MODS). To investigate the role of CHOP protein act in apoptosis pathway mediated by the endoplasmic reticulum stress.    Methods    Twelve Beagle dogs were subjected to hemorrhagic shock plus resuscltation and endotixemia to establish MODS model, then they were randomly divided into two groups: HVHF group (n=6) and MODS group (n=6). After endotoxin injection completed, the HVHF group received HVHF treatment for 24 hours; MODS group did not receive. Vivo experiments: Blood samples were obtained at different time points(before operation, 0 h, 6 h, 12 h, 24 h after the injection of endotoxin). The dogs were killed and the tissue samples from lung, liver and kidney were took, then the expression of CHOP mRNA was determined. Vitro experiments: human umbilical vein endothelial cells (HUVECs) were induced by two groups’ blood samples to establish the apoptosis model. Gene expression, protein quantification and cell apoptosis rate were determined before and after the interference.    Results    Vivo experiments: The levels of CHOP mRNA from lung, liver and kidney had no significant difference between the two groups (P＞0.05). Vitro experiments: (1)The expression of CHOP mRNA: Compared with MODS group, the expression levels of CHOP mRNA were significantly decreased in HVHF group at 6 h, 24 h after the injection of endotoxin (P＜0.05). Compared with before, the expression levels of CHOP mRNA in the two groups were both significantly decreased after CHOP siRNA interference (P＜0.05). (2)The expression of CHOP protein: Compared with MODS group, the expression levels of CHOP protein were significantly decreased in HVHF group at each time points (P＜0.05). Compared with before, the expression levels of CHOP protein in the two groups were both significantly decreased after CHOP siRNA interference(P＜0.05). (3)Endothelial cell apoptosis rate: Compared with the preoperative rate, the two group’s endothelial cell apoptosis rate was decreased significantly at each time points(P＜0.05). Compared with MODS group, the endothelial cell apoptosis rate was significantly decreased in HVHF group at each time points(P＜0.05). Compared with before, the endothelial cell apoptosis rate in the two groups was both significantly decreased after CHOP siRNA interference(P＜0.05).    Conclusion    In the treatment of MODS process, HVHF can reduce endothelial cell apoptosis which may be related to the inhibition of CHOP mRNA expression and protein synthesis.

Objective    To explore the effect of vitamin K2 on β-glycerophosphate(β-GP)-induced rat vascular smooth muscle cells (VSMCs) calcification and and the mechanism.    Methods    VSMCs were obtained from rat aortic, and identified by immunocytochemistry, then randomly divided into control group, high phosphorus group, vitamin K2 group (the group was settled three subgroups according to the concentration of vitamin K2 based on the high phosphorus medium, namely 10 μmol/L, 25 μmol/L, 50 μmol/L) and noggin (bone morphogenetic protein pathway inhibitor) group. Calcification was visualized by Alizarin red staining, calcium load in cells was quantified by o-cresolphthalein complexone method and alkaline phosphatase (ALP) activity was measured after stimulating 14 days, gene expressions of bone morphogenetic protein - 2 (BMP-2), SMAD1, SMAD7 and Runx2 mRNA were detected by RT-PCR, Runx2 protein levels was detected by Western blotting after stimulating 3 days.    Results    Compared with the cells in control group, high phosphorus induced cell calcification, increased ALP activity, up-regulated the expression of BMP-2, SMAD1, Runx2 mRNA (P＜0.05) and down-regulated the expression of SMAD7 (P＜0.01)，while compared with high phosphorus group, the calcium deposition, ALP activity and the expression of BMP-2, SMAD1, Runx2 mRNA were remarkably reduced in a dose-dependent manner by treatment with vitamin K2 (P＜0.05) and the expression of SMAD7 was increased (P＜0.01). Compared with high phosphorus group, SMAD1 and Runx2 expression in noggin group were remarkably reduced(P＜0.01).     Conclusion    Vitamin K2 inhibits β-glycerophosphate-induced VSMCs calcification which correlates with the suppression of the expression of osteoblast markers through the down-regulation of bone morphogenetic protein pathway.

Objective    To investigate the effect of rosiglitazone(RGZ) on peritoneal morphology, function and the expressions of Aquaporin 1 (AQP-1), vascular endothelial growth factor A(VEGF-A) and cyclooxygenase 2(COX-2) in uremic rat of peritoneal dialysis.    Methods    Thirty Sprague-Dawley rats were randomly divided into five groups. Group S (n=6) was subjected to sham operation. Group N (n=6) was subjected to nephrectomy with silicon catheter inserted, but no peritoneal exposure. Group P (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter, using 4.25% peritoneal dialysis fluid 10 ml twice a day for 2 weeks. Group R (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter, using 4.25% peritoneal dialysis fluid containing rosiglitazone (0.2 mg/kg) 10 ml twice a day for 2 weeks. Group GW (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter, using 4.25% peritoneal dialysis fluid containing rosiglitazone (0.2 mg/kg) and GW9662 (0.2 mg/kg) 10 ml twice a day for 2 weeks. After two weeks of dialysis, a 90 min peritoneal equilibration test was performed and the amount of ultrafiltration was accurately measured. The partial peritoneum tissues of rats were harvested and stained by hematoxylin-eosin (HE), then morphology changes of partial peritoneum were examined by light microscopy. The expression of AQP-1,VEGF-A and COX-2 in omentum were detected with immunohistochemistry assay. AQP-1, VEGF-A and COX-2 mRNA were detected by qRT-PCR.    Results    Morphology changes of partial peritoneum showed that compared with Group S，a dramatic increase in thickness of the mesothelium-to-muscle layer of peritoneum in Group N, P, R and GW(P＜0.05). Compared with group P, the thickness significantly decreased in Group R(P＜0.05). PET results showed that compared with Group S, ultrafiltration (UF) significantly reduced in Group P, R, and GW(P＜0.05). Compared with Group P, ultrafiltration  significantly increased in Group P, R, and GW (P＜0.05). Compared with group S, the expressions of AQP1, VEGF-A and COX-2 mRNA and protein were significantly increased in group P, R and GW(P＜0.05). Compared with group P, the expressions of AQP1, VEGF-A mRNA and protein were significantly decreased in Group R and GW(P＜0.05). Compared with group P, the expressions of COX-2 mRNA and protein were significantly decreased in group R (P＜0.05), while no differences in the expression of COX-2 mRNA and protein in group GW (P＜0.05).     Conclusions    Rosiglitazone can inhibit peritoneal interstitial and vascular proliferation, protect peritoneal function and increase ultrafiltration. Rosiglitazone can protect peritoneal function probably by inhibiting expression of VEGF-A and COX-2.

Objective    To investigate the  role  of  silent mating type information regulation 2  homologue 1(SIRT1) in renal ischemia-reperfusion(IR) injury and its effect on NF-κBp65 - peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) signal pathway in mice.    Methods    Seventy-two healthy C57BL/6 male mice were randomly divided into four groups: control group(n=18), sham-operated group(n=18), IR group(n=18), resveratrol group(n=18). Bilateral renal pedicle were clamped for 45 min was adopted to establish the model of acute ischemic renal injury, to give 2% dimethyl sulfoxide or resveratrol by intraperitoneal injection for 7 days before modeling. Determination techniques included routine biochemical methods for the the levels of Scr and BUN, spectrophotometry for the level of superoxide dismutase (SOD), HE staining for the histological changes as well as immunohistochemical method and Western blotting for the expressions of SIRT1, NF-κBp65 and PGC-1α, respectively.    Results    Compared with that in control and sham-operated groups, the levels of serum Scr and BUN were higher and SOD levels in renal tissues were lower at 12 h and 24 h after operation in IR groups(P＜0.05). HE staining revealed evident pathological lesions including necrosis of renal tubular epithelial cells in IR group. Compared with that in IR group, resveratrol attenuated the above-mentioned changes. Western blotting revealed the up-regulated SIRT1 expression and the activated NF-κB signal pathway, the up-regulated p65 expression and the down-regulated PGC-1α expression subsequent to IR(P＜0.05). Both Western blotting and immunohistochemistry showed that the expressions of SIRT1 and PGC-1α in resveratrol group were up-regulated compared to that in IR group(P＜0.05), while the NF-κBp65 expression in resveratrol group was down-regulated(P＜0.05). Conclusions    In mouse model of renal ischemia-reperfusion injury, the activation of SIRT1 can inhibit the NF-κBp65 expression and accordingly up-regulated PGC-1α level, contributing to inhibiting inflammatory reactions and attenuating oxidative stress-induced injury in the protection of the kidneys.

Objective    To investigate the relationship of α-klotho protein and obesity related glomerulonephritis.    Methods    The chronic kidney disease (CKD) patients with or without ORG were diagnosed by renal biopsy. The normal and abdominal obesity control people were enrolled from the physical examination center. Propensity scoring analysis was done to balance the four groups of people in important clinical characteristics. The α-klotho levels in blood and urine were detected by ELISA. ORG mouse model was established and the mRNA and protein expression of klotho protein were detected by real-time quantitative PCR and Western blotting.    Results    (1) The plasma α-klotho levels decreased in ORG patients, CKD patients and abdominal obesity control people compared with normal control people [(251.7±124.1) ng/L, (336.3±126.1) ng/L, (377.1±120.4) ng/L vs (472.3±204.2) ng/L, all P＜0.05]. The ORG patients had the lowest plasma α-klotho levels (P＜0.05). (2) ORG patients also had the lowest urine α-klotho levels compared with CKD patients, abdominal obesity and normal control people [(24.7±11.4) mg/mol vs (82.5±33.8) mg/mol, (74.5±32.5) mg/mol, (100.8±51.1) mg/mol, all P＜0.05]. There was no difference in urine α-klotho levels of CKD patients, abdominal obesity and normal control people. (3) Compared with the normal control mouse, ORG model mouse showed decreased mRNA and protein expression of α-klotho protein in renal tissue.    Conclusion    The lower plasma and urine α-klotho levels in ORG patients may be due to the reduced expression of α-klotho protein in kidney.

Objective    To investigate the effect of globular adiponectin on the high expression of monocyte chemotactic protein-1 (MCP-1) induced by high glucose in rat renal tubular epithelial cells(NRK52E), and its relationship with adiponectin receptors and p38MAPK.    Methods    NRK52E cells were cultured in vitro and divided into six groups: normal glucose group (NG, 5.6 mmol/L glucose), high glucose group(HG, 25 mmol/L glucose), gAd group1 (HG+gAd 2 mg/L), gAd group2 (HG+gAd 5 mg/L), gAd group3 (HG+gAd 10 mg/L), p38MAPK antagonist group：(SB, HG+SB203580 10 μmol/L). The protein expression of phosphorylated p38MAPK (p-p38MAPK), total p38MAPK (t-p38MAPK),   MCP-1 and AdipoR1/AdipoR2 were examined by western blotting. The mRNA expression of MCP-1 and AdipoR1/AdipoR2 were detected by RT-PCR and real-time PCR respectively.    Results    Compared with NG group, the mRNA and protein expression of MCP-1 increased significantly in HG group (all P＜0.05). The phosphorylation of p38MAPK increased (P＜0.05) with no change in t-p38MAPK protein. The addition of gAd or SB203580 inhibited the unregulation of MCP-1 and p-p38MAPK induced by HG. Two kinds of adipoR，adipoR1 and adipoR2，were all detectable in NG group, and mRNA and protein expression of adipoR1 was higher than that of adipoR2 (P＜0.01). Compared with NG group, the expression of adipoR decreased in HG group, but the difference had no statistical significance(P＞0.05). Compared to HG group, the mRNA and protein expression of adipoR1 increased in gAd groups (all P＜0.01).    Conclusion    The gAd can dose-dependently attenuate the overexpression of MCP-1 induced by high glucose, and this protective effect may be mediated by adipoR1 and p38MAPK.