Objective To investigate the clinicopathological features of Castleman disease with kidney injury. Methods Clinicopathological data of 10 Castleman disease patients with kidney injury from Peking University First Hospital and China-Japan Friendship Hospital were analyzed retrospectively. All the cases received biopsies of lymph node and kidney. Their renal tissues were examined by light microscopy, immunofluorescence and electron microscopy. Results Ten patients were all male with mean age (49±14) years. They presented edema and proteinuria, with mean urinary protein at (2.79±3.56) g/24 h, including one nephrotic syndrome (NS). Hematuria occurred in 8 cases, acute renal insufficiency in 6 cases, hypertension in 4 cases. Most of the patients had fever, fatigue, anorexia, weight loss, increased ESR and CRP, hypergammaglobulinaemia and decreased complements. Other abnormalities included anemia, thrombocytopenia, pleural effusion, hepatomegaly, splenomegaly, hypothyroidism, etc. Two cases demonstrated POEMS syndrome, one presented Sjogren syndrome. The enlargement of multiple cervical, axillary and inguinal lymph nodes were identified in all the patients. The pathological patterns of lymph node were plasma cell type in 4 cases, hyaline-vascular type in 3 cases, and mixed type in 3 cases. Pathological examination of renal biopsy showed thrombotic microangiopathy in 5 cases, crescentic glomerulonephritis in 2 cases, renal amyloidosis, minimal change disease and chronic tubular interstitial nephropathy in 1 case respectively. After immunosupressive reagents or COP therapy, lymph nodes became smaller, systemic symptoms were alleviated, proteinuira was decreased or disappeared, and renal function was recovered in most of patients. Conclusions Castleman disease with kidney injury manifests various symptoms with high prevalence of renal insufficiency and multiple systemic damage. Renal lesions present many patterns of pathological change with a higher frequency of thrombotic microangiopathy. It is necessary to examine the lymph nodes by ultrasound, radiology or biopsy for the patients of renal diseases with multiple systemic symptoms.

Objective To evaluate the detection of alpha 5(Ⅳ) collagen chain of skin basement membrane in diagnosis of Alport syndrome among suspected patients. Methods Data of suspected patients with the detection of alpha 5(Ⅳ) collagen chain of skin basement membrane were retrospectively collected and analyzed from January 2007 to March 2008. Results A total of 254 suspected patients ranged from 1 to 71 years old with an average age of (25.85±17.03) years old were enrolled (male/female ratio, 0.76). There was no significant difference in average age between male and female. Abnormal alpha 5(Ⅳ) collagen chain expression of skin basement membrane was found by indirect immunofluorescense in 19 patients among whom 12 cases were negative and 7 cases were discontinous deposit. These 19 patients were diagnosed as Alport syndrome and the diagnostic rate was 7.5%. Conclusions The diagnostic rate of Alport syndrome by detection of alpha 5(Ⅳ) collagen chain in skin basement membrane is significant and helpful for early and differential diagnosis of Alport syndrome.

Objective To determine the urinary polypeptide patterns of glomerulonephritis by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology. Methods Urinary samples of 29 healthy volunteers and 34 patients with glomerulonephritis, including 10 cases of IgA nephropathy (IgAN), 10 cases of membranous nephropathy (MN), 9 cases of minimal change disease (MCD) and 5 cases of lupus nephritis(LN), were collected and separated by magnetic bead，and were screened for polypeptide patterns with a novel high throughput method, MALDI-TOF MS. Results Under the relative molecular weights 10 000 Da, 85 protein peaks were detected in healthy controls group and 109 protein peaks were detected in glomerulonephritis group. Six peaks of 3371.5 Da, 4026.35 Da, 4085.32 Da, 4116.96 Da, 4126.32 Da and 9527.31 Da were up-regulated，while 8 peaks of 861.28 Da, 1205.41 Da, 1642.52 Da, 1913.15 Da, 1976.52 Da, 2087.74 Da, 2193.47 Da and 3015.57 Da were down-regulated by more than 2 folds (P<0.01) in glomerulonephritis group as compared to healthy controls． Urinary polypeptide patterns in different diseases differed significantly from each other, indicating specific disease pattern of polypeptide excretion. Conclusions MALDI-TOF MS is a fast, convenient and high throughput analyzing method capable of screening some relative specific，potential biomarkers from the urine of glomerulonephritis patients thus it possesses better clinical value．

Objective To evaluate the efficacy and safety of REXEEDTM series highflux dialyzer. Methods A randomized cross-over study of 3 × 3 Latin square was designed based on the surface area of dialyzer membrane (1.5 m2 and 2.1 m2). Seventy-two stable maintenance hemodialysis (MHD) patients from Shanghai Renji Hospital and Ruijin Hospital were enrolled in this study for 3 consecutive weeks. REXEEDTM-15AC, 15UC, 21AC, 21UC dialyzers were used as trial group and APS-15U，BIO-HX100 dialyzers were used as control group. The clearances of urea, creatinine, phosphorus and β2-microglobulin were calculated. Adverse event and adverse reactions were recorded. Results There were significantly higher urea and creatinine clearance in 15AC and 15UC dialyzers as compared to APS-15U dialyzer [(222.07±18.74) ml/min, (220.23±26.26) ml/min vs (199.56±14.21) ml/min; (176.73±16.41) ml/min, (175.22±25.94) ml/min vs (165.42±14.68) ml/min, all P<0.05]. There were significantly higher urea, creatinine and β2-microglobulin clearance in 21AC and 21UC dialyzer as compared to BIO-HX100 dialyzer [(230.59±15.24) ml/min, (233.96±7.06) ml/min vs (203.43±36.66) ml/min; (183.50±25.90) ml/min, (181.05±23.94) ml/min vs (166.25±29.82) ml/min; (111.77±53.42) ml/min, (125.54±51.99) ml/min vs (42.39±4.81) ml/min; all P<0.05]. There was no significant difference of phosphorus clearance between REXEEDTM series dialyzers and control dialyzers. The efficiency of urea clearance and urea reduction ratio could achieve clinical targets in REXEEDTM series. Conclusion REXEEDTM series highflux dialyzers are effective and safe for clinical application.

Objective To explore the method to carry out epidemiological investigation of chronic kidney disease (CKD) in rural area surrounding the Tarim Basin, and to elucidate the prevalence and risk factors associated with CKD among the Uygur adults in Moyu county. Methods A total of 1650 residents (age >18 years) from 15 villages in 3 rural town of Moyu county were randomly selected by using a stratified, multistage sampling. All the residents were interviewed and received physical examination and tested for random spot urine of albumin to creatinine ratio (ACR). Estimated glomerular filtration rate (eGFR) was calculated by modified MDRD equation. eGFR<60 ml&#8226;min-1&#8226;(1.73 m2)-1 was diagnosed as reduced eGFR. The associated factors of CKD were examined. Results Valid data of 1552 subjects were enrolled in the study. After the adjustment of age and gender component, the prevalence of albuminuria and reduced eGFR was 4.5% (95% CI 4.4-4.6) and 1.4% (95% CI 1.4-1.5) respectively. Approximately 5.4% subjects had at least one indicator of kidney damage. Age and hypertension were independently associated with CKD. Conclusions Experience and method of epidemiology investigation of CKD in the rural areas of Uygur are obtained through this study. The prevalence of CKD is 5.4% and the awareness is 12.5% in the Uygur adults of Moyu county. Independent risk factors associated with CKD are hypertension and age.

Objective To investigate the mutations ACTN4 and SYNPO genes promoter in sporadic primary focal segmental glomerulosclerosis (FSGS) and to analyze the role of mutations in FSGS. Methods The study consisted of 82 Chinese primary FSGS, including 39 females and 43 males, ranged from 12 to 76 years old. Seventy volunteers were selected as healthy control group. Genomic DNA was extracted from peripheral blood cells of FSGS patients and hair of patients’ parents by polymerase chain reaction (PCR) and direct sequencing to analyze ACTN4 and SYNPO gene promoter mutations. Mutations were matched with GenBank and TRANSFAC software database (www.ncbi.nlm.nih.gov; www.genometix.de; www.gene-regulation. com). Dual luciferase assay system was used to analyze the promoter region mutations, based on PGL3-Basic vector, pRL-SV40 and PC12 cell line. Hair DNA of novel mutation patients’ parents was sequenced. Expression of alpha-actinin-4 and synaptopodin in patients’ kidney tissue was examined by immunofluorescence. Results Three patients with 1-34C>T, 1-590delA and (1-1044delT)+(1-797T>C)+(1-769A>G) heterozygous mutations were found in ACTN4 gene promoter respectively, and two patients with 1-24G>A and 1-851C>T heterozygous mutations in SYNPO gene promoter respectively. The same mutations were not found in the control group of 70 healthy people. Except one patient accepting her parents’ 1-1044delT and 1-797T>C mutated chromosome respectively, no same mutations were found in patients’ parents. Protein expression of alpha-actinin-4 and synaptopodin was reduced in mutated patients’ kidneys. Except 1-1044delT group, luciferase activity in mutated groups decreased. (1-1044delT)+(1-797T>C)+(1-769A>G) mutation was associated with poor outcome and patient with these mutations progressed to end-stage renal failure. Conclusion Mutations of ACTN4 and SYNPO gene promoters affect gene transcription and protein translation, which may contribute to the onset of sporadic primary FSGS.

Objective To investigate the correlation between serum bone metabolism biomarkers and bone mineral density (BMD) in chronic kidney disease (CKD) patients with different stages. Methods Seventy-eight CKD patients were enrolled in this study and were assigned to different groups according to their creatinine clearance(Ccr). Patients with Ccr≥15 ml/min were divided into 4 groups based on clinical CKD 1-4 stage standard, and those with Ccr<15 ml/min were divided into two groups of hemodialysis (HD) and non-HD. Their levels of serum calcium, phosphorus, alkalinity phosphatase (ALP), urea, Scr, osteocalcin (gla-protein, OC), calcitonin (CT), intact parathyroid hormone (iPTH), osteoprotegerin (OPG) and BMD were detected respectively. Results (1) The serum levels of OPG, iPTH and phosphorus increased significantly in stage 3, 4, 5, respectively (P<0.01), and serum OPG level was elevated to (5.1±1.34) ng/L after HD, which was significantly higher than （3.35±0.76） ng/L before HD (P<0.05). The levels of serum OC, CT, calcium, ALP were not significantly different among patients with different CKD stages, while the level of OC was elevated in HD patients (P<0.05). The femoral WARDS triangle BMD of CKD stage 4 patients decreased to 0.77±0.09, which was less than the value of CKD stage 1 patients (P<0.01), with little influence from hemodialysis treatment. (2) The level of serum OPG was positively correlated with the levels of serum phosphorus, iPTH, OC (r = 0.51, 0.39, 0.36，all P<0.01), and it was negatively correlated with the level of Ccr (r = -0.70, P<0.01). The femoral WARDS triangle BMD was negatively correlated with the levels of iPTH, OC, phosphorus and OPG (r = －0.59, －0.51, －0.45, -0.48, all P<0.05). Conclusions Most of serum bone metabolism biomarkers change according to the declined level of Ccr. Compared with serum phosphorus, the levels of iPTH, BGP and femoral WARDS triangle BMD, serum OPG may be early diagnostic markers of renal osteodystrophy in CKD patients.

Objective To investigate the role of rat organic anion transporter 1（OAT1, SLC22A6) in the renal cellular uptake of AAⅠ and its impact on cellular toxicity. Methods HEK-293 cells were transfected with rat OAT1 cDNA or empty vectors. The over-expression of rOAT1 was confirmed by Western blot analysis and its activity was validated by using para-aminohippurate (PAH) as a probe. Cellular apoptosis was examined by flow cytometery using propodium iodode (PI) and annexin V-FITC staining. Results Concentration- and time-dependent intracellular accumulation of AAⅠ was observed in rOAT1-transfected HEK-293 cells. After treatment with AAⅠ at the concentrations of 40 mg/L, 80 mg/L, 120 mg/L and 160 mg/L, respectively, for 45 min, the intracellular concentrations of AAⅠ in rOAT1-transfected HEK-293 cells were higher than those in controls (P<0.05). After treatment with AAⅠ (120 mg/L for 30 min, 60 min, 90 min and 120 min, respectively, the intracellular concentrations of AAⅠ in rOAT1-transfected HEK-293 cells were higher than those in controls (P<0.05). PAH significantly reduced the intracellular accumulations of AAⅠ in rOAT1-transfected HEK-293 cells. After treatment with AAⅠ at the concentrations of 40 mg/L, 80 mg/L, 120 mg/L and 160 mg/L respectively for 35 min, the intracellular accumulations of AAⅠ in rOAT1-transfected HEK-293 cells that treated with PAH were lower than those that were not treated by PAH. Cellular apoptosis and caspase-3 expression in rOAT1-transfected HEK-293 cells were significantly up-regulated as compared to controls (P<0.05). Conclusion rOAT1 is involved in the cellular uptake of AAⅠ which leads to increased epithelial apoptosis. Further studies are suggested to investigate the role of human OAT in the disposition of AA and its toxicological consequences.

Objective To investigate the effect of high glucose on the expression of liver X receptors (LXRs) and ATP-binding cassette transporter A1 (ABCA1) in human macrophages (THP-1 cell line). Methods THP-1 monocytes were differentiated into macrophages by induction of phorbol 12, 13-dibutyrate (PMA). Surface markers of macrophages were identified by CD68 immunohistochemistry. The macrophages were cultured with different concentration (5.6, 11.1, 22.2 and 33.3 mmol/L) of glucose and different time (0, 0.5, 2, 6, 12, 24, 48, 72 h). Real time PCR and Western blotting methods were used to examine the mRNA and protein expression of LXRs and ABCA1. Results As compared to 5.6 mmol/L glucose, macrophage LXRβ and ABCA1 were decreased significantly at both mRNA and protein levels in dose- and time-dependent manner (P＜0.05). Conclusion Hyperglycemia may play a role in the pathogenesis of arteriosclerosis through the inhibition of LXRs and ABCA1 expression in diabetic patients.

Objective To study the effect of oxymatrine on p-STAT3 and PIAS3 signaling molecule and it’s mRNA expression in the proliferation of the human mesangial cells (HMCs) induced by lipopolysaccharide(LPS) and to explore their relationship. Methods HMCs were divided into three groups: control group, LPS group and oxymatrine group. HMC proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. Type Ⅳ collagen in the supernatants of the cultured HMCs was detected by ELISA at 12, 24, 48 hours respectively. At the same time, the protein and mRNA expressions of p-STAT3 and PIAS3 were measured by Western blot and real-time quantitative RT-PCR. Results The cell proliferation, the mRNA and protein expression of type Ⅳ collagen, p-STAT3 in LPS group were increased compared with the control group (P<0.01), but they were decreased in oxymatrine group(P < 0.01). The expressions of protein and mRNA of PIAS3 in LPS group were decreased significantly compared with control group (P<0.01), but they were increased in oxymatrine group (P<0.01). Conclusion Oxymatrine can down-regulate the expression of p-STAT3 and up-regulate the expression of PIAS3, which plays an important role in the process of LPS-induced HMCs proliferation.

Objective To investigate the effect of pentoxifyccine (PTX) on the pathway of high glucosd-induced expression of CTGF in mesangial cells. Methods Cultured rat mesangial celld were used to study the influence of different concentration of high glucose on the expression of TGF-β, CTGF, p-Smad2/3, Smad7 and FN in different exposure time. Furthermore the effect of high glucose plus TGF-β neutral antibody and different concentration of PTX on the obove expression was evaluated as well. Results High glucose could increase TGF-β, CTGF mRNA and protein expression in mesangial cells (P<0.05) in time- and dose-dependent manner, and at the same time p-Smad2/3 expression increased and Smad7 expression decreased (P<0.05). The blockage of TGF-β could decrease high glucose-induced CTGF mRNA and protein expression by 86.4% and 91.8%. PTX could suppress high glucose-induced CTGF expression in mesangial cells. When the PTX dosage increased, the suppressive effect became more remarkable, but PTX had no influence on the TGF-β expression. Conclusions High glucose up-regulates CTGF mRNA and protein expression mainly through TGF-β-Smads pathway. PTX can suppress CTGF expression effectively, but has no direct inhibition of TGF-β expression.