Objective To develop stable cultures of human umbilical cord vascular endothelial cells(ECs)and smooth muscle cells(SMCs)in order to investigate the effect of mycophenolic acid(MPA)on proliferation and collagen Ⅰ production of SMCs.Methods From Sep.2002 to Sep.2003,ECs were cultured from human umbilical cord veins,and SMCs from arteries.Productions of endothelial cells conditioned medium(ECCM)was obtained from serum free Dulbecco's modified eagle medium(DMEM)with or without MPA(0,0.31,1.25,2.50,5.00,10.00 ?g?mL -1).Proliferation of SMCs was performed with 3H-thymidine incorporation scintillation,and collagen Ⅰ production of SMCs was measured with enzyme-linked immunosorbent assay.Results (1)The 3H-thymidine incorporation increased significantly(P

To investigate the location of Epstein-Barr virus (EBV) in renal tissues of patients with interstitial nephritis(IN). Methods By in situ hybridization. EBER1 was detected in renal tissues of 12 IN patients and 10 patients with minimal change nephropathy (MCN) as control group. Results EBER1 was found positive in 3 renal tissues of IN patients. It mainly distributed in the nuclei of renal tubular cells, infiltration cells and glomerular cells and 10 MCN patients were all negative. Conclusion EBV infection may play an important role in the pathogenesis of IN. In different types of IN, EBV infection may play different role.

Objective To determine the prevalence of autoantibodies against glomerular basement membrane(GBM) in sera of Chinese patients with rapidly progressive glomerulonephritis(RPGN)and to evaluate their clinical relevance. Methods Serum anti-GBM antibodies were detected in 29 RPGN patients by enzyme-linked immunoassay(ELISA)using collagenase solublized human GBM as solid phase ligand. Positive sera were also oonfirmed by Western blot analysis. Results Of the 29 RPGN patients, 5(17%)were positive for anti-GBM autoantibodies. One positive for both anti-GBM autoantibody and ANCA. On Western blot analysis. 23 000～27 000 and 40 000～54 000 polypeptides could be blotted. On direct immunofluoresence there were granular deposits of immunocomplex in capillary loops in three of four. Conclusions The prevalence of anti-GBM antibody mediated RPGN is not rare in China. Using ELISA to detect circulating anti-GBM autoantibodies had been proved to be a more specific and sensitive methods. It is important to detect circulating anti-GBM autoantibodies early for patients with RPGN in order to save time for appropriate therapy.

Objective To investigate the relationship between antiendothelial cell antibodies(AECA)and anticardiolipin antibodies(ACA)in patients with lupus nephritis. Methods 58 sera from patients with lupus nephritis were studied. ELISA technique were used to detect both AECA and ACA, and immunoblotting was performed to determine specific endothelial targets. Results The prevalence of IgG-AECA and IgG-ACA positive were 36.2% and 39.7% respectively in the patients with lupus nephritis. 17 out of 23 patients with ACA had higher titers of AECA, while only 4 out of 35 patients without ACA were AECA positive(P

Objective: To investigate the prevalence of anti-endothelial cell antibodies(AECA) and its possible role in the pathogenesis of propylthiouracil (PTU) induced ANCA positive vasculitis. Methods: Sera from 11 patients with PTU induced ANCA positive vasculitis and 10 patients with PTU induced ANCA but without clinical vasculitis were studied. Soluble proteins from in vitro cultured human umbilical vein endothelial cells were used as antigens and immunoblotting technique was performed to identify the specific target antigens. Results: In patients with PTU induced ANCA positive vasculitis group, 10 of the 11 patients in active phase were AECA positive and 7 of the 10 patients turned to negative in remission. AECA consisted of a group of heterogeneous antibodies. In patients with ANCA positive but without vasculitis, none was AECA positive. Conclusion: AECAs recognizing a variety of antigens could be found in sera from patients with PTU induced ANCA positive vasculitis and they had a much closer association with vasculitic disease activity compared with ANCA.

Objective To detect the proteins structure encoded by COL4A4 gene with different missense mutations of thin basement membrane nephropathy (TBMN) and to analyze the effect of gene mutation on the secondary structure of α4 (Ⅳ) chain and its association with phenotype. Methods A COL4A4-linked TBMN patient with FSGS by a missense mutation (g. 1214G>A resulting in p. G405E) diagnosed by clinical manifestations, family history and renal biopsy examination, as well as two controls (one healthy, one pure TBMN carrying a g. 1550G>A mutation resulting in p. G448S) were enrolled in this study. The fragments of cDNA with the two mutations and that of corresponding cDNA from the healthy control were expressed in E. coll. The secondary structures of recombinant polypeptides were analyzed by circular dichroism (CD) spectroscopy. Results CD spectra of healthy control exhibited a negative peak near 208 nm whereas that of TBMN patient with FSGS exhibited a negative peak near 220 nm. Furthermore, the magnitude of the negative peak of this patient decreased as compared with that of healthy control. CD spectra of pure TBMN control was slightly changed with the negative peak remaining near 208 run and the magnitude slightly decreased as compared with that of healthy control. In addition, the secondary structure of pelypeptide from healthy control was composed of about 1/4 α-helix and 1/4 β-sheet, whereas that from the patient presented about 1/3 α-helix without any β-sheet. The secondary structure of polypeptide from pure TBMN control was almost the same as the healthy control, except a shght reduction of α-helix and a slight increase of β-sheet. Conclusions Although the glycine substitutions exists in the nearby domain of α4 (Ⅳ)chain, the TBMN patient complicating FSGS with severe phenotype and g. 1214G>A mutation and the pure TBMN control with the mild phenotype and g. 1550G>A mutation are revealed with different secondary structures of α4 (Ⅳ)chain. Moreover, the secondary structure change of α4 (Ⅳ) chain is consistent with their corresponding phenotype severity.

Objective To investigate the elinicopathoiogical 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 (493:14) years. They presented edema and proteinuria, with mean urinary protein at (2.79±3.56) g/24 h, including one nephrotie 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.

ObjectiveTo investigate the clinical significance and histological origin of glomerular epithelial proliferative lesion in patients with focal segmental glomerulosclerosis (FSGS). MethodsSeventy-four patients with idiopathic FSGS hospitalized in Peking University First Hospital from Jan. 2000 to Dec.2005 were enrolled in this study. Patients were classified into two groups according to with or without glomerular epithelial proliferative lesion. Estimation of active and chronic pathological scores was carried out using a semi-quantitative grade system by two pathologists. Clinical and pathological characteristics were compared between two groups. Immunohistochemical studies were performed to analyze the histological origin of glomerular epithelial proliferative lesion. ResultsThirty-one patients with glomerular epithelial proliferative lesion showed shorter interval from presentation to biopsy (P<0.05), higher percentage of nephrotic syndrome (NS) (P<0.05), higher frequency of segmental glomerulosclerosis(P<0.05), higher pathological active scores (P<0.05) and lower pathological chronic scores (P<0.05)as compared to 43 patients without glomerular epithelial proliferative lesion. Twenty-nine patients were followed up and renal survival rate in patients with glomerular epithelial proliferative lesion (39.7%) was significantly lower than that in patients without glomerular epithelial proliferative lesion (83.3%) (P=0.049). The frequency of glomerular epithelial proliferative lesion and the serum creatinine (Scr) level at biopsy were independent predictors of ESRD (OR value was 1.204, 1.008 respectively ). Glomerular epithelial proliferative lesion did not express mature podocyte markers including WT-1 and pedocalyxin, but stained positive for PCNA, PAX-2 and CK-8. ConclusionsGlomerular epithelial proliferative lesion represents the pathological change of acute stage and active lesion of FSGS, and also may be the pathological marker of severe clinical presentation and worse renal survival. Glomerular epithelial proliferative lesion may be derived from proliferation of parietal epithelial proliferation or de-differentiated podocytes.

AIM: To examine and compare the ability of serum IgA 1, from both the patients with IgA nephropathy (IgAN) and the healthy control, to bind to human mesangial cells (HMC). METHODS: Serum IgA was isolated with jacalin column, heated to aggregated form (aIgA 1) and labeled with [ 125 I]. Binding capacity of aIgA 1 to primary HMC was evaluated by radioligant binding assay, specificity of binding was determined by competitive inhibition, and relative affinities was compared by cross competitive inhibition. RESULTS: Both aIgA 1 from normal control and patients with IgAN bound to MC in a dose-dependent, saturatable manner, but the binding of aIgA 1 from patients was saturated at approximately 200 pmol while that from healthy was at 400 pmol. The Scatchard analysis revealed a Kd of (8 9?2 1)?10 -8 mol/L for patient's aIgA 1 versus (4 3?1 2)?10 -7 mol/L for normal aIgA 1 ( P

AIM: To study and compare the pathophysiological effects of serum IgA 1 from both the patients with IgA nephropathy (IgAN) and healthy controls on human mesangial cells (HMC). METHODS: Serum IgA 1 was isolated with Jacalin affinity chromatography and heated to form aggregates (aIgA 1). Primary HMC were cultured and passage 3 and passage 4 of the cells were used. Intracellular calcium release was assayed with confocal analysis. Expression of TGF-? mRNA and the content of supernatant fibronectin were tested by RT-PCR and indirect competitive ELISA, respectively. RESULTS: aIgA 1 from patients with IgAN was shown to induce release of intracellular calcium, up-regulation of expression of TGF-? mRNA and secretion of fibronectin in HMC in a similar time-dependent manner as aIgA 1 from healthy controls, but the effects of the former were much stronger and the durations was much longer (P