Typing of amyloidosis in renal biopsies of 205 cases / 中华肾脏病杂志

Objective To identify the prevalence of different amyloid types in renal biopsies.Methods The renal biopsies of 205 patients diagnosed as renal amyloidosis from January 1990 to December 2011 were reassessed.Immunohistochemistry was performed with a penal of antibodies directed against λ-light chain,κ-light chain,amyloid A,fibrinogen,transthyretin,apolipoprotein A1,and lysozyme.Immune electron microscopy and gene analysis were performed when the results of immunohistochemistry were indeterminate.Results Among 205 patients,190 cases (92.7％) were classified as immunoglobulin light chain amyloidosis (AL),1 (0.5％) case as amyloid A amyloidosis and 1 (0.5％) case as fibrinogen A α-chain amyloidosis.The amyloid types of remaining 13 (6.3％) cases were undetermined.In the AL patients,the distribution of λ and κ was 6.6∶1.Conclusion AL is the most common form of renal amyloidosis in China,with a predominant light chain type of λ.

Clinicopathological and genetic feathers of 14 Fabry nephropathy patients / 中华肾脏病杂志

Objective To elucidate the clinicopathological and hereditary characteristics in Fabry nephropathy.Methods The clinical and pathological features of 14 patients with Fabry nephropathy were collected.The activities of α-Gal A were measured in 4 probands.Screenings of GLA mutations were done in 12 patients.Results The ratio of Fabry nephropathy in the patients with renal biopsy was 0.074 ％ (14/19 005),the average diagnostic age was (30.57±9.32) years,male to female ratio was 2.5∶ 1.All the patients had proteinuria,and the median of urine total protein (UTP)was 1.71 g/24 h (0.32-4.71 g/24 h).Two of them got nephrotic proteinuria,5 of them got microscopic hematuria,4 of them got renal function insufficiency.Angiokeratomas was the most common manifestation (10/14),followed by cardiac involvement (6/14).Hypohidrosis and diseases of central neural system could also be seen in these patients.There were 9 classic phenotype,the remaining 5 were variants/renal variants.The family information was collected in 10 pedigrees,6 of them had family histories of kidney disease.Renal pathology showed vacuolization of glomerular visceral epithelial cells was the prominent feature,global and segmental glomerulosclerosis were seen in some patients by light microscopy.The myelin bodies or zebra bodies were identified in podocytes by electron microscopy,but only were found in some podocytes of 2 females.The activity of α-Gal A was decreased compared with the normal range in 4 probands.The mutations of GLA were demonstrated in 11 patients.Three novel mutations of GLA gene were identified,which were all deletion/insertion mutations with classic phenotypes.Most variants carried the mutations located in the buried/partial buried areas of α-Gal A (3/11).The classical phenotype carried GLA mutations as W162X,F169S,S201F,N272K,L310R,while variant phenotype carried GLA mutations as I91T,R112H,Q312H.Conclusions The ratio of Fabry nephropathy in patients with renal biopsy is 0.074％.Angiokeratomas and cardiac involvement are often accompanied with Fabry nephropathy.The genotypes of GLA may have close relationships with the phenotypes of Fabry nephopathy.

A female patient of Fabry disease complicated with thin basement membrane nephropathy and investigation of the kindred / 中华肾脏病杂志

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 Nephro]ogy 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.

Proteins structure change of COL4A4 gene point mutation and its association with phenotype in thin basement membrane nephropathy / 中华肾脏病杂志

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.

Clinical significance and histological origin of glomerular epithelial proliferative lesion in patients with focal segmental glomerulosclerosis / 中华肾脏病杂志

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.

Clinicopathological analysis of Castleman disease with kidney injury / 中华肾脏病杂志

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.

Clinicopathological manifestations of nephronophthisis-medullary cystic kidney disease / 中华肾脏病杂志

Objective To report the clinicopathological features of 2 cases of nephronophthisis-medullary cystic kidney disease (NPH-MCKD). Methods The clinical data and pathological changes of renal biopsy in two patients of NPH-MCKD from our hospital were analyzed, and associated literatures were reviewed simultanously. The clinicopathological featuresand diagnosis of NPH-MCKD were discussed. Results Two adolescent patients were admitted to our hospital for indolent renal insufficiency, polyuria accompanied by polydipsia as first signs.Urine analysis showed low specific density urine, mild proteinuria, and few formed elements in urinary sediments. The ability of urine concentration and acidification was decreased. Familial history of renal disease and extra-renal lesions were not found. Renal ultrasound presented an increased echogenicity with diminished cortico-meduUary differentiation, and multiple small cysts in renal corticomedullary border were identified in one case by computed tomography. Pathological examination of renal biopsy revealed diffuse tubular interstitial lesion which was characterized by the triad of tubular basement disintegration, tubular atrophy with cyst development, and interstitial fibrosis. Some of glomerular sclerosis occurred. Cyst development at the corticomedullary border of the kidneys was the specific feature of NPH-MCKD. Conclusions Young patients with impaired tubular function should be suspected of NPH-MCKD. Renal ultrasound or computed tomography can provide an important clue. Multiple renal cysts at the corticomedullary border identified by renal biopsy can be a diagnostic indication for NPH-MCKD.

Mutation analysis of COL4A3/COL4A4 genes in a family with thin basement membrane nephropathy and focal segmental glomerulosclerosis / 中华肾脏病杂志

Objective To elucidate whether focal segmental glomerulosclerosis (FSGS) is a secondary development of the COL4-linked thin basement membrane nephropathy (TBMN) or the primary FSGS produces thin glomerular basement membrane (GBM). Methods The family members presented microscopic hematuria,increasing proteinuria with the years and a dual pathological diagnosis of FSGS and TBMN was made in the proband.DNA linkage analysis at locus 2q36-37 that contains the COL4A3/COL4A4 genes was performed with polymorphic micmsateilite markers D2S434,D2S279,D2S1370,D2S256 and D2S427.Haplotypes were constructed at the COL4A3/COL4A4 loci for affected and unaffected family members.All exons of COL4A3 and COL4A4 genes were screened for mutations in the proband.Mutation screening was also performed for NPHS1,NPHS2,CD2AP,WTI,TRPC6 and ACTN4 to exclude familial FSGS.Mutation or polymorphism found in the family were examined in 50 healthy controls. Results In this family hematuria segregated with the 55224 haplotype at the COL4A3/COL4A4 locus.G to A substitution at nucleotide 1214 resulting in an glycine being replaced by glutamate (G405E) was demonstrated for the first time in cxon 20 of COL4A4 gene.G4OSE was present in all four members of the family with hematuria but not in the seven unaffected family members nor in 50 healthy controls.A novel polymorphism segregating with hematuria (IVS1-4C＞T in exon 2 ofCOL4A3) was also found which was only present in all four affected family members but not in the seven unaffected family members. No mutations were demonstrated in FSGS associated genes,however,a novel SNP (R268Q),which distributed with the disease ineompletely,was described in the NPHS1 gene coding nephrin,the podocyte slit diaphragm protein. Conclusions In this family,FSGS occurres on the basis of TBMN.Maybe the particular COL4A3/COL4A4 mutation and polymorphism work together to develop proteinuria and eventually leading to FSGS.But whether the mutation and the polymorphism segregating with the disease predispose to develop FSGS in TBMN patients is required further study.

The clinic significance of urinary podocytes in patients with focal segmental glomerulosclerosis / 北京大学学报(医学版)

Objective: To address the significance of urinary podocytes in the diagnosis of human focal segmental glomerulosclerosis(FSGS). Methods: Twelve patients with FSGS and 20 patients with minimal change disease (MCD) were diagnosed by routine renal biopsy, and 8 healthy persons as controls. Morning urinary sediments was collected and centrifuged onto glass slides. Urinary podocytes were identified by immunofluorescent staining of podocyte specific protein Podocalyxin(PCX). The state of podocytes in glomeruli was observed using immunofluorescence. Results: Urinary podocytes were found in 8 out of 12 FSGS patients(66.67%), whereas none of 20 patients with MCD and control had podocytes in their urine. FSGS patients with positives urinary podocytes had prominent manifestation of nephropathy syndrome, whereas no nephrotic syndrome in patients with negative urinary podocytes. Focal absence of the expression of PCX, a marker protein of podocytes in glomeruli was found in FSGS patients, and the locations of absence were consistent with the lesions of focal sclerosis in glomeruli. In contrast, PCX was expressed integrally in MCD patients. Conclusion: Appearances of podocytes in urine of patients with nephropathy may be used as one of the reliable, convenient and unharmful accessorial methods for distinguished diagnosis of FSGS and MCD.

Anti-endothelial cell antibodies(AECA) are associated with disease activity of propylthiouracil induced antineutrophil cytoplasmic antibody (ANCA) positive vasculitis / 北京大学学报(医学版)

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.

Antineutrophil autoantibodies and their target antigens in lupus nephritis / 中华风湿病学杂志

Objective To investigate the target antigens and their clinical association with antineutrophil autoantibodies in patients with lupus nephritis (LN).Methods Sera were collected from 92 renal biopsy proven LN patients.Normal neutrophils and white cells from patients with chronic granulocytic leukemia (CGL) were used as antigens in Western blot analysis to detect autoantibodies in the LN sera.Results ①Using neutrophils as antigens,two bands could be blotted:64 000 (33/92,35 9%)and 50 000 (13/92,14 1%).The prevalence of anti 64 000 autoantibody in patients with positive rheumatic factor was significantly higher than that in patients without (54 5% vs 18 8%, P

Effects of mycophenolic acid on proliferation and collagen I of unstimulated or stimulated vascular smooth muscle cells / 中国实用内科杂志

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

Study on the binding capacity of IgA_1 from patients with IgA nephropathy to human mesangial cells / 中国病理生理杂志

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

The relationship between antiendothelial cell antibodies and anticardiolipin antibodies / 中华肾脏病杂志

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

Anti-glomerular basement membrane autoantibodies in Chinese patients with rapidly progressive glomerulonephritis and their clinical relevance / 中华肾脏病杂志

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.

Detection of Epstein-Barr virus in interstitial nephritis by in situ hybridization / 中华肾脏病杂志

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.

Clinical and pathological charactcristics of ANCA associated systemic vasculitis with immune complex deposition in kidney / 中华肾脏病杂志

Objective To analyze the clinical and pathological characteristics of ANCA associated systemic vasculitis(AASV) with immune complex deposition in kidney and compare with that of pauci-immune AASV. Methods Patients with AASV, admitted in our hospital in last 5 years, were retrospectively studied. The clinical and pathological characteristics were compared between patients with immune complex deposition and patients with pauci-immune deposition. Results There were eitht patients with immune complex deposition (five with IgM deposition, two with IgA deposition and one with IgG deposition) and 32 patients with pauci-immune deposition. There was no significant difference in age, gender, type of ANCA, interval between onset of vasculitis and renal biopsy, clinical manifestations and short-term renal survival rate between the two groups. Patients with immune complex deposition had a higher predromal infection rate ( P

IgA_1 from patients with IgA nephropathy induced release of calcium, up-regulation of TGF-? mRNA expression and secretion of fibronectin in human glomerular mesangial cells / 中国病理生理杂志

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