Objective To establish a relatively concise semi-quantitative pathological scoring and approach to predict the prognosis of IgA nephropathy (IgAN). Methods One hundred and fifty-five cases diagnosed as primary IgA nephropathy with 2 years follow-up were enrolled into our study. In order to examine intra- and inter-observer reproducibility of classification according to the scores, pathological data of 91 cases were reviewed twice by one pathologist, and data of 56 cases were reviewed again by another pathologist. Eight histological indices were analyzed at the beginning: (1) endothelial proliferative index (endoI). (2) active crescents and segmental necrosis of glomerular capillary wall (dGAI). (3) mesangial hypercellularity index (MsHI). (4) increment of mesangial matrix area (MsMI). (5) glomerular chronicity index (GCI). (6) interstitial inflammatory cells infiltration (infI). (7) tubular atrophy and interstitial fibrosis (TCI). (8) vascular chronicity index (VCI). Results Twenty-five (16.13%) patients progressed into irreversible end stage renal disease (ESRD) during follow-up period [(69.07±28.66) months, ranged from 10 to 170 months]. Serum creatinine at biopsy was（112.18±83.13） μmol/L. The initial histological variables were analyzed using Cox proportional hazard model and three variables, including dGAI，GCI and TCI, were finally chosen. GCI and TCI were added up to indicate the chronicity index (CI). dGAI and CI were both found as independent factors in predicting renal outcome (RR=1.255 and 1.691, P<0.05, respectively). Patients in group with both higher dGAI (≥4) and CI (≥6) had the worst renal prognosis (P<0.01). For patients in CKD grade 1 and grade 2, multivariate analysis including clinical and histological variables showed CI was the only independent risk factor of bad prognosis. The reproducibility of the scoring system was proved acceptable (inter- and intra-observer’s kappa values >0.4). Conclusions This relatively concise pathological scoring method containing dGAI and CI has good reproducibility and can predict renal outcome of IgAN.

Objective To analyze the clinicopathological features of IgA nephropathy (IgAN) patients with anemia and the influencing factors of prognosis. Methods The clinical and pathological data of patients diagnosed with primary IgAN at the First Affiliated Hospital of Fujian Medical University from January 1, 2006 to December 31, 2016 were retrospectively analyzed. The patients were divided into anemia group and non-anemia group according to whether the patient was anemia or not. The clinical and pathological data of the two groups were collected. All of them were followed up from the date of renal biopsy to January 1, 2018. Survival curves of the two groups were drawn by Kaplan-Meier method, and compared by Log-rank test. Multivariate Cox proportional hazards regression model was adopted to explore the influencing factors of prognosis in IgAN patients. Results A total of 231 subjects were enrolled, including 122 males (52.8%), and the male-female ratio was 1.12∶1. Their age was (34.8±10.1) years (15-68 years). There were 70 patients (30.3%) in anemia group, 161 cases (69.7%) in non-anemic group. Compared with non-anemia group, anemia group had higher proportion of females, lower serum albumin, higher proportion of tubular atrophy/interstitial fibrosis (T1/2), endothelial cell proliferation (E1) and crescent formation (C1/2), which were statistically significant (all P＜0.05). The patients had a median follow-up time as 6.3 years (0.3-12.9 years). Survival analysis showed that patients in anemia group had lower cumulative renal survival rate than that in non-anemia group ( χ2=15.234, P＜0.001). Multivariate Cox hazards regression analysis revealed that anemia (HR=3.820, 95%CI 1.674-8.719, P=0.001), tubular atrophy/interstitial fibrosis (T1/2) (HR=3.770, 95%CI 1.026-13.852, P=0.046), glomerular segmental sclerosis/adhesion (S1) (HR=4.211, 95%CI 1.139-15.576, P=0.031), hypertension (HR=2.988, 95%CI 1.276-6.999, P=0.012), increased 24 h urinary protein (HR=1.103, 95%CI 1.046-1.163, P＜0.001) and estimated glomerular filtration (eGFR)＜60 ml?min-1?(1.73 m2)-1 (HR=3.725, 95%CI 1.639-8.462, P=0.002) were the independent risk factors for poor renal prognosis in patients with IgAN. Conclusions The clinicopathological features of IgAN patients with anemia are relatively serious, and the renal cumulative survival rate is lower. Anemia, tubular atrophy/interstitial fibrosis (T1/2), glomerular segmental sclerosis/adhesion (S1), hypertension, increased urinary protein and eGFR＜60 ml?min-1?(1.73 m2)-1 are the independent risk factors for poor renal prognosis in patients with IgAN.

Objective To establish an optimal experimental animal model of IgA nephropathy (IgAN). Methods Three kinds of proteins, Dextran G200, outer membrane proteins (OMPs) of Bacillus coli (E.coli) and 20 peptide cellular antigen determinant of staphylococcus aureus, were used to induce IgAN animal model respectively. Animal models were identified and their clinicopathological features were compared by molecular biological and pathological methods. Results (1) In dextran G200 group, urinary protein elevated obviously accompanied by hematuria; immunofluoresence staining revealed a great quantity of IgA depositing in some glomeruli; light microscopy showed a remarkable proliferation of mesangial cells in kidney and diffuse pink-dyed substance deposition in liver and spleen; electron microscopy demonstrated a low electron dense deposition in glomerular mesangial region and amylonloid material in liver and spleen. (2) In OMPs group, urinary protein elevated obviously accompanied by hematuria; immunofluoresence staining revealed less amount of IgA deposition in glomeruli; light microscopy showed a slight proliferation of mesangial cells and obvious interstitial infiltration of inflammatory cells; no electron dense deposition was found in glomerular mesangial region by electron microscope. (3) In 20 peptide cellular antigen determinant of staphylococcus aureus group, urinary protein elevated obviously accompanied by hematuria as well; immunofluoresence staining revealed a large quantity of IgA deposition in most glomerui; light microscopy showed a slight proliferation of mesangial cells and matrix; electron microscopy demonstrated a high electron dense deposition in subendothelial cells and mesangial region. Conclusion The clinicopathological characteristics of IgAN animal model induced by 20 peptide antigen determinant of staphylococcus aureus are similar to human IgAN, which is the best among these three animal models.

Objective To investigate the clinical and pathological characteristics and prognosis of IgA nephropathy (IgAN) patients with uniformly thin glomerular basement membrane (TGBM)， as well as the relationship between IgAN with TGBM and thin glomerular basement membrane disease (TGBMD). Methods According to systematic measurement of GBM thickness by electron microscope (EM)， the IgAN patients were divided into two subgroups: IgAN with normal-GBM， and IgAN with TGBM. Based on the clinicopathological data， the clinical and pathological analysis and the linkage analysis with COL4A3/COL4A4 gene in three IgAN with TGBM pedigrees were performed. Microsatellites PAX3 and HaeⅢ-RFLP， closely linked to the COL4A3 and COL4A4 loci at 2q35-37， were used as genetic linkage markers. Family genetic linkage analysis was performed with the Population and Pedigree Analysis Programs. Results The normal thickness of GBM was (352.43±32.11) nm， and the TGBM(205.56±23.48) nm in IgAN patients. (1) 31.4% (21/66) patients were accompanied with TGBM in the familial IgAN (FIgAN)， while only 11% (24/219) in the sporadic IgAN. (2) The clinical manifestations of 30 IgAN patients with TGBM were generalized as following: predominant in the affected female， persistent haematuria， the high incidence of familial history with renal disease， the less quantity of urinary protein and benign clinical courses. (3) Linkage analysis in three IgAN with TGBM pedigrees indicated that two of them were linked with PAX3， HaeⅢ-RFLP makers of COL4A3/COL4A4 gene respectively， and the LOD score was 1.53 (θ=0.00). Conclusions The clinical features of IgAN patients with TGBM are different from those with normal GBM. Some families of IgAN with TGBM may converge with the families of TGBMD， so the electron microscopy examination is necessary for familiar IgAN.

Objective To elucidate the clinical features of nutcracker syndrome complicated with IgA nephropathy (IgAN) and to increase its level of diagnosis and treatment. Methods Clinical data of 14 cases of nutcracker syndrome complicated with IgA nephropathy (patient group) and 36 cases of nutcracker syndrome (control group) were analyzed retrospectively. Nutcracker syndrome was diagnosed by ultrasonography and magnetic resonance angiography (MRA) and IgAN by renal biopsy. Differences of clinical data and images in two groups were analyzed. Results Gender, age and blood pressure of two groups were not significantly different. Higher Scr level [(81.2±21.3) μmol/L vs (61.2±11.8) μmol/L, P<0.01], more severe proteinuria [(1.1±0.6) g/d vs (0.3±0.2) g/d, P<0.01] and hematuria (2.3±0.9 vs 1.5±1.3, P<0.05) in patient group were found. Differences of ultrasonography and MRA in two groups were not significant. Conclusion Renal biopsy should be considered in cases of nutcracker syndrome with persistence of proteinuria, hematuria or abnormal morphology of urinary red blood cell.

Objective To investigate the clinicopathological characteristics of IgA nephropathy (IgAN) with urinary podocyte excretion（UPE）. Methods (1) Among the 36 enrolled patients with IgAN, 20 were male and 16 were female, with an average age of (34.1±12.2) years. All these patients were diagnosed by renal biopsy. Ten healthy volunteers were served as controls. (2)The UPE level was calculated by urinary sediment smear counting. (3) The correlation between UPE level and renal pathological changes was examined. Results (1)The patients with IgA nephropathy had a higher frequency of podocyturia (61%). No podocyturia was found in healthy controls. (2)The UPE level, as well as UPE/urinary creatinine or UPE/urinary renal epithelial excretion ratio, were significantly elevated in IgAN patients with massive proteinuria (≥3.0 g/24 h). Moreover，the UPE level was positively correlated with 24 h urinary protein excretion（r＝0.446，P＝0.007）. (3)Compared with negative staining patients, patients with positive podocalyxin stain showed significantly higher 24 h urinary protein excretion and urinary renal epithelial cells excretion, while their plasma albumin levels were significantly lower. These two groups had similar distribution in age, gender and frequency of hypertension, and no significant variations were observed concerning the levels of Scr, Hb, cholesterol and triglyceride（P > 0.05）. (4) UPE was related to the injuries of cellular crescent, glomerular disruption of capillaries and extensive epithelial foot process effacement, but not related to the mesangial proliferation, or focal capillary wall thickening. Patients with glomerular crescents showed more severe glomerular sclerosis and interstitial fibrosis (P<0.05). Conclusions Podocyturia is not only a clinical evidence of renal damage, but also a hallmark of the activity of IgAN. Urinary podocyte excretion level in IgAN patients is correlated with 24 h urinary protein excretion and specific pathological damages.

Objective To investigate the injury of podocyte and its association with proteinuria in IgA nephropathy (IgAN). Method Thirty-five patients of IgA nephropathy with proteinuria more than 1.0 g/24 h were enrolled in the study, and eight cases of renal harmatomaectomy or renal cancinomaectomy were as controls. Cell cycle regulatory proteins (p21, p27), podocyte-associated molecules (integrin-β1, nephrin, α-actinin 4, nestin), foot process width (FPW) and the amount of podocyte were examined by immunohistochemistry and real-time PCR, respectively. Patients were divided into two groups according to podocyte number per volume (Nv): podocytopenia group (n=15, Nv<52.49×106/μm3) and normal number group (n=20, Nv≥52.49×106/μm3). Proteinuria was followed up for eighteen months. Results Compared with the controls, podocyte p21 was re-expressed, while the expression of p27 was decreased (0.71±0.12 vs 0.91±0.07, P<0.05) in IgAN. The nestin protein level was markedly decreased in IgAN(13.4%±0.04% vs 17.6%±0.04%, P<0.05). The mRNA expression of integrin-β1 was significantly increased (12.54±5.20 vs 1.02±0.30, P<0.05), while the amount of nephrin, α-actinin4 remained unchanged. Effacement of foot processes and podocyte detachment from the glomerular basement membrane were observed in some cases. Nv was significantly less than that of controls (161.27±225.92 vs 323.22±138.12, P<0.05）， which was associated with the Lee’s grade of IgA nephropathy. The integrin-β1 mRNA expression and Nv were negatively correlated with baseline proteinuria by univariate analysis (r=－0.840, P=0.034; r=－0.4483, P=0.014, respectively). Proteinuria in podocytopenia group was decreased more slowly than that in normal number group. Conclusions Podocyte injury exsists in IgAN with proteinuria, which manifests alterations in cell cycle regulatory protein and some podocyte-associated molecules, as well as foot process effacement and loss of podocyte. Podocyte injury may be involved in proteinuria by affecting the progression of proteinuria in IgAN.

Objective To assess the efficacy of fish oil in the treatment of IgA nephropathy using the method of Meta-analysis. Methods Randomized controlled trials of fish oil in the treatment of IgA nephropathy were searched in the database of Cochrane library, PubMed, EMBASE and CNKI. Data extracted from the literatures were analyzed with Revman software (version 5.0). Results In comparison with the controlled group, proteinuria in the fish oil group was significantly decreased [SMD=-0.27, 95%CI(-0.52 to -0.03), P=0.03], while the renal function deteriorated [SMD=0.30，95%CI(0.05 to 0.55), P=0.02]. Conclusion Fish oil can decrease the proteinuria of IgA nephropathy, but can not prevent renal function from deterioration.

Objective    To explore the clinical significance of complement activation in IgA nephropathy (IgAN) patients and provide new potential therapy targets.    Methods    Biopsy-proven IgAN patients admitted in our renal center were retrospectively recruited. Demographic, baseline clinical and pathological data were recorded as well as the follow-up results. Patients were divided into three groups, negative, weak positive and strong positive group, according to the intensity of C3 deposition in mesangial area of glomurili. Decreased serum C3 level was defined as C3＜85 mg/dl. Results    In this study, 528 IgAN patients were recruited and mean follow-up time was 3 years. There were 119 (22.5%), 164(31.1%), 245(46.4%) patients in the negative, weak positive and strong positive group respectively; 93(21.7%) patients had decreased serum C3 level and 335(78.3%) patients had normal serum C3 level; Significant negative correlation was found between mesangial area of C3 deposition and serum level of C3(r＝-0.209, P＜0.01). The age or sex were similar among different groups of mesangial C3 deposition. In univariate analysis, higher baseline serum creatinine, uric acid and IgA levels, and lower estimated glomerular filtration rate(eGFR), body mass index (BMI) levels were associated with a higher grade of mesangial C3 deposition (P＜0.05). Endocapillary hypercellularity and tubular atrophy or interstitial fibrosis were more prominent in patients with higher grade mesangial deposition of C3. Compared with the patients with normal serum C3 level, patients with decreased serum C3 level had lower white blood cells, hemoglobin, triglyceride, cholesterol, eGFR level and higher serum creatinine level (P＜0.05).  During the follow-up, a total of 54 patients developed to end stage renal disease (ESRD), the incidence of ESRD was 23.7% in patients with decreased serum C3 level and 8.4% with normal C3 level. Kaplan-Meier analysis showed that median outcome-free survival time of patients with decreased serum C3 level was significant shorter than patients with normal serum C3 level [(145.0±22.5) months vs (150.8±17.0) months, P＜0.01]. Cox regression proportional hazards models showed that after adjusting by sex, age and clinical indicators (MAP, eGFR, serum albumin, urine protein and hemoglobin level),  decreased serum C3 level (HR＝0.97, 95%CI 0.96, 0.99, P＜0.01) remained be an independent risk factor of ESRD.    Conclusions    There are different levels of complement activation in patients with IgAN. Complement activation is associated with baseline renal function and clinical outcomes, and decreased serum C3 level is an independent risk factor of ESRD in IgAN patients. Complement activation may be involved in the progression of IgAN.

Objective To investigate somatic mutation of C1GALT1C1, a coding gene for molecular chaperone cosmc of β1,3 galactosyltransferase, in patients with IgA nephropathy(IgAN). Methods Twenty-seven IgAN patients and 19 normal healthy controls were enrolled in the study. Firstly, the coding region of C1GALT1C1 was amplified by PCR from genomic DNA in peripheral blood and PCR products were directly sequenced. Meantime, DNA from peripheral blood B lymphocyte was extracted from 15 IgAN patients and 7 normal controls. The coding region of C1GALT1C1 was amplified from DNA in peripheral blood B lymphocyte, and then, PCR products were subcloned into PGEM-T vector. Total 202 clones, including average 8 to 10 clones per individual, were randomly selected for directly sequencing. Results T393A polymorphism in the coding region of C1GALT1C1 was found in 2 patients and 1 control from their peripheral blood genomic DNA. The minor allele frequency was 6.9%, which was a bit lower than that reported in dbSNP (9.5%). No mutations or polymorphisms were found in total 202 clones from B lymphocyte DNA in 22 persons (15 patients and 7 controls). Conclusions A polymorphism, T393A, in the coding region of C1GALT1C1 gene is identified in present study. Minor allele frequency is lower than that in previous report. Somatic mutation can not be found in B lymphocyte of patients with IgAN.

Objective To study the molecular pathologic features of CD71 and correlation between IgA deposits and CD71 expression in IgA nephropathy(IgAN),and to investigate the role of CD71 in pathogenesis of IgAN. Methods According to clinical manifestations and pathological diagnosis, renal tissue samples of 120 cases were divided into IgA deposit nephritis (the primary group) 44 cases, non-IgAN with IgA deposit (the secondary group) 38 cases and no IgA deposit nephritis (the control group ) 38 cases. Immunofluorescence double-staining with TRITC-conjugated anti CD71 antibody and FITC-conjugated anti-human IgA antibody was performed to illustrate the expression of CD71 and IgA respectively under confocal microscopy. Results The expression of CD71 in the IgAN tissues was consistent with the accumulation of IgA. Significant differences of the expression of CD71 and IgA in glomeruli were found between primary and secondary group. The expression levels of CD71 and IgA were much higher in the　primary group than those in secondary group. Moreover CD71 expression was not found in control group. IgA and CD71 were co-localized under confocal fluorescence microscopy. Conclusion CD71 is highly expressed in glomeruli of primary IgAN and co-localized with IgA, which may play an important role in immunopathogenesis of IgAN.

Objective To examine the correlation of urinary podocyte number and giomerular podocalyxin expression with clinicopathology in IgA nephropathy (IgAN) patients. Methods Morning urinary specimens (100 ml) 3 days before renal biopsy from 50 patients with IgAN diagnosed by renal biopsy and from 20 healthy volunteers as control were collected. After centrifugation, 300 μl sediment was used for smear. Immunohistochemical staining with monoclonal anti-podocalyxin antibody was performed to detect urinary podocytes and the number of podocyte was counted under optical microscope. Computer image analysis system was used to examine glomerular PCX expression. Renal pathology and classification were investigated based on Lee’s grading and Katafuchi semi-quantitative integration method. Relevance analysis was carried out on urinary podocyte number, glomerular PCX expression with pathological score and clinical data. Results The amount of urinary podocytes in IgAN was obviously higher than that in healthy controls (P<0.01). Significant differences were found in multiple comparison of the median of urinary podocytes among Lee’s grade groups. Ⅰ-Ⅱ group was lower as compared to Ⅲ, Ⅳ, Ⅴ groups (all P<0.05). Ⅲ group was lower as compared to Ⅴ group (P<0.05). The positive rate of urinary podocyte was the highest in Ⅳ and Ⅴgroups (100%), while the lowest in Ⅰ-Ⅱ group (55%). Glomerular PCX expression in IgAN decreased with the aggravation of renal pathology. Significant differences were found in multiple comparison of the glomerular PCX expression with the pathological score. Lee’sⅠ-Ⅱ group was higher as compared to Ⅲ, Ⅳ, Ⅴ groups (all P<0.05). Ⅲ and Ⅳ groups were higher as compared to Ⅴ group (P<0.05). In IgAN, urinary podocyte excretion was negatively correlated with glomerular PCX expression (r=-0.702, P<0.01), positively correlated with 24-hour urinary protein (r=0.465, P<0.01) and positively correlated with glomerular and tubular scores (r=0.233, 0.307, P<0.05). Glomerular PCX expression was negatively correlated with 24-hour urinary protein (r=-0.367，P<0.05) and negatively correlated with glomerular and tubular scores (r=-0.560,-0.377, P<0.05). Conclusions Injury and desquamation of glomerular podocytes may involve in the development of IgAN. The number of urinary podocyte can reflect the loss of podocytes in renal tissue, which may be used as a marker of disease progression of IgAN.

Objective To investigate the effects of CD4+CD25high regulatory T cells (Treg) and the imbalance of helper T lymphocyte subsets(Th1/Th2) on the immunological mechanism of IgA nephropathy (IgAN) patients. Methods The percentage of Treg and helper T cells subpopulation (Th1/Th2) in the peripheral blood of IgAN patients and healthy controls was examined by flow cytometry. The FOXP3 expression was detected through intracellular staining. The correlation of Treg or Th1/Th2 with clinical parameters of IgAN was analyzed by Spearman or Pearson rank correlation test. Results The percentages of Treg and Th2 cells were significantly higher in peripheral blood of IgAN patients compared to that of healthy controls[Treg(2.14±0.82）％ vs（1.59±0.53)%, Th2（2.57±0.72）％ vs（1.81±1.10）％, all P＜0.05]. Th1/Th2 ratio was significantly reduced in IgAN patients （5.75±1.89 vs 12.73±9.79， P＜0.05）. The percentage of circulating Treg cells was positively correlated with serum IgA concentration (r=0.397, P＜0.05), and was negatively correlated with eGFR（r=－0.376, P＜0.05）. The percentage of circulating Th2 cells was positively correlated with serum IgA（r=0.468, P＜0.05）. Conclusions There is a disorder of T lymphocyte population in the peripheral blood of IgAN patients. The increased Treg and Th2 cells may play an important role in the pathogenesis of IgAN.

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.

Objective To explore the clinicopathological features of IgA nephrolpathy associated with malignant hypertension (IgAN-MHT) and to analyze their correlation with renal vascular lesions. Methods Twenty-nine patients of IgAN-MHT were screened from 2000 biopsy-proven cases with primary IgA nephropathy (IgAN) in our department from April 1997 to May 2007. Data of clinicopathology and follow-up of these 29 patients were collected. Semi-quantitative analysis was performed to evaluate the pathological changes. Inner lumen, outer lumen, intimal thickness, tunica media-to-internal lumen ratio of 436 arterioles, 124 interlobular arteries and 5 arcuate arteries were measured. The primary endpoint was the composite of a doubling of serum creatinine level and ESRD. Correlations of renal vascular lesions with clinical manifestation, pathological change and prognosis were examined by Spearman and Cox methods. Results 1.5% of all the IgAN patients presented malignant hypertension. The common clinical features were renal failure (100%), hyperuricacidemia (62.7%) and hypertriglyceridemia (51.7%). The average amount of urine protein excretion was 2.8 g/d. The common pathological changes were moderate mesangial proliferation, severe global sclerosis, severe interstitial inflammation and severe interstitial-tubular fibrosis. The small arteries (arcuate arteries and interlobular arteries) and arterioles (afferent arterioles) were both involved in IgAN-MHT. The characteristic lesions of intrarenal arteries included vascular occlusion, media thickening, proliferative endarteritis (onionskin lesion, musculomucoid intimal hyperplasia), hyaline arteriosclerosis, but mainly vascular occlusion (86.2%). The arteriole lesion was negatively correlated with age and total protein level; vascular occlusion was positively correlated with uric acid level. The average follow-up period was 21.1 months. Forteen patients reached the endpoint. The arteriole lesion was the main independent risk factor for the progression of IgAN-MHT (RR=10.21, 95%CI=1.16~89.67). Conclusions The main clinical feature of IgAN-MHT is renal failure. The main histological feature of intrarenal vascular lesions is occludes arterioles. Arteriole lesion is the main independent risk factor for the progression of IgAN-MHT.

Objective To evaluate the clinical value of detecting serum underglycosylated IgA1 in diagnosis and differentiation of IgA nephropathy(IgAN). Methods 　Serum underglycosylated IgA1 was isolated by microspincolumn coupled with vicia villosa lectin(VVL) from 48 cases with IgAN and 43 cases with other primary glomerulonephritis. All the patients were diagnosed by renal biopsy. Sera from 20 healthy persons were used as control group. After isolation, the eluant with rich underglycosylated IgA1 was detected by incubation with biotin-labeled horseradish peroxidase(HRP) and Helix aspersa（HAA, recognizing N-acetylgalactosamine specifically）in enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity of diagnosis and differentiation of IgAN with elevated serum underglycosylated IgA1 were analyzed. Results The level of serum underglycosylated IgA1 in IgAN patients [(83.7±41.0) U] was significantly higher than that in healthy control group [(52.6±22.9) U] and the patients with other primary glomerular diseases[(49.2±27.3) U] (all P<0.01). Twenty-two cases of non-IgA mesangial proliferative glomerulonephritis accounted for 51% of other primary glomerular disease, whose underglycosylated IgA1 level [（47.6±21.5） U] （all P<0.01） was significantly lower as compared to IgAN patients. Taking the renal biopsy diagnosis as golden diagnostic criteria, the ROC curve was performed. The area under the curve was 0.797 with a standard error 0.047(P<0.01). The sensitivity as a diagnostic test was 72.9%, with specificity 72.1% and accuracy 72.5%. Conclusion Detection of serum underglycosylated IgA1 level by microspincolumn method and ELISA assay has certain clinical value in diagnosis and differentiation of IgAN.

Objective To clarify whether the Cosmc gene down-regulation in IgA nephropathy (IgAN) patients is resulted from genetic disorders or external suppressions. Methods Forty IgAN patients, 16 non-IgAN glomerulonephritis patients and 21 healthy controls were enrolled in the study. Genomic DNA was extracted and then Cosmc gene was amplified and sequenced. Peripheral B lymphocytes were isolated and cultured with RPMI-1640 alone or plus lipopolysaccharide (LPS). The Cosmc mRNA expression levels at baseline, after RPMI-1640 culture or RPMI-1640+LPS treatment were measured respectively by real-time RT-PCR. Results (1) Only 2 missense mutations and 2 silent mutations were detected in coding frame region of Cosmc gene in 2 IgAN patients. (2) The baseline Cosmc gene expression level was significantly lower in IgAN patients(31% of that in healthy controls) than that in healthy controls. (3) Relative quantification PCR indicated that Cosmc mRNA expression level was significantly increased (219% of baseline) after RPMI-1640 culture, and treatment of LPS could strongly inhibit this effect. (4) The Cosmc gene expression of healthy control was not affected by RPMI-1640 or LPS. Conclusion It is not genetic disorders but external suppression to cause the down-regulation of Cosmc gene mRNA expression in IgAN.

Objective To analyze prognosis of pregnancy and kidney disease, and evaluate effects of renal pathology on pregnant outcomes and clinical risk factors of adverse outcomes of pregnancy in IgA nephropathy (IgAN) patients. Methods IgAN patients with more than 20 weeks of pregnancy were included, by retrieving the medical database in Peking Union Medical College Hospital from January 1996 to December 2015. Their detailed information during hospitalization and follow-up was recorded, and outcomes of pregnancy and kidney diseases in IgAN patients were assessed. According to Lee's renal pathological grade system, patients were divided into gradeⅣ&Ⅴ group and below grade Ⅳ group to compare their pregnant prognosis. IgAN patients were divied into fetus survival group and fetus death group according to their pregnancy outcomes. The fetal survival factors were analyzed by single factor and multivariate regression. Results A total of 64 pregnancies in 62 patients were included with a mean age of (30.31±4.05) years. The fetus survival rate was 87.5% and the average gestational periods was (35.41±5.10) weeks (ranging from 20-40 weeks). The incidence of pregnancy-induced hypertension syndrome is 17.2% (11 cases). The preterm birth rate was 24.1% (14 cases) among the live births. Serum creatinine increased in 18 cases (28.1%) during pregnancy with median increment of 38.5 μmol/L, and 72.2% patients completely recovered to the level before pregnancy in the postpartum period of 6 months. The incidence of fetus death (38.1% vs 0.0%, P＜0.01), low birth weight infant (46.2% vs 11.1%, P＜0.05) and pregnancy-induced hypertension syndrome (33.3% vs 11.1%, P＜0.05) in Lee's grade Ⅳ&Ⅴ group was higher than those in below grade Ⅳ group. The serum creatinine, urine protein excretion, renal hypertension before pregnancy and renal segmental glomerular sclerosis were significantly increased in fetus death group as compared with those in fetus survival group (all P＜0.05). Logistic regression showed that in all patients an estimated glomerular filtration rate (eGFR)＜60 ml?min-1?(1.73 m2)-1 (OR=76.978, 95%CI 3.327-1780.939, P=0.007) and renal hypertension (OR=14.464，95%CI 1.245-168.053, P=0.033) before pregnancy were the independent risk factors for fetus death, while multipara was a protective factor (OR=0.063, 95%CI 0.005-0.876, P=0.040). Conclusions The fetus survival and kidney prognosis in IgAN patients are closely related to the severity of clinical and pathological changes before pregnancy. Reduced eGFR and complication of renal hypertension are the independent risk factors for adverse prognosis of pregnancy.

Objective To study the prognosis of IgA nephropathy(IgAN). Methods Forty-six out of 780 IgAN patients biopsied in a single unit of north China since 1997 showed segmental necrotizing lesions. Thirty-five of these patients were followed up for (26±26) months after biopsy. Their morphological features and natural history were compared with those of control group of 80 patients without segmental necrosis， who had comparable serum creatinine with a follow-up for (39±23) months. Progression was indicated as 50% elevation of serum creatinine or 33% decline of Ccr or development of ESRD. Their clinical outcomes were compared using Kaplan-Meier estimation. Cox regression was performed to identify predictive factors for clinical events. Results Only 2 necrotizing patients showed ARF. No difference was found in the clinical symptoms presented. The necrotizing group showed a more significant accumulation of monocytes and lymphocytes (P = 0.004) with lower percentage of glomerular sclerosis (P = 0.002). Among the groups of follow-up shorter than 24 months， 3 of 22 necrotizing IgAN patients and 1 of 28 non-necrotizing IgAN patients had futher progression (P = 0.08). In other groups， all necrotizing IgAN patients were event-free， 13 of 52 non-necrotizing IgAN patients progressed， in which necrotizing lesion was not an independent risk factor. Multivariate analysis showed that lesions with severe chronic background were the only prognostic factors (RR = 23.13， P < 0.01)， indicating prognosis was associated with the severity of universal lesions， not with necrotizing lesions alone. Conclusions Based on the present data， necrotizing lesion is not an independent risk factor and may not require aggressive therapy. Longer period of follow-up is needed for further confirmation.

Objective To initially map the gene responsible for autosomal dominant familial IgA nephropathy of a Chinese family by exclusive the five loci that had been reported with linkage analysis. Methods The genetic pattern of the familial IgA nephropathy was identified and the genomic DNA was extracted from the blood samples collected from the family members. Short tandem repeat (STR) inside the loci that had been reported was selected, such as 2q36, 3p23-24, 4q26-31, 6q22-23, 17q12-22, and the data with two-point linkage analysis were performed. Results Autosomal dominant inheritance pattern was demonstrated in phenotypes of the family and there was no linkage relationship in the above five loci of chromosomes because the maximum two-point LOD score was 0.39 at D17S1868. Conclusion Following exclusion of the loci which had been reported, there are other new pathopoiesis loci of FIgAN and it reveals that FIgAN has the genetic heterogeneity according to initial result at the same time.

Objective To elucidate the clinical and pathologic characteristics of IgA nephropathy associated with ankylosing spondylitis（AS）. Methods Clinical and pathologic data of 10 patients suffered from IgA nephropathy associated with AS were reviewed. They were admitted to our hospital from 1997 to 2006 and diagnosed by renal biopsy. Results The average age of nine male and one female patients was (28.6±6.8) years old. Four patients presented with asymptomatic hematuria, and six with hematuria and proteinuria. All the patients suffered from microhematuria and two from macrohematuria. The average 24-hour proteinuria was （1.56 ±1.53） g. Two had hypertension. Serum creatinine levels of all the cases were normal. Light microscopy examination showed that eight patients were mild mesangial proliferation with Lee’s classification grade Ⅰ or grade Ⅱ, and the other two were moderate to severe mesangial proliferation with grade Ⅲ or grade Ⅵ. Conclusion Mild mesangioproliferative IgA glomerulonephritis may be the major morphological pattern in AS patients with renal involvement.

Objective To search the susceptibility candidate megsin gene and P-selectin gene of IgA nephropathy by detecting single nucleotide polymorphism (SNP) and to provide important data for subsequent case-control association study. Methods A comprehensive survey was performed on 12 sporadic IgAN patients and 12 normal controls ，who were from Shanghai， Jiangsu and Zhejiang Han populations， by direct sequencing. 13 591 bp of genomic sequence， including entire coding region， promoter region， part of regulatory region， and exon-intron connection region， was assayed to detect SNPs. Results After the results of sequencing were analyzed， 11 SNPs in SERPINB7 gene and 16 in SELP gene were identified in a total of 27 SNPs， with an average frequency of 1SNP per 503 bp. Ten SNPs， 37% of 27 SNPs， had not been reported in GenBank. Conclusion Our findings not only provide important data for subsequent case-control association study，but also are beneficial to genetic variation bank of Chinese population.

Objective To investigate the influences of Epstein-Barr virus transformation on the expression of IgA1 molecular glycosylation associated gene ST6GALNAC2 and the alteration of its encoded protein 2,6-sialyltransferase. Methods Two patients with IgA nephropathy, 2 patients with IgA multiple myeloma and 3 healthy controls were enrolled in the study. Peripheral blood samples from participants were used for B lymphocytes enrichment by CD19+ immunomagnetic beads and for EBV transformation. The expression of gene ST6GALNAC2 was detected by real-time RT-PCR and the level of sialic acid in IgA1 molecule was observed by ELISA. Results The expression level of gene ST6GALNAC2 in B lymphocytes after EBV transformation was lower than that enriched from fresh peripheral blood. The level of sialic acid in IgA1 molecule secreted from B lymphocyte was down-regulated after EBV transformation. Conclusion In the investigations on the expression of IgA1 molecular glycosylation associated genes and the effect of its encoded enzymes, B lymphocytes enriched from fresh peripheral blood can not be entirely replaced by B lymphocyte transformed by EBV.

Objective To analyze the changes of clinical and pathological features in the patients of IgA nephropathy with anemia. Methods Four hundred and nine patients of IgA nephropathy diagnosed by renal biopsy were classified into two groups: IgA nephropathy with non-anemia (group 1) and IgA nephropathy with anemia (group 2). Changes were studied retrospectively between the groups. Results Serum hemoglobin level was correlated with the clinical parameters of IgA nephropathy. Compared to group 1, changes in group 2 were as followed: serum creatinine increased, eGFR decreased, proteinuria increased; the global sclerosis, segmental sclerosis, crescents and tubulointerstitial lesions worsened. The glomerular and tubulointerstitial lesions were negatively correlated with serum hemoglobin and eGFR, but positively correlated with serum uric acid and proteinuria (P<0.05). Multivariate Logistic regression analysis revealed that anemia was an independent risk factor for the tubulointerstitial lesion. Conclusion Clinical feature and pathological damages in the patients of IgA nephropathy with anemia are more serious than those with non-anemia.

Objective To investigate the relationship of mannose-binding protein(MBP) gene polymorphism to various patterns of glomerular immune deposits in IgA nephropathy patients of Uygur nationality in Xinjiang. Methods Sixty-eight patients with biopsy-proven IgA nephropathy were identified from renal disease database. Two hundred healthy donors served as normal controls. MBP codon 54 gene polymorphism was detected by PCR-RFLP. MBP genotype and allele frequency were compared between different patterns of glomerular immune deposits in IgA nephropathy patients and normal controls. Results Distribution of MBP codon 54 gene polymorphism showed no signficant difference between Uygur IgA nephropathy patients and Uygur normal controls. Distribution of MBP gene polymorphism in Uygur IgAN patients with different patterns of immune deposits showed significant difference. The genotype frequency of GAC heterozygote was significantly higher in group with glomerular IgA, IgG, IgM, C3 and C1q deposits in the mesangium as compared to that in group with isolated IgA and C3 deposits (44.7％ vs 20.0％), and as well as the GAC allele frepuency(0.303 vs 0.133). Conclusions Immunopathological polymorphism of Uygur IgAN is influenced by genetic context. The GAC heterozygote of MBP gene codon 54 may contribute to different patterns of glomerular immune deposits in Uygur IgAN patients

Objective To analyze the clinicopathologic features of proliferative sclerosing IgA nephropathy and the efficacy of prednisone therapy. Methods A retrospective analysis was conducted, enrolling 50 patients with biopsy-proven primary proliferative sclerosing IgA nephropathy who were admitted in the Hospital from January 2005 to June 2015 - 26 males and 24 females, mean age (36.8±10.4) years. Clinicopathologic features and prednisone therapeutic effect were analyzed. Results The clinical manifestations of 50 cases were nephritis syndrome with varying degrees of renal insufficiency, including 32 cases (64.0%) with hypertension, 15 cases (30.0%) with microscopic hematuria. Renal biopsy showed the incidence of glomerular global sclerosis was 17.0%-47.2%, tubular atrophy/ interstitial fibrosis outstanding (T0 50%, T1 32%, T2 18%). After prednisone treatment, compared with sustained remission group and relapse group, invalid patients had higher incidence of hypertension (P＜0.05), relatively lower Hb (P＜0.01) and serum albumin, more significant renal dysfunction (P＜0.01), more severe glomerular global sclerosis, segmental sclerosis, tubular atrophy/interstitial fibrosis, while the lower interstitial inflammatory cell infiltration. During the follow-up, which lasted from 6 to 132 months (median 27.3 months), the effective rate of treatment was 74.0% after sufficient prednisone or half dose prednisone therapy. Repeated recurrence rate was 32.0%. At the end of the follow-up period, 13(26.0%) patients entered the stage of uremia. Conclusions Application of glucocorticoids in the treatment of proliferative sclerosing IgA nephropathy can protect renal function and delay the progression of renal impairment. The efficacy of glucocorticoids therapy is significantly associated with the presence or absence of hypertension, the degree of renal function impairment, and the severity of the onset of renal pathology.

Objective To investigate the blood pressure circadian rhythm in patients with IgA nephropathy by ambulatory blood pressure monitoring and explore its role in the disease progression. Methods A cross sectional study was carried out. Blood pressure rhythm was studied by ambulatory 24-hour monitoring with a portable oscillometric recorder in selected patients with primary IgA nephropathy. The term "dipper" was described as blood pressure during night dropped at least 10% below daytime blood pressure． The term "non-dipper" referred to those in whom the nocturnal decline in blood pressure was less than 10%． Clinicopathological indices between dipper and non-dipper groups were compared. Results Ninety-three patients completed ambulatory blood pressure monitoring among whom 68(73%) patients were non-dipper. The frequency of non-dipper was 70%, 70% and 81% in the patients at chronic kidney disease stage 1，2 and 3 or more. The frequency did not differ among these three group patients (P=0.587). 77% of patients with hypertension and 69% of patients with normotension were non-dipper (P=0.373). The disappearance of blood pressure circadian rhythm in IgA nephropathy was not influenced by age, gender, blood pressure, proteinuria, renal function and renal pathology lesions. Among the patients who were followed up regularly for more than 12 months (n=54), patients in the dipper group had a trend of slower eGFR decline rate than those in non-dipper group albeit the difference was not significant (P=0.329). Subgroup analysis revealed that in patients with hypertension and non-dipper (n=29), the eGFR decline rate was much faster than that in dipper group[(-6.79±11.58）vs (-0.34±1.74) ml&#8226;min-1&#8226;(1.73 m2）-1&#8226;year-1, P=0.019]. Conclusions Most patients with IgA nephropathy present disappearance of blood pressure circadian rhythm, even among those at an early stage or without hypertension. The loss of blood pressure rhythm may be associated with a rapid renal function decline rate in those with hypertension.

Objective    To investigate the clinical and pathological characteristics of IgA nephropathy (IgAN) with macrohematuria (MH).    Method    1512 consecutive patients with biopsy-proven IgAN diagnosed from January 2006 to December 2011 were enrolled, and divided into MH group and control group respectively, according to whether there existed episodes of MH before renal biopsy. The clinical and pathological characteristics were compared between two groups. Patients in MH group were then divided into three groups according to the interval from the last episode of MH to renal biopsy to clarify the concomitant clinicopathological changes associated with occurrence of MH.    Results    The rate of MH in history was 22.1%. MH group patients had significantly lower serum creatinine, slighter proteinuria, lower prevalence of hypertension and heavier microhematuria than control group (all P＜0.001). The prebiopsy durations were similar in two groups (P=0.627). In MH group, chronic pathological indicators, including global/segmental sclerosis, tubule atrophy/interstitial fibrosis were all slighter (all P＜0.001), whereas activity indicators, including necrosis lesions, crescents and mesangial proliferation were all more severe compared with control group (all P＜0.05). Those who underwent renal biopsy within 30 days of the last episode of MH had more severe proteinuria and microhematuria, higher prevalence of necrosis lesions, more severe crescents formation, and endothelial proliferation (all P＜0.05).    Conclusions    IgAN patients with MH in history have relatively milder clinical and chronic pathological manifestations, however more active pathological changes especially in those who suffer episode of MH recently.

Objective To investigate the effects of supernatant of cultured mesangial cells with serum IgA1 from IgA nephropathy patients on apoptosis of podocyte. Methods Jacalin affinity chromatography and Sephacryl S-200 molecular sieve chromatography were used to isolate IgA1. Apoptosis rate of podocyte was assessed by flow cytometer. Monomeric IgA1(mIgA1) was transformed to aggregated IgA1(aIgA1) by heating. IgA-mesangial cell supernatant was prepared by collecting spent medium in which growth-arrested mesangial cells were incubated with different aIgA1, then the medium with RPMI 1640 containing 0.5%FBS was cultured with growth-arrested podocyte. Real time PCR was used to detect the mRNA expression of Bcl-2, Bax, Fas and Fas-L. Results Apoptosis rate of podocyte by supernatant of cultured mesangial cell with aIgA1 from IgAN patients was higher than that from healthy and control groups [(28.5±5.9 ) % vs (22.5±5.8)%, (20.5±4.5)%, all P<0.05]. Fas mRNA expression of podocyte exposed to supernatant of cultured mesangial cells with aIgA1 from IgAN patients increased significantly and was 1.89 folds of control (P<0.05), while Bcl-2 mRNA expression significantly decreased and was 72% of control (P<0.05). The concentrations of AngⅡand TGF-β1 in supernatant of cultured mesangial cells with IgA1 from IgA nephropathy were significantly higher than those from healthy control [(13.2±3.4) ng/L vs (8.2±2.3) ng/L, P<0.05; (15.4±3.4 ) ng/L vs (10.8±3.2) ng/L, P<0.05]. Conclusion Supernatant of cultured mesangial cells with IgA1 from IgA nephroapthy patients can induce apoptosis of podocyte, which may play a role in the progression of IgAN.