Objective To observe whether bone marrow mesenchymal stem cells (MSCs) can promote the repair of IgA nephropathy and to explore its possible mechanism. Methods Sprague-Dawley rats were randomly divided into three groups which were MSCs injection group, normal saline(NS) infusion group and healthy control group. IgA nephropathy model was established by the improving method with BSA +SEB +CCl4 in former two groups. MSCs of SD rats were continuously cultured in vitro and identified with specific surface antigens by flow cytometry and osteogenic and adipogenic differentiation. MSCs were labeled with bromodeoxyuridine (BrdU) in vitro before transplanted. At 1st and 4th week after MSCs injection, the changes of body weight, urine protein, renal function, histopathology and IgA immunofluorescence were observed. MCP-1, TGF-β1 in urine were detected by ELISA. The expression of MCP-1, TGF-β1 in kidney were examined by RT-PCR. The cytokines and BrdU labeled MSCs were detected by immunohistochemistry to observe the disposition in kidney. Results At the end of the first week of MSCs transplantation, MSCs group urine protein (36.86±4.78) mg/24 h, serum creatinine (53.50±6.28) μmol/L, and the NS group urine protein (66.98±5.86) mg/24 h, serum creatinine (82.50±8.36) μmol/L, the differences between two groups were significant (P<0.05). At the same time, the content of MCP-1, TGF-β1 in urine and expression in renal tissue of MSCs group were obviously less than those of NS group (P <0.05). At the end of the 4th week, the body weight, histopatholngy, IgA immunofluorescence of MSCs group were remarkably improved as compared with those of NS group. The content of MCP-1, TGF-β1 in urine and expression in renal tissue, and renal pathological change in MSCs group had no significant differences as compared with those of healthy control group. As the time passed, the disposition of BrdU-labeled MSCs in kidney was taper. Conclusions MSCs injection contributes to renal repair in rat IgA nephropathy. The mechanism may partly depend on adjusting the excretion of cytokines in renal microenvironment and/or other functions rather than completely depend on their differentiation to renal cells.

Objective To explore the role of phospholipase A2 receptor 1 (PLA2R1) in the diagnosis,differential diagnosis and evaluation of idiopathic membranous nephropathy (IMN) in adult patients.Methods A total of 242 renal disease patients diagnosed by renal biopsy from March 2015 to January 2016 were enrolled,consisting of 90 IMN,20 secondary membranous nephropathy (SMN),82 IgA nephropathy (IgAN),30 minimal changed disease (MCD),16 focal segmental glomerulosclerosis (FSGS) and 4 membranoproliferative glomerulonephritis (MPGN).Their clinical data including age,sex,serum creatinine (Scr),serum albumin and 24 h urinary protein were collected.Serum PLA2R1 was measured by enzyme linked immunosorbent assay.PLA2R and IgG subclasses in glomeruli were detected by indirect immunofluorescence assay.The positive rate of serum PLA2R1 among those groups and its correlation with clinical-pathological parameters were analyzed.Results Compared with IMN patients,SMN,MCD and FSGS patients were younger (all P ＜ 0.01); IgAN patients were younger and had higher serum albumin and lower 24 h proteinuria (all P ＜ 0.001); MPGN patients had higher Scr (all P ＜ 0.01).The positive rate of serum PLA2R1 was 75.6％ in IMN patients,while it was 0.0％ in non-IMN patients.The distribution between serum PLA2R1 and pathological diagnosis had difference (P ＜ 0.001),their positive coincidence rate was 100％,negative coincidence rate was 87.4％,total coincidence rate was 90.9％ and their consistency was well (Kappa=0.795,P ＜ 0.001).Among IgG subtype comparisons between IMN patients and SMN patients in the glomeruli,only moderate or more positive IgG4 had statistical differences (82.2％ vs 5.0％,P ＜ 0.001); the positive rate of glomerular PLA2R1 was 41.1％ in IMN patients,higher than 10.0％ in SMN patients (P=0.009); positive PLA2R1 with moderate or more positive IgG4 in glomeruli in IMN patients was more than that in SMN patients (40.0％ vs 0.0％,P ＜ 0.001),which could improve the diagnostic specificity of IMN.In IMN patients serum PLA2R1 and glomerular PLA2R1 had statistical differences (P＜0.001).Spearman rank correlation analysis showed that serum PLA2R1 of IMN patients positively correlated with 24 h proteinuria (r=0.315,P=0.002),negatively correlated with serum albumin (r=-0.228,P=0.030) and didn't correlate with Scr (r=0.199,P=0.059).Conclusions Serum PLA2R can be used as the specific indicator for diagnosis,differential diagnosis of IMN and to reflect the severity of IMN in patients.

Objective To test the hypothesis that advanced glycosylation end products(AGEs) increase cellular inflammation in atherosclerotic plaques. Methods Fifty rabbits were randomly divided into five groups. Hypercholesterolemic (0.5% cholesterol in diet) rabbits received repeated intravenous injection of either AGEs modified rabbit serum albumin (AGEs-RSA ) (group A) or unmodified RSA (group B) for 10 weeks. Rabbits treated with either hypercholesterolemic diet (group C)or with a normal diet(group D) or with a normal diet, and intravenous injection of AGEs-RSA (group E) were served as controls. Aortas were harvested at the 10th week, and lipid deposition was quantitated by oil red 0 staining. Macrophage (RAM-11 positive cells) and T lymphocyte (CD43 positive cells) infiltration, smooth muscle cell(?-actin positive cells) migration and proliferation were determined by using immunohistochemical staining and image-analysis techniques. Results Atherosclerotic plaques could be found in animals fed with hypercholesterolemic diet.Lipid deposition in plaque was significantly higher in group A (71.86%?8.3%) than those in group B (53.76%?3.72%)and group C (56.67%?9.2%). Infiltrations of macrophage[ (23.1?8.5)/0.01 mm2]and T lymphocyte[ (15.1 ? 3.8)/0.01 mm2]as well as migration and proliferation of smooth muscle cell [ (19.2?5.7)/0,01 mm2] in atherosclerotic lesions were significantly increased in animals treated with hypercholesterolemic diet and received injection of AGEs-RSA (group A) when compared with group B [macrophage (14.4? 5.9)70.01 mm2; T lymphocyte (9.1?2.6)/0.01 mm2; smooth muscle cell (12.9?3.8)/0.01 mm2]and group C[macrophage (15.4?4.4)/0.01 mm2; T lymphocyte (10.5?2.2)/0.01 mm2, smooth muscle cell (13.8?3.9)/0.01 mm2]. Neither plaque nor a cellular inflammation was found in animals fed with normal diet (group D)and in those received repeated injections of AGEs-RSA (group E). Conclusion AGEs increase cellular inflammation in atherosclerotic plaques and may accelerate formation of atherosclerosis in AGEs associated diseases.

OBJECTIVETo analyze the effect of special staining and immunohistochemical staining in distinguishing the types of renal amyloidosis to improve the diagnosis accuracy.

METHODSCongo red staining with different methods, and immunohistochemical staining of Kappa, Lambda and Amyloid A with different antigen retrieval methods were used for staining frozen and paraffin-embedded renal tissue sections.

RESULTSWright's Congo red staining produced a better contrast and a higher resolution and showed a greater stability than the other 2 methods after repeated use for staining the renal tissue sections (P<0.05). Immunofluorescent staining produced better results in frozen renal tissue sections than in paraffin-embedded tissues. Immunofluorescent staining produced had better performance than immunohistochemical staining in paraffin-embedded tissues. The retrieval effect with protein kinase K was the best among the antigen retrieval methods in paraffin-embedded tissues.

CONCLUSIONWright's Congo red staining is better than the other 2 methods in diagnosing renal amyloidosis. Immunohistochemical staining of Kappa, Lambda and Amyloid A in frozen renal tissue sections is necessary to discriminate the types of renal amyloidosis. For paraffin-embedded renal tissues, antigen retrieval using protein kinase K is better than the other 2 methods.

Amyloidosis ; classification ; diagnosis ; pathology ; Coloring Agents ; Congo Red ; Fluorescent Antibody Technique ; methods ; Frozen Sections ; Humans ; Immunohistochemistry ; Kidney ; pathology ; Kidney Diseases ; classification ; diagnosis ; pathology ; Paraffin Embedding ; Staining and Labeling ; methods

OBJECTIVETo determine the levels of urinary soluble intercellular adhesion molecule-1 (sICAM-1) and vascular cellular adhesion molecule-1 (VCAM-1) in patients with lupus nephritis (LN), and explore their correlation with renal disease activity.

METHODSUrine samples were collected from 92 renal biopsy-proven LN patients and 20 healthy controls. Renal disease activity was determined according to the ISN/RPS 2003 Revised Classification of Lupus Nephritis. The urine levels of sICAM-1 and VCAM-1 were detected by enzyme-linked immunosorbent assay, and the expressions of intrarenal ICAM-1 and VCAM-1 were evaluated by immunohistochemisty staining.

RESULTSUrinary sICAM-1 and VCAM-1 levels were elevated in LN patients compared with the controls. Significantly higher levels of urinary sICAM-1 and VCAM-1 were found in patients with active LN, who had also significantly increased intrarenal expressions of ICAM-1 and VCAM-1 compared with patients in remission. A strong positive correlation was noted between intrarenal expression and urine levels of ICAM-1 and VCAM-1. The receiver-operating characteristic (ROC) curve of urine sICAM-1 showed tan area under ROC curve of 0.874 for all participants in the test. A cutoff of 1095.00 pg/mg creatinine yielded a good sensitivity (0.945) and specificity (0.789). The ROC curve of urine VCAM-1 showed an area under ROC of 0.882 for all the participants, and a cutoff of 898.11 pg/mg creatinine yielded a good sensitivity (0.982) and specificity (0.667).

CONCLUSIONUrine sICAM-1 and VCAM-1 levels are positively correlated with their intrarenal expressions and reflect the activity of the nephritis, and therefore they may serve as potential biomarkers for early diagnosis of active LN.

Biomarkers ; urine ; Case-Control Studies ; Early Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Humans ; Intercellular Adhesion Molecule-1 ; urine ; Kidney ; metabolism ; Lupus Nephritis ; diagnosis ; urine ; ROC Curve ; Sensitivity and Specificity ; Vascular Cell Adhesion Molecule-1 ; urine