Objective To evaluate the diagnostic value of pathological features of atypical membranous nephropathy (AMN). Methods Ninety-one patients with AMN diagnosed by renal biopsy during 2011 and 2017 were enrolled in this study. On the basis of M-type phospholipase A2 receptor (PLA2R) and thrombospondin type-1 domain-containing 7A protein (THSD7A) by immunohistochemistry, patients were divided into AMN group (25 cases without PLA2R and THSD7A) and idiopathic membranous nephropathy (IMN) group (66 cases with positive PLA2R or THSD7A). The results of immunofluorescence (IF), light microscopy (LM) and electron microscopy (EM) of these two groups were compared, and the parameters with statistical difference were screened out in order to assess their value in the diagnosis of AMN in fourfold table. Results IF results showed that in AMN group the proportions of IgG deposition on capillary wall and mesangial area as well as positive other IgG subclasses and complement C1q but negative IgG4 were significantly higher than those in IMN group (respectively, 56.0% vs 12.1%, 44.0% vs 0, both P＜0.05). Their diagnostic specificities for AMN were 87.9% and 100.0%, respectively. However, the positive rates of IgG accompanied with IgA and/or IgM, predominant IgG4 with other IgG subclasses and complement C1q in two groups were not significantly different (all P＞0.05). LM results showed that the proportions of false double track sign on basement membrane and fuchsinophilic proteins under epithelium, endothelium, basement membrane and mesangial region in AMN group were significantly higher than those in IMN group (respectively, 36.0% vs 0, 44.0% vs 1.5%, both P＜0.05). Their diagnostic specificities for AMN were 100.0% and 98.5%, respectively. However, the scores of mesangial cell proliferation of these two groups showed no significantly difference (P＞0.05). EM results showed that the rate of endothelial electron dense deposits in AMN group was significantly higher than that in IMN group (36.0% vs 1.5%, P＜0.05), and its diagnostic specificity for AMN was 98.5%. Conclusions IgG deposition on both capillary wall and mesangial area, positive other IgG subclasses and C1q with negative IgG4, false-double contour sign, multi-site fuchsinophilic deposits and endothelial electron dense deposits may help for the AMN diagnosis in the absence of PLA2R and THSD7A related data.

Objective To study the role of C3a and C5a in focal segmental glomerulosclerosis (FSGS) patients. Methods (1) A total of 66 patients with FSGS confirmed by renal biopsy were selected, including 18 cases of tip lesion, 11 cases of perihilar, 22 cases of not otherwise specified (NOS), 10 cases of cellular, and 5 cases of collapsing FSGS. The normal renal tissue resected from patients with kidney tumor was taken as a negative control. The expression of C3a and C5a in renal tissues was detected by immunohistochemistry. (2) Serum and urine samples from these 66 FSGS patients were collected, and serum and urine samples from 10 healthy adult selected from the same physical examination center in the same term were used as normal controls. The levels of C3a and C5a in serum and urine were detected by enzyme-linked immunosorbent assay (ELISA). Results (1) Immunohistochemical results showed that C3a and C5a were deposited in glomerulus of FSGS patients, and no deposition in normal renal tissues. The semi-quantitative score showed that kidney C3a score was significantly correlated with serum creatinine (r=0.547, P＜0.001) and 24 h urine protein (r=0.329, P=0.007) in FSGS patients, and kidney C5a score was also significantly correlated with serum creatinine (r=0.415, P＜0.001) and 24 h urine protein (r=0.414, P＜0.001) in FSGS patients. (2) The levels of serum C3a and C5a in FSGS patients were higher than those in healthy adults (both P＜0.05), but there was no significant difference among the five pathological types (P＞0.05). The levels of urinary C3a/urinary creatinine, urinary C5a/urinary creatinine were higher in FSGS patients than those in healthy adults (all P＜0.05). The levels of urine C3a/urinary creatinine and urinary C5a/urinary creatinine in collapsing FSGS were higher than other FSGS types (all P＜0.01), but there was no significant difference among the tip lesion, the perihilar, the not otherwise specified and the cellular (P＞0.05). (3) Urinary C3a/urinary creatinine levels were significantly correlated with serum creatinine (r=0.774, P＜0.001) and 24 h urine protein (r=0.430, P＜0.001) in FSGS patients, and urinary C5a/urinary creatinine levels were also significantly correlated with serum creatinine (r=0.677, P＜0.001) and 24 h urine protein (r=0.333, P=0.007) in FSGS patients. Conclusion Complement C3a and C5a may be involved in the pathogenesis of FSGS and may be related to the severity of FSGS.

Objective To explore the early predictive value of red cell distribution width (RDW) for contrast-induced nephropathy (CIN) in patients after enhanced computed tomography (CT). Methods A total of 218 patients who underwent enhanced CT between June 2015 and June 2017 at Huizhou Central Municipal Hospital were enrolled in this study. Patients were divided into CIN group and no-CIN group. The diagnostic criteria for CIN is an increase in serum creatinine (Scr) of more than 44.2 μmol/L or 25% of the baseline value within 3 days of contrast agent use. The general information and clinical characteristics in two groups were compared. The risk factors of CIN were analyzed by logistic regression analysis. The receiver operator characteristic curve (ROC) was used to assess the value of RDW for predicting the occurrence of CIN. Results Among 218 patients, 10(4.59%) patients had CIN. In the CIN group age, baseline Scr and baseline RDW were significantly higher, while hemoglobin, baseline estimated glomerular filtration rate (eGFR), red blood cell, white blood cell, albumin, and high-density lipoprotein cholesterol were significantly lower than those in the no-CIN group (all P＜0.05). Binary logistic regression analysis revealed that baseline RDW (OR=2.250, 95% CI 1.031-4.911, P=0.042) and eGFR (OR=0.963, 95% CI 0.928-0.999, P=0.044) were correlated with the occurrence of CIN. ROC analysis confirmed the area under the curve of RDW as a predictor of CIN was 0.798 (P＜0.001). The cut-off value of RDW was 14.5%, and the diagnostic sensitivity and specificity in CIN were 70.00% and 85.58%, respectively. Conclusions Increased baseline RDW and decreased eGFR are the risk factors of the occurrence of CIN after enhanced CT. RDW has a good predictive value, and it may be a good biomarker for the early diagnosis of CIN.

Objective To evaluate the clinical outcomes of hemodialysis patients after superficial femoral artery-superficial femoral vein arteriovenous graft (AVG). Methods Hemodialysis patients with mid-thigh superficial femoral artery-superficial femoral vein AVG from August 2015 to March 2018 in department of vascular surgery, Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine were enrolled. Their clinical outcomes and complications after operation were analyzed retrospectively. Patency rates were measured by Kaplan-Meier survival curve. Results A total of 18 cases were enrolled. The success rate of the operation was 100% without complication. Follow-up time was (22.00±11.77) months with 100% follow-up rate. The 6 months-, 12 months-, 24 months-primary patency rates were 83.3%±8.8%, 48.5%±12.1%, 24.2%±13.5%, respectively; secondly patency rates were 100.0%, 100.0%, 87.5%±11.7%. There were 1 case of seroma, 1 case of puncture site infection, 11 cases of stenosis and 5 cases thrombosis during follow-up, while no heart failure, ischemia or pseudoaneurysm. Conclusion Mid-thigh AVG has low infection rate and high patency rate, so it can be as the first choice for the lower extremity AVG in hemodialysis patients.

Objective To observe the expression of parathyroid hormone-related peptide (PTHrp) receptor in tibial growth plate and its effects on tibial extension in chronic renal insufficiency rats. Methods Two-week-old male Sprague-Dawley rats were randomly divided into sham group, model group and enalapril group, each with 20 rats. In model group and enalapril group rats had chronic renal insufficiency induced by left ureteral obstruction, and rats were respectively given saline and enalapril by gavage after the operation. In sham group, left ureter was only exposed without ligation, and rats were given saline. The urine was collected 4 weeks after the operation and the total protein content was measured. Then all rats were killed. The concentrations of PTHrp, creatinine and urea nitrogen in intracardiac blood were detected. HE staining and Masson staining were performed on the left kidney to observe pathological changes of glomeruli and renal tubules. The total length of bilateral tibia was measured. The number of columnar cells in the growth plate proliferative zone was measured by safranin O staining and the expression of PTHrp receptor in the growth plate was detected by immunohistochemistry. Results The 24 h urine total protein, creatinine and urea nitrogen in model group were higher than those in sham group (all P＜0.05), while these 3 renal functional parameters in enalapril group were lower than those in model group (all P＜0.05). In model group and enalapril group rats had higher blood concentrations of PTHrp than that in sham group (all P＜0.05), but blood PTHrp in enalapril group was lower than that in model group (P＜0.05). HE staining and Masson staining showed that in the model group rats had severe tubular dilation, inflammatory cell infiltration and the tissue fibrosis, while in enalapril group renal tubules slightly dilated and had a few inflammatory cell infiltration and tissue fibrosis. Compared with those in the sham group, in model group the tibia length, the chondrocyte number of column structure in the growth plate proliferative zone and the PTHrp receptor decreased (all P＜0.05). But in enalapril group those indexes increased than model group (all P＜0.05). Conclusions Chronic renal insufficiency rats had increased PTHrp concentration in the blood but decreased PTHrp receptors expression in tibial growth plate, which lead to their limited tibial extension.

Objective To investigate whether elevated parathyroid hormone (PTH) levels could induce endothelial-to-mesenchymal transition (EndMT) and adipocyte transition in endothelial cells (ECs), and to determine the possible underlying mechanism. Methods (1) A rat model of secondary hyperparathyroidism and chronic kidney disease (CKD) was established. The adiposity in bone marrow was detected by oil red O staining. Immunofluorescence staining was performed to detect the expression and localization of cluster of differentiation 31 (CD31) and fibroblast-specific protein 1 (FSP1). (2) The human umbilical vein ECs were cultured in vitro. Western blotting was performed to detect protein expressions of EndMT-related markers CD31, FSP1 and α-smooth muscle actin (α-SMA) in interference groups with different PTH concentrations (0, 10-11, 10-9, 10-7 mol/L PTH for 48 h) and times (0, 12, 24, 48 h, 10-7 mol/L PTH), as well as the expression of β-catenin in interference groups with different PTH concentrations. The localizations of CD31, FSP1 and β-catenin were observed by cell immunofluorescence. Protein expressions of adipocytes markers peroxisome proliferator-activated receptor-γ (PPAR-γ) and CCAAT/enhancer binding protein-α (C/EBP-α) by Western blotting and the degree of adipogenesis by oil red O staining were detected after transformed ECs were cultured in adipogenic culture medium for one week. Small interfering RNA (siRNA) was performed to silence β-catenin expression. ECs were divided into control siRNA group, β-catenin siRNA group, PTH+control siRNA group and PTH+β-catenin siRNA group. Protein expressions of CD31, FSP1 and PPAR-γ by Western blotting and the degree of adipogenesis by oil red O staining were determined. Results (1) In vivo, compared with the control, CKD rats had increased adipocytes in bone marrow (P＜0.05), and the co-expression of CD31 and FSP1 in bone marrow ECs. (2) In vitro, PTH significantly inhibited the expression of endothelial marker CD31 and increased the expressions of mesenchymal markers FSP1 and α-SMA in concentration-and time-dependent manners. These indexes in 10-7 mol/L PTH group and 0 mol/L PTH group, in 48 h group and 0 h group showed statistical differences (all P＜0.05). In PTH group ECs with 10-7 mol/L PTH for 48 h showed FSP1 accumulation in the cytoplasm and reduced expressions of CD31, and ECs had higher expressions of PPAR-γ and C/EBP-α as well as the degree of adipogenesis than those in control group (all P＜0.05). Furthermore, PTH enhanced the nuclea β-catenin protein levels in ECs in concentration-dependent. The expressions of β-catenin in 10-7 mol/L PTH group and 0 mol/L PTH group showed statistical differences (P＜0.05). β-catenin expressed in the cytoplasm in control group, while it enter into the nucleus in PTH group. Compared with those in PTH+control siRNA group, the expressions of CD31 and PPAR-γ as well as the degree of adipogenesis decreased in PTH+β-catenin siRNA group (all P＜0.05), while the expression of FSP1 increased (P＜0.05). Conclusions PTH induces ECs- to-adipocytes transition by the canonical Wnt/β-catenin signaling pathway, which might account for bone loss in CKD. Silenced β-catenin expression can inhibit PTH-induced EndMT and adipogenesis.

Objective To observe the differential expression of high mobility group box-1 protein (HMGB1) in renal tissues of heat stroke mice models, and to explore its role in the pathogenesis of heat stroke associated acute kidney injury(HS-associated AKI). Methods According to random number table, 20 healthy male C57BL/6J mice were randomly divided into 2 groups, including normal control (n=10) and heat stroke group (n=10). The mice in heat stroke group were given with a 2-hour-exposure in biological simulation chamber (temperature 41℃, humidity 70%). Heat stroke was defined as anal temperature lasting more than 40 degrees Celsius. A 18F-deoxyglucose nuclide labeled vivo imaging was conducted with micro- positron emission tomography(PET)/computer tomography (CT). Serum creatinine was examined with blood example. In order to evaluate the pathological changes, HE stain was conducted with kidney tissue, and mitochondrial morphological changes in kidney tissue were observed by transmission electron microscopy. The expressions of HMGB1 and apoptosis inducing factor mitochondria associated 2 (Aifm2) were examined by immunohistochemical method, and the levels of HMGB1 and RAGE were examined by Western blotting. The cell apoptosis of renal tissue was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). The metabolomics of kidney tissue in mice were detected by liquid chromatography-mass spectrometry (LC-MS), and the pathway enrichment analysis was carried out by KEEG database. Results (1) The body temperature of the mice in heat shock group was significantly higher than that in normal control group 45 min after model establishment (P＜0.05). The level of serum creatinine in heat shock group was significantly higher than that in normal control group (P＜0.05), and the levels of 18F-deoxyglucose increased in skeletal muscle and visceral tissue of the mice in heat-shock group. (2) HE staining showed hemorrhage in collecting duct and tubular endothelial cell swelling, and mitochondrial swelling and deformation were observed by transmission electron microscopy in kidney tissue of the heat shock group. (3)Immunohistochemical method showed that the levels of Aifm2 and HMGB1 in heat shock group were higher (P＜0.05). (4) Western blotting showed that the levels of HMGB1 and RAGE in heat shock group were higher than those in normal control group (P＜0.05). (5) TUNEL showed that the number of cells with positive stain in kidney tissue of the heat shock group was higher than that in normal control group (P＜0.05). (6) Between normal control group and heat shock group, 136 differential metabolites were detected in kidney tissues. After analysis by KEGG database, pathway abnormalities such as unsaturated fatty acid metabolism disorder may be associated with HS-associated AKI, and many differential metabolites such as adrenic acid may be important regulatory points in the pathogenesis. Conclusion Acute kidney injury is a common complication of heat shock. It may be related to the dysfunction of renal mitochondria and activation of apoptotic pathway caused by systemic hypercatabolism, which may be related to the disorder of unsaturated fatty acid metabolism and activation of HMGB1. Some differential metabolites may be of high value in HS- associated AKI studies.