Objective To explore the relationship between clinical manifestations and pathological changes in diabetic nephropathy (DN) and to assess the predictive power of the pathologic classification for DN established by the Renal Pathology Society in 2010. Methods Patients with type 1 or type 2 diabetes and biopsy-proven DN in the Third Affiliated Hospital of Sun Yat-sen University between January 2004 to June 2014 were enrolled in the present study and were followed-up until 31 December 2014. The outcome was defined as renal end-points including renal replacement therapy and doubling of serum creatinine as well as all-cause mortality. The laboratory and histologic data were analyzed and outcomes were assessed using survival analysis. Results Fifty-seven people enrolled in this study were categorized into Class IIa (n=9), Class IIb (n=9), Class III (n=25) and Class IV (n=14) while no participants belonged to Class I. The changes of Class IIa were slight and those of Class IV were severe both in the clinical data (diabetic duration, blood pressure, estimated glomerular filtration rate, urine protein excretion rate, albumin and hemoglobin) and the pathological data (percentage of global glomerulosclerosis, percentage and scoring of interstitial fibrosis and tubular atrophy, scoring of interstitial inflammation and incidence of large vessel lesions). There were no significant differences between Class IIb and III in the above variables except for the scoring of arteriosclerosis. The mean follow-up duration was 25.9 months. Twenty-five patients (43.9%) reached the renal outcomes and six people (10.5%) reached all-cause mortality. The survival analysis showed that there were significant differences among the renal survival curves of different glomerular classes and of different interstitial and vascular scorings, but not in the survival curves related to all-cause mortality. Conclusions The glomerular classes are not completely associated with renal prognosis. The clinical manifestations and renal outcomes are benign in Class IIa, moderate but similar in Class IIb and III and severe in Class IV. The glomerular classification and interstitial and vascular scorings are associated to renal prognosis while their associations with mortality remain to be verified.

Objective To investigate the effect of urate-lowering therapy on renal function in chronic kidney disease (CKD) stages 2-5 patients with hyperuricemia (HUA). Methods A total of 132 patients of CKD stages 2-5 with HUA between July 2016 and December 2017 in Department of Nephrology of the Second Affiliated Hospital of Anhui Medical University were prospectively and self-controlled analyzed. Serum uric acid (SUA), estimated glomerular filtration rate (eGFR) and other clinical parameters were measured at baseline and after 1-6 months treatment. The patients were divided into group A (CKD stages 2-3a) and group B (CKD stages 3b-5) on the baseline value of eGFR. The changes of SUA and eGFR before and after treatment were compared. According to the level of SUA after 6 months treatment, patients were divided into attainment group (SUA＜360 μmol/L) and nonattainment group (SUA≥360 μmol/L). The difference of renal function in pre-treatment and post-treatment was compared. Multiple stepwise linear regression was used to analyze the relationship among the change of eGFR after receiving 6 months' treatment (deGFR) and SUA level, baseline eGFR and other indexes. Results After 1, 3, 6 months treatment, the average levels of SUA, Scr and urea nitrogen of all patients were decreased significantly while eGFR value was increased significantly (all P＜0.050) than those in pre-treatment period. After six-month-therapy, proteinuria and hematuria were improved significantly in all patients (P＜0.001, P=0.001). Compared with pre-treatment period, both the SUA levels of group A and group B were declined significantly while eGFR had a significant rise after treatment (P＜0.001). The change of eGFR post-treatment in group A was significantly higher than that of group B [(13.64±15.35) vs (8.97±9.79) ml?min-1?(1.73 m2)-1, P=0.044]. At 6 months after treatment, the eGFR value increased markedly in both attainment group and nonattainment group compared with pre-treatment period (P＜0.001). After six-month-therapy, the eGFR value in attainment group was increased more obviously than that of nonattainment group [(13.96±14.64) vs (8.03±9.69) ml?min-1?(1.73 m2)-1, P=0.021]. Multiple stepwise linear regression analysis showed that the baseline eGFR value was an influencing factor of deGFR (b=0.161, P=0.020). Conclusions The renal function of CKD stages 2-5 patients with HUA can be significantly improved by urate-lowering therapy, which can effectively reduce proteinuria and hematuria.

Objective To observe the levels of four bisphenols (bisphenol A, B, S and F) and their correlation with renal function in chronic kidney disease (CKD) patients. Methods Patients with CKD were identified according to Kidney Disease: Improving Global Outcomes (KDIGO) criteria. Sixty-three CKD patients and eleven healthy controls were enrolled. CKD patients were further classified as mild renal injury group (CKD stage 1 and 2, n=30), moderate renal injury group (CKD stage 3, n=19) and severe renal injury group (CKD stage 4 and 5, n=14). The levels of four bisphenols in serum were determined by high performance liquid chromatography (HPLC). The correlation between concentrations of four bisphenols and estimated glomerular filtration rate (eGFR) was assessed by Spearman's rank correlation analysis. The associations of four bisphenols with coronary heart disease, diabetes and hypertension in CKD patients were estimated by binary multivariate logistic regression. Results (1) Four bisphenols were not detected in serum of healthy control. In the mild renal injury group the bisphenol A and bisphenol S were not detected, and patients had 5.24 (5.24, 9.38) μg/L bisphenol B and 0.74 (0.74, 0.74) μg/L bisphenol F. In the moderate renal injury group bisphenol S was not detected, and patients had 2.79 (1.01, 4.53) μg/L bisphenol A, 5.24 (5.24, 5.24) μg/L bisphenol B and 0.74 (0.74, 0.74) μg/L bisphenol F. In severe renal injury group patients had 14.30 (7.97, 18.17) μg/L bisphenol A, 0 μg/L bisphenol B, 23.73 (23.73, 136.59) μg/L bisphenol S and 0.74 (0.74, 1.42) μg/L bisphenol F. The levels of bisphenol A and bisphenol S in severe renal injury group were higher than those in the healthy control group, mild renal injury group and moderate renal injury group (all P＜0.05). Bisphenol B and bisphenol F were not statistically different among four groups. (2) Bisphenol A and bisphenol S were negatively correlated with eGFR (r=-0.779, P＜0.001; r=-0.546, P＜0.001). (3) Among CKD patients, bisphenol A was correlated with diabetes (OR=4.951, 95%CI 1.603-15.294, P=0.005), and bisphenol S was correlated with hypertension (OR=4.466, 95%CI 1.575-12.666, P=0.005). Conclusions CKD patients have a variety of bisphenol compounds, especially bisphenol A and bisphenol S. Bisphenol A and bisphenol S have high levels, and their exposures are correlated with renal function.

Objective To explore the relationship of phospholipase A2 receptor (PLA2R) expression in renal tissue with clinical characteristics, prognosis of idiopathic membranous nephropathy(IMN) patients. Methods 134 patients diagnosed as nephropathy proven by biopsy was selected as subjects of this research, including 98 patients with IMN patients, 10 patients with secondary membranous nephropathy and 26 patients with other renal glomerular diseases. The expression of PLA2R antigen in renal tissue was detected by immuno-fluorescence chemistry staining. Results The positive rate of renal PLA2R expression in IMN patients was higher than that in SMN patients (91.84% vs 40.00%, P＜0.01), whereas there is no expression in other glomerular diseases. The PLA2R negative group were mainly stage I membranous nephropathy, and positive group was mainly in stage II. The distribution of pathological stage between the two groups was statistically significant (P＜0.01). Compared with the positive group, the negative group was manifested with higher eGFR[(115.91±23.32) ml?min-1?(1.73 m2)-1 vs (94.06±27.38) ml?min-1?(1.73 m2)-1, P=0.031], associated with the higher 12-month complete remission rate (87.50% vs 44.07%, P=0.021). Conclusions The expression of PLA2R antigen in renal tissue plays an important role in the diagnosis, disease evaluation and prognosis of IMN. The negative PLA2R in kidney tissue of IMN may indicate a good clinical prognosis.

Objective To view and compare the clinical characteristics of renal tubular acidosis in adults and children. Methods Clinical data of patients with renal tubular acidosis diagnosed by Shandong Provincial Hospital affiliated to Shandong University from Jan 1991 to Sep 2017 were reviewed. The difference and consistency in clinical characteristics of renal tubular acidosis between adults and children were analyzed. Results Data from 206 adults and 60 children were analyzed. 89.81% cases in adults were secondary to other diseases, mainly primary Sjogren's syndrome. Most children patients (81.67%) were idiopathic, others largely originated from inherited metabolic diseases. The most common subtype of both was distal renal tubular acidosis. Proximal renal tubular acidosis was easier to be found in idiopathic renal tubular diseases of children. Chief complaints or starting symptoms were mainly composed of polydipsia with polyuria (41.4%) and fatigue (35.3%). Children were typical of growth retardation, rickets and digestive symptoms. The rate of missed diagnosis and misdiagnosis was 41.4 percent. Routine therapy consisted of healing metabolic acidosis and electrolyte disorders, treating underlying diseases and preventing complications. The majority of patients (95.5%) improved after treatments. Conclusions Renal tubular acidosis possesses various underlying diseases, diverse clinical manifestation and high rate of misdiagnosis. Given the high incident of secondary types, investigation of underlying disease, especially autoimmune diseases such as Sjogren's syndrome, is of great importance in adults. Most children patients suffer from primary renal tubular acidosis. Attention should be paid to them in order to reduce the rate of misdiagnosis and teratogenicity.

Objective To observe the expression of NOD2 and epithelial-mesenchymal transition (EMT) related proteins in podocytes in high glucose environment, and explore the molecular mechanism of NOD2 involved in EMT. Methods The human glomerular podocytes were the subjects of study. α-SMA and Nephrin expressions were detected by immunofluorescence; the mRNA and protein expressions of NOD2, Snail and EMT related proteins (α-SMA , Desmin, E-cadherin, Nephrin) were detected by real-time fluorescence quantitative PCR and Western blotting. The podocytes were stimulated by high-glucose after shRNA interfering the of NOD2 expression, and the expressions of Snail and subsequent EMT-related proteins were detected by Western blotting. Prior to the activation of NOD2 by muramyl dipeptide (MDP), shRNA was used to interfere with the expression of Snail. E-cadherin, Nephrin, Desmin, and α-SMA were detected by Western blotting. Results After 24 hours of high glucose stimulation, PCR and Western blotting results showed that the expressions of NOD2 and Snail were significantly increased; the expressions of epithelial phenotype proteins E-cadherin and Nephrin were down-regulated; and the expressions of interstitial phenotype proteins Desmin and α-SMA were increased (all P＜0.05); while there was no significant change in the hypertonic control group. After interference with NOD2, the abnormal expression of Snail and EMT related proteins were all recovered. After interference with Snail expression, Compared with the MDP group, the protein expressions of E-cadherin and Nephrin were significantly increased (all P＜0.05); the expressions of Desmin and α-SMA were significantly decreased. Conclusions High glucose can induce NOD2 expression in podocytes, and promote podocyte epithelial-mesenchymal transition by upregulating Snail expression. Gene intervention targeting the NOD2/Snail/EMT pathway can reduce high-glucose-induced podocyte injury and may provide new ideas for the treatment of diabetic nephropathy.

Objective To investigate the role and mechanism of macrophage activation induced by exosomes from high glucose-treated renal tubular epithelial cells. Methods (1) The supernatant of renal tubular epithelial cells which were cultured in normal glucose control group (5.5 mmol/L D-glucose) or high glucose group (30.0 mmol/L D-glucose) for 48 h were collected and ultracentrifuged to harvest exosomes. Exosomes were identified by transmission electron microscope and Western blotting. (2) Exosomes were labeled with the green lipophilic fluorescent dye PKH67 and cultured with THP-1 macrophage to investigate whether HK2-derived exosomes could be internalized by THP-1 macrophage. Observing the morphology microscopically and detecting the chemotaxis function of THP-1 macrophages in Transwell chamber after co-cultured with exosomes. The expression of inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), Interleukin-1β (IL-1β), monocyte chemoattractant protein-1 (MCP-1) in cells and supernatants were separately detected by quantitative Real-time PCR (qRT-PCR) and enzyme linked immunosorbent assay (ELISA), and the expression of p-c-Jun NH2-terminal kinase (p-JNK), mitogen-activated protein kinase p-p38 (p-p38MAPK) and nuclear factor κB p65 (NF-κB p65) in THP-1 macrophages were detected by Western blot. Results (1) Vesicles that harvested by ultracentrifugation ranged in size from 30 nm to 100 nm and expressed exosomal marker CD63, TSG101 but absence of calnexin which is a marker of endoplasmic reticulum, suggesting that the exosomes were not contaminated with cells. (2) Results from laser scanning confocal microscope showed that each group of exosomes can be internalized by THP-1 macrophages. Compared with normal glucose exosomes group, high glucose exosomes had increased the expression of iNOS, TNF-α, IL-1β and MCP-1 in THP-1 macrophages (all P＜0.01), moreover, p-JNK, p-p38 MAPK and NF-κB p65 proteins level also increased significantly (all P＜0.01). Conclusions Exosomes from high glucose-treated HK2 cells can induce THP-1 macrophage activation and functional changes through MAPK/NF-κB pathway.

Objective To investigate effects of pirfenidone (PFD) on diabetic nephropathy model in db/db mice and to explore its possible mechanisms. Methods (1) Wild-type mice were as the normal control group, and db/db mice were divided into model group and PFD group, with 6 mice in each group. In the PFD group mice were administered continuously by 250 mg?kg-1?d-1 PFD for 18 weeks, and mice in the other two groups were administered by 0.5% sodium carboxymethyl cellulose. Blood glucose and 24 h urinary albumin were measured. The pathological changes of renal tissue were evaluated by PAS staining, PASM staining, Masson staining and Sirius red staining. The expression of collagen type Ⅳ in kidney tissues was detected by immunohistochemistry. (2) Mouse mesangial cells (SV40 MES-13 cells) were cultured as research objects. They were divided into control group, hyperosmolar group, high glucose (HG) group, and 50, 100, 200, 400, 800, 1600 mg/L PFD+HG group. BrdU cell proliferation test was used to evaluate cell proliferation rate. Cells were divided into control group, hyperosmolar group, HG group and PFD+HG group. The mRNA expressions of α-smooth muscle actin (α-SMA), collagen type Ⅰ, collagen type Ⅳ, transforming growth factor-β1 (TGF-β1), interleukin (IL)-1β, IL-6 and monocyte chemotactic protein-1 (MCP-1) were detected by real-time PCR. Results (1) Compared with normal control group, the model mice had higher weight, blood glucose and 24 h urinary albumin, accompanied with glomerular hypertrophy, mesangial area expansion, tubulointerstitial fibrosis and deposition of collagen type Ⅳ (all P＜0.05). Compared with those in model group, in PFD group 24 h urinary albumin decreased, glomerular hypertrophy, mesangial area expansion and tubulointerstitial fibrosis alleviated, and the protein expression of collagen type Ⅳ inhibited (all P＜0.05). (2) Compared with those in HG group, MES-13 cell proliferation rates of 100, 200, 400, 800, 1600 mg/L PFD+HG groups decreased (all P＜0.05), and the mRNA expressions of α-SMA, collagen type Ⅰ, collagen type Ⅳ, TGF-β1, IL-1β, IL-6 and MCP-1 reduced in 400 mg/L PFD+HG group (all P＜0.05). Conclusions PFD can inhibit high glucose-induced proliferation and activation of glomerular mesangial cells, decrease the expression of TGF-β1 and proinflammatory factors, as well as reduce the synthesis of collagen, which improve renal fibrosis of db/db mice.