Objective To investigate the effects of rituximab on lymphocytes and immunoglobulin in the treatment of focal segmental glomerulosclerosis (FSGS) and minimal change disease (MCD). Methods The subjects were FSGS and MCD patients admitted to Ruijin Hospital affiliated to Shanghai Jiaotong University on July 1, 2014 and July 1, 2019. All the enrolled patients were confirmed by clinical examination and renal biopsy, and received rituximab treatment (4 infusions of 375 mg/m² with the interval of 7-14 d). The levels of immunoglobulin IgA, IgG, IgM, and lymphocytes of CD19⁺, CD20⁺, CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺ and natural killer cells (CD56⁺CD16⁺) were compared between baseline and the third month, the sixth month, the ninth month and the twelfth month after treatment. Results Ninety-six patients with FSGS or MCD were enrolled in this study. The midian age was 28 years old (14-77 years old). The ratio of men to woman was 1.8∶1. There were 65 cases of MCD and 31 cases of FSGS. After rituximab treatment, the 24 h-proteinuria was significantly lower than that before treatment, and the serum albumin level was increased (both P<0.05). After rituximab treatment of 3 months, 6 months, 9 months and 12 months, CD19⁺ and CD20⁺ lymphocyte counts were significantly decreased (all P<0.01), and gradually recovered after 6 months. Compared with baseline, at 3, 6, 9, 12 months after rituximab treatment, the level of blood IgG was significantly increased (P=0.004,<0.001,<0.001,<0.001, respectively), and the level of blood IgM was significantly decreased (P<0.001, =0.008, =0.005,<0.001, respectively) but the median level still within the normal range (400-3 450 mg/L). The level of blood IgA was not significantly changed (all P<0.05). T lymphocytes (CD3⁺, CD3⁺CD4⁺ and CD3⁺CD8⁺) and natural killer cells (CD56⁺CD16⁺) showed no significant difference from baseline (all P>0.05). Conclusions Rituximab can effectively eliminate CD19⁺ and CD20⁺ lymphocytes, and has little influence on peripheral blood lymphocyte count and immunoglobulin level except CD19⁺ and CD20⁺ lymphocytes. The standard administration of rituximab is safe for patients with FSGS and MCD.

Objective To investigate the clinicopathological characteristics and influencing factors of kidney prognosis in primary IgA nephropathy (IgAN) patients. Methods The data of primary IgAN patients diagnosed with renal biopsy in the First Affiliated Hospital of Anhui Medical University from January 2015 to September 2019 were retrospective analyzed. According to the level of baseline estimated glomerular filtration rate (eGFR) when performing renal biopsy, the patients were divided into group A[eGFR≥90 ml·min^-1·(1.73 m²)^-1], group B[eGFR 61-89 ml·min^-1·(1.73 m²)^-1] and group C[eGFR≤60 ml·min^-1·(1.73 m²)^-1]. The clinical and pathological data were collected and compared among the three groups. Kaplan-Meier method was conducted for renal results, whereas the Cox proportional-hazards regression model was exploited to analyze the influencing factors of kidney prognosis in IgAN patients. Results A total of 742 patients were included in the study, including 394 cases (53.1%) in group A, 203 cases (27.4%) in group B, and 145 cases (19.5%) in group C. There were 325 males (43.8%) and 417 females (56.2%). The median duration of renal biopsy was 6 (1, 24) months, and the median age was 36 years old (18-68 years old). As the baseline level of renal function decreased, the proportion of patients with nephrotic syndrome, hypertension, anemia and hyperuricemia and the levels of 24 h urinary protein, serum triglyceride and total cholesterol increased significantly (all P<0.05), while the proportion of gross hematuria episodes and the ratio of serum albumin to globulin significantly decreased (all P<0.05). For the aspect of pathological manifestations, the proportions of cell proliferation in capillaries (E1), segmental sclerosis or adhesion (S1), renal tubular atrophy or interstitial fibrosis (T1/2), globular sclerosis, renal arteriole wall thickening and vitreous degeneration, Lee's gradeⅣ andⅤ increased with the decrease of baseline renal function (all P<0.05). Kaplan-Meier analysis showed that the cumulative renal survival rate decreased with the decline of baseline renal function (Log-rank χ²=88.510, P<0.001). As a result of multivariate Cox regression analysis, nephrotic syndrome (HR=2.399, 95%CI 1.054-5.459, P=0.037), hypertension (HR=1.806, 95%CI 1.071-3.048, P=0.027), low baseline eGFR (taking group A as the reference, group B: HR=2.383, 95%CI 1.053-5.392, P=0.037; group C: HR=6.878, 95%CI 3.074-15.393, P<0.001), IgG deposition (HR=2.224, 95%CI 1.384-3.574, P=0.001) and globular sclerosis (HR=2.075, 95%CI 1.230-3.501, P=0.006) were the independent influencing factors for renal progression in primary IgAN patients. Conclusions The level of baseline renal function in primary IgAN patients can be used to predict the extent of clinic-pathological damage. Nephrotic syndrome, hypertension, low baseline eGFR, IgG deposition and globular sclerosis are the independent influencing factors for renal progression in primary IgAN patients.

Objective To evaluate the value of combined measurement of urinary insulin-like growth factor-binding protein 7 (IGFBP7) and urinary metalloproteinase inhibitor -2 (TIMP-2) in the early diagnosis and prognosis of cardiac surgery-associated acute kidney injury (CSA-AKI). Methods From March 2018 to June 2018, cardiac surgery patients admitted to the cardiac macrovascular surgery department of the First Affiliated Hospital of Nanjing Medical University were prospectively included, and the blood creatinine was monitored to observe the presence of acute kidney injury (AKI). The prognostic information of the patients was collected, including in-hospital dialysis, in-hospital death, complete recovery of kidney function at discharge, death in one year after surgery, and progression to chronic kidney disease. The levels of urine IGFBP7 and TIMP-2 at 6 h, 24 h and 48 h after cardiac surgery were detected by enzyme linked immunosorbent assay (ELISA), and the urine creatinine (Cr) was also measured. Moreover, receiver operating characteristic curves (ROC) were plotted and the areas under the curves (AUC) were calculated to evaluate the predictive value and prognostic value of urinary [TIMP-2]·[IGFBP7] (T*I for short) and urine T*I/urine Cr² in CSA-AKI. Results A total of 74 patients with age of (58.43±10.91) years old and 47 males, were enrolled in this study, of which 24 cases (32.4%) had AKI and 10 cases (13.5%) had stage 2-3 AKI. Compared with the non-AKI group, the AKI group had significantly higher levels of urine T*I levels at 6 h and 24 h (both P<0.05). The AUC of T*I at 24 h predicting for AKI was 0.71(95%CI 0.59-0.81, P=0.001, cutoff value 0.020, sensitivity 79.2%, specificity 56.0%), while the AUC for stage 2-3 AKI was 0.85 (95%CI 0.75-0.92, P<0.001, cutoff value 0.083, sensitivity 70.0%, specificity 90.6%). Urinary T*I normalized for urinary creatinine excretion did not show better predictive value. In addition, of T*I at 24 h predicting for poor hospitalization outcome, renal recovery, and one year postoperative death, the AUC was 0.82(95%CI 0.71-0.90, P=0.001), 0.80(95%CI 0.66-0.86, P<0.001), and 0.81(95%CI 0.70-0.89, P=0.047), respectively. Conclusion The combined detection of TIMP-2 and IGFBP7 in urine is expected to be a biomarker for early diagnosis of CSA-AKI and has certain clinical value in predicting the prognosis of CSA-AKI.

Objective To explore the relationship between end-dialysis over-weight (edOW) in initial stage of hemodialysis and long-term prognosis in maintenance hemodialysis patients. Methods The data of initial uremia patients receiving hemodialysis in the First Affiliated Hospital, College of Medicine, Zhejiang University from January 1, 2008 to April 30, 2017 were retrospectively analyzed. The end point of follow-up was death or until April 30, 2018. The general data including age, gender, body mass index, primary disease, complications and laboratory indicators of the patients and the related parameters of dialysis from four to twelve months were collected. Kaplan-Meier method was used to analyze survival rate. Cox multivariate regression was used to analyze the relationship between edOW and all-cause mortality and cardiovascular disease (CVD) mortality. Results A total of 469 patients (300 males, 64.0%) were enrolled, with age of (56.9±17.1)years old. During the follow-up period of (4.1±2.4) years (1.0-10.3 years), 102 patients died. The main cause of death was cardiovascular and cerebrovascular events, accounting for 44.1%(45/102). The value of edOW was (0.28±0.02) kg. The patients were divided into edOW<0.28 kg group (n=292) and edOW≥0.28 kg group (n=177) according to the mean value of edOW. Kaplan-Meier survival analysis showed that the long-term survival rate in edOW<0.28 kg group was higher than that in edOW≥0.28 kg group (Log-rank χ²=4.134, P=0.043), and the CVD mortality in edOW≥0.28 kg group was significantly higher than that in edOW<0.28 kg group (Log-rank χ²=11.136, P=0.001). Cox multivariate regression analysis showed that high edOW was an independent influencing factor for all-cause death and CVD death in hemodialysis patients (HR=1.541, 95%CI 1.057-2.249, P=0.025; HR=1.930, 95%CI 1.198-3.107, P=0.007). Conclusion High edOW in early phase is an independent influencing factor of all-cause and CVD death in hemodialysis patients.

Objective To explore the relationship between metabolic acidosis and cardiac valve calcification in maintenance hemodialysis (MHD) patients in the Pearl River Delta Region. Methods Patients on MHD greater than 3 months who were treated in 10 blood purification centers in the Pearl River Delta Region from July 1 to September 30, 2019 were selected for this multicenter cross-sectional study. Based on a Doppler ultrasound, MHD patients were further divided into non-valve calcification group and valve calcification group. The demographics data, frequency of dialysis, blood pressure, single pool Kt/V(spKt/V), dialysis medications and laboratory data were collected and compared. Spearman correlation analysis was used to analyze the correlation between serum carbon dioxide combining power (CO₂CP) and cardiac valve calcification. Multivariate logistic regression model was used to analyze the influencing factors of cardiac valve calcification. Results A total of 664 MHD patients were included in this study, with age of (57.0±14.2) years old and dialysis age of 43.0(22.3, 71.7) months, including 395 males (59.5%) and 269 females (40.5%). Among them, there were 119 patients (17.9%) with diabetes and 186 patients (28.0%) with dialysis 2 times per week. There were 329 patients (49.5%) in the valve calcification group, and 335 patients (50.5%) in the non-valve calcification group. Compared to those in non-valve calcification group, valve calcification group had longer duration of dialysis, higher proportion of patients with dialysis 2 times per week, higher levels of diastolic blood pressure, fasting blood glucose, intact parathyroid hormone and ferritin, higher proportion of patients with blood CO₂CP<19 mmol/L (median CO₂CP), higher proportion of patients on usage of calcium channel blocker, angiotensin converting enzyme inhibitor/angiotensin receptor blocker, α-receptor blocker, β-receptor blocker, calcitriol and lanthanum carbonate (all P<0.05), while the levels of spKt/V, hemoglobin, serum CO₂CP, corrected calcium, blood phosphorus, blood alkaline phosphatase, albumin, total cholesterol, triacylglycerol, low-density lipoprotein, high-density lipoprotein, transferrin saturation, and the proportion of patients on usage of sevelamer and cinacalcet were lower (all P<0.05). Spearman analysis showed significant negative correlation between serum CO₂CP and valve calcification (r_s=-0.697, P<0.001). Multivariate logistic regression analysis showed that dialysis performed twice a week (OR=2.789, 95%CI 1.232-6.305, P=0.014), blood total cholesterol (OR=1.449, 95%CI 1.014-2.071, P=0.042), CO₂CP<19 mmol/L (OR=22.412, 95%CI 10.640-47.210, P<0.001) were the influencing factor of valve calcification in MHD patients. Conclusions MHD patients with cardiac valve calcification have significant acid loading. Metabolic acidosis is an independent influencing factor for cardiac valve calcification in MHD patients.

Objective To investigate the level of trimethylamine N-oxide (TMAO), one of gut metabolites, in patients undergoing maintenance hemodialysis (MHD) accompanied by congestive heart failure (HF) and its influencing factors. Methods Those patients of 18-75 years old who received three or more times of hemodialysis sessions per week for three months or longer during Nov 2018 and Mar 2019 were enrolled. Those attended health checkup at the same time without obvious kidney abnormality served as non-kidney disease controls. Serum TMAO concentrations were measured using high-performance liquid chromatography electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS). The levels of TMAO were compared between patients on hemodialysis and controls, between those with heart failure and without heart failure using logrithmically transformed TMAO (lnTMAO). Linear regression analysis was performed to investigate factors influencing TMAO levels. Results A total of 195 patients undergoing MHD and 40 controls were enrolled. Among them, 30 hemodialysis cases (15.4%) manifested as heart failure symptoms and /or left ventricular ejection fraction less than 50%. Males accounted for 67.2% in patients on hemodialysis and 37.5% in controls (χ²=12.426, P<0.001) respectively, while the median ages in both groups were 62.0(48.0, 71.0), 45.0(33.3, 55.0) years old respectively (Z=5.685, P<0.001). TMAO concentrations were significantly higher in patients on hemodialysis than controls [5.54(3.84, 8.91) mg/L vs 0.17(0.11, 0.30) mg/L, after log transformed, t=21.687, P<0.001]. However, there was no statistically significant difference between those with heat failure and those without in male [63.3% vs 67.9%, χ² =0.238, P=0.626], age [64.5(56.8, 71.0) years old vs 61.0(47.0, 72.0) years old, Z=0.894, P=0.372] and TMAO [5.17(3.30, 9.46) mg/L vs 5.57(3.87, 8.95) mg/L, after log transformed, t=-1.537, P=0.135]. Multivariate linear regression analysis demonstrated that in all the participants, serum urea was the main risk factor for TMAO [standardized coefficient (SB)=0.483]. lnTMAO=0.078×[serum urea(mmol/L)]+0.001×[serum creatinine (μmol/L)]-0.002×[serum uric acid (μmol/L)]-0.003×[platelet (×10⁹/L)]+0.014×[age (years old)]+0.344 (if diabetic)-1.266. While in those undergoing MHD, ultrafiltration volume had the most significant effect on TMAO levels (SB=0.279). lnTMAO=0.249×[ultrafiltration volume(L)]+0.059×[serum albumin (g/L)]+0.008×[age (years old)-0.526 (if heart failure existed)-1.865. Conclusions MHD patients have gut dysbiosis, while those hemodialysis patients accompanied by heart failure may have peculiar gut microbiota which induces lower serum TMAO levels than those without heart failure after adjusting for multiple related factors. Serum TMAO levels may be associated with ultrafiltration volume and nutrition status etc.

Objective To identify the differentially expressed genes and pathways of minimal change disease (MCD) by bioinformatics analysis, and to explore the pathogenesis of MCD. Methods The gene expression omnibus (GEO) under the National Center for Biotechnology Information (NCBI) platform of the United States was used, and the data chips GSE104948 and GSE104954 containing MCD information were selected. The data set contained the gene expression array data of 19 cases of MCD renal biopsy tissue and 36 cases of normal renal tissue. The online tool GEO2R was used to analyze data and screen differentially expressed genes, and DAVID 6.8 database was used to perform GO and KEGG functional enrichment analysis of differentially expressed genes and network analysis of genes involved in metabolic pathways. The String 11.0 database and Cytoscape 3.7.2 software were used to analyze the relationship between MCD differentially expressed genes and perform visual analysis. At the same time, the CytoHubba plug-in was used to analyze the degree of association of protein interaction networks and screen key expressed genes. Results A total of 302 highly expressed differentially expressed genes were identified by online tool GEO2R. GO analysis showed that the products of these differential genes were mostly located in the extracellular matrix, exosomes, pernucleus and other regions, exerting cell adhesion molecule binding, deoxycytidine deaminase activity, protein homodimerization activity, 2'-5'-oligoadenylic acid synthase activity and other functions, as well as participating in the formation of extracellular matrix, cell lysis, cell apoptosis, inflammatory response, immune response and other biological processes. KEGG analysis showed that differentially expressed genes were enriched in local adhesion, NOD-like receptor and other signal pathways. Combining the results of GO analysis and Cyto Hubba analysis, the PYCARD gene was screened out as the key gene that induced the inflammatory response in MCD kidney. Conclusions The inflammatory response may be involved in the occurrence and development of MCD, and the PYCARD gene may be a key gene in the induction of inflammatory response in MCD.

Objective To explore the effect of Wnt/β-catenin signaling pathway on growth plate development of tibial growth plate in young chronic renal failure (CRF) rats. Methods Four-week-old male SD rats were randomly divided into control group and CRF group (n=20/per group). Control group was intragastric administration with distilled water, and CRF group was given adenine suspension (150 mg·kg^-1·d^-1). All the young rats were sacrificed after continuous gavages for 6 weeks. The full length of tibia was compared between the two groups. The width of tibia proximal growth plates was measured by micro-CT scanning, and the width of the growth plate was also measured in histological sections. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation. Immunohistochemical staining was used to detect the expression of collagenⅡ, matrix metalloproteinase 13 (MMP-13) and β-catenin in chondrocytes. Western blotting was used to detect the protein expressions of collagenⅡ, MMP-13 and β-catenin. Results Compared with the control group, the tibial length of rats in the CRF group was shorter [(27.32±5.81) mm vs (35.43±3.61) mm, t=5.226, P<0.001]，the width of growth plate in micro-CT picture was more narrow [(0.72±0.22) mm vs (1.13±0.27) mm, t=5.096, P<0.001], and the relative width of the growth plate was also more narrow (t=6.744, P<0.001) in histological sections. The results of immunohistochemistry and Western blotting showed the expressions of collagenⅡ in the CRF group decreased significantly (t=8.212, P<0.001), MMP-13 (t=13.091, P<0.001) and β-catenin (t=7.534, P<0.001) increased significantly compared the control group in chondrocytes. Conclusion The Wnt/β-catenin signaling pathway is highly expressed in the tibial growth plate of young rats with chronic renal failure, which leads to accelerated degeneration and differentiation of chondrocytes and a closure tendency of growth plate.