Introduction: The objective of this study was to develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) method to detect phospholipase A2 receptor (PLA2R)-IgG1 antibodies and evaluate its clinical relevance in predicting the prognosis of individuals with idiopathic membranous nephropathy (IMN). Materials and methods: A three-step indirect TRFIA method was established using a PLA2R antigen-coated microtiter plate to capture PLA2R-IgG antibodies, followed by detection using mouse anti-human IgG1 and Eu3+labeled goat anti-mouse IgG antibodies. This method was applied to the initial serum of 56 patients with PLA2RIMN to investigate the clinical value of PLA2R-IgG1 antibody levels in predicting IMN prognosis. Results: The detection range of PLA2R-IgG1-TRFIA was 0.85-300 RU/mL, with intra-assay precision of 3.54-5.93 % and inter-assay precision of 4.39-9.36 %. Recoveries were 101.77-108.04 %. A PLA2R-IgG1 level above 2.21 RU/mL indicated PLA2R-IMN. At initial diagnosis, the median PLA2R-IgG level was 51.24 RU/mL in the remission group and 93.27 RU/mL in the non-remission group. The median PLA2R-IgG1 level was 603.32 RU/mL in the non-remission group, which was 4.29 times higher than that in the remission group (140.67 RU/mL). PLA2R-IgG1 levels (P = 0.001) more effectively distinguished between remission and nonremission groups compared with PLA2R-IgG levels (P = 0.094). Conclusions: The first quantitative TRFIA for PLA2R-IgG1 was established, showing greater clinical value in predicting IMN prognosis, compared to that for PLA2R-IgG levels.

Autophagy, metabolism, and associated signaling pathways play critical roles in bacterial survival within mammalian cells and influence the immunopathogenesis of infections. Mesenchymal stem cells (MSCs) are important host cells during Mycobacterium tuberculosis (Mtb) infection, yet how autophagy, metabolism, and related pathways are modulated in MSCs infected with the virulent H37Rv or the attenuated H37Ra strain of Mtb remains poorly understood. In this study, we utilized RNA-Seq screening, qRT-PCR, and Western Blotting to investigate the differences in these processes between H37Rv and H37Ra infections. Our results show that, at early time points (no more than 24h), infection with H37Rv significantly increased the expression of TlLR2, Prkaa2, and Prkaa2 phosphorylation in MSCs compared with H37Ra infection. Further analysis revealed that H37Rv infection induced a stronger autophagic response (evidenced by increased Atg9b and LC3II/LC3I) through the TLR2-AMP-AMPK pathway than H37Ra infection. Despite these differences in autophagy, there was no statistically significant difference in bacillary loads, suggesting that, in addition to autophagy, other factors such as apoptosis and immune-inflammatory responses may also regulate Mtb growth in MSCs. Additionally, the metabolic analysis showed that H37Rv infection led to increased expression of SLC2A3, PFKFB3, HK1, and ABCA1 in MSCs compared to H37Ra infection. These findings confirm that, during the early stages of infection, H37Rv induces enhanced autophagy, glucose metabolism, and cholesterol efflux through a more active TLR2-AMP-AMPK pathway than H37Ra. Therefore, MSCs may represent a novel target for the prevention and treatment of tuberculosis.

Introduction: The role of dietary sodium intake in the risk of chronic kidney disease progression remains controversial. This study aimed to evaluate the association of urinary sodium excretion and progression of IgA nephropathy. Methods: We assessed 596 patients with IgA nephropathy, and urinary sodium excretion was measured at the time of kidney biopsy. Cox proportional hazards models and restricted cubic splines were used to assess the association between urinary sodium excretion and kidney disease progression events, defined as 50% eGFR decline or development of kidney failure. Results: After a mean follow-up of 58.9 months, a total of 75 (12.6%) participants of IgA nephropathy reached composite kidney disease progression events. The risk of kidney disease progression events was higher in patients with higher urinary sodium excretion. After adjustment for traditional risk factors, higher levels of ln-transformed urinary sodium excretion was associated with the kidney disease progression events in patients with IgA nephropathy (HR: 2.1; 95% CI: 1.4-3.2). In reference to the fi rst tertile of urinary sodium excretion, hazard ratios were 1.9 (95% CI: 1.0-3.4) for the second tertile and 2.1 (95% CI: 1.1-3.9) for the third tertile. Conclusion: Higher levels of urinary sodium excretion were associated with kidney disease progression events in IgA nephropathy independent of clinical and biopsy characteristics.

BACKGROUND:Minimal change disease (MCD) is a common pathological type of nephrotic syndrome. Relapses of MCD present a significant challenge for patients. This study aims to develop a predictive model for evaluating the probability of relapse in patients with MCD. METHODS:This study enrolled 152 patients with biopsy-confirmed MCD, all of whom received exclusive glucocorticoid treatment at the First Affiliated Hospital of Zhejiang University in Hangzhou, China, between October 2012 and April 2021. The Cox regression analysis was utilized to identify the risk factors associated with the relapse in MCD, and a nomogram was constructed to predict the probability of relapse. RESULTS:The results demonstrated that serum immunoglobulin E (IgE) levels > 936 IU/mL, age ≤ 30 years old, estimated glomerular filtration rate (eGFR) < 90 mL/(min × 1.73 m2), serum total cholesterol (TCh) levels > 12.3 mmol/L, and time to remission were independent risk factors associated with relapses in patients with MCD. A nomogram was established and achieved a concordance index of 0.726 (95% CI = 0.659-0.793). The receiver operating characteristic curves demonstrated areas under the curve of 0.771, 0.853, and 0.811 for the prediction of relapse at 1-, 2-, and 3-year intervals after achieving remission in MCD patients respectively. These findings along with the calibration curves indicated the good discrimination and calibration performance of the nomogram in this cohort. The established nomogram provides a valuable tool for the identification of patients with MCD who are at risk of relapse, which may facilitate prompt treatment for these patients.

BACKGROUND:The analytical renal pathology system (ARPS) based on convolutional neural networks has been used successfully in native IgA nephropathy (IgAN) patients. Considering the similarity of pathologic features, we aim to evaluate the performance of the ARPS in allograft IgAN patients and broaden its implementation.METHODS:Biopsy-proven allograft IgAN patients from two different centers were enrolled for internal and external validation. We implemented the ARPS to identify glomerular lesions and intrinsic glomerular cells, and then evaluated its performance. Consistency between the ARPS and pathologists was assessed using intraclass correlation coefficients. The association of digital pathological features with clinical and pathological data was measured. Kaplan-Meier survival curve and cox proportional hazards model were applied to investigate prognosis prediction.RESULTS:A total of 56 biopsy-proven allograft IgAN patients from the internal center and 17 biopsy-proven allograft IgAN patients from the external center were enrolled in this study. The ARPS was successfully applied to identify the glomerular lesions (F1-score, 0.696-0.959) and quantify intrinsic glomerular cells (F1-score, 0.888-0.968) in allograft IgAN patients rapidly and precisely. Furthermore, the mesangial hypercellularity score was positively correlated with all mesangial metrics provided by ARPS [Spearman's correlation coefficient (r), 0.439-0.472, and all p values < 0.001]. Besides, a higher allograft survival was noticed among patients in the high-level groups of the maximum and ratio of endothelial cells, as well as the maximum and density of podocytes.CONCLUSION:We propose that the ARPS could be implemented in future clinical practice with outstanding capability.

Background: The Gitelman syndrome (GS) is an autosomal recessive disorder of renal tubular salt handling. Gitelman syndrome is characterized by hypokalemia, metabolic alkalosis, hypomagnesemia, hypocalciuria, and renin-angiotensin-aldosterone system (RAAS) activation, and is caused by variants in the SLC12A3 gene. Gitelman syndrome has a heterogeneous phenotype, which may or may not include a range of clinical signs, posing certain difficulties for clinical diagnosis.Case presentation: A 49-year-old man was admitted to our hospital due to muscular weakness. The patient’s history revealed previous recurrent muscular weakness events associated with hypokalemia, featured by a minimum serum potassium value of 2.3 mmol/L. The reported male patient had persistent hypokalemia, hypocalciuria and normal blood pressure, without presenting obvious metabolic alkalosis, growth retardation, hypomagnesemia, hypochloremia or RAAS activation. We performed whole-exome sequencing and identified a novel compound heterozygous variant in the SLC12A3 gene, c.965-1_976delGCGGACATTTTTGinsACCGAAAATTTT in exon8 and c.1112T>C in exon9 in the proband.Conclusion: This is a study to report a heterogeneous phenotype Gitelman syndrome with a novel pathogenic compound heterozygous variant in the SLC12A3 gene. This genetic study expands the variants spectrum, and improve the diagnostic accuracy of Gitelman syndrome. Meanwhile, further functional studies are required to investigate the pathophysiological mechanisms of Gitelman syndrome.

INTRODUCTION:Coagulation disorders play a key role in chronic kidney disease, and the formation or elevation of plasma D-dimer levels reflects activation of the coagulation system. However, its relationship with the severity and progression of kidney disease in IgA nephropathy (IgAN) remains unclear. METHODS:We assessed 1818 patients with IgAN diagnosed between 2002 and 2019 at the First Affiliated Hospital, Zhejiang University School of Medicine. Plasma D-dimer levels were measured at the time of the renal biopsy. The association between plasma D-dimer levels and kidney disease progression events, defined as a 50% decline in eGFR and end-stage kidney disease (ESKD), was tested using restricted cubic splines and Cox proportional hazard models. RESULTS:The median plasma D-dimer level was 220 (170-388.5) µg/L FEU, which was significantly higher than healthy controls 170 (170-202) µg/L FEU. Plasma D-dimer levels were positively correlated with proteinuria (r = 0.211, p < 0.001) and serum galactose-deficient IgA1 (r = 0.226, p = 0.004) and negatively correlated with eGFR (r=-0.127, p < 0.001) and Oxford T (p < 0.001) and C (p = 0.004) scores. After a median follow-up of 25.67 (13.03-47.44) months, 126 (6.93%) patients experienced composite kidney disease progression events. Higher plasma D-dimer levels were associated with an increased risk of kidney disease progression events (hazard ratio, 1.73; 95% confidence interval [95% CI], 1.40-2.23) per ln-transformed plasma D-dimer (p < 0.001), after adjustment for sex, age, proteinuria, Mean arterial pressure (MAP) and Oxford classification scores. In reference to the first tertile of plasma D-dimer, hazard ratios were 1.48 (95% CI, 0.76-2.88) for the second tertile, 3.03 (95% CI, 1.58-5.82) for the third tertile. CONCLUSIONS:High plasma D-dimer levels were associated with the progression of kidney disease severity in IgA nephropathy.

Abstract Background and Aims Anti-CD20 monoclonal antibody has become one of the first-line therapies for the treatment of moderate and high-risk primary membranous nephropathy (pMN). A novel glycoengineered type Ⅱ anti-CD20 antibody, MIL62 with a nearly completely afucosylated N-glycans in Fc region, has demonstrated superior activity compared with rituximab and obinutuzumab in vitro and in vivo, respectively. To evaluate the safety, tolerability, pharmacokinetics, pharmacodynamics and efficacy of MIL62 in pMN, we conducted a multicenter, randomized, controlled, open-label, phase Ib/Ⅱ study (NCT05398653). Method This study aims to investigated MIL62 or Cyclosporine for the treatment of Chinese pMN. Eligible patients with pMN diagnosed by kidney biopsy, proteinuria of at least 3.5 g per 24 hours received intravenous MIL62 (two infusions, 600 or 1000 mg each, administered 14 days apart; repeated at 6 months) or Cyclosporine (starting at a dose of 3.5 mg per kilogram of body weight per day for 12 months). Patients were followed up for up to 104 weeks. The primary outcome was a composite of complete or partial remission of proteinuria and stable eGFR at 76 weeks. Laboratory variables and safety were also assessed. Results From February 23th, 2022 to December 21th, 2022, 85 patients were randomly enrolled from 19 centers in China, and received at least one dose of MIL62 or Cyclosporine. At the data cut-off date (January 30th, 2023), 60 patients were followed up for at least 12 weeks, 24/40 (60%) patients in the MIL62 group and 7/20 (35%) patients in the Cyclosporine group achieved complete or partial remission; 26 patients were followed up for at least 24 weeks, 11/18 (61.1%) patients in the MIL62 group and 2/8 (25%) patients in the Cyclosporine group achieved partial remission. In the 56 patients who were positive for anti-PLA2R antibodies (≥14RU/mL) at baseline and followed up for at least 12 weeks, 29/38 (76.3%) patients in the MIL62 group achieved immunological remission (PLA2R Abs<14RU/mL), which was superior to the Cyclosporine group where 8/18 (44.4%) patients achieved immunological remission (P< 0.05). The remission to MIL62 in our study was rapid because 61.1% of patients achieved complete or partial remission at 24 weeks compared with the 35% 6-month response rate reported in the Membranous Nephropathy Trial of Rituximab study [1]. Among the 85 safety-evaluable patients, treatment emergent adverse events (TEAEs) occurred in 23/30 (76.7%) patients in the MIL62 600 mg group, 26/30 (86.7%) patients in the MIL62 1000 mg group and 24/25 (96.0%) patients in the Cyclosporine group; treatment-related AEs (TRAEs) occurred in 21/30 (70.0%), 23/30 (76.7%) and 18/25 (72.0%) in the MIL62 600 mg, MIL62 1000 mg and Cyclosporine group respectively; Grade 3 or above TEAEs were observed in 5 (16.7%), 1 (3.3%) and 2 (8.0%) patients in the MIL62 600 mg, MIL62 1000 mg and Cyclosporine group respectively; Serious adverse events (SAEs) occurred in 5 (16.7%), 0 (0.0%) and 2 (8.0%) patients in the MIL62 600 mg, MIL62 1000 mg and Cyclosporine group respectively. Only one patient in the MIL62 600 mg group experienced serious treatment-related thrombocytopenia and has recovered so far; Other SAEs are not related to treatment. B cell depletion occurred within 24 hours after MIL62 infusion and could last for 24 weeks. Conclusion The 12-week immunological remission rate in the MIL62 group was significantly higher than the Cyclosporine group. MIL62 had a manageable safety profile. This phase Ib/II preliminary data warrant further phase III clinical trials of MIL62 in pMN.

Inflammatory pathways involving Mesenchymal stromal cells (MSCs) play an important role in Mycobacterium tuberculosis (Mtb) infection. H37Rv (Rv) is a standard virulent strain, however, H37Ra (Ra) is a strain with reduced virulence. Interleukins and chemokines production are known to promote inflammation resistance in mammalian cells and is recently reported to regulate mycobacterial immunopathogenesis via inflammatory responses. MSCs are very important cells during Mtb infection. However, the different expressions of interleukins and chemokines in the process of Mtb-infected MSCs between Ra and Rv remain unclear. We used the techniques of RNA-Seq, qRT-PCR, ELISA, and Western Blotting. We have shown that Rv infection significantly increased mRNA expressions of Mndal, Gdap10, Bmp2, and Lif, thereby increasing more differentiation of MSCs compared with Ra infection in MSCs. Further investigation into the possible mechanisms, we found that Rv infection enhanced more inflammatory response (Mmp10, Mmp3, and Ptgs2) through more activation of the TLR2-MAP3K1-JNK pathway than did Ra infection in MSCs. Further action showed that Rv infection enhanced more Il1α, Il6, Il33, Cxcl2, Ccl3, and Ackr3 production than did Ra infection. Rv infection showed more expressions of Mmp10, Mmp3, Ptgs2, Il1α, Il6, Il33, Cxcl2, Ccl3, and Ackr3 possibly through more active TLR2-MAP3K1-JNK pathway than did Ra infection in MSCs. MSCs may therefore be a new candidate for the prevention and treatment of tuberculosis.

BackgroundProteinuria is a strong risk factor for renal outcomes in IgA nephropathy. Random urine protein-to-creatinine ratio (PCR), random albumin-to-creatinine ratio (ACR), and 24-h urine protein excretion (24-h UP) have been widely used in clinical practice. However, the measurement which is the best predictor of long-term renal outcomes remains controversial. This study aimed to compare the three measurements in IgA nephropathy.MethodsWe conducted a retrospective study of 766 patients with IgA nephropathy. The associations among baseline ACR, PCR, and 24-h UP with chronic kidney disease (CKD) progression event, defined as 50% estimated glomerular filtration rate (eGFR) decline or end stage kidney disease (ESKD), were tested and compared.ResultsIn this study, ACR, PCR, and 24-h UP showed high correlation (r = 0.671–0.847, P < 0.001). After a median follow-up of 29.88 (14.65–51.65) months, 51 (6.66%) patients reached the CKD progression event. In univariate analysis, ACR performed better in predicting the prognosis of IgA nephropathy, with a higher area under the receiver operating curve (ROC) curve than PCR and 24-h UP. After adjustment for traditional risk factors, ACR was most associated with composite CKD progression event [per log-transformed ACR, hazard ratio (HR): 2.82; 95% (95% CI): 1.31–6.08; P = 0.008].ConclusionsIn IgA nephropathy, ACR, PCR, and 24-h UP had a high correlation. ACR performed better in predicting the prognosis of IgA nephropathy.

BackgroundHypertension has been shown to be an important risk factor in IgA nephropathy (IgAN). The 2021 the Kidney Disease Improving Global Outcomes (KDIGO) Guideline proposes a target systolic blood pressure (SBP) of less than 120 mmHg in patients with Chronic Kidney Disease (CKD) not receiving dialysis. However, whether lowering SBP from <140– <120 mm Hg is renoprotective is unknown. This study aims to evaluate the association of SBP and the progression of IgAN, then explore whether lowering SBP from <140– <120 mm Hg is renoprotective.MethodsOverall, 2,240 patients with IgAN were enrolled in this study. Cox proportional hazards models and restricted cubic splines were used to estimate the associations between SBP and kidney failure events which are defined as 50% estimated glomerular filtration rate (eGFR) decline or kidney failure.ResultsAfter a median follow-up of 30.05 months, 217 (9.69%) patients reached composite kidney failure events. The association of SBP and kidney failure events showed a linear relationship. The risk of kidney failure events was greater with higher SBP. Compared with SBP <120 mm Hg, the hazard ratio was 1.85 (1.16–2.97, p = 0.010) for SBP <140 mm Hg after adjustment for traditional risk factors. The renoprotective benefits of therapy targeting SBP <120 mm Hg from SBP <140 mm Hg was detectable within the subgroup with proteinuria >1.0 g/d, CKD 1-3a stage, but not those with proteinuria ≤ 1.0 g/d and CKD 3b-4 stage.ConclusionsIn patients with IgAN, SBP was independently associated with composite kidney failure events. Lowering SBP from <140– <120 mm Hg was renoprotective.

Background A large number of studies have shown that serum globulin plays an important role in a variety of cancers; However, few studies have identified the association between serum globulin levels and end-stage renal disease (ESRD) and all-cause death in Chinese patients with multiple myeloma (MM). Methods A generalized additive model and smooth curve fitting were fitted to assess the cross-sectional relationship between the serum globulin levels and renal impairment (RI) at baseline. Multivariate-adjusted Cox regression models were performed to determine the associations between the baseline serum globulin levels and the onset of all-cause death and ESRD in patients with MM. Results 288 participants who were followed for > 3 months were eligible for the retrospective study. The median serum globulin level was 5.1 ± 2.6 mg/dL. The average follow-up time was 23.3 months. Thirty-two patients (11.5%) had ESRD and 24 patients (8.33%) died after diagnosis. In patients with a serum globulin level < 6.1 mg/dL, the serum globulin level had an independent, negative correlation with the occurrence of MM-related RI. Patients were divided into three groups on the basis of serum globulin tertiles: low (L group), 3.3 mg/dL; middle (M group), 3.3–6.0 mg/dL; and high (H group), 6.0 mg/dL. Cox regression analysis showed that low serum globulin levels may be independent risk factors for all-cause death and the occurrence of ESRD in patients with MM; however, an elevated baseline serum globulin can predict all-cause deaths in patients with MM, but cannot predict the onset of ESRD. Conclusions This observational study suggested that there was a non-linear relationship between the serum globulin level and the occurrence of RI in patients with MM. This finding showed that the serum globulin level had a U-shaped association with all-cause death and an L-shaped association with ESRD in patients with MM.

[This corrects the article DOI: 10.1016/j.gendis.2020.07.005.].

BackgroundMinimal change disease (MCD) has a high recurrence rate, but currently, no biomarker can predict its recurrence. To this end, this study aimed at identifying potential serum cytokines as valuable biomarkers for predicting the risk of MCD recurrence.Materials and methodsRaybiotech 440 cytokine antibody microarray was used to detect the serum samples of eight relapsed, eight non-relapsed MCD patients after glucocorticoid treatment, and eight healthy controls. The differentially expressed cytokines were confirmed by enzyme-linked immunosorbent assay (ELISA) with serum samples from 29 non-relapsed and 35 relapsed MCD patients. The study used the receiver operating characteristic (ROC) curve analysis to investigate the sensitivity and specificity of a serum biomarker for predicting the MCD relapse.ResultsSerum IL-12p40 levels increased significantly in the relapsed group. The Area Under the ROC Curve (AUC) of IL-12p40 was 0.727 (95%CI: 0.597–0.856; P < 0.01). The RNA-sequencing analysis and qPCR assay performed on the IL-12 treated mouse podocytes and the control group showed increased expression of podocyte damage genes, such as connective tissue growth factor (CTGF), matrix metallopeptidase 9 (MMP9), secreted phosphoprotein 1 (SPP1), and cyclooxygenase-2 (COX-2) in the former group.ConclusionIL-12p40 may serve as a new biomarker for predicting the risk of MCD recurrence after glucocorticoid treatment, and it may be involved in the pathogenesis and recurrence of MCD.

Background Previous reports showed that some patients with minimal change disease (MCD) had high serum immunoglobulin E (IgE) levels. This study aimed to explore the proportion of MCD patients with high serum IgE levels and evaluate the correlation between serum IgE levels and MCD remission and relapse. Methods This study enrolled 222 new-onset patients with renal biopsy-confirmed MCD from October 2012 to October 2019 at the First Affiliated Hospital of Zhejiang University in Hangzhou, China. Patients’ demographics and clinical parameters were analyzed. Results The results indicated that 70.3% of 222 MCD patients had high serum IgE levels (IgE > 100.0 IU/mL). Moreover, 134 patients were treated with glucocorticoids alone and divided into the low- and high-IgE groups, according to the median serum IgE level (523.5 IU/mL). The mean time to complete remission of the low- and high-IgE groups was 29.0 ± 2.2 and 45.7 ± 4.2 days, respectively (log-rank test; P = 0.002). The mean time to total remission was 19.1 ± 1.4 and 31.6 ± 3.2 days of the low- and high-IgE groups, respectively (log-rank test; P < 0.001). The mean time to first relapse in the low- and high-IgE groups was 701.2 ± 65.0 and 425.0 ± 52.6 days, respectively (log-rank test; P = 0.002). Serum IgE ≥ 523.5 IU/mL was an independent correlation factor affecting the patients’ remission and relapse. Conclusion Serum IgE level was an independent correlation factor for MCD remission and relapse. MCD patients with high serum IgE levels were prone to delayed remissions and early relapses.

[This corrects the article DOI: 10.1016/j.gendis.2020.07.005.][This corrects the article DOI: 10.1016/j.gendis.2021.09.001.].