Presently, no specific therapies have been recognized for immunoglobulin A nephropathy (IgAN). Mycophenolate mofetil (MMF) has been verified effective for Chinese patients with IgAN. Telitacicept is a full-human TACI-FC fusion preventing B cells maturation and activation, and it has been proven to be beneficial for IgAN in a phase II clinical trial. This study was designed to observe the efficacy and safety of telitacicept plus low-dose MMF for IgAN treatment. This retrospective cohort study included 24 patients with IgAN, and patients were treated with telitacicept plus MMF. The primary outcome was settled as the changing in proteinuria and estimated glomerular filtration rate (eGFR). The subordinate outcome was set as the changing in hematuria. The mean follow-up time was 23 months. The median baseline proteinuria was 2.5 (1.74, 6.58) g/d, and eGFR was 94.97 (56.8, 120.67) mL/min/1.73 m2. There were noteworthy reductions in proteinuria at 3, 6, 9, 12, 15, 18, 21 and 24 months when compared to the baseline levels [1.45 (0.78, 1.8) g/d [p = 0.0122], 0.505 (0.26, 0.99) g/d [p < 0.0001], 0.48 (0.28, 0.76) g/d [p < 0.0001], 0.3 (0.17, 0.85) g/d [p < 0.0001], 0.23 (0.18, 0.575) g/d [p < 0.0001], 0.18 (0.12, 0.325) g/d [p < 0.0001], 0.14 (0.105, 0.22) g/d [p < 0.0001] and 0.14 (0.103, 0.278) g/d [p < 0.0001]]. All patients maintained stable eGFR during follow-up times. Besides, telitacicept plus MMF remarkably alleviated the hematuria. Telitacicept plus MMF treatment led to not only remarkable clinically significant reduction in proteinuria and hematuria, but also stable serum creatinine value of patients with IgAN without adverse side effects.

Renal tubular epithelial cells (TECs) undergo an energy-related metabolic shift from fatty acid oxidation to glycolysis during chronic kidney disease (CKD) progression. However, the mechanisms underlying this burst of glycolysis remain unclear. Herein, a new critical glycolysis regulator, the transcription factor forkhead box protein K1 (FOXK1) that is expressed in TECs during renal fibrosis and exhibits fibrogenic and metabolism-rewiring capacities is reported. Genetic modification of the Foxk1 locus in TECs alters glycolytic metabolism and fibrotic lesions. A surge in the expression of a set of glycolysis-related genes following FOXK1 protein activation contributes to the energy-related metabolic shift. Nuclear-translocated FOXK1 forms condensate through liquid-liquid phase separation (LLPS) to drive the transcription of target genes. Core intrinsically disordered regions within FOXK1 protein are mapped and validated. A therapeutic strategy is explored by targeting the Foxk1 locus in a murine model of CKD by the renal subcapsular injection of a recombinant adeno-associated virus 9 vector encoding Foxk1-short hairpin RNA. In summary, the mechanism of a FOXK1-mediated glycolytic burst in TECs, which involves the LLPS to enhance FOXK1 transcriptional activity is elucidated.

Abstract Objectives Pulmonary lesion is frequently seen in ANCA-associated vasculitis (AAV) patients primarily due to AAV lung involvement or infection, which are hard to differentiate due to their high similarity in clinical manifestations. We aimed to analyze the clinical features of pulmonary lesions consequent to AAV involvement or infection in AAV patients and further identify the markers for differential diagnosis. Methods 140 AAV patients who admitted to the Renmin Hospital of Wuhan University from January 2016 to July 2021 were included in this study. According to the nature of lung conditions, these patients were divided into the non-pulmonary lesion group, the lung infection group and the non-pulmonary infection group, and their demographics, clinical symptoms, imaging features, as well as laboratory findings were compared. A receiver operating characteristic (ROC) curve was drawn, and the diagnostic efficacy of single biomarker and composite biomarkers on pulmonary infection was then evaluated. Results The patients in the lung infection group were significantly older than those in the no lesion group (63.19 ± 14.55 vs 54.82 ± 15.08, p = 0.022). Patients in the lung infection group presented more frequent symptoms and more obvious pulmonary image findings. Compared with patients in the non-pulmonary infection group, patients in the lung infection group showed a higher symptom incidence of fever, chest tightness, cough and expectoration, and hemoptysis (52.94% vs 16.00%, 61.76% vs 40.00%, 72.06% vs 46.00%, 27.94% vs 8.00%, p < 0.05, respectively), and more changes in pulmonary CT scanning images in terms of patched/striped compact opacity, alveolar hemorrhage, bronchiectasis, pleural effusion, as well as mediastinal lymphadenopathy (89.71% vs 52.00%, 11.76% vs 2.00%, 22.06% vs 8.00%, 50.00% vs 20.00%, 48.53% vs 24.00%, p < 0.05, respectively). In addition, patients in the lung infection group had significantly higher levels of serum pro-calcitonin (PCT), C-reactive protein (CRP), amyloid A (SAA), blood neutrophil-to-lymphocyte ratio (NLCR), erythrocyte sedimentation rate (ESR), as well as Birmingham vasculitis activity score (BVAS) than patients in the other two groups (p < 0.05). Among all biomarkers, PCT exhibited the highest diagnostic efficacy (0.928; 95%CI 0.89–0.97) for pulmonary infected AAV patients at a cut-off score of 0.235 ng/ml with 85.3% sensitivity and 84% specificity. Moreover, the composite biomarker of PCT-CRP-NLCR showed more diagnostic efficacy (0.979; 95% CI 0.95–1.00) in distinguishing the infectious and non-infectious lung injuries in AAV patients. Conclusions AAV patients with lung infection manifested more clinical symptoms and prominent lung image changes. The PCT and composite biomarker PCT-CRP-NLCR showed high diagnostic efficacy for a lung infection in AAV patients. Pulmonary lesion caused by either infection or AAV involvement is commonly seen and difficult to distinguish. We aim to identify the biomarkers that can be applied in the differentiation diagnosis of pulmonary lesions in AAV patients.

Renal fibrosis is an unavoidable end result of all forms of progressive chronic kidney diseases (CKD). Discovery of efficacious drugs against renal fibrosis is in crucial need. In a preliminary study we found that a derivative of artemisinin, dihydroartemisinin (DHA), exerted strong renoprotection, and reversed renal fibrosis in adenine-induced CKD mouse model. In this study we investigated the anti-fibrotic mechanisms of DHA, particularly its specific target in renal cells. Renal fibrosis was induced in mice by unilateral ureteral obstruction (UUO) or oral administration of adenine (80 mg · kg −1 ), the mice received DHA (30 mg · kg −1 · d −1 , i.g.) for 14 or 21 days, respectively. We showed that DHA administration markedly attenuated the inflammation and fibrotic responses in the kidneys and significantly improved the renal function in both the renal fibrosis mouse models. In adenine-treated mice, DHA was more effective than 5-azacytidine against renal fibrosis. The anti-fibrotic effects of DHA were also observed in TGF-β1-treated HK-2 cells. In order to determine the target protein of DHA, we conducted pull-down technology coupled with shotgun proteomics using a small-molecule probe based on the structure of DHA (biotin-DHA). As a results, DNA methyltransferase 1 (DNMT1) was identified as the anti-fibrotic target of DHA in 3 different types of renal cell lines (HK-2, HEK293 and 3T3). We demonstrated that DHA directly bound to Asn 1529 and Thr 1528 of DNMT1 with a Kd value of 8.18 μM. In primary mouse renal tubular cells, we showed that DHA (10 μM) promoted DNMT1 degradation via the ubiquitin–proteasome pathway. DHA-reduced DNMT1 expression effectively reversed Klotho promoter hypermethylation, which led to the reversal of Klotho protein loss in the kidney of UUO mice. This subsequently resulted in inhibition of the Wnt/β-catenin and TGF-β/Smad signaling pathways and consequently conferred renoprotection in the animals. Knockdown of Klotho abolished the renoprotective effect of DHA in UUO mice. Our study reveals a novel pharmacological activity for DHA, i.e., renoprotection. DHA exhibits this effect by targeting DNMT1 to reverse Klotho repression. This study provides an evidence for the possible clinical application of DHA in the treatment of renal fibrosis.

Renal fibrosis is an inevitable outcome of various manifestations of progressive chronic kidney diseases (CKD). The need for efficacious treatment regimen against renal fibrosis can therefore not be overemphasized. Here we show a novel protective role of Bacteroides fragilis ( B. fragilis ) in renal fibrosis in mice. We demonstrate decreased abundance of B. fragilis in the feces of CKD patients and unilateral ureteral obstruction (UUO) mice. Oral administration of live B. fragilis attenuates renal fibrosis in UUO and adenine mice models. Increased lipopolysaccharide (LPS) levels are decreased after B. fragilis administration. Results of metabolomics and proteomics studies show decreased level of 1,5-anhydroglucitol (1,5-AG), a substrate of SGLT2, which increases after B. fragilis administration via enhancement of renal SGLT2 expression. 1,5-AG is an agonist of TGR5 that attenuates renal fibrosis by inhibiting oxidative stress and inflammation. Madecassoside, a natural product found via in vitro screening promotes B. fragilis growth and remarkably ameliorates renal fibrosis. Our findings reveal the ameliorative role of B. fragilis in renal fibrosis via decreasing LPS and increasing 1,5-AG levels.

Abstract Background Increasing evidence has indicated that circular RNAs (circRNAs) play a role in various diseases. However, the influence of circRNAs in nephritis remains unknown. Methods Microarray analysis and RT-qPCR were used to detect the expression of circRNA. Type I IFN were administrated to RMC and HEK293 cells to establish a nephritis cell model. CCK-8, MTT assay, and flow cytometry were used to assess cell proliferation, viability, and apoptosis of cells. Bioinformatics analysis and dual luciferase reporter assay detect the interaction of circ_0007059, miRNA-1278, and SHP-1. Glomerulonephritis was performed in a mouse model by administration of IFNα-expressing adenovirus. IHC staining showed the pathogenic changes. Results In the present study, the expression of circ_0007059 in type I interferon (IFN)-treated renal mesangial cells (RMCs), lupus nephritis (LN) specimens, and HEK293 cells was downregulated compared with that in normal healthy samples and untreated cells. Circ_0007059 overexpression resulted in increased cell proliferation, cell viability, apoptosis, and inflammation-associated factors (CXCL10, IFIT1, ISG15, and MX1) in RMCs and HEK293 cells. In addition, circ_0007059 overexpression significantly restored cell proliferation and viability and inhibited IFN-induced apoptosis. Further, the increased expression resulted in reduced inflammation and the downregulation of CXCL10, IFIT1, ISG15, and MX1 in RMCs and HEK293 cells. Circ_0007059 serves as a sponge for miR-1278 so that the latter can target the 3′-untranslated region of SHP-1. Overexpressed circ_0007059 inhibited miR-1278 expression and elevated SHP-1 expression, subsequently reducing STAT3 phosphorylation. Meanwhile, miR-1278 was upregulated and SHP-1 was downregulated in LN samples and IFN-treated cells. The restoration of miR-1278 counteracted the effect of circ_0007059 on viability, apoptosis, and inflammation as well as on SHP-1/STAT3 signaling in RMCs and HEK293 cells. We also investigated the role of SHP-1 overexpression in IFN-treated RMCs and HEK293 cells; SHP-1 overexpression resulted in a similar phenotype as that observed with circ_0007059 expression. Conclusions The study indicates that circ_0007059 protects RMCs against apoptosis and inflammation during nephritis by attenuating miR-1278/SHP-1/STAT3 signaling.

Abstract Background Ferroptosis is a mode of regulated cell death that depends on iron, plays pivotal roles in regulating various biological process in human cancers. However, the role of ferroptosis in Gastric cancer (GC) remains unclear. Methods A total of 2721 differentially expressed genes (DEGs) were filtered base on The Cancer Genome Atlas (TCGA) (n = 375) dataset. Gene modules were identified based on co-expression network analysis (WGCNA). Functional analysis was performed to explore the biological function. Lasso-penalized and univariate Cox regression (UCR) analysis, survival genes were screened out to construct a prognostic model, which validated by the GSE43292 dataset. Gene set enrichment analysis (GSEA) for prognostic index was performed. Finally, the correlations of ferroptosis and immune cells were assessed through the TIMER database. Results Compared to normal specimens, 1063 highly upregulated and 1658 downregulated genes respectively and their normal counterparts in GC specimens were screened. WGCNA analysis was used and identified 7 modules, of which, blue module with the most significant enrichment result was selected. By taking intersections of blue module and differentially expressed ferroptosis-related genes (DEFRGs), we got 23 common genes. Functional analysis was performed to explore the biological function of the interested genes, and with the consequences Lasso-penalized and univariate Cox regression (UCR) analysis, survival genes were screened out to construct a prognostic model based on 3 genes (SLC1A5, ANGPTL4, and CGAS), which could play a role in predicting the survival of GC patients. UCR and multivariate Cox regression (MCR) analysis revealed that the prognostic index could be used as independent prognostic indicators and validated using another GSE84437 dataset. Notably, patients in high-risk groups had higher levels of higher mutation frequencies such as TTN and TP53.Mechanistically. Gene set enrichment analysis (GSEA) unveiled several significant and pathways involved in GC. TIMER analysis demonstrated that risk score strongly correlated with Macrophage and CD4 + T cells infiltration. In addition, high- and low-risk group illustrated different distributions in different immune status. Conclusions In this study, a novel FRGs signature was built. It could accurately predict GC prognosis and pave the new way for diagnosis and therapy strategy. May reflect the status of tumor immune microenvironment.

Diabetic kidney disease (DKD) is a severe form of microangiopathy among diabetic patients, of which podocyte injury is one of the more predominant features. There is increasing evidence to suggest that mitochondrial dysfunction is associated with podocyte injury, thus contributing to the progression of DKD. Initially identified as a p53 target protein, the endogenous antioxidant protein, sestrin-2 (sesn2), has recently attracted attention due to its potential function in various inflammatory diseases. However, the association between sesn2 and podocytes in DKD remains unclear. In the present study, to elucidate the role of sesn2 in podocyte mitochondrial dysfunction, the effects of sesn2 on the regulation of AMP-activated protein kinase (AMPK) were examined in vitro and in vivo. Abnormal mitochondria were found in rats with streptozotocin-induced diabetes, and hyperglycemia downregulated the expression of sesn2. The upregulation of sesn2 increased the level of AMPK phosphorylation, and thus ameliorated mitochondrial dysfunction under high glucose conditions (HG). On the whole, these results suggest that sesn2 is associated with mitochondrial dysfunction in podocytes under HG conditions. In addition, the decreased expression of sesn2 may be a therapeutic target for DKD.

Reactive oxygen species (ROS) impair neovascularization and perfusion recovery following limb ischemia in patients with peripheral arterial disease (PAD). Hydrogen molecules (H2) comprise an antioxidant gas that has been reported to neutralize cytotoxic ROS. The present study investigated whether H2 may serve as a novel therapeutic strategy for PAD. H2-saturated water or dehydrogenized water was supplied to mice with experimental PAD. Laser Doppler perfusion imaging demonstrated that H2-saturated water improved perfusion recovery, decreased the rate of necrosis, increased the capillary density in the gastrocnemius muscle and increased the artery density in the abductor muscle in the ischemic limbs, at 14 and 21 days post-hindlimb ischemia. Ischemic muscle tissue was harvested 7 days after experimental PAD for biochemical testing and H2 was observed to reduce the levels of malondialdehyde and increase the levels of cyclic guanine monophosphate (cGMP). In cultured endothelial cells, H2-saturated culture medium resulted in reduced ROS levels, increased tube formation and increased cGMP levels. In macrophages, H2 decreased cellular ROS levels and promoted M2 polarization. H2-saturated water increases angiogenesis and arteriogenesis and subsequently improves perfusion recovery in a mouse PAD model via reduction of ROS levels.

Esophageal squamous cell carcinoma (SCC) possesses one of the worst prognoses out of the digestive carcinomas. Several studies have suggested that transforming growth factor β receptor type II (TGF-βRII), Smad family member 4 (Smad4) and p21 wild-type p53-activated factor 1 (p21waf1) are associated with esophageal SCC. The aim of the present study was to evaluate the effect of Smad4, TGF-βRII and p21waf1 in esophageal squamous cancer tissue and the pathological significance of the effect. An immunohistochemical method was used to evaluate the expression levels of Smad4, TGF-βRII and p21waf1 in specimens of esophageal SCC lesions obtained from 80 patients. It was found that the expression of Smad4, TGF-βRII and p21waf1 in histologically-classified grade I esophageal SCC, without invasion or lymph node metastasis, was markedly higher compared with grade III esophageal SCC that had invaded into the deep muscular or serous layer and metastasized to the lymph nodes (P<0.05). Analysis of the expression level of Smad4, TGF-βRII and p21waf1, as well as the clinical and pathological characteristics of esophageal SCC, revealed that the three proteins may be associated with the carcinogenesis, biological behavior and prognosis of esophageal SCC, parallel to the pathological stage and cell grade.

PCBs are a family of persistent environmental toxicants with a wide spectrum of toxic features, such as neurotoxic, hepatoxicity, immunotoxicity, endocrine disruption effects, and oncogenic effects. The kidney is the most important organ involved in the elimination of toxins and drugs. To date, little has been done to investigate the potential influence of nephrotoxicity of 3,3’,4,4’- tetrachlorobiphenyl (PCB77). By assessing cell viability and apoptotic cell death in renal tubular epithelial (NRK-52E) cells cultures, we found that PCB77 could decrease cellular viability at least at 30 μM concentration after 3 h exposure. PCB77 was demonstrated to promote DNA breakage resulting in apoptosis. Moreover, apoptotic subcellular morphological changes administration of PCB77 was observed using transmission electron microscopy. Appearance swelling of mitochondria, endoplasmic reticulum dilation and chromatin agglutinate, and other apoptosis cells morphological characteristics could be visible. Due to increased PCB77 concentration, cells viability was decreased. Collectively, our findings identified the morphological mechanism that PCB77-induced nephrotoxicity via promoting renal tubular epithelial cells apoptosis. It is suggested that using and production of PCB77 should be carefully managed to reduce public health risks.

OBJECTIVE:To investigate the effect of urokinase-type plasminogen activator (uPA) on mesangial matrix in the kidney of diabetic rats and its related mechanisms. METHODS:Diabetic Sprague-Dawley (SD) rats induced by intraperitoneal injection of streptozotocin (STZ) were randomly and evenly divided into two groups: DM + vehicle, and DM + uPA (2500 U kg(-1) uPA via tail vein once a day for four weeks). The normal SD rats without diabetes were considered as control group. Rats in the three groups were executed and the heart blood was sampled for determination of blood glucose and serum creatinine. Meanwhile, kidney tissues of rats were also harvest for measurement of glomerular area, volume, and mesangial area by periodic acid silver methenamine (PASA) staining. The expression of urokinase-type plasminogen activator receptor (uPAR), plasminogen activator inhibitor-1 (PAI-1), and collagen IV in renal tissues was tested with immunohistochemistry. RESULTS:Compared with control, the DM rats had obvious albuminuria, significantly (p < 0.01) increased glomerular volume and mesangial matrix area, and significantly (p < 0.05) higher expression of uPAR, PAI-1 and collagen IV in mesangial matrix, significantly up-regulated (p < 0.05) glomerular uPAR, PAI-1, and collagen IV expression. After treated with uPA, the diabetic rats had significantly (p < 0.05) reduced albuminuria, significantly (p < 0.01) improved glomerular volume and mesangial matrix, significantly (p < 0.05) down-regulated PAI-1 and collagen IV expression in mesangial matrix. However, the uPAR expression in renal tissues were unchangeable (p > 0.05) and PAI-1 and collagen IV expression were significantly (p < 0.05) reduced when diabetic rats were treated with uPA. CONCLUSION:uPA can down-regulate glomerular PAI-1 expression in the DM rats but not significantly influence uPAR expression, suggesting that uPA might regulate the mesangial cell (MC) and its matrix expression and improve diseased diabetic mesangial matrix via its combination with uPAR to uptake PAI-1 and accelerate its degradation.

Objective: To evaluate the effect of urokinase-type plasminogen activator (uPA) on high-glucose induced-proliferation and alteration of α-SMA expression of glomerular mesangial cell (MC). Methods: MCs were exposed to high-glucose (30 mmol/L D-glucose) medium or high glucose plus different concentration of uPA. The cell cycle and proliferation were respectively assessed by flow cytometry and MTT assay. Laser scanning confocal microscope was used to observe the expression of α-SMA. Results: uPA prevented high glucose-exposed MC from G0/G1 phase into S phase in a dose-dependent manner, thus inhibited proliferation of MC. High glucose exposure increased perinuclear expression of α-SMA in MC, which was prevented by uPA in a dose-dependant manner. Conclusion: uPA may exert a therapeutic effect on diabetic glomerulosclerosis through inhibiting the proliferation and phenotypic change of MC.

The Rho kinase pathway plays an important role in epithelial dedifferentiation and inflammatory cell infiltration. Recent studies suggest that inflammation promotes lymphangiogenesis, which has been associated with renal allograft rejection. We investigated whether targeted inhibition of the Rho kinase pathway in proximal tubular cells reduces inflammation and lymphangiogenesis in acute renal allograft rejection. The Rho kinase inhibitor Y27632 was coupled to lysozyme (Y27632-lysozyme), providing a kidney-specific conjugate that can release its drug in proximal tubular cells. Isogenic (Fisher–Fisher, n=18), or allogenic (Fisher–Lewis, n=24) kidney transplantations were performed, with the contralateral kidney remaining in situ. To elicit acute rejection, no immunosuppressive treatment was given. Animals were treated daily with Y27632-lysozyme (10mg/kg/day i.v.) or vehicle (saline i.v.) until sacrifice (1 or 4 days post-transplantation). After allogenic transplantation, interstitial macrophage accumulation was strongly reduced by Y27632-lysozyme at day 4 after transplantation. Interstitial lymphangiogenesis, which was induced in allografts as compared to control kidney, was also reduced by renal Rho kinase inhibition at day 4 after transplantation. The increase of vimentin and procollagen-1alpha1 gene expression in renal allografts from day 1 to day 4 after transplantation was significantly reduced by Y27632-lysozyme. Y27632-lysozyme did not affect systolic blood pressure in isogenic or allogenic transplantation groups. In cultured tubular epithelial cells (NRK-52E), Rho kinase inhibition dose-dependently reduced IL-1β-induced MCP-1 gene expression. Renal inhibition of Rho kinase causes a marked reduction in renal inflammation and renal lymphangiogenesis during acute transplant rejection, suggesting that this treatment regimen is a valuable future treatment in renal transplantation.

Objective: To evaluated the association between the development of proteinuria and angiotensin Ⅱ infusion and investigate the morphological changes in podocyte foot process and the expression of the glomerular podocyte molecule,nephrin.Methods: Wistar rats were randomly assigned into twogroups according to receiving either normal saline(control) or AngⅡ at a dose of 400 ng/(kg·min) via subcutaneous osmotic mini-pumps.Proteinuria was measured before rats were sacrificed at each week.Renal morphological changes were evaluated by light and transmission(electron) microscopy.Semiquantitative reverse transcription-polymerase chain reaction(RT-PCR) and immunofluorescence(IF) staining were performed to study the glomerular expression of nephrin.Results: Rats with Ang Ⅱ infusion developed proteinuria markedly were compared with control rats.The capsules of glomeruli in Ang Ⅱ group became thinner at the 7th day and electron microscopy revealed flourish podocyte foot processes.The intensity of nephrin expression increased significantly and the negative expression was localized at glomerular epithelial cells in AngⅡ group after 14 days(by 58.9% more than in control,P0.01) and no linear staining could be observed.During the increasing of proteinuria,the foot process became wider at 21 d and nephrin decreased significantly both in protein intensity and at mRNA level.Conclusion: During the onset of AngⅡ,the first response of podocyte is the flourish foot process as well as the distribution change of nephrin.Ang Ⅱ increases the expression of nephrin.There was a close relationship between nephrin distribution pattern with the development of proteinuria and with the podocyte foot process changes.Proteinuria might occur after series of events of nephrin distribution changes and morphologic changes of podocyte foot process.