BACKGROUND:Calcium oxalate (CaOx) kidney stones are the most common form of urolithiasis and are characterized by high incidence and recurrence rates. 4-Octyl itaconate (4-OI), an itaconate derivative, has been shown to exert broad protective effects in various disease models owing to its anti-inflammatory and antioxidant properties. However, its role in CaOx kidney stone formation remains largely unknown. METHODS AND RESULTS:A CaOx kidney stone model was established in 6-week-old male C57BL/6 mice by glyoxylic acid induction, followed by treatment with 4-OI at 50 mg/kg. Renal tissues were subjected to HE, TUNEL, DHE, Prussian blue, and von Kossa staining to evaluate renal tissue injury, apoptosis, reactive oxygen species (ROS) production, iron deposition, and CaOx crystal deposition. Mechanistically, we observed that 4-OI exerts its effects through the NRF2-HO-1/SLC7A11 pathway in HK-2 cells. Our results suggest that 4-OI prevents CaOx crystal formation, attenuates oxidative stress, and mitigates ferroptosis both in vitro and in vivo via the NRF2-HO-1/SLC7A11 pathway. CONCLUSION:4-OI attenuates renal CaOx formation, reduces oxidative stress, and alleviates ferroptosis through the NRF2-HO-1/SLC7A11 pathway. These findings suggest that 4-OI has significant therapeutic potential for the treatment of CaOx kidney stones.

BACKGROUND:Cancer stem cells (CSCs) are key contributors to treatment failure in clear cell renal cell carcinoma (ccRCC). Our prior single-cell RNA-seq analyses identified CSCs as the predominant source of SPP1, which may maintain stemness and promote progression via JAK1/STAT3 signaling, with POU5F1 as a putative upstream regulator. However, these SPP1-centered roles and their upstream/downstream mechanisms remain to be systematically validated. METHODS:We integrated bioinformatics analysis with experimental validation using functional assays (proliferation, migration, tumorsphere formation), molecular mechanism studies (ChIP, Co-IP, dual-luciferase assays), and in vivo xenograft models to investigate the roles and mechanisms of SPP1. RESULTS:In 30 paired tissues, POU5F1 and SPP1 were upregulated in ccRCC, and concurrent high expression in TCGA predicted poorer overall survival. In 786-O/769-P cells, loss- and gain-of-function experiments showed that either gene modulated proliferation, EdU incorporation, migration, and tumorsphere formation. SPP1 silencing reduced pJAK1/pSTAT3, whereas STAT3 blockade partially reversed SPP1-induced phenotypes. POU5F1 occupied the SPP1 promoter and increased promoter-driven transcription, leading to higher SPP1 expression and secretion; SPP1 knockdown attenuated POU5F1-overexpression effects. In xenografts, lowering POU5F1 or SPP1 decreased tumor growth, Ki-67, stem-like/EMT markers, and JAK1/STAT3 activity. CONCLUSIONS:Our findings support a POU5F1-SPP1- JAK1/STAT3 axis that sustains stem-like traits and malignant behavior in ccRCC. Targeting this axis may concurrently suppress bulk tumor cells and CSC compartments, providing a biomarker-guided, dual-hit therapeutic rationale.

Background:Acute kidney injury (AKI) is a prevalent complication of sepsis, where the inflammatory response plays a crucial role. Selenium exhibits anti-inflammatory and antioxidant properties, but its impact on sepsis-induced AKI remains unclear. Methods and results:In this study, we used a lipopolysaccharide (LPS)-induced murine model of sepsis-associated acute kidney injury (SA-AKI) in male C57BL/6 mice (8-12 weeks old) to investigate the protective mechanisms of selenomethionine (SeMet). Mice received weekly oral administration of SeMet (0.375 mg/kg) commencing 1 week prior to AKI induction. Our results demonstrated that SeMet treatment significantly attenuated the inflammatory response, reduced oxidative stress, and ameliorated renal pathological damage compared to saline-treated controls. Mechanistic investigations revealed that SeMet modulates altered mitochondrial dynamics and suppresses the NF-κB signaling pathway, thereby promoting macrophage polarization toward the anti-inflammatory M2 phenotype. Conclusion:These findings collectively demonstrate that SeMet effectively mitigates inflammation and ameliorates sepsis-induced AKI, suggesting its potential as a therapeutic candidate for SA-AKI prevention and treatment.

Objectives: To investigate the role of selenium and selenium-containing proteins in the etiology and pathogenesis of kidney stones.Methods: The HK-2 cell line was subjected to supersaturation oxalate treatment to establish an in vitro model of calcium oxalate kidney stones, while SD rats were administered with ethylene glycol to establish an in vivo model of calcium oxalate kidney stones. qPCR analysis was employed to investigate the alterations in selenoproteins within the models, and subsequently, genes exhibiting significant changes were identified. Subsequently, based on the functions of these genes, their regulatory effects on endoplasmic reticulum stress (ERS) and apoptosis during the disease progression were examined both in HK-2 cells and rat kidneys. Finally, Selenomethionine (SeMet) supplementation was introduced to explore its therapeutic potential for kidney stone management.Results: The involvement of Selenoprotein K in the pathogenesis of calcium oxalate kidney stone disease has been confirmed, exhibiting significant alterations. Manipulation of its expression levels through overexpression and knockdown techniques resulted in a corresponding reduction or increase in oxidative stress, ERS, and apoptosis within renal tubular epithelial cells. SelK regulates ERS and apoptosis by controlling the IRE1-ASK1-JNK pathway. In addition, SeMet treatment, which contains selenium, effectively reduced the levels of oxidative stress, ERS, and apoptosis in vivo and in vitro models, thereby alleviating tubular epithelial cell damage and reducing the formation of kidney stones in experimental rats.Discussion: Selenium is involved in the occurrence and development of kidney stones by regulating oxidative damage to renal tubular epithelial cells. The results suggest that dietary selenium supplementation in daily life may be of great significance for the prevention and treatment of kidney stones.

Immune checkpoint inhibitors (ICIs) are a cornerstone therapy for advanced renal cell carcinoma (RCC). However, significant rates of primary resistance hinder their efficacy, and the underlying mechanisms remain poorly understood. This study aims to unravel the tumor-immune interactions and signaling pathways driving primary resistance to ICIs in RCC. We integrated single-cell RNA sequencing, spatial transcriptomics, and clinical sample analysis to investigate the tumor microenvironment and intercellular signaling. Advanced computational methods, including cell–cell communication networks, pseudotime trajectories, and gene set enrichment analysis (GSEA), were employed to uncover the underlying resistance mechanisms. Compared to the sensitive group, the primary resistance group exhibited a significant increase in SPP1-CD44 signaling-mediated interactions between tumor cells and immune cells. These interactions disrupted antigen presentation in immune effector cells and suppressed key chemokine and cytokine pathways, thereby impairing effective immune responses. In contrast, the sensitive group showed more active antigen presentation and cytokine signaling, which facilitated stronger immune responses. Furthermore, the interaction between SPP1-secreting tumor cells and CD44-expressing exhausted CD8 + T cells activated the MAPK signaling pathway within CD8 + Tex cells, exacerbating T cell exhaustion and driving the development of ICI resistance in RCC. Our findings reveal a potential mechanism by which SPP1-CD44 signaling mediates tumor-immune cell interactions leading to ICI resistance, providing a theoretical basis for targeting and disrupting this signaling to overcome primary resistance in RCC.

To analyze the relationship between the composition of urinary stones and various influencing factors in the Enshi region. We used FT-IR to examine the composition of 1092 stone samples. Combined with the relevant clinical materials, the data were analyzed using both one-dimensional statistical methods and multivariate statistical methods. The study included 1092 stone samples, classified as follows: 457 (41.8%) with a single component, 453 (41.5%) with two components, 149 (13.6%) with three components, and 33 (3.0%) with four components. Stones were categorized into five types: Calcium Oxalate (CaOx) (76.4%), carbapatite (CaP) (9.3%), Struvite (ST) (8.3%), Uric Acid (UA) (4.9%), and Others (1.0%). Age, gender, urinary tract infection (UTI), family history of urinary stones (FH), hyperuricemia (HUA) and stone location were significantly associated with stone type. Logistic regression revealed that females and UTI were relative risk factors for predicting CaP and ST, while FH and HUA were relative risk factors for predicting UA. Our study indicates that the overall composition of urinary tract stones in the Enshi region is consistent with that of the entire China. Additionally, the predisposing factors for stone formation vary in terms of gender, age, FH, UTI, hyperuricemia HUA, and stone location.

Objective: This study aims to explore the mechanism of puerarin regulating the proliferation, apoptosis and senescence of T24 and EJ cells via miR-139-5p/CCNB1 and PI3K/AKT pathways. Methods: Initially, Cell Counting Kit (CCK)-8, flow cytometry, beta-galactosidase detection kit, and RT-qPCR are used to assess the effect of puerarin on the proliferation apoptosis and senescence of bladder cancer cells by regulating miR-139-5p, respectively. Then, potential complementary binding sites between miR-139-5p and the 3 ' untranslated region (3 ' UTR) of the CCNB1 gene are obtained by the bioinformatics software, starBase database. Further, a dual-luciferase reporter system is used to assess the targeted relationship of miR-139-5p to CCNB1. CCK-8, flow cytometry, and beta-galactosidase detection kit were used to detect the influence of CCNB1 on the proliferation, apoptosis, and senescence of bladder cancer cells. RT-qPCR and Western blotting are used to determine the regulation of CCNB1 and PI3K/AKT pathways by miR-139-5p. Finally, the subcutaneous tumor formation in nude mice and immunohistochemical experiments are performed to assess the effect of puerarin on the proliferation of bladder cancer cells in vivo. Results: Puerarin induced proliferation inhibition, apoptosis, and senescence of bladder cancer cells in vitro. Puerarin down-regulated the expression of CCNB1, p-PI3K, and p-AKT proteins by overexpressing miR-139-5p, thereby exerting a tumor suppressor effect in bladder cancer. Compared with the control group, the volume and weight of subcutaneous tumors of nude mice in the puerarin administration group are reduced. Moreover, the ki67 positive staining rate and the expression level of CCNB1, p-PI3K, and p-AKT protein are reduced. Conclusion: The experimental results showed that the anti-tumor effect of puerarin against bladder cancer might be achieved by inhibiting CCNB1 and PI3K/AKT pathway via up-regulating miR-139-5p.

Background: Muscle invasive bladder cancer (MIBC) is one of the most common malignant diseases in elderly men, such as veterans. Postoperative chemotherapy plays a vital role in preventing recurrence and metastasis of MIBC. However, fewer of chemotherapeutic drugs with remarkable anti-tumor activity and biosafety are available for clinical treatment. Methods: In this work, a novel plant-derived drug, named as dihydromyricetin (DHM), was selected for postoperative chemotherapy of MIBC. The anti-tumor activity was evaluated using a series of in vitro tests, such as MTT assay, flow cytometry and western blot. Furthermore, a xenograft model of BALB/C57 nude mice was established to verify the good anti-tumor activity and biosafety of DHM in vivo. Results: DHM could effectively inhibit the proliferation, survival and migration of MIBC cell, and promote apoptosis (P<0.05). Cytotoxic macrophage polarization from M0 to M1 was promoted by DHM treatment, and the hub genes in cell cycle and apoptosis signaling pathways were differential expressed. We also found that DHM could reverse the epithelial-mesenchymal transition (EMT) of MIBC cell. The in vivo results revealed that intravenous injection of DHM with a dose of 20 mg/kg for 7 times could significantly suppress the in vivo tumorigenesis of MIBC (P<0.05), while triggered no obvious drug side effects. Conclusion: This work identified a novel chemotherapeutic DHM with remarkable anti-tumor activity and biosafety, which could serve as a promising alternative for postoperative chemotherapy of MIBC.

Zijian Wang Wuhan University Zhongnan Hospital Zicheng Guo Wuhan University Zhongnan Hospital Wenjie You Wuhan University Zhongnan Hospital Wang Wang Wuhan University Zhongnan Hospital Fenfang Zhou Wuhan University Zhongnan Hospital Renjie Zhang Wuhan University Zhongnan Hospital Zhiwen He Wuhan University Zhongnan Hospital Hongbo Chen enshi tujiazu miaozu zizhizhou zhongxin yiyuan: Central Hospital of Enshi Tujia and Miao Autonomous Prefecture Xinghuan Wang (  wangxinghuan@whu.edu.cn ) Wuhan University Zhongnan Hospital https://orcid.org/0000-0003-3497-0024

Objective To evaluate the effect and mechanism of dihydromyricetin (DHM) on the proliferation of human bladder cancer cells.Methods UMUC3 cells were incubated with DHM at the concentrations of 0,5 and 20 μmol/L respectively,then the inhibitory effect were investigated by thiazole blue (MTT) assay and flow cytometry.The expression of Cyclin D1 and Cyclin E1 and cyclin-dependent kinases (CDK2 and CDK4) was detected using real-time fluorescence quantitativepolymerase chain reaction (FQ-PCR) and Western blotting.Statistical analysis was performed using SPSS 21.0 software and the results were expressed by mean value ± standard deviation (Mean ± SD).The statistical method was one-way analysis of variance incorporated with the least significant difference (LSD) method and Duncan multiple range test.Results DHM inhibited the proliferation of UMUC3 cells obviously.Compared with the control group [(48.4 ± 0.6)％],the percentage of G0/G1 phase cells in low concentration group [(55.2 ± 0.1) ％] and high concentration group [(60.5 ± 0.3) ％] increased,and the difference was statistically significant (F=417.668 in the low concentration group,P ＜0.01,F=401.652 in the high concentration group,P ＜ 0.01).The percentage of G0/G1 phase cells was higher in the high concentration group than in the low concentration group,and the difference was statistically significant (F =100.221,P ＜ 0.05).The results of RT-PCR and Western blotting showed that the expression of CDK2,CDK4,Cyclin D1 and Cyclin E1 decreased obviously.Conclusion DHM can effectively inhibit the proliferation of human bladder cancer cells,and its mechanism may be related to blocking the transition of cell cycle from G0/G1 phase to S phase.