The SOX4/EZH2/SLC7A11 signaling axis mediates ferroptosis in calcium oxalate crystal deposition-induced kidney injury.

Epigenetic regulation is reported to play a significant role in the pathogenesis of various kidney diseases, including renal cell carcinoma, acute kidney injury, renal fibrosis, diabetic nephropathy, and lupus nephritis. However, the role of epigenetic regulation in calcium oxalate (CaOx) crystal deposition-induced kidney injury remains unclear. Our study demonstrated that the upregulation of enhancer of zeste homolog 2 (EZH2)-mediated ferroptosis facilitates CaOx-induced kidney injury. CaOx crystal deposition promoted ferroptosis in vivo and in vitro. Usage of liproxstatin-1 (Lip-1), a ferroptosis inhibitor, mitigated CaOx-induced kidney damage. Single-nucleus RNA-sequencing, RNA-sequencing, immunohistochemical and western blotting analyses revealed that EZH2 was upregulated in kidney stone patients, kidney stone mice, and oxalate-stimulated HK-2 cells. Experiments involving in vivo EZH2 knockout, in vitro EZH2 knockdown, and in vivo GSK-126 (an EZH2 inhibitor) treatment confirmed the protective effects of EZH2 inhibition on kidney injury and ferroptosis. Mechanistically, the results of RNA-sequencing and chromatin immunoprecipitation assays demonstrated that EZH2 regulates ferroptosis by suppressing solute carrier family 7, member 11 (SLC7A11) expression through trimethylation of histone H3 lysine 27 (H3K27me3) modification. Additionally, SOX4 regulated ferroptosis by directly modulating EZH2 expression. Thus, this study demonstrated that SOX4 facilitates ferroptosis in CaOx-induced kidney injury through EZH2/H3K27me3-mediated suppression of SLC7A11.

Gut microbiota in patients with kidney stones: a systematic review and meta-analysis.

BACKGROUND: Mounting evidence indicates that the gut microbiome (GMB) plays an essential role in kidney stone (KS) formation. In this study, we conducted a systematic review and meta-analysis to compare the composition of gut microbiota in kidney stone patients and healthy individuals, and further understand the role of gut microbiota in nephrolithiasis. RESULTS: Six databases were searched to find taxonomy-based comparison studies on the GMB until September 2022. Meta-analyses were performed using RevMan 5.3 to estimate the overall relative abundance of gut microbiota in KS patients and healthy subjects. Eight studies were included with 356 nephrolithiasis patients and 347 healthy subjects. The meta-analysis suggested that KS patients had a higher abundance of Bacteroides (35.11% vs 21.25%, Z = 3.56, P = 0.0004) and Escherichia_Shigella (4.39% vs 1.78%, Z = 3.23, P = 0.001), and a lower abundance of Prevotella_9 (8.41% vs 10.65%, Z = 4.49, P < 0.00001). Qualitative analysis revealed that beta-diversity was different between the two groups (P < 0.05); Ten taxa (Bacteroides, Phascolarctobacterium, Faecalibacterium, Flavobacterium, Akkermansia, Lactobacillus, Escherichia coli, Rhodobacter and Gordonia) helped the detection of kidney stones (P < 0.05); Genes or protein families of the GMB involved in oxalate degradation, glycan synthesis, and energy metabolism were altered in patients (P < 0.05). CONCLUSIONS: There is a characteristic gut microbiota dysbiosis in kidney stone patients. Individualized therapies like microbial supplementation, probiotic or synbiotic preparations and adjusted diet patterns based on individual gut microbial characteristics of patients may be more effective in preventing stone formation and recurrence.

STAT6 promoting oxalate crystal deposition-induced renal fibrosis by mediating macrophage-to-myofibroblast transition via inhibiting fatty acid oxidation.

OBJECTIVE AND DESIGN: Kidney stones commonly occur with a 50% recurrence rate within 5 years, and can elevate the risk of chronic kidney disease. Macrophage-to-myofibroblast transition (MMT) is a newly discovered mechanism that leads to progressive fibrosis in different forms of kidney disease. In this study, we aimed to investigate the role of MMT in renal fibrosis in glyoxylate-induced kidney stone mice and the mechanism by which signal transducer and activator of transcription 6 (STAT6) regulates MMT. METHODS: We collected non-functioning kidneys from patients with stones, established glyoxylate-induced calcium oxalate stone mice model and treated AS1517499 every other day in the treatment group, and constructed a STAT6-knockout RAW264.7 cell line. We first screened the enrichment pathway of the model by transcriptome sequencing; detected renal injury and fibrosis by hematoxylin eosin staining, Von Kossa staining and Sirius red staining; detected MMT levels by multiplexed immunofluorescence and flow cytometry; and verified the binding site of STAT6 at the PPARα promoter by chromatin immunoprecipitation. Fatty acid oxidation (FAO) and fibrosis-related genes were detected by western blot and real-time quantitative polymerase chain reaction. RESULTS: In this study, we found that FAO was downregulated, macrophages converted to myofibroblasts, and STAT6 expression was elevated in stone patients and glyoxylate-induced kidney stone mice. The promotion of FAO in macrophages attenuated MMT and upregulated fibrosis-related genes induced by calcium oxalate treatment. Further, inhibition of peroxisome proliferator-activated receptor-α (PPARα) eliminated the effect of STAT6 deletion on FAO and fibrosis-associated protein expression. Pharmacological inhibition of STAT6 also prevented the development of renal injury, lipid accumulation, MMT, and renal fibrosis. Mechanistically, STAT6 transcriptionally represses PPARα and FAO through cis-inducible elements located in the promoter region of the gene, thereby promoting MMT and renal fibrosis. CONCLUSIONS: These findings establish a role for STAT6 in kidney stone injury-induced renal fibrosis, and suggest that STAT6 may be a therapeutic target for progressive renal fibrosis in patients with nephrolithiasis.

Identification of amino acid residues that are crucial for FXIII-A intersubunit interactions and stability.

Coagulation factor XIII (FXIII) is the main stabilizer of the fibrin clot. It circulates in plasma as a tetramer of two A-subunits and two B-subunits. Under physiological conditions, FXIII-A exists as a dimer (FXIII-A2). The interactions between the FXIII-A-subunits that stabilize the FXIII-A2 dimer are not fully understood. We therefore designed a systematic approach to identify amino acid residues crucial for the expression and stability of FXIII-A2. Based on the available FXIII-A2 crystal structure, we identified 12 amino acid residues forming intersubunit salt bridges and 21 amino acid residues forming hydrogen bonds between the two A-subunits. We chose 10 amino acid residues that form 5 particularly strong interactions, performed site-directed mutagenesis, and expressed the mutants in CHO cells. Disruption of these interactions by single mutation of Lys257, Lys113, Asp343, Glu401, or Asp404 abolished the expression of properly folded, soluble, and functional FXIII-A in CHO cells. On the contrary, mutation of Glu111, Arg100, or Asn112 had no significant effect on FXIII-A expression. Our results suggest that 4 intersubunit interactions (Arg11-Asp343, Lys113-Asp367, Lys257-Glu401, and Arg260-Asp404) are essential for the stability of FXIII-A2. Our findings are supported by reported mutations at Lys257, Arg260, and Asp404 found in patients with congenital FXIII-A deficiency.

Preparation of hollow-fiber nanofiltration membranes of high performance for effective removal of PFOA and high resistance to BSA fouling.

Nanofiltration (NF) process has become one of the most promising technologies to remove micro-organic combined water pollution. Developing a NF membrane material with efficient separation for perfluorooctanoic acid (PFOA) combined pollution is highly desired, this manuscript targets this unmet need specifically. In this work, hydrophilic SiO2 nanoparticles with various contents blended with carboxylic multiwalled carbon nanotube were used to modify poly (m-phenylene isophthal amide) (SiO2/CMWCNT/PMIA) hollow fiber NF membrane. The modified membrane with 0.1 wt% SiO2 doping exhibits way better fouling resistance with irreversible fouling ratio decreased dramatically from 18.7% to 2.3%, and the recovery rate of water flux increases significantly from 81.2% to 97.7%. The separation experiment results had confirmed that the modified membrane could improve the rejection from 97.2% to 98.6% for perfluorooctanoic acid (PFOA) and its combined pollution with bovine serum albumin (BSA). It is clear that this reported SiO2/CMWCNT/PMIA hollow fiber NF membrane potentially could be applied in water treatment. This research also provides a theoretical basis for efficiently removal of PFOA and its combined pollution by NF membrane.

One-Step Electrochemically Prepared Graphene/Polyaniline Conductive Filter Membrane for Permeation Enhancement by Fouling Mitigation.

In the electrofiltration process, membrane conductivity plays a decisive role in improving the antifouling performance of the membrane. In this paper, combining the preparation of graphene (Gr) with the fabrication of the Gr layer on the surface of a polyaniline (PANI) membrane, a graphene/PANI (Gr/PANI) conductive membrane was prepared creatively by the one-step electrochemical method. The properties of the as-prepared Gr/PANI membrane were studied systematically. By the tests of Fourier transform infrared spectroscopy, Raman spectroscopy, X-ray diffraction, and atomic force microscopy, it was confirmed that Gr was successfully produced and was combined with the PANI membrane well. Field scanning electron microscopy with energy-dispersive X-ray analysis further confirmed that the top surface and the upper layer pore walls of the membrane were randomly covered by Gr. The antifouling performance of the prepared membrane was evaluated by studying the permeation flux of the yeast suspension, compared with the ones with no electric field: the total permeation flux at 1 V direct current (dc) increased by 109%; besides, under 1 V dc, the average flux of the Gr/PANI membrane was approximately 1.4 times that of the PANI membrane. This approach may provide a promising strategy for the combination of Gr with conductive polymers to produce separation membranes.

Left atrial appendage exclusion is effective in reducing postoperative stroke after mitral valve replacement.

OBJECTIVE: This study aimed to evaluate the role of surgical left atrial appendage (LAA) exclusion in the prevention of stroke after mitral valve replacement (MVR). METHODS: We retrospectively reviewed clinical data of 860 patients who received MVR in our center from January 2008 to January 2013. The patients were randomly assigned to two surgical groups, namely LAA exclusion group (n = 521) and LAA nonexclusion group (n = 339) according to whether concurrent surgical exclusion of the LAA was to be undertaken or not before surgery in a blind fashion. MVR was performed by two experienced surgeons. The LAA was explored during the operation and mural thrombus removed in all cases. The LAA was left intact in nonocclusion group whereas the neck of the LAA was closed with a two-layer continued suture in exclusion group. The incidence of early postoperative ischemic stroke between the two groups was compared. RESULTS: The patients' age was 53 ± 12 years, with 48.1% male and 67.9% with rheumatic disease. Mural thrombosis was seen in 18.8% of the patients and atrial fibrillation (AF) coexisted in 62.4%. All operations were successfully performed and no difference was noted in in-hospital mortality, re-exploration for bleeding, and other major complications between the two groups. The incidence of ischemic stroke in LAA exclusion group was significantly lower than in nonexclusion group (0.6% vs. 2.7%, p = .011). The subgroup multivariate analysis showed that LAA exclusion significantly reduced the risk of postoperative stroke in patients with AF (odds ratio [OR] = 0.070, 95% confidence interval [CI]: 0.006-0.705, p = .025) but not in non-AF patients (OR = 1.902, 95% CI: 0.171-21.191, p = .601). CONCLUSIONS: Concurrent LAA exclusion during MVR is a safe and effective way to reduce postoperative ischemic stroke, particularly in patients with AF.

[Early results of left atrial appendage closure in cerebral ischemic stroke reduction in patients with mitral valve replacement].

OBJECTIVE: To investigate the role of left atrial appendage (LAA) closure for cerebral ischemic stroke prevention following mitral valve replacement. METHODS: Retrospective data on 860 consecutive adult patients undergoing mitral valve replacement between January 2008 and January 2013 were analyzed. There were 414 male and 446 female patients, with a mean age of (53 ± 12) years. The patients were divided into two groups according to whether the left atrial appendage was closed during operation: LAA closure group (n = 521) and non-LAA closure group (n = 339).Early mortality, postoperative cerebral ischemic stroke and the risk factors for cerebral ischemic stroke were assessed. Multivariate analysis was performed using logistic regression analysis. RESULTS: Compared with non-LAA closure group, LAA closure group had higher proportion of female gender, higher percentage of patients with cardiac insufficiency, pulmonary hypertension and left atrial thrombus, higher incidence of mechanical valve implantation and concurrent tricuspid surgery, and larger preoperative diameter of left atrium, but lower proportion of hypertension and patients undergoing coronary artery bypass surgery, and shorter aorta cross clamping time (χ² = 6.807 to 122.576, t = -2.818 and 3.756, all P < 0.05). There were no differences in exploratory thoracotomy for bleeding and in-hospital mortality between the two groups. Postoperative cerebral ischemic stroke occurred in 12 patients (1.4%). The incidence of cerebral ischemic stroke in LAA closure group was significantly lower than in non-LAA closure group (0.6% vs.2.7%, χ² = 6.452, P = 0.011).Logistic regression analysis showed that LAA closure was a significant protective factor for postoperative cerebral ischemic stroke (OR = 0.189, 95% CI: 0.039 to 0.902, P = 0.037) while history of cerebrovascular disease (OR = 4.326, 95% CI:1.074 to 17.418, P = 0.039) and preoperative diameter of left atrium (OR = 1.509, 95% CI: 1.022 to 1.098, P = 0.002) being the independent risk factors for postoperative cerebral ischemic stroke. The subgroup analysis showed that, for atrial fibrillation patients, LAA closure was a strong protective factor (OR = 0.064, 95% CI: 0.006 to 0.705, P = 0.025), but LAA closure was not a significant predictive factor (OR = 1.902, 95% CI: 0.171 to 21.191, P = 0.601) in non-atrial fibrillation patients. CONCLUSION: Concurrent LAA closure during mitral valve replacement is safe and effective to reduce the early postoperative risk of cerebral ischemic stroke in atrial fibrillation patients.

[Surgical treatment of aortic paravalvular abscess by infective endocarditis].

OBJECTIVE: To summarize the clinical features, pathology and surgical treatment experiences in the patients with aortic paravalvular abscess by infective endocarditis. METHODS: The study consisted of a retrospective analysis of 29 cases with aortic paravalvular abscess by infective endocarditis underwent surgical treatment between January 2001 and June 2013. Among the 29 patients, 22 were male and 7 were female, and the mean age was (37 ± 16) years (range from 11 to 63 years). The primary cardiac disease was congenital aortic valve malformation in 16 patients. There were 15 patients with a history of severe heart failure. Of 29 cases, 8 abscess cavities, 13 pseudoaneurysms and 6 fistulas were found, and complete aortoventricular discontinuity was present in 5 patients with serious infections. Of them, the abscess was above the annulus in 14 patients and below the annulus in 10 patients, and simultaneously involved the annulus above and below in 5 patients. 19 patients were culture positive either positive preoperative blood cultures or positive cultures of surgical specimens, including 9 patients with Staphylococcus infection. The paravalvular defect was repaired by patch in 19 cases, and by local closure in 10 cases. The valvular annulus was reconstructed simultaneously in 16 patients. Aortic valve replacement was performed in 26 patients, and Bentall procedure in 2 patients, including 23 with prosthetic mechanical valve and 5 with biological valve. RESULTS: Of the total 29 patients, 28 patients were recovered, and 1 patient was died of sepsis. During 3 months to 13 years postoperative follow-up (average 4.5 years), one was died of non-cardiac cause, and no patient had recurrent endocarditis and paravalvular leakage. CONCLUSIONS: Aortic paravalvular abscess by infective endocarditis is not uncommon, prone to heart failure. According to the different pathological manifestations, the appropriate surgical approach and strategy can achieve satisfactory outcomes.

A new strategy to stabilize the heavy metals in carbonized MSWI-fly ash using an acid-resistant oligomeric dithiocarbamate chelator.

Fly ash (FA) derived from municipal solid waste incineration (MSWI) requires safe handling before landfilling due to its extremely high salt content and the risk of leaching heavy metals (HMs) under acidic conditions. Herein, aimed at improving the acid stability of dithiocarbamates, a cost-effective oligomeric dithiocarbamate (ODTC) was developed to stabilize HMs from carbonated MSWI-FA. Spiking of 3.6 wt% ODTC reduced the HM leaching below landfill standards in China, even across the pH range of 2.0-13.0 or 8-week exposure to the natural environment. Stabilization decreased the acid-soluble/exchangeable fractions of Cd, Pb, and Zn from 22.2%, 4.49%, and 21.9% to 0.14%, 0.11%, and 12.2%, respectively, resulting in safe levels for Pb and Cd with risk assessments. Compared to DDTC and SDD, ODTC exhibited higher stability under acidic conditions after chelation with the HMs, minimized the risk of HM leaching, and significantly reduced stabilization costs. In-depth studies proved that the stabilization mechanism involved the ability of ODTC to chelate HMs strongly and form acid-resistant ODTC-HM complexes, agglomeration of the MSWI-FA grains to encapsulate the ODTC-HM complexes, transformations of the HMs from acid-soluble species to stable oxidizable and residual species, and specifically ODTC reducing high-valent Pb to more stable Pb(II) species.

Exosomes on the development and progression of renal fibrosis.

Renal fibrosis is a prevalent pathological alteration that occurs throughout the progression of primary and secondary renal disorders towards end-stage renal disease. As a complex and irreversible pathophysiological phenomenon, it includes a sequence of intricate regulatory processes at the molecular and cellular levels. Exosomes are a distinct category of extracellular vesicles that play a crucial role in facilitating intercellular communication. Multiple pathways are regulated by exosomes produced by various cell types, including tubular epithelial cells and mesenchymal stem cells, in the context of renal fibrosis. Furthermore, research has shown that exosomes present in bodily fluids, including urine and blood, may be indicators of renal fibrosis. However, the regulatory mechanism of exosomes in renal fibrosis has not been fully elucidated. This article reviewed and analysed the various mechanisms by which exosomes regulate renal fibrosis, which may provide new ideas for further study of the pathophysiological process of renal fibrosis and targeted treatment of renal fibrosis with exosomes.

Extracellular-matrix tethering regulates stem-cell fate.

To investigate how substrate properties influence stem-cell fate, we cultured single human epidermal stem cells on polydimethylsiloxane (PDMS) and polyacrylamide (PAAm) hydrogel surfaces, 0.1 kPa-2.3 MPa in stiffness, with a covalently attached collagen coating. Cell spreading and differentiation were unaffected by polydimethylsiloxane stiffness. However, cells on polyacrylamide of low elastic modulus (0.5 kPa) could not form stable focal adhesions and differentiated as a result of decreased activation of the extracellular-signal-related kinase (ERK)/mitogen-activated protein kinase (MAPK) signalling pathway. The differentiation of human mesenchymal stem cells was also unaffected by PDMS stiffness but regulated by the elastic modulus of PAAm. Dextran penetration measurements indicated that polyacrylamide substrates of low elastic modulus were more porous than stiff substrates, suggesting that the collagen anchoring points would be further apart. We then changed collagen crosslink concentration and used hydrogel-nanoparticle substrates to vary anchoring distance at constant substrate stiffness. Lower collagen anchoring density resulted in increased differentiation. We conclude that stem cells exert a mechanical force on collagen fibres and gauge the feedback to make cell-fate decisions.

Ultrasensitive refractive index sensor based on the resonant scattering effect between double air circular-holes on silicon waveguides.

Recently, we have proposed a sensitive refractive index sensor design by integrating a circular-hole defect with an etched diffraction grating (EDG) spectrometer based on amorphous silicon photonic platforms. In the present paper, we will show that a much better sensitivity (~17422 nm/RIU) can be obtained by using double circular-holes with an appropriate interval. The influence of the double-hole interval on the performance of sensing applications is also characterized. A sinusoidal pattern of the sensitivity can be found as the interval increases. However, the intensity of the resonant peak (i.e., the detectability for sensing applications) significantly oscillates as the interval varies.

Polyamide membranes with nanoscale ordered structures for fast permeation and highly selective ion-ion separation.

Fast permeation and effective solute-solute separation provide the opportunities for sustainable water treatment, but they are hindered by ineffective membranes. We present here the construction of a nanofiltration membrane with fast permeation, high rejection, and precise Cl-/SO42- separation by spatial and temporal control of interfacial polymerization via graphitic carbon nitride (g-C3N4). The g-C3N4 nanosheet binds preferentially with piperazine and tiles the water-hexane interface as revealed by molecular dynamics studies, thus lowering the diffusion rate of PIP by one order of magnitude and restricting its diffusion pathways towards the hexane phase. As a result, membranes with nanoscale ordered hollow structure are created. Transport mechanism across the structure is clarified using computational fluid dynamics simulation. Increased surface area, lower thickness, and a hollow ordered structure are identified as the key contributors to the water permeance of 105 L m2·h-1·bar-1 with a Na2SO4 rejection of 99.4% and a Cl-/SO42- selectivity of 130, which is superior to state-of-the-art NF membranes. Our approach for tuning the membrane microstructure enables the development of ultra-permeability and excellent selectivity for ion-ion separation, water purification, desalination, and organics removal.

Effect of transmembrane protein 100 on prostate cancer progression by regulating SCNN1D through the FAK/PI3K/AKT pathway.

The effects of transmembrane (TMEM) proteins in the progression of prostate cancer (PCa) remain unknown. This study aims to explore the functions of TMEM100 in PCa. To explore the expression, regulation, and effects of TMEM100 in PCa, two PCa cell lines and 30 PCa tissue samples with adjacent control tissues were examined. Online databases, immunohistochemistry, immunofluorescence, western blot, flow cytometry, colony formation, wound healing, transwell assays, and xenograft mouse models were used to explore effects of TMEM100 relevant to PCa. TMEM100 expression was shown to decrease in PCa patients, and low TMEM100 expression was associated with tumor stage and metastasis. Overexpression of TMEM100 suppressed PCa progression by inhibiting the FAK/PI3K/AKT signaling pathway. Tumor size was smaller in TMEM100 overexpressing PCa cells in xenograft mice than in control mice. We also found that TMEM100 could regulate SCNN1D by inhibiting FAK/PI3K/AKT signaling in PCa cell lines. Taken together, our findings indicate that TMEM100 is a tumor suppressor that plays a vital role in preventing PCa proliferation, migration, and invasion through inhibition of FAK/PI3K/AKT signaling. These studies suggest that TMEM100 can be used as a predictive biomarker and therapeutic target.

Classification and clinical significance of the posterior group of renal calyces.

To study the anatomical orientation of the posterior group of calyces based on reconstructed images of computerized tomography urography (CTU) and provide a novel classification with its clinical significance. Clinical data of a total of 1321 patients, who underwent CTU examination in our hospital were retrospectively analyzed. Among these, a total of 2642 3-dimensional reconstructed images of CTU scans were considered in this study. Based on the morphology of the renal calyces and the influence on the establishment of surgical access, the posterior group renal calyces are classified into 3 major types including pot-belly type, classically branched and elongated branched. The classically branched type is further classified into 3 sub-types: a, b and c, based on the association of minor calyces of the posterior group to the major calyces. Type a is derived from 1 group of major calyces only, type b is derived from 2 groups of major calyces simultaneously, and type c is derived from 3 groups of major calyces simultaneously. Statistical findings revealed that all kidneys possess posterior group calyces. The percentage of occurrence of pot-belly type, classically branched and elongated branched is 8.06%, 73.13%, and 18.81%, respectively. The anatomical typing of the classical branching type occurred in 19.36%, 68.17%, and 12.47% for types a, b, and c, respectively. In this study, the posterior group calyces were found to be present across all patients. The posterior group calyces were highest in the classical branching type, of which anatomical typing was highest in type b. The typing of the posterior group of calyces could provide an anatomical basis for percutaneous nephrolithotomy (PCNL) puncture from the posterior group.

p53 deacetylation alleviates calcium oxalate deposition-induced renal fibrosis by inhibiting ferroptosis.

Calcium oxalate (CaOx) stones are among the most common types of kidney stones and are associated with renal tubular damage, interstitial fibrosis, and chronic kidney disease. The mechanism of CaOx crystal-induced renal fibrosis remains unknown. Ferroptosis, a type of regulated cell death, is characterised by iron-dependent lipid peroxidation, and the tumour suppressor p53 is a key regulator of ferroptosis. In the present study, our results demonstrated that ferroptosis was significantly activated in patients with nephrolithiasis and hyperoxaluric mice as well as verified the protective effects of ferroptosis inhibition on CaOx crystal-induced renal fibrosis. Moreover, the single-cell sequencing database, RNA-sequencing, and western blot analysis revealed that the expression of p53 was increased in patients with chronic kidney disease and the oxalate-stimulated human renal tubular epithelial cell line, HK-2. Additionally, the acetylation of p53 was enhanced by oxalate stimulation in HK-2 cells. Mechanistically, we found that the induction of p53 deacetylation, owing to either the SRT1720-induced activation of deacetylase sirtuin 1 or the triple mutation of p53, inhibited ferroptosis and alleviated renal fibrosis caused by CaOx crystals. We conclude that ferroptosis is one of the critical mechanisms contributing to CaOx crystal-induced renal fibrosis, and the pharmacological induction of ferroptosis via sirtuin 1-mediated p53 deacetylation may be a potential target for preventing renal fibrosis in patients with nephrolithiasis.

Histone H3K27 methyltransferase EZH2 regulates apoptotic and inflammatory responses in sepsis-induced AKI.

Rationale: The role of histone methylation modifications in renal disease, particularly in sepsis-induced acute kidney injury (AKI), remains unclear. This study aims to investigate the potential involvement of the histone methyltransferase zeste homolog 2 (EZH2) in sepsis-induced AKI and its impact on apoptosis and inflammation. Methods: We first examined the expression of EZH2 in the kidney of sepsis-induced AKI (LPS injection) mice and LPS-stimulated tubular epithelial cells. We next constructed the EZH2 knockout mice to further confirm the effects of EZH2 on apoptosis and inflammatory response in AKI. And the inflammatory level of epithelial cells can be reflected by detecting chemokines and the chemotaxis of macrophages. Subsequently, we constructed the EZH2 knocked-down cells again and performed Chromatin Immunoprecipitation sequencing to screen out the target genes regulated by EZH2 and the enrichment pathway. Then we confirmed the EZH2 target gene and its regulatory pathway in vivo and in vitro experiments. Experimental results were finally confirmed using another in vivo model of sepsis-induced AKI (cecal perforation ligation). Results: The study found that EZH2 was upregulated in sepsis-induced AKI and that silencing EZH2 could reduce renal tubular injury by decreasing apoptosis and inflammatory response of tubular epithelial cells. EZH2 knockout mice showed significantly reduced renal inflammation and macrophage infiltration. Chromatin immunoprecipitation sequencing and polymerase chain reaction identified Sox9 as a target of EZH2. EZH2 was found to be enriched on the promoter of Sox9. Silencing EZH2 resulted in a significant increase in the transcriptional level of Sox9 and activation of the Wnt/ß-catenin signaling pathway. The study further reversed the effects of EZH2 silencing by silencing Sox9 or administering the Wnt/ß-catenin inhibitor icg001. It was also found that Sox9 positively regulated the expression of ß-catenin and its downstream pathway-related genes. Finally, the study showed that the EZH2 inhibitor 3-deazaneplanocin A significantly alleviated sepsis-induced AKI. Conclusion: Our results indicate that silencing EZH2 can protect renal function by relieving transcriptional inhibition of Sox9, activating the Wnt/ß-catenin pathway, and attenuating tubular epithelial apoptosis and inflammatory response of the renal interstitium. These results highlight the potential therapeutic value of targeting EZH2 in sepsis-induced AKI.

YAP/ACSL4 Pathway-Mediated Ferroptosis Promotes Renal Fibrosis in the Presence of Kidney Stones.

The potential association between calcium oxalate stones and renal fibrosis has been extensively investigated; however, the underlying mechanisms remain unclear. Ferroptosis is a novel form of cell death characterized by iron-dependent lipid peroxidation and regulated by acyl coenzyme A synthase long-chain family member 4 (ACSL4). Yes-associated protein (YAP), a transcriptional co-activator in the Hippo pathway, promotes ferroptosis by modulating ACSL4 expression. Nevertheless, the involvement of YAP-ACSL4 axis-mediated ferroptosis in calcium oxalate crystal deposition-induced renal fibrosis and its molecular mechanisms have not been elucidated. In this study, we investigated ACSL4 expression and ferroptosis activation in the kidney tissues of patients with calcium oxalate stones and in mice using single-cell sequencing, transcriptome RNA sequencing, immunohistochemical analysis, and Western blot analysis. In vivo and in vitro experiments demonstrated that inhibiting ferroptosis or ACSL4 mitigated calcium oxalate crystal-induced renal fibrosis. Furthermore, YAP expression was elevated in the kidney tissues of patients with calcium oxalate stones and in calcium oxalate crystal-stimulated human renal tubular epithelial cell lines. Mechanistically, in calcium oxalate crystal-stimulated human renal tubular epithelial cell lines, activated YAP translocated to the nucleus and enhanced ACSL4 expression, consequently inducing cellular ferroptosis. Moreover, YAP silencing suppressed ferroptosis by downregulating ACSL4 expression, thereby attenuating calcium oxalate crystal-induced renal fibrosis. Conclusively, our findings suggest that YAP-ACSL4-mediated ferroptosis represents an important mechanism underlying the induction of renal fibrosis by calcium oxalate crystal deposition. Targeting the YAP-ACSL4 axis and ferroptosis may therefore hold promise as a potential therapeutic approach for preventing renal fibrosis in patients with kidney stones.

Overexpression of sirtuin 1 attenuates calcium oxalate-induced kidney injury by promoting macrophage polarization.

Sirtuin 1 (SIRT1) protein is involved in macrophage differentiation, while NOTCH signaling affects inflammation and macrophage polarization. Inflammation and macrophage infiltration are typical processes that accompany kidney stone formation. However, the role and mechanism of SIRT1 in renal tubular epithelial cell injury caused by calcium oxalate (CaOx) deposition and the relationship between SIRT1 and the NOTCH signaling pathway in this urological disorder are unclear. This study investigated whether SIRT1 promotes macrophage polarization to inhibit CaOx crystal deposition and reduce renal tubular epithelial cell injury. Public single-cell sequencing data, RT-qPCR, immunostaining approaches, and Western blotting showed decreased SIRT1 expression in macrophages treated with CaOx or exposed to kidney stones. Macrophages overexpressing SIRT1 differentiated towards the anti-inflammatory M2 phenotype, significantly inhibiting apoptosis and alleviating injury in the kidneys of mice with hyperoxaluria. Conversely, decreased SIRT1 expression in CaOx-treated macrophages triggered Notch signaling pathway activation, promoting macrophage polarization towards the pro-inflammatory M1 phenotype. Our results suggest that SIRT1 promotes macrophage polarization towards the M2 phenotype by repressing the NOTCH signaling pathway, which reduces CaOx crystal deposition, apoptosis, and damage in the kidney. Therefore, we propose SIRT1 as a potential target for preventing disease progression in patients with kidney stones.