Multiscale X-ray phase-contrast CT unveils the evolution of bile infarct in obstructive biliary disease.

Bile infarct is a pivotal characteristic of obstructive biliary disease, but its evolution during the disease progression remains unclear. Our objective, therefore, is to explore morphological alterations of the bile infarct in the disease course by means of multiscale X-ray phase-contrast CT. Bile duct ligation is performed in mice to mimic the obstructive biliary disease. Intact liver lobes of the mice are scanned by phase-contrast CT at various resolution scales. Phase-contrast CT clearly presents three-dimensional (3D) images of the bile infarcts down to the submicron level with good correlation with histological images. The CT data illustrates that the infarct first appears on day 1 post-BDL, while a microchannel between the infarct and hepatic sinusoids is identified, the number of which increases with the disease progression. A 3D model of hepatic acinus is proposed, in which the infarct starts around the portal veins (zone I) and gradually progresses towards the central veins (zone III) during the disease process. Multiscale phase-contrast CT offers the comprehensive analysis of the evolutionary features of the bile infarct in obstructive biliary disease. During the course of the disease, the bile infarcts develop infarct-sinusoidal microchannels and gradually occupy the whole liver, promoting the disease progression.

Oregano essential oil encapsulated in zein-pectin-chitosan nanoparticles to improve the storage quality of Harbin red sausage.

In this study, the effect of oregano essential oil loaded in zein-pectin-chitosan (Zein-PC-CS-OEO) nanoparticles on the quality of Harbin red sausage during storage was examined. Zein-PC-CS-OEO nanoparticles exhibit the better encapsulation efficiency, antioxidant and antibacterial properties than these of other prepared nanoparticles, which were subsequently incorporated into Harbin red sausage with different concentrations. The physicochemical properties, bacterial community structure, and flavor characteristics of the Harbin red sausage were determined. Both thiobarbituric acid values and the growth of dominant spoilage bacteria in Harbin red sausage are inhibited by Zein-PC-CS-OEO nanoparticles, while the total aerobic bacteria count is reduced. These results indicate that the storage quality of Harbin red sausage is improved by Zein-PC-CS-OEO nanoparticles. It is worth noting that the shelf life of Harbin red sausage supplemented with 0.1 % Zein-PC-CS-OEO nanoparticles is extended to 9 d, and the flavor characteristics of which are better maintained. This study provides a new approach to extend the application of essential oil and improve the storage quality of Harbin red sausage.

BAG5 regulates HSPA8-mediated protein folding required for sperm head-tail coupling apparatus assembly.

Teratozoospermia is a significant cause of male infertility, but the pathogenic mechanism of acephalic spermatozoa syndrome (ASS), one of the most severe teratozoospermia, remains elusive. We previously reported Spermatogenesis Associated 6 (SPATA6) as the component of the sperm head-tail coupling apparatus (HTCA) required for normal assembly of the sperm head-tail conjunction, but the underlying molecular mechanism has not been explored. Here, we find that the co-chaperone protein BAG5, expressed in step 9-16 spermatids, is essential for sperm HTCA assembly. BAG5-deficient male mice show abnormal assembly of HTCA, leading to ASS and male infertility, phenocopying SPATA6-deficient mice. In vivo and in vitro experiments demonstrate that SPATA6, cargo transport-related myosin proteins (MYO5A and MYL6) and dynein proteins (DYNLT1, DCTN1, and DNAL1) are misfolded upon BAG5 depletion. Mechanistically, we find that BAG5 forms a complex with HSPA8 and promotes the folding of SPATA6 by enhancing HSPA8's affinity for substrate proteins. Collectively, our findings reveal a novel protein-regulated network in sperm formation in which BAG5 governs the assembly of the HTCA by activating the protein-folding function of HSPA8.

One-Step Microfluidic Fabrication of Bioinspired Microfibers with a Spindle-Knot Structure for Fog Harvest.

Many biomimetic microfibers have been designed from spider silk to collect water efficiently from humid air as a result of its periodic spindle-knot structure, which enhances the direct movement and convergence of captured fog droplets. Here, a hydrodynamic flow-focusing microfluidic device with a theta-shaped tube is designed for the one-step fabrication of bioinspired microfibers with a spindle-knot structure for fog harvest. The morphology of the structured microfibers, including height, width, and spacing of spindle knots, can be adjusted readily by regulating the flow rate of specific phases. The production rate of these structured microfibers can reach 1100 cm/min. Moreover, the capture, transportation, and collection performance of fog droplets on various microfibers are investigated in a fog collection platform. It is demonstrated that our one-step microfluidic device presents a ready method for the fabrication of structured microfibers with spindle knots, which provide a significant facilitation on efficient fog capture and water collection.

The Predictive Ability of Blood Neurofilament Light Chain in Predicting Cognitive Decline in the Alzheimer's Disease Continuum: A Systematic Review and Meta-Analysis.

Background: Alzheimer's disease (AD) is a neurodegenerative disease with insidious onset. Identifying candidate predictors to forecast AD dementia risk before disease onset is crucial for early diagnosis and treatment. Objective: We aimed to assess the predictive ability of blood neurofilament light (NfL) chain in anticipating cognitive decline in the AD continuum. Methods: We systematically searched PubMed, Web of Science, and Embase from inception until April 7, 2023. Longitudinal observational studies examining the association between baseline blood NfL and cognitive decline or clinical disease conversion were included based on inclusion/exclusion criteria. The final effect size was represented by adjusted hazard ratios (HR) or standardized beta (s.ß) coefficients with a 95% confidence interval (CI). Results: A total of 2,862 articles were identified, and 26 studies were included in this meta-analysis. The results indicated that baseline blood NfL could predict cognitive decline, with MMSE [s.ß= -0.17, 95% CI (-0.26, -0.07)]; PACC [s.ß= -0.09, 95% CI (-0.16, -0.03)]; ADAS-cog [s.ß= 0.21, 95% CI (0.13, 0.29)]; CDR-SOB [s.ß= 0.27, 95% CI (0.03, 0.50)]; Global cognitive composite [s.ß= -0.05, 95% CI (-0.08, -0.01)]; Memory subdomain [s.ß= -0.06, 95% CI (-0.09, -0.03)]; Language subdomain [s.ß= -0.07, 95% CI (-0.10, -0.05)]; Executive function subdomain [s.ß= -0.02, 95% CI (-0.03, -0.01)]; Visuospatial subdomain [s.ß= -0.06, 95% CI (-0.08, -0.04)]. Additionally, baseline blood NfL could predict disease progression (conversion from CU/SCD/MCI to MCI/AD) in the AD continuum [Adjust HR = 1.32, 95% CI (1.12, 1.56)]. Conclusions: Baseline blood NfL demonstrated predictive capabilities for global cognition and its memory, language, executive function, visuospatial subdomains decline in the AD continuum. Moreover, it exhibited the potential to predict disease progression in non-AD dementia participants.

hnRNPA2B1 represses the disassembly of arsenite-induced stress granules and is essential for male fertility.

Although the composition and assembly of stress granules (SGs) are well understood, the molecular mechanisms underlying SG disassembly remain unclear. Here, we identify that heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2B1) is associated with SGs and that its absence specifically enhances the disassembly of arsenite-induced SGs depending on the ubiquitination-proteasome system but not the autophagy pathway. hnRNPA2B1 interacts with many core SG proteins, including G3BP1, G3BP2, USP10, and Caprin-1; USP10 can deubiquitinate G3BP1; and hnRNPA2B1 depletion attenuates the G3BP1-USP10/Caprin-1 interaction but elevates the G3BP1 ubiquitination level under arsenite treatment. Moreover, the disease-causing mutation FUSR521C also disassembles faster from SGs in HNRNPA2B1 mutant cells. Furthermore, knockout of hnRNPA2B1 in mice leads to Sertoli cell-only syndrome (SCOS), causing complete male infertility. Consistent with this, arsenite-induced SGs disassemble faster in Hnrnpa2b1 knockout (KO) mouse Sertoli cells as well. These findings reveal the essential roles of hnRNPA2B1 in regulating SG disassembly and male mouse fertility.

Enhancing Comprehensive Performance via Capturing and Scattering the Carriers inside PESU-Based Nanocomposite Film Capacitors.

Film capacitors have become key electronic components for electrical energy storage installations and high-power electronic systems. Nonetheless, high-temperature and high-electric-field environments would cause a surge of the energy loss, placing a fundamental challenge for film capacitors applied in harsh environments. Here, we constructed a composite film, combining poly(ether sulfone) (PESU) with excellent thermal stability and large-band-gap filler boron nitride nanosheets (BNNSs). The introduction of BNNSs would form deep/shallow traps inside the dielectric polymer matrix, effectively affecting charge migration. Via density functional theory (DFT) calculation, the higher highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) energy levels of the BNNS than the matrix facilitate scattering electrons and attracting holes. The resultant composite obtains the desired discharged energy densities (Ud) of 5.89 and 3.86 J/cm3 accompanied by an efficiency above 90% at 150 and 200 °C, respectively, surpassing those of existing dielectric materials at the high-temperature conditions. The paper provides a promising composite dielectric material for high-performance film capacitors capable of operating in harsh environments.

Physiological responses of low- and high-cadmium accumulating Robinia pseudoacacia-rhizobium symbioses to cadmium stress.

The role of rhizobia in alleviating cadmium (Cd) stress in woody legumes is still unclear. Therefore, two types of black locust (Robinia pseudoacacia L.) with high and low Cd accumulation abilities were selected from 11 genotypes in China, and the effects of rhizobium (Mesorhizobium huakuii GP1T11) inoculation on the growth, CO2 and H2O gas exchange parameters, Cd accumulation, and the absorption of mineral elements of the high (SX) and low Cd-accumulator (HB) were compared. The results showed that rhizobium-inoculation significantly increased biomass, shoot Cd contents, Cd accumulation, root-to-shoot translocation factor (TF) and the absorption and accumulation of mineral elements in both SX and HB. Rhizobium-inoculation increased chlorophyll a and carotenoid contents, and the intercellular carbon dioxide concentrations in HB plants. Under Cd exposure, the high-accumulator SX exhibited a significant decrease in photosynthetic CO2 fixation (Pn) and an enhanced accumulation of Cd in leaves, but coped with Cd exposure by increasing chlorophyll synthesis, regulating stomatal aperture (Gs), controlling transpiration (Tr), and increasing the absorption and accumulation of mineral elements. In contrast, the low-accumulator HB was more sensitive to Cd exposure despite preferential accumulation of Cd in roots, with decreased chlorophyll and carotenoid contents, but significantly increased root biomass. Compared to the low-accumulator HB, non-inoculated Cd-exposed SX plants had higher chlorophyll contents, and rhizobium-inoculated Cd-exposed SX plants had higher Pn, Tr, and Gs as well as higher levels of P, K, Fe, Ca, Zn, and Cu. In conclusion, the high- and low-Cd-accumulator exhibited different physiological responses to Cd exposure. Overall, rhizobium-inoculation of black locust promoted the growth and heavy metal absorption, providing an effective strategy for the phytoremediation of heavy metal-contaminated soils by this woody legume.

Mechanisms underlying the relationship between the intersectionality of multiple social identities with depression among US adults: A population-based study on the mediating roles of lifestyle behaviors.

BACKGROUND: Despite substantial efforts to investigate the inequalities in mental health among people with marginalized identities, most research has concentrated on single social identities rather than complete persons. The current study aimed to explore the mechanisms underlying the relationship between the intersectionality of multiple social identities with depression among US adults. METHODS: Data for this study came from the National Health and Nutrition Examination Survey waves between 2015 and 2018, including a total of 11,268 US adults. A latent class analysis (LCA) was conducted to identify latent subpopulations based on sex, race/ethnicity, immigrant status, disability, household income, employment status, and education level. Interclass differences in lifestyle factors and depression were examined. Multiple mediation analysis was used to examine the mediating roles of lifestyle behaviors. RESULTS: LCA identified four potential subpopulations: "least marginalized", "immigrant minorities", "disabled, less-educated non-workers", and "low-income minorities" groups. There was enormous heterogeneity in mental health among immigrant minority individuals. The "disabled, less-educated non-workers" group had the highest rates of depression; in contrast, the "immigrant minorities" group had the best mental health, even better than that of the "least marginalized" group. Distributions of lifestyle factors followed a similar pattern. In addition, lifestyle behaviors significantly mediated the relationship between intersectional social identities and depression. LIMITATIONS: The cross-sectional design prevented establishment of the causality of relationships. CONCLUSIONS: This study suggests that applying a person-centered approach is important when examining intersectional inequalities in mental health and highlights the effects of structural social hierarchies on individuals' health behaviors and mental health.

Quantitative assessment of bone marrow infiltration and characterization of tumor burden using dual-layer spectral CT in patients with multiple myeloma.

BACKGROUND: The aim of the study was to evaluate whether virtual calcium subtraction (VNCa) image extracted from dual-layer spectral CT could estimate bone marrow (BM) infiltration with MRI as the reference standard and characterize tumor burden in patients with multiple myeloma (MM). PATIENTS AND METHODS: Forty-seven patients with newly diagnosed MM were retrospectively enrolled. They had undergone whole-body low-dose dual-layer spectral CT (DLCT) and whole-body MRI within one week. VNCa images with calcium-suppressed (CaSupp) indices ranging from 25 to 95 at an interval of 10 and apparent diffusion coefficient (ADC) maps were quantitatively analyzed on vertebral bodies L1-L5 at the central slice of images. The optimal combination was selected by correlation analysis between CT numbers and ADC values. Then, it was used to characterize tumor burden by correlation analysis and receiver operating characteristic (ROC) curves analysis, including plasma cell infiltration rate (PCIR), high serum-free light chains (SFLC) ratio and the high-risk cytogenetic (HRC) status. RESULTS: The most significant quantitative correlation between CT numbers of VNCa images and ADC values could be found at CaSupp index 85 for averaged L1-L5 (r = 0.612, p < 0.001). It allowed quantitative evaluation of PCIR (r = 0.835, p < 0.001). It could also anticipate high SFLC ratio and the HRC status with area under the curve (AUC) of 0.876 and 0.760, respectively. CONCLUSIONS: The VNCa measurements of averaged L1-L5 showed the highest correlation with ADC at CaSupp index 85. It could therefore be used as additional imaging biomarker for non-invasive assessment of tumor burden if ADC is not feasible.

Contrast Agent Reflux in Transvaginal 4-D Hysterosalpingo-Contrast Sonography: Influencing Factors and Coping Strategies.

ABSTRACT: Transvaginal 4-D hysterosalpingo-contrast sonography (TV 4-D HyCoSy) plays an important role in the detection and diagnosis of clinical female infertility. The purposes of this study were to analyze the influencing factors of TV 4murD HyCoSy complicated with contrast agent reflux and to provide evidence for clinical diagnosis and treatment. Female patients diagnosed as infertility by transvaginal hysterosalpingography from January 2021 to December 2022 were included. The characteristics of patients with and without contrast agent reflux were evaluated. Pearson correlation and logistic regression were conducted to analyze the related factors affecting the occurrence of contrast reflux. A total of 416 patients undergoing TV 4-D HyCoSy were included, and the incidence of contrast agent reflux in patients undergoing TV 4-D HyCoSy was 38.94%. Pearson correlation analysis results indicated that history of uterine cavity operation ( r = 0.556), adenomyosis of uterus ( r = 0.584), examination on less than 5 days after menstruation ( r = 0.602), endometrial thickness ( r = 0.566), and endometrial polyps ( r = 0.575) are all correlated with contrast agent reflux in patients undergoing 4-D HyCoSy (all P < 0.05). Logistic regression analysis showed that history of uterine cavity operation (odds ratio [OR], 1.109; 95% confidence interval [CI], 1.012-1.872), adenomyosis of uterus (OR, 2.026; 95% CI, 1.864-2.425), examination on less than 5 days after menstruation (OR, 2.465; 95% CI, 2.118-2.851), endometrial thickness less than 6 mm (OR, 2.866; 95% CI, 2.095-2.957), and endometrial polyps (OR, 1.587; 95% CI, 1.137-1.744) were the influencing factors of contrast agent reflux in patients undergoing (all P < 0.05). The incidence of contrast agent reflux in TV 4-D HyCoSy is high, and there are many influencing factors. Clinical medical workers should take early measures based on these influencing factors to reduce the contrast agent reflux.

A flame-retardant and conductive fabric-based triboelectric nanogenerator: Application in fire alarm and emergency evacuation.

The fabrics commonly used in architectural decorative materials pose significant fire hazards due to their flammability and rapid fire spread. Moreover, the traditional fire-alarm systems may fail to function properly in complex fire environments owing to power supply disruptions. In this study, we developed a low-cost and eco-friendly flame-retardant conductive fabric-based triboelectric nanogenerator (FCF-TENG) by integrating flame-retardant conductive nylon fabric and polytetrafluoroethylene soaked cotton fabric. This nanogenerator exhibits excellent flame-retardant properties and remarkable energy-harvesting capabilities. The nylon fabric, treated with layer-by-layer self-assembly method, possesses outstanding self-extinguishing capability and melt-dripping resistance. Additionally, the electrical performance of FCF-TENG significantly improves, with a 10-fold boost in conductivity, and the open-circuit voltage increases by 84% to 92 V. Besides, by incorporating the rectifier circuit, the FCF-TENG is capable of completely charging a 1 µF capacitor within 30 s. Furthermore, the FCF-TENG was successfully applied as a self-powered sensor in the fire-alarm system and served as a safety exit indicator for evacuees and fire rescue. This work presents an effective and innovative application of multifunctional smart textiles for energy harvesting and self-powered sensing.

BLK positively regulates TLR/IL-1R signaling by catalyzing TOLLIP phosphorylation.

TLR/IL-1R signaling plays a critical role in sensing various harmful foreign pathogens and mounting efficient innate and adaptive immune responses, and it is tightly controlled by intracellular regulators at multiple levels. In particular, TOLLIP forms a constitutive complex with IRAK1 and sequesters it in the cytosol to maintain the kinase in an inactive conformation under unstimulated conditions. However, the underlying mechanisms by which IRAK1 dissociates from TOLLIP to activate TLR/IL-1R signaling remain obscure. Herein, we show that BLK positively regulates TLR/IL-1R-mediated inflammatory response. BLK-deficient mice produce less inflammatory cytokines and are more resistant to death upon IL-1ß challenge. Mechanistically, BLK is preassociated with IL1R1 and IL1RAcP in resting cells. IL-1ß stimulation induces heterodimerization of IL1R1 and IL1RAcP, which further triggers BLK autophosphorylation at Y309. Activated BLK directly phosphorylates TOLLIP at Y76/86/152 and further promotes TOLLIP dissociation from IRAK1, thereby facilitating TLR/IL-1R-mediated signal transduction. Overall, these findings highlight the importance of BLK as an active regulatory component in TLR/IL-1R signaling.

Repopulating Kupffer cells originate directly from hematopoietic stem cells.

BACKGROUND: Kupffer cells (KCs) originate from yolk-sac progenitors before birth. Throughout adulthood, they self-maintain independently from the input of circulating monocytes (MOs) at a steady state and are replenished within 2 weeks after having been depleted, but the origin of repopulating KCs in adults remains unclear. The current paradigm dictates that repopulating KCs originate from preexisting KCs or monocytes, but there remains a lack of fate-mapping evidence. METHODS: We first traced the fate of preexisting KCs and that of monocytic cells with tissue-resident macrophage-specific and monocytic cell-specific fate-mapping mouse models, respectively. Secondly, we performed genetic lineage tracing to determine the type of progenitor cells involved in response to KC-depletion in mice. Finally, we traced the fate of hematopoietic stem cells (HSCs) in an HSC-specific fate-mapping mouse model, in the context of chronic liver inflammation induced by repeated carbon tetrachloride treatment. RESULTS: By using fate-mapping mouse models, we found no evidence that repopulating KCs originate from preexisting KCs or MOs and found that in response to KC-depletion, HSCs proliferated in the bone marrow, mobilized into the blood, adoptively transferred into the liver and differentiated into KCs. Then, in the chronic liver inflammation context, we confirmed that repopulating KCs originated directly from HSCs. CONCLUSION: Taken together, these findings provided in vivo fate-mapping evidence that repopulating KCs originate directly from HSCs, which presents a completely novel understanding of the cellular origin of repopulating KCs and shedding light on the divergent roles of KCs in liver homeostasis and diseases.

Associations of Lifestyle Patterns With Overweight and Depressive Symptoms Among United States Emerging Adults With Different Employment Statuses.

Objective: To identify lifestyle patterns in emerging adults and examine the association of lifestyle patterns with overweight and depression. Methods: Data was from the National Health and Nutrition Examination Survey between 2011 and 2018. A latent class analysis (LCA) was conducted with 2,268 US emerging adults based on sedentary behavior, moderate-to-vigorous physical activity, diet, sleep, alcohol drinking, and cigarette smoking. The associations of lifestyle groups with overweight and depression were examined by logistic regression and were further stratified by employment status. Results: The LCA results favored a four-class solution: "unhealthy but non-substance use" (59%), "healthy but sleepless and drinking" (12%), "unhealthy lifestyle" (15%), and "healthy but sedentary" group (14%). Compared to the "unhealthy lifestyle" group, participants in the "unhealthy but non-substance use" (coef. = -1.44, SE = 0.27; OR = 0.40, 95% CI [0.26, 0.61]), "healthy but sleepless and drinking" (coef. = -1.49, SE = 0.35; OR = 0.38, 95% CI [0.20, 0.72]), and "healthy but sedentary" (coef. = -1.97, SE = 0.34; OR = 0.29, 95% CI [0.14, 0.57]) groups had lower depression severity and reported fewer depressive symptoms. Moreover, lifestyle groups and health outcomes showed different relationships among employed and unemployed participants. Conclusion: This study found that the combinations of lifestyle behaviors had synergistic effects on mental health, and such effects differed by employment status.

Tyrosine phosphorylation of IRF3 by BLK facilitates its sufficient activation and innate antiviral response.

Viral infection triggers the activation of transcription factor IRF3, and its activity is precisely regulated for robust antiviral immune response and effective pathogen clearance. However, how full activation of IRF3 is achieved has not been well defined. Herein, we identified BLK as a key kinase that positively modulates IRF3-dependent signaling cascades and executes a pre-eminent antiviral effect. BLK deficiency attenuates RNA or DNA virus-induced ISRE activation, interferon production and the cellular antiviral response in human and murine cells, whereas overexpression of BLK has the opposite effects. BLK-deficient mice exhibit lower serum cytokine levels and higher lethality after VSV infection. Moreover, BLK deficiency impairs the secretion of downstream antiviral cytokines and promotes Senecavirus A (SVA) proliferation, thereby supporting SVA-induced oncolysis in an in vivo xenograft tumor model. Mechanistically, viral infection triggers BLK autophosphorylation at tyrosine 309. Subsequently, activated BLK directly binds and phosphorylates IRF3 at tyrosine 107, which further promotes TBK1-induced IRF3 S386 and S396 phosphorylation, facilitating sufficient IRF3 activation and downstream antiviral response. Collectively, our findings suggest that targeting BLK enhances viral clearance via specifically regulating IRF3 phosphorylation by a previously undefined mechanism.

Touch, press and stroke: a soft capacitive sensor skin.

Soft sensors that can discriminate shear and normal force could help provide machines the fine control desirable for safe and effective physical interactions with people. A capacitive sensor is made for this purpose, composed of patterned elastomer and containing both fixed and sliding pillars that allow the sensor to deform and buckle, much like skin itself. The sensor differentiates between simultaneously applied normal force and shear using summation and differences of signals from four deformable capacitors. Cross talk from shear to normal force is less than 2.5%, and between shear axes is less than 10%. Normal and shear stress sensitivity is 0.49 kPa and 0.31 kPa respectively, with a minimum displacement resolution of 40 µm. In addition, finger proximity is detectable at a range of up to 15 mm. The operation is demonstrated on a simple gripper holding a cup. The combination of features and the straightforward fabrication method make this sensor a candidate for implementation as a sensing skin for humanoid robotics applications.

Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2-dependent mechanism.

BACKGROUND: N6 -methyladenosine (m6A) is of great importance in renal physiology and disease progression, but its function and mechanism in renal fibrosis remain to be comprehensively and extensively explored. Hence, this study will explore the function and potential mechanism of critical regulator-mediated m6A modification during renal fibrosis and thereby explore promising anti-renal fibrosis agents. METHODS: Renal tissues from humans and mice as well as HK-2 cells were used as research subjects. The profiles of m6A modification and regulators in renal fibrosis were analysed at the protein and RNA levels using Western blotting, quantitative real-time polymerase chain reaction and other methods. Methylation RNA immunoprecipitation sequencing and RNA sequencing coupled with methyltransferase-like 3 (METTL3) conditional knockout were used to explore the function of METTL3 and potential targets. Gene silencing and overexpression combined with RNA immunoprecipitation were performed to investigate the underlying mechanism by which METTL3 regulates the Ena/VASP-like (EVL) m6A modification that promotes renal fibrosis. Molecular docking and virtual screening with in vitro and in vivo experiments were applied to screen promising traditional Chinese medicine (TCM) monomers and explore their mechanism of regulating the METTL3/EVL m6A axis and anti-renal fibrosis. RESULTS: METTL3 and m6A modifications were hyperactivated in both the tubular region of fibrotic kidneys and HK-2 cells. Upregulated METTL3 enhanced the m6A modification of EVL mRNA to improve its stability and expression in an insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2)-dependent manner. Highly expressed EVL binding to Smad7 abrogated the Smad7-induced suppression of transforming growth factor-ß (TGF-ß1)/Smad3 signal transduction, which conversely facilitated renal fibrosis progression. Molecular docking and virtual screening based on the structure of METTL3 identified a TCM monomer named isoforsythiaside, which inhibited METTL3 activity together with the METTL3/EVL m6A axis to exert anti-renal fibrosis effects. CONCLUSIONS: Collectively, the overactivated METTL3/EVL m6A axis is a potential target for renal fibrosis therapy, and the pharmacological inhibition of METTL3 activity by isoforsythiaside suggests that it is a promising anti-renal fibrosis agent.

Cold stress causes liver damage by inducing ferroptosis through the p38 MAPK/Drp1 pathway.

Acute extreme cold exposure impairs human health and even causes hypothermia which threatens human life. Liver, as a hub in metabolism and thermogenesis, is vital for cold acclimatization. Although accumulating evidence has suggested that cold exposure can cause liver damage, the underlying mechanisms remain poorly understood. This study investigated the role and underlying mechanisms of ferroptosis in cold stress-induced liver damage. To evaluate the role of ferroptosis in cold stress-induced liver damage, rats were pretreated with ferroptosis inhibitor liproxstatin-1 (Lip-1) before exposed to -10 °C for 8 h. Core body temperature was recorded. The levels of ferroptosis-related indicators were examined with the corresponding assay kits or by western blotting. Hepatic pathological changes were analyzed by hematoxylin-eosin staining and ultrastructural observation. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured to assess liver function. Rats were also pretreated with p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 or Dynamin-related protein 1 (Drp1) inhibitor Mdivi-1 to determine the underlying mechanisms. We found that Lip-1 inhibited ferroptosis, attenuated hepatic pathological damages and blocked the increased ALT and AST levels in cold-exposed rats. Moreover, Mdivi-1 inhibited mitochondrial fission and suppressed ferroptosis. Furthermore, SB203580 and Mdivi-1 administration alleviated cold stress-induced liver injury. Our results suggested that cold stress caused liver damage partially by inducing ferroptosis through the p38 MAPK/Drp1 pathway. These findings might provide an effective preventive and therapeutic target for cold stress-induced liver injury.