Electron Microscopy Research Articles

Gold-coated Impedance Biosensors on PCB and PET for Real-Time Monitoring of Cancer Cells

Abstract Electrical cell-substrate impedance sensing (ECIS) biosensors are widely used for in vitro cancer cell monitoring as they are label-free, require small sample volumes, and allow real-time monitoring. ECIS electrodes are typically made of pure gold, but the usage of pure gold electrodes is too costly for single-use applications. As an alternative, this work proposes the use of gold coatings on a printed sensor's electrodes. The interdigitated electrode design was used on glass fiber-reinforced epoxy resin for printed circuit boards (PCB), and polyethylene terephthalate (PET). The Cu/Ni electrodes on PCB were electroplated with Au, while the Cu/Ni electrodes on PET were coated with Au using an electroless technique. The physicochemical properties were studied using optical microscopy, scanning electron microscopy, and energy-dispersive spectroscopy. Electrochemical characterization was done using cyclic voltammetry and electrochemical impedance spectroscopy. Biocompatibility assessment and sensor functionality tests were done by culturing SiHa cervical cancer cell lines on these sensors and impedance measurements. The results show that both electroplated and electroless sensors were biocompatible and suitable to monitor SiHa cell growth. Electrochemical migration effect was observed on the sensors where the reaction occurred at 1.2V DC for the PCB sensor and 1.0V DC for the PET sensor.

Characterization and genome analysis of Klebsiella phages with lytic activity against Klebsiella pneumoniae.

Klebsiella pneumoniae is an important gram-negative opportunistic pathogen that causes a variety of infectious diseases. As K. pneumoniae are becoming increasingly resistant to antibiotics, the use of bacteriophages may offer a non-antibiotic-based approach to treat these infections. In the present study, five lytic bacteriophages, 2044307w, k2044hw, k2044ew, k2044302 and 2146hw specific to K. pneumoniae were isolated from hospital sewage and characterized. They belong to group A of the KP32viruses based on transmission electron microscopy (TEM) and genome analysis. These bacteriophages have an extremely narrow host spectrum. The phenotypic characteristics of the phages were determined using lysis assay, pH, and temperature stability tests. This contributes to expanding our understanding of K. pneumoniae phages.

Substantive Dimethicone-Based Mucoadhesive Coatings

It is challenging to deliver therapeutics in the oral environment due to the wet surfaces, the nature of the mucosa and the potential for saliva washout. In this study, the development of a mucoadhesive dimethicone-based oral carrier system for adhesion to the hard tissue and mucosa in the mouth was examined. This study reports the viscosity and mucoadhesion of dimethicone based polymer blends. The viscosity of the materials was measured using a rheometer. The mucoadhesion of these materials was determined as the work of adhesion and peak tack force using the tensile test method with a texture analyzer. Materials were prepared with either calcium and phosphate salts or sodium fluoride as potential therapeutics for promoting remineralization and treating dentin hypersensitivity by mechanical occlusion. Scanning electron microscopy was used to look at mineral deposition on the surface of dental hard tissue after the application of the dimethicone-based formulations. The results of this study confirm the potential for using these dimethicone-based materials as mucoadhesive therapeutic delivery systems in the oral environment.

Enhancing clay soil reinforcement using MICP-BIN method with biochar-induced nucleation

The microbially induced carbonate precipitation (MICP) method has gained extensive use in the realm of soil stabilisation. This study presents a novel microbial solidification technique known as the biochar-induced nucleation (MICP-BIN) method for reinforcing clay soil. Corn stover biochar was employed as an auxiliary material, mixed with dry clay powder, and subsequently reinforced using the MICP technique. Direct shear tests, drying experiments, X-ray diffraction, and scanning electron microscopy were also conducted to investigate the efficacy of the MICP-BIN method. Compared with that of the remoulded soil, the shear strength of the treated soil exhibited a substantial increase of 22.4%, the CaCO3 content increased by 30%, and the shear strength improved by 389.5%. Similarly, the internal friction angle exhibited increases of 22.7% and 405.2%, while the cohesion showed improvements of 9.4% and 344.3%, respectively. The MICP-BIN method is found to increase suction for a given water content. Furthermore, biochar-amended soil possesses highest water-retention ability (especially at lower suction magnitude), followed by MICP-amended soil, MICP-BIN-amended soil, and untreated soil. This study demonstrated the promising results achieved by combining biochar and the MICP-BIN technique, providing a novel approach for reinforcing soft ground.

Tailoring the Lithium Deposition on Cu Substrate by a Functionalized‐polysiloxane Layer

Owing to the high theoretical capacity of 3860 mAh·g‐1 and low redox potential, lithium metal is the best candidate for the development of next generation high‐energy density lithium‐ion batteries. However, notorious lithium dendrites growth and the poor compatibility of liquid electrolyte hinder the commercialization of lithium metal batteries. In this work, an anode‐less system was used to understand the change in lithium deposition on Cu current collector after the additional functionalized‐polysiloxane (PE) layer. The PE structure consists of lithiophobic and lithiophilic side chains which facilitate the uniform lithium deposition on Cu substrate. This evidence was collected by scanning electron microscopy (SEM) after the cycling test of half‐cell configuration and the lithium deposition with different current densities. The reversibility was improved by 5% compared with the bare Cu. In addition, the potential polarization was lowered after the addition of PE layer on bare Cu. Thus, the higher cycle stability (40%) and more stable coulombic efficiency are observed on the PE@Cu||NCM83 cell compared with the bare‐Cu||NCM83 during the cycle test.

Effect of silver nanoparticles in polyethylene packaging on physicochemical, microbiological, and textural properties of oil cake.

This study investigated silver nanoparticle (SNP)/titanium dioxide polyethylene films as chemical preservative substitutes to increase the shelf life of oil cake. The research examined three types of packaging films: standard low-density polyethylene (LDPE), LDPE with 3% SNP, and LDPE with 5% SNP, and assessed their impact on oil cake shelf life. Analysis of packaging films included scanning and transmission electron microscopy (SEM and TEM), and Fourier-transform infrared spectroscopy. The study evaluated the effects of SNP on chemical and microbial stability on oil cakes in different storage days (1st, 15th, 30th, and 37th days) at 25 °C. Parameters such as moisture content, water vapor permeability, water activity, peroxide, fat acidity, texture, and microorganisms were studied. The results demonstrated that SNP-containing films can extend the shelf life of oil cake up to 37 days, marked by a significant decrease in microbial growth, particularly mold, compared to conventional packaging films. Notably, films with 5% SNP concentrations exhibited the highest efficacy in preserving the quality of the oil cake. Higher concentrations of SNP led to an increased reduction in microbial load and enhanced mold control while maintaining favorable quality assessment indices throughout the storage period from day 1 to day 30. Remarkably, oil cake packaged with a 5% SNP polymer film displayed the most promising outcomes, extending shelf life up to 30 days at 25 °C.

Restructuring the Basic Design of Several Accelerator-Based Concrete Mixes by Integrating Superplasticizers

The increasing demand for infrastructure, the need to consolidate aging structures, and the effects of climate change imply the replacement or improvement of traditional concrete. This study investigates three accelerators and their mixtures (Ca(NO3)2·4H2O, Al2(SO4)3·18H2O, and Na2S2O3·5H2O) (series I) and their counterparts with superplasticizers (Dynamon SR41) (series II) as additives in standard concrete to improve its functionality. The standard concrete and new concrete mixes were analyzed using X-ray diffraction (XRD), scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX), and tests for water absorption, bulk density, and compressive strength. XRD analysis showed that all concrete mixes had similar structures composed of quartz, portlandite, larnite, calcium silicate, ettringite, albite, and muscovite in varying proportions. Their microstructures, as shown by SEM images, revealed the presence of ettringite, portlandite, and C-S-H gel at high magnification (1–5 kx). The addition of the superplasticizer remodeled the surface of the concrete mix, reducing the pore radius and increasing its compaction. These changes helped to reduce its bulk density while increasing the compressive strength. The results showed that all the concrete mixtures are similar to the standard concrete and can replace it for better functionality, but Na2S2O3·5H2O with superplasticizer concrete mixture had the higher compressive strength, supplying additional benefits.

Tetramethylpyrazine inhibits ferroptosis in spinal cord injury by regulating iron metabolism through the NRF2/ARE pathway

BackgroundTetramethylpyrazine (TMP) is a natural alkaloid compound with antioxidant and neuroprotective effects. We hypothesized that TMP could exert neuroprotective effects by inhibiting ferroptosis through modulating iron metabolism, but its mechanism is unclear. Through in vivo and in vitro experiments, we have explored how TMP can regulate neurons’ iron metabolism through the NRF2/ARE pathway to Inhibit ferroptosis.MethodsIn the in vivo experiment, the effects of TMP on nerve function and secondary spinal cord injury were observed through behavioral tests and morphology staining. Transmission electron microscopy, molecular biology tests and immunofluorescence staining were used to investigate the role of TMP in the regulation of iron metabolism and ferroptosis through the Nrf2/ARE pathway. Using in vitro experiments to investigate the mechanism of TMP in inhibiting ferroptosis through the Nrf2/ARE pathway.ResultsFirstly, through in vivo experiments, we found that TMP improves motor function of rats with spinal cord injury, reduces spinal cord tissue damage and nerve cell death caused by secondary injury. Moreover, neuronal death and the formation of spinal cord cavities are inhibited by TMP. By regulating lipid peroxidation, TMP can inhibit mitochondrial damage and reduce ROS accumulation. Our study also demonstrated that TMP regulates iron metabolism through the NRF2/ARE pathway to inhibit ferroptosis and repair spinal cord injury. To further explore the regulatory mechanisms of TMP we down-regulating Nrf2 expression in subsequent in vitro experiments. We find that a key ferroptosis pathway, lipid peroxidation, can be regulated by TMP. Additionally, TMP inhibits iron overload-mediated ferroptosis by increasing Nrf2 transcriptional activity.ConclusionA regulatory effect of TMP on the NRF2/ARE pathway was found in both in vitro and in vivo experiments. It promotes the transcription and translation of iron metabolizing and antioxidant molecules. Our study explored the inhibitory effect of TMP on ferroptosis from the iron metabolism pathway and provided new ideas for the treatment of SCI.

Magnetic hydrogel scaffold based on hyaluronic acid/chitosan and gelatin natural polymers.

Owing to their native extracellular matrix-like features, magnetic hydrogels have been proven to be promising biomaterials as tissue engineering templates In the present work, magnetic hydrogels scaffold based on chitosan, gelatin, hyaluronic acid, containing Fe3O4 as magnetic nanoparticles (IONPs) were prepared. The prepared hydrogels were loaded with ciprofloxacin hydrochloride as a model drug. The magnetic hydrogel was prepared using different volumes of chitosan, 1%, gelatin, 10%, and hyaluronic acid, 1% in glutaraldehyde as the crosslinking agent and Fe3O4 as magnetic nanoparticles. The hydrogel scaffold and magnetic scaffold hydrogel samples were characterized by scanning electron microscopy (SEM), vibrating sample magnetometry (VSM), and Fourier-transform infrared spectroscopy (FTIR). The porosity, mechanical properties, swelling degree, and antibacterial activity of the hydrogel scaffold were also determined as well as the drug release profiles of the hydrogels. SEM imaging revealed that the magnetic hydrogel scaffold showed a relatively rough morphology with an irregular surface. The data obtained indicated that the hydrogel surface has three-dimensional porous microstructures and the porosity varied depending on the hydrogel formulation. The breaking load of the hydrogel scaffold increased from 1.361 Kgf to 4.98 Kgf by increasing the glutaraldehyde concentration from 0.2 mL to 0.8 mL. Swelling degree values in water were from 250 to 2000% after 24h. The antibacterial activity of the hydrogel scaffold ranged from 54% to about 97% for Gram-positive bacteria (S. aureus) and from about 26-92% for Gram-negative bacteria (E. coli). The ciprofloxacin hydrochloride loaded hydrogel has a sustained release of ciprofloxacin hydrochloride over 10h. The presence of IONPs gave a faster release of ciprofloxacin hydrochloride.

Structure-dynamics relation in metallic glass revealed by 5-dimensional scanning transmission electron microscopy

Structure-dynamics relation in metallic glass revealed by 5-dimensional scanning transmission electron microscopy

Ethanol extract of cassia seed alleviates metabolic dysfunction-associated steatotic liver disease by acting on multiple lipid metabolism-related pathways

Abstract Background and objective In northern China’s cold regions, the prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) exceeds 50%, significantly higher than the national and global rates. MASLD is an important risk factor for cardiovascular and cerebrovascular diseases, including coronary heart disease, stroke, and tumors, with no specific therapeutic drugs currently available. The ethanol extract of cassia seed (CSEE) has shown promise in lowering blood lipids and improving hepatic steatosis, but its mechanism in treating MASLD remains underexplored. This study aims to investigate the therapeutic effects and mechanisms of CSEE. Methods MASLD models were established in male Wistar rats and golden hamsters using a high fat diet (HFD). CSEE (10, 50, 250 mg/kg) was administered via gavage for six weeks. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), as well as liver TC and TG, were measured using biochemical kits. Histopathological changes in the liver were evaluated using Oil Red O staining, Hematoxylin-eosin (H&E) staining, and transmission electron microscopy (TEM). HepG2 cell viability was assessed using the cell counting kit-8 (CCK8) and Calcein-AM/PI staining. Network pharmacology was used to analyze drug-disease targets, and western blotting was used to confirm these predictions. Results CSEE treatment significantly reduced serum levels of TC, TG, LDL-C, ALT, and AST, and improved liver weight, liver index, and hepatic lipid deposition in rats and golden hamsters. In addition, CSEE alleviated free fatty acid (FFA)-induced lipid deposition in HepG2 cells. Molecular biology experiments demonstrated that CSEE increased the protein levels of p-AMPK, p-ACC, PPARα, CPT1A, PI3K P110 and p-AKT, while decreasing the protein levels of SREBP1, FASN, C/EBPα, and PPARγ, thus improving hepatic lipid metabolism and reducing lipid deposition. The beneficial effects of CSEE were reversed by small molecule inhibitors of the signaling pathways in vitro. Conclusion CSEE improves liver lipid metabolism and reduces lipid droplet deposition in Wistar rats and golden hamsters with MASLD by activating hepatic AMPK, PPARα, and PI3K/AKT signaling pathways.

LCN2 aggravates diabetic cataracts by promoting ferroptosis in lens epithelial cells

Abstract Background Cataracts are the leading cause of reversible blindness worldwide. Diabetic cataract (DC), a prevalent complication of diabetes mellitus, is characterized by its high occurrence, rapid progression, and severe impact. The prevalence of diabetes varies greatly between the northern and southern regions, with higher rates observed among northern residents. DC-induced lens opacity is mainly attributed to oxidative stress. However, it remains unclear whether ferroptosis, a form of regulated cell death, occurs in crystalline epithelial cells during the pathogenesis, which may represent a novel mechanism contributing to DC. Methods Transmission electron microscopy, quantitative assays for iron levels and reactive oxygen species (ROS), real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, immunofluorescence, and immunohistochemistry were used to detect ferroptosis. Gene editing techniques were utilized to study the regulatory relationships among lipocalin 2 (LCN2), glutathione peroxidase 4 (GPX4), and ferritin heavy chain (FTH). Local knockdown of the LCN2 gene in B-3 cells and the eyes of Sprague Dawley (SD) rats was performed to verify and further explore the role and regulatory mechanisms of LCN2 in DC-associated ferroptosis. Results An in vitro model using high glucose levels and an in vivo model with streptozotocin-induced diabetes in SD rats were successfully established. Ferroptosis was observed in both in vitro and in vivo experiments. LCN2 protein was normally expressed in human and rat lens epithelial cells, but its expression significantly increased during ferroptosis. The ferroptosis inhibitor, ferrostatin-1 (Fer-1) effectively inhibited ferroptosis and reduced LCN2 protein expression. Notably, local knockdown of LCN2 via gene editing protected lens epithelial cells from ferroptosis in vitro and slowed the progression of DC in SD rats in vivo. Conclusion Our findings underscore the significant role of ferroptosis in the pathogenesis of DC, suggesting that selectively targeting LCN2 activation and enhancing ferroptosis resistance may offer a novel therapeutic approach for treating DC.

Effect of carbon nanotubes and zinc oxide on electrical and mechanical properties of polyvinyl alcohol matrix composite by electrospinning method.

In this study, polymer composite nanofibers and thin membranes were synthesized using Carbon Nanotubes (CNTs) and Zinc Oxide (ZnO) as fillers in a Polyvinyl Alcohol (PVA) matrix, aiming to evaluate their electrical and mechanical properties. The composite nanofibers and thin membranes were prepared by incorporating different weight ratios of CNTs and ZnO into the PVA matrix using electrospinning and solution casting techniques, respectively. Solutions were prepared by mixing specific weight ratios of PVA, ZnO, and CNTs, followed by magnetic stirring and ultrasonication for homogenization. Electrospinning was performed at 20kV with a syringe-to-collector distance of 14.5cm at flow rate of 2.4ml/hr. The composites were characterized using FTIR spectroscopy and Scanning Electron Microscopy (SEM). The PVA/5%ZnO/0.5%CNT composite exhibited the highest dielectric constant of 10.3 at high frequencies, while PVA/5%CNT showed the highest capacitance of 31.1 pF at 2MHz. The maximum AC conductivity of 2.72 × 10- 7S/m was also observed for the PVA/5%ZnO/0.5%CNT composite. Mechanical testing revealed significant improvements in Young's modulus, stress yield, and load yield, with PVA/5%CNT achieving a Young's modulus of 387.12 MPa and a stress yield of 6.92 MPa. The addition of ZnO and CNT fillers resulted in enhanced electrical and mechanical properties, making these composites suitable for applications in microelectronic devices and packaging materials.

Visualizing supramolecular assembly behavior, stimulus response, and solid state emission of higher-order Pt2+ aggregates.

Here, we present the first instance of a highly efficient red tetramer aggregate with tunable emission based on a cationic platinum(II) complex in conjunction with a silver cluster anion counterpart. This system exhibits multicolor emission response behaviors, which can be conveniently and directly detected through spectroscopic analysis, showcasing intriguing luminescence changes. The self-assembly of Pt⋯, π-π, and hydrogen bonding interactions not only enables an intriguing color adjustment from green to yellow emission, and eventually to red emission, but also demonstrates the co-existence of the monomer, excimer, and aggregation. These phenomena are further accompanied by well-defined nanostructures. The self-assembly process of these structures exhibits an isodesmic growth mechanism, which is dependent on temperature. In this regard, it exhibits potential applicability in multi-mode logic gates that rely on external stimuli such as concentration, solvent, and temperature. The sensitivity of the aggregates towards chemical stimuli combined with their exceptionally bright emission characteristics renders them suitable for diverse applications including solid-state lighting sensing mechanisms and anticounterfeiting measures. The multi-stimuli responsive phosphorescence and self-assembly behaviors of the cationic platinum(II) complex were substantiated by X-ray crystal structure determination, 1H NMR analysis spectroscopic investigations, computational calculations and scanning electron microscopy (SEM) studies.

Mechanisms of ROS-induced mitochondria-dependent apoptosis in Phaeodactylum tricornutum under AgNPs exposure

IntroductionNanomaterials such as silver nanoparticles (AgNPs) have gained widespread application across various fields. However, the large-scale production and application of AgNPs have raised concerns about their distribution in the environment and potential pollution issues.MethodsThis study investigates the toxic effects of AgNPs on the diatom Phaeodactylum tricornutum by employing electron microscopy for cellular observation, quantifying apoptotic cell numbers, and measuring antioxidant indicators. The research examines how varying concentrations of AgNPs induce stress in P. tricornutum and the specific mechanisms of the toxic effect.ResultsThe findings reveal that AgNPs induce apoptosis in P. tricornutum cells by triggering a mitochondria-mediated pathway, marked by a decrease in mitochondrial membrane potential and the activation of caspase enzymes. Additionally, AgNP exposure results in an overproduction of reactive oxygen species (ROS) within the algal cells, leading to lipid peroxidation of the cell membrane and a consequent increase in malondialdehyde (MDA) levels. This oxidative stress response induces the upregulation of antioxidant enzyme activities in an attempt to mitigate the excessive ROS.DiscussionROS is identified as the primary factor responsible for inducing mitochondria-mediated apoptosis. The research results will provide a theoretical basis for understanding the toxic effects and mechanisms of AgNPs on marine microalgae.

The impact of varying sizes of silver nanoparticles on the induction of cellular damage in Klebsiella pneumoniae involving diverse mechanisms

Abstract Silver nanoparticles (AgNPs) are extensively studied as potent antibacterial agents targeting antibiotic-resistant pathogens. Cellular damage induced through various mechanisms that can affect multiple cell components like the outer membrane, enzymes, and proteins is closely linked to their chemical and morphological characteristics. We investigated the impact of AgNPs’ size on their antibacterial effectiveness using two differently sized nanoparticles: silver nanoparticle-Citrus limon (AgCL) with an average size of 21 nm and silver nanoparticle-Citrus sinensis (AgCS) with an average size of 13 nm, derived from C. limon and C. sinensis through environmentally friendly methods. The study evaluated their antibacterial effects by assessing morphology changes via scanning electron microscopy, metabolic alterations using Fourier transform infrared (FT-IR) spectroscopy, and oxidative stress responses through biochemical markers in Klebsiella pneumoniae cells exposed to AgNPs. The results showed that both AgCL and AgCS exhibited remarkable antibacterial activity, evidenced by inhibition zones of 14 ± 1.5 and 16 ± 1.0 mm, respectively. Morphological changes in K. pneumoniae cells treated with AgNPs were size dependent, with notable alterations noted. FT-IR spectroscopy revealed size and concentration-dependent biochemical changes, particularly in shifts in functional groups involved in the fluidity of cell wall lipid, and protein structure. Exposure to AgNPs led to increased oxidative stress markers like lipid peroxides and reduced levels of enzymatic and non-enzymatic antioxidants, more prominently observed with smaller AgCS nanoparticles (13 nm). AgNPs induce oxidative stress and morphological changes in K. pneumoniae strains, with smaller nanoparticles demonstrating greater efficacy. These findings underscore the importance of nanoparticle size in optimizing the antibacterial properties against pathogens.

Method to Estimate Dislocation Densities from Images of α‐Ga2O3‐Based Corundum Oxides Using the Computer Vision YOLO Algorithm

This work applies the computer vision “You only look once” (YOLO) algorithm to extract bounding boxes around dislocations in weak‐beam dark‐field transmission electron microscopy (WBDF TEM) images of semiconductor thin films. A formula is derived to relate the sum of the relative heights of the bounding boxes to the dislocation densities in the films. WBDF TEM images reported in the literature and taken from our α‐Ga2O3 samples are divided into train, evaluation, and test datasets. Different models are trained using the train dataset and evaluated using the evaluation dataset to find the best confidence values, which are used to select the best model based on the performance against the test data set. For α‐Ga2O3 thin films, dislocation density output by this model is on average ≈58% of those estimated by the traditional Ham method. A factor of 4/π may contribute to the systematic underestimation of the model versus the Ham method.

Novel analysis tool for the distance of gold dimers controlled by the DNA strand length on the DNA origami.

Metallic nanoparticle dimers have been used to enhance the excitation rate of single-quantum emitters. The interparticle distance (d) of the dimers has a crucial influence on the signal enhancement. Therefore, precise control of d is desired for optimal performance. However, statistical analysis of d has been often restricted to a small number of dimers due to the lack of reliable automatic software tools. For this reason, we developed a novel analysis tool for automatic dimer analysis. Our approach combines particle detection by circle Hough transformation (CHT) with custom classification routines optimised for distinct types of particles. We applied our tool to scanning electron microscopy (SEM) images and achieved great agreement in dimer detection, reaching an agreement of around 97% between automatic analysis and manual inspection for more than 3000 metallic nanoparticle dimers on DNA origami controlled by a combination of multiple DNA strands. Our study revealed the effects of the strand length (L) on the distribution of d. Based on geometric consideration, we expected a strong correlation between L and the standard deviation (σ) of d. We could verify this correlation by characterising four dimer designs with different L while analysing more than 1000 dimers per specimen.

β-cryptoxanthin suppresses oxidative stress via activation of the Nrf2/HO-1 signaling pathway in diabetic kidney disease

ObjectivesThis study aims to explore the role and investigate mechanisms of β-Cryptoxanthin (BCX) in high glucose (HG)-induced podocyte injury and renal dysfunction.MethodsIn this study, db/db mice were orally treated with BCX. Blood glucose, body weight, urinary albumin creatinine ratio (ACR) were recorded to evaluate the mice renal function. The H&E, PAS staining, and transmission electron microscopy (TEM) were utilized to examine the effect of BCX on the morphological changes of glomeruli in db/db mice. In addition, reactive oxygen species (ROS) content, mitochondrial membrane potential (MMP) level, ATP level, and SA-β-gal staining were used to assess the podocyte oxidative damage, mitochondrial dysfunction and senescence. Furthermore, the effects of BCX on Nrf2/HO-1 signaling pathway were evaluated in vivo and in vitro through Western blotting, immunohistochemistry and immunofluorescence analysis.ResultsIn vivo, BCX reversed glomerular mesangial matrix expansion and reduced proteinuria in db/db mice, as well as decreased glomerular oxidative stress and kidney aging. Similarly, in vitro study showed that BCX effectively alleviated the oxidative stress, mitochondrial dysfunction, and senescence induced by HG in podocytes. Furthermore, we identified that the antioxidative effects of BCX are associated with the activation of Nrf2/HO-1 signaling pathway, and that Nrf2 knockdown partially abrogated the protective effects of BCX in vitro.ConclusionOur study demonstrated for the first time that BCX alleviates podocyte injury in DKD by promoting Nrf2/HO-1 signaling pathways. BCX may be a potential candidate compound for preventing Diabetic kidney disease (DKD).

CKS2 induces autophagy-mediated glutathione metabolic reprogramming to facilitate ferroptosis resistance in colon cancer.

Ferroptosis, a form of cell death characterized by lipid peroxidation, plays a crucial role in tumor suppression, offering novel avenues for cancer therapy. Previous studies have indicated that high levels of cyclin-dependent kinase subunit 2 (CKS2) promote the progression of various cancers. However, the potential interplay between CKS2 and ferroptosis in colon cancer (CC) remains unclear. Bioinformatics and RNA-seq analyses were employed to study genes associated with the ferroptosis signaling pathway. CKS2 expression was evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot (WB). The in vitro and in vivo effects of CKS2 on CC cells were assessed through the CCK-8 assay, colony formation assay, propidium iodide (PI) staining, BODIPY staining, DCFH-DA staining, and animal experiments. Additionally, the impact of CKS2 on autophagy and glutathione (GSH) metabolism was investigated using a transmission electron microscope (TEM), immunofluorescence (IF) assays, WB experiments, and relevant assay kits. CKS2 expression was elevated in CC, indicating a poor clinical outcome. Knockdown of CKS2 significantly enhanced Erastin-induced ferroptosis in CC cells, leading to reduced GSH metabolism. Conversely, CKS2 overexpression produced opposite effects. Mechanistically, CKS2-induced autophagy reinforced GSH metabolism, thereby increasing resistance to ferroptosis in CC cells. Furthermore, inhibiting CKS2 promoted tumor ferroptosis by downregulating GPX4 expression. Additionally, CKS2 knockdown effectively increased sorafenib-induced ferroptosis both in vitro and in vivo. CKS2 suppresses ferroptosis in CC by modulating GSH metabolism in both in vitro and in vivo settings. These findings offer new insights into targeting CKS2 for CC treatment and shed light on the mechanism of ferroptosis in CC.