Changes In Reactive Oxygen Species Research Articles

Exosomal miR-182-5p from breast cancer cells reprogram tumor-associated macrophages and promotes TNBC progression by targeting Notch1 in macrophages

IntroductionThe regulatory role of miR-182 in breast cancer malignancy and macrophage reprogramming is well-established. However, the mechanisms through which miR-182 overexpression in tumor cells influences macrophage polarization remain elusive.Material and methodsAfter transfection with miR-182-5p mimics, inhibitors, and controls for 24 hours, exosomes were extracted by differential centrifugation from transfected MDA-MB-231. Macrophages co-cultured with these exosomes to illustrate the regulative effects of exo-miR-182-5p reprogram macrophage. Furthermore, breast cancer cells co-cultured with exo-miR-182-5p reprogrammed M2 macrophages to demonstrate the effects of reprogrammed M2 macrophages to influence breast cancer progression. After all, these findings were validated in cell derived xenograft (CDX) BALB/C nude-mice model.ResultsIn this study, we demonstrate that exosome-derived miR-182-5p from TNBC cells reprograms M2 macrophage polarization through direct combined with Notch1, thereby enhancing breast cancer progression in vitro and in vivo. When co-cultured with exosomes from TNBC cells transfected with miR-182-5p mimics or inhibitors, macrophages showed altered Notch1/Hes1 pathway expression, leading to M2 polarization and subsequent changes in reactive oxygen species (ROS), inflammation, and other biochemical markers. Furthermore, breast cancer cells co-cultured with exosomes reprogrammed macrophages exhibited increased colony formation, migration, and invasion, as well as reduced apoptosis. These findings were validated in a BALB/C nude-mice model.ConclusionsThis study pioneers the elucidation of the feedback loop mechanism between breast cancer cells and macrophages mediated by the exosome-derived miR-182-5p/Notch1 pathway, highlighting its role in macrophage reprogramming. Although the therapeutic application of miR-182-5p inhibitors as anticancer agents remains in the early stages, targeting macrophage polarization represents a promising avenue for breast cancer therapy.

Antibacterial activity and mechanism of X33 antimicrobial oligopeptide against Acinetobacter baumannii

Acinetobacter baumannii is a pathogenic bacterium widespread in human environments, especially in intensive care units, and is associated with high morbidity and infection rates. Multiple drug resistance in A. baumannii frequently leads to the death of patients, making the development of multi-effect antibacterial agents against this bacterium a research hotspot. We have previously found that the X33 antimicrobial oligopeptide can effectively inhibit the growth of Penicillium digitatum and Candida albicans. Herein, we evaluated the antibacterial activity of X33 antimicrobial oligopeptide against A. baumannii by determining the minimum inhibitory concentration, inhibition zone, and growth curve. The increase in extracellular alkaline phosphatase and the leakage of intracellular compounds confirmed the effect of X33 antimicrobial oligopeptide on the cell wall and membrane. Changes in reactive oxygen species, malondialdehyde, ATP, reducing sugar, soluble protein, and pyruvate content demonstrated that the incubation with X33 antimicrobial oligopeptide affected energy metabolism and oxidative stress. Consistent with the physiological characteristics, transcriptomics analysis indicated that incubation with X33 antimicrobial oligopeptide significantly induced changes in the expression of 2339 genes, including 1262 upregulated and 1077 downregulated genes, which participate in oxidative phosphorylation, ribosome, quorum sensing, fatty acid degradation, glycolysis/gluconeogenesis, and citrate cycle pathways. These results provide a fundamental basis for investigating the mechanism of X33 antimicrobial oligopeptide as a potential drug against A. baumannii.

ENO2 promotes anoikis resistance in anaplastic thyroid cancer by maintaining redox homeostasis.

Anoikis presents a significant barrier in the metastasis of cancer. As the most aggressive type of thyroid cancer, anaplastic thyroid cancer (ATC) exhibits a high risk of metastasis and is characterized by high mortality. Therefore, investigating the molecular mechanisms of anoikis resistance in ATC is important for devising therapeutic targets in clinical research. Differentially Expressed Genes were screened in ATC cells under attached and detached culture conditions with RNA-seq. Investigate the impact of enolase 2 (ENO2) on apoptosis and spheroid formation by gain and loss of function. Changes of reactive oxygen species (ROS), glutathione (GSH) and nicotinamide adenine dinucleotide phosphate (NADPH) were detected to assess redox balance. The transcriptional regulatory role of signal transducer and activator of transcription 1 (STAT1) on ENO2 was validated through Dual-Luciferase Reporter Gene Assay. Explore the impact of ENO2 expression on the formation of lung metastases in nude mice. We found that the glycolysis process was activated in detached ATC cells. Several genes in the glycolysis process, particularly ENO2, a member of the enolase superfamily was upregulated in ATC cells cultured in suspension. The upregulation of ENO2 enabled the maintenance of redox balance by supplying GSH and NADPH, thereby preventing cells from undergoing anoikis. In terms of mechanism, the expression of STAT1 was enhanced in anoikis resistance cells, which in turn positively regulated the expression of ENO2. In vivo, ENO2-suppressed ATC cells resulted in a significantly lower rate of lung colonization compared to control ATC cells. Stable expression of ENO2 and the maintenance of redox balance played a pivotal role in facilitating anoikis resistance of ATC.

[Spermidine alleviates lipopolysaccharide-induced myocardial injury in mice by suppressing apoptosis, ROS production and ferroptosis].

To investigate the protective effect of spermidine against lipopolysaccharide (LPS)-induced myocardial injury in mice and the underlying mechanism. C57BL/6 mice subjected to intraperitoneal LPS injection with or without pretreatment with daily gavage of spermidine for 2 weeks were examined for myocardial pathologies using HE staining and transmission electron microscopy. In the cell experiment, cultured rat cardiomyocytes (H9c2 cells) were pretreated with 10 or 20 μmol/L spermidine before LPS exposure for 2 h, and the changes in cell viability and levels of lactate dehydrogenase (LDH) and cardiac troponin Ⅰ (cTNI) were assessed using CCK-8 kit, LDH detection kit and ELISA, respectively. Western blotting was performed to detect the changes in the expressions of Bax, Bcl-2, cleaved caspase-3, SLC7A11 and GPX4; the changes in reactive oxygen species (ROS) and Fe2+ levels were detected using fluorescent probes, and mitochondrial membrane potential of the cells was measured using JC-1 staining. Treatment of the mice with LPS induced obvious myocardial and mitochondrial damages, which were significantly alleviated by pretreatment with spermidine. In H9c2 cells, LPS exposure significantly lowered the cell viability, increased LDH and cTNI levels and expressions of Bax and cleaved caspase-3 levels, decreased expressions of Bcl-2, SLC7A11 and GPX4, increased ROS production and Fe2+ level (P < 0.05), and lowered mitochondrial membrane potential (all P < 0.05). These effects were significantly alleviated by SPD pretreatment of the cells prior to LPS exposure. Spermidine alleviates LPS-induced myocardial injury by suppressing cell apoptosis and inhibiting cellular ROS production and ferroptosis.

Physiological Responses Revealed Static Magnetic Fields Potentially Improving the Tolerance of Poplar Seedlings to Salt Stress

Magnetic fields play an important role in regulating plant growth and development, especially in improving plant stress tolerance. However, the physiological mechanism underlying the magnetic effects is still unclear. Here, we examined changes in reactive oxygen species (ROS) levels and ion flux in poplar (Populus × deltoides ‘Lulin-2’) seedling roots under salt stress in a static magnetic field (SMF). SMF treatment significantly increased seedling growth and mitigated the effects of salt stress on root growth. Furthermore, SMF treatment activated ROS and calcium signals in poplar roots. Relative to the SMF treatment group, control plants had significantly higher levels of cytoplasmic free Ca2+ ([Ca2+]cyt) and ROS following exposure to high salt concentrations. Under salt conditions, SMF treatment reduced increases in Na+ concentrations and maintained stable K+ and Ca2+ concentrations and K+/Na+ and Ca2+/Na+ ratios. NMT analysis suggests that SMF treatment may drive cation effluxes in poplar seedling roots. Susceptibility tests of Na+-transport inhibitors indicated that SMF treatment contributed to Na+ repulsion and H+ uptake under salt stress. Moreover, SMF exposure allowed roots to retain the ability to reduce salt-induced K+ and Ca2+ root effluxes, and qRT-PCR results demonstrate that SMF treatment can increase the expression of stress-responsive genes such as PtrRBOHF, PtrNHX1 and PtrHA5 in poplar seedlings. Therefore, we conclude that treating poplar seedlings with SMF can help them establish a stable tolerance to salt stress by regulating ROS, [Ca2+]cyt, and their regulatory networks. This study examined the physiological responses of poplar to SMF exposure under salt stress, providing insights into plant magnetobiological effects.

DriveLEDs: software for synchronous control and video acquisition of fluorescent signals

Current research uses various fluorescent biosensors to measure ion concentrations, neuronal activity, or cellular components in biological preparations. Several free solutions are available to researchers for recording or analysing fluorescent signals. However, when using different software packages, there are great difficulties in converting data between them. Problems also arise with linking and coordination of individual hardware devices into a single measurement system. Our paper presents useful software that allows to avoid most of these problems. It enables the recording, online visualization and preliminary analysis of fluorescent signals in brain cells and other experimental models. We describe and test software optimized for ratiometric measurements. The program selects device operating modes that allow reliable changes in illumination wavelength, camera image capture, and online graphical visualization of fluorescent signal amplitudes or ratiometric data. The performance of the software was successfully tested on mouse brain using two experimental models; i) simultaneous monitoring of intracellular chloride and hydrogen in transgenic mice expressing genetically encoded biosensor; ii) recording changes in reactive oxygen species during synaptic stimulation of neurons in mouse hippocampal slices. This software allows to overcome the incompatibility of the devices used and reduce the cost of experimental measurements. The software is completely original, easy to use and may be of interest to many scientists involved in the analysis of light-controlled signals in a variety of experimental models, including drug screening, epilepsy models, and other applications. The software is open-source product and can be obtained via GitHub: https://github.com/AndreyZakharovExp/DriveLEDs.

Adipose-Derived Mesenchymal Stem Cells Protect Endothelial Cells from Hypoxic Injury by Suppressing Terminal UPR In Vivo and In Vitro.

Adipose-derived stem cells (ASCs) have been used as a therapeutic intervention for peripheral artery disease (PAD) in clinical trials. To further explore the therapeutic mechanism of these mesenchymal multipotent stromal/stem cells in PAD, this study was designed to test the effect of xenogeneic ASCs extracted from human adipose tissue on hypoxic endothelial cells (ECs) and terminal unfolded protein response (UPR) in vitro and in an atherosclerosis-prone apolipoprotein E-deficient mice (ApoE-/- mice) hindlimb ischemia model in vivo. ASCs were added to Cobalt (II) chloride-treated ECs; then, metabolic activity, cell migration, and tube formation were evaluated. Fluorescence-based sensors were used to assess dynamic changes in Ca2+ levels in the cytosolic- and endoplasmic reticulum (ER) as well as changes in reactive oxygen species. Western blotting was used to observe the UPR pathway. To simulate an acute-on-chronic model of PAD, ApoE-/- mice were subjected to a double ligation of the femoral artery (DLFA). An assessment of functional recovery after DFLA was conducted, as well as histology of gastrocnemius. Hypoxia caused ER stress in ECs, but ASCs reduced it, thereby promoting cell survival. Treatment with ASCs ameliorated the effects of ischemia on muscle tissue in the ApoE-/- mice hindlimb ischemia model. Animals showed less muscle necrosis, less inflammation, and lower levels of muscle enzymes after ASC injection. In vitro and in vivo results revealed that all ER stress sensors (BIP, ATF6, CHOP, and XBP1) were activated. We also observed that the expression of these proteins was reduced in the ASCs treatment group. ASCs effectively alleviated endothelial dysfunction under hypoxic conditions by strengthening ATF6 and initiating a transcriptional program to restore ER homeostasis. In general, our data suggest that ASCs may be a meaningful treatment option for patients with PAD who do not have traditional revascularization options.

TRIM45 aggravates microglia pyroptosis via Atg5/NLRP3 axis in septic encephalopathy

BackgroundNeuroinflammation mediated by microglial pyroptosis is an important pathogenic mechanism of septic encephalopathy (SAE). It has been reported that TRIM45 is associated with tumours and inflammatory diseases. However, the role of TRIM45 in SAE and the relationship between TRIM45 and microglial pyroptosis are unknown. In this study, we found that TRIM45 played an important role in regulating microglial pyroptosis and the molecular mechanism.MethodsSAE was induced by intraperitoneal injection of LPS in WT and AAV-shTRIM45 mice. BV2 cells were treated with LPS/ATP in vitro. Cognitive function was assessed by the Morris water maze. Nissl staining was used to evaluate histological and structural lesions. ELISA was used to dectect neuroinflammation. qPCR was used to detect the mRNA levels of inflammatory cytokines, NLRP3, and autophagy genes. Western blotting and immunofluorescence analysis were used to analyse the expression of the proteins. Changes in reactive oxygen species (ROS) in cells were observed by flow cytometry. Changes in mitochondrial membrane potential in BV2 cells were detected by JC-1 staining. Peripheral blood mononuclear cells were extracted from blood by density gradient centrifugation and then used for qPCR, western blotting and flow detection. To further explore the mechanism, we used the overexpression plasmids TRIM45 and Atg5 as well as siRNA-TRIM45 and siRNA-Atg5 to analyse the downstream pathway of NLRP3. The protein and mRNA levels of TRIM45 in peripheral blood mononuclear cells from sepsis patients were examined.ResultsKnocking down TRIM45 protected against neuronal damage and cognitive impairment in septic mice. TRIM45 knockdown inhibited microglial pyroptosis and the secretion of inflammatory cytokines in vivo and in vitro, which was mediated by NLRP3/Gsdmd-N activation. Overexpression of TRIM45 could activate NLRP3 and downstream proteins. Further examination showed that TRIM45 regulated the activation of NLRP3 by altering Atg5 and regulating autophagic flux. It was also found that overexpression and knockdown of TRIM45 affected the changes in ROS and mitochondrial membrane potential. Thus, knocking down TRIM45 could reduce microglial pyroptosis, the secretion of proinflammatory cytokines, and neuronal damage and improve cognitive function. In addition, the level of TRIM45 protein in septic patients was increased. There was a positive linear correlation between APACHE II score and TRIM45, between SOFA score and TRIM45. Compared to group GCS > 9, level of TRIM45 were increased in group GCS ≤ 8.ConclusionTRIM45 plays a key role in neuroinflammation caused by LPS, and the mechanism may involve TRIM45-mediated exacerbation of microglial pyroptosis via the Atg5/NLRP3 axis.Graphical

The Armillaria response to Gastrodia elata is partially mediated by strigolactone-induced changes in reactive oxygen species

Armillaria root diseases, caused by Armillaria spp., pose a significant threat to woody plants worldwide and result in substantial economic losses. However, certain species in the genus Armillaria can establish a unique symbiotic relationship with Gastrodia elata, which is the only known example of a plant benefiting from Armillaria. Although various plant signals that play a role in this interaction have been identified, the mechanism remains largely unknown from the Armillaria’s perspective. In this study, we performed whole-genome sequencing of an Armillaria gallica strain named NRC001 isolated from G. elata. Comparative genomic analysis showed it is low-pathogenic Armillaria spp., which possesses 169 expanded gene families compared to high-pathogenic Armillaria spp. Among these expanded families, transcriptomic analysis revealed a significant increase in expression levels of four reactive oxygen species (ROS)-related gene families in A. gallica on G. elata compared to A. gallica on wood. Thus, a systematic survey of ROS-related gene families was carried out, and a total of 218 genes belonging to 44 ROS-related gene families in A. gallica were identified. Physiological experiments and transcriptome analysis showed that strigolactones (SLs) released by G. elata have a mediation impact on ROS, particularly enhancing the ROS scavenging activities by increasing the expression level and activity of several enzymes, such as catalase and glutathione reductase. Among the ROS-related genes, the aquaporin (AQP) is crucial as it is responsible for transporting hydrogen peroxide (H2O2) across the cell membrane. Five orthologs of AQP genes in A. gallica were identified and overexpressed in yeast. Only AgAQPA from the so-called ‘other aquaglyceroporin’ subfamily was demonstrated to be capable of mediating H2O2 transport in A. gallica. To our best knowledge, this is the first ‘other aquaglyceroporins’ gene in fungi to be identified as having transporter capacity. This study not only provides new insights into the mechanisms by which SL signaling regulates interactions between Armillaria and G. elata, but also sheds light on the function of fungal AQPs.

Porcine β-defensin-2 alleviates aflatoxin B1 induced intestinal mucosal damage via ROS-Erk1/2 signaling pathway

Aflatoxin B1 (AFB1) is a highly toxic fungal toxin that causes severe damage to animal intestines. Porcine beta-defensin-2 (pBD-2) is a well-studied antimicrobial peptide in pigs that can protect animal intestines and improve productivity. This study aimed to investigate the molecular mechanisms of pBD-2 in alleviating AFB1-induced oxidative stress and intestinal mucosal damage using porcine intestinal epithelial cells (IPEC-J2 cells) and Kunming (KM) mice. The maximum destructive concentration of AFB1 for IPEC-J2 cells and the optimal therapeutic concentration of pBD-2 were determined by CCK-8 and RT-qPCR. We then investigated the oxidative stress and intestinal damage induced by AFB1 and the alleviating effect of pBD-2 by detecting changes of reactive oxygen species (ROS), inflammatory cytokines, tight junction proteins (TJPs) and mucin. Finally, the molecular mechanism of pBD-2 mitigates AFB1-induced oxidative stress and intestinal mucosal damage were explored by adding ROS and Erk1/2 pathway inhibitors to comparative analysis. In vivo, the therapeutic effect of pBD-2 on AFB1-induced intestinal damage was analyzed from aspects such as average daily gain (ADG), pathological damage, inflammation, and mucosal barrier in KM mice. The study found that low doses of pBD-2 promoted cell proliferation and prevented AFB1-induced cell death, and pBD-2 significantly restored the feed conversion rate and ADG of KM mice reduced by long-term exposed AFB1. Increasing the intracellular ROS and the expression and phosphorylation of Erk1/2, AFB1 promoted inflammation by altering inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-8, and disrupted the mucosal barrier by interfering with Claudin-3, Occludin, and MUC2, while pBD-2 significantly reduced ROS and decreased the expression and phosphorylation of Erk1/2 to restored their expression to alleviate AFB1-induced oxidative stress and intestinal mucosal damage in IPEC-J2 cells and the small intestine of mice.

VDAC1 Protein Regulation of Oxidative Damage and Mitochondrial Dysfunction-Mediated Cytotoxicity by Silica Nanoparticles in SH-SY5Y Cells.

Silica nanoparticles (SiNPs) have been widely used in industry, electronics, and pharmaceutical industries. In addition, it is also widely used in medicine, tumor treatment and diagnosis, as well as other biomedical and biotechnology fields. The opportunities for people to contact SiNPs through iatrogenic, occupational, and environmental exposures are gradually increasing. The damage and biological effects of SiNPs on the nervous system have attracted widespread attention in the field of toxicology. Central nerve cells are rich in mitochondria. It is suggested that the effects of SiNPs on mitochondrial damage of nerve cells may involve the maintenance of neuronal membrane potential, the synthesis and operation of neurotransmitters, and the transmission of nerve pulses, and so on. We established an experimental model of SH-SY5Y cells to detect the cell survival rate, apoptosis, changes of reactive oxygen species and mitochondrial membrane potential, and the expression of mitochondrial function-related enzymes and proteins, so as to reveal the possible mechanism of SiNPs on neuronal mitochondrial damage. It was found that SiNPs could cause oxidative damage to cells and mitochondria, destroy some normal functions of mitochondria, and induce apoptosis in SH-SY5Y cells. The voltage-dependent anion channel 1(VDAC1) protein inhibitor DIDS could effectively reduce intracellular oxidative stress, such as the reduction of ROS content, and could also usefully restore some functional proteins of mitochondria to normal levels. The inhibition of VDAC1 protein may play an important role in the oxidative damage and dysfunction of neuronal mitochondria induced by SiNPs.

Ginsenoside Rg1 improved learning and memory ability and reduces neuronal apoptosis in epileptic rats through ERK/CREB/BDNF signal pathway

ObjectiveThis research was devoted to estimating the outcomes of ginsenoside Rg1 on learning and memory ability and neuronal apoptosis in epileptic rats through ERK/CREB/BDNF pathway. MethodsThe epileptic rats induced by lithium chloride were stochastically separated into model subgroup, ginsenoside Rg1 different dose subgroups. The ginsenoside Rg1 subgroups were given 20, 30 and 40 mg/kg ginsenoside Rg1 by gavage individually. Another 6 normal rats were selected as the control subgroup. The seizures of each subgroup were estimated. Morris water maze was utilized for estimating the changes of cognitive function changes of rats. The injury and apoptosis of hippocampal neurons in each subgroup were detected by Nissl and TUNEL assays. HE staining was applied for the structural and pathomorphological changes of hippocampal neurons detection. The oxidative stress level in hippocampus of rats was estimated by ELISA. DCFH-DA probe was applied for the changes of reactive oxygen species (ROS) in brain tissue detection. The Bcl-2, Bax, ERK, p-ERK, CREB, p-CREB and BDNF levels in cerebral cortex were estimated by western blot, and PD98059, a blocker of ERK pathway, was used to intervene. ResultsIn the control subgroup, Nissl bodies were abundant and evenly distributed, and cortical neurons were arranged neatly. In the model subgroup, the cytoplasmic staining of cortical neurons was insufficient and the arrangement of neurons was disordered. After treatment with ginsenoside Rg1, the morphology of neurons in the cerebral cortex was restored. The frequency of seizures, duration of seizures, Racine grade, escape latency, target quadrant residence time, MDA, TNF-α and ROS levels of cerebral cortex in the model subgroup boosted notablely versus the control subgroup. The frequency of crossing the original platform, the activity of SOD, the IL-10, p-ERK/ERK, p-CREB/CREB and BDNF levels in cerebral cortex were notablely lessened. The above-mentioned indexes in the ginsenoside Rg1 subgroup were notablely improved versus the model subgroup, and the three proteins levels in the PD98059 intervention subgroup were notablely lower. ConclusionGinsenoside Rg1 can improve cognitive dysfunction in epileptic rats, which may be concerned with ERK/CREB/BDNF pathway activation in cerebral cortex and lessening oxidative stress and inflammation.

Effects of photodynamic therapy using Red LED-light combined with hypocrellin B on apoptotic signaling in cutaneous squamous cell carcinoma A431 cells

BackgroundThe incidence of cutaneous squamous cell carcinoma (cSCC) has been demonstrating yearly increases. cSCC is a malignant cancer and exerts a major impact on patients' health and quality of life. Thus, the development and use of novel therapies in the treatment of cSCC are needed. It has been reported that LED photodynamic therapy (LED PDT) mediated by Hypocrellin B and its derivatives, a second-generation photosensitizer, can induce apoptosis in a variety of tumor cells, However, its potential pro-apoptotic effects on cSCC have yet to be investigated. ObjectiveThis study aims to investigate the pro-apoptotic effects and molecular mechanisms of HB-LED PDT in cutaneous squamous cell carcinoma A431 cells (Subsequent abbreviation A431 cells). Such information can provide an important theoretical foundation for the clinical translation of HB-LED PDT in the treatment of cSCC. Methods1. Effects of HB on A431 cells were determined using a Cell Counting Kit-8 assay, which method can indirectly reflect the number of living cells. In this way, this assay can then provide a means to identify the optimal concentrations of HB required for the induction of apoptosis in A431 cells. 2. The effects of HB-LED PDT on the morphology of A431 cells and changes in the nuclei after Hoechst33342 staining as determined using inverted fluorescent microscopy. 3. Use of the Annexin V-FITC test kit to detect levels of apoptosis in A431 cells in response to treatment with HB. Changes in reactive oxygen species and mitochondrial membrane potential following HB-LED PDT treatment in A431 cells were determined using fluorescence activated cell sorting (FACS). 4. Real-time quantitative PCR and Western Blot were applied to assess changes in several key factors involved in apoptosis including Bax, Bcl-2, and Caspase-3, at both transcription and translation levels. With these assays, it was possible to investigate the apoptotic signaling pathway in A431 cells in response to HB-LED PDT. ResultsHB-LED PDT inhibited proliferation activity and promoted nuclear fragmentation within these A431 cells. HB-LED PDT inhibited mitochondrial activity, increased reactive oxygen species production, and promoted apoptosis of A431 cells. In addition, several key factors in the apoptotic signaling pathway were increased at both the transcriptional and translational levels in A431 cells in response to the HB-LED PDT, indicating that the apoptotic signaling pathway was activated by HB-LED PDT. ConclusionHB-LED PDT induces apoptosis in A431 cells through a mitochondria-mediated apoptotic pathway. Such findings serve as an important foundation for the development of new approaches in the treatment of cSCC.

A prospective study of short-term apoplastic responses to ammonium treatment

The integration of external stimuli into plant cells has been extensively studied. Ammonium is a metabolic trigger because it affects plant nutrition status; on the contrary, it is also a stress factor inducing oxidative changes. Plants, upon quick reaction to the presence of ammonium, can avoid the development of toxicity symptoms, but their primary ammonium sensing mechanisms remain unknown. This study aimed to investigate the different signaling routes available in the extracellular space in response to supplying ammonium to plants.During short-term (30 min–24 h) ammonium treatment of Arabidopsis seedlings, no indication of oxidative stress development or cell wall modifications was observed. However, specific changes in reactive oxygen species (ROS) and redox status were observed in the apoplast, consequently leading to the activation of several ROS (RBOH, NQR), redox (MPK, OXI), and cell-wall (WAK, FER, THE, HERK) related genes. Therefore, it is expected that immediately after ammonium supply, a defense signaling route is initiated in the extracellular space. To conclude, the presence of ammonium is primarily perceived as a typical immune reaction.

Silver Nanomaterials Applied in the Field of Diagnosis and Treatment

Nanomaterials have great application prospect in biological medicine due to their strong permeability, repair and regeneration. In particular, nano silver has excellent antibacterial ability and no toxic effect, which makes it widely used. The specific content of this paper introduces a method of using silver nanoparticles to generate silver ions, and then make it react with hydrogen peroxide, through the colorimetric device to react with different color changes corresponding to different concentrations, through this biosensor to determine the concentration of such as cholesterol lactic acid and other substances.And when detecting miRNA,AuNPs were further modified by complementing the DNA (cDNA) of target mirnas. AgNPs could not be attracted to negatively charged DNA due to electrostatic repulsion by immobilized modified gold nanoparticles on the electrode surface. After analyzing the peak of silver stripping, miRNA can be accurately detected.Furthermore, chitosan coated silver nanoparticles were designed, and polyethylene glycol was used as a stabilizer. Finally, its good blood compatibility and antibacterial ability, as well as its cytotoxicity to cervical cancer cells were demonstrated by characterization.Finally, the accumulation and changes of reactive oxygen species (ROS) and reactive nitrogen (RNS) induced by silver nano are introduced, and it is confirmed by measurement that they are important factors in apoptosis of human pancreatic ductal adenocarcinoma.

Matrine alleviates oxidative stress and ferroptosis in severe acute pancreatitis-induced acute lung injury by activating the UCP2/SIRT3/PGC1α pathway.

Acute lung injury (ALI) is one of the most serious complications of severe acute pancreatitis (SAP). Matrine is well known for its powerful antioxidant and antiapoptotic properties, although its specific mechanism of action in SAP-ALI is unknown. In this study, we examined the effects of matrine on SAP-associated ALIand the specific signaling pathways implicated in SAP-induced ALI, such as oxidative stress, the UCP2-SIRT3-PGC1α pathway, and ferroptosis. The administration of caerulein and lipopolysaccharide (LPS) to UCP2-knockout (UCP2-/-) and wild-type (WT) mice that were pretreated with matrine resulted in pancreatic and lung injury. Changes in reactive oxygen species (ROS) levels, inflammation, and ferroptosis in BEAS-2B and MLE-12 cells were measured following knockdown or overexpression and LPS treatment. Matrine inhibited excessive ferroptosis and ROS production by activating the UCP2/SIRT3/PGC1α pathway while reducing histological damage, edema, myeloperoxidase activity and proinflammatory cytokine expression in the lung. UCP2 knockout decreased the anti-inflammatory properties of matrine and reduced the therapeutic effects of matrine on ROS accumulation and ferroptosis hyperactivation. LPS-induced ROS production and ferroptosis activation in BEAS-2B cells and MLE-12 cells were further enhanced by knockdown of UCP2, but this effect was rescued by UCP2 overexpression. This study demonstrated that matrine reduced inflammation, oxidative stress, and excessive ferroptosis in lung tissue during SAP by activating the UCP2/SIRT3/PGC1α pathway, demonstrating its therapeutic potential in SAP-ALI.

Integrated transcriptome and metabonomic analysis of key metabolic pathways in response to cadmium stress in novel buckwheat and cultivated species.

Buckwheat (Fagopyrum tataricum), an important food crop, also has medicinal uses. It is widely planted in Southwest China, overlapping with planting areas remarkably polluted by cadmium (Cd). Therefore, it is of great significance to study the response mechanism of buckwheat under Cd stress and further develop varieties with excellent Cd tolerance. In this study, two critical periods of Cd stress treatment (days 7 and 14 after Cd treatment) of cultivated buckwheat (Pinku-1, named K33) and perennial species (F. tatari-cymosum Q.F. Chen) (duoku, named DK19) were analyzed using transcriptome and metabolomics. The results showed that Cd stress led to changes in reactive oxygen species (ROS) and the chlorophyll system. Moreover, Cd-response genes related to stress response, amino acid metabolism, and ROS scavenging were enriched or activated in DK19. Transcriptome and metabolomic analyses highlighted the important role of galactose, lipid (glycerophosphatide metabolism and glycerophosphatide metabolism), and glutathione metabolism in response to Cd stress in buckwheat, which are significantly enriched at the gene and metabolic levels in DK19. The results of the present study provide valuable information for a better understanding of the molecular mechanisms underlying Cd tolerance in buckwheat and useful clues for the genetic improvement of drought tolerance in buckwheat.

Comparison of Immune Response of Litopenaeus vannamei Shrimp Naturally Infected with Vibrio Species, and after Being Fed with Florfenicol

The outbreaks caused by Vibrio spp. are a notable threat to the potential growth of the economy of penaeid culture, which is still controlled by the administration of antibiotics. At first, the infected group was subjected to phenotypic bacteriological examination with subsequent molecular identification via 16S rRNA gene sequencing, which confirmed four strains of Vibrio spp., V. atlanticus, V. natriegens, V. alginolyticus, and V. harveyi, from moribund-infected shrimp during mortality events in an Egyptian hatchery. To better understand the defense mechanism of the most effective antibiotic against Vibrio strains, the immune responses were compared and evaluated in infected Litopenaeus vannamei broodstock after being fed 5 mg kg−1 of florfenicol antibiotic, which was first determined through in vitro antibiogram tests. Therefore, our study aimed to determine the immune response of L. vannamei during Vibrio spp. infection in Egyptian hatcheries and after antibiotic medication. The parameters assessed were the total and differential hemocyte count (THC), granular cells (GC), semi-granular cells (SGC), and hyaline cells (HC). As well as the metabolic and immune enzymes: alanine aminotransferases (ALT), aspartate aminotransferases (AST), alkaline phosphatase (ALP), acid phosphatase (ACP), and lysozyme activity; an antioxidant index, such as superoxide dismutase (SOD) and glutathione (GSH); a phagocytic assay; changes in reactive oxygen species (ROS); and bactericidal activity in the hemolymph of the control, infected, and treated groups. Further evaluation of the mRNA expression levels of the prophenoloxidase (LvproPO), toll-like receptor 1 (LvToll1), and haemocyanin (LvHc) genes were performed in the hepatopancreas of the same groups. A significant drop in the THC, GC, SGC, and HC counts, as well as lysozyme and bactericidal activities, phagocytic assay, ROS, SOD, and GSH index, were represented in infected shrimp compared to control shrimp; however, a marked increase in the activity of ALT, AST, ALP, and ACP was observed. These activities were significantly restored in the treated shrimp compared to the infected shrimp. Nevertheless, no significant changes were noted in the transcriptional levels of the LvproPO and LvToll1 genes in the treated shrimp when compared to the infected shrimp; however, a significant suppression of the LvHc gene was noted. Our study aimed to determine the immune response of L. vannamei during Vibrio spp. infection in Egyptian hatcheries and after antibiotic medication. We concluded that florfenicol in medicated feed could be effective in controlling vibriosis and ameliorating the immune response of shrimp.

ABCB5+ mesenchymal stromal cells therapy protects from hypoxia by restoring Ca2+ homeostasis in vitro and in vivo

BackgroundHypoxia in ischemic disease impairs Ca2+ homeostasis and may promote angiogenesis. The therapeutic efficacy of mesenchymal stromal cells (MSCs) in peripheral arterial occlusive disease is well established, yet its influence on cellular Ca2+ homeostasis remains to be elucidated. We addressed the influence of ATP-binding cassette subfamily B member 5 positive mesenchymal stromal cells (ABCB5+ MSCs) on Ca2+ homeostasis in hypoxic human umbilical vein endothelial cells (HUVECs) in vitro and in vivo.MethodsHypoxia was induced in HUVECs by Cobalt (II) chloride (CoCl2) or Deferoxamine (DFO). Dynamic changes in the cytosolic- and endoplasmic reticulum (ER) Ca2+ and changes in reactive oxygen species were assessed by appropriate fluorescence-based sensors. Metabolic activity, cell migration, and tube formation were assessed by standard assays. Acute-on-chronic ischemia in Apolipoprotein E knock-out (ApoE−/−) mice was performed by double ligation of the right femoral artery (DFLA). ABCB5+ MSC cells were injected into the ischemic limb. Functional recovery after DFLA and histology of gastrocnemius and aorta were assessed.ResultsHypoxia-induced impairment of cytosolic and ER Ca2+ were restored by ABCB5+ MSCs or their conditioned medium. Similar was found for changes in intracellular ROS production, metabolic activity, migratory ability and tube formation. The restoration was paralleled by an increased expression of the Ca2+ transporter Sarco-/endoplasmic reticulum ATPase 2a (SERCA2a) and the phosphorylation of Phospholamban (PLN). In acute-on-chronic ischemia, ABCB5+ MSCs treated mice showed a higher microvascular density, increased SERCA2a expression and PLN phosphorylation relative to untreated controls.ConclusionsABCB5+ MSCs therapy can restore cellular Ca2+ homeostasis, which may beneficially affect the angiogenic function of endothelial cells under hypoxia in vitro and in vivo.

NSUN2 alleviates doxorubicin-induced myocardial injury through Nrf2-mediated antioxidant stress

Doxorubicin (DOX) is a commonly used antitumor drug, but its application has been limited because of its strong cardiac damage. This study aims to explore the role of NSUN2 in DOX-induced heart injury. C57BL/6J mice were intraperitoneally injected with 20 mg/Kg DOX to induce heart injury. After 3 days, the cardiac function, cardiac histopathology, myocardial apoptosis, and the expression level of NSUN2 were detected. In vitro, H9C2 cells were transfected with NSUN2 siRNA or overexpressed lentivirus and then treated with 500 ng/ml DOX. After 24 h, the changes in reactive oxygen species (ROS), apoptosis, and NSUN2 expression were detected. After DOX treatment, both in vitro and in vivo experiments showed that the cardiac function decreased, the number of apoptotic cells increased, and the expression level of NSUN2 increased. Interfering the expression of NSUN2 by siRNA promoted DOX-induced heart injury, while overexpression of NSUN2 could inhibit DOX-induced heart injury. Further study showed that NSUN2 promoted antioxidative stress by upregulating the Nrf2 protein level. In addition, NSUN2 overexpression could increase the half-life of Nrf2 mRNA. m5C RNA methylation immunoprecipitation (MeRIP) also showed that the level of Nrf2 m5C mRNA was significantly increased in NSUN2 overexpressed group when compared to the GFP group. NSUN2 enhances the expression of Nrf2 by promoting Nrf2 mRNA m5C modification and enhances its antioxidative stress effect to alleviate DOX-induced myocardial injury.