ABSTRACT
With the rapid global spread of COVID-19 and the continuous emergence of variants, there is an urgent need to develop safe and effective vaccines. Here, we developed a novel mRNA vaccine, HC009, based on new formulation by the QTsome delivery platform. Immunogenicity results showed that the prime-boost immunization strategy with HC009 was able to induce robust and durable humoral immunity, as well as Th1-biased cellular responses in rodents or non-human primates (NHPs). After further challenge with live SARS-CoV-2 virus, HC009 provided adequate protection against virus infection in hACE2 transgenic mice. Therefore, HC009 could provide significant immune protection against SARS-CoV-2.
Subject(s)
Antibodies, Viral , COVID-19 Vaccines , COVID-19 , Immunogenicity, Vaccine , Mice, Transgenic , SARS-CoV-2 , mRNA Vaccines , Animals , SARS-CoV-2/immunology , COVID-19 Vaccines/immunology , COVID-19/prevention & control , COVID-19/immunology , Mice , mRNA Vaccines/immunology , Antibodies, Viral/immunology , Antibodies, Viral/blood , Humans , Vaccines, Synthetic/immunology , Vaccines, Synthetic/administration & dosage , Immunity, Humoral , Female , Antibodies, Neutralizing/immunology , Antibodies, Neutralizing/blood , Mice, Inbred BALB C , Vaccine EfficacyABSTRACT
Oxidative stress (OS) is a major mediator of secondary brain injury following intracerebral hemorrhage (ICH). Thus, antioxidant therapy is emerging as an attractive strategy to combat ICH. To achieve both reactive oxygen species (ROS) scavenging ability and on-demand drug release ability, we constructed a novel polydopamine (PDA)-coated diselenide-bridged mesoporous silica nanoparticle (DSeMSN) drug delivery system (PDA-DSeMSN). Edaravone (Eda) was blocked in the pores of DSeMSN by covering the pores with PDA as a gatekeeper. The drug maintained nearly "zero release" before reaching the lesion site, while in the ROS-enriched circumstances, the PDA shell went through degradation and the doped diselenide bonds broke up, triggering the disintegration of nanoparticles and leading to Eda release. Interestingly, the ROS-degradable property of the PDA shell and diselenide bond endowed the system with enhanced ROS-eliminating capacity. The synergistic effect of ROS-responsive drug delivery and ROS-scavenging PDA-DSeMSN showed efficient antioxidative and mitochondria protective performance without apparent toxicity in vitro. Importantly, PDA-DSeMSN@Eda through intravenous administration specifically accumulated in perihematomal sites and demonstrated robust neuroprotection in an ICH mouse model through antioxidative and antiapoptotic effects with high biological safety. Thus, the PDA-DSeMSN platform holds tremendous potential as an excellent carrier for on-demand delivery of drugs and provides a new and effective strategy for the clinical treatment of ICH.
Subject(s)
Cerebral Hemorrhage , Edaravone , Indoles , Nanoparticles , Reactive Oxygen Species , Silicon Dioxide , Animals , Silicon Dioxide/chemistry , Cerebral Hemorrhage/drug therapy , Cerebral Hemorrhage/pathology , Reactive Oxygen Species/metabolism , Mice , Nanoparticles/chemistry , Edaravone/chemistry , Edaravone/pharmacology , Indoles/chemistry , Indoles/pharmacology , Porosity , Polymers/chemistry , Polymers/pharmacology , Free Radical Scavengers/chemistry , Free Radical Scavengers/pharmacology , Male , Antioxidants/chemistry , Antioxidants/pharmacology , Oxidative Stress/drug effectsABSTRACT
BACKGROUND: Evidence indicates that the risk of developing a secondary ovarian cancer (OC) is correlated with estrogen receptor (ER) status. However, the clinical significance of the relationship between ER-associated breast cancer (BC) and clear cell ovarian cancer (CCOC) remains elusive. METHODS: Independent single nucleotide polymorphisms (SNPs) strongly correlated with exposure were extracted, and those associated with confounders and outcomes were removed using the PhenoScanner database. SNP effects were extracted from the outcome datasets with minor allele frequency > 0.01 as the filtration criterion. Next, valid instrumental variables (IVs) were obtained by harmonizing exposure and outcome effects and further filtered based on F-statistics (> 10). Mendelian randomization (MR) assessment of valid IVs was carried out using inverse variance weighted (IVW), MR Egger (ME), weighted median (WM), and multiplicative random effects-inverse variance weighted (MRE-IVW) methods. For sensitivity analysis and visualization of MR findings, a heterogeneity test, a pleiotropy test, a leave-one-out test, scatter plots, forest plots, and funnel plots were employed. RESULTS: MR analyses with all four methods revealed that CCOC was not causally associated with ER-negative BC (IVW results: odds ratio (OR) = 0.89, 95% confidence interval (CI) = 0.66-1.20, P = 0.431) or ER-positive BC (IVW results: OR = 0.99, 95% CI = 0.88-1.12, P = 0.901). F-statistics were computed for each valid IV, all of which exceeded 10. The stability and reliability of the results were confirmed by sensitivity analysis. CONCLUSIONS: Our findings indicated that CCOC dids not have a causal association with ER-associated BC. The absence of a definitive causal link between ER-associated BC and CCOC suggested a minimal true causal influence of ER-associated BC exposure factors on CCOC. These results indicated that individuals afflicted by ER-associated BC could alleviate concerns regarding the developing of CCOC, thereby aiding in preserving their mental well-being stability and optimizing the efficacy of primary disease treatment.
ABSTRACT
Underground coal seam mining significantly alters the stress and energy distribution within the overlying rock, leading to eventual structural degradation. Therefore, it is imperative to quantitatively identify the temporal and spatial characteristics of stress evolution of overlying rock caused by mining. This paper introduces a novel rock stress model integrating entropy and a spatial-temporal cube. Similar material model tests are used to identify the abrupt entropy changes within the mining rock, and the trend analysis is carried out to describe the spatial-temporal evolution law of stress during mining. Experimental findings indicate elevated stress levels in the unmined rock preceding and following the panel, as well as within specific rock strata above it. Definitively, dynamic stress arches within the surrounding rock of the stope predominantly bear and distribute the load and pressure from the overlying rock, and each stress mutation is accompanied by a sudden stress entropy change. Over time, z-score shows that the noticeable reduction in mining-induced overburden stress becomes increasingly pronounced, especially in the water-conducting fracture zone. The model's bifurcation set serves as the comprehensive criterion for the entropy-induced sudden changes in the rock system, signifying overall failure.
ABSTRACT
Composite polymer solid electrolytes (CPEs), possessing good rigid flexible, are expected to be used in solid-state lithium-metal batteries. The integration of fillers into polymer matrices emerges as a dominant strategy to improve Li+ transport and form a Li+-conducting electrode-electrolyte interface. However, challenges arise as traditional fillers: 1) inorganic fillers, characterized by high interfacial energy, induce agglomeration; 2) organic fillers, with elevated crystallinity, impede intrinsic ionic conductivity, both severely hindering Li+ migration. Here, a concept of super-ionic conductor soft filler, utilizing a Li+ conductivity nanocellulose (Li-NC) as a model, is introduced which exhibits super-ionic conductivity. Li-NC anchors anions, and enhances Li+ transport speed, and assists in the integration of cathode-electrolyte electrodes for room temperature solid-state batteries. The tough dual-channel Li+ transport electrolyte (TDCT) with Li-NC and polyvinylidene fluoride (PVDF) demonstrates a high Li+ transfer number (0.79) due to the synergistic coordination mechanism in Li+ transport. Integrated electrodes' design enables stable performance in LiNi0.5Co0.2Mn0.3O2|Li cells, with 720 cycles at 0.5 C, and 88.8% capacity retention. Furthermore, the lifespan of Li|TDCT|Li cells over 4000 h and Li-rich Li1.2Ni0.13Co0.13Mn0.54O2|Li cells exhibits excellent performance, proving the practical application potential of soft filler for high energy density solid-state lithium-metal batteries at room temperature.
ABSTRACT
The unstable interface between Li metal and ethylene carbonate (EC)-based electrolytes triggers continuous side reactions and uncontrolled dendrite growth, significantly impacting the lifespan of Li metal batteries (LMBs). Herein, a bipolar polymeric protective layer (BPPL) is developed using cyanoethyl (-CH2CH2C≡N) and hydroxyl (-OH) polar groups, aiming to prevent EC-induced corrosion and facilitating rapid, uniform Li+ ion transport. Hydrogen-bonding interactions between -OH and EC facilitates the Li+ desolvation process and effectively traps free EC molecules, thereby eliminating parasitic reactions. Meanwhile, the -CH2CH2C≡N group anchors TFSI- anions through ion-dipole interactions, enhancing Li+ transport and eliminating concentration polarization, ultimately suppressing the growth of Li dendrite. This BPPL enabling Li|Li cell stable cycling over 750 cycles at 10â mA cm-2 for 2â mAh cm-2. The Li|LiNi0.8Mn0.1Co0.1O2 and Li|LiFePO4 full cells display superior electrochemical performance. The BPPL provides a practical strategy to enhanced stability and performance in LMBs application.
ABSTRACT
BACKGROUND: Systemic lupus erythematosus is an immunologically dysregulated disease characterized by the presence of multiple autoantibodies. In SLE, B lymphocytes contribute to the dysregulated production of autoantibodies and cytokines. Recently, we discovered that miR-99a-3p binds to both EIF4EBP1 and NCAPG mRNA and that lowering miR-99a-3p can promote B cell autophagy in SLE by increasing EIF4EBP1 expression. However, the functions of miR-99a-3p and NCAPG in SLE have not been extensively investigated. OBJECTIVE: This work aims to evaluate the levels of miR-99a-3p and NCAPG expression in SLE B cells and to determine whether the aberrant expression of miR-99a-3p and NCAPG contributes to the pathological mechanisms in SLE. METHODS: B lymphocytes were obtained through immunomagnetic negative selection. Using RT-qPCR, miR-99a-3p and NCAPG mRNA expressions in B lymphocytes and in the BALL-1 cell line were measured. To determine the relative abundance of NCAPG, PI3K, p-PI3K, AKT, and p-AKT, we normalize them to the level of ß-actin using Western blotting. Evaluation of miR-99a-3p and NCAPG's impact on cell proliferation was done utilizing CCK-8 assay. Using flow cytometry, the cell cycle and apoptosis were both measured. RESULTS: Comparing SLE B cells to healthy controls, miR-99a-3p expression was significantly downregulated. Additionally, it was observed that SLE B cells had significantly higher NCAPG mRNA expression. Blocking miR-99a-3p expression in BALL-1 cells with an antagomir elevated NCAPG expression, facilitated PI3K/AKT pathway activation, improved cell proliferation, raised the fraction of S-phase cells, and prevented cell apoptosis. The opposite effects of upregulated miR-99a-3p levels on BALL-1 cells were observed by using an agomir. Furthermore, the effect of decreased miR-99a-3p expression on cell proliferation was partially mediated by elevating NCAPG levels and activating the PI3K/AKT pathway. CONCLUSION: Our research indicates that lower miR-99a-3p expression in SLE B cells appears to boost B cell number via the NCAPG and PI3K/AKT pathways.
Subject(s)
Lupus Erythematosus, Systemic , MicroRNAs , Humans , Autoantibodies/pharmacology , Cell Cycle Proteins/metabolism , Cell Cycle Proteins/pharmacology , Cell Proliferation/genetics , MicroRNAs/genetics , MicroRNAs/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/genetics , RNA, Messenger , Signal TransductionABSTRACT
BACKGROUND: Blood brain barrier (BBB) breakdown is one of the key mechanisms of secondary brain injury following intracerebral hemorrhage (ICH). Astrocytes interact with endothelial and regulate BBB integrity via paracrine signaling factors. More and more studies reveal astrocyte-derived extracellular vesicles (ADEVs) as an important way of intercellular communication. However, the role of ADEV in BBB integrity after ICH remains unclear. METHODS: ADEVs were obtained from astrocytes with or without oxygen and glucose deprivation (OGD) pre-stimulation and the role of ADEVs in ICH was investigated using ICH mice model and ICH cell model. The potential regulatory effect of ADEVs on endothelial barrier integrity was identified by TEER, western blot and immunofluorescence in vitro. In vivo, functional evaluation, Evans-blue leakage and tight junction proteins (TJPs) expression were analyzed. MiRNA sequencing revealed that microRNA-27a-3p (miR-27a-3p) was differentially expressed miRNA in the EVs from OGD-pretreated astrocytes compared with normal control. The regulatory mechanism of miR-27a-3p was assessed using Luciferase assay, RT-PCR, western blot and immunofluorescence. RESULTS: OGD-activated astrocytes reduced hemin-induced endothelial hyper-permeability through secreting EVs. OGD-activated ADEVs alleviated BBB dysfunction after ICH in vivo and in vitro. MicroRNA microarray analysis indicated that miR-27a-3p is a major component that was highly expressed miRNA in OGD pretreated-ADEVs. OGD-ADEVs mitigated BBB injury through transferring miR-27a-3p into bEnd.3 cells and regulating ARHGAP25/Wnt/ß-catenin pathway. CONCLUSION: Taken together, these findings firstly revealed that miR-27a-3p, as one of the main components of OGD-pretreated ADEVs, attenuated BBB destruction and improved neurological deficits following ICH by regulating endothelial ARHGAP25/Wnt/ß-catenin axis. OGD-ADEVs might be a novel strategy for the treatment of ICH. this study implicates that EVs from OGD pre-stimulated astrocytes.
Subject(s)
Exosomes , MicroRNAs , Animals , Mice , Blood-Brain Barrier/metabolism , Astrocytes/metabolism , beta Catenin/metabolism , Endothelial Cells/metabolism , Exosomes/metabolism , Oxygen/metabolism , Glucose , MicroRNAs/genetics , MicroRNAs/metabolism , MicroRNAs/pharmacology , Cerebral Hemorrhage/metabolismABSTRACT
Titanium (Ti) implants have been extensively used after surgical operations. Its surface bioactivity is of importance to facilitate integration with surrounding bone tissue, and ultimately ensure stability and long-term functionality of the implant. The plasmid DNA-activated matrix (DAM) coating on the surface could benefit osseointegration but is still trapped by poor transfection for further application, especially on the bone marrow mesenchymal stem cells (BMSCs) in vivo practical conditions. Herein, we constructed a DAM on the surface of fibrous-grained titanium (FG Ti) composed of phase-transition lysozyme (P) as adhesive, cationic arginine-rich lipid (RLS) as the transfection agent and plasmid DNA (pDNA) for bone morphology protein 2 (BMP2) expression. The cationic lipid RLS improved up to 30-fold higher transfection than that of commercial reagents (Lipofectamine 2000 and polyethyleneimine) on MSC. And importantly, Ti surface topology not only promotes the DAM to achieve high transfection efficiency (â¼75.7% positive cells) on MSC due to the favorable combination but also reserves its contact induction effect for osteoblasts. Upon further exploration, the fibrous topology on FG Ti could boost pDNA uptake for gene transfection, and cell migration in MSC through cytoskeleton remodeling and induce contact guidance for enhanced osteointegration. At the same time, the cationic RLS together with adhesive P were both antibacterial, showing up to 90% inhibition rate against Escherichia coli and Staphylococcus aureus with reduced adherent microorganisms and disrupted bacteria. Finally, the FG Ti-P/pBMP2 implant achieved accelerated bone healing capacities through highly efficient gene delivery, aligned surface topological structure and increased antimicrobial properties in a rat femoral condylar defect model.
ABSTRACT
In the pursuit of advancing materials for methane storage, a critical consideration arises given the prominence of natural gas (NG) as a clean transportation fuel, which holds substantial potential for alleviating the strain on both energy resources and the environment in the forthcoming decade. In this context, a novel approach is undertaken, employing the rigid triptycene as a foundational building block. This strategy is coupled with the incorporation of dichloromethane and 1,3-dichloropropane, serving as rigid and flexible linkers, respectively. This combination not only enables cost-effective fabrication but also expedites the creation of two distinct triptycene-based hypercrosslinked polymers (HCPs), identified as PTN-70 and PTN-71. Surprisingly, despite PTN-71 manifesting an inferior Brunauer-Emmett-Teller (BET) surface area when compared to the rigidly linked PTN-70, it showcases remarkably enhanced methane adsorption capabilities, particularly under high-pressure conditions. At a temperature of 275 K and a pressure of 95 bars, PTN-71 demonstrates an impressive methane adsorption capacity of 329 cm3 g-1. This exceptional performance is attributed to the unique flexible network structure of PTN-71, which exhibits a pronounced swelling response when subjected to elevated pressure conditions, thus elucidating its superior methane adsorption characteristics. The development of these advanced materials not only signifies a significant stride in the realm of methane storage but also underscores the importance of tailoring the structural attributes of hypercrosslinked polymers for optimized gas adsorption performance.
ABSTRACT
BACKGROUND: There are no standards for evaluating skin photoaging. Dermoscopy is a non-invasive detection method that might be useful for evaluating photoaging. OBJECTIVE: To assess the correlation between the dermoscopic evaluation of photoaging and clinical and pathological evaluations. METHODS: The age, clinical evaluation (Fitzpatrick classification, Glogau Photoaging Classification, and Chung's standardized image ruler), histopathology (Masson staining and MMP-1 immunohistochemistry), and dermoscopy (Hu's and Isik's) of 40 donor skin samples were analyzed statistically, and Spearman rank correlation analysis was performed. RESULTS: There was a robust correlation between the total Hu scores and Isik dermoscopy. The correlation of dermoscopy with histopathology was higher than that of clinical evaluation methods. There is a strong correlation between telangiectases and lentigo. Xerosis, superficial wrinkle, diffuse erythema, telangiectases, and reticular pigmentation were significantly correlated with the three clinical evaluation methods. Superficial wrinkles were correlated with Masson, MMP-1, various clinical indicators, and other dermoscopic items. CONCLUSION: There is a good correlation between dermoscopy and clinical and histopathological examination. Dermoscopy might help evaluate skin photoaging.
Subject(s)
Lentigo , Skin Aging , Skin Neoplasms , Telangiectasis , Humans , Matrix Metalloproteinase 1 , Dermoscopy/methods , Telangiectasis/diagnostic imaging , Skin Neoplasms/pathologyABSTRACT
Garnet solid electrolyte Li6.4 La3 Zr1.4 Ta0.6 O12 (LLZTO) is an excellent inorganic ceramic-type solid electrolyte; however, the presence of Li2 CO3 impurities on its surface hinders Li-ion transport and increases the interface impedance. In contrast to traditional methods of mechanical polishing, acid corrosion, and high-temperature reduction for removing Li2 CO3 , herein, a straightforward "waste-to-treasure" strategy is proposed to transform Li2 CO3 into Li3 PO4 and LiF in LiPF6 solution under 60 °C. It is found that the formation of Li3 PO4 during LLZTO pretreatment facilitates rapid Li-ion transport and enhances ionic conductivity, and the LLZTO/PAN composite polymer electrolyte shows the highest Li-ion transference number of 0.63. Additionally, the dense LiF layer serves to safeguard the internal garnet solid electrolyte against solvent decomposition-induced chemical adsorption. Symmetric Li/Li cells assembled with treated LLZTO/PAN composite electrolyte exhibit a critical current density of 1.1 mA cm-2 and a long lifespan of up to 700 h at a current density of 0.2 mA cm-2 . The Li/LiFePO4 solid-state cells demonstrate stable cycling performances for 141 mAh g-1 at 0.5 C, with capacity retention of 93.6% after 190 cycles. This work presents a novel approach to converting waste into valuable resources, offering the advantages of simple processes, and minimal side reactions.
ABSTRACT
Systemic Lupus Erythematosus (SLE) is an autoimmune sickness with unclear pathogenesis. The goal of this research was to reveal the heterogeneity of immune cells in SLE patients of Han and Zang nationality by single-cell RNA sequencing (scRNA-seq) and bioinformatics profiling. METHODS: A total of 94,102 peripheral blood mononuclear cells (PBMCs) from six volunteers with SLE (3 Zang, 3 Han) and six healthy controls were first conducted through scRNA-seq analysis. The immune cell subsets in the pathogenesis of SLE were analyzed as well. Real-time quantitative PCR (RT-qPCR) was applied to confirm the results of sc-RNA seq analysis. RESULTS: For the Tibetan samples, the ratios of Naïve CD4 RPS4Y1 cells, Naïve CD4 cells, Memory BC CD24 and Memory BC differed significantly between the SLE and control samples, while that of CD8 CTL MAL cells was significantly different between the two groups in Han nationality samples. Variable differentiation states of CD8 CTL MAL cells, CD8 CTL GZMK cells, and Naïve CD4 cells were detected through pseudotime analysis. Moreover, T-cell receptor (TCR) abundance was notably higher in Tibetan SLE specimens than that in controls, while B-cell receptor (BCR) abundance in Tibetan and Han samples was higher than in control groups. CONCLUSIONS: In summary, the immune cellular heterogeneity of SLE patients both Han and Zang nationality was explored based on various bioinformatics approaches, providing new perspectives for immunological characteristics of SLE among different ethnic groups.
Subject(s)
Leukocytes, Mononuclear , Lupus Erythematosus, Systemic , Humans , Cell Differentiation , Ethnicity , Lupus Erythematosus, Systemic/genetics , Sequence Analysis, RNAABSTRACT
The serine/threonine kinase Akt is an important component of the insulin signalling pathway (ISP) in regulating insect metabolism, growth, and reproduction. The psocid Liposcelis entomophila (Enderlein) is a distasteful stored products pest for its fecundity. However, the molecular mechanism of Akt that controls vitellogenesis and oviposition in L. entomophila remains obscure. In this study, the function of the Akt gene in the female reproduction of L. entomophila (designated as LeAkt) was characterized and investigated. LeAkt contains a 1587 bp open reading frame encoding a 529 amino acid protein that possesses a conserved Pleckstrin Homology domain (PH) and a Ser/Thr-type protein kinase (S_TKc) domain. The mRNA expression of LeAkt was the highest in female adult stages and peaked for 7-day female adults. In female adult tissues, LeAkt was highly expressed in the head and the ovary, indicating that LeAkt was closely correlated with female ovarian development. LeAkt transcription level was significantly suppressed by oral feeding on artificial diets mixed with dsRNA-LeAkt. RNAi-mediated silencing of LeAkt led to a severe inhibition of vitellogenein (Vg) expression and ovarian development, together with lower fecundity and hatchability compared to that of the normal feeding group, suggesting a critical role for LeAkt in L. entomophila reproduction. Further studies revealed that LeAkt silencing significantly decreased the mRNA levels of several signalling and biosynthetic genes in the juvenile hormone (JH) signalling pathway, such as methoprene-tolerant (LeMet), krüppel homolog 1 (LeKr-h1) and JH methyltransferase (LeJHAMT), leading to a severe inhibition of JH biosynthesis in L. entomophila female adults. These results suggested that LeAkt was affecting JH synthesis, thereby influencing Vg synthesis and ultimately L. entomophila reproduction.
Subject(s)
Juvenile Hormones , Proto-Oncogene Proteins c-akt , Animals , Proto-Oncogene Proteins c-akt/genetics , Juvenile Hormones/metabolism , Fertility , RNA, Messenger , Serine , Insect Proteins/genetics , Insect Proteins/metabolismABSTRACT
Organoids, which are clumps of cells formed after in vitro 3D culture utilizing autologous tissue and stem cells, possess the 3D structure and corresponding functional and genetic features of the original tissue and organ. This model has immense potential in modeling the ontogeny of specific organismal organs, as well as in drug screening and studying molecular mechanisms. The newly developed concept of bone organoids, a special type of complex hard tissues that can be created in vitro using tissue engineering 3D culture technology, mimics the complex biological functions of bone tissue in vivo. These bone organoids are highly useful in elucidating the regulatory mechanisms of bone regeneration, screening tissue engineering materials, and promoting bone regeneration and repair. They offer promising applications in bone regeneration research.
Subject(s)
Organoids , Tissue Engineering , Stem Cells , EngineeringABSTRACT
A new sensor has been developed to detect miRNA-15 using nanoelectrodes and a hairpin DNA-based electrochemical amplification technique. By utilizing a complex DNA cylinder connected with hairpin DNA1, the sensor is able to absorb more methylene blue (MB) than simple double-stranded DNA. Another hairpin DNA2 is modified on an Au nanoelectrode surface and, when miRNA-15 is introduced, it triggers a chain reaction. This reaction unlocks two hairpins alternatively to polymerize into a complex structure that attaches more MB. The miRNA-15 is then replaced by DNA1 due to strand displacement reactions and continues to react with the next DNA2 to achieve circular amplification. The electrochemical signal from MB oxidation has a linear relationship with the miRNA-15 concentrations, making it possible to detect miRNA-15. Moreover, this method can be readily adapted for the detection of various other miRNA species. The newly devised nanosensor holds promising applications for the in vivo detection of miRNA-15 within biological systems, which is achieved by leveraging the advantageous characteristics of nanoelectrodes, including their low resistance-capacitance time constant, rapid mass transfer kinetics, and small diameter.
Subject(s)
Biosensing Techniques , MicroRNAs , MicroRNAs/genetics , MicroRNAs/chemistry , Gold/chemistry , Electrochemical Techniques/methods , DNA/genetics , DNA/chemistry , Methylene Blue/chemistry , Biosensing Techniques/methods , Limit of DetectionABSTRACT
Photoaging is unique to the skin and is accompanied by an increased risk of tumors. To explore the transcriptomic regulatory mechanism of skin photoaging, the epidermis, and dermis of 16 healthy donors (eight exposed and eight non-exposed) were surgically excised and detected using total RNA-Seq. Weighted gene co-expression network analysis (WGCNA) identified the most relevant modules with exposure. The hub genes were identified using correlation, p-value, and enrichment analysis. The critical genes were identified using Support Vector Machine-Recursive Feature Elimination (SVM-RFE) and least absolute shrinkage and selection operator (LASSO) regression, then enriched using single-gene GSEA. A competitive endogenous RNA (ceRNA) network was constructed and validated using qRT-PCR. Compared with non-exposed sites, 430 mRNAs, 168 lncRNAs, and 136 miRNAs were differentially expressed in the exposed skin. WGCNA identified the module MEthistle and 12 intersecting genes from the 71 genes in this module. The enriched pathways were related to muscle. The critical genes were KLHL41, MYBPC2, and ERAP2. Single-gene GSEA identified the Hippo signaling pathway, basal cell carcinoma, cell adhesion molecules, and other pathways. Six miRNAs and 18 lncRNAs related to the critical genes constituted the ceRNA network and were verified using qPCR. The differential expression of KLHL41, MYBPC2, and ERAP2 at the protein level was verified using immunohistochemistry. KLHL41, MYBPC2, and ERAP2 genes are related to skin photoaging. The prediction model based on the three critical genes can indicate photoaging. These critical genes may have a role in skin photoaging by regulating cell growth, intercellular adhesion, and substance metabolism pathways.
Subject(s)
MicroRNAs , RNA, Long Noncoding , Humans , Skin , MicroRNAs/genetics , Gene Expression Profiling , Transcriptome , Gene Regulatory Networks , AminopeptidasesABSTRACT
Recently, uncontrolled actin polymerization has been recognized as an initiator of early-onset blood-brain barrier (BBB) rupture. Here, using in vitro models, we found that after oxygen-glucose deprivation/reperfusion (OGD/R), endothelial overexpression of HSPB8 suppressed aberrant actin polymerization and thus preserved the integrity of BBB. We further investigated the mechanisms of HSPB8 in the control of actin assembly. HSPB8 suppressed the RhoA/ROCK2/p-MLC signaling pathway in bEnd.3 cells and the RhoA activator abrogated the inhibitory action of HSPB8 on actin reorganization after OGD/R. In addition, endothelial autophagic flux was impaired after OGD/R. This effect was attenuated by HSPB8 overexpression. Autophagy inhibition partially reversed the effect of HSPB8 on the RhoA/ROCK2/p-MLC pathway. Taken together, the present study revealed that the restoration of autophagic flux by overexpressing HSPB8, via the inhibition of the RhoA/ROCK2/p-MLC signaling pathway, reverses the aggregation of endothelial cytoskeleton actin, eventually alleviating OGD/R-induced BBB injury.