Verapamil inhibits respiratory syncytial virus infection by regulating Ca2+ influx.

AIMS: The study evaluated the antiviral effect of Verapamil against respiratory syncytial virus (RSV) and investigated its underlying mechanism. MATERIALS AND METHODS: RSV-infected BALB/c mice were treated with Verapamil. Body weight, survival rates, viral load, lung damage, inflammatory factors, and the expression of RSV fusion (F) protein were analyzed. In cellular studies, intracellular Ca2+ and viral titers were measured in the presence of Verapamil, Calcium Chloride, and EGTA. A time-of-addition assay assessed the antiviral effect of Verapamil. KEY FINDINGS: Mice infected with RSV and treated with Verapamil exhibited a significant decrease in weight loss, an increase in survival rates, and reductions in viral titers, RSV F protein expression, inflammatory responses, and lung tissue injury. Verapamil reduced intracellular calcium levels, which correlated with reduced viral titers. The addition of calcium chloride reversed the anti-viral effects mediated by Verapamil, while EGTA potentiated them. The antiviral activity of Verapamil was observed during the early phase of RSV infection, likely by blocking Ca2+ channels and inhibiting virus replication. SIGNIFICANCE: Verapamil effectively inhibits RSV infection by blocking calcium channels and reducing intracellular calcium levels, thereby impeding viral replication. Thus, Verapamil shows promise as a treatment for RSV.

Development and applications of lipid hydrophilic headgroups for nucleic acid therapy.

Nucleic acid therapy is currently the most promising method for treating tumors and genetic diseases and for preventing infectious diseases. However, the biggest obstacle to this therapy is delivery of the nucleic acids to the target site, which requires overcoming problems such as capture by the immune system, the need to penetrate biofilms, and degradation of nucleic acid performance. Designing suitable delivery vectors is key to solving these problems. Lipids-which consist of a hydrophilic headgroup, a linker, and a hydrophobic tail-are crucial components for the construction of vectors. The headgroup is particularly important because it affects the drug encapsulation rate, the vector cytotoxicity, and the transfection efficiency. Herein, we focus on various headgroup structures (tertiary amines, quaternary ammonium salts, peptides, piperazines, dendrimers, and several others), and we summarize and classify important lipid-based carriers that have been developed in recent years. We also discuss applications of cationic lipids with various headgroups for delivery of nucleic acid drugs, and we analyze how headgroup structure affects transport efficiency and carrier toxicity. Finally, we briefly describe the challenges of developing novel lipid carriers, as well as their prospects.

Blood cellular membrane-coated Au/polydopamine nanoparticle-targeted NIR-II antibacterial therapy.

Bacterial infections are the primary causes of infectious diseases in humans. In recent years, the abuse of antibiotics has led to the widespread enhancement of bacterial resistance. Concerns have been raised about the identification of a common treatment platform for bacterial infections. In this study, a composite nanomaterial was used for near-infrared II (NIR-II) photothermal antibacterial treatment. Red blood cell membrane was peeled and coated onto the surface of the Au/polydopamine nanoparticle-containing aptamer. The composite nanomaterials based on Au/polydopamine exhibit highest photothermal conversion capability. Moreover, these assembled nanoparticles can quickly enter the body's circular system with a specific capability to recognise bacteria. In vivo experiments demonstrated that the composites could kill bacteria from infected blood while significantly reducing the level of bacteria in various organs. Such assemblies offer a paradigm for the treatment of bacterial infections caused by the side effects of antibiotics.

Application of machine learning for high-throughput tumor marker screening.

High-throughput sequencing and multiomics technologies have allowed increasing numbers of biomarkers to be mined and used for disease diagnosis, risk stratification, efficacy assessment, and prognosis prediction. However, the large number and complexity of tumor markers make screening them a substantial challenge. Machine learning (ML) offers new and effective ways to solve the screening problem. ML goes beyond mere data processing and is instrumental in recognizing intricate patterns within data. ML also has a crucial role in modeling dynamic changes associated with diseases. Used together, ML techniques have been included in automatic pipelines for tumor marker screening, thereby enhancing the efficiency and accuracy of the screening process. In this review, we discuss the general processes and common ML algorithms, and highlight recent applications of ML in tumor marker screening of genomic, transcriptomic, proteomic, and metabolomic data of patients with various types of cancers. Finally, the challenges and future prospects of the application of ML in tumor therapy are discussed.

Sucrose ester embedded lipid carrier for DNA delivery.

Sucrose esters (SEs) have great potential in the field of nucleic acid delivery due to their unique physical and chemical properties and good biosafety. However, the mechanism of the effect of SEs structure on delivery efficiency has not been studied. The liposomes containing peptide lipids and SEs were constructed, and the effects of SEs on the interaction between the liposomes and DNA were studied. The addition of SEs affects the binding rate of liposomes to DNA, and the binding rate gradually decreases with the increase of SEs' carbon chain length. SEs also affect the binding site and affinity of liposomes to DNA, promoting the aggregation of lipids to form liposomes, where DNA wraps around or compresses inside the liposomes, allowing it to compress DNA without damaging the DNA structure. COL-6, which is composed of sucrose laurate, exhibits the optimal affinity for DNA, and SE promotes the formation of ordered membrane structure and enhances membrane stability, so that COL-6 exhibits a balance between rigidity and flexibility, and thus exhibits the highest delivery efficiency of DNA among these formulations. This work provides theoretical foundations for the application of SE in gene delivery and guides for the rational design of delivery systems.

ReTimeML: a retention time predictor that supports the LC-MS/MS analysis of sphingolipids.

The analysis of ceramide (Cer) and sphingomyelin (SM) lipid species using liquid chromatography-tandem mass spectrometry (LC-MS/MS) continues to present challenges as their precursor mass and fragmentation can correspond to multiple molecular arrangements. To address this constraint, we developed ReTimeML, a freeware that automates the expected retention times (RTs) for Cer and SM lipid profiles from complex chromatograms. ReTimeML works on the principle that LC-MS/MS experiments have pre-determined RTs from internal standards, calibrators or quality controls used throughout the analysis. Employed as reference RTs, ReTimeML subsequently extrapolates the RTs of unknowns using its machine-learned regression library of mass-to-charge (m/z) versus RT profiles, which does not require model retraining for adaptability on different LC-MS/MS pipelines. We validated ReTimeML RT estimations for various Cer and SM structures across different biologicals, tissues and LC-MS/MS setups, exhibiting a mean variance between 0.23 and 2.43% compared to user annotations. ReTimeML also aided the disambiguation of SM identities from isobar distributions in paired serum-cerebrospinal fluid from healthy volunteers, allowing us to identify a series of non-canonical SMs associated between the two biofluids comprised of a polyunsaturated structure that confers increased stability against catabolic clearance.

Targeting the SphK1/S1P/PFKFB3 axis suppresses hepatocellular carcinoma progression by disrupting glycolytic energy supply that drives tumor angiogenesis.

BACKGROUND: Hepatocellular carcinoma (HCC) remains a leading life-threatening health challenge worldwide, with pressing needs for novel therapeutic strategies. Sphingosine kinase 1 (SphK1), a well-established pro-cancer enzyme, is aberrantly overexpressed in a multitude of malignancies, including HCC. Our previous research has shown that genetic ablation of Sphk1 mitigates HCC progression in mice. Therefore, the development of PF-543, a highly selective SphK1 inhibitor, opens a new avenue for HCC treatment. However, the anti-cancer efficacy of PF-543 has not yet been investigated in primary cancer models in vivo, thereby limiting its further translation. METHODS: Building upon the identification of the active form of SphK1 as a viable therapeutic target in human HCC specimens, we assessed the capacity of PF-543 in suppressing tumor progression using a diethylnitrosamine-induced mouse model of primary HCC. We further delineated its underlying mechanisms in both HCC and endothelial cells. Key findings were validated in Sphk1 knockout mice and lentiviral-mediated SphK1 knockdown cells. RESULTS: SphK1 activity was found to be elevated in human HCC tissues. Administration of PF-543 effectively abrogated hepatic SphK1 activity and significantly suppressed HCC progression in diethylnitrosamine-treated mice. The primary mechanism of action was through the inhibition of tumor neovascularization, as PF-543 disrupted endothelial cell angiogenesis even in a pro-angiogenic milieu. Mechanistically, PF-543 induced proteasomal degradation of the critical glycolytic enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3, thus restricting the energy supply essential for tumor angiogenesis. These effects of PF-543 could be reversed upon S1P supplementation in an S1P receptor-dependent manner. CONCLUSIONS: This study provides the first in vivo evidence supporting the potential of PF-543 as an effective anti-HCC agent. It also uncovers previously undescribed links between the pro-cancer, pro-angiogenic and pro-glycolytic roles of the SphK1/S1P/S1P receptor axis. Importantly, unlike conventional anti-HCC drugs that target individual pro-angiogenic drivers, PF-543 impairs the PFKFB3-dictated glycolytic energy engine that fuels tumor angiogenesis, representing a novel and potentially safer therapeutic strategy for HCC.

The identification and genetic characteristics of Quang Binh virus from field-captured Culex tritaeniorhynchus (Diptera: Culicidae) from Guizhou Province, China.

BACKGROUND: Mosquitoes carry a variety of viruses that can cause disease in humans, animals and livestock. Surveys for viruses carried by wild mosquitoes can significantly contribute to surveillance efforts and early detection systems. In addition to mosquito-borne viruses, mosquitoes harbor many insect-specific viruses (ISVs). Quang Binh virus (QBV) is one such example, categorized as an ISV within the Flavivirus genus (family Flaviviridae). QBV has been specifically documented in Vietnam and China, with reports limited to several mosquito species. METHODS: The homogenate obtained from female mosquitoes was cultured on C6/36 (Aedes albopictus) and BHK-21 (baby hamster kidney) cell lines. Positive cultures were identified by reverse transcription-polymerase chain reaction (RT‒PCR) with taxon- or species-specific primers. Next-generation sequencing was employed to sequence the complete genomes of the identified positive samples. Subsequently, phylogenetic, gene homology, molecular evolutionary and genetic variation analyses were conducted. RESULT: In 2021, a total of 32,177 adult female mosquitoes were collected from 15 counties in Guizhou Province, China. The predominant mosquito species identified were Culex tritaeniorhynchus, Armigeres subalbatus and Anopheles sinensis. Among the collected mosquitoes, three positive cultures were obtained from Cx. tritaeniorhynchus pools, revealing the presence of Quang Binh virus (QBV) RNA sequences. Phylogenetic analysis indicated that the three Guizhou isolates, along with the prototype isolate from Vietnam, formed distinct branches. These branches were primarily closely related to other QBV isolates reported in China. Comparative analysis revealed a high degree of nucleotide and amino acid homology between the Guizhou isolates and both Vietnamese and other indigenous Chinese isolates. Additionally, nonsynonymous single-nucleotide variants (SNVs) were observed in these strains compared to the QBV prototype strain. CONCLUSION: This study represents the first report of QBV presences in Cx. tritaeniorhynchus mosquitoes in Guizhou Province, China. Phylogenetic tree analysis showed that the three Guizhou isolates were most closely related to the QBV genes found in China. In addition, the study of the genetic characteristics and variation of this virus provided a deeper understanding of QBV and enriched the baseline data of these insect-specific flaviviruses (ISFVs).

Development and applications of mRNA treatment based on lipid nanoparticles.

Nucleic acid-based therapies such as messenger RNA have the potential to revolutionize modern medicine and enhance the performance of existing pharmaceuticals. The key challenges of mRNA-based therapies are delivering the mRNA safely and effectively to the target tissues and cells and controlling its release from the delivery vehicle. Lipid nanoparticles (LNPs) have been widely studied as drug carriers and are considered to be state-of-the-art technology for nucleic acid delivery. In this review, we begin by presenting the advantages and mechanisms of action of mRNA therapeutics. Then we discuss the design of LNP platforms based on ionizable lipids and the applications of mRNA-LNP vaccines for prevention of infectious diseases and for treatment of cancer and various genetic diseases. Finally, we describe the challenges and future prospects of mRNA-LNP therapeutics.

An Innovative Approach to Prepare Liquid-Solid Dual-Phase Flowable Tritium Breeder with Low MHD Effect.

In present paper, a novel flowable tritium breeder is prepared by mixing the Li2TiO3 micro-powders and liquid GaInSn alloy, where GaInSn alloy is used to simulate the fluid behaviors of lithium-based liquid tritium breeder, forming a type of composite characterized by liquid-solid dual phase. In detail, the effects of the volume fraction of ceramic micro-powders on viscosity and conductivity of the composite in magnetic field are the focus. The XRD results prove that the obtained Li2TiO3 micro-powders contained Li2TiO3 phase without impurities. The results shows that once the magnetic field intensity exceeds the critical value, the viscosity of liquid GaInSn metal becomes significantly greater than that of liquid-solid dual-phase composites. Furthermore, the addition of Li2TiO3 micro-powders could effectively reduce the magneto hydro dynamic (MHD) fluid effect, and the dual-phase composites exhibit comparatively lower flow resistance under the strong magnetic field. Moreover, the conductivity of the tritium breeder composites decreases rapidly with the addition of Li2TiO3 micro-powders. The MHD pressure-drop-increasing rate decreases with the increase of viscosity, which indicates that the addition of Li2TiO3 micro-powders effectively reduces the MHD effect. The conductivity of the composites increased slightly and then remained stable after static placing for several tens of minutes. The present investigation provides a novel insight into the fabrication strategy of tritium breeder materials with low MHD effect.

Ablation of sphingosine kinase 2 suppresses fatty liver-associated hepatocellular carcinoma via downregulation of ceramide transfer protein.

Hepatocellular carcinoma (HCC) accounts for 90% of primary liver cancer, the third leading cause of cancer-associated death worldwide. With the increasing prevalence of metabolic conditions, non-alcoholic fatty liver disease (NAFLD) is emerging as the fastest-growing HCC risk factor, and it imposes an additional layer of difficulty in HCC management. Dysregulated hepatic lipids are generally believed to constitute a deleterious environment cultivating the development of NAFLD-associated HCC. However, exactly which lipids or lipid regulators drive this process remains elusive. We report herein that sphingosine kinase 2 (SphK2), a key sphingolipid metabolic enzyme, plays a critical role in NAFLD-associated HCC. Ablation of Sphk2 suppressed HCC development in NAFLD livers via inhibition of hepatocyte proliferation both in vivo and in vitro. Mechanistically, SphK2 deficiency led to downregulation of ceramide transfer protein (CERT) that, in turn, decreased the ratio of pro-cancer sphingomyelin (SM) to anti-cancer ceramide. Overexpression of CERT restored hepatocyte proliferation, colony growth and cell cycle progression. In conclusion, the current study demonstrates that SphK2 is an essential lipid regulator in NAFLD-associated HCC, providing experimental evidence to support clinical trials of SphK2 inhibitors as systemic therapies against HCC.

Aire and Fezf2, two regulators in medullary thymic epithelial cells, control autoimmune diseases by regulating TSAs: Partner or complementer?

The expression of tissue-specific antigens (TSAs) in medullary thymic epithelial cells (mTECs) is believed to be responsible for the elimination of autoreactive T cells, a critical process in the maintenance of central immune tolerance. The transcription factor autoimmune regulator (Aire) and FEZ family zinc finger 2(Fezf2) play an essential role in driving the expression of TSAs in mTECs, while their deficiency in humans and mice causes a range of autoimmune manifestations, such as type 1 diabetes, Sjögren's syndrome and rheumatoid arthritis. However, because of their regulatory mechanisms, the expression profile of TSAs and their relationship with special autoimmune diseases are still in dispute. In this review, we compare the roles of Aire and Fezf2 in regulating TSAs, with an emphasis on their molecular mechanisms in autoimmune diseases, which provides the foundation for devising improved diagnostic and therapeutic approaches for patients.

Colorimetric sensing of glucose and GSH using core-shell Cu/Au nanoparticles with peroxidase mimicking activity.

The catalytic properties of bimetallic nanoparticles have been widely studied by researchers in many fields. In this paper, core-shell Cu/Au nanoparticles (Cu/Au NPs) were synthesized by a simple and mild one-pot method, and their peroxidase activity was proved by catalyzing the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) with color change to blue. The change of solution color and absorbance strongly depends on the concentration of H2O2, so it can be used for direct detection of H2O2 and indirect detection of glucose. What's more, GSH can efficiently react with the hydroxyl radicals from H2O2 catalyzed by core-shell Cu/Au NPs to inhibit the production of ox-TMB. Thus, the concentration of GSH can be determined by the decrease in the absorbance of the solution at 652 nm. The results showed that our proposed strategy had good detection range and detection limit for the detection of glucose and GSH. This method has been used in the detection of practical samples and has great application potential in environmental monitoring and clinical diagnosis.

Polyoxometalates-graphene nanocomposites modified electrode for electro-sensing detection of Sudan I in food.

Sudan I, a lipophilic azo dye -dye, is desirable and urgent to be accurate detected due to its increasing levels and high toxicity in food and environmental monitoring and analysis. Herein, a sensitive electrochemical sensor for Sudan I was established based on a new K10P2W18Fe4(H2O)2O68 functionalized carbon nanomaterials (Fe4P2W18-GNPS). The electrode modified nanocomposite, Fe4P2W18-GNPS, was successfully fabricated and characterized by FTIR, SEM and UV-vis. The effective combination of Fe4P2W18 and graphene exhibited high electrocatalytic activity towards the oxidation of Sudan I, promote charge transfer, and more sensing sites. Under optimized experimental conditions, the proposed differential pulse voltammetry (DPV) showed excellent analytical performances for Sudan I with the limit of detection (LOD) of 5 nM (S/N = 3), the sensitivity of 13.10 µA·µM-1cm-2 at the 0.005-2 µM and 0.39 µA·µM-1cm-2 at 10-200 µM. The stability and reproducibility make the electrochemical sensor suitable for detecting the Sudan I in food.

Cu2+ modified Zr-based metal organic framework-CTAB-graphene for sensitive electrochemical detection of sunset yellow.

A sensitive electrochemical sensor for sunset yellow (SY) was constructed based on cetyltrimethylammonium bromide (CTAB) functionalized graphene (Gr) and Cu/Zr-MOF electrode modified materials. The CTAB-Gr-Cu/Zr-MOF composites were synthesized by using a mild method and characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and EDX spectrum. The combination of Cu/Zr-MOF and graphene exhibited synergetic effect of the strong accumulation efficiency, fast electron transfer rate and more sensing sites towards the oxidation of SY. The new modified materials remarkably increased the electrochemical response of SY to 6.53-fold when comparing with bare electrode. Under the optimized conditions, the oxidation peak currents of SY had a linear relationship with its concentration in a wide range from 0.10 to 8.00 µM and 40.00-1000.00 µM, and the limit of detection was 6.68 nM (S/N = 3). The electrochemical method shows high sensitivity, stability, reproducibility and is successfully applied in the determination of SY in soft drinks.

Recent Advances in Polyoxometalates with Enzyme-like Characteristics for Analytical Applications.

Artificial enzymes based on inorganic solids with both enzyme-mimetic activities and the special material features has been a promising candidate to overcome many deleterious effects of native enzymes in analytical applications. Polyoxometalates (POMs) are an importance class of molecular metal-oxygen anionic clusters. Their outstanding physicochemical properties, versatility and potential applications in energy conversion, magnetism, catalysis, molecular electronics and biomedicine have long been studied. However, the analytical applications of them is limited. Recently, the intrinsic enzymatic activities of POMs have also been found and become an area of growing interest. In this review, along with other reports, we aimed to classify the enzymatic activity of POMs, summarize the construction of POMs-based enzymes, and survey their recent advances in analytical fields. Finally, the current challenges and trends of the polyoxometalates with enzymatic activity in future chemo-/bio-sensing applications are briefly discussed.

Review on Preparation Technology and Properties of Refractory High Entropy Alloys.

Refractory high entropy alloys have broad application prospects due to their excellent comprehensive properties in high temperature environments, and they have been widely implemented in many complex working conditions. According to the latest research reports, the preparation technology of bulk and coating refractory high entropy alloys are summarized, and the advantages and disadvantages of each preparation technology are analyzed. In addition, the properties of refractory high entropy alloys, such as mechanical properties, wear resistance, corrosion resistance, oxidation resistance, and radiation resistance are reviewed. The existing scientific problems of refractory high entropy alloys, at present, are put forward, which provide reference for the development and application of refractory high entropy alloys in the future, especially for plasma-facing materials in nuclear fusion reactors.

A nanotherapy responsive to the inflammatory microenvironment for the dual-targeted treatment of atherosclerosis.

Atherosclerosis remains the main cause of death and disability, as well as a leading cause of coronary arterial disease. Inflammation is one of the pathogenic factors of arteriosclerosis; however, the current treatments based on lowering the level of inflammation in the plaque tissue of patients with atherosclerosis are not clinically used. Herein, we hypothesize that αvß3 receptor affinity and low pH sensitivity may be regarded as a valid therapeutic strategy for targeting sites of atherosclerosis according to the microenvironments of inflammation. To prove this tentative hypothesis, an acid-labile material polyketal named PK3 was synthesized, and the cRGDfc peptide was used to modify nanoparticles composed of poly(lactide-co-glycolide) (PLGA), lecithin, and PK3, loaded with the anti-atherosclerotic drug rapamycin (RAP). The nanoparticles were prepared using an O/W method and then characterized, which showed an appropriate particle size and fulfilling responsive behaviors. In vitro release studies and stability tests showed that these nanoparticles can be effectively internalized by human umbilical vein endothelial cells (HUVEC), and also show a good in vitro anti-inflammatory effect. After intravenous (i.v.) injection, RGD targeted by pH-responsive nanotherapy (RAP-Nps-RGD) may be accumulated at the plaque site in ApoE-/- mice with atherosclerosis and can effectively attenuate plaque progression compared to other formulations. Moreover, its good safety profile and biocompatibility have been revealed in both in vitro and in vivo estimations. Accordingly, the prospect of nanoparticles responsive to the inflammatory microenvironment for preventing atherosclerotic through inflammation modulation provides great feasibility for the administration of alternate drug molecules to inflamed sites to slow down the process of arteriosclerosis.

A Multifunctional Janus Electrospun Nanofiber Dressing with Biofluid Draining, Monitoring, and Antibacterial Properties for Wound Healing.

Wound healing greatly affects patients' health and produces medical burden. Therefore, we developed a multifunctional electrospun nanofiber dressing, which can inhibit methicillin-resistant Staphylococcus aureus (MRSA), drain excessive biofluid to promote wound healing, and simultaneously monitor wound pH level. The polyoxometalate (α-K6P2W18O62·14H2O, P2W18) and oxacillin (OXA) are encapsulated in hydrophobic polylactide (PLA) nanofiber to synergistically inhibit MRSA. The phenol red (PSP) is encapsulated in hydrophilic polyacrylonitrile (PAN) nanofiber to sensitively indicate wound pH in situ. The PSP/PAN nanofiber is directly electrospun on the patterning OXA/P2W18/PLA nanofiber layer to form a Janus dressing. By taking advantage of the wettability difference between the two layers, the excess biofluid can be drained away from the wound. In addition, the Janus dressing exhibits good biocompatibility and accelerates wound healing via its antimicrobial activity and skin repairing function. This multifunctional Janus electrospun nanofiber dressing would be beneficial for wound management and treatment.

Multienzymatic Antioxidant Activity of Manganese-Based Nanoparticles for Protection against Oxidative Cell Damage.

Oxidative stress is related to many diseases, but available clinical treatment methods are currently limited. Exploitation of enzyme-mimicking nanomaterials (nanozymes) is a promising way for scavenging reactive oxygen species (ROS) and treatment of ROS-related diseases. Herein, the catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) mimicking activities are expressed by MnO2 nanoparticles (MnO2-BSA NPs) coated with BSA. Effective •OH removal activity is also expressed by MnO2-BSA NPs at neutral pH. Apoptosis inhibition and ROS scavenging capabilities of MnO2-BSA NPs are evident on the H2O2-exposed BEAS-2B cells line. Western blot analysis indicates that MnO2-BSA NPs inhibit H2O2-induced apoptosis by mediating the expression of apoptosis-related proteins.