ABSTRACT
African swine fever (ASF), caused by the African swine fever virus (ASFV), is a highly infectious disease afflicting domestic pigs and wild boars. It exhibits an alarming acute infection fatality rate of up to 100%. Regrettably, no commercial vaccines or specific drugs for combating this disease are currently available. This study evaluated the anti-ASFV activities in porcine alveolar macrophages, 3D4/21 cells, and PK-15 cells of four bis-benzylisoquinoline alkaloids (BBAs): cepharanthine (CEP), tetrandrine, fangchinoline, and iso-tetrandrine. Furthermore, we demonstrated that CEP, which exhibited the highest selectivity index (SI = 81.31), alkalized late endosomes/lysosomes, hindered ASFV endosomal transport, disrupted virus uncoating signals, and thereby inhibited ASFV internalization. Additionally, CEP disrupted ASFV DNA synthesis, leading to the inhibition of viral replication. Moreover, berbamine was labeled with NBD to synthesize a fluorescent probe to study the cellular location of these BBAs. By co-staining with Lyso-Tracker and lysosome-associated membrane protein 1, we demonstrated that BBAs target the endolysosomal compartments for the first time. Our data together indicated that BBAs are a class of natural products with significant inhibitory effects against ASFV infection. These findings suggest their potential efficacy as agents for the prevention and control of ASF, offering valuable references for the identification of potential drug targets.IMPORTANCEThe urgency and severity of African swine fever (ASF) underscore the critical need for effective interventions against this highly infectious disease, which poses a grave threat to domestic pigs and wild boars. Our study reveals the potent anti-African swine fever virus (ASFV) efficacy of bis-benzylisoquinoline alkaloids (BBAs), particularly evident in the absence of progeny virus production under a 5 µM concentration treatment. The structural similarity among cepharanthine, tetrandrine, fangchinoline, and iso-tetrandrine, coupled with their analogous inhibitory stages and comparable selectivity indexes, strongly suggests a shared antiviral mechanism within this drug category. Further investigation revealed that BBAs localize to lysosomes and inhibit the internalization and replication of ASFV by disrupting the endosomal/lysosomal function. These collective results have profound implications for ASF prevention and control, suggesting the potential of the investigated agents as prophylactic and therapeutic measures. Furthermore, our study offers crucial insights into identifying drug targets and laying the groundwork for innovative interventions.
Subject(s)
African Swine Fever Virus , Antiviral Agents , Benzylisoquinolines , Endosomes , Lysosomes , Virus Internalization , Virus Replication , Animals , African Swine Fever Virus/drug effects , African Swine Fever Virus/physiology , Virus Internalization/drug effects , Benzylisoquinolines/pharmacology , Virus Replication/drug effects , Lysosomes/drug effects , Lysosomes/metabolism , Lysosomes/virology , Swine , Endosomes/metabolism , Endosomes/drug effects , Endosomes/virology , Antiviral Agents/pharmacology , Cell Line , African Swine Fever/virology , African Swine Fever/drug therapy , African Swine Fever/metabolism , Guanine/analogs & derivatives , Guanine/pharmacology , Alkaloids/pharmacology , Macrophages, Alveolar/virology , Macrophages, Alveolar/drug effects , BenzodioxolesABSTRACT
Atmospheric water harvesting (AWH) is considered a promising strategy for sustainable freshwater production in landlocked and arid regions. Hygroscopic salt-based composite sorbents have attracted widespread attention for their water harvesting performance, but suffer from aggregation and leakage issues due to the salting-out effect. In this study, we synthesized a PML hydrogel composite by incorporating zwitterionic hydrogel (PDMAPS) and MIL-101(Cr) as a host for LiCl. The PML hydrogel was characterized using various techniques including X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared (FTIR), and thermogravimetric analysis (TGA). The swelling properties and water vapor adsorption-desorption properties of the PML hydrogel were also assessed. The results demonstrate that the MIL-101(Cr) was uniformly embedded into PDMAP hydrogel, and the PML hydrogel exhibits a swelling ratio of 2.29 due to the salting-in behavior. The PML hydrogel exhibited exceptional water vapor sorption capacity of 0.614 g/g at 298 K, RH = 40% and 1.827 g/g at 298 K, RH = 90%. It reached 80% of its saturated adsorption capacity within 117 and 149 min at 298 K, RH = 30% and 90%, respectively. Additionally, the PML hydrogel showed excellent reversibility in terms of water vapor adsorption after ten consecutive cycles of adsorption-desorption. The remarkable adsorption capacity, favorable adsorption-desorption rate, and regeneration stability make the PML hydrogel a potential candidate for AWH. This polymer-MOF synergistic strategy for immobilization of LiCl in this work offers new insights into designing advanced materials for AWH.
ABSTRACT
In China, acquiring firms are increasingly focused on the immediate financial returns that digital mergers and acquisitions (DM&A) can help them achieve in the stock market, but there is little literature that examines which acquiring firms achieve greater returns. Based on signaling theory, this study conceptualizes DM&A announcements as signals released by the acquiring firms to the stock market and explores the factors influencing the Chinese stock market's reaction to such signals. This research empirically investigates potential influencing factors using a short-term event methodology together with regression analysis based on the data collected in China's Shanghai and Shenzhen stock markets during 2012-2021. The research finds that the Chinese stock market reacts more positively to DM&A announcements for acquiring firms with high executive shareholdings, high executive openness, strong digital innovation capabilities, and in regions with higher levels of investor protection. This study is the first attempt to explore the factors influencing the stock market's response to DM&A in the Chinese context.
ABSTRACT
Chronic lumbar and back pain caused by degenerative vertebral endplates presents a challenging issue for patients and clinicians. As a new minimally invasive spinal treatment method, radiofrequency ablation of vertebral basal nerve in bone can denature the corresponding vertebral basal nerve through radiofrequency ablation of degenerative vertebral endplate. It blocks the nociceptive signal transmission of the vertebral base nerve, thereby alleviating the symptoms of low back pain caused by the degenerative vertebral endplate. At present, many foreign articles have reported the operation principle, operation method, clinical efficacy and related complications of radiofrequency ablation of the vertebral basal nerve. The main purpose of this paper is to conduct a comprehensive analysis of the current relevant research, and provide a reference for the follow-up clinical research.
Subject(s)
Radiofrequency Ablation , Humans , Radiofrequency Ablation/methods , Low Back Pain/surgery , Lumbar Vertebrae/surgery , Spinal Nerves/surgeryABSTRACT
Osteosarcoma (OS) is a highly malignant primary bone neoplasm that is the leading cause of cancerassociated death in young people. GNE477 belongs to the second generation of mTOR inhibitors and possesses promising potential in the treatment of OS but dose tolerance and drug toxicity limit its development and utilization. The present study aimed to prepare a novel H2O2 stimulusresponsive dodecanoic acid (DA)phenylborate esterdextran (DABDEX) polymeric micelle delivery system for GNE477 and evaluate its efficacy. The polymer micelles were characterized by morphology, size and critical micelle concentration. The GNE477 loaded DABDEX (GNE477@DBD) tumortargeting drug delivery system was established and the release of GNE477 was measured. The cellular uptake of GNE477@DBD by three OS cell lines (MG63, U2OS and 143B cells) was analyzed utilizing a fluorescent tracer technique. The hydroxylated DAB was successfully grafted onto dextran at a grafting rate of 3%, suitable for forming amphiphilic micelles. Following exposure to H2O2, the DABDEX micelles ruptured and released the drug rapidly, leading to increased uptake of GNE477@DBD by cells with sustained release of GNE477. The in vitro experiments, including MTT assay, flow cytometry, western blotting and RTqPCR, demonstrated that GNE477@DBD inhibited tumor cell viability, arrested cell cycle in G1 phase, induced apoptosis and blocked the PI3K/Akt/mTOR cascade response. In vivo, through the observation of mice tumor growth and the results of H&E staining, the GNE477@DBD group exhibited more positive therapeutic outcomes than the free drug group with almost no adverse effects on other organs. In conclusion, H2O2responsive DABDEX presents a promising delivery system for hydrophobic antitumor drugs for OS therapy.
Subject(s)
Dextrans , Hydrogen Peroxide , Lauric Acids , Micelles , Osteosarcoma , Animals , Humans , Osteosarcoma/drug therapy , Osteosarcoma/pathology , Cell Line, Tumor , Dextrans/chemistry , Mice , Lauric Acids/chemistry , Lauric Acids/pharmacology , Apoptosis/drug effects , Polymers/chemistry , Polymers/pharmacology , Xenograft Model Antitumor Assays , Bone Neoplasms/drug therapy , Bone Neoplasms/pathology , Mice, Nude , Antineoplastic Agents/pharmacology , Antineoplastic Agents/chemistry , Mice, Inbred BALB C , Male , TOR Serine-Threonine Kinases/metabolismABSTRACT
The NACHT, LRR, and PYD domains-containing protein 3 (NLRP3) inflammasome has been involved in the pathogenesis of various chronic liver diseases. However, its role in hepatitis B virus (HBV)-associated hepatitis remains unknown. Here we demonstrate the synergistic effect of HBV with potential intrahepatic danger signals on NLRP3 inflammasome activation. HBV exposure at the appropriate temporal points enhances potassium efflux-dependent NLRP3 inflammasome activation in macrophages and also increases NLRP3 inflammasome-mediated inflammation in HBV-transgenic mouse model. HBV-mediated synergism with intrahepatic signals represented by ATP molecules on NLRP3 activation was observed via relevance analysis, confocal microscopy, and co-immunoprecipitation, and its effector cytokines exhibit positive associations with hepatic inflammation in patients with severe hepatitis B. Furthermore, the synergism of HBV on NLRP3 inflammasome activation owes to increased sodium influx into macrophages. Our data demonstrate that HBV contributes to hepatic inflammation via sodium influx-dependent synergistic activation of NLRP3 inflammasome, which provides a deeper understanding of immune pathogenesis in HBV-associated hepatitis.
ABSTRACT
CRISPR/Cas-based transcriptional activators can be enhanced by intrinsically disordered regions (IDRs). However, the underlying mechanisms are still debatable. Here, we examine 12 well-known IDRs by fusing them to the dCas9-VP64 activator, of which only seven can augment activation, albeit independently of their phase separation capabilities. Moreover, modular domains (MDs), another class of multivalent molecules, though ineffective in enhancing dCas9-VP64 activity on their own, show substantial enhancement in transcriptional activation when combined with dCas9-VP64-IDR. By varying the number of gRNA binding sites and fusing dCas9-VP64 with different IDRs/MDs, we uncover that optimal, rather than maximal, cis-trans cooperativity enables the most robust activation. Finally, targeting promoter-enhancer pairs yields synergistic effects, which can be further amplified via enhancing chromatin interactions. Overall, our study develops a versatile platform for efficient gene activation and sheds important insights into CRIPSR-based transcriptional activators enhanced with multivalent molecules.
Subject(s)
CRISPR-Cas Systems , Transcriptional Activation , Humans , Promoter Regions, Genetic , RNA, Guide, CRISPR-Cas Systems/genetics , RNA, Guide, CRISPR-Cas Systems/metabolism , HEK293 Cells , Binding Sites , Chromatin/metabolism , Clustered Regularly Interspaced Short Palindromic Repeats , Enhancer Elements, GeneticABSTRACT
The three-dimensional (3D) organization of chromatin within the nucleus is crucial for gene regulation. However, the 3D architectural features that coordinate the activation of an entire chromosome remain largely unknown. We introduce an omics method, RNA-associated chromatin DNA-DNA interactions, that integrates RNA polymerase II (RNAPII)-mediated regulome with stochastic optical reconstruction microscopy to investigate the landscape of noncoding RNA roX2-associated chromatin topology for gene equalization to achieve dosage compensation. Our findings reveal that roX2 anchors to the target gene transcription end sites (TESs) and spreads in a distinctive boot-shaped configuration, promoting a more open chromatin state for hyperactivation. Furthermore, roX2 arches TES to transcription start sites to enhance transcriptional loops, potentially facilitating RNAPII convoying and connecting proximal promoter-promoter transcriptional hubs for synergistic gene regulation. These TESs cluster as roX2 compartments, surrounded by inactive domains for coactivation of multiple genes within the roX2 territory. In addition, roX2 structures gradually form and scaffold for stepwise coactivation in dosage compensation.