Urolithin A Hijacks ERK1/2-ULK1 Cascade to Improve CD8+ T Cell Fitness for Antitumor Immunity.

According to the latest evidence, the microbial metabolite Urolithin A (UA), known for its role in promoting cellular health, modulates CD8+ T cell-mediated antitumor activity. However, the direct target protein of UA and its underlying mechanism remains unclear. Here, this research identifies ERK1/2 as the specific target crucial for UA-mediated CD8+ T cell activation. Even at low doses, UA markedly enhances the persistence and effector functions of primary CD8+ cytotoxic T lymphocytes (CTLs) and human chimeric antigen receptor (CAR) T cells both in vitro and in vivo. Mechanistically, UA interacts directly with ERK1/2 kinases, enhancing their activation and subsequently facilitating T cell activation by engaging ULK1. The UA-ERK1/2-ULK1 axis promotes autophagic flux in CD8+ CTLs, enhancing cellular metabolism and maintaining reactive oxygen species (ROS) levels, as evidenced by increased oxygen consumption and extracellular acidification rates. UA-treated CD8+ CTLs also display elevated ATP levels and enhanced spare respiratory capacity. Overall, UA activates ERK1/2, inducing autophagy and metabolic adaptation, showcasing its potential in tumor immunotherapy and interventions for diseases involving ERKs.

Salivary Amylase-Responsive Buccal Tablets Wipe Out Chemotherapy-Rooted Refractory Oral Mucositis.

Oral mucositis (OM) is the most common and refractory complication of cancer chemotherapy and radiotherapy, severely affecting patients' life quality, lowering treatment tolerance, and discouraging patient compliance. Current OM delivery systems mostly affect the comfort of patient use and lead to poor compliance and unsatisfactory effects. Herein, salivary amylases (SAs)-responsive buccal tablets consisting of porous manganese-substituted Prussian blue (PMPB) nanocubes (NCs), anti-inflammatory apremilast (Apr) and starch controller have been engineered. PMPB NCs with large surface area can serve as carriers to load Apr, and their multienzyme-mimicking activity enables them to scavenge reactive oxygen species (ROS), which thus synergize with Apr to mitigate inflammation. More significantly, the starch controller can respond to abundant SAs in the oral cavity and realize the cascade, continuous, and complete drug release after enzymatic decomposition, which not only aids with high tissue affinity to prolong the resistance time but also improves the comfort of use. The preclinical study reveals that contributed by the above actions, such buccal tablets mitigate inflammation, promote endothelium proliferation and migration, and accelerate wound healing for repressing chemotherapy-originated intractable OM with positive oral microenvironment and shorter recovery time, thus holding high potentials in clinical translation.

Exogenous brassinolide treatment regulates phenolic accumulation in mung bean sprouts through the modulation of sugar and energy metabolism.

BACKGROUND: The effects of exogenous brassinolide (BR) treatment (3.0 µmol L-1 ) on phenolic biosynthesis in mung bean sprouts were investigated. This investigation included the analysis of sugar content, substrates within the phenylpropane pathway, energy substances, enzymatic activity within the phenylpropane pathway, sugar metabolism and energy metabolism. RESULTS: Results showed that BR treatment significantly increased the levels of total phenolics, p-hydroxybenzoic acid, p-coumaric acid, gallic acid, fumalic acid and caffeic acid. This enhancement was accomplished through the elevation of l-phenylalanine levels and the activation of enzymes associated with the phenylpropane pathway in mung bean sprouts, including phenylalanine ammonia-lyase, cinnamate 4-hydroxylase and 4-coumarate CoA ligase. Furthermore, BR treatment induced alterations in sugar metabolism in mung bean sprouts as evidenced by the increased levels of glucose, fructose, sucrose and phosphoenolpyruvate. Moreover, increased activity was observed for enzymes linked to sucrose metabolism and glycolysis in the BR-treated group. Concurrently, BR treatment bolstered the levels of adenosine triphosphate and energy charge in mung bean sprouts, which was attributed to the activation of H+ -adenosine triphosphatase, Ca2+ -adenosine triphosphatase and succinic dehydrogenase. CONCLUSION: These results suggest that BR treatment can accelerate the accumulation of phenolic compounds in mung bean sprouts. This effect is achieved not only through the activation of the phenylpropane pathway, but also through the modulation of sugar and energy metabolism. The modulation provides ample energy and a substrate for the biosynthesis of phenolics. © 2023 Society of Chemical Industry.

Emerging Nano-/Biotechnology Drives Oncolytic Virus-Activated and Combined Cancer Immunotherapy.

Oncolytic viruses (OVs) as one promising antitumor methods have made important contributions to tumor immunotherapy, which arouse increasing attention. They provide the dual mechanisms including direct killing effect toward tumor cells and immune activation for elevating antitumor responses, which have been proved in many preclinical studies. Especially, natural or genetically modified viruses as clinical immune preparations have emerged as a new promising approach objective to oncology treatment. The approval of talimogene laherparepvec (T-VEC) by the U.S. Food and Drug Administration (FDA) for the therapy of advanced melanoma could be considered as a milestone achievement in the clinical translation of OV. In this review, we first discussed the antitumor mechanisms of OVs with an emphasis on targeting, replication, and propagation. We further outlined the state of the art of current OVs in tumor and underlined the activated biological effects especially including immunity. More significantly, the enhanced immune responses based on OVs were systematically discussed from different perspectives such as combination with immunotherapy, genetic engineering of OVs, integration with nanobiotechnology or nanoparticles, and antiviral response counteraction, where their principles were shed light on. The development of OVs in the clinics was also highlighted to analyze the actuality and concerns of different OV applications in clinical trials. At last, the future perspectives and challenges of OVs as an already widely accepted treatment means were discussed. This review will provide a systematic review and deep insight into OV development and also offer new opportunities and guidance pathways to drive the further clinical translation.

Oncolytic Impediment/Promotion Balance Disruption by Sonosensitizer-Free Nanoplatforms Unfreezes Autophagy-Induced Resistance to Sonocatalytic Therapy.

Autophagy as a double-edged sword features an oncolytic impediment/promotion balance, which manipulates tumor progression. From this perspective, a sonosensitizer-free targeting oncolytic nanoplatform (SFTON) consisting of chloroquine (CQ) and porphyrin-structured metal centers (PMCS) was engineered to break this balance for enhancing antitumor activity. Porphyrin structure retention in a ZIF-8-derived hydrophobic carbon skeleton retained high stability and high sonocatalytic activity, and the hydrophobic carbon skeleton capable of adsorbing air provided cavitation nuclei for further elevating sonocatalytic activity. More significantly, the encapsulated CQ as the autophagy inhibitor reprogrammed autophagy, terminated the autophagy-induced self-protection or self-detoxification, and unfroze the resistances to reactive oxygen species (ROS) therapy associated with ROS accumulation and ROS activity. Systematic experiments reveal the action principles and validate that the induced apoptosis and blockaded autophagosome escalation into the autolysosome were two activated pathways to magnify the antitumor sonocatalytic therapy. Contributed by these actions, the SFTON-unlocked oncolytic impediment/promotion balance disruption strategy acquired considerable antitumor outcomes in vivo and in vitro against liver tumor progression, especially after combining with AS1411-mediated active targeting. This impediment/promotion balance disruption enabled by the SFTON can serve as a general method to elevate ROS-based antitumor activity.

Emerging nanobiotechnology-encoded relaxation tuning establishes new MRI modes to localize, monitor and predict diseases.

Magnetic resonance imaging (MRI) is one of the most important techniques in the diagnosis of many diseases including cancers, where contrast agents (CAs) are usually necessary to improve its precision and sensitivity. Previous MRI CAs are confined to the signal-to-noise ratio (SNR) elevation of lesions for precisely localizing lesions. As nanobiotechnology advances, some new MRI CAs or nanobiotechnology-enabled MRI modes have been established to vary the longitudinal or transverse relaxation of CAs, which are harnessed to detect lesion targets, monitor disease evolution, predict or evaluate curative effect, etc. These distinct cases provide unexpected insights into the correlation of the design principles of these nanobiotechnologies and corresponding MRI CAs with their potential applications. In this review, first, we briefly present the principles, classifications and applications of conventional MRI CAs, and then elucidate the recent advances in relaxation tuning via the development of various nanobiotechnologies with emphasis on the design strategies of nanobiotechnology and the corresponding MRI CAs to target the tumor microenvironment (TME) and biological targets or activities in tumors or other diseases. In addition, we exemplified the advantages of these strategies in disease theranostics and explored their potential application fields. Finally, we analyzed the present limitations, potential solutions and future development direction of MRI after its combination with nanobiotechnology.

Research on the indirect antiviral function of medicinal plant ingredient quercetin against grouper iridovirus infection.

Grouper iridovirus is a devastating pathogen that belongs to the genus Ranavirus. Based on the previous results that natural ingredient quercetin isolated from Illicium verum Hook. f. could effectively inhibit Singapore grouper iridovirus (SGIV) replication, suggesting that quercetin could serve as potential antiviral agent against grouper iridovirus. To know about whether quercetin has indirect antiviral activity against SGIV, this study made the investigation in vitro and in vivo, and the potential mechanism was also explored. Pretreating the cells with quercetin (12.5 µg/mL) significantly inhibited the replication of SGIV, similar results were also confirmed in vivo. Importantly, quercetin pretreatment could induce the expression of genes involved in type I interferon (IFN) system (IFN, STAT1, PKR, MxI and ISG15) and TLR9. It suggested that quercetin exerted the indirect antiviral activity against SGIV infection through promoting the recognition of SGIV and activating the IFN pathway to establish the antiviral status of host cell. Taken together, our results shedded light on the indirect antiviral function of natural ingredient quercetin, and clearly demonstrated that natural ingredient quercetin will be an excellent potential agent against SGIV infection in grouper aquaculture.

Morphological Characterization and Transcriptional Regulation of Corolla Closure in Ipomoea purpurea.

Corolla closure protects pollen from high-temperature stress during pollen germination and fertilization in the ornamental plant morning glory (Ipomoea purpurea). However, the morphological nature of this process and the molecular events underpinning it remain largely unclear. Here, we examined the cellular and gene expression changes that occur during corolla closure in the I. purpurea. We divided the corolla closure process into eight stages (S0-S7) based on corolla morphology. During flower opening, bulliform cells appear papillate, with pigments in the adaxial epidermis of the corolla. These cells have distinct morphology from the smaller, flat cells in the abaxial epidermis in the corolla limb and intermediate of the corolla. During corolla closure, the bulliform cells of the adaxial epidermis severely collapse compared to cells on the abaxial side. Analysis of transparent tissue and cross sections revealed that acuminate veins in the corolla are composed of spiral vessels that begin to curve during corolla closure. When the acuminate veins were compromised, the corolla failed to close normally. We performed transcriptome analysis to obtain a time-course profile of gene expression during the process from the open corolla stage (S0) to semi-closure (S3). Genes that were upregulated from S0 to S1 were enriched in the polysaccharide degradation pathway, which positively regulates cell wall reorganization. Senescence-related transcription factor genes were expressed beginning at S1, leading to the activation of downstream autophagy-related genes at S2. Genes associated with peroxisomes and ubiquitin-mediated proteolysis were upregulated at S3 to enhance reactive oxygen species scavenging and protein degradation. Therefore, bulliform cells and acuminate veins play essential roles in corolla closure. Our findings provide a global understanding of the gene regulatory processes that occur during corolla closure in I. purpurea.

Reactive oxygen species scavenging and inflammation mitigation enabled by biomimetic prussian blue analogues boycott atherosclerosis.

BACKGROUND: As one typical cardiovascular disease, atherosclerosis severely endanger people' life and cause burden to people health and mentality. It has been extensively accepted that oxidative stress and inflammation closely correlate with the evolution of atherosclerotic plaques, and they directly participate in all stages of atherosclerosis. Regarding this, anti-oxidation or anti-inflammation drugs were developed to enable anti-oxidative therapy and anti-inflammation therapy against atherosclerosis. However, current drugs failed to meet clinical demands. METHODS: Nanomedicine and nanotechnology hold great potential in addressing the issue. In this report, we engineered a simvastatin (Sim)-loaded theranostic agent based on porous manganese-substituted prussian blue (PMPB) analogues. The biomimetic PMPB carrier could scavenge ROS and mitigate inflammation in vitro and in vivo. Especially after combining with Sim, the composite Sim@PMPB NC was expected to regulate the processes of atherosclerosis. As well, Mn2+ release from PMPB was expected to enhance MRI. RESULTS: The composite Sim@PMPB NC performed the best in regulating the hallmarks of atherosclerosis with above twofold decreases, typically such as oxidative stress, macrophage infiltration, plaque density, LDL internalization, fibrous cap thickness and foam cell birth, etc. Moreover, H2O2-induced Mn2+ release from PMPB NC in atherosclerotic inflammation could enhance MRI for visualizing plaques. Moreover, Sim@PMPB exhibited high biocompatibility according to references and experimental results. CONCLUSIONS: The biomimetic Sim@PMPB theranostic agent successfully stabilized atherosclerotic plaques and alleviated atherosclerosis, and also localized and magnified atherosclerosis, which enabled the monitoring of H2O2-associated atherosclerosis evolution after treatment. As well, Sim@PMPB was biocompatible, thus holding great potential in clinical translation for treating atherosclerosis.

Response of Zebrina pendula leaves to enhanced UV-B radiation.

Plants inevitably receive harmful UV-B radiation when exposed to solar energy, so they have developed a variety of strategies to protect against UV-B radiation damage during long-term evolution. In this study, Zebrina pendulaSchnizl. was used to investigate the plant defence against UV-B radiation because of its strong adaptability to sunlight changes, and the colour of its leaves changes significantly under different sunlight intensities. The experiment was carried out to study the changes of Z. pendula leaves under three light conditions: artificial daylight (control check); shading 50%; and artificial daylight + UV-B, aiming to explore the mechanism of defence against UV-B radiation by observing changes in leaf morphological structure, anthocyanin content and distribution. Results showed that the single leaf area increased but leaves became thinner, and the anthocyanin content in the epidermal cells decreased under 50% shading. In contrast, under daylight + UV-B, the single leaf area decreased but thickness increased (mainly due to the increase of the thickness of the upper epidermis and the palisade tissue), the trichomes increased. In addition, the anthocyanin content in the epidermal cells and phenylalanine ammonia-lyase (PAL) activity increased, and the leaf colour became redder, also, the photosynthetic pigment content in mesophyll cells and the biomass per unit volume increased significantly under daylight + UV-B. Thus, when UV-B radiation was enhanced, Z. pendula leaves reduced the exposure to UV-B radiation by reducing the area, and reflect some UV-B radiation by growing trichomes. The UV-B transmittance was effectively reduced by increasing the single leaf thickness and anthocyanin content to block or absorb partial UV-B. Through the above comprehensive defence strategies, Z. pendula effectively avoided the damage of UV-B radiation to mesophyll tissue.

Nanotechnology-enabled Chemodynamic Therapy and Immunotherapy.

High-level reactive oxygen species (ROS) have been reported to exert a robust anti-tumor effect by inducing cell apoptosis or necroptosis. Based on the Fenton reaction or Fenton-like reaction, a therapeutic strategy (i.e., chemodynamic therapy (CDT)) is proposed, where hydroxyl radicals (·OH) are one of the ROS that can be produced to kill tumors via the spontaneous activation by an endogenous stimulus. Moreover, high-level ROS can also facilitate tumor-associated antigen exposure, which benefits phagocytosis of corpses and debris by antigen-presenting cells (e.g., dendritic cells (DCs)) and further activates systematic immune responses. Great efforts have been made, wherein the development in the field of nanotechnology has been witnessed by the interdisciplinary communities. For providing a comprehensive understanding of CDT, state-of-theart strategies on nanotechnology-enabled CDT have been discussed in detail in this study. In particular, the combination of CDT with its augmented immunotherapy against tumors has been highlighted for overcoming the poor outcome of the mono-CDT. Moreover, the potential challenges have also been discussed.

Selection and Characterization of ssDNA Aptamers Targeting Largemouth Bass Virus Infected Cells With Antiviral Activities.

Largemouth bass virus (LMBV) is one of the most devastating viral pathogens in farmed Largemouth bass. Aptamers are novel molecule probes and have been widely applied in the field of efficient therapeutic and diagnostic agents development. LMBV-infected fathead minnow cells (LMBV-FHM) served as target cells in this study, and three DNA aptamers (LBVA1, LBVA2, and LBVA3) were generated against target cells by SELEX technology. The selected aptamers could specifically bind to LMBV-FHM cells, with rather high calculated dissociation constants (Kd) of 890.09, 517.22, and 249.31 nM for aptamers LBVA1, LBVA2, and LBVA3, respectively. Three aptamers displayed efficient antiviral activities in vitro. It indicates that the selected aptamers have great potentials in developing efficient anti-viruses treatments. The targets of aptamers LBVA1, LBVA2, and LBVA3 could be membrane proteins on host cells. The targets of aptamers (LBVA1, LBVA2, and LBVA3) come out on the cells surface at 8, 10, 8 h post-infection. As novel molecular probes for accurate recognition, aptamer LBVA3 could detect LMBV infection in vitro and in vivo, it indicates that the selected aptamers could be applied in the development of rapid detective technologies, which are characterized by high sensitivity, accuracy, and easy operation.

The inhibitory activities and antiviral mechanism of Viola philippica aqueous extracts against grouper iridovirus infection in vitro and in vivo.

Grouper iridovirus (GIV) is one of the most serious pathogens in mariculture and causes high mortality rates in cultured groupers; then, effective medicines for controlling GIV infections are urgently needed. Viola philippica is a well-known medicinal plant, and the application of V. philippica aqueous extracts against GIV infection was assessed by different methods in this study. The results showed that the working concentration of V. philippica aqueous extracts was 10 mg/ml. V. philippica aqueous extracts below 10 mg/ml have no significant cytotoxic effects on cell viability, while extracts over 15 mg/ml decreased cell viability and showed cytotoxic activity. V. philippica aqueous extracts had excellent inhibitory effects against GIV infection in vitro and in vivo. The possible antiviral mechanism of V. philippica was further analysed, which indicated that V. philippica did no damages to GIV particles, but it could disturb GIV binding, entry and replication in host cells. V. philippica had the best inhibitory effects against GIV during viral infection stage of binding and replication in host cells. Overall, the results suggest that appropriate concentration of V. philippica aqueous extracts has great antiviral effects, making it an interesting candidate for developing effective medicines for preventing and controlling GIV infection in farmed groupers.

Transcriptome Analysis to Identify the Putative Biosynthesis and Transport Genes Associated with the Medicinal Components of Achyranthes bidentata Bl.

Achyranthes bidentata is a popular perennial medicine herb used for 1000s of years in China to treat various diseases. Although this herb has multiple pharmaceutical purposes in China, no transcriptomic information has been reported for this species. In addition, the understanding of several key pathways and enzymes involved in the biosynthesis of oleanolic acid and ecdysterone, two pharmacologically active classes of metabolites and major chemical constituents of A. bidentata root extracts, is limited. The aim of the present study was to characterize the transcriptome profile of the roots and leaves of A. bidentata to uncover the biosynthetic and transport mechanisms of the active components. In this study, we identified 100,987 transcripts, with an average length of 1146.8 base pairs. A total of 31,634 (31.33%) unigenes were annotated, and 12,762 unigenes were mapped to 303 pathways according to the Kyoto Encyclopedia of Genes and Genomes pathway database. Moreover, we identified a total of 260 oleanolic acid and ecdysterone genes encoding biosynthetic enzymes. Furthermore, the key enzymes involved in the oleanolic acid and ecdysterone synthesis pathways were analyzed using quantitative real-time polymerase chain reaction, revealing that the roots expressed these enzymes to a greater extent than the leaves. In addition, we identified 85 ATP-binding cassette transporters, some of which might be involved in the translocation of secondary metabolites.

[Determination of aloin content in callus of Aloe vera var. chinensis].

The relationship between aloin accumulation of Aloe vera var. chinensis and the callus cultured by the roots, stems and leaves as explants. The aloin content in callus was determined by means of HPLC and TLC. The results showed that on the MS medium with NAA 1 mg/L + 6-BA 0.5 mg/L, the differentiation degree of the callus induced from the leaves was in the highest level, meanwhile the callus contained the most aloin. The aloin content was low in the callus from stems. There was no aloin in callus from roots. It was also found that on the MS medium with 2,4-D 1 mg/L + 6-BA 0.5 mg/L, the callus differentiation was in low level and without aloin, no matter what organs were used.