Spatially exploring RNA biology in archival formalin-fixed paraffin-embedded tissues.

The capability to spatially explore RNA biology in formalin-fixed paraffin-embedded (FFPE) tissues holds transformative potential for histopathology research. Here, we present pathology-compatible deterministic barcoding in tissue (Patho-DBiT) by combining in situ polyadenylation and computational innovation for spatial whole transcriptome sequencing, tailored to probe the diverse RNA species in clinically archived FFPE samples. It permits spatial co-profiling of gene expression and RNA processing, unveiling region-specific splicing isoforms, and high-sensitivity transcriptomic mapping of clinical tumor FFPE tissues stored for 5 years. Furthermore, genome-wide single-nucleotide RNA variants can be captured to distinguish malignant subclones from non-malignant cells in human lymphomas. Patho-DBiT also maps microRNA regulatory networks and RNA splicing dynamics, decoding their roles in spatial tumorigenesis. Single-cell level Patho-DBiT dissects the spatiotemporal cellular dynamics driving tumor clonal architecture and progression. Patho-DBiT stands poised as a valuable platform to unravel rich RNA biology in FFPE tissues to aid in clinical pathology evaluation.

Single-cell CAR T atlas reveals type 2 function in 8-year leukaemia remission.

Despite a high response rate in chimeric antigen receptor (CAR) T cell therapy for acute lymphocytic leukaemia (ALL)1-3, approximately 50% of patients relapse within the first year4-6, representing an urgent question to address in the next stage of cellular immunotherapy. Here, to investigate the molecular determinants of ultralong CAR T cell persistence, we obtained a single-cell multi-omics atlas from 695,819 pre-infusion CAR T cells at the basal level or after CAR-specific stimulation from 82 paediatric patients with ALL enrolled in the first two CAR T ALL clinical trials and 6 healthy donors. We identified that elevated type 2 functionality in CAR T infusion products is significantly associated with patients maintaining a median B cell aplasia duration of 8.4 years. Analysis of ligand-receptor interactions revealed that type 2 cells regulate a dysfunctional subset to maintain whole-population homeostasis, and the addition of IL-4 during antigen-specific activation alleviates CAR T cell dysfunction while enhancing fitness at both transcriptomic and epigenomic levels. Serial proteomic profiling of sera after treatment revealed a higher level of circulating type 2 cytokines in 5-year or 8-year relapse-free responders. In a leukaemic mouse model, type 2high CAR T cell products demonstrated superior expansion and antitumour activity, particularly after leukaemia rechallenge. Restoring antitumour efficacy in type 2low CAR T cells was attainable by enhancing their type 2 functionality, either through incorporating IL-4 into the manufacturing process or by priming manufactured CAR T products with IL-4 before infusion. Our findings provide insights into the mediators of durable CAR T therapy response and suggest potential therapeutic strategies to sustain long-term remission by boosting type 2 functionality in CAR T cells.

The type 2 cytokine Fc-IL-4 revitalizes exhausted CD8+ T cells against cancer.

Current cancer immunotherapy predominately focuses on eliciting type 1 immune responses fighting cancer; however, long-term complete remission remains uncommon1,2. A pivotal question arises as to whether type 2 immunity can be orchestrated alongside type 1-centric immunotherapy to achieve enduring response against cancer3,4. Here we show that an interleukin-4 fusion protein (Fc-IL-4), a typical type 2 cytokine, directly acts on CD8+ T cells and enriches functional terminally exhausted CD8+ T (CD8+ TTE) cells in the tumour. Consequently, Fc-IL-4 enhances antitumour efficacy of type 1 immunity-centric adoptive T cell transfer or immune checkpoint blockade therapies and induces durable remission across several syngeneic and xenograft tumour models. Mechanistically, we discovered that Fc-IL-4 signals through both signal transducer and activator of transcription 6 (STAT6) and mammalian target of rapamycin (mTOR) pathways, augmenting the glycolytic metabolism and the nicotinamide adenine dinucleotide (NAD) concentration of CD8+ TTE cells in a lactate dehydrogenase A-dependent manner. The metabolic modulation mediated by Fc-IL-4 is indispensable for reinvigorating intratumoural CD8+ TTE cells. These findings underscore Fc-IL-4 as a potent type 2 cytokine-based immunotherapy that synergizes effectively with type 1 immunity to elicit long-lasting responses against cancer. Our study not only sheds light on the synergy between these two types of immune responses, but also unveils an innovative strategy for advancing next-generation cancer immunotherapy by integrating type 2 immune factors.

Efficient Sodium Storage in Cu1.96S@NC Anode Achieved by Robust S─C Bonds and Current Collector Self-Induced Forming Cu2S Quantum Dots.

Transition metal sulfides are investigation hotspots of anode material for sodium-ion batteries (SIBs) due to their structural diversity and high storage capacity. However, they are still plagued by inevitable volume expansion during sodiation/desodiation and an unclear energy storage mechanism. Herein, a one-step sulfidation-carbonization strategy is proposed for in situ confined growth of Cu1.96S nanoparticles in nitrogen-doped carbon (Cu1.96S@NC) using octahedral metal-organic framework (Cu-BTC) as a precursor and investigate the driving effect of Cu current collector on its sodium storage. The generation of S─C bonds in Cu1.96S@NC avoids the volume change and structural collapse of Cu1.96S nanoparticles during the cycling process and improves the adsorption and transport capacity of the material for Na+. More exciting, the Cu species in the Cu current collector are self-induced forming Cu2S quantum dots to enter the original anode material during the initial few charging and discharging cycles, which unique small-size effect and abundant edge-active sites enhance the energy storage capacity of Cu1.96S. Thus, the Cu1.96S@NC exhibits a superior first discharge capacity of 608.56 mAh g-1 at 0.2 A g-1 with an initial Coulomb efficiency (ICE) of 75.4%, as well as provides excellent rate performance and long cycle durability up to 2000 cycles.

Applications of MRI in Schizophrenia: Current Progress in Establishing Clinical Utility.

Schizophrenia is a severe mental illness that significantly impacts the lives of affected individuals and with increasing mortality rates. Early detection and intervention are crucial for improving outcomes but the lack of validated biomarkers poses great challenges in such efforts. The use of magnetic resonance imaging (MRI) in schizophrenia enables the investigation of the disorder's etiological and neuropathological substrates in vivo. After decades of research, promising findings of MRI have been shown to aid in screening high-risk individuals and predicting illness onset, and predicting symptoms and treatment outcomes of schizophrenia. The integration of machine learning and deep learning techniques makes it possible to develop intelligent diagnostic and prognostic tools with extracted or selected imaging features. In this review, we aimed to provide an overview of current progress and prospects in establishing clinical utility of MRI in schizophrenia. We first provided an overview of MRI findings of brain abnormalities that might underpin the symptoms or treatment response process in schizophrenia patients. Then, we summarized the ongoing efforts in the computer-aided utility of MRI in schizophrenia and discussed the gap between MRI research findings and real-world applications. Finally, promising pathways to promote clinical translation were provided. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY: Stage 3.

Patumantanes A-D, seco-Polycyclic Polyprenylated Acylphloroglucinols with Diverse Carbon Skeletons from Hypericum patulum.

Patumantanes A-D (1-4), four new seco-polycyclic polyprenylated acylphloroglucinols (PPAPs) were isolated from Hypericum patulum. Patumantane A (1) was an unprecedented 1,2-seco-homoadamantane-type PPAP bearing a new 3,7-dioxatetracyclo[7.7.0.01,6.111,15]heptadecane architecture based on a 6/7/5/6 ring system. Patumantane B (2) was a unique 1,9-seco-adamantane-type PPAP with a tricyclo[4.4.4.0.02,12]tridecane core formed by a 6/6/6 carbon skeleton, and the further breakage between C-5 and C-9 decorated patumantane C (3) with the 9-nor-adamantane skeleton. More importantly, compounds 2 and 3 exhibited moderate immunosuppressive activity on Con A-induced T-lymphocyte proliferation in vitro, with IC50 values of 5.6 ± 1.2 and 11.2 ± 1.2 µM, respectively.

Discovery of adamantane-type polycyclic polyprenylated acylphloroglucinols that can prevent concanavalin A-induced autoimmune hepatitis in mice.

Hyperadamans A-G (1-7), seven new adamantane type polycyclic polyprenylated acylphloroglucinols (PPAPs), were isolated from Hypericum wilsonii N. Robson. Structurally, 1-4 were the first adamantanes bearing an unusual 2,7-dioxabicyclo-[2.2.1]-heptane fragment, and compound 5 was the first adamantane with a rare 1,6-dioxaspiro[4.4]nonane section. Importantly, 1-7 exhibited significant immunosuppressive activity on Con A-induced T-lymphocyte proliferation in vitro, with IC50 values ranging from 3.97 ± 0.10 to 18.12 ± 1.07 µM. Pretreatment with 1 in Con A-challenged autoimmune hepatitis mice could dramatically ameliorate the levels of hepatic injury indexes (ALT and AST) and reduce the product of proinflammatory cytokines (COX-2, IL-6, IL-1ß, IL-18, IL-23A and TNF-α). Furthermore, the protective effect of 1 on the Con A-induced liver injury was corroborated by the histological analysis and the immunohistochemistry.

Radiomic features of gray matter in never-treated first-episode schizophrenia.

Alterations of radiomic features (RFs) in gray matter are observed in schizophrenia, of which the results may be limited by small study samples and confounding effects of drug therapies. We tested for RFs alterations of gray matter in never-treated first-episode schizophrenia (NT-FES) patients and examined their associations with known gene expression profiles. RFs were examined in the first sample with 197 NT-FES and 178 healthy controls (HCs) and validated in the second independent sample (90 NT-FES and 74 HCs). One-year follow-up data were available from 87 patients to determine whether RFs were associated with treatment outcomes. Associations between identified RFs in NT-FES and gene expression profiles were evaluated. NT-FES exhibited alterations of 30 RFs, with the greatest involvement of microstructural heterogeneity followed by measures of brain region shape. The identified RFs were mainly located in the central executive network, frontal-temporal network, and limbic system. Two baseline RFs with the involvement of microstructural heterogeneity predicted treatment response with moderate accuracy (78% for the first sample, 70% for the second sample). Exploratory analyses indicated that RF alterations were spatially related to the expression of schizophrenia risk genes. In summary, the present findings link brain abnormalities in schizophrenia with molecular features and treatment response.

First Report of Root Collar Canker Caused by Neoscytalidium dimidiatum on Jacaranda mimosifolia in China.

Jacaranda mimosifolia is widely cultivated as a garden ornamental tree. In July 2023, an unknown root collar canker of J. mimosifolia was discovered in green belts of Qingxiu District, Nanning, China, with a 8% incidence rate. Crowns of affected trees ranged from reddish brown leaves to deciduous or dead. Root collar tissue became necrotic matched by underbark dark brown lesions with irregular margins, and rotted at last. Six diseased plants distributed within 3000 m2 were choosed, and 24 root collar tissues were surface sterilized and placed on potato dextrose agar (PDA) plates to incubate at 28℃ for 3 to 5 days. Same colonies were consistently isolated from 18 tissues, and three isolates (M3-B1-1, M3-B1-2 and M3-B1-3) were purified for morphological and molecular determination. These isolates formed colonies with lush aerial mycelia rapidly, which covered a 90 mm plate in 72h. The colonies were initially white, then grayish-green to black. Arthrospores were colourless to light brown, short columnar, aseptate, truncate base, averaging 12.1±2.5 µm × 3.4±0.7 µm, sometimes formed arthric chains. Chlamydospores were dark brown, round or oval, aseptate, averaging 8.7±1.6 µm × 5.0±0.9 µm. Mature pycnidia and conidia produced for about 50 days on oatmeal agar medium (OMA), and conidia were colorless, oblong, aseptate, averaging 11.2±1.2 µm × 6.0±1.4 µm. These morphological characteristics were consistent with the description of Neoscytalidium dimidiatum (Penz.) Crous & Slippers (Crous et al. 2006). Genomic DNA was extracted from three isolates. The partial ITS region, TUB2 and TEF1-α genes were amplified (White et al., 1990; Glass and Donaldson 1995; Carbone and Kohn 1999). The sequences were deposited in GenBank (ITS: PP939650-PP939652; TUB2: PP942728-PP942730; TEF1-α: PP942731-PP942733). Blastn analysis revealed that ITS sequences of three isolates showed 99.8%, 100%, 100% identity (506 bp out of 507 bp, 507 bp out of 507 bp, 507 bp out of 507 bp) to N. dimidiatum C21 (KX447539), the TUB2 sequences showed 100% identity (436 bp out of 436 bp, 437 bp out of 437 bp, 437 bp out of 437 bp) to N.dimidiatum LNeo (ON099066), and the TEF1-α sequences showed 99.64% identity (276 bp out of 277 bp) to N.dimidiatum ARM230 (MK495384), respectively. Phylogenetic analysis based on concatenated ITS, TUB2 and TEF1-α sequences showed that three isolates were clustered into the same clade as N. dimidiatum. To fulfill Koch's postulates, pathogenicity of these isolates was tested on healthy two-year-old J. mimosifolia trees. Stem and root collar were wounded and placed mycelial plugs (8mm), and the inoculation sites were wrapped with parafilm or covered with nursery substrate to maintain the humidity. Four plants were inoculated with each isolate. As a control, four plants were inoculated with noncolonized PDA plugs. All treated plants were kept in a greenhouse at 28 ± 3°C and 70% relative humidity. Foliar blight and necrotic lesions around inoculation points were observed about 65 days after inoculation, and 50% of inoculated trees exhibited symptoms, whereas the control trees remained symptomless. Neoscytalidium dimidiatum was successfully reisolated from symptomatic tissue via morphological analysis. To our knowledge, this is the first report of root collar canker caused by N. dimidiatum on J. mimosifolia. Neoscytalidium dimidiatum has a wide range of hosts, including pitaya, pine, mulberry, pear, grape, locust tree (Luo et al. 2024). This finding will help in controlling of the disease epidemic.

CT hyperdense lesions after endovascular therapy in acute ischemic stroke: imaging findings and clinical significance.

Background The hyperdense lesion on non-contrast CT (NCCT) is a common postoperative phenomenon in acute ischemic stroke (AIS) patients who are treated with endovascular therapy (EVT). Both contrast extravasation and hemorrhagic transformation presented hyperdense lesions on NCCT, which are sometimes difficult to distinguish them. Summary of Review Radiographic findings are important for identifying contrast extravasation and hemorrhagic transformation. We recommended a standardized follow-up protocol involving imaging and clinical evaluation as it will allow neurologists and neuroradiologists to reveal the relationships between these hyperdensities and various clinical outcomes. Key Messages Dual-energy CT and susceptibility weighted imaging are capable of distinguishing contrast extravasation and hemorrhagic transformation at an early stage after EVT. However, in institutions without access to such technology, a follow-up protocol based on NCCT is crucial.