Motor neurons in the brain and spinal cord begin to die off in Amyotrophic lateral sclerosis (ALS), a disease that can be fatal. Molecular pathways in neurological disease, especially ALS, remain a challenge in the medical sciences. In this disease, a disorder in both astrocytes and oligodendrocytes can cause the disease to progress. This study aimed to investigate the molecular mechanisms and find key elements between these two cells in ALS with a bioinformatics perspective. In this study, using integrated and continuous bioinformatics analytics by various tools and databases, we investigated genes, protein products, and miRNAs between astrocytes and oligodendrocytes. The obtained data were involved in the Cellular senescence, actin cytoskeleton, and cell cycle signaling pathways. Then, after careful evaluation of the information, TP53, MDM2, KRAS, PTPRC, and GSK proteins were candidates, which are regulated by hsa-miR-564, hsa-miR-496-5p, hsa-miR-324-5p, hsa-miR-296-5p, and hsa-miR-4258-3p miRNAs. Finally, the four genes had a more robust and better relationship in this study between astrocyte and oligodendrocyte-derived ALS.

N6-methyladenosine RNA methylation, an essential post-transcriptional modification, dynamically regulates RNA metabolism and plays a crucial role in neuronal function. Growing evidence suggests that dysregulated N6-methyladenosine modification contributes to the pathogenesis of neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, multiple sclerosis, and amyotrophic lateral sclerosis. However, the precise mechanisms by which N6-methyladenosine modification influences these conditions remain unclear. This review summarizes the role of m6A modification and its associated regulators in neurodegeneration, focusing on their involvement in key pathological processes. In Alzheimer's disease, m6A modification contributes to synaptic dysfunction, mitochondrial damage, and neuronal apoptosis. Evidence from APP/PS1, 5XFAD, tau transgenic, and Drosophila models demonstrates that regulators such as METTL3 and FTO influence Alzheimer's disease progression through neuroinflammation, circRNA dysregulation, and autophagy-related mechanisms. In Parkinson's disease, altered N6-methyladenosine regulator expression affects dopaminergic neuron survival and stress responses by modulating mRNA stability and autophagy-related lncRNAs. In multiple sclerosis and amyotrophic lateral sclerosis, N6-methyladenosine affects immune activation, myelin repair, and the regulation of disease-associated genes such as TDP- 43. Beyond N6-methyladenosine, other RNA methylation modifications-such as m1A, m5C, m7G, uracil, and pseudouridine-are implicated in neurodegenerative diseases through their regulation of mitochondrial function, RNA metabolism, and neuronal stress responses. Additionally, N6- methyladenosine exhibits cell type-specific functions: in microglia, it regulates inflammatory activation and phagocytic function; in astrocytes, it modulates metabolic homeostasis and glutamate-associated neurotoxicity; in neurons, it affects synaptic function and neurodegeneration-related gene expression; and in adult neural stem cells, it controls differentiation, neurogenesis, and cognitive plasticity. Recently, several small-molecule inhibitors targeting METTL3 or FTO have been developed to modulate N6-methyladenosine modification, providing new opportunities for disease intervention, with the targeting of N6-methyladenosine-related pathways emerging as a promising therapeutic strategy. However, challenges persist in optimizing the specificity and delivery of these therapeutic approaches.

Acute graft-versus-host disease (aGVHD) remains a life-threatening complication that limits the efficacy and application of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Early identification and diagnosis of aGVHD is significant for improving the outcomes of allo-HSCT. We retrospectively analyzed the dendritic cell (DC) subsets in peripheral blood (PB) of patients who received allo-HSCT before transplantation and at engraftment to evaluate the correlation between DC subsets and the occurrence of aGVHD. The results showed that the proportion and count of myeloid dendritic cells (mDCs) before transplantation were positively correlated with the occurrence and severity of aGVHD, while the proportion and count of mDCs at engraftment were negatively correlated with those. In addition, patients with higher proportion and count of mDCs before transplantation developed aGVHD earlier, whereas those without aGVHD had the lowest proportion and count of mDCs before transplantation. The proportions and counts of DC subsets at engraftment were not significantly correlated with the onset time of aGVHD. Receiver operating characteristic (ROC) curve analysis demonstrated that the count of mDCs in PB before transplantation had greater sensitivity and specificity in predicting aGVHD (AUC = 0.8002, P < 0.0001) compared to plasmacytoid DCs (pDCs) and total DCs. Multivariate logistic regression analysis confirmed that the count of mDCs in PB before transplantation and at engraftment were independent factors in predicting the occurrence of aGVHD (OR = 9.907, P = 0.018, 95% CI 1.473-66.639; OR = 0.059, P = 0.023, 95% CI 0.005-0.673). Therefore, mDCs in PB play an important role in the development of aGVHD and may serve as a promising predictor of aGVHD for patients undergoing allo-HSCT.

Enhanced tendon‒bone healing is of critically importance for achieving optimal postoperative recovery following a rotator cuff tendon tear (rotator cuff tears, RCTs). Although RCTs patch-augmented scaffolds demonstrate clinical potential, there is a paucity of reports on biodegradable scaffolds that effectively integrate high strength and bioactivity. Inspired by the composition and aligned nanofibrous structure of the natural fish bladder matrix (fish swim bladder, FSB), we employed a gallium (Ga)‒tannic acid (TA) metal‒polyphenol network (MPN)-modified decellularized fish bladder matrix (GaPP@FSB) as a novel biomaterial to address this problem. Ga-TA MPN represents a "two birds with one stone" modification strategy that allows GaPP@FSB to demonstrate commendable mechanical strength alongside multiple biological activities, including antibacterial, antioxidant, anti-inflammatory and osteogenic differentiation promotion. Furthermore, GaPP@FSB regulates the focal adhesion-based mechanical signal transduction pathway in tendon stem/progenitor cells (TSPCs), thereby activating the α5β1/Akt/PI3K pathway to induce tenogenic differentiation. Additionally, this scaffold exhibits remarkable anti-inflammatory and antibacterial activities. In a rat RCTs model, GaPP@FSB promoted regeneration at the tendon‒bone interface while restoring both rotator cuff biomechanics and joint movement function. Consequently, this biomaterial derived from natural FSB has outstanding biosafety and biological activity, making it a highly promising candidate for clinical applications in both tendon repair and the restoration of the tendon‒bone interface.

Over the past decade, there has been no clear evidence regarding the comparative effectiveness of bone-marrow mononuclear cell (BMMNC) therapy in patients with chronic heart failure (HF).

This abstract has been removed: please see Elsevier policy on article withdrawal (https://www.elsevier.com/about/policies-and-standards/article-withdrawal). This article has been removed at the request of the author. This abstract has been removed because it was not presented at the 2025 BMT Tandem Meeting.

This abstract has been removed: please see Elsevier policy on article withdrawal (https://www.elsevier.com/about/policies-and-standards/article-withdrawal). This article has been removed at the request of the author. This abstract has been removed because it was not presented at the 2025 BMT Tandem Meeting.

Cardiac tissue macrophages are crucial components of the immune system and tissue homeostasis. Traditionally, these macrophages have been classified into three primary lineages: yolk sac blood island-derived erythromyeloid progenitor (EMP), yolk sac hemogenic endothelial-derived late-EMP, and hematopoietic stem cell (HSC)-derived macrophages. These classifications have shaped our understanding of the developmental origin of macrophages in the heart. However, recent studies have significantly shifted this perspective by revealing that the heart itself possesses an intrinsic source of macrophages, independent of the traditionally recognized hematopoietic sources. This discovery has added a new dimension to our understanding of macrophage biology in the context of cardiac development. Our recent work has provided compelling evidence that endocardial cells exhibit hematopoietic potential during embryonic day (E)8.5 to E10. This discovery challenges the previously held belief that macrophages in the heart are exclusively derived from EMP or HSC. Endocardial cells give rise to a distinct population of cardiac tissue macrophages that play vital roles in heart morphogenesis. These findings open up new avenues for understanding how macrophages contribute to heart formation, homeostasis, and their disruption. This review summarizes the latest findings on the role of endocardial-derived macrophages, along with other macrophage lineages, in contributing to heart development and the maintenance of cardiac homeostasis.

Hemocytes represent a pivotal element of insect defense against pathogens through the mechanisms of cellular and humoral immunity. However, hemocyte types and functions exhibit variation across different insect taxa, leading to the lack of a standardized classification system for insect hemocytes. To gain insight into the immune mechanisms of the house fly Musca domestica, we used a combination of morphological observations and single-cell RNA sequencing techniques (scRNA-seq) to taxonomically characterize house fly larval hemocytes and analyze their immune function. As a result, five different types of hemocytes were identified from house fly larvae, granulocytes (GR), spherulocytes (SP), plasmatocytes (PL), prohemocytes (PR), and oenocytoids (OE). On the basis of microscopic observations, flow cytometry analysis and gene expression profiles, the immune functions of house fly hemocytes were hypothesized to be as follows: the GRs are responsible for phagocytosis, the SPs are highly expressive of lectins, and the PLs are highly expressive of antimicrobial peptides (AMPs) and are involved in nodule formation, the PRs act as progenitor cells and retain the differentiation potential of stem cells, and the OEs are involved in melanization reactions mainly through the expression of phenoloxidase (PO). Based on the scRNA-seq data, the marker genes for each type of hemocyte were identified. The present study unveils the heterogeneity of house fly hemocytes in terms of morphology, gene expression characteristics, and function, thereby establishing the foundation for an in-depth understanding of the immune mechanisms in house fly.

The doublesex (dsx) gene has a conserved role in sex determination in insects, controlling sexual development and mating behavior. Although dsx is known to participate in these critical functions, its role in insect sex determination remains not fully elucidated. Given the economic importance of the rice pest Chilo suppressalis, we employed this species as a model to investigate the function of its dsx homolog. We cloned and characterized the Csdsx gene, which is 1,123 bp in length and encodes four sex-specific proteins: three female-specific isoforms of 252, 258, and 254 amino acids, respectively, and one male-specific isoform of 290 amino acids. Phylogenetic analysis revealed that Csdsx is highly conserved within Lepidoptera, containing two domains: DM DNA binding domain and DSX dimer domain. Analysis of transcripts produced from a mini-dsx construct transfected into human HEK293T cells indicates that the female-splicing pattern is the default mode. Quantitative real-time PCR showed that among various developmental stages, Csdsx expression peaked at the first instar larval stage and showed tissue-specific, stage-dependent patterns, with notably high levels in the larval midgut, pupal fat body, and adult thorax in both sexes. In comparison to wild-type (WT) adults, Csdsx-knockout individuals exhibited malformations in their external genitalia, and female wing patterns became masculinized. Dissections revealed that knockout females had a reduced number of mature oocytes, while knockout males displayed a smaller testis area. Furthermore, when Csdsx-knockout females were paired with wild-type males, their mating behavior was significantly impaired. RNA-seq revealed that Csdsx disruption led to sex-biased gene expression shifts, including upregulation of male-associated genes (e.g., OBPs, PBPs, trypsin) and downregulation of female-specific genes (vitellogenin, FAS), indicating partial masculinization at the transcriptional level. These findings underscore the critical role of the dsx gene in reproductive development and sexual dimorphism in C. suppressalis.

Cancer cachexia is a multifaceted metabolic syndrome characterized by severe loss of skeletal muscle and adipose tissue, diminishing both quality of life and survival in cancer patients. Despite its prevalence, effective treatments for cancer cachexia remain limited. The JAK/STAT signaling pathway has been identified as a key driver of muscle atrophy in cachexia.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most prevalent liver disease worldwide, primarily caused by poor lifestyle choices and dietary habits. Caulis Bambusae In Taenias, commonly known as Zhuru in Chinese, is a traditional Chinese medicinal material from the Bambusoideae family, frequently used for liver detoxification, liver protection, and lowering lipids.

Breast cancer (BC) is one of the most prevalent malignant tumors among women, with estrogen receptor (ER)-positive patients constituting approximately 70% of all cases. Endocrine therapy is currently a treatment option for patients with ER-positive BC; however, the development of resistance significantly limits the effectiveness of this treatment. Nano-curcumin (Nano-CUR) possesses anticancer properties and enhances bioavailability by improving the hydrophobic character of curcumin (CUR). However, the underlying mechanism by which Nano-CUR affects tamoxifen (TAM) resistance in ER-positive BC remains unknown. Here, we found that Nano-CUR promoted apoptosis and cell cycle arrest, inhibited cell proliferation and reduced the levels of cancer stem cells (CSCs)-related markers, including octamer-binding protein (OCT4), Nanog homeobox (NANOG) and sex-determining region Y-box 2 (SOX2) in TAM-resistant BC cells. Additionally, Nano-CUR demonstrated the ability to inhibit tumor malignant progression in TAM-treated BC mice. Mechanistically, Nano-CUR blocked the activation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway in MCF-7/TAM and T47D/TAM cells. The activation of this pathway by its activators (PI3K activator 740Y-P, AKT activator SC-79, and mTOR activator MHY1485) effectively alleviated the anti-tumor effect induced by Nano-CUR in TAM-resistant BC cells. Collectively, these findings reveal that Nano-CUR contributes to the reduction of tumorigenesis and TAM resistance in ER-positive BC cells by inhibiting the PI3K/AKT/mTOR signaling pathway.

T cells play pivotal roles in orchestrating immune responses after allogeneic hematopoietic stem cell transplantation. Donor T cells facilitate the reconstitution of the recipient's immune system and Enhancer of Zeste Homolog 2 (EZH2) is crucial for T-cell differentiation and plasticity. Inhibitors of EZH2 may protect patients undergoing allogeneic stem cell transplantation from developing graft-versus-host disease without compromising the beneficial graft-versus-leukemia effect. Here, we investigated the impact of SHR2554, a novel and highly selective EZH2 inhibitor, on the subset ratios and polyfunctional responses of peripheral T-cells from hematopoietic stem cell donors METHODS: Cell proliferation, cell cycle, and apoptosis were analyzed using Cell Counting Kit-8 assays and flow cytometry. Western blotting was employed to detect the abundance of related proteins. RNA-sequencing was used to analyze gene expression after SHR2554 treatment RESULTS: SHR2554 inhibited proliferation, promoted apoptosis, and induced cell cycle arrest in donor T cells. SHR2554 also preserved CD4+/CD8+ T cell ratios while selectively depleting naïve (TN) and terminally differentiated effector (TEff) subsets and concurrently expanding central memory (TCM), effector memory (TEM), and regulatory T cell (Treg) subsets across CD3+/CD4+/CD8+ populations. SHR2554 significantly attenuated CD107a expression and IFNγ/TNFα secretion from CD3+/CD4+/CD8+ T cells, thereby suppressing polyfunctional responses across T cell subsets CONCLUSIONS: SHR2554, a potent and highly selective small-molecule inhibitor of EZH2, demonstrated a remarkable anti-proliferative effect on donor peripheral blood T cells, and modified T cell ratios and suppressed their polyfunctional responses. SHR2554 therefore has potential for treating graft-versus-host disease.

Despite their significant potential, hepatocytes derived from human embryonic stem cells (hESCs) exhibits characteristics of immature hepatocytes, similar to fetal liver cells. The quantity of the mitochondria, which functions as the main energy generators in mammalian cells, display the variation across distinct cell types with specific energy demands. However, the specific correlation between this variability and hepatocyte differentiation remains elusive. Recently, we developed a 3D suspension differentiation method to differentiate hESCs into hepatocyte spheres, and observed that resveratrol (RES) treatment further promoted hepatocyte maturation and improved the polarization of hepatocytes during hepatocyte differentiation of hESCs. We also found that the treatment with RES resulted in a significant augmentation in intracellular ATP content, mitochondrial DNA copy number, and protein subunits associated with mitochondrial biogenesis, indicating that mitochondrial function was enhanced. In addition, PGC-1α, a crucial regulator of mitochondrial biogenesis, was up-regulated during RES treatment, and eventually our further investigation revealed that RES effectively promoted hESC-derived hepatocyte maturation and polarization by enhancing mitochondrial function through PGC-1α-mediated activation of PPARγ pathway. Thus, our results demonstrated that RES-mediated enhancement of mitochondrial biogenesis could promote hepatocyte differentiation from hESCs. Therefore, this finding provides a new avenue to generate more mature hepatocytes for pharmaceutical and toxicological studies as well as for cell-based therapies.

The difficulty in regenerating osteochondral defects lies in the reduced proliferation and premature hypertrophy of chondrocytes. By maintaining the phenotype of immature chondrocytes within the superficial layer of regenerated cartilage and replicating natural cartilage microstructure, the proliferation capacity of chondrocytes can be enhanced, ensuring a continuous cell supply for cartilage repair. Therefore, utilizing biomimetic scaffolds with spatial microenvironment specificity to simulate the natural layered microstructure of osteochondral tissue is a key focus. We have designed a three-layer porous microgel scaffold with spatiotemporal controlled release of bioactive factors. In vitro, this scaffold can induce directed differentiation of rat bone marrow mesenchymal stem cells (rBMSCs) and maintain chondrocyte phenotypes at various maturation stages. In an animal model of osteochondral defects, this scaffold promoted the repair of the layered microstructure of articular cartilage. We speculate that the regenerative effect of the three-layer hydrogel system may be attributed to the precisely directed differentiation of chondrocytes and the accurate distribution of the corresponding extracellular matrix (ECM), as revealed by mRNA transcriptome sequencing. This work provides insights into chondrogenesis controlled by the microenvironment and may offer strategies for reconstructing the layered microstructure of articular cartilage.

Myeloid innate immune cells, including macrophages, neutrophils, myeloid-derived suppressor cells, and dendritic cells, represent major components of the tumor microenvironment (TME), exhibiting remarkable plasticity and dual roles in cancer progression and immune regulation. In recent years, microbial-induced innate immune memory, also termed "trained immunity" (TRIM), has emerged as a novel strategy to reprogram myeloid cells into an immunostimulatory, antitumor state. In this review, we explore the intricate landscape of myeloid cells in cancer and examine how microbial ligands, such as the Bacillus Calmette-Guérin vaccine and β-glucan, reprogram both bone marrow progenitors and tissue-resident myeloid cells to enhance inflammatory and antitumor responses. Notable findings include the hematopoietic stem and progenitor cell reprogramming by Bacillus Calmette-Guérin for sustained anticancer immunity, and the enhanced granulopoiesis and neutrophil-mediated tumor killing mediated by β-glucan-induced TRIM. These mechanisms synergize with immunotherapies, such as immune checkpoint inhibitors, by reshaping the immunosuppressive TME and enhancing adaptive immunity. However, challenges remain, including the structural complexity of microbial products, the lack of predictive biomarkers, and the need for optimized dosing and delivery strategies. Addressing these gaps by introducing precise characterization of microbial-derived ligands, biomarker-driven patient selection through large-scale clinical trials, as well as the development of novel approaches for targeted therapy will be essential to harness the full potential of microbial-induced TRIM, ultimately paving the way for more effective and durable cancer immunotherapies. SIGNIFICANCE STATEMENT: Tumor-promoting myeloid cells within the tumor microenvironment remain a major barrier to effective cancer immunotherapy. Microbial-induced trained immunity offers a novel strategy to reprogram myeloid cells into an antitumor state. This review provides a comprehensive overview of myeloid cell populations in cancer and the mechanisms underlying microbial-induced trained immunity. It also highlights preclinical and clinical evidence demonstrating the efficacy of microbial-based strategies in overcoming immunosuppression and synergizing with existing immunotherapies, offering a promising approach to improve cancer treatment outcomes.

The gut-bone axis is critical for body homeostasis, but bone loss often complicates inflammatory bowel disease (IBD) with unclear mechanisms. Here, we found that IBD mice showed reduced bone formation, with bone marrow mesenchymal stromal cells (BMSCs) favoring adipogenesis over osteogenesis. Altered N6-methyladenosine (m6A) modifications of BMSCs were confirmed in IBD mice, and further investigation revealed that the SUMOylation of FTO was involved. Nude mice with FTO-K216/357/365R mutation exhibited increased bone sizes and volumes versus control mice. Tocilizumab, an interleukin (IL)-6R monoclonal antibody, combined with AAV-FTO-3KR, mitigated bone loss and enhanced bone formation in IBD mice. Our findings reveal that SUMOylation of FTO is involved in the differentiation of BMSCs in mice with IBDs. The outcome could be blocked by redirecting differentiation toward osteoblast treatment with AAV-FTO-3KR and the clinical-stage inhibitor, tocilizumab.

The generation of loss-of-function alleles in human pluripotent stem cells (hPSCs) is used to interrogate gene function and validate reagents; however, identifying clones harboring true loss-of-function alleles remains inefficient. To address this, we present BOLT (barcoded oligos for loss-of-function targeting), a streamlined protocol that simplifies the screening process, facilitating rapid validation of loss-of-function mutations. We describe steps for designing editing tools, nucleofection, and clonal density plating. We then detail procedures for bridging PCR, isolating clones derived from a single hPSC, single-clone screening, and Sanger barcode detection. For complete details on the use and execution of this protocol, please refer to Matera et al.1.

Acute myeloid leukemia (AML) is a heterogeneous malignancy specified by clonal proliferation of hematopoietic stem cells. This study identifies novel therapeutics for AML by integrating differential gene expression (DEG) and survival analyses. Publicly available GEO microarray datasets were analyzed, including data from 615 AML patients and 22 healthy controls. Multivariate Cox regression identified hazardous genes impacting survival. Protein-protein interaction networks using CytoScape revealed hub genes such as CCT5, ZBTB16, APP, and PTPN6. Functional enrichment revealed key AML-related pathways, such as PI3K/Akt and NF-kappaB signaling. Drug repurposing using the LINCS L1000CDS2 database highlighted potential therapeutics, including 16-Hydroxytriptolide and Tryptosthin AG-1478, with roles in reversing hazardous gene expression patterns. Additional candidates such as Vemurafenib, Parthenolide and Wortmannin, demonstrated promise as targeted agents. These findings underscore the potential of integrating bioinformatics and drug discovery to identify precision medicine in AML. Further studies are warranted to validate these targets and explore their clinical utility.