Nontuberculous mycobacterial pulmonary disease (NTM-PD) is a chronic and challenging infectious condition with rising global prevalence, and its bronchiectatic subtype is particularly difficult to treat with conventional regimens due to frequent adverse effects, thus necessitating novel therapeutic approaches. This report presents a 42-year-old female patient diagnosed with bronchiectatic NTM-PD, characterized by persistent cough, sputum production, and hemoptysis, which was confirmed through radiological findings and microbiological testing. Following intolerance to conventional antimicrobial therapy, the patient was treated exclusively with nebulized inhalation of human umbilical cord mesenchymal stem cell-derived exosomes (HucMSC-Exos). This intervention led to significant alleviation of respiratory symptoms, stabilization of pulmonary function, and marked improvement in immunological parameters. In conclusion, nebulized HucMSC-Exos therapy demonstrated successful outcomes in this case, improving clinical symptoms, pulmonary function, and immune status, which suggests its potential as a viable and innovative therapeutic strategy for managing this complex condition and warrants further clinical investigation.

This retrospective study examines the clinical outcomes and immune reconstitution dynamics in patients with severe aplastic anemia (SAA) exhibiting mixed chimerism (MC) compared with those with full donor chimerism (FDC) following HLA-matched hematopoietic stem cell transplantation (HSCT). Analysis of propensity score-matched cohorts (23 MC vs 69 FDC) revealed comparable 5-year overall survival (OS: 87.0% vs 92.8%, P = .433) but significantly inferior failure-free survival (FFS: 47.8% vs 87.0%, P < .001) in patients with MC due to a higher incidence of graft failure (52.2% vs 8.7%, P < .001). Longitudinal immune profiling revealed delayed recovery of myeloid and lymphoid lineages in patients with MC at 12 months post-HSCT, with pronounced deficits in adaptive immunity. Specifically, CD8+CD28+ T cell counts were consistently reduced at 1 month (median, 20 vs 41 cells/μL, P = .049), 3 months (median, 86 vs 153 cells/μL, P = .024), and 6 months (median, 109 vs 160 cells/μL, P = .001), and CD4+CD25+ T cells were diminished at 6 months (median, 11 vs 20 cells/μL, P = .006). Multivariate analysis revealed that elevated CD8+CD28+ T cell levels at 3 months (≥140 cells/μL) were an independent predictor of improved FFS (HR = 0.30, P = .035). These findings highlight MC-associated immune dysregulation, particularly impaired CD28-costimulated T cell and CD4+CD25+ T cell reconstitution, as a key mediator of graft instability. This study underscores the prognostic value of early immune monitoring and suggests therapeutic strategies that target T cell recovery to mitigate MC-related risk in patients with SAA.

Mitochondrial dysfunction and dysregulated proteolysis drive Huntington's disease (HD), tauopathy, and related neurodegenerative disorders. Calpain-2, a Ca2+-activated protease restrained by calpastatin (CAST), is pathologically overactivated, yet no therapies directly target this axis. We identify A36, a brain-penetrant small molecule derived from CHIR99021 that selectively stabilizes the CAST-calpain-2 complex without inhibiting GSK3. A36 acts as a protein-protein interaction stabilizer, enhancing CAST-calpain-2 binding, preventing CAST degradation, and thereby limiting calpain-2 activation and mitochondrial damage. In patients with HD induced pluripotent stem cell-derived neurons and mutant mouse striatal neurons, A36 normalized mitochondrial morphology and membrane potential, reduced oxidative stress, and improved survival. In vivo, A36 displayed favorable pharmacokinetics and central nervous system exposure; treatment reduced striatal neurodegeneration, mutant huntingtin aggregation, and motor deficits in HD R6/2 mice, and lowered phosphorylated tau, neuroinflammation, and cognitive decline in tauopathy PS19 mice. These findings establish pharmacological stabilization of CAST-calpain-2 as a therapeutic strategy and position A36 as a mechanism-selective modulator with broad neurodegenerative disease potential.

Neural progenitor cells exhibit developmental plasticity as they can commit to distinct developmental trajectories. The male-specific lethal complex (MSLc) is linked to multiple developmental disorders, suggesting a role in neural fate commitment. To dissect MSLc function, we used a multipronged approach combining chronic and acute depletion models. Knockout of the MSLc scaffolding component MSL1 caused embryonic lethality by E10.5 (embryonic day 10.5), and single-cell multiomics revealed altered cell population composition across multiple germ layer-derived lineages, including neuroectoderm. Two-dimensional directed differentiation models showed that the MSLc facilitates accessibility at regulatory elements during early stages of neurogenesis. Neurodevelopmental genes displayed reduced enhancer-promoter contacts and failed to reach appropriate expression levels when the MSLc was absent early in neural differentiation. In contrast, MSLc loss at later stages did not recapitulate this phenotype, indicating that MSLc-mediated gene priming is a key mechanism enabling timely activation of lineage-specifying transcriptional programs.

Acute myeloid leukemia (AML) is a hematopoietic malignancy caused by abnormal proliferation and differentiation of blasts. PRMT5, a methyltransferase that catalyzes symmetric dimethylation of arginine (SDMA) residues, has been implicated in cancer stem cell homeostasis and shown to be a potential therapeutic target in AML. However, given the toxicity of complete PRMT5 inhibition, there is a need to identify effective synergistic therapies. Through a targeted screen of compounds that inhibit key nodes of PRMT5-regulated pathways, we identified a synthetic lethality between inhibition of PRMT5 and LSD1, a lysine demethylase known to affect AML blast differentiation. The two inhibitors broadly reshape the transcriptome of targeted cells and synergize to promote AML differentiation and eventually growth inhibition and apoptosis, in a p53-dependent manner. To leverage this synthetic lethal interaction, we generated new dual compounds to inhibit both enzymes and recapitulated the effects of the drug combination. Our results uncover an unexpected convergence of PRMT5- and LSD1-regulated targets, paving the way for new therapeutic opportunities.

Glucocorticoid (GC)-induced osteonecrosis of the femoral head (ONFH) involves bone marrow-derived mesenchymal stem cell (BMSC) apoptosis and dysregulated osteo-adipogenic differentiation. While aberrant H19 promoter methylation and expression have been linked to various bone metabolic disorders such as osteoporosis and osteosarcoma, their specific role in the pathogenesis of GC-induced ONFH remains largely unexplored.

The PIWI-interacting RNA (piRNA) biosynthesis pathway is best studied for its role in suppressing Drosophila germline transposable elements. Piwi, the founding member of the pathway, is involved in adult intestinal stem cell (ISC) homeostasis. Whether a broader role of the PIWI pathway exists in the intestine remains unknown. Here, we characterize a role of the PIWI family protein Aubergine (Aub) in ISCs. While dispensable for basal ISC self-renewal, upregulation of Aub by damage-induced reactive oxygen species drives regenerative ISC proliferation through increased protein synthesis, including translation of ISC factors Myc and Sox21a. Unexpectedly, such roles of Aub in ISCs appear uncoupled from its piRNA regulatory function. Additionally, Aub and mammalian PIWIL1 mediate tumorigenic intestinal growth in Drosophila and human organoids, respectively. Our results reveal regulated protein translation as a fundamental aspect of regenerative ISC function and discover a central role of Aub in such process.

The adult B cell pool is a mosaic comprising short-lived naive B cells and long-lived memory. Using genetic time stamping, we have previously shown that early-life-origin (ELO) B cells contribute substantially to the adult mouse immune system. Here, we show that they share a memory-like signature, with ELO B-1 cells being enriched for the PD-L2/CD80 double-positive (DP) immunophenotype. Indeed, microbial antigen exposure in neonates expands distinct specificities within the DP B-1 cell compartment, identifying it as a reservoir of immunoglobulin (Ig)M memory. B cell chronic lymphocytic leukemia (CLL) is a disease marked by the accumulation of memory-like cells. By applying time stamping to a mouse model of unmutated CLL, we demonstrate that leukemic expansion is driven by B-1 clones that arise prior to postnatal day 10. Importantly, B-1 cells in mice and humans share molecular features with unmutated CLL, altogether supporting a potential contribution of ELO B cells to this disease.

Overcoming the limitations of synthetic vascular grafts in the development of biocompatible and regenerative vessels remains a long-term objective in tissue engineering. In this study, we engineered large-diameter fibrin-based vascular grafts with a target inner diameter of 21 mm replicating all three layers of the native human vessel wall in vitro.

Recent studies have highlighted the impact of copper-induced cell death (cuproptosis) on cancer progression, prognosis, and treatment, but it remains unclear whether cuproptosis-related genes (CRGs) play any role in the glioma tumor microenvironment (TME). The CRGs expression patterns in TCGA glioma samples were evaluated based on genetic and transcriptional alterations identifying three different molecular groupings and showing that CRGs changes were linked to clinical characteristics, prognosis, and TME infiltration. Machine learning algorithms were then used to develop an overall survival score for cuproptosis-related prognostic genes (CRPGs), and its prognostic ability was validated for glioma patients. An elevated CRPGs score indicates a heightened mutation burden, increased glioma metabolism, compromised immunity, and strong correlation with both the cancer stem cells (CSC) index and medication sensitivity to chemotherapeutics. This extensive examination of CRGs in gliomas showed their possible significance in the tumor microenvironment as well as their prognostic value. This extremely precise CRPGs nomogram has furthered our understanding of cuproptosis in gliomas, which will allow new approaches to prognosis and immunotherapy development.

Controlling the proliferation and immune evasion of cancer cells can lead to effective strategies for cancer treatment. Thymoquinone, a bioactive compound derived from Nigella sativa exhibits a potent anti-tumor activity. It can be used as a therapeutic approach because it stabilizes the G-quadruplex structure in the promoter regions of oncogenes. This study aims to investigate the effects of thymoquinone on CD55, an inhibitor of the complement system, and CD114, which is involved in cancer cell proliferation. Real-time PCR and Western blot were conducted to verify the expression of CD55 and CD114 in patients' tumor samples, HMEC cells, MCF-7 cells, and MCF7-cancer stem cells (MCF7-CSCs). MCF-7 cells were treated with thymoquinone, and their biological behavior was evaluated using proliferation, migration, and wound-healing assays. The result indicated that CD55 and CD114 were induced among the patient's samples. The same result is followed by MCF-7 cells and MCF7-CSCs. Treatment of MCF-7 and MCF7-CSCs with thymoquinone effectively downregulated CD55 and CD114 and suppressed the stemness markers Sox2 and Nanog. Promoter analysis revealed the putative G-quadruplex sequences in the CD55 and CD114 genes. Thymoquinone binds to them at the CD55 and CD114 promoters, thereby limiting mRNA expression. Additionally, the inhibition of their expression reduced cell movement and growth, as verified by biological assays. In summary, treating breast cancer cells with thymoquinone could stabilize the G-quadruplex structure on the promoter regions of CD55 and CD114 and hinder their mRNA expression. Therefore, restoring immune recognition and inhibition of proliferation. Hence, thymoquinone could be a potent target for breast cancer therapeutics.

The number of pet dogs is increasing, and the number of working dogs (e.g., guide dogs, police dogs) is also gradually increasing. Skin wounds are a common clinical problem in dogs and tend to be more common in the clinic as mechanical wounds. The healing process of skin wounds is often influenced by a variety of factors, including infection, nutritional status, and immune response, while wound healing is more difficult in dogs with diabetes or aging dogs. Mesenchymal stem cells (MSCs) play an important role in skin healing and regeneration with their multidirectional differentiation potential and immunomodulatory function. However, the application of MSCs alone for the treatment of skin wounds may have certain limitations, such as low cell survival and a lack of localization. Therefore, it is important to find methods that can enhance the therapeutic effect of MSCs. Secreted protein acidic and rich in cysteine (SPARC), an extracellular matrix protein widely involved in regulating biological processes such as cell proliferation, migration, and matrix production, may enhance the efficacy of MSCs in skin wound healing. This study aims to systematically evaluate the therapeutic efficacy of SPARC-overexpressing adipose-derived mesenchymal stem cells (ADSCs) in promoting skin wound healing by establishing wound models in normal, diabetic, and aged mice and dogs, thereby validating their potential under diverse physiological and pathological conditions. For in vitro validation, we used hydrogen peroxide (H2O2) to induce Human Umbilical Vein Endothelial Cell (HUVEC) and Human Keratinocyte Cell (HaCaT) injury. All animals were randomly assigned to six experimental groups as follows: (1) Model group: Untreated wound (negative control); (2) HY group: Hydrogel alone (vehicle control); (3) Con group: Control-ADSCs (cell control); (4) Con-Exo&HY group: Control-ADSC exosomes in hydrogel; (5) SPARC group: oe-SPARC-ADSCs (treatment); (6) SPARC-Exo&HY group: oe-SPARC-ADSC exosomes in hydrogel (treatment). Separately, HUVEC and HaCaT cells were assigned to four experimental conditions: a blank control group, a model group, a control-ADSC-treated group, and an oe-SPARC-ADSC-treated group. ADSCs modified by SPARC significantly promoted re-epithelialization integrity, collagen deposition, inflammation reduction, angiogenesis, and hair follicle regeneration during wound healing in dog skin. HUVEC and HaCaT cells proliferated after adding oe-SPARC-ADSCs cell supernatant. Meanwhile, quantitative proteomic sequencing data analysis showed that SPARC could promote skin wound healing by enhancing cell adhesion, hyaluronic acid binding, and vascular smooth muscle contraction of ADSCs. Both in vitro cellular assays and in vivo wound-healing models suggest that the combination of SPARC and ADSCs for the treatment of skin wounds has broad application prospects.

Intervertebral disk pathology, including disk herniation and degeneration, is a major contributor to chronic low back pain, and when conservative treatment fails, surgical management often involves discectomy-based procedures that leave residual annulus fibrosus (AF) defects associated with reherniation and progressive degeneration. These limitations have motivated interest in regenerative strategies using biomaterial scaffolds; however, reproducing the hierarchical, angle-ply architecture of the AF remains challenging. Here, we present a single-step extrusion-based 3D-printing approach to fabricate polycaprolactone (PCL) scaffolds with aligned microscale surface grooves that promote AF-like organization. Patterned nozzles with circumferential peaks generated uniaxial concave microgrooves (10-17 µm wide) directly during printing, enabling formation of multilamellar angle-ply constructs. Human bone marrow-derived mesenchymal stem cells cultured on patterned scaffolds aligned longitudinally within concave grooves, forming end-to-end arrays that guided extracellular matrix deposition. Gene expression analysis showed that topographical cues governed cellular organization without significantly altering gene expression profiles, while TGF-β3 supplementation upregulated outer AF-associated markers, including COL1, COL12, SFRP2, MKX, MCAM, and SCX. TAGLN expression increased specifically on patterned scaffolds in the absence of TGF-β3, indicating an association between microgroove-guided cellular organization and TAGLN expression, warranting further investigation into potential tension-related mechanisms. This novel single-step extrusion-printing approach leverages custom nozzle geometry to impart concave microgrooves, facilitating scalable fabrication of multilamellar angle-ply scaffolds that induce aligned cellular organization and support potential applications in annulus fibrosus repair, as well as mechanobiological studies of anisotropic musculoskeletal tissues.

Stem cell heterogeneity represents a critical yet underexplored variable in laser-assisted regenerative strategies. While photobiomodulation has been shown to influence mesenchymal stem cell (MSC) behavior, it remains unclear whether stem cell maturation status modulates responsiveness to Er,Cr:YSGG irradiation. This study investigated the differential response of magnetically separated STRO-1+ and STRO-1- SHED subpopulations to low-power Er,Cr:YSGG laser stimulation (0.10 W and 0.25 W), focusing on viability, extracellular matrix production, and lineage commitment. STRO-1+ cells comprised 13.4% ± 1.2% of the total Stem Cells from Human Exfoliated Deciduous teeth (SHED) population. Laser exposure did not impair metabolic activity in either subpopulation. Collagen synthesis demonstrated a power- and time-dependent increase, with maximal enhancement observed in STRO-1+ cells at 0.25 W after 7 days. Laser irradiation selectively promoted osteogenic differentiation, as evidenced by increased alkaline phosphatase (ALP) expression at 0.10 W and enhanced mineral deposition, while chondrogenic potential remained unaffected and adipogenesis was reduced following 0.10 W exposure. These findings suggest that ALP expression is temporally and power-dependently modulated during osteogenic progression. Overall, Er,Cr:YSGG photobiomodulation does not uniformly affect heterogeneous SHED populations but modulates lineage allocation and extracellular matrix deposition in a maturation- and power-dependent manner. Integrating stem cell subpopulation selection with laser-based bioactivation may represent a strategy to refine regenerative endodontic and biomaterial-guided therapies.

Zirconium is a widely used material in the field of dentistry, employed for implants and their components as well as for the creation of crowns and veneers. Given that its biocompatibility has been studied and demonstrated in various fields of application, it is necessary to analyze how surface modification of this material influences its properties. The purpose of this study was to analyze the biocompatibility, initial adhesion (48 h), and morphology of periodontal ligament stem cells (PDLSCs) seeded on different zirconium surfaces treated with laser micropatterning, as well as plastic coverslips as a control. The Neubauer chamber was used to count the cells adhered to each of the sets, and confocal and scanning electron microscopy were employed to examine the adhesion and morphology of periodontal ligament stem cells on each of the zirconium surfaces studied. Results: Statistically significant differences were found in terms of primary cell adhesion, with sets 3 (grid topography) and 4 (channel topography) showing the most favorable characteristics for fibroblast adhesion. It was concluded that regular and moderately rough surfaces promoted better cell proliferation and development.

Achilles tendon rupture often leads to poor functional recovery due to limited self-healing, with mitochondrial dysfunction in tendon stromal cells (TSCs) being a key factor in disease progression. Here, we developed adipose-derived stromal cell (ADSC) membrane-coated mitochondria (Mito-NPs) to target this dysfunction and evaluate their therapeutic potential for tendon repair. Mito-NPs exhibited uniform size, stable surface charge, and effective membrane coating. In lipopolysaccharide-induced inflammatory TSCs, Mito-NPs enhanced oxidative phosphorylation, improved mitochondrial metabolic homeostasis, and reshaped gene expression profiles to normalize TSC functional phenotypes, including inflammation, migration, and collagen synthesis. When encapsulated in a reactive oxygen species (ROS)-responsive hydrogel (Mito-NPs@HG) and implanted into rat Achilles tendon injuries, Mito-NPs@HG improved gait function, decreased local inflammation, and promoted histological repair of damaged tendons by enhancing collagen organization and reducing inflammation. Our findings demonstrate that ADSC membrane-coated mitochondria effectively rescue TSC dysfunction and facilitate tendon regeneration, providing a promising translational strategy for treating tendon injuries.

Acute myeloid leukemia (AML) is a molecularly heterogeneous neoplasm of hematopoietic stem and progenitor cells. The advent of high-resolution genomic sequencing has uncovered several genetic drivers of AML which spurred a surge of therapies that target the disease at a mutational, clonal, or epigenetic level. Currently, the molecular profiling of AML patients before treatment is commonplace and crucial for ensuring that patients receive the most optimal therapy for any driver mutations they may have. Here, we detail the current targeted therapies available for AML: specifically, those targeting the BCL2 family (venetoclax), FLT3 (midostaurin, gilteritinib, quizartinib), IDH1/2 (enasidenib, ivosidenib), and MENIN (revumenib, ziftomenib). In addition, we outline potential mechanisms of resistance against these therapies, as well as efforts being taken to prevent or bypass them.

Malignant melanoma is skin cancer arising from genetically altered melanocytes. Recently, a complex relationship between melanoma and chronic inflammation has been highlighted, representing an excellent condition for tumor development. Microalgae have been shown to be a promising source of bioactive compounds for drug discovery. In this study, we investigated Halamphora sp. (BEA0050) to identify possible compounds with immunomodulatory activity. The most active fraction (fraction D) showed anti-inflammatory activity against human melanoma cancer cells (A2058) stimulated using lipopolysaccharide (LPS) to induce an inflammatory phenotype. Chemical profiling of the bioactive fraction using chromatography and high-resolution mass spectrometry (UHPLC-HR-MS) revealed hydroxypheophorbide a, a breakdown product of chlorophyll a. In order to investigate the mechanism of action, the TNF-α release was detected through ELISA sandwich assays in A2058 cells and through confocal microscopy in LPS-stimulated HaCaT cells. Gene expression of principal pro-inflammatory cytokines and pathways was detected through real-time PCR, which showed the down-regulation of the inflammatory pathway in LPS-induced A2058 and HaCaT cells treated with 12.5 µg/mL of fraction D. This study reports for the first time the anti-melanoma and anti-inflammatory activities of Halamphora sp., identifying protein mediators and highlighting its biotechnological potential.

Diagnostic evaluation and management of nontraumatic osteonecrosis of the femoral head (ONFH) vary substantially. This systematic review was conducted to inform development of the Association Research Circulation Osseous (ARCO) clinical practice guideline for diagnosis and treatment of ARCO stages I to III ONFH.

The development of inks with suitable rheological, physicochemical, mechanical, and biological properties is crucial for the successful fabrication of functional scaffolds via extrusion-based 3D printing. In this study, collagen/chitosan hydrogels with varying polymer ratios were developed and characterized to evaluate their printability and suitability for cartilage tissue engineering. Rheological analyses revealed that all samples exhibited shear-thinning behavior and solid-like viscoelasticity, with the formulation of an 80:20 COL/CHI ratio (20CHI) demonstrating optimal filament formation and dimensional stability. Physicochemical analyses confirmed the preservation of the collagen triple helix and the formation of hydrogen bonding between chitosan and collagen. 20CHI scaffolds showed swelling capacity and high cohesiveness. In vitro studies confirmed the cytocompatibility of the scaffolds with murine fibroblasts and the ability of the scaffolds to promote adhesion, proliferation, and extracellular matrix production of both chondrocytes and adipogenic mesenchymal stem cells (aMSCs). Quantification of sulfated glycosaminoglycan (sGAG) indicated sustained matrix deposition over 28 days, particularly by chondrocytes. These findings demonstrate that 20CHI hydrogel is a promising candidate for 3D printing of biomimetic scaffolds for cartilage regeneration.