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Allogeneic hematopoietic stem cell transplantation is an established curative therapy for many hematological diseases, but graft-versus-host disease remains a major cause of morbidity and mortality. Tacrolimus, a calcineurin inhibitor, is widely used for prophylaxis because it suppresses T-cell activation. However, its clinical use is complicated by a narrow therapeutic window and marked pharmacokinetic variability. Therapeutic drug monitoring based on trough whole-blood concentrations is routinely used to guide dosing, but this approach has limitations, particularly in transplantation recipients who experience rapid physiological and hematological changes. This review summarizes recent insights into determinants of tacrolimus pharmacology in hematopoietic stem cell transplantation and discusses emerging perspectives for individualized dosing. Tacrolimus exerts its immunosuppressive effects by forming a complex with FK506-binding proteins that inhibits calcineurin and suppresses activation of nuclear factor of activated T cells. Beyond this canonical mechanism, interactions with FK506-binding proteins influence the distribution of tacrolimus within blood cells. Because tacrolimus strongly divides into erythrocytes and leukocytes, whole-blood concentrations reflect systemic exposure and drug binding within circulating blood components. In recipients of hematopoietic stem cell transplantation, marked fluctuations in blood cell counts during conditioning therapy and hematopoietic recovery can alter this distribution, potentially causing changes in concentrations without corresponding changes in pharmacologically active exposure. Genetic variation in drug-metabolizing enzymes further contributes to variability in tacrolimus pharmacokinetics. In particular, polymorphisms in the gene encoding cytochrome P450 3A5 influence tacrolimus metabolism and may affect early dose requirements during the post-transplant period. Additionally, temporal fluctuations in tacrolimus exposure within individual patients are increasingly recognized as clinically relevant. Measures that capture the proportion of time during which concentrations remain within the therapeutic range provide a useful indicator of exposure stability. Tacrolimus therapy after hematopoietic stem cell transplantation is influenced by molecular pharmacology, blood cell-dependent distribution, genetic determinants of metabolism, and temporal variability in drug exposure. Integrating these factors may improve understanding of therapeutic drug monitoring and promote more individualized strategies to maintain stable immunosuppression and improve transplant outcomes.

Lung cancer is one of the most common malignancies and the leading cause of cancer-related mortality worldwide, posing a major public health challenge. Flavonoids, a large and diverse group of plant metabolites, exhibit various anticancer properties, making them promising candidates for therapeutic applications. This study evaluated the anticancer efficacy of methoxy flavonoids and elucidated their underlying mechanisms of action in A549 lung cancer cells. A549 cells were treated with various flavonoids (AKC1-AKC5), and their effects were analyzed using an MTT assay, DAPI staining, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) production, colony formation, and wound scratch tests. Molecular docking was also performed to confirm the binding of AKC1 and AKC3 to EGFR, BCL-2, and CDK-2 proteins. AKC1 and AKC3 prevented the growth of A549 lung cancer cells with IC50 of 64.57 and 19.80 μM among 5 methoxy flavonoids. AKC1 and AKC3 triggered notable alterations in the shape and reduced the colony-forming potential of A549 cells. The DAPI staining experiment demonstrated that AKC1 and AKC3 impede the growth of cancer cells through activation of apoptotic cell death. Moreover, the anticancer properties of AKC1 and AKC3 were attributed to significant inhibition of MMP and a notable ROS enhancement in a dose-related pattern. The wound scratch assay demonstrated that AKC1 and AKC3 suppressed A549 lung cancer cell migration, suggesting their anti-metastatic properties. Molecular docking studies confirmed that AKC-1 and AKC-3 bind strongly to EGFR, BCL-2, and CDK2, suggesting a multi-target mechanism that underlies their anti-proliferative and pro-apoptotic effects in A549 cells. AKC1 and AKC3 exhibited significant anticancer activity against A549 cells and may serve as promising therapeutic drugs for lung cancer treatment.

This study evaluated the polyphenol content of leaf extracts from Artemisia monosperma (AM) and investigated their antioxidant properties, cytotoxic effects, and potential to induce DNA damage in human cancer cell lines. High-performance liquid chromatography (HPLC) quantified polyphenols in methanolic (AMM), ethanolic (AME), and aqueous (AMA) extracts, identifying 13 compounds in AME and 12 in AMA. AMM exhibited the strongest antioxidant activity (IC50 = 24 µg/ml). Both AME and AMM demonstrated potent anticancer activity against HCT-116 (IC₅₀ = 0.38 µg/mL for AMM) and HUH-7 (IC₅₀ = 21.95 µg/mL for AMM) cells, while exhibiting minimal cytotoxicity toward normal skin fibroblast cells (BJ-1; IC₅₀ = 13.05 µg/mL for AMM), with AMM demonstrating particular selectivity for HCT-116 cells. AMM induced DNA fragmentation and modulated apoptosis-related gene expression (Bax, Bcl-2, p53) in HUH-7 cells and caused cell cycle arrest at G0/G1 phase in HCT-116 cells. Molecular docking further supported AMM's apoptosis activity. These results position A. monosperma as a rich source of bioactive polyphenols and antioxidants, with AMM showing promise as a therapeutic agent, especially for colorectal cancer.

Therapeutic resistance to chemotherapy or radiotherapy is a significant issue in several cancers, including head and neck squamous cell carcinoma (HNSCC). One pathway associated with therapeutic resistance is the NFκB pathway, which promotes survival in response to the cytokine TNFα, a key mediator of chemotherapy and radiotherapy-induced cytotoxicity. However, direct targeting of the NFκB pathway is associated with significant toxicity and thus targeting the regulation of this pathway is a promising therapeutic target. We recently demonstrated that the USP14/UCHL5 inhibitor b-AP15 inhibits NFκB activity, inhibiting proliferation and inducing apoptosis in HNSCC cells. Furthermore, b-AP15 treatment sensitised HNSCC cells to the cytotoxic effects of TNFα, as well as TNF-inducing radiation treatment. Here, we investigated if b-AP15 sensitised HNSCC cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a cancer selective member of the TNF family. b-AP15 treatment sensitised HNSCC cells to TRAIL treatment. Mechanistically, we show that b-AP15 induced expression of the TRAIL receptor Death Receptor 5 (DR5)/TRAIL Receptor 2 (TRAILR2), which was required for b-AP15-mediated TRAIL sensitisation. b-AP15 induced reactive oxygen species (ROS) and activated the JNK signalling pathway and both ROS and JNK signalling were required for the induction of DR5 expression and TRAIL sensitisation. We further show that b-AP15-mediated reduction of the NFκB-dependent gene XIAP induced DR5 expression and TRAIL sensitisation and that combination between b-AP15 and IAP antagonists was synergistic in HNSCC cells in vitro. Our data further define the mechanism of b-AP15-mediated cytotoxicity and highlight potential combination treatments that warrant further exploration in pre-clinical studies in HNSCC.

Medicago varia Martyn. exhibited enhanced cold tolerance that was correlated with coordinated adjustments in root xylem structure, modulation of cell wall components, and reprogramming of secondary metabolism, suggesting an integrated adaptive mechanism linking structure, composition, and metabolism. Alfalfa, as one of the most valuable perennial forage crops, is cultivated worldwide. However, its productivity is being threatened by extreme cold events. This study tried to understand the mechanisms of Medicago varia Martyn. (MvM) and Medicago sativa L. (MS) to combat cold climate through chemical, anatomical, spectral, and metabolic analysis. The results showed that MvM had higher content of neutral detergent soluble (74.60%) and soluble proteins (0.15 mg/g), yet lower content of cellulose (10.45%) as compared to MS. Additionally, fourier transform infrared spectroscopy analysis also confirmed differences in functional groups associated with cellulose, hemicellulose, and lignin between MS and MvM. The smaller diameter and higher density of vessel in MvM were consistent with anatomical traits predicted to enhance hydraulic safety under freeze-thaw stress. Metabolomic profiling identified 831 and 604 differentially accumulated metabolites in MvM roots at flowering and senescence stages, with significant enrichment in pathways related to isoflavonoid biosynthesis, arginine/proline metabolism, and tryptophan metabolism. Key metabolites, such as Calceolarioside B, hydroxytyrosol, and Medicarpin, were markedly up-regulated in MvM. Hierarchical clustering highlighted species-specific accumulation of phenylpropanoids and alkaloids in MvM. These findings suggested that the enhanced cold tolerance of MvM might involve in coordinated structural adjustments in root xylem, modulation of cell wall composition, and reprogramming of secondary metabolism. This study provided new insights into the integrative mechanisms of cold adaptation in alfalfa and supported the development of cold-resistant varieties for cultivation in high-latitude regions.

Complex tissue architecture is achieved through multiple rounds of morphological transitions. Here, we analyzed cellular flows and tissue mechanics during avian skin development by employing chicken and transgenic quail skin explant models. We demonstrate how novel cellular flows initiate chemo-mechanical circuits that guide epithelial protrusion, folding, invagination, and spatial cell fate specification. During initial feather bud formation, stiff dermal condensates protrude vertically from the locally softened epithelial sheet. As the bud elongates, it stretches the epithelial cells at the base, thus mechanically activating YAP, which causes the epithelial sheet to fold downward and form a stiff cylindrical wall that invaginates into the skin. This stiff epithelial tongue is essential for the compaction and formation of the tightly packed dermal papillae. These topological transformational events are mechanically interconnected, and the completion of one circuit initiates the next. In contrast, during scale development, the rigid epithelial sheet restricts dermal cell flows, preventing further topological transformation. Based on these findings, we developed a topological transformation model describing how this process enabled the evolution of feather follicles from scales.

This study aimed to determine whether fatty acids (FAs) may affect the function of the early porcine placenta. First, the expression of FA transporters (CD36, SLC27A) in conceptuses and placentae of days 10-11, 12-13, 15-16, 18-20, 25, and 30 pregnant gilts (n = 5-8 per group) was examined using Real-time PCR, Western blot, and immunohistochemistry. Then, primary trophoblast (pTr) cells from days 15-16 conceptuses were exposed to n-6 and n-3 polyunsaturated FAs (PUFAs) to study prostaglandin (PG) synthesis and the expression of genes related to FA action, angiogenesis, steroidogenesis, and lipid transport. Furthermore, pTr cell proliferation and adhesion in response to PUFAs were determined colorimetrically. Increased mRNA expression of CD36, SLC27A1, and SLC27A2 was detected in days 18-25 placentae compared with days 10-13 conceptuses. SLC27A4 and SLC27A6 expression was greater in days 10-11 spherical than in days 15-16 elongated conceptuses. SLC27A1, SLC27A4, and SLC27A6 were localized at the placenta-endometrium interface. PUFAs of n-6 series elevated PGE2 and PGI2 synthesis, whereas n-3 PUFAs stimulated PGE2 but inhibited PGI2 output. All PUFAs up-regulated the mRNA expression of CPT1A, a rate-limiting enzyme of FA β-oxidation. Moreover, docosahexaenoic acid (DHA) increased FABP5, SLC27A4, LDLR (lipoprotein receptor), and proangiogenic ANGPT1 and ANGPTL4 mRNA expression. DHA and arachidonic acid stimulated pTr cell proliferation, while linoleic and eicosapentaenoic acids increased cell adhesion. These results are the first demonstrating dynamic changes of FA transporter expression in peri-implantation conceptuses and developing placentae of the pig and indicate FA uptake by the early placenta. Furthermore, PUFAs may support placenta development by modulating gene expression, increasing PGE2 level, and promoting trophoblast cell viability and adhesion.

Bone remodelling is essential for maintaining skeletal integrity by preserving the balance between bone formation and resorption, with excessive osteoclast activity contributing to osteoporosis. Osteocytes act as central regulators of osteoclastogenesis through mechanically sensitive paracrine signals, yet the influence of osteoblasts and their mesenchymal precursors remains less defined. Extracellular vesicles (EVs) have recently emerged as mediators of bone cell communication, although their role in osteoclast regulation are still underexplored. This study demonstrates that mesenchymal-derived bone cells inhibit osteoclastogenesis through an EV-dependent mechanism shaped by their differentiation stage and mechanical environment. Mechanically stimulated osteocyte-derived EVs showed the strongest anti-catabolic response. Notably, we identify miR-150-5p as a mechano-responsive miRNA enriched within osteocyte EVs, capable of inducing a dose-dependent reduction in osteoclastogenesis. Transcriptomic analyses reveal that EV treatment and miR-150-5p delivery induce substantial transcriptional changes in osteoclast precursors, including downregulation of shared target genes linked to bone remodelling. Overall, we highlight mechanically activated osteocytes as key regulators of osteoclastogenesis through an EV-mediated mechanism, in which miR-150-5p represents a promising candidate contributor within the broader EV cargo landscape, highlighting their potential for future cell-free therapeutic strategies.

Glioblastoma is an aggressive primary brain tumor marked by rapid growth, invasiveness, poor prognosis, and an over 90 % tumor recurrence rate. Current radiation and chemotherapy treatments are limited by non-selectivity and toxicity, creating a need for safer complementary treatments. Historically, natural health products (NHPs) have been used medicinally across cultures for their anti-inflammatory and antioxidant effects. More recently, they have gained recognition for their selective, non-toxic properties in cancer treatment, suggesting their potential as adjuncts to conventional therapies. Black maitake (Grifola frondosa) extract, a well-tolerated NHP with known immunomodulatory properties, has demonstrated anticancer effects in breast cancer models. This study investigates the ability of Black Maitake Odaira Extract - Prothera (BMOE; a trade name of the extract manufactured by Shogun Maitake Canada, London, ON) to induce cell death in the U-87 MG glioblastoma cell line using 2D and 3D models, alone and in combination with the standard chemotherapy: temozolomide (TMZ). Apoptosis was assessed via Hoechst 33,342, annexin V, and propidium iodide staining, along with morphological analyses. Mitochondrial depolarization was measured using TMRM, cell migration was assessed via wound-healing assays, and structural integrity was evaluated using 3D spheroids. BMOE, alone and with TMZ, induced dose-dependent apoptosis, mitochondrial depolarization, and impaired glioblastoma cell migration. BMOE also disrupted 3D spheroid structures and promoted nuclear condensation, consistent with apoptotic processes. Most notably, BMOE significantly enhanced the anti-cancer effects of TMZ. These findings support the potential of BMOE as a complementary therapy that enhances the efficacy of current glioblastoma treatments.

Cytokine-mediated cross-talk between immune cells and fibroblasts is a driver of excessive ECM accumulation during fibrosis. In this study, we used a 3D in vitro model of a connective tissue to discern the roles of three pro-inflammatory cytokines; TNF-α, IL-18 and IL-1β, alone, and in combination with TGF-β1 to simulate the fibrotic environment. Ring-shaped tissues were formed by seeding human fibroblasts into circular molds of agarose, wherein the cells self-assembled, formed a 3D tissue and synthesized de novo a collagen-rich ECM. Cytokine treated tissues were analyzed at days 7 and 14 by histology and measured for thickness, collagen, DNA and strength and stiffness by tensile testing. Despite their pro-inflammatory nature, none of the cytokines increased collagen alone or in combination with TGF-β1. TNF-α and IL-1β reduced collagen, tissue strength and stiffness, and altered tissue morphology. When combined with TGF-β1, TNF-α and IL-1β counteracted TGF-β1-mediated increases in collagen, strength, and stiffness. In contrast, IL-18 had minimal effects alone or when combined with TGF-β1. These data suggest that IL-18 has no effect on fibroblast activation, whereas TNF-α and IL-1β may modulate TGF-β1's effects. This 3D model of a human collagen-rich tissue can help define cytokine-mediated cross-talk between immune cells and fibroblasts.

This study aims to investigate the role of lactylation and m6A modification-related genes in the tumor microenvironment and immunotherapy for hepatocellular carcinoma (HCC) patients. RNA-sequence data and corresponding clinical information of HCC were obtained from the TCGA and ICGC datasets. LASSO Cox regression analysis was implied to construct a lactylation-m6A related prognostic model. The 7-gene signature was established and effectively stratified patients into high- and low-risk groups. Further analysis revealed significant differences between the two risk groups in terms of tumor microenvironment, expression levels of immune checkpoint genes, and drug responsiveness. Specifically, the high-risk group exhibited increased immune cell infiltration, lower IC50 values for several drugs including 5-fluorouracil, afatinib, crizotinib, cediranib, taselisib, and staurosporine; Whereas the low-risk group displayed reduced stromal component proportions and better responses to entinostat, irinotecan, KRAS inhibitors, cisplatin, axitinib, and topotecan. Functionally, knockdown of TCOF1 and HDAC1 significantly attenuated the migration and invasive capacity of Huh-7cells. The lactylation-m6A related prognostic model exhibited robust predictive efficiency in HCC. TCOF1 and HDAC1 may be promising tumor biomarkers for HCC and more researches are needed to validate these results.

Preeclampsia (PE) is a pregnancy-specific hypertensive disorder that could lead to serious maternal and fetal complications, yet early identification of women at risk remains challenging because reliable biomarkers are limited. Here we show that generating relatively stable cell-free DNA (cfDNA) fragmentomic metrics, including transcription start site (TSS) coverage, TSS score, and Gini coefficient, required 600 million whole-genome sequencing reads of plasma cfDNA. These metrics exhibited observable differences among genes with varying expression levels in blood cells and placental tissues. In a cohort of 1,058 pregnant women, cfDNA fragmentomics could distinguish pregnancies that subsequently developed PE. When integrated with maternal risk factors, predictive models in two independent test sets achieved mean area under the curves of 0.903 and 0.850 for early-onset and late-onset PE, respectively, with sensitivities of 0.731 and 0.607 at a 10% false positive rate. Importantly, these models also performed well in samples collected before or at 16 weeks of gestation, supporting the potential of cfDNA fragmentomics in early PE risk assessment.

A minority of 8C-like cells (8CLCs) expressing zygotic genome activation genes are captured in naïve embryonic stem cells (ESCs). However, how human ESCs transition into 8CLCs remains unknown. Here, we show that NELFA is dispensable in human primed ESCs but critical for naïve pluripotency, and its overexpression promotes the primed-to-naïve conversion. Notably, NELFA robustly induces 8CLCs that express human ZGA-specific genes in naïve ESCs. NELFA-induced human 8CLCs can contribute to both embryonic and extraembryonic developmental potential in the interspecies human-mouse chimera assay. Importantly, we also discovered that TP53 can activate the human 8CLCs state, and NELFA might be an upstream regulator of TP53. Furthermore, TP53 and NELFA are both essential for the complete transition to the 8CLC state. We also demonstrate that sustained NELFA overexpression in primed ESCs directs neural lineage specification. Thus, NELFA establishes and maintains naïve pluripotency and drives the 8CLC state, providing insights into early human embryogenesis.

Peripheral nerve injury (PNI), a common clinical condition, heavily relies on the functional condition of Schwann cells (SCs) for effective repair. Emerging evidence shows that excessive pyroptosis of SCs hinders neural regeneration. Low-intensity pulsed ultrasound (LIPUS), a non-invasive physical therapy, shows potential in tissue repair; however, its role in regulating SCs pyroptosis remains unclear. This study explores the mechanism by which LIPUS improves PNI recovery by reducing SCs pyroptosis. We find that LIPUS significantly enhances motor function recovery, promotes axonal regeneration and remyelination, and decreases gastrocnemius muscle atrophy through a rat sciatic nerve crush model. LIPUS biomechanically mitigates mitochondrial dysfunction in SCs, thereby suppressing the NLRP3/Caspase-1/GSDMD-N pyroptosis signaling pathway. This inhibition reduces IL-1β and IL-18 release, boosting SCs proliferation, migration, and clearance of myelin debris, collectively fostering a regenerative microenvironment that supports axonal regrowth, remyelination, and functional recovery. Our results demonstrate a mechanobiological mechanism by which LIPUS promotes peripheral nerve regeneration through alleviating pyroptosis, providing a promising therapeutic approach for PNI.

Accumulating evidence demonstrates that the silencing of tumor suppressor genes by aberrant DNA methylation contributes to the initiation and progression of ovarian cancer (OC), while the systemic methylation profile and the key driver methylation events need to be further explored. Here, by analysing public databases and our resources, we identified the hypermethylation of ZNF154 promoter as a key driver of OC malignancy, which was mediated by the DNA methyltransferase complex DNMT1/UHRF1. Using CRISPR/dCas9-TET1CD, a tool for targeted demethylation, we successfully decreased the methylation level of ZNF154 promoter and reactivated ZNF154 expression, which in turn inhibited the proliferation, migration, and invasion of OC cells. Mechanistically, ZNF154 interacted with KAP1 and directly bound to the ROMO1 promoter, transcriptionally repressing ROMO1 expression, thereby reducing MMP2 and phosphorylated ERK to impede OC progression. Clinically, ZNF154 hypermethylation was correlated with its reduced expression and poor prognosis in OC patients. These findings underscore a pivotal role of aberrant ZNF154 methylation in OC pathogenesis and highlight its potential as both a therapeutic target and a prognostic biomarker for OC patients.

The practical application of Fe-N-C catalysts in proton exchange membrane fuel cells is fundamentally constrained by the inherent activity-stability trade-off. Here, we propose a "repair-and-upgrade" engineering strategy that not only repairs pyrolysis-induced defects through carbon and nitrogen supplementation but also evolves conventional FeN4 moieties into stabilized FeN5 configurations via an in situ constructed carbon bilayer. The axial nitrogen modulates the electronic structure of Fe center to enhance catalytic activity, while the adaptive interlayer spacing of the N-linked carbon bilayer compensates for fluctuations in the axial Fe─N bond length during catalysis, therefore anchoring the Fe active sites. When integrated into membrane electrode assemblies, the catalyst delivers a high peak power density of 1221 mW cm-2 and exhibits exceptional durability, retaining over 85% of its initial power density after 10,000 cycles in H2-O2 and showing negligible decay over 45 h at 0.6 V in H2-air tests. This work presents a novel design strategy for stable single-atom catalysts, centered on creating an adaptive local environment that ensures exceptional electrocatalytic stability.

Executive function is an essential cognitive domain for typical human behavior which is disrupted in neurodevelopmental and neurodegenerative disorders, but little is known about its underlying molecular basis. To address this, we perform genome-wide association studies (GWAS) using three different measures of executive function in UK Biobank (N = 84,238) and NIHR BioResource's Genes and Cognition (N = 9932) study participants, followed by a meta-analysis. The trail-making alphanumeric (TMA) measure is the most heritable phenotype (h²=7-26%), associated with 18 independent loci that exhibit a similar direction of effect in both cohorts. Across these loci, in-silico follow-up implicates 178 genes, of which NT5DC2 and RP11-579E24.2 are independently replicated prior to meta-analysis. TMA is linked to pan-cerebral differences in brain structure, with brain-enriched genes showing a biphasic expression profile from early development through to later life. Our data implicate specific cell types, histone modifications and butyrophilin immunoglobulin family proteins as potential targets for promoting cognitive resilience.