Hair follicle stem cells (HFSCs) can quickly activate and migrate to the wound site, differentiating into epidermal stem cells to facilitate early epithelialization. During the wound healing process, microRNAs (miRNAs) function coordinately. Chinese medicine borneol is derived from the Cinnamomum camphora plant, and its principal component, L-borneol, is renowned for its potential in facilitating skin wound healing. However, it remains unclear whether L-borneol can stimulate HFSCs to differentiate into epidermal cells or whether miRNAs are involved in this process. In the current study, HFSCs were isolated from the vibrissae of rats and identified based on the expression of CD34, Integrin-β1 and keratin type 1 cytoskeletal 15(CK15). We observed that stimulation with L-borneol significantly increased the differentiation marker K14 in HFSCs, suggesting that L-borneol could promote the differentiation of HFSCs into basal layer cells. On this basis, we transfected and confirmed that rno-miR-127 inhibitor could promote the differentiation of HFSCs. Furthermore, we demonstrated that PODXL2 is a target gene of rno-miR-127 through dual-luciferase reporter assays and confirmed that the rno-miR-127 mimic could inhibit the expression of PODXL2. To further elucidate the targeting relationship, we constructed the siPODXL2 fragment using siRNA technology, demonstrating that reducing PODXL2 expression can inhibit the differentiation of HFSCs into basal layer cells. Finally, a rat full-thickness skin defect model illustrated L-borneol-mediated negative regulation of PODXL2 by rno-miR-127, promoting skin injury repair through HFSCs.

Acute myeloid leukemia (AML) is an aggressive and heterogeneous hematological malignancy with a low survival probability and limited therapeutic options. Although galectin-1 (LGALS1) has been implicated in tumor cell survival and immune evasion in solid tumor, its role in AML is still unclear. In this study, we found that LGALS1 presents prominent upregulation in AML patients at both mRNA and protein levels compared with the control samples. Bioinformatics analysis indicated that high expression of LGALS1 is a significant unfavorable prognostic factor for overall survival in AML, correlating with adverse clinical and genetic features as well as immune cell infiltration. Depletion of LGALS1 in AML cells impeded cell proliferation, induced apoptosis and promoted myeloid differentiation. Treatment with OTX008, an LGALS1 inhibitor, markedly diminished the viability of primary malignant bone marrow cells from AML patients. Notably, LGALS1 expression was significantly reduced exclusively in AML-M5 patients after treatment, which may be due to its higher expression in AML-M5 subtype compared to other FAB subtypes. In summary, our findings indicate that LGALS1 could serve as an independent prognostic risk factor and a promising therapeutic target in AML, providing novel insights into AML pathogenesis and laying the foundation for the development of new therapeutic strategies.

The pathological process of osteoporosis involves accelerated bone resorption and a decline in bone formation, among which the disruption of the balance between adipogenic and osteogenic differentiation in bone marrow mesenchymal stem cells (BMSCs) is a crucial part. Cellular repressor of E1A-stimulated genes 1 (CREG1), a small glycoprotein, is mainly localized to the endosomal-lysosomal compartment and is associated with the regulation of mitophagy and cell differentiation. However, its roles in BMSCs osteogenic differentiation and skeletal degenerative disorders, including osteoporosis, are poorly understood. We previously identified CREG1 as being highly expressed in the bone marrow through database analysis and found that its expression increased in the process of BMSCs osteogenic differentiation. In the present study, we demonstrated that the expression of CREG1 was reduced in osteoporosis patients and animal models, and the overexpression of CREG1 contributed to higher bone mass compared with ovariectomy (OVX)-induced bone loss models. Further research revealed that the knockdown of CREG1 inhibited the osteogenic differentiation of BMSCs, while CREG1 overexpression promoted this process. Additionally, we found that CREG1 overexpression was accompanied by an increase in mitophagy levels, and the osteogenic differentiation induced by this overexpression was blocked when mitophagy was inhibited, indicating that CREG1 promoted osteogenic differentiation through inducing mitophagy. Therefore, our findings demonstrated that CREG1 is involved in regulating the osteogenic differentiation of BMSCs, thereby providing new therapeutic targets and pathways for the treatment of osteoporosis.

Development of type 1 conventional dendritic cells (cDC1s) underlies the capacity to generate antiviral and antitumor immune responses. Here, we identify the basis for cDC1 development from its earliest progenitors, determining the hierarchy of several required transcription factors and uncovering a series of mandatory cis interactions between constituent enhancers within the Irf8 superenhancer. We produced in vivo mutations of two C/EBPα binding sites that comprise the Irf8 +56-kilobase (kb) enhancer that markedly reduced IRF8 expression in all myeloid progenitors and impaired cDC1 development. These sites did not bind RUNX1 or RUNX3, and C/EBPα expression was instead regulated by their action at the Cebpa +37-kb enhancer, placing RUNX factors upstream of Cebpa in regulating Irf8. Last, we demonstrate that cis interactions between the +56-kb Irf8 enhancer and the previously reported +41- and +32-kb Irf8 enhancers are mandatory in the sequential progression of these stage-specific constituent elements.

Background: Vascular smooth muscle cells (vSMCs), the predominant cell type in the aortic wall, play a crucial role in maintaining aortic integrity, blood pressure, and cardiovascular function. vSMC contractility and function depend on smooth muscle alpha-actin 2 (ACTA2). The pathogenic variant ACTA2 c.536G>A (p. R179H) causes multisystemic smooth muscle dysfunction syndrome (MSMDS), a severe disorder marked by widespread smooth muscle abnormalities, resulting in life-threatening aortic disease and high-risk early mortality from aneurysms or stroke. No effective treatments exist for MSMDS. Methods: To develop a comprehensive therapy for MSMDS, we utilized CRISPR-Cas9 adenine base editing to correct the ACTA2 R179H mutation. We generated isogenic human induced pluripotent stem cell (iPSC) lines and humanized mice carrying this pathogenic missense mutation. iPSC-SMCs were evaluated for key functional characteristics, including proliferation, migration, and contractility. The adenine base editor (ABE) ABE8e-SpCas9-VRQR under control of either a SMC-specific promoter or a CMV promoter, and an optimized single guide RNA (sgRNA) under control of U6 promoter were delivered intravenously to humanized R179H mice using adeno-associated virus serotype 9 (AAV9) and phenotypic outcomes were evaluated. Results: The R179H mutation causes a dramatic phenotypic switch in human iPSC-SMCs from a contractile to a synthetic state, a transition associated with aneurysm formation. Base editing prevented this pathogenic phenotypic switch and restored normal SMC function. In humanized mice, the ACTA2R179H/+ mutation caused widespread smooth muscle dysfunction, manifesting as decreased blood pressure, aortic dilation and dissection, bladder enlargement, gut dilation, and hydronephrosis. In vivo base editing rescued these pathological abnormalities, normalizing smooth muscle function. Conclusions: This study demonstrates the effectiveness of adenine base editing to treat MSMDS and restore aortic smooth muscle function. By correcting the ACTA2 R179H mutation, the pathogenic phenotypic shift in SMCs was prevented, key aortic smooth muscle functions were restored, and life-threatening aortic dilation and dissection were mitigated in humanized mice. These findings underscore the promise of gene-editing therapies in addressing the underlying genetic causes of smooth muscle disorders and offer a potential transformative treatment for patients facing severe vascular complications.

The switch from precursor cell proliferation to onset of differentiation in adult stem cell lineages must be carefully regulated to produce sufficient progeny to maintain and repair tissues, yet prevent overproliferation that may enable oncogenesis. In the Drosophila male germ cell lineage, spermatogonia produced by germ line stem cells undergo a limited number of transit amplifying mitotic divisions before switching to the spermatocyte program that sets up meiosis and eventual spermatid differentiation. The number of transit amplifying divisions is set by accumulation of the bag-of-marbles (Bam) protein to a critical threshold. In bam mutants, spermatogonia proliferate through several extra rounds of mitosis and then die without becoming spermatocytes. Here, we show that a key role of Bam for the mitosis to differentiation switch is repressing expression of Held Out Wings (how), homolog of mammalian Quaking. Knockdown of how in germ cells was sufficient to allow spermatogonia mutant for bam or its partner benign gonial cell neoplasm to differentiate, while forced expression of nuclear-targeted How protein in spermatogonia wild-type for bam resulted in continued proliferation at the expense of differentiation. Our findings suggest that Bam targets how RNA for degradation by acting as an adapter to recruit the CCR4-NOT deadenylation complex via binding its subunit, Caf40. As How is itself an RNA-binding protein with roles in RNA processing, our findings reveal that the switch from proliferation to meiosis and differentiation in the Drosophila male germ line adult stem cell lineage is regulated by a cascade of RNA-binding proteins.

The efficient differentiation of adult gastric stem cells into specific epithelial cell types is crucial for gastric homeostasis. Although it is well appreciated that the niche plays a critical role in gastric epithelium cell differentiation, the relevant molecular factors and the underlying regulatory mechanisms remain poorly understood. In this study, by combining the knowledge of the niche cells obtained from single-cell RNA sequencing and manipulation of signaling pathways, we achieved effective differentiation of various gastric epithelial cell types in mouse and human gastric organoids. These in vitro differentiated cells showed a similar gene expression profile to those in gastric tissues. Specifically, BMP4 signaling stimulates pit cell and parietal cell differentiation. Furthermore, BMP4 and EGF signaling cooperate to enhance pit cell differentiation, whereas inhibition of TGF-β and BMP4 signaling promotes chief cell differentiation. We demonstrated that Zbtb7b is a novel regulator controlling pit cell differentiation. In addition, BMP4, together with the small molecule Isoxazole 9, promotes parietal and enteroendocrine cell differentiation. Our data also revealed the different requirements of parietal and chief cell differentiation between mouse and human. Together, our findings provide a mechanistic insight into gastric epithelial cell differentiation and uncover its similarities and differences between mouse and human, laying a foundation for future investigation and potential clinical use of gastric organoids.

Metformin's topical application has proven to be a therapeutic wound healing dressing via pro-angiogenic, anti-inflammatory, autophagy-promoting, and antibacterial effects. The systematic review article aimed to evaluate the available evidence (from both preclinical and clinical studies), in order to better understand its therapeutic potential in wound care. We performed a systematic literature search in PubMed, Scopus, Web of Science, Embase, and Cochrane databases till January 2025. A total of 26 studies included in final analysis according to inclusion criteria. The majority of preclinical investigations demonstrated accelerated wound healing with characterized of increased wound closure and collagen deposition, elevated pro-angiogenic markers (e.g., VEGF, CD31, and α-SMA), suppression of pro-inflammatory cytokines, and induction of autophagy signaling. Notably, biomaterial-based delivery platform applications such as hydrogels and nanofibers greatly magnified these therapeutic effects. While encouraging results in in-vitro, in-vivo and animal models have been documented, clinical studies remain limited in scope. In fact, this large disparity between preclinical results and limited clinical evidences obviously highlights the urgent need for properly designed human trials to determine safety, efficacy, and best delivery modalities of topical metformin. Collectively, topical metformin is a novel and potentially valuable addition to wound treatment cares, which could be subjected to further clinical study.

The aim of this study was to explore the prognostic value of mean corpuscular volume (MCV) in newly diagnosed aplastic anemia (AA) patients treated with cyclosporine A (CsA) plus androgen or CsA alone. The clinical data of 181 newly diagnosed patients with aplastic anemia from April 2008 to September 2020 in the Affiliated Hospital of Xuzhou Medical University were retrospectively analyzed. According to the MCV levels, the patients were divided into a high-MCV group (107/181) and a normal-MCV group (74/181). We investigated the effect of MCV outcomes in patients with AA. Between the high-MCV and normal-MCV groups, neutrophil count, red blood cell count, platelet count, reticulocyte count, disease severity, lymphocytes, and granulocytes were significantly different (P < 0.05). The overall response rates (CR + PR) were 69.16% and 59.46% in the high-MCV and normal-MCV groups, respectively. The duration of response was not significantly different between MCV groups. The high-MCV patients had an improved 5-year overall survival and progression-free survival compared to the normal-MCV patients (94.40% vs. 68.10%; 71.80% vs. 60.30%, P < 0.001). Regarding hemogram restoration, the leukocyte, neutrophil, hemoglobin, and platelet recovery was accelerated in the high-MCV group (P < 0.05). Furthermore, MCV levels were positively correlated with reticulocyte count, reticulocyte percentage, high-fluorescence reticulocyte, medium-fluorescence reticulocyte, and immature reticulocyte fraction; on the other hand, MCV levels were negatively correlated with low fluorescent reticulocyte. In conclusion, aplastic anemia patients with a high MCV were a better prognostic factor, and the patients with high MCV may have better residual bone marrow hematopoietic function than those with normal MCV.

With increasing data on graft-versus-tumor (GVT) effect in neuroblastoma, we aimed to evaluate the possibility of haploidentical stem cell transplantation (HSCT) as a technique to induce GVT and thereby improve outcomes in high-risk neuroblastoma.

Mucosal-associated invariant T (MAIT) cells are diminished in various liver diseases, but the underlying mechanism remains unclear. This study aimed to investigate the characteristics and underlying mechanisms of MAIT cell depletion in HBV-related cirrhosis.

Ordered intermetallic PtCo alloys are promising candidates for next-generation low-Pt catalysts in proton exchange membrane fuel cells (PEMFCs) due to their high activity and stability originating from ligand and strain effects. However, the influence of the ordered phase on the surface structure, especially after Pt-rich shell formation, remains poorly understood. In this study, we systematically investigated the structural and electrochemical properties of PtCo catalysts with varying degrees of ordering, prepared by controlling the annealing temperature and time. We combined X-ray diffraction (XRD), X-ray absorption spectroscopy (XAS), scanning transmission electron microscopy (STEM), and pair distribution function (PDF) analysis to elucidate the correlation between the ordering degree, the atomic structure, and the electrochemical performance. For the first time, our detailed X-ray total scattering measurements revealed the true structural characteristics of PtCo alloys, indicating that the phase types and their relative contents vary significantly with the ordering degree. The ordered PtCo catalysts develop a Pt-rich surface layer with anisotropic strain, featuring contracted Pt-Pt distances along the surface and elongated distances across the surface, which likely contributes to its enhanced ORR activity and stability compared to their disordered counterparts. The electrochemical studies and PDF analysis suggested that the ordering transition occurs concurrently with particle growth, leading to an abrupt increase in the ORR activity at 350 °C before forming a long-range ordered phase, suggesting that local structural changes at the particle surface play a crucial role in enhancing the ORR activity. Further, operando XAS studies confirm lesser Pt-oxidation and Pt-OH formation for ordered structures than disordered ones. We believe that our findings provide new insights into the relationship between the ordering degree, particle growth, and catalytic properties of PtCo catalysts, offering guidance for the design of high-performance electrocatalysts with optimized surface structures.

Induced pluripotent stem cells induced hepatocytes (iHeps) are widely used in modeling human liver diseases and as a potential cell source for replacement therapy. However, most iHeps are relatively immature and challenging to maintain for long-term in vitro culture.

Targeting cancer stem cells (CSCs)-mediated aggressive features of non-small cell lung cancer (NSCLC) is a promising anticancer approach. This can be accomplished via suppressing critical mediators, such as functional mitochondria, aldehyde dehydrogenase (ALDH)1A, and telomere protectors (telomerase reverse transcriptase (TERT) and telomere repeat binding factor (TRF)1).

Acute lymphoblastic leukemia (ALL) is a disease of lymphoid progenitor cells arising in the bone marrow and extramedullary sites. While it is the most common pediatric cancer, ALL is a rare disease overall, with approximately 6500 new cases diagnosed in the United States, in 2024. Current treatment relies on multiagent chemotherapy administered over 2-3 years, resulting in long-term survival in 80%-90% in pediatric patients compared to 40%-50% in adult patients, depending upon patient- and disease-specific characteristics.

Enterovirus A71 (EV-A71) is a causative agent of hand, foot, and mouth diseases. EV-A71 infections may result in severe neurological complications in children. Although several receptors or attachment molecules for EV-A71 have been identified, EV-A71 can still infect host cells even after blocking these receptors with antibodies. We have previously identified plasminogen (PLG), a circulating zymogen of plasmin, as a cell membrane-associated EV-A71-interacting glycoprotein. We confirmed that anti-PLG antibodies could reduce the binding of EV-A71 to RD cells as anti-SCARB2 and anti-nucleolin. Knockdown of PLG reduced EV-A71 binding to RD cells, and preincubation of PLG with EV-A71 increased virus binding. Enzyme-linked immunosorbent assay and surface plasmon resonance assays demonstrated the direct binding of PLG to EV-A71. We further evaluated the biological characteristics of EV-A71-infected PLG knockout (heterozygous) and wild-type mice. We found that the clinical scores and mortality of WT mice were higher than those of PLG-knockout mice after EV-A71 infection. The viral loads in the spinal cord of PLG knockout mice were lower than those in WT mice 6 days post-infection. EV-A71-associated cytokines such as IL-1β, IL-6, MCP-1, IL-10, and IFN-γ were investigated. Serum IL-10 and MCP-1 expression were significantly higher in EV-71-infected WT mice than in PLG knockout mice, and MCP-1 may be one of the critical chemokines that induce intense inflammation and chemoattracts leukocytes. Our findings reveal a possible role for PLG in EV-A71 infection/pathogenesis and shed light on developing novel therapeutic approaches and drugs to prevent EV-A71 infection.IMPORTANCEUnderstanding the pathogenesis of enterovirus A71 (EV-A71) for developing novel drugs or therapeutic approaches has always been a significant issue. In this study, we demonstrated the interactions between plasminogen (PLG) and EV-A71, characterized the biological effects of EV-A71-infected PLG knockout mice, and evaluated their immune response. We found that EV-A71 caused more severe tissue damage than PLG knockout mice in skeletal muscle, spinal cord, and brain stem. Higher virus protein was observed in these tissues of WT mice. The reduced clinical scores, mortality, and cytokine expression suggested PLG may be involved in EV-A71 infection-induced cytokine storm. The findings and animal model in the current study provide the new drug target for anti-EV-A71 drug discovery.

Synthetic developmental biology uses engineering approaches to understand multicellularity with goals ranging from recapitulating development to building synthetic organisms. Current approaches include engineering multicellular patterning, controlling differentiation, and implementing cooperative cellular behaviors in model systems. Synthetic biology enables these pursuits by providing tools to control cell behavior. Mouse embryonic stem cells (mESCs) offer a well-studied and genetically tractable pluripotent model for pursuing synthetic development questions. However, there is minimal characterization of existing synthetic biology tools in mESCs. Here, we characterize three small molecule and two cell contact-inducible systems for gene expression in and differentiation of mESCs. We show that small molecule and cell-contact inducible systems work reliably and efficiently for controlling expression of arbitrary genetic payloads. We identify how these systems function differently across model differentiations. Furthermore, we show that these systems can drive direct differentiation of mESCs into neurons. Each of these systems can be used on their own or in combination, opening many possibilities for studying developmental principles with high precision.

During cortical development, neural stem/precursor cells (NS/PCs) sequentially produce neurons, astrocytes, and oligodendrocytes. Before producing these cells, human (h) NS/PCs undergo prolonged self-renewal to form a larger cortex than other mammals, although the mechanisms are mostly unknown. Here, we performed a gene knockout screen using the CRISPR/Cas9 system to search for genes involved in hNS/PC self-renewal. We identified RMND5A, encoding an E3 ubiquitin ligase, among the candidate genes. We further demonstrated that knockdown of RMND5A decreased proliferation and promoted neuronal differentiation of hNS/PCs through the activation and suppression of the Wnt and mTOR signaling pathways, respectively. Taken together, our findings suggest that RMND5A participates in the maintenance of hNS/PC self-renewal by modulating the Wnt and mTOR signaling pathways. Impact statement During cortical development, human neural stem/precursor cells (hNS/PCs) undergo prolonged self-renewal to form a larger cortex than other mammals, although the mechanisms are mostly unknown. We identified RMND5A, an E3 ubiquitin ligase, as essential for maintaining self-renewal of hNS/PCs, providing valuable insights into the evolutionary expansion of the human brain.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that, in Fig. 3A and B on p. 1145 showing the results of cell invasion and migration experiments, four pairs of overlapping data panels were identified, affecting half the panels shown in the figure, such that these data panels, which were intended to show the results of differently performed experiments, had all apparently been derived from the same original source. Owing to the large number of duplications of data that have been identified in this paper, the Editor of International Journal of Oncology has decided that it should be retracted from the Journal on account of a lack of confidence in the presented data. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [International Journal of Oncology 46: 1141‑1148, 2015; DOI: 10.3892/ijo.2014.2809].

Harnessing natural developmental programs to repair and replace damaged organs represents promising approaches in regenerative medicine. However, effective strategies are still lacking for tissue regeneration in complicated conditions, such as the periodontal bone defect. Here, human dental follicle stem cells (hDFSCs) and their aggregates (hDFSCA) are cultured and characterized, which are formed based on the inherent property of these stem cells self-assembly into compact spheroid-like structures, mimicking mesenchymal condensation in development. A periodontal tissue-specific microenvironment simulation material is then established, human decellularized alveolar bone matrix particles (hDABMPs), which possess favorable physicochemical and biological properties for regenerative use. hDFSCs co-cultured with hDABMPs exhibit improved cell function, and hDFSCA-hDABMP co-aggregates are subsequently constructed, which activate the developmental gene expression in hDFSCA and initiate hypoxic adaptation mechanisms for tissue regeneration. Indeed, hDFSCA-hDABMP co-aggregates significantly promote regeneration after implantation in alveolar bone defects with good biosafety. Interestingly, during the early stages of implantation, hDABMPs enhance hDFSC survival and expansion, thereby providing a sufficient source of cells for tissue regeneration. Collectively, this study reveals a development-inspired, engineered cell-niche co-aggregation strategy for enhancing CA therapeutic potential by simulating tissue-specific microenvironments, offering novel insights for functional tissue regeneration.