Small extracellular vesicles (sEV) derived from mesenchymal stem cells hold promise for anti-skin aging, yet their clinical application is hindered by poor transdermal permeability. Herein, we report an innovative light-controlled sEV-based spherical nucleic acid nanomotor (NM-ESNA). This nanosystem was composed of an sEV core and an MMP1-targeting siRNA shell, forming a 3D penetrative nanostructure. In addition, asymmetrically modified light-responsive gas-generating molecules were integrated into the nanomotor, enabling efficient dermal delivery. The light-controlled and enhanced transdermal delivery guaranteed synergistic anti-skin aging therapy through sEV-mediated paracrine effects and gene therapy targeting MMP1 in the dermis. On the basis of this deep transdermal delivery technology and the synergistic therapy strategy, NM-ESNA demonstrated outstanding anti-skin aging effects in a mouse model. This biocompatible nanosystem (NM-ESNA) enabled light-controlled and deep transdermal delivery, establishing a therapeutic platform with significant potential for sEV-based noninvasive anti-skin aging therapy.

EphA2 receptor tyrosine kinase is overexpressed in many solid tumors and serves as a key driver of tumorigenesis and metastasis. It is highly expressed in glioblastoma multiforme, the most aggressive brain tumor in adults, and in its stem cells [glioblastoma stem cells (GSCs)], which contribute to treatment resistance and tumor relapse. In a previous study, we used the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) procedure, a method for selecting high-affinity nucleic acids to specific targets via iterative selection and amplification, to identify the 2'-fluorinated EphA2-targeting RNA aptamer A40L and a truncated 30-mer derivative, A40s. Both aptamers were able to inhibit GSC growth, stemness, and migration upon EphA2 binding. Here, by integrating computational and experimental methods, the A40s structure was unraveled and its interaction with EphA2 was investigated. Our model offers a blueprint to accelerate the development of optimized A40s variants, advancing next-generation EphA2-targeted anticancer therapies.

Flowering and secondary growth are two important biological processes in plant development, and there is a close association between flowering and xylem expansion in stem and root secondary growth. Here, we report the discovery of NAC secondary wall thickening promoting factor 3 (BnaA9.NST3), a key regulator of secondary cell wall formation in xylem fiber cells that affects B. napus (Brassica napus L.) flowering in a photoperiod pathway-dependent manner. We show that the subfunctionalized BnaA9.NST3 directly binds to the BnaC8.NF-YC9 and Bna.CO promoters to regulate their transcription. BnaC8.NF-YC9 interacts with Bna.CO, forming NF‒CO complexes that regulate Bna.FT transcription, thereby affecting flowering. Moreover, we found that florigen upregulated secondary cell wall biogenesis (SCWB)-related genes including BnaA9.NST3, accelerating SCWB in stems and promoting vascular maturation and adaptation of the shoot system to meet the plant's mechanical support needs during the reproductive phase. Our results provide insights into the connection between flowering and secondary growth and suggest gene targets for cultivating B. napus lines that flower at the optimal time.

Lysine-specific histone demethylase 1A (LSD1) is an epigenetic regulator involved in various biological processes, including metabolic pathways. We demonstrated the therapeutic potential of its pharmacological inhibition in glioblastoma using DDP_38003 (LSD1i), which selectively targets tumor-initiating cells (TICs) by hampering their adaptability to stress. Through biological, metabolic, and omic approaches, we now show that LSD1i acts as an endoplasmic reticulum (ER) stressor, activating the integrated stress response and altering mitochondrial structure and function. These effects impair TICs' oxidative metabolism and generate reactive oxygen species, further amplifying cellular stress. LSD1i also impairs TICs' glycolytic activity, causing their metabolic decline. TICs with enhanced glycolysis benefit from LSD1-directed therapy. Conversely, metabolically silent TICs mantain ER and mitochondrial homeostasis, adapting to stress conditions, including LSD1i treatment. A dropout short hairpin RNA screening identifies postglycosylphosphatidylinositol attachment to proteins inositol deacylase 1 (PGAP1) as a mediator of resistance to LSD1i. Disruptions in ER and mitochondrial balance holds promise for improving LSD1-targeted therapy efficacy and overcoming treatment resistance.

The self-renewal of intestinal stem cells (ISCs) is essential for maintaining intestinal homeostasis and ensuring regeneration of the intestinal epithelium. However, whether small nucleolar RNAs participate in the regulation of ISC self-renewal remains unclear. Here, we identified a small nucleolar RNA (Snora54) that was highly expressed in the nucleolus of ISCs. Snora54 knockout enhanced the self-renewal capacity of ISCs and intestinal regeneration. Mechanistically, in a steady state, highly expressed Snora54 anchored the nucleolar protein Lyar in the nucleolus of ISCs, preventing Lyar from translocation into the nucleoplasm. Thereby, Lyar failed to recruit on the Notch2 promoter region in the nucleoplasm to promote Notch2 transcription, leading to suppression of ISC self-renewal. By contrast, with deletion of Snora54, Lyar translocated to the nucleoplasm of ISCs where it enriched on the Notch2 promoter to initiate its transcription resulting in the activation of Notch2 signaling pathway. Therefore, Snora54 negatively regulates self-renewal of ISCs and gut regeneration via suppression of Notch2 signaling.

The adult mammalian heart has limited regenerative capacity due to the low proliferative ability of cardiomyocytes, whereas embryonic cardiomyocytes exhibit robust proliferative potential. Using single-cell RNA sequencing of embryonic hearts, we identified prothymosin α (PTMA) as a key factor driving cardiomyocyte proliferation. Overexpression of PTMA in primary mouse and rat cardiomyocytes significantly promoted cardiomyocyte proliferation and similarly enhanced proliferation in human iPSC-derived cardiomyocytes. Conditional knockout of Ptma in cardiomyocytes impaired neonatal heart regeneration. AAV9-mediated overexpression of Ptma extended the neonatal proliferative window and showed therapeutic promise for enhancing adult heart regeneration. Mechanistically, PTMA interacted with MBD3, inhibiting its deacetylation activity within the MBD3/HDAC1 NuRD complex. This inhibition increased STAT3 acetylation, which positively regulated STAT3 phosphorylation and activation of its target genes. These findings establish PTMA as a critical regulator of heart regeneration and suggest its potential as a therapeutic target for ischemic myocardial injury.

Zika virus (ZIKV) infection can lead to a variety of clinical outcomes, including severe congenital abnormalities. The phosphatidylserine receptors AXL and TIM-1 are recognized as critical entry factors for ZIKV in vitro. However, it remains unclear whether and how ZIKV regulates these receptors during infection. In this study, we investigated AXL and TIM-1 expression in human lung adenocarcinoma epithelial A549 cells, glioblastoma U87 cells, and embryonic stem cell-derived trophoblasts following ZIKV infection. We found that both the Asian strain FSS13025 and the African strain MR766 of ZIKV downregulate AXL, with a milder effect on TIM-1. We identified several ZIKV proteins, notably envelope (E), NS2A, NS3, and NS4B, that contribute to this downregulation. Notably, treatment with lysosomal inhibitor NH4Cl or the autophagy inhibitor 3-methyladenine mitigated the AXL/TIM-1 downregulation, indicating autophagy's involvement in the process. Importantly, this downregulation facilitates sustained viral replication and promotes viral spread by preventing superinfection and limiting cell death, which is also associated with impaired innate immune signaling. Our findings uncover a mechanism by which ZIKV downregulates entry factors to enhance prolonged viral replication and spread.

Siraitia grosvenorii, commonly known as Monk fruit, is the fruit of S. grosvenorii (Swingle) C. Jeffrey ex A. M. Lu & Zhi Y. Zhang (Jeffrey C. 1980). As a traditional medicinal and edible homologous herb, S. grosvenorii is widely distributed in southern China (Li et al. 2025). In July 2024, wilting plants were found in farmed fields in Longsheng Nationalities Autonomous County (25.714443°N, 110.032812°E), Guilin, Guangxi, China, with an estimated incidence rate of 10 to 30%. Initially the leaves exhibited signs of water loss. The stems and leaves gradually turned yellow and withered, ultimately leading to the death of the entire plant. To isolate the causal agent, 12 small pieces of discolored stem tissues were cut, and then they were surface sterilized by using 75% EtOH for 30s and placed in 2.5% NaOCl for 2 min, and then rinsed three times in sterilized distilled water. These samples were then placed on potato dextrose agar (PDA) and they were incubated at 25℃ in the darkness for 7 days. The hyphal tips were transferred to fresh PDA in order to obtain pure cultures. 26 isolates were obtained, 19 of which had similar morphology. These were isolated from all the symptomatic stems and they were considered to be pathogens, and named L1. After 14 days, the colonies were white, fluffy, with white aerial mycelium, while the reverse side turned dark black, due to the formation of microsclerotia. The hyphae were hyaline and formed whorled branches. The conidia were hyaline, aseptate, cylindrical to oval and measured 4.3 to 6.6 × 1.6 to 3.1 μm in size ( n = 60). The microsclerotia were immersed in the agar, and they were composed of rounded cells and were elongated or irregular in shape. These morphological features were consistent with Verticillium dahliae (Isaac 1967). The L1 isolate was subjected to sequencing of the internal transcribed spacer (ITS), actin (ACT) and glyceraldehyde-3-phosphate dehydrogenase (GPD) sections (Inderbitzin et al. 2011) for molecular identification. The ITS, ACT and GPD sequences (Genebank accession nos. PQ836512, PQ847485 and PQ847486) displayed 99 to 100% (529/531, 475/475, 640/640 bp) similarity to V. dahliae (Genebank accession nos. KX058040, OK513223 and OK513236, respectively). Phylogenetic analysis was based on a combination of ITS, ACT and GPD sequence data using MEGA 11 software to construct a phylogenetic tree with neighbor-joining. In the phylogenetic tree, the combined sequences attributed L1 to V. dahliae. A pathogenicity test was performed by soaking 6 1-month-old S. grosvenorii cut seedlings with wounded root tips in a fungal conidial suspension (1 × 106 conidium/mL) for 1 h (Choi et al. 2022). Six additional seedlings were treated with sterile distilled water as controls. All the seedlings were kept in a greenhouse at 25°C with 80% humidity. After 3 weeks, the inoculated plant leaves turned yellow and dried off. After 4 weeks, all the treated plants were dead with vascular discoloration in the roots and stems. V. dahliae was re-isolated from the symptomatic stem tissues and confirmed by morphology and DNA sequencing (ITS, ACT and GPD sequences). The control plants remained healthy and no V. dahliae was isolated from them, thereby fulfilling the Koch postulate. V. dahliae is an economically important pathogen with a wide host range worldwide (Lu et al. 2021). To our knowledge, this is the first report of V. dahliae causing Verticillium wilt of S. grosvenorii in China and elsewhere, thereby providing the possibility for the integrated management of Verticillium wilt on this herb.

Studying the consequences of somatic mutations in pre-malignant and cancerous tissues is challenging due to noise in single-cell transcriptome data and difficulty in identifying the clonal identity of single cells. We optimized TARGET-seq to develop TARGET-seq+, which combines RNA sequencing (RNA-seq), the analysis of cell surface protein expression, and genotyping in single cells with improved sensitivity. We describe the steps for cell isolation, the preparation of single-cell RNA-seq (scRNA-seq) and genotyping libraries, and sequencing. We also provide guidance on the analysis of single-cell genotyping, transcriptome pre-processing, and data integration. For complete details on the use and execution of this protocol, please refer to Jakobsen et al.1.

Plant post-embryonic development is characterized by the unceasing generation of new tissues and organs that allow plants to adapt their growth and architecture to environmental conditions. Primary root indeterminate growth is sustained by the activity of the root apical meristem (RAM), an organized structure containing stem cells that continuously divide to generate new tissues. RAM, thanks to its simple and symmetric structure, is an excellent system for studying how hormone distributions and interactions regulate growth. Plant hormones are pivotal actors in establishing and maintaining RAM patterning, and their developmental outputs are dependent on their integrated activities. In this review, we discuss recent findings on hormonal crosstalk that shapes the root meristem, focusing our attention on both the root radial and longitudinal axes.

LIPUS, low intensity pulsed ultrasounds, are considered a safe and non-invasive tool that have been used extensively in medicine for chronic diseases. We evaluated the effects produced by LIPUS on the physiological behaviour of mouse and human adipose stem cells (ASCs) as well as on adipose tissue explants. LIPUS stimulation for one minute did not affect mouse or human ASC proliferation or apoptosis, with no evident changes in morphology or cell growth. Further, RNA-seq analysis revealed more than 200 genes differentially expressed after ultrasound stimulation of mouse and human ASCs. Among them, the changes in gene expression mainly belong to the canonical pathways of stemness, energy metabolism or chemokine signalling. On the other hand, the migration ability of mouse and human ASCs was partially affected by the ultrasound protocol, slightly increasing their capacity to migrate, while maintaining their stemness properties. Major affection was detected on the adipocyte differentiation capacity. Indeed, LIPUS-treated ASCs were unable to differentiate into mature adipocytes and their inflammatory secretion profile was reduced in both mouse and human ASCs. These effects were confirmed on an ex vivo model of adipose tissue explants, demonstrating that LIPUS also provoked an anti-inflammatory profile in the adipose tissue while reducing adipocytes differentiation and lipids content. Overall, the strong adipogenesis blockade, the reduction of their secretion profile and the RNA-seq analysis suggest that LIPUS-treated ASCs may be prevented from differentiating into mature adipocytes and from exacerbating inflammation, thus limiting their contribution to obesity.

The functionality and structural integrity of the cardiovascular system are critically dependent on vascular smooth muscle cells (VSMCs). Human mesenchymal stem cells (hMSCs) have significant potential for differentiating into VSMCs, making them a valuable resource in regenerative medicine and the development of vascular grafts. This study explored the synergistic effects of micropatterned substrates and TGF-β1 on the differentiation of hMSCs into VSMCs. HMSCs were cultured on both micropatterned and flat substrates for a duration of 6 days, with some groups receiving TGF-β1 treatment, after which cell morphology and the expression of specific smooth muscle markers were evaluated through Western blotting, immunofluorescence staining, and RT-qPCR. Results indicated that hMSCs on micropatterned substrates treated with TGF-β1 exhibited significantly elevated protein levels of smooth muscle myosin heavy chain (MYH11) compared with hMSCs on flat substrates without TGF-β1 (p < 0.001). Additionally, MYH11 expression was markedly enhanced in samples cultured on micropatterned substrates with TGF-β1. Furthermore, hMSCs treated with TGF-β1 on flat substrates exhibited increased cadherin-11 mRNA expression compared with both micropatterned and flat substrates lacking TGF-β1 (p < 0.05). Interestingly, KLF4 protein levels were significantly higher in hMSCs on flat substrates without TGF-β1 compared to those cultured on micropatterned substrates with TGF-β1 treatment (p < 0.001). In conclusion, this study demonstrated that the combination of micropatterned substrates and TGF-β1 treatment preferentially enhances MYH11 expression, indicative of advanced smooth muscle cell organization, along with modulating KLF4 levels and upregulating cadherin-11 expression in hMSCs. These findings provide critical insights into the differentiation pathways of MSCs into VSMCs and may inform the design of improved vascular grafts that better replicate the properties of native blood vessels.

Mixed chimerism occurs frequently with the risk of graft rejection for aplastic anemia patients undergoing matched sibling donor hematopoietic stem cell transplantation in cyclophosphamide (CY) and anti-thymocyte globulin (ATG) conditioning. So far, no one knows how to adjust immunosuppression (IS) during MC. We retrospectively analyzed 87 consecutive pediatric patients. Early withdrawal (EW) IS and donor lymphocyte infusion were attempted to reverse MC. The rate of MC was 26% (n = 23). Low dose CY (120-150 mg/kg) was a risk factor for MC (P = 0.0002) and increasing the dosage of fludarabine did not eliminate it. Patients receiving 200 mg/kg CY had the lowest MC rate (8%) and best 3-year graft-versus-host disease/failure-free survival (GFFS; 95%). Donor chimerism in T cells was more sensitive than that in whole blood (P = 0.001). In 17 patients with early-onset MC, EW IS strategy was helpful in improving complete chimerism (CC) (EW cohort: 63 versus non-EW cohort: 295 days; P = 0.008). Our study shows that CY + ATG conditioning needs to be intensified to maintain engraftment and 200 mg/kg CY + 150 mg/m2 FLU is recommended for basic conditioning. The EW IS strategy should be considered as an important option to improve donor chimerism in early-onset MC. Clinical trial registration: URL: https://www.chictr.org.cn ; ChiCTR-1900023509. (Retrospective registration in 2019/5/31).

This review explores the role of cell communication network (CCN) proteins in regulating skeletal physiology, aging, and disease, particularly within the context of balanced bone remodeling.

To perform a systematic review and meta-analysis of randomized and comparative studies comparing mesenchymal stromal cells other orthobiological injections for patients with knee osteoarthritis.

Outer retinal function depends on two supporting tissues: the retinal pigment epithelium (RPE) and the choroid. Limited molecular information is available on the intercellular networks that sustain RPE/choroid tissue in both healthy and pathological states. Galectin-1 (Gal1), a β-galactoside-binding lectin, has recently emerged as a key regulator of angiogenesis and a potential therapeutic target in vascular pathologies, including age-related macular degeneration. Here, we studied the expression of Gal1 in the outer retina and its regulatory role in the RPE/choroid under physiological and pathological conditions. Our findings indicate that Gal1 is predominantly associated with stromal cells in the RPE/choroid. In Gal1-deficient (Lgals1-/-) mice, the RPE/choroid ultrastructure and gene expression profiles were altered, and choroidal explants exhibited reduced sprouting compared to those of wild-type mice. Consistently, recombinant Gal1 promoted choroidal sprouting under hypoxic conditions, and stromal-like cells modulated pro-angiogenic and antiangiogenic gene expression in vitro under pathological conditions. Interestingly, Gal1 was also expressed by the RPE, with apical secretion under normoxia that shifted toward a basolateral phenotype under hypoxia. These findings identify stromal-like cells and RPE as key sources of Gal1 in the choroid, highlighting its distinct roles in maintaining RPE/choroid homeostasis in healthy or pathological microenvironments.

Vascular repair and regeneration are critical for tissue homeostasis. Endothelial colony forming cells (ECFCs) are vessel-resident progenitors with vasoreparative capacity and they offer an important avenue for allogeneic cytotherapy to achieve perfusion of ischemic tissues. Endothelial Protein C Receptor (EPCR) has been proposed as a marker for vascular endothelial stem cells, but its precise role in ECFC biology remains unknown. The current study has investigated the biological relevance of EPCR in ECFC function. Our data show that over 95% of ECFCs exhibit high EPCR expression. These levels surpassing CD34 and CD157, positions EPCR as a new robust ECFC immunophenotypic marker, alongside established markers CD31 and CD105. Functionally, depleting EPCR expression in ECFCs significantly diminished angiogenic activity, including proliferation, migration and tube formation. This knockdown also altered normal ECFC barrier function. Transcriptomic analysis indicated that knockdown of EPCR led to enrichment of gene signatures for cell cycle, TGF beta, and focal adhesion kinases. G1 cell cycle arrest was confirmed in ECFCs with depleted EPCR. Mechanistically, EPCR knockdown led to increased release of TGFβ2 and SMAD2/3 activation, coupled with increased p21, decreased pFAK, and increased transgelin. Additionally, we showed that quiescent ECFCs showed significantly lower EPCR expression when compared to proliferating ECFCs. In agreement with this, cell sorting experiments demonstrated that ECFCs with the highest EPCR expression exhibited the highest clonogenic capacity. In summary, our findings highlight that EPCR expression in ECFCs is critical for their angiogenic activity, by modulating cell cycle progression.

FGM is an issue of increasing concern also in countries where it is not traditionally practiced. Vulvar scarring is the most common long-term effect associated with FGM, representing one of the main unmet issues in FGM women's health. Regenerative therapies based on the use of adipose-derived stem cells are considered the standard of care for ameliorating scarring and fibrosis. This study aimed to explore the potential of fat grafting in the treatment of post-FGM vulvar scars.

Selenium (Se) is a highly biologically active element, and its organic derivatives have attracted growing interest for their promising chemotherapeutic potential, largely due to their redox-modulating activity, which selectively affects cancer cells with high levels of reactive oxygen species (ROS). However, their high reactivity and susceptibility to spontaneous degradation limit their biomedical application. To harness their potential in the realm of nanomedicine, we present a new generation of therapeutically promising polymers that combine Se with 2,2-bis(methylol)propionic acid (bis-MPA)-based dendritic polymers, chosen for their high chemical versatility, low toxicity, and excellent biodegradability. Most examples in the literature about dendritic polymers feature dormant dendritic skeletons with active functional groups expressed only on their periphery, which severely limits their functional scope. In this work, monodisperse dendrimers and linear-dendritic (LD) polymers up to the third generation were developed, with the latter capable of self-assembling into dendritic micelles (∼20 nm). These systems feature Se at the dendritic core or peripheral branches in the form of monoselenide or diselenide bridges. Selenium incorporation demonstrated excellent compatibility with two key polyester synthetic approaches: anhydride chemistry and fluoride-promoted esterification (FPE). Both monoselenide and diselenide linkages introduced degradability and dynamic behavior in dendrimers and dendritic micelles. However, their biological activities differed significantly. Diselenide-containing dendrimers exhibited great anticancer potential against breast cancer cell lines, with IC50 values in the micromolar range. Among these, first-generation Se dendrimers stood out due to their promising selectivity toward cancer cells. In contrast, dendritic polymers incorporating monoselenides retained the high biocompatibility characteristics of bis-MPA dendritic constructs.