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Chronic tympanic membrane (TM) perforation is a tubotympanic disease caused by either traumatic injury or inflammation. A recent study demonstrated significant progress in promoting the regeneration of chronic TM perforations through the application of nanofibers with radially aligned nanostructures and controlled release of growth factors. However, radially aligned nanostructures with stem cell-stimulating factors have never been used. In this study, insulin-like growth factor binding factor 2 (IGFBP2)-incorporated radially aligned nanofibrous patches (IRA-NFPs) were developed and applied to regenerate chronic TM perforations. The IRA-NFPs were prepared by electrospinning 8 wt% polycaprolactone in trifluoroethanol and acetic acid (9:1). Random nanofibers (RFs) and aligned nanofibers (AFs) were successfully fabricated using a flat plate and a custom-designed circular collector, respectively. The presence of IGFBP2 was confirmed via Fourier transform infrared spectroscopy and the release of IGFBP2 was sustained for up to 20 days. In vitro studies revealed enhanced cellular proliferation and migration on AFs compared to RFs, and the incorporation of IGFBP2 further promoted these effects. Quantitative real-time PCR revealed mRNA downregulation, correlating with accelerated migration and increased cell confluency. In vivo studies showed IGFBP2-loaded RF and AF patches increased regeneration success rates by 1.59-fold and 2.23-fold, respectively, while also reducing healing time by 2.5-fold compared to the control. Furthermore, IGFBP2-incorporated AFs demonstrated superior efficacy in healing larger perforations with enhanced histological similarity to native TMs. This study, combining stem cell stimulating factors and aligned nanostructures, proposes a novel approach potentially replacing conventional surgical methods for chronic TM perforation regeneration. Chronic otitis media (COM) affects approximately 200 million people worldwide due to inflammation, inadequate blood supply, and lack of growth factors. Current surgical treatments have limitations like high costs and anesthetic risks. Recent research explored the use of nanofibers with radially aligned nanostructures and controlled release of growth factors to treat chronic tympanic membrane (TM) perforations. In this study, insulin-like growth factor binding protein 2 (IGFBP2)-incorporated radially aligned nanofibrous patches (IRA-NFPs) were developed and applied to regenerate chronic TM perforations. We assessed their properties and efficacy through in vitro and in vivo studies. IRA-NFPs showed promising healing capabilities with chronic TM perforation models. This innovative approach has the potential to improve COM management, reduce surgery costs, and enhance patient safety. [Display omitted] [ABSTRACT FROM AUTHOR]

Stem cell-based therapies offer promising avenues for treating inflammatory diseases owing to their immunomodulatory properties. However, challenges persist regarding their survival and efficacy in inflamed tissues. Our study introduces a novel approach by engineering adipose-derived stem cells (ADSCs) to enhance their viability in inflammatory environments and boost the secretion of paracrine factors for treating inflammatory bowel disease (IBD). An arginine-glycine-aspartate peptide-poly (ethylene glycol)-chlorin e6 conjugate (RPC) was synthesized and coupled with ADSCs, resulting in RPC-labeled ADSCs (ARPC). This conjugation strategy employed RGD-integrin interaction to shield stem cells and allowed visualization and tracking using chlorin e6. The engineered ARPC demonstrated enhanced viability and secretion of paracrine factors upon light irradiation, regulating the inflammatory microenvironment. RNA-sequencing analysis unveiled pathways favoring angiogenesis, DNA repair, and exosome secretion in ARPC(+) while downregulating inflammatory pathways. In in vivo models of acute and chronic IBD, ARPC(+) treatment led to reduced inflammation, preserved colon structure, and increased populations of regulatory T cells, highlighting its therapeutic potential. ARPC(+) selectively homed to inflammatory sites, demonstrating its targeted effect. Overall, ARPC(+) exhibits promise as an effective and safe therapeutic strategy for managing inflammatory diseases like IBD by modulating immune responses and creating an anti-inflammatory microenvironment. [Display omitted] [ABSTRACT FROM AUTHOR]

Recently, the formation of three-dimensional (3D) cell aggregates known as embryoid bodies (EBs) grown in media supplemented with HSC-specific morphogens has been utilized for the directed differentiation of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), into clinically relevant hematopoietic stem cells (HSCs). However, delivering growth factors and nutrients have become ineffective in inducing synchronous differentiation of cells due to their 3D conformation. Moreover, irregularly sized EBs often lead to the formation of necrotic cores in larger EBs, impairing differentiation. Here, we developed two gelatin microparticles (GelMPs) with different release patterns and two HSC-related growth factors conjugated to them. Slow and fast releasing GelMPs were conjugated with bone morphogenic factor-4 (BMP-4) and stem cell factor (SCF), respectively. The sequential presentation of BMP-4 and SCF in GelMPs resulted in efficient and effective hematopoietic differentiation, shown by the enhanced gene and protein expression of several mesoderm and HSC-related markers, and the increased concentration of released HSC-related cytokines. In the present study, we were able to generate CD34+, CD133+, and FLT3+ cells with similar cellular and molecular morphology as the naïve HSCs that can produce colony units of different blood cells, in vitro. [Display omitted] • Successful conjugation of BMP4 and SCF in fast and slow release gelatin microparticles. • BMP4/SCF conjugation in GelMPs promotes proliferation and reduces apoptosis. • Dual GelMP system promotes sequential differentiation of iPSC to mesoderm and HSC. • CD34+ iPS-derived HSCs (iHSCs) are capable of producing various blood cells, in-vitro. • Differentiated iHSCs were verified by RNA sequencing analysis. [ABSTRACT FROM AUTHOR]

Vitiligo is a skin disorder with melanocyte destruction caused by complex interplay between multiple genetic and environmental factors. Recent studies have suggested DNA methylation is involved in the melanocyte damage, but the underlying mechanism remains unknown. To explore the abnormal DNA methylation patterns in vitiligo lesional and nonlesional skin, and the mechanism of DNA methylation involved in vitiligo pathogenesis. Initially, the genome-wide aberrant DNA methylation profiles in lesional and nonlesional skin of vitiligo were detect via Illumina methylation EPIC 850k Beadchip. Subsequently, a comprehensive analysis was conduct to investigate the genomic characteristics of differentially methylated regions (DMRs). Furthermore, the effects of key aberrant methylated genes on cell apoptosis and function of both melanocytes and keratinocytes were further identified and validated by western bloting, ELISA, and immunofluorescence. Compared with nonlesional skins, we discovered 79 significantly differentially methylated CpG sites in vitiligo lesions. These DMRs were mainly located in the gene body and the TS1500 region. Annexin A2 receptor (ANXA2R), a crucial gene in cell apoptosis, was hypermethylated in vitiligo lesions. Furthermore, we showed that ANXA2R displayed hypermethylation and low expression levels in both keratinocytes and melanocytes of vitiligo patients, and the hypermethylated-triggered downregulation of ANXA2R under oxidative stress induced melanocyte apoptosis, and inhibited the secretion of stem cell factor (SCF) from keratinocytes thus impaired the survival of melanocytes. Our study illustrates the DNA methylation modification in vitiligo, and further demonstrates the molecular mechanism of hypermethylated ANXA2R in the dysfunction of melanocytes under oxidative stress. • Our study provides a map of abnormal DNA methylation in vitiligo lesions, and clarifying its role in disease development. • Clarifying the role of hypermethylated gene ANXA2R in vitiligo by inducing melanocyte apoptosis. • Revealing hypermethylated ANXA2R in vitiligo inhibiting the secretion of stem cell factor from keratinocytes thus impairing melanocyte survival. [ABSTRACT FROM AUTHOR]

Constipation is directly related to the intestinal microenvironment, in which the promotion of gastrointestinal (GI) motility and improvement of gut microbiota distribution are important for alleviating symptoms. Herein, after the intervention of probiotic fermented milk (FMMIX) containing Lacticaseibacillus paracasei JY062 and Lactobacillus gasseri JM1 for 14 d in Kunming mice with loperamide-induced constipation, the results indicated that FMMIX significantly increased the secretion of serum motilin, gastrin and 5-hydroxytryptamine, as well as decreased the secretion of peptide YY, vasoactive intestinal peptide, and nitric oxide in mice. As determined by immunohistochemical analysis, FMMIX promoted an augmentation in the quantity of Cajal interstitial cells. In addition, the mRNA and protein expression of c-kit and stem cell factor (SCF) were upregulated to facilitate intestinal motility. High-throughput sequencing and gas chromatography techniques revealed that FMMIX led to an increase in the relative abundance of beneficial bacteria (Lactobacillus , Oscillospira , Ruminococcus , Coprococcus , and Akkermansia), reduced the presence of harmful bacteria (Prevotella), and resulted in elevated levels of short-chain fatty acids (SCFA) with a superior improvement compared with unfermented milk. Untargeted metabolomics revealed significant upregulation of functional metabolites such as l -pipecolinic acid, dl -phenylalanine, and naringenin in FMMIX, presumably playing a potential role in constipation relief. Overall, our results showed that FMMIX had the potential to alleviate constipation symptoms in mice by improving the secretion of serum GI regulatory peptides and neurotransmitters, increasing the expression of c-kit and SCF proteins, and modulating the gut microbiota structure and SCFA levels, and may be associated with an increase in these functional metabolites. This suggested that FMMIX could be a promising adjunctive strategy for managing constipation symptoms and could contribute to the development of functional foods aimed at improving gut health. [Display omitted] [ABSTRACT FROM AUTHOR]

Citric acid (CA) conditioning may be a promising alternative to ethylenediaminetetraacetic acid (EDTA) in regenerative endodontic procedures, as reported to improve growth factors' release from dentin. This review systematically investigated the effect of CA conditioning on the growth factors release from dentin and cell behavior compared to EDTA conditioning. Searches were conducted (PubMed/MEDLINE, Scopus, Web of Science, Embase, SciELO, Cochrane Library, and grey literature) until May-2023. Only in vitro studies that evaluated the effects of CA on growth factors' release from dentin and cell behavior outcomes compared to EDTA were included. The studies were critically appraised using a modified Joanna Briggs Institute's checklist. Meta-analysis was unfeasible. Out of the 335 articles screened, nine were included. Among these, three studies used dentin discs/roots from permanent human teeth; the rest combined them with stem cells. 10% CA for 5 or 10 minute was the most used protocol. Meanwhile, EDTA concentrations ranged from 10% to 17%. In eight studies examining the release of growth factors, five reported a significant release of transforming growth factor-β after dentin conditioning with 10% CA compared to 17% EDTA. Regarding cell behavior (6 studies), three studies assessed cell viability. The findings revealed that 10% CA conditioning showed cell viability similar to those of 17% EDTA. Additionally, in two out of three studies, it was observed that 10% CA conditioning did not affect cell morphology. The studies had a low risk of bias. The use of 10% CA to condition dentin for 5-10 minutes resulted in a notable transforming growth factor -β1 release, but its cell responses were similar to those of EDTA. [ABSTRACT FROM AUTHOR]

• Samd14 deletion caused more severe anemia after 5-fluorouracil-induced stress • Mast cell differentiation was attenuated in Samd14 knockout mouse model (Samd14-CKO) cells • Samd14-CKO mast cells were functionally impaired and less reactive to stem cell factor (SCF) and IL-3 • Samd14-CKO mice were similar to other Kit loss-of-function mouse models Sterile α-motif domain-14 (Samd14) protein expression increases the regenerative capacity of the erythroid system. Samd14 is transcriptionally upregulated and promotes cell signaling via the receptor tyrosine kinase Kit in a critical window of acute erythroid regeneration. We generated a hematopoietic-specific conditional Samd14 knockout mouse model (Samd14-CKO) to study the role of Samd14 in hematopoiesis. The Samd14-CKO mouse was viable and exhibited no steady-state hematopoietic phenotype. Samd14-CKO mice were hypersensitive to 5-fluorouracil, resulting in more severe anemia during recovery and impaired erythroid progenitor colony formation. Ex vivo, Samd14-CKO hematopoietic progenitors were defective in their ability to form mast cells. Samd14-CKO mast cells exhibited altered Kit/stem cell factor (SCF), IL-3/IL-3R signaling, and less granularity than Samd14-FL/FL cells. Our findings indicate that Samd14 promotes both erythroid and mast cell functions. The Samd14-CKO mouse phenotype exhibits striking similarities to the KitW/W-v mice, which carry Kit mutations resulting in reduced tyrosine kinase–dependent signaling, causing mast cell and erythroid abnormalities. The Samd14-CKO mouse model is a new tool for studying hematologic pathologies involving Kit signaling. [ABSTRACT FROM AUTHOR]

• COVID-19 vaccination post-COVID-19 conditions (PCC) reduced the number of symptoms and increased well-being. • COVID-19 vaccination post-PCC down-regulated systemic markers of inflammation. • COVID-19 vaccination post-PCC did not abrogate the persistence of viral products. • A viral reservoir not cleared by one or two vaccine doses may persist. Vaccination protects against severe COVID-19 manifestations. For those with post-COVID-19 conditions (PCC) or long COVID, the impact of COVID-19 vaccination on the evolution of symptoms, immune responses, and viral persistence is unclear. In this prospective observational cohort study, we evaluated the number of PCC symptoms, affected organ systems, and psychological well-being scores before and after patients with PCC received COVID-19 vaccination. We simultaneously evaluated biomarkers of systemic inflammation and levels of plasma cytokines/chemokines. We measured plasma and intracellular levels of SARS-CoV-2 antigens, and immunoreactivity to SARS-CoV-2 antigens in blood. COVID-19 vaccination was associated with decreases in number of PCC symptoms (pre-vaccination: 6.56 ± 3.1 vs post-vaccination: 3.92 ± 4.02; P < 0.001) and affected organ systems (pre-vaccination: 3.19 ± 1.04 vs post-vaccination: 1.89 ± 1.12; P < 0.001), and increases in World Health Organization (WHO)-5 Well-Being Index Scores (pre-vaccination: 42.67 ± 22.76 vs post-vaccination: 56.15 ± 22.83; P < 0.001). Patients with PCC also had significantly decreased levels of several pro-inflammatory plasma cytokines/chemokines after COVID-19 vaccination including sCD40L, GRO-⍺, macrophage inflammatory protein (MIP)-1⍺, interleukin (IL)-12p40, G-colony stimulating factor (CSF), M-CSF, IL-1β, and stem cell factor (SCF). PCC participants presented a certain level of immunoreactivity toward SARS-CoV-2, that was boosted with vaccination. SARS-CoV-2 S1 antigen persisted in the blood of PCC participants, mostly in non-classical monocytes, regardless of participants receiving vaccination. Our study shows higher pro-inflammatory responses associated with PCC symptoms and brings forward a possible role for vaccination in mitigating PCC symptoms by decreasing systemic inflammation. We also observed persistence of viral products independent of vaccination that could be involved in perpetuating inflammation through non-classical monocytes. [ABSTRACT FROM AUTHOR]

Herein, we present an approach for manipulating paracrine factors and signaling pathways in adipose-derived stem cells (ADSCs) to achieve highly effective tumor immunotherapy. Our method involves precise control of reactive oxygen species concentration using the CD90-maleimide-pluronic F68-chlorin e6 conjugate (CPFC) to create ACPFC, which is then attached to ADSCs through the CD90 receptor-specific interaction. By regulating the irradiated laser power, ACPFC promotes signaling pathways such as cascade-3, VEGFR2, α2β1, C3AR1, CR1–4, and C5AR1, leading to the secretion of various inflammatory cytokines such as IFN-γ, TGF-β, and IL-6, while inhibiting AKT, ERK, NFkB, PAR1, and PAR3/4 signaling pathways to reduce the secretion of cell growth factors like TIMP-1, TIMP-2, VEGF, Ang-2, FGF-2, and HGF. When ACPFC is injected intravenously into a tumor animal model, it autonomously targets and accumulates at the tumor site, and upon laser irradiation, it generates various anti-inflammatory factors while reducing angiogenesis growth factors. The resulting antitumor response recruits CD3+CD8+ cytotoxic T cells and CD3+CD4+ helper T cells into the tumor and spleen, leading to highly effective melanoma and pancreatic tumor treatment in mice. Our technology for regulating stem cell paracrine factors holds significant promise for the treatment of various diseases. [Display omitted] • ROS mediates regulation of stem cell paracrine factors. • Targeted ROS manipulation via CPFC binding to ADSCs. • Effective tumor homing and accumulation by ACPFC. • Tumor growth inhibition and immune activation in vivo. • Insights into paracrine factors in stem cell-based immunotherapy. [ABSTRACT FROM AUTHOR]

Oligoasthenospermia is the primary cause of infertility. However, there are still enormous challenges in the screening of critical candidates and targets of oligoasthenospermia owing to its complex mechanism. In this study, stem cell factor (SCF), c-kit, and transient receptor potential vanilloid 1 (TRPV1) biosensors were successfully established and applied to studying apoptosis and autophagy mechanisms. Interestingly, the detection limit reached 2.787 × 10−15 g/L, and the quantitative limit reached 1.0 × 10−13 g/L. Furthermore, biosensors were used to investigate the interplay between autophagy and apoptosis. Schisandrin A is an excellent candidate to form a system with c-kit similar to SCF/c-kit with a detection constant (K D) of 5.701 × 10−11 mol/L, whereas it had no affinity for SCF. In addition, it also inhibited autophagy in oligoasthenospermia through antagonizing TRPV1 with a K D of up to 4.181 × 10−10 mol/L. In addition, in vivo and in vitro experiments were highly consistent with the biosensor. In summary, high-potency schisandrin A and two potential targets were identified, through which schisandrin A could reverse the apoptosis caused by excessive autophagy during oligoasthenospermia. Our study provides promising insights into the discovery of effective compounds and potential targets via a well-established in vitro - in vivo strategy. Discovery of Schisandrin A regulating the interplay of autophagy and apoptosis of oligoasthenospermia by targeting the SCF/c-kit and TRPV1 via biosensors coupled with in vivo and in vitro investigation. [Display omitted] [ABSTRACT FROM AUTHOR]

Mast cells are essential regulators of inflammation most recognized for their central role in allergic inflammatory disorders. Signaling via the high-affinity immunoglobulin E (IgE) receptor, FceRI, leads to rapid degranulation of preformed granules and the sustained release of newly synthesized proinflammatory mediators. Our group recently established rosemary extract as a potent regulator of mast cell functions, attenuating MAPK and NF-κB signaling. Carnosic acid (CA)--a major polyphenolic constituent of rosemary extract--has been shown to exhibit anti-inflammatory effects in other immune cell models, but its role as a potential modulator of mast cell activation is undefined. Therefore, we sought here to determine the modulatory effects of CA in a mast cell model of allergic inflammation. We sensitized bone marrow-derived mast cells with anti-trinitrophenyl IgE and activated with allergen (TNP-BSA) under stem cell factor potentiation, in addition to treatment with CA. Our results indicate that CA significantly inhibits allergen-induced early phase responses including Ca2+ mobilization, ROS production, and subsequent degranulation. We also show CA treatment reduced late phase responses, including the release of all cytokines and chemokines examined following IgE stimulation and corresponding gene expression excepting that of CCL2. Importantly, we determined that CA mediates its inhibitory effects through modulation of tyrosine kinase Syk and downstream effectors TAK1 (Ser412) and Akt (Ser473) as well as NFκB signaling, while phosphorylation of FceRI (γ chain) and MAPK proteins remained unaltered. These novel findings establish CA as a potent modulator of mast cell activation, warranting further investigation as a putative anti-allergy therapeutic. [ABSTRACT FROM AUTHOR]

Fibroblast growth factor (FGF) is a multifunctional protein that exhibits a wide range of biological effects. Most commonly, it acts as a mitogen, but it also has regulatory, morphological, and endocrine effects. The four receptor subtypes of FGF are activated by more than 20 different FGF ligands. FGF2, one of the FGF ligands, is an essential factor for cell culture in stem cells for regenerative medicine; however, recombinant FGF2 is extremely unstable. Here, we successfully generated homobivalent agonistic single-domain antibodies (variable domain of heavy chain of heavy chain antibodies referred to as VHHs) that bind to domain III and induce activation of the FGF receptor 1 and thus transduce intracellular signaling. This agonistic VHH has similar biological activity (EC50) as the natural FGF2 ligand. Furthermore, we determined that the agonistic VHH could support the proliferation of human-induced pluripotent stem cells (PSCs) and human mesenchymal stem cells, which are PSCs for regenerative medicine. In addition, the agonistic VHH could maintain the ability of mesenchymal stem cells to differentiate into adipocytes or osteocytes, indicating that it could maintain the properties of PSCs. These results suggest that the VHH agonist may function as an FGF2 mimetic in cell preparation of stem cells for regenerative medicine with better cost effectiveness. [ABSTRACT FROM AUTHOR]

c-KIT and its ligand stem cell factor (SCF) play a direct role in the oncogenesis of various cancers by regulating the cell fate. Recent evidence indicates that an increased expression of c-KIT/SCF, driven by hormonal imbalances, is an important step in the development of hormone-dependent cancers. We investigated the possible role of the c-KIT/SCF system in the carcinogenesis in 44 perianal gland tumours (16 adenomas, 15 epitheliomas and 13 carcinomas) and 10 normal perianal gland tissues by assessing the percentage and type of cells that expressed c-KIT and SCF as well as the cellular localization of immunoreactivity. No differences in immunolabelling of SCF were found between normal glands and neoplastic cells of any histotype. The highest expression of c-KIT was seen in carcinomas and a positive correlation was found between c-KIT labelling score and mitotic index (R = 0.876; P <0.01). c-KIT labelling patterns in hepatoid cells varied among the tumour histotypes with adenomas having only membranous labelling. Three labelling patterns (membranous only, membranous and cytosolic, and cytosolic only) were seen in the other tumour histotypes. Cytosolic labelling was statistically more frequent in carcinomas than in adenomas (P <0.001). These findings suggest that c-KIT expression and its cellular localization may play a role in the development and progression of perianal gland tumours by influencing cell proliferation. [ABSTRACT FROM AUTHOR]

Many mediators are produced during pulp inflammation and necrosis, including endocannabinoids (ECbs), which might affect the function of stem cells of the apical papilla (SCAP), cells of paramount importance for root formation, and regenerative endodontic treatment. The aim of this study was to evaluate the production of osteoclastogenesis-related mediators by SCAP modulated by ECbs and lipopolysaccharide (LPS) in vitro. SCAP were cultured and treated with ECb anandamide (AEA), 2-arachidonoylglycerol, or N-arachidonoylaminophenol. All groups were incubated in the presence of a vehicle or LPS and the antagonist of transient receptor potential cation channel subfamily V member 1, capsazepine. After 24 hours, the culture medium supernatants were collected for further quantification of tumor necrosis factor alpha, CCL2, macrophage colony-stimulating factor, osteoprotegerin, and receptor activator of nuclear factor kappa B ligand. Small amounts of tumor necrosis factor alpha and receptor activator of nuclear factor kappa B ligand were detected in SCAP supernatants, and none of the experimental conditions altered their production. A down-regulation in constitutive CCL2 production was observed in the AEA group compared with that in the LPS group. The production of macrophage colony-stimulating factor was significantly increased in all groups treated with AEA compared with the control and LPS-treated groups. Osteoprotegerin was significantly increased by AEA alone and by 2-arachidonoylglycerol and N-arachidonoylaminophenol in the presence of LPS and capsazepine. AEA modulates some of the osteoclastogenic factors produced by SCAP in a bone resorption protective fashion. [ABSTRACT FROM AUTHOR]

Microfracture treatment that is basically relied on stem cells and growth factors in bone marrow has achieved a certain progress for cartilage repair in clinic. Nevertheless, the neocartilage generated from the microfracture strategy is limited endogenous regeneration and prone to fibrosis due to the influences of cell inflammation and vascular infiltration. To explore the crucial factor for articular cartilage remodeling, here we design a trilaminar osteochondral scaffold with a selective permeable film in middle isolation layer which can prevent stem cells, immune cells, and blood vessels in the bone marrow from invading into the cartilage layer, but allow the nutrients and cytokines to penetrate. Our findings show that the trilaminar scaffold exhibits a good biocompatibility and inflammatory regulation, but the osteochondral repair is far less effective than the control of double-layer scaffold without isolation layer. These results demonstrate that it is not adequate to rely only on nutrients and cytokines to promote reconstruction of articular cartilage, and the various cells in bone marrow are indispensable. Consequently, the current study illustrates that cell infiltration involving stem cells, immune cells and other cells from bone marrow plays a crucial role in articular cartilage remodeling based on the integrated scaffold strategy. Clinical microfracture treatment plays a certain role on the restoration of injured cartilage, but the regenerative cartilage is prone to be fibrocartilage due to the modulation of bone marrow cells. Herein, we design a trilaminar osteochondral scaffold with a selective permeable film in middle isolation layer. This specific film made of dense electrospun nanofiber can prevent bone marrow cells from invading into the cartilage layer, but allow the nutrients and cytokines to penetrate. Our conclusion is that the cartilage remodeling will be extremely inhibited when the bone marrow cells are blocking. Owing to the diverse cells in bone marrow, we will further explore the influence of each cell type on cartilage repair in our continuous future work. [Display omitted] [ABSTRACT FROM AUTHOR]

Hydrogels can possess desired biochemical and mechanical properties, excellent biocompatibility, satisfactory biodegradability, and biological capabilities that promote skin repair, making them ideal candidates for skin healing dressings. Polysaccharides, such as chitosan, hyaluronic acid and sodium alginate as well as proteins, including gelatin, collagen and fibroin proteins, are biological macromolecules celebrated for their biocompatibility and biodegradability, are at the forefront of innovative hydrogel dressing development. This work first summarizes the skin wound healing process and its influencing factors, and then systematically articulates the multifunctional roles of hydrogels based on biological macromolecules (polysaccharides and proteins) as dressing in addressing bacterial infection, hemorrhage and inflammation during wound healing. Furthermore, this review explores the potential of these hydrogels as vehicles for combination therapy, by incorporating growth factors or stem cells. Finally, the article offers insights into future directions of such hydrogels in wound repair field. [Display omitted] [ABSTRACT FROM AUTHOR]

The placenta is essential organ for oxygen and nutrient exchange between the mother and the developing fetus. Trophoblast lineage differentiation is closely related to the normal function of the placenta. Trophoblast stem cells (TSCs) can differentiate into all placental trophoblast subtypes and are widely used as in vitro stem cell models to study placental development and trophoblast lineage differentiation. Although extensive research has been conducted on the differentiation of TSCs, the possible parallels between trophoblast giant cells (TGCs) that are differentiated from TSCs in vitro and the various subtypes of TGC lineages in vivo are still poorly understood. In this study, mouse TSCs (mTSCs) were induced to differentiate into TGCs, and our mRNA sequencing (RNA-seq) data revealed that mTSCs and TGCs have distinct transcriptional signatures. We conducted a comparison of mTSCs and TGCs transcriptomes with the published transcriptomes of TGC lineages in murine placenta detected by single-cell RNA-seq and found that mTSCs tend to differentiate into maternal blood vessel-associated TGCs in vitro. Moreover, we identified the transcription factor (TF) ZMAT1, which may be responsible for the differentiation of mTSCs into sinusoid TGCs, and the TFs EGR1 and MITF, which are likely involved in the differentiation of mTSCs into spiral artery-associated TGCs. Thus, our findings provide a valuable resource for the mechanisms of trophoblast lineage differentiation and placental deficiency-associated diseases development. • mTSCs and TGCs have distinct transcriptional signatures. • mTSCs tend to differentiate into maternal blood vessel-associated TGCs in vitro. • ZMAT1 might be responsible for the differentiation of mTSCs into sinusoid TGCs. • EGR1 and MITF might be involved in the differentiation of mTSCs into spiral artery-associated TGCs. [ABSTRACT FROM AUTHOR]

Skin acts as a protective barrier for the underlying organs against external events such as irradiation of ultraviolet rays, incursion of harmful pathogens, and water evaporation. As the skin is constantly liable to damage, the wound-healing process is vital to the survival of all organisms. Materials design and development for enhanced wound healing and skin tissue regeneration have been found highly valuable in recent years. A wide range of materials and structures, including dressings and tissue-engineered substitutes composed of synthetic and/or natural biopolymers and their composites have been developed and examined. Although some have clinically been proven and are available in the market, mimicking the architecture of native extracellular matrix is still an open challenge with fundamental limitations in reproducing skin appendages, sufficient vascularization, adherence to the wound bed, and scarless wound management. Biomimetic nanofibers with tunable morphological, biological, and physicochemical features are promising candidates to overcome these drawbacks. Combined with advanced biomanufacturing and cell culturing techniques, enabling the incorporation of growth factors and stem cells within morphologically-controlled nanostructures, the fibrous structures allow the regeneration of functional skin. This paper overviews the advances in state-of-the-art strategies for designing biomimetic nanofibrous materials with a high potential for wound healing and skin regeneration. An emphasis is given to multifunctional nanocomposites with mechanobiological properties matching those of natural skin. Opportunities, challenges, and commercial status of these materials for skin repair are outlined, and their future perspective is demonstrated. The advances in smart wound management are also discussed, particularly by highlighting the potential of stimuli-responsive materials and integrated sensors in the progress of next-generation dressings for simultaneous monitoring and on-demand treatment of wounds. [ABSTRACT FROM AUTHOR]

• Stem cell therapy led to a significant improvement in the behavioral score compared to the control group. • Bone marrow stem cells may have superior therapeutic effects than those from umbilical cord blood and adipose tissue. • Stem cells undergoing neural differentiation in vitro demonstrate a more potent therapeutic effect. • The use of Matrigel in combination with stem cells is recommended to enhance the therapeutic outcome. Spinal cord injury (SCI) is a serious medical condition. The search for an effective cure remains a persistent challenge. Current treatments, unfortunately, are unable to sufficiently improve neurological function, often leading to lifelong disability. This systematic review and meta-analysis evaluated the effectiveness of stem cell therapy for SCI using canine models. It also explored the optimal protocol for implementing stem cell therapy. A comprehensive search of studies was conducted from 2000 to October 2022. This study focused on five outcomes: motor function score, histopathology, IHC, western blot, and SEP. The results demonstrated a significant improvement in locomotion post-SCI in dogs treated with stem cell therapy. The therapy also led to an average increase of 3.15 points in the Olby score of the treated dogs compared to the control group. These findings highlights stem cell therapy's potential as a promising SCI treatment. The meta-analysis suggests that using bone marrow stem cells, undergoing neural differentiation in vitro, applying a surgical implantation or intrathecal route of administration, associating matrigel in combination with stem cells, and a waiting period of two weeks before starting treatment can enhance SCI treatment effectiveness. [ABSTRACT FROM AUTHOR]

Pentacyclic triterpenoids, including ursolic acid (UA), are bioactive compounds with multiple biological activities involving anti-inflammatory effects. However, the mode of their action on mast cells, key players in the early stages of allergic inflammation, and underlying molecular mechanisms remain enigmatic. To better understand the effect of UA on mast cell signaling, here we examined the consequences of short-term treatment of mouse bone marrow-derived mast cells with UA. Using IgE-sensitized and antigen- or thapsigargin-activated cells, we found that 15 min exposure to UA inhibited high affinity IgE receptor (FcεRI)-mediated degranulation, calcium response, and extracellular calcium uptake. We also found that UA inhibited migration of mouse bone marrow-derived mast cells toward antigen but not toward prostaglandin E2 and stem cell factor. Compared to control antigen-activated cells, UA enhanced the production of tumor necrosis factor-α at the mRNA and protein levels. However, secretion of this cytokine was inhibited. Further analysis showed that UA enhanced tyrosine phosphorylation of the SYK kinase and several other proteins involved in the early stages of FcεRI signaling, even in the absence of antigen activation, but inhibited or reduced their further phosphorylation at later stages. In addition, we show that UA induced changes in the properties of detergent-resistant plasma membrane microdomains and reduced antibody-mediated clustering of the FcεRI and glycosylphosphatidylinositol-anchored protein Thy-1. Finally, UA inhibited mobility of the FcεRI and cholesterol. These combined data suggest that UA exerts its effects, at least in part, via lipid-centric plasma membrane perturbations, hence affecting the functions of the FcεRI signalosome. [ABSTRACT FROM AUTHOR]

[Display omitted] • We fabricated carbonate apatite honeycomb scaffolds with vertical uniaxial channels. • At 4 weeks post implantation, all honeycomb scaffolds augmented new bone. • The augmentation happened close to the top surface of the scaffold. • In the following 8 weeks, the height and amount of new bone increased. • HC300 resisted compression from fasciae at 12-weeks post implantation. • Honeycomb structure is inherently suitable for vertical bone augmentation. Cases of intractable dental implant require vertical bone augmentation; however, the predicted bone height and volume are difficult to obtain. In vertical bone augmentation, the contact surface between the scaffold and the bone is limited to the bottom face of the scaffold. Furthermore, the strength decrease caused by scaffold resorption leads to collapse of the augmented site, leading to a decrease in the bone volume and height. To promote bone ingrowth, we fabricated carbonate apatite (i.e., bone mineral) honeycomb (HC) scaffolds with uniaxial channels vertically penetrating the scaffold. Furthermore, we controlled the scaffold resorption rate, eventually the endurability for compression, and the bone height and volume by controlling the strut thickness. The channel aperture was controlled to be 230–260 μm to promote bone ingrowth. Furthermore, the strut thicknesses of the HC scaffolds were adjusted to 100, 200, and 300 μm to control the scaffold resorption; these scaffolds were designated as HC100, HC200, and HC300, respectively. At 4 weeks post-implantation on rabbit calvarium, all scaffolds had already vertically augmented new bone close to the top surface of the scaffold. In the following 8 weeks, the height and amount of new bone in all scaffolds increased. Notably, HC300 was resorbed synchronously with new bone formation, allowing it to endure the compression from the fasciae for 12 weeks post-implantation. Furthermore, HC300 formed larger-diameter blood vessels than those of HC100 and HC200. The HC scaffolds surpassed the various combined scaffolds and growth factors or stem cells in the ability for vertical bone augmentation. Thus, the HC structure is inherently suitable for vertical bone augmentation. Notably, the HC scaffolds with 300-μm-thick struts enhanced both new bone formation and angiogenesis. This study revealed a structurally suitable design for achieving an outstanding outcome in vertical bone augmentation. [ABSTRACT FROM AUTHOR]

MTH1 protein can sanitize the damaged (d)NTP pool and MTH1 inhibitors have been developed to impede the growth of rapidly proliferating tumor cells; however, the effect of MTH1 inhibition on breast cancer stemness has not been reported yet. Here, we constructed breast cancer cell lines with the stable depletion of MTH1. MTH1 suppression clearly increased the ratio of CD44+CD24−/low subpopulations and promoted the formation of tumorspheres in MCF7 and T47D cells. RNA expression profiling, RT–qPCR and Western blotting showed the upregulation of master stem cell transcription factors Sox2, Oct4 and Nanog in MTH1 knockdown cells. GSEA suggested and Western blotting verified that MTH1 knockdown increased the expression of phosphorylated STAT3 (Tyr705). Furthermore, we indirectly demonstrated that the increased concentration of 8-oxo-dGTP and 8-oxo-GTP in MTH1 -knockdown cells and exogenous 8-oxoGTP, rather than 8-oxo-dGTP, could significantly increase the phosphorylation of STAT3. In conclusion, this work indicates that MTH1 inhibition increased the proportion of breast cancer stem cells (BCSCs) and promoted stemness properties in MCF7 cells. [Display omitted] • MTH1 inhibition shows a decreased proliferative ability due to oxidized dNTP. • MTH1 inhibition promotes stemness properties through upregulation of STAT3. • Exogenous 8-oxoGTP significantly increases the phosphorylation of STAT3. [ABSTRACT FROM AUTHOR]

Background The previous studies investigated the scaffold as mechanical carrier to stem cells or growth factors as transplantation therapy. This study aimed to investigate the effect of nanofiber scaffold on immunomodulatory properties of MSCs in osteoarthritis (OA) and the mechanism of this effect. Methods Collagen coated PCL nanofiber was prepared and examined by SEM. Mononuclear cells (MNCs) from the peripheral blood (P.B) and synovial fluid (SF) cultured with BMMSCs after stimulation by PHA using two types of co-culture (MSCs/Nano- and MSCs/nano+). Using flowcytometry, CD4, CD8, CD28, activation markers CD38 and CD23, regulatory CD4CD25high foxp3, CD14, IL17 by ELISA and cell proliferation using CCK-8 were assessed. ICAM mRNA was assessed by qRT-PCR and flowcytometry. Results MSCs/nano+ showed more significant inhibitory effect of MSCs regarding CD4, CD8, CD28, CD28 MFI, CD38, CD38 MFI, CD23, CD23 MFI, CD14, IL17 (p < 0.001) and higher level of Treg cells (p < 0.001) than nano− with difference regarding cell proliferation (p = 0.275). Also, the immunomodulatory effect of MSCs culture on nano+ on lymphocyte separated from S·F of osteoarthritis patients more powerful than nano−. The secretory function of MSCs culture on nano+ is more beneficial than that of nano-. Nano+ showed higher level of ICAM mRNA and CD54 than nano−. Conclusion The present study proved a novel function of nanofiber by increasing the immunomodulatory effect of the MSCs in osteoarthritis through increased the ICAM expression and this promising for increase the success of MSCs transplantation in OA. [ABSTRACT FROM AUTHOR]

• CRP levels frequently remain within the normal range despite active psoriatic disease. • Biomarker levels associated with moderate/high disease activity included SAA, IL-8, IP10, M-CSF, SCGF-β, and SDF-1α. • Use of multi-biomarker panel for assessing disease activity may provide better discrimination of disease activity. To investigate the correlation of serum protein biomarkers and disease activity in patients with PsA. 176 patients fulfilled the CASPAR (ClASsification criteria for Psoriatic ARthritis) were recruited in this cross-sectional study. The level of 48 protein biomarkers, cartilage and bone turn-over markers were assessed. The patients were randomly divided into a derivation-cohort and a validation-cohort at a ratio of 7:3. Patients were further categorized based on their disease activity states using cDAPSA (remission/low disease activity and moderate/high disease activity). Least absolute shrinkage and selection operator (LASSO) was used to select biomarkers which were associated with moderate/high disease activity in the derivation cohort. Receiver operating characteristic (ROC) curve, GiViTI calibration belt were used to assess the performance of the model in both cohorts. The cohort [age: 55.5 (44.0–62.75) years, male: 80 (45.5 %)] had moderate disease activity [DAPSA: 15.9 (8.3–26.9); PASI: 3.2 (0.5–6.8)]. 101 PsA patients (57.4 %) had clinical DAPSA moderate/high disease activity. Biomarker levels associated with moderate/high disease activity included SAA (Serum amyloid A), IL-8 (Interleukin 8), IP10 (Interferon gamma-induced protein 10)/CXCL10, M-CSF (Macrophage colony-stimulating factor), SCGF-β (Stem cell growth factor), SDF-1α (Stromal cell-derived factor 1α)/CXCL12. The model's equation including the 6 biomarker levels was applied to the validation-cohort. The area under the ROC curve (AUC) for discriminating moderate/high disease activity was 0.802 and 0.835 for the derivation-and-validation-cohorts, respectively. The multi-biomarkers panel model had higher-AUC when compared with that of C-reactive protein (CRP) (AUC = 0.727, p = 0.022). The P-values of calibration charts in the two sets were 0.902 and 0.123. The multi-biomarkers panel demonstrated the ability to discriminate patients with moderate/high disease activity from those with low disease activity/remission. [ABSTRACT FROM AUTHOR]

[Display omitted] • Normal wound healing progresses through homeostasis, inflammatory proliferative and remodeling phases. • Impaired wound healing in DFU is characterized by Hyperglycemia, Hypoxia, Bacterial infection, prolonged Inflammation and impaired Angiogenesis. • Collagen III is synthesized in the early stages of wound healing and is replaced by collagen I. • Collagen resolves inflammation, promote angiogenesis and tissue remodeling. • Collagen can be extracted from different sources and formulated into films, sponges, hydrogels, powders and nanofibers. Diabetic foot ulcer (DFU), a chronic wound with complex pathophysiology and immune cell imbalance is the major health concern as it takes longer healing time compared to normal wound healing. Collagen wound dressings have important role in DFU healing as these dressings improve angiogenesis, reduces inflammation time and aids in the regeneration of damaged tissue, maintains a moist wound environment by promoting optimal conditions for cell activity. These dressings are formulated with bovine, avian, porcine, marine, human based or recombinant sources such as plant based, insect based or microorganisms. Animal based biomaterials have concern of immunogenicity and purification while marine based and recombinant sources are better alternatives for these issues. Chemical modifications with other bioactive materials, reduces immunogenicity and the healing time, increase antioxidant activity, modulate inflammatory responses, improve wound microenvironment and promote broad spectrum antimicrobial property. Additionally, incorporation of mesenchymal stem cells and growth factors can generate various cells essential to wound healing, such as dermal fibroblasts (DFs), endothelial cells, and keratinocytes. This review summarizes the role of collagen wound dressing in DFU wound healing, to provide the conceptual understanding of the research and development of collagen as a potential biomaterial for diabetic wound healing. [ABSTRACT FROM AUTHOR]

[Display omitted] The hypoxic microenvironment in esophageal carcinoma is an important factor promoting the rapid progression of malignant tumor. This study was to investigate the lactylation of Axin1 on glycolysis in esophageal carcinoma cells under hypoxia exposure. Hypoxia treatment increases pan lysine lactylation (pan-kla) levels of both TE1 and EC109 cells. Meanwhile, ECAR, glucose consumption and lactate production were also upregulated in both TE1 and EC109 cells. The expression of embryonic stem cell transcription factors NANOG and SOX2 were enhanced in the hypoxia-treated cells. Axin1 overexpression partly reverses the induction effects of hypoxia treatment in TE1 and EC109 cells. Moreover, lactylation of Axin1 protein at K147 induced by hypoxia treatment promotes ubiquitination modification of Axin1 protein to promote glycolysis and cell stemness of TE1 and EC109 cells. Mutant Axin1 can inhibit ECAR, glucose uptake, lactate secretion, and cell stemness in TE1 and EC109 cells under normal or hypoxia conditions. Meanwhile, mutant Axin1 further enhanced the effects of 2-DG on inhibiting glycolysis and cell stemness. Overexpression of Axin1 also inhibited tumor growth in vivo , and was related to suppressing glycolysis. In conclusion, hypoxia treatment promoted the glycolysis and cell stemness of esophageal carcinoma cells, and increased the lactylation of Axin1 protein. Overexpression of Axin1 functioned as a glycolysis inhibitor, and suppressed the effects of hypoxia exposure in vitro and inhibited tumor growth in vivo. Mechanically, hypoxia induces the lactylation of Axin1 protein and promotes the ubiquitination of Axin1 to degrade the protein, thereby exercising its anti-glycolytic function. [ABSTRACT FROM AUTHOR]

Peripheral blood stem cells (PBSC) mobilization with granulocyte colony stimulating factor (G-CSF) for healthy donors is generally performed at 5th day. However, earlier collection is sometimes feasible, raising the question of whether to initiate apheresis early to limit further G-CSF exposure, while considering the risk of mobilization failure. In the current study, we examined the factors predicting successful 4th day collection and developed a model that can be used practically. The study was carried out by obtaining the data of PBSC mobilizations performed between January 2009 and September 2022 in our transplantation center. A total of 141 healthy donors with a median donor age of 32 (18–64) were included. Adequate mobilization was achieved in 115 (81.6 %) patients. Median peripheral CD34 + cell count was 69.4/μL in the adequate mobilization group and 46/μL in the mobilization failure group (p < 0001). Multivariate analysis revealed that donor/recipient weight ratio and the 4th day peripheral CD34 + cell count≥ 50/μL were independent markers for 4th day collection success. A predictive model of our center including these parameters was available with 0.765 sensitivity and 0.968 specificity [(AUC):0.948 (95 % CI, 0.90–0.99), p < 0.001]. The result of the current study shows that peripheral 4th day collection can be performed in selected donors, taking into account peripheral CD34+ cell count and donor/recipient weight ratio. In addition, using these indicators, new predictive models can be created that may assist clinicians in daily practice. [ABSTRACT FROM AUTHOR]

To detect human neutrophil (n), monocyte (m), erythroblast (E), megakaryocyte (Mk) colonies is difficult, mostly because in vitro culture conditions for human megakaryocyte progenitors are poorly defined. We developed a new serum-free culture with thrombopoietin receptor agonist lusutrombopag (LPAG). CD34+CD110lowCD117low cells were isolated from umbilical cord blood and cultured, followed by flow cytometry analysis to detect CD14-CD16+ n, CD14+CD16- m, CD235a+ E, and CD41+ Mk. The number of Mk in culture with recombinant human serum albumin was significantly greater than that with polyvinyl alcohol. The number of Mk in culture with LPAG was significantly greater than that with thrombopoietin. There was no synergistic effect between stem cell factor (SCF) and LPAG. LPAG alone supported Mk differentiation from phenotypically defined common myeloid progenitors (CMP), megakaryocyte/erythrocyte progenitors (MEP), and Mk progenitors, but not hematopoietic stem cells (HSC). Since LPAG also supported nmE differentiation in the presence of SCF and interleukin-3 (IL-3), the minimum combination of SCF, LPAG, IL-3, and erythropoietin supported nmEMk differentiation. Single-cell colony assay showed that nmEMk colony-forming cells were detectable in 5-10 % of phenotypically defined CMP and MEP, but not HSCs, suggesting the presence of functional CMP in these populations. Whether CMP are present is controversial. Our culture conditions are useful for further characterization of human CMP. [ABSTRACT FROM AUTHOR]

Hematopoietic stem cell (HSC) is a clinically relevant blood-generating stem cell where HSC transplantation (HSCT) remains only a curative option for multiple blood-related disorders. HSC applications are, however, hampered partially because HSC cannot expand stably ex vivo. Stable ex vivo HSC expansion would address current challenges including an engraftment failure in umbilical cord blood (UCB)-HSCT and limited success of genetic modifications in HSCs. Our group previously developed a stable ex vivo mouse HSC expansion protocol using a polymer-based medium with thrombopoietin (TPO) and stem cell factor (SCF) under hypoxic conditions. Extending this approach to human HSCs, an cytokine-free culture protocol comprising 740 Y-P, a PI3K activator as an SCF replacement, and TPO agonist along with UM171 have been reported. However, the expansion of human HSCs was not so robust as that of mouse HSCs. Here, we applied findings from the stable mouse HSC expansion to improve human HSC expansion. We found that a culture under 5% O2 improves human CD34+ UCB expansion. A screening that targets TPO/SCF signaling with >100 small molecules/peptides identified that adding SCF with 740 Y-P enhances CD34+ UCB expansion (n=7 donors; mean fold change: 105.2, ranging from 36.2 - 402.3, in total viable cells over a 28-day culture). We confirmed the expansion of human HSCs in a xenograft model (mean human chimerism of 22.9% [14-day expanded] vs 0.96% [unmanipulated] 16 weeks post-transplantation). HDR-based targetted knockin of a genetic cassset that reports endogenous expression of HLF, a HSC-specific transcription factor, revealed the enrichment of HLF-positive cells following the culture, indicating the successful gene modification targeting HSCs. In summary, our study presents a promising advancement in HSC applications through hypoxic SCF-supplemented human HSC expansion. [ABSTRACT FROM AUTHOR]

Within the mammalian hematopoietic system, Granulocyte Colony-Stimulating Factor (G-CSF) is one of the master regulators of the myeloid lineage, controlling proliferation, maturation, and function of granulocyte progenitor, precursor, and mature cells. Currently, G-CSF is widely used in clinical settings to stimulate granulopoiesis in neutropenic patients, and to promote hematopoietic stem/progenitor cell mobilization in hematopoietic transplant donors. Interestingly, recent reports using murine models indicate that high levels of G-CSF affect the erythroid lineage, inducing an anemic phenotype in vivo. The processes and mechanisms underlying such observations are not fully understood. Thus, in trying to gain insight into this issue, in the present study, we have analyzed the effects of G-CSF on the in vitro biology of very early erythroid progenitors (megakaryocytic-erythroid progenitors [MEPs]; CD34+ CD38+ CD45RA- CD135- CD10- cells). MEPs were purified (using FACS) from human cord blood or mobilized peripheral blood and cultured in serum-free liquid cultures in the presence of Stem Cell Factor (SCF) and G-CSF (without/with erythropoietin [EPO]) for 12 days. Our results show that G-CSF was able to stimulate MEPs proliferation, although at a much lower level compared with EPO. Interestingly, G-CSF significantly altered MEPs differentiation, inhibiting the generation of erythroid progenitor and precursor cells, and promoting a switch towards the myeloid lineage. It induced de novo generation of myeloid progenitors (CFU-G and CFU-M), as well as myeloid precursor/mature cells (mainly granulocytic CD15+ CD66b+ cells). Our study indicates that regulation of primitive erythroid progenitors' differentiation may be more complex than previously thought. The in vivo physiological relevance of these findings should be further explored. [ABSTRACT FROM AUTHOR]

Bi-allelic (bi) mutations in the myeloid transcription factor CEBPA are found in about 5% of acute myeloid leukemia (AML) patients and frequently co-occur with mutations in TET2, WT1 and GATA2. Co-occurrence with TET2 and WT1 mutations give a worse overall survival whereas co-occurrence with GATA2 mutations improve overall survival. To study the underlying molecular mechanisms, we modeled these mutations in primary human bone marrow (BM) hematopoietic stem and progenitor cells (HSPCs) using CRISPR/Cas9 gene editing. Targeting the N-terminal part of CEBPA using Cas9 ribonucleoproteins resulted in loss of the full-length CEBPA protein (p42) and marked upregulation of the shorter p30 isoform as seen in patients. CEBPAbi mutations resulted in loss of monocytic colonies and increased serial replating capacity in CFU assays. CEBPAbi combined with TET2 or WT1 loss-of-function mutations resulted in leukemic transformation and uncontrolled cell proliferation in a mouse stromal-5 co-culture assay. Transplantation of CEBPAbi-mutated HSPCs into immunodeficient mice expressing human cytokines (NSGS mice) revealed a myeloid skewing, and when combined with TET2 or WT1 mutations, leukemic transformation. Interestingly, while CEBPAbi- and co-mutated HSPCs could engraft in the BM of NSG mice, we did not observe dissemination to the peripheral blood and no leukemic transformation. Subsequent cytokine dropout experiments revealed a dependency on human stem cell factor in CEBPAbi leukemia development. Proteome analysis showed that protein differences were mainly driven by CEBPAbi mutations and strengthened by loss of TET2 or WT1. These data suggest that an altered chromatin state due to TET2 or WT1 loss further enhances CEBPA p30-mediated transcription. We are currently performing scRNAseq and ATACseq to further substantiate these findings and decipher the underlying mechanisms. [ABSTRACT FROM AUTHOR]

Immunoglobulin A nephropathy (IgAN) is an autoimmune disease characterized by the production of galactose‑deficient IgA1 (Gd‑IgA1) and the deposition of immune complexes in the kidney. Exploring the landscape of immune dysregulation in IgAN is valuable for pathogenesis and disease treatment. We conducted Mendelian randomization (MR) to assess the causal correlations between inflammation and IgAN. Based on available genetic datasets, we investigated potential causal links between inflammation and the risk of IgAN using two-sample MR. We used genome-wide association study (GWAS) summary statistics of 5 typical inflammation markers, 41 inflammatory cytokines, and 731 immune cell signatures, accessed from the public GWAS Catalog. The primary method employed for MR analysis was Inverse Variance Weighted (IVW). To confirm consistency across results, four supplementary MR methods were also conducted: MR-Egger, Weighted Median, Weighted Mode, and Simple Mode. To assess pleiotropy, we used the MR-Egger regression intercept test and Mendelian Randomization Pleiotropy RESidual Sum and Outlier (MR-PRESSO) test. Cochrane's Q statistic was applied to evaluate heterogeneity. Additionally, the stability of the MR findings was verified through the leave-one-out sensitivity analysis. This study revealed that interleukin-7 (IL-7) and stem cell growth factor beta (SCGF-β) were possibly associated with the risk of IgAN according to the IVW approach, with estimated odds ratios (OR) of 1.059 (95 % confidence interval [CI] 1.015 to 1.104, P = 0.008) and 1.043 (95 % CI 1.002 to 1.085, P = 0.037). Five immune traits were identified that might be linked to IgAN risk, each with P -values below 0.01, including natural killer T %T cell (OR = 1.058, 95 % CI: 1.020 to 1.097, P = 0.002), natural killer T %lymphocyte (OR = 1.055, 95 % CI: 1.016 to 1.096, P = 0.006), CD25++ CD8+ T cell %T cell (OR = 1.057, 95 % CI: 1.016 to 1.099, P = 0.006), CD3 on effector memory CD4+ T cell (OR = 1.045, 95 % CI: 1.019 to 1.071, P = 0.001), and CD3 on CD28+ CD45RA+ CD8+ T cell (OR = 1.042, 95 % CI: 1.016 to 1.068, P = 0.001). CD4 on central memory CD4+ T cell might be a protective factor for IgAN (OR = 0.922, 95 % CI: 0.875 to 0.971, P = 0.002). Moreover, IgAN may be implicated in a high risk of elevated granulocyte colony-stimulating factor (G-CSF) (OR = 1.114, 95 % CI 1.002 to 1.239, P = 0.046). Our study revealed exposures among typical inflammation markers, inflammatory cytokines, and immune cell signatures that may potentially linked to IgAN risk by MR analysis. This insight may advance our understanding of the etiology of IgAN and support the development of targeted therapeutic strategies. [ABSTRACT FROM AUTHOR]

Periodontitis is the primary cause of tooth loss, but there is no effective treatment to repair inflammatory bone loss in periodontitis. Exosomes emerge as essential paracrine factors of mesenchymal stem cells (MSCs) that mediated tissue regeneration. Here, we investigated the potential of exosomes secreted by periodontal ligament stem cells (PDLSCs) as therapeutics for the bone defect in periodontitis. Exosomes secreted from PDLSCs derived from healthy periodontal ligaments (h-PDLSCs) and their function were evaluated on PDLSCs isolated from the inflammatory periodontal ligament of periodontitis patients (i-PDLSCs). Treatment of exosomes of h-PDLSCs led to an increase in the formation of mineralized nodules and the expressions of osteogenic genes and proteins in i-PDLSCs. Mechanistically, h-PDLSCs-exosomes suppressed the over-activation of canonical Wnt signaling to recover the osteogenic differentiation capacity of i-PDLSCs. To evaluate the therapeutic of exosomes on inflammatory bone loss, h-PDLSCs-exosomes loaded with Matrigel or β-TCP were employed to repair bone defects in rat models of periodontitis. Compared to the vehicle-treated control group, h-PDLSCs-exosomes-treated rats resulted in more bone formation in the defect of alveolar bone. In conclusion, these results demonstrated that exosomes derived from healthy PDLSCs could rescue the osteogenesis capacity of endogenous stem cells under an inflammatory environment and promote regeneration of alveolar bone. Our findings suggest that MSCs-derived exosome is an effective and practical cell-free MSC therapeutic for the treatment of periodontitis. There is no effective treatment to repair inflammatory bone loss in periodontitis. As essential paracrine factors of PDLSCs, exosomes might mediate tissue regeneration during stem cell therapy. Here, we reported that exosomes secreted from healthy PDLSCs promoted the osteogenic differentiation of PDLSCs derived from periodontitis tissue. Healthy PDLSCs-exosomes treatment resulted in accelerated bone formation in the defect of alveolar bone in rat models of periodontitis. Mechanistically, h-PDLSCs-exosomes suppressed the over-activation of canonical Wnt signaling to recover the osteogenic differentiation capacity of inflammatory PDLSCs. These findings suggest that MSCs-derived exosome is an effective and practical cell-free MSC therapeutic for the treatment of periodontitis. [Display omitted] [ABSTRACT FROM AUTHOR]

Adult organs are vascularized by specialized blood vessels. In addition to inter-organ vascular heterogeneity, each organ is arborized by structurally and functionally diversified populations of endothelial cells (ECs). The molecular pathways that are induced to orchestrate inter- and intra- organ vascular heterogeneity and zonation are shaped during development and fully specified postnatally. Notably, intra-organ specialization of ECs is associated with induction of angiocrine factors that guide cross-talk between ECs and parenchymal cells, establishing co-zonated vascular regions within each organ. In this review, we describe how microenvironmental tissue-specific biophysical, biochemical, immune, and inflammatory cues dictate the specialization of ECs with zonated functions. We delineate how physiological and biophysical stressors in the developing liver, lung, and kidney vasculature induce specialization of capillary beds. Deciphering mechanisms by which vascular microvasculature diversity is attained could set the stage for treating regenerative disorders and promote healing of organs without provoking fibrosis. ECs secrete angiocrine factors to regulate stem cell self-renewal, differentiation and maturation. Organ specific extra-vascular cues mediate EC specialization mediated by TF activation. ETV2 - RVECs vascularize organoids and organo-typically adapt to the microenvironment. [ABSTRACT FROM AUTHOR]

[Display omitted] Fate determination of germline stem cells remains poorly understood at the chromatin structure level. Our research hopes to develop successful offspring production of ovarian organoids derived from spermatogonial stem cells (SSCs) by defined factors. The offspring production from oocytes transdifferentiated from mouse SSCs with tracking of transplanted SSCs in vivo, single cell whole exome sequencing, and in 3D cell culture reconstitution of the process of oogenesis derived from SSCs. The defined factors were screened with ovarian organoids. We uncovered extensive chromatin reorganization during SSC conversion into induced germline stem cells (iGSCs) using high throughput chromosome conformation. We demonstrate successful production of offspring from oocytes transdifferentiated from mouse spermatogonial stem cells (SSCs). Furthermore, we demonstrate direct induction of germline stem cells (iGSCs) differentiated into functional oocytes by transduction of H19 , Stella , and Zfp57 and inactivation of Plzf in SSCs after screening with ovarian organoids. We uncovered extensive chromatin reorganization during SSC conversion into iGSCs, which was highly similar to female germline stem cells. We observed that although topologically associating domains were stable during SSC conversion, chromatin interactions changed in a striking manner, altering 35% of inactive and active chromosomal compartments throughout the genome. We demonstrate successful offspring production of ovarian organoids derived from SSCs by defined factors with chromatin reorganization. These findings have important implications in various areas including mammalian gametogenesis, genetic and epigenetic reprogramming, biotechnology, and medicine. [ABSTRACT FROM AUTHOR]

After aortic coarctation (CoA) repair, patients still suffer from cardiovascular complications. The aim of this study was to measure cardiovascular markers, intima-media thickness (IMT) and plaques in controls and patients with CoA. Sixty-four patients with CoA (66% male, mean age 48 ± 15 years) and controls (54% men, mean age 47 ± 16 years) underwent ultrasound of their arteries. A multiplex platform to analyze circulating blood levels biomarkers reflecting inflammation, tissue remodeling and repair was used. In men following CoA repair, a significantly increased carotid bulb IMT was observed in comparison to the control group (1.05 [0.72–1.24] vs. 0.67 [0.59–0.95] mm; p = 0.003). Median common carotid artery (CCA) IMT was increased in men compared to controls (0.82 [0.61–0.97] mm vs. 0.58 [0.53–0.76] mm, p < 0.003) and in women compared to controls (0.83 [0.70–0.92] vs. 0.60 [0.55–0.69], p < 0.004). CoA demonstrated an independent association with IMT in both men and women. Men with CoA were also more likely to have a plaque in their carotid arteries (p = 0.010). In women with CoA, we observed significantly lower levels of stem cell factor (SCF, p = 0.004) while in men with CoA we observed significantly lower levels of matrix metalloproteinase-3 (MMP-3, p = 0.048), tumor necrosis factor receptor 1 (TNF-R1, p = 0.032), tumor necrosis factor receptor superfamily member 10B (TRAIL-R2, p = 0.019) and monocyte chemotactic protein 1 (MCP-1, p = 0.015). This is the first study to show that despite successful CoA repair, patients have more carotid atherosclerosis than can be explained by changes in tissue remodeling and repair. [Display omitted] • Generalized vascular disease in patients with aortic coarctation. • A relationship between atherosclerosis and markers for vascular repair and apoptosis. • Apoptosis and repair mechanisms involved in cardiovascular risk. [ABSTRACT FROM AUTHOR]

There is increasing evidence that long noncoding RNAs (lncRNAs) are among the main regulatory factors of stem cell maintenance and differentiation. They act through various mechanisms and interactions with proteins, DNA, and RNA. This heterogeneity in function increases the capabilities of the lncRNome toolkit but also makes it difficult to predict the function of novel lncRNAs or even rely on biological information produced in animal models. As lncRNAs are species- and tissue-specific, the recent technical advances in self-renewal and differentiation of human embryonic stem cells (ESCs) make these cells the ideal system to identify key regulatory lncRNAs and study their molecular functions. Here we provide an overview of the functional versatility of lncRNA mechanistic heterogeneity in regulating pluripotency maintenance and human differentiation. Embryonic differentiation at the gastrulation stage is a very tightly regulated process in which many long noncoding RNAs (lncRNAs) have been demonstrated to play a crucial role in gene expression modulation. lncRNAs can regulate cellular functions at the transcriptional, post-transcriptional, translational, and post-translational levels. lncRNAs exert a large repertoire of regulatory functions both in the nucleus and in the cytoplasm. [ABSTRACT FROM AUTHOR]

Previous views of reactive oxygen species (ROS) depicted them as harmful byproducts of metabolism as uncontrolled levels of ROS can lead to DNA damage and cell death. However, recent studies have shed light into the key role of ROS in the self-renewal or differentiation of the stem cell. The interplay between ROS levels, metabolism, and the downstream redox signaling pathways influence stem cell fate. In this review we will define ROS, explain how they are generated, and how ROS signaling can influence transcription factors, first and foremost forkhead box-O transcription factors, that shape not only the cellular redox state, but also stem cell fate. Now that studies have illustrated the importance of redox homeostasis and the role of redox signaling, understanding the mechanisms behind this interplay will further shed light into stem cell biology. [Display omitted] • Stem cells exhibit specialized energy metabolism that maintains a healthy redox state. • Oxidative stress aligns with stemness loss and initiation of differentiation. • Stem cells feature specialized redox sensors that conserve stemness. • FOXO transcription factors serve as stem cell redox sensors. • The outcome of FOXO activation might differ between ASCs and PSCs. [ABSTRACT FROM AUTHOR]

Introduction: Treatment of infected non-unions of the tibia is a challenging problem. The cornerstones of optimal infected non-union treatment consist of extensive debridement, fracture fixation, antimicrobial therapy and creation of an optimal local biological bone healing environment. The combination of S53P4 bioactive glass (BAG), as osteostimulative antibacterial bone graft substitute, and bone marrow aspirate concentrate (BMAC) for the implantation of mesenchymal stem cells and growth factors might be a promising combination. In this paper, preliminary results of a new treatment algorithm for infected non-unions of the tibia is presented.Methods: In this retrospective case series patients with infected non-unions of the tibia are treated according to a new treatment algorithm. Patients are treated with extensive debridement surgery, replacement of the osteosynthesis and implantation of S53P4 BAG and BMAC in a one-stage or two-stage procedure based on non-union severity. Subsequently patients are treated with culture based antibiotic therapy and followed until union and infection eradication.Results: Five patients with an infected non-union were treated, mean age was 55, average NUSS-score was 44 and the average segmental bone defect was 4.6cm. One patient was treated in a one-stage procedure and four patients in a two-stage induced membrane-, or "Masquelet"-procedure. On average, 23 ml S53P4 BAG and 6.2 ml BMAC was implanted. The mean follow-up period was 13.6 months and at the end of follow-up all patients had clinical consolidation with an average RUST-score of 7.8 and complete eradication of infection.Discussion: These early data on the combined implantation of S53P4 BAG and BMAC in treatment of infected non-unions shows promising results. These fracture healing results and eradication rates resulted in promising functional recovery of the patients. To substantiate these results, larger and higher quality studies should be performed. [ABSTRACT FROM AUTHOR]

NOTCH2 is expressed in pituitary stem cells and is necessary for stem cell maintenance, proliferation, and differentiation. However, the pathways NOTCH2 engages to affect pituitary development remain unclear. In this study, we hypothesized that glycoprotein hormone subunit A2 (GPHA2), a corneal stem cell factor and ligand for the thyroid stimulating hormone receptor (TSHR), is downstream of NOTCH2 signaling. We found Gpha2 is expressed in quiescent pituitary stem cells by RNAscope in situ hybridization and scRNA seq. In Notch2 conditional knockout pituitaries, Gpha2 mRNA is reduced compared with control littermates. We then investigated the possible functions of GPHA2. Pituitaries treated with a GPHA2 peptide do not have a change in proliferation. However, in dissociated adult pituitary cells, GPHA2 increased pCREB expression and this induction was reversed by co-treatment with a TSHR inhibitor. These data suggest GPHA2 is a NOTCH2 related stem cell factor that activates TSHR signaling, potentially impacting pituitary development. • GPHA2 is expressed in quiescent pituitary stem cells. • Gpha2 expression is suppressed without Notch2 signaling. • GPHA2 causes CREB phosphorylation in pituitary cultures. • GPHA2 can signal through TSH receptor. [ABSTRACT FROM AUTHOR]

The developmental origin of blood-forming hematopoietic stem cells (HSCs) is a longstanding question. Here, our non-invasive genetic lineage tracing in mouse embryos pinpoints that artery endothelial cells generate HSCs. Arteries are transiently competent to generate HSCs for 2.5 days (∼E8.5–E11) but subsequently cease, delimiting a narrow time frame for HSC formation in vivo. Guided by the arterial origins of blood, we efficiently and rapidly differentiate human pluripotent stem cells (hPSCs) into posterior primitive streak, lateral mesoderm, artery endothelium, hemogenic endothelium, and >90% pure hematopoietic progenitors within 10 days. hPSC-derived hematopoietic progenitors generate T, B, NK, erythroid, and myeloid cells in vitro and, critically, express hallmark HSC transcription factors HLF and HOXA5-HOXA10 , which were previously challenging to upregulate. We differentiated hPSCs into highly enriched HLF + HOXA + hematopoietic progenitors with near-stoichiometric efficiency by blocking formation of unwanted lineages at each differentiation step. hPSC-derived HLF + HOXA + hematopoietic progenitors could avail both basic research and cellular therapies. [Display omitted] • Lineage tracing demonstrates that artery endothelial cells (ECs) form HSCs in vivo • Stepwise hPSC differentiation into artery ECs and then hematopoietic progenitors • Rapid and efficient generation of >90% pure hematopoietic progenitors in 10 days • hPSC-derived hematopoietic progenitors express core HSC markers HLF and HOXA5-10 Fowler and Zheng et al. show that artery endothelial cells give rise to hematopoietic stem cells in the mouse embryo. Leveraging this knowledge, they differentiate human pluripotent stem cells into artery endothelial cells and, subsequently, hematopoietic progenitors that express HLF and HOXA5-10 , which are hallmark hematopoietic stem cell transcription factors. [ABSTRACT FROM AUTHOR]

The immune microenvironment constructed by tumor-infiltrating immune cells and the molecular phenotype defined by hormone receptors (HRs) have been implicated as decisive factors in the regulation of breast cancer (BC) progression. Here, we found that the infiltration of mast cells (MCs) informed impaired prognoses in HR(+) BC but predicted improved prognoses in HR(−) BC. However, molecular features of MCs in different BC remain unclear. We next discovered that HR(−) BC cells were prone to apoptosis under the stimulation of MCs, whereas HR(+) BC cells exerted anti-apoptotic effects. Mechanistically, in HR(+) BC, the KIT ligand (KITLG), a major mast cell growth factor in recruiting and activating MCs, could be transcriptionally upregulated by the progesterone receptor (PGR), and elevate the production of MC-derived granulin (GRN). GRN attenuates TNFα-induced apoptosis in BC cells by competitively binding to TNFR1. Furthermore, disruption of PGR-KITLG signaling by knocking down PGR or using the specific KITLG-cKIT inhibitor iSCK03 potently enhanced the sensitivity of HR(+) BC cells to MC-induced apoptosis and exerted anti-tumor activity. Collectively, these results demonstrate that PGR-KITLG signaling in BC cells preferentially induces GRN expression in MCs to exert anti-apoptotic effects, with potential value in developing precision medicine approaches for diagnosis and treatment. • The infiltration of mast cells impaired prognoses in (HR) (+) BC but predicted improved prognoses in HR(−) BC. • HR(−) BC cells were prone to apoptosis under the stimulation of MCs, whereas HR(+) BC cells exerted anti-apoptotic effects. • PGR transcriptionally up regulates the expression of KITLG in HR(+) BC cells. • KITLG elevates the infiltration of MCs and stimulates GRN expression in MCs. • MC-derived GRN can competitively bind to TNFR1, thus inhibiting the TNF-α apoptotic pathway. [ABSTRACT FROM AUTHOR]

Retinal ischemia is a significant factor in various vision-threatening diseases, but effective treatments are currently lacking. This study explores the potential of stem cell factor (SCF) in regulating the neurovascular unit as a therapeutic intervention for retinal ischemic diseases. A chronic retinal ischemia model was established in Brown Norway rats using bilateral common carotid artery occlusion (BCCAO). Subsequent SCF treatment resulted in a remarkable recovery of retinal function, as indicated by electroretinogram, light/dark transition test, and optokinetic head tracking test results. Histological examination demonstrated a significant increase in the number of retinal neurons and an overall thickening of the retina. Immunofluorescence confirmed these findings and further demonstrated that SCF treatment regulated retinal remodeling. Notably, SCF treatment ameliorated the disrupted expression of synaptic markers in the control group's BCCAO rats and suppressed the activation of Müller cells and microglia. Retinal whole-mount analysis revealed a significant improvement in the abnormalities in retinal vasculature following SCF treatment. Transcriptome sequencing analysis revealed that SCF-induced transcriptome changes were closely linked to the Wnt7 pathway. Key members of the Wnt7 pathway, exhibited significant upregulation following SCF treatment. These results underscore the protective role of SCF in the neurovascular unit of retinal ischemia rats by modulating the Wnt7 pathway. SCF administration emerges as a promising therapeutic strategy for retinal ischemia-related diseases, offering potential avenues for future clinical interventions. • SCF effectively restored retinal function in chronic ischemia. • SCF modulated the neurovascular unit on neurons, glial cells, and vascular cells. • The Wnt7 pathway played a crucial role in providing protection. [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease (IBD) is a major subtype of chronic enteropathies in dogs and cats. Conventional drugs such as immunomodulatory medicines as glucocorticoids and/or other anti-inflammatory are mainly applied for treatment. However, these drugs are not always effective to maintain remission from IBD and are limited by unacceptable side effects. Hence, more effective and safe therapeutic options need to be developed. Mesenchymal stem cells (MSCs) are multipotent stem cells with a self-renewal capacity, and have immunomodulatory, anti-inflammatory, anti-fibrotic, and tissue repair properties. Therefore, the application of MSCs as an alternative therapy for IBD has great potential in veterinary medicine. The efficacy of adipose tissue-derived MSC (ADSC) therapy for IBD in dogs and cats has been reported, including numerous studies in animal models. However, treatment outcomes in clinical trials of human IBD patients have not been consistent with preclinical studies. MSC-based therapy for various diseases has received widespread attention, but various problems in such therapy remain, among which no consensus has been reached on the preparation and treatment procedures for MSCs, and cellular heterogeneity of MSCs may be an issue. This review describes the current status of ADSC therapy for canine and feline IBD and summarizes the cellular heterogeneity of canine ADSCs, to highlight the necessity for further reduction or elimination of MSCs heterogeneity and standardization of MSC-based therapies. • Mesenchymal stem cell (MSC) therapy is expected for inflammatory bowel disease. • Heterogeneity of MSCs is a cause of unstable therapeutic effects. • Overcoming the heterogeneity of MSCs is challenged in veterinary medicine. [ABSTRACT FROM AUTHOR]

• β-sitosterol is the active component of Amomum villosum to promote defecation. • β-sitosterol exerts laxative effects by activating the ADRA1A-MLC signalling pathway. • β-sitosterol relieves constipation by increasing gastrointestinal motility-related factors. Constipation is a prevalent gastrointestinal disorder, with its prevalence showing an annual upward trend. There are many factors involved in the occurrence of constipation, such as abnormal smooth muscle contraction and disorders of gastrointestinal hormone secretion. Amomum villosum (A. villosum) has been proven to be effective in improving digestive system diseases, but there is no report on improving constipation. Therefore, we used network pharmacology prediction combined with animal experiments to explore the key active components of A. villosum and their pharmacological mechanisms. The results of network pharmacological prediction showed that β-sitosterol was the key laxative compound of A. villosum , which may play a laxative role by activating the adrenoceptor alpha 1 A-myosin light chain (ADRA1A-MLC) pathway. Further animal experiments showed that β-sitosterol could significantly shorten the time to first black stool; increase faecal weight, faecal number, and faecal water content; and promote gastrointestinal motility. β-sitosterol may promote intestinal motility by upregulating the expression of ADRA1A and myosin light chain 9 (Myl9) mRNA and protein in the colon, thereby activating the ADRA1A-MLC signalling pathway. In addition, it is possible to improve constipation symptoms by regulating serum neurotransmitters and gastrointestinal motility-related factors, such as the serum content of 5-hydroxytryptamine (5-HT) and acetylcholinesterase (AchE) and the mRNA expression of 5-hydroxytryptamine receptor 4 (5-HT4) , stem cell factor (SCF), stem cell factor receptor (c-Kit) and smooth muscle myosin light chain kinase (smMLCK) in the colon. These results lay a foundation for the application of A. villosum and β-sitosterol in constipation. [ABSTRACT FROM AUTHOR]

Synbiotic dietary supplements, as an effective means of regulating the gut microbiota, may have a beneficial effect on constipation. This study evaluated the effects of synbiotic yogurt containing konjac mannan oligosaccharides (KMOS) and Bifidobacterium animalis ssp. lactis BB12 (BB12) on constipated Kunming mice (the model group). Following administration of yogurt containing 2.0% KMOS and BB12 (YBK2.0), black fecal weight and number and gastrointestinal transit rate increased by 97.5, 106.3, and 55.7%, respectively, compared with the model group. Serum levels of excitability neurotransmitters (motilin, substance P, and acetylcholine) in the YBK2.0 group were increased by 139.7, 120.4, and 91.8%, respectively, and serum levels of inhibitory neurotransmitters (vasoactive intestinal peptide, nitric oxide, and acetylcholine) were decreased. Moreover, synbiotic yogurt supplementation significantly downregulated the expression of vasoactive intestinal peptide receptor 1 (VIPR1) and upregulated the expression of serotonin receptor 4 (5-HT4) in the colon, and enhanced the expression of the stem cell factor (SCF)/c-Kit pathway. Additionally, YBK2.0 treatment significantly regulated the community composition and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of gut microbiota, which were positively correlated with physiological parameters of constipation. Thus, supplementation with synbiotic yogurt composed of KMOS and BB12 could facilitate fecal excretion by regulating related pathways and the gut microbiota. These findings demonstrated that the synbiotic yogurt can be considered a functional food for alleviating constipation. [ABSTRACT FROM AUTHOR]

Pancreatic ductal adenocarcinoma (PDAC) shows poor prognosis and high malignancy due to the presence of cancer-initiating cells (CICs) and characteristics of the tumor microenvironment (TME). Organoids are useful for studying PDAC, and establishing organoids is dependent on stem cell growth factors, including Wnt signaling. Herein, using a conventional organoid culture system, we demonstrated that CD44(+)CD24(+) and CD44(+)CD24(+)EpCAM(+) CICs were enriched >65% in a PDAC patient-derived organoid. CICs expressing CD44 formed lumen structures by gathering into circles. Additionally, organoid-derived CD44(-) cancer cells were capable of organoid re-formation and could be re-programed as CD44-expressing CICs in the organoid culture system. To mimic a TME absent artificial stem cell growth factors, a PDAC organoid with vascular niche was established. CICs in the PDAC tumor organoid were maintained by paracrine effects and direct interactions with endothelial cells. Interestingly, CD44(+) cells in PDAC tumor tissue were detected primarily in the vascular niche. Inhibiting both Wnt and Notch signaling in endothelial cells suppressed organoid formation and the maintenance of CD24(+)CD44(+) CICs. Collectively, our results suggest that PDAC patient-derived organoids maintain CICs by interacting with endothelial cells via Wnt and Notch pathways. [ABSTRACT FROM AUTHOR]

Tumor angiogenesis is a major characteristic of renal cell carcinoma (RCC). Herein, we report a novel mechanism of how lncRNA and androgen receptor (AR) drive the Hedgehog pathway to promote tumor angiogenesis in RCC. We found that the high expression of lncRNA HOTAIR in RCC is associated with poor prognosis. Moreover, HOTAIR and AR form a feedback loop to promote the expression of each other. Interestingly, we also found that in RCC, HOTAIR is associated with the Hedgehog pathway, especially GLI2, via bioinformatics analysis. Furthermore, HOTAIR promotes GLI2 expression in the presence of AR. Mechanistically, HOTAIR interacts with AR and they cooperatively bind to GLI2 promoter and increase its transcription activity. We further confirmed how HOTAIR-AR axis regulates GLI2 expression by analyzing its function in RCC cells and found that HOTAIR and AR synergistically enhanced the expression of GLI2 downstream genes, such as VEGFA, PDGFA, and cancer stem cell transcription factors, and promoted tumor angiogenesis and cancer stemness in RCC cells both in vitro and in tumor xenografts. Overall, these findings suggest that HOTAIR and GLI2 could be novel therapeutic targets against RCC. [ABSTRACT FROM AUTHOR]