Diabetes is a global health problem, with diabetic wounds constituting one of its most severe complications. Advanced glycation end products (AGEs) and their receptor, the receptor for advanced glycation end products (RAGE), play a key role in the pathogenesis of diabetic wounds. Accumulated AGEs bind to RAGE, activating various inflammatory and oxidative stress pathways such as NF-κB, PI3K-AKT, and JAK–STAT signaling, impairing normal wound healing. This review describes mechanisms by which the AGEs-RAGE axis disrupts vascular function, immune regulation, and cellular regeneration, thereby driving the formation of chronic non-healing wounds. Furthermore, we discuss emerging therapeutic strategies targeting the AGEs-RAGE axis, such as selective RAGE inhibitors, monoclonal antibodies, gene-based interventions, and AGE scavengers, highlighting their potential to enhance the treatment of diabetic chronic wounds.

Perineural invasion (PNI) is an important factor leading to the recurrence of pancreatic cancer (PanCa). The NGF-TrkA pathway is related to PNI progression. Ganoderma spore lipid (GSL) is a drug with anti-cancer properties. In this study, we find out whether GSL can prevent PNI of PanCa by inhibiting NGF-TrkA pathway. In vitro, wound healing assays, transwell-based assays and three-dimensional tumor-nerve cell co-culture system showed that GSL significantly inhibited the migration and invasion capacity of PanCa cells. Inhibiting the NGF-TrkA pathway is considered an effective approach for treating PanCa. We showed that GSL effectively inhibited NGF-TrkA pathway via immunofluorescence assays and western blotting analysis. The supplement of recombinant NGF reversed the GSL inhibitory effect on the migration and invasion of PANC-1. In vivo, a sciatic nerve invasion animal model was constructed using BALB/c mice. GSL significantly suppressed tumor growth and suppressed the expression of TrkA, NGF, and vimentin and upregulated the level of the epithelial marker E-Cadherin. Moreover, GSL reduced the expression of S100 and PGP9.5. These findings suggested that GSL effectively inhibited the PNI of PanCa cells by downregulating the NGF-TrkA pathway, which may provide a new adjuvant for PanCa treatment.

Direct reprogramming has garnered considerable attention due to its capacity to directly convert differentiated cells into desired cells. Fibroblasts are frequently employed in reprogramming studies due to their abundance and accessibility. However, they are also the key drivers in the progression of fibrosis, a pathological condition characterized by excessive extracellular matrix deposition and tissue scarring. Furthermore, the initial stage of reprogramming typically involves deactivating fibrotic pathways. Hence, direct reprogramming offers a valuable method to regenerate target cells for tissue repair while simultaneously reducing fibrotic tendencies. Understanding the link between reprogramming and fibrosis could help develop effective strategies to treat damaged tissue with a potential risk of fibrosis. This review summarizes the advances in direct reprogramming and reveals their anti-fibrosis effects in various organs such as the heart, liver, and skin. Furthermore, we dissect the mechanisms of reprogramming influenced by fibrotic molecules including TGF-β signaling, mechanical signaling, inflammation signaling, epigenetic modifiers, and metabolic regulators. Innovative methods for fibroblast reprogramming like small molecules, CRISPRa, modified mRNA, and the challenges of cellular heterogeneity and senescence faced by in vivo direct reprogramming, are also discussed.

Phenotypically unstable Schwann cell-like cells (SCLCs), derived from mesenchymal stem cells (MSCs) require intercellular contact-mediated cues for Schwann cell (SCs)-fate commitment. Although rat dorsal root ganglion (DRG) neurons provide contact-mediated signals for the conversion of SCLCs into fate-committed SCs, the use of animal cells is clinically unacceptable. To overcome this problem, we previously acquired human induced pluripotent stem cell-derived sensory neurons (hiPSC-dSNs) as surrogates of rat DRG neurons that committed rat bone marrow SCLCs to the SC fate. In this study, we explored whether hiPSC-dSNs could mimic rat DRG neuron effects to obtain fate-committed SCs from hBMSC-derived SCLCs. hiPSCs were induced into hiPSC-dSNs using a specific chemical small molecule combination. hBMSCs were induced into hBMSC-derived SCLCs in a specific culture medium and then co-cultured with hiPSC-dSNs to generate SCs. The identity of hBMSC-derived SCs (hBMSC-dSCs) was examined by immunofluorescence, western bolt, electronic microscopy, and RNA-seq. Immunofluorescence was also used to detect the myelination capacity. Enzyme-linked immunosorbent assay and neurite outgrowth analysis were used to test the secretion of neurotrophic factors. The hBMSC-dSCs exhibited bi-/tri-polar morphology of SCs and maintained the expression of the SC markers S100, p75NTR, p0, GFAP, and Sox10, even after withdrawing the glia-inducing factors or hiPSC-dSNs. Electronic microscopy and RNA-seq analysis provided evidence that hBMSC-dSCs were similar to the original human SCs in terms of their function and a variety of characteristics. Furthermore, these cells formed MBP-positive segments and secreted neurotrophic factors to facilitate the neurite outgrowth of Neuro2A. These results demonstrated that phenotypically stable and functionally mature hBMSC-dSCs were generated efficiently via the co-culture of hiPSC-dSNs and hBMSC-derived SCLCs. Our findings may provide a promising protocol through which stable and fully developed hBMSC-dSCs can be used for transplantation to regenerate myelin sheath.

2036 Specialty-Gaoqing recipe (2036S-GQR) is a nutraceutical that content docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), grape seed proanthocyanin extract, and bilberry anthocyanins extract. All these ingredients have been proven having various health benefit and 2036S-GQR has been widely and long use in China. However, the toxicity of long-term administration has not yet been reported. The present study explores the long-term toxicity of 2036S-GQR in Wistar rats following the OECD 407 and 408 guidelines. Three dose group was orally administered 2036S-GQR 0.6, 1.2, 2.0 g/kg body weight six day per week for 26 weeks respectively, whereas control group provided distilled water. The investigation of food intake, body weight, hematological, biochemical, and histopathological data was performed in 12, 26 weeks and at the end of recovery period (30 weeks) which was used to assess the toxic effects. The result displays no evident sign of toxicity, no clinically significant change emerges in all measuring data and observation in all dose group, demonstrating 2036S-GQR is harmless to the body under the arranged dose.

Abstract Background A triplet chemotherapy regimen of docetaxel, cisplatin, and 5-fluorouracil (TPF) is used to treat head and neck squamous cell carcinoma; however, it is toxic to bone marrow mesenchymal stem cells (BMSCs). We previously demonstrated that Ganoderma spore lipid (GSL) protect BMSCs against cyclophosphamide toxicity. In this study, we investigated the protective effects of GSL against TPF-induced BMSCs and hematopoietic damage. Methods BMSCs and C57BL/6 mice were divided into control, TPF, co-treatment (simultaneously treated with GSL and TPF for 2 days), and pre-treatment (treated with GSL for 7 days before 2 days of TPF treatment) groups. In vitro, morphology, phenotype, proliferation, senescence, apoptosis, reactive oxygen species (ROS), and differentiation of BMSCs were evaluated. In vivo, peripheral platelets (PLTs) and white blood cells (WBCs) from mouse venous blood were quantified. Bone marrow cells were isolated for hematopoietic colony-forming examination. Results In vitro, GSL significantly alleviated TPF-induced damage to BMSCs compared with the TPF group, recovering their morphology, phenotype, proliferation, and differentiation capacity (p < 0.05). Annexin V/PI and senescence-associated β-galactosidase staining showed that GSL inhibited apoptosis and delayed senescence in TPF-treated BMSCs (p < 0.05). GSL downregulated the expression of caspase-3 and reduced ROS formation (p < 0.05). In vivo, GSL restored the number of peripheral PLTs and WBCs and protected the colony-forming capacity of bone marrow cells (p < 0.05). Conclusions GSL efficiently protected BMSCs from damage caused by TPF and recovered hematopoiesis.

The in vitro derivation of Schwann cells from human bone marrow stromal cells (hBMSCs) opens avenues for autologous transplantation to achieve remyelination therapy for post-traumatic neural regeneration. Towards this end, we exploited human induced pluripotent stem-cell-derived sensory neurons to direct Schwann-cell-like cells derived from among the hBMSC-neurosphere cells into lineage-committed Schwann cells (hBMSC-dSCs). These cells were seeded into synthetic conduits for bridging critical gaps in a rat model of sciatic nerve injury. With improvement in gait by 12-week post-bridging, evoked signals were also detectable across the bridged nerve. Confocal microscopy revealed axially aligned axons in association with MBP-positive myelin layers across the bridge in contrast to null in non-seeded controls. Myelinating hBMSC-dSCs within the conduit were positive for both MBP and human nucleus marker HuN. We then implanted hBMSC-dSCs into the contused thoracic cord of rats. By 12-week post-implantation, significant improvement in hindlimb motor function was detectable if chondroitinase ABC was co-delivered to the injured site; such cord segments showed axons myelinated by hBMSC-dSCs. Results support translation into a protocol by which lineage-committed hBMSC-dSCs become available for motor function recovery after traumatic injury to both peripheral and central nervous systems.

BACKGROUND:It has been observed that bone marrow-derived mesenchymal stem cells (MSCs) migrate towards the injured spinal cord and promote functional recovery when systemically transplanted into the traumatized spinal cord. However, the mechanisms underlying their migration to the spinal cord remain poorly understood.METHODS:In this study, we systemically transplanted GFP- and luciferase-expressing MSCs into rat models of spinal cord injury and examined the role of the stromal cell-derived factor 1 (SDF-1)/CXCR4 axis in regulating the migration of transplanted MSCs to the spinal cord. After intravenous injection, MSCs migrated to the injured spinal cord where the expression of SDF-1 was increased. Spinal cord recruitment of MSCs was blocked by pre-incubation with an inhibitor of CXCR4. Their presence correlated with morphological and functional recovery. In vitro, SDF-1 or cerebrospinal fluid (CSF) collected from SCI rats promoted a dose-dependent migration of MSCs in culture, which was blocked by an inhibitor of CXCR4 or SDF-1 antibody.RESULTS AND CONCLUSION:The study suggests that SDF-1/CXCR4 interactions recruit exogenous MSCs to injured spinal cord tissues and may enhance neural regeneration. Modulation of the homing capacity may be instrumental in harnessing the therapeutic potential of MSCs.

Senescence is a form of durable cell cycle arrest elicited in response to a wide range of stimuli. Senescent cells remain metabolically active and secrete a variety of factors collectively termed senescence-associated secretory phenotype (SASP). SASP is highly pleiotropic and can impact numerous biological processes in which it has both beneficial and deleterious roles. The underlying mechanisms by which SASP exerts its pleiotropic influence remain largely unknown. SASP serves as an environmental factor, which regulates stem cell differentiation and alters its routine. The latter can potentially be accomplished through dedifferentiation, transdifferentiation, or reprogramming. Behavioral changes that cells undergo when exposed to SASP are involved in several senescence-associated physiological and pathological phenomena. These findings provide clues for identifying possible interventions to reduce the deleterious effects without interfering in the beneficial outcomes. In this study, we discuss the multifaceted effects of SASP and the changes occurring in cellular states upon exposure to SASP factors.

组织内的间充质干细胞是人体重要的间充质干细胞来源,作为组织间充质干细胞的一种,肺常驻间充质干细胞(LR-MSC)主要位于肺泡微血管壁中,具有与骨髓间充质干细胞相似的特性,但在治疗肺部疾病时的效果较骨髓间充质干细胞更好.特发性肺纤维化(IPF)是一种以局部组织破坏和细胞外基质沉积为特征的不可逆性慢性肺部疾病,在病理条件下,LR-MSC参与IPF的发病过程,其受部分细胞因子的驱动可分化为成纤维细胞、血管内皮细胞,进而促进肺局部组织纤维化和微血管重构,因此深入研究其具体作用机制对IPF的治疗具有重要意义.

Context The traditional medicine 2036 Specialty-Qiangxin recipe (2036S-QXR) has been widely used in China to improve cardiac function, prevent stroke, and strengthen the immune system. However, its long-term toxicity remains unknown. Objective The present study evaluates the long-term toxicity of 2036S-QXR in rats. Materials and methods 2036S-QXR (0.6, 1.2, and 2.4 g/kg body weight per day) was orally administered for 26 weeks to Wistar rats, while the rats in the control group received distilled water. The effects on urinary, hematological, biochemical, and histopathological parameters were investigated during the study period. Results No significant changes in all tested parameters were observed in the 0.6 and 1.2 g/kg groups, compared with the control group (p < 0.05). Higher levels of alanine aminotransferase (46.00 ± 12.85 vs. 25.40 ± 3.36) and aspartate aminotransferase (152.40 ± 32.52 vs. 111.40 ± 18.78) were observed after 13 weeks in the female rats in the 2.4 g/kg group compared with the control group (p < 0.05), but these returned to the control levels after the recovery period (p > 0.05). Several cases displayed the presence of urine protein (3/7 males and 3/7 females) and mild lesions in the kidney (10/20) and thymus (5/20) in the 2.4 g/kg group, without significant changes compared with the control group (p > 0.05). Discussion and conclusions The present study shows that 2036S-QXR does not cause long-term toxicity, supporting its therapeutic use. To further determine the optimal doses, future studies should test more doses and include more animals in each group.

Skin repair and reconstruction are important after severe wound and trauma. Keratinocyte stem cells (KSCs) in the basal layer of the epidermis can regrow the stratified epidermis but are almost depleted after skin injury. Thus, generating enough KSCs is indispensable for skin regeneration. Pluripotent stem cells such as ESC and iPSC can differentiate into KSCs, but their applications are challenged by ethical issues and risks of tumor formation. Lineage reprogramming from one cell type into another one makes it feasible to generate the desired cell type. Here, we develop a method to convert human fibroblasts into induced keratinocyte stem-like cells (iKSC) by coupling transient expression of reprogramming factors with a chemically defined culture medium, without the formation of iPSC. iKSC resemble normal KSC in the morphological and phenotypic features and can differentiate in vitro and regenerate stratified epidermis after transplantation in vivo. Therefore, iKSC may provide abundant cellular sources for skin repair and regeneration.

Motor neuron loss or degeneration is the typical characteristic of amyotrophic lateral sclerosis (ALS), which often leads to weakness, paralysis, or even death. The underlying mechanisms of motor neuron degeneration and ALS progression remain elusive, and there is no effective treatment for ALS. The advances of stem cells and reprogramming techniques has made it possible to generate patient-specific motor neurons as cell models for studying disease mechanisms and drug discovery. This review comprehensively discusses recent approaches to generate motor neurons from stem cells and somatic cells and highlights the application of induced motor neurons to modeling ALS diseases, dissecting the pathogenesis, and screening new drugs. New perspectives are also discussed on generating patient-specific motor neuron subtypes that are affected by ALS or creating 3D spinal cord organoid models for better recapitulating and understanding ALS.

Background: Bone marrow-derived mesenchymal stem cells (MSCs) have been shown to migrate to injured spinal cords and promote functional recovery when systemically transplanted into the traumatized spinal cord. However, the the mechanisms underlying their migration to spinal cords are not yet fully understood Methods: In this study, we systemically transplanted GFP- and luciferase-expressing MSCs into the rat models of spinal cord injury and examined the role of the stromal cell-derived factor 1 (SDF-1)/CXCR4 axis in regulating the migration of transplanted MSCs to spinal cords. Results: After intravenous injection, MSCs migrated to the injured spinal cord where the expression of SDF-1 was increased. Spinal cord recruitment of MSCs was blocked by pre-incubation with an inhibitor of CXCR4. Their presence correlated with morphological and functional recovery. In vitro, SDF-1 or cerebrospinal fluid (CSF) collected from SCI rats promoted a dose-dependent migration of MSCs, which was blocked by an inhibitor of CXCR4 or an SDF-1 antibody. Conclusions: The study suggests that SDF-1/CXCR4 interactions recruit exogenous MSCs to injured spinal cord tissue and may enhance neural regeneration. Modulation of homing capacity may be instrumental in harnessing the therapeutic potential of MSCs.

Cancer chemotherapeutic agents are frequently toxic to bone marrow and impair bone marrow functions. It is unclear whether ganoderma spore lipid (GSL) can protect bone marrow cells from the cytotoxicity of chemotherapy. To investigate the protective effects of GSL on bone marrow mesenchymal stem cells (MSCs) and hematopoiesis, we examined the effects of GSL on MSCs in vitro and hematopoiesis in vivo after treatment with the chemotherapeutic agent cyclophosphamide. MSCs and peripheral blood cells were isolated and counted from the bone marrow of normal mice were pre-treated with GSL before CTX treatment or co-treated with GSL and CTX, followed by examining the changes in phenotype, morphology, proliferation, apoptosis, and differentiation potentials. The results showed that GSL could reduce the CTX-induced changes in the phenotype of MSCs and maintain the elongated fibroblast-like morphology. MTT and annexin V/propidium iodide (PI) analyses found that GSL pre-treatment and co-treatment increased the proliferation and decreased the apoptosis in CTX-treated MSCs. Furthermore, GSL improved the osteogenic and adipogenic differentiation potentials of CTX-treated MSCs. In vivo, GSL treatment increased the number of peripheral blood cells including white blood cells (WBC) and platelets (PLT) in the CTX-treated mice and enhanced the in vitro formation of hematopoietic lineage colonies (erythrocyte colony forming unit, CFU-E; erythroid burst-forming units, BFU-E; and granulocyte macrophage colony-forming units, CFU-GM) from bone marrow cells in these mice. These findings suggest GSL could protect MSCs and hematopoiesis from the cytotoxicity of CTX and might become an effective adjuvant to attenuate side effects of chemotherapy during cancer treatment.

The aging of stem cells impairs the homeostasis in the tissue. Aged stem cells lose their ability to repair tissue and cause the age-related diseases. Senescent microenvironment is one of the important factors which lead to organism aging. The senescence-associated secretory phenotype (SASP) is an important part of senescent microenvironment. The SASP affects the ability of stem cells to repair and drives the process of aging. Extracellular vesicles (EVs) are thought to play an important role in senescent microenvironment. EVs secreted by senescent cells carry non-coding RNA such as mi RNAs and a variety of active molecules including SASP, which are involved in the regulation of senescent microenvironment. This paper reviews the reasons of the stem cells aging and the research progress of senescent microenvironment, so as to provide the experimental basis and theoretical basis for the clinical application of stem cells.

The term ‘sweat gland regeneration’ refers to a new and expanding field in regenerative medicine research that focuses on the development of innovative therapies allowing the body to replace, restore and regenerate damaged or diseased sweat gland cells and tissues. It combines basic scientific theory and technological approaches including dedifferentiation, biomaterials, tissue engineering, stem cell transplantation and the reprogramming of cell and tissue types. Because of its importance for skin reconstitution in patients suffering from chronic wounds and extensive burns, sweat gland regeneration is becoming an rapidly developing field in regenerative medicine.

Our ultimate goal of in vitro derivation of Schwann cells (SCs) from adult bone marrow stromal cells (BMSCs) is such that they may be used autologously to assist post-traumatic nerve regeneration. Existing protocols for derivation of SC-like cells from BMSCs fall short in the stability of the acquired phenotype and the functional capacity to myelinate axons. Our experiments indicated that neuro-ectodermal progenitor cells among the human hBMSCs could be selectively expanded and then induced to differentiate into SC-like cells. Co-culture of the SC-like cells with embryonic dorsal root ganglion neurons facilitated contact-mediated signaling that accomplished the switch to fate-committed SCs. Microarray analysis and in vitro myelination provided evidence that the human BMSC-derived SCs were functionally mature. This was reinforced by repair and myelination phenotypes observable in vivo with the derived SCs seeded into a nerve guide as an implant across a critical gap in a rat model of sciatic nerve injury.

Here we describe the in vitro derivation of sensory neurons for use in effecting fate commitment of Schwann cell-like cells derived from human bone marrow stromal cells (hBMSCs). We adopt a novel combination of small molecules in an 8-day program that induces the differentiation of human induced pluripotent stem cells into sensory neurons. In co-cultures, the derived sensory neurons present contact-dependent cues to direct hBMSC-derived Schwann cell-like cells toward the Schwann cell fate. These derived human Schwann cells survive passaging and cryopreservation, retain marker expression despite withdrawal of glia-inducing medium and neuronal cues, demonstrate capacity for myelination, and therefore promise application in autologous transplantation and re-myelination therapy.

Objective To investigate the effect of ageing on biological function of cardiac stem cells (CSC) under stress of hypoxia.Methods Primitive mouse Sca1 + CSC were isolated and SA-β-Gal staining was used to confirm the ageing features of the 6th passage (P6) Sca1 + CSC.Before and after hypoxic treatment,the proliferation abilities,the mRNA levels of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF)-A/B,the secretive levels of VEGF and PDGF-BB in P1 and P6 Sca1 + CSC were detected and compared.Ischemic models were established by ligation of the coronary artery in 6 young mice and 6 old mice (ischemic group);in addition,6 young mice and 6 old mice were set as sham group.Three days after establishment of ischemic model,the Sca1 + CSC pool in heart was detected through flow cytometry.Results P6 Sca1 + CSC were positively stained by SA-β-Gal with larger volume compared with P1 Sca1 + CSC,while no positive staining was observed in P6 Sca1 + CSC.The proliferation rate of P6 Sca1 + CSC was significantly lower than that of P1 Sca1 + CSC [(18 ± 5)％ vs (103 ± 27)％,P ＜ 0.01)];after hypoxic treatment,the proliferation rate of P1 Sca1 + CSC [(58 ± 11) ％] was significantly reduced (P ＜ 0.05);no obvious change was found in P6 Sca1 + CSC [(26 ± 8) ％] (P ＞ 0.05).The mRNA levels of VEGF,PDGF-A and PDGF-B were significantly increased in P1 Sca1 + CSC after hypoxic treatment,while no significant changes were found in P6 Sca1 + CSC.After hypoxic treatment,the secretive VEGF and PDGF-BB were significantly increased in P1 Sca1 + CSC [(150 ± 37) μg/L vs (18 ± 6) μg/L,(288 ± 37) μg/L vs (35 ± 6) μg/L,P ＜ 0.01],while no significant changes were found in P6 Sca1 + CSC (P ＞ 0.05).In experiment in vivo,the population of Sca1 + CSC in ischemic group was significantly larger than that in sham group [old mice:(13.2 ± 4.2) ％ vs (1.5 ± 0.4％) ％,young mice:(31.7 ± 6.5) ％ vs (5.0 ±0.6)％];the population of Sca1 + CSC in old mice was significantly smaller than that in young mice in both groups (P ＜ 0.05).Conclusion In Sca1 + CSC,the proliferation ability,secretion and mobilization in response to the hypoxic stress are reduced due to ageing.