The formation of chronic refractory wound (CRW) is the result of the combined effect of multiple factors, among which fibrosis plays a dual role in the pathogenesis. Moderate fibrosis can protect the wound, whereas excessive fibrosis leads to pathological scarring, which aggravates tissue hypoxia and stiffness, thereby impeding healing. Thymic stromal lymphopoietin (TSLP) is an important immunoregulatory cytokine that plays a critical role in allergic diseases. Recent studies have revealed that TSLP is also involved in the fibrotic mechanisms underlying CRWs. TSLP regulates key signaling pathways, including transforming growth factor-beta/Sma- and Mad-related protein (TGF-β/SMAD), wingless-type mice mammary tumour virus integration site family/beta-catenin (Wnt/β-catenin), and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT), thereby affecting fibroblast activation, macrophage polarization, chemokine expression, and lysyl oxidase-mediated collagen cross-linking, ultimately contributing to CRW fibrosis. Future studies should focus on exploring the synergistic effects of TSLP with other profibrotic factors and developing combination therapies to balance fibrosis and tissue repair, thereby providing new strategies for the treatment of CRW fibrosis.
Objective To establish a simple and cost-effective protocol for the directed differentiation of induced pluripotent stem cells (iPSCs) into macrophages (Mϕ). Methods Human umbilical cord blood mononuclear cells were reprogrammed into iPSCs by introducing reprogramming factors with Sendai virus vectors. The iPSCs were then induced to differentiate into Mϕ through an embryoid body-based approach, combined with key cytokines including bone morphogenetic protein 4 (BMP4), macrophage colony-stimulating factor (M-CSF), and interleukin-3 (IL-3). Flow cytometry, real-time quantitative polymerase chain reaction and other methods were used to determine the phenotype, function, and expression of immunogenicity-related molecules of the differentiated Mϕ. Results High-purity CD14+CD11b+ Mϕ with normal phagocytic function and polarization capacity were successfully generated by this protocol, without co-culture with animal-derived feeder cells, hypoxic conditions, or cell sorting at intermediate stages. Immunophenotypic analysis revealed that the immune checkpoint CD276+ cells were reduced after the differentiation of iPSCs into Mϕ, which was inconsistent with the stable expression of CD276 observed during the differentiation of iPSCs into β cells or retinal pigment epithelial cells. Conclusion In this study, high-purity Mϕ were successfully generated using a feeder-free, hypoxia-free, and sorting-free system. Furthermore, the lineage-dependent downregulation of CD276 during Mϕ differentiation was observed for the first time.
Objective To establish a method for isolating and expanding perivascular adipose-derived mesenchymal stem cells (PV-ADSC) from rat aortas and to provide a reliable cellular model for investigating vascular remodeling and the related cardiovascular diseases. Methods Perivascular adipose tissue (PVAT) was aseptically collected from the aortas of 1-month-old Sprague Dawley (SD) rats, then rinsed, minced, digested, filtered, and seeded for culture. Upon reaching 80%-90% confluence, the cells were passaged for expansion, and their morphology and proliferation were continuously monitored. Passage 5 (P5) cells were harvested for flow cytometric analysis of surface CD marker expression, and directed differentiation assays with lineage-specific staining were performed to assess the trilineage (adipogenic, osteogenic, chondrogenic) differentiation potential. Results On day 4 of the primary culture, spindle-shaped, stellate, and polygonal cells were observed migrating from the explant edges. Between day 5 and 6, island-like cell clones emerged, and by day 7 to 8, the colonies had coalesced into a confluent monolayer with swirling or parallel alignment. After being passaged, the cells exhibited a homogeneous fibroblast-like spindle morphology and maintained robust proliferation, with no evident senescence or aberrant differentiation through P8. P5 cells showed high expression of the mesenchymal stem cells (MSC) surface markers CD90 (93.60±0.62)%, CD73 (86.80±2.40)%, CD44 (97.77±0.58)%, and CD29 (98.60±0.30)%, yielding a mean positivity rate of (94.19±5.00)%. In contrast, expressions of the hematopoietic stem cells (HSC) surface markers CD45 (5.42±0.78)%, CD34 (7.15±0.49)%, and CD11b/c (3.74±0.08)% were significantly lower, with a mean rate of only (5.44±1.55)%. Directed differentiation assays confirmed the cells' ability to differentiate into adipocytes, osteoblasts, and chondrocytes, as demonstrated by positive staining with Oil Red O, Alizarin Red, and Alcian Blue, respectively. Conclusion Using the method established in this study, rat aortic PV-ADSC were successfully isolated and expanded. These cells exhibited the morphological and surface marker profiles typical of MSC, possessed robust self-renewal capacity, and retained the potential for trilineage differentiation into adipogenic, osteogenic, and chondrogenic lineages. Collectively, these characteristics establish a reliable cellular model for studying vascular remodeling and the related cardiovascular diseases.
B-cell epitopes are regions on the surface of antigens that can be recognized and bound by B-cell receptors or antibodies. Accurate identification of B-cell epitopes of antigens can accelerate the development of drugs such as antibodies and vaccines. This paper conducts a systematic review of representative methods developed in recent years for predicting both B-cell conformational epitopes (such as SEPPA 3.0, DiscoTope-3.0, BepiPred-3.0, etc.) and linear epitopes (such as DLBEpitope, EpiDope, EpitopeVec, etc.). It presents an introduction to these methods, covering aspects such as model architecture, prediction performance, application scope, and illustrative examples, with the aim of providing reference information for related research fields.
Objective To investigate the role of berberine (BBR) in chronic lymphocytic leukemia (CLL) and to determine whether it exerts anti-tumor effects by directly targeting and inhibiting Lck/Yes tyrosine kinase (Lyn), a novel Src family kinase, thereby inducing leukemia cell apoptosis. Methods Molecular docking was employed to predict the binding potential between BBR and Lyn kinase, and biotin pull-down assay was conducted to validate their direct interaction. In vitro experiments utilized the human chronic B-cell leukemia cell line MEC-1, with cell viability and apoptosis assessed via CCK-8 and TUNEL staining, respectively. Key proteins in the B-cell receptor (BCR) pathway, including Lyn, spleen tyrosine kinase (Syk), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and apoptosis-related markers Bcl2-associated X protein (BAX), Bcl2-associated agonist of cell death (BAD), cleaved caspase-3(c-caspase-3), and B-cell lymphoma 2 (Bcl2), were analyzed by Western blot. Transcriptional levels of downstream genes, including cyclin D1 (Cyclin D1), Bcl2, and myelocytomatosis viral oncogene homolog (c-Myc), were quantified using real-time quantitative PCR. Functional rescue experiments were performed using Lyn-overexpressing lentiviral stable cell lines. In vivo, a C-NKG mouse leukemia model was established via tail vein injection, with tumor infiltration in the spleen, liver, and lungs evaluated by HE staining, and therapeutic effect of BBR assessed by survival analysis. Results In vitro, BBR inhibited MEC-1 cell proliferation in a concentration-dependent manner and induced apoptosis, while suppressing the phosphorylation of BCR pathway proteins and downstream gene expression. Molecular docking and pull-down assays confirmed the direct binding between BBR and Lyn. The overexpression of Lyn reversed BBR-induced apoptosis and pathway inhibition. In vivo, BBR treatment significantly reduced organ infiltration and prolonged survival in leukemic mice, which can be reversed by Lyn overexpression. Conclusion BBR induces CLL cell apoptosis and inhibits tumor progression in vitro and in vivo by directly targeting Lyn kinase and suppressing the BCR-Lyn-PI3K-AKT signaling pathway. These findings provide experimental evidences supporting BBR as a natural Lyn-targeted therapeutic agent for CLL.
Type 3 innate lymphoid cells (ILC3s) are a subset of innate immune cells regulated by the transcription factor retinoic acid receptor-related orphan receptor γt (RORγt). They are primarily distributed in the intestinal mucosa and lymphoid tissues, where they secrete cytokines such as interleukin 17(IL-17), IL-22, and granulocyte-macrophage colony-stimulating factor (GM-CSF), playing a crucial role in maintaining intestinal mucosal homeostasis, defending against pathogen invasion, and modulating immune responses. Renal fibrosis (RF) is a pathological process triggered by pathogenic factors such as trauma, infection, inflammation, or metabolic abnormalities. It is characterized by damage to renal parenchymal cells, abnormal deposition of extracellular matrix (ECM), and progressive fibrosis of renal tissue, ultimately leading to irreversible loss of kidney function. Recent studies have revealed that ILC3s not only contribute to intestinal homeostasis but are also involved in the progression of RF. Therefore, this review summarizes the research advances in the mechanisms of ILC3-mediated cross-organ regulation in RF, aiming to provide new perspectives for the prevention and treatment of RF.
Objective To investigate the effects of Cinobufotalin (Cino) on the tumor microenvironment (TME) of colorectal cancer (CRC) and the chemosensitivity to Oxaliplatin (OX). Methods We explored the clinical expression differences of CD244 in the TCGA-COAD cohort. Through UMAP dimensionality reduction analysis of single-cell sequencing data, we clarified the cellular localization of CD244 and analyzed its interaction characteristics with immune cells. In vivo experiments, a CRC tumor-bearing mouse model was established and randomly divided into 4 groups: Tu group, Cino group, OX group, and Cino combined with OX group. Intraperitoneal injection was performed every other day for treatment. Experimental techniques such as tumor volume measurement, fluorescence signal/mass changes and tissue morphology changes, real-time quantitative PCR, Western blot, and immunofluorescence staining were used to detect tumor proliferation, metastasis, expression distribution of CD244+ macrophages and T cell activity. In vitro experiments, RAW264.7 cells were stimulated with interleukin 4 (IL-4) to construct a M2-type macrophage model. The effects of Cino, OX or Cino combined with OX on the expression of CD244+ macrophages were observed, and the conditioned medium (CM) of the corresponding treated macrophages was used to culture T cells. The expression of T cell exhaustion and activation markers was detected, and the killing function of T cells was tested. Results In vivo experiments, Cino treatment had no significant effect on tumor growth, proliferation and metastasis in mice. OX treatment and its combination with Cino inhibited tumor proliferation and metastasis, with the Cino-OX combination exhibiting an enhanced inhibitory effect. Both Cino and OX inhibited the expression of CD244+ macrophages in tumor tissues, and their combination showed a synergistic inhibitory effect. Neither Cino nor OX alone had significant effect on T cell activity, their combination could inhibit T cell exhaustion and increase CD8+ T cell infiltration. In vitro experiments, Cino and OX treatments inhibited the expression of CD244+ macrophages, with their combination exhibiting an enhanced inhibitory effect; after IL-4 treatment, the supernatant was collected to culture T cells, and the tumor-killing ability of T cells decreased. The CM treated with Cino or OX could enhance the killing ability of T cells, and the combination showed a stronger killing ability. Conclusion Cino enhances the tumor-killing capacity of T cells and reshapes the tumor microenvironment by specifically targeting and inhibiting CD244+ macrophages, thereby increasing the chemosensitivity of CRC to OX.
Objective To investigate the distribution characteristics of the genotypes and allele frequencies of HNA-1~-5 alleles in a blood donor population from Jiangsu Province. Methods Genomic DNA was extracted from 300 peripheral blood samples collected from healthy donors. All samples were genotyped using an optimized polymerase chain reaction-sequence specific primer (PCR-SSP) protocol. Randomly selected results were verified by direct sequencing, and genotype distributions were tested for conformity to Hardy-Weinberg equilibrium. Samples identified as potential HNA-1 null were confirmed by polymerase chain reaction sequencing-based typing (PCR-SBT) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results The allele frequencies for HNA-1a, HNA-1b, and HNA-1c were 58.6%, 41.4%, and 0%, respectively. Two identified HNA-1 null cases were confirmed to result from a complete deletion of the Fc gamma receptor IIIb (FCGR3B) gene. The allele frequency of the HNA-2-encoding CD177 gene was 100%, with no HNA-2 null cases detected due to the homozygous c.787 A>T (TT) mutation. The allele frequencies for HNA-3a, HNA-3b, HNA-4a, HNA-4b, HNA-5a, and HNA-5b were 63.0%, 37.0%, 100%, 0%, 88.8%, and 11.2%, respectively. No statistically significant differences in HNA allele frequencies were found between the Jiangsu population and those from Shanghai or Zhejiang. However, significant differences were observed when compared to the Guangzhou population specifically for the HNA-1a/1a (P=0.008), HNA-3a/3a (P=0.033), and HNA-3b/3b (P=0.022) genotypes. Conclusion This study establishes the HNA genotype and allele frequency profile for the Jiangsu blood donor population, providing essential baseline data for research into HNA-related alloimmunization. Furthermore, it optimizes a methodology suitable for large-scale HNA screening and establishes a comprehensive confirmatory testing protocol for HNA-1 null, thereby laying a technical foundation for subsequent research on this allele.
Human cytomegalovirus (HCMV) infection is one of the most common and severe viral complications following allogeneic hematopoietic stem cell transplantation (allo-HSCT), significantly increasing the risk of graft-versus-host disease (GVHD) and non-relapse mortality. Letermovir, as the first inhibitor targeting the HCMV terminase complex, blocks the cleavage of viral DNA and its packaging into viral capsids, demonstrating remarkable efficacy in early post-transplant HCMV prophylaxis. It significantly reduces the incidence of clinically significant HCMV infection (csCMVi) and indirectly contributes to improved GVHD- and relapse-free survival. However, while the potent viral suppression of this treatment provides clinical benefits, it also limits viral antigen exposure, which leads to delays in the reconstitution of HCMV-specific T cells and NK cells and consquently the increased risk of late-onset HCMV reactivation after drug discontinuation. This may also be associated with an elevated risk of Epstein-Barr virus reactivation and post-transplant lymphoproliferative disease (PTLD). This review aims to systematically summarize the dual effects of letermovir in allo-HSCT and analyzes its pharmacological mechanisms, clinical efficacy, impact on immune reconstitution, and potential consequences. Furthermore, it explores individualized management strategies based on immune monitoring to provide a theoretical basis for optimizing clinical practice.
Objective To explore the immune landscape of patients with lung adenocarcinoma with drainage lymph nodes by single-cell sequencing technology. Methods The lung cancer single cell dataset (GSE277742) was obtained from the Gene Expression Omnibus (GEO) database of the National Center for Biotechnology Information (NCBI). The dataset included intrathoracic DLN from 18 patients with pathologically confirmed lymph node metastasis and 4 control patients without evidence of metastasis. R language was used for data quality control, dimensionality reduction clustering, cell subgroup annotation and cell communication analysis to identify key cell subsets and screen differentially expressed genes. Kaplan-Meier survival analysis was performed using the TCGA database. Results Single cell sequencing analysis showed that the proportion of CD8+ T cells in patients with draining lymph nodes was significantly higher, among which cyclin-dependent kinase 6(CDK6), cyclin D3(CCND3), CCNH, cyclin-dependent kinase inhibitor 2A/B/D(CDKN2A/B/D) and retinoblastoma transcriptional corepressor 1(RB1) were highly expressed. TCGA database analysis showed that the expression of genes related to the BIOCARTA_CELLCYCLE_PATHWAY pathway was significantly different between tumor and normal samples, and was related to survival. Conclusion Several key genes differentially expressed in the draining lymph nodes of patients with lung adenocarcinoma are screened by bioinformatics methods, which provides a new perspective for further analysis of the regulatory mechanism of the immune microenvironment of lung adenocarcinoma with draining lymph nodes.
Malignant tumors are major diseases that threaten human health. A long-standing focus of research is how to eliminate tumor cells while minimizing damage to healthy tissues. This article systematically reviews the dual role of iron metabolism in tumorigenesis and progression. Firstly, disruption of iron homeostasis promotes malignant transformation through mechanisms including oxidative stress, signaling transduction, and epigenetic regulation. Secondly, ferroptosis triggered by iron overload offers a novel avenue for targeted cancer therapy. Based on the established relationship between iron metabolism and cancer progression, this paper proposes therapeutic strategies centered on iron chelators, ferroptosis inducers, and nanomaterials. It further elaborates on recent advances in combining iron metabolism modulation with chemotherapy, immunotherapy, epigenetic therapy, and other modalities. Finally, by elucidating the core mechanism through which iron metabolism promotes cancer, we suggest potential therapeutic directions, aiming to provide new insights for advancing precision oncology and developing novel anticancer agents.
Objective To investigate the heterogeneity characteristics of cancer-associated fibroblasts (CAF) in acral melanoma (AM) and their functional roles in the tumor microenvironment (TME), providing a theoretical basis for developing effective therapeutic strategies. Methods The single-cell RNA sequencing data used in this study were previously generated by our research group and have been deposited in the HRA001804 dataset of the National Genomics Data Center (NGDC). Five AM patient samples were selected from this dataset for analysis, including 4 primary tumor samples (PL1, PL2, PL4, PL5) and 1 lymph node metastasis sample (LG2). Bioinformatics methods including unsupervised clustering, pseudotime trajectory analysis, transcription factor regulatory network analysis, and cell-cell communication analysis were used for data mining. Results Three subtypes of CAF were identified from 41 960 high-quality cells: immunomodulatory CAF (iCAF), myofibroblastic CAF (mCAF), and proliferative CAF (pCAF). Pseudotime trajectory analysis demonstrated that mCAF were located at the initiation of differentiation, while iCAF and pCAF were distributed in distinct terminal branches. FOS-like 1, AP-1 transcription factor subunit (FOSL1) was identified as a key transcription factor regulating the differentiation of CAF into pCAF, and its co-expression modules were enriched in the P53 signal pathway, AP-1 transcription factor network, and MYC-mediated cell proliferation pathways. Cell communication analysis revealed an interaction network centered on iCAF, with critical pathways including CSPG4-(ITGA2+ITGB1), GDF15-TGFBR2, LGALS9-CD45/CD44, and CD70-CD27 signal pathways. Conclusion CAF in AM exhibits high heterogeneity, with FOSL1 playing a key regulatory role in CAF differentiation. CAF participates in tumor microenvironment regulation through complex intercellular communication networks. These findings provide important insights into understanding the biological characteristics of acral melanoma and developing targeted therapeutic strategies.
Objective To elucidate the potential molecular mechanism of baicalein in the treatment of allergic contact dermatitis (ACD). Methods An in vitro inflammatory model was established using human keratinocytes (HaCaT cells) stimulated with tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ). The regulatory effect of baicalein on inflammatory factors was detected by quantitative real time polymerase chain reaction. Network pharmacology was applied to screen the common targets of baicalein and ACD. Key signaling pathways were predicted via a protein protein interaction (PPI) network and pathway enrichment analysis, and the binding capacity of baicalein to core targets was evaluated by molecular docking. Results In vitro experiments showed that baicalein significantly reduced the expression of pro-inflammatory factors in HaCaT cells induced by TNF-α combined with IFN-γ, but had no significant effect on the T helper 1 (Th1)/T helper 17(Th17)cell imbalance, suggesting that its anti inflammatory mechanism was not completely dependent on inflammatory pathways such as T cell activation and Th17 differentiation. A total of 46 potential targets of baicalein for ACD were screened by network pharmacology, and the functional enrichment was associated with inflammatory pathways including T cell activation and Th17 differentiation. Furthermore, 11 core targets were obtained from the 46 potential targets using CytoHubba and MCODE algorithms. Molecular docking results demonstrated that baicalein exhibited high binding affinity to core targets related to the cytochrome P450 (CYP450) family, among which the affinity with CYP1A1 was the highest (binding energy=-10.17 kcal.mol-1, RMSD=1.14 ). Conclusion Baicalein exhibits significant therapeutic potential against ACD. It can downregulate the expression of inflammatory factors in HaCaT cells induced by TNF-α combined with IFN-γ, and its mechanism may be related to the interactions with CYP450 family-related targets.
Inflammatory bowel disease (IBD) belongs to a group of idiopathic intestinal inflammatory disorders that affect the ileum, rectum, and colon. There is still no effective treatment methods, and with the increasing number of patients, IBD has brought a heavy burden to the medical system. Chronic inflammation mediated by abnormal activation of the intestinal mucosal immune system plays an important role in the occurrence and development of IBD. As a common intracellular sensor that activates the innate immune system, inflammasomes have a signaling cascade regulated by ubiquitination/deubiquitination. Multiple studies have confirmed that NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is continuously activated in the progression of IBD, therefore targeting this oligomeric protein complex is of great significance for the treatment of IBD. This review systematically introduces the activation mechanism of NLRP3 inflammasome, the regulation of NLRP3 signaling cascade by ubiquitination/deubiquitination modification, and the role of NLRP3 inflammasome in the progression of IBD under the post-translational modification regulation. It also systematically discusses the feasibility of targeting this protein complex to provide new ideas for the treatment of IBD.
The neutrophil-to-lymphocyte ratio (NLR), as a simple and reliable biomarker of systemic inflammation, has garnered significant attention in the clinical management of various inflammatory diseases and malignancies. By quantifying the relative counts of neutrophils and lymphocytes, this parameter reflects the body's immune-inflammatory status. This article provides a comprehensive review of the current research on the role of NLR in chronic obstructive pulmonary disease (COPD) concomitant with advanced lung cancer (LC) and aims to offer insights into novel diagnostic approaches, prognostic evaluation strategies, and potential therapeutic implications for clinical practice.
Objective To express and purify Alicyclobacillus acidiphilus Cas12b (AaCas12b) protein, and establish a rapid clustered regularly interspaced short palindromic repeats (CRISPR)-based assay for Mycobacterium tuberculosis (Mtb) detection, thereby providing a novel tool for clinical diagnosis of tuberculosis (TB). Methods Following construction and transformation of the recombinant expression vector into E. coli BL21 (DE3), soluble expression of the recombinant AaCas12b protein was induced with low-concentration isopropyl β-D-thiogalactopyranoside (IPTG) at low temperature, followed by purification using nickel-affinity chromatography. Subsequently, a specific recombinase polymerase amplification (RPA) assay targeting the IS6110 insertion sequence of Mtb was developed, and the corresponding single guide RNA (sgRNA) was prepared via in vitro transcription. Then a novel CRISPR-AaCas12b detection assay for Mtb was established, and its diagnostic performance was systematically evaluated against conventional clinical methods including acid-fast staining, T-SPOT.TB assay, Mycobacterium culture, and GeneXpert assay. Results A prokaryotic expression and purification system for recombinant AaCas12b was successfully established, yielding AaCas12b protein with activity at a concentration of 13.42 mg/mL and a recovery rate of 61.45%. A specific RPA-CRISPR-AaCas12b detection system was established targeting the IS6110 sequence, with a limit of detection (LOD) of 1.5 CFU/mL for Mtb. In clinical sample validation, the system achieved a sensitivity of 95.83% (95% CI: 79.97%~99.27%), a specificity of 92.31% (95% CI: 75.88%~97.88%), and an accuracy of 94.00% (95% CI: 83.78%~97.93%). Conclusion The established AaCas12b protein preparation system is efficient and stable, enabling the production of low-cost, high-activity tool enzymes. The RPA-CRISPR-AaCas12b TB diagnostic assay targeting the IS6110 sequence exhibits excellent specificity and sensitivity, providing a technical reference and experimental basis for the further development of rapid TB diagnostic platforms.
Objective To investigate the effect of histone deacetylase 11 (HDAC11) on oxidized low density lipoprotein (ox-LDL)-induced mouse macrophage foam cell formation. Methods Macrophages derived from the RAW264.7 cell line were divided into three groups: Control group, ox-LDL group, and ox-LDL combined with Rapamycin group. Oil red O staining was used to observe the formation of foam cells, the content of total cholesterol (TC) and triglyceride (TG) in macrophages were measured using microplate reader. The expression of HDAC11, microtubule-associated protein 1 light chain 3-β (LC3B) II and p62 were detected by Western blot assay. Si-HDAC11 was transfected into macrophages, and qRT-PCR and Western blot assay were employed to measure the interference efficiency of HDAC11. On the basis of ox-LDL treatment, the expression of HDAC11 in RAW264.7 cells was knock down. Oil red O staining was used to observe the lipid droplet content, the content of TC and TG were detected by using microplate reader, and LC3BII and p62 protein expressions were detected by Western blot assay. Results Compared to the control group, the protein expression of LC3BII was reduced, while p62 and HDAC11 protein were increased, which is accompanied by a significant increase of the lipid droplet content, TC and TG. Simultaneously, treated RAW264.7 cells with Rapamycin, an autophagy agonist, LC3BII and p62 protein expressions, as well as lipid droplets, TC, and TG content changes were significantly alleviated in RAW264.7 cells. LC3BII expression was significantly increased after the transfection of si-HDAC11 into macrophages, while p62 protein expression was decreased, and intracellular lipid droplets, TC and TG contents were also reduced correspondingly. Conclusion HDAC11 promotes ox-LDL-induced mouse macrophage foam cell formation via inhibiting autophagy.
Sepsis is one of the major diseases threatening human health. Under inflammatory stress, the rapid activation of glucocorticoids (GCs) and catecholamines (CAs) in the adrenal gland is crucial for maintaining the body's homeostasis. During severe inflammation, impairment of the hypothalamic-pituitary-adrenal (HPA) axis leads to suppressed secretion of corticotropin-releasing hormone (CRH) and adrenocorticotropic hormone (ACHT). However, studies reveal that the systemic glucocorticoid levels can remain normal or even elevated, suggesting a transition in the stress response mechanism from the central, pituitary-dependent regulatory system to local, pituitary-independent regulatory mechanisms. Among the various potential mechanisms, activation of the local adrenal microenvironment plays a significant role. In this microenvironment, interactions between adrenal cortical cells, medullary cells, vascular endothelial cells, and various immune cells contribute to the sustained production of GCs, which is pivotal in maintaining the internal homeostasis. This review focuses on the role of the local adrenal microenvironment in regulating the secretion of GCs and CAs, key hormones for maintaining homeostasis under inflammatory stress, with the aim of providing new insights into the etiology and pathogenesis of adrenal diseases.
Following Mycobacterium tuberculosis (Mtb) infection, the pathogen can establish a metabolically quiescent or replication-arrested dormant state within the host, leading to asymptomatic latent tuberculosis infection (LTBI). Under specific conditions, these dormant bacilli may reactivate and replicate, resulting in symptomatic clinical infection and constituting a major reservoir for active tuberculosis (TB). Dormant Mtb exhibits reduced susceptibility to conventional anti-TB drugs, and current TB vaccines fail to prevent the establishment or reactivation of dormant Mtb, rendering latent infection a major challenge in TB control. The formation of Mtb dormancy requires a unique microenvironment characterized by hypoxia, nutrient deprivation, and host immunosuppression. As an intracellular pathogen, Mtb primarily infects macrophages. Recent studies have revealed that Mtb can also infect mesenchymal stem cells (MSCs), where it persists in a dormant state for a long term. MSCs exhibit a microenvironment marked by hypoxia, cellular quiescence, lipid droplet biogenesis, and elevated autophagic activity. Moreover, MSCs exert immunosuppressive effects on the host immune response to Mtb, thereby facilitating the formation of bacterial dormancy and long-term intracellular survival, which further contributes to TB reactivation and relapse. This review summarizes current advances in understanding Mtb infection of MSCs and the underlying mechanisms associated with Mtb dormancy within this cellular niche.
Objective Exploring the role of transcription factor EB (TFEB)-mediated nuclear translocation in lysosomal degradation of glutathione peroxidase 4 (GPX4) in ferroptosis of human embryonic trophoblast cells HTR8-S/Vneo. Methods HTR8-S/Vneo cells were divided into the following groups: normal group, hypoxia group, hypoxia+si-NC group (cells transfected with si-NC), hypoxia+si-TFEB group (cells transfected with si-TFEB), hypoxia+si-TFEB+RSL3 group (cells transfected with si-TFEB and treated with the ferroptosis inducer RSL3), and hypoxia+si-TFEB+PP242 group (cells transfected with si-TFEB and treated with the lysosome activator PP242). The viability, invasion number, and migration rate of cells were assessed by the CCK-8 kit, Transwell assay, and wound healing assay, respectively. The content of Fe2+ in cells was detected using the FerroOrange probe. ROS levels were measured using the DCFH-DA reactive oxygen species (ROS) fluorescent probe. Enzyme-linked immunosorbent assay (ELISA) was employed to determine the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), glutathione (GSH), and the activity of superoxide dismutase (SOD). Western blot analysis was performed to examine the protein expression of TFEB, phosphorylated TFEB (p-TFEB), GPX4, and TFEB nuclear translocation. The lysosomal fluorescent intensity in cells was assessed using a lysosomal green fluorescent probe. Results Compared with the normoxia group, the hypoxia group exhibited significantly reduced viability, invasion number, and migration rate of cells. Intracellular Fe2+ levels, ROS fluorescence intensity, LDH release, and MDA level were markedly increased, while SOD activity and GSH levels were significantly decreased. TFEB and p-TFEB protein expression showed significant up-regulation, whereas GPX4 protein expression was notably down-regulated. TFEB nuclear translocation occurred with enhanced lysosomal fluorescence intensity. Compared with the hypoxia+si-NC group, the hypoxia+si-TFEB group demonstrated significantly increased viability, invasion number, and migration rate of cells, accompanied by reduced ferroptosis levels. TFEB nuclear translocation was inhibited, and lysosomal fluorescence intensity decreased. Compared with the hypoxia+si-TFEB group, ferroptosis-related indicators in HTR8-S/Vneo cells were reversed in the hypoxia+si-TFEB+RSL3 group. In contrast, the cellular ferroptosis level was elevated in the hypoxia+si-TFEB+PP242 group. Conclusion Inhibition of TFEB nuclear translocation reduces ferroptosis in human embryonic trophoblast cells HTR8-S/Vneo, possibly by suppressing the lysosomal degradation of GPX4.
