Autoimmune thyroiditis (AIT) is one of the most common autoimmune diseases and often causes hypothyroidism in patients. As a traditional formulation in my country, Buzhong Yiqi decoction (BZYQD) has significant effects in improving clinical symptoms of AIT and reducing autoantibody titers, but its specific mechanism of action needs to be further explored. The purpose of this study was to explore the effective targets and related mechanisms of Buzhong Yiqi decoction in AIT mice based on transcriptome sequencing technology. Forty NOD.H-2h4 mice were selected and 0.05% NaI was drinking ad libitum for 8 weeks to establish AIT mice, and drug intervention was performed according to groups for 8 weeks. The groups were as follows: control group, Model Group, Buzhong Yiqi Decoction group (9.56 g·kg-1) and Positive control group (Se yeast tablets, 3.033×10-5 g·kg-1), of which Buzhong Yiqi Decoction was the clinical equivalent dose. Thyroid tissues of the Model Group, blank group and Buzhong Yiqi Decoction group were subjected to transcriptome sequencing to analyze the expression of differential genes, and enrichment analysis was carried out. Hematoxylin and eosin staining (HE staining) was used to detect the pathological changes in thyroid tissues, and enzymelinked immunosorbent assay (ELISA) was used to detect the content of serum thyroglobulin antibody (TGAb) to determine the intervention effect of Buzhong Yiqi Decoction; Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was performed based on the transcriptome sequencing results to detect the expression of TLR8, JUN, TICAM2, TIRAP, and IL-1β mRNA in thyroid tissue. According to the transcriptome results, compared with the blank group, there were 327 significantly up-regulated genes and 440 significantly down-regulated genes in the Model Group; compared with the Model Group, there were 502 significantly up-regulated genes and 380 significantly down-regulated genes in the Buzhong Yiqi Decoction group, mainly enriched in immune inflammation and other related pathways including Toll-like receptors. Animal experiments showed that compared with the control group, the model group had obvious lymphocyte infiltration in thyroid tissue under light microscope, a significant increase in inflammatory cells, a significant increase in TGAb content in serum, and a significant increase in TLR8, JUN, TICAM2, TIRAP, IL-1β mRNA expression was observed (P<0.05 or P<0.01). Compared with the Model Group, Buzhong Yiqi Decoction could significantly improve the inflammatory damage of thyroid tissue in AIT mice, reduce the content of TGAb in serum, and down-regulate the expression of TLR8, JUN, TICAM2, TIRAP, IL-1β mRNA (P<0.05 or P<0.01). Buzhong Yiqi Decoction can effectively improve the inflammatory damage of AIT, and inhibiting the abnormal activation of the Toll-like receptor pathway may be one of its intervention mechanisms.

Intrauterine adhesions (IUA), a leading cause of uterine infertility, are characterized by endometrial fibrosis and a loss of functional regeneration. The impairment of endometrial mesenchymal stem/stromal cells (eMSCs) is central to IUA pathogenesis; however, the absence of reliable markers correlating with disease severity and fibrosis has hindered the understanding of their pathological mechanisms and the development of targeted therapies. Here, ALDH1A1 and ALDH1A2 were identified as novel eMSC markers significantly downregulated in IUA patient samples and scar tissue. ALDHHigh eMSCs exhibited superior self-renewal capability, proliferative capacity, decidualization potential, and angiogenic capability, alongside reduced fibrotic characteristics compared to ALDHLow eMSCs. Transplantation of ALDHHigh eMSCs promoted endometrial regeneration in a murine IUA model. Mechanistically, ALDHHigh eMSCs secreted higher levels of ATRA (all-trans retinoic acid), and their pro-regenerative effect was mediated through the ALDH-RA-RAR (RA Receptor) axis. ATRA and E2 (Estradiol) acted synergistically to promote glandular organoid growth and reprogram ALDHLow eMSCs towards a regenerative phenotype. The RE-Exo@HA hydrogel, composed of ATRA and E2 encapsulated in exosomes embedded within a hyaluronic acid hydrogel, effectively facilitated scarless endometrial repair, demonstrated by restored endometrial architecture, increased gland number, reduced fibrosis, and significantly improved pregnancy outcomes in vivo. Our findings establish ALDH1A1/ALDH1A2 as key functional markers of a regenerative eMSC subpopulation depleted in IUA and highlight the ALDH-RA axis as a critical mechanism governing endometrial repair.

Abstract Background Psoriasis is an immune‐driven dermatosis marked by keratinocyte hyperproliferation. GS‐9620, a TLR7 agonist, previously mitigated EV71‐triggered inflammation in mice; here we probe its anti‐psoriatic potential and mechanisms. Methods IMQ‐induced psoriasis‐like mice were treated with GS‐9620 or MTX; severity was tracked by PASI and histology. Skin/spleen cytokines (IL‐1β, IL‐6, IL‐18, HMGB1, TNF‐α) were quantified via ELISA; immune subsets were quantified by flow cytometry. Autophagy proteins (ATG5/12/16 L1) and NLRP3 were assessed by IHC/Western blot. In vitro, M5‐stimulated primary keratinocytes were treated with GS‐9620 ± autophagy modulators, followed by cytokine and protein analyses. Results GS‐9620 markedly reduced erythema, scaling and epidermal thickness, lowered skin and systemic cytokines, and decreased splenic CD3 + /CD4 + IL‐17A + cells. It restored ATG5/12/16 L1 expression while suppressing NLRP3 both in lesions and in M5‐stimulated keratinocytes, leading to diminished IL‐1β, IL‐6, IL‐18, HMGB1 and TNF‐α release. Conclusions GS‐9620 alleviates psoriasis by enhancing autophagy and dampening NLRP3‐mediated inflammation, offering a promising therapeutic avenue.

Despite recent advancements in antifungal therapy, the antifungal armamentarium remains limited compared to antibiotics available for bacterial infections. Developing novel and effective therapeutic strategies is imperative but challenging. One promising approach involves synergizing existing antifungals with complementary agents to enhance efficacy and reduce the required dosages. This study investigates the synergistic effect of isobavachalcone (IBC) and amphotericin B (AmB) against Cryptococcus neoformans in vitro and in vivo, focusing on ferroptosis modulation. Ferroptosis-related markers, including glutathione (GSH), malondialdehyde (MDA), ferrous ions, and reactive oxygen species (ROS), were analyzed in Caenorhabditis elegans model infected with C. neoformans. IBC (4 μg/mL) significantly lowered AmB's MIC from 1 μg/mL to 0.25 μg/mL, indicating a fourfold enhancement in potency. The IBC-AmB combination caused structural damage to C. neoformans, compromising membrane permeability and cell wall integrity. The combination elevated host GSH levels while reducing ferrous ions, MDA, and ROS in the infected C. elegans model. Mechanistically, the treatment upregulated antioxidant/stress response genes (SKN-1, GST-4, GST-5, GPX-1, DAF-16, CNC-11) and antimicrobial peptides (NLP-29). Conversely, the pro-inflammatory pathway gene PMK-1 was downregulated. The IBC-AmB combination not only reduces the MIC of AmB by fourfold but also enhances antifungal efficacy through a multifaceted mechanism that directly targets the fungal pathogen and modulates the host response. This dual action has the potential to reduce the adverse effects of AmB and improve therapeutic outcomes in the treatment of cryptococcal infections.

The recent designation of Cryptococcus neoformans as a critical-priority fungal pathogen by the World Health Organization highlights the imperative need for novel antifungal agents with distinct mechanisms of action. This study elucidates the novel ferroptotic pathway underlying C. neoformans-induced cell death in Caenorhabditis elegans and investigates the therapeutic potential of isobavachalcone (IBC) through comprehensive evaluation of core biochemical markers: total glutathione (GSH), malondialdehyde, ferrous iron content, and lipid reactive oxygen species (ROS). Integrated transcriptomic analysis via RNA-seq and subsequent RT-qPCR validation revealed critical gene expression patterns associated with antiferroptotic regulation. Our findings demonstrate that C. neoformans infection initiates ferroptosis in C. elegans through iron-dependent lipid peroxidation cascades. Remarkably, IBC administration conferred significant protection against fungal-induced ferroptosis by restoring redox homeostasis-evidenced by elevated GSH levels, attenuated ROS accumulation, and decreased ferrous iron content. Mechanistic investigations identified IBC-mediated upregulation of SKN-1 and GSH biosynthesis genes, coupled with suppression of GPX-1 activity. These coordinated effects disrupted the iron-ROS amplification loop through modulation of the GSH-GPX-1 axis, ultimately extending host lifespan in C. neoformans-challenged models. Our results position IBC as a ferroptosis inhibitor with dual antioxidant and iron-chelating properties, offering a therapeutic strategy against cryptococcal infections through targeting of evolutionary conserved cell death pathways.

Background Tumor-derived proprotein convertase subtilisin/kexin type 9 (PCSK9) facilitates tumor progression, but the role of immune cell-intrinsic PCSK9 in tumor control remains unclear.Methods Orthotopic models of pancreatic cancer and melanoma in Pcsk9-deficient mice were established and tumor-infiltrating immune cells were analyzed using single-cell RNA sequencing and flow cytometry. The effect of genetic disruptions of PCSK9 on murine CD8+ T cells and human chimeric antigen receptor (CAR)-T cells was evaluated both in vitro and in vivo.Results Ablation of host Pcsk9 remarkably suppressed tumor growth and prolonged the survival of tumor-bearing mice, while tumor cells still express PCSK9. The enhanced tumor suppression in Pcsk9-deficient mice depended on CD8+ T cells. Notably, PCSK9 expression was induced in CD8+ tumor-infiltrating lymphocytes (TILs). Consequently, Pcsk9 ablation potentiated the antitumor capacity of CD8+ T cells, showing increased intratumoral infiltration and improved cytotoxic function, along with higher proportions of both effector-memory precursor exhausted (TPEX) and terminally exhausted (TTEX) CD8+ TILs. Additionally, disruption of PCSK9 in both murine CD8+ T cells and human CAR-T cells, synergistic with PD-1 blockade, promoted tumor suppression.Conclusion These findings indicate that PCSK9 inhibits the antitumor function of CD8+ T cells, suggesting it may be a promising target for enhancing T-cell-based cancer immunotherapy.

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the standard first-line treatment for EGFR-mutant cancer (non-small cell lung cancer [NSCLC]), achieving an objective response rate (ORR) of approximately 60%-70%. However, optimizing their therapeutic efficacy remains a challenge. NSCLC cells express the tumor-specific hypoglycosylated Thomsen-nouvelle (Tn) mucin 1 (MUC1) antigen, making them suitable targets for TnMUC1 chimeric antigen receptor (CAR)-T cell therapy. This study shows that EGFR TKIs enhance the efficacy of TnMUC1 CAR-T cell therapy in EGFR-mutant NSCLC, both in vitro and in vivo. EGFR TKIs upregulate TnMUC1 by reducing MUC1 glycosylation, thereby improving TnMUC1 CAR-T cell recognition and cytotoxicity. Specifically, EGFR TKIs modulate TnMUC1-related glycosyltransferases, with core1-beta1,3-galactosyltransferase 1 (C1GALT1) identified as a key enzyme downregulated by EGFR TKIs, suggesting C1GALT1 as a potential therapeutic target.

Although both pre-clinical and clinical studies show promising outcomes, resulting in rapid growth of clinical trials of MSC-based therapies in recent years, the heterogeneity and therapeutic inconsistency of MSCs have severely hampered their clinical applications. Purifying homogenous MSC populations with enhanced specific functions represents one promising approach. We have demonstrated recently that the CD317+ MSCs have enhanced anti-inflammatory functions and improved therapeutic efficacy and consistency. In the current study, we performed both in vitro and in vivo investigations to delineate whether and how CD317 regulates the immune modulation function of MSCs. Our data here indicate that the CD317 directly contributes to the immune suppression function of MSCs stimulated by TNF-α through up-regulating TSG6 via CD317/lipid-raft/TNFR1 complex. The CD317 stabilizes the TNFR1 complex, resulting in hyper-activation of the NF-κB pathway and up-regulation of TSG6, which confers the therapeutic effects of MSCs on the mouse model of ALI (acute lung injury) and IBD (inflammatory bowel disease). Thus, the CD317 stabilizes TNFR1 and confers the anti-inflammatory functions of MSCs via NF-κB/TSG6 Pathway.

Cryptococcus neoformans has been designated as critical fungal pathogens by the World Health Organization, mainly due to limited treatment options and the prevalence of antifungal resistance. Consequently, the utilization of novel antifungal agents is crucial for the effective treatment of C. neoformans infections. This study exposed that the minimum inhibitory concentration (MIC) of isobavachalcone (IBC) against C. neoformans H99 was 8 µg/mL, and IBC dispersed 48-h mature biofilms by affecting cell viability at 16 µg/mL. The antifungal efficacy of IBC was further validated through microscopic observations using specific dyes and in vitro assays, which confirmed the disruption of cell wall/membrane integrity. RNA-Seq analysis was employed to decipher the effect of IBC on the C. neoformans H99 transcriptomic profiles. Real-time quantitative reverse transcription PCR (RT-qPCR) analysis was performed to validate the transcriptomic data and identify the differentially expressed genes. The results showed that IBC exhibited various mechanisms to impede the growth, biofilm formation, and virulence of C. neoformans H99 by modulating multiple dysregulated pathways related to cell wall/membrane, drug resistance, apoptosis, and mitochondrial homeostasis. The transcriptomic findings were corroborated by the antioxidant analyses, antifungal drug sensitivity, molecular docking, capsule, and melanin assays. In vivo antifungal activity analysis demonstrated that IBC extended the lifespan of C. neoformans-infected Caenorhabditis elegans. Overall, the current study unveiled that IBC targeted multiple pathways simultaneously to inhibit growth significantly, biofilm formation, and virulence, as well as to disperse mature biofilms of C. neoformans H99 and induce cell death.

Mesenchymal stem/stromal cells (MSCs) represent a heterogeneous cell population distributed throughout various tissues, demonstrating remarkable adaptability to microenvironmental cues and holding immense promise for disease treatment. However, the inherent diversity within MSCs often leads to variability in therapeutic outcomes, posing challenges for clinical applications. To address this heterogeneity, purification of MSC subpopulations through marker-based isolation has emerged as a promising approach to ensure consistent therapeutic efficacy. In this review, we discussed the reported markers of MSCs, encompassing those developed through candidate marker strategies and high-throughput approaches, with the aim of explore viable strategies for addressing the heterogeneity of MSCs and illuminate prospective research directions in this field.

Pulmonary hypertension (PH) is characterized by vascular remodeling predominantly driven by a phenotypic switching in pulmonary artery smooth muscle cells (PASMCs). However, the underlying mechanisms for this phenotypic alteration remain incompletely understood. Here, we identified that RNA methyltransferase METTL3 is significantly elevated in the lungs of hypoxic PH (HPH) mice and rats, as well as in the pulmonary arteries (PAs) of HPH rats. Targeted deletion of Mettl3 in smooth muscle cells exacerbated hemodynamic consequences of hypoxia-induced PH and accelerated pulmonary vascular remodeling in vivo. Additionally, the absence of METTL3 markedly induced phenotypic switching in PASMCs in vitro. Mechanistically, METTL3 depletion attenuated m6A modification and hindered the processing of pri-miR-143/145, leading to a downregulation of miR-143-3p and miR-145-5p. Inhibition of hnRNPA2B1, an m6A mediator involved in miRNA maturation, similarly resulted in a significant reduction of miR-143-3p and miR-145-5p. We demonstrated that miR-145-5p targets Krüppel-like factor 4 (KLF4) and miR-143-3p targets fascin actin-bundling protein 1 (FSCN1) in PASMCs. The decrease of miR-145-5p subsequently induced an upregulation of KLF4, which in turn suppressed miR-143/145 transcription, establishing a positive feedback circuit between KLF4 and miR-143/145. This regulatory circuit facilitates the persistent suppression of contractile marker genes, thereby sustaining PASMC phenotypic switch. Collectively, hypoxia-induced upregulation of METTL3, along with m6A mediated regulation of miR-143/145, might serve as a protective mechanism against phenotypic switch of PASMCs. Our results highlight a potential therapeutic strategy targeting m6A modified miR-143/145-KLF4 loop in the treatment of PH.

Background Although both preclinical and clinical studies have shown the great application potential of MSCs (mesenchymal stem/stromal cells) in treating many kinds of diseases, therapeutic inconsistency resulting from cell heterogeneity is the major stumbling block to their clinical applications. Cell population diversity and batch variation in the cell expansion medium are two major inducers of MSC heterogeneity. Methods Cell population diversity was investigated through single-cell RNA sequencing analysis of human MSCs derived from the umbilical cord and expanded with fully chemically defined medium in the current study. Then, the MSC subpopulation with enhanced anti-inflammatory effects was studied in vitro and in vivo. Results Our data showed that MSCs contain different populations with different functions, including subpopulations with enhanced functions of exosome secretion, extracellular matrix modification and responses to stimuli (regeneration and immune response). Among them, CD317 + MSCs have improved differentiation capabilities and enhanced immune suppression activities. Underlying mechanism studies showed that higher levels of TSG6 confer enhanced anti-inflammatory functions of CD317 + MSCs. Conclusions Thus, CD317 + MSCs might be a promising candidate for treating immunological disorder-related diseases.

Background Previously, we have demonstrated that the batch variations of human platelet lysate (conventional MSC expansion medium) induce MSC heterogeneity and therapeutic inconsistency. On the other hand, the MSCs expanded with chemical defined medium have improved therapeutic consistency. Methods In the current study, we studied the MSC subpopulation composition and variation in different types and batches of MSC expansion medium with scRNA-seq analysis. Results MSCs expanded with different batches of media have higher levels of heterogeneity from the perspective of cell subpopulation composition at transcriptome levels and therapeutic inconsistency. The CD317 + subpopulation has enhanced immune suppression activities. And the percentage of CD317 + MSCs within MSCs is tightly correlated with its immune suppression activities, and also contributes to the heterogeneity and therapeutic inconsistency of MSCs. the CD317 + MSCs have increased expression levels of PTX3, which might stabilize the TSG6 protein and improve the therapeutic effects Conclusions Thus, purifying CD317 + MSCs is one efficient strategy to reduce MSC heterogeneity and increase the therapeutic consistency of MSCs.

Aim To explore the effect of gypenosides on proliferation and apoptosis of human gastric cancer cells SGC-7901 and AGS and its mechanism. Methods Different concentrations of gypenosides were cultured with human gastric cancer cells SGC-7901 and AGS. Cell viability assay was used to detect cell proliferation activity, and the IC 50 of two kinds of cells was calculated. Cell clone formation assay was conducted to evaluate the proliferation inhibition rate. Flow cytometry was applied to detect the apoptosis rate. caspase activity assay was performed to detect the activities of caspase-9 and caspase-3 in cells. The expression of apoptosis-related proteins was analyzed by Western blot in human gastric cancer cells treated by gypenosides. Results Cell viability assay and clone assay demonstrated that the proliferation of gastric cancer cells SGC-7901 and AGS was inhibited by gypenosides. Flow cytometry showed that gypenosides could increase cell apoptosis significantly compared with the control group. caspase activity assay indicated that gypenosides could increase the activities of caspase-9 and caspase-3 in cells. Gypenosides induced apoptosis by up-regulating the expression of cleaved-PARP,caspase-9,caspase-3 and down-regulating the expression of Bcl-2 proteins in gastric cancer cells. Conclusions Gypenosides exert antitumor activity properties by inhibiting the proliferation of SGC-7901 and AGS cells and inducing apoptosis by regulating the expression of related proteins involved in apoptosis pathway, such as cleaved-PARP,caspase-9,caspase-3 and Bcl-2.

Human rhinovirus (HRV), the main etiologic agent of the common cold, is responsible for significant morbidity, medical costs, and the loss of productivity in the workplace and school. To prevent the spread of HRV, accurate, low-cost and rapid diagnostics of HRV is crucial for identifying those at-risk for the illness associated with HRV, with the most frequently detected species, including HRV species A (HRV-A) and C (HRV-C). Here, a novel HRV-A and/or HRV-C molecular diagnostic assay that integrates reverse-transcription recombinase polymerase amplification assay (RT-RPA) amplification with CRISPR/Cas12a detection, with the result readout using a fluorescence detector or lateral flow strip (LFS). The established assay could be completed within 50 min without complex instruments and skilled technicians. The limit of detection of the RT-RPA-Cas12a-mediated real-time fluorescence or LFS assay could reach 0.1 copy/mu l, and 0.5 copy/mu l for the end-point fluorescence assay with a UV light illuminator readout, respectively. Meanwhile, the assay demonstrates excellent specificity without cross-reactivity to non-target viruses. Furthermore, they were appraised using 80 clinical samples, and RT-RPA-Cas12a-mediated fluorescence or LFS assay displayed high-accuracy with positive and negative predictive agreement of 96.7%, 95% and 100%, respectively. Taken together, the RT-RPA-Cas12a-mediated assay is a rapid, sensitive, and specific detection tool for routine and on-site detection method for HRV-A and/or HRV-C in-fections, and shows great promise for use in resource-poor or constrained settings.

Human bocavirus (HBoV) 1 is considered an important pathogen that mainly affects infants aged 6-24 months, but preventing viral transmission in resource-limited regions through rapid and affordable on-site diagnosis of individuals with early infection of HBoV1 remains somewhat challenging. Herein, we present a novel faster, lower cost, reliable method for the detection of HBoV1, which integrates a recombinase polymerase amplification (RPA) assay with the CRISPR/Cas12a system, designated the RPA-Cas12a-fluorescence assay. The RPA-Cas12a-fluorescence system can specifically detect target gene levels as low as 0.5 copies of HBoV1 plasmid DNA per microliter within 40 min at 37°C without the need for sophisticated instruments. The method also demonstrates excellent specificity without cross-reactivity to non-target pathogens. Furthermore, the method was appraised using 28 clinical samples, and displayed high accuracy with positive and negative predictive agreement of 90.9% and 100%, respectively. Therefore, our proposed rapid and sensitive HBoV1 detection method, the RPA-Cas12a-fluorescence assay, shows promising potential for early on-site diagnosis of HBoV1 infection in the fields of public health and health care. The established RPA-Cas12a-fluorescence assay is rapid and reliable method for human bocavirus 1 detection. The RPA-Cas12a-fluorescence assay can be completed within 40 min with robust specificity and sensitivity of 0.5 copies/μl.

为了将医疗大数据引进医学教育课堂并培养医学生的大数据分析思维,提高医学生对医学数据分析与建模的了解与掌握,该研究设计构建了医疗数据分析科研教学平台,并于2021年在深圳大学医学部的教学中投入使用.该平台引入了大量的医疗数据资源,增强了课程的实践性,使得课堂理论可以落于实地,激发学生对于医学数据分析与建模的兴趣,为新时代的医学教育增添新活力.

Illicium verum Hook.f. (Chinese star anise), a known Chinese traditional spice, is commonly applied in Chinese cuisine and cooking in Southeast Asia. As a kind of medicinal and edible resource, the fruit of I. verum has attracted great attention for its chemical constituents and physiological activities. In this work, the phytochemical study of the fruits of I. verum led to the isolation and identification of 20 compounds, including 6 new lignans and phenylpropanoids (1-6) and 14 known ones (7-20). Their structures were characterized by extensive analysis of spectroscopic data (IR, UV, high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), one-dimensional (1D) and two-dimensional (1D) NMR), electronic circular dichroism (ECD) calculation, and by comparison with literature data. Meanwhile, all compounds (1-20) were evaluated for their antiviral and antioxidant activities. Especially, compound 7 [(-)-bornyl p-coumarate] showed strong antiviral activities against influenza virus A/Puerto Rico/8/34 H1N1 (PR8) with an IC50 value of 1.74 ± 0.47 μM, which is much better than those of Tamiflu (IC50 = 10.01 ± 0.92 μM) and ribavirin (IC50 = 10.76 ± 1.60 μM). The antiviral activity against PR8 of compound 7 was reported for the first time, which was sufficiently confirmed by cell counting kit 8 (CCK-8), cytopathic effect (CPE) reduction, and immunofluorescence assays. In this study, the discovery of antiviral and antioxidant components from the fruits of I. verum could benefit the further development and utilization of this plant.