High-altitude chronic hypoxia can alter the immune status, yet the immune regulatory characteristics of various vaccine adjuvants in this environment remain to be fully elucidated. Accordingly, emphasis is placed on evaluating the regulatory effects of chronic hypoxia on specific humoral, cellular, and memory T cell responses elicited by different adjuvants.

Using ovalbumin (OVA) as the antigen, Th2-biased adjuvant aluminum hydroxide (Alum), Th1-biased adjuvant CpG, and Th1/Th2-balanced adjuvant QML were combined for intramuscular immunization in mice on days 0 and 14. Th1-biased adjuvant CpG, or Th1/Th2-balanced adjuvant QML for intramuscular immunization on day 0 and 14 (n=16). A total of 160 SPF female C57BL/6 mice (6 to 8 weeks old, weighing 16 to 18 g) were randomly divided into 5 groups (n=32): PBS group (sterile PBS), OVA group (5 μg OVA), OVA+Alum group (5 μg OVA and 100 μg Alum), OVA+CpG group (5 μg OVA and 20 μg CpG), and OVA+QML group (5 μg OVA and 50 μL QML). Each group was further divided into normoxia control (n=16, housed in 21% O₂ environment) and chronic hypoxia (n=16, simulated altitude 5 800 m) subgroups. On days 14 and 60 after the last immunization, the serum levels of specific IgG, IgG1, and IgG2c antibodies were measured by ELISA; enzyme-linked immunospot assay (ELISpot) was used to detect the numbers of cells secreting IFN-γ, IL-4, and IL-17A in the splenocytes. On day 14 after the last immunization, flow cytometry was performed to determine the proportions of CD4⁺IFN-γ⁺, CD4⁺IL-4⁺, CD4⁺IL-17A⁺, CD8⁺IFN-γ⁺, and CD8⁺IL-2⁺ cell subsets in the spleen.

Chronic hypoxia significantly inhibited the ability of OVA alone to induce IgG antibodies (P<0.01), and all 3 adjuvants reversed this effect. QML exerted a stable and superior enhancing effect on serum-specific IgG, IgG1, and IgG2c levels compared to CpG (P<0.05). Notably, CpG induced significantly higher levels of total IgG (P<0.01) and IgG1 (P<0.05) in the chronic hypoxia group than in the corresponding normoxia group at days 60 after the final immunization. For cellular immunity, QML stably increased the proportions of CD4⁺IFN-γ⁺ and CD4⁺IL-17A⁺ cells, as well as the secretion levels of IFN-γ and IL-4, under both normoxic and hypoxic conditions. In contrast, the T cell regulatory effects of CpG were hypoxia-dependent, with significant enhancement of IFN-γ-secreting cell numbers in the hypoxia group at both short-term (14 d, P<0.01) and long-term (60 d, P<0.05) time points. Analysis of memory T cells showed that all 3 adjuvants maintained CD4⁺Tcm levels under hypoxia. Additionally, CpG increased the proportions of CD8⁺Tcm (P<0.05) and CD8⁺Tem (P<0.01) in the hypoxic group, while Alum only specifically elevated CD4⁺Tem levels under hypoxic conditions (P<0.05).

QML stably induces comprehensive immune responses under both normoxic and hypoxic conditions, while CpG exhibits advantages in Th1-type response enhancement and CD8⁺ memory T cell expansion under chronic hypoxia, providing experimental evidence for adjuvant selection in chronic hypoxic environments such as high-altitude regions.

Helicobacter pylori (Hp) infection is prevalent worldwide and represents a risk factor for various chronic inflammatory and neoplastic gastric diseases. In recent years, lipid nanoparticles (LNPs) have demonstrated broad application prospects in vaccine adjuvant research owing to their favorable delivery capacity and immunomodulatory potential. Preparation of LNPs and evaluation of their immunoenhancing effects as adjuvants for a Helicobacter pylori recombinant protein vaccine, as well as their protective efficacy against H. pylori infection.

LNPs were prepared using microfluidic technology, with morphology examined by transmission electron microscopy and particle size and polydispersity index characterized by dynamic light scattering. The activating effects of LNPs on bone marrow-derived dendritic cells (BMDCs) were evaluated in vitro. For in vivo experiments, female BALB/c mice (6 to 8 weeks old, weighing 17 to 20 g) were randomly divided into (n=10): PBS group, UreB/NapA group, and LNP-H+UreB/NapA group. Mice were immunized by intramuscular injection on days 0, 14, and 28. Serum specific antibody titers were measured by ELISA at day 14 after the final immunization, and IFN-γ and IL-17A secretion in mouse spleens was detected by ELISpot. Mice were challenged with H. pylori to establish an infection model. At 4 weeks post-infection, H. pylori colonization in gastric tissue was quantified by probe-based real-time PCR, and adhesion inhibition ability of immunized mice against H. pylori was evaluated through in vitro AGS cell adhesion assay.

The prepared LNPs exhibited uniform morphology, with mean particle size of 100 to 120 nm and polydispersity index of 0.1 to 0.2. LNPs promoted the phagocytosis of antigens by BMDCs and significantly increase the expression levels of costimulatory molecules CD80, CD86, and CD40 on BMDC surfaces (P<0.001), indicating potential to promote BMDC maturation. LNPs also significantly enhanced the secretions of IL-6 and IL-1β in the supernatants of BMDCs (P<0.0001). When combined with UreB and NapA, LNPs remarkably elevated the serum titers of specific total IgG, IgG1, and IgG2a (P<0.05), and promoted the secretions of specific IFN-γ and IL-17A by splenic lymphocytes (P<0.05), suggesting that LNPs strengthen specific T-cell immune responses. Furthermore, LNPs reduced the colonization of H. pylori in mouse gastric tissues and significantly inhibited the adhesion to gastric mucosal epithelial cells.

LNPs can serve as an effective adjuvant for H. pylori recombinant protein vaccines, effectively enhancing humoral and cellular immune responses and conferring protective effects against H. pylori infection.

The escalating threat of antimicrobial resistance poses significant clinical challenges, particularly in cases of co-infections involving Staphylococcus aureus (SA) and Pseudomonas aeruginosa (PA), which synergistically increase disease severity and complicate therapeutic interventions. A bivalent nanovaccine, MntC-rePO@LS, was constructed by co-displaying the SA antigen Manganese transporter C (MntC) and the PA recombinant PcrV-OprI fusion antigen (rePO) based on self-assembling lumazine synthase (LS) nanoparticles. We hypothesized that this vaccine would provide superior immunoprotection against both single and co-infections with SA and PA compared to monomeric or physically mixed formulations.

The MntC-rePO@LS bivalent nanoparticles were prepared using genetic engineering and the SpyTag/SpyCatcher system. Female BALB/c mice (6 to 8 weeks old, weighing 18 to 20 g) were randomly divided into 6 groups: PBS, LS (8.57 μg LS), rePO (9.43 μg rePO), MntC (10 μg MntC), physical mixture (8.57 μg LS, 10 μg MntC, and 9.43 μg rePO), and MntC-rePO@LS (10.86 μg rePO-ST and 18.57 μg MntC@LS) groups, with 5 animals in each group. The mice from the above groups were immunized intramuscularly with corresponding agents dissolved in PBS buffer on days 0, 7, and 14. The vaccine was characterized by SDS-PAGE, size-exclusion chromatography, dynamic light scattering, and transmission electron microscopy (TEM). In vitro and in vivo safety of the vaccine was assessed through hemolysis assay, cytotoxicity test, body weight monitoring, histopathological examination, and hematological and serum biochemical analyses. The serum samples were collected on day 7 after the final immunization, and the titers and subtypes of anti-MntC and anti-rePO IgG were evaluated by indirect ELISA to evaluate the immunogenicity of the vaccine. Protective efficacy was assessed in mouse pneumonia models of single PA, single SA, and PA/SA co-infections by monitoring body weight and infection scores post-challenge, and by quantifying lung bacterial loads and observing histopathological changes at 24 h post-challenge.

We successfully constructed homogeneous MntC-rePO@LS nanoparticles with a hydrodynamic diameter of approximately 65.69 nm. The vaccine exhibited no significant hemolysis or cytotoxicity at concentrations ranging from 10 to 200 μg/mL. The immunized mice demonstrated normal body weight gain, with no pathological damage observed in major organs or at the injection site, and all hematological and biochemical parameters remained within normal ranges. Immunogenicity studies revealed that the MntC-rePO@LS group induced significantly higher anti-MntC and anti-rePO specific IgG titers at all 3 time points tested (day 0, 7, and 14) compared to monomer and physical mixture groups (P<0.05), with a predominant IgG2b subtype response. In the models of single PA, single SA, and co-infections, the MntC-rePO@LS group exhibited faster body weight recovery, lower infection scores, significantly reduced bacterial loads in the lungs (P<0.05), and markedly attenuated lung histopathological damage compared to all control groups.

The successfully constructed MntC-rePO@LS bivalent nanovaccine demonstrates favorable safety and immunogenicity, providing significant immune protection against single and co-infections with SA and PA. These findings validate its potential as a candidate vaccine to address antimicrobial resistance and polymicrobial infections.

Klebsiella pneumoniae (Kp) is a clinically prevalent opportunistic pathogen, with escalating antimicrobial resistance posing substantial challenges. Current vaccine strategies are constrained by the complexity of serotype diversity and the weak immunogenicity of candidate antigens, hindering effective protection. The construction of a multiepitope vaccine against K. pneumoniae, designated HBc-KpEpi, based on hepatitis B core antigen (HBc) virus-like particles, aims to overcome the weak immunogenicity of epitope peptides and to validate its ability to induce specific immune responses and protective efficacy.

After the multi-epitope vaccine HBc-KpEpi was expressed and purified, its purity and self-assembly status were characterized by SDS-PAGE, gel filtration chromatography, and transmission electron microscopy. Its safety was evaluated using cytotoxicity assay, hemolysis assay, and detection of serum biochemical indicators. Female BALB/c mice (6 to 8 weeks old, weighing 18 to 20 g) were randomly divided into the following groups: PBS control group, KpEpi group, and HBc-KpEpi group(n=10). Each group received intranasal immunization with PBS, KpEpi, or HBc-KpEpi (30 μg/mouse) on days 0, 14, and 21, respectively. In 7 d after the final immunization, the mice were challenged with a lethal dose (6×10⁶ CFU) or a sublethal dose (5×10⁵ CFU) of the clinical strain YBQ via intratracheal instillation. Detection indicators included survival rate and changes in body weight, bacterial colonization in lung tissue, serum inflammatory cytokines (IL-6, TNF-α, and IL-1β), lung histopathological changes, titers of serum specific antibodies (IgG, IgG1, IgG2a), and proportions of splenic CD4⁺ T cell subsets (Th1, Th2, Th17). Meanwhile, the ability of the vaccine to induce the differentiation and maturation of mouse bone marrow-derived dendritic cells (BMDCs) was evaluated through in vitro experiments.

The HBc-KpEpi protein was successfully prepared with high purity and self-assembled into regular particles. The survival rates in the HBc-KpEpi group were significantly higher than those in the PBS group (P<0.05). The bacterial colonization in lung tissue was significantly lower in the HBc-KpEpi group than that of the PBS and KpEpi groups (P<0.01). The HBc-KpEpi group exhibited significantly lower IL-6 level than the PBS group (P<0.000 1), with the levels of TNF-α and IL-1β showing downward trends. Lung histopathology showed that the alveolar structure remained relatively intact with markedly reduced inflammatory cell infiltration in the HBc-KpEpi group. After immunization, high titers of specific IgG antibodies were induced in the HBc-KpEpi group, with IgG1 as predominant subtype. Flow cytometry detection showed that the proportions of Th2 and Th17 cells in CD4⁺ T cells were significantly higher in the HBc-KpEpi group than in the KpEpi group (P<0.05) . In vitro experiments indicated that HBc-KpEpi significantly promoted the expression of MHC-Ⅱ, CD40, CD80, and CD86 on the surface of BMDCs (P<0.000 1)

The HBc-KpEpi vaccine significantly enhances the immunogenicity of epitope peptides, inducing cellular immune responses dominated by Th2 and Th17 as well as high titers of specific antibodies, and effectively reduce bacterial load, alleviate inflammatory damage, and improve the survival rate of mice.

Tissue-resident memory T (T_RM) cells play a key role as “immune sentinels” in mucosal immune protection. However, the molecular regulatory mechanisms underlying the formation and long-term residence of gastric CD4⁺ T_RM cells induced by Helicobacter pylori (H. pylori) vaccination remain unclear. This study aims to explore the key molecules and signaling pathways affecting T_RM formation, residence, and survival, with a particular focus on whether the Notch-RBPJ axis transcriptionally activates CD38, thereby revealing the specific molecular mechanisms underlying the formation and long-term maintenance of gastric CD4⁺ T_RM cells induced by H. pylori vaccination.

Single-cell RNA sequencing (scRNA-seq) was performed on gastric tissue CD4⁺ T cells following vaccination. Mice were vaccinated via subserosal injection in the gastric wall, with gastric tissue and splenic lymphocytes harvested on days 21 and 28 for multicolor flow cytometry analysis. Immunofluorescence co-staining was conducted on gastric tissue sections from mice on day 28 post-vaccination. H.pylori vaccine-immunized mice were continuously administered the RBPJ-specific inhibitor RIN1, and the changes in the proportions of CD4⁺ T cell subsets in gastric tissue were analyzed by flow cytometry. Primary murine CD4⁺ T cells were isolated via magnetic bead sorting and treatedwith the Notch pathway inhibitor RG-4733 or the RBPJ inhibitor RIN1 in an in vitro culture system. The treated primary CD4⁺ T cells were harvested for multicolor flow cytometry, and total RNA was extracted for qRT-PCR to assess the mRNA expression levels of target genes. The PGL3-CD38 promoter luciferase reporter plasmid and an RBPJ overexpression plasmid were constructed, with overexpression efficiency validated by qRT-PCR. The CD38 promoter reporter plasmids, RBPJ overexpression plasmids, and internal control plasmids were co-transfected into 293T cells, and luciferase activity was measured in 48 h later.

The Notch-RBPJ-CD38 axis was selected as the key factor influencing the establishment and residence of gastric epithelial CD4⁺ T_RM induced by H. pylori vaccine CCF/SF. On day 21 post-subserosal immunization with the H.pylori vaccine, gastric epithelial CD69⁺CD103⁺CD4⁺ T_RM cells exhibited significant activation of gene sets related to the Notch and Hedgehog signaling pathways and others compared to other gastric CD4⁺ T cell subsets, displaying the most pronounced differences. SCENIC analysis revealed a significant correlation between the core transcription factor RBPJ and CD38（P<0.05）. Beyond typical residency-associated molecules such as CD69 and CD103, elevated mRNA levels of Notch 1, RBPJ, Gzma, and Gzmb represented characteristic features distinguishing this subset from other CD4⁺ T cell populations. GO and pathway analyses suggested that CD4⁺ T_RM cells might possess dynamic proliferative potential. Subserosal surgical vaccination with the H. pylori vaccine CCF/SF upregulated the local expression of Notch signaling pathway-related protein Notch 1 and promoted protein expression of the key transcription factor RBPJ（P<0.05）. Blockade of the Notch pathway significantly reduced the proportion of resident T_RM cells in the stomach. In vitro experiments confirmed that Notch-RBPJ pathway inhibition decreased CD38 protein expression, thereby reducing TRM residency. RBPJ directly bound to the CD38 promoter to regulate CD38 expression（P<0.05）.

Following H. pylori vaccination, T_RM cells exhibit activation of the Notch signaling pathway, characterized by high expression of Notch 1 and RBPJ. This study confirms that the Notch-RBPJ signaling pathway transcriptionally regulates CD38 expression, representing a critical mechanism for the residency of gastric epithelial CD4⁺ T_RM cells.

To compare the differential activation characteristics of Toll-like receptor 5 (TLR5) agonist KFD and Toll-like receptor 7 (TLR7) agonist R837 on nasal epithelial cells (NECs) and bone marrow-derived dendritic cells (BMDCs), and to determine whether their combination produces synergistic enhancement as nasal mucosal adjuvants.

Mouse NECs and BMDCs were stimulated with 200 ng/mL KFD or R837 alone or in combination (n=3). At 3 h after stimulation, transcriptome sequencing was performed to analyze gene expression profiles, with key gene expression validated by qPCR. IL-6 levels in the supernatants were measured by ELISA at 12 h post-stimulation. Female C57BL/6 mice (6 to 8 weeks old, weighing 18 to 20 g) were randomly divided into negative control (normal saline), KFD alone (2 μg) group, R837 alone (2 μg) group, and KFD+R837 combination (2 μg KFD+2 μg R837) groups (n=5). At 18 h after intranasal administration, nasal lavage fluid (NALF) and bronchoalveolar lavage fluid (BALF) were collected to detect IL-6 levels, and the expression of costimulatory molecules CD80/CD86 on dendritic cells (DCs) in cervical lymph nodes (CLNs) was analyzed by flow cytometry. Using ovalbumin (OVA) as a model antigen for intranasal immunization, mice were randomly divided into normal saline (negative control), OVA alone (25 μg), OVA+KFD (25 μg OVA+2 μg KFD), OVA+R837 (25 μg OVA+2 μg R837), and OVA+combination adjuvant (25 μg OVA+1 μg KFD+1 μg R837) groups (n=5). Mice were immunized intranasally at weeks 0, 4, and 8. Serum and saliva samples were collected at 2 weeks after the final immunization, and OVA-specific IgG/IgA antibody levels were detected by ELISA.

Transcriptomic analysis revealed that KFD primarily activated NECs, with differentially expressed genes enriched in innate immune pathways such as TNF and NF-κB. In contrast, R837 potently activated BMDCs, with differentially expressed genes involved in TNF and IL-17 signaling pathways. In vitro BMDC experiments confirmed that R837 upregulated Tnf and Cxcl10 mRNA levels more strongly than KFD (P<0.01). The combination synergistically enhanced the expression of Tnf, Cxcl10, and Cd40 (P<0.001) and IL-6 secretion (KFD+R837 combination vs single agent, P<0.000 1). After intranasal administration, both KFD and R837 significantly upregulated CD80/CD86 expression on CLN DCs, with the combination group showing the most significant CD80 upregulation (KFD+R837 combination vs single agent, P<0.05). KFD alone significantly elevated IL-6 levels in NALF, which was not further enhanced by the combination. In the OVA immunization model, the combination adjuvant group exhibited significantly higher levels of serum OVA-specific IgG and salivary OVA-specific IgA than the OVA group (P<0.000 1), which were superior to either single adjuvant (serum IgG: OVA+ combined adjuvants vs OVA+R837, P<0.05; salivary IgA: OVA+ combined adjuvants vs OVA+KFD, OVA+ combined adjuvants vs OVA+R837, both P<0.05).

TLR5 and TLR7 agonists differentially activate NECs and DCs. Their combination synergistically activates DCs and significantly enhances both systemic and local antigen-specific immune responses to mucosal vaccines.

Intestinal organoids serve as important tools for drug screening, but the extent to which they recapitulate in vivo tissue microenvironments and drug responses remains inadequately evaluated. This study aims to verify systematic differences between intestinal organoids and in vivo intestinal crypts in fundamental biological characteristics and responses to nobiletin (NOB) through parallel comparison.

A standard mouse small intestinal organoid culture system was established, with synchronous isolation of homologous mouse small intestinal crypt tissues. Whole-transcriptome sequencing was employed to analyze gene expression profiles, flow cytometry was used to analyze cell subpopulation proportions, and Western blotting was performed to detect key protein expression levels. In vivo crypts and ex vivo organoids were treated with different concentrations of NOB: in vivo administration via gavage at 200 mg/kg for 12 h; organoids cultured for 48 h followed by 100 μmol/L NOB treatment for 12 h. GSEA, GO, and KEGG enrichment analyses and morphological observations were conducted on crypt cells from both groups to comprehensively evaluate the regulatory effects of NOB on cell cycle, metabolic flux, and signaling pathways.

Intestinal organoids effectively recapitulated core biological characteristics of in vivo crypts, including active proliferation of intestinal stem cells, multi-lineage differentiation potential, and CD24-marked cell clustering patterns. However, organoids exhibited specific metabolic reprogramming and immune-silent phenotypes, with differential glycosylation modification of the stem cell marker OLFM4. NOB treatment further amplified model-dependent differences: in in vivo crypts, NOB primarily downregulated the genes related to immune defense and oxidative stress, without significant effects on crypt morphology or OLFM4⁺ cell numbers; in organoids, NOB significantly suppressed core anabolic pathways including DNA replication, cell cycle, and amino acid and nucleotide synthesis, downregulated the expression of stem cell markers, and markedly reduced organoid budding number (P<0.001).

Intestinal organoids and in vivo tissues share commonalities while exhibiting differences at both baseline states and drug response levels. Organoids can recapitulate intestinal epithelial self-renewal processes but differ from in vivo tissues in metabolic programming and immune response. NOB mainly modulates immune microenvironment-related pathways in the in vivo context, whereas it directly targets epithelial cell-autonomous proliferation and metabolic programs in organoids.

Myelodysplastic syndromes (MDS) are associated with a high risk of progression to leukemia, highlighting the critical need to explore novel therapeutic agents. Based on network pharmacology prediction and molecular docking, we hypothesized that luteolin (LUT) might induce cellular senescence and apoptosis in MDS cells by targeting the Akt signaling pathway. This study aims to verify its targets and molecular mechanisms through experimental studies.

By integrating multiple databases, such as PharmMapper and GeneCards, common targets of LUT and MDS were screened. A protein-protein interaction (PPI) network was constructed using the STRING database and visualized with Cytoscape software to identify key targets. The DAVID database was employed for the functional enrichment analysis of Gene Ontology (GO) and the pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) for the common targets. Molecular docking between LUT and the key targets was performed using AutoDock software, and the docking efficacy was evaluated by analyzing the binding energy. Subsequently, molecular dynamics simulations were carried out using Gromacs to observe the ligand-receptor interactions and conformational changes. MUTZ-1 and MOLM-13 cells were used as experimental models. After treating the cells with different concentrations of LUT for 24 h （MUTZ-1：0、15、30、60 μmol/L，MOLM-13：0、4、8、16 μmol/L）, cell viability was measured by CCK-8 assay, DNA synthesis capacity was assessed by EdU assay, colony formation was examined by soft agar assay, morphological changes were observed by transmission electron microscopy (TEM), intracellular reactive oxygen species (ROS), mitochondrial ROS, mitochondrial membrane potential, and apoptosis rates were detected by flow cytometry, senescence was assessed by senescence-associated β-galactosidase (SA-β-Gal) staining, mRNA expression levels of senescence-associated secretory phenotype (SASP) factors were determined by RT-qPCR, and protein expression levels were examined by Western blotting.

A total of 568 LUT targets and 2 348 MDS-related targets were screened, yielding 228 intersecting targets. The intersecting targets were imported into the DAVID database. GO functional enrichment analysis demonstrated that biological processes were enriched in the responses to oxidative stress and cellular senescence; cellular components were enriched in protein kinase complex and secretory granule lumen; and molecular functions were enriched in protein kinase activity and cytokine receptor binding. KEGG pathway enrichment analysis revealed predominant enrichment in key pathways including the Akt signaling pathway, cellular senescence, and apoptosis. The PPI network constructed using the STRING database and Cytoscape software identified the top 10 key targets, as IL6, NFKB1, STAT3, TNF, JUN, BCL2, ALB, CASP3, Akt1 and IL1B. Molecular docking results indicated that the binding energy between LUT and Akt1 was -6.25 kcal/mol, suggesting a strong interaction. The molecular dynamics simulation results demonstrated stable binding between LUT and the Akt1 target. In vitro experimental results showed that LUT inhibited the proliferation of both MUTZ-1 and MOLM-13 cells in a dose-dependent manner. The half-maximal inhibitory concentration (IC₅₀) of LUT acting on MUTZ-1 cells for 24 h was 29.960 μmol/L, and that on MOLM-13 cells for 24 h was 8.654 μmol/L. Treatment of MUTZ-1 and MOLM-13 cells with various concentrations of LUT for 24 h significantly inhibited DNA synthesis and colony formation (P<0.01). The treatment of LUT at different concentrations for 24 h resulted in significant increases in intracellular ROS and mitochondrial ROS levels in a dose-dependent manner (P<0.05), decrease in mitochondrial membrane potential in a dose-dependent manner (P<0.05), and elevation of apoptosis rates in a dose-dependent manner (P<0.05). TEM revealed obvious apoptotic morphological features in MUTZ-1 and MOLM-13 cells after 24 h of LUT treatment. The protein levels of Cleaved PARP and Cleaved caspase-3, and the pro-apoptotic protein BAX were significantly upregulated, and that of the anti-apoptotic protein BCL-2 was significantly downregulated. SA-β-Gal staining showed that the proportion of senescent cells was significantly increased in MUTZ-1 and MOLM-13 cells after LUT treatment for 24 h, along with enhanced expression of related SASP factors (P<0.001), and significant upregulation of senescence-associated proteins p53, p21 and p16. After LUT treatment for 24 h, the protein expression of p-Akt (Ser473) and p-MDM2 (Ser166) was significantly downregulated, while the total Akt protein expression showed no significant change.

LUT may induce mitochondria-dependent apoptosis and senescence in MDS cells by targeting the Akt signaling pathway, thereby inhibiting the proliferation of MDS cells.

Posterior communicating artery aneurysm(PCoAA) exhibit complex anatomical relationships, and microsurgical clipping of large or posteromedially projecting aneurysms remains technically challenging with considerable postoperative morbidity. Therefore, selecting optimal surgical approach has become a key clinical concern. This study compares the efficacy, safety, and overall performance of the Dolenc approach versus the conventional pterional approach for clipping large or posteromedially projecting PCoA aneurysms.

A prospective nonrandomized controlled study design was adopted. A total of 79 patients with large or posteromedially projecting PCoA aneurysms consecutively treated in Department of Neurosurgery, Liangping Hospital of the First Affiliated Hospital of Chongqing Medical University between September 2019 and September 2025 were enrolled, and alternatively assigned to the Dolenc approach group (n=41) or conventional pterional approach group (n=38) based on admission sequence. The primary endpoint was the favorable outcome rate at 3 months postoperatively, assessed using Glasgow Outcome Scale (GOS), with GOS score of 4 to 5 defined as a favorable outcome and score of 1 to 3 as an unfavorable outcome. Secondary endpoints included operative time, postoperative length of hospital stay, distribution of GOS scores, and aneurysm neck residual on digital subtraction angiography (DSA)/CT angiography (CTA) at 3 months postoperatively (defined as contrast filling of the aneurysm neck). Firth’s bias-reduced penalized likelihood logistic regression analysis was used to evaluate the association between surgical approach and unfavorable outcome (with age stratification as covariate), reporting odds ratio (OR), 95% confidence interval (CI), and P value. In addition, a standardized three-domain (technical-safety-efficiency) scoring system and a composite score were constructed for descriptive comparison, along with cost-effectiveness and subgroup analyses.

For the primary endpoint, the favorable outcome rate was 97.6% (40/41) in the Dolenc group and 78.9% (30/38) in the conventional group, with an absolute difference of 18.6% (95%CI: 4.8% to 32.4%, P=0.009). Postoperative length of hospital stay was shorter in the Dolenc group (13.46±2.43 vs 16.05±2.46 d), with a mean difference of -2.59 d (95%CI: -3.69 to -1.49, P<0.001). Operative time was comparable between groups (3.98±0.60 vs 3.97±0.62 h), with a mean difference of 0.01 h (95%CI: -0.26 to -0.28, P=0.944). Regarding functional outcomes, the Dolenc group had a greater proportion of patients achieving a GOS score of 5 than the conventional group (61.0% vs 18.4%), with an absolute difference of 42.6% (95%CI: 23.2% to 61.9%, P<0.001), and the distribution of GOS scores in the Dolenc group was shifted toward better outcomes, with lower proportions of patients having GOS scores of 4 and 3 than the conventional pterional group (χ²=14.96, P<0.001). For imaging follow-up, the neck remnant rate was lower in the Dolenc group (2.4% vs 13.2%), with an absolute difference of -10.8% (95%CI: -22.5% to -1.0%, P=0.072). Multivariable analysis showed that the conventional pterional approach was associated with unfavorable outcome (OR=10.20, 95%CI: 1.20 to 87.04, P=0.034). The model was significant overall (χ²=7.56, df=2, P=0.02), with good calibration (Hosmer-Lemeshow test P=0.82), and moderate discrimination (AUC=0.75, 95%CI: 0.61 to 0.86). Both the three-domain scores and the composite score were higher in the Dolenc group (composite score: 0.873 vs 0.739). Subgroup analyses suggested the Dolenc approach demonstrated greater advantage in high-risk/complex subgroups, with difference of 0.256 for Hunt-Hess grade Ⅲ-Ⅳ (95%CI: 0.056 to 0.475, P=0.025) and 0.157 for large aneurysms (95%CI: 0.016 to 0.305, P=0.049); however, these findings should be interpreted with caution given the limited sample sizes in some subgroups.

In this cohort, for microsurgical clipping of large or posteromedially projecting PCoA aneurysms, the Dolenc approach is associated with a higher rate of 3-month favorable postoperative outcomes and a shorter length of hospital stay, without an apparent increase in operative time.

“One-stop” interventional therapy for non-valvular atrial fibrillation (NVAF) traditionally relies on transesophageal echocardiography (TEE) guidance, which carries limitations including anesthesia requirements and esophageal injury risks. Whether digital subtraction angiography (DSA) combined with transthoracic echocardiography (TTE) guidance can balance therapeutic efficacy with procedural simplification remains unclear. This study aims to compare the safety and efficacy of “one-stop” procedure for NVAF under different imaging guidance modalities.

Based on predefined inclusion and exclusion criteria, this retrospective cohort study was conducted on consecutive NVAF patients undergoing combined “one-stop” interventional therapy in Department of Cardiology of our hospital between August 1, 2014 and July 31, 2024. According to intraoperative guidance methods, they were divided into the DSA combined with TTE group (DSA group, n=77) and the DSA combined with TEE group (TEE group, n=209). Relevant clinical data of the patients were collected from the hospital big data intelligent platform medical record system, including gender, age, comorbidities, and results of TTE and TEE examinations. Postoperative follow-up included atrial fibrillation recurrence, stroke, bleeding, device-related thrombosis (DRT), cardiac tamponade, hospitalization due to heart failure, and cardiac death. The safety and efficacy of the “one-stop” procedure were compared between the 2 groups.

There were 286 patients enrolled in this study, including 170 males and 116 females, with 77 in the DSA group and 209 in the TEE group. All patients completed at least 6 months of follow-up, with a mean duration of 36.49±14.26 months. No statistically significant differences were observed in baseline characteristics between the 2 groups (P>0.05). The DSA group demonstrated shorter mean procedural time (202.4±39.3 vs 214.1±33.4 min, P=0.013). The incidence of DRT was higher in the DSA group than in the TEE group (3.9% vs 1.4%), though the difference was not statistically significant (P=0.411). Atrial fibrillation recurrence occurred in 10 patients (13.0%) in the DSA group and 19 patients (9.1%) in the TEE group, with no significant intergroup difference. Regarding clinical endpoints, no significant differences were observed between the 2 groups in terms of stroke, major bleeding, cardiac tamponade, heart failure rehospitalization, or cardiac death. Kaplan-Meier survival curve analysis revealed no statistical differences between the 2 groups in composite endpoints, such as major adverse cardiovascular events (MACE), and in the efficacy composite endpoint, including atrial fibrillation recurrence, heart failure rehospitalization, stroke, or transient ischemic attack.

For NVAF patients undergoing one-stop procedures, DSA guidance can shorten procedural time and achieve comparable safety and efficacy to TEE guidance, with the exception of a trend toward increased DRT incidence.

Infertility represents a major global reproductive health challenge. Metabolite levels in both partners are highly correlated and may serve as potential confounders for each other. However, how the overall preconception plasma metabolic profiles of both partners jointly influence natural conception outcomes remains unclear. Based on the Chongqing Preconception Reproductive Health and Birth Outcomes Cohort (PREBIC), this study aims to explore the association between preconception plasma metabolite levels in couples of reproductive age and conception outcomes.

The Chongqing PREBIC was launched in January 2019 at the Chongqing Health Center for Women and Children, recruiting couples of reproductive age undergoing preconception health examinations to establish baseline cohorts and collect blood samples. Follow-up was then performed every 3 months for clinical pregnancy outcomes following 24 months of unprotected intercourse. The study ultimately included 1 292 couples of reproductive age for analysis. According to natural conception status, participants were assigned into a pregnant group (n=967) and a non-pregnant group (n=325). Baseline plasma samples were subjected to standardized metabolomic testing using the Bruker IVDR NMR platform, yielding 167 metabolic parameters. Elastic network regression was employed to screen key metabolites, and logistic regression models were constructed to evaluate the association between metabolites and 2-year pregnancy outcomes, and confounding factors such as age, body mass index (BMI), smoking and alcohol consumption were adjusted.

Among the 1 292 couples of reproductive age, the 2-year pregnancy rate was 74.85%. Eighty-two metabolites showed positive correlations between partners, with a mean correlation coefficient of 0.15. After controlling for inter-variable correlations using elastic network model, 14 male metabolites and 20 female metabolites were found to be associated with conception outcomes. Logistic regression revealed gender-specific associations between plasma metabolite levels in both partners and conception outcomes: in males, triglycerides in low-density lipoprotein were positively associated with conception outcomes (Q4 vs Q1: RR=1.12, 95%CI: 1.03 to 1.19, P adjust=0.047). In females, low-density lipoprotein, apolipoprotein A in high-density lipoprotein, apolipoprotein B in low-density lipoprotein, creatinine, and leucine were negatively associated with conception outcomes. Acetic acid in in both partners was positively correlated with conception outcomes (Q4 vs Q1: male RR=1.13, 95%CI: 1.04 to 1.21, P adjust=0.047; female RR=1.18, 95%CI: 1.05 to 1.28, P adjust=0.007).

Preconception metabolite levels in couples of reproductive age may act on future reproductive potential in a gender-specific manner, with common key metabolites identified. Countermeasures　Based on these findings, it is recommended that the health policy-making departments accelerate the transition of preconception care services from focusing solely on women toward a dual-partner co-management model. Incorporate the pre-pregnancy metabolism of both males and females of reproductive age into routine preconception eugenics health services, with priority monitoring of metabolic indicators, including lipid homeostasis and energy metabolism; develop precision intervention measures according to metabolic differences between genders; improve the urban-rural integrated preconception health management system covering both partners; and enhance reproductive health at the public health source level, providing policy basis and technical support for optimizing birth population quality.

China has achieved remarkable progress in tuberculosis (TB) control, with the incidence rates declining continuously; however, the disease burden among females remained substantial with pronounced regional heterogeneity. The differential impacts of climatic factors on TB onset and their driving mechanisms remains poorly understood, particularly lacking zone-specific evidence based on climatic gradients. This study aims to analyze the heterogeneous effects of climatic factors and air pollutants on female TB incidence across 6 climatic zones and identify key driving pathways, thereby providing evidence-based support for developing gender-sensitive, climate-adapted precision prevention and control strategies.

A retrospective descriptive study design was adopted. Based on China’s climate zoning standards and typical climate characteristics, 6 representative regions were selected, corresponding to the 6 major climate zones: Cold-Dry Zone, Hot-Humid Zone, Warm-Humid Zone, Temperate Continental Zone, Alpine Zone, and Temperate Monsoon Zone. Sample representativeness was evaluated across 3 dimensions: geographic coverage (encompassing northern, southern, eastern, western, and central China), climate consistency (consistent with the core climate characteristics of the corresponding regions), and population epidemiological representativeness. A total of 9 570 female patients with newly diagnosed TB in 6 regions from 2017 to 2024, along with official meteorological and pollutant monitoring data during the same period, were collected. Two categories of measurement indicators were established for cases and meteorological pollution variables. After preprocessing such as imputation of missing values and construction of lagged 1 to 3 period variables, a panel data set was formed. With female TB case counts as the dependent variable, stepwise regression analysis was applied, and model performance was evaluated using R², VIF, and F/t tests.

The overall demographic and clinical characteristics of female TB patients exhibited highly significant heterogeneity across the 6 climatic zones (P<0.001). The TB incidence in the females increased with advancing age, showing a fluctuating downward trend across all regions with significant inter-zone differences. The alpine zone demonstrated high incidence rates, initially declining then rising, decreasing from 155/100 000 in 2017 to 36/100 000 in 2022, subsequently increasing to 91/100 000 in 2024. Regression models for all climatic zones passed multicollinearity tests (VIF<10) and demonstrated extremely significant statistical significance (P<0.001): R²=0.452 for the Cold-Dry Zone, R²=0.589 for the Hot-Humid Zone, R²=0.610 for the Warm-Humid Zone, R²=0.626 for the Temperate Continental Zone, R²=0.641 for the Alpine Zone, and R²=0.580 for the Temperate Monsoon Zone. The combined model yielded R²=0.584 (F=71.548, P<0.001). Lag effects of 2 to 3 periods were present across all regions. Core driving factors were zone-specific, with lag 1 period PM10 identified as the core positive driver of female TB onset.

Climatic conditions and air pollutants exert significantly heterogeneous effects on female TB incidence across China’s 6 climatic zones. The 2 to 3 period lag effects essentially align with the incubation period of Mycobacterium tuberculosis infection. Regional differences in the incidence are closely associated with climatic characteristics, population behavioral patterns, and pollutant exposure profiles. Countermeasures Building upon the existing technical guidance system for TB prevention and control, it is recommended to incorporate early warning rules tailored to local climatic conditions and pollutant lag-based screening window standards. Screening resources and health education efforts at the primary care level should be reallocated based on core pollutant drivers in each region. Data interfaces among meteorological, environmental, and TB surveillance systems should be integrated to establish linked assessment mechanisms, enabling zone-specific, time-specific precision early warning and targeted interventions.

The pattern of socioeconomic status (SES) inequality across the natural history continuum of cervical cancer (from precancerous lesions to invasive cancer) remains unclear. This study aims to systematically investigate the associations between different dimensions of SES and risk at various stages of cervical lesions, providing scientific evidence for identifying high-risk populations and formulating targeted prevention and control strategies.

A case-control design was adopted. A total of 508 female patients who visited Department of Gynecology at the First Affiliated Hospital of Army Medical University between October 2024 and September 2025 were enrolled. Based on pathological diagnosis combined with international classification of diseases， 10th revision（ICD-10) coding, cervical intraepithelial neoplasia (CIN), cervical carcinoma in situ, and invasive cervical cancer were designated as 3 case groups, with the women free of cervical lesions serving as the control group. Four SES indicators, that is, education level, household income, occupational income, and area-level economic development, were collected via questionnaires. Given that the outcome, variable was ordinal and the proportionul odds assumption was violated three independent binary logistic regression models were constructed to analyze the associations between each SES indicator and different lesion stages, estimating ORs and 95%CIs, with adjustment for age, body mass index (BMI), and human papillomavirus (HPV) vaccination status as confounders. Multicollinearity was assessed using variance inflation factor (VIF), and sensitivity analysis was performed using multinomial logistic regression.

A total of 508 patients were enrolled, including 74 (14.6%) in the lesion-free group, 213 (41.9%) in the CIN group, 110 (21.7%) in the carcinoma in situ group, and 111 (21.8%) in the invasive cervical cancer group. Multivariate logistic regression analysis revealed that high school education and above was associated with reduced risks of CIN (OR=0.475, 95%CI: 0.258 to 0.877, P=0.017) and invasive cervical cancer (OR=0.361, 95%CI: 0.175 to 0.746, P=0.006), but showed no statistically significant association with carcinoma in situ. occupational income >5 000 yuan was associated with a reduced risk of invasive cervical cancer (OR=0.332, 95%CI: 0.137 to 0.806, P=0.015). Women from non-affluent areas exhibited a 92% increased risk of invasive cervical cancer compared with those from affluent areas (OR=1.920, 95%CI: 1.005 to 3.668, P=0.048). No significant associations were observed between household income and cervical lesions at any stage. All VIF values for variables in Model 3 were <1.12, indicating absence of multicollinearity. Sensitivity analysis using multinomial logistic regression with the lesion-free group as the common reference showed that the relative risk ratios (RRRs) of SES indicators across lesion stages were consistent in direction with the independent binary models, suggesting robust findings.

The impact of socioeconomic disparities on cervical lesion risk among Chinese women exhibits dimensional heterogeneity. High school education and above is independently associated with reduced risks of CIN and invasive cervical carcinoma, whereas residence economic level and occupational income primarily influence the risk of invasive cervical cancer. Countermeasures Integration of screening into grassroots family planning systems, implementation of data-driven bottleneck analysis and micro-interventions, and strengthening of community mobilization and family engagement are recommended to systematically eliminate accessibility barriers faced by populations with low education level and in non-affluent areas regarding HPV vaccination, screening, and treatment, ensuring equitable progress toward cervical cancer elimination goals for disadvantaged populations.