ObjectiveTo explore the mechanism of Yishen Qubi Tongluo Formula （益肾祛痹通络方， YQTF） in the treatment of rheumatoid arthritis（RA）. MethodsNetwork pharmacology was employed to retrieve and screen the active components and potential targets of YQTF as well as RA-related targets using databases including TCMSP， BATMAN， ETCM and GEO. The intersection of targets related to active components and RA-related targets was identified， and a protein-protein interaction （PPI） network was constructed. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed， and a drug-active component-common target network of YQTF in the treatment of RA was established. The core components of YQTF were molecularly docked with key targets. Human rheumatoid arthritis synovial fibroblast cell line MH7A was divided into blank group， model group， methotrexate group and YQTF group. The blank group was cultured with 10% fetal bovine serum， while the other three groups were stimulated with 10 μg/L of recombinant human tumor necrosis factor-α （TNF-α） for 24 h to establish the RA cell model. On this basis， the methotrexate group was treated with methotrexate suspension at a concentration of 20 μmol/L， and the YQTF group was treated with 10% YQTF-medicated serum. After 48 h of intervention， the levels of TNF-α and interleukin-17A（IL-17A）contents in cell supernatants were detected by enzyme-linked immunosorbent assay （ELISA）， and mRNA expressions of phosphatidylinositol 3-kinase（PI3K）， protein kinase B（AKT） and mammalian target of rapamycin（mTOR） were detected by real-time quantitative polymerase chain reaction （RT-qPCR）. ResultsNetwork pharmacological analysis identified 209 active components and 583 potential target genes of YQTF， as well as 818 RA-related targets. A total of 29 common targets were obtained from the intersection of drug-related targets and RA-related targets. Quercetin，β-sitosterol， kaempferol， stigmasterol and luteolin were the core active components of YQTF for the treatment of RA， while matrix metalloproteinase-9 （MMP9）， prostaglandin-endoperoxide synthase 2 （PTGS2）， Toll-like receptor 4 （TLR4）， tumor protein p53 （TP53） and transcription factor AP-1 subunit JUN were the key targets. The GO and KEGG pathway enrichment analysis showed that the involved biological processes and pathways were mainly associated with antioxidant responses， PI3K-AKT signaling pathway and Toll-like receptor （TLR） signaling pathway. Molecular docking results showed that MMP9 and PTGS2 exhibited high binding affinities with quercetin， β-sitosterol， kaempferol， stigmasterol and luteolin； TLR4 exhibited high binding activities with β-sitosterol， stigmasterol and luteolin； and TP53 showed high binding affinity with luteolin. The results of cell experiments showed that compared with the control group， the contents of TNF-α and IL-17A as well as the mRNA expressions of AKT and mTOR in the model group significantly increased （P＜0.05 or P＜0.01）. Compared with the model group， all the above indicators significantly decreased in the YQTF group， while the contents of TNF-α and the mRNA expression of AKT significantly decreased in the methotrexate group （P＜0.05 or P＜0.01）. ConclusionThe mechanism of YQTF in the treatment of RA may be associated with reducing inflammatory cytokine secretion and inhibiting the activation of the PI3K-AKT-mTOR signaling pathway.

Asthma and cough variant asthma （CVA） are both chronic heterogeneous diseases characterized by airway microenvironment homeostasis disruption as their core pathological basis. In recent years， micro ribonucleic acid （miRNA）， as core post-transcriptional regulators， have been shown to finely modulate multiple critical signaling pathways， including Janus kinase/signal transducer and activator of transcription （JAK/STAT）， nuclear factor-κB （NF-κB）， transforming growth factor-β/Smad （TGF-β/Smad）， and phosphoinositide 3-kinase/protein kinase B （PI3K/Akt）， as well as various pathological processes such as airway epithelial barrier restoration， type 1 helper T cell（Th1）/Th2 immune balance， M1/M2 macrophage polarization， airway smooth muscle cell function， and airway hyperresponsiveness. miRNAs play a pivotal role in maintaining and disrupting airway microenvironment homeostasis. Based on recent Chinese and international literature， a logical framework centered on "airway microenvironment homeostasis disruption， miRNA regulation， and microenvironment restoration" was constructed. From the perspective of the airway microenvironment， the therapeutic roles of miRNA in asthma and CVA were systematically summarized， and the cascade regulatory mechanisms of miRNA throughout the entire disease course were elucidated. The hub miRNA was identified， and research progress on traditional Chinese medicine intervention strategies was explored. Furthermore， current clinical studies on RNA therapeutics and traditional Chinese medicine in achieving multi-target and multi-pathway integrated treatment by modulating miRNA were analyzed. The value of miRNA as biomarkers for diagnosis， phenotyping， and prognosis assessment， as well as the potential and application prospects of miRNA mimics and antagonists in precision therapy， were summarized， with the ultimate goal of advancing precision therapy for asthma and CVA.

Objective:To analyze the contamination characteristics and pathogenic potential of Listeria monocytogenes (LM) in pre-packaged and refrigerated ready-to-eat cooked meat products in Chengdu City. Methods:From September 2022 to May 2023, pre-packaged and refrigerated ready-to-eat cooked meat products were collected from six districts and counties in Chengdu City. Qualitative and quantitative determination of LM was performed on these samples. Whole genome sequencing was carried out on the isolated strains. Different ST strains were selected for cell adhesion and invasion experiments. The results were expressed as adhesion rate and invasion rate. One-way ANOVA was used for comparison between groups, and the Dunnett t-test was used for pairwise comparison. Results:A total of 145 samples were collected, and LM was detected and isolated in 29 samples. The total detection rate was 20.00%, and the detection rate of braised pork was the highest (68.18%). The contamination level of LM in 9 samples (31.03%) was greater than 100.00 MPN/g. The 29 strains of Listeria monocytogenes belonged to 7 STs, including ST 3 (27.59%), ST 8 (17.24%) and ST 87 (13.79%). The strain of ST 87 carried Listeria pathogenicity islands 4 (LIPI-4), which was a highly virulent strain. The medium and high virulence strains accounted for 79.31%. Some moderately virulent and highly virulent strains of ST 8 and ST 87 were closely related to clinical patient strains. Some LM isolates of the same ST type had little SNP differences (1-6) in the same manufacturer at different stages. In vitro cell experiments showed that the highly virulent strain ST 87 had the strongest adhesion and invasion ability towards Caco-2 cells. Conclusion:The pre-packaged and refrigerated cooked meat products have a high contamination rate. Some samples have high contamination levels and carry medium and high virulence strains, posing potential health risks to human beings. LM residues continue to persist in some manufacturers′ production and processing stages.

Randomized controlled trials have long been regarded as the gold standard for evaluating vaccine efficacy. However, conducting randomized controlled trials to assess vaccines in real-world settings is often impractical due to limitations in research resources and ethical considerations. In recent years, the emulated target trial has emerged as a novel methodological framework that applies the design principles of randomized trials to observational studies. By using observational data to emulate the randomization and treatment assignment of a clinical trial, this approach enables the evaluation of vaccine effectiveness in the real world. The application of emulated target trials enhances the quality and applicability of real-world evidence. This article explains the fundamental principles, design elements, advantages, and limitations of using emulated target trials to evaluate vaccine effectiveness, providing methodological guidance for conducting such studies in China.

Objective:To investigate, for multi-sensitized allergic rhinitis (AR) patients allergic to dust mites combined with other allergens (pollen, mold, animal dander, etc.), whether the single dust mite subcutaneous immunotherapy (SCIT) can improve the specific symptoms caused by other allergens in the patients, and to analyze the relationship between the effectiveness of symptom improvement in these patients and the type, quantity and severity of the allergens.Methods:A multicenter retrospective study was conducted to collect mul-sensitized AR patients from allergy or respiratory departments of 5 hospitals who received house dust mite allergen preparation SCIT for 12 to 36 months and met other inclusion and exclusion criteria from February to July 2024. General clinical data were collected and the perennial or seasonal symptoms before and after treatment were evaluated with visual analogue scale (VAS) to assess whether there was an perennial or allergen-specific symptom improvement (VAS score decrease ≥30%), by which the patients were divided into effective group and ineffective. R software was used to analyze the differences between groups by using Fisher′s exact test and Mann-Whitney U test. Results:A total of 62 patients were enrolled, and the treatment were effective in 39 of them, with an effective rate of 62.9%. For allergen-specific symptoms, the median age of the effective group was higher than that of the ineffective group (12 years old vs. 8 years old, P=0.039), and the effective rate in dust mite specific immunoglobin E (sIgE) grade ≤5 group was higher than that in sIgE grade >5 group (81.6% vs. 45.5%, P=0.008), and the effective rate of mold sIgE grade ≤2 group was higher than that of sIgE grade >2 group (83.3% vs. 28.6%, P=0.045), and there was no statistically significant correlation between the other allergen grades and the effective rate ( P>0.05). For perennial symptoms, the effective rate in the mold grade ≤2 group was higher than that in the sIgE grade >2 group (91.3% vs. 28.6%, P=0.010), and there was no statistically significant correlation between the other allergen grades and the effective rate ( P>0.05). There was no significant correlation between the treatment effectiveness of perennial or allergen-specific symptoms and the number of combined allergens, the grade of skin test, and the difference between the grade of combined allergens and that of dust mites ( P>0.05). Conclusion:Among the patients with multi-sensitized AR allergic to dust mites included in this study, single dust mite SCIT is effective in some of them, and for allergen-specific symptoms, the effective group was elder, and dust mite sIgE grade 6 and mold sIgE grade ≥2 was related to the low effective rate of SCIT. The present results are insufficient for selecting single or multiple AIT in any type of multi-sensitized patients.

Objective To explore the mechanism by which She-Ti-Zhi-Qiu decoction alleviates allergic rhinitis (AR) through gut microbiota-mediated regulation of T helper cell 17(Th17)/regulatory T cells(Treg) balance and related cytokines. Methods Twenty-eight female SD rats were randomly divided into four groups: the Control group, Model group, STZQ group, and Probiotics group. Except for the Control group, all other groups were sensitized with ovalbumin (OVA) to establish AR models. The Control and Model groups received intragastric administration of normal saline, while the STZQ group was administered She-Ti-Zhi-Qiu Decoction, and Probiotics group received probiotics. After two weeks of continuous intragastric administration, nasal mucosa, serum, peripheral blood, and colon contents were collected. The inflammation of nasal mucosal tissue was assessed via HE staining. 16S rDNA sequencing was used to detect and analyze the structure and content of bacteria in colon contents. Flow cytometry was used to detect the relative proportions of Treg and Th17 cells in peripheral blood. ELISA was used to measure the levels of Th17- and Treg-related cytokines in serum. Results Compared with the Control group, the Model group showed an inflammatory response in nasal mucosal tissue, along with increased IL-17A and IL-17E levels and decreased IL-10 levels. The percentage of Th17 cells in peripheral blood increased, while the percentage of Treg cells decreased. Beneficial bacteria in the intestine were decreased, while pathogenic bacteria were increased. Compared with the Model group, the STZQ group showed lower serum IL-17A and IL-17E levels and higher IL-10 levels. The percentage of Th17 in peripheral blood decreased, while the percentage of Treg increased. There was an increase in beneficial bacteria in the intestine and a decrease in pathogenic bacteria. The changes in the microbiota were correlated with IL-17A, IL-17E, and IL-10 levels. Conclusion She-Ti-Zhi-Qiu decoction can ameliorate the inflammation of AR by regulating gut microbiota and Th17/Treg immune balance.
Animals ; T-Lymphocytes, Regulatory/drug effects* ; Th17 Cells/drug effects* ; Gastrointestinal Microbiome/drug effects* ; Rats, Sprague-Dawley ; Drugs, Chinese Herbal/therapeutic use* ; Female ; Rhinitis, Allergic/microbiology* ; Rats ; Cytokines

Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Objective To investigate the feasibility of parallel capillary bundle arrays for physiomimetic impedance modeling and establish a parametric quantification framework,thereby providing a customizable impedance characterization methodology for diverse in-vitro mock circulation researches.Methods Based on the parallel flow resistance and Poiseuille equation,a tube resistance element with multiple parallel-aligned capillary glass tubes was designed and fabricated.The resistance values of the capillary-bundle and a ball valve were measured through constant flow experiments analogous to electrical resistance measurement method.Moreover,a simple lumped-parameter mock circulation loop was constructed and the pressure and flow rate for each node of the loop were measured under different input flow waveforms.An 0D-Windkessel model corresponding to the experiment was developed.The impedance and compliance were adjusted to match the simulated and experimental pressure and flow waveforms.The accuracy of the capillary bundle impedance in pulsatile experiments was verified by using the computational resistance values.Results The constant-flow impedance calibration experiments revealed that the capillary bundle impedance remained unaffected by flow rate variations over a wide flow range.When the capillary bundle impedance was integrated into the pulsatile circulatory system and the same impedance value obtained from the constant-flow calibration was applied in the computational model,the resulting pressure and flow waveforms showed good agreement with those measured in the pulsatile experiments.However,when the ball valves with nominally identical impedance values were inserted in the pulsatile system,the calculated impedance exhibited a two-fold difference,and significant discrepancies were observed between the simulated and experimental terminal flow waveforms.Conclusions The capillary bundle impedance maintains a constant value regardless of flow rate variations.Once the calibrated resistance value is determined through constant flow experiments,it can be directly applied to pulsatile systems.This approach can provide quantitative pulsatile flow conditions for testing various medical devices.

Objective To conduct a quantitative risk assessment of Listeria monocytogenes(LM)in prepackaged,non-vacuum sealed,short shelf-life,ready-to-eat meat products in Chengdu using the@Risk software.Methods Based on monitoring data of LM contamination in pre-packaged,non-vacuum sealed,refrigerated ready-to-eat meat products in Chengdu obtained from a previous study,a risk assessment model was established.The risk of LM infection caused by consuming cooked meat products in different groups of people was quantitatively assessed.In addition,the growth of LM in cooked meat products,from retail to consumption,was also taken into consideration in the assessment.Results In Chengdu,the numbers of potential cases of listeriosis caused by consumption of prepackaged,non-vacuum sealed,refrigerated ready-to-eat meat products were 0.01(95％confidence interval[CI]:0-1.71 × 10-2)per year per million in healthy individuals aged 5-＜65 years old,0.22(95％CI:0-2.67 × 10-1)in healthy individuals aged 65 and above,and 2.88(95％CI:3.85 × 10-8-4.35)in pregnant women.According to the results of the sensitivity analysis,the initial pollution level of LM in the retail stage was the most important factor affecting the prevalence(R=0.25),followed by retail temperature(R=0.08),retail time(R=0.07),and amount of consumption per meal(R=0.07).Conclusions For pre-packaged,non-vacuum sealed,cooked meat products,the most important measure to reduce the prevalence of listeriosis is to control the initial contamination level,which requires food processing plants to regularly clean and strictly disinfect the processing environment and equipment to minimize LM contamination at the source.Retail delicatessens should strictly maintain a storage temperature below 5.0℃ and strictly adhere to product shelf-life recommendations.As for consumers,they should consume these meat products as soon as possible after purchase or store them under refrigerated conditions and shorten the storage time.Pregnant women should thoroughly heat the meat products before eating to reduce the risk of listeriosis.