ObjectiveTo observe the effects of Yiqi Huoxue Jiedu formula（YHJF） on immune cell exhaustion in the spleen of septic mice and to explore and validate its potential intervention targets. MethodsMice were randomly divided into the sham-operated， model， low-dose YHJF（4.1 g·kg^-1）， and high-dose YHJF（8.2 g·kg^-1） groups. Except for the sham-operated group， a cecal ligation and puncture（CLP） procedure was performed to establish a mouse sepsis model. The treatment groups received oral administration of the corresponding doses， while the sham-operated and model groups received an equal volume of physiological saline. After the intervention， the 7-day survival rate of each group was recorded， and spleen samples were collected 72 h post-intervention， and the spleen index was calculated. Terminal deoxynucleotidyl transferase deoxyuridine triphosphate（dUTP） nick end labeling（TUNEL） staining was used to detect apoptosis in spleen cells. Enzyme-linked immunosorbent assay（ELISA） was performed to measure the levels of interleukin（IL）-4 and IL-10 in the serum. Transcriptomics and metabolomics were used to screen for differentially expressed genes（DEGs） and differential metabolites in the spleen， followed by bioinformatics analysis to identify key targets. Real-time quantitative polymerase chain reaction（Real-time PCR）， flow cytometry， and multiplex immunofluorescence were used to verify the expressions of key genes and proteins. ResultsThe high-dose YHJF group significantly improved the 7-day survival rate of septic mice（P0.05）. Compared with the sham-operated group， the model group showed a significant increase in apoptosis of spleen cells and a decrease in the spleen index at 72 h post-modeling， with markedly elevated peripheral serum IL-4 and IL-10 levels（P0.01）. Compared with the model group， the high-dose YHJF group showed a reduction in apoptosis of spleen cells， an increase in the spleen index， and a significant decrease in peripheral serum IL-4 and IL-10 levels（P0.05）. Spleen transcriptomics identified 255 DEGs between groups， potentially serving as intervention targets for YHJF. Gene Ontology（GO） enrichment analysis revealed that DEGs were mainly involved in biological processes such as natural killer（NK） cell-mediated positive immune regulation， cell killing， cytokine production， positive regulation of innate immune cells， and interferon production. Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis showed that DEGs were mainly involved in cytokine-cytokine receptor interactions， viral protein interactions with cytokines and cytokine receptors， chemokine signaling pathway， and nuclear transcription factor-κB（NF-κB） signaling pathway. Protein-protein interaction（PPI） network analysis identified CD160， granzyme B（GZMB）， and chemokine ligand 4（CCL4） as key targets for YHJF in treating sepsis. Metabolomics identified 46 differential metabolites that were significantly reversed by YHJF intervention， and combined transcriptomics and metabolomics analysis identified 17 differential metabolites closely related to CD160. Pathway enrichment revealed that these metabolites were mainly involved in glycerophospholipid metabolism， arachidonic acid metabolism， glycosylphosphatidylinositol（GPI） anchor biosynthesis， linoleic acid metabolism， and α-linolenic acid metabolism pathways. Verification results showed that， compared with the sham-operated group， the model group exhibited significantly elevated CD160 mRNA expression level in the spleen， along with markedly decreased CCL4 and GZMB mRNA expression， and had a significant increase in CD160 expression on the surface of natural killer T（NKT） cells in the spleen（P0.01）. Compared with the model group， the high-dose YHJF group had a significant decrease in CD160 mRNA expression in the spleen， a significant increase in CCL4 and GZMB mRNA expressions. Further flow cytometry and immunofluorescence revealed that compared with the sham-operated group， CD160 expression on the surface of splenic NKT cells in the model group was significantly increased（P0.01）， while high-dose YHJF intervention significantly reduced CD160 expression（P0.01）. ConclusionYHJF may alleviate NKT cell exhaustion in sepsis by downregulating the expression of the negative co-stimulatory molecule CD160， and this regulatory effect is closely related to fatty acid metabolism pathways. This study provides new insights and targets for further exploration of strengthening vital Qi and detoxifying strategy to improve immune cell exhaustion in acute deficiency syndrome of sepsis.

ObjectiveTo observe the effects of Yiqi Huoxue Jiedu formula（YHJF） on immune cell exhaustion in the spleen of septic mice and to explore and validate its potential intervention targets. MethodsMice were randomly divided into the sham-operated， model， low-dose YHJF（4.1 g·kg^-1）， and high-dose YHJF（8.2 g·kg^-1） groups. Except for the sham-operated group， a cecal ligation and puncture（CLP） procedure was performed to establish a mouse sepsis model. The treatment groups received oral administration of the corresponding doses， while the sham-operated and model groups received an equal volume of physiological saline. After the intervention， the 7-day survival rate of each group was recorded， and spleen samples were collected 72 h post-intervention， and the spleen index was calculated. Terminal deoxynucleotidyl transferase deoxyuridine triphosphate（dUTP） nick end labeling（TUNEL） staining was used to detect apoptosis in spleen cells. Enzyme-linked immunosorbent assay（ELISA） was performed to measure the levels of interleukin（IL）-4 and IL-10 in the serum. Transcriptomics and metabolomics were used to screen for differentially expressed genes（DEGs） and differential metabolites in the spleen， followed by bioinformatics analysis to identify key targets. Real-time quantitative polymerase chain reaction（Real-time PCR）， flow cytometry， and multiplex immunofluorescence were used to verify the expressions of key genes and proteins. ResultsThe high-dose YHJF group significantly improved the 7-day survival rate of septic mice（P0.05）. Compared with the sham-operated group， the model group showed a significant increase in apoptosis of spleen cells and a decrease in the spleen index at 72 h post-modeling， with markedly elevated peripheral serum IL-4 and IL-10 levels（P0.01）. Compared with the model group， the high-dose YHJF group showed a reduction in apoptosis of spleen cells， an increase in the spleen index， and a significant decrease in peripheral serum IL-4 and IL-10 levels（P0.05）. Spleen transcriptomics identified 255 DEGs between groups， potentially serving as intervention targets for YHJF. Gene Ontology（GO） enrichment analysis revealed that DEGs were mainly involved in biological processes such as natural killer（NK） cell-mediated positive immune regulation， cell killing， cytokine production， positive regulation of innate immune cells， and interferon production. Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis showed that DEGs were mainly involved in cytokine-cytokine receptor interactions， viral protein interactions with cytokines and cytokine receptors， chemokine signaling pathway， and nuclear transcription factor-κB（NF-κB） signaling pathway. Protein-protein interaction（PPI） network analysis identified CD160， granzyme B（GZMB）， and chemokine ligand 4（CCL4） as key targets for YHJF in treating sepsis. Metabolomics identified 46 differential metabolites that were significantly reversed by YHJF intervention， and combined transcriptomics and metabolomics analysis identified 17 differential metabolites closely related to CD160. Pathway enrichment revealed that these metabolites were mainly involved in glycerophospholipid metabolism， arachidonic acid metabolism， glycosylphosphatidylinositol（GPI） anchor biosynthesis， linoleic acid metabolism， and α-linolenic acid metabolism pathways. Verification results showed that， compared with the sham-operated group， the model group exhibited significantly elevated CD160 mRNA expression level in the spleen， along with markedly decreased CCL4 and GZMB mRNA expression， and had a significant increase in CD160 expression on the surface of natural killer T（NKT） cells in the spleen（P0.01）. Compared with the model group， the high-dose YHJF group had a significant decrease in CD160 mRNA expression in the spleen， a significant increase in CCL4 and GZMB mRNA expressions. Further flow cytometry and immunofluorescence revealed that compared with the sham-operated group， CD160 expression on the surface of splenic NKT cells in the model group was significantly increased（P0.01）， while high-dose YHJF intervention significantly reduced CD160 expression（P0.01）. ConclusionYHJF may alleviate NKT cell exhaustion in sepsis by downregulating the expression of the negative co-stimulatory molecule CD160， and this regulatory effect is closely related to fatty acid metabolism pathways. This study provides new insights and targets for further exploration of strengthening vital Qi and detoxifying strategy to improve immune cell exhaustion in acute deficiency syndrome of sepsis.

Objective To analyze the predictive value of different obesity indicators on acute cerebral infarction in patients with transient ischemic attack (TIA). Methods A total of 310 patients with TIA admitted to Tangdu Hospital of Air Force Medical University from March 2022 to September 2025 were selected and classified into non-event group (276 cases) and cerebral infarction group (34 cases) based on acute cerebral infarction within 90 days. The basic data and obesity-related indicators [body mass index (BMI), waist-to-hip ratio, body adiposity index (BAI), conicity index (C-index) and a body shape index (ABSI were compared. Multivariate logistic regression analysis was utilized to screen the independent influencing factors. Receiver operating characteristic (ROC) curve was drawn to analyze the predictive efficiency. Results The age, proportions of concurrent hypertension and concurrent diabetes mellitus, ABCD2 score, waist-to-hip ratio, ABSI, C-index, BMI and BAI in the cerebral infarction group were older or higher than those in the non-event group, and the symptoms duration was longer (P<0.05). The above indicators were all related factors for acute cerebral infarction in patients with TIA within 90 days (P<0.05), and ABSI and ABCD² score were independent influencing factors (P<0.05). The prediction model was manifested as Logit (P)=− 4.205 + 0.482 × ABCD² score + 1.120 × ABSI × 100. ABSI and ABCD2 score had predictive value on the occurrence of acute cerebral infarction (P<0.05), and the predictive efficiency of combined model was higher than that of single indicator (P<0.05). Conclusion Among different obesity indicators, ABSI has the highest value on predicting recent acute cerebral infarction in TIA. The prediction model based on ABSI and ABCD2 score exhibits the best predictive efficiency.

Objective To explore the method for establishing a pig left lung orthotopic transplantation model. Methods A total of 4 male Yorkshire pigs weighing (40.0±2.5) kg underwent surgical procedures, including anesthesia, tracheal intubation, donor lung procurement, and recipient lung transplantation. Histological morphology and blood gas analysis of the transplanted lungs were assessed 2 hours after reperfusion to evaluate the histological changes and function of the transplanted lungs. Results The pigs underwent left lung in situ transplantation, effectively simulating conditions relevant to human lung transplantation. Two hours after the transplantation, arterial blood gas analysis showed that arterial partial pressure of oxygen was 155.4-178.6 mm Hg, arterial partial pressure of carbon dioxide was 53.1-62.4 mm Hg, and the oxygenation index was 310.8-357.2 mm Hg. Hematoxylin and eosin staining indicated a low degree of pulmonary edema and minimal cellular infiltration. Conclusion The pig left lung orthotopic transplantation model possesses strong operability and stability. Researchers can replicate this model according to the described methods and further conduct basic research and explore clinical translational applications.

Organoid technology has become a highly promising in vitro model in biomedical research, but its traditional culture relies on animal-derived basement membrane matrices, which suffer from batch-to-batch variability, undefined biochemical composition, and poorly controllable mechanical properties. Recent studies have confirmed that matrix stiffness, viscoelasticity, topography, and temporal mechanical dynamics play regulatory roles equally as critical as biochemical signals in stem cell self-organization, lineage specification, and functional maturation of organoids. This review systematically summarizes the progress in understanding how extracellular matrix (ECM) mechanical properties regulate organoid formation. Focusing on the YAP/TAZ factors and the integrin receptors, which engage in crosstalk with biochemical signals to synergistically determine cell fate. We then elaborate how specific mechanical parameters, such as stiffness, viscoelasticity, and topography, influence each distinct stage of organoid development, from initial aggregation and compaction, through proliferative expansion, to lineage-committed differentiation and terminal functional maturation. Appropriate stiffness facilitates efficient aggregation and symmetry breaking, while excessive rigidity impairs proliferation and accelerates cellular senescence. In hepatic organoid expansion where only stiffness matching liver tissue(~6.2 kPa) supported robust growth. Viscoelastic stress relaxation promotes integrin clustering, enhances focal adhesion maturation, and stimulates endogenous ECM deposition, thereby supporting self-organized crypt-villus architecture and basement membrane assembly. Notably, dynamic softening of hydrogels enables optimal crypt budding, whereas rapid stress relaxation permits nephron segment coiling and drives vascular organoid maturation toward arteriole differentiation, highlighting stress relaxation as a key temporal parameter. Topographical cues provide contact guidance that directs polarized outgrowth, lumen formation, and neuronal orientation, all of which are essential for achieving organotypic architecture. The embedded 3D printing of complex neural networks enables precise control over branching geometries, facilitating studies of morphogenesis under spatial constraints. The engineered mechanical microenvironments have been successfully applied to recapitulate disease phenotypes: stiff matrices induce fibrosis and epithelial-mesenchymal transition in liver organoids, promote chemoresistance in cholangiocarcinoma. Furthermore, we summarize engineering strategies for both natural and synthetic hydrogels aimed at creating mechanically defined culture platforms. Natural biomaterial modifications achieve spatial separation of mechanical constraints from biochemical support, enabling self-limited compressive stimulation within a defined time window. Synthetic polymers allow independent tuning of stiffness, ligand density, and viscoelasticity through adjustable crosslinking and dynamic non-covalent interactions, thus providing orthogonal control over multiple mechanical parameters. Dynamic microenvironments that mimic progressive stiffening as observed in development, tumor evolution, or fibrosis can be constructed using composites that undergo spontaneous structural transitions. In the future, critical challenges include deciphering the synergistic rules governing mechanical-biochemical interplay, developing spatiotemporally programmable hydrogels that enable on-demand modulation of local mechanics, establishing comprehensive databases of tissue-specific mechanical parameter windows, and advancing standardized high-throughput screening platforms to accelerate preclinical validation and clinical translation. Moreover, integrating real-time mechanical sensors and non-invasive imaging techniques would allow dynamic monitoring of intracellular forces and tissue-level stress during culture. Ultimately, this review aims to provide a theoretical foundation for deepening our understanding of mechano-biological coupling mechanisms and for rationally designing high-fidelity, physiologically relevant organoid culture platforms that fully harness the regulatory power of mechanical cues.

Ulcerative colitis （UC） is a chronic inflammatory bowel disease primarily affecting the colon and rectum. With a rising incidence， UC has aroused an increasingly serious public health concern. The etiology and pathogenesis of UC are complex， involving genetic susceptibility， intestinal barrier dysfunction， immune dysregulation， and environmental factors such as diet， smoking， and infection. Current UC therapies face limitations including persistent drug resistance， multiple side effects， and compromised immune function， necessitating the exploration of novel intervention strategies. In recent years， with the continuous advancement of research into UC mechanisms， the Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB） signaling pathway has been revealved as one of the classical pathways involved in inflammation. Being crucial for maintaining intestinal homeostasis， this pathway regulates inflammatory and immune responses by controlling gene transcription， thereby playing a significant role in the occurrence and progression of UC. However， systematic summarization within this field remains lacking. Traditional Chinese medicine （TCM） possesses unique advantages through its multi-target， multi-pathway， and multi-mechanism approaches， being capable of curbing the pathological progression of UC by targeting key molecules in the TLR4/NF-κB signaling pathway. This paper systematically summarizes the mechanisms by which TLR4/NF-κB-related pathways regulate UC， primarily encompassing inflammatory responses， oxidative stress， immune balance， and gut microbiota. It reviews single herbs （e.g.， Rosa roxburghii root and Saposhnikoviae Radix）， active components （e.g.， resveratrol， Tripterygium glycosides， and gypenosides）， herbal extracts （e.g.， Zingiberis Rhizoma extract and Taraxaci Herba extract）， and compound formulas for clearing heat and removing dampness， invigorating the spleen and warming the kidneys， clearing heat and detoxifying， harmonizing the liver and spleen， and treating cold and heat in complexity （e.g.， Huaihua Powder， Shenling Baizhu Powder， and Jiedu Huayu Formula）. The aim is to fully leverage the advantages of TCM in UC treatment and provide novel insights for clinical and pharmacological research on UC.

Ulcerative colitis （UC） is a chronic inflammatory bowel disease primarily affecting the colon and rectum. With a rising incidence， UC has aroused an increasingly serious public health concern. The etiology and pathogenesis of UC are complex， involving genetic susceptibility， intestinal barrier dysfunction， immune dysregulation， and environmental factors such as diet， smoking， and infection. Current UC therapies face limitations including persistent drug resistance， multiple side effects， and compromised immune function， necessitating the exploration of novel intervention strategies. In recent years， with the continuous advancement of research into UC mechanisms， the Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB） signaling pathway has been revealved as one of the classical pathways involved in inflammation. Being crucial for maintaining intestinal homeostasis， this pathway regulates inflammatory and immune responses by controlling gene transcription， thereby playing a significant role in the occurrence and progression of UC. However， systematic summarization within this field remains lacking. Traditional Chinese medicine （TCM） possesses unique advantages through its multi-target， multi-pathway， and multi-mechanism approaches， being capable of curbing the pathological progression of UC by targeting key molecules in the TLR4/NF-κB signaling pathway. This paper systematically summarizes the mechanisms by which TLR4/NF-κB-related pathways regulate UC， primarily encompassing inflammatory responses， oxidative stress， immune balance， and gut microbiota. It reviews single herbs （e.g.， Rosa roxburghii root and Saposhnikoviae Radix）， active components （e.g.， resveratrol， Tripterygium glycosides， and gypenosides）， herbal extracts （e.g.， Zingiberis Rhizoma extract and Taraxaci Herba extract）， and compound formulas for clearing heat and removing dampness， invigorating the spleen and warming the kidneys， clearing heat and detoxifying， harmonizing the liver and spleen， and treating cold and heat in complexity （e.g.， Huaihua Powder， Shenling Baizhu Powder， and Jiedu Huayu Formula）. The aim is to fully leverage the advantages of TCM in UC treatment and provide novel insights for clinical and pharmacological research on UC.

Objective : To investigate the status of electronic cigarette use and its influencing factors among residents aged ≥15 years in Chengdu City in 2021 and 2024, so as to provide the evidence for optimizing electronic cigarette control strategies. Methods: Two rounds of cross-sectional surveys were conducted in 2021 and 2024. Multi-stage cluster random sampling method was used to select permanent residents aged ≥15 years in Chengdu City as survey subjects. The Chinese Adult Tobacco Survey Questionnaire was used to collect information on gender, age, and electronic cigarette use. Complex sampling weighting was used to calculate the rate of electronic cigarette use and the proportion of reasons for electronic cigarette use. A complex sampling multivariable logistic regression model was used to analyze the influencing factors for electronic cigarette use. Results: A total of 7 950 and 6 920 people were surveyed in 2021 and 2024, respectively, including 3 868 and 3 653 males (48.65% and 52.79%), and 4 082 and 3 267 females (51.35% and 47.21%), respectively. The rate of electronic cigarette use in 2024 was lower than in 2021 (2.29% vs. 3.15%, P<0.05). In 2024, the rates of electronic cigarette use were lower in males, rural areas, people aged 15-<45 years, educational level of bachelor' s degree or above, occupation in government/public institutions or occupation in enterprises/commercial/service industries, and current smokers compared with 2021, while the rate of electronic cigarette use was higher in residents aged ≥65 years compared with 2021 (all P<0.05). In 2021 and 2024, the main reasons for electronic cigarette use were smoking cessation and the belief that electronic cigarettes are less harmful. Smoking cessation accounted for 27.78% and 23.28%, and the belief that electronic cigarettes are less harmful accounted for 24.26% and 22.10%, respectively. Complex sampling multivariable logistic regression analysis showed that age (15-<25 years, OR=15.682, 95%CI: 5.660-43.450; 25-<45 years, OR=7.463, 95%CI: 2.987-18.647; 45-<65 years, OR=3.425, 95%CI: 1.562-7.510), educational level (bachelor' s degree or above, OR=2.916, 95%CI: 1.444-5.891), and smoking (current, OR=23.992, 95%CI: 8.182-70.353; former, OR=20.393, 95%CI: 6.150-67.624) were influencing factors for electronic cigarette use in 2021. Gender (male, OR=2.279, 95%CI: 1.074-4.832), age (25-<45 years, OR=2.114, 95%CI: 1.148-3.892), educational level (high school/vocational high school/technical secondary school, OR=2.792, 95%CI: 1.414-5.514; bachelor' s degree or above, OR=3.321, 95%CI: 1.567-7.039), occupation (unemployed, retired, and retired, OR=0.449, 95%CI: 0.226-0.892), and smoking (current, OR=10.547, 95%CI: 5.563-19.994; former, OR=8.998, 95%CI: 3.833-21.125) were influencing factors for electronic cigarette use in 2024. Conclusions Compared with 2021, the rate of electronic cigarette use among residents in Chengdu City decreased in 2024. Young people, males, bachelor' s degree or above, and current smokers are the key populations for electronic cigarette control. The cognitive bias held by residents that electronic cigarette can aid in smoking cessation and pose minimal harm needs to be corrected.

Ulcerative colitis(UC)is a recurrent,chronic,nonspecific inflammatory bowel disease.Shaoyao Decoction has shown good efficacy in the treatment of UC,which mainly regulates the balance of cytokines to achieve the purpose of treating UC.This article reviewed the research progress in the mechanism of Shaoyao Decoction in the treatment of UC from two aspects of pro-inflammatory cytokines and anti-inflammatory cytokines.Research has shown that Shaoyao Decoction can regulate the body's immune response,alleviate intestinal inflammation,protect the intestinal mucosal barrier,restore normal structure and function of the colon mucosa through pro-inflammatory cytokines such as tumor necrosis factor-α,interferon-γ,interleukin(IL)-1β,IL-6,IL-8,IL-17,and anti-inflammatory cytokines such as transforming growth factor-β,IL-4,IL-10,IL-13,which can provide reference for UC related mechanism research and clinical treatment.

OBJECTIVE: To explore the protective effects and underlying mechanisms of H ₂S against lipid peroxidation-mediated carbonyl stress in the uranium-treated NRK-52E cells. METHODS: Cell viability was evaluated using CCK-8 assay. Apoptosis was measured using flow cytometry. Reagent kits were used to detect carbonyl stress markers malondialdehyde, 4-hydroxynonenal, thiobarbituric acid reactive substances, and protein carbonylation. Aldehyde-protein adduct formation and alcohol dehydrogenase, aldehyde dehydrogenase 2, aldo-keto reductase, nuclear factor E2-related factor 2 (Nrf2), and cystathionine β-synthase (CBS) expression were determined using western blotting or real-time PCR. Sulforaphane (SFP) was used to activate Nrf2. RNA interference was used to inhibit CBS expression. RESULTS: GYY4137 (an H ₂S donor) pretreatment significantly reversed the uranium-induced increase in carbonyl stress markers and aldehyde-protein adducts. GYY4137 effectively restored the uranium-decreased Nrf2 expression, nuclear translocation, and ratio of nuclear to cytoplasmic Nrf2, accompanied by a reversal of the uranium-decreased expression of CBS and aldehyde-metabolizing enzymes. The application of CBS siRNA efficiently abrogated the SFP-enhanced effects on the expression of CBS, Nrf2 activation, nuclear translocation, and ratio of nuclear to cytoplasmic Nrf2 and concomitantly reversed the SFP-enhanced effects of the uranium-induced mRNA expression of aldehyde-metabolizing enzymes. Simultaneously, CBS siRNA reversed the SFP-mediated alleviation of the uranium-induced increase in reactive aldehyde levels, apoptosis rates, and uranium-induced cell viability. CONCLUSION H ₂S induces Nrf2 activation and nuclear translocation, which modulates the expression of aldehyde-metabolizing enzymes and the CBS/H ₂S axis. Simultaneously, the Nrf2-controlled CBS/H ₂S axis may at least partially promote Nrf2 activation and nuclear translocation. These events form a cycle-regulating mode through which H ₂S attenuates the carbonyl stress-mediated NRK-52E cytotoxicity triggered by uranium.
NF-E2-Related Factor 2/genetics* ; Animals ; Hydrogen Sulfide/pharmacology* ; Rats ; Signal Transduction/drug effects* ; Lipid Peroxidation/drug effects* ; Cell Line ; Uranium/toxicity* ; Antioxidant Response Elements ; Kidney/metabolism* ; Oxidative Stress/drug effects* ; Cell Survival/drug effects* ; Apoptosis/drug effects*