Abstract:
Objective　To humanize the variable region of the IL9-2G1 rabbit monoclonal antibody （IL9 2G1 mAb） and to evaluate its biological activity.Methods　The humanized sequences of IL9-2G1 mAb were designed. The heavy chain variable region （VH） and light chain variable region （VL） of the antibody were humanized and cloned into vector backbones containing the constant region frameworks of human IgG1 and Ig Kappa subtypes. Protein purification was carried out using the Chinese hamster ovary cell （CHO） expression system. Enzyme-linked immunosorbent assay （ELISA） was used to qualitatively analyze the antigen-binding activity of the antibodies. After screening the optimal humanized antibody candidate， surface plasmon resonance （SPR） was performed to detect molecular affinity. Western blot was used to examine the effects of the rIL-9 group and the rIL-9 + hIL-9 mAb group on the phosphorylation of Janus kinase 1/signal transducer and activator of transcription 3 （JAK1/STAT3）， while ELISA was employed to assess the impact of the Th9 group and the Th9 + hIL-9 mAb group on interleukin-9 （IL-9） secretion.Results　Purification and ELISA results showed that the heavy chain OH and light chain FL exhibited good compatibility and expression performance. SPR results indicated that the equilibrium dissociation constant （KD） of M501042-OHFL against the analyte M30903 was 1.33×10-7 M， which was superior to the parental 2G1 antibody. The Western blot results showed that the levels of phosphorylated JAK1 （p-JAK1） and phosphorylated STAT3 （p-STAT3） were significantly decreased in the rIL-9 + hIL-9 mAb group compared with the rIL-9 group （P<0.01）. The ELISA results indicated that the secretion of IL-9 was significantly reduced in the Th9 + hIL-9 mAb group compared with the Th9 group （P<0.01）.Conclusion　Humanization modification is performed on the variable region of IL9-2G1 rabbit monoclonal antibody， and the humanized antibody hIL-9 mAb OHFL with high affinity and strong binding stability is successfully constructed， which provides an experimental basis for subsequent research.

Abstract:
Objective　To investigate the role of interleukin （IL）-8 secreted by tumor-associated macrophages （TAMs） in colorectal cancer （CRC） metastasis and its potential therapeutic value.Methods　The TAMs model was constructed by phorbol ester （PMA） combined with IL-4/IL-13， and the model phenotype was verified by flow cytometry， RT-qPCR and ELISA. Human CRC HCT116 cells were treated with conditioned medium （CM） from M0 macrophages and TAMs， and the metastatic capacity of HCT116 cells was evaluated by wound-healing assay and Transwell assay. The expression levels of IL-8 in M0 macrophages and TAMs were detected by Western blot， RT-qPCR and ELISA. The expression level of epithelial-mesenchymal transition （EMT） markers （E-cadherin， N-cadherin） in HCT116 cells treated with CM-M0 and CM-TAMs were detected by Western blot and RT-qPCR. HCT116 cells were stimulated with CM-TAMs supplemented with IL-8 receptor inhibitor Reparixin. The HCT116 cells metastasis ability was evaluated by wound-healing assay and Transwell assay， and the expression levels of E-cadherin and N-cadherin were detected by Western blot and RT-qPCR. Finally， a splenic injection liver metastasis model was established using murine CRC cells CT26-LUC， and the effect of Reparixin on this animal model was observed by in vivo imaging.Results　Flow cytometry showed that the proportion of M2 macrophages （CD11b+CD206+ cells） was elevated in the TAMs model. RT-qPCR and ELISA assays revealed that the levels of M2 macrophage markers IL-10 and TGF-βsignificantly increased. Wound-healing assay and Transwell assay demonstrated that CM-TAMs significantly enhanced the migration and invasion ability of HCT116 cells. Western blot， RT-qPCR and ELISA assays showed that TAMs highly expressed IL-8 and simultaneously induced downregulation of E-cadherin and upregulation of N-cadherin in HCT116 cells. Reparixin not only reversed the pro-metastatic effects of CM-TAMs， but also upregulated the expression level of E-cadherin and downregulated that of N-cadherin in HCT116 cells. Furthermore， it reduced the bioluminescence signal intensity of liver metastatic lesions in the splenic injection liver metastasis model.Conclusion　The TAMs promote CRC metastasis by secreting IL-8 to induce EMT. Inhibiting IL-8 secretion in TAMs or blocking the binding of IL-8 to its receptor may provide a new strategy for CRC treatment.

Abstract:
Objective　To investigate the role of disco interacting protein 2 homolog B （DIP2B） in the proliferation， migration， and invasion processes of gastric cancer cells.Methods　Bioinformatic analysis was performed using public datasets， including TCGA， GEPIA and TIMER2.0， to assess DIP2B expression and its clinical prognostic value in gastric cancer. DIP2B expression levels in gastric cancer cell lines were validated by Western blot and RT-qPCR. A DIP2B knockdown cell model was established via siRNA mediated gene silencing， and the knockdown efficiency was confirmed. Cell proliferation was assessed using real-time cellular analysis （RTCA） and colony formation assays. Cell migration and invasion were evaluated by Transwell assays， while flow cytometry was employed to detect changes in apoptosis and cell cycle distribution.Results　DIP2B was significantly upregulated in gastric cancer cells and correlated with poor prognosis in patients （P<0.05）. The experimental results revealed that DIP2B knockdown markedly suppressed the proliferation， migration， and invasion of gastric cancer cells， while promoting apoptosis and inducing cell cycle arrest （P<0.05）.Conclusion　DIP2B is highly expressed in gastric cancer and associated with unfavorable prognosis. Knockdown of DIP2B inhibits the proliferation， migration， and invasion of gastric cancer cells and promotes apoptosis， suggesting that DIP2B may serve as a potential prognostic biomarker and therapeutic target for gastric cancer.

Abstract:
Objective　To prepare photosensitive nanodiscs loaded with rapamycin and evaluate their in vitro antitumor activity against ovarian cancer.Methods　Nanodiscs were prepared by the thin-film hydration method， and their particle size， potential and morphology were characterized by dynamic light scattering and transmission electron microscopy. High performance liquid chromatography was established for the determination of rapamycin content， and the specificity， linearity， precision， repeatability， stability， and recovery rate of the method were verified. The singlet oxygen generation and light-controlled drug release properties of nanodiscs under near-infrared light were evaluated respectively by singlet oxygen detection probe and reverse dialysis method. The cytotoxicity of the nanodiscs against human ovarian cancer cells （OVCAR-8） was evaluated by detecting cell metabolic activity. Confocal microscopy and flow cytometry were used to investigate the cellular uptake and endocytosis mechanisms of the nanodiscs.Results　The nanodiscs exhibited a hydrodynamic diameter of （63.66±0.27） nm， a zeta potential of （-18.2 ± 2.4） mV， and demonstrated good stability. The results from transmission electron microscopy revealed that the nanodiscs exhibited spherical and short rod-like morphologies. The peak area of rapamycin showed good linearity in the concentration range of 0.2-40 μg/mL （R²=0.999 9）， and the relative standard deviations of precision， repeatability and recovery were all less than 5%. Under near-infrared light irradiation， the nanodiscs could effectively produce singlet oxygen. At the same time， the singlet oxygen generated could induce lipid membrane oxidation， which significantly promoted the responsive release of rapamycin （P<0.001）. Nanodiscs had no obvious cytotoxicity， but the cytotoxicity to ovarian cancer cells was significantly enhanced after near-infrared irradiation. Cellular uptake experiments confirmed that the nanodiscs mainly entered cells through clathrin-mediated， energy-dependent endocytosis （P<0.001）， with drug release triggered by light exposure.Conclusion　Rapamycin photosensitive nanodiscs were successfully prepared， which showed significant light-controlled release properties and enhanced anti-ovarian cancer activity in vitro.

Abstract:
Objective　To investigate whether the CD151 YXXφ motif point mutant affects vascular permeability by regulating vesicle internalization and recycling.Methods　CD151 knockout （KO） and wild-type （WT） mice were used. A modified Miles assay was performed to compare vascular permeability in the dorsal and postauricular skin of the two groups of mice under vascular endothelial growth factor A （VEGF-A） stimulation. The CD151 YXXφ motif point mutant YALA was constructed and transfected into human endothelial cells （EA.hy926） using an adenoviral vector. Multiple experimental groups were set up to detect the permeability of endothelial cell monolayers to FITC-dextran under baseline conditions and after VEGF-A stimulation. Western blot （WB）， RT-qPCR， and immunofluorescence were used to examine the expression and distribution of VE-cadherin. The internalization rate of VE-cadherin was quantitatively compared using a cell surface biotinylation assay. Phalloidin was used to label F-actin to observe changes in cytoskeletal tension.Results　The Miles assay showed that under VEGF-A stimulation， dye extravasation in the dorsal and postauricular skin of KO mice was significantly higher than that in WT mice （all P<0.05）. At 60 min and 120 min after VEGF-A stimulation， the permeability of endothelial cell monolayers overexpressing the YALA mutant to FITC-dextran significantly increased （P<0.05）. WB and RT-qPCR results indicated that the YALA mutant did not affect the total protein or mRNA expression levels of VE-cadherin. Immunofluorescence revealed that the YALA mutant disrupted the continuous distribution of VE-cadherin on the cell membrane and its perinuclear co-localization with CD151 under VEGF-A stimulation. The cell surface biotinylation assay confirmed that the YALA mutant significantly reduced the internalization rate of VE-cadherin （P<0.05）. Cytoskeletal staining showed that the YALA mutant caused the formation of thick tension fibers in the actin cytoskeleton of endothelial cells， indicating cell contraction.Conclusion　The CD151 YXXφ motif point mutation affects vesicle internalization and recycling in endothelial cells， influences the expression and internalization of VE-cadherin， and consequently affects vascular permeability.

Abstract:
Objective　To explore the structural characteristics， biocompatibility， anti-inflammatory and antioxidant properties of hyaluronic acid （HA）/silk fibroin （SF） composite hydrogel scaffolds loaded with epigallocatechin gallate （EGCG）， and to provide a new material basis for the treatment of periodontitis.Methods　The composite hydrogel scaffolds were constructed by cross-linking HA and SF and loaded with EGCG. The water absorption rate， degradation property and material characteristics were detected by scanning electron microscopy. The cytocompatibility and anti-inflammatory and antioxidant properties were evaluated by in vitro experiments.Results　The EGCG-loaded scaffolds exhibited a porous network structure and desirable swelling properties. In vitro experiments demonstrated that the scaffolds possessed good biocompatibility， along with the ability to inhibit the expression of inflammatory factors and scavenge reactive oxygen species.Conclusion　The HA/SF-EGCG composite scaffolds have excellent biocompatibility and dual functional activities of anti-inflammatory and antioxidant， providing a new strategy for improving the clinical treatment effect of periodontitis.

Abstract:
Objective　To explore the function and related molecular mechanisms of magnoflorine against colon cancer via network pharmacology， molecular docking， and in vitro cell experiments.Methods　In this study， the canonical SMILES of magnoflorine was obtained from the PubChem database， and the potential targets of magnoflorine were predicted by the Swiss Target Prediction database， while the disease targets of colon cancer were obtained from the DisGeNET， GeneCards and OMIM databases. The intersecting targets between magnoflorine's predicted targets and colon cancer disease targets were taken， and a protein-protein interaction （PPI） network was constructed and analyzed. The DAVID online database was employed to conduct Gene Ontology （GO） and KEGG pathway enrichment analyses on core targets. The top five key targets screened were docked with magnoflorine using AutoDock software. Finally， cellular experiments including CCK-8 assays， EdU experiments， cell scratch assays， and Transwell assays were conducted to validate the results from network pharmacology and molecular docking.Results　44 key targets of magnoflorine in resisting colon cancer were acquired. The molecular docking results showed that magnoflorine had a strong binding activity with the core target signal transducer and activator of transcription 3 （STAT3） in the top five of the PPI network. Cellular experiments confirmed that magnoflorine could inhibit the proliferation and migration of colon cancer cells by suppressing the JAK/STAT3 signaling pathway.Conclusion　Magnoflorine may inhibit the proliferation and migration of colon cancer cells by regulating the JAK/STAT3 signaling pathway.

Abstract:
Objective　To investigate the therapeutic mechanism of artemether in diabetic sarcopenia（DS）using network pharmacology and animal experiments.Methods　Potential active components and therapeutic targets were screened using artemisinin as the parent compound. The predicted targets were intersected with DS-related targets， followed by construction of a protein-protein interaction （PPI） network. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses were performed to identify the key biological processes and signaling pathways. Molecular docking was further used to evaluate the binding affinity between artemether and core targets. The DS mouse model was established using db/db mice， followed by artemether intervention. Fasting blood glucose， body weight， diabetes-related symptoms， body composition， grip strength， serum and skeletal muscle triglyceride levels， and muscle fiber cross-sectional area （CSA） were assessed. The expression levels of forkhead box O1 （FoxO1）， Atrogin-1， muscle RING finger protein 1 （MuRF1）， acyl-CoA synthetase short-chain family member 2 （ACSS2）， carnitine palmitoyltransferase 2 （CPT2）， and fatty acid-binding protein 3 （FABP3） in skeletal muscle were detected by qRT-PCR， Western blot， immunofluorescence， and immunohistochemistry. In addition， metabolomics was performed to analyze changes in acylcarnitine metabolites in skeletal muscle.Results　A total of 68 overlapping targets between artemether and diabetic sarcopenia （DS） were identified. The core targets included AKT1， FoxO1， NFKB1， FBXO32， and TRIM63， which were mainly enriched in the phosphoinositide 3-kinase/protein kinase B （PI3K-Akt） signaling pathway， forkhead box O （FoxO） signWaling pathway， tumor necrosis factor （TNF） signaling pathway， and insulin resistance-related pathways. Molecular docking analysis showed that artemether exhibited favorable binding affinity with the core targets. Animal experiments demonstrated that artemether reduced fasting blood glucose， ameliorated metabolic symptoms， increased lean mass and grip strength， decreased serum and skeletal muscle triglyceride levels， and increased muscle fiber cross-sectional area （CSA） in DS mice. In addition， artemether downregulated the mRNA and protein expression levels of FoxO1， FBXO32/Atrogin-1， TRIM63/MuRF1， ACSS2， CPT2， and FABP3， and improved the disturbance of acylcarnitine metabolism in skeletal muscle.Conclusion　Artemether improves metabolic and skeletal muscle phenotypes in DS mice. Its effects may be associated with the amelioration of lipid metabolic disorders and the downregulation of FoxO1 and its downstream ubiquitin-proteasome pathway-related factors Atrogin-1 and MuRF1 in skeletal muscle.

Abstract:
Objective　To investigate the expression level of mammalian sterile 20-like kinase 1 （MST1） in prostate cancer （PCa） tissues， its biological functions， and its regulatory effects on androgen receptor （AR） and vascular endothelial growth factor （VEGF） expression.Methods　Immunohistochemistry was performed to detect MST1 expression in 77 prostate cancer tissues and 34 benign prostatic hyperplasia （BPH） tissues， and the relationship between MST1 expression and clinicopathological parameters was analyzed. MST1-overexpressing C4-2 and LNCaP cell lines were constructed. CCK-8 assay， Transwell migration assay， and angiogenesis assay were conducted to evaluate the effects of MST1 on cell proliferation， migration， and angiogenesis. The expression changes of AR and VEGF were detected by qRT-PCR and Western blot.Results　MST1 expression was significantly lower in PCa tissues compared with BPH tissues （P=0.012）. Low MST1 expression was significantly associated with tumor metastasis （P=0.033）. MST1 overexpression inhibited C4-2 cell proliferation and migration， with transmembrane migrated cells reduced by 53.6% （P<0.01）. MST1 overexpression decreased the mRNA and protein expression levels of AR and VEGF （P<0.05）. Angiogenesis assay demonstrated that the length of new vessel structure was significantly reduced （P<0.05）.Conclusion　MST1 shows a low expression level in PCa tissues and is associated with tumor metastasis. MST1 exerts tumor-suppressive effects in prostate cancer by inhibiting cell proliferation， migration， and angiogenesis， as well as downregulating the expression of AR and VEGF， indicating its potential as a therapeutic target for prostate cancer.

Abstract:
Objective　To establish and characterize an inducible， CD4+T cell-specific ablation mouse model （CD4-iDTR） using the Cre-loxP and diphtheria toxin receptor （DTR） systems， and to evaluate its depletion efficacy and specificity.Methods　CD4-iDTR mice were generated through cross-breeding， with genotypes confirmed via PCR-based screening. Mice were categorized into an untreated （control） group and a diphtheria toxin （DT） induction group. Flow cytometry was employed to compare the baseline distribution of CD4+ T cells between untreated CD4-iDTR and wild-type C57BL/6 mice.In the DT group， CD4+ T cell ablation was induced by subcutaneous DT administration. The depletion efficiency in skin tissues and skin-draining lymph nodes （sdLNs） was quantified at 24 h and 1 week post-injection. Furthermore， the proportions of neutrophils， macrophages， dendritic cells， and other CD11b+ immune cell subsets in the skin were monitored to assess the model specificity.Results　Genomic profiling confirmed the successful generation of the CD4-iDTR line. Under steady-state conditions， no significant differences were observed in the frequency or absolute number of CD4+ T cells in the skin and sdLNs between CD4-iDTR and C57BL/6 mice （P>0.05）. Following DT induction， the CD4+ T cell population in the skin exhibited decline within 24 h and 1 week （P<0.05）， whereas the CD4+ T cell counts in sdLNs remained stable. The percentages of cutaneous neutrophils， macrophages， and other CD11b+ myeloid populations were not significantly altered following DT treatment （P>0.05）.Conclusion　The CD4-iDTR mouse model is successfully established and bred， achieving efficient， specific， and inducible ablation of cutaneous CD4+ T cells without significant interference with the skin immune microenvironment. This model serves as an ideal experimental tool for investigating skin-specific immunological mechanisms.

Abstract:
Objective　To investigate the protective effect of liver-targeted fibroblast growth factor 21 （FGF21） overexpression against CCl₄-induced acute liver injury （ALI） and its association with silent information regulator 1/glutathione peroxidase 4 （SIRT1/GPX4） expression and ferroptosis-related markers.Methods　Female ICR mice （8 weeks old） were randomly divided into Control， CCl₄， adeno-associated virus （AAV）-FGF21， and AAV-FGF21 + CCl4 groups. Liver-targeted FGF21 overexpression was achieved via tail vein injection of AAV8-FGF21 virus with green fluorescent protein （GFP） labeling. ALI was induced by intraperitoneal injection of CCl₄ （0.3 mL/kg）， with sample collection at 12 hours. Body weight and liver weight were recorded to calculate the liver weight/body weight. Alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured using a biochemical analyzer. FGF21 expression in liver tissues was detected by immunofluorescence. Hepatic histopathology was evaluated by HE staining. RT-qPCR was performed to assess mRNA expression of ferroptosis and iron metabolism-related genes （ACSL4， PTGS2， HJV， FPN， GPX4）， FGF21， and SIRT1. Protein expression of FGF21， SIRT1， and GPX4 were determined by Western blot. Iron content， malondialdehyde （MDA）， glutathione （GSH）， and superoxide dismutase （SOD） activity in liver homogenate were quantified using assay kits.Results　Compared with the Control group， the FGF21 mRNA and protein expression elevated in CCl4 group； serum ALT/AST levels and liver weight/body weigh increased. The HE staining results showed that the liver cell membranes were ruptured. The mRNA levels of ACSL4 and PTGS2， the iron content and MDA levels all increased （P<0.01）. The mRNA levels of SIRT1， GPX4， HJV， and FPN， the GSH level and SOD activity all decreased （P<0.05）， and the protein levels of SIRT1 and GPX4 both decreased （P<0.05）. Compared with the CCl4 group， the serum levels of ALT and AST in the AAV-FGF21 + CCl4 group were decreased （P<0.01）， and the liver/body ratio of mice decreased （P<0.01）. The results of HE staining showed that the degree of liver injury was alleviated. The mRNA levels of ACSL4 and PTGS2， iron content and MDA level all decreased （P<0.01）， while the mRNA levels of SIRT1， GPX4， HJV， FPN， GSH level and SOD activity all increased （P<0.05）. The protein levels of SIRT1 and GPX4 both increased （P<0.05）.Conclusion　FGF21 overexpression is associated with enhanced SIRT1/GPX4 expression and suppression of ferroptosis features， thereby contributing to the alleviation of CCl4-induced ALI.

Abstract:
Objective To construct an interleukin-9（Il9）gene knockout mouse model on a C57BL/6N back⁃ ground，establish a reliable genotyping method，and provide a standardized animal model for investigating Il9-re⁃ lated immune mechanisms and diseases. Methods Il9 knockout mouse strain（C57BL/6N-Il9em1Cya）was generated using conventional gene knockout technology. The method for DNA extraction from mouse tails was optimized to en ⁃ sure template quality. Two sets of specific primers（F1/R1 and F1/R2）were designed to amplify target fragments by PCR，and mouse genotypes were determined by agarose gel electrophoresis. Specifically，primer pair F1/R1 tar⁃ geted the knockout sequence with an expected product of 465 bp，while F1/R2 targeted the wild-type sequence with an expected product of 703 bp. Subsequently，Western blot was performed to verify the knockout efficiency of Il9 in primary immune cells and key organs of the offspring mice，and flow cytometry was used to assess the effects of Il9 deficiency on the function of the immune system in major immune cell populations. Results Il9 knockout mice were successfully bred and genotyped. PCR identification showed that homozygous mice（-/-）exhibited only a 465 bp band，wild-type mice（+/+）displayed only a 703 bp band，and heterozygous mice（+/-）showed both bands si⁃ multaneously. Meanwhile，Western blot analysis revealed significantly decreased IL-9 expression in various organs of the offspring mice（P<0. 001）. Flow cytometric results demonstrated that Il9 deficiency exerted no obvious ef⁃ fects on the proportions of peripheral blood lymphocytes，splenic lymphocytes，and peritoneal macrophages. Con ⁃ clusion An Il9 gene knockout mouse model on a C57BL/6N background is successfully constructed. The estab⁃ lished PCR genotyping system is highly efficient and accurate，which can effectively distinguish mice of different genotypes. This model provides a stable and reliable experimental basis for subsequent functional studies of Il9 and exploration of the mechanisms underlying Il9-related diseases.

Abstract:
Objective　To systematically screen and validate key immunogenic cell death （ICD）-related genes that play important roles in primary sjögren’s syndrome （pSS） using multi-omics data.Methods　Gene expression profiles of pSS patients and healthy controls （GSE66795， 131pSS patients/29 healthy controls） were obtained from the Gene Expression Omnibus （GEO） database. Candidate genes were selected by intersecting these profiles with known ICD core gene sets and pSS-related targets. The CIBERSORT algorithm was used to analyze immune cell infiltration characteristics. Validation of gene expression was performed using transcriptomic data from sorted CD8⁺ T cell subsets （GSE93683）. Finally， labial gland tissues and peripheral blood samples from 10 pSS patients and 4 healthy controls were collected， and key genes were experimentally validated by reverse transcription quantitative polymerase chain reaction （RT-qPCR）.Results　Seven intersecting genes from the ICD-differentially expressed genes （DEGs）-pSS intersection were identified ［caspase 1 （CASP1）， forkhead box P3 （FOXP3）， interferon-gamma （IFNG）， myeloid differentiation primary response 88 （MYD88）， toll-like receptor 7 （TLR7）， DExD/H-box helicase 58 （DDX58）， and interferon induced helicase C domain 1 （IFIH1）］. Immune infiltration analysis showed that in pSS labial gland tissues， the proportions of naïve B cells and dendritic cells significantly increased， while the proportions of resting NK cells and M0 macrophages decreased （P<0.05）. Transcriptomic data of CD8⁺T cell subsets indicated that IFIH1 expression was significantly elevated in central memory CD8⁺T cells （P<0.05）. RT-qPCR experimental validation revealed that DDX58， IFIH1， and CASP1 were significantly upregulated in both peripheral blood leukocytes and labial gland tissues of pSS patients （all P<0.05）， whereas IFNG expression increased in peripheral blood but decreased in labial gland tissues （P<0.05）.Conclusion　This study identified and confirmed DDX58， IFIH1， CASP1， and IFNG as key ICD-related genes in pSS through a combination of bioinformatics and experimental validation， providing new candidate targets for further understanding the immunopathological mechanisms of pSS.

Abstract:
Objective　To investigate the expression patterns of key steroidogenic enzymes （CYP11A1， HSD3B1， and CYP19A1） in placental tissues from preeclampsia （PE） patients and evaluate​ their potential as biomarkers for PE.Methods　A case-control study was conducted using placental tissues from PE patients （PE group， n=3） and normotensive pregnant women （control group， n=3）. RNA sequencing was performed to identify differentially expressed genes （DEGs）， followed by validation of target gene expression using Western blot， qRT-PCR， and immunohistochemistry. Steroid hormone levels in placental tissues and serum were measured by ELISA. Spearman correlation analysis was used to assess associations between protein expression and clinical parameters， and receiver operating characteristic （ROC） curve analysis was performed to evaluate the predictive value of target genes for PE.Results　The results showed that 501 DEGs （P<0.05 and |Log2Fold Change| >1） were identified in PE placental tissues， with KEGG enrichment analysis revealing significant involvement in steroid hormone biosynthesis pathways. Compared with the control group， PE placental tissues exhibited significantly decreased protein and mRNA expression of HSD3B1 and CYP19A1 （P<0.01）， while CYP11A1 expression was upregulated （P<0.05）. Serum estradiol （E2） and progesterone （P4） levels were significantly reduced in the PE group （P<0.01）， whereas testosterone （T） levels were elevated （P<0.05）. Correlation analysis demonstrated that CYP11A1 expression was positively associated with systolic blood pressure （SBP）， diastolic blood pressure （DBP）， and soluble fms-like tyrosine kinase-1 （sFlt-1） （P<0.01）， but negatively correlated with placental growth factor （PLGF） （P<0.01）. Conversely， HSD3B1 and CYP19A1 expression showed inverse correlations with these clinical parameters （P<0.01）. ROC analysis revealed that CYP11A1， HSD3B1， and CYP19A1 each had an area under the curve （AUC） of 0.778 for predicting PE， with 95%CI ranging from 0.614 to 0.942 （P=0.004）. The sensitivity was 94.4%， and the specificity was 66.7%.Conclusion　These findings suggest that dysregulated expression of steroidogenic enzymes in PE placental tissues contributes to hormonal imbalance， thereby playing a role in PE pathogenesis. Furthermore， CYP11A1， HSD3B1， and CYP19A1 may serve as potential diagnostic biomarkers for PE.

Abstract:
Objective　To investigate the metabolic mechanisms of Limosilactobacillus reuteri （L. reuteri） in alleviating chemotherapy-induced intestinal mucosal injury （CIMI）.Methods　C57BL/6 mice were randomly divided into the Control group， Model group， and L. reuteri treatment group （n=8 per group）. Mice in the L. reuteri group were intragastrically administered L. reuteri （1 × 109 CFU/mL） for 10 consecutive days. From day 5， mice in the Model and L. reuteri groups were intraperitoneally injected with 5-FU （65 mg/kg） for 6 consecutive days to establish a CIMI model. Body weight changes， colon length， and related indicators were recorded. Intestinal histopathological injury was assessed by HE staining with measurement of villus height and crypt depth. The expression levels of zonula occludens-1 （ZO-1）， Occludin， Claudin-4， Muc1， and Muc2 in ileal tissues were determined by RT-qPCR and immunohistochemistry to evaluate intestinal barrier function. In addition， untargeted metabolomics was performed on cecal contents from each group to analyze metabolic profiles and enrichment of differential metabolic pathways.Results　L. reuteri intervention significantly alleviated body weight loss （P<0.001） and colon shortening （P<0.05） in CIMI mice， effectively reducing ileal tissue pathological damage， and upregulating the expression of ZO-1， Occludin， Claudin4， Muc1， and Muc2 （P<0.05）. Untargeted metabolomics analysis showed that L. reuteri treatment reversed the abnormal metabolic profile in the cecal contents of CIMI mice， bringing it closer to normal. Various differential metabolites， such as acylcarnitines and polyunsaturated fatty acids， might play an important role in L. reuteri-mediated alleviation of CIMI. These differential metabolites were primarily enriched in key pathways such as linoleic acid metabolism， tryptophan metabolism， and unsaturated fatty acid biosynthesis.Conclusion　This study reveals that L. reuteri improves intestinal barrier function and alleviates CIMI by regulating the intestinal metabolic profile， particularly lipid and amino acid metabolism. It provides a novel potential strategy for the prevention and treatment of CIMI.

Abstract:
Objective　To investigate the protective effect of magnolol ester derivative YW against cisplatin （Cis）-induced acute kidney injury （AKI） and to explore its effect and preliminary mechanisms on Cis-induced AKI.Methods　The research group designed and synthesized the magnolol ester derivative YW using magnolol as the parent core. Molecular docking was employed to predict the binding interaction between YW and lysine acetyltransferase 7 （KAT7）. A Cis-induced AKI model was established in C57BL/6J mice. Mice were divided into the normal control group， model group， YW low/medium/high dose groups （17.5， 35， 70 mg/kg）， and the positive control amifostine group. Renal function was assessed by measuring serum creatinine （Scr） and blood urea nitrogen （BUN） levels. Pathological changes were evaluated via HE staining and scoring of kidney tissues. Protein expression levels of kidney injury molecule 1 （KIM-1）， lysine acetyltransferase 7 （KAT7）， and cyclin-dependent kinase inhibitor 1A （p21） in kidney tissues were detected by Western blot （WB）. HK-2 cells were treated with different concentrations of YW； cell viability was measured by CCK-8 assay. WB was used to detect the expression levels of the renal injury marker KIM-1， KAT7， and its downstream protein p21. Cellular senescence was assessed by senescence-associated β-galactosidase （SA-β-Gal） staining.Results　YW could stably bind to KAT7. In vivo experiments showed that， compared with the model group， all YW dose groups， especially the medium and high doses， significantly reduced the elevated levels of Scr and BUN in AKI mice （P<0.001） and effectively ameliorated renal histopathological damage. YW significantly downregulated the protein expression levels of KIM-1， KAT7， and p21 in kidney tissues （P<0.05）. Concentrations of YW≤10 μmol/L had no significant effect on HK-2 cell viability. YW inhibited the Cis-induced decline in HK-2 cell viability （P<0.05）， suppressed the Cis-induced upregulation of KIM-1， KAT7， and p21 proteins （P<0.05）， and significantly decreased the percentage of SA-β-Gal-positive cells （P<0.01）.Conclusion　YW suppresses the expression of KAT7 and p21 proteins and inhibits the decline in HK-2 cell viability in cisplatin-induced AKI， significantly mitigates HK-2 cell senescence， and improves renal function， providing preliminary experimental evidence for developing AKI treatment strategies based on structural optimization of natural products.

Abstract:
Objective　To reveal the effects of overexpression and knockdown of CDH18 gene on cervical cancer proliferation and migration levels through the Wnt/β-catenin signaling pathway in cervical cancer cells.Methods　A comprehensive analysis of CDH18-interacting molecules was performed via the bioinformatics STRING database （http：//cn.string-db.org）. Molecular docking between Wnt/β-catenin inhibitor XAV939 and β-catenin was performed using the bioinformatics tool AutoDock （http：//autodock.scripps.edu）. The effect of CDH18 on β-catenin subcellular distribution was detected by laser confocal microscopy. EdU assay， wound healing assay and Transwell assay were used to detect the effects of CDH18 gene overexpression， CDH18 gene knockdown and the XAV939 on cell proliferation and migration. Western blot was performed to measure the expression levels of Wnt/β-catenin pathway-related molecules， including β-catenin， transcription factor 4 （TCF4）， and cellular myelocytomatosis oncogene （c-Myc）， following CDH18 gene overexpression， CDH18 gene knockdown， or XAV939 treatment.Results　In SiHa and HeLa cells， CDH18 gene overexpression significantly increased the protein expression levels of β-catenin， TCF4 and c-Myc， decreased the protein expression of GSK-3β， and enhanced cell proliferation and migration， with statistically significant differences （P<0.05）. Conversely， CDH18 gene knockdown significantly reduced the protein expression of β-catenin， TCF4 and c-Myc， elevated GSK-3β expression， and suppressed cell proliferation and migration （P<0.05）.Treatment with XAV939 in SiHa and HeLa cells downregulated β-catenin， TCF4 and c-Myc， upregulated GSK-3β， and inhibited cell proliferation and migration， with significant differences （P<0.05）.Conclusion　CDH18 promotes the proliferation and migration of cervical cancer cells via the Wnt/β-catenin signaling pathway.

Abstract:
Objective　To explore the correlation between the polymorphisms of the solute carrier family7 member11 （SLC7A11） gene and the risk of non-cardia gastric cancer （NCGC） as well as the infection of Helicobacter pylori （H.pylori）.Methods　A total of 435 patients with NCGC and 479 healthy individuals undergoing physical examinations were included. The polymorphisms of the SLC7A11 gene （rs7674870， rs11734488， rs12506138， rs10022036） were genotyped using the polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） technique. The influence of each locus on H. pylori infection and the occurrence of NCGC was analyzed.Results　No significant associations were found among SNP rs7674870， rs11734488， and rs12506138 and H. pylori infection （P>0.05）. The CT genotype and CC genotype of rs12506138， when in a codominant model， reduced the risk of developing NCGC compared to the TT genotype； the A-C-C and A-T-C haplotypes constructed by SLC7A11 rs7674870， rs11734488， rs12506138 were associated with a reduced risk of NCGC； the A-T-T and G-C-C haplotypes were associated with an increased risk of NCGC.Conclusion　SLC7A11 rs7674870 and rs11734488 are not pivotal genetic loci for H. pylori infection and the pathogenesis of NCGC. SLC7A11 rs12506138 acts as a protective genetic locus for NCGC and antagonizes its oncogenic progression. The haplotype composed of the above three loci participates in regulating the overall genetic susceptibility to NCGC.

Abstract:
Objective　To investigate the relationship between social support and fertility quality of life in male infertility patients， and to analyze the parallel mediating roles of mindfulness and total sperm motility， thereby providing a theoretical basis for interventions aimed at improving fertility quality of life in this population.Methods　This study conducted a survey using anonymized electronic questionnaires among 680 infertile men from 2021 to 2024. The social support rating scale， mindful attention awareness scale， and fertility quality of life scale were used to assess social support， mindfulness， and fertility quality of life， respectively. Total sperm motility was evaluated through routine semen analysis. Pearson correlation analysis was performed to examine the relationships among scale scores， and the PROCESS macro was used to analyze the mediating effects of mindfulness and total sperm motility between social support and fertility quality of life.Results　Pearson correlation analysis showed that The fertility quality of life score was significantly positively correlated with the social support score （r=0.350）， subjective hosting score （r=0.279）， objective support score （r=0.274）， support utilization score （r=0.206）， mindfulness score （r=0.434）， and total sperm motility level （r=0.155） （P<0.05）. Mediation analysis demonstrated that both mindfulness and total sperm motility partially mediated the relationship between social support scores and fertility quality of life， with mediation effect proportions of 28.42% and 2.73%， respectively. Additionally， mindfulness scores and total sperm motility partially mediated the relationship between objective support （a subdimension of social support） and fertility quality of life， accounting for 28.78% and 3.70% of the effect， respectively. However， total sperm motility did not show significant mediation effects between subjective support， support utilization （other social support subdimensions）， and fertility quality of life.Conclusion　Mindfulness and total sperm motility are potential mediating factors through which social support influences fertility quality of life in male infertility patients. Clinically， mindfulness-based interventions to cultivate positive psychological traits， along with strategies to improve sperm motility， may enhance fertility quality of life in this population.

Abstract:
Objective　To investigate the relationship between periodontitis severity and brain functional indices-amplitude of low-frequency fluctuations （ALFF） and fractional ALFF （fALFF） in cognitively normal middle-aged and older adults， and to examine their associations with cognitive performance.Methods　42 cognitively normal middle-aged and older participants were enrolled. Periodontal data—including the number of remaining teeth （NTP）， bleeding on probing positivity rate （BOP%）， probing depth （PD）， and clinical attachment loss （CAL） as well as resting-state functional magnetic resonance imaging data were collected. Participants were divided into a mild periodontitis group and a moderate-to-severe periodontitis group according to periodontitis severity. Group differences in ALFF and fALFF were compared， and correlation analyses were performed.Results　Compared with the mild periodontitis group， the moderate-to-severe group showed decreased ALFF values in the right lingual gyrus， superior occipital gyrus， paracentral lobule， and in the left calcarine cortex as well as the precentral and postcentral gyri. Decreased fALFF values were observed in the right lingual gyrus， cuneus， and left calcarine cortex. Indicators related to periodontitis progression ［BOP%， PD=4-5 mm （%）， PD≥6 mm （%）， CAL=3-4 mm （%） and CAL≥5 mm （%）］ were negatively correlated with ALFF， fALFF and cognitive performance， whereas indicators reflecting better periodontal function ［PD=1-3 mm （%） and CAL=0-2 mm （%）］ were positively correlated with these measures.Conclusion　In cognitively normal middle-aged and older adults， periodontitis severity is associated with alterations in brain functional activity and cognitive performance.

Abstract:
Objective　To develop and validate a diagnostic prediction model for differentiating pulmonary tuberculosis from community-acquired pneumonia based on routine blood parameters.Methods　A total of 642 patients with pulmonary tuberculosis and 503 patients with community-acquired pneumonia were retrospectively enrolled from the Seventh Hospital of Tangshan. They were randomly divided into a training set and an internal validation set at a 7∶3 ratio. Additionally， 218 patients from the 981st Hospital were independently included as an external validation set. The Boruta algorithm and recursive feature elimination were employed to select predictors from 82 blood parameters， sex， and age. A multivariate Logistic regression model was established and evaluated using receiver operating characteristic （ROC） curves， calibration curves， and decision curve analysis.Results　The final model incorporated eight predictors， namely uric acid， urea nitrogen， alkaline phosphatase， monoamine oxidase， alanine aminotransferase， glutathione， neutrophil percentage， and age. The model achieved areas under the curve （AUCs） of 0.800 （95%CI： 0.769-0.830）， 0.787 （95%CI： 0.738-0.836）， and 0.736 （95%CI： 0.667-0.835） in the training， internal validation， and external validation sets， respectively. The model demonstrated good calibration， and decision curve analysis showed clinical net benefit within the threshold probability range of 10%-80%.Conclusion　The developed model exhibits good discriminative ability and clinical utility， serving as an effective early screening tool for primary healthcare institutions.

Abstract:
Erythropoietin-producing hepatoma receptor A2 （EphA2） is a member of the erythropoietin-producing hepatoma （Eph） subfamily of receptor tyrosine kinases. As a critical regulator of cancer development and progression， EphA2 can promote tumor initiation， growth， invasion， metastasis， stemness， and angiogenesis by activating or enhancing the oncogenic signaling pathways. High expression of EphA2 in tumor tissues is closely associated with poor prognosis， thus it is a very promising potential target for cancer treatment. This article reviews the molecular mechanisms by which EphA2 promotes cancer development and progression， aiming to provide clues and theoretical basis for EphA2-targeted cancer treatment.

Abstract:
Thalamic hemorrhage， a severe form of spontaneous intracerebral hemorrhage （ICH）， accounts for 6% to 25% of all ICH cases. It is associated with high mortality， imposing a substantial burden on both patients and society. Current research on the surgical management of thalamic hemorrhage primarily focuses on the indications， technical nuances， clinical outcomes， and complications of various surgical approaches. Stereotactic minimally invasive surgery for small-volume thalamic hemorrhage has been proven to improve patient prognosis， particularly for localized hematomas with a volume less than 15 mL. However， a systematic and standardized evaluation of the efficacy across different surgical methods is still lacking. This review analyzes existing literatures to explore different surgical approaches for thalamic hemorrhage， including craniotomy， stereotactic minimally invasive evacuation， and neuroendoscopic evacuation， assessing their respective advantages and limitations. The analysis reveals that traditional craniotomy is being used less frequently in the treatment of deep-seated thalamic hemorrhage due to its significant invasiveness. Stereotactic minimally invasive surgery demonstrates advantages in effectively improving neurological outcomes and reducing mortality for small to moderate hematomas， while neuroendoscopic surgery offers unique value in evacuating intraventricular hematomas and reducing shunt dependency. Additionally， the timing of surgery has a significant impact on patient prognosis， although the optimal timing remains to be further clarified. Current evidence is largely derived from retrospective studies， highlighting the need for high-quality prospective research to provide a more robust foundation for clinical decision-making.