Objective To investigate the expression of CDC40 in colorectal cancer tissues and its correlation with the clinicopathological features,so as to provide a basis for the early diagnosis,targeted therapy and prognostic assessment of colorectal cancer. Methods Immunohistochemical staining was used to evaluate the expression level of CDC40 in the tissues of 70 colorectal cancer patients,and the relationship between the CDC40 expression and the clinicopathological features was analyzed.The R software package was used for survival curve and multivariate prognostic analysis.Cell culture,transfection of CDC40 plasmid,proliferation experiments,Western blotting and nude mouse tumorigenesis experiments were conducted by using the colorectal cancer cell lines SW620 and Caco-2 to verify the effect of CDC40 on cell proliferation.The experimental data were statistically analyzed using IBM SPSS Statistics Version 22 software. Results In 70 patients,the CDC40 high-expression group accounted for 51.4 %.The expression level of CDC40 was related to the tumor differentiation degree,T stage,lymph node metastasis and the levels of Ki67,but not to gender,age and distant metastasis.Multivariate and univariate survival analyses showed that the high expression of CDC40 was associated with poorer survival prognosis of patients.In vitro experiments showed that overexpression of CDC40 promoted the proliferation of colorectal cancer cells.The nude mouse tumorigenesis experiment demonstrated that the overexpression of CDC40 promoted the tumor growth. Conclusion CDC40 plays an important role in the occurrence and development of colorectal cancer and can be used as a potential molecular marker and therapeutic target.

Objective To comprehensively analyze the expression,clinical significance,potential biological functions,and preliminary mechanisms of PAK6 in breast cancer,and to evaluate its feasibility as a prognostic biomarker and therapeutic target for breast cancer. Methods Breast cancer expression profile data from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)were utilized to analyze the expression of PAK6 and its relationship with patient prognosis.Potential biological functions of PAK6 were explored based on transcriptome data and single-cell RNA sequencing data.The overexpression and knockdown models of PAK6 were established in human breast cancer cell lines MCF7 and MDA-MB-231.JC-1 staining,apoptosis detection,and Western blotting were used to analyze the effects of PAK6 on cell apoptosis and DNA damage response. Results PAK6 was significantly overexpressed in breast cancer tissues,and its high expression was negatively correlated with overall survival,progression-free survival,disease-specific survival,and relapse-free survival in patients(P<0.05).GSEA results showed that the PAK6 high-expression group was enriched in pathways related to DNA replication,cell cycle regulation,and DNA damage repair.Single-cell analysis revealed that PAK6 was predominantly highly expressed in Malignant_C25,Malignant_C28,and Malignant_C3 malignant tumor cell subpopulations.Notably,the Malignant_C25 subpopulation showed enrichment for cell cycle and DNA replication-related pathways and exhibited the potential to transition to other cell types.In vitro experiments confirmed that PAK6 overexpression inhibited apoptosis and decreased the phosphorylation level of the DNA damage marker γH2AX in breast cancer cells,whereas PAK6 knockdown promoted apoptosis and increased γH2AX phosphorylation. Conclusion PAK6 is overexpressed in breast cancer and is associated with poor prognosis.It may play a pro-oncogenic role by promoting DNA replication,accelerating cell cycle progression,inhibiting apoptosis,and regulating DNA damage response.PAK6 shows promise as a potential prognostic biomarker and therapeutic target for breast cancer.

Objective To explore the effect of high mobility group box 1(HMGB1)on pathogenesis of vitiligo through inhibition of glycoprotein 100(gp100) and Nrf-2/HO-1/GPX4 expression,promotion of ferroptosis,and amplification of the inflammatory cascade. Methods A total of 24 C57BL/6 male mice aged 5-6 weeks were randomly divided into 4 groups,with 6 mice in each group:control group,model group,model +HMGB1 treatment group and model +gp100 degrading peptide treatment group.A mouse model of hydroquinone-induced vitiligo was established.The expression levels of HMGB1,gp100,the ferroptosis-related factors(GPX4,SLC7A11)and the endoplasmic reticulum stress-related factors(PERK,p-PERK,ATF6,GRP78,Nrf-2,HO-1)were detected by Western blotting.ELISA was used to measure the levels of TNF-α,IL-6 and MDA.DHE fluorescent probe was used to determine the ROS levels in mice skin tissues.Hematoxylin-eosin(HE)staining and histopathological analysis were performed in mice skin tissues. Results Compared with those in the control group,the score of depigmentation area,the levels of ROS,TNF-α,IL-6,MDA,and the expression levels of HMGB1,SLC7A11,p-PERK,ATF6 and GRP78 in the model group were increased(all P<0.01),whereas the expression levels of gp100 and GPX4 were decreased(P<0.01).The epidermal cells in the model group were disorderly arranged.In the HMGB1 treatment group,the expression levels of Nrf-2,HO-1 and gp100 were decreased(P<0.05),whereas the expression levels of p-PERK,ATF6 and GRP78 were increased(P<0.05).The levels of ROS,MDA and the expression level of SLC7A11 were increased in the gp100 degrading peptide treatment group(P<0.05),and the epidermal cells were more loosely arranged,with a large number of melanocytes undergoing apoptosis. Conclusion HMGB1 may be involved in inhibiting gp100 expression in melanocytes by mediating endoplasmic reticulum stress and inflammatory cascades,and down-regulating Nrf-2/HO-1/GPX4 to promote ferroptosis in melanocytes.

Objective To investigate the antibacterial activity of alginate-silver-S-nitrosoglutathione nanoparticle(NP) gel in the treatment of Helicobacter pylori(H.pylori)infected mouse model. Methods Alginate NPs gel,alginate-Ag NPs gel and alginate-Ag-GSNO NPs gel were prepared.The surface characteristics of Ag-GSNO nanoparticle gel were imaged by scanning electron microscopy.A mouse model of H.pylori infection was established.A total of 40 male C57BL/6 mice were randomly divided into 4 groups with 10 mice each:Vehicle group,Model group,Alginate-Ag NPs gel group,Alginate-AG-GSNO NPs gel group.The colony forming unit(CFU)counting of H.pylori infection in mice gastric epithelium was performed with blood agar culture.HE staining and histopathology were used to analyze the inflammation and injury of gastric epithelium in mice.The expression levels of ureB gene,which is a positive marker of H.pylori infection,was determined by qPCR.The levels of inflammatory cytokines TNF-α,IL-1β and IL-6 were determined by ELISA.The expression levels of NLRP3 inflammasome was determined by Westren blotting. Results Alginate-Ag nanoparticles had a spherical morphology,with a particle size range of 100-200 nm.Spray dried GSNO particles were spherical with slight depressions,and their size was at the μm level.Compared with those in the Alginate-Ag NPs gel group,the CFU of H.pylori infected mice gastric epithelial tissues was decreased(P<0.05),and the HE staining score was reduced(P<0.05)in the Alginate-AG-GSNO NPs gel group.The Gastric epithelial cells in the Alginate-AG-GSNO NPs gel group were arranged orderly in the Alginate-AG-GSNO NPs gel group,and there were a reduced number of polymorphonuclear infiltration,lymphoid aggregation and H.pylori infection positive cells,indicating the improvement of inflammation and injury of gastric epithelial tissues.The expression levels of ureB gene,the relative expression levels of NLRP3 inflammasome,and the levels of inflammatory cytokines TNF-α,IL-1β and IL-6 were all decreased(all P<0.05). Conclusion Compared with alginate-Ag NPs gel,alginate-AG-GSNO NPs gel could more effectively inhibit the colonization of H.pylori in infected mouse model and improve gastric epithelium damage.

Objective To investigate the effect of long non-coding RNA(lncRNA)CRNDE on macrophage pyroptosis and M1 polarization in ulcerative colitis(UC)mice and the potential mechanism. Methods The UC mouse model was established,and colon length,disease activity index(DAI)were measured to validate the model.RAW264.7 cells were divided into 4 groups:Ctrl group,pcDNA-null group,pcDNA-CRNDE group,and pcDNA-CRNDE+LY3214996 group.The expression levels of lncRNA CRNDE,cleaved-Caspase-1,cleaved-GSDMD,ERK1/2,and phosphorylated ERK1/2(p-ERK1/2)were detected by qRT-PCR or Western blotting.The proportion of F4/80⁺CD86⁺macrophages was measured by flow cytometry. Results In the model group,the DAI score was increased,the colon length was shortened,and the expression levels of lncRNA CRNDE,cleaved-Caspase-1,cleaved-GSDMD,and p-ERK1/2 were upregulated,and the proportion of F4/80⁺CD86⁺ macrophages was increased(all P<0.05).In RAW264.7 cells,lncRNA CRNDE and cleaved-Caspase-1、cleaved-GSDMD、p-ERK1/2 expression in the pcDNA-CRNDE group was upregulated when compared with that in the pcDNA-null group,and that was downregulated after LY3214996 treatment(all P<0.05). Conclusion lncRNA CRNDE promotes macrophage pyroptosis and M1 polarization in UC mice by activating the ERK1/2 pathway,suggesting it may be a potential therapeutic target for UC.

Objective To analyze the inhibitory effect of hyperoside on inhibiting the proliferation of bladder urothelial carcinoma(BLCA)cells and the mechanism. Methods Cell proliferation was detected by CCK-8 and colony formation assays.The expression level of nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)in cells under different concentration of hyperoside and intervention time,and the expression levels of mTOR,AKT,S6 cells and NOX4 proteins were detected by Western blotting. Results The proliferation ability of cells,number of cell colonies,volume and mass of transplanted tumor tissues,and expression level of Ki67 protein were significantly decreased in the hyperoside group(P<0.05).The expression level of NOX4 was significantly decreased with the increase of the treatment time and concentration of hyperoside(P<0.05).Compared with those in the control group,the expression levels of p-mTOR,p-S6,p-Akt and NOX4,and number of colonies were significantly decreased in the hyperoside group,while the above indexes were significantly increased in the NOX4 overexpression group and the hyperoside +NOX4overexpression group(P<0.05). Conclusion Hyperoside can inhibit BLCA proliferation by supressing the expression of NOX4.

Objective To explore the effect of erythropoietin-producing hepatocyte receptor B4(EPHB4)on the malignant progression of osteosarcoma and its mechanism. Methods The expression level of EPHB4 in U2OS,SaOS2-LM7,SaOS2,and 143B was detected by Western blotting.SaOS2-LM7 cells was divided into 4 groups:control group,si-NC group,si-EPHB4 group,and sEPHB4(soluble EPHB4 blocking antibody)group.Cell proliferation was detected by CCK-8 assay and EdU staining,cell apoptosis was detected by flow cytometry,cell invasion and migration were detected by Transwell assay and Wound healing assay.A SaOS2-LM7 cell line labeled with red fluorescent protein(RFP)which was stably silenced of EPHB4 was constructed(EPHB4-si-RNA-RFP).Twenty-four tumor-bearing nude mice were divided into 2 groups:the control-RFP group and the EPHB4-si-RNA-RFP group.The lung metastasis was detected 1-4 weeks later by using small animal in vivo imaging system.Immunohistochemistry was used to analyze the expression of EPHB4,cluster of differentiation 33(CD33),CYCLIN D1,matrix metalloproteinase 9(MMP-9),MMP-14,and β-CATENIN in tumor tissues 4 weeks later. Results The expression level of EPHB4 in the SaOS2-LM7,SaOS2,and 143B was significantly higher than that in the U2OS(all P<0.05).Compared with those in the control group,the expression level of EPHB4,the proliferation activity,and the EdU positive rate of SaOS2-LM7 in the sEPHB4 group and the si-EPHB4 group were significantly decreased,the apoptosis rate was significantly increased,and the migration rate and invasion rate were significantly decreased(all P<0.05).In addition,the relative expression levels of β-CATENIN,CYCLIN D1,MMP-9,and MMP-14 in the sEPHB4 group and the si-EPHB4 group were also significantly decreased when compared with those in the control group(all P<0.05).The lung metastasis at 3 weeks and 4 weeks was less in the EPHB4-siRNA-RFP group than in the control-RFP group(both P<0.05).The protein expression of EPHB4,CD33,CYCLIN D1,MMP-9,MMP-14,and β-CATENIN in the tumor tissues of the EPHB4-siRNA-RFP group was down-regulated(all P<0.05). Conclusion Silencing EPHB4 significantly inhibits the activation of the WNT/β-CATENIN signaling pathway in osteosarcoma cells,suppresses cell proliferation,migration,and invasion,and promotes cell apoptosis.

Objective To analyze and verify the mechanism of sophocarpine in inhibiting osteosarcoma cells through network pharmacology and in vivo and in vitro experiments. Methods The targets of sophocarpine were predicted by network pharmacology.The relationship between 5-hydroxytryptamine receptor 3A(HTR3A)and prognosis of osteosarcoma was analyzed by bioinformatics.HTR3A was knocked down in osteosarcoma cell lines U2OS and MG63,and its effects on cell proliferation,colony formation,apoptosis,and the expression levles of cell cycle-related proteins were detected.The inhibitory effects of sophocarpine on the growth of osteosarcoma cells and transplanted tumors were observed by in vitro and in vivo experiments. Results HTR3A was predicted as a key target of sophocarpine,and its high expression level was associated with poor prognosis in patients with osteosarcoma(log-rank P<0.05,P<0.05).HTR3A was highly expressed in osteosarcoma cells,and knockdown of HTR3A inhibited cell proliferation and colony formation,and induced cell cycle arrest and apoptosis(P<0.05).Sophocarpine effectively inhibited the growth of osteosarcoma in vitro and in vivo(P<0.05),and the HTR3A protein expression level was decreased after sophocarpine treatment in a dose-dependent manner(P<0.05).Moreover,overexpression of HTR3A reversed the inhibitory effect of sophocarpine on cell proliferation(P<0.05). Conclusion Sophocarpine can inhibit the proliferation and colony formation and promote apoptosis of osteosarcoma cells by inhibiting the expression of HTR3A.HTR3A is a potential key target for sophocarpine to exert anti-tumor effect in osteosarcoma.

Objective To investigate the effect of nucleolar protein 14(NOP14)overexpressi on proliferation and migration of ovarian cancer cells SKOV3 and the underlying mechanism. Methods The expression level of NOP14 in ovarian cancer cell lines(SKOV3,A2780,HO-8910,OVCAR)was detected by RT-qPCR and Western blotting.SKOV3 cells were transfected with the pcDNA-NOP14 plasmid.Cell proliferation was detected by colony formation assay and 5-Ethynyl-2′-deoxyuridine(EdU)staining.Tumor stem cell characteristics were analyzed via serum-free sphere formation assay.The proportion of CD133-positive(CD133⁺)cells was measured by flow cytometry.The epithelial-mesenchymal transition(EMT)process was evaluated by observing cell morphological change and detecting the EMT marker proteins(E-cadherin,N-cadherin,Vimentin).The expression level of nuclear receptor interacting protein 1(NRIP1),and Wnt/β-catenin pathway proteins were detected by RT-qPCR and Western blotting. Results The expression level of NOP14 was significantly lower in the ovarian cancer cell lines than that in the normal ovarian epithelial cells.Overexpression of NOP14 significantly inhibited the proliferation,stem cell characteristics,and EMT of SKOV3 cells(mesenchymal markers N-cadherin and Vimentin were downregulated,while the epithelial marker E-cadherin was upregulated).Moreover,Overexpression of NOP14 suppressed the expression of NRIP1,Wnt and β-catenin. Conclusion NOP14 can inhibit the proliferation,stem cell characteristics,and EMT process of ovarian cancer SKOV3 cells,and negatively regulate the expression of NRIP1,Wnt,β-catenin.

Candida albicans is a common opportunistic pathogen fungus in humans.The invasive infection it causes have a high mortality rate,posing significant challenges for clinical prevention and treatment.The graving problem of drug resistance is one of the primary causes for treatment failure,making it urgent to elucidate its drug resistance mechanisms and identify new drug targets.This article reviews the bidirectional regulatory mechanism of the MAPK signaling pathway in Candida albicans drug tolerance and host interaction:On one hand,it reveals how the MAPK signaling pathway mediates interactions between Candida albicans and its host,such as through immune evasion and inflammatory responses;On the other hand,it elucidates the MAPK-mediated resistance mechanisms in Candida albicans,including cell wall synthesis and remodeling,hyphal formation,oxidative stress,and biofilm formation,along with recent advances in drug research.This aims to provide new insights into understanding the pathogenicity and resistance of Candida albicans and to offer theoretical support for developing novel antifungal strategies targeting the MAPK pathway.

tRNA-derived small RNAs(tsRNAs)are generated from the precursor tRNA or the mature tRNA through specific enzymatic cleavage reactions and exhibit a wide range of biological functions.In the field of gynecology,tsRNAs have been demonstrated to be differentially expressed in various gynecological diseases,including ovarian cancer,endometrial cancer and endometriosis,and are closely associated with disease progression and prognosis.This review aims to summarize the progresses of current researchs on tsRNAs in gynecological disorders,with the purpose of providing new insights into their molecular mechanisms and potential applications in early diagnosis and precision medicine.

Subarachnoid hemorrhage(SAH)is a critical neurological emergency.The mechanisms of brain injury following SAH are complex and multifaceted,with neuronal death serving as a pivotal pathological process.This article reviews recent researches on neuronal death mechanisms following SAH,covering apoptosis,pyroptosis,autophagy,necroptosis,ferroptosis,and other pathways.It also preliminarily explores relevant intervention strategies,aiming to provide new insights for the clinical management of SAH.

The intestinal barrier constitutes a critical defense system that separates the human body from the external environment.Its functional impairment is a hallmark of the aging process.As individuals age,the structure and function of the intestinal barrier gradually deteriorate,manifesting as increased intestinal permeability,impaired immune function,and dysbiosis of the gut microbiota,involving complex alterations at multiple physiological,cellular,and molecular levels.Such impairments not only compromise local intestinal homeostasis but may also induce systemic inflammation and immune dysfunction,thereby contributing to the overall acceleration of aging.This article reviews the characteristics,mechanisms,and systemic health implications of intestinal barrier aging,and explores potential intervention strategies.

Large language model(LLM)offers new opportunities for medical education.This paper examines its technical foundations and applications in case generation,personalized teaching,and intelligent assessment.By selecting appropriate models and prompts,and incorporating multimodal integration and academic validation,LLM can generate simulated teaching cases for clinical reasoning training.By combining student profiles and learning behaviors,LLM enables path recommendations and content customization,enhancing teaching adaptability.LLM also helps build multidimensional scoring systems focusing on medical accuracy,logical structure,and causal reasoning,providing personalized feedback.Despite challenges like hallucinations and outdated knowledge,future improvements should focus on human-machine collaboration,data security,and standardized resources to promote intelligent and standardized medical education.