Background: Malignant pleural effusion (MPE), a common complication of advanced cancers, is associated with poor prognosis and reduced quality of life. Although host–tumor interactions are known to drive MPE development, the associated immune dynamics during disease progression remain unclear. Using a Lewis lung carcinoma-induced MPE model in C57BL/6JNidfc mice, we systematically evaluated general parameters and immune cell changes at two-day intervals throughout disease progression. Results: The day of Lewis lung carcinoma cell injection into the pleural space was designated as day 0. By day 10 post-injection (p.i.), MPE-bearing mice exhibited ~ 10% body weight loss, marking the experimental endpoint. Pleural tumor mass and pleural effusion volume were minimal up to day 4 p.i. but increased sharply from day 6 onward.CD45⁺ immune cell counts rose over time, and days 6, 8, and 10 p.i. marked key stages of MPE progression. On day 6, B cells, T cells, and natural killer cells, but not macrophages and neutrophils, increased significantly compared to earlier timepoints. By day 8, all immune cell subsets except T cells exceeded day 6 levels, and at day 10, natural killer cell numbers declined while others continued to increase. Besides, the numbers of CD8⁺ T cells, Th1 cells, regulatory T cells, and M2 macrophages progressively increased from day 6 to 10. Based on these data, days 6 and 10 were defined as early and advanced MPE stages, respectively, with distinct immune phenotypes. In advanced MPE, CD8⁺ T cells displayed reduced IFN-γ, TNF-α, Granzyme B, Perforin, FasL, and Ki-67, but upregulated PD-1 and CTLA-4 relative to early stage. Similarly, Th1 cells showed decreased IFN-γ, TNF-α, and IL-2 production along with reduced Ki-67 expression. Advanced-stage M2 macrophages exhibited lower MHC-II levels and impaired phagocytosis, but higher PD-L1 and IL-10 production, while neutrophils showed reduced TNF-α release and phagocytic activity. Conclusions Our findings characterize the temporal immune dynamics associated with MPE progression in a mouse model, revealing a transition from an early immunostimulatory state to a late immunosuppressive state. This study enhances our understanding of MPE immunopathogenesis and provides a foundation for developing precise, stagespecific therapeutic strategies.

This study was aimed at determining the genetic characteristics of Yersinia pestis in Subei County through differ-ential region(DFR),clustered regularly interspaced short palindromic repeats(CRISPR),and variable number tandem repeat(VNTR)analyses,to guide the tracing of plague outbreaks.The DNA of 89 Yersinia pestis strains isolated from various ani-mal in foci of Subei County from 1973 to 2017 was extracted.Primers for genotyping by DFR,CRISPR,and MLVA were used in PCR,and agarose electrophoresis was used to determine whether the amplified products were present.Genotypes were deter-mined through comparison against the DFR database of Yersinia pestis in China.The PCR products were sequenced and com-pared against the online CRISPR database for Yersinia pestis to determine the genotype.The number of VNTR repeats in each strain was calculated through capillary electrophoresis,and the minimum spanning tree was constructed with BioNumerics 7.6 according to the numbers of VNTR repeats from 89 Yersinia pestis strains in Subei County and 11 strains from a Marmota hi-malayana focus in Qinghai Province.The Yersinia pestis strains in Subei County were divided into six main genotypes by DFR:8,7,lb,5,32,and 44.The Yersinia pestis strains were divided into three gene clusters and three genotypes by CRISPR.Ca35'was the main gene cluster in Subei County;the genotype was 26';and the distribution was primarily in Dangchengwan Town,Yuerhong Township,and Shibaocheng Township.Ca7 and CaΔ5'comprised secondary gene clusters,with genotypes 22 and 24,and were distributed in Dangchengwan Town,Yuerhong Township,and Shibaocheng Township.The Yersinia pes-tis strains in Subei County were divided primarily into three clusters:the Dangchengwan Machang cluster,Yuerhong Township cluster and Dangchengwan Town cluster.Therefore,the Yersinia pestis strains in Subei County,divided into major and minor genotypes according to DFR,CRISPR and MLVA,showed different regional distribution characteristics,highly diverse geno-types,and complex population characteristics.These aspects are particularly important in tracing the sources of plague out-breaks.

Objective To evaluate the efficacy and safety of a novel intravascular lithotripsy(IVL)balloon—Vesscrack shockwave balloon—for vascular preparation before stent implantation in patients with severe coronary artery calcification(CAC).Methods This was a prospective,single-arm,multicenter study conducted in China from June 2022 to October 2022.Patients with severe CAC were treated with the Vesscrack shockwave balloon for lesion preparation,followed by drug-eluting stent(DES)implantation.Of these,33 patients underwent optical coherence tomography(OCT).The primary endpoint was procedural success,defined as successful stent implantation with residual stenosis≤30％and the absence of in-hospital major adverse events,including cardiac death,target vessel-related myocardial infarction,or target lesion revascularization.Results A total of 170 patients[mean age:(65.9±7.9)years,116 males]were enrolled.After treatment with IVL and DES,the minimum lumen diameter increased significantly compared to baseline[(2.34±0.40)mm vs.(0.95±0.33)mm,P＜0.001],the degree of stenosis was significantly reduced[(13.24±6.60)％vs.(65.18±10.59)％,P＜0.001].Procedural success was achieved in 100％of cases,and device success was 98.8％.The 30-day patient-related cardiovascular clinical composite endpoint(POCE)rate was 0.0,with no target lesion failure,no confirmed or potential thrombotic events were observed.The shockwave energy generator demonstrated excellent stability and ease of use.Among the 33 patients assessed with OCT,after IVL intervention,the maximum calcified area of the lumen[(3.51±1.51)mm2 vs.(2.85±1.80)mm2,P＜0.001],and the minimum lumen area within the target lesion[(3.08±1.04)mm2 vs.(2.02±0.75)mm2,P＜0.001],and after DES intervention,the luminal area of the largest calcified site[(6.59±1.64)mm2 vs.(2.85±1.80)mm2,P＜0.001]and the minimum luminal area within the target lesion[(6.19±1.45)mm2 vs.(2.02±0.75)mm2,P＜0.001]were significantly increased,and the differences were statistically significant.Conclusions The Vesscrack shockwave balloon is effective and safe for vascular preparation in patients with severe CAC prior to stent implantation.It achieves significant calcified plaque modification,high procedural success rates,and minimal complications.

This study systematically investigated the molecular mechanisms underlying the involvement of mesoderm development-associated genes in melanoma progression through integrated bioinformatics analy-sis and experimental validation.Utilizing the GSVA(gene set variation analysis)algorithm to perform enrichment analysis of 7 752 biological functions in 406 skin cutaneous melanoma(SKCM)cases,we i-dentified for the first time the significant activation of mesoderm development pathways during SKCM pathogenesis.Four core regulatory genes(SMAD4,NODAL,BMPR1A,and ZFP36L1)were screened using LASSO-COX regression analysis and a prognostic risk-scoring system was established.Gene Ontolo-gy(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses revealed predomi-nant enrichment of these genes in mRNA metabolic processes and TGF-β signaling pathways.Experimen-tal validation through Quantitative Polymerase Chain Reaction(qPCR),Western blotting,and immuno-histochemistry(IHC)demonstrated that:(1)Downregulation of SMAD4 and BMPR1A in tumor tissues was significantly correlated with poor prognosis(P＜0.05);(2)NODAL promoted tumor invasion and metastasis by regulating epithelial-mesenchymal transition(EMT);(3)High ZFP36L1 expression was associated with enhanced chemotherapy sensitivity.Further analyses revealed significant correlations be-tween core gene expression levels and tumor immune infiltration characteristics as well as immune check-point molecules.By integrating multi-omics analysis with experimental validation,this study elucidates the critical roles of mesoderm development-associated genes in SKCM progression,particularly clarifying the molecular mechanisms through which SMAD4/NODAL/BMPR1A/ZFP36L1 influence tumor biologi-cal behaviors via immune microenvironment regulation and EMT processes.These findings provide novel theoretical foundations for molecular subtyping and targeted therapy in melanoma.

Objective:To analyze intestinal microecological differences and their correlation with psychological status in patients with constipated irritable bowel syndrome(IBS-C)and functional constipation(FC).Methods:A total of 160 patients with IBS-C and FC attending the outpatient and inpatient departments of the Department of Gastroenterology at the General Hospital of Ningxia Medical University from January 2021 to January 2024 were selected and grouped according to the type of disease into the IBS-C group(80 patients)and the FC group(80 patients),and 80 patients who had a health checkup during the same period were selected as the control group.Fresh fecal specimens were taken from all subjects,and 16S rRNA sequencing was used to detect the intestinal flora,and the indices of intestinal microecological diversity(Ace index,Chao index,Shannon index,and Sobs index)were detected,and the psychological state of the subjects was assessed by using the Self-Assessment Scale for Anxiety(SAS)and the Self-Depression Scale(SDS).Pearson's correlation method was used to analyze the correlation between intestinal microecology and correlation with psychological state in IBS-C and FC patients.Result:Bifidobacteria and Lactobacillus levels were lower in both the FC and IBS-C groups compared with the control group,and the decrease was more pronounced in the IBS-C group(P＜0.05).Enterobacteriaceae and Enterococci contents were higher in patients in the FC and IBS-C groups(P＜0.05),with the greatest increase in the IBS-C group(P＜0.05);Microbial diversity analysis showed that there were differences in Ace,Chao,Shannon and Sobs indices among the three groups(P＜0.05).All diversity indices were lower in the FC and IBS-C groups than in the control group,and the degree of reduction was more pronounced in the IBS-C group than in the FC group(P＜0.05);The SAS and SDS scores of both FC and IBS-C groups were higher than those of the control group,and the scores of the IBS-C group were higher than those of the FC group(P＜0.05);correlation analysis showed that,in the IBS-C and FC groups,Bifidobacterium and Lactobacillus,Ace index,Chao index,Shannon index,and Sobs index were negatively correlated with SAS and SDS scores.Enterobacteriaceae and Enterococci were positively correlated with SAS and SDS scores(P＜0.05).Conclusion:There are possible differences in the gut microbiota between IBS-C and FC patients.The imbalance of gut microbiota is more significant in IBS-C patients,and gut microbiota is closely related to psychological status.Clinical treatment of IBS-C and FC should pay attention to the regulation of gut microbiota and intervention of psychological status.

This study aims to investigate the effects of aflatoxin B1(AFB1)on the infection and replication of porcine delta coronavirus(PDCoV).Porcine small intestinal epithelial cells(IPEC-J2),porcine kidney cells(LLC-PK)and swine testis cells(ST)were used as models,and CCK-8 method was used to detect the safe mass concentration range of AFB1 on the three cell lines.Each cell line was divided into the blank control group,AFB1 treatment group,PDCoV treatment group and AFB1 and the PDCoV co-treatment group.The cells were treated with safe mass concentration of AFB1 for 12 h,and then treated with PDCoV for 20 h.Total RNA and total protein were extrac-ted from the cells.The effects of AFB1 on PDCoV infection and replication were detected by qPCR,Western blot and cell immunofluorescence tests.The results showed that compared with the PDCoV treatment group,the mRNA and N protein of viral S protein in the AFB1 and PDCoV co-treatment group were significantly increased(P＜0.05),and a significantly higher fluorescence of PDCoV N protein is also visibly present in the co-treatment group,and there was a significant difference between the two groups.The results showed that AFB1 could promote the infection and replication of PDCoV.It provides important data that AFB1 can promote the infection replication of PDCoV and provides a new idea for the prevention and treatment of PDCoV.

This study systematically investigated the molecular mechanisms underlying the involvement of mesoderm development-associated genes in melanoma progression through integrated bioinformatics analy-sis and experimental validation.Utilizing the GSVA(gene set variation analysis)algorithm to perform enrichment analysis of 7 752 biological functions in 406 skin cutaneous melanoma(SKCM)cases,we i-dentified for the first time the significant activation of mesoderm development pathways during SKCM pathogenesis.Four core regulatory genes(SMAD4,NODAL,BMPR1A,and ZFP36L1)were screened using LASSO-COX regression analysis and a prognostic risk-scoring system was established.Gene Ontolo-gy(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses revealed predomi-nant enrichment of these genes in mRNA metabolic processes and TGF-β signaling pathways.Experimen-tal validation through Quantitative Polymerase Chain Reaction(qPCR),Western blotting,and immuno-histochemistry(IHC)demonstrated that:(1)Downregulation of SMAD4 and BMPR1A in tumor tissues was significantly correlated with poor prognosis(P＜0.05);(2)NODAL promoted tumor invasion and metastasis by regulating epithelial-mesenchymal transition(EMT);(3)High ZFP36L1 expression was associated with enhanced chemotherapy sensitivity.Further analyses revealed significant correlations be-tween core gene expression levels and tumor immune infiltration characteristics as well as immune check-point molecules.By integrating multi-omics analysis with experimental validation,this study elucidates the critical roles of mesoderm development-associated genes in SKCM progression,particularly clarifying the molecular mechanisms through which SMAD4/NODAL/BMPR1A/ZFP36L1 influence tumor biologi-cal behaviors via immune microenvironment regulation and EMT processes.These findings provide novel theoretical foundations for molecular subtyping and targeted therapy in melanoma.

Objective:To analyze intestinal microecological differences and their correlation with psychological status in patients with constipated irritable bowel syndrome(IBS-C)and functional constipation(FC).Methods:A total of 160 patients with IBS-C and FC attending the outpatient and inpatient departments of the Department of Gastroenterology at the General Hospital of Ningxia Medical University from January 2021 to January 2024 were selected and grouped according to the type of disease into the IBS-C group(80 patients)and the FC group(80 patients),and 80 patients who had a health checkup during the same period were selected as the control group.Fresh fecal specimens were taken from all subjects,and 16S rRNA sequencing was used to detect the intestinal flora,and the indices of intestinal microecological diversity(Ace index,Chao index,Shannon index,and Sobs index)were detected,and the psychological state of the subjects was assessed by using the Self-Assessment Scale for Anxiety(SAS)and the Self-Depression Scale(SDS).Pearson's correlation method was used to analyze the correlation between intestinal microecology and correlation with psychological state in IBS-C and FC patients.Result:Bifidobacteria and Lactobacillus levels were lower in both the FC and IBS-C groups compared with the control group,and the decrease was more pronounced in the IBS-C group(P＜0.05).Enterobacteriaceae and Enterococci contents were higher in patients in the FC and IBS-C groups(P＜0.05),with the greatest increase in the IBS-C group(P＜0.05);Microbial diversity analysis showed that there were differences in Ace,Chao,Shannon and Sobs indices among the three groups(P＜0.05).All diversity indices were lower in the FC and IBS-C groups than in the control group,and the degree of reduction was more pronounced in the IBS-C group than in the FC group(P＜0.05);The SAS and SDS scores of both FC and IBS-C groups were higher than those of the control group,and the scores of the IBS-C group were higher than those of the FC group(P＜0.05);correlation analysis showed that,in the IBS-C and FC groups,Bifidobacterium and Lactobacillus,Ace index,Chao index,Shannon index,and Sobs index were negatively correlated with SAS and SDS scores.Enterobacteriaceae and Enterococci were positively correlated with SAS and SDS scores(P＜0.05).Conclusion:There are possible differences in the gut microbiota between IBS-C and FC patients.The imbalance of gut microbiota is more significant in IBS-C patients,and gut microbiota is closely related to psychological status.Clinical treatment of IBS-C and FC should pay attention to the regulation of gut microbiota and intervention of psychological status.

Objective To evaluate the efficacy and safety of a novel intravascular lithotripsy(IVL)balloon—Vesscrack shockwave balloon—for vascular preparation before stent implantation in patients with severe coronary artery calcification(CAC).Methods This was a prospective,single-arm,multicenter study conducted in China from June 2022 to October 2022.Patients with severe CAC were treated with the Vesscrack shockwave balloon for lesion preparation,followed by drug-eluting stent(DES)implantation.Of these,33 patients underwent optical coherence tomography(OCT).The primary endpoint was procedural success,defined as successful stent implantation with residual stenosis≤30％and the absence of in-hospital major adverse events,including cardiac death,target vessel-related myocardial infarction,or target lesion revascularization.Results A total of 170 patients[mean age:(65.9±7.9)years,116 males]were enrolled.After treatment with IVL and DES,the minimum lumen diameter increased significantly compared to baseline[(2.34±0.40)mm vs.(0.95±0.33)mm,P＜0.001],the degree of stenosis was significantly reduced[(13.24±6.60)％vs.(65.18±10.59)％,P＜0.001].Procedural success was achieved in 100％of cases,and device success was 98.8％.The 30-day patient-related cardiovascular clinical composite endpoint(POCE)rate was 0.0,with no target lesion failure,no confirmed or potential thrombotic events were observed.The shockwave energy generator demonstrated excellent stability and ease of use.Among the 33 patients assessed with OCT,after IVL intervention,the maximum calcified area of the lumen[(3.51±1.51)mm2 vs.(2.85±1.80)mm2,P＜0.001],and the minimum lumen area within the target lesion[(3.08±1.04)mm2 vs.(2.02±0.75)mm2,P＜0.001],and after DES intervention,the luminal area of the largest calcified site[(6.59±1.64)mm2 vs.(2.85±1.80)mm2,P＜0.001]and the minimum luminal area within the target lesion[(6.19±1.45)mm2 vs.(2.02±0.75)mm2,P＜0.001]were significantly increased,and the differences were statistically significant.Conclusions The Vesscrack shockwave balloon is effective and safe for vascular preparation in patients with severe CAC prior to stent implantation.It achieves significant calcified plaque modification,high procedural success rates,and minimal complications.

Objective To establish a backpack-based field emergency medical rescue equipment system to enhance emergency rescue efficiency.Methods A backpack-based field emergency medical rescue equipment system was preliminarily constructed with the concept of medical treatment in echelons and prolonged field care(PFC)and the method of capability-based equipment need analysis;with the Delphi method 15 experts from relevant fields were invited to execute two rounds of questionnaire consultations,and the final equipment system was determined after the equipment varieties and quantities were revised based on the expert opinions.Results The response rate for the two rounds of expert questionnaire surveys was 100%.The experts'authority coefficients were 0.8734 and 0.87,respectively;Kendall coefficients were 0.232 and 0.345(both P<0.001),indicating statistically significant results.Ultimately,a backpack-based field emer-gency medical rescue equipment system comprising 15 backpacks and 158 individual pieces of equipment was established.Conclusion The established system demonstrates a certain degree of specificity and practicability,providing references for the equipment allocation and utilization of emergency medical rescue teams.[Chinese Medical Equipment Journal,2025,46(10):17-22]