Objective To investigate the causal relationship between metabolic syndrome and breast cancer by using a bidirectional two-sample Mendelian randomization (MR) approach. Methods Genome-wide association study (GWAS) summary statistics for metabolic syndrome and breast cancer were acquired from the Integrative Epidemiology Unit GWAS database and the GWAS Catalog, with populations encompassing the United States and East Asia. A bidirectional causal design was employed: a forward analysis with metabolic syndrome as the exposure and breast cancer as the outcome, followed by a reverse analysis wherein their roles were interchanged. The inverse-variance weighting (IVW) method was primarily used for effect estimation, supplemented by MR-Egger regression, the weighted median method, the simple mode method, and the weighted mode method. Instrument variable strength was screened using the F-statistic (F>10). Robustness of the results was assessed through heterogeneity tests, horizontal pleiotropy tests, forest plots, and leave-one-out sensitivity analyses. Results The IVW analysis indicated no significant causal relationship between metabolic syndrome and breast cancer (OR=1.00, 95%CI: 0.97-1.03), P>0.05). Sensitivity analyses yielded consistent results, suggesting the good robustness of the study findings. Conclusion This study found no evidence to support a causal relationship, either positive or negative, between metabolic syndrome and breast cancer.

Objective To investigate the expression and clinical significance of 12 plasma cytokines in patients with pancreatic cancer.Methods The study included 120 patients with pancreatic cancer diagnosed and treated at Qingdao University Affiliated Hospital and 68 healthy controls from March 2023 to June 2024.The levels of 12 plasma cytokines,including interleukin(IL)-1 β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12P,IL-17,interferon(IFN)-α,IFN-γ,and tumor necrosis factor(TNF)-α,were detected using mul-tiplex bead-based flow immunoassay.The levels of tumor markers carcinoembryonic antigen(CEA),carbohydrate antigen 19-9(CA19-9)and carbohydrate antigen 72-4(CA 72-4)in the serum of pancreatic cancer patients were detected by electrochemilumines-cence,while carbohydrate antigen 242(CA242)were detected by chemiluminescence methods.The correlation between the expression levels of differentially expressed cytokines and those of tumor markers was analyzed using Spearman's rank correlation.Results The plasma levels of 9 cytokines(IL-1 β,IL-4,IL-5,IL-6,IL-8,IL-10,IL-17,IFN-α,and IFN-γ)in the patients with pancreatic canc-er were significantly higher than those in controls(all P＜0.05).The levels of IL-1 β,IL-6,IL-8,IL-10,and IL-17 in the advanced pancreatic cancer group were significantly higher than those in the early-stage group(P＜0.05).The plasma IL-6 level in the poorly dif-ferentiated pancreatic cancer group was significantly higher than that in the well-differentiated group(P＜0.05).The serum levels of CEA,CA19-9,CA242,and CA72-4 in the advanced pancreatic cancer group were significantly higher than those in the early-stage group(P＜0.05).The serum CEA level in the poorly differentiated pancreatic cancer group was significantly higher than that in the moderately differentiated pancreatic cancer group(P＜0.05).After four cycles of chemotherapy,IL-8 levels in the disease control group were significantly reduced compared to pre-treatment levels(P＜0.05),while IL-6,IL-8,and IL-10 levels in the disease progression group were significantly elevated compared to pre-treatment levels(all P＜0.05).In the patients with pancreatic cancer,plasma IL-6 levels were positively correlated with serum CEA levels(rs=0.238,P＜0.01)and serum CA19-9 levels(rs=0.186,P＜0.05).The plasma IL-10 levels were positively correlated with serum CA72-4(rs=0.220,P＜0.05)levels in the patients.Conclusion Nine cyto-kines in plasma,such as IL-6,etc.may be involved in the formation of the inflammatory microenvironment of pancreatic cancer,as well as in the proliferation and differentiation of tumor cells.The determination of their plasma levels should be helpful for the assessing disease conditions and therapeutic effects.

Objective:To study the etiological relationship between Yunnan sudden unexplained death (hereinafter referred to as YNSUD) and the desmosomal protein gene mutation of arrhythmogenic right ventricular cardiomyopathy (ARVC).Methods:From September 2019 to August 2020, a cross-sectional survey method was used to select 9 key counties (cities) of YNSUD in Yunnan Province as survey sites. Autopsy cardiac blood samples of YNSUD cases ( n = 11) were collected, and peripheral venous blood samples of co-occurring case ( n = 1), case relatives ( n = 128), and control population ( n = 60) were collected. Genomic DNA from blood was extracted. After PCR amplification, 97 exons of 5 ARVC desmosomal protein genes, including plakophilin-2 (PKP2), desmoglein-2 (DSG2), desmocollin-2 (DSC2), desmoplakin (DSP), and junction plakoglobin (JUP) were sequenced by Sanger method, and the gene mutation was analyzed. Results:Compared with the control population, YNSUD cases, co-occurring case and case relatives carried 52 gene mutation sites in 36 exons of the ARVC desmosomal protein gene, with a total mutation rate of 37.11% (36/97). Among them, there were 21 in DSP gene, 10 in DSG2 gene, 8 in PKP2 gene, 8 in DSC2 gene, and 5 in JUP gene. YNSUD cases, co-occurring case and case relatives carried two same gene mutation sites: DSG2 gene exon 15 c.3321 T>C synonymous mutation and JUP gene exon 3 c.213 T>C synonymous mutation.Conclusions:The mutation rate of ARVC desmosomal protein gene is relatively high in the population of YNSUD. The two same gene mutation sites (DSG2 gene c.3321 T>C and JUP gene c.213 T>C) carried by YNSUD cases, co-occurring case and case relatives may be associated with the pathogenesis of YNSUD.

Objective To investigate the expression and clinical significance of 12 plasma cytokines in patients with pancreatic cancer.Methods The study included 120 patients with pancreatic cancer diagnosed and treated at Qingdao University Affiliated Hospital and 68 healthy controls from March 2023 to June 2024.The levels of 12 plasma cytokines,including interleukin(IL)-1 β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12P,IL-17,interferon(IFN)-α,IFN-γ,and tumor necrosis factor(TNF)-α,were detected using mul-tiplex bead-based flow immunoassay.The levels of tumor markers carcinoembryonic antigen(CEA),carbohydrate antigen 19-9(CA19-9)and carbohydrate antigen 72-4(CA 72-4)in the serum of pancreatic cancer patients were detected by electrochemilumines-cence,while carbohydrate antigen 242(CA242)were detected by chemiluminescence methods.The correlation between the expression levels of differentially expressed cytokines and those of tumor markers was analyzed using Spearman's rank correlation.Results The plasma levels of 9 cytokines(IL-1 β,IL-4,IL-5,IL-6,IL-8,IL-10,IL-17,IFN-α,and IFN-γ)in the patients with pancreatic canc-er were significantly higher than those in controls(all P＜0.05).The levels of IL-1 β,IL-6,IL-8,IL-10,and IL-17 in the advanced pancreatic cancer group were significantly higher than those in the early-stage group(P＜0.05).The plasma IL-6 level in the poorly dif-ferentiated pancreatic cancer group was significantly higher than that in the well-differentiated group(P＜0.05).The serum levels of CEA,CA19-9,CA242,and CA72-4 in the advanced pancreatic cancer group were significantly higher than those in the early-stage group(P＜0.05).The serum CEA level in the poorly differentiated pancreatic cancer group was significantly higher than that in the moderately differentiated pancreatic cancer group(P＜0.05).After four cycles of chemotherapy,IL-8 levels in the disease control group were significantly reduced compared to pre-treatment levels(P＜0.05),while IL-6,IL-8,and IL-10 levels in the disease progression group were significantly elevated compared to pre-treatment levels(all P＜0.05).In the patients with pancreatic cancer,plasma IL-6 levels were positively correlated with serum CEA levels(rs=0.238,P＜0.01)and serum CA19-9 levels(rs=0.186,P＜0.05).The plasma IL-10 levels were positively correlated with serum CA72-4(rs=0.220,P＜0.05)levels in the patients.Conclusion Nine cyto-kines in plasma,such as IL-6,etc.may be involved in the formation of the inflammatory microenvironment of pancreatic cancer,as well as in the proliferation and differentiation of tumor cells.The determination of their plasma levels should be helpful for the assessing disease conditions and therapeutic effects.

Objective:To analyze the infection status of Coxsackievirus B group (CVB) in regions affected by sudden unexplained death in Yunnan (referred to as sudden death in Yunnan), and to provide a scientific basis for formulating effective prevention and control strategies.Methods:A cross-sectional survey method was employed. The population from 16 counties (cities, districts, referred to as counties) affected by sudden death in Yunnan Province from 2002 to 2022 and the population from one non-affected county in 2021 and 2022 (control population) were classified into cases of sudden death in Yunnan (7 cases), co-occurring cases (29 cases), high-risk population (1 303 cases), and control population (270 cases). Blood samples were collected from these populations. By using enzyme-linked immunosorbent assay (ELISA), CVB immunoglobulin M (CVB-IgM) antibodies in the acute-phase serum samples of the population in the affected areas were detected, and CVB immunoglobulin G (CVB-IgG) antibodies in the convalescent-phase serum samples were detected. Both types of detections were carried out on the control population, and the test results were analyzed.Results:A total of 1 609 serum samples were tested, including 1 339 samples from the population in the affected areas (923 acute-phase samples and 416 convalescent-phase samples) and 270 samples from the control population. Among the 16 affected counties, positive CVB-IgM antibody results were detected in 9 counties. The overall positive rate of the population in the affected areas was higher than that of the control population [7.80% (72/923) vs. 4.44% (12/270), χ 2 = 40.78, P < 0.001]. The positive rates of the high-risk population in Dayao County and Lufeng City were both higher than that of the control population [(22.22% (22/99), 10.92% (25/229) vs. 4.44% (12/270), χ 2 = 27.37, 7.56, P < 0.05]. Positive CVB-IgG antibody results were detected in 7 counties. The overall positive rate of the population in the affected areas was higher than that of the control population [(4.09% (17/416) vs. 0.74% (2/270), χ 2 = 6.81, P = 0.009]. The positive rates of CVB-IgM and CVB-IgG antibodies in the population of the affected areas in Dayao County [22.22% (22/99), 9.80% (5/51)] were both higher than those of the control population ( P < 0.05). Among the five affected villages in Dayao County, the positive rates of CVB-IgM and CVB-IgG antibodies in the population of Aji Ju Village were the highest [25.49% (13/51), 3/13]. Conclusions:The positive rates of both CVB-IgM and CVB-IgG antibodies in the population of the areas affected by sudden death in Yunnan were higher than those of the control population, indicating that CVB infection occurred during the sudden death events in the above-mentioned affected areas.

Objective To investigate the gene expression profile in rat pancreatic ductal stem cells(PDSCs)when induced to differentiate into insulin-secreting cells(IPCs),with the goal of identifying key genes involved in this differentiation process.Methods The expanded PDSCs were categorized into a normal control(NC)group and an induced(Tre)group.PDSCs continued expansion culture in NC group,and cultured in induction medium for 28 days to facilitate the differentiation of PDSCs into IPCs in Tre group.Dithizone staining was employed to morphologically assess whether the cells exhibited a reddish-brown coloration,indicating a positive result.The immunofluorescence staining method was used to detect the expression of insulin(Ins)and PDX1 in the cells following induction.Additionally,ELISA was conducted to measure the Ins release from IPCs,thereby verifying the responsiveness of the induced cells to glucose-stimulated Ins secretion.Concurrently,cells were collected on induction days 0 and 28 for RNA sequencing(RNA-seq),and differentially expressed genes(DEGs)were analyzed and functionally annotated.The analysis revealed that regulatory factor X3(RFX3)was overexpressed in PDSCs,and the impact of RFX3 upregulation on differentiation induction was subsequently verified.Results Compared with NC group,DTZ staining was positive,PDX1 and Ins proteins were expressed,and an increased release of Ins in response to sugar stimulation was demonstrated in the Tre group.RNA-seq analysis identified 4270 DEGs,and functional enrichment analysis utilizing the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed associations with Ins response,positive regulation of Ins secretion,pancreatic endocrine cell development,and overall pancreatic development.Additionally,functionally related genes such as ALDHA2,CREB5,EIF6,FOXO1,RFX3,WNT5a,OGT,GPR39,SMAD6,and TRPM2 were identified,indicating involvement in the cell cycle,TGF-β1 signaling pathway,FOXO signaling pathway,and Wnt signaling pathway in the regulation of the differentiation of pancreatic ductal stem cells(PDSCs)into insulin-producing cells(IPCs).Furthermore,the upregulation of RFX3 can inhibit the expression of TGF-β1 within 72 hours,thereby promoted the formation and release of Ins from insulin-positive cells.Conclusions Multiple genes and signaling pathways associated with pancreatic β-cell function collectively regulate the differentiation of rat PDSCs into IPCs.Notably,the upregulation of RFX3 enhances this differentiation process.

Objective:To analyze the infection status of Coxsackievirus B group (CVB) in regions affected by sudden unexplained death in Yunnan (referred to as sudden death in Yunnan), and to provide a scientific basis for formulating effective prevention and control strategies.Methods:A cross-sectional survey method was employed. The population from 16 counties (cities, districts, referred to as counties) affected by sudden death in Yunnan Province from 2002 to 2022 and the population from one non-affected county in 2021 and 2022 (control population) were classified into cases of sudden death in Yunnan (7 cases), co-occurring cases (29 cases), high-risk population (1 303 cases), and control population (270 cases). Blood samples were collected from these populations. By using enzyme-linked immunosorbent assay (ELISA), CVB immunoglobulin M (CVB-IgM) antibodies in the acute-phase serum samples of the population in the affected areas were detected, and CVB immunoglobulin G (CVB-IgG) antibodies in the convalescent-phase serum samples were detected. Both types of detections were carried out on the control population, and the test results were analyzed.Results:A total of 1 609 serum samples were tested, including 1 339 samples from the population in the affected areas (923 acute-phase samples and 416 convalescent-phase samples) and 270 samples from the control population. Among the 16 affected counties, positive CVB-IgM antibody results were detected in 9 counties. The overall positive rate of the population in the affected areas was higher than that of the control population [7.80% (72/923) vs. 4.44% (12/270), χ 2 = 40.78, P < 0.001]. The positive rates of the high-risk population in Dayao County and Lufeng City were both higher than that of the control population [(22.22% (22/99), 10.92% (25/229) vs. 4.44% (12/270), χ 2 = 27.37, 7.56, P < 0.05]. Positive CVB-IgG antibody results were detected in 7 counties. The overall positive rate of the population in the affected areas was higher than that of the control population [(4.09% (17/416) vs. 0.74% (2/270), χ 2 = 6.81, P = 0.009]. The positive rates of CVB-IgM and CVB-IgG antibodies in the population of the affected areas in Dayao County [22.22% (22/99), 9.80% (5/51)] were both higher than those of the control population ( P < 0.05). Among the five affected villages in Dayao County, the positive rates of CVB-IgM and CVB-IgG antibodies in the population of Aji Ju Village were the highest [25.49% (13/51), 3/13]. Conclusions:The positive rates of both CVB-IgM and CVB-IgG antibodies in the population of the areas affected by sudden death in Yunnan were higher than those of the control population, indicating that CVB infection occurred during the sudden death events in the above-mentioned affected areas.

AIM:A strain was isolated and identified as the H3N8 subtype of the avian influenza virus from a sick chicken at a farm in Yangjiang,Guangdong Province,named A/chicken/Yangjiang/552/2023(abbreviated as YJ/552).The aim of this research is to determine its genetic evolution,biological properties and pathogenicity in hamsters.This study may provide a theoretical strategy for preventing and treating the H3N8 subtype avian influenza virus-induced epidemic.METHODS:A strain of H3N8 avian influenza virus from chickens was characterised by phylogenetic analy-sis,antigenic diversity,receptor-binding specificity,neuraminidase activity,replication,and transmission in hamsters and a systematic pathological analysis was conducted.RESULTS:This novel avian influenza virus was generated through complex recombination of Eurasian avian H3 genes,North American avian N8 genes and six internal genes of H9N2 sub-type AIV.The cleavage site of the outer protein,HA,was PEKQTR↓GLF,which is characteristic of the low pathogenic avian influenza virus.The HA gene of YJ/552 exhibited the highest nucleotide homology with A/China/ZMD-22-2/2022(H3N8)at 99.09%,while the NA gene showed the highest homology with A/chicken/Dongguan/879/2022(H3N8)at 99.01%.This strain preferentially binds to avian-type receptors and could bind to human-type receptors.This virus could effectively replicate in the trachea and lungs of inoculated and contact hamsters.CONCLUSION:YJ/552 is a recombi-nant H3N8 avian influenza virus replicated in the upper respiratory system and transmitted in hamsters.This study pro-vides data support for the early warning and prevention of H3 subtype avian influenza viruses.

AIM:A strain was isolated and identified as the H3N8 subtype of the avian influenza virus from a sick chicken at a farm in Yangjiang,Guangdong Province,named A/chicken/Yangjiang/552/2023(abbreviated as YJ/552).The aim of this research is to determine its genetic evolution,biological properties and pathogenicity in hamsters.This study may provide a theoretical strategy for preventing and treating the H3N8 subtype avian influenza virus-induced epidemic.METHODS:A strain of H3N8 avian influenza virus from chickens was characterised by phylogenetic analy-sis,antigenic diversity,receptor-binding specificity,neuraminidase activity,replication,and transmission in hamsters and a systematic pathological analysis was conducted.RESULTS:This novel avian influenza virus was generated through complex recombination of Eurasian avian H3 genes,North American avian N8 genes and six internal genes of H9N2 sub-type AIV.The cleavage site of the outer protein,HA,was PEKQTR↓GLF,which is characteristic of the low pathogenic avian influenza virus.The HA gene of YJ/552 exhibited the highest nucleotide homology with A/China/ZMD-22-2/2022(H3N8)at 99.09%,while the NA gene showed the highest homology with A/chicken/Dongguan/879/2022(H3N8)at 99.01%.This strain preferentially binds to avian-type receptors and could bind to human-type receptors.This virus could effectively replicate in the trachea and lungs of inoculated and contact hamsters.CONCLUSION:YJ/552 is a recombi-nant H3N8 avian influenza virus replicated in the upper respiratory system and transmitted in hamsters.This study pro-vides data support for the early warning and prevention of H3 subtype avian influenza viruses.

Objective To investigate the gene expression profile in rat pancreatic ductal stem cells(PDSCs)when induced to differentiate into insulin-secreting cells(IPCs),with the goal of identifying key genes involved in this differentiation process.Methods The expanded PDSCs were categorized into a normal control(NC)group and an induced(Tre)group.PDSCs continued expansion culture in NC group,and cultured in induction medium for 28 days to facilitate the differentiation of PDSCs into IPCs in Tre group.Dithizone staining was employed to morphologically assess whether the cells exhibited a reddish-brown coloration,indicating a positive result.The immunofluorescence staining method was used to detect the expression of insulin(Ins)and PDX1 in the cells following induction.Additionally,ELISA was conducted to measure the Ins release from IPCs,thereby verifying the responsiveness of the induced cells to glucose-stimulated Ins secretion.Concurrently,cells were collected on induction days 0 and 28 for RNA sequencing(RNA-seq),and differentially expressed genes(DEGs)were analyzed and functionally annotated.The analysis revealed that regulatory factor X3(RFX3)was overexpressed in PDSCs,and the impact of RFX3 upregulation on differentiation induction was subsequently verified.Results Compared with NC group,DTZ staining was positive,PDX1 and Ins proteins were expressed,and an increased release of Ins in response to sugar stimulation was demonstrated in the Tre group.RNA-seq analysis identified 4270 DEGs,and functional enrichment analysis utilizing the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed associations with Ins response,positive regulation of Ins secretion,pancreatic endocrine cell development,and overall pancreatic development.Additionally,functionally related genes such as ALDHA2,CREB5,EIF6,FOXO1,RFX3,WNT5a,OGT,GPR39,SMAD6,and TRPM2 were identified,indicating involvement in the cell cycle,TGF-β1 signaling pathway,FOXO signaling pathway,and Wnt signaling pathway in the regulation of the differentiation of pancreatic ductal stem cells(PDSCs)into insulin-producing cells(IPCs).Furthermore,the upregulation of RFX3 can inhibit the expression of TGF-β1 within 72 hours,thereby promoted the formation and release of Ins from insulin-positive cells.Conclusions Multiple genes and signaling pathways associated with pancreatic β-cell function collectively regulate the differentiation of rat PDSCs into IPCs.Notably,the upregulation of RFX3 enhances this differentiation process.