ObjectiveThis study aimed to investigate the action mechanism by which Banxia Xiexintang （BXT） inhibits epithelial-mesenchymal transition （EMT） in chronic atrophic gastritis （CAG） rats by regulating the interleukin-17（IL-17）/extracellular regulated protein kinases（ERK）/CCAAT enhancer binding protein β（C/EBPβ）signaling pathway， thereby providing new theoretical evidence for the treatment of CAG with classic traditional Chinese medicine formulas. MethodsA CAG rat model was established by using the combined factor method. After successful modeling， the rats were randomly divided into the model group， low-， medium-， and high-dose groups （0.549， 1.098， 2.196 g·kg^-1， respectively） of BXT， and the positive drug group （vitacoenzyme， 0.3 g·kg^-1）. A normal control group was also set up. After 8 weeks of intervention， the pathological changes of gastric tissue were evaluated. The enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of IL-17， tumor necrosis factor-α （TNF-α）， cyclooxygenase-2 （COX-2）， and C/EBPβ in serum， as well as the contents of EMT markers in gastric mucosal tissue including E-cadherin， N-cadherin， and vimentin. The immunohistochemistry method was employed to determine the localization and protein expression levels of IL-17， p-ERK， and C/EBPβ in gastric mucosal tissue. Western blot was used to detect the protein expressions of C/EBPβ， ERK， and its phosphorylated form （p）-ERK in gastric mucosa. Real-time polymerase chain reaction （Real-time PCR） was applied to measure the mRNA expression levels of ERK， COX-2， and C/EBPβ in gastric mucosa. ResultsCompared with those in the normal control group， the rats in the model group showed gastric mucosal glandular atrophy and inflammatory cell infiltration. The protein and their related mRNA expressions of C/EBPβ， ERK， and p-ERK in gastric mucosa were significantly increased （P<0.05，P<0.01）. The levels of IL-17， TNF-α， COX-2， and C/EBPβ in serum were significantly increased （P<0.01）. The contents of N-cadherin and vimentin in gastric mucosal tissue were significantly increased， while the content of E-cadherin was significantly decreased （P<0.01）. Compared with the model group， after intervention with different doses of BXT， the pathological damage of the gastric mucosa was improved to varying degrees. The protein and mRNA expressions of C/EBPβ， ERK， and p-ERK in gastric mucosa were significantly reduced （P<0.05，P<0.01）. The levels of IL-17， TNF-α， COX-2， and C/EBP β in serum were significantly decreased （P<0.01）. The contents of N-cadherin and vimentin in gastric mucosa tissue were decreased， while the content of E-cadherin was increased （P<0.05，P<0.01）. ConclusionBXT can effectively improve the pathological damage of gastric mucosal tissue in CAG rats. Its action mechanism may be related to reducing the levels of IL-17 and TNF-α in serum， regulating the IL-17/ERK/C/EBPβ signaling pathway and inhibiting the EMT process.

Heat shock factor 1(HSF1)is a core transcription factor in cellular stress response and protein homeostasis maintenance,and is widely involved in biological processes such as cell growth,differentiation,and metabolism.This study aims to design antigenic peptides of the sheep HSF1 protein and prepare polyclonal antibodies through bioinformatics analysis and immunologi-cal techniques,and verify their application effects in sheep lung and testis tissues.Firstly,the se-quence,structure,and evolution of the sheep HSF1 gene were analyzed using bioinformatics meth-ods,and key antigenic epitopes were predicted.Then,suitable peptide fragments were selected for synthesis,emulsified with Freund's complete adjuvant or incomplete adjuvant,and used to immu-nize New Zealand rabbits.The specificity and effectiveness of the antibodies were verified by indi-rect ELISA,Western blot,and immunohistochemistry(IHC)experiments.The results showed that the HSF1 protein is 564 amino acids(aa)in length,with a molecular weight of 60.76 kDa,a theo-retical isoelectric point(pI)of 5.32,and is hydrophilic and unstable.Phylogenetic tree analysis revealed that HSF1 is highly conserved among most members of the Bovidae family,as well as in humans,mice,rats,and other species,but with local site differences.Sheep HSF1 is most closely related to goat HSF1,followed by Canadian bighorn sheep and goral.The phosphorylation sites of sheep HSF1 are predominantly concentrated in the middle and C-terminal regions,which is largely consistent with the predicted results for human HSF1,though some local differences exist.The prepared polyclonal antibodies exhibited a titer of over 1:8000 in recognizing the sheep HSF1 pro-tein,and Western blot experiments confirmed clear bands with consistent molecular weight,indica-ting high specificity of the antibodies.Furthermore,the expression and specific distribution of HSF1 were detected in sheep lung and testis tissues.This study successfully prepared polyclonal antibodies based on the sheep HSF1 antigen peptide and,for the first time,detected the immuno-histochemical distribution of HSF1 in the testis and lung tissues of Argali hybrid sheep.This pro-vides an important tool for further exploring the physiological role of HSF1 in Argali hybrid sheep and its potential applications in stress response,disease prevention,and treatment.

Objective·To investigate the differential gene expression of microglia in the gray and white matter of multiple sclerosis(MS)using single-nucleus transcriptomic analysis,aiming to explore their roles in disease progression,and identify key transcriptional regulatory networks associated with the disease.Methods·snRNA-seq data of frozen human brain tissue samples from MS patients and control individuals were obtained from the Gene Expression Omnibus(GEO)database.R language,along with R packages such as Seurat,was employed to identify cell types based on specific cell markers.Microglia were extracted from the identified cell populations and classified based on their anatomical origin,either gray matter or white matter.Dimensionality reduction and clustering techniques were utilized to identify distinct microglial subpopulations with differential characteristics.Differentially expressed genes(DEGs)between the MS and control groups at the subpopulation level were analyzed by using the Seurat package.Gene set enrichment analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)was conducted on the DEGs to further explore the biological significance of these differences.Monocle3 was used for pseudotime analysis to study dynamic changes in microglia subpopulations during disease progression.Single cell regulatory network inference and clustering(SCENIC)method was applied to analyze transcription factor(TF)regulatory networks,aiming to identify key transcription factors potentially involved in MS regulation.Results·After quality control,a total of 149 062 nuclei were retained for analysis.Following dimensional reduction and clustering,12 238 microglia were identified by using key markers,including DOCK8,CSF1R,P2RY12,and CD74.The results of GO and KEGG pathway analysis showed that in gray matter microglia,functions such as endocytosis,ion homeostasis,and lipid localization were downregulated during disease progression,while in white matter microglia,functions such as protein folding,cytoplasmic translation,and response to thermal stimuli were upregulated.SCENIC analysis revealed that the expression of transcription factors such as FLI1,MITF,and FOXP1 was upregulated in MS.Conclusion·Microglia play a critical role in MS,with white matter microglia being more significantly impacted by MS than their gray matter counterparts.Transcription factors such as FLI1,MITF,and FOXP1 are identified as key regulators involved in disease modulation,with their associated transcriptional regulatory networks playing a central role in disease modulation.

Heat shock factor 1(HSF1)is a core transcription factor in cellular stress response and protein homeostasis maintenance,and is widely involved in biological processes such as cell growth,differentiation,and metabolism.This study aims to design antigenic peptides of the sheep HSF1 protein and prepare polyclonal antibodies through bioinformatics analysis and immunologi-cal techniques,and verify their application effects in sheep lung and testis tissues.Firstly,the se-quence,structure,and evolution of the sheep HSF1 gene were analyzed using bioinformatics meth-ods,and key antigenic epitopes were predicted.Then,suitable peptide fragments were selected for synthesis,emulsified with Freund's complete adjuvant or incomplete adjuvant,and used to immu-nize New Zealand rabbits.The specificity and effectiveness of the antibodies were verified by indi-rect ELISA,Western blot,and immunohistochemistry(IHC)experiments.The results showed that the HSF1 protein is 564 amino acids(aa)in length,with a molecular weight of 60.76 kDa,a theo-retical isoelectric point(pI)of 5.32,and is hydrophilic and unstable.Phylogenetic tree analysis revealed that HSF1 is highly conserved among most members of the Bovidae family,as well as in humans,mice,rats,and other species,but with local site differences.Sheep HSF1 is most closely related to goat HSF1,followed by Canadian bighorn sheep and goral.The phosphorylation sites of sheep HSF1 are predominantly concentrated in the middle and C-terminal regions,which is largely consistent with the predicted results for human HSF1,though some local differences exist.The prepared polyclonal antibodies exhibited a titer of over 1:8000 in recognizing the sheep HSF1 pro-tein,and Western blot experiments confirmed clear bands with consistent molecular weight,indica-ting high specificity of the antibodies.Furthermore,the expression and specific distribution of HSF1 were detected in sheep lung and testis tissues.This study successfully prepared polyclonal antibodies based on the sheep HSF1 antigen peptide and,for the first time,detected the immuno-histochemical distribution of HSF1 in the testis and lung tissues of Argali hybrid sheep.This pro-vides an important tool for further exploring the physiological role of HSF1 in Argali hybrid sheep and its potential applications in stress response,disease prevention,and treatment.

BACKGROUND:Skeletal muscle insulin resistance is the key pathological link of type 2 diabetes.The traditional Chinese medicine compound Roujishuncuiyin can effectively improve skeletal muscle insulin resistance,but its mechanism has not been clarified.OBJECTIVE:To explore the mechanism of Roujishuncuiyin on skeletal muscle insulin resistance in type 2 diabetes mice.METHODS:Forty db/db mice with type 2 diabetes mellitus were randomized into a model group,a low-dose Roujishuncuiyin group,a high-dose Roujishuncuiyin group,and a positive drug group,with 10 mice in each group.The latter three administration groups were given 157.5 mg/g and 630 mg/g Roujishuncuiyin and 200 mg/g metformin hydrochloride aqueous solution by gavage once a day,respectively.In addition,10 db/dm mice were selected as the blank control group.Mice in the model and blank control groups were given the same dose of 0.9％NaCl solution by gavage.After 12 weeks of intervention,fasting blood glucose was measured in each group of mice,and oral glucose tolerance test was performed to calculate the area under the blood glucose curve.ELISA was used to detect serum insulin level and calculate the resistance index.Mitochondrial structure of skeletal muscle tissue was observed under transmission electron microscopy.Western blot was used to detect the expression levels and phosphorylation levels of protein kinase B(AKT)and glycogen synthase kinase 3β(GSK-3β)proteins in skeletal muscle.RESULTS AND CONCLUSION:(1)Compared with the blank control group,fasting blood glucose,fasting insulin and insulin resistance index were significantly higher in the model group(P＜0.05),the area under the curve of the oral glucose tolerance test was significantly increased(P＜0.05),the expression of p-AKT and p-GSK3β proteins in tibialis anterior muscle was significantly decreased(P＜0.05),and there was a large amount of mitochondrial damage in tibialis anterior muscle and a large number of lipid droplets in the interstitium.(2)Compared with the model group,fasting blood glucose,fasting insulin,and insulin resistance index were significantly reduced in the low-and high-dose Roujishuncuiyin groups and the positive control group(P＜0.05),the area under the curve of the oral glucose tolerance test was reduced(P＜0.05),the expression of p-AKT and p-GSK3β proteins in the tibialis anterior muscle was significantly elevated(P＜0.05),and mitochondrial damage in the tibialis anterior muscle was significantly ameliorated,with decreased lipid droplets in the interstitium.(3)The above indexes were better in the high-dose Roujishuncuiyin group than the low-dose Roujishuncuiyin group(P＜0.05),while there was no significant difference between the high-dose Roujishuncuiyin group and positive control group(P＞0.05).To conclude,by upregulating the protein levels of p-AKT and p-GSK3β in skeletal muscle tissue,the traditional Chinese medicine compound Roujishuncuiyin can improve structural disorders and mitochondrial morphology in skeletal muscle tissue,reduce insulin resistance in the skeletal muscle and regulate glucose homeostasis in the body.

Objective:To investigate the impact of perceived barriers to cardiac rehabilitation on adherence to cardiac rehabilitation and the mediating effect of cardiac impact denial in elderly patients after percutaneous coronary intervention (PCI) .Methods:This study was a cross-sectional survey. From August 2023 to July 2024, 336 elderly patients with PCI who were discharged from the First Affiliated Hospital of Wenzhou Medical University and needed phaseⅡ cardiac rehabilitation were selected for the study. General Information Questionnaire, Adherence of Cardiac Rehabilitation Assessment Scale in Patients with Coronary Heart Disease (ACRAS-CHD) , Cardiac Rehabilitation Barriers Scale (CRBS) , and Cardiac Denial of Impact Scale (CDIS) were used to survey the patients. Bootstrap method was used to sample 5 000 for mediating effect tests.Results:A total of 336 questionnaires were distributed and 324 valid questionnaires were recovered, with a valid recovery rate of 96.43% (324/336) . Among 324 elderly patients after PCI, ACRAS-CHD score, CRBS score, and CDIS score were (73.19±5.46) , (79.35±6.60) , and (30.57±3.66) , respectively. The direct effect of perceived barriers to cardiac rehabilitation on adherence to cardiac rehabilitation in elderly patients after PCI was -0.368, which accounted for 74.49% (-0.368/-0.494) of the total effect. The mediating effect of cardiac impact denial between perceived barriers to cardiac rehabilitation and adherence to cardiac rehabilitation in elderly patients after PCI was -0.126, which accounted for 25.51% of the total effect (-0.126/-0.494) .Conclusions:Elderly patients after PCI have poor adherence to cardiac rehabilitation. Perceived cardiac rehabilitation barriers reduce cardiac rehabilitation adherence in elderly patients after PCI, and cardiac impact denial exerts a partial mediating effect between perceived cardiac rehabilitation barriers and cardiac rehabilitation adherence.

ObjectiveTo analyze the research hotspot and future trend of non-invasive brain stimulation (NIBS) in Alzheimer's disease. MethodsRelevant literature on application of NIBS in Alzheimer's disease from January, 2014 to October, 2024 was retrieved from the Web of Science Core Collection database. CiteSpace 6.4.R1 was used to perform a bibliometric analysis and to create knowledge maps, including annual publication volume, countries, institutions, authors, keywords and co-cited references. ResultsA total of 731 articles were included, showing an increasing trend in annual publication volume. The United States was the leading country in publication volume, Harvard University was the most productive institution, and Giacomo Koch was the most prolific author. Brain Stimulation was the most frequently cited journal. Highly focused keywords included cognitive impairment, memory, dementia, transcranial magnetic stimulation and transcranial direct current stimulation. Bursting keywords in the past two years included transcranial alternating current stimulation, functional magnetic resonance imaging, brain-derived neurotrophic factor and oxidative stress. ConclusionResearch interest in NIBS within the field of Alzheimer's disease has been steadily increasing. The research hotspots include the effect and mechanism of NIBS on cognitive function and the impact of stimulating different brain regions on cognitive outcome. Future research may focus on integrating NIBS with techniques such as functional magnetic resonance imaging to achieve individualized and precise stimulation.

BACKGROUND:Skeletal muscle insulin resistance is the key pathological link of type 2 diabetes.The traditional Chinese medicine compound Roujishuncuiyin can effectively improve skeletal muscle insulin resistance,but its mechanism has not been clarified.OBJECTIVE:To explore the mechanism of Roujishuncuiyin on skeletal muscle insulin resistance in type 2 diabetes mice.METHODS:Forty db/db mice with type 2 diabetes mellitus were randomized into a model group,a low-dose Roujishuncuiyin group,a high-dose Roujishuncuiyin group,and a positive drug group,with 10 mice in each group.The latter three administration groups were given 157.5 mg/g and 630 mg/g Roujishuncuiyin and 200 mg/g metformin hydrochloride aqueous solution by gavage once a day,respectively.In addition,10 db/dm mice were selected as the blank control group.Mice in the model and blank control groups were given the same dose of 0.9％NaCl solution by gavage.After 12 weeks of intervention,fasting blood glucose was measured in each group of mice,and oral glucose tolerance test was performed to calculate the area under the blood glucose curve.ELISA was used to detect serum insulin level and calculate the resistance index.Mitochondrial structure of skeletal muscle tissue was observed under transmission electron microscopy.Western blot was used to detect the expression levels and phosphorylation levels of protein kinase B(AKT)and glycogen synthase kinase 3β(GSK-3β)proteins in skeletal muscle.RESULTS AND CONCLUSION:(1)Compared with the blank control group,fasting blood glucose,fasting insulin and insulin resistance index were significantly higher in the model group(P＜0.05),the area under the curve of the oral glucose tolerance test was significantly increased(P＜0.05),the expression of p-AKT and p-GSK3β proteins in tibialis anterior muscle was significantly decreased(P＜0.05),and there was a large amount of mitochondrial damage in tibialis anterior muscle and a large number of lipid droplets in the interstitium.(2)Compared with the model group,fasting blood glucose,fasting insulin,and insulin resistance index were significantly reduced in the low-and high-dose Roujishuncuiyin groups and the positive control group(P＜0.05),the area under the curve of the oral glucose tolerance test was reduced(P＜0.05),the expression of p-AKT and p-GSK3β proteins in the tibialis anterior muscle was significantly elevated(P＜0.05),and mitochondrial damage in the tibialis anterior muscle was significantly ameliorated,with decreased lipid droplets in the interstitium.(3)The above indexes were better in the high-dose Roujishuncuiyin group than the low-dose Roujishuncuiyin group(P＜0.05),while there was no significant difference between the high-dose Roujishuncuiyin group and positive control group(P＞0.05).To conclude,by upregulating the protein levels of p-AKT and p-GSK3β in skeletal muscle tissue,the traditional Chinese medicine compound Roujishuncuiyin can improve structural disorders and mitochondrial morphology in skeletal muscle tissue,reduce insulin resistance in the skeletal muscle and regulate glucose homeostasis in the body.

Objective:To investigate the impact of perceived barriers to cardiac rehabilitation on adherence to cardiac rehabilitation and the mediating effect of cardiac impact denial in elderly patients after percutaneous coronary intervention (PCI) .Methods:This study was a cross-sectional survey. From August 2023 to July 2024, 336 elderly patients with PCI who were discharged from the First Affiliated Hospital of Wenzhou Medical University and needed phaseⅡ cardiac rehabilitation were selected for the study. General Information Questionnaire, Adherence of Cardiac Rehabilitation Assessment Scale in Patients with Coronary Heart Disease (ACRAS-CHD) , Cardiac Rehabilitation Barriers Scale (CRBS) , and Cardiac Denial of Impact Scale (CDIS) were used to survey the patients. Bootstrap method was used to sample 5 000 for mediating effect tests.Results:A total of 336 questionnaires were distributed and 324 valid questionnaires were recovered, with a valid recovery rate of 96.43% (324/336) . Among 324 elderly patients after PCI, ACRAS-CHD score, CRBS score, and CDIS score were (73.19±5.46) , (79.35±6.60) , and (30.57±3.66) , respectively. The direct effect of perceived barriers to cardiac rehabilitation on adherence to cardiac rehabilitation in elderly patients after PCI was -0.368, which accounted for 74.49% (-0.368/-0.494) of the total effect. The mediating effect of cardiac impact denial between perceived barriers to cardiac rehabilitation and adherence to cardiac rehabilitation in elderly patients after PCI was -0.126, which accounted for 25.51% of the total effect (-0.126/-0.494) .Conclusions:Elderly patients after PCI have poor adherence to cardiac rehabilitation. Perceived cardiac rehabilitation barriers reduce cardiac rehabilitation adherence in elderly patients after PCI, and cardiac impact denial exerts a partial mediating effect between perceived cardiac rehabilitation barriers and cardiac rehabilitation adherence.

Objective·To investigate the differential gene expression of microglia in the gray and white matter of multiple sclerosis(MS)using single-nucleus transcriptomic analysis,aiming to explore their roles in disease progression,and identify key transcriptional regulatory networks associated with the disease.Methods·snRNA-seq data of frozen human brain tissue samples from MS patients and control individuals were obtained from the Gene Expression Omnibus(GEO)database.R language,along with R packages such as Seurat,was employed to identify cell types based on specific cell markers.Microglia were extracted from the identified cell populations and classified based on their anatomical origin,either gray matter or white matter.Dimensionality reduction and clustering techniques were utilized to identify distinct microglial subpopulations with differential characteristics.Differentially expressed genes(DEGs)between the MS and control groups at the subpopulation level were analyzed by using the Seurat package.Gene set enrichment analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)was conducted on the DEGs to further explore the biological significance of these differences.Monocle3 was used for pseudotime analysis to study dynamic changes in microglia subpopulations during disease progression.Single cell regulatory network inference and clustering(SCENIC)method was applied to analyze transcription factor(TF)regulatory networks,aiming to identify key transcription factors potentially involved in MS regulation.Results·After quality control,a total of 149 062 nuclei were retained for analysis.Following dimensional reduction and clustering,12 238 microglia were identified by using key markers,including DOCK8,CSF1R,P2RY12,and CD74.The results of GO and KEGG pathway analysis showed that in gray matter microglia,functions such as endocytosis,ion homeostasis,and lipid localization were downregulated during disease progression,while in white matter microglia,functions such as protein folding,cytoplasmic translation,and response to thermal stimuli were upregulated.SCENIC analysis revealed that the expression of transcription factors such as FLI1,MITF,and FOXP1 was upregulated in MS.Conclusion·Microglia play a critical role in MS,with white matter microglia being more significantly impacted by MS than their gray matter counterparts.Transcription factors such as FLI1,MITF,and FOXP1 are identified as key regulators involved in disease modulation,with their associated transcriptional regulatory networks playing a central role in disease modulation.