Abstract:BACKGROUND:Matrix metalloproteinase 9 affects osteogenic and myogenic differentiation,but its specific regulatory mechanism is not well understood. OBJECTIVE:To investigate the effects of oxidative stress injury on mitophagy in mouse MC3T3-E1 and C2C12 cells,and to examine the role of matrix metalloproteinase 9 as a critical regulator in regulating mitophagy and affecting osteogenic and myogenic differentiation via the PTEN-induced kinase 1/Parkin signaling pathway. METHODS:Mouse C2C12 and MC3T3-E1 cells were selected for differentiation and culture.Each cell line was divided into four groups:control,model(hydrogen peroxide-induced oxidative damage),GM6001(matrix metalloproteinase 9 inhibitor),and 3-methyladenine(PTEN-induced kinase 1/Parkin pathway inhibitor)groups,respectively.Cells in each group were treated for 24 hours.Flow cytometry was used to detect intracellular mitochondrial membrane potential and reactive oxygen species levels.Transmission electron microscopy was used to observe cellular mitochondrial damage and mitophagy.Western blot was used to detect matrix metalloproteinase 9 protein expression.Quantitative reverse transcription polymerase chain reaction and western blot were used to detect the mRNA and protein expressions of P62,microtubule-associated protein light chain 3,PTEN-induced kinase 1,and parkin in MC3T3-E1 cells.Western blot was used to detect the protein expressions of osteogenic proteins osteocalcin,osteopontin,and Runt-related transcription factor 2 in MC3T3-E1 cells.Quantitative reverse transcription polymerase chain reaction was used to detect the mRNA expressions of myogenic genes myogenic factor 5,myogenic factor 6,myogenic differentiation 1,and myogenin in C2C12 cells. RESULTS AND CONCLUSION:(1)In MC3T3-E1 and C2C12 cells,mitochondrial membrane potential was decreased and reactive oxygen species levels were increased in the model,GM6001,and 3-methyladenine groups compared with the control group.Reactive oxygen species levels were decreased in the GM6001 group compared with the model group,and reactive oxygen species levels were increased in the 3-methyladenine group compared with the GM6001 group.(2)Transmission electron microscopy showed that mitochondrial damage was more severe and mitophagy was formed in the model group.Mitochondrial damage was improved in the GM6001 group.Mitochondrial damage and mitophagy were weaker in the 3-methyladenine group compared with the model group.(3)Compared with the control group,matrix metalloproteinase 9 protein expression was increased in the model,GM6001,and 3-methyladenine groups.Compared with the model group,matrix metalloproteinase 9 protein expression was significantly decreased in the 3-methyladenine group.Compared with the 3-methyladenine group,matrix metalloproteinase 9 protein expression was significantly decreased in the GM6001 group.(4)Compared with the control group,the mRNA and protein expressions of P62 were decreased in the model group,further decreased in the GM6001 group,and increased in the 3-methyladenine group.Compared with the control group,the mRNA and protein expressions of microtubule-associated protein light chain 3 Ⅱ,microtubule-associated protein light chain 3 Ⅰ,PTEN-induced kinase 1,and Parkin were elevated in the model group,further elevated in the GM6001 group,and decreased in the 3-methyladenine group.(5)In the MC3T3-E1 cells,compared with the control group,the protein expressions of osteocalcin,osteopontin,and Runt-related transcription factor 2 were decreased in the model,GM6001,and 3-methyladenine groups.Compared with the model group,the protein expressions of osteocalcin,and osteopontin were increased in the GM6001 group.Compared with the GM6001 group,the protein expressions of osteocalcin and osteopontin were decreased in the 3-methyladenine group.(6)In the C2C12 cell line,compared with the control group,the mRNA expressions of myogenic factor 5,myogenic factor 6,myogenic differentiation 1,and myogenin were decreased in the model,GM6001,and 3-methyladenine groups.Compared with the model group,the expressions of the above myogenic genes were elevated in the GM6001 group,and compared with the GM6001 group,the above myogenic gene expressions were decreased in the 3-methyladenine group.The results showed that matrix metalloproteinase 9 exhibits high expression levels in oxidative injury models,and its inhibitors enhance mitophagy in the oxidative damage environment.This regulation was based on the activation of PTEN-induced kinase 1/Parkin pathway,thereby promoting osteogenic and myogenic differentiation.

Abstract:BACKGROUND:The Chinese medicine Compound Dandeng Tongnao Soft Capsule is characterized by the synergistic effect of multiple components,but traditional research methods are difficult to systematically reveal its complex mechanism of action.This experiment innovatively integrates high-performance liquid chromatography fingerprinting and network pharmacology methods.This integrated strategy can not only achieve a comprehensive evaluation of the material basis of traditional Chinese medicine,but also further elucidate the targets of traditional Chinese medicine,clarify its pathways and mechanisms,and provide a new research paradigm for elucidating the"multi-component-multi-target-multi-pathway"mechanism of traditional Chinese medicine prescriptions. OBJECTIVE:To investigate the material basis and mechanism of action of Dandeng Tongnao Soft Capsules against ischemic stroke based on HPLC fingerprinting and network pharmacology. METHODS:The fingerprints of 10 batches of Dandeng Tongnao Soft Capsules were determined by high-performance liquid chromatography,and the common peaks in the fingerprints of Dandeng Tongnao Soft Capsules were identified by LCMS-IT-TOF technique.Common peak component targets of Dandeng Tongnao Soft Capsules were predicted by SwissTargetPrediction database(developed by Swiss Institute of Bioinformatics;function:prediction of potential targets based on the structure of compounds).Ischemic stroke-related targets were collected through the OMIM database(maintained by Johns Hopkins University,USA;function:to include human genetic diseases and disease-causing genes)and the GeneCards database(developed by the Weizmann Institute of Science,Israel;function:to integrate information on gene functions,disease associations and pathways).Drug-disease intersection targets were screened using the Venny 2.1.0 online tool to draw Venn diagrams.Cytoscape 3.10.1 software was used to construct a"drug-compound-target"interaction network to screen the potential components of Dandeng Tongnao Soft Capsules against ischemic stroke.Protein-protein interaction network analysis was performed through the STRING database(jointly developed by the European Molecular Biology Laboratory and other institutions,function:analysis of protein interaction networks and core targets)to obtain core targets.DAVID database(developed by National Institute of Allergy and Infectious Diseases,Function:Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis)was used to analyze the biological functions and pathways of the core targets of Dandeng Tongnao Soft Capsules against ischemic stroke,and ultimately to build a multi-dimensional network of"drug-component-target-pathway-disease." RESULTS AND CONCLUSION:(1)The fingerprint pattern of Dandeng Tongnao Soft Capsules was established,24 common peaks were calibrated,the peaks were well separated,and the similarity between samples was greater than 0.9.Twenty-four peaks in Dandeng Tongnao Soft Capsule were identified.A total of 508 intersecting targets of components and diseases in Dandeng Tongnao Soft Capsule were obtained.(2)Protein-protein interaction analysis,GO function enrichment and KEGG pathway enrichment analysis revealed that Dandeng Tongnao Soft Capsules may exert anti-ischemic stroke effects through its active ingredients,geranodendronin,danshenone I,tanshenone IIA,ethyl ferulate,and tanshenaldehyde,which act on glyceraldehyde-3-phosphate dehydrogenase,serine/threonine protein kinase 1,and tumor necrosis factor,non-receptor tyrosine kinase,to regulate hypoxia-inducible factor-1 signaling pathway,advanced glycosylation end product-receptor signaling pathway,and phosphatidylinositol 3-kinase protein kinase B signaling pathway.To conclude,the organic integration of chemical analysis and bioinformatics prediction systematically explains the multidimensional mechanism of action of Dandeng Tongnao Soft Capsules.It not only provides a scientific basis for its clinical efficacy,but also,more importantly,establishes an integrated research framework of"fingerprinting-network pharmacology,"which provides a general methodological reference for the comprehensive evaluation of the material basis of traditional Chinese medicines and the analysis of their mechanism of action,and has important theoretical and practical value for the modernization of traditional Chinese medicines.

Abstract:BACKGROUND:Youth is a critical window for physical development in the human body.During this stage,optimization of skeletal muscle function—including muscle strength,power output,and muscular endurance—is not only essential for athletic performance but also serves as a frontline defense against muscle decline and metabolic disorders caused by modern lifestyles.Meanwhile,cardiorespiratory endurance reflects the overall function of the cardiovascular and respiratory systems,which is a key predictor of health risks. OBJECTIVE:To investigate the effects of different arrangement order of strength and endurance training(strength priority vs.endurance priority)on lower limb skeletal muscle function(maximum strength and strength endurance)and aerobic metabolic capacity(maximum oxygen uptake)in young people. METHODS:Thirty healthy young college students from Hebei Sport University were randomly divided into three groups:strength priority group(n=10),endurance priority group(n=10)and control group(n=10).The strength priority group received strength training followed by endurance training,the endurance priority group had the opposite order,and the control group maintained daily activities.The intervention in each group lasted for 8 weeks,twice a week.The one repetition maximum(1RM)in squat/deadlift of the lower limb(maximum strength),the maximum repetitions at 70%1RM squat/deadlift(strength endurance),and maximal oxygen uptake(estimated by indirect methods)were measured before and after the intervention. RESULTS AND CONCLUSION:After 8 weeks of intervention,1RM squat/deadlift were significantly increased in strength priority group(P<0.05,P<0.01),and the maximum repetitions at 70%1RM squat/deadlift were significantly increased(P<0.01).In the endurance priority group,the maximum repetitions at 70%1RM squat/deadlift were significantly increased(P<0.01),but there was no significant change in 1RM squat/deadlift.The maximum oxygen uptake increased significantly in the endurance priority group(P<0.05),remained unchanged in the strength priority group,and decreased significantly in the control group(P<0.05).There were significant differences in the maximum repetitions at 70%1RM squat/deadlift and maximum oxygen uptake between groups(P<0.01),and the maximum oxygen uptake in the endurance priority group was significantly higher than that in the strength priority group(P<0.05)and control group(P<0.01).The above results suggest that,during synchronic training,the priority of strength training is more conducive to improving the maximum strength of the lower limbs and maintaining aerobic capacity in young people,while the priority of endurance training is more conducive to improving the strength endurance and aerobic metabolic capacity.

Abstract:BACKGROUND:Deep brain stimulation of the anterior cingulate cortex has emerged as a new surgical treatment for psychiatric disorders such as depression;however,the specific mechanisms underlying its therapeutic effects are still not well understood.It has been shown that damage to the anterior cingulate cortex improves depressive behavior in mice,leading to the hypothesis that deep brain stimulation may hold promise as a treatment for depression. OBJECTIVE:To elucidate the neuroregulatory mechanisms of DBS of the anterior cingulate cortex and compares the effects of DBS and electroablation on depression-like behavior in mice using calcium imaging and fiber optic recording combined with C-fos immunohistochemistry. METHODS:(1)Grouping and modeling:C57BL/6J mice were randomly divided into four groups:a control group,a model+sham stimulation group,a model+deep brain electrical stimulation group,and a model+electroablation group.Chronic restraint stress(3.5 weeks)was conducted to establish mouse models of depression in the latter three groups,and deep brain electrical stimulation electrodes were implanted in the anterior cingulate cortex in all four groups.(2)Intervention protocol:Mice in the deep brain stimulation group received daily 2-hour high-frequency electrical stimulation(130 Hz,200 μA,50 μs)for 1 week;mice in the model+sham stimulation group received electrode implantation without electrical stimulation;mice in the model+ablation group received a single 0.5-hour alternating current stimulation(130 Hz,200 μA).(3)Behavioral assessment:Depression-like behavior was quantified using the forced swim test and tail suspension test.(4)Circuit mechanism analysis:Real-time neural activity was monitored by fiber photometry to assess the activation effect of deep brain stimulation on the anterior cingulate cortex-basolateral amygdala circuit.(5)Anxiety behavior analysis:Open field test was performed to assess anxiety-like behavior in mice after intervention.(6)Whole-brain activation map:C-fos immunohistochemical staining was used to quantify neuronal activity in the downstream brain regions of the anterior cingulate cortex.(7)The improvement in depression-like behavior in mice was verified using electroablation. RESULTS AND CONCLUSION:(1)Behavioral analysis:The forced swim test showed that the immobility time was significantly longer in the model+deep brain stimulation and model+sham stimulation groups than the control group(P<0.05),but there was no significant difference between the model+deep brain stimulation and model+sham stimulation groups;the immobility time was significantly shorter in the model+electroablation group than that of the model+sham stimulation group(P<0.01).In the tail suspension test,there was no significant difference in immobility time between the model+deep brain stimulation and model+sham stimulation groups.In the open field test,the central zone residence time and locomotor distance were significantly shorter in the deep brain stimulation group than the control group(P<0.05),suggesting that deep brain stimulation may exacerbate anxiety-like behavior.(2)Neural mechanisms:Fiber optic recording confirmed that anterior cingulate cortex-deep brain stimulation specifically activated the anterior cingulate cortex-basolateral amygdala circuit.(3)C-fos staining showed that deep brain stimulation significantly enhanced neuronal activity in the downstream brain regions of the anterior cingulate cortex.These results suggest that although traditional high-frequency anterior cingulate cortex-deep brain stimulation effectively activates the target neural circuit,it fails to improve depression-like behavior and may exacerbate anxiety states by enhancing limbic system activity;conversely,functional inhibition of the anterior cingulate cortex by electroablation exhibits marked antidepressant effects,providing new directions for optimizing neuromodulation strategies.

Abstract:BACKGROUND:Spinal cord injury leads to varying degrees of muscle atrophy.While there are multiple treatment options available for muscle atrophy after spinal cord injury,few studies have compared the efficacy of different rehabilitative strategies in addressing muscle atrophy resulting from spinal cord injury. OBJECTIVE:To investigate the effects of neuromuscular electrical stimulation,repetitive peripheral magnetic stimulation and treadmill training on gastrocnemius muscle function in rats after spinal cord injury. METHODS:Thirty female Sprague-Dawley rats were randomly divided into five groups,including the sham-operated group,the model group,the electrical stimulation group,the magnetic stimulation group,and the treadmill training group.The rats in the sham-operated group only underwent spinous process and laminar dissection without spinal cord injury,while rats in the other groups were subjected to a modified Allen's method to establish a T10 incomplete moderate spinal cord injury model.After modeling,the sham-operated group and the model group received no intervention,whereas the remaining groups underwent corresponding bilateral gastrocnemius neuromuscular electrical stimulation,repetitive peripheral magnetic stimulation,or treadmill exercise training.The Basso,Beattie and Bresnahan locomotor scale and inclined plate test were used to assess the hind limb motor function of the rats before and 0,3,7,and 14 days after modeling.The gastrocnemius muscle tissues were collected on the 14th day after surgery,and the muscle atrophy was assessed by relative wet weight ratio of the gastrocnemius muscle.The cross-sectional area of muscle fibers of the gastrocnemius muscle was detected by hematoxylin-eosin staining.The distribution and activity of acetylcholinesterase at the neuromuscular junction of the gastrocnemius muscle were detected by acetylcholinesterase staining.The expression of Atrogin-1 and MuRF1 protein in gastrocnemius muscle was detected by western blot assay. RESULTS AND CONCLUSION:(1)Compared with the sham-operated group,the Basso,Beattie and Bresnahan score,inclined angle of the inclined plate test,relative wet weight ratio of the gastrocnemius muscle,muscle fiber cross-sectional area,and average absorbance value of acetylcholinesterase in the gastrocnemius muscle were significantly decreased in the model group(P<0.05),and the expression of Atrogin-1 and MuRF1 protein was significantly increased(P<0.05).(2)Compared with the model group,the Basso,Beattie and Bresnahan score,inclined angle of the inclined plate test,relative wet weight ratio of the gastrocnemius muscle,muscle fiber cross-sectional area,and average absorbance value of acetylcholinesterase in the gastrocnemius muscle were significantly increased in the electrical stimulation,magnetic stimulation,and treadmill training groups(P<0.05).Moreover,the above indicators in the treadmill training group were significantly higher than those in the magnetic stimulation group,while the Basso,Beattie and Bresnahan score,inclined angle of the inclined plate test,relative wet weight ratio of the gastrocnemius muscle,and average absorbance value of acetylcholinesterase in the gastrocnemius muscle in the treadmill training group were significantly higher those in the electrical stimulation group(P<0.05).(3)In addition,the expression level of Atrogin-1 protein was significantly lower in the electrical stimulation,magnetic stimulation,and treadmill training groups than in the model group,while the expression level of Atrogin-1 protein in the treadmill training group was significantly lower than that in the magnetic stimulation group(P<0.05).The expression level of MuRF1 protein was significantly lower in the electrical stimulation group and the treadmill training group than in the model group(P<0.05),while there was no significant difference in the protein expression of MuRF1 between the magnetic stimulation group and the model group(P>0.05).To conclude,neuromuscular electrical stimulation,repetitive peripheral magnetic stimulation and treadmill training can all improve gastrocnemius muscle function and reduce muscle atrophy in rats with spinal cord injury,with treadmill training showing the best effects.

Abstract:BACKGROUND:Recent advancements in brain-computer interface technology have demonstrated its clinical efficacy in stroke rehabilitation,yielding substantial therapeutic outcomes.A comprehensive visual analysis is imperative to elucidate current research frontiers and identify emerging hotspots in this rapidly evolving field. OBJECTIVE:To systematically examine research frontiers and developmental trends in brain-computer interface applications for stroke rehabilitation using bibliometric visualization tools. METHODS:Based on the Web of Science Core Collection and China National Knowledge Infrastructure(CNKI)databases,Citespace 6.4.1,VOSviewer 1.6.20,and Excel 2021 were employed for visualized data analysis of retrieved Chinese and English literature focusing on the application of brain-computer interface technology in post-stroke functional recovery.Using scientometric methods,we conducted an in-depth analysis of the current research status,hot topics,and future trends regarding brain-computer interface technology in stroke rehabilitation. RESULTS AND CONCLUSION:(1)A total of 985 Chinese and English articles(879 in English and 106 in Chinese)published between 2003 and 2025 were included.Annual publication output in this field has shown a consistent growth trend both domestically and globally.(2)China,the United States,and Germany were the most productive countries.The University of Tübingen(Germany)ranked as the most influential institution,while Huashan Hospital,Fudan University,topped Chinese institutions in publication output.Frontiers in Neuroscience(Switzerland)was the leading English journal,and the Chinese Journal of Rehabilitation Medicine was the foremost Chinese journal.Birbaumer Niels(Germany)had the highest publication output in English,whereas Jia Jie was the most prolific author in Chinese literature.(3)International research focused on theoretical validation and clinical efficacy,with a particular emphasis on upper limb and neural recovery.Domestic studies prioritized technological and systemic optimization,with a focus on exploring broader applications in rehabilitation.(4)"Motor imagery"emerged as a common high-frequency keyword in both Chinese and English literature,with research hotspots focusing on electroencephalography-based and motor imagery-driven brain-computer interface systems.(5)Future trends are likely to include multimodal integration,artificial intelligence fusion,expanded rehabilitation approaches,and strengthened international collaboration.

Abstract:BACKGROUND:Cuproptosis,as an emerging form of cell death,has garnered widespread attention in recent years.However,there is a lack of systematic understanding of the global research dynamics of cuproptosis,especially the research priorities and trends in different countries and academic communities.A multi-database,multi-faceted bibliometric analysis is urgently needed to provide a more comprehensive understanding of the research progress in the field of cuproptosis. OBJECTIVE:To systematically analyze the research status,hotspots,and development trends in the field of cuproptosis using bibliometric methods. METHODS:Using bibliometric methods combined with CiteSpace and VOSviewer visualization and analysis tools,we retrieved and analyzed 1 378 cuproptosis-related literature published from 2022 to 2024 in the international authoritative Web of Science core collection database and the China Knowledge Network database(CNKI).This analysis included annual publication volume,research cooperation network,keyword co-occurrence,clustering and hotspot change characteristics,and highly cited literature.By comparing and analyzing the data from these two databases,we aim to reveal the overall landscape of cuproptosis research from a global perspective. RESULTS AND CONCLUSION:Cuproptosis research has shown a rapid growth trend,with China leading in the number of publications,where institutions such as Shanghai Jiao Tong University and Central South University serve as the core institutions.The research theme focuses on the mechanism of cuproptosis,tumor therapeutic application(chemokinetic therapy)and cross-regulation with ferroptosis.The keyword emergence analysis showed that"immune microenvironment"and"tumor therapy"are the research hotspots.Machine-learning gene prediction models and single-cell sequencing technologies are driving the development of precision medicine.Most of the English studies focus on the exploration of mechanisms and application of technologies.Their research methods,technical routes and hot directions are of great significance to the cuproptosis research in China,which is in the rapid development period.In particular,interdisciplinary cooperation and multi-center clinical translational research in the field of tumor treatment and other major diseases are expected to provide more targeted and effective new strategies for the diagnosis and treatment of malignant tumors and other major diseases,including the Chinese population.

Abstract:BACKGROUND:Epstein-Barr virus(EBV),a human herpesvirus,exhibits widespread prevalence in the global population.Previous observational studies have suggested an association between EBV infection and ankylosing spondylitis;however,conventional methodologies are limited by confounding factors and reverse causation bias,precluding definitive causal inference.Elucidating the causal relationship between EBV and ankylosing spondylitis is critical for uncovering the immune pathogenesis of ankylosing spondylitis and establishing a theoretical foundation for EBV-targeted preventive strategies. OBJECTIVE:To investigate the bidirectional causal relationship between EBV infection and ankylosing spondylitis using bidirectional Mendelian randomization(MR)analysis. METHODS:Bidirectional two-sample MR analysis was conducted using European population-wide genome-wide association study(GWAS)summary statistics.The generalized summary data MR approach was integrated with conventional methods,including inverse-variance weighted,MR-Egger,and weighted median estimators,to assess bidirectional causality between EBV infection and ankylosing spondylitis.EBV antibody data(EA-D,EBNA-1,VCA p18,and ZEBRA)were sourced from the UK Biobank,while ankylosing spondylitis data were obtained from FinnGen.Instrumental variables were selected based on genome-wide significance(P<5×10⁻⁶),with exclusion of linkage disequilibrium and confounding single-nucleotide polymorphisms associated with smoking,rheumatoid arthritis,or psoriasis.Statistical significance was determined using Bonferroni correction(threshold:P=6.3×10-3),supplemented by sensitivity analyses for heterogeneity(Cochran's Q),pleiotropy(MR-Egger intercept,MR-PRESSO),and robustness(leave-one-out analysis).The generalized summary data MR approach further applied HEIDI-outlier testing(P<0.01)to eliminate pleiotropic single-nucleotide polymorphisms,ensuring causal inference validity. RESULTS AND CONCLUSION:(1)Bidirectional MR revealed that EBV infection significantly increased the risk of ankylosing spondylitis:Elevated EBNA-1 antibody levels showed a positive association with ankylosing spondylitis[odds ratio(OR)=1.41,95%confidence interval(CI):1.14-1.76,P=0.002],while ZEBRA antibodies demonstrated a stronger effect(OR=1.56,95%CI:1.31-1.85,P=5.4×10⁻⁷),suggesting both latent(EBNA-1)and lytic(ZEBRA)EBV phases may drive ankylosing spondylitis pathogenesis via cross-reactive immune responses.(2)Reverse MR analysis indicated an inverse correlation between ankylosing spondylitis and EBV lytic-phase EA-D antibodies(OR=0.96,95%CI:0.94-0.98,P=3.25×10⁻⁴),indicating that ankylosing spondylitis-associated immune profiles may suppress EBV reactivation.(3)All results remained robust in heterogeneity,pleiotropy,and sensitivity analyses,with no detectable bias.(4)This study is the first to confirm EBV infection as an independent causal risk factor for ankylosing spondylitis based on international databases and European population data.Although genetic heterogeneity exists across populations,these findings in European populations provide critical insights into shared immune mechanisms underlying ankylosing spondylitis.Future validation in Chinese cohorts is warranted to characterize EBV-ankylosing spondylitis molecular interactions specific to this population.Furthermore,the generalized summary data MR framework exemplifies the utility of public GWAS data for causal inference,offering a paradigm for efficient risk factor discovery and helping the development of precision medicine.

Abstract:BACKGROUND:Currently,behavioral tests for assessing learning and memory are important technical methods for the drug development and exploration of the pathogenesis of Alzheimer's disease.However,animal behavior is easily influenced by various factors,including external conditions,the function of internal organs and cells,and psychological states.Therefore,developing appropriate experimental methods that can effectively reflect animal cognitive behavior is an important prerequisite for exploring the pathogenic mechanism and prevention and treatment of Alzheimer's disease. OBJECTIVE:To verify that the treadmill test is a new method using two different mouse models and to explore the advantages and disadvantages of the treadmill test in evaluating the learning and memory ability of mice in comparison with the Morris water maze. METHODS:The genotypes of APP/PS1 transgenic mice(Alzheimer's disease group,n=8)and littermate wild-type mice(control group,n=8)and the pathological features of β-amyloid in the brain were identified using polymerase chain reaction and thioflavin S staining.Subsequently,11-month-old APP/PS1 transgenic mice(Alzheimer's disease group,n=8)and their littermates of wild-type mice(control group,n=8)were subjected to a treadmill test.At the same time,8-month-old 3×Tg-AD mice(Alzheimer's disease group,n=8)and wild-type control mice(control group,n=8)were subjected to treadmill and water maze tests sequentially. RESULTS AND CONCLUSION:(1)The polymerase chain reaction results showed that the expression of APP and PSEN1 genes in APP/PS1 transgenic mice was higher than that in the control group(P<0.001),and thioflavin S staining showed that there were more β-amyloid plaques in the hippocampus of APP/PS1 transgenic mice than in the control group(P<0.001).The results of the treadmill test showed that the total number and duration of electric shocks in APP/PS1 transgenic mice were higher than those in the control group(P<0.05).(2)The results of the treadmill test showed that the total number and duration of electric shocks in 3×Tg AD mice were higher than those in the control group(P<0.05).The results of the water maze test showed that the escape latency of 3×Tg AD mice was longer than that of the control group from the 4th day of the localization navigation phase(P<0.05).To conclude,the treadmill test can effectively reflect the learning and memory function of mice,and may become an experimental method for measuring mouse cognitive behaviors.

Abstract:BACKGROUND:Protein S-palmitoylation is a reversible post-translational lipid modification that directly influences protein stability,subcellular localization and interactions with other molecules.It is catalyzed in mammalian cells by 23-24 palmitoyl acyltransferases(PATs)that share the common Asp-His-His-Cys(DHHC)motif.Recent studies have highlighted the important role of S-palmitoylation in disease,and increasing studies have attempted to explore new therapeutic potentials in diverse diseases,driving efforts to target palmitoylation and its catalytic enzymes.Among these,palmitoyl transferase 5(ZDHHC5)is a more specific member of the palmitoyl group transferase family,which is a plasma membrane-localized PAT that palmitoylates a wide variety of substrates,thereby influencing multiple biological processes. OBJECTIVE:To provide an overview of the biochemical process of palmitoylation and its detection methods,to elucidate the role of ZDHHC5 in physiological processes such as cytoplasmic division,synapse formation and plasticity,and programmed cell death,and to summarize recent research progress on ZDHHC5 in cancer,respiratory diseases and other pathologies. METHODS:The first author conducted a systematic literature search in PubMed and CNKI databases for relevant literature on ZDHHC5 research in recent years,using"ZDHHC5,DHHC5 or palmitoyl transferase 5"as the search terms in English and"palmitoyl transferase 5"as the search term in Chinese.The search period was set from January 2011 to March 2025.After screening the retrieved literature for relevance and quality,the remaining literature was used to analyze the role and mechanism of ZDHHC5 in tissue homeostasis and diseases. RESULTS AND CONCLUSION:ZDHHC5,a key palmitoyl transferase that dynamically modifies protein palmitoylation,regulates cell division and differentiation,ion flux regulation,synapse formation and plasticity,autophagy and programmed cell death,intracellular membrane transport,cell adhesion,oligodendrocyte and myelin sheath formation,and immune signaling regulation,either by direct modification of key substrates,such as PCDH7,TrpM7,δ-catenin,NCX1,NOD2,and MLKL,or by formation of a complex with proteins,such as GOLGA7.Under pathological conditions,the aberrant expression of ZDHHC5 may promote the development of glioma,pancreatic cancer,lung cancer and other cancers by affecting the palmitoylation levels of EZH2,SSTR5,and INCENP and other proteins.In addition,ZDHHC5 promotes the occurrence and development of various diseases such as respiratory disease,schizophrenia,heart disease,inflammation,fatty liver,diabetic retinopathy,suggesting that its aberrant function may play an important role in the pathogenesis of multiple diseases.Future studies should focus on the substrate-specific recognition mechanism of ZDHHC5 and the regulation of its function under different physiological and pathological conditions.Meanwhile,the development of small molecule inhibitors targeting ZDHHC5 and the integrated application of interdisciplinary technologies will provide a new direction for precision therapeutic strategies based on palmitoylated modifications,which will ultimately promote the diagnosis and treatment of related diseases.

Abstract:BACKGROUND:Rheumatoid arthritis is a complex autoimmune disease,which involves the interaction of many factors including heredity,environment and immunity.Traditional research focuses on the relationship between individual protein and rheumatoid arthritis,but lacks systematic evaluation of the causal relationship between large-scale plasma proteins and rheumatoid arthritis.Mendelian randomization,as an innovative causal inference method,uses genetic variation as a tool variable,which avoids the interference of confounding factors in traditional epidemiological research and provides a new way to reveal the causal relationship between plasma protein and rheumatoid arthritis. OBJECTIVE:To study the potential relationship between human plasma protein and rheumatoid arthritis,and to explore the possible biological pathways involved in the pathogenesis of rheumatoid arthritis. METHODS:The public data of plasma protein(exposure factor)and rheumatoid arthritis(outcome factor)were collected.Among them,the data of 4 907 types of plasma proteins came from genome-wide association studies published in the journal Nature Genetics in 2022,which is now included in the Icelandic deCODE Genetics database.The study covered 35 559 Icelanders and found 18 084 protein quantitative trait loci.Genome-wide association study data of rheumatoid arthritis came from FinnGen database(R11 version),including 14 818 cases of rheumatoid arthritis and 287 796 cases in the control group.All participants were from the European population.The potential causal relationship between plasma protein and rheumatoid arthritis was evaluated by two-way Mendelian randomized analysis.Mendelian randomized analysis included five analysis methods:inverse variance weighted method,MR-Egger regression method,weighted median method,weighted model method and simple model method.Heterogeneity test,horizontal pleiotropy test and leave-one-out analysis were used to test whether the results of Mendelian randomized analysis are reliable.Reverse Mendelian randomization analysis took the plasma protein that met the screening conditions in the positive Mendelian randomization analysis as the outcome for effect analysis and sensitivity analysis. RESULTS AND CONCLUSION:(1)The results of inverse variance weighting method showed that plasma proteins CRAT and LYG1 were directly and positively correlated with rheumatoid arthritis.Plasma proteins FAM177A1,JPH4 and SPAG11B had a direct negative causal relationship with rheumatoid arthritis.(2)Reverse Mendelian randomization results showed that rheumatoid arthritis had a direct positive causal relationship with LYG1.(3)No heterogeneity or horizontal pleiotropy was found in two-way sensitivity analysis.(4)With plasma proteins as the exposure and rheumatoid arthritis as the outcome,two plasma proteins CRAT and LYG1 were positively correlated with rheumatoid arthritis,and three plasma proteins FAM177A1,JPH4 and SPAG11B were negatively correlated with rheumatoid arthritis.With rheumatoid arthritis as the exposure and plasma proteins CRAT,LYG1,FAM177A1,JPH4,SPAG11B as the outcome,rheumatoid arthritis had a positive causal relationship with the plasma protein LYG1.The above results suggest that there may be causal relationship between these five plasma proteins and rheumatoid arthritis,which will help the development of potential therapeutic and monitoring targets for rheumatoid arthritis in the future.Although the protein targets found in European databases need to be verified in the China population,the preliminary evidence of such causal associations can guide domestic research to give priority to related protein pathways and accelerate targeted drug research and development.At the same time,it calls for strengthening the construction of a local multi-center biological sample bank and a proteomics-genome-wide association studies integrated platform to overcome the limitations of European databases and promote precision medical research based on Asian populations.

Abstract:BACKGROUND:Rotator cuff tears are a prevalent condition affecting the shoulder joint,which can substantially diminish a patient's quality of life.Surface electromyography is a diagnostic technique that captures the electrophysiological signals of muscles during movement.Most of its studies have focused on special populations such as athletes and hemiplegic patients,but its use in patients with rotator cuff tears is still rare. OBJECTIVE:To compare the differences in the movement patterns of the muscles around the scapula between normal subjects and patients with rotator cuff tears using surface electromyography and to investigate the pathogenetic mechanisms of scapular dyskinesia in patients with rotator cuff tears,thereby providing a basis for the establishment of clinical rehabilitation protocol. METHODS:From September 2021 to February 2023,40 patients with rotator cuff tears who met the enrolled criteria at Guanghua Hospital affiliated to Shanghai University of Chinese Medicine were included in the trial group,and a control group without shoulder disease(n=40)was set.Demographic and clinical data were obtained using surface electromyography.Using the maximal voluntary isometric contraction(MVIC)method,the%MVIC of the upper trapezius,middle trapezius,lower trapezius,and serratus anterior muscles was measured during shoulder flexion,abduction,and external rotation at the side.Each movement was tested thrice,and the average%MVIC ratios of upper trapezius/lower trapezius,upper trapezius/middle trapezius,and upper trapezius/serratus anterior were calculated for intergroup comparison. RESULTS AND CONCLUSION:Compared with the control group,the trial group showed significantly higher%MVIC ratios of upper trapezius/lower trapezius,upper trapezius/middle trapezius,and upper trapezius/serratus anterior during shoulder flexion,abduction,and external rotation at the side(all P<0.05).To conclude,the mechanism of scapular dyskinesis in patients with rotator cuff tears may involve overactivation of the trapezius and serratus anterior muscles,resulting in movement imbalance.

Abstract:BACKGROUND:Studies have demonstrated that specific microRNAs(miRNAs)exhibit significant downregulation in a rat model of premature ovarian failure,and their overexpression can effectively alleviate symptoms of premature ovarian failure.Therefore,investigating miRNAs as potential auxiliary biomarkers holds substantial academic and clinical significance,offering a more stable and reliable reference for the diagnosis of premature ovarian failure. OBJECTIVE:To investigate whether the expression patterns of six miRNAs in ovarian tissue from animal models of premature ovarian failure and in peripheral blood from patients with premature ovarian failure are consistent,and to validate the clinical diagnostic potential of these six miRNAs. METHODS:(1)Twenty Wistar rats were randomly divided into a normal group(n=10)and a premature ovarian failure group(n=10).The model of premature ovarian failure was established in the premature ovarian failure group by intraperitoneal injection of cisplatin[1 mg/(kg·d)]for 14 continuous days.Following modeling,serum samples were collected from both groups.Levels of follicle-stimulating hormone,estradiol,and anti-Müllerian hormone were measured using ELISA.Ovarian tissue morphology was examined via hematoxylin-eosin staining.Q-PCR was used to detect the expression of miR-10a-5p,miR-21-5p,miR-22-3p,miR-126-3p,miR-144-3p,and miR-144-5p in ovarian tissue.(2)Ten patients with premature ovarian failure and ten healthy female controls,all aged<40 years,were enrolled.Chemiluminescence assays measured the levels of follicle-stimulating hormone,estradiol,and anti-Müllerian hormone in peripheral blood from both groups.Q-PCR was used to detect the expression of miR-10a-5p,miR-21-5p,miR-22-3p,miR-126-3p,miR-144-3p,and miR-144-5p in peripheral blood.The effect size and diagnostic performance of these six miRNAs in human blood samples were systematically evaluated using receiver operating characteristic curves. RESULTS AND CONCLUSION:(1)Compared with the normal group,rats with premature ovarian failure showed significantly elevated follicle-stimulating hormone level and reduced estradiol and anti-Müllerian hormone levels.Hematoxylin-eosin staining revealed a significant decrease in the number of follicles in the ovaries of rats with premature ovarian failure compared with the normal group.The expression of miR-10a-5p,miR-21-5p,miR-22-3p,miR-126-3p,miR-144-3p,and miR-144-5p was markedly downregulated in the ovaries of rats with premature ovarian failure compared with the normal group(P<0.05).(2)Compared with healthy controls,patients with premature ovarian failure exhibited elevated levels of follicle-stimulating hormone in peripheral blood(P<0.05),along with decreased levels of estradiol and anti-Müllerian hormone(P<0.05).Q-PCR assay revealed that,compared with healthy controls,patients with premature ovarian failure exhibited downregulated expression of miR-10a-5p,miR-21-5p,miR-22-3p,miR-126-3p,miR-144-3p,and miR-144-5p in peripheral blood(P<0.05).The receiver operating characteristic curves demonstrated that all six miRNAs exhibited excellent diagnostic discrimination capabilities(sensitivity:100%,specificity:100%),indicating their clinical potential as highly specific molecular biomarkers for premature ovarian failure.These findings indicate that these six miRNAs may serve not only as promising biomarkers for premature ovarian failure but also as clinically valuable tools.

Abstract:BACKGROUND:Inflammaging,characterized by a chronic low-grade pro-inflammatory state during natural aging,is closely associated with the development of age-related diseases such as cardiovascular disorders.However,a systematic theoretical framework centered on cardiac inflammaging as a core pathological mechanism is still lacking. OBJECTIVE:To analyze the prescription patterns and medication principles for treating cardiac aging in traditional Chinese medicine(TCM)classics using data mining and network pharmacology methods,and to explore the potential mechanisms of core drugs. METHODS:(1)Formulations for treating cardiac aging were collected and compiled from the 2020 edition of the Pharmacopoeia of the People's Republic of China,the 10th edition of Chinese Materia Medica,and the Comprehensive Dictionary of TCM Prescriptions.The frequency of medicinal herbs,their properties,and meridian affinity were statistically analyzed,along with association rule analysis and clustering,to identify core herb pairs for treating cardiac aging.A core herb pair-active ingredient-disease-common target network model was established,followed by gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis.In addition,molecular docking and dynamic simulation were employed to assess the binding ability and stability between potential key targets and active ingredients.(2)Validation experiment:Sixty C57BL/6 mice were randomly divided into three intervention groups:the blank group(n=20)was administered distilled water via gavage,the low-dose musk-aconite group(n=20)was administered 2.64 g/kg musk-aconite via gavage,and the high-dose musk-aconite group(n=20)was administered 5.28 g/kg musk-aconite group via gavage,once daily for 7 consecutive days.After the final administration,blood was collected from the orbital sinus,and serum was separated to obtain drug-free serum and low-and high-dose drug-containing serum of musk-aconite.Passage 3 human ventricular cardiomyocyte line AC16 was divided into three groups and treated with drug-free serum and low-and high-dose drug-containing serum of musk and aconite,respectively.Cell viability was detected using the cell counting kit-8 method.Passage 3 human ventricular cardiomyocyte line AC16 was divided into four groups and treated with PBS,drug-free serum,and low-and high-dose drug-containing serum of musk-aconite,respectively,for 48 hours.Then,tumor necrosis factor α was added to the latter three groups for 4 hours to induce an inflammatory model.Western blot was used to detect the expression of nuclear factor κB inhibitor protein,phosphorylated nuclear factor κB inhibitor protein,nuclear factor κB p65,and nuclear factor κB p50 protein. RESULTS AND CONCLUSION:(1)A total of 268 prescriptions were identified from the 2020 edition of the Pharmacopoeia of the People's Republic of China.After removing duplicates and irrelevant prescriptions,88 prescriptions containing 36 Chinese medicinal herbs were selected.The most frequently used herbs were cinnamon,dried ginger,aconite,shallot,and musk.These herbs were predominantly pungent and warm in nature,targeting the heart,kidney,and liver meridians,with a treatment principle of Fangxiang Wentong method(aromatic warm and unblocking method).Association rule analysis identified four strong herb pairings,with high support for"musk-aconite,""cinnamon-dried ginger,""dried ginger-aconite,"and"shallot-trichosanthes"(confidence≥90%).Factor analysis extracted four common factors,reflecting the historical TCM treatment philosophy of Fangxiang Wentong method.The four core herb pairs might alleviate cardiac aging by modulating inflammatory signaling pathways,which were correlated with the relatively high proportion of inflammatory markers in cardiac aging.Molecular docking of the"musk-aconite"pair revealed good binding ability and stability between key targets and their corresponding active ingredients.(2)Cell counting kit-8 assay results indicated that the"musk-aconite"pair exhibited high cellular compatibility with drug-containing serum.Western blot analysis revealed that compared with the control group,intervention with tumor necrosis factor α significantly induced phosphorylation of the nuclear factor κB inhibitor protein,leading to the release of the nuclear factor κB dimer(p65/p50)into the cell nucleus,thereby confirming the successful establishment of the inflammatory injury model.However,low-and high-dose musk-aconite drug pair-containing serum intervention significantly inhibited the upregulation of nuclear factor κB inhibitor protein and nuclear factor κB(p65/p50)in the cell nucleus induced by tumor necrosis factor α.To conclude,in traditional Chinese medicine,the treatment of cardiac aging primarily follows the principle of Fangxiang Wentong method,with the musk-aconite pair as the core.This herbal combination may slow the progression of cardiac aging by inhibiting inflammatory signals.

Abstract:BACKGROUND:Under hypertensive conditions,exosome communication between endothelial progenitor cells and brain endothelial cells is impaired,which may be a key mechanism leading to poor prognosis in patients with hypertensive stroke.Exercise rehabilitation is an important non-drug means of preventing and treating hypertensive stroke,but the specific mechanism is unclear. OBJECTIVE:To investigate the effect of high-intensity interval training on the function of exosomes from endothelial progenitor cells in spontaneously hypertensive rats and explore the possible mechanism. METHODS:Twenty-four male spontaneously hypertensive rats were randomly assigned into exercise group or control group.The animals in the exercise group underwent 6 weeks of high-intensity interval training,while those in the control group were kept quietly in cage.After the experiment,bone marrow endothelial progenitor cells were isolated and cultured,and exosomes were obtained to detect the expression of miR-27a using qPCR.After treating neuroblastoma N2a(Neuro-2a)with angiotensin Ⅱ and hypoxia,they were co-incubated with endothelial progenitor cell-derived exosomes and a miR-27a inhibitor.The cells were then divided into the following groups:a blank group(cultured in normoxic high-glucose DMEM medium),an injury group(treated with angiotensin Ⅱ combined with hypoxia),an injury+control exosome group,an injury+exercise exosome group,and an injury+exercise exosome+miR-27a inhibitor group.The following parameters were assessed:exosome uptake rate,cell viability,reactive oxygen species levels,and the expression levels of cytochrome C and NADPH oxidase 4 protein. RESULTS AND CONCLUSION:(1)Exercise could increase the uptake efficiency of endothelial progenitor cell exosomes by Neuro-2a cells.(2)Exercise up-regulated the expression of miR-27a in endothelial progenitor cell exosomes and Neuro-2a cells(P<0.05).(3)Exercise-induced endothelial progenitor cell exosomes improved the survival of Neuro-2a cells(P<0.05),reduced the level of reactive oxygen species(P<0.05)and down-regulated the protein expression of cytochrome C and NADPH oxidase 4(P<0.05),while the above effects were weakened after administration of miR-27a inhibitor(P<0.05).To conclude,high-intensity interval training-induced exosomes from endothelial progenitor cells in spontaneously hypertensive rats alleviate the oxidative stress of damaged neurons through the miR-27a pathway.

Abstract:BACKGROUND:Cystic degeneration is a critical pathological feature of steroid-induced osteonecrosis of the femoral head and exhibits unusual reparative responses.Studies have shown that immune cell infiltration and upregulation of inflammatory cytokines in cystic areas may be closely related to bone regeneration during repair.Therefore,investigating the role of the immune microenvironment in cystic lesion repair has important clinical significance. OBJECTIVE:To explore the immune microenvironment in cystic degeneration of steroid-induced osteonecrosis of the femoral head based on single-cell sequencing and to elucidate the role of inflammation in the repair process as well as to validate the effects of glucocorticoids on macrophage polarization and inflammatory responses via in vitro cellular experiments. METHODS:(1)Single-cell RNA sequencing was used to analyze the immune microenvironment of cystic regions in steroid-induced osteonecrosis of the femoral head.Pathologic observation of cystic degeneration specimens was also performed.(2)In addition,an in vitro cell model was established.In vitro cultured RAW 264.7 macrophages were divided into control group(normal culture of RAW cells),lipopolysaccharide group(lipopolysaccharide intervention for 24 hours),and dexamethasone+lipopolysaccharide group(dexamethasone pretreatment for 72 hours and lipopolysaccharide intervention for 24 hours).RT-qPCR was used to detect the expression of cellular M1 polarization markers(inducible nitric oxide synthase,tumor necrosis factor α,and interleukin 6)and M2 polarization markers(arginase 1 and interleukin 10).ELISA was used to detect the levels of inflammatory factors in the cell culture supernatants.Flow cytometric assay was performed to detect the levels of cellular reactive oxygen species. RESULTS AND CONCLUSION:(1)Single-cell sequencing results revealed a diverse infiltration of immune cells in the cystic region,with significant upregulation of inflammatory cytokines such as tumor necrosis factor α and interleukin 6.(2)Observation of the pathologic specimen showed the presence of multiple inflammatory cell infiltrates in the cystic degeneration.(3)In vitro experiments demonstrated that combined treatment with dexamethasone and lipopolysaccharide significantly induced M1 polarization of RAW 264.7 macrophages,inhibited the expression of anti-inflammatory M2 phenotype and promoted the generation of reactive oxygen species,thereby enhancing the local inflammatory response(significant upregulation of tumor necrosis factor α,interleukin 6 and interleukin 1β).Overall,the immune microenvironment in the cystic regions of steroid-induced osteonecrosis of the femoral head plays a critical role in bone repair.Immune cell infiltration and inflammatory cytokine expression may exert dual effects during the reparative process.Excessive immune activation may lead to repair failure.Thus,modulating immune responses—especially balancing M1/M2 macrophage polarization—may serve as a potential therapeutic strategy for enhancing cystic repair in steroid-induced osteonecrosis of the femoral head.

Abstract:BACKGROUND:Developmental dysplasia of the hip is the most common skeletal deformity in children and is an important risk factor for osteoarthritis in young adulthood.Severe cases require hip replacement,which seriously affects the quality of life of patients. OBJECTIVE:To investigate the expression of secreted modular calcium-binding protein 2(SMOC2)in the cartilage acetabular roof of rats with developmental dysplasia of the hip and its effect on chondrocyte autophagy. METHODS:(1)Newborn Sprague-Dawley rats were randomly divided into disease group and control group(normal rats).A developmental dysplasia of the hip model was established in rats of the disease group.At 2 weeks of age,the expression of SMOC2 as well in the cartilage tissues of the acetabular top wall was detected by real-time quantitative polymerase chain reaction and Western blot.Autophagy-related protein expression was assessed via Western blot.(2)Chondrocytes were isolated from rat acetabular tissue and divided into three groups:Sh-NC,Sh-SMOC2-1,and Sh-SMOC2-2 groups.The expression of autophagy related proteins was detected by Western blot,and the number of autophagosomes was observed using a transmission electron microscope.(3)Rat acetabular tissue chondrocytes were extracted,and the cells were divided into three groups:Sh-NC,Sh-SMOC2,and Sh-SMOC2+740 Y-P(phosphatidylinositol 3-kinase activator)groups.The expression of autophagy-related proteins was detected using western blot to investigate whether SMOC2 regulates autophagy-related proteins of chondrocytes through the PI3K/AKT pathway. RESULTS AND CONCLUSION:(1)Compared with the control group,there was a decrease in the expression of SMOC2,a decrease in the expression of autophagy-associated protein P62,an increase in the expression of Beclin,and no difference in the expression of microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ in cartilage tissues of the rat acetabular top wall in the disease group.(2)Compared with the Sh-NC group,P62 expression was decreased,microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ ratio was increased,and Beclin1 expression was decreased in chondrocytes of the Sh-SMOC2-1 and Sh-SMOC2-2 groups.(3)Compared with the Sh-SMOC2 group,P62 expression was elevated,microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ ratio was decreased,and there was no difference in Beclin1 expression in chondrocytes of the Sh-SMOC2+740 Y-P group.In conclusion,autophagy is enhanced in acetabular chondrocytes of 2-week-old rats with developmental dysplasia of the hip,and SMOC2 can regulate autophagy in rat chondrocytes through the phosphatidylinositol 3-kinase/AKT signaling pathway.

Abstract:BACKGROUND:As a typical representative of neurodegenerative diseases,Parkinson's disease manifests with an intricate web of pathogenic attributes.In the field of programmed cell death,several novel molecular mechanisms have been unveiled in recent years,including but not limited to mitochondrial selective autophagy,iron-dependent cell death,copper ion-mediated apoptosis,and cell damage patterns caused by aberrant disulfide bonds.Studies have indicated that these pathological changes not only contribute to the development of typical clinical symptoms in Parkinson's disease but also exhibit significant correlations with disease progression.From this,it becomes apparent through scholarly works that these mechanisms exert pivotal roles in neuronal dysfunction manifestation.However,uncharted scientific inquiries regarding detailed interactions at the molecular level among these interconnected pathways need to be solved urgently. OBJECTIVE:To review the molecular mechanisms of novel forms of programmed cell death(mitophagy,ferroptosis,cuproptosis,and disulfidptosis)and the roles of their interactions and synergistic effects in Parkinson's disease,and to explore the potential impact and therapeutic prospects of these novel forms of programmed cell death in the pathogenesis of Parkinson's disease. METHODS:The first author conducted a computer-based search of literature indexed in the CNKI and PubMed databases from 2015 to 2025 using the keywords of"Parkinson's disease,mitophagy,ferroptosis,disulfidptosis,cuproptosis,apoptosis,necroptosis,pyroptosis"in Chinese and English,respectively.Literature that is inconsistent with the research content,outdated,or duplicative was excluded.A total of 1 368 relevant articles were retrieved,and 87 articles were ultimately included in the review analysis. RESULTS AND CONCLUSION:(1)The new types of programmed cell death,including mitophagy,ferroptosis,cuproptosis,and disulfidptosis,are essential regulatory pathways for maintaining normal cellular renewal and homeostasis,making them a currently hot research area in life sciences.(2)Mitophagy is a selective autophagic process that can clear damaged mitochondria,thereby maintaining mitochondrial homeostasis in neuronal cells during Parkinson's disease,and playing a crucial role in regulating neuronal health and function.(3)Ferroptosis is a novel form of programmed cell death characterized by iron accumulation and lipid peroxidation,which can regulate Parkinson's disease through various pathways,including cystine/glutamate,iron metabolism,and polyunsaturated fatty acids.(4)Cuproptosis,as a newly discovered mode of cell death,involves the role of copper chelators and heat shock proteins in regulating α-synuclein aggregation and cell survival,potentially providing new targets for the treatment of Parkinson's disease.(5)Disulfidptosis is associated with the aggregation of α-synuclein,which is considered an important feature of Parkinson's disease.Its specific role in Parkinson's disease requires further research.(6)The interactive and synergistic effects of various cell death mechanisms in Parkinson's disease(such as Fe/Cu synergistic regulation of Parkinson's disease)hold great potential for the future.

Abstract:OBJECTIVE:Through a network meta-analysis integrating direct and indirect evidence,this study compared the effects of different non-invasive brain stimulation techniques and parameters on gait and balance functions of patients with Parkinson's disease,and ranked the best intervention schemes. METHODS:A literature search of CNKI,WanFang,VIP,CBM,PubMed,Cochrane Library,Embase and Web of Science was conducted to screen the randomized controlled trials related to non-invasive brain stimulation for gait and balance disorders in patients with Parkinson's disease.The search time limit was up to June 16,2025.Data of the included studies were extracted for statistical processing using RevMan 5.4.1 software and Stata 17.0 software. RESULTS:(1)A total of 47 trials were conducted,involving 2 767 patients(1 399 in the trial group and 1 368 in the control group).(2)The results of traditional meta-analysis indicated that high-frequency repetitive transcranial magnetic stimulation could reduce the scores on the third part of the Unified Parkinson's Disease Rating Scale and the Freezing of Gait Questionnaire,shorten the Timed Up and Go Test time,increase step length,improve gait speed,and raise the Berg Balance Scale score,all of which were superior to conventional treatment(P<0.05).Transcranial direct current stimulation could lower the Freezing of Gait Questionnaire score,shorten the Timed Up and Go Test time,increase step length,improve gait speed,and raise the Berg Balance Scale score,all of which were superior to conventional treatment(P<0.05).Low-frequency repetitive transcranial magnetic stimulation could reduce the Unified Parkinson's Disease Rating Scale Ⅲ score,and there was a significant difference compared with conventional treatment(P<0.05).There was no statistically significant improvement in step frequency for all three methods(P>0.05);however,due to the small number of studies,further verification is required.(3)The results of the network Meta-analysis indicated that in terms of reducing the Unified Parkinson's Disease Rating Scale Ⅲ score:high-frequency repetitive transcranial magnetic stimulation targeting bilateral regions of the primary motor cortex and dorsolateral prefrontal cortex produced cumulative probability ranking is the highest(95.2％),followed by stimulation on the primary motor cortex(72.5％);in terms of shortening the Timed Up and Go Test time:high-frequency repetitive transcranial magnetic stimulation produced cumulative probability ranking was the highest on the dorsolateral prefrontal cortex(85.5％),followed by stimulation on the primary motor cortex(69.0％);in terms of improving gait speed:high-frequency repetitive transcranial magnetic stimulation produced cumulative probability ranking was the highest on the dorsolateral prefrontal cortex(92.5％),followed by stimulation on the primary motor cortex(76.7％);in terms of increasing the Berg Balance Scale score:high-frequency repetitive transcranial magnetic stimulation produced cumulative probability ranking was the highest on the primary motor cortex(79.9％),followed by transcranial direct current stimulation on the cerebellum(79.8％).(4)The GRADE evidence quality assessment results show that the evidence levels for the Unified Parkinson's Disease Rating Scale Ⅲ,Freezing of Gait Questionnaire score,Timed Up and Go Test time,and stride length were moderate,while those for gait speed,step frequency,and Berg Balance Scale score were low. CONCLUSION:Different types of non-invasive brain stimulation can all improve the gait and balance functions of patients with Parkinson's disease.High-frequency repetitive transcranial magnetic stimulation targeting the dorsolateral prefrontal cortex is superior to that targeting the primary motor cortex in improving gait function(moderate evidence),high-frequency repetitive transcranial magnetic stimulation targeting the primary motor cortex is superior to transcranial direct current stimulation targeting the cerebellum in improving balance function(low-level evidence).

Abstract:BACKGROUND:Existing studies have shown that extracorporeal shock wave intervention can effectively inhibit the formation of tissue fibrosis.However,there are limited experimental studies and summaries regarding the anti-fibrotic effects of extracorporeal shock wave,and no comprehensive review of the associated signaling pathways has been conducted. OBJECTIVE:To review current clinical or preclinical primary studies and summarize the applications of extracorporeal shock wave in fibrotic tissues and the relevant molecular mechanisms,thereby providing novel insights for the clinical treatment of fibrotic diseases. METHODS:The relevant literatures in PubMed,Web of Science,CNKI,WanFang and VIP databases were searched.The search time was from January 2014 to September 2024,and the search terms were"extracorporeal shockwave therapy,shock wave therapy,shock wave,fibrosis,fibroses"in English and"shock wave,extracorporeal shockwave therapy,fibrosis,anti-fibrosis"in Chinese.A combination of subject headings and free terms was used for the search.After screening and excluding studies based on predefined inclusion and exclusion criteria,a total of 67 articles were ultimately included for review and analysis. RESULTS AND CONCLUSION:(1)Extracorporeal shock wave intervention is beneficial for various fibrosis-related diseases,including scars,musculoskeletal disorders,and organ diseases.It can effectively reduce the degree of tissue fibrosis and improve clinical symptoms in patients.(2)Extracorporeal shock wave mainly influences the expression of fibrosis-related cytokines and extracellular matrix composition by modulating signaling pathways such as transforming growth factor-β1,mitogen-activated protein kinases,vascular endothelial growth factor,and inflammatory signaling pathways,thereby inhibiting excessive fibrosis formation.(3)The current review identifies a limited number of clinical studies with insufficient data to support robust conclusions.Additionally,due to the singularity of experimental results and significant differences in intervention subjects and extracorporeal shock wave intervention protocols across studies,there may be an impact on the clarification of the specific dose-response mechanisms and the overall molecular action system of extracorporeal shock wave in anti-fibrosis.To conclude,extracorporeal shock wave may serve as an effective therapeutic approach for the clinical treatment of fibrosis-related diseases in the future.