Abstract:BACKGROUND:Adipocyte differentiation is a complex biological process involving the transformation of preadipocytes into mature adipocytes.This process plays a key role in the development and progression of obesity and related metabolic diseases.In recent years,the complement C1q/tumor necrosis factor-related protein family,as a new member of the adipokine family,has become a research hotspot in the field of metabolic regulation. OBJECTIVE:To investigate the effect of complement 1q/tumor necrosis factor-related protein 4 on the differentiation of 3T3-L1 preadipocytes and its underlying mechanism. METHODS:The CCK-8 assay was used to determine the effects of different concentrations of complement 1q/tumor necrosis factor-related protein 4 on the viability of 3T3-L1 preadipocytes,and a safe concentration was selected for intervention.3T3-L1 preadipocytes were divided into a control group,an inducer group,and a complement 1q/tumor necrosis factor-related protein 4 group.3T3-L1 preadipocytes were first contact-inhibited for 2 days,followed by adipogenic inducer and complement 1q/tumor necrosis factor-related protein 4 treatment.On day 10 of differentiation,lipid droplet formation was observed using Oil Red O staining.The mRNA and protein expression levels of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4 were determined by RT-qPCR and western blot assay.Intracellular triglyceride and total cholesterol levels were measured using triglyceride and total cholesterol detection kits. RESULTS AND CONCLUSION:The safe maximum concentration of complement 1q/tumor necrosis factor-related protein 4 without toxicity to 3T3-L1 preadipocytes was 1 000 ng/mL.Compared with the control group,the inducer group showed increased intracellular lipid droplet formation,elevated triglyceride and total cholesterol levels(P<0.05),and significantly upregulated mRNA and protein expression levels of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4(P<0.05).Compared with the inducer group,the complement 1q/tumor necrosis factor-related protein 4 group showed reduced intracellular lipid droplet formation,lower triglyceride and total cholesterol levels(P<0.05).The mRNA and protein expression levels of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4 were significantly downregulated(P<0.05).These results suggest that complement 1q/tumor necrosis factor-related protein 4 inhibits the differentiation of preadipocytes into mature adipocytes,and its mechanism of action may be related to downregulating the expression of CCAAT/enhancer binding protein α,peroxisome proliferator-activated receptor γ,and fatty acid binding protein 4.

Abstract:BACKGROUND:The anterior transpedicular screw fixation system can achieve adequate decompression and strong fixation in a single anterior surgery.However,due to its unique anatomical structure,the risk of screw placement is relatively high.Previous domestic and international scholars have confirmed the feasibility of this approach through anatomical measurements,but there are some drawbacks such as relatively small sample sizes,difficulties in locating the pedicle axis,and limitations in measurement methods. OBJECTIVE:To perform morphological measurements of the adult cervical spine based on imaging to provide anatomical guidance for anterior transpedicular pedicle screw fixation. METHODS:3D CT scan data of 50 adult cervical vertebrae were imported into the Mimics system for 3D reconstruction.Morphological data were measured,including pedicle axis distance,pedicle width,pedicle height,pedicle horizontal axial angle,pedicle sagittal angle,distance from the entry point in the transverse plane,distance to the entry point in the sagittal plane,axial vertebral length and axial pedicle length. RESULTS AND CONCLUSION:(1)Positioning of nail entry points:C3 and C4 were located on the opposite side of the median sagittal plane of the vertebral body,with distances from the median sagittal line of approximately 2.060 mm and 2.310 mm.C5 could be located on the same side of the median sagittal line as or opposite to the median sagittal line,with an average value of approximately 1.224 mm.C6-C7 were located on the same side of the body,with distances from the center of the vertebral body of 1.132 mm and 2.538 mm;the distances of C3-C7 from the vertebral body of the upper endplates increasd gradually,with average values ranging from 2.362-7.350 mm.(2)In the direction of the entry point,the cross-sectional pinch angles:C3-4 gradually increased to 46.32°-47.36°,and C5-7 gradually decreased from 44.03° to 37.80°;the sagittal pinch angles:C3-C4 needed to be shifted caudally,with the angles of 95.75° and 100.93°;C5-C7 angles needed to be shifted toward the cephalad,with angles of 104.38°,110.34°,and 104.86°,respectively.(3)There was no statistical significance in the positioning and orientation of the entry point between genders and sides(P>0.05).For screw selection,except for individual patients with abnormal cervical vertebral development,the vast majority of the lower cervical pedicle screws were selected to be at least 30-mm-long,4.0-mm-diameter screws for males,and at least 28-mm-long,3.5-mm-diameter screws for females for anterior transpedicular pedicle screw fixation,which could ensure their safety and reliability while to achieve bicortical screws.(4)Morphological measurements confirm that anterior transpedicular screw fixation of the lower cervical spine is feasible,but a personalized fixation plan should be developed based on individual needs.

Abstract:BACKGROUND:Neuropathic pain has a complex pathogenesis and limited clinical intervention outcomes.In recent years,exosomes have gradually emerged as a focal point in the study of neuropathic pain due to their unique intercellular communication functions and molecular delivery capabilities. OBJECTIVE:To systematically review research progress on exosomes in the field of neuropathic pain using bibliometric methods,summarize the knowledge framework and research hotspots,and provide theoretical foundations and translational strategies for advancing this field. METHODS:Based on the Web of Science Core Collection(WOSCC)database,relevant literature on exosomes and neuropathic pain was retrieved over the time span from January 1,2012,to March 31,2025.Keyword co-occurrence analysis,clustering analysis,outbreak detection,and collaboration network visualization were performed using VOSviewer and CiteSpace software. RESULTS AND CONCLUSION:A total of 313 articles were included in this study.Among 42 countries or regions,the United States and China made significant contributions.Shanghai Jiao Tong University and Nantong University were the research institutions with the highest number of publications.The journals with the most publications and the highest number of citations were Neural Regeneration Research and International Journal of Molecular Sciences,respectively.The study identified 382 authors,with Zhang Zheng Gang having the highest number of publications and Zhang Yi having the highest number of citations.Keywords of particular interest included"nerve regeneration,""neuroinflammation,""Schwann cell,""regenerative medicine,"and"spinal cord injury,"all of which are key areas for future research.Bibliometric analysis revealed research trends of exosomes in the field of neuropathic pain.Exosomes hold great promise in elucidating the underlying mechanisms and developing novel precision therapies for neuropathic pain.Future studies may combine multi-omics and engineered exosome platforms to advance the development of this field.

Abstract:BACKGROUND:Non-infectious poor wound healing following lumbar surgery is a significant clinical complication that prolongs hospitalization and increases the risk of reoperation.However,its predictive indicators remain unclear.Based on the hypothesis that paraspinal muscle degeneration may impede tissue repair by altering the local microenvironment,this study aimed to investigate the predictive value of preoperative paraspinal muscle fatty infiltration for non-infectious poor wound healing and its association with osteoporosis. OBJECTIVE:To quantify the degree of paraspinal muscle fatty infiltration using preoperative MRI and evaluate its predictive value for non-infectious poor wound healing after lumbar surgery. METHODS:A retrospective analysis was conducted on medical records of 4 368 patients who underwent traditional open posterior lumbar surgery at Third Affiliated Hospital of Guangzhou University of Chinese Medicine between 2019 and 2024.We screened 190 patients with a postoperative hospital stay of 15 days or longer.Based on postoperative wound healing and infection indicators,patients with non-infectious poor healing were identified from these 190 patients and placed in a poor healing group(41 patients).The remaining 4 178 surgical patients were screened and their medical records and blood tests were reviewed.Patients with good healing were selected and placed in a good healing group(40 patients).The non-infectious poor healing group was further divided into an osteoporosis group and a non-osteoporosis group.Preoperative lumbar MRI images were analyzed using ImageJ software to measure and calculate the cross-sectional area of the psoas major muscle and the percentage of fat infiltration in the erector spinae and multifidus muscles. RESULTS AND CONCLUSION:(1)No significant differences were observed in baseline characteristics such as gender,age,or diabetes status between the poor healing group and the good healing group(P>0.05).(2)There were significant differences in the functional cross-sectional area and fat infiltration percentage of the psoas major,erector spinae,and multifidus muscles between the poor healing group and the good healing group(P<0.05).(3)Logistic regression analysis showed that the fat infiltration percentage was an independent risk factor for poor wound healing.(4)Receiver operating characteristic curve analysis showed that the fat infiltration percentage had a high predictive value for poor wound healing(area under the curve>0.7).(5)One-way analysis of variance suggested that osteoporosis may be a risk factor for the degree of fatty infiltration of the L4 multifidus muscle(P<0.05).(6)The results indicate that the percentage of fatty infiltration of the paraspinal muscles is an important indicator for predicting poor wound healing in patients undergoing traditional posterior open lumbar spine surgery,providing a clinical reference.Osteoporosis has also been shown to be a risk factor for the degree of fatty infiltration of the L4 multifidus muscle.However,due to the small sample size in this subgroup,further clinical trials are needed to confirm whether osteoporosis affects non-infectious poor wound healing after lumbar spine surgery.

Abstract:BACKGROUND:The scarcity of bone trabecular structures caused by osteoporosis is not enough to maintain screw holding power,which often leads to the failure of internal fixation surgery.The screw holding power is often increased by increasing the diameter and length of screws,changing the surface coating of screws,and reinforcing the nail track with bone cement.The cement reinforced cortical bone track nailing technique and the modified cortical bone track nailing technique using a new type of variable diameter screw have been proven to have good fixation effects,and now the related mechanical properties of the two need to be analyzed and compared. OBJECTIVE:Finite element analysis was used to compare the mechanical properties of bone cement-strengthened cortical bone trajectory nailing technology,cortical bone trajectory nailing technology,and modified cortical bone trajectory nailing technology using a new variable-diameter total cortical bone thread screw in lumbar spine internal fixation surgery. METHODS:Based on the CT scan data processing of three osteoporotic vertebrae,the L4 lumbar spine model was constructed,and the innovative variable-diameter all-cortical bone screw was applied in the modified cortical bone nailing technique,with the screw having a total length of 45 mm and a variable diameter of 5.5-4.3 mm as the modified cortical bone trajectory screw group.It was compared with the standard cortical bone trajectory screw group(5.5 mm in diameter and 40 mm in length)and the bone cement-augmented cortical bone trajectory screw group(5.5 mm in diameter,40 mm in length,and 1 mm in diameter in the center opening).The fixation strength of the screws in each group was compared by measuring the axial withdrawal force,screw stability(upper,lower,left,and right load displacement ratio),and vertebral body mobility. RESULTS AND CONCLUSION:(1)Axial pullout strength:The bone cement-augmented cortical bone trajectory screw group showed greater axial pullout strength than the modified cortical bone trajectory screw group(P=0.024 6).Additionally,both the bone cement-augmented cortical bone trajectory screw group and the modified cortical bone trajectory screw group exhibited significantly higher axial pullout strength compared with the standard cortical bone trajectory screw group(P=0.000 1 and P=0.002 64,respectively).(2)Screw stability:Under inferior loading conditions,the load-displacement ratio of the screws was highest in the bone cement-augmented cortical bone trajectory screw group,followed by the standard cortical bone trajectory screw group and the modified cortical bone trajectory screw group(all P<0.05).The modified cortical bone trajectory screw group showed a higher stability compared with the standard cortical bone trajectory screw group(P<0.05).(3)Vertebral mobility:Under five different loading conditions,the bone cement-augmented cortical bone trajectory screw group exhibited less vertebral mobility than the modified cortical bone trajectory screw group,and no significant difference was detected(P>0.05).Furthermore,both the bone cement-augmented cortical bone trajectory screw group and the modified cortical bone trajectory screw group showed less vertebral mobility compared with the standard cortical bone trajectory screw group.(4)There were no statistically significant differences between the bone cement-augmented cortical bone trajectory screw group and the modified cortical bone trajectory screw group in terms of load-displacement ratio and lumbar mobility(P>0.05).(5)This suggests that the bone cement-augmented cortical bone trajectory screw technique offers superior biomechanical performance compared with the novel variable-diameter full-cortical bone screw.Additionally,the bone cement-augmented cortical bone trajectory screw technique may be a more suitable choice for internal fixation surgery in clinical patients with osteoporosis.

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:Hyperuricemia can induce kidney damage and activate inflammatory signaling pathways,leading to glomerular hypertrophy and the deterioration of renal function.Exercise,as a non-pharmacological treatment method,can regulate the expression of proteins related to uric acid excretion.However,the regulatory mechanism of exercise on hyperuricemia remains unclear. OBJECTIVE:To explore the potential effect and molecular mechanism of moderate-intensity exercise on hyperuricemia. METHODS:(1)Using the Mendelian randomization analysis method,the moderate-intensity exercise datasets(ukb-a-508,ukb-b-4710)from the database developed by the MRC Integrative Epidemiology Unit at the University of Bristol were used as the exposure factors,and the serum uric acid level dataset(ebi-a-GCST90018977)was used as the outcome indicator.Meanwhile,the gout datasets(finn-b-GOUT,finn-b-M13GOUT)from the FinnGen,a genomics and personalized medicine research project in Finland,were included as another outcome indicator to explore the potential associations among moderate-intensity exercise,gout,and uric acid levels.(2)An animal experiment was conducted to explore the specific mechanism between moderate-intensity exercise and hyperuricemia.Four groups were set up:blank control group,moderate-intensity exercise+blank group,hyperuricemia model group,and moderate-intensity exercise+hyperuricemia model group.After 8 weeks of moderate-intensity treadmill training in the experimental C57BL/6 mice,biochemical indicators such as serum uric acid,creatinine,and blood urea nitrogen were detected.Hematoxylin-eosin staining was used to observe the pathological changes in kidney tissues.Additionally,qRT-PCR and western blot techniques were employed to analyze the gene and protein expression levels of Nod-like receptor family pyrin domain containing-3,Caspase-1,interleukin-1β,interleukin-6,and interleukin-11. RESULTS AND CONCLUSION:(1)The results of Mendelian randomization showed that there was a significant negative correlation between moderate-intensity exercise and both gout and serum uric acid level(P<0.05).(2)Compared with the blank control group,the levels of serum uric acid,creatinine,and blood urea nitrogen in the hyperuricemia model group increased significantly(P<0.05).(3)Compared with the blank control group,the hyperuricemia model group had pathological changes such as glomerular hypertrophy,vacuolar degeneration of renal tubular epithelial cells,and infiltration of inflammatory cells.(4)Compared with the blank control group,the mRNA expression levels of Nod-like receptor family pyrin domain containing-3,interleukin-1β,and interleukin-18 and the expression levels of Nod-like receptor family pyrin domain containing-3,Caspase-1,interleukin-1β,interleukin-6,and interleukin-11 in the hyperuricemia model group were significantly upregulated(P<0.05).(5)After moderate-intensity exercise intervention,the above detection indices in the moderate-intensity exercise+hyperuricemia model group decreased significantly(P<0.05),and the morphology of the renal tissue was improved.These findings suggest that moderate-intensity exercise can reduce the uric acid level and renal injury in hyperuricemia mice.In addition,moderate-intensity exercise can inhibit the activation of the Nod-like receptor family pyrin domain containing-3 signaling pathway and alleviate kidney inflammation.