Kdm6a-CNN1 axis orchestrates epigenetic control of traumainduced spinal cord microvascular endothelial cell senescence to balance neuroinflammation for improved neurological repair

Cellular senescence assumes pivotal roles in various diseases through the secretion of proinflammatory factors.Despite extensive investigations into vascular senescence associated with aging and degenerative diseases,the molecular mechanisms governing microvascular endothelial cell senescence induced by traumatic stress,particularly its involvement in senescence-induced inflammation,remain insufficiently elucidated.In this study,we present a comprehensive demonstration and characterization of microvascular endothelial cell senescence induced by spinal cord injury(SCI).Lysine demethylase 6A(Kdm6a),commonly known as UTX,emerges as a crucial regulator of cell senescence in injured spinal cord microvascular endothelial cells(SCMECs).Upregulation of UTX induces senescence in SCMECs,leading to an amplified release of proinflammatory factors,specifically the senescenceassociated secretory phenotype(SASP)components,thereby modulating the inflammatory microenvironment.Conversely,the deletion of UTX in endothelial cells shields SCMECs against senescence,mitigates the release of proinflammatory SASP factors,and promotes neurological functional recovery after SCI.UTX forms an epigenetic regulatory axis by binding to calponin 1(CNN1),orchestrating trauma-induced SCMECs senescence and SASP secretion,thereby influencing neuroinflammation and neurological functional repair.Furthermore,local delivery of a senolytic drug reduces senescent SCMECs and suppresses proinflammatory SASP secretion,reinstating a local regenerative microenvironment and enhancing functional repair after SCI.In conclusion,targeting the UTX-CNN1 epigenetic axis to prevent trauma-induced SCMECs senescence holds the potential to inhibit SASP secretion,alleviate neuroinflammation,and provide a novel treatment strategy for SCI repair.

Leisure-time physical activity and all-cause mortality and cardiovascular disease in adults with type 2 diabetes:Cross-country comparison of cohort studies

Purpose:This study aimed to quantify the dose-response association and the minimal effective dose of leisure-time physical activity(PA)to prevent mortality and cardiovascular disease in adults with type 2 diabetes.Methods:Cross-country comparison of 2 prospective cohort studies including 14,913 and 17,457 population-based adults with type 2 diabetes from the UK and China.Baseline leisure-time PA was self-reported and categorized by metabolic equivalent hours per week(MET-h/week)according to World Health Organization recommendations:none,below recommendation(>0-7.49 MET-h/week);at recommended level(7.5-14.9 MET-h/week);above recommendation(>15 MET-h/week).Mortality and cardiovascular disease data were obtained from national registries.Results:During a median follow-up of 12.4 and 9.7 years,in the UK and China cohorts,repectively,higher levels of leisure-time PA were inversely associated with all-cause(1571 and 2351 events)and cardiovascular mortality(392 and 1060 events),mostly consistent with a linear dose-response relationship.PA below,at,and above recommendations,compared with no activity,yielded all-cause mortality hazard ratios of0.94(95%confidence interval(95%CI):0.79-1.12),0.90(95%CI:0.74-1.10),and 0.85(95%CI:0.70-1.02)in British adults and 0.87(95%CI:0.68-1.10),0.88(95%CI:0.74-1.03),and 0.77(95%CI:0.70-0.85)in Chinese adults.Associations with cardiovascular mortality were more pronounced in British adults(0.80(95%CI:0.58-1.11),0.75(95%CI:0.52-1.09),and 0.69(95%CI:0.48-0.97))but less pronounced in Chinese adults(1.06(95%CI:0.76-1.47),1.01(95%CI:0.80-1.28),and 0.79(95%CI:0.69-0.92)).PA at recommended levels was not associated with lower rates of major adverse cardiovascular events(2345 and 4458 events).Conclusion:Leisure-time PA at the recommended levels was not convincingly associated with lower mortality and had no association with risk of major adverse cardiovascular events in British or Chinese adults with type 2 diabetes.Leisure-time PA above current recommendations may be needed to prevent cardiovascular disease and premature mortality in adults with type 2 diabetes.

Current perspectives of viral hepatitis

Viral hepatitis represents a major danger to public health,and is a globally leading cause of death.The five liver-specific viruses:Hepatitis A virus,hepatitis B virus,hepatitis C virus,hepatitis D virus,and hepatitis E virus,each have their own unique epidemiology,structural biology,transmission,endemic patterns,risk of liver complications,and response to antiviral therapies.There remain few options for treatment,in spite of the increasing prevalence of viral-hepatitiscaused liver disease.Furthermore,chronic viral hepatitis is a leading worldwide cause of both liver-related morbidity and mortality,even though effective treatments are available that could reduce or prevent most patients’complications.In 2016,the World Health Organization released its plan to eliminate viral hepatitis as a public health threat by the year 2030,along with a discussion of current gaps and prospects for both regional and global eradication of viral hepatitis.Today,treatment is sufficiently able to prevent the disease from reaching advanced phases.However,future therapies must be extremely safe,and should ideally limit the period of treatment necessary.A better understanding of pathogenesis will prove beneficial in the development of potential treatment strategies targeting infections by viral hepatitis.This review aims to summarize the current state of knowledge on each type of viral hepatitis,together with major innovations.

Epidemiology of insertional and midportion Achilles tendinopathy in runners:A prospective cohort study

Background:Achilles tendinopathy(AT)is a common problem among runners.There is only limited evidence for risk factors for AT,and most studies have not defined the AT subcategories.No study has compared the incidence and risk factors between insertional AT and midportion AT,though they are considered distinct.This study aimed to assess incidence and risk factors of AT based on data from a large prospective cohort.The secondary aim was to explore differences in risk factors between insertional and midportion AT.Methods:Participants were recruited from among registered runners at registration for running events.Questionnaires were completed at baseline,1 month before the event,1 week before the event,and 1 month after the event.Information concerning demographics,training load,registered events,and running-related injuries were collected at baseline.The follow-up questionnaires collected information about new injuries.A pain map was used to diagnose midportion and insertional AT.The primary outcome was the incidence of AT.Multivariable logistic regression analysis was applied to identify risk factors for the onset.Results:We included 3379 participants with a mean follow-up of 20.4 weeks.The incidence of AT was 4.2%.The proportion of insertional AT was 27.7%and of midportion AT was 63.8%;the remaining proportion was a combined type of insertional and midportion AT.Men had a significantly higher incidence(5%,95%confidence interval(95%CI):4.1%-6.0%)than women(2.8%,95%CI:2.0%-3.8%).AT in the past12 months was the most predominant risk factor for new-onset AT(odds ratio(OR)=6.47,95%CI:4.27-9.81).This was similar for both subcategories of AT(insertional:OR=5.45,95%CI:2.51-11.81;midportion:OR=6.96,95%CI:4.24-11.40).Participants registering for an event with a distance of 10/10.55 km were less likely to develop a new-onset AT(OR=0.59,95%CI:0.36-0.97)or midportion AT(OR=0.47,95%CI:0.23-0.93).Higher age had a significant negative association with insertional AT(OR=0.97,95%CI:0.94-1.00).Conclusion:The incidence of new-onset AT among recreational runners was 4.2%.The proportion of insertional and midportion AT was 27.7%and 63.8%,respectively.AT in the past 12 months was the predominant risk factor for the onset of AT.Risk factors varied between insertional and midportion AT,but we could not identify clinically relevant differences between the 2 subtypes.

Device-measured physical activity and sedentary time in the Nordic countries:A scoping review of population-based studies

Purpose:The purpose of this scoping review was to summarize and describe the methodology and results from population-based studies of physical activity and sedentary time measured with devices in the Nordic countries(Denmark,Finland,Iceland,Norway,and Sweden)and published in 2000 or later.Methods:A systematic search was carried out in PubMed and Web of Science in June 2023 using predefined search terms.Results:Fourteen unique research projects or surveillance studies were identified.Additionally,2 surveillance studies published by national agencies were included,resulting in a total of 16 studies for inclusion.National surveillance systems exist in Finland and Norway,with regular survey waves in school-aged children/adolescents and adults.In Denmark,recent nationally representative data have been collected in school children only.So far,Sweden has no regular national surveillance system using device-based data collection.No studies were found from Iceland.The first study was conducted in 2001 and the most recent in 2022,with most data collected from 2016 to date.Five studies included children/adole scents 6-18 years,no study included preschoolers.In total 11 studies included adults,of which 8 also covered older adults.No study focused specifically on older adults.The analytical sample size ranged from 205 to 27,890.Detailed methodology is presented,such as information on sampling strategy,device type and placement,wear protocols,and physical activity classification schemes.Levels of physical activity and sedentary time in children/adolescents,adults,and older adults across the Nordic countries are presented.Conclusion:A growing implementation of device-based population surveillance of physical activity and sedentary behavior in the Nordic countries has been identified.The variety of devices,placement,and data procedures both within and between the Nordic countries highlights the challenges when it comes to comparing study outcomes as well as the need for more standardized data collection.

Mechanism underlying the effects of exercise against type 2 diabetes:A review on research progress

Exercise has emerged as one of the important and effective non-drug therapies used for management of type 2 diabetes(T2D)in certain nations.The present report summarizes the latest findings from the research on the beneficial effect of exercise on T2D.The objectives were to provide references for the theoretical study and the clinical practice of exercise-based management of T2D,in addition to identify the limitations of the existing literature,thereby provide direction for future research in this field.

From London buses to activity trackers:A reflection of 70 years of physical activity research

In 1953,Morris et al.^(1)published their ground-breaking findings among London bus drivers and conductors,linking physical activity to a major chronic disease for the first time.This seminal paper marked the birth of a new field—physical activity epidemiology.^(1)Since then,the field has flourished,evidenced by mounting scientific publications.^(2)Our knowledge about the role of physical activity in disease prevention has expanded from coronary heart disease in Morris et al.’s^(1)initial study to dozens of additional diseases and conditions,such as diabetes,cancers,and dementia.^(3)More than 7 decades and thousands of publications later,we critically reflect on several aspects of the progress in physical activity epidemiology.

Evaluation of genetic response of mesenchymal stem cells to nanosecond pulsed electric fields by whole transcriptome sequencing

BACKGROUND Mesenchymal stem cells(MSCs)modulated by various exogenous signals have been applied extensively in regenerative medicine research.Notably,nanosecond pulsed electric fields(nsPEFs),characterized by short duration and high strength,significantly influence cell phenotypes and regulate MSCs differentiation via multiple pathways.Consequently,we used transcriptomics to study changes in messenger RNA(mRNA),long noncoding RNA(lncRNA),microRNA(miRNA),and circular RNA expression during nsPEFs application.AIM To explore gene expression profiles and potential transcriptional regulatory mechanisms in MSCs pretreated with nsPEFs.METHODS The impact of nsPEFs on the MSCs transcriptome was investigated through whole transcriptome sequencing.MSCs were pretreated with 5-pulse nsPEFs(100 ns at 10 kV/cm,1 Hz),followed by total RNA isolation.Each transcript was normalized by fragments per kilobase per million.Fold change and difference significance were applied to screen the differentially expressed genes(DEGs).Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to elucidate gene functions,complemented by quantitative polymerase chain reaction verification.RESULTS In total,263 DEGs were discovered,with 92 upregulated and 171 downregulated.DEGs were predominantly enriched in epithelial cell proliferation,osteoblast differentiation,mesenchymal cell differentiation,nuclear division,and wound healing.Regarding cellular components,DEGs are primarily involved in condensed chromosome,chromosomal region,actin cytoskeleton,and kinetochore.From aspect of molecular functions,DEGs are mainly involved in glycosaminoglycan binding,integrin binding,nuclear steroid receptor activity,cytoskeletal motor activity,and steroid binding.Quantitative real-time polymerase chain reaction confirmed targeted transcript regulation.CONCLUSION Our systematic investigation of the wide-ranging transcriptional pattern modulated by nsPEFs revealed the differential expression of 263 mRNAs,2 miRNAs,and 65 lncRNAs.Our study demonstrates that nsPEFs may affect stem cells through several signaling pathways,which are involved in vesicular transport,calcium ion transport,cytoskeleton,and cell differentiation.

Orthopedic revolution:The emerging role of nanotechnology

This review summarizes the latest progress in orthopedic nanotechnology,ex-plores innovative applications of nanofibers in tendon repair,and evaluates the potential of selenium and cerium oxide nanoparticles in osteoarthritis and osteo-blast differentiation.This review also describes the emerging applications of inje-ctable hydrogels in cartilage engineering,emphasizing the critical role of inter-disciplinary research and highlighting the challenges and future prospects of in-tegrating nanotechnology into orthopedic clinical practice.This comprehensive approach provides a holistic perspective on the transformative impact of nanote-chnology in orthopedics,offering valuable insights for future research and clinical applications.

Recent research progress from biological perspective on the mechanism of formation of osteoarthritis after anterior cruciate ligament injury

The anterior cruciate ligament(ACL)mainly plays a role in stabilizing the knee joint by limiting the forward translation of tibial force and rotational force at the tibial joint,and if this ligament is damaged,it will cause joint pain,limited mobility,knee instability,etc.According to related studies,the incidence of traumatic osteoarthritis(PTOA)after ACL injury is as high as 87%,although many studies have shown that patients with ACL injury are susceptible to PTOA,but the exact mechanism is currently unknown.This may be related to biological,structural,and mechanical factors caused by the ligament injury.Previous studies have shown that elevated inflammatory mediators in the joint cavity following ACL injury can lead to chondrocytes necrosis and degradation of the cartilage matrix.These potential biochemical mediators contribute to PTOA formation,and early intervention can reduce future episodes of PTOA.In recent years,many scholars have devoted themselves to studying the potential important factors and signaling pathways involved in the formation of osteoarthritis after ACL injury,and exploring its molecular mechanism,which has led to great progress in this field.This paper mainly studies and discusses the mechanism of osteoarthritis formation after ACL injury from the biological perspective.

Bioinformatics Analysis of the Biological Properties of Ewing Sarcoma

Purpose: Bioinformatics-based approach to screen and analyze differentially expressed genes associated with the biological characteristics of Ewing sarcoma. Means: The GSE17674 dataset was selected for analysis, obtained by data retrieval based on the GEO public database. The R language limma toolkit was used to screen DEmRNAs. After the data were normalized, the Metascape online analysis software and the R language clusterProfiler package were used to analyze the GO function and KEGG pathway enrichment of DEmRNAs lines, respectively. The string database was selected for PPI analysis, and the results were imported into Cytoscape software to derive the core modules and predicted core genes. The genes selected above were analyzed for tissue localization specificity. Results: Through the analysis of GSE17674, differentially expressed genes were screened out, and GO and KEGG analyses were performed on the differentially expressed genes. The GO functional enrichment analysis was mainly enriched in the process of muscle system, muscle contraction, myocyte development, contractile fibers, myogenic fibers, myofibers, myofibrillar segments, actin binding, structural composition of muscle, and actin filament binding. KEGG pathway analysis showed that the core pathways associated with the development of ES were the core genes for myocardial contraction, congestive cardiomyopathy, and hypertrophic cardiomyopathy. Five Hub genes were obtained based on Cytoscape prediction. Tissue localization specificity analysis of Hub genes was performed, and a total of 2 Hub genes with tissue specificity were screened;MYH6 was specifically expressed in cardiac cells and MYL1 was specifically expressed in skeletal muscle cells. Conclusions: The differential genes screened will help to understand the molecular mechanisms underlying the highly invasive and metastasis-prone biological characteristics of ES, as well as provide new ideas for clinical drug-targeted treatment of ES.

Research Progress of Radical Treatment of Helicobacter Pylori Based on Drug Sensitivity Test as First-Line Treatment

Helicobacter pylori (HP) infection is a global problem that affects about half of the world’s population and requires sufficient attention in clinical and scientific work. Due to differences in economic and medical conditions among countries around the world, there is currently no unified treatment plan for anti-HP. In China, empirical quadruple therapy is mainly used. With the abuse of antibiotics, many patients face the problem of secondary eradication after failure, and the resistance rate of HP is gradually increasing. After eradication failure, drug sensitivity cultivation is carried out to choose sensitive antibiotics for treatment. A new strategy is currently needed to address how to improve the eradication rate of HP during the first eradication. This article aims to discuss the first-line treatment plans and research progress for eradicating HP based on drug sensitivity testing before eradication. Compared with traditional empirical therapies, treatment based on drug sensitivity results can effectively improve the eradication rate of HP, and reduce drug resistance rates, and adverse reactions, among other benefits. .

Minimum 10-year follow-up outcomes of arthroscopic Bankart’s repair with metallic anchors:Reliable results with low redislocation rates

BACKGROUND With stiff competition from alternative albeit more expensive counterparts,it has become important to establish the applicability of metallic anchors for shoulder instability in the modern era.This can be accomplished,in part,by analysing long-term outcomes.AIM To analyse minimum 10-year outcomes from 30 patients following arthroscopic anterior stabilisation using metallic anchors.METHODS Prospectively collected data from arthroscopic Bankart repairs performed using metal anchors during 2007P-2010 were retrospectively analysed in this singlesurgeon study.Comprehensive data collection included historical and clinical findings,dislocation details,operative specifics,and follow-up radiological and clinical findings including shoulder scores.The primary outcomes were patientreported scores(Constant,American Shoulder and Elbow Surgeons[ASES],and Rowe scores)and pain and instability on a visual analogue scale(VAS).RESULTS A 3% recurrence rate of dislocation was noted at the final follow-up.Total constant scores at 10 years postoperatively measured between 76 and 100(mean 89)were significantly better than preoperative scores(mean 62.7).Congruous improvements were also noted in the Rowe and ASES scores and VAS at the 10-year review.CONCLUSION Reliable long-term outcomes with metallic anchors in surgery for shoulder instability can be expected.Our results provide additional evidence of their continued,cost-effective presence in the modern scenario.

Single-Arm Clinical Study of Combination Perindopril-Amlodipine Tablets in the Treatment of High-Altitude Hypertension

Objective:To evaluate the efficacy of combination perindopril/amlodipine tablets in patients with high-altitude hypertension who were previously unable to control their blood pressure with monotherapy.Methods:A total of 151 patients with high-altitude hypertension whose blood pressure remained inadequately controlled with previous monotherapy were enrolled in this study.All patients received an 8-week treatment with a combination of perindopril/amlodipine tablets,consisting of perindopril 10 mg/day and amlodipine 5 mg/day.Blood pressure measurements,including both diastolic and systolic pressures,were taken at baseline,and after 2,4,6,and 8 weeks of treatment.Results:After 8 weeks of treatment,there was a significant reduction in both average systolic and diastolic blood pressure compared to baseline(P<0.0001).Specifically,the average systolic blood pressure decreased by 24.45±13.75 mmHg,and the average diastolic blood pressure decreased by 13.37±8.40 mmHg.The overall heart rate showed no significant changes during the treatment period.Conclusion:A combination of perindopril/amlodipine tablets significantly improved blood pressure control in patients with high-altitude hypertension after 8 weeks of treatment.These results support the efficacy of combination perindopril/amlodipine as a viable treatment option for high-altitude hypertension.

Comparative study of the clinical efficacy of all-inside and traditional techniques in anterior cruciate ligament reconstruction

BACKGROUND Many studies have focused on the femoral tunnel technique and fixation method,but few studies have involved the tibial tunnel technique and fixation method.The all-inside technique is one of the new techniques that has been described in recent years.All-inside anterior cruciate ligament(ACL)reconstruction is based on a tibial socket instead of a full tunnel.This method has many potential advantages.AIM To compare clinical outcomes of knee ACL autograft reconstruction using allinside quadrupled semitendinosus(AIST)and traditional hamstring tendon(TBT)techniques.METHODS From January 2017 to October 2019,the clinical data of 80 patients with ACL reconstruction were retrospectively analyzed,including 67 males and 13 females.The patients had an average age of 24.3±3.1 years(age range:18-33 years).The AIST technique was used in 42 patients and the TBT technique was used in 38 patients.The time between operation and injury,operative duration,postoperative visual analogue scale(VAS)score and knee functional recovery were recorded and compared between the two groups.The International Knee Documentation Committee(IKDC)and Lysholm scoring system were used to comprehensively evaluate clinical efficacy.RESULTS Eighty patients were followed for 24-36 mo,with an average follow-up duration of 27.5±1.8 mo.There were no significant differences in the time between surgery and injury,operative duration,IKDC and Lysholm scores of the affected knee at the last follow-up evaluation between the two groups.There were significant differences in VAS scores 1 d,3 d,7 d,2 wk and 1 mo after surgery(P<0.05).There was no significant difference in VAS score at 3 mo,6 mo and 1 year after operation.CONCLUSION The efficacy of the AIST ACL reconstruction technique was comparable to the TBT technique,but the postoperative pain was less with the AIST technique.Thus,the AIST technique is an ideal treatment choice for ACL reconstruction.

Research progress on preparation of lateral femoral tunnel and graft fixation in anterior cruciate ligament reconstruction

Anterior cruciate ligament(ACL)injury is one of the most common types of sports injuries.People’s need to participate in sports and desire for a high quality of life promotes the continuous development of ACL reconstruction technology.Arthroscopic ACL reconstruction has been recognized as an effective method for the treatment of ACL injuries.This review analyses and summarizes the advantages and limitations of each surgical procedure for arthroscopic ACL reconstruction reported in the relevant literature so as to promote the future development of more relevant techniques.

Long head of biceps tendon transposition for massive and irreparable rotator cuff tears:A systematic review and meta-analysis

BACKGROUND Superior capsular reconstruction(SCR)with long head of biceps tendon(LHBT)transposition was developed to massive and irreparable rotator cuff tears(MIRCTs);however,the outcomes of this technique remain unclear.AIM To perform a systematic review of biomechanical outcomes and a meta-analysis of clinical outcomes after LHBT transposition for MIRCTs.METHODS We performed a systematic electronic database search on PubMed,EMBASE,and Cochrane Library.Studies of SCR with LHBT transposition were included according to the inclusion and exclusion criteria.Biomechanical studies were assessed for main results and conclusions.Included clinical studies were evaluated for quality of methodology.Data including study characteristics,cohort demographics,and outcomes were extracted.A meta-analysis was conducted of the clinical outcomes.RESULTS According to our inclusion and exclusion criteria,a total of six biomechanical studies were identified and reported an overall improvement in subacromial contact pressures and prevention of superior humeral migration without limiting range of motion(ROM)after LHBT transposition for MIRCTs.A total of five clinical studies were included in the meta-analysis of LHBT transposition outcomes,consisting of 253 patients.The results indicated that compared to other surgical methods for MIRCTs,LHBT transposition had advantages of more significant improvement in ROM(forward flexion mean difference[MD]=6.54,95%confidence interval[CI]:3.07-10.01;external rotation[MD=5.15,95%CI:1.59-8.17];the acromiohumeral distance[AHD][MD=0.90,95%CI:0.21-1.59])and reducing retear rate(odds ratio=0.27,95%CI:0.15-0.48).No significant difference in American Shoulder and Elbow Surgeons score,visual analogue scale score,and University of California at Los Angles score was demonstrated between these two groups for MIRCTs.CONCLUSION In general,SCR with LHBT transposition was a reliable and economical technique for treating MIRCTs,both in terms of biomechanical and clinical outcomes,with comparable clinical outcomes,improved ROM,AHD,and reduced the retear rates compared to conventional SCR and other established techniques.More high-quality randomized controlled studies on the long-term outcomes of SCR with LHBT transposition are required to further assess.

High doses of dextromethorphan induced shock and convulsions in a 19-year-old female:A case report

BACKGROUND Dextromethorphan is a prevalent antitussive agent that can be easily obtained as an over-the-counter medication.There has been a growing number of reported cases of toxicity in recent years.Generally,there are numerous instances of mild symptoms,with only a limited number of reports of severe cases necessitating intensive care.We presented the case of a female who ingested 111 tablets of dextromethorphan,leading to shock and convulsions and requiring intensive care that ultimately saved her life.CASE SUMMARY A 19-year-old female was admitted to our hospital via ambulance,having overdosed on 111 tablets of dextromethorphan(15 mg)obtained through an online importer in a suicide attempt.The patient had a history of drug abuse and multiple self-inflicted injuries.At the time of admission,she exhibited symptoms of shock and altered consciousness.However,upon arrival at the hospital,the patient experienced recurrent generalized clonic convulsions and status epilepticus,necessitating tracheal intubation.The convulsions were determined to have been caused by decreased cerebral perfusion pressure secondary to shock,and noradrenaline was administered as a vasopressor.Gastric lavage and activated charcoal were also administered after intubation.Through systemic management in the intensive care unit,the patient’s condition stabilized,and the need for vasopressors ceased.The patient regained consciousness and was extubated.The patient was subsequently transferred to a psychiatric facility,as suicidal ideation persisted.CONCLUSION We report the first case of shock caused by an overdose of dextromethorphan.

Highly specific characterization and discrimination of monosodium urate crystals in gouty arthritis based on aggregation-induced emission luminogens

Existing technologies used to detect monosodium urate(MSU)crystals for gout diagnosis are not ideal due to their low sensitivity and complexity of operation.The purpose of this study was to explore whether aggregation-induced emission luminogens(AIEgens)can be used for highly specific imaging of MSU crystals to assist in the diagnosis of gout.First,we developed a series of luminogens(i.e.,tetraphenyl ethylene(TPE)-NH_(2),TPE-2NH_(2),TPE-4NH_(2),TPE-COOH,TPE-2COOH,TPE-4COOH,and TPE-Ketoalkyne),each of which was then evenly mixed with MSU crystals.Next,optimal fluorescence imaging of each of the luminogens was characterized by a confocal laser scanning microscope(CLSM).This approach was used for imaging standard samples of MSU,hydroxyapatite(HAP)crystals,and mixed samples with 1:1 mass ratio of MSU/HAP.We also imaged samples from mouse models of acute gouty arthritis,HAP deposition disease,and comorbidities of interest.Subsequently,CLSM imaging results were compared with those of compensated polarized light microscopy,and we assessed the biosafety of TPE-Ketoalkyne in the RAW264.7 cell line.Finally,CLSM time series and three-dimensional imaging were performed on MSU crystal samples from human gouty synovial fluid and tophi.As a promising candidate for MSU crystal labeling,TPE-Ketoalkyne was found to detect MSU crystals accurately and rapidly in standard samples,animal samples,and human samples,and could precisely distinguish gout from HAP deposition disease.This work demonstrates that TPE-Ketoalkyne is suitable for highly specific and timely imaging of MSU crystals in gouty arthritis and may facilitate future research on MSU crystal-related diseases.

Bone marrow mesenchymal stem cell-derived exosomal microRNAs target PI3K/Akt signaling pathway to promote the activation of fibroblasts

BACKGROUND Fibroblast plays a major role in tendon-bone healing.Exosomes derived from bone marrow mesenchymal stem cells(BMSCs)can activate fibroblasts and promote tendon-bone healing via the contained microRNAs(miRNAs).However,the underlying mechanism is not comprehensively understood.Herein,this study aimed to identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets,and to verify their effects as well as mechanisms on fibroblasts.AIM To identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets and verify their effects as well as mechanisms on fibroblasts.METHODS BMSC-derived exosomal miRNAs data(GSE71241,GSE153752,and GSE85341)were downloaded from the Gene Expression Omnibus(GEO)database.The candidate miRNAs were obtained by the intersection of three data sets.TargetScan was used to predict potential target genes for the candidate miRNAs.Functional and pathway analyses were conducted using the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases,respectively,by processing data with the Metascape.Highly interconnected genes in the protein-protein interaction(PPI)network were analyzed using Cytoscape software.Bromodeoxyuridine,wound healing assay,collagen contraction assay and the expression of COL I andα-smooth muscle actin positive were applied to investigate the cell proliferation,migration and collagen synthesis.Quantitative real-time reverse transcription polymerase chain reaction was applied to determine the cell fibroblastic,tenogenic,and chondrogenic potential.RESULTS Bioinformatics analyses found two BMSC-derived exosomal miRNAs,has-miR-144-3p and hasmiR-23b-3p,were overlapped in three GSE datasets.PPI network analysis and functional enrichment analyses in the GO and KEGG databases indicated that both miRNAs regulated the PI3K/Akt signaling pathway by targeting phosphatase and tensin homolog(PTEN).In vitro experiments confirmed that miR-144-3p and miR-23b-3p stimulated proliferation,migration and collagen synthesis of NIH3T3 fibroblasts.Interfering with PTEN affected the phosphorylation of Akt and thus activated fibroblasts.Inhibition of PTEN also promoted the fibroblastic,tenogenic,and chondrogenic potential of NIH3T3 fibroblasts.CONCLUSION BMSC-derived exosomes promote fibroblast activation possibly through the PTEN and PI3K/Akt signaling pathways,which may serve as potential targets to further promote tendon-bone healing.