Abstract:BACKGROUND:The anatomical morphology of the distal femur and proximal tibia has an important influence on the movement of the tibiofemoral joint,and its role in anterior cruciate ligament injury has been well described.However,the research on the anatomical risk factors of posterior cruciate ligament tibial avulsion fracture is still limited,and no unified consensus has been formed. OBJECTIVE:To investigate the anatomical risk factors for posterior cruciate ligament tibial avulsion fracture. METHODS:The medical records of 53 patients who underwent surgical treatment for posterior cruciate ligament tibial avulsion fracture from March 2021 to September 2024 were retrospectively collected as the posterior cruciate ligament avulsion group(32 males and 21 females),and the data of 53 subjects without posterior cruciate ligament injury in the same period were included as the posterior cruciate ligament normal group(24 males and 29 females).The intercondylar notch width,femoral condyle width,intercondylar notch width index,intercondylar notch height,intercondylar notch shape index,intercondylar notch angle,Blumensaat's line inclination angle,medial tibial posterior slope,and lateral tibial posterior slope were measured and calculated in Magnetic Resonance Imaging to analyze the differences in the anatomical data of the two groups.Binary logistic regression analysis was used to determine the independent risk factors and to establish a risk factor model by receiver operating characteristic curve. RESULTS AND CONCLUSION:(1)The intercondylar notch width index,intercondylar notch shape index,intercondylar notch angle,and medial tibial posterior slope of the posterior cruciate ligament tibial avulsion group were smaller than those of the posterior cruciate ligament normal group,and the differences were statistically significant(P<0.05).The data of the two groups were not statistically different in the intercondylar notch width,femoral condyle width,intercondylar notch height,Blumensaat's line inclination angle,and lateral tibial posterior slope(P>0.05).(2)Binary logistic regression found that the intercondylar notch width index was associated with posterior cruciate ligament tibial avulsion fracture.(3)It is indicated that patients with posterior cruciate ligament tibial avulsion fracture had smaller intercondylar notch width index,intercondylar notch shape index,intercondylar notch angle,and medial tibial posterior slope compared with those with a normal posterior cruciate ligament.The intercondylar notch width index was an independent risk factor for tibial avulsion fracture of the posterior cruciate ligament.

Abstract:BACKGROUND:Mesenchymal stem cells have shown good therapeutic effects in ischemic stroke,while the role of multilineage-differentiating stress-enduring(Muse)cells isolated from adipose-derived mesenchymal stem cells in ischemic stroke needs further study. OBJECTIVE:To explore the neurorestorative effect of intravenous administration of adipose Muse cells on ischemic stroke in rats. METHODS:The Muse cells expressing stage-specific embryonic antigen 3 were sorted by magnetic beads after long-term(4 hours)trypsin incubation of human adipose-derived mesenchymal stem cells.The middle cerebral artery occlusion model was established in rats.After successful modeling,the adipose-derived mesenchymal stem cell group and adipose-derived Muse group were injected with 200 μL adipose-derived mesenchymal stem cell suspension or adipose-derived Muse cell suspension(containing 2×105 cells)via the tail vein.The saline group was injected with 200 μL saline.Behavioral scores of rats were evaluated at 3 and 7 days after injection.At 3 days after injection,hematoxylin-eosin staining was used to detect brain tissue damage.Immunofluorescence was used to detect the expression of microtubule-associated protein 2 and Ki67 in the brain tissue injury area.TUNEL staining was used to observe cell apoptosis in the injury area.Western blot assay was used to detect the protein expression of growth-associated protein 43,Bcl2,and Bax. RESULTS AND CONCLUSION:(1)After magnetic bead sorting,the flow cytometry showed that the expression rate of stage-specific embryonic antigen 3 was as high as 80%.(2)At 7 days after injection,the neurological function injury scores of the adipose mesenchymal stem cell group and the adipose Muse cell group were reduced compared with those of the saline group(P<0.05).At 3 days after cell transplantation,the neurological function injury scores of the adipose Muse cell group were reduced compared with those of the adipose mesenchymal stem cell group(P<0.05).(3)Hematoxylin-eosin staining showed that the inflammatory response and vacuolation in the cerebral cortex of rats in the adipose mesenchymal stem cell group and the adipose Muse cell group were alleviated.Microtubule-associated protein 2 fluorescence staining showed that the adipose Muse cell group inhibited neuronal loss more significantly.(4)TUNEL staining and Ki67 fluorescence staining results showed that apoptosis of cells in the injured area of the adipose mesenchymal stem cell group and the adipose Muse cell group was reduced,and cell proliferation was increased.(5)Western blot assay results showed that the Bcl-2/Bax ratio and growth-associated protein 43 protein expression in the adipose mesenchymal stem cell group and the adipose Muse cell group were increased,and the adipose Muse cell group was superior to the adipose mesenchymal stem cell group in inhibiting cell apoptosis.The results showed that the nerves of rats transplanted with adipose mesenchymal stem cells and adipose Muse cells were well repaired,and Muse cells played a better role in inhibiting cell apoptosis.

Abstract:BACKGROUND:Endplate defects are one of the important causative factors of lumbar degeneration,and their morphological characteristics may significantly affect the local mechanical environment of the spine.However,the effects of their different morphologic defects on the biomechanical properties of the lumbar spine have not been fully elucidated. OBJECTIVE:To investigate the effects of focal marginal defects,focal central defects,and angular defects on the stress distribution of lumbar endplates,intervertebral discs,and small joints,and to reveal their underlying biomechanical mechanisms. METHODS:Lumbar CT images were obtained from a healthy 36-year-old male volunteer,and a complete endplate model of the L4-L5 segment was reconstructed.Three typical endplate defect models(focal marginal defect,focal central defect,and angular defect)were constructed based on the classification of vertebral endplate defects in clinical imaging studies.By applying dynamic loads and corresponding moments to simulate physiological spinal muscle loads and typical motion loads,such as stance,forward flexion,backward extension,lateral bending,and rotation,the biomechanical stress distribution characteristics and peak changes in the vertebral cartilage endplates,intervertebral disc annulus fibrosus,nucleus pulposus,and facet joints during physiological spinal motion were evaluated under different defect types.The mechanisms by which different defect types affect lumbar spine biomechanical stability were explored. RESULTS AND CONCLUSION:(1)Different defect types significantly altered the stress conduction pathways in the endplates and adjacent structures.Edge defects primarily affected the stress distribution on the lateral side of the annulus fibrosus,while central defects significantly altered load bearing capacity during extension.(2)Significant stress gradient concentrations were observed at the defect edges,indicating potential microinjury risk.(3)Angular endplate defects produced significant stress concentrations under dynamic loading,potentially contributing to segmental instability and accelerated degeneration.(4)These results provide a biomechanical basis for elucidating the involvement of endplate defects in intervertebral disc degeneration and facet joint injury,providing valuable guidance for early clinical identification of high-risk defect types and the development of targeted preventive strategies.

Abstract:BACKGROUND:Although cell transplantation offers a promising approach for the treatment of myocardial infarction,the low transplantation rate limits its application.Therefore,promoting the proliferation of transplanted cells and reducing apoptosis are the key issues to be solved urgently to improve the therapeutic effect. OBJECTIVE:To investigate the effects of non-coding RNA-activated by DNA damage(NORAD)on the proliferation of human induced pluripotent stem cell-derived cardiomyocytes and their apoptosis induced by oxygen-glucose deprivation/reoxygenation,as well as the effects of transplanting NORAD-overexpressing human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-NORADOECMs)on cardiac function in a murine model of myocardial infarction. METHODS:The expression of NORAD in the hearts of mice at different ages(3 days old and 8 weeks old)was measured by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).A cellular model of hiPSC-NORADOECMs was established by infecting human induced pluripotent stem cells with a lentiviral vector designed to specifically upregulate NORAD,followed by directed differentiation.These cells were divided into a control group(infected with a control lentivirus)and an experimental group(infected with a NORAD-overexpressing lentivirus).The efficiency of NORAD overexpression was detected by RT-qPCR.The expression level of proliferation marker protein Ki67 was detected by cellular immunofluorescence.Intracellular reactive oxygen species levels were measured by flow cytometry after cell apoptosis was induced by oxygen-glucose deprivation/reoxygenation induction.The expression levels of Bcl-2-associated X protein(Bax),B-cell lymphoma 2(Bcl-2),and cleaved caspase-3 were analyzed by western blot assay.hiPSC-NORADOECMs were transplanted into the infarcted area of the myocardial infarction mouse model.The cardiac function was evaluated by echocardiography four weeks later. RESULTS AND CONCLUSION:(1)NORAD expression was significantly higher in the hearts of 3-day-old neonatal mice compared with 8-week-old adult mice.(2)The differentiation of human induced pluripotent stem cells into spontaneously contracting human induced pluripotent stem cell-derived cardiomyocytes was successfully achieved.(3)A stable human induced pluripotent stem cell-derived cardiomyocytes model overexpressing NORAD(hiPSC-NORADOECMs)was successfully established.(4)The expression of the proliferation marker Ki67 was significantly increased in the NORAD-overexpressing group compared with controls.(5)Compared with controls,NORAD overexpression reduced reactive oxygen species accumulation induced by oxygen-glucose deprivation/reoxygenation in human induced pluripotent stem cell-derived cardiomyocytes;Bax and cleaved caspase-3 levels were decreased,while Bcl-2 expression was upregulated.(6)Transplantation of hiPSC-NORADOECMs significantly enhanced cardiac function in a murine model of myocardial infarction.Collectively,these findings suggest that overexpression of the NORAD promotes the proliferation of human induced pluripotent stem cell-derived cardiomyocytes,reduces reactive oxygen species accumulation,and inhibits apoptosis following oxygen-glucose deprivation/reoxygenation induction,thereby enhancing the cardiac repair capacity of human induced pluripotent stem cell-derived cardiomyocytes in infarcted hearts.

Abstract:BACKGROUND:There are individual differences in the effectiveness of arthroscopic surgery in improving knee osteoarthritis,and subjective scoring scales may be biased in evaluating the clinical efficacy of arthroscopic treatment for knee osteoarthritis. OBJECTIVE:To explore the correlation between gait parameters and joint function recovery in patients with knee osteoarthritis after arthroscopic surgery. METHODS:A total of 98 patients with knee osteoarthritis admitted to Chongqing Armed Police Corps Hospital from October 2023 to October 2024 were selected as the research subjects.According to Lysholm knee function score after 6 months of follow-up,they were divided into the excellent group(n=63)and the fair group(n=35).Clinical data including gender,age,body mass index,disease duration,respiration,heart rate,Kellgren-Lawrence grade,smoking history,drinking history,hypertension history,location of onset,and postoperative complications were collected.The intraoperative and postoperative indicators,as well as the knee joint function scores and gait parameters at different times before and after surgery were compared between the two groups.Multivariate Logistic regression was used to analyze the independent influencing factors of knee joint function recovery.Stratified regression analysis was conducted to explore the impact of different clinical and pathological characteristics after treatment on gait parameters.Generalized estimating equations were used to analyze the differences in gait parameters among patients with different knee joint functions after treatment.Generalized additive models were used to analyze the impact of gait parameters after treatment on the Lysholm score.Receiver operating characteristic curves were drawn to analyze the value of gait parameters in predicting the recovery of knee joint function after treatment. RESULTS AND CONCLUSION:(1)There were statistically significant differences in age,disease duration,Kellgren-Lawrence grade,and postoperative complications between the excellent group and the fair group(P<0.05).(2)The fair group had longer operation time,more intraoperative blood loss,longer time for swelling to subside,and longer rehabilitation time than the excellent group(P<0.05).(3)The fair group had higher Western Ontario and McMaster Universities Osteoarthritis Index and Visual Analogue Scale scores,and lower Lysholm scores,step frequency,and step speed than the excellent group(P<0.05).(4)Logistic regression analysis showed that age,Kellgren-Lawrence grade,postoperative complications,and time for swelling to subside were independent risk factors for knee joint function recovery(P<0.05).(5)Stratified regression analysis showed that age,Kellgren-Lawrence grade,time for swelling to subside,and postoperative complications had negative effects on step frequency and step speed(β<0,P<0.05).(6)Generalized estimating equation analysis showed that there was a correlation between the degree of knee joint function recovery and gait characteristics(β>0,P<0.05).(7)Generalized additive model analysis showed that the impact of step frequency and step speed on the Lysholm score after treatment was linear.(8)Receiver operating characteristic curve analysis showed that the combined detection of step frequency and step speed had higher predictive efficacy(Area under the curve>0.85,P<0.05).(9)It suggests that arthroscopic surgery for knee osteoarthritis can improve knee joint function.By dynamically tracking changes in postoperative gait parameters and combining with functional scoring scales,the correlation between step frequency,step speed,and the outcome of knee joint function can be revealed,further clarifying its clinical predictive value.This provides a new quantitative tool for clinical functional assessment and enables more precise postoperative rehabilitation guidance.

Abstract:BACKGROUND:In recent years,the application of kidney organoid technology in acute kidney injury has gradually become a research hotspot.Traditional animal models have species differences from humans,and physiological and pathological processes of their kidneys cannot fully represent the human situation.Kidney organoid technology forms 3D kidney models through stem cell culture,which can simulate the complex structure and function of human kidneys.It has shown great potential in disease modeling and mechanism exploration of acute kidney injury,prediction of drug nephrotoxicity,and exploration of regeneration and repair mechanisms. OBJECTIVE:To summarize the progress in the application of organoid technology in acute kidney injury and to provide new technical approaches and research strategies for the prevention and treatment of acute kidney injury. METHODS:The relevant literature on organoids and acute kidney injury was retrieved through the China National Knowledge Infrastructure and PubMed databases,in which Chinese and English search terms mainly included"acute kidney injury,organoid,pluripotent stem cell,3D bioprinting,kidney-on-a-chip,regenerative medicine,kidney transplantation."All retrieved literature included original research articles and relevant reviews.The search period covered the establishment of each database to April 2025.A total of 99 articles were finally selected for analysis and summary. RESULTS AND CONCLUSION:(1)The cell sources reported in the current literature for inducing the formation of kidney organoids mainly include pluripotent stem cells,embryonic stem cells,and stem cells derived from urine.These kidney organoids play a significant role in in vitro drug screening,kidney development,and disease modeling.(2)3D bioprinting and chip kidney technology are emerging techniques for constructing kidney-like organs.3D bioprinting can precisely and specifically construct complex multi-cellular structures.Chip kidney technology has features such as high gas permeability,sensitivity,and low cost,which can extend the lifespan of the organoids and increase biocompatibility.It is suitable for preclinical drug development and toxicity screening.(3)The combination of gene editing technology and kidney organoid models has brought new perspectives and tools for the study of kidney diseases,drug development,and regenerative medicine.This technology can construct kidney organoids with specific reporter genes or sensitive indicators,and can also amplify and classify specific types of kidney cells in the organoids.(4)Renal organoids demonstrate unique advantages in disease simulation,drug evaluation,and exploration of regenerative treatment strategies.They can serve as in vitro models for the toxic mechanisms of drugs such as cisplatin,doxorubicin,and red yeast rice supplements that cause acute kidney injury.They can also be used to screen high-throughput drugs and therapeutic drug targets,playing a significant role in the field of regenerative medicine for kidney transplantation.

Abstract:BACKGROUND:In patients with severe medial compartment osteoarthritis of the knee combined with moderate lateral compartment osteoarthritis,it is controversial whether medial monondylar replacement alone can achieve satisfactory clinical benefit. OBJECTIVE:To compare and analyze the difference in clinical outcomes of unicompartmental knee arthroplasty and total knee arthroplasty in patients with heterogeneous degenerative changes of the knee(medial compartment Kellgren-Lawrence grade III-IV combined with lateral compartment Kellgren-Lawrence grade II). METHODS:Knee arthroplasty patients with severe medial ventricular osteoarthritis combined with mild and moderate lateral ventricular osteoarthritis shown by preoperative knee X-ray were selected and divided into unicompartmental knee arthroplasty group and total knee arthroplasty group according to different operation methods,and 50 patients were included in each group according to 1:1 pairing.Western Ontario and McMaster University Osteoarthritis Index(WOMAC),American Knee Association Score,joint amnesia score,postoperative gait parameters,and complications were collected at 3,6 months,1 and 2 years after surgery,and the relevant data were statistically analyzed to compare the differences in postoperative efficacy between the two groups. RESULTS AND CONCLUSION:(1)The American Knee Association Score and WOMAC scores of the unicompartmental knee arthroplasty group were better than those of the total knee arthroplasty group at 3,6 months and 1 year after surgery(P<0.05).Two years after surgery,American Knee Association Score and WOMAC score were not significantly different between two groups(P>0.05).(2)There was no significant difference in joint amnesia score between the two groups 3 months after operation(P>0.05).The joint amnesia score was higher in the unicompartmental knee arthroplasty group than that in the total knee arthroplasty group at 6 months,1 year and 2 years after surgery(P<0.05).(3)At 1 year after surgery,step speed and step length were better in the unicompartmental knee arthroplasty group than those in the total knee arthroplasty group(P<0.05).No statistically significant differences in step frequency were detected between the two groups(P>0.05).(4)No complications occurred in both groups within 2 years after operation.(5)Follow-up showed that unicompartmental knee arthroplasty was clinically equivalent to total knee arthroplasty in patients with severe medial compartment osteoarthritis(Kellgren-Lawrence III-IV)combined with moderate lateral compartment degeneration(Kellgren-Lawrence II)2 years after surgery.Unicompartmental knee arthroplasty had a significant biomechanical advantage within 1 year after surgery.No significant progression of lateral interventricular osteoarthritis was observed during follow-up(Kellgren-Lawrence grade increase),but further long-term observation is needed.Before making decisions,doctors should establish a multi-dimensional evaluation framework(including patient age,exercise load expectation,unicompartmental knee arthroplasty operation amount of the surgeon,etc.),and fully inform patients of the risk of lateral compartment progression,and decide whether to perform unicompartmental knee arthroplasty after comprehensive assessment.

Abstract:BACKGROUND:Periodontitis is a highly prevalent chronic inflammatory disease.Previous research has predominantly focused on specific immune cells or cytokines.Consequently,systematically elucidating its immune mechanisms and discovering novel therapeutic targets hold significant implications. OBJECTIVE:To analyze the expression profiles of periodontitis-associated immune cell subpopulations and identify key differentially expressed genes with a causal relationship to the disease,thereby exploring potential molecular mechanisms and key genes involved in periodontitis and immune cell dynamics. METHODS:Single-cell RNA sequencing data from the GEO database were used to analyze immune cell subset heterogeneity and identify differentially expressed genes.Mendelian randomization analysis was performed using expression quantitative trait loci data to infer causal relationships between immune cell gene expression and periodontitis risk.Pathway enrichment and immune infiltration analyses were performed on the identified causal genes to reveal the associations between differentially expressed genes and immune cells with the development and progression of periodontitis.CellChat trajectory analysis was used to explore intercellular communication.To validate key findings,gingival tissue samples were collected from 20 patients with periodontitis diagnosed by the Department of Stomatology at The First Affiliated Hospital of Shihezi University(periodontitis group)and 20 healthy gingival tissue samples from patients undergoing orthodontic or impacted tooth extraction(control group).RT-qPCR and immunohistochemistry were used to examine the expression of key genes. RESULTS AND CONCLUSION:Comprehensive analysis identified 23 immune cell clusters in periodontitis and three key genes(annexin A1,solute carrier family 11 member 1,and vimentin)that were significantly causally associated with periodontitis risk.Pathway enrichment analysis revealed their involvement in key immune regulatory mechanisms.Further analysis and characterization of immune subtype receptor ligands and key cell subtype trajectories revealed distinct roles for annexin A1,solute carrier family 11 member 1,and vimentin in disease progression.Annexin A1,solute carrier family 11 member 1,and vimentin mRNA expression levels were upregulated in periodontitis tissues compared with healthy controls(P<0.05).This study reveals the key role of immune cell subsets in periodontitis and validates key genes(annexin A1,solute carrier family 11 member 1,and vimentin)with a causal relationship with periodontitis.

Abstract:BACKGROUND:Stem cells show a great potential in oral tissue regeneration but face challenges such as immune rejection and tumor formation.Exosomes are nanoscale extracellular vesicles secreted by cells,reducing immunogenicity and tumor risks while maintaining stem cell functions,such as promoting angiogenesis and tissue repair. OBJECTIVE:To summarize the mechanisms and roles of exosomes in oral tissue regeneration,explore exosome engineering strategies and the challenges and future directions in the application of exosomes in oral regenerative medicine. METHODS:The relevant literature published from the WanFang and PubMed databases from their inception to 2025 was searched using Chinese search terms"stem cells,exosomes,dental pulp regeneration,periodontal regeneration"and English search terms"exosomes,stem cells,dentistry,regenerate."Finally,94 articles were included for review and analysis. RESULTS AND CONCLUSION:(1)Exosomes have lower immunogenicity and no tumorigenic risk compared with stem cells.They are more stable and easier to store and transport.Additionally,exosomes can penetrate dense tissues for targeted delivery.They can avoid the ethical and immune rejection issues faced by stem cell therapy,making them a safer and more effective treatment option.(2)Exosomes have shown significant efficacy in the regeneration of oral tissues,including dental pulp,periodontal tissue,craniofacial bone,salivary glands,nerves,and skin.They can promote tissue repair and regeneration through multiple mechanisms,demonstrating broad application prospects.(3)Engineering strategies such as pre-treatment,isolation and purification,and targeted modification can enhance the functions of exosomes,improving their therapeutic potential and clinical feasibility.However,the current technologies still have limitations.Further optimization is needed in the future to promote their widespread application.

Abstract:BACKGROUND:A combination of subjective and objective evaluation criteria is often required to more accurately and comprehensively assess knee function in patients undergoing anterior cruciate ligament reconstruction. OBJECTIVE:To review the evolving trends in effectiveness and safety evaluation criteria after anterior cruciate ligament reconstruction and analyze the dynamic shift in the use of subjective and objective assessment tools. METHODS:A systematic search of PubMed and Embase was conducted up to August 22,2023 to identify studies assessing knee function after anterior cruciate ligament reconstruction.A total of 136 eligible studies meeting the inclusion criteria were included.The frequency of each evaluation standard was extracted and analyzed over time using Origin 2025 software. RESULTS AND CONCLUSION:(1)Between 1990 and 2005,objective measures were widely applied.Since 2005,subjective scoring systems,particularly patient-reported outcome measures,have increased sharply,surpassing objective standards in frequency from 2009 onward.(2)Early use was dominated by the Lysholm scale and Tegner activity score,while the International Knee Documentation Committee-Subjective Knee Form,Knee Injury and Osteoarthritis Outcome Score,and anterior cruciate ligament-return to sport after injury gradually emerged as the main tools in later years.(3)In contrast,objective assessments such as the KT1000/2000 arthrometer,Lachman test,and hop tests remained relatively stable but showed an overall downward trend.(4)This suggests that the evaluation criteria for the effectiveness and safety of anterior cruciate ligament reconstruction have shifted from a primary focus on objective knee stability to an emphasis on patient-reported experiences.(5)This study is the first to quantitatively reveal the dynamic evolution of mainstream assessment tools.Current clinical practice highlights the integration of subjective and objective measures,and we recommend the following combination:International Knee Documentation Committee-Subjective Knee Form or Knee Injury and Osteoarthritis Outcome Score,together with anterior cruciate ligament-return to sport after injury,plus either the KT1000/2000 arthrometer or hop tests.Such a framework provides a more comprehensive reflection of both functional recovery and patient perception,offering an evidence-based foundation for the development of future integrated evaluation systems.

Abstract:BACKGROUND:Age-related degeneration is closely associated with bone metabolic imbalance.In the jaw,this manifests as alveolar bone resorption,tooth loosening,and even loss.Impaired osteogenic differentiation potential of senescent jaw bone marrow mesenchymal stem cells is a critical factor hindering jaw bone regeneration.Quercetin,a natural flavonoid compound,exhibits antioxidant,anti-inflammatory,and cell differentiation-regulating properties,yet effect and mechanism of quercetin in osteogenic differentiation of senescent jaw bone marrow mesenchymal stem cells remain unclear. OBJECTIVE:To investigate the effects of quercetin on the proliferation,migration,osteogenic differentiation,and senescence of aged jaw bone marrow mesenchymal stem cells. METHODS:Jaw bone marrow mesenchymal stem cells were isolated from the mandibles of 10 8-week-old SD rats and cultured using a combination of bone marrow flushing and bone slice digestion.Jaw bone marrow mesenchymal stem cells were subcultured to the third and seventh passages,serving as the young and senescent cell groups,respectively.Quercetin was then added to the senescent cell group.The CCK-8 assay was used to assess the effects of 0.01,0.1,1,10,and 100 μmol/L quercetin solutions on the proliferation of senescent jaw bone marrow mesenchymal stem cells to identify the optimal quercetin concentration.Cell migration ability was assessed by cell scratch test.RT-qPCR and western blot assay were used to examine the expression of senescence markers.β-Galactosidase staining was used to assess the proportion of cells expressing these markers.Seven days after osteogenic induction,the expression of osteogenic-related markers was assessed by RT-qPCR and western blot assay.Alkaline phosphatase staining was performed on day 14 of osteogenic induction.Alizarin red staining was performed on day 21 of osteogenic induction.Western blot assay was used to assess the expression of phosphorylated protein kinase B,protein kinase B,phosphorylated mammalian target of rapamycin,and mammalian target of rapamycin. RESULTS AND CONCLUSION:(1)Compared with the young cell group,the proliferation ability of the senescent cell group was decreased.Compared with the senescent cell group,1 μmol/L quercetin significantly promoted the proliferation of senescent jaw bone marrow mesenchymal stem cells(P<0.01).(2)Compared with the senescent cell group,the migration ability of senescent jaw bone marrow mesenchymal stem cells was improved;the proportion of β-galactosidase-positive cells was significantly decreased,and the expression of senescence-related P16,P53,and P21 mRNA and protein was decreased in the quercetin group(P<0.05).(3)After osteogenic induction,compared with the senescent cell group,the calcium nodule formation ability,alkaline phosphatase staining area,and the mRNA and protein expressions of alkaline phosphatase,osteopontin,and Runt-related transcription factor 2 were increased in the quercetin group(P<0.05).(4)Compared with the senescent cell group,the phosphorylation levels of protein kinase B and mammalian target of rapamycin were significantly decreased in the quercetin group(P<0.05).These results suggest that quercetin inhibits multiple-passage senescence of jaw bone marrow mesenchymal stem cells and promotes osteogenic differentiation by regulating the protein kinase B/mammalian target of rapamycin signaling pathway.

Abstract:BACKGROUND:Studies showed that multiple myeloma microenvironment has the function of inducing mesenchymal stem cells to become senescent phenotype,while the effect of senescent bone marrow mesenchymal stem cells on multiple myeloma cells is rarely reported. OBJECTIVE:To investigate the effect of senescent bone marrow mesenchymal stem cells on the proliferation of multiple myeloma cells through paracrine galectin-3. METHODS:Bone marrow blood was collected from healthy donors,and bone marrow mesenchymal stem cells were extracted by Ficoll density gradient centrifugation and adherent purification.The third-generation bone marrow mesenchymal stem cells were taken and induced with 200 µmol/L hydrogen peroxide solution for 2 hours.After the treatment was completed,they were replaced with L-DMEM complete medium for 24 hours of culture to construct the senescent bone marrow mesenchymal stem cell model.β-Galactosylase staining and senescence gene P21 were used for identification.Meanwhile,the expression of galectin-3 in aging bone marrow mesenchymal stem cells was detected by RT-qPCR.The supernatant of senescent bone marrow mesenchymal stem cells was collected and the conditioned medium of senescent bone marrow mesenchymal stem cells was prepared by centrifugal concentration.After culturing the multiple myeloma cell line U266 for 24 hours with it,the proliferation of U266 cells was detected by CCK-8 assay.The apoptosis of U266 cells was detected by flow cytometry.The expression levels of BCL-2 protein and mRNA in U266 cells were detected by RT-qPCR and western blot assay.Bone marrow blood was collected from patients with multiple myeloma and healthy individuals.The level of galectin-3 was detected by ELISA. RESULTS AND CONCLUSION:(1)After hydrogen peroxide induction,the number of cells positive for β-galactosidase staining increased significantly,and P21 and galectin-3 mRNA expression levels were upregulated(P<0.01).(2)Compared with the control group,24 hours of culture of U266 cells with conditioned medium from senescent bone marrow mesenchymal stem cells increased the level of cell proliferation(P<0.05),decreased the apoptosis rate(P<0.05),and rose the expression levels of BCL-2 protein and mRNA(P<0.05).(3)Galectin-3 levels in the bone marrow of patients with multiple myeloma were significantly higher than those in healthy controls(P<0.05).These results suggest that senescent bone marrow mesenchymal stem cells may promote the proliferation of multiple myeloma cells by paracrine galectin-3,which in turn upregulates BCL-2 expression.

Abstract:BACKGROUND:Breast cancer organoids,as a novel in vitro model,can not only simulate the biological characteristics of breast cancer but also to some extent reproduce the impact of the tumor microenvironment on the tumor,facilitating research on breast cancer and further promoting precision medicine. OBJECTIVE:To review the application status of breast cancer organoids in the field of therapeutic drugs,including chemotherapy,targeted therapy,and immunotherapy,over the past few years,and to discuss the existing limitations in order to further promote their application in breast cancer treatment. METHODS:The first author conducted a search in the China National Knowledge Infrastructure(CNKI)and PubMed databases in June 2025,with the publication time of the literature ranging from January 2010 to June 2025.The Chinese and English search terms included"organoid,breast organoid,breast cancer organoid,breast cancer experimental model,precision medicine,targeted therapy,chemotherapy,immunotherapy,drug sensitivity."A total of 58 articles were included in the review. RESULTS AND CONCLUSION:(1)Compared with traditional breast cancer cell experiments that lack validation of tissue structure,cell-to-cell interactions,and in vivo microenvironments,breast cancer organoids,with their diverse origins from primary tumor cells and continuous innovation in culture systems,can simulate cell-to-cell interactions and reproduce the biological characteristics of breast cancer and its tumor microenvironment.This makes breast cancer organoids one of the most promising tools in the field of breast cancer research and provides greater potential for improving treatment resistance in clinical breast cancer patients through drug sensitivity screening.(2)The application of drug sensitivity trial results from breast cancer organoids in clinical practice has yielded promising outcomes.By conducting drug sensitivity tests with common chemotherapeutic agents,targeted drugs,and immunotherapeutic agents in breast cancer organoids,the mechanisms of drug action against tumors and the synergistic effects of multiple drugs are validated.This approach helps avoid the use of primarily resistant and highly toxic treatment drugs in patients,enabling the formulation of personalized treatment plans and evidence-based precision medical strategies.(3)Breast cancer organoids still face limitations in practical applications,such as low success rates in model construction,difficulties in model growth,and the lack of angiogenesis processes.To overcome these limitations,it is necessary to increase the amount of source tissue,improve culture systems,and innovate culture techniques.These improvements will facilitate comprehensive therapeutic drug selection through breast cancer organoids and assist in the development of personalized precision medicine.

Abstract:BACKGROUND:Transforaminal lumbar interbody fusion is one of the main surgical methods for treating degenerative lumbar diseases such as lumbar disc herniation,lumbar spinal stenosis,and lumbar spondylolisthesis.Hidden blood loss refers to the concealed loss of blood volume in patients,which is often overlooked by people. OBJECTIVE:To evaluate the perioperative blood loss during transforaminal lumbar interbody fusion using a new method,calculate the hidden blood loss based on the new method,and analyze its risk factors. METHODS:The medical records of 93 patients with lumbar degenerative diseases(lumbar spinal stenosis,lumbar disc herniation,and lumbar spondylolisthesis)who were hospitalized in the Department of Spine Surgery of Second Affiliated Hospital of Soochow University from October 2023 to October 2024 were retrospectively analyzed.The general data of patients were collected,such as age,gender,height,body mass,body mass index,and whether they had hypertension and diabetes;surgical data,such as the number of surgical segments,operation time,and American Society of Anesthesiologists anesthesia grade;laboratory tests,such as prothrombin time,activated partial thromboplastin time,international normalized ratio,platelet count,fibrinogen,and D-dimer level.Pearson or Spearman correlation analysis was used to explore the correlation between patient characteristics and postoperative hidden blood loss,and multivariate linear regression analysis was utilized to determine the independent risk factors for postoperative hidden blood loss. RESULTS AND CONCLUSION:(1)The average hidden blood loss calculated by the new method for transforaminal lumbar interbody fusion was(284.24±352.76)mL,accounting for 58.6%of the total blood loss;while the average hidden blood loss calculated by the traditional surgical method was(165.77±339.89)mL,accounting for 34.15%of the total blood loss.The difference between the two was significant(P<0.05).(2)In the univariate analysis,hidden blood loss was significantly correlated with the number of segments(r=0.213,P=0.040)and operation time(r=0.210,P=0.043)and negatively correlated with platelet count(r=-0.324,P=0.018).(3)In the multivariate linear regression analysis,decreased platelet count was an independent risk factor for hidden blood loss(P=0.016).(4)It is concluded that the new method was more accurate in estimating hidden blood loss,and hidden blood loss was an important part of the total blood loss during the perioperative period.Increased number of segments,prolonged operation time,and decreased platelet count were risk factors for hidden blood loss in transforaminal lumbar interbody fusion,among which decreased platelet count was an independent risk factor.

Abstract:OBJECTIVE:To conduct a meta-analysis concerning the effects of human umbilical cord blood mesenchymal stem cells on pain and function in patients with knee osteoarthritis. METHODS:Using the Chinese search terms"human umbilical cord blood,mesenchymal stem cells,knee joint-related diseases"and the English search terms"human cord blood,mesenchymal stem cell,MSC,knee osteoarthritis,knee joint disease,knee joint disorders,knee OA,"we conducted searches in the CNKI,WanFang,VIP,PubMed,Elsevier,and Web of Science databases.The search timeframe spanned from the establishment of each database until June 13,2024.The quality of the included literature was assessed using the Cochrane Risk of Bias tool and the ROBINS-I tool.For meta-analysis,the Revman software was utilized,calculating mean differences for continuous variables and relative risks for dichotomous variables,along with 95%confidence intervals. RESULTS:Three randomized controlled trials and three case-control studies involving a total of 248 subjects were included,with the quality of the literature evaluated as moderate.The results of the meta-analysis showed that(1)visual analog scale score was significantly lower in the trial group than in the control group,with significant differences(χ2=44.98,P<0.001,I²=91%).(2)The Western Ontario and McMaster Universities Osteoarthritis Index score was significantly lower in the trial group than in the control group,with significant differences(χ2=16.84,P<0.001,I²=88%).(3)The Lysholm scoring system score was significantly higher in the trial group than in the control group,with significant differences(χ2=0.12,P=0.73,I²=0%).(4)The incidence of adverse reactions was significantly higher in the trial group than in the control group,with significant differences(χ2=4.99,P<0.001,I²=20%),with a pooled risk difference of 0.21,corresponding to a number needed to treat of 5. CONCLUSION:Human umbilical cord blood mesenchymal stem cells can relieve pain in patients with knee osteoarthritis,improve knee function,and achieve a good balance between safety and effectiveness.

Abstract:BACKGROUND:Rutin can effectively prevent osteoporosis,but its mechanism of action remains unclear. OBJECTIVE:To investigate the effect of rutin on osteogenesis of MC3T3-E1 cells under the action of neutrophil extracellular traps. METHODS:(1)Human myeloid leukemia dHL60 cells were stimulated with phorbol 12-myristate 13-acetate to induce neutrophil extracellular trap formation in vitro.dHL60 cells were divided into four groups and cultured.The control group received Hank's balanced salt solution,while the other three groups received 50 nmol/L phorbol esters.The latter two groups were then treated with either 250 μmol/L rutin or 250 μmol/L rutin and 5 U/mL DNase I.dHL60 cell apoptosis was assessed by flow cytometry,and the expression of marker genes and proteins associated with neutrophil extracellular trap formation was determined by RT-qPCR and western blot assay.(2)MC3T3-E1 cells were divided into six groups for culture:the control group was treated with Hank's balanced salt solution,and the other five groups were treated with 50 nmol/L phorbol esters;dHL60 cells and 50 nmol/L phorbol esters;100 μmol/L rutin;dHL60 cells,50 nmol/L phorbol esters,and 250 μmol/L rutin;and dHL60 cells,50 nmol/L phorbol esters,250 μmol/L rutin,and 5 U/mL DNase I.Flow cytometry was used to assess apoptosis in MC3T3-E1 cells treated with neutrophil extracellular traps.Alkaline phosphatase staining and Alizarin red staining were used to determine the osteogenic and mineralization abilities of MC3T3-E1 cells treated with neutrophil extracellular traps.RT-qPCR and western blot assay were used to examine the expression of osteogenesis-related genes and proteins in MC3T3-E1 cells treated with neutrophil extracellular traps. RESULTS AND CONCLUSION:(1)Compared with the blank control group,rutin significantly inhibited the mRNA and protein expressions of protein arginine deiminase 4,myeloperoxidase,and neutrophil elastase in dHL60 cells(P<0.000 1).Compared with the rutin group alone,the combined intervention of rutin and DNase I had a more significant downregulation effect on protein arginine deiminase 4,myeloperoxidase,and neutrophil elastase(P<0.05),indicating that rutin could significantly inhibit the formation of neutrophil extracellular traps.(2)After dHL60 cells induced neutrophil extracellular traps and then co-cultured with MC3T3-E1 cells,the mRNA and protein expressions of Runt-related transcription factor 2,β-catenin,and bone morphogenetic protein 2 in MC3T3-E1 cells were significantly downregulated(P<0.000 1),and the apoptosis rate was significantly increased(P<0.000 1),which indicate that neutrophil extracellular traps significantly inhibit the osteogenic capacity of MC3T3-E1 cells in vitro and promote their apoptosis.Rutin alone or combined with DNase I significantly improved the apoptosis and osteogenic capacity of MC3T3-E1 cells exposed to neutrophil extracellular traps,and the combined effect of rutin and DNase I was more pronounced than that of rutin alone,suggesting that rutin may inhibit the formation of neutrophil extracellular traps,thereby improving the osteogenic capacity of MC3T3-E1 cells.(3)Molecular docking and molecular dynamics simulations revealed that rutin binds well to the target proteins of protein arginine deiminase 4,myeloperoxidase,and neutrophil elastase,indicating that rutin can specifically inhibit the formation of neutrophil extracellular traps.

Abstract:BACKGROUND:Currently,the biomechanical differences between cervical rotation manipulation and cervical rotation-traction manipulation for the treatment of cervical radiculopathy have not been systematically elucidated. OBJECTIVE:To compare the biomechanical differences between cervical rotation manipulation and cervical rotation-traction manipulation in the treatment of cervical spondylotic radiculopathy caused by cervical disc herniation,and to provide a basis for the rational selection of manipulation in clinical practice. METHODS:A 27-year-old Asian male patient with cervical spondylotic radiculopathy caused by left posterior cervical disc herniation compressing the nerve root was recruited.The CT scan data of the skull and cervical spine were extracted to construct a finite element model of the skull and the whole cervical spine.After the model was verified,the key parameters of cervical rotation manipulation and cervical rotation-traction manipulation were loaded into the model,and the effects of these two manipulations on the stress of the intervertebral disc,facet joint,spinal cord and nerve root,disc displacement,as well as the volume of the cervical intervertebral foramen and other indexes were compared and analyzed. RESULTS AND CONCLUSION:(1)In terms of Von-Mise stress,the maximum stresses of cervical rotation manipulation on the annulus fibrosus of the intervertebral disc,nucleus pulposus and facet joint were 0.903,0.139,and 2.186 MPa,respectively,which were increased by 18%,13%,and 30%compared with those of cervical rotation-traction manipulation(0.765,0.123,and 1.682 MPa).The maximum stress of the spinal cord and nerve root was 2.547 Mpa,which was 7%lower than that of cervical rotation-traction manipulation(2.738 MPa).(2)In terms of displacement,the maximum forward displacement of the side with intervertebral disc herniation under cervical rotation manipulation was 1.067 mm,which was 11.1%more than that under cervical rotation-traction manipulation(0.960 mm).(3)In terms of the change in the volume of the intervertebral foramen,both manipulations led to an increase compared with before the manipulation.The volume increased by 15.5%under rotation manipulation and 19.8%under rotation-traction manipulation,and the latter had a more significant effect on expanding the volume of the intervertebral foramen.(4)It is concluded that cervical rotation manipulation has an advantage in promoting the forward displacement of the herniated intervertebral disc,but it generates higher stress on the intervertebral disc and facet joint,and is likely to cause intervertebral disc injury.Cervical rotation-traction manipulation makes the spinal cord and nerve root bear slightly higher stress,but it can more effectively expand the volume of the intervertebral foramen and reduce the risk of intervertebral disc structural injury.In clinical treatment,the advantages and disadvantages of these two manipulations should be carefully weighed according to the specific condition of the patient,and the application should be rationally selected.

Abstract:BACKGROUND:In recent years,microRNA(miRNA)has received extensive attention in the pathogenesis and diagnosis and treatment of cerebral small vessel disease,and is involved in the regulation of various pathological processes of cerebral small vessel disease. OBJECTIVE:To review the role of miRNA in the pathogenesis,diagnosis,and treatment of cerebral small vessel disease,and to provide effective therapeutic targets and new potential biomarkers for the early diagnosis of cerebral small vessel disease. METHODS:"microRNA,cerebral small vessel disease,blood-brain barrier,chronic cerebral hypoperfusion,inflammation,apoptosis,diagnosis,biomarkers"were used as English search terms for PubMed search."Exosomal miRNA,cerebral small vessel disease"were used as Chinese search terms for CNKI search.The search time limit was from inception to January 2025.Through the preliminary screening of reading titles and abstracts,the literature with poor relevance and duplicate content was excluded,and finally 72 articles were included for inductive discussion. RESULTS AND CONCLUSION:(1)Through the excavation and discussion of the biological functions and characteristics of exosomal miRNAs,it was confirmed that exosomal miRNAs are important related components in the occurrence and progression of cerebral small vessel disease diseases.(2)Exosomal miRNA participates in the regulation of various pathological processes of cerebral small vessel disease,and plays an important role in the pathological mechanism of cerebral small vessel disease by protecting the blood-brain barrier,improving chronic cerebral hypoperfusion,reducing inflammatory response,and inhibiting apoptosis.(3)Exosomal miRNA can effectively target different pathological links of cerebral small vessel disease by targeting multiple signaling pathways to intervene at different stages of pathological development.(4)The combination of multiple exosomal miRNAs can effectively improve the disease process of cerebral small vessel disease,and the regulatory effects of different miRNAs play different roles in each stage of the pathological mechanism,and the construction of miRNA action network is of great significance for regulating the development of cerebral small vessel disease.(5)Exosomal miRNAs are widely and stably present in various body fluids,and their specific and significant expression in urine,serum,blood and other body fluids of cerebral small vessel disease patients can be used as an effective basis for the diagnosis of cerebral small vessel disease patients.(6)At present,the main clinical treatment method is to inject miRNA mimics or antagonists to regulate the expression of downstream target genes,and how to exert the most effective therapeutic effect of miRNA still needs to be further studied.(7)As exosome contents,miRNA has great application prospects in the treatment of cerebral small vessel disease.In the future,its mechanism of action should be further explored to provide effective ideas and further optimize the clinical treatment of cerebral small vessel disease.

Abstract:BACKGROUND:Platelets,crucial blood components,are commonly discarded along with white blood cells in traditional blood processing and become medical waste.Optimizing whole blood separation processes to prepare platelet lysate products and exploring their applications in tissue engineering and regenerative medicine are of great value. OBJECTIVE:To optimize blood separation for therapeutic-grade platelet lysate production and explore the protective impact of platelet lysate on myocardial hypoxia injury. METHODS:Twenty-one platelets were isolated from whole blood using a closed blood bag and tubing system,followed by the preparation of twenty-one platelet lysates through the freeze-thaw method.The concentrations of platelet-derived growth factors(platelet-derived growth factor AA,platelet-derived growth factor BB,and platelet-derived growth factor AB),vascular endothelial growth factor,epidermal growth factor,insulin-like growth factor,fibroblast growth factor,and transforming growth factor-β1 in the platelet lysate were quantified using an enzyme-linked immunosorbent assay kit.Detection ranges for these factors were determined.Bacterial and mycoplasma contamination were observed using the colony culture method and a Mycoplasma PCR detection kit,respectively.A hypoxic cardiomyocyte model was established to evaluate the protective effects of platelet lysate on hypoxia-stressed cardiomyocytes. RESULTS AND CONCLUSION:(1)The concentration ranges of major growth factors and cytokines in platelet lysates were as follows:platelet-derived growth factor AA:12.86-24.17 μg/L,platelet-derived growth factor BB:0.25-0.32 μg/L,platelet-derived growth factor AB:85.09-114.91 μg/L,vascular endothelial growth factor:10.57-58.37 μg/L,epidermal growth factor:0.43-0.69 μg/L,insulin-like growth factor 1:106-204.9 μg/L,fibroblast growth factor:0.03-0.06 μg/L,transforming growth factor β1:124.17-192.38 μg/L.(2)Colony culture and mycoplasma detection yielded negative results.(3)The efficiency of cardiomyocyte proliferation was highest with a low concentration(1%)of platelet lysate in culture.A 1%concentration of platelet lysate effectively stimulated cardiomyocytes to produce high levels of superoxide dismutase and glutathione peroxidase,providing protective effects for cardiomyocytes.This study established a method for preparing therapeutic-grade platelet lysate by optimizing the whole blood separation process,thereby improving the utilization of blood resources.Platelet lysate,rich in essential growth factors,significantly promotes the repair of hypoxic injuries in cardiomyocytes.

Abstract:BACKGROUND:The principal compressive and tensile trabecular bones in the proximal femur are essential structures responsible for transmitting forces through the hip joint.Given their complex and heterogeneous composition,exploring their microstructural variations will help improve our understanding of principal stress trabeculae. OBJECTIVE:To extract trabecular microstructural information from various regions of femoral head specimens and study their morphometric patterns,elucidate the microstructural variations of principal stress trabeculae,and provide a theoretical basis for the design of future bionic hip prostheses. METHODS:Totally 11 fresh femoral heads were obtained from patients with femoral neck fractures.They were scanned by Micro-CT and reconstructed by image analysis software(InveonTM Acquisition Workplace).The principal compressive and tensile trabecular bones were segmented into five regions respectively.After selecting the volumes of interest for each region,morphometric analyses were subsequently performed on bone volume fraction,bone surface area fraction,trabecular thickness,trabecular number,trabecular spacing,trabecular pattern factor,and grayscale value. RESULTS AND CONCLUSION:(1)Among the principal compressive trabeculae,Region 5 exhibited the highest bone volume fraction,trabecular thickness,and grayscale value,followed by Region 4 and 1,with statistically significant differences between Region 5 and 1(all P<0.05).Moving from Region 3 to Region 2,1,and Regions 4,5,there was a decreasing trend in bone surface area fraction and trabecular number,accompanied by an increasing trend in trabecular thickness.(2)Among the principal tensile trabeculae,Region 3 displayed the highest values for bone volume fraction,trabecular thickness,trabecular number,and grayscale value,while Region 2 showed opposite trends,with statistically significant differences between the two regions,(excluding grayscale value;all P<0.001).(3)This study proposes a reliable method for extracting the volume of interest of principal stress trabeculae and introduces the novel concept that principal compressive trabeculae represent a functionally graded porous structure.These findings contribute new insights into the anatomical microstructure of principal compressive trabeculae and are expected to guide the future design of bionic hip prostheses.