BACKGROUND:Sarcopenia is an age-related condition characterized by the loss of skeletal muscle mass,strength,and/or physical function.Currently,effective treatments for sarcopenia remain limited.A new therapeutic approach to improve symptoms and prognosis of sarcopenia patients clinically was important.OBJECTIVE:To explore the effects of canine adipose-derived mesenchymal stem cells and their exosomes on a dexamethasone-induced sarcopenia in mice.METHODS:Mesenchymal stem cells were isolated and cultured from canine adipose tissue,and identified and functionally evaluated through flow cytometry and differentiation assays for osteogenesis,adipogenesis,and chondrogenesis.Subsequently,exosomes from adipose-derived mesenchymal stem cells were extracted and characterized using transmission electron microscopy,western blot assay,and nanocoulter tracking analysis.In vitro,the effects of canine adipose-derived mesenchymal stem cells and their exosomes on myotube growth and the expression of muscle atrophy-related genes were investigated using dexamethasone-induced C2C12 myotube atrophy and aging C2C12 models.In vivo,a dexamethasone-induced mouse sarcopenia model was established and received intraperitoneal or intravenous injection of canine adipose-derived mesenchymal stem cells.Therapeutic efficacy was assessed through mouse rotarod performance,histopathological analysis,and muscle atrophy-related genes testing.RESULTS AND CONCLUSION:(1)The isolated canine adipose-derived mesenchymal stem cells highly expressed CD73,CD90,and CD105,and lowly expressed MHC-Ⅱ,CD14,CD19,CD34,and CD45,and successfully differentiated into osteoblasts,adipocytes,and chondrocytes in vitro.(2)The adipose-derived mesenchymal stem cells-derived exosomes met the identification criteria in terms of particle size,electron microscopy morphology,and positive expression of specific markers.(3)Compared to the dexamethasone-induced C2C12 atrophy group,treatment with adipose-derived mesenchymal stem cells and their exosomes promoted the recovery and growth of myotubes,inhibited the expression of muscle atrophy-related genes MuRF1 and Atrogin-1.(4)Compared to the aging C2C12 group,adipose-derived mesenchymal stem cells and their exosomes significantly enhanced the recovery and growth of aged muscle tubes in aging cells.(5)Compared to the control group,the rotarod time in dexamethasone-induced sarcopenia model mice was significantly decreased(P＜0.01).After 7 days(P＜0.01,P＜0.01)and 10 days(P＜0.01,P＜0.05)of adipose-derived mesenchymal stem cells treatment via intraperitoneal and intravenous injection,rotarod time was significantly increased,respectively.After 14 days,all treatment groups showed longer rotarod times than the model group,although with no significant differences between them.(6)Compared to the control group,the cross-sectional area of anterior tibial muscle in the model group was significantly reduced(P＜0.01),and it was significantly increased after intraperitoneal and intravenous administration of adipose-derived mesenchymal stem cells(P＜0.05,P＜0.01).(7)Compared to the model group,intraperitoneal and intravenous administration of adipose-derived mesenchymal stem cells significantly inhibited the mRNA expression of MuRF1 and Atrogin-1 genes(P＜0.01,P＜0.01,P＜0.01,P＜0.01).The results indicated that adipose-derived mesenchymal stem cells and their exosomes promoted recovery and growth of atrophic myotube cells by inhibiting the expression of muscle atrophy-related genes,and both intraperitoneal and intravenous administration of adipose-derived mesenchymal stem cells provided good therapeutic effects on sarcopenia in mice.

BACKGROUND:Sarcopenia is an age-related condition characterized by the loss of skeletal muscle mass,strength,and/or physical function.Currently,effective treatments for sarcopenia remain limited.A new therapeutic approach to improve symptoms and prognosis of sarcopenia patients clinically was important.OBJECTIVE:To explore the effects of canine adipose-derived mesenchymal stem cells and their exosomes on a dexamethasone-induced sarcopenia in mice.METHODS:Mesenchymal stem cells were isolated and cultured from canine adipose tissue,and identified and functionally evaluated through flow cytometry and differentiation assays for osteogenesis,adipogenesis,and chondrogenesis.Subsequently,exosomes from adipose-derived mesenchymal stem cells were extracted and characterized using transmission electron microscopy,western blot assay,and nanocoulter tracking analysis.In vitro,the effects of canine adipose-derived mesenchymal stem cells and their exosomes on myotube growth and the expression of muscle atrophy-related genes were investigated using dexamethasone-induced C2C12 myotube atrophy and aging C2C12 models.In vivo,a dexamethasone-induced mouse sarcopenia model was established and received intraperitoneal or intravenous injection of canine adipose-derived mesenchymal stem cells.Therapeutic efficacy was assessed through mouse rotarod performance,histopathological analysis,and muscle atrophy-related genes testing.RESULTS AND CONCLUSION:(1)The isolated canine adipose-derived mesenchymal stem cells highly expressed CD73,CD90,and CD105,and lowly expressed MHC-Ⅱ,CD14,CD19,CD34,and CD45,and successfully differentiated into osteoblasts,adipocytes,and chondrocytes in vitro.(2)The adipose-derived mesenchymal stem cells-derived exosomes met the identification criteria in terms of particle size,electron microscopy morphology,and positive expression of specific markers.(3)Compared to the dexamethasone-induced C2C12 atrophy group,treatment with adipose-derived mesenchymal stem cells and their exosomes promoted the recovery and growth of myotubes,inhibited the expression of muscle atrophy-related genes MuRF1 and Atrogin-1.(4)Compared to the aging C2C12 group,adipose-derived mesenchymal stem cells and their exosomes significantly enhanced the recovery and growth of aged muscle tubes in aging cells.(5)Compared to the control group,the rotarod time in dexamethasone-induced sarcopenia model mice was significantly decreased(P＜0.01).After 7 days(P＜0.01,P＜0.01)and 10 days(P＜0.01,P＜0.05)of adipose-derived mesenchymal stem cells treatment via intraperitoneal and intravenous injection,rotarod time was significantly increased,respectively.After 14 days,all treatment groups showed longer rotarod times than the model group,although with no significant differences between them.(6)Compared to the control group,the cross-sectional area of anterior tibial muscle in the model group was significantly reduced(P＜0.01),and it was significantly increased after intraperitoneal and intravenous administration of adipose-derived mesenchymal stem cells(P＜0.05,P＜0.01).(7)Compared to the model group,intraperitoneal and intravenous administration of adipose-derived mesenchymal stem cells significantly inhibited the mRNA expression of MuRF1 and Atrogin-1 genes(P＜0.01,P＜0.01,P＜0.01,P＜0.01).The results indicated that adipose-derived mesenchymal stem cells and their exosomes promoted recovery and growth of atrophic myotube cells by inhibiting the expression of muscle atrophy-related genes,and both intraperitoneal and intravenous administration of adipose-derived mesenchymal stem cells provided good therapeutic effects on sarcopenia in mice.

AIM: To explore the risk factors for visual field defect in patients with normal tension glaucoma.METHODS: The general data of 164 patients(164 eyes)with normal tension glaucoma diagnosed and treated in our hospital from July 2022 to January 2025 were retrospectively analyzed. According to the visual field defect, the patients were divided into visual field defect group(93 eyes)and no visual field defect group(71 eyes). The clinical data of the two groups were compared, and the risk factors of visual field defect in patients with normal intraocular pressure glaucoma were analyzed by Logistic regression.RESULTS: Totally 93 eyes had visual field defect among 164 eyes with normal tension glaucoma, with an incidence rate of 57%. The age, proportion of high myopia, intraocular pressure, intraocular pressure fluctuation and resistance index(RI)in the visual field defect group were higher than those in the non-visual field defect group(all P<0.01), while the retinal nerve fiber layer thickness, the optic disc vascular density, peak systolic blood flow velocity(PSV)and end diastolic blood flow velocity(EDV)in the visual field defect group were lower than those in the non-visual field defect group(all P<0.01). Multivariate Logistic regression showed that advanced age(OR=1.171), high myopia(OR=2.316), ocular hypertension(OR=1.662), high intraocular pressure fluctuation(OR=1.770), low retinal nerve fiber layer thickness(OR=0.744), low optic disc vascular density(OR=0.547), low PSV(OR=0.618), low EDV(OR=0.577)and high RI(OR=1.980)were all risk factors for visual field defect in patients with normal tension glaucoma(all P<0.01).CONCLUSION: The age, high myopia, intraocular pressure, intraocular pressure fluctuation, retinal nerve fiber layer thickness, optic disc vascular density, PSV, EDV and RI are influence factors for visual field defect in patients with normal tension glaucoma.

Objective To investigate the role of the ROS/NLRP3/Caspase-1 pyroptosis signaling pathway in mediating the inflammatory response of nerve roots in a rabbit model of lumbar disc herniation(LDH)following acupotomy intervention.Methods Forty New Zealand white rabbits were randomly assigned to four groups:a blank control group(10 rabbits),a model control group(10 rabbits),an electroacupuncture intervention group(positive control group,10 rabbits),and an acupotomy intervention group(10 rabbits).The LDH nerve root inflammatory response model was established in the model control group,electroacupuncture intervention group,and acupotomy intervention group using the classical autologous nucleus pulposus transplantation method.Following successful model establishment,corresponding interventions were administered.Dorsal root ganglion tissues were then collected for analysis.Reactive oxygen species levels were assessed using fluorescent probe staining.The expression of NLRP3 inflammasome-related proteins(NLRP3,ASC,GSDMD-N,and Caspase-1)was evaluated by Western blot.Mitochondrial permeability transition pore opening was assessed via Calcein AM staining.Results The comparison of relative fluorescence intensity of reactive oxygen species in dorsal root ganglion tissue among the groups indicated that the model control group exhibited significantly higher levels compared to the blank control group(P＜0.05).Both the electroacupuncture intervention group and the acupotomy intervention group demon-strated significantly lower levels than the model control group(P＜0.05).Western blot analysis revealed that the expression levels of NLRP3,ASC,GSDMD-N,and Caspase-1 in the model control group were significantly elevated compared to those in the blank control group(P＜0.05).Both intervention groups showed markedly reduced expression of these proteins compared to the model control group(P＜0.05).Additionally,the assessment of mito-chondrial permeability transition pore activity based on relative fluorescence intensity showed a significant increase in the model control group compared to the blank control group(P＜0.05),with both intervention groups exhibiting significantly lower levels than the model control group(P＜0.05).Conclusions Acupotomy therapy modulates the ROS/NLRP3/Caspase-1 signaling pathway,thereby inhibiting pyroptosis and attenuating the inflammatory response in nerve roots.This elucidates the potential molecular mechanism underlying the therapeutic effects of acupotomy on LDH through the regulation of ROS/NLRP3/Caspase-1-mediated downstream signaling cascades.

BACKGROUND:Scholars at home and abroad consider New Zealand rabbits to be an ideal model animal because of the similar anatomical morphology of the lumbar spine to that of the human lumbar spine.There is a lack of systematic comparison of different ways to establish rabbit intervertebral disc degeneration models under X-ray guidance. OBJECTIVE:To establish a rabbit model of lumbar disc degeneration using X-ray guided acupuncture,end-plate injection and combined method,and to compare the modeling effects of these three methods. METHODS:Eighteen 6-month-old New Zealand white rabbits were randomly selected and divided into four groups:acupuncture group,endplate injection group,combined group and blank control group.In the acupuncture group,three consecutive segments of the intervertebral discs(L2/3,L3/4,L4/5)were needled and modeled;in the endplate injection group,50 μL of anhydrous ethanol was injected at a single point on the endplates of the three consecutive segmental discs;in the combined group,three consecutive segmental intervertebral discs were needled and injected with 50 μL of anhydrous ethanol at four azimuthal points on the endplates of the corresponding segmental discs;and the blank control group received no interventions.X-ray examination was performed to measure the disc height index at 2,4,and 8 weeks after surgery.The intervertebral disc tissues were then taken for anatomical observation and pathological examination. RESULTS AND CONCLUSION:(1)Anatomical examination showed that fibrous annulus rupture,nucleus pulposus degeneration,and total disc structure disorder were the main manifestation in the acupuncture group,endplate injection group,and combined group,respectively.(2)X-ray examination showed that the disc height index showed the most obvious reduction in the acupuncture group at 2 weeks after operation,significant reductions in the endplate injection group at 2 and 4 weeks after operation,and significant reductions in the combined group at 2,4,and 8 weeks after operation.(3)Pathological examination showed that the fibrous ring structure was damaged and the inner annulus fibrosus protruded inward in the acupuncture group;endplate fissure,disordered arrangement and nucleus loss were observed in the endplate injection group;total disc structure disorder with the nucleus pulposus losing water and shrinking and no obvious border with the broken annulus fibrosus was found in the combined group.To conclude,acupuncture,endplate injection and the modified endplate injection method can establish the rabbit intervertebral disc degeneration model.Compared with the single method,the modified endplate injection method can greatly accelerate and aggravate the degeneration of the intervertebral disc,and can effectively shorten the experiment period.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Objective To investigate the role of the ROS/NLRP3/Caspase-1 pyroptosis signaling pathway in mediating the inflammatory response of nerve roots in a rabbit model of lumbar disc herniation(LDH)following acupotomy intervention.Methods Forty New Zealand white rabbits were randomly assigned to four groups:a blank control group(10 rabbits),a model control group(10 rabbits),an electroacupuncture intervention group(positive control group,10 rabbits),and an acupotomy intervention group(10 rabbits).The LDH nerve root inflammatory response model was established in the model control group,electroacupuncture intervention group,and acupotomy intervention group using the classical autologous nucleus pulposus transplantation method.Following successful model establishment,corresponding interventions were administered.Dorsal root ganglion tissues were then collected for analysis.Reactive oxygen species levels were assessed using fluorescent probe staining.The expression of NLRP3 inflammasome-related proteins(NLRP3,ASC,GSDMD-N,and Caspase-1)was evaluated by Western blot.Mitochondrial permeability transition pore opening was assessed via Calcein AM staining.Results The comparison of relative fluorescence intensity of reactive oxygen species in dorsal root ganglion tissue among the groups indicated that the model control group exhibited significantly higher levels compared to the blank control group(P＜0.05).Both the electroacupuncture intervention group and the acupotomy intervention group demon-strated significantly lower levels than the model control group(P＜0.05).Western blot analysis revealed that the expression levels of NLRP3,ASC,GSDMD-N,and Caspase-1 in the model control group were significantly elevated compared to those in the blank control group(P＜0.05).Both intervention groups showed markedly reduced expression of these proteins compared to the model control group(P＜0.05).Additionally,the assessment of mito-chondrial permeability transition pore activity based on relative fluorescence intensity showed a significant increase in the model control group compared to the blank control group(P＜0.05),with both intervention groups exhibiting significantly lower levels than the model control group(P＜0.05).Conclusions Acupotomy therapy modulates the ROS/NLRP3/Caspase-1 signaling pathway,thereby inhibiting pyroptosis and attenuating the inflammatory response in nerve roots.This elucidates the potential molecular mechanism underlying the therapeutic effects of acupotomy on LDH through the regulation of ROS/NLRP3/Caspase-1-mediated downstream signaling cascades.

Objective:To construct classification prediction models for gamma passing rate using radiomics-based machine learning approaches and data from multiple radiotherapy institutions and evaluate the models′ performance.Methods:The data from 572 volumetric-modulated arc therapy (VMAT) patients across three radiotherapy institutions (514 for training and 58 for testing)were retrospectively collected. Additionally, 45 VMAT plans were collected from a single institution as an independent external validation set. For all the data, a three-dimensional dose validation approach based on actual measurements of phantoms was utilized, and gamma analysis was performed at the 3%/2 mm criterion using a dose threshold of 10%, absolute doses, and global normalization. After radiomic features were extracted from dose files, feature selection was performed using the random forest (RF) method and RF combined with Shapley Additive exPlanation (SHAP). Then, feature subsets of varying sizes (10, 20, 30, 40, and 50) were selected based on feature rankings. Using these subsets as inputs, data training was conducted using the Extreme Gradient Boosting (XGBoost) algorithm. Finally, the models′ classification performance was assessed using the area under the curve (AUC) values and F1-score.Results:Under the 3%/2 mm criterion, all models performed the best in the case of 20 feature subsets. The optimal prediction model established based on the feature selection using RF exhibited AUC and F1-score of 0.88 and 0.89, respectively on the testing set and 0.82 and 0.90, respectively, on the validation set. The optimal prediction model built based on the feature selection using RF combined with SHAP yielded AUC and F1-score of 0.86 and 0.92 on the testing set and 0.87 and 0.89, respectively, on the validation set, along with superior robustness. Therefore, the second model possessed certain advantages over the first model.Conclusions:For multicenter dose verification result, it is feasible to construct a machine learning prediction model with high classification performance using radiomic features derived from dose files, combined with feature selection based on SHAP. This approach can assist in advancing the clinical applications and implementation of gamma passing rate prediction models.

Objective Explore a non-contact physiological and psychological detection model based on facial video in simulations of weightlessness effects,research new methods for non-contact heart rate and negative mood state detection in long-term simulations of weightlessness effect analysis.Methods Construct a non-contact physiological and psychological data collection system for fusion analysis of visible light and thermal infrared videos.Collect physiological and psychological data of volunteers in the"Earth Star-Ⅱ"90-day head-down bed rest experiment.A non-contact heart rate detection model based on GCN facial multi-region feature fusion and a non-contact negative mood state detection model considering data reliability were constructed,and the effectiveness of the models were validated with finger clip heart rate and POMS-SF scale as labels.Results The experimental results show that the average difference in the Bland-Altman plot of the non-contact heart rate detection model is-1.26 bpm,and 96.3%of value error detection data falls within the 95%confidence interval,indicating a high consistency between the model detected heart rate and the finger clip heart rate.The non-contact negative mood state detection model achieves an accuracy of>0.85 for detecting tension,depression,anger,and fatigue.Features such as heart rate,AU06,eye gaze,and head pose were observed to be important to mood state detection.Conclusion Non-contact physiological and psychological detection methods not only can be utilized for long-term physiological analysis in simulations of weightlessness effects,but also provide a novel technical approach for on-orbit astronauts health assurance during long-term space flight in the future.