BACKGROUND:Human periodontal ligament stem cells are potential functional cells for periodontal tissue engineering.However,long-term in vitro culture may lead to reduced stemness and replicative senescence of periodontal ligament stem cells,which may impair the therapeutic effect of human periodontal ligament stem cells. OBJECTIVE:To investigate the effect of oxymatrine on the stemness maintenance and osteogenic differentiation of periodontal ligament stem cells in vitro,and to explore the potential mechanism. METHODS:Periodontal ligament stem cells were isolated from human periodontal ligament tissues by tissue explant enzyme digestion and cultured.The surface markers of mesenchymal cells were identified by flow cytometry.Periodontal ligament stem cells were incubated with 0,2.5,5,and 10 μg/mL oxymatrine.The effect of oxymatrine on the proliferation activity of periodontal ligament stem cells was detected by CCK8 assay.The appropriate drug concentration for subsequent experiments was screened.Western blot assay was used to detect the expression of stem cell non-specific proteins SOX2 and OCT4 in periodontal ligament stem cells.qRT-PCR and western blot assay were used to detect the expression levels of related osteogenic genes and proteins in periodontal ligament stem cells. RESULTS AND CONCLUSION:(1)The results of CCK8 assay showed that 2.5 μg/mL oxymatrine significantly enhanced the proliferative activity of periodontal stem cells,and the subsequent experiment selected 2.5 μg/mL oxymatrine to intervene.(2)Compared with the blank control group,the protein expression level of SOX2,a stem marker of periodontal ligament stem cells in the oxymatrine group did not change significantly(P>0.05),and the expression of OCT4 was significantly up-regulated(P<0.05).(3)Compared with the osteogenic induction group,the osteogenic genes ALP,RUNX2 mRNA expression and their osteogenic associated protein ALP protein expression of periodontal ligament stem cells were significantly down-regulated in the oxymatrine+osteogenic induction group(P<0.05).(4)The oxymatrine up-regulated the expression of stemness markers of periodontal ligament stem cells and inhibited the bone differentiation of periodontal ligament stem cells,and the results of high-throughput sequencing showed that it may be associated with WNT2,WNT16,COMP,and BMP6.

AIM:To investigate the effects of electroacupuncture(EA)at"Qihai","Zhongji"and"Guanyuan"acupoints on urinary control function in rats with stress urinary incontinence(SUI)as well as to explore the mechanism of regulating ferroptosis.METHODS:A total of 50 Sprague-Dawley(SD)rats were randomly assigned to 5 groups.The rats in control(Con)group did not receive any treatment.The rats in model group was modeled for bilateral oophorectomy and vaginal dilation.The rats in EA group received EA treatment at"Qihai","Zhongji"and"Guanyuan"acupoints after modeling,while those in sham acupuncture(sham)group received EA treatment beside the specified acupoints after modeling.The rats in EA+solute carrier family 7 member 11(SLC7A11)inhibitor imidazole ketone erastin(IKE)group received an intraperitoneal injection of 50 mg/kg IKE and EA intervention after modeling.After the experiment,the uri-nary control indicators of leak point pressure(LPP),abdominal LPP(ALPP),and maximum bladder capacity(MBC)were measured for all rats.Hematoxylin-eosin(HE),Masson and Prussian blue staining was performed to observe the pathological changes and iron deposition in the urethral sphincter muscle.Enzyme-linked immunosorbent assay(ELISA)was performed to detect the mitochondrial functional indicators(mitochondrial membrane potential and ATP)and redox in-dicators:changes in the levels of reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH).RT-qPCR,Western blot and immunofluorescence staining were performed to detect the expression of key iron death proteins.RESULTS:When compared with the Con group,the main urinary control indicators of MBC,LPP,and ALPP in the model group were significantly reduced(P<0.05),and the urethral sphincter fibers were broken and disordered,iron de-position was significant,and the proportion of muscle fibers was significantly reduced.The mitochondrial function indica-tors and GSH levels were significantly reduced(P<0.05),while the ROS intensity,Fe2+and MDA levels were significant-ly increased(P<0.05),the glutathione peroxidase 4(GPX4)and SLC7A11 mRNA protein expressions were significantly reduced(P<0.05),and the acyl-CoA synthetase long-chain family member 4(ACSL4)and transferrin receptor 1(TFR1)protein expressions were significantly increased(P<0.05).When compared with the model group,the main urinary con-trol indicators of the rats in the EA group were significantly improved(P<0.05).The urethral sphincter fibers breakage and disordered were attenuated,iron deposition was significantly reduced,and the proportion of muscle fibers was signifi-cantly increased.The mitochondrial function indicators and GSH levels were significantly increased(P<0.05),while the ROS intensity,Fe2+and MDA levels were significantly reduced(P<0.05).The mRNA and protein expressions of GPX4 and SLC7A11 were significantly increased(P<0.05),and the mRNA and protein expressions of ACSL4 and TFR1 were significantly reduced(P<0.05).When compared with the EA group,the above indicators in the EA+IKE group showed negative changes,and the difference was statistically significant(P<0.05).CONCLUSION:Treatment with EA at"Qi-hai","Zhongji"and"Guanyuan"acupoints improves urinary function in SUI,and the underlying mechanism may be relat-ed to ferroptosis mediated by the SLC7A11/GPX4 signaling pathway.

Objective To explore whether creatine therapy regulates neuronal ferroptosis by inhibiting the activation of STAT1 signaling pathway associated with suppressor of cytokine signaling 1(SOCS1)in Alzheimer's disease.Methods Immunohistochemical staining and counting of positive results using paraffin sections of human brain frontal lobes were employed to determine the trend of changes in the target proteins.Further validation was performed by immunofluorescence and Western blotting.STAT1 phosphorylation was inhibited by creatine injection using eleven FAD4T mice and by cerebellar medullary pool puncture,and the expression of target proteins was examined by immunohistochemistry and immunofluorescence after postmortem sampling.Results Compared with the age controls,interferon-γ(IFN-γ),an activating cytokine of the STAT1 signaling pathway,and SOCS1,a negative regulator of STAT1 activation,were both significantly up-regulated,STAT1 phosphorylation was enhanced,and the ferroptosis markers ferritin light chain(FTL)and cystine/glutamate transporter(xCT)increased markedly in the cortex of AD human brains;Creatine treatment of FAD4T mice resulted in a reduction of both IFN-γ and SOCS1,and a significant decrease in the ferroptosis markers FTL and xCT(SLC7A11).Conclusion Creatine ameliorates neuronal ferroptosis in AD model mice by reducing neuronal STAT1-SOCS1 signalling activation.

The orientation of the acetabular cup in hip joint anteroposterior radiograph is a key factor in evaluating the postoperative outcomes of total hip arthroplasty (THA). Currently, measurement of the acetabular cup anteversion angle primarily relies on manual drawing of auxiliary lines by orthopedic surgeons and calculations using scientific calculators. This study proposes an automated computer-aided measurement method for the acetabular cup anteversion angle based on hip joint anteroposterior radiograph. The proposed method segments hip prosthesis images using an improved Otsu algorithm, identifies feature points at the acetabular cup opening by combining circle-fitting theory and the cup's geometric characteristics, and fits an ellipse to the cup opening to calculate the anteversion angle. A total of 104 hip joint anteroposterior radiographs, including 71 right-sided and 81 left-sided prostheses, were analyzed. Two orthopedic surgeons independently measured the postoperative anteversion angles, and the results were compared with computer-generated measurements for correlation analysis. Spearman and Pearson correlation analyses demonstrated significant correlations between the proposed method and manual measurements for both the right group ( r = 0.795, P < 0.01) and the left group ( r = 0.859, P < 0.01). This method provides a reliable reference for orthopedic surgeons to assess postoperative prognosis.
Humans ; Acetabulum/anatomy & histology* ; Arthroplasty, Replacement, Hip/methods* ; Algorithms ; Hip Prosthesis ; Hip Joint/diagnostic imaging* ; Radiography ; Image Processing, Computer-Assisted/methods*

Acute lateral ankle sprain (ALAS) is one of the most common sport injuries, with high incidence, recurrence and disability rates. Currently, exercise rehabilitation-based non-surgical treatment is the primary management approach for ALAS. However, there remain improper practices such as excessive immobilization or uncontrolled activity, which contribute to recurrent sprains and chronic ankle instability, significantly impairing patients′ athletic function and quality of life. To standardize the non-surgical management of ALAS, improve the cure rates, and reduce the recurrence and disability rates, Chinese Sports Rehabilitation Medicine Training Project of Chinese Medical Association, Foot and Ankle Basics and Orthopedics Group, Orthopedic Branch of Chinese Medical Doctor Association, and Sports Medicine Branch of Jiangsu Medical Association organized relevant experts to formulate Expert consensus on non-surgical treatment for acute lateral ankle sprain ( version 2025), following the principles of scientific vigor, practicality, and innovation. Thirteen recommendations were proposed for standardized treatment protocols across different healing phases, aiming to provide references for standard management of ALAS and improve the therapeutic outcomes.

Thoracolumbar spine fracture often leads to severe pain, functional impairments, and neurological deficits, for which open reduction and internal fixation can effectively restore the spinal structural stability. Open decompression and reduction with internal fixation can help relieve spinal cord compression and improve spinal function in cases of concomitant cord injury. Although spinal stability can be restored through surgery, patients often face chronic pain and functional impairments postoperatively. A postoperative rehabilitation program is critical in optimizing therapeutic outcomes, reducing complications, and minimizing the risk of secondary injuries. However, current rehabilitation methods, such as physical therapy, functional training, and pain management, are confronted with problems in clinical practice, including significant variation in efficacy, poor patient adherence, and prolonged rehabilitation period. There is an urgent need for a unified rehabilitation strategy to address these problems. To this end, the Spinal Trauma Group of the Orthopedic Physicians Branch of the Chinese Medical Association and the Spine Health Professional Committee of the Chinese Human Health Technology Promotion Association organized experts from relevant fields to formulate Evidence-based guidelines for rehabilitation treatment after internal fixation of thoracolumbar spine fracture in adults ( version 2025) by integrating evidences from clinical researches and advanced rehabilitation concepts at home and abroad. A total number of 14 recommendations concerning the rehabilitation treatment with multimodal analgesia, psychological intervention, deep vein thrombosis prevention, core muscle and extremity exercise, appropriate use of braces, early weight-bearing, device-aided rehabilitation exercise, neuroregulatory therapy, rehabilitation team were put forward, aiming to standardize the post-operative rehabilitation process following internal fixation, promote the functional recovery, and enhance patients′ quality of life.

Lentiviral vectors(LVs)are key gene delivery tools for integrating target genes into the host genome,but they may also pose risks of insertional mutagenesis.The vector copy number(VCN)in cells is critical for determining the safety of gene modification.However,the reliability and accuracy of its quantification process are influenced by multiple factors.Developing cell reference materials with specific vector copy numbers represents a viable approach to enhance the reliability and consistency of measurement results,enabling quality control of the quantification process and traceability of outcomes.However,the preparation of such reference materials faces challenges in cell sample design,preparation protocols,and advanced quantification techniques.In this study,T lymphocyte cell line Jurkat-based reference materials with LV gene copy numbers of 1 and 2 copy/cell were developed.A high-precision duplex digital polymerase chain reaction(dPCR)method was established to quantify the LV gene and endogenous genes simultaneously.Additionally,the results of dPCR were cross-validated through next-generation sequencing and flow cytometric analysis.Ultimately,confocal microscopy characterization results showed that the developed cell reference materials had intact morphology.The quantification result of VCN-1 was(1.07±0.11)copy/cell,and that of VCN-2 was(2.09±0.21)copy/cell.These cell reference materials demonstrated compliance with stability and homogeneity requirements,and could be applied for quality control throughout the VCN measurement workflow and metrological traceability,improving the accuracy,comparability,and validity of copy number measurements.

Objective:To evaluate the efficacy of preoperative prediction of regional lymph node (RLN) metastasis in renal cell carcinoma (RCC) using a machine learning model based on habitat imaging radiomics from renal MRI.Methods:This cross-sectional study retrospectively analyzed 220 patients with RCC who underwent nephrectomy and RLN dissection at four medical centers of Chinese PLA General Hospital from January 2010 to August 2023. The cohort included 65 patients with RLN metastasis and 155 without. A stratified random sampling method was used to divide 175 patients from the first medical center into a training set ( n=140) and an internal test set ( n=35) in an 8∶2 ratio, while 45 patients from the third, fourth, and fifth medical centers constituted the external test set. The primary RCC lesions were categorized into 15 habitat subregions based on corticomedullary-phase enhancement and T 2WI signal intensity on MRI, and the volume fractions of different subregions were analyzed. In the training cohort, radiomics features derived from the habitat subregions were used to construct a radiomics model employing various machine learning algorithms, including extremely random trees (ET), gradient boosting decision trees (GBDT), random forest (RF), and support vector machine (SVM). The optimal model was selected and combined with RLN short-axis diameter to develop a combined model. The efficacy of each model in predicting RLN metastasis was evaluated using the receiver operating characteristic (ROC) curve. Results:The volume fraction of hyper-enhanced hyper-intense regions in the non-metastatic group was significantly higher than that in the metastatic group (0.05±0.09 vs. 0.02±0.03; t=3.00, P=0.003). Among the machine learning models constructed using 15 optimal habitat radiomics features, the SVM model demonstrated the best performance, with area under the ROC curve (AUC) values of 0.85 (95% CI 0.72-0.98) in the internal test set and 0.82 (95% CI 0.67-0.98) in the external test set, surpassing those of the ET, GBDT, and RF models. The combined model, integrating the SVM model with RLN short-axis diameter, achieved AUC values of 0.94 (95% CI 0.85-1.00) in the internal test set and 0.89 (95% CI 0.78-1.00) in the external test set, with RLN short-axis diameter contributing AUC values of 0.81 (95% CI 0.66-0.96) and 0.81 (95% CI 0.68-0.94), respectively. The diagnostic sensitivity of the combined model was 91.7% in the internal test set and 85.7% in the external test set, with specificities of 78.3% and 67.7%, respectively. Conclusion:The combined model based on MRI habitat imaging radiomics and RLN short-axis diameter demonstrates excellent preoperative assessment capability for RLN metastasis in RCC.

AIM:To investigate the effects of electroacupuncture(EA)at"Qihai","Zhongji"and"Guanyuan"acupoints on urinary control function in rats with stress urinary incontinence(SUI)as well as to explore the mechanism of regulating ferroptosis.METHODS:A total of 50 Sprague-Dawley(SD)rats were randomly assigned to 5 groups.The rats in control(Con)group did not receive any treatment.The rats in model group was modeled for bilateral oophorectomy and vaginal dilation.The rats in EA group received EA treatment at"Qihai","Zhongji"and"Guanyuan"acupoints after modeling,while those in sham acupuncture(sham)group received EA treatment beside the specified acupoints after modeling.The rats in EA+solute carrier family 7 member 11(SLC7A11)inhibitor imidazole ketone erastin(IKE)group received an intraperitoneal injection of 50 mg/kg IKE and EA intervention after modeling.After the experiment,the uri-nary control indicators of leak point pressure(LPP),abdominal LPP(ALPP),and maximum bladder capacity(MBC)were measured for all rats.Hematoxylin-eosin(HE),Masson and Prussian blue staining was performed to observe the pathological changes and iron deposition in the urethral sphincter muscle.Enzyme-linked immunosorbent assay(ELISA)was performed to detect the mitochondrial functional indicators(mitochondrial membrane potential and ATP)and redox in-dicators:changes in the levels of reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH).RT-qPCR,Western blot and immunofluorescence staining were performed to detect the expression of key iron death proteins.RESULTS:When compared with the Con group,the main urinary control indicators of MBC,LPP,and ALPP in the model group were significantly reduced(P<0.05),and the urethral sphincter fibers were broken and disordered,iron de-position was significant,and the proportion of muscle fibers was significantly reduced.The mitochondrial function indica-tors and GSH levels were significantly reduced(P<0.05),while the ROS intensity,Fe2+and MDA levels were significant-ly increased(P<0.05),the glutathione peroxidase 4(GPX4)and SLC7A11 mRNA protein expressions were significantly reduced(P<0.05),and the acyl-CoA synthetase long-chain family member 4(ACSL4)and transferrin receptor 1(TFR1)protein expressions were significantly increased(P<0.05).When compared with the model group,the main urinary con-trol indicators of the rats in the EA group were significantly improved(P<0.05).The urethral sphincter fibers breakage and disordered were attenuated,iron deposition was significantly reduced,and the proportion of muscle fibers was signifi-cantly increased.The mitochondrial function indicators and GSH levels were significantly increased(P<0.05),while the ROS intensity,Fe2+and MDA levels were significantly reduced(P<0.05).The mRNA and protein expressions of GPX4 and SLC7A11 were significantly increased(P<0.05),and the mRNA and protein expressions of ACSL4 and TFR1 were significantly reduced(P<0.05).When compared with the EA group,the above indicators in the EA+IKE group showed negative changes,and the difference was statistically significant(P<0.05).CONCLUSION:Treatment with EA at"Qi-hai","Zhongji"and"Guanyuan"acupoints improves urinary function in SUI,and the underlying mechanism may be relat-ed to ferroptosis mediated by the SLC7A11/GPX4 signaling pathway.

Acute lateral ankle sprain (ALAS) is one of the most common sport injuries, with high incidence, recurrence and disability rates. Currently, exercise rehabilitation-based non-surgical treatment is the primary management approach for ALAS. However, there remain improper practices such as excessive immobilization or uncontrolled activity, which contribute to recurrent sprains and chronic ankle instability, significantly impairing patients′ athletic function and quality of life. To standardize the non-surgical management of ALAS, improve the cure rates, and reduce the recurrence and disability rates, Chinese Sports Rehabilitation Medicine Training Project of Chinese Medical Association, Foot and Ankle Basics and Orthopedics Group, Orthopedic Branch of Chinese Medical Doctor Association, and Sports Medicine Branch of Jiangsu Medical Association organized relevant experts to formulate Expert consensus on non-surgical treatment for acute lateral ankle sprain ( version 2025), following the principles of scientific vigor, practicality, and innovation. Thirteen recommendations were proposed for standardized treatment protocols across different healing phases, aiming to provide references for standard management of ALAS and improve the therapeutic outcomes.