BACKGROUND:Acupoint embedding and floating needle therapy are two methods for the treatment of knee osteoarthritis,but there are few reports on the combined treatment of the two methods. OBJECTIVE:To investigate the therapeutic effect of acupoint embedding combined with floating needle therapy on different stages of knee osteoarthritis using generalized estimating equations. METHODS:A total of 436 patients with knee osteoarthritis admitted to Dongguan Hospital of Traditional Chinese Medicine from February 2019 to February 2023 were selected as the research subjects.All patients were randomly divided into a control group with floating needle therapy(n=218)and an observation group with acupoint embedding method combined with floating needle therapy(n=218).Staging was performed according to the K-L staging method.In the control group,there were 57 cases in stage Ⅰ,62 in stage Ⅱ,49 in phase Ⅲ,and 50 in stage Ⅳ,while in the observation group,there were 48 cases in stage Ⅰ,66 in stage Ⅱ,63 in phase Ⅲ,and 41 in stage Ⅳ.The levels of indexes and clinical efficacy were compared between groups before and after treatment.Generalized estimating equation model was used to analyze the influencing factors of clinical efficacy and the interaction effect of different time points,different methods and different stages on therapeutic efficacy. RESULTS AND CONCLUSION:There was no significant difference in baseline data between the observation group and the control group,as well as between the patients of different stages(P>0.05).After treatment,the cure rate of stage Ⅰ patients was the highest after treatment,and the total improvement rate in the observation group was significantly higher than that in the control group.There were significant differences in the cure rate among different stages in each group(P<0.05).After treatment,all indicators in the two groups were significantly decreased.In the control group,there were significant differences in various indicators of patients at different stages after 4 weeks of treatment(P<0.05).In the observation group,after 2 weeks of treatment,there were significant differences in various indicators of patients at different stages(P<0.05),and all the indexes in the observation were lower than those in the control group after 4 weeks of treatment(P<0.05)and the therapeutic effect in the observation group was better than that in the control group.Generalized estimating equation model analysis showed that trauma history,interleukin-6 level,treatment method,treatment time and K-L stage all significantly affected the clinical efficacy in patients.In the interaction effect analysis,after 2 weeks of treatment,there was a significant difference in the visual analogue scale score between the two groups in stages Ⅲ and Ⅳ(P<0.05).After 4 weeks of treatment,there was a significant difference in the visual analogue scale score between the two groups in stages Ⅱ,Ⅲ,and Ⅳ(P<0.05).To conclude,acupoint embedding combined with floating needle therapy is superior to floating needle therapy alone in the treatment of different stages of knee osteoarthritis.Trauma history,interleukin-6 level,treatment method,treatment time and K-L stage significantly influence the therapeutic effect.

BACKGROUND:The diabetic microenvironment can cause excessive M1 polarization of macrophages,and this hyperglycemic inflammatory state can inhibit osteogenic differentiation of bone marrow mesenchymal stem cells,thus affecting the healing of diabetic bone defects.Studies have indicated that mogroside V possesses anti-inflammatory,antioxidant,and hypoglycemic properties.However,its potential to modulate M1 polarization of macrophages and osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition remains unclear. OBJECTIVE:To explore the effect of mogroside V on regulating M1 macrophage polarization and its effect on osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition. METHODS:Murine diabetic models were established using C57BL/6 mice.Bone marrow-derived macrophages were isolated from tibia and fibula of normal and diabetic mice,and cultured in low-glucose and high-glucose media.Then M1 polarization of bone marrow-derived macrophages was induced using lipopolysaccharide and interferon-γ.Bone marrow-derived macrophages were treated with 160,320,and 640 μmol/L mogroside V.Flow cytometry was employed to determine the proportion of F4/80+CD86+cells.qRT-PCR was utilized to assess mRNA expression levels of inducible nitric oxide synthase,interleukin 1β,and interleukin 6.ELISA was employed to evaluate tumor necrosis factor-α secretion in bone marrow-derived macrophage supernatants.Bone marrow mesenchymal stem cells were isolated from tibia and fibula of C57BL/6 suckling mice,and induced osteogenic differentiation using low-or high-glucose osteogenic induction medium.Bone marrow mesenchymal stem cells were treated with M1 macrophage-conditioned mediums with or without 320 μmol/L mogroside V in osteogenic differentiation process.qRT-PCR was employed to assess the mRNA expression of alkaline phosphatase,Runt-related factor 2,osteocalcin,and osteopontin on day 14 after osteogenic induction.Alizarin red staining and quantitative analysis were conducted to evaluate calcium deposition on day 21 after osteogenic induction. RESULTS AND CONCLUSION:(1)Flow cytometry results showed that with the treatment of 320 and 640 μmol/L mogroside V,the proportion of F4/80+CD86+bone marrow-derived macrophages was significantly lower than that in the high-glucose control group(P<0.05).(2)qRT-PCR results showed that with the treatment of 160,320,and 640 μmol/L mogroside V,the mRNA expression levels of inducible nitric oxide synthase and interleukin 6 were significantly lower than that in the high-glucose control group(P<0.05).With the treatment of 320 and 640 μmol/L mogroside V,the mRNA expression level of interleukin 1β was significantly lower than that in the high-glucose control group(P<0.05).(3)ELISA results exhibited that with the treatment of 160,320,and 640 μmol/L mogroside V,the tumor necrosis factor-α secretion level was significantly lower than that in the high-glucose control group(P<0.05).(4)With the treatment of 320 μmol/L mogroside V,calcium salt deposition was increased in bone marrow mesenchymal stem cells under high glucose and inflammatory conditions(P<0.05),and the mRNA relative expression levels of alkaline phosphatase,Runt-related factor 2,and osteopontin were increased(P<0.05).These findings indicate that mogroside V can promote osteogenic differentiation of bone marrow mesenchymal stem cells by inhibiting the M1 polarization of bone marrow-derived macrophages under high glucose and inflammatory conditions and reducing the generation of inflammatory factors.

BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.

ObjectiveTobacco-related diseases remain one of the leading preventable public health challenges worldwide and are among the primary causes of premature death. In recent years, accumulating evidence has supported the classification of nicotine addiction as a chronic brain disease, profoundly affecting both brain structure and function. Despite the urgency, effective diagnostic methods for smoking addiction remain lacking, posing significant challenges for early intervention and treatment. To address this issue and gain deeper insights into the neural mechanisms underlying nicotine dependence, this study proposes a novel graph neural network framework, termed TI-GNN. This model leverages functional magnetic resonance imaging (fMRI) data to identify complex and subtle abnormalities in brain connectivity patterns associated with smoking addiction. MethodsThe study utilizes fMRI data to construct functional connectivity matrices that represent interaction patterns among brain regions. These matrices are interpreted as graphs, where brain regions are nodes and the strength of functional connectivity between them serves as edges. The proposed TI-GNN model integrates a Transformer module to effectively capture global interactions across the entire brain network, enabling a comprehensive understanding of high-level connectivity patterns. Additionally, a spatial attention mechanism is employed to selectively focus on informative inter-regional connections while filtering out irrelevant or noisy features. This design enhances the model’s ability to learn meaningful neural representations crucial for classification tasks. A key innovation of TI-GNN lies in its built-in causal interpretation module, which aims to infer directional and potentially causal relationships among brain regions. This not only improves predictive performance but also enhances model interpretability—an essential attribute for clinical applications. The identification of causal links provides valuable insights into the neuropathological basis of addiction and contributes to the development of biologically plausible and trustworthy diagnostic tools. ResultsExperimental results demonstrate that the TI-GNN model achieves superior classification performance on the smoking addiction dataset, outperforming several state-of-the-art baseline models. Specifically, TI-GNN attains an accuracy of 0.91, an F1-score of 0.91, and a Matthews correlation coefficient (MCC) of 0.83, indicating strong robustness and reliability. Beyond performance metrics, TI-GNN identifies critical abnormal connectivity patterns in several brain regions implicated in addiction. Notably, it highlights dysregulations in the amygdala and the anterior cingulate cortex, consistent with prior clinical and neuroimaging findings. These regions are well known for their roles in emotional regulation, reward processing, and impulse control—functions that are frequently disrupted in nicotine dependence. ConclusionThe TI-GNN framework offers a powerful and interpretable tool for the objective diagnosis of smoking addiction. By integrating advanced graph learning techniques with causal inference capabilities, the model not only achieves high diagnostic accuracy but also elucidates the neurobiological underpinnings of addiction. The identification of specific abnormal brain networks and their causal interactions deepens our understanding of addiction pathophysiology and lays the groundwork for developing targeted intervention strategies and personalized treatment approaches in the future.

Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.

Objective To analyze the detection rate of carbapenem-resistant Klebsiella pneumoniae(CRKP)in Guangdong Provincial Second Hospital of Traditional Chinese Medicine(the hospital)and analyze the main drug resistance genes and virulence genes of CRKP,so as to understand the molecular epidemiologi-cal mechanism of its infection strains.Methods The detection rate of CRKP infection in the hospital from 2020 to 2023 was retrospectively analyzed,and a total of 84 strains of CRKP were collected from July to De-cember 2022 in the hospital.The clinical data of the strains were collected,and the corresponding drug resist-ance genes and virulence genes were amplified by PCR.The modified carbapenem inactivation method(mCIM)was uesd to detect carbapenemase.Results The detection rates of CRKP in the Guangdong Provincial Second Hospital of Traditional Chinese Medicine from 2020 to 2023 were relatively high,and were higher than 46.00％.84 non-repeated CRKP strains were collected from July to December of 2022,and most samples were from respiratory tract,accounting for 55.95％.The patients were mainly from acupuncture and rehabilitation departments,accounting for 34.52％.Drug sensitivity tests showed that CRKP was highly resistant to various cephalosporin and extended spectrum beta lactam drugs,and only showed high sensitivity to tigecycline and polymyxin.The positive rate of mCIM test was 84.52％(71/84),and the other 15.48％results were neutral,which failed to determine whether they produced carbapenemase.A total of 73 strains were detected with car-bapenemase gene,accounting for 86.90％,involving 4 genotypes.The detection rates of blaKPC,blaNDM,blaIMP,and blaOXA-48 were 83.33％,2.38％,1.19％,and 1.19％,respectively.One of them carried both blaKPC and blaNDM genes,and multiple β-lactamases were detected.The detection rates of blaSHV,bla-TEM,blaCTX-M-9,and blaCTX-M-1 were 96.43％,78.57％,64.29％,and 2.38％,respectively.The detection rates of five virulence genes,blaiucA,blarmpA2,blairoB,blapeg-334,and blarmpA,were 42.86％,41.67％,27.38％,3.57％,and 2.38％,respectively.The proportion of strains carrying three or more virulence genes was 17.85％(15/84).Conclusion The detection rate of CRKP in the hospital is relatively high,and the drug resistance situation is serious,with Klebsiella pneumoniae carbapenemase(KPC)as the main drug resistance gene.A high proportion of Carbapenem-resistant hypervirulent Klebsiella pneumoniae(CR-hvKp)strains is found,and the infection prevention and control situation is not optimistic.It is necessary to further strengthen the hospital infection control measures and standardize the scientific and reasonable drug use.

NLRP3 can recruit proteins such as ASC and pro-caspase1 to form NLRP3 inflammasomes after being stimulated by pathogen and danger signals in vivo,and then induce pyropto-sis and promote the inflammatory reactions to maintain the home-ostasis.However,the overactivation of NLRP3 inflammasomes is closely related to many inflammatory and autoimmune diseases in humans.Targeted inhibition of NLRP3 inflammasomes can sig-nificantly inhibit inflammation and alleviate the relative symp-toms.Therefore,it is an important research direction for treating diseases of NLRP3 inflammasome that searching for effective in-hibitors targeting NLRP3 inflammasome activation and achieving clinical transformation.This review summarizes the latest re-search progress based on the sources of NLRP3 inflammasome inhibitors.

OBJECTIVE To investigate the protective effect of interleukin-35(IL-35)mRNA-lipid nanoparticles(LNP)against lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice.METHODS Fifity-six mice were randomly divided into 7 groups with 8 mice in each,including the normal control group,IL-35 mRNA-LNP(250 μg·kg-1)group,LPS group,LPS+IL-35 mRNA-LNP(50,125 and 250 μg·kg-1)group and LPS+Dexamethasone(DXM)group.Except for the normal control group and IL-35 mRNA-LNP(250 μg·kg-1)group and ALI model was established by tracheal infusion of LPS in each of the other groups.IL-35 mRNA-LNP(250 μg·kg-1)group and LPS+IL-35 mRNA-LNP(50,125 and 250 μg·kg-1)group were injected with a corresponding dose of LNP encapsulated mRNA complex via the tail vein while the LPS+DXM group was injected with DXM via the tail vein.Lung coefficient and the wet to dry weight ratio(W/D)of lung tissue were recorded.The mRNA levels of inflammatory cytokines tumor necrosis factor-α(TNF-α),Interleukin-6(IL-6)and IL-1βof lung homogenates were detected by real-time fluorescence quantitative PCR(RT-qPCR).LDH activity of lung homogenates and the protein levels of IL-35,TNF-α,IL-6 and IL-1β in lung homogenate were detected by corresponding kits.Hematoxylin-eosin(HE)staining was used to observe and analyze the pathological injury to lung tissue.The expres-sion of Lymphocyte antigen 6G(Ly6G)was detected by Immunofluorescence to reflect the infiltration of neutrophils.RESULTS Compared with the normal control group,LPS group and LPS+DXM group,IL-35 protein expression levels in lung homogenates of the other groups were more significant(P<0.01).Compared with the normal control group,lung coefficient,W/D ratio of lung tissue,LDH activity,mRNA levels and the protein levels of TNF-α,IL-6 and IL-1β in lung homogenates were significantly increased in the LPS group(P<0.01),accompanied by alveolar hemorrhage,alveolar wall thickening and neutro-phils infiltration.After IL-35 mRNA-LNP administration,lung coefficient,W/D ratio of lung tissue,LDH activity,mRNA levels and the protein levels of TNF-α,IL-6 and IL-1β in lung homogenates were signifi-cantly decreased(P<0.01),and alveolar hemorrhage,alveolar wall thickening and neutrophil infiltration were obviously improved.CONCLUSION IL-35 mRNA-LNP can express IL-35 protein in lung tissue of mice,and effectively improve LPS-induced ALI in mice by inhibiting the expression of proinflammatory factors.

Innate and adaptive immune responses initiated by infection depend on recognition and control of pathogens by macrophages,dendritic cells,T cells and so on.Notch signaling pathway is a highly conserved pathway,which is activated by interac-tion of receptors and ligands,thus coordinating important life processes of cells.At present,it has been confirmed that Notch signaling pathway is involved in development,differentiation,maturation and activation of many kinds of immune cells,and plays an important role in infectious diseases.In this review,we mainly focus on the role of Notch signaling pathway in immune regulation during different pathogens infection and its interaction with other signaling pathways.Additionally,therapeutic methods and challenges of developing Notch signaling as a target in infectious diseases are also discussed.

Objective:To overview the systematic reviews of prevention and management measures of medication errors, so as to provide evidence support for clinical decision-making for medical staff.Methods:Cochrane Library, Australia Joanna Briggs Institute Evidence-based Healthcare Center database, CINAHL, PubMed, Embase, CNKI, SinoMed, Wanfang database and VIP database were searched by computer to search for systematic reviews of prevention and management measures of medication errors, and the search period was from establishment of the databases to June 30, 2023. Two researchers with systematic evidence-based training applied A Measure Tool to Assess Systematic Reviews 2 (AMSTAR 2) to evaluate the literature quality, and Grades of Recommendations Assessment, Development and Evaluation (GRADE) was used to evaluate the quality of outcome indicators.Results:Finally, a total of 19 systematic reviews were included. The overall quality evaluation using AMSTAR 2 was relatively low, with one article rated as high-quality, one article rated as low-quality and 17 articles rated as extremely low-quality. According to the evidence quality evaluation results of GRADE system for 55 outcome indicators of 19 systematic reviews, 3 pieces of evidence were medium, 27 pieces of evidence were low and 25 pieces of evidence were extremely low, indicating an overall low quality of evidence.Conclusions:The related researches on prevention and management of medication errors have been carried out extensively, and the computer system is one of the effective measures to reduce medication errors. The effectiveness of measures such as administration process modification, doctor/nurse education and training, double check, pharmacist intervention, automated dispensing cabinet/ automated pump and drug display is still unclear and needs to be further confirmed by large sample size and high-quality studies.