BACKGROUND:Relatively or absolutely active bone resorption function of osteoclasts is one of the causative factors of osteoporosis. Therefore,how to inhibit the formation of osteoclasts and reduce the bone resorption activity is a key element in the prevention and treatment of osteoporosis. Liquiritin,which is derived from licorice,plays a role in the clinical treatment of bone diseases,but there are fewer studies addressing the application of liquiritin in osteoporosis and the mechanism is unknown.OBJECTIVE:To confirm,through both in vivo and in vitro experiments,that liquiritin inhibits osteoclast differentiation and alleviates bone loss.METHODS:Cell counting kit-8 was used to detect whether Liquiritin exerts toxic or proliferative effects on mouse bone marrow-derived macrophages,and tartrate-resistant acid phosphatase staining was performed to observe the effect of liquiritin in inhibiting osteoclast differentiation. The affinity of liquiritin binding to proteins related to osteoclast differentiation was verified by network pharmacology. RT-PCR and western blot assays were performed to detect the inhibitory effects of liquiritin on osteoclast-specific protein and gene expression as well as relevant signaling pathways. Finally,the mitigating effect of liquiritin on bone loss was verified in the C57BL/6J mouse osteoporosis model.RESULTS AND CONCLUSION:Liquiritin,at concentrations of 20 μmol/L and below,could inhibit the formation and differentiation of osteoclasts. Concurrently,it exhibited a high affinity with osteoclast-specific proteins such as nuclear factor of activated T-cells 1,Cathepsin K,c-Fos,and matrix metalloproteinase 9,and reduced the relative expression levels of these genes and proteins. Liquiritin could also effectively lower the phosphorylation expression level of JNK in the MAPK signaling pathway at the 15th,30th,45th,and 60th minutes,and it could salvage the degradation of nuclear factor-κB inhibitor α in the nuclear factor-κB signaling pathway at the 60th minute. In vivo experiments demonstrated that liquiritin could mitigate bone loss caused by osteoclasts and improve parameters related to trabecular bone. To conclude,liquiritin possesses the capacity to inhibit osteoclast differentiation and alleviate bone loss,thereby exerting a protective role against osteoporosis.

Objective::There is limited evidence regarding the choice of P2Y 12 receptor inhibitors as a component of dual antiplatelet therapy in patients with left main (LM) disease undergoing percutaneous coronary intervention (PCI). This study aimed to evaluate long-term clinical outcomes of ticagrelor- vs. clopidogrel-based dual antiplatelet therapy strategy in acute coronary syndrome (ACS) patients undergoing LM PCI. Methods::This is a post-hoc analysis from a prospective, single-center, real-world PCI registry. A total of 1,163 patients discharged post-ACS who underwent LM PCI and received ticagrelor or clopidogrel between March 2016 and March 2019 were included in the study. The primary endpoint was ischemic events at 12 months, including cardiac death, myocardial infarction, or stroke. Secondary outcomes included all-cause death and Bleeding Academic Research Consortium types 2, 3, and 5, and types 3 and 5 bleeding. Propensity score matching was used to adjust for bias due to confounders between the 2 groups.Results::The ticagrelor and clopidogrel groups comprised 529 (45.49%) and 634 (54.51%) patients, respectively. During the follow-up period, the rate of ischemic events was significantly lower with ticagrelor than with clopidogrel before (1.32% (7/529) vs. 3.63% (23/634), P = 0.013,6) and after propensity score matching (1.41% (6/425) vs. 4.00% (17/425), P = 0.020,1). The rates of all-cause death, Bleeding Academic Research Consortium-defined type 2, 3, and 5 bleeding, and type 3 and 5 bleeding were similar between the ticagrelor group and clopidogrel group before or after propensity score matching adjustment (all P > 0.05). Conclusion::Among patients with ACS undergoing LM PCI, ticagrelor use was associated with ischemic events benefit without excessive risk of bleeding at 12 months compared with clopidogrel.

Objective To investigate the role of luteolin(Lut)in regulating reactive oxygen species(ROS)levels through nuclear factor erythroid 2-related factor 2(NFE2L2)/cystine glutamate antitransporter(x-CT)/glutathione peroxidase 4(GPX4)signaling axis to inhibit the viability of glioblastoma and promote apoptosis.Methods U87 MG and U251 cells were cultured in vitro.The CCK-8 assay was used to detect cell survival rates after 48 hours of treatment with different concentrations(0,6.25,12.5,25,50 and 100 μmol/L)of Lut.According to whether cells were treated with Lut,cells were divided into the U87 control group,the U87 Lut group,the U251 control group and the U251 Lut group.The half-maximal inhibitory concentration(IC50)at 48 hours was used as the unified treatment concentration for subsequent experiments.The apoptosis level of cells was detected by flow cytometry double staining method.Changes of reactive oxygen species(ROS)levels in cells were detected by the DCFH-DA method.Molecular docking was conducted using AutoDock software to verify the proteins related to the Lut and oxidative stress pathway.Real-time fluorescence quantitative reverse transcription(RT-qPCR)was used to detect the mRNA levels of NFE2L2 and GPX4.The expression levels of NFE2L2,x-CT and GPX4 proteins were detected by Western blot assay.Results After U87 MG and U251 cells were treated with Lut for 48 hours,the cell viability was significantly inhibited,and with the increase of Lut concentration,the cell viability decreased(P＜0.05).Compared with the U87 control group and the U251 control group respectively,the apoptosis rate of cells increased in the U87 Lut group and the U251 Lut group,the green fluorescence intensity was enhanced,and the intracellular ROS level was upregulated(P＜0.05).Results of molecular docking showed that Lut was tightly bound to NFE2L2,x-CT and GPX4.The results of RT-qPCR and Western blot assay showed that compared with the U87 control group and the U251 control group respectively,the protein and mRNA levels of NFE2L2 and GPX4 in cells of the U87 Lut group and the U251 Lut group,as well as the expression level of x-CT protein,decreased(P＜0.05).Conclusion Lut regulates ROS levels through the NFE2L2/x-CT/GPX4 signaling axis to inhibit the viability of glioblastoma and promote cell apoptosis.

Objective To investigate the changes and clinical significance of synovial fluid Asporin level in patients with tem-poromandibular joint disorders(TMD).Methods A total of 48 TMD patients who were treated in our hospital from January 2021 to December 2023 due to irritant pain and mouth opening limitation were randomly selected as the observation group,and 48 healthy vol-unteers were selected as the control group.The synovial fluid Asporin levels of the two groups were detected by enzyme-linked immu-nosorbent assay(ELISA).The difference of synovial fluid Asporin levels between the two groups was compared.The correlation be-tween the synovial fluid Asporin levels and the clinical symptoms of TMD was analyzed.Results The synovial fluid Asporin level in the experimental group was significantly higher than that in the control group(P＜0.05).The synovial fluid Asporin level was positively correlated with the pain degree of TMD patients(Rs=0.825,P＜0.001),negatively correlated with the degree of mouth opening(Rs=-0.945,P＜0.001).Conclusion The level of Asporin in synovial fluid of TMD patients was significantly increased.The level of As-porin in synovial fluid of TMD patients is correlated with the clinical symptoms of TMD,which provides a basis for the diagnosis and evaluation of TMD.

Objective::There is limited evidence regarding the choice of P2Y 12 receptor inhibitors as a component of dual antiplatelet therapy in patients with left main (LM) disease undergoing percutaneous coronary intervention (PCI). This study aimed to evaluate long-term clinical outcomes of ticagrelor- vs. clopidogrel-based dual antiplatelet therapy strategy in acute coronary syndrome (ACS) patients undergoing LM PCI. Methods::This is a post-hoc analysis from a prospective, single-center, real-world PCI registry. A total of 1,163 patients discharged post-ACS who underwent LM PCI and received ticagrelor or clopidogrel between March 2016 and March 2019 were included in the study. The primary endpoint was ischemic events at 12 months, including cardiac death, myocardial infarction, or stroke. Secondary outcomes included all-cause death and Bleeding Academic Research Consortium types 2, 3, and 5, and types 3 and 5 bleeding. Propensity score matching was used to adjust for bias due to confounders between the 2 groups.Results::The ticagrelor and clopidogrel groups comprised 529 (45.49%) and 634 (54.51%) patients, respectively. During the follow-up period, the rate of ischemic events was significantly lower with ticagrelor than with clopidogrel before (1.32% (7/529) vs. 3.63% (23/634), P = 0.013,6) and after propensity score matching (1.41% (6/425) vs. 4.00% (17/425), P = 0.020,1). The rates of all-cause death, Bleeding Academic Research Consortium-defined type 2, 3, and 5 bleeding, and type 3 and 5 bleeding were similar between the ticagrelor group and clopidogrel group before or after propensity score matching adjustment (all P > 0.05). Conclusion::Among patients with ACS undergoing LM PCI, ticagrelor use was associated with ischemic events benefit without excessive risk of bleeding at 12 months compared with clopidogrel.

Objective To investigate the role of luteolin(Lut)in regulating reactive oxygen species(ROS)levels through nuclear factor erythroid 2-related factor 2(NFE2L2)/cystine glutamate antitransporter(x-CT)/glutathione peroxidase 4(GPX4)signaling axis to inhibit the viability of glioblastoma and promote apoptosis.Methods U87 MG and U251 cells were cultured in vitro.The CCK-8 assay was used to detect cell survival rates after 48 hours of treatment with different concentrations(0,6.25,12.5,25,50 and 100 μmol/L)of Lut.According to whether cells were treated with Lut,cells were divided into the U87 control group,the U87 Lut group,the U251 control group and the U251 Lut group.The half-maximal inhibitory concentration(IC50)at 48 hours was used as the unified treatment concentration for subsequent experiments.The apoptosis level of cells was detected by flow cytometry double staining method.Changes of reactive oxygen species(ROS)levels in cells were detected by the DCFH-DA method.Molecular docking was conducted using AutoDock software to verify the proteins related to the Lut and oxidative stress pathway.Real-time fluorescence quantitative reverse transcription(RT-qPCR)was used to detect the mRNA levels of NFE2L2 and GPX4.The expression levels of NFE2L2,x-CT and GPX4 proteins were detected by Western blot assay.Results After U87 MG and U251 cells were treated with Lut for 48 hours,the cell viability was significantly inhibited,and with the increase of Lut concentration,the cell viability decreased(P＜0.05).Compared with the U87 control group and the U251 control group respectively,the apoptosis rate of cells increased in the U87 Lut group and the U251 Lut group,the green fluorescence intensity was enhanced,and the intracellular ROS level was upregulated(P＜0.05).Results of molecular docking showed that Lut was tightly bound to NFE2L2,x-CT and GPX4.The results of RT-qPCR and Western blot assay showed that compared with the U87 control group and the U251 control group respectively,the protein and mRNA levels of NFE2L2 and GPX4 in cells of the U87 Lut group and the U251 Lut group,as well as the expression level of x-CT protein,decreased(P＜0.05).Conclusion Lut regulates ROS levels through the NFE2L2/x-CT/GPX4 signaling axis to inhibit the viability of glioblastoma and promote cell apoptosis.

Objective To investigate the changes and clinical significance of synovial fluid Asporin level in patients with tem-poromandibular joint disorders(TMD).Methods A total of 48 TMD patients who were treated in our hospital from January 2021 to December 2023 due to irritant pain and mouth opening limitation were randomly selected as the observation group,and 48 healthy vol-unteers were selected as the control group.The synovial fluid Asporin levels of the two groups were detected by enzyme-linked immu-nosorbent assay(ELISA).The difference of synovial fluid Asporin levels between the two groups was compared.The correlation be-tween the synovial fluid Asporin levels and the clinical symptoms of TMD was analyzed.Results The synovial fluid Asporin level in the experimental group was significantly higher than that in the control group(P＜0.05).The synovial fluid Asporin level was positively correlated with the pain degree of TMD patients(Rs=0.825,P＜0.001),negatively correlated with the degree of mouth opening(Rs=-0.945,P＜0.001).Conclusion The level of Asporin in synovial fluid of TMD patients was significantly increased.The level of As-porin in synovial fluid of TMD patients is correlated with the clinical symptoms of TMD,which provides a basis for the diagnosis and evaluation of TMD.

Objective:To investigate the promoting effect of individualized instruction combined with a blended learning model (IIBLM) in continuing training of neurology.Methods:A total of 93 trainees who received continuing training in Department of Neurology, The First Affiliated Hospital of Sun Yat-sen University, from August 2022 to December 2024 were enrolled as subjects. The 50 trainees registered since January 2024 were enrolled as observation group and received IIBLM teaching, including sub-specialty modular training, a cycle-adaptive cultivation system, a "mutual-selection" mentorship program, an on/off-line dual-track curriculum system, a dynamic course allocation mechanism based on "mutual selection", and a competency growth evaluation system, while the 43 trainees registered before January 2024 were enrolled as control group and received traditional teaching. A questionnaire survey and comprehensive competency assessments were performed to evaluate the effect of teaching, and the t-test, the chi-square test, and the qualitative analysis were used for statistical analysis. Results:Compared with the control group, the experimental group showed systematic improvements in clinical contents, theoretical curriculum, faculty competency, and workflow arrangement during continuing training, with a significant difference in the score of workflow arrangement between the two groups [(9.58±0.67) vs. (9.07±1.44), t=-2.13, P=0.037]. The observation group had a score of (97.70±1.30) for dynamic course allocation, an overall satisfaction rate of 97.15%, and a course benefit rate of 97.55%. The qualitative analysis showed that the trainees in the control group mainly complained of course monotony, while those in the observation group expected to enhance interdisciplinary integration and the cultivation of scientific research abilities. In addition, the results of competency assessment showed a continuous improvement in teaching effect after reform, with no significant difference. Conclusions:IIBLM teaching effectively enhances professional qualities, clinical competency, and the degree of satisfaction with courses among the trainees receiving continuing training, and it also revealed the necessity of interdisciplinary collaborative teaching and the integration of research and clinical practice.

ObjectiveTo investigate the mechanism of modified Si Junzitang and Shashen Maidong Tang ［Yiqi Yangyin Jiedu prescription （YQYYJD）］ in enhancing the sensitivity of cisplatin in epidermal growth factor receptor tyrosine kinase inhibitor （EGFR-TKI）-resistant lung adenocarcinoma cells based on aerobic glycolysis. MethodsThe effects of different concentrations of YQYYJD （0， 2， 3， 4， 5， 6， 7， 8 g·L^-1） and cisplatin （0， 3， 6， 9， 12， 15， 18， 21， 24， 27 mg·L^-1） on the proliferation and activity of PC9/GR cells were detected by the cell counting kit-8 （CCK-8） assay after 24 hours of intervention. The half-maximal inhibitory concentration （IC₅₀） for PC9/GR cells was calculated to determine the concentrations used in subsequent experiments. PC9/GR cells were divided into blank group （complete medium）， YQYYJD group （5 g·L^-1）， cisplatin group （12 mg·L^-1）， and combined group （YQYYJD 5 g·L^-1 + cisplatin 12 mg·L^-1）. After 24 hours of intervention， cell viability was measured using CCK-8 assay. Cell proliferation was assessed by colony formation assay， and cell migration was evaluated by scratch and Transwell assays. Glucose consumption， lactate production， and adenosine triphosphate （ATP） levels were measured by colorimetric assays. The expression levels of glycolysis-related proteins， including hexokinase 2 （HK2）， phosphofructokinase P （PFKP）， pyruvate kinase M2 （PKM2）， lactate dehydrogenase A （LDHA）， glucose transporter 1 （GLUT1）， and monocarboxylate transporter 4 （MCT4）， were determined by Western blot. ResultsBoth YQYYJD and cisplatin inhibited the viability of PC9/GR cells in a concentration-dependent manner. The IC₅₀ of PC9/GR cells for YQYYJD and cisplatin were 5.15 g·L^-1 and 12.91 mg·L^-1， respectively. In terms of cell proliferation， compared with the blank group， the cell survival rate and the number of colonies formed in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in cell survival rate and colony formation （P<0.01）. In terms of cell migration， compared with the blank group， the cell migration rate and the number of cells passing through the Transwell membrane in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group exhibited a further significant reduction in cell migration rate and the number of cells passing through the Transwell membrane （P<0.01）. In terms of glycolysis， compared with the blank group， glucose consumption， lactate production， and ATP levels in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in glucose consumption， lactate production， and ATP levels （P<0.05）. Compared with the blank group， the protein expression levels of HK2， PFKP， PKM2， and LDHA in the YQYYJD， cisplatin， and combined groups were significantly decreased （P<0.01）. The combined group showed a further significant reduction in the expression levels of these proteins compared with the YQYYJD and cisplatin groups （P<0.01）. No significant differences were observed in the protein expression levels of GLUT1 and MCT4 among the groups. ConclusionYQYYJD can synergistically inhibit the proliferation and migration of PC9/GR cells and enhance their sensitivity to cisplatin. The mechanism may be related to the downregulation of the expression of glycolysis-related rate-limiting enzymes， including HK2， PFKP， PKM2， and LDHA， thereby inhibiting glycolysis.

BACKGROUND:Relatively or absolutely active bone resorption function of osteoclasts is one of the causative factors of osteoporosis. Therefore,how to inhibit the formation of osteoclasts and reduce the bone resorption activity is a key element in the prevention and treatment of osteoporosis. Liquiritin,which is derived from licorice,plays a role in the clinical treatment of bone diseases,but there are fewer studies addressing the application of liquiritin in osteoporosis and the mechanism is unknown.OBJECTIVE:To confirm,through both in vivo and in vitro experiments,that liquiritin inhibits osteoclast differentiation and alleviates bone loss.METHODS:Cell counting kit-8 was used to detect whether Liquiritin exerts toxic or proliferative effects on mouse bone marrow-derived macrophages,and tartrate-resistant acid phosphatase staining was performed to observe the effect of liquiritin in inhibiting osteoclast differentiation. The affinity of liquiritin binding to proteins related to osteoclast differentiation was verified by network pharmacology. RT-PCR and western blot assays were performed to detect the inhibitory effects of liquiritin on osteoclast-specific protein and gene expression as well as relevant signaling pathways. Finally,the mitigating effect of liquiritin on bone loss was verified in the C57BL/6J mouse osteoporosis model.RESULTS AND CONCLUSION:Liquiritin,at concentrations of 20 μmol/L and below,could inhibit the formation and differentiation of osteoclasts. Concurrently,it exhibited a high affinity with osteoclast-specific proteins such as nuclear factor of activated T-cells 1,Cathepsin K,c-Fos,and matrix metalloproteinase 9,and reduced the relative expression levels of these genes and proteins. Liquiritin could also effectively lower the phosphorylation expression level of JNK in the MAPK signaling pathway at the 15th,30th,45th,and 60th minutes,and it could salvage the degradation of nuclear factor-κB inhibitor α in the nuclear factor-κB signaling pathway at the 60th minute. In vivo experiments demonstrated that liquiritin could mitigate bone loss caused by osteoclasts and improve parameters related to trabecular bone. To conclude,liquiritin possesses the capacity to inhibit osteoclast differentiation and alleviate bone loss,thereby exerting a protective role against osteoporosis.