BACKGROUND:In China,the patient population with osteonecrosis is large,and there is an urgent need to find new preventive targets to develop more effective treatment strategies.Metabolomics studies have shown that there is an association between human metabolites and osteonecrosis,but the causal relationship between blood metabolites and osteonecrosis has not yet been clarified.OBJECTIVE:To investigate the causal relationship between blood metabolites and osteonecrosis through two-sample Mendelian randomization analysis.METHODS:The public data of 486 blood metabolites(exposure factors)and osteonecrosis(outcome factors)were collected.Data of 486 blood metabolites were derived from a genome-wide association estimate for blood metabolites published in Nature Genetics in 2014,which covered 7 824 European adults.The single nucleotide polymorphism data for osteonecrosis were obtained from the FinnGen public database R11 dataset,containing information on a total of 431 614 samples and 21 306 430 single nucleotide polymorphism loci,with 1 788 cases of osteonecrosis and 429 826 controls,with all participants being of European descent.Mendelian randomization analysis(inverse variance weighting method,MR-Egger method,and weighted median method)was performed by Rstudio software,and then the heterogeneity test,horizontal pleiotropy test and Steiger directionality test were performed to ensure the robustness and reliability of the results.RESULTS AND CONCLUSION:(1)Sixteen blood metabolites were identified as having a significant causal relationship with osteonecrosis(Pinverse variance weighting＜Pfalse discovery rate＜0.05).(2)Eight blood metabolites increased the risk of osteonecrosis(including four known metabolites and four unknown metabolites),specifically pantothenate,beta-hydroxyisovalerate,hippurate,salicyluric glucuronide,X-08766,X-11452,X-12776 and X-14662.(3)Eight blood metabolites could reduce the risk of osteonecrosis(six known metabolites and two unknown metabolites),including cortisol,1-palmitoylglycerol(1-monopalmitin),pyroglutamyl glycine,2-stearoylglycerophosphocholine,p-cresol sulfate,ergothioneine,X-06307,X-12092.(4)The above results suggest that there is a causal relationship between 16 blood metabolites and osteonecrosis,which is expected to be a potential target for intervention in the occurrence and treatment of osteonecrosis in the future.(5)Despite the lack of relevant data from large-scale Asian populations at present,this study provides important reference value for the field of osteonecrosis in China based on European population data.In the future,domestic medical workers may be able to achieve precise intervention for osteonecrosis by regulating metabolite levels.In addition,based on the results of this study,relevant researchers can further explore the mechanism of action of metabolites in the treatment of osteonecrosis with traditional Chinese medicine,which not only helps to deepen the understanding of traditional Chinese medical therapies but also promotes the progress of integrated traditional Chinese and Western medicine research,driving the development of personalized treatment plans that are more suitable for the characteristics of the Chinese population.

BACKGROUND:In China,the patient population with osteonecrosis is large,and there is an urgent need to find new preventive targets to develop more effective treatment strategies.Metabolomics studies have shown that there is an association between human metabolites and osteonecrosis,but the causal relationship between blood metabolites and osteonecrosis has not yet been clarified.OBJECTIVE:To investigate the causal relationship between blood metabolites and osteonecrosis through two-sample Mendelian randomization analysis.METHODS:The public data of 486 blood metabolites(exposure factors)and osteonecrosis(outcome factors)were collected.Data of 486 blood metabolites were derived from a genome-wide association estimate for blood metabolites published in Nature Genetics in 2014,which covered 7 824 European adults.The single nucleotide polymorphism data for osteonecrosis were obtained from the FinnGen public database R11 dataset,containing information on a total of 431 614 samples and 21 306 430 single nucleotide polymorphism loci,with 1 788 cases of osteonecrosis and 429 826 controls,with all participants being of European descent.Mendelian randomization analysis(inverse variance weighting method,MR-Egger method,and weighted median method)was performed by Rstudio software,and then the heterogeneity test,horizontal pleiotropy test and Steiger directionality test were performed to ensure the robustness and reliability of the results.RESULTS AND CONCLUSION:(1)Sixteen blood metabolites were identified as having a significant causal relationship with osteonecrosis(Pinverse variance weighting＜Pfalse discovery rate＜0.05).(2)Eight blood metabolites increased the risk of osteonecrosis(including four known metabolites and four unknown metabolites),specifically pantothenate,beta-hydroxyisovalerate,hippurate,salicyluric glucuronide,X-08766,X-11452,X-12776 and X-14662.(3)Eight blood metabolites could reduce the risk of osteonecrosis(six known metabolites and two unknown metabolites),including cortisol,1-palmitoylglycerol(1-monopalmitin),pyroglutamyl glycine,2-stearoylglycerophosphocholine,p-cresol sulfate,ergothioneine,X-06307,X-12092.(4)The above results suggest that there is a causal relationship between 16 blood metabolites and osteonecrosis,which is expected to be a potential target for intervention in the occurrence and treatment of osteonecrosis in the future.(5)Despite the lack of relevant data from large-scale Asian populations at present,this study provides important reference value for the field of osteonecrosis in China based on European population data.In the future,domestic medical workers may be able to achieve precise intervention for osteonecrosis by regulating metabolite levels.In addition,based on the results of this study,relevant researchers can further explore the mechanism of action of metabolites in the treatment of osteonecrosis with traditional Chinese medicine,which not only helps to deepen the understanding of traditional Chinese medical therapies but also promotes the progress of integrated traditional Chinese and Western medicine research,driving the development of personalized treatment plans that are more suitable for the characteristics of the Chinese population.

BACKGROUND:Hormonal osteonecrosis of the femoral head is strongly associated with aging,but the regulatory targets and mechanisms are still unclear.Through bioinformatics combined with machine learning analysis and experimental verification,the key genes of hormonal osteonecrosis of the femoral head mediated by cell senescence will be identified,which will provide new ideas for the prevention and treatment of hormonal osteonecrosis of the femoral head.OBJECTIVE:To screen and validate the senescence core genes of hormonal osteonecrosis of the femoral head using bioinformatics analysis to explore its mechanism of action.METHODS:The GSE123568 dataset was obtained from the GPL15207 platform of the GEO database,which contained the gene expression profiles of peripheral serum samples of 30 hormonal osteonecrosis of the femoral head patients and 10 healthy controls.Data on 279 cellular senescence-related genes were obtained from the CellAge database.Differential analysis and weighted correlation network analysis(WGCNA)were performed on hormonal osteonecrosis of the femoral head gene profiles,and both were intersected with senescence-related genes and then concatenated to obtain hormonal osteonecrosis of the femoral head senescence potential genes,and GO and KEGG analyses were performed.The machine learning method screened out the pivotal genes,constructed nomogram model,and performed consensus clustering and immune infiltration analysis.Finally,clinical femoral samples were collected for validation by qPCR and western blot assay.RESULTS AND CONCLUSION:(1)41 potential genes were obtained,which were mainly enriched in biological processes such as aging and oxidative stress response,as well as FoxO and tumor necrosis factor signaling pathways.(2)The pivotal genes catalase,connective tissue growth factor,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were obtained after machine learning identification,and the predictive ability of nomogram model was good.(3)The patients were classified into three groups,namely a,b and c,by the consensus clustering analysis.Catalase,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were differentially expressed among the three molecular subtypes(P＜0.05).Results of immune infiltration showed that the abundance of immune cells,such as activated CD4+T cells,activated CD8+T cells,and eosinophils,differed among the three molecular subclasses(P＜0.05).(4)The results of qPCR and western blot assay showed that the expression of catalase,connective tissue growth factor,forkhead box protein O3,and mitogen-activated protein kinase kinase 11 was lower in hormonal osteonecrosis of the femoral head group compared to the control group(P＜0.05),and the expression of insulin receptor substrate 2 was elevated(P＜0.05).(5)It is concluded that through in-depth analysis combined with bioinformatics and machine learning,and further experimental verification,five hormonal osteonecrosis of the femoral head age-related hub genes were finally identified.These genes are catalase,connective tissue growth factor,forkhead box o3,insulin receptor substrate 2,and serine/threonine kinase 11.These genes may provide potential molecular targets for the prevention and treatment of hormonal osteonecrosis of the femoral head in the future by regulating the cellular aging process.

BACKGROUND:Hormonal osteonecrosis of the femoral head is strongly associated with aging,but the regulatory targets and mechanisms are still unclear.Through bioinformatics combined with machine learning analysis and experimental verification,the key genes of hormonal osteonecrosis of the femoral head mediated by cell senescence will be identified,which will provide new ideas for the prevention and treatment of hormonal osteonecrosis of the femoral head.OBJECTIVE:To screen and validate the senescence core genes of hormonal osteonecrosis of the femoral head using bioinformatics analysis to explore its mechanism of action.METHODS:The GSE123568 dataset was obtained from the GPL15207 platform of the GEO database,which contained the gene expression profiles of peripheral serum samples of 30 hormonal osteonecrosis of the femoral head patients and 10 healthy controls.Data on 279 cellular senescence-related genes were obtained from the CellAge database.Differential analysis and weighted correlation network analysis(WGCNA)were performed on hormonal osteonecrosis of the femoral head gene profiles,and both were intersected with senescence-related genes and then concatenated to obtain hormonal osteonecrosis of the femoral head senescence potential genes,and GO and KEGG analyses were performed.The machine learning method screened out the pivotal genes,constructed nomogram model,and performed consensus clustering and immune infiltration analysis.Finally,clinical femoral samples were collected for validation by qPCR and western blot assay.RESULTS AND CONCLUSION:(1)41 potential genes were obtained,which were mainly enriched in biological processes such as aging and oxidative stress response,as well as FoxO and tumor necrosis factor signaling pathways.(2)The pivotal genes catalase,connective tissue growth factor,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were obtained after machine learning identification,and the predictive ability of nomogram model was good.(3)The patients were classified into three groups,namely a,b and c,by the consensus clustering analysis.Catalase,forkhead box protein O3,insulin receptor substrate 2,and mitogen-activated protein kinase kinase 11 were differentially expressed among the three molecular subtypes(P＜0.05).Results of immune infiltration showed that the abundance of immune cells,such as activated CD4+T cells,activated CD8+T cells,and eosinophils,differed among the three molecular subclasses(P＜0.05).(4)The results of qPCR and western blot assay showed that the expression of catalase,connective tissue growth factor,forkhead box protein O3,and mitogen-activated protein kinase kinase 11 was lower in hormonal osteonecrosis of the femoral head group compared to the control group(P＜0.05),and the expression of insulin receptor substrate 2 was elevated(P＜0.05).(5)It is concluded that through in-depth analysis combined with bioinformatics and machine learning,and further experimental verification,five hormonal osteonecrosis of the femoral head age-related hub genes were finally identified.These genes are catalase,connective tissue growth factor,forkhead box o3,insulin receptor substrate 2,and serine/threonine kinase 11.These genes may provide potential molecular targets for the prevention and treatment of hormonal osteonecrosis of the femoral head in the future by regulating the cellular aging process.

BACKGROUND: Fibular fixation and tantalum rod implantation are two commonly used methods for the treatment of early osteonecrosis of the femoral head (ONFH), both of which can effectively delay or even reverse the progress of ONFH. However, further comparative evaluation on their mechanical properties and therapeutic efficacy is required.OBJECTIVE: To compare the clinical efficacy of fibular fixation and tantalum rod implantation on ONFH at early stage.METHODS: Fifty-eight patients (81 hips) suffered from ONFH with ARCO stage 1 and stage 2, and underwent fibular fixation (30 cases, 41 hips) or tantalum rod implantation (28 cases, 40 hips). Postoperatively, both groups were followed up for over 2 years. The Harris scores of the hip were compared between two groups before and after treatment. With femoral head collapse and the collapse distance > 4 mm as observation points, the survival rate of the femoral head was compared between two groups.RESULTS AND CONCLUSION: The postoperative Harris scores of the two groups were significantly improved than before (P < 0.05). With the appearance of femoral head collapse as the observation point, the Kaplan-Meier survival curve showed that the overall survival rate of the hip was 83% in the fibular fixation group and 65% in the tantalum rod implantation group. After examined by log-rank (Mantel-Cox), there was a significant difference in the survival rate of the hip at Stage IIC between two groups (P=0.0431). With > 4 mm collapse as the observation point, the Kaplan-Meier survival curve showed that overall survival rate of the hip was 95% in the fibular fixation group and 83% in the tantalum rod implantation group. After examined by log-rank (Mantel-Cox), there was a significant difference in the survival rate of the hip at Stage IIC between two groups (P=0.0418). To conclude, both fibular fixation and tantalum rods implantation applied to ONFH at early stage can effectively improve the hip function, and the survival rate of the hip at ARCO Stage IIC is better in patients undergoing fibular fixation than tantalum rod implantation.

BACKGROUND: Anterior and lateral columns of the femoral head integrity and stability have been reported to present a positive correlation with the prognosis of the osteonecrosis of the femoral head (ONFH) and efficacy of hip preserving. China-Japan Friendship Hospital Classification stressed that the retention of the anterior and lateral columns of the femoral head was of great importance to avoid the femoral head collapse, but there is little reported on its three-dimensional (3D) digital model.OBJECTIVE: To explore the method of establishing highly-simulated 3D model of China-Japan Friendship Hospital Classification for ONFH.METHODS: 3D reconstruction of normal and necrotic femoral head was established according to the CT and MRI of individuals with normal femoral head. A healthy adult male volunteer was selected and his CT image was collected; three adult male patients with types L1, L2 and L3 ONFH were selected, whose MRI images were obtained, respectively. 3D solid models were established based on CT and MRI data on Mimics 15.0, Geomagic Studio 13, Geomagic Design X, Solidworks2014 and Abauqus6.14 software. RESULTS AND CONCLUSION: A highly-simulated 3D model of L type ONFH was established, including the corticaland cancellous bones of ilium and proximal femur, articular cartilage, necrotic articular cartilage, muscles, joint capsuleand ligaments. It clearly showed the spatial structures of L type ONFH, by which different surgical programs could be compared in a virtual environment for selecting an appropriate treatment strategy to improve the success rate of hip preserving. The highly-simulated 3D model paves ways for surgical simulation and biomechanical analysis.

Objective To investigate the effects of injection of citicoline into Zusanli (ST36) acupoint on neural function and expression of growth associated protein-43 (GAP-43) in rats with traumatic brain injury. Methods 40 healthy adult male Sprague-Dawley rats were randomly divided into 5 groups: sham-operated group (group A, n=8), acupoint injection of citicoline group (group B, n=8), acupoint injection of saline group (group C, n=8), intraperitoneal injection of citicoline group (group D, n=8) and intraperitoneal injection of saline group (group E, n=8). Opened brain trauma was induced with the modified Feeney method in the groups B, C, D and E, and were treated as design, once a day for 14 days. They were assessed with nervous function score and open-field test before and 8, 14, 15, and 22 days after injury. The expression of GAP-43 in the brain were detected with immunohistochemistry 28 days after injury. Results The nervous function scores and open-field test scores improved more significantly in the group B than in the groups C, D and E (P<0.05). The expression of GAP-43 increased in the group B than in the groups C, D and E (P<0.05). Conclusion Acupoint injection of citicoline into Zusanli may improve the expressions of GAP-43 to promote the recovery of neural function in rats after traumatic brain injury.

BACKGROUND:Bone marrow mesenchymal stem cells have the potential of self-proliferation and multi-directional differentiation, while mesenchymal stem cells are few in adult bone marrow. In vitro purification, amplification and osteoinduction are very important for the research of bone tissue engineering. OBJECTIVE:To establish a simple and reliable in vitro cultivation and identification system of adult bone marrow mesenchymal stem cells, and to induce the mesenchymal stem cells to differentiate into osteoblasts. METHODS:Bone marrow were extracted from adult anterior superior iliac, the density gradient centrifugation and adhesion method were used to isolate, purify, culture and amplify the bone marrow mesenchymal stem cells. Osteogenic medium was prepared by mixing appropriate amount of dexamethasone,β-glycerophosphate and ascorbic acid C. The cells were divided into osteoinduction group and blank control group for observation.
RESULTS AND CONCLUSION:Adult bone marrow mesenchymal stem cells were in typical long spindle-shape. The cells grew into rapid proliferation phase at 8-11 days and the growth curve was S-shape. CD44 and CD90 were in positive expression, while CD34 and CD45 were negative. The alkaline phosphatase activity was increased with culturing time prolonging, and reached the summit at the 12th day. The alkaline phosphatase activities of osteoinduction group were higher than those in the blank control group at different time points. These results suggested that in vitro cultivation, identification and osteoinduction system could obtain mesenchymal stem cells with high purity and good osteogenic differentiation capacity.

Objective To explore the effect of early Chinese medicine intervention on motor function and balance capability of patients with traumatic brain injury (TBI). Methods 55 cases of hospitalized TBI patients were randomly divided into observation group (n=27) and control group (n=28). The control group accepted routine rehabilitation program, while the observation group accepted Chinese medicine therapy in addition for 8 weeks. The motor function and balance capability were tested with Fugl-Meyer Assessment (FMA) and Berg Balance Scale (BBS) respectively before and after treatment. Results The scores of FMA and BBS significantly increased after treatment (P<0.01), and they were higher in the observation group than in the control group (P<0.05). Conclusion Chinese medicine combined with routine rehabilitation program is effective on motor dysfunction after TBI.

Objective To observe the effects of Chinese Traditional Medicine on cognitive functions in the early stage of traumatic braininjury (TBI). Methods 49 inpatients with TBI were divided into treatment group (n=25) and control group (n=24) randomly. The controlgroup accepted routine rehabilitation, while the treatment group accepted Chinese Traditional Medicine in addition. They were assessed withLoewenstein Occupational Therapy Cognitive Assessment (LOTCA) before and after treatment. Results All the scores of LOTCA of treatmentgroup significantly improved after treatment (P<0.01), as well as in control group except "Categories". Most scores of LOTCA improvedmore in the treatment group than in the control group (P<0.05). Conclusion Chinese Traditional Medicine can improve the cognitivefunction after TBI.