Objective To explore the differential expression and mechanisms of bone formation-related genes in osteoporosis(OP)leveraging bioinformatics and machine learning methodologies;and to predict the active ingredients of ...Objective To explore the differential expression and mechanisms of bone formation-related genes in osteoporosis(OP)leveraging bioinformatics and machine learning methodologies;and to predict the active ingredients of targeted traditional Chinese medicine(TCM)herbs.Methods The Gene Expression Omnibus(GEO)and GeneCards databases were employed to conduct a comprehensive screening of genes and disease-associated loci pertinent to the pathogenesis of OP.The R package was utilized as the analytical tool for the identification of differentially expressed genes.Least absolute shrinkage and selection operator(LASSO)logis-tic regression analysis and support vector machine-recursive feature elimination(SVM-RFE)algorithm were employed in defining the genetic signature specific to OP.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses for the selected pivotal genes were conducted.The cell-type identification by estimating rela-tive subsets of RNA transcripts(CIBERSORT)algorithm was leveraged to examine the infiltra-tion patterns of immune cells;with Spearman’s rank correlation analysis utilized to assess the relationship between the expression levels of the genes and the presence of immune cells.Coremine Medical Database was used to screen out potential TCM herbs for the treatment of OP.Comparative Toxicogenomics Database(CTD)was employed for forecasting the TCM ac-tive ingredients targeting the key genes.AutoDock Vina 1.2.2 and GROMACS 2020 softwares were employed to conclude analysis results;facilitating the exploration of binding affinity and conformational dynamics between the TCM active ingredients and their biological targets.Results Ten genes were identified by intersecting the results from the GEO and GeneCards databases.Through the application of LASSO regression and SVM-RFE algorithm;four piv-otal genes were selected:coat protein(CP);kallikrein 3(KLK3);polymeraseγ(POLG);and transient receptor potential vanilloid 4(TRPV4).GO and KEGG pathway enrichment analy-ses revealed that these trait genes were predominantly engaged in the regulation of defense response activation;maintenance of cellular metal ion balance;and the production of chemokine ligand 5.These genes were notably associated with signaling pathways such as ferroptosis;porphyrin metabolism;and base excision repair.Immune infiltration analysis showed that key genes were highly correlated with immune cells.Macrophage M0;M1;M2;and resting dendritic cell were significantly different between groups;and there were signifi-cant differences between different groups(P<0.05).The interaction counts of resveratrol;curcumin;and quercetin with KLK3 were 7;3;and 2;respectively.It shows that the interac-tions of resveratrol;curcumin;and quercetin with KLK3 were substantial.Molecular docking and molecular dynamics simulations further confirmed the robust binding affinity of these bioactive compounds to the target genes.Conclusion Pivotal genes including CP;KLK3;POLG;and TRPV4;exhibited commendable significant prognostic value;and played a crucial role in the diagnostic assessment of OP.Resveratrol;curcumin;and quercetin;natural compounds found in TCM;showed promise in their potential to effectively modulate the bone-forming gene KLK3.This study provides a sci-entific basis for the interpretation of the pathogenesis of OP and the development of clinical drugs.展开更多
[Objective] This study aimed to investigate the differences in morphological structure of tibias between sick and normal ostrich chicks. [Method] Normal and sick ostrich chicks aged 90 days were used in this research ...[Objective] This study aimed to investigate the differences in morphological structure of tibias between sick and normal ostrich chicks. [Method] Normal and sick ostrich chicks aged 90 days were used in this research for bone mineral density determination, bone demineralization paraffin section preparation （HE staining）, bone abrasive disc preparation and scanning electron microscopy （SEM） observation, to analyze the differences in microstructure and ultrastructure of tibias between sick ostrich chicks and normal ones. [Result] Bone mineral density of ostrich chicks suffering from leg disease was much lower than that of the normal ones. Compared with normal tibias, less trabeculae were observed in sick tibias, which were attenuated and began to disappear. In addition, trabeculae micro-fracture could be observed under scanning electron microscope; resorption pits on bone slices formed by osteoclast could be observed under scanning electronic microscope. [Conclusion] These results indicate that the sick tibias all have lower bone density and trabeculae microfracture phenomenon in tibias with different diseases, which might be resulted from osteoporosis.展开更多
In order to analyze skull variation in the genus Macaca ,seventy seven craniodental variables were taken from eleven species.They were first defined seven functional units comprising three anatomical regions.Twenty s...In order to analyze skull variation in the genus Macaca ,seventy seven craniodental variables were taken from eleven species.They were first defined seven functional units comprising three anatomical regions.Twenty seven variables were finally selected to carry out the morphology of the whole skull.The data,organized in these ways,were examined to discover variations between and within the various species.The methods used were Principal Components Analysis (PCA) and Discriminant Function Analysis (DFA).PCAs of the functional units anatomical regions,and the whole skull provided similar,though not identical,separations of species clusters in both sexes separately.These differences in structure could be related to size,sexual dimorphism,diet,ecology,classification and phylogeny.The question of size should have been easy to settle.Unfortunately,this is not the case.In this study where the raw data are measurements of the specimens,the main differences should be size.However,the size differences seem to occur in both the first and second (independent) multivariate axes.In some analyses the size differences between the species are biggest and appear in the first axis.In other analyses it is the separation between the sexes (and these too are largely size) that are the biggest and appear in the first axis.Yet in other analyses,both of these size separations,though still orthogonal to one another,present in the combination of the first two axes.This certainly implies that a single axis of body size is not present and that shape differences have not been isolated form size differences.It also implies that sexual dimorphism is a complex matter.As a result,the question of the relationships between the species is therefore also complex.One cluster of species that includes M fascicularis,M sinica and M radiata was significantly isolated from all others regardless of level of analysis.This relationship is quite different from that proposed on the anatomy of the reproductive organs (Delson,1980;Fooden,1976,1980).展开更多
Objective To Investigate stress adaptability of freeze-dried bone allograft.Methods Cortical and cancellous allograft were transplanted to each side of the midshaft diaphyseal ulna in two groups of 28 animals.The left...Objective To Investigate stress adaptability of freeze-dried bone allograft.Methods Cortical and cancellous allograft were transplanted to each side of the midshaft diaphyseal ulna in two groups of 28 animals.The left transplanted allograft was free from fixation and bore a normal physiological lcad,while the right transplanted allograft was protected from loading by a simple external fixator and bore less load.Animals were sacrificed at the 2nd,4th,8th,16th week after transplantation and specimens were taken out for bone histomorphometry studies and analysis of collagen gene expression by in situ Cdna-Mrna hybridization.Results Labeled surface(LS)and bone mineral apposition rate(MAR)of the normally loaded graft-host bone interface were significantly higher than that of the less loaded side at the 4th,8th,16th week after transplantation.Parameters reflecting the internal repair process of the allograft,such as LS in cortical and cancellous bone or MAR in cortical bone of the normally loaded side were significantly higher than those of the less loaded side at the 16th week after transplantation.The result of in situ hybridization indicated that more osteoblast-like cells expressing the type Ⅰ collagen gene were found in the interface or interior of normally loaded grafts.Conclusion The stimulus of physiologic load can accelerate the early union of allograft-host bone interface and later new bone creep substitution to the necrotic allograft.展开更多
Objective: To evaluate the osteogenic potential of bone morphogenetic protein (BMP)-2 gene transfected goat bone marrow-derived mesenchymal stem cells (MSCs). Methods: Goat bone marrow- derived MSCs were transfected b...Objective: To evaluate the osteogenic potential of bone morphogenetic protein (BMP)-2 gene transfected goat bone marrow-derived mesenchymal stem cells (MSCs). Methods: Goat bone marrow- derived MSCs were transfected by Adv-human bone morphogenetic protein (hBMP)-2 gene(Group 1), Adv-beta gal transfected MSCs (Group 2)and uninfected MSCs(Group 3). Western blot analysis, alkaline phosphatase staining, Von Kossa staining and transmission electron microscopy were adopted to determine the phenotype of MSCs. Then the cells were injected into thigh muscles of the nude mice. Radiographical and histological evaluations were performed at different intervals. Results: Only Adv-hBMP-2 transfected MSCs produced hBMP-2. These cells were positive for alkaline phosphatase staining at the 12th day and were positive for Von Kossa staining at the 16th day after gene transfer. Electron microscopic observation showed that there were more rough endoplasmic reticulum, mitochondria and lysosomes in Adv-hBMP-2 transfected MSCs compared to MSCs of other two groups. At the 3rd and 6th weeks after cell injection, ectopic bones were observed in muscles of nude mice of Group 1. Only fibrous tissue or a little bone was found in other two groups. Conclusions: BMP-2 gene transfected MSCs can differentiate into osteoblasts in vitro and induce bone formation in vivo.展开更多
基金National Natural Science Foundation of China(81960877).
文摘Objective To explore the differential expression and mechanisms of bone formation-related genes in osteoporosis(OP)leveraging bioinformatics and machine learning methodologies;and to predict the active ingredients of targeted traditional Chinese medicine(TCM)herbs.Methods The Gene Expression Omnibus(GEO)and GeneCards databases were employed to conduct a comprehensive screening of genes and disease-associated loci pertinent to the pathogenesis of OP.The R package was utilized as the analytical tool for the identification of differentially expressed genes.Least absolute shrinkage and selection operator(LASSO)logis-tic regression analysis and support vector machine-recursive feature elimination(SVM-RFE)algorithm were employed in defining the genetic signature specific to OP.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses for the selected pivotal genes were conducted.The cell-type identification by estimating rela-tive subsets of RNA transcripts(CIBERSORT)algorithm was leveraged to examine the infiltra-tion patterns of immune cells;with Spearman’s rank correlation analysis utilized to assess the relationship between the expression levels of the genes and the presence of immune cells.Coremine Medical Database was used to screen out potential TCM herbs for the treatment of OP.Comparative Toxicogenomics Database(CTD)was employed for forecasting the TCM ac-tive ingredients targeting the key genes.AutoDock Vina 1.2.2 and GROMACS 2020 softwares were employed to conclude analysis results;facilitating the exploration of binding affinity and conformational dynamics between the TCM active ingredients and their biological targets.Results Ten genes were identified by intersecting the results from the GEO and GeneCards databases.Through the application of LASSO regression and SVM-RFE algorithm;four piv-otal genes were selected:coat protein(CP);kallikrein 3(KLK3);polymeraseγ(POLG);and transient receptor potential vanilloid 4(TRPV4).GO and KEGG pathway enrichment analy-ses revealed that these trait genes were predominantly engaged in the regulation of defense response activation;maintenance of cellular metal ion balance;and the production of chemokine ligand 5.These genes were notably associated with signaling pathways such as ferroptosis;porphyrin metabolism;and base excision repair.Immune infiltration analysis showed that key genes were highly correlated with immune cells.Macrophage M0;M1;M2;and resting dendritic cell were significantly different between groups;and there were signifi-cant differences between different groups(P<0.05).The interaction counts of resveratrol;curcumin;and quercetin with KLK3 were 7;3;and 2;respectively.It shows that the interac-tions of resveratrol;curcumin;and quercetin with KLK3 were substantial.Molecular docking and molecular dynamics simulations further confirmed the robust binding affinity of these bioactive compounds to the target genes.Conclusion Pivotal genes including CP;KLK3;POLG;and TRPV4;exhibited commendable significant prognostic value;and played a crucial role in the diagnostic assessment of OP.Resveratrol;curcumin;and quercetin;natural compounds found in TCM;showed promise in their potential to effectively modulate the bone-forming gene KLK3.This study provides a sci-entific basis for the interpretation of the pathogenesis of OP and the development of clinical drugs.
基金Supported by Specialized Research Fund for The Doctoral Program of Huazhong Agricultural University(200805040023)Natural Science Foundation of China(NSFC)(30471249,30972152)~~
文摘[Objective] This study aimed to investigate the differences in morphological structure of tibias between sick and normal ostrich chicks. [Method] Normal and sick ostrich chicks aged 90 days were used in this research for bone mineral density determination, bone demineralization paraffin section preparation （HE staining）, bone abrasive disc preparation and scanning electron microscopy （SEM） observation, to analyze the differences in microstructure and ultrastructure of tibias between sick ostrich chicks and normal ones. [Result] Bone mineral density of ostrich chicks suffering from leg disease was much lower than that of the normal ones. Compared with normal tibias, less trabeculae were observed in sick tibias, which were attenuated and began to disappear. In addition, trabeculae micro-fracture could be observed under scanning electron microscope; resorption pits on bone slices formed by osteoclast could be observed under scanning electronic microscope. [Conclusion] These results indicate that the sick tibias all have lower bone density and trabeculae microfracture phenomenon in tibias with different diseases, which might be resulted from osteoporosis.
文摘In order to analyze skull variation in the genus Macaca ,seventy seven craniodental variables were taken from eleven species.They were first defined seven functional units comprising three anatomical regions.Twenty seven variables were finally selected to carry out the morphology of the whole skull.The data,organized in these ways,were examined to discover variations between and within the various species.The methods used were Principal Components Analysis (PCA) and Discriminant Function Analysis (DFA).PCAs of the functional units anatomical regions,and the whole skull provided similar,though not identical,separations of species clusters in both sexes separately.These differences in structure could be related to size,sexual dimorphism,diet,ecology,classification and phylogeny.The question of size should have been easy to settle.Unfortunately,this is not the case.In this study where the raw data are measurements of the specimens,the main differences should be size.However,the size differences seem to occur in both the first and second (independent) multivariate axes.In some analyses the size differences between the species are biggest and appear in the first axis.In other analyses it is the separation between the sexes (and these too are largely size) that are the biggest and appear in the first axis.Yet in other analyses,both of these size separations,though still orthogonal to one another,present in the combination of the first two axes.This certainly implies that a single axis of body size is not present and that shape differences have not been isolated form size differences.It also implies that sexual dimorphism is a complex matter.As a result,the question of the relationships between the species is therefore also complex.One cluster of species that includes M fascicularis,M sinica and M radiata was significantly isolated from all others regardless of level of analysis.This relationship is quite different from that proposed on the anatomy of the reproductive organs (Delson,1980;Fooden,1976,1980).
文摘Objective To Investigate stress adaptability of freeze-dried bone allograft.Methods Cortical and cancellous allograft were transplanted to each side of the midshaft diaphyseal ulna in two groups of 28 animals.The left transplanted allograft was free from fixation and bore a normal physiological lcad,while the right transplanted allograft was protected from loading by a simple external fixator and bore less load.Animals were sacrificed at the 2nd,4th,8th,16th week after transplantation and specimens were taken out for bone histomorphometry studies and analysis of collagen gene expression by in situ Cdna-Mrna hybridization.Results Labeled surface(LS)and bone mineral apposition rate(MAR)of the normally loaded graft-host bone interface were significantly higher than that of the less loaded side at the 4th,8th,16th week after transplantation.Parameters reflecting the internal repair process of the allograft,such as LS in cortical and cancellous bone or MAR in cortical bone of the normally loaded side were significantly higher than those of the less loaded side at the 16th week after transplantation.The result of in situ hybridization indicated that more osteoblast-like cells expressing the type Ⅰ collagen gene were found in the interface or interior of normally loaded grafts.Conclusion The stimulus of physiologic load can accelerate the early union of allograft-host bone interface and later new bone creep substitution to the necrotic allograft.
文摘Objective: To evaluate the osteogenic potential of bone morphogenetic protein (BMP)-2 gene transfected goat bone marrow-derived mesenchymal stem cells (MSCs). Methods: Goat bone marrow- derived MSCs were transfected by Adv-human bone morphogenetic protein (hBMP)-2 gene(Group 1), Adv-beta gal transfected MSCs (Group 2)and uninfected MSCs(Group 3). Western blot analysis, alkaline phosphatase staining, Von Kossa staining and transmission electron microscopy were adopted to determine the phenotype of MSCs. Then the cells were injected into thigh muscles of the nude mice. Radiographical and histological evaluations were performed at different intervals. Results: Only Adv-hBMP-2 transfected MSCs produced hBMP-2. These cells were positive for alkaline phosphatase staining at the 12th day and were positive for Von Kossa staining at the 16th day after gene transfer. Electron microscopic observation showed that there were more rough endoplasmic reticulum, mitochondria and lysosomes in Adv-hBMP-2 transfected MSCs compared to MSCs of other two groups. At the 3rd and 6th weeks after cell injection, ectopic bones were observed in muscles of nude mice of Group 1. Only fibrous tissue or a little bone was found in other two groups. Conclusions: BMP-2 gene transfected MSCs can differentiate into osteoblasts in vitro and induce bone formation in vivo.
