Study on the Intervention Effect of β-sitosterol in the Postmenopausal Osteoporosis / 浙江中医药大学学报

[Objective]To explore the intervention effect of β-sitosterol,an effective monomer component of Eucommia ulmoides,on postmenopausal osteoporosis in mice and its potential mechanism.[Methods]Twenty-four female C57BL/6J mice of 12 weeks old were randomly divided into sham operation group,model group and β-sitosterol intervention group,with 8 mice in each group.The model of postmenopausal osteoporosis was established by bilateral ovariectomy.Mice in sham group and model group were given 0.9％sodium chloride solution and β-sitosterol intervention group was given β-sitosterol.Eight weeks after operation,all the mice were sacrificed and samples were collected.Micro-computed tomography(Micro-CT)was used to observe the protective effect of the β-sitosterol intervention on postmenopausal bone loss and trabecular bone breakage.Hematoxylin-eosin(HE)staining was used to analyze the bone histomorphology of the distal femur.The tartrate-resistant acid phosphatase(TRAP)staining and the alkaline phosphatase(ALP)staining were used to evaluate bone resorption and bone formation respectively.Then the expression level of β-catenin in each group was detected by immunohistochemistry.[Results]Micro-CT scanning showed that β-sitosterol could reduce bone loss and bone microstructure destruction in postmenopausal mice.HE staining also confirmed that β-sitosterol could effectively maintain the number and morphology of trabecular bone and inhibit abnormal fat accumulation caused by estrogen deficiency.TRAP staining also showed that β-sitosterol treatment did not affect osteoclast formation,while immunohistochemical results showed that β-sitosterol could significantly promote the expression of osteogenic marker ALP.Moreover,β-sitosterol can increase the expression of β-catenin in the distal femur to promote bone formation.[Conclusion]β-sitosterol treatment could enhance osteogenic differentiation and promote bone formation to relieve postmenopausal osteoporosis by upregulating β-catenin expression in vivo.

Effect of Osteoking on Inflammatory Injury and NF-κB/NFATc1 Signaling Pathway in Model Mice with Postmenopausal Osteoporosis / 中国实验方剂学杂志

ObjectiveTo explore the possible mechanism of Osteoking （OK） on postmenopausal osteoporosis （PMOP）. MethodForty adult female mice were randomly divided into a sham operation （Sham） group， osteoporosis model （OVX） group， estradiol intervention （E2） group， and OK group， with 10 mice in each group. The modeling was completed by conventional back double incision ovariectomy， and the corresponding drugs were given one week later. After 12 weeks， the body mass and uterine index of mice were measured， and the pathological changes of bone tissue and the number of osteoclasts （OCs） were determined by hematoxylin-eosin （HE） and tartrate-resistant acid phosphatase （TRAP） staining， respectively. Bone mineral density （BMD）， trabecular number （Tb.N）， trabecular separation （Tb.Sp）， and bone volume fraction （BV/TV） were measured by microcomputed tomography （Micro-CT）. The maximum load of the femur was detected by a three-point bending test. The contents of tumor necrosis factor-α （TNF-α） and bone resorption marker C-terminal telopeptide of type Ⅰ collagen （CTX-1） were measured by enzyme linked immunosorbent assay （ELISA）. The protein expression levels of nuclear factor-kappa B p65 （NF-κB p65）， phosphorylated nuclear factor-kappa B p65 （p-NF-κB p65）， nuclear factor kappa B inhibitor alpha （IκBα）， phosphorylated nuclear factor kappa B alpha （p-IκBα）， nuclear factor of activated T cells 1 （NFATc1）， and proto-oncogene （c-Fos） were detected by Western blot. The mRNA expressions of OCs-related specific genes matrix metalloproteinase-9 （MMP-9）， NFATc1， TRAP， cathepsin K （CTSK）， and c-Fos were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the Sham group， the uterine index decreased significantly in the OVX group， and the body mass （BMI） increased significantly. The structure of bone trabeculae was completely damaged， and the number of OCs increased. BMD， Tb.N， BV/TV， and maximum load decreased， while Tb.Sp was up-regulated. The levels of TNF-α and CTX-1 in serum were up-regulated. The protein expressions of c-Fos， p-NF-κB p65/NF-κB p65， NFATc1， and p-IκBα/IκBα were increased. The mRNA expressions of NFATc1， c-Fos， CTSK， TRAP， and MMP-9 were up-regulated （P<0.05， P<0.01）. Compared with the OVX group， the body mass of the OK and E2 groups decreased， and the uterine index increased. The bone trabeculae increased， and the number of OCs decreased. BMD， Tb.N， BV/TV， and maximum load increased， while Tb.Sp decreased. The levels of TNF-α and CTX-1 in serum were decreased. The protein expressions of c-Fos， p-NF-κB p65/NF-κB p65， NFATc1， and p-IκBα/IκBα were decreased， and the mRNA expressions of NFATc1， c-Fos， CTSK， TRAP， and MMP-9 were decreased （P<0.05， P<0.01）. ConclusionOK can inhibit the NF-κB/NFATc1 signaling pathway and reduce bone mass loss by reducing the level of inflammatory injury factors in PMOP mice， which is one of the mechanisms for treating PMOP.

The role and mechanism of estrogen receptor in the treatment of postmenopausal osteoporosis by Gushukang / 中国组织工程研究

BACKGROUND:The specific mechanism of Gushukang,as a Chinese traditional patent medicine for the treatment of postmenopausal osteoporosis of kidney deficiency and blood stasis,needs further studies. OBJECTIVE:To investigate the effect of Gushukang on serum sex hormones,bone microstructure and estrogen receptor in postmenopausal osteoporosis. METHODS:Firstly,network pharmacological analysis was performed.The active ingredients and action targets of Gushukang and the targets of postmenopausal osteoporosis were obtained respectively.Cytoscape was used to construct the active ingredient-target network.STRING database and Cytoscape were used for protein-protein interaction analysis and screening of core targets.DAVID database was used for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of intersection targets.Then the ovariectomized Sprague-Dawley rats were used in the animal experiment.Gushukang was administered by gavage for 3 months.The serum estrogen level was detected by ELISA,the bone microstructure was detected by microCT,and the protein expression of estrogen receptor α and estrogen receptor β in bone tiusse was detected by western blot. RESULTS AND CONCLUSION:The network pharmacological research results identified 132 active ingredients and 150 targets of Gushukang and 1155 targets of postmenopausal osteoporosis.After intersections with 1155 postmenopausal osteoporosis targets,87 targets of active ingredients of Gushukang against postmenopausal osteoporosis were obtained.By constructing the active ingredient-target network,it was found that the active ingredients at the core were quercetin,kaempferol,luteolin,naringin and isorhamnetin,and the targets at the core were NCOA2,ESR2,AR,F2,ESR1 and PTGS1.The final targets obtained after the protein-protein interaction analysis and screening included MAPK8,ESR1,JUN,R3C1,RELA and FOS,of which ESR1 was the common core target obtained from the two analyses.KEGG enrichment analysis showed estrogen,tumor necrosis factor,apoptosis and other signaling pathways.Therefore,animal experiments focused on the effect of Gushukang on different subtypes of estrogen receptors in the estrogen signaling pathway.The results showed that in the Gushukang group,bone microstructure was significantly improved,serum estrogen level had no significant change,but the protein expression of estrogen receptor α and β in bone tissue was significantly increased.All the findings indicate that the mechanism of Gushukang in the treatment of postmenopausal osteoporosis may be related to its hormone-like effect and the enhancement of estrogen receptor expression.

Detection of immune-related cytokines of bone marrow mesenchymal stem cells in postmenopausal osteoporosis mice by antibody chip and analysis of key differential genes / 中国组织工程研究

BACKGROUND:In recent years,research on the interaction mechanism between the immune system and the skeleton in postmenopausal osteoporosis has become a hot topic.However,the impact of changes in key immune-related cytokine expression on postmenopausal osteoporosis remains unclear and requires further exploration. OBJECTIVE:To investigate the differential expression of immune-related cytokines in bone marrow mesenchymal stem cells of mice with postmenopausal osteoporosis by bioinformatics methods. METHODS:Postmenopausal osteoporosis mouse model was established through ovariectomy.Bone marrow mesenchymal stem cells were obtained by the whole bone marrow adherence method and passaged to passage 2.RayBio L-Series Mouse Antibody Array 308 Glass Slide Kit immune-related factor antibody chip was used to detect the differentially expressed proteins in bone marrow mesenchymal stem cells from ovariectomy and sham-operation mice.Gene ontology,Kyoto Encyclopedia of Genes and Genomes enrichment analysis,and protein-protein interaction network analysis were performed to screen common Hub genes by MCC,EPC,and MNC algorithms. RESULTS AND CONCLUSION:This study identified a total of 68 differentially expressed genes.Gene ontology analysis revealed that the differentially expressed genes were enriched in terms including"immune system processes","extracellular regions",and"signal receptor binding".Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the differentially expressed genes were mainly enriched in"cytokine-cytokine receptor interactions","tumor necrosis factor signaling pathways",and"chemokine signaling pathways".Further screening was performed by constructing a protein-protein interaction network analysis of these 68 differentially expressed genes to identify 8 Hub genes.The violin plot and correlation matrix showed that the expression levels of these 8 Hub genes were significantly down-regulated in the ovariectomy group compared to the sham-operation group.These results demonstrated that there was differential expression of immune-related factors in bone marrow mesenchymal stem cells of postmenopausal osteoporosis mice,and key genes involved in cytokine-cytokine receptor interactions,immune system-related processes,and potential targeted signaling pathways and cellular biological processes were identified,providing new promising targets for the diagnosis,treatment,and prevention of postmenopausal osteoporosis.

Punicalagin treats postmenopausal osteoporosis by promoting osteogenesis / 中国组织工程研究

BACKGROUND:Punicalagin has a wide range of effects and high safety,but its effect on osteoblasts and postmenopausal osteoporosis is unknown. OBJECTIVE:To investigate the effect of punicalagin on osteoblasts and postmenopausal osteoporosis. METHODS:The effect of punicalagin on the proliferation of MC3T3-E1 cells was detected.Punicalagin was added to the osteogenic induction medium to detect its effect on osteogenic differentiation.Punicalagin was used to treat ovariectomized rats and Micro CT scan and serum procollagen type 1 N-terminal propeptide test were performed after 3 months to detect the therapeutic effect. RESULTS AND CONCLUSION:Cell counting kit-8 assay showed that punicalagin could promote the proliferation of osteoblasts(P＜0.05).The results of qRT-PCR and western blot showed that punicalagin could promote the mRNA and protein expressions of alkaline phosphatase and Runx2 in osteoblasts(P＜0.05).The results of Micro CT scan and serological test showed that punicalagin could improve bone mineral density,bone volume fraction,trabecular thickness,trabecular number and procollagen type 1 N-terminal propeptide level of ovariectomized rats.To conclude,punicalagin can promote osteoblast proliferation and differentiation,and have therapeutic effects in postmenopausal osteoporosis rats.

Identification of biomarkers associated with ferroptosis and pyroptosis for the potential diagnosis of postmenopausal osteoporosis / 中国组织工程研究

BACKGROUND:Ferroptosis and pyroptosis may play a role in the development of postmenopausal osteoporosis.There may be relevant biomarkers for the diagnosis of postmenopausal osteoporosis. OBJECTIVE:To search for the key genes related to ferroptosis and pyroptosis in postmenopausal osteoporosis using bioinformatics so as to further elucidate their biological mechanisms. METHODS:The data sets GSE56815 and GSE7429 of postmenopausal osteoporosis were downloaded from the GEO database,the national comprehensive gene expression database of the United States,and the two data sets were preprocessed.The differential expression analysis of the data was carried out by the limma package of R software,and the enrichment analysis was performed by DIVID and KOBAS.The protein-protein interaction network was mapped by STRING and Cytoscape,the Hub gene was selected by CytoHubba,and the key genes were screened by the ferroptosis database and pyroptosis database.The CIBERSORT package was used to determine the immune infiltration of postmenopausal osteoporosis samples and to analyze the correlation between key genes and immune cells RESULTS AND CONCLUSION:A total of 30 differential genes of postmenopausal osteoporosis were screened in the experimental samples,of which 9 genes were up-regulated and 21 genes were down-regulated.The enrichment of GO and KEGG pathways showed that the differences were mainly in"serine-type endopeptidase activity,""innate immune response,""special particle lumen,"and"renin secretion."The protein-protein interaction network showed the correlation of differential genes and the top 10 Hub genes with"Degree"value were selected using CytoHubba.Hub gene was intersected with the FerrDb database and cell pyroptosis dataset to obtain key genes ELANE and LCN2.Receiver operating characteristic curve and box diagram showed that the expression of ELANE and LCN2 in serum samples of postmenopausal osteoporosis was significantly lower than that in normal samples,indicating a good diagnostic value.Immune infiltration analysis showed that ELANE may be related to memory resting CD4+ T cells,M0 and M2 macrophages.LCN2 may be related to M0 macrophages.

Mechanisms by which high-intensity interval training influences bone health in a rat model of postmenopausal osteoporosis / 中国组织工程研究

BACKGROUND:Resistance training and weight-bearing exercise are recommended modes for patients with osteoporosis to improve bone health.High-intensity interval training is a high-impact weight-bearing exercise with obvious time-efficient characteristics;however,little attention has been paid to its impact on bones. OBJECTIVE:To observe the effect of high-intensity interval training on the bone health of ovariectomized rat models. METHODS:Thirty-six female Sprague-Dawley rats were randomly divided into sham group,model group and model exercise group(n=12 per group).Bilateral ovariectomy was used to prepare an osteoporosis rat model in the latter two groups.Six weeks after modeling,the model exercise group was subjected to a high-intensity interval training on an electric treadmill at 90%peak running speed for 2 minutes and 50%peak running speed for 1 minute as one session,a total of nine sessions,3 days per week,for 6 weeks.Rats in the sham and model groups were raised quietly in the mouse cage during the same period.The relevant indexes were tested 48-72 hours after the final training. RESULTS AND CONCLUSION:Compared with the sham group,bone mineral density,maximal load,stiffness,elasticity,trabecular volume fraction,and trabecular number decreased(P<0.05),while trabecular separation increased(P<0.05);the level of irisin in the serum,gastrocnemius and femur decreased(P<0.05);the expression of peroxisome proliferator-activated receptor γ coactivator-1α protein and fibronectin type Ⅲ domain-containing protein 5 mRNA and protein in the gastrocnemius muscle decreased(P<0.05);the expression of type I collagen,Osterix,and Runx2 mRNA in the femur decreased(P<0.05);and the expression of anti-tartrate acid phosphatase,receptor activator of nuclear factor κB ligand,and osteoclast-associated receptor mRNA increased in the model group(P<0.05).Compared with the model group,bone mineral density,fracture load,maximal load,stiffness,elasticity,average trabecular thickness,and trabecular number increased(P<0.05),and trabecular separation decreased(P<0.05);the level of irisin in the serum,gastrocnemius and femur increased(P<0.05);the expression of peroxisome proliferator-activated receptor γ coactivator-1α protein and fibronectin type Ⅲ domain-containing protein 5 mRNA and protein in gastrocnemius increased(P<0.05);the expression of type I collagen,Osterix,and Runx2 mRNA in the femur increased(P<0.05);and the expression of anti-tartrate acid phosphatase,receptor activator of nuclear factor κB ligand,and osteoclast-associated receptor mRNA decreased in the model exercise group(P<0.05).To conclude,short-term high-intensity interval training may improve bone health of ovariectomized rats through up-regulating the irisin level.

Bibliometric and visual analysis of postmenopausal osteoporosis based on highly cited SCI papers / 中国组织工程研究

BACKGROUND:Bibliometrics and visual analyses based on thematic literature are particularly important for understanding the foundation and frontiers of postmenopausal osteoporosis research. OBJECTIVE:To perform bibliometric,citation,and visualization analyses of highly cited SCI papers in postmenopausal osteoporosis research over the last 20 years. METHODS:The top 100 highly cited papers on postmenopausal osteoporosis published between 2003 and 2022 included in SCI-EXPANDED catalog of the Web of Science database were obtained for bibliometric measure and visual analysis using CiteSpace software. RESULTS AND CONCLUSION:The top 100 highly cited papers have a total of 67 377 citations in the Web of Science Core Collection,with an annual average of 49.17 citations per paper.Postmenopausal osteoporosis research primarily involves medical,engineering,biological,and multidisciplinary fields.The subcategories are dominated by endocrinology and metabolism,and medicine:internal medicine.Stable and close cooperative network relationships have been formed globally.United States,University of California System,Cummings,and Steven R are the country,research institution,and author,respectively,with the most highly-cited publications.The frontiers of postmenopausal osteoporosis research mainly include calcium and vitamin D supplementation and fracture risk,clinical studies of bisphosphonates in the treatment of postmenopausal osteoporosis,atypical femur fracture,clinical studies of new drugs and sequential treatment of postmenopausal osteoporosis,predictors of fracture risk,mid-and long-term follow-up of osteoporotic vertebral compression fractures,genetic polymorphisms and hereditary factors,formulation and updating of clinical practice guidelines for postmenopausal osteoporosis.Large cohort studies,high-quality randomized controlled trials,systematic reviews,meta-analyses,and clinical practice guidelines are the great engines that drive the development of clinical research in postmenopausal osteoporosis.We should make efforts in the above areas to improve China's international influence in the field of osteoporosis.

The Relationship between Bone Mineral Densitometry and Visceral Adiposity Index in Postmenopausal Women / A relação entre a densitometria mineral óssea e o índice de adiposidade visceral em mulheres na pós-menopausa

Abstract Objective It was aimed to compare visceral adiposity index (VAI) levels in patients with normal bone mineral density (BMD), osteopenia, and osteoporosis. Methods One hundred twenty postmenopausal women (40 with normal BMD, 40 with osteopenia, and 40 with osteoporosis) between the ages of 50 to 70 years were included in the study. For females, the VAI was calculated using the formula (waist circumference [WC]/[36.58 + (1.89 x body mass index (BMI))]) x (1.52/High-density lipoprotein [HDL]-cholesterol [mmol/L]) x (triglyceride [TG]/0.81 [mmol/L]). Results The time of menopause from the beginning was similar in all groups. Waist circumference was found to be higher in those with normal BMD than in the osteopenic and osteoporotic groups (p = 0.018 and p < 0.001, respectively), and it was also higher in the osteopenic group than in the osteoporotic group (p = 0.003). Height and body weight, BMI, blood pressure, insulin, glucose, HDL-cholesterol, and homeostasis model assessment-insulin resistance (HOMA-IR) levels were similar in all groups. Triglyceride levels were found to be higher in the normal BMD group, compared with the osteoporotic group (p = 0.005). The level of VAI was detected as higher in those with normal BMD, compared with the women with osteoporosis (p = 0.002). Additionally, the correlation analysis showed a positive correlation between dual-energy X-ray absorptiometry (DXA) spine T-scores, WC, VAI, and a negative correlation between DXA spine T-scores and age. Conclusion In our study, we found higher VAI levels in those with normal BMD, compared with women with osteoporosis. We consider that further studies with a larger sample size will be beneficial in elucidating the entity.

Resumo Objetivo O objetivo foi comparar os níveis de índice de adiposidade visceral (IVA) em pacientes com densidade mineral óssea (DMO) normal osteopenia e osteoporose. Métodos Cento e vinte mulheres na pós-menopausa (40 com DMO normal 40 com osteopenia e 40 com osteoporose) com idades entre 50 e 70 anos foram incluídas no estudo. Para o sexo feminino o VAI foi calculado pela fórmula (circunferência da cintura [CC]/[36 58 + (1 89 x índice de massa corporal (IMC))]) x (1 52/lipoproteína de alta densidade [HDL]-colesterol [mmol/L]) x (triglicerídeo [TG]/0 81 [mmol/L]). Resultados O tempo de menopausa desde o início foi semelhante em todos os grupos. A circunferência da cintura foi maior naqueles com DMO normal do que nos grupos osteopênicos e osteoporóticos (p = 0 018 e p < 0 001 respectivamente) e também foi maior no grupo osteopênico do que no grupo osteoporótico (p = 0 003) . Altura e peso corporal IMC pressão arterial insulina glicose HDL-colesterol e os níveis de avaliação do modelo de homeostase-resistência à insulina (HOMA-IR) foram semelhantes em todos os grupos. Os níveis de triglicerídeos foram maiores no grupo DMO normal em comparação com o grupo osteoporótico (p = 0 005). O nível de VAI foi detectado como maior naquelas com DMO normal em comparação com as mulheres com osteoporose (p = 0 002). Além disso a análise de correlação mostrou uma correlação positiva entre a absorciometria de raios-X de dupla energia (DXA) nas pontuações T da coluna CC VAI e uma correlação negativa entre as pontuações T da coluna DXA e a idade. Conclusão Em nosso estudo encontramos níveis mais elevados de VAI naquelas com DMO normal em comparação com mulheres com osteoporose. Consideramos que novos estudos com maior tamanho amostral serão benéficos na elucidação da entidade.

Cost-utility analysis of denosumab and teriparatide in the treatment of postmenopausal osteoporosis in Chinese women / 中国药房

OBJECTIVE To evaluate the cost-effectiveness of denosumab and teriparatide in the treatment of postmenopausal osteoporosis in Chinese women， and provide reference for relevant decision-making. METHODS From the perspective of health system in China， Excel 2003 was used to establish Markov model， and cost-utility analysis was used to evaluate the cost- effectiveness of denosumab or teriparatide combined with Calcium carbonate D3 tablets in the treatment of postmenopausal osteoporosis in Chinese women. Pharmacotherapy effects were obtained with network meta-analysis， and cost and health utility value data were obtained from published literature. The model cycle was 1 year， and the simulation time limit was the patient’s lifetime. Univariate sensitivity analysis and probabilistic sensitivity analysis were used to evaluate the effects of model parameter changes on the robustness of the results. Through scenario analysis， the cost-effectiveness of domestic drug cost used as drug cost of terlipatide group was discussed； the influence of residual effects of teriparatide on the results and the cost-effectiveness of sequential use of desumamab after terlipatide withdrawal were also discussed. RESULTS The effect of denosumab regimen was better than that of terlipatide regimen [13.24 quality-adjusted life years （QALYs） vs. 12.96 QALYs]， with lower cost （51 224.64 yuan vs. 167 102.67 yuan）， denosumab regimen was the absolutely superior regimen. The results of single factor sensitivity analysis showed that the cost and discount rate of Terlipatide injection had greater impact on the results. The results of probability sensitivity analysis showed that when three times of China’s per capita gross domestic product （GDP） in 2021 was used as the threshold of willingness to pay， the probability of cost-effectiveness of denosumab regimen was 93.5%. The results of scenario analysis showed that， whether the drug cost of terlipatide regimen which was replaced by domestic drugs， or the residual effect of terlipatide was considered， or desulmonab was used sequentially after two years of terlipide treatment， denosumab regimen was always the absolute advantage regimen. CONCLUSIONS Denosumab combined with Calcium carbonate D3 tablets is more cost-effective than teriparatide combined with Calcium carbonate D3 tablets in the treatment of postmenopausal osteoporosis in Chinese women.

Mechanism of Zhibai Dihuang decotion in treating postmenopausal osteoporosis / 药学学报

In this study, the ovarian surgery (ovariectomy, OVX) was used to establish the osteoporosis mice model of primary menstruation, in order to evaluate the protective effects and mechanisms of Zhibai Dihuang decotion on postmenopausal osteoporosis (PMOP). The animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Jinan University (number: 20210315-03), in compliance with the Institutional Animal Care Guidelines. C57BL/6 mice were divided into five groups, including Sham group, OVX group, low (32 g·kg-1·day-1) and high dose (64 g·kg-1·day-1) of Zhibai Dihuang decotion groups, positive drug group (alendronate, 9.9 mg·kg-1·q3d). After modeling, mice were given medication intervention for 8 weeks, and then femoral and tibial tissues were taken to detect indicators such as bone microstructure, bone resorption, and oxidative stress. The experimental results showed that after Zhibai Dihuang decotion administration, the bone microstructure damage caused by OVX surgery was alleviated, and the relevant parameters bone mineral density (BMD), bone volume/total volume (BV/TV), trabecular number (Tb. N) and connectivity density (Conn. D) both significantly increased. At the same time, the number of TRAP positive osteoclasts decreased significantly, and the levels of proteins and genes related to osteoclast differentiation decreased, indicating that Zhibai Dihuang decoction could inhibit the increased activity of osteoclast caused by OVX. Afterwards, network pharmacology was used to construct the active compound action target network of Zhibai Dihuang decotion, and it was found that the target genes of its active ingredients were closely related to the oxidative stress pathway. Finally, the detection results of oxidative stress levels in bone tissues showed that after treatment with Zhibai Dihuang decotion, the levels of oxidative stress products 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA) in bone tissues of mice significantly decreased, while the levels of antioxidant stress substance L-glutathione (GSH) increased. These above results indicated that Zhibai Dihuang decotion can regulate the level of oxidative stress in the body and inhibit osteoclast activity, which played a therapeutic role in PMOP, as well as provided theoretical basis for the prevention and treatment of PMOP with traditional Chinese medicine.

Association of serum sRANKL and Omentin-1 levels with bone mineral density and bone metabolism in postmenopausal osteoporosis patients / 中华内分泌外科杂志

Objective:To investigate the relationship between serum soluble receptor activator of nuclear factor-κB ligand (sRANKL), Omentin-1 levels and postmenopausal osteoporosis (PMOP) .Methods:A total of 310 menopausal patients admitted to Qingdao Municipal Hospital from Jun. 2017 to Jul. 2021 were selected, including 165 patients with PMOP and 145 women with simple menopause as the control group. Serum sRANKL and Omentin-1 levels were detected by ELISA. Bone mineral density and bone metabolism indexes [N-terminal propeptide of typeⅠprecollagen (PINP), bone alkaline phosphatase (BALP), β isomer of the C-terminal telopeptide of type Ⅰ collagen (β-CTX) and osteocalcin (OC) ] were compared between the two groups. The correlation between serum sRANKL and Omentin-1 levels and bone mineral density and bone metabolism indexes in PMOP patients was analyzed by Pearson. The predictive value of sRANKL and Omentin-1 to PMOP was analyzed by ROC curve. Logistic regression analysis of the influence of multiple factors on PMOP.Results:Compared with the control group (15.62±4.41) (42.56±8.53), the serum sRANKL level (26.63±8.12) was increased and Omentin-1 level (32.32±5.52) was decreased in PMOP group ( t=14.55, P<0.001; t=12.69, P<0.001). The serum sRANKL in PMOP group was positively correlated with PINP, β-CTX and OC, while the serum Omentin-1 level was negatively correlated with the above indexes by Pearson analysis. ROC curve showed that serum sRANKL and Omentin-1 had important reference significance in predicting PMOP. Logistic regression suggested that increased sRANKL and decreased Omentin-1 were risk factors for PMOP. Conclusion:Serum sRANKL and Omentin-1 in patients with PMOP are correlated with bone mineral density and bone metabolism, and have potential as diagnostic targets of PMOP.

The Osteoprotective Effect of Gusong Yigu Decoction on Ovariectomized Rats Based on PI3K/Akt Signal Pathway / 中药新药与临床药理

Objective To explore the mechanism of Gusong Yigu Decoction(inchuding Astragali Radix,Codonpsis Radix,Angelicae Sinensis Radix,etc.)in the treatment of postmenopausal osteoporosis(PMOP)based on network pharmacology and animal experiment.Methods The effective ingredients and corresponding targets of Gusong Yigu Decoction were collected by using TCMSP database.GeneCards,TTD,and other databases were used to collect PMOP target proteins.R language was used to obtain the intersection targets and draw Wayne diagram.STRING database was used for the establishment of protein-protein interaction network.At last,GO function enrichment and KEGG pathway enrichment were performed on all common targets.The ovariectomized SD rats were used in the animal experiment.Gusong Yigu Decoction was administered by gavage for 12 weeks.The changes of bone histomorphology were detected by HE staining,the mRNA and protein levels of phosphatidylinositol 3-kinase(PI3K)and protein kinase B(Akt)in bone tissue of proximal tibial were tested by qRT-PCR and Western Blot,respectively.Results A total of 91 effective ingredients of Gusong Yigu Decoction in the treatment of PMOP,70 common targets of drugs-diseases were obtained.GO enrichment analysis mainly included DNA-binding transcription activator activity,RNA polymerase II-specific,ubiquitin protein ligase binding.KEGG pathway enrichment analysis included PI3K/Akt signaling pathway,TNF signaling pathway,and apoptosis.The animal experiment showed that bone histomorphology was significantly improved,meanwhile the mRNA and protein expressions of PI3K and Akt were significantly increased in Gusong Yigu Decoction group(P＜0.01).Conclusion Gusong Yigu Decoction may improve bone microstructure through multiple channels and targets.Gusong Yigu Decoction can increase the number and thickness of bone trabeculae and reduce the separation of bone trabeculae by activating PI3K/Akt signaling pathway,and thus play an anti-osteoporosis role in postmenopausal osteoporosis.

Iron accumulation and its impact on osteoporotic fractures in postmenopausal women / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Postmenopausal osteoporosis is a kind of degenerative disease, also described as "invisible killer." Estrogen is generally considered as the key hormone for women to maintain bone mineral content during their lives. Iron accumulation refers to a state of human serum ferritin that is higher than the normal value but less than 1000 μg/L. It has been found that iron accumulation and osteoporosis could occur simultaneously with the decrease in estrogen level after menopause. In recent years, many studies indicated that iron accumulation plays a vital role in postmenopausal osteoporosis, and a significant correlation has been found between iron accumulation and fragility fractures. In this review, we summarize and analyze the relevant literature including randomized controlled trials, systematic reviews, and meta-analyses between January 1996 and July 2022. We investigate the mechanism of the effect of iron accumulation on bone metabolism and discuss the relationship of iron accumulation, osteoporosis, and postmenopausal fragility fractures, as well as the main clinical treatment strategies. We conclude that it is necessary to pay attention to the phenomenon of iron accumulation in postmenopausal women with osteoporosis and explore the in-depth mechanism of abnormal bone metabolism caused by iron accumulation, in order to facilitate the discovery of effective therapeutic targets for postmenopausal osteoporosis.

Effects of denosumab on bone mineral density around proximal femoral prosthesis after total hip replacement in postmenopausal osteoporotic patients / 中国骨伤

OBJECTIVE@#To evaluate the effect of denosumab on bone mineral density around proximal femoral prosthesis after total hip arthroplasty(THA) in the postmenopausal osteoporotic patients.@*METHODS@#Fifty-four consecutive patients underwent unilateral primary THA were included in this retrospective study. Twenty-five patients received denosumab for osteoporosis as the treatment group, and the twenty-nine without denosumab were the control group. At 1 week, 3month, 6 months, and 12 months after THA, bone turnover markers and proximal femoral periprosthetic bone mineral density (BMD) were measured.@*RESULTS@#At 3, 6 and 12 months after operation, the level of TRACP-5b in the control group was significantly higher than that in the treatment group (P<0.05);the level of bone-specific alkaline phosphatase (BALP) between two groups showed significant difference in 12 months after operation (control group was higher than treatment group, P<0.05). The BMD of Gruen 1 and Gruen 7 decreased at 3, 6 and 12 months after operation compared with 1 week after operation. Comparing the treatment group and the control group, the differences of the the decrease of BMD in Gruen 1 and Gruen 7 were no significant at 3 months after surgery. In Gruen 1, Gruen 7 at 6 months after operation and Gruen 1, Gruen 7 at 12 months after operation, the decrease of BMD in the control group was significantly higher than that in the treatment group(P<0.05). It is suggested that desudumab could inhibit the loss of BMD after 6 months, and continuously show a protective effect on bone mass at 12 months after operation.@*CONCLUSION@#After THA in postmenopausal patients with osteoporotic femoral neck fracture, Desuzumab can reduce the loss of BMD around the proximal femoral prosthesis and effectively inhibit bone resorption.

Curative effect analysis of PVP combined with Zoledronic acid in the treatment of postmenopausal osteoporotic vertebral fracture (with video) / 中华内分泌外科杂志

Objective:To explore the efficacy of PVP combined with zoledronic acid in the treatment of postmenopausal osteoporotic vertebral fractures.Method:90 patients with postmenopausal osteoporotic vertebral fractures treated in our hospital from Jul. 2018 to Aug. 2020 were selected. According to different treatment methods, the patients were divided into observation group and control group. The patients in both groups were treated with PVP. The patients in the control group were given oral alendronate, calcitriol capsules and calcium, and the patients in the observation group were given zoledronic acid injection, calcitriol capsules and calcium were taken orally for 1 year. BMD of lumbar spine, femoral neck, greater trochanter and ward triangle, lumbar ODI index and VAS score, serum BGP and β-CTX, PINP levels and adverse reactions during treatment. Results:One year after operation, the lumbar ODI index and VAS score of the observation group were significantly lower than those of the control group ( P<0.05), and the BMD of lumbar spine, femoral neck, greater trochanter and ward triangle were higher than those of the control group ( P<0.05). The levels of serum BGP, β-CTX and PINP levels in the two groups were significantly higher than those in the control group ( P<0.05) . Conclusion:Zoledronic acid can significantly improve bone metabolism, accelerate bone formation, increase BMD, reduce low back pain and improve lumbar function in patients with osteoporotic vertebral compression fracture after PVP.

Association of serum NLR and SII with postmenopausal osteoporotic vertebral compression fractures and their predictive value for short-term prognosis / 中华内分泌外科杂志

Objective:To explore the correlation between serum NLR and SII levels and postmenopausal osteoporotic vertebral compression fracture (OVCF) and to analyze the short-term prognostic value.Methods:A total of 132 patients with postmenopausal OVCF admitted to our hospital from Dec. 2018 to Dec. 2021 were selected as the study group, and 98 patients with postmenopausal osteoporosis but did not suffer from OVCF were selected as the control group. According to the recurrence of postmenopausal OVCF fractures, the ROC curves of NLR and SII were plotted, and their prognostic value for postmenopausal osteoporosis OVCF was analyzed.Results:NLR level was 2.96±0.41 and STI level was 39.41±23.45 in the control group. The level of NLR was 3.42±0.32 and SII was 431.77±31.14 in the research group ( P<0.05) . Multivariate Logistic regression analysis showed that lumbar bone density ( OR=0.030, 95%CI: 0.001-0.832, P=0.042) , NLR level ( OR=29.43, 95%CI: 9.840-103.6, P=0.001) and SII level ( OR=1.048, 95%CI: 1.034-1.066, P=0.001) were all risk factors affecting postmenopausal OVCF. NLR (3.77±0.22) and SII (441.32±29.68) in the recurrent fracture group were higher than NLR (3.27±0.22) and SII (426.87±30.57) in the non-recurrent fracture group, and the differences were statistically significant (all P<0.05) , multivariate Logistic regression analysis showed lumbar spine bone density ( OR=8.56×10 4, 95% CI: 3.884-2.992×10 10, P=0.045) , NLR level ( OR=1.243×10 -8, 95% CI: 2.911×10 -13-1.072×10 -5, P=0.001) and SII level ( OR=0.938, 95% CI: 0.885-0.976, P=0.008) were all influencing factors affecting the postoperative treatment effect of postmenopausal OVCF, and ROC results showed that both NLR (AUC=0.86, 95% CI: 0.77-0.94, P<0.001) and SII (AUC=0.76, 95% CI: 0.67-0.85, P<0.001) had good prognostic value for postmenopausal OVCF. Conclusion:NLR and SII are risk factors for OVCF in postmenopausal osteoporosis patients, and have good short-term prognostic value.

Study on the Interventional Effects of Cornus Officinalis Before and After Wine Preparation in Liu Wei Di Huang Decoction on Postmenopausal Osteoporosis Model Rats Based on Medicinal / 世界科学技术-中医药现代化

Objective To establish a dual wavelength HPLC method to compare the content changes of seven active components:monoglycoside,loganin,swertin,paeoniflorin,gallic acid,5-hydroxymethyl furfural and paeonol in Liuwei Dihuang Decoction(hereinafter referred to as"Liuweidihuang Decoction")before and after the preparation of Cornus wine;To explore the effect and mechanism of Liu(Shan)and Liu(Jiu)on 90 days after ovariectomy in postmenopausal osteoporosis model rats.Methods ①The contents of seven active ingredients in the Cornus wine mixed with Liuwei Dihuang Decoction were determined by dual wavelength HPLC.Chromatographic conditions:hypersil C18 chromatography(4.6 mm×250 mm,5 μm),mobile phase acetonitrile-0.3%phosphoric acid,gradient elution(0-7 min,0%-8%acetonitrile;7-15 min,8%-10%;15-20 min,10%-15%;20-30 min,15%-23%;30-50 min,23%-45%),flow rate 0.6 mL·min-1,detection wavelength:240 nm(monoglycoside,loganin,swertin,paeoniflorin),274 nm(gallic acid,5-hydroxymethyl furfural,paeonol),column temperature 30℃,injection volume 20 μL.②6-8 month old SD rats,divided into sham-operated group,model group,positive drug group,shanshuang flesh with Liuwei Dihuang Decoction group(liu(shan)group)and wine cornelian flesh withLiuwei Dihuang Decoction group(liu(wine)group),dosing started on the 5 d after ovariectomy,weighed weekly,and 5 rats from each group were taken at 90 d after surgery,weighed,anaesthetized,serum taken,bilateral femurs,and organs(uterus,spleen,liver,both kidneys and heart)were weighed.Serum calcium(Ca),phosphorus(P)and estradiol(E2)levels were measured using a fully automated biochemical instrument and chemiluminescent microparticle immunoassay,respectively;bone mineral density(BMD)values were measured using a dual-energy X-ray bone densitometer.HE staining was used to observe the morphological changes of bone tissue in rats.Results ①All the seven potent ingredients mentioned above were contained in the Liuwei Dihuang Decoction before and after the wine preparation of Cornu Cervi Pantotrichum,and the total content of the seven potent ingredients in liu(wine)was higher than that in liu(shan)(P<0.01).Among them,gallic acid and 5-hydroxymethylfurfural were higher in Liu(Jiu)than in Liu(Shan)(P<0.01),while the content of monosidine was lower than in Liu(Shan)(P<0.01).②Comparison with the sham-operated group:90 d after surgery,the serum Ca,P,E2 levels and uterine index of rats in the model group decreased(P<0.01),the BMD value decreased(P<0.01),and the degree of structural damage to bone tissue The degree of damage to the bone tissue structure was deepened,manifested by enlargement of the bone marrow cavity,gradual increase of adipocytes,dilution of the bone trabeculae arrangement,reduction of the number of bone trabeculae or even fracture.③Compared with the model group,90 d after surgery,the serum E2,Ca,P levels and uterine index of the rats in the liu(shan)and liu(wine)groups increased(P<0.05),the bone tissue structure of the liu(shan)and liu(wine)groups improved significantly,and the BMD value increased significantly(The liver and spleen indices of the liu(wine)groups increased(P<0.05).Conclusion The combination of the wine preparation of Cornus officinalis and its use in the formula of LiuWei Di Huang Decoction both contain monosidine,strychnine,sweretin,paeoniflorin,gallic acid,5-hydroxymethyl furfural(5-HMF)and tannin,and can effectively interfere with PMOP.Among them,the anti-osteoporotic effect of Liu Wei Di Huang Tang,which was formulated with wine and dogwood,was better than that of Liu Wei Di Huang Tang,which was formulated with Cornus officinalis,and the former contained more 5-HMF and gallic acid than the latter,which may provide some experimental basis for the selection of wine and dogwood in the 2020 edition of the Pharmacopoeia and the clinical application of Liu Wei Di Huang Tang in the prevention and treatment of PMOP.

Factors affecting postmenopausal osteoporosis in Dali Bai Autonomous Prefecture / 预防医学

Objective@#To investigate the prevalence of postmenopausal osteoporosis (PMOP) and analyze its influencing factors among women at ages of 50 to 59 years in Dali Bai Autonomous Prefecture, Yunnan Province, so as to provide insights into the prevention of PMOP among menopausal women.@*Methods@#Bai Ethnic menopausal women at ages of 50 to 59 years who received healthy examination at the Center of Healthy Examination, Dali Prefecture People's Hospital from June 2017 to May 2021 were selected as the study subjects, and subjects' demographic characteristics, living habits, history of diseases, family history of osteoporosis and history of parturition were collected using self-designed questionnaires. The height, body weight and bone density were measured, and fasting blood glucose, vitamin D3, blood lipids and liver functions were detected. The factors affecting the development of PMOP were identified using a multivariable logistic regression model.@*Results@#Totally 2 000 questionnaires were allocated, and 1 584 valid questionnaires were recovered, with an effective recovery rate of 79.20%. The respondents had a mean age of ( 56.22±2.61 ) years, and mean body mass index ( BMI ) of ( 24.62±2.35 ) kg/m2. There were 497 respondents ( 31.38% ) with a family history of osteoporosis, and the prevalence of PMOP was 20.64%. Multivariable logistic regression analysis identified age ( OR=1.135, 95%CI: 1.074-1.196 ), age of menarche ( OR=1.138, 95%CI: 1.059-1.217 ), duration of menopause (OR=1.425, 95%CI: 1.228-1.622), number of parturition ( >2, OR=5.036, 95%CI: 2.972-7.101 ), smoking ( OR=2.594, 95%CI: 1.767- 3.421 ), alcohol consumption ( OR=2.051, 95%CI: 1.503-2.598 ), family history of osteoporosis ( OR=2.540, 95%CI: 1.769-3.311 ), hypertension ( OR=1.492, 95%CI: 1.406-1.578 ), diabetes ( OR=1.774, 95%CI: 1.581-1.967 ), total cholesterol ( OR=1.483, 95%CI: 1.251-1.716 ), triacylglycerol ( OR=1.801, 95%CI: 1.576-2.026 ), low-density lipoprotein cholesterol ( OR=1.614, 95%CI: 1.498-1.731 ), fasting blood glucose ( OR=1.192, 95%CI: 1.077-1.307 ), BMI ( OR=0.934, 95%CI: 0.862-0.993 ), outdoor activity ( ≥1 time/week, OR: 0.413-0.549, 95%CI: 0.329-0.637 ), age of menopause ( OR=0.909, 95%CI: 0.841-0.977 ), daily intake of calcium ( ≥600 mg, OR: 0.493-0.644, 95%CI: 0.389-0.786 ), vitamin D3 level ( ≥20 ng/mL, OR: 0.604-0.719, 95%CI: 0.523-0.853 ) and high-density lipoprotein cholesterol ( OR=0.658, 95%CI: 0.550-0.767 ) as factors affecting the development of PMOP.@*Conclusions @#The prevalence of PMOP in Dali Bai Autonomous Prefecture is similar to the nationwide level in China, and old age, smoking, alcohol consumption, a family history of osteoporosis and high blood lipid levels may increase the risk of PMOP.

Mechanism of Bushen Huatan Prescription in Regulation of Osteogenesis and Adipogenic Differentiation and Improvement of Postmenopausal Bone Loss in Ovariectomized Rats with Osteoporosis / 中国实验方剂学杂志

ObjectiveTo explore the underlying mechanism of Bushen Huatan prescription in alleviating postmenopausal osteoporosis （PMOP） by maintaining the balance of osteogenesis and adipogenic differentiation in ovariectomized rats with osteoporosis. MethodSeventy-five 6-month-old non-pregnant female SD rats were randomly divided into sham-operation group， model group， atorvastatin group， liviol group， and Bushen Huatan prescription group. Bilateral ovaries were removed in the four groups except the sham-operation group， while only the same mass of adipose tissue around the ovaries was removed in the sham-operation group. On the 5th week after surgery， drugs were consecutively administrated for 8 weeks. Rats in the Bushen Huatan prescription group received 9.4 mg·kg-1 of the prescription， rats in the atorvastatin group received 0.92 mg·kg-1 of atorvastatin， rats in the Liviol group received 0.23 mg·kg-1 of liviol， and rats in the model group and the sham-operation group received saline once a day. Micro-computed tomography （Micro CT） was used to detect bone mineral density （BMD） of rat tibia in each group. Hematoxylin-eosin （HE） staining was used to detect the relative area of rat bone marrow adipose tissue （BMAT） in each group. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the relative expression levels of Runt-related transcription factor 2 （Runx2）， peroxisome proliferator-activated receptor （PPARγ）， leptin （LPN）， and leptin receptor （OBR） in bone tissues. ResultAs compared with the sham operation group， the BMD of rats in the model group decreased （P<0.05）， while the relative area of BMAT increased （P<0.05）. In addition， the expression levels of LPN， OBR， and Runx2 decreased in the model group （P<0.05）， while the level of PPARγ increased （P<0.05）. As compared with the model group， the BMD of rats in the atorvastatin group， the Livial group， and the Bushen Huatan prescription group increased （P<0.05）， and the relative area of BMAT decreased （P<0.05）. The expression levels of LPN， OBR， and Runx2 in these groups increased （P<0.05）， while the expression level of PPARγ decreased （P<0.05）. ConclusionBushen Huatan prescription plays the anti-osteoporosis role in the rat model of PMOP through up-regulating LPN and OBR in bone tissues and maintaining the balance of osteogenesis and adipogenic differentiation， thereby reducing postmenopausal bone loss and playing a role in the prevention and treatment of PMOP.