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Type 2 diabetic osteoporosis (T2DOP) is a skeletal metabolic syndrome characterized by impaired bone remodeling due to type 2 diabetes mellitus, and there are drawbacks in the present treatment. Osteoking (OK) is widely used for treating fractures and femoral head necrosis. However, OK is seldom reported in the field of T2DOP, and its role and mechanism of action need to be elucidated. Consequently, this study investigated whether OK improves bone remodeling and the mechanisms of diabetes‐induced injury. We used db/db mice as a T2DOP model and stimulated MC3T3‐E1 cells (osteoblast cell line) with high glucose (HG, 50 mM) and advanced glycation end products (AGEs, 100 µg/mL), respectively. The effect of OK on T2DOP was assessed using a combined 3‐point mechanical bending test, hematoxylin and eosin staining, and enzyme‐linked immunosorbent assay. The effect of OK on enhancing MC3T3‐E1 cell differentiation and mineralization under HG and AGEs conditions was assessed by an alkaline phosphatase activity assay and alizarin red S staining. The AGEs/insulin‐like growth factor‐1(IGF‐1)/β‐catenin/osteoprotegerin (OPG) pathway‐associated protein levels were assayed by western blot analysis and immunohistochemical staining. We found that OK reduced hyperglycemia, attenuated bone damage, repaired bone remodeling, increased tibial and femoral IGF‐1, β‐catenin, and OPG expression, and decreased receptor activator of nuclear kappa B ligand and receptor activator of nuclear kappa B expression in db/db mice. Moreover, OK promoted the differentiation and mineralization of MC3T3‐E1 cells under HG and AGEs conditions, respectively, and regulated the levels of AGEs/IGF‐1/β‐catenin/OPG pathway‐associated proteins. In conclusion, our results suggest that OK may lower blood glucose, alleviate bone damage, and attenuate T2DOP, in part through activation of the AGEs/IGF‐1/β‐catenin/OPG pathway. [ABSTRACT FROM AUTHOR]

Background: Osteoking has been extensively used for the treatment of knee osteoarthritis (KOA). However, it is lack of high-quality evidence on the clinical efficacy of Osteoking against KOA and the comparison with that of nonsteroidal anti-inflammatory drugs (NSAIDs). Aims: To evaluate the efficacy and safety of Osteoking in treating KOA. Methods: In the current study, a total of 501 subjects were recruited from 20 medical centers, and were divided into the Osteoking treatment group (n = 428) and the NSAIDs treatment group (n = 73). The Propensity Score Matching method was used to balance baseline data of different groups. Then, the therapeutic effects of Osteoking and NSAIDs against KOA were evaluated using VAS score, WOMAC score, EQ-5D-3L and EQ-VAS, while the safety of the two treatment were both assessed based on dry mouth, dizziness, diarrhea, etc. Results: After 8 weeks of treatment, the Osteoking group was compared with the NSAIDs group, the VAS score [2.00 (1.00, 3.00) vs. 3.00 (2.00, 4.00)], WOMAC pain score [10.00 (8.00, 13.00) vs. 11.00 (8.00, 16.00) ], WOMAC physical function score [32.00 (23.00, 39.00) vs. 39.07 ± 16.45], WOMAC total score [44.00 (31.00, 55.00) vs. 53.31 ± 22.47) ], EQ-5D-3L score [0.91 (0.73, 0.91) vs. 0.73 (0.63, 0.83) ] and EQ-VAS score [80.00 (79.00, 90.00) vs. 80.00 (70.00, 84.00) ] were improved by the treatment of Osteoking for 8 weeks more effectively than that by the treatment of NSAIDs. After 8 weeks of treatment with Osteoking, the VAS scores of KOA patients with the treatment of Osteoking for 8 weeks were reduced from 6.00 (5.00, 7.00) to 2.00 (1.00, 3.00) (p < 0.05), which was better than those with the treatment of NSAIDs starting from 2 weeks during this clinical observation. Importantly, further subgroup analysis revealed that the treatment of Osteoking was more suitable for alleviating various clinical symptoms of KOA patients over 65 years old, with female, KL II-III grade and VAS 4-7 scores, while the clinical efficacy of NSAIDs was better in KOA patients under 65 years old and with VAS 8-10 scores. Of note, there were no differences in adverse events and adverse reactions between the treatment groups of the two drugs. Conclusion: Osteoking may exert a satisfying efficacy in relieving joint pain and improving life quality of KOA patients without any adverse reactions, especially for patients with KL II-III grades and VAS 4-7 scores. [ABSTRACT FROM AUTHOR]

Background: Osteoking (OK) is prescribed in traditional Chinese medicine to accelerate fracture healing. Although some studies suggest the potential efficacy of OK for fracture healing, the evidence remains inconclusive. Aim: To systematically evaluate the safety of OK and its effect on fracture healing. Methods: Relevant authoritative databases were searched until 25 August 2023. Randomized controlled trials (RCTs) of patients with fractures treated with Osteoking were included. We evaluated the risk of bias using the Cochrane tool and performed a meta-analysis using the Review Manager 5.4 software package. Results: 13 studies involving 1123 participants were included. This meta-analysis showed that compared with observations in the control group, theOK group showed a shortened fracture healing time, increased fracture healing rate, reduced swelling regression time and ecchymosis regression time, and improved bone metabolism. In addition, the included studies did not report any serious side effects associated with the use of OK, and the mild side effects resolved without treatment. Conclusion: OK therapy is beneficial and safe for accelerating fracture healing, reducing swelling, eliminating ecchymosis, and improving bone metabolism. However, the meta-analysis results do not support OK treatment for improving the fracture healing rate at all fracture sites and reducing pain across all fracture sites. Further original, high-quality studies are needed to validate these findings. [ABSTRACT FROM AUTHOR]

Objective To investigate the potential mechanism of osteoking in the treatment of type 2 diabetic osteoporosis by inhibiting osteoblast ferroptosis from the perspective of network pharmacology and molecular docking.Methods TCMSP database and BATMAN-TCM database were used to retrieve the components and targets of traditional Chinese medicine, and GeneCards and OMIM database were used to obtain the disease targets of osteoporosis in type 2 diabetes.The PPI protein interaction network was constructed with STRING11.0 database, and topological analysis was performed to screen important targets in the network. The potential therapeutic targets were imported into the DAVID online database for gene ontology(GO) enrichment analysis and gene and genome encyclopedia(KEGG) pathway enrichment analysis.Finally, molecular docking simulation was performed to verify the results of network pharmacology.Results A total of 223 potential therapeutic targets were obtained through network pharmacology screening.The key active substances of osteoking in the treatment of type 2 diabetic osteoporosis are luteolin, 20(R)-ginsenoside Rh2, kaempferol, baicalein, beta-sitosterol, etc.The key targets are IL-6, Akt, TNF, MAPK3, HIF-1A, etc.The results of GO and KEGG analysis showed that the treatment of osteoporosis in type 2 diabetes by osteoking mainly included biological processes such as lipopolysaccharide response, hypoxia response, and inflammatory response, and regulated HIF-1A, TNF, IL-17, PI3K/Akt and other signaling pathways.The results of molecular docking showed that 7-O-methylisoprostol, baicalein, kaempferol, and quercetin had good binding activity with HIF-1A.Conclusion Through network pharmacology and molecular docking technology, it is found that osteoking may treat type 2 diabetic osteoporosis through multiple targets, and may reduce oxidative stress and inhibit osteoblast ferroptosis through HIF-1A signaling pathway to treat type 2 diabetic osteoporosis. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The aim of this study was to predict the mechanism of Osteoking in preventing deep vein thrombosis (DVT) of the lower limb by network pharmacology and molecular docking. MATERIALS AND METHODS: The relevant active components and targets of Osteoking were collected through the TCMSP database, and the relevant disease targets of DVT were collected through the GeneCards, OMIM, and DisGeNET databases. The intersecting gene targets of Osteoking and DVT were obtained using Venny 2.1.0 software. PPI network construction and core target selection using Cytoscape 3.9.0 software. The Metascape database was used for GO and KEGG enrichment analysis of relevant targets. Finally, the molecular docking of the main active components and key targets was carried out. RESULTS: There are 361 potential targets and 71 core targets of Osteoking in preventing deep vein thrombosis of the lower limb. Signal pathways are involved in various diseases such as cancer, diabetic complications, atherosclerosis, and more. Some of the most common pathways include AGE-RAGE signaling pathway and Calcium signaling pathway. Molecular docking results showed that the main active components of Osteoking had relatively stable binding activities with the key targets. CONCLUSIONS: Osteoking can play a role through multiple targets and multiple signal pathways to prevent the formation of deep venous thrombosis of the lower limb after fracture. [ABSTRACT FROM AUTHOR]

Objective To explore the potential mechanism of Osteoking （OK） in the treatment of Osteoarthritis （OA） and preliminarily verify it with DMM rats. Methods TCMSP database was used to collect potential targets of OK, GeneCards database and OMIM were used to screen target genes related to OA, and Cytoscape 3.8.2 software was used to establish a regulatory network for interaction between OK Active ingredient and OA target genes. The STRING database was used to construct protein interaction maps and GO and KECG were analyzed. In addition, OA rats were constructed using medial meniscus instability surgery （DMM） to observe knee joint cartilage lesions, and ELISA was used to detect inflammation and oxidative stress levels after OK treatment. Results Network pharmacology results showed that 80 core genes were involved in OK therapy for OA, including serum albumin （ALB） and interleukin-6 （IL6), mainly involving biological processes such as immunity, inflammatory response, oxidative stress, and metabolism. Animal experiments showed that compared with the model group, the surface structure of the articular cartilage in the OK group of rats tended to be normal, with less detachment of chondrocytes; OK reduced serum inflammatory factors IL-6 （P < 0.01), VEGF （P < 0.01), and oxidative stress factor MDA （P < 0.05), improved ALB levels and SOD activity （P < 0.01). Conclusion OK can alleviate OA cartilage tissue damage, enhance immunity, and inhibit inflammation and oxidative stress reactions, preliminarily verifying the results of network pharmacology. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of Osteoking combined with antibiotic cocktail on insulin resistance and gut microbiota in db/db mice. Methods: The wild mice were used as the control group, while db/db mice were randomly divided into the model group, Osteoking group (OK), antibiotic group ( ABX), and Osteoking combined with antibiotic cocktail group (OA). After the intragastric administration for 11 weeks, the following indices were investigated: body weight, fasting blood-glucose (FBG), serum insulin and the changes of intestinal microflora by 16S rDNA sequencing technology in mice. Results: Compared with the model group, levels of FBG and HOMA- IR were significantly decreased in OK, ABX, AO groups (P < 0.05). And the shannon, simpson and pielou_e index of gut microbiota were significantly decreased in ABX group (P < 0.05), while they were increased in OK group (P < 0.05). The shannon and pielou_e index were increased in AO group (P < 0.05). At the phylum level, the relative abundance (RA) of Firmicutes (P < 0.05) in OK group, RA of Proteobacteria in ABX (P < 0.01）and AO groups (P < 0.01) were significantly increased; while RA of Bacteroidota was significantly decreased in OK (P < 0.05), ABX (P < 0.01), AO (P < 0.01）groups. At the family level, RA of Lachnospiraceae (P < 0.05) in OK group, Enterobacteriaceae, Sutterellaceae in ABX and AO groups (P <0.01）and Tannerellaceae in AO groups (P < 0.01) were significantly increased; while RA of Muribaculaceae in OK group was significantly decreased (P < 0.05). The following gut microbiota were riched: Proteobacteria in ABX group, Lachnospiraceae NK4A136 group, Lactobacillus, Desulfobacteraceae and Prevotellaceae in OK group, Parabacteroides gordonii, Akkermansia, Morganellaceae and Alphaproteobacteria in AO group. Conclusion: Osteoking, antibiotic cocktail alone or in combination have the effects of improving the insulin resistance in db/db mice. And Osteoking can improve the intestinal microflora imbalance induced by antibiotic cocktail. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To probe into the application value of osteoking after percutaneous vertebroplasty/ percutaneous kyphoplasty (PKP) for osteoporotic vertebral compression fracture (OVCF), and its effects on bone metabolism and bone mineral density of patients. METHODS: A total of 66 patients with OVCF undergoing PKP surgery in our hospital from Jun. 2019 to Jun. 2020 were extracted to be divided into the control group and the observation group via the random number table, with 33 cases in each group. The control group was treated with standard anti-osteoporosis scheme after surgery, while the observation group received osteoking on the basis of the control group, both groups were treated for 12 weeks. The efficacy, time of fracture healing were compared between two groups. Changes of bone metabolism and bone mineral density were observed before and after treatment, and the new fractures were followed up. RESULTS: The total effective rate of the observation group was 100. 00%, higher than that of the control group (87. 88%, 29/33),and the time of fracture healing of the observation group was shorter than that of the control group, with statistically significant differences (P <0. 05). After treatment, the levels of serum alkaline phosphatase (ALP) and osteocalcin (BGP) in both groups increased compared with before treatment, while the urinary tartrate-resistant acid phosphatase (U-TRAP), urinary calcium/ creatinine ( U-Ca2+ / Cr) and urinary hydroxyproline/ creatinine (U-HYP / Cr) decreased compared with before treatment;after treatment, the levels of ALP and BGP in the observation group were higher than those in the control group, while U-Trap, U-Ca2+ / Cr and U-HYP / Cr in the observation group were lower than those in the control group, the differences were statistically significant (P<0. 05). After treatment, the bone mineral density of lumbar spine L2—4 and femoral neck of two groups increased compared with before treatment, and the bone mineral density of lumbar spine L2—4 and femoral neck of the observation group was higher than that of the control group, the differences were statistically significant (P<0. 05). The rate of new fracture in the observation group was 3.03%(1/33), significantly lower than 18.18% (6/33) in the control group, with statistically significant differences (P<0. 05). CONCLUSIONS: Osteoking can significantly promote the bone formation, inhibit the bone resorption and enhance the bone mineral density in patients with OVCF, which is of positive significance to promote early fracture healing and prevent recurrent vertebral fractures. [ABSTRACT FROM AUTHOR]

Objective To explore the active ingredients, targets and mechanisms of Osteoking(OK, Herbs of the Yi Minority in Yunnan Province) in the treatment of osteoporosis (OP) with the help of systematic pharmacology. And the molecular docking technology is used for docking verification. Methods Screened the active components and targets of OK through TCMSP. Queried GeneCard, OMIM, DisGeNet, TTD Databases to Screening of OP-related gene targets. Constructed the intersection gene target protein-protein interaction(PPI) network by using STRING and Cytoscape Software.The core targets were analyzed by GO and KEGG with the help of DAVID and KEGG Mapper database.Finally, Autodock was used for docking verification. Results The main active components such as quercetin, kaempferoland,11-octadecenoic acid,11-eicosenoic acid,ursolic acid,beta-sitosterol,tormentic acid were collected. It involves biological processes such as cell responses to lipids, toxic substances, lipopolysaccharides, bacterial molecules, inorganic substances, organic ring compounds, organic nitrogen compounds and so on.Theymight play rolesthrough PI3K-Akt, Ras, MAPK, ect multiple pathways.Molecular docking suggested that ursolic acid had better binding effect with AKT1, ALB and IL-6.Conclusion It is preliminarily clear that OK participates in promoting bone formation and inhibiting bone fracture through multi-targets and multi-pathways, which provides theoretical support and design ideas for further experimental research. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is a common disease characterized by cartilage degeneration. In recent years much attention has been paid to Traditional Chinese Medicine (TCM) since its treatments have shown efficacy for ameliorating cartilage degradation with mild side effects. Osteoking is a TCM prescription that has long been used in OA treatment. However, the exact mechanism of Osteoking are not fully elucidated. In the current study, destabilization of the medial meniscus (DMM)-induced OA mice was introduced as a wild type animal model. After 8 weeks of administration of Osteoking, histomorphometry, OARSI scoring, gait analysis, micro-CT, and immunohistochemical staining for Col2, MMP-13, TGFβRII and pSmad-2 were conducted to evaluate the chondroprotective effects of Osteoking in vivo. Further in vitro experiments were then performed to detect the effect of Osteoking on chondrocytes. TGFβRIICol2ER transgenic mice were constructed and introduced in the current study to validate whether Osteoking exerts its anti-OA effects via the TGF-β signaling pathway. Results demonstrated that in wild type DMM mice, Osteoking ameliorated OA-phenotype including cartilage degradation, subchondral bone sclerosis, and gait abnormality. Col2, TGFβRII, and pSmad-2 expressions were also found to be up-regulated after Osteoking treatment, while MMP-13 was down-regulated. In vitro, the mRNA expression of MMP-13 and ADAMTS5 decreased and the mRNA expression of Aggrecan, COL2, and TGFβRII were up-regulated after the treatment of Osteoking in IL-1β treated chondrocytes. The additional treatment of SB505124 counteracted the positive impact of Osteoking on primary chondrocytes. In TGFβRIICol2ER mice, spontaneous OA-liked phenotype was observed and treatment of Osteoking failed to reverse the OA spontaneous progression. In conclusion, Osteoking ameliorates OA progression by decelerating cartilage degradation and alleviating subchondral bone sclerosis partly via the TGF-β signaling pathway.

Osteoarthritis (OA) is a common disease characterized by cartilage degeneration. In recent years much attention has been paid to Traditional Chinese Medicine (TCM) since its treatments have shown efficacy for ameliorating cartilage degradation with mild side effects. Osteoking is a TCM prescription that has long been used in OA treatment. However, the exact mechanism of Osteoking are not fully elucidated. In the current study, destabilization of the medial meniscus (DMM)-induced OA mice was introduced as a wild type animal model. After 8 weeks of administration of Osteoking, histomorphometry, OARSI scoring, gait analysis, micro-CT, and immunohistochemical staining for Col2, MMP-13, TGFβRII and pSmad-2 were conducted to evaluate the chondroprotective effects of Osteoking in vivo. Further in vitro experiments were then performed to detect the effect of Osteoking on chondrocytes. TGFβRIICol2ER transgenic mice were constructed and introduced in the current study to validate whether Osteoking exerts its anti-OA effects via the TGF-β signaling pathway. Results demonstrated that in wild type DMM mice, Osteoking ameliorated OA-phenotype including cartilage degradation, subchondral bone sclerosis, and gait abnormality. Col2, TGFβRII, and pSmad-2 expressions were also found to be up-regulated after Osteoking treatment, while MMP-13 was down-regulated. In vitro , the mRNA expression of MMP-13 and ADAMTS5 decreased and the mRNA expression of Aggrecan, COL2, and TGFβRII were up-regulated after the treatment of Osteoking in IL-1β treated chondrocytes. The additional treatment of SB505124 counteracted the positive impact of Osteoking on primary chondrocytes. In TGFβRIICol2ER mice, spontaneous OA-liked phenotype was observed and treatment of Osteoking failed to reverse the OA spontaneous progression. In conclusion, Osteoking ameliorates OA progression by decelerating cartilage degradation and alleviating subchondral bone sclerosis partly via the TGF-β signaling pathway. [ABSTRACT FROM AUTHOR]

Abstract Background Bone damage is a condition that affects the quality of life of patients. Mesenchymal stem cells (MSCs) are important for bone repair. Osteoking is a natural compound in traditional Chinese Medicine used to treat bone diseases; however, the effect of Osteoking on the differentiation of MSCs has not been reported. In this study, we aimed to investigate the effect of Osteoking on the osteogenic and adipogenic differentiation potential of rat bone marrow mesenchymal stem cells (rbMSCs). Methods The effects of Osteoking on the proliferation and differentiation of rbMSCs were investigated. Different concentrations of Osteoking were prepared, and its cytotoxicity was evaluated by CCK-8 assay. The expression of osteogenic and adipogenic genes were determined, and several staining methods were used to reveal the osteogenic and adipogenic differentiation potential of rbMSCs. Results Our results show that appropriate concentrations of Osteoking can enhance osteogenic differentiation of rbMSCs and reduce adipogenic differentiation without any effect on proliferation. This may be related to the changes in related gene expression. Conclusion Osteoking enhances osteogenic differentiation and inhibits adipogenic differentiation of rbMSCs. Therefore, Osteoking may have a therapeutic potential for treating bone disease caused by changes in differentiation function of MSCs.

Objective To observe the effect of osteoking on bone density and bone biomechanics in a postmenopausal osteoporotic fracture (OPF) model tree shrew. Methods Female western Yunnan subspecies tree shrews were subjected to bilateral ovariectomy and hysterectomy to establish an osteoporosis (OP) model, and after 180 days, bone density (BMD) was measured in tree shrews, and 20 OP modeling successes were randomly and equally divided into administration (3 mL/kg) and model groups (equal volume of 0.9% sodium chloride solution). Ten tree shrews with the same volume of large omentum removed were divided into normal groups (equal volume of 0.9% sodium chloride solution). The tree shrews in the drug-administered and model groups were subjected to right hindlimb tibial fracture surgery to establish the OPF model. The administration group was given one dose of saline every 2 d for 90 d, and the remaining group was given the same dose of saline. Results Compared with the normal group, the BMD of whole body (including head)，lumbar + pelvic, cervical-caudal, left pelvis, right pelvis, lumbar spine, lumbar + sacral, and whole body (excluding head) were significantly reduced to different degrees in the administered and model groups, and the differences were statistically significant (P < 0.05). Compared with the model group, the BMD of the whole body (including the head)，lumbar + pelvic, cervical-caudal, left pelvis, lumbar spine, and lumbar + sacral were significantly increased in the administration group, and the differences were statistically significant (P < 0.05), while the BMD of the right pelvis and whole body (excluding the head) were not statistically different. In terms of bone biomechanics, energy absorption was significantly increased in the administered group compared to the normal group, while maximum load and structural stiffness were not statistically difference; the maximum load, structural stiffness, and energy absorption capacity of the model group were all significantly reduced, and the differences were statistically significant (P < 0.05). The maximum load, structural stiffness, and energy absorption capacity of the model group were all significantly reduced compared to the drug-administered group, and the differences were statistically significant ( P < 0.05). Conclusion Osteoking can improve both BMD and bone biomechanics in the OPF model of tree shrew. [ABSTRACT FROM AUTHOR]

Objective To evaluate the effect of Osteoking on CD34 and VEGF in kidney of osteoporosis tree shrew. Methods A total of 30 12-month-old female tree shrew （Tupaia belangeri） were randomly divided into control group，model group and treatment group，with 10 in each group. Bilateral ovaries and uterus in the model group and treatment group were removed and osteoporosis was confirmed 6 months later. The treatment group were gavaged with Osteoking once every two days for 3 months，and others were gave with equal volume of saline. Animals were perfused for fixation and collected samples，and IHC mothed was employed to determine the protein expression of CD34 and VEGF in kidney. Results The protein expression of CD34（P < 0.05） and VEGF （P < 0.05） in the model group were significantly higher than the control group after 3 months. Compared with the model group， the protein expression of CD34（P < 0.05） and VEGF（P < 0.05） in the treatment group were significantly decreased. Conclusion Osteoking can effectively protect the kidney by regulating the overexpression of CD34 and VEGF in os-teoporosis castrated tree shrew. [ABSTRACT FROM AUTHOR]

Objective To evaluate the effect of Henggu bone wound healing agent on the white blood cell line, spleen coefficient, body weight and bone turnover markers of female tree shrews with osteoporotic fractures. Methods The experimental tree shrews were divided into 3 groups (control group, model group and treatment group), with 10 animals in each group. In the control group, only part of the adipose tissue of both ovaries was removed, and the treatment group and model group were removed from both ovaries. The experimental tree was bred for 6 months after castration, and the bone density was measured to confirm that the osteoporotic tree was successfully modeled and the tibia fracture operation was performed. The treatment group was given Henggu bone wound healing agent once/2 days; the control group and the model group were given the same amount of normal saline. After 3 months of administering, the tree white cell line was tested, the spleen coefficient was calculated, the body weight was weighed, and the bone turnover markers were tested. Results The white blood cell coefficient of the model group was lower than that of the control group (P ＜0.05). The absolute value and percentage of monocytes in the treatment group were higher than the model group (P ＜0.01). The number of white blood cells and lymphocytes in the treatment group were all higher In the model group (P<0.05). The spleen coefficient of the model group was extremely lower than that of the control group (P <0.01), and the spleen coefficient of the treatment group was significantly higher than that of the model group (P <0.05). The tree weight of the model group was extremely high than that of the control group (P <0.01), and the tree weight of the treatment group was extremely lower than that of the model group (P <0.01). Model group tree serum osteogenic markers osteocalcin (OC), type I procollagen N-terminal propeptide (PINP) and bone resorption markers type I collagen cross-linked carboxy-terminal peptide (CTX), anti-tartrate acid phosphatase ( TRACP-5b) were higher than the control group (P <0.01). OC, PINP, CTX, TRACP-5b in the treatment group were significantly lower than the model group (P <0.01). Conclusion Henggu bone healing agent can improve immunity and regulate bone remodeling of osteoporotic fracture trees. [ABSTRACT FROM AUTHOR]

Background: Bone damage is a condition that affects the quality of life of patients. Mesenchymal stem cells (MSCs) are important for bone repair. Osteoking is a natural compound in traditional Chinese Medicine used to treat bone diseases; however, the effect of Osteoking on the differentiation of MSCs has not been reported. In this study, we aimed to investigate the effect of Osteoking on the osteogenic and adipogenic differentiation potential of rat bone marrow mesenchymal stem cells (rbMSCs). Methods: The effects of Osteoking on the proliferation and differentiation of rbMSCs were investigated. Different concentrations of Osteoking were prepared, and its cytotoxicity was evaluated by CCK-8 assay. The expression of osteogenic and adipogenic genes were determined, and several staining methods were used to reveal the osteogenic and adipogenic differentiation potential of rbMSCs. Results: Our results show that appropriate concentrations of Osteoking can enhance osteogenic differentiation of rbMSCs and reduce adipogenic differentiation without any effect on proliferation. This may be related to the changes in related gene expression. Conclusion: Osteoking enhances osteogenic differentiation and inhibits adipogenic differentiation of rbMSCs. Therefore, Osteoking may have a therapeutic potential for treating bone disease caused by changes in differentiation function of MSCs. [ABSTRACT FROM AUTHOR]

Objective To study the effect of OK on the differentiation of bone marrow stromal stem cells （BMSCs） through the TGFβ/Smad signaling pathway in rats. Methods We removed the bilateral ovaries of rats to replicate OP model，and isolated and cultured BMSCs. BMSCs isolated from nomal rats were cultured as control group，BMSCs isolated from OP rats were divided into 5 groups，OP model group was regularly cultured，positive control group was treated with alendronate sodium（1 μmol/L），low，medium and high OK treatment group were treated with 50 mL/L，100 mL/L and 200 ml/L OK respectively．After 24 h incubation, all cells were collected. The proliferation rate of BMSCs was determined by MTT method，and ELISA method was employed to detect the contents of bone formation markers，RT-qPCR was used to determine the mRNA expression level of TGFβ and the protein expression levels of TGFβ，p-Smad2/3 and of Smad2/3 were detected by Western blot. Results Compared with control group，the proliferation rate of the BMSCs in OP model group was reduced，concentrations of bone formation markers （OC，PINP and BALP） were reduced，mRNA and protein expression levels of TGFβ，as well as the phosphorylated level of Smad2/3 were downregulated．The difference was statistically significant （P ＜0.05） . After treatment with OK，compared with model group，all the above effects were ameliorated in different degree，a dose dependent manner was observed in OK treatment group，and the treatment effects of alendronate sodium（1 μmol/L），100 mL/L and 200 mL/L OK group were statistically significant （P ＜0.05） .Conclusion OK can promote the proliferation of osteoblasts by activating TGFβ/Smad signaling pathway to achieve the effect of treating postmenopausal OP. [ABSTRACT FROM AUTHOR]

Objective To observe the effect of Osteoking on blood biochemicalindicator of tree shrews with osteoporotic fracture （OPF） ．Methods Totally 30 female tree shrews （Tupaiabelangeri） were randomly divided into the treatment group， the model group， and the control group， with 10 in each group. Bilateral ovaries and uterus were ectomized for 6 months in the treatment group and the model group， and right fibula fractures were induced after confirmed osteoporosis. Tree shrews in the treatment group were administered with Osteoking by gastrogavage， once per two days. Equal volume of normal saline was given to tree shrews in the model group and control group. Blood biochemical indexes were detected at 3 months after fracture．Results Compared with those in the control group， serum creatine kinase isoenzyme （CK-MB）， alanine aminotransferase （ALT）， magnesium （Mg2 ）+ levels significantly increased （P＜0.01） while total cholesterol （CHOL） and blood glucose （Glu） levels were significantly decreased （P＜0.01） in the OPF model group. Compared with those in the model group， serum CK-MB and ALT significantly decreased （P＜0.01） and Mg2+ significantly decreased（P＜0.05 while CHOL and Glu significantly increased （P＜0.05） in the treatment group. Conclusion Osteoking can protect the heart and liver function of tree shrew with osteoporotic fracture. [ABSTRACT FROM AUTHOR]

Objective Osteoking，China Yunnan Yi，is a local Chinese herbal medicine．It has been used to treat osteoporotic fracture，but its therapeutic effect on osteoporosis is not clear．The purpose of this experiment is to study the effects of Osteoking on ovariectomized female rat osteoporosis model． Methods Thirty-six female SD rats were randomly divided into three groups (each group of 12): the control，the surgery and the dose groups． The dose and the surgery groups were operated with bilateral oophorectomy，and at the same time the dose group was fed with Osteoking． Bone mineral density and bone mineral salt content were measured at the first and 10th week． Results At 10 weeks the total BMD of the dose group was significantly higher than that of the surgery group (by 1. 84%，P ＜ 0. 05)． The total BMD of the dose group was 1. 2% lower than the control group，which did not reach statistical significance (P ＞ 0. 05) ． For total BMC: the dose ＞ the surgery ＞ the control． The body weight of the dose group was similar to that of the surgery group，with no significant difference (P ＞ 0. 05)．The body weight of both dose and surgery groups was significantly heavier than that of the control group (P ＜ 0. 05)．Conclusion Osteoking could reduce the loss of bone mass in ovariectomized female rat model of osteoporosis． [ABSTRACT FROM AUTHOR]

Heng-Gu-Gu-Shang-Yu-He-Ji, also known as OsteoKing, is used as a herbal Traditional Chinese Medicine for the treatment of bone disease, including femoral head necrosis and osteoarthritis. However, whether OsteoKing has anti-osteoporotic properties has remained to be elucidated. The purpose of the present study was therefore to investigate the effects of OsteoKing on ovariectomy-induced osteoporosis in rabbits. Female New Zealand white rabbits were randomly divided into an ovariectomized (OVX) group and a sham-surgery group. The rabbits in the OVX group were subjected to an ovariectomy, while the rabbits in the sham group were subjected to the removal of an area of fat near the two ovaries. Bone mineral density, mechanical properties, serum biochemical parameters and micro-architecture were examined at 150 days post-OVX to characterize the experimental animal model. Once the osteoporotic rabbit model had been established, the rabbits in the OVX group were divided into the following groups: Model group, nilestriol group and 300 and 600 mg/kg OsteoKing groups, containing 16 rabbits in each group. OsteoKing and nilestriol were administered orally. The bone mineral density, mechanical properties, serum biochemical parameters, histology and micro-architecture were examined using dual-energy X-ray absorptiometric analysis, mechanical assessments, enzyme-linked immunosorbent assays, histopathological evaluation and micro-computerized tomography examination following 60 days and 120 days of treatment, respectively. Treatment with OsteoKing led to an elevation in the bone mineral density of the vertebra and serum phosphorus levels, reduced serum concentrations of osteocalcin, procollagen type I N-terminal peptide, tartrate-resistant acid phosphatase 5b and cross-linked N-telopeptide of type I collagen, improved mechanical properties (maximum load, stiffness and energy absorption capacity), and micro-architecture of the lumbar vertebra in the OVX osteoporotic rabbit model following treatment for 120 days. In conclusion, it was demonstrated that OsteoKing is effective in the prevention of estrogen deficiency-associated bone loss and may be a promising drug for the treatment of post-menopausal osteoporosis. [ABSTRACT FROM AUTHOR]

Objective: To study the influence of Osteoking combined with Jiegu Qili Pian on healing effect of distal radius fracture, so as to To provide basis for clinical treatment of distal radius fractures. Methods: 120 cases of patients with distal radius fractures treated in our hospital from January 2014 to December 2015 were selected, the patients were divided into study group and control group according to the random number table method,60 cases in each group, they were given manipulative reduction and splint fixation, after which the study group was given Osteoking combined with Jiegu Qili Pian, while the control group was given Osteoking combined with placebo, course of treatment was 12 weeks in two groups. Evaluated the wrist function after treatment and the X-ray fracture healing scores in two groups 4th, 8th, 12th week after treatment, and compared the clinical healing time and adverse reactions in two groups after treatment. Results: The excellent and good rate of wrist function in the study group was 96.67%, which was significantly higher than 81.67％in the control group, the difference was statistically significant (P<0.05); The X-ray fracture healing scores in study group were significantly higher than control group 8th,12th week after treatment (P<0.05); The clinical healing time in study group was significantly shorter than control group(P<0.05), but there was no adverse reactions occurred in the two groups after treatment. Conclusion: Osteoking combined with Jiegu Qili Pian is conducive to promoting the healing of distal radius fracture, it is not only can improve the wrist function, but also safe and effective, which is worth to promote. [ABSTRACT FROM AUTHOR]

Introduction. Osteoporosis (OP) is characterized by microstructural degeneration of bone tissue, low bone mass, bone fragility and even brittle fracture (osteoporotic fracture, OPF). OP and OPF are common and there are many disadvantages to the current medications for OP/OPF. Osteoking is a traditional Chinese medicine (TCM) originating from the Yi nationality (Yunnan, China) that has been used to treat bone diseases for decades. Hypothesis/Gap Statement. This study will reveal the changes in the intestinal microbiota of OP rats after 70 days of osteoking treatment. Method. With duplication of sham and OP rats, eight groups were established, with six rats in each group. The intestinal microbiotas were analysed by 16S rDNA sequencing. Results. The results showed that osteoking changed the intestinal microbiota of sham rats and OP rats. The mechanism by which osteoking improves OP is related to the functions of the intestinal microbiota. After 70 days of treatment with osteoking, the contents of Pseudonocardia, Pedomicrobium, Variovorax, Niastella and Actinosynnema were decreased in OP rats. The functions of the above intestinal microbiota related to iron metabolism affected calcifediol and 25(OH)D, and measuring these bone metabolic indicators is required for further study. Conclusion. Osteoking changes the intestinal microbiota to improve OP, and further study which reveals these intestinal microbiota and mechanism is needed.

Objective: To evaluate the effect of Osteoking (...) in preventing postoperational deep venous thrombosis (DVT) in patients with intertrochanteric fracture (ITF). Methods: With prospective and randomized controlled clinical design adopted, 62 patients with ITF after operation were assigned into 2 groups, the tested group and the control group, Osteoking (25 ml every other day) and Sanchidansheng tablets (..., 3 tablets thrice a day) were given orally to them respectively for 10 days. Difference of round length of thighs and shanks between two sides were measured on the 10th day and Doppler ultrasonic examination on the fractured leg was carried out. Results: The occurrence rate of DVT in the tested goup was 9.4%, which was lower than that in the control group (30.0%, P<0.05). All the difference of round lengths, either that of the thigh or the shank, was less in the tested group than that in the control group, showing statistical significance (P<0.05). Conclusion: Osteoking has a satisfactory effect in preventing postoperational DVT in patients with ITF. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effectiveness of osteoking, a Traditional Chinese Medicine originating from Yi nationality, against osteoporosis (OP) and osteoporotic fracture (OPF), and to elucidate its mechanism of action., Methods: An osteoporotic fracture rat model was established; animals were divided into three treatment groups: parathyroid hormone, osteoking and 0.9%NaCl. After 4, 8 and 12 weeks of treatment, serum and bone tissues were collected. Enzyme-linked immuno sorbent assay, x-ray, histopathological evaluation and proteomics were used. Proteomics and GO annotation were performed based on identified peptides. The relative network was obtained from the STRING database and verified by polymerase chain reaction and Western blotting., Results: After osteoking treatment, the bone mineral density (BMD) increased with time in the osteoking group. At week 12, the BMD and bone mineral salt content of the osteoking group were 4.5% and 20.6% higher than those of the negative control group, respectively. Furthermore, the body weight followed the order of positive control group > osteoking group > negative control group, with significant differences among the groups (P < 0.05). Micro-CT analysis of femur sections revealed that the bone surface/volume ratio was significantly higher in the osteoking group than that in the negative control group. X-ray images demonstrated that the osteoking group showed clear callus. Moreover, high-voltage micro-CT demonstrated a massive cortical bone accumulation in the osteoking group. The gray values of callus in the osteoking group were higher than those in the negative group. From week 4 to 12, the serum bone alkaline phosphatase level increased by 49.6% in the osteoking group and the serum propeptide of type Ⅰprocollagen level decreased by 80.6%. Alizarin red staining demonstrated that the calcium deposition in the osteoking group was higher than that in the negative control group. Notably, the expression of Mgp, a key osteogenesis inhibitor, was lower in the osteoking group compared with the negative control group. Moreover, Sparc, bone morphogenetic protein-2 and Bglap expression was higher in the osteoking group through activation of the transforming growth factor-receptor activator of nuclear factor κB Ligand pathway., Conclusion: Osteoking treatment increased bone quality and promoted calcium deposition. The results suggest that osteoking inhibits Mgp through the TGF-β/RANKL pathway to improve OP/OPF.

Osteoking is a Traditional Chinese Medicine consisting of seven types of medicinal herbs originated from Yi nationality and has been used in clinic to treat bone diseases for thousands of years in China. Osteoking shows excellent clinical therapeutic effects on osteoporosis, but it is not clear whether Osteoking could exhibit beneficial effects against osteoporosis via reducing reactive oxygen species (ROS). Aim of the study: To explore whether the protective effects of Osteoking on osteoporosis related to ROS, we investigated the effects of Osteoking on osteogenesis differentiation under oxidative stress. The ovariectomized (OVX) osteoporosis model was established by ovarian surgery, and Osteoking was orally administrated for 84 days. Then the pathogenesis changes of femur were analyzed by Hematoxylin and eosin (H&E) and Masson's trichrome staining. The levels of ROS, malondialdehyde (MDA) and superoxide dismutase (SOD) from rats' serum were further measured. In vitro , mouse pre-osteoblastic MC3T3-E1 cells pre-treated with or without 0.25 mM tert -butyl hydroperoxide (t -BHP) for 2 h were cultured and treated with different dilutions of Osteoking or 20 μM N-Acetyl-L-cysteine for another 24 h, respectively. The intracellular ROS production and markers of oxidative damage of the MC3T3-E1 cells were determined using corresponding kits, respectively. The expressions of alkaline phosphatase (ALP), collagen type I, osteoprotegerin (OPG), TGF-β1, β-catenin, receptor activator of nuclear factor-κB ligand (RANKL) and interleukin-6 (IL-6) were further analyzed by qRT-PCR and western blotting upon treatment. Our results showed that Osteoking significantly improving trabecular microstructure by promoting collagen fiber repair and new bone or cartilage regeneration was demonstrated in OVX osteoporosis rat models by micro-CT analysis and histological staining results. Osteoking supplementation reduced the levels of ROS and MDA in OVX rat serum and increased SOD activities. In addition, Osteoking could also up-regulate the proteins expression levels of Runx2, osteocalcin (BGP) and osteoprotegerin (OPG) but reducing the expression of tartrate-resistant acid phosphatase (TRAP). In vitro , Osteoking could effectively inhibit the t -BHP-induced intracellular excessive ROS production and protect cells from oxidative stress in mouse pre-osteoblastic MC3T3-E1 cells. Meanwhile, the mRNA expressions of ALP, collagen type I, OPG, TGF-β1 and β-catenin were also up-regulated whereas the RANKL and IL-6 were down-regulated in Osteoking-treated MC3T3-E1 cells. A novel therapeutic mechanism of Osteoking on osteoporosis reveals by present investigation. Clinic effects of Osteoking to treat osteoporosis are closely related to its ability to reduce oxidative stress. Image 1 [ABSTRACT FROM AUTHOR]