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In this study, we isolated two new neoflavones from Aconitum austroyunnanense and examined their therapeutic activity against osteoarthritis of the knee. All the tested compounds displayed good anti-inflammatory activity and significantly therapeutic activity against osteoarthritis of rats using surgically induced rat OA models. Compound 1 has the best therapeutic activity and appears to be a promising candidate for further development.

OBJECTIVE: To investigate the effect of microencapsulated transgenic bone marrow mesenchymal stem cells (BMSCs) transplantation on early steroid induced osteonecrosis of femoral head (SONFH) in rabbits. METHODS: Alginate poly-L-lysine-sodium alginate (APA) microencapsulated transgenic BMSCs with high expression of Foxc2 were prepared by high-voltage electrostatic method. Part of the cells were cultured in osteoblasts and observed by alizarin red staining at 2 and 3 weeks. Forty New Zealand white rabbits were used to prepare SONFH models by using hormone and endotoxin. Thirty two rabbits who were successful modeling were screened out by MRI and randomly divided into 4 groups (groups A, B, C and D, n=8); another 6 normal rabbits were taken as normal control (group E). The rabbits in group A did not receive any treatment; and in groups B, C, and D were injected with normal saline, allogeneic BMSCs, and APA microencapsulated transgenic BMSCs respectively after core decompression. At 6 and 12 weeks after operation, specimens of femoral head were taken for HE staining to observe bone ingrowth; the expressions of osteocalcin (OCN), peroxisome proliferative activated receptor γ 2 (PPARγ-2), and vascular endothelial growth factor (VEGF) proteins were observed by immunohistochemistry staining. At 12 weeks after operation, the bone microstructure was observed by transmission electron microscope, and the maximum compressive strength and average elastic modulus of cancellous bone and subchondral bone were measured by biomechanics. RESULTS: After 2 and 3 weeks of induction culture, alizarin red staining showed the formation of calcium nodules, and the number of calcium nodules increased at 3 weeks when compared with 2 weeks. The rabbits in each group survived until the experiment was completed. Compared with groups A, B, and C, the trabeculae of group D were more orderly, the empty bone lacunae were less, there were abundant functional organelles, and obvious osteogenesis was observed, and the necrotic area was completely repaired at 12 weeks. Immunohistochemical staining showed that, at 6 and 12 weeks after operation, the expressions of OCN and VEGF in groups A, B, and C were significantly lower than those in groups D and E, while those in groups B and C were significantly higher than those in group A, and in group E than in group D (P0.05). CONCLUSION: In vivo transplantation of microencapsulated transgenic BMSCs can repair early SONFH in rabbits.

Background Mechanical stress has been well known to be a significant risk factor for the onset and progression of osteoarthritis (OA). The objective of this study is to clarify the key genes and the signaling pathways which is associated with the effect of mechanical stress on OA development. Results A total of 213 Differentially expressed genes (DEGs) were identified in cultured chondrocytes from knee and finger joints, including 101 up- and 112 down-regulated genes, which brought an enrichment in positive or negative regulation of cell proliferation, positive regulation of cell division, positive regulation of apoptotic process and embryonic limb morphogenesis. Two comparison cohorts shared 122 overlapping genes including 49 up- and 73 down-regulated genes, which enriched in DNA replication, cell division, calcium ion binding and positive regulation of apoptotic process. Twelve hub overlapped genes including Cyclin dependent kinase 1 (CDK1), Kinesin family member 11 (KIF11), Mitotic checkpoint serine/threonine kinase (BUB1) and Mitotic arrest deficient 2 like 1 (MAD2L1) were defined and predicted miRNAs of the genes mainly enriched in Transforming growth factor beta (TGF-β) signaling pathway, Wnt signaling pathway and Mitogen activated kinase-like protein (MAPK) signaling pathway. Conclusions Mechanical stress shown multiple influences on chondrocytes, physiologically necessary for maintaining chondrocyte homeostasis and promoting cartilage development, pathologically help for cartilage destruction and OA development via boosting chondrocyte differentiation and hypertrophy and accelerating cell apoptosis.

Ethnopharmacological relevance Gathering three ancient formulas, traditional Chinese medicine Bu-Shen-Tong-Luo decoction (BSTLD) has been used to treat postmenopausal osteoporosis (PMO) at the Jiangsu Province Hospital of Chinese Medicine for decades. However, the effect of BSTLD on angiogenesis and bone resorption as well as its possible mechanism are still unknown. Aim of the study This study was aimed to evaluate the preventive effect of BSTLD on ovariectomy-induced bone loss and vasculature disorder, and to investigate the possible bone protection mechanism of BSTLD in inhibiting bone resorption by enhancing angiogenesis signaling in ovariectomy-induced osteoporosis of rats. Materials and methods The animal experiment was divided into five groups. Rats underwent either sham surgery with intact ovaries (SHAM, n = 10) or bilateral ovariectomy (OVX, n = 40). OVX rats were randomly divided into four groups and gavaged by water (vehicle, 12 mL/kg, n = 10), BSTLD (6 g/kg, n = 10), BSTLD (12 g/kg, n = 10) and 17β-estradiol (E2, 100 μg/kg, n = 10) daily for 12 weeks, respectively. The bone loss and microstructure of the distal femur were observed using micro-computed tomography (μCT). The biomechanical parameters of the femur were detected using three-point bending tests. The distribution of osteoclasts and endothelial cells were analyzed by immunohistochemistry. The mRNA and protein levels of angiogenesis-related hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF), as well as osteoclast activation-related signaling calcitonin receptor (CALCR), cathepsin K (CTSK), receptor activator of NF-κB ligand (RANKL), osteoprotegerin (OPG), and β-catenin were assayed by RT-PCR or Western blot. Results BSTLD protected trabecular bone mass density and trabecular bone microstructure from ovariectomy-induced osteoporosis in rats. BSTLD significantly reduced mRNA and protein levels of calcitonin receptor and CTSK in femoral metaphysis and inhibited bone resorption in ovariectomized rats. Furthermore, BSTLD stabilized HIF-1α activity and subsequently increased VEGF expression to enhance angiogenesis and modulated RANKL/OPG signaling in this animal model. Conclusions These results demonstrated that BSTLD reduced osteoclasts activation and bone resorption in ovariectomy-induced osteoporosis. Bone protection by BSTLD may be associated with its stimulation of HIF-1α/VEGF angiogenesis signaling and suppression of RANKL/OPG ratio. This study may provide evidence that BSTLD treats postmenopausal osteoporosis, especially with micro-circulation complication.

In this paper, citrate-stabilized CuS nanocrystals were modified by NH 2 -terminated aptamer of carcinoembryonic antigen to fabricate aptamer-CuS complex via carbodiimide-activated coupling. Then, the complex was conjugated with graphene oxide (GO) to form aptamer-CuS/GO conjugates via π-π stacking interactions. Finally, glucosamine (Glu) was loaded into aptamer-CuS/GO conjugates to prepare aptamer-CuS/GO/Glu composites. The composites enabled CEA-targeted and pH-sensitive Glu release. Under near-infrared light irradiation at 980 nm, the composites had photothermal-accelerated release of Glu and chemo-photothermal synergistic therapy in vitro . Due to combined advantages from tumor biomarker-targeted, pH-sensitive, photothermal-accelerated drug release, as well as chemo- photothermal therapy, the composites could be developed towards multifunctional drug-delivery systems for highly efficient treatment against tumor cells.

The Chinese medicine compound, Jisuikang, can promote recovery of neurological function by inhibiting lipid peroxidation, scavenging oxygen free radicals, and effectively improving the local microenvironment after spinal cord injury. However, the mechanism remains unclear. Thus, we established a rat model of acute spinal cord injury using a modified version of Allen's method. Jisuikang (50, 25, and 12.5 g/kg/d) and prednisolone were administered 30 minutes after anesthesia. Basso, Beattie, and Bresnahan locomotor scale scores and the oblique board test showed improved motor function recovery in the prednisone group and moderate-dose Jisuikang group compared with the other groups at 3-7 days post-injury. The rats in the moderate-dose Jisuikang group recovered best at 14 days post-injury. Hematoxylin-eosin staining and transmission electron microscopy showed that the survival rate of neurons in treatment groups increased after 3-7 days of administration. Further, the structure of neurons and glial cells was more distinct, especially in prednisolone and moderate-dose Jisuikang groups. Western blot assay and immunohistochemistry showed that expression of brain-derived neurotrophic factor (BDNF) in injured segments was maintained at a high level after 7-14 days of treatment. In contrast, expression of nerve growth factor (NGF) was down-regulated at 7 days after spinal cord injury. Real-time fluorescence quantitative polymerase chain reaction showed that expression of BDNF and NGF mRNA was induced in injured segments by prednisolone and Jisuikang. At 3-7 days after injury, the effect of prednisolone was greater, while 14 days after injury, the effect of moderate-dose Jisuikang was greater. These results confirm that Jisuikang can upregulate BDNF and NGF expression for a prolonged period after spinal cord injury and promote repair of acute spinal cord injury, with its effect being similar to prednisolone.

MicroRNAs (miRs) have been reported to serve key roles in cancer. To investigate the function of miR-885-5p in osteosarcoma, the expression levels of miR-885-5p were analyzed in 85 osteosarcoma tissue samples and adjacent non-cancerous tissue samples, using reverse transcription-quantitative polymerase chain reaction analysis. It was demonstrated that miR-885-5p was downregulated in osteosarcoma tissues and cell lines. Notably, the expression level of miR-885-5p was closely associated with tumor size, Tumor-Node-Metastasis stage and lymph node metastasis. Additionally, low expression levels of miR-885-5p also predicted a poor prognosis of osteosarcoma. To further decipher the roles of miR-885-5p in osteosarcoma, it was determined that β-catenin, a key component of the Wnt signaling pathway, was a target of miR-885-5p. Furthermore, several functional experiments, including a colony formation assay, CCK-8 assay, wound healing assay and Transwell invasion assay, revealed that miR-885-5p suppressed cell proliferation, migration and invasion through inhibition of β-catenin. The results of the present study provide a novel insight into the molecular roles of miR-885-5p in osteosarcoma.

To investigate the role of the forkhead/Fox transcription factor 2 (Foxc2) over-expression in regulating osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by Wnt-β-catenin signaling pathwaysThe recombinant lentivirus carrying green fluorescent protein (group A) or Foxc2 (group B) were used to transfect the fifth generation rabbit BMSCs, and untransfected BMSCs served as a control (group C). The cell viability was measured with water soluble tetrazolium-1 (WST-1) regent at 72 hours after transfection. After 2 weeks of transfection, the expression of β-catenin in BMSCs was detected by real time fluorescence quantitative PCR, Western blot, and immunofluorescence staining. Meanwhile, the β-catenin inhibitors XAV-939 (0, 0.1, and 1.0 μmol/L) was added in group B; at 2 weeks after osteogenic and adipogenic induction, the gene and protein expressions of collagen type I (COL I), osteocalcin (OCN), and peroxisome proliferator activated receptor gamma 2 (PPARγ-2) were detected by real time PCR and Western blot.WST-1 results showed that the cell viability of group B (130.85%±0.15%) was significantly higher than that of group A (100.45%±0.35%) (The over-expression of Foxc2 gene in BMSCs may promote osteogenic differentiation by Wnt-β-catenin signaling pathway.观察过表达插头框转录因子C2（forkhead/Fox transcription factor 2，Foxc2）经Wnt-β链蛋白（β-catenin）信号通路调节兔BMSCs成骨分化，为基因转染BMSCs修复股骨头坏死提供理论依据。.利用慢病毒携带Foxc2或绿色荧光蛋白（green fluorescent protein，GFP）基因的重组慢病毒载体Lv-GFP（A组）及Lv-Foxc2（B组）转染第5代兔BMSCs，以未转染BMSCs为对照（C组）。慢病毒转染后72 h采用水溶性四氮唑-1（water soluble tetrazolium-1，WST-1）法检测细胞活性；慢病毒转染2周，通过免疫荧光染色、Western blot及实时荧光定量PCR检测过表达Foxc2对β-catenin表达水平的影响。然后，在B组中添加不同剂量（0、0.1、1.0 μmol/L）β-catenin抑制剂XAV-939，成骨、成脂诱导2周后，采用Western blot和实时荧光定量PCR检测各组成骨因子Ⅰ型胶原（collagen typeⅠ，COLⅠ）、骨钙素（osteocalcin，OCN）及成脂因子过氧化物酶体增殖体激活受体γ2（peroxisome proliferator activated receptor gamma 2，PPARγ-2）蛋白和基因的表达。.WST-1检测示，转染72 h B组细胞活性为130.85%±0.15%，显著高于A组的100.45%±0.35%，差异有统计学意义（过表达Foxc2通过调节Wnt-β-catenin信号通路促进BMSCs成骨分化。.

Age is a major risk factor for developing chronic diseases, including type 2 diabetes and osteoporosis. Emerging evidences suggest that the disorder of bone metabolism in osteoporosis is involved in the pathogenesis of glucose intolerance, insulin resistance and type 2 diabetes. However, their etiology and relative regulatory factors still remain elusive to clinicians and researchers. In this study, rats were divided into two groups: normal sham surgery control and ovariectomized (OVX) groups. We evaluated the global bone parameters, glucose metabolism, protein and gene expressions in both skeletal muscle and adipocytes. The present findings showed that the bone mineral density (BMD) and compression load of bone were markedly reduced in OVX rats as revealed by micro-CT, dual energy X-ray absorptiometry and bone biomechanics analysis. Besides, plasma estrogen, total alkaline phosphatase (TALP) and osteocalcin levels were significantly decreased in the OVX rats, but body weight, fat mass and plasma tartrate-resistant acid phosphatase (TRAP) and chemerin levels were significantly increased in the OVX rats. More interestingly, we found that p-AKT, p-P38MAPK, glucose transporter 4 (GLUT4) and peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) contents as well as GLUT4 and PGC-1α mRNA expression were significantly decreased in skeletal muscle and adipocytes of OVX rats. In conclusion, our results indicated that whole-body glucose metabolism and glucose intolerance in OVX rats was degressive, suggesting there was a novel link between osteoporosis and whole body glucose homeostasis, which are controlled by the P38MAPK/PGC-1α/GLUT4 signaling pathway.

Background: Osteoporosis is a disease of bones that leads to an increased risk of fracture. Epimedii Folium is commonly used for treating bone fractures and joint diseases for thousands of years in China. Methods: This study was aimed to screen active components, which might have the potency to stimulate osteoblasts proliferation and differentiation in Epimedii Folium. An HPLC method was established to analyze the main components in Epimedii Folium. The MTT and ALP methods were utilized for the assay of osteoblasts proliferation and differentiation activity. Bavachin, a flavonoid compound was treated as the positive control. Results: Totally eight compounds have been identified by comparing their retention time with correspondent standard substances. Icariside I and icariside II significantly stimulated cell proliferation and osteoblasts differentiation. All these compounds were found with a characterized flavonoid structure in each of their molecule backbones. Conclusion: These results lead to a hypothesis that flavonoid monoglycoside structure might be crucial to exhibit the activity. The structure–effect relationship of these compounds with flavonoid monoglycoside structure in mouse primary calvarial osteoblasts needs to be explored in further research. SUMMARY Eight compounds were identified by comparing their retention time with correspondent standard substances.Icariside I and icariside II significantly stimulated cell proliferation and osteoblasts differentiation.Flavonoid monoglycoside structure might be crucial to exhibit the osteoblasts proliferation and differentiation activity. Effects of the main components of Epimedii Folium on osteoblasts proliferation after treating 48 h. Abbreviations used: HPLC: High performance liquid chromatography, MTT: Methylthiazolyldiphenyl - tetrazolium bromide, ALP: Alkaline phosphatase

Osteoarthritis (OA) is a complex disease characterized by cartilage degeneration, secondary synovial membrane inflammation and subchondral bone changes. In recent years, many studies have confirmed that interleukin-18 (IL-18) is involved in the inflammatory process of inflammatory joint diseases. In the present study, we investigated IL-18 levels in plasma, synovial fluid and articular cartilage of patients with primary knee OA (n = 33) to analyze their relationship with radiographic severity. Compared to healthy controls (n = 15), OA patients had higher plasma and synovial fluid IL-18 concentrations (45.8 ± 22.1 vs. 23.7 ± 13.6 pg/ml, P

Developing efficient solutions to achieve automatic service composition has drawn significant attentions in service computing. As a composite service typically runs in a dynamic environment, adaptability of the composition solution arises as a central concern. Reinforcement Learning (RL) is one commonly used approach in service composition to achieve self-adaptability. However, traditional RL methods cannot guarantee good efficiency for large-scale composition problems. Hierarchical RL (HRL) has appeared to be a viable solution to address the efficiency issue. The applicability of existing HRL methods (e.g., MAXQ) requires a task graph, which can be generated by decomposing a composition plan into a task hierarchy. Current approaches that apply HRL to service composition generate the task graph manually, which will not scale to large service composition problems. In this paper, we address the above issue by systematically integrating automatic task decomposition and MAXQ HRL, resulting in an adaptive composition solution with good efficiency. Our experimental results demonstrate the effectiveness of the proposed service composition approach.

To develop an intelligent adjustable leg raising system which could realize the quantification and visualization of lower limbs' elevation angle and frequency.We determined the pole adjustable length of thighs and cruses and the flexion angle range of hips and knees according to the requirements of clinical lower limb function rehabilitation, and made force analysis of hip's and knee's flexion motivation mechanism and clinical observation on its security and effectiveness.This device was small and compact in overall appearance, which could adjust the angle of the hips and knees bending and ankles turning alone. The force analysis of the hips and knees flexion power element was consistent with the design requirements. The preliminary studies showed the device could relieve pain, improve the range of motion and promote the rehabilitation, which was superior to that of the Brown Ska (P0.05).The intelligent adjustable leg raising system meets the requirements of clinical usage, which is suitable for different heights, and the flexion angle ranges of hips and knees are wide and in high accuracy, which is worth of being improved and generalized.