Duchenne muscular dystrophy: RANK/RANKL/OPG (receptor activator of nuclear factor-kB/RANK ligand/osteoprotegerin) system and glucocorticoids / Distrofia muscular de Duchenne: el sistema RANK/RANKL/OPG (receptor activador del factor nuclear-kB/ligando de RANK/osteoprotegerina) y glucocorticoides

Abstract Duchenne muscular dystrophy (DMD) is an X-linked inherited disorder. Patients present with decreased bone mineral density (BMD) due to glucocorticoid therapy and progressive muscle weakness. Bone remodeling allows bone volume and structure to be maintained and controlled by local and systemic factors. These include the receptor activator of the nuclear factor-kB (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system, a determining pathway in the balance between bone formation and resorption. Disruptions in this complex, caused by factors such as glucocorticoids, can affect bone metabolism. The extensive action of the RANK/RANKL/OPG pathway suggests an influence on dystrophic muscle pathophysiology. This review aimed to highlight some aspects of the RANK/RANKL/OPG system, the effect of glucocorticoids on this pathway, and the pathophysiology of the patient with DMD.

Resumen La distrofia muscular de Duchenne (DMD) es un trastorno hereditario ligado al cromosoma X. Los pacientes presentan una disminución de la densidad mineral ósea (DMO) debido a los efectos adversos del tratamiento con glucocorticoides y a la debilidad muscular progresiva. El remodelado óseo permite mantener el volumen y la estructura ósea, proceso controlado por factores locales y sistémicos. Entre ellos destaca el sistema del receptor activador del factor nuclear-kB (RANK), su ligando natural RANKL (RANKL) y la osteoprotegerina (OPG), una vía determinante en el equilibrio entre la resorción y formación ósea. Las alteraciones en este complejo, originadas por factores como los glucocorticoides, pueden afectar el metabolismo óseo. La amplia acción de RANKL y OPG ha sugerido una influencia en la fisiopatología de la DMD. El objetivo de esta revisión fue destacar algunos aspectos del sistema RANK/RANKL/OPG, el efecto de los glucocorticoides en esta vía y la fisiopatología del paciente con DMD.

Effect of enhanced masticatory force on OPG, RANKL and MGF in alveolar bone of ovariectomized rats

Abstract Menopause induces oral bone loss, leading to various oral diseases. Mastication importantly affects bone metabolism in the jawbone. Objective: To analyze the effect of enhanced masticatory force on osteoprotegerin (OPG), receptor activator of nuclear factor kappa B ligand (RANKL), and mechano-growth factor (MGF) in alveolar bone of ovariectomized rats and to study the mechanics mechanism of the alveolar bone of ovariectomized rats response to enhanced masticatory force. Methodology: Thirty Sprague Dawley rats were randomly divided into three groups: sham-operation group (fat around the removed ovary + normal hard diet), model group (ovariectomy + normal hard diet), and experimental group (ovariectomy + high hard diet). It was a 2-month experiment. Enzyme-linked immunosorbent assay (ELISA) detected serum estradiol (E2), osteocalcin (BGP) and alkaline phosphatase (ALP) in rats. Bone histomorphometric indices in the third molar region of maxilla were detected by micro-CT; protein expressions of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Western blot; and gene expression of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Quantitative Real-Time PCR. Results: Comparing with model group, serum E2 in experimental group increased but not significantly, serum BGP and serum ALP in experimental group decreased but not significantly, OPG in experimental group in alveolar bone increased significantly, RANKL in experimental group in alveolar bone decreased significantly, RANKL/OPG ratio in experimental group decreased significantly, MGF in experimental group in alveolar bone increased significantly, bone volume to total volume fraction increased significantly in experimental group, trabecular thickness increased significantly in experimental group, and trabecular separation decreased significantly in experimental group. Conclusion: Enhanced masticatory force affected the expression of OPG, RANKL, and MGF in alveolar bone of ovariectomized rats, improved the quality of jaw bone of ovariectomized rats, and delayed oral bone loss by ovariectomy.

Effect of Decomposed Zuoguiwan Recipes in Improving BMD of Ovariectomized Rats by Regulating OPG/RANKL Pathway Mediated by β2AR / 中国实验方剂学杂志

Objective:To investigate the mechanism of decomposed Zuoguiwan（ZGW） recipes in treating ovariectomized osteoporosis rats. Method:Forty Sprague-Dawley female rats were equally and randomly divided into Sham-operated group, ovariectomized model group, positive group, and low and high-dose ZGW groups. After 12 weeks of administration by gavage, the bone mineral density (BMD) of rats' distal femur was measured by micro-CT, the morphology of bone tissue were observed by hematoxylin-eosin staining (HE), β-cross-linked c-telopeptide of type Ι collagen (β-CTX) and bone-specific alkaline phosphatase (BALP) in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the mRNA and protein expressions of β2AR, OPG and RANKL were evaluated by Western blot analysis and real-time quantitative polymerase chain reaction （PCR）. Result:Compared with Sham-operated group, BMD of rats in ovariectomized model group was decreased (P<0.01), morphology of bone tissue was destroyed, serum BALP was reduced, while β-CTX was boosted (P<0.01)，mRNA and protein expressions of OPG in tibia were reduced, while RANKL were increased, and mRNA and protein expressions of β2AR in the hypothalamus were decreased (P<0.05, P<0.01). Compared with ovariectomized model group, BMDs of rats in low and high-dose ZGW groups were increased (P<0.01), morphology of bone tissue was repaired, serum BALP and mRNA and protein expressions of OPG in tibia were up-regulated (P<0.05, P<0.01), whereas serum β-CTX and mRNA and protein expressions of β2AR in the hypothalamus and RANKL in tibia were down-regulated (P<0.05, P<0.01). Conclusion:Yang-nourishing components in decomposed Zuoguiwan recipes can improve BMD of ovariectomized rats by regulating OPG/RANKL pathway mediated by β2AR. "Seeking Yin in Yang" is a crucial mechanism of Zuoguiwan in treating ovariectomized osteoporosis in rats.

East java extract propolis as potential intracanal medicament in experimentally induced chronic apical periodontitis

Introduction: A persistent infection after cleaning and shaping root canal is the main etiology of root canal treatment failure. Enterococcus faecalis has been considered as one of the most resistant species in root canal treatment. E. faecalis can stimulate receptor activator of nuclear factor-kappa B ligand (RANKL) which can increase nuclear factor of activated T-cell (NFATc1) in chronic apical periodontitis. East Java propolis has antibacterial effects and is biocompatible with in vitro effects. Aim: This study is aimed to analyze the East Java propolis extract as potential intracanal medicament in chronic apical periodontitis caused by E. faecalis bacterial infection. Materials and Methods: This study used 30 Wistar rats divided into three groups. In Group I, the first upper right molar tooth as healthy tooth was used for negative control group. In Group II, the first upper right molar tooth was used for a prepared root canal, and 10 ml brain heart infusion broth containing E. faecalis ATCC29212 106 CFU was injected into the canal and restored with glass-ionomer cement (GIC) for the experimentally induced chronic apical periodontitis group. In Group III, after root canal preparation, E. faecalis ATCC 29212 106 CFU was injected, and then, 10 μl propolis applied and tooth restored with GIC. It took 21 days for the periapical lesions to develop after pulp infection. The rats were then sacrificed to conduct immunohistochemical examinations in order to measure the expressions of RANKL and NFATc1. Results: The average of RANKL and NFATc1 expression in Group III was significantly lower than those in the experimentally induced chronic apical periodontitis group (P < 0.05). Conclusion: It can be concluded that East Java propolis extract is a potential intracanal medicament through the study of experimentally induced chronic apical periodontitis caused by E. faecalis infection in Wistar rats.

Effect of Vitamin D during Orthodontic Tooth Movement on Receptor Activator of Nuclear Factor Kappa-Β Ligand Expression and Osteoclast Number in Pregnant Wistar Rat (Rattus novergicus)

Background: Female patients have the possibility tobecome pregnant during orthodontic treatment. VitaminD usually consumed by pregnant women. Estrogen andVitamin D could affect bone metabolism. Aim andObjectives: The aim of this study was to analyze theeffect of vitamin D during orthodontic movement inpregnant rats by Receptor Activator of Nuclear FactorKappa-Β Ligand (RANKL) expression and osteoclastnumber. Material and Methods: The experimentalobservational analytic study with post-test only controlgroup design and simple random sampling method wasconducted. 24-healthy-female Wistar rats were dividedinto 4 groups; K1: pregnant rats with orthodontic toothmovement and vitamin D on Day 7; K2: pregnant ratswith orthodontic tooth movement and vitamin D on Day14; K3: pregnant rats with orthodontic tooth movementwithout vitamin D on Day 7 and; K4: pregnant rats withorthodontic tooth movement without vitamin D on Day214. Nickle-Titanium coil spring with 10 g/mm forcewas placed between the incisors and the maxillarymolars. The RANKL expression and osteoclastsnumber were analyzed using Analysis of Variance(ANOVA) (p<0.05). Results: The highest osteoclastsnumber (8.494 ± 1.194), and RANKLexpression (7.967± 2.185) found in K1 group with significant betweengroups (p<0.05).Conclusions: Vitamin D increaseosteoclast number and RANKL expression duringorthodontic tooth movement in pregnant rats.

The function of OPG/RANKL/RANK system in tumor bone metastasis / 中华检验医学杂志

Tumor bone metastasis is one of the main causes of treatment failure in tumor.Since tumor bone metastasis is a complex process,the exact etiopathogenesis of tumor bone metastasis has not been clarified.Recent studies demonstrated that osteoprotegerin/receptor activator of NF-κB ligand/receptor activator of NF-κB (OPG/RANKL/RANK) system plays an important role during the development and progression of tumor bone metastasis.In this review,the research progress of OPG/RANKL/RANK system,including biological functions,the bone microenvironment regulation,the detection method,the diagnosis and treatment of tumor will be summerized.

Effect of Zuoguiwan in Improving BMD and Trabecular Bone Microarchitecture in Ovariectomy-induced Osteoporosis Rats by Regulating RANKL/OPG Pathway Mediated by β2AR / 中国实验方剂学杂志

Objective: To investigate the mechanism of Zuoguiwan in treating ovariectomy-induced osteoporosis rats by receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) signaling pathway mediated by β2-adrenergic receptor (β2AR). Method: Forty Sprague-Dawley female rats were randomly divided into Sham-operated group (Sham) and four ovariectomized (OVX) subgroups. Rats in Sham and OVX groups were treated with 17β-estradiol (50 μg·kg-1·d-1), and low and high-dose ZGW (2.3,4.6 g·kg-1 lyophilized powder) for 3 months, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to measure the serum markers of bone turnover. Micro-CT was used to evaluate and measure trabecular bone microarchitecture and bone mineral density (BMD) of the right distal femur. Western blot analysis and Real-time PCR were used to measure mRNA and protein expressions of β2AR, OPG and RANKL. Result: After 12 weeks of treatment with Zuoguiwan, the level of serum β-cross-linked c-telopeptide of type Ι collagen (β-CTX) (PPPβ2AR in the hypothalamus (PPConclusion: The mechanism of Zuoguiwan in alleviating BMD and trabecular bone microarchitecture in ovariectomy-induced osteoporosis rats might be related to the regulation of RANKL/OPG Pathway mediated by β2AR.

Efficacy of Wenyangbushen Formula on mRNA Expression of Osteoprotegerin, Receptor Activator of Nuclear Factor-kappa B and Its Ligand in Rabbits with Steroid-induced Avascular Necrosis of Femoral Head / 中国康复理论与实践

Objective:To observe the effect of Wenyangbushen formula on mRNA expression of osteoprotegerin (OPG), receptor activator of nuclear factor- kappa B (RANK) and receptor activator of nuclear factor- kappa B ligand (RANKL) in rabbits with steroid-induced avascular necrosis of femoral head (SANFH). Methods:A total of 46 healthy conventional New Zealand white rabbits were randomly divided into normal group (n = 10) and model building group (n = 36). The modified method of horse serum plus methylprednisolone was used to establish the SANFH model. Two rabbits from the normal group and four from the model building group were used for HE staining. Then the other models were randomly divided into model group, and low-dose, medium-dose and high-dose treatment groups, with eight rabbits in each group. The normal group was given normal saline 10 ml/d, and the treatment groups were given Wenyangbushen formula 6.44 g/(kg·d), 9.66 g/(kg·d) and 12.88 g/(kg·d), respectively, for eight weeks. The mRNA expression of OPG, RANK and RANKL was detected by reverse transcription- polymerase chain reaction. Results:The empty lacuna rate was significantly higher in the model group than in the normal group (t = 17.085, P < 0.001). Compared with the model group, the mRNA expression of OPG increased (P < 0.01), and the mRNA expression of RANK and RANKL decreased (P < 0.01), in the treatment groups. Compared with the low-dose treatment group, the mRNA expression of OPG increased (P < 0.01), and the mRNA expression of RANK and RANKL decreased (P < 0.01), in the medium-dose and high-dose treatment groups. There was no significant difference in the mRNA expression of OPG, RANK and RANKL between the low-dose treatment group and the high-dose treatment group (P > 0.05). Conclusion:Wenyangbushen formula could increase the mRNA expression of OPG and inhibit the mRNA expression of RANK and RANKL in the femoral head tissue of the rabbits with SANFH.

Effect of salmon calcitonin on RANK/RANKL/OPG osteolysis pathway in human macrophages after particle induction / 中国医师杂志

Objective To explore the effect of salmon calcitonin on the receptor activator of NF-κB/receptor activator of NF-κB ligand/osteoprotegerin (RANK/RANKL/OPG) osteolysis pathway in human macrophages after particles induction.Methods The polyethylene wear particles were extracted from the periacetabular boundary membrane tissue of a patient with hip prosthesis loosening.The optimal reaction cell concentration of human macrophages to polyethylene wear particles (with a concentration of 0.1 mg/ml) was measured by methyl thiazolyl tetrazolium (MTT) assay.Particles were used to stimulate human macrophages,while salmon calcitonin with different drug concentrations was used for intervention.They were randomly divided into five groups,with six parts in each group:Group A,control group;Group B,particle group;Group C,particle + salmon calcitonin (10-8 mol/L) group;Group D,particle + salmon calcitonin (10-10 mol/L) group;Group E,particle+salmon calcitonin (10-12 mol/L) group.After cocultured for 48 hours,quantitative polymerase chain reaction (qPCR) assay was used to detect the mRNA expression of RANK,RANKL and OPG in macrophages.Results After induction with particles,Group B had a higher expression in RANK and RANKL,and lower OPG expression than Group A.After salmon calcitonin intervention,the expression of RANK and RANKL were significantly decreased,and OPG expression was significantly increased.Group B had the highest RANKL/OPG rate.After drug intervention,the RANKL/OPG rate in C,D,E group were reduced.Conclusions The particles can induce the transformation of macrophages into osteoclasts.In addition to directly inhibiting osteoclast activity,salmon calcitonin prevents macrophage from differentiating into osteoclasts through modulating RANKL/RANK/OPG signaling pathway.

Effect of caspases and RANKL induced by heavy force in orthodontic root resorption / 대한치과교정학회지

OBJECTIVE: Orthodontic root resorption (ORR) due to orthodontic tooth movement is a difficult treatment-related adverse event. Caspases are important effector molecules for apoptosis. At present, little is known about the mechanisms underlying ORR and apoptosis in the cementum. The aim of the present in vivo study was to investigate the expression of tartrate-resistant acid phosphatase (TRAP), caspase 3, caspase 8, and receptor activator of nuclear factor kappa-B ligand (RANKL) in the cementum in response to a heavy or an optimum orthodontic force. METHODS: The maxillary molars of male Wistar rats were subjected to an orthodontic force of 10 g or 50 g using a closed coil spring. The rats were sacrificed each experimental period on days 1, 3, 5, and 7 after orthodontic force application. And the rats were subjected to histopathological and immunohistochemical analyses. RESULTS: On day 7 for the 50-g group, hematoxylin and eosin staining revealed numerous root resorption lacunae with odontoclasts on the root, while immunohistochemistry showed increased TRAP- and RANKL-positive cells. Caspase 3- and caspase 8-positive cells were increased on the cementum surfaces in the 50-g group on days 3 and 5. Moreover, the number of caspase 3- and caspase 8-positive cells and RANKL-positive cells was significantly higher in the 50-g group than in the 10-g group. CONCLUSIONS: In our rat model, ORR occurred after apoptosis was induced in the cementum by a heavy orthodontic force. These findings suggest that apoptosis of cementoblasts is involved in ORR.

Bone-targeted therapies to reduce skeletal morbidity in prostate cancer / 亚洲男科学杂志(英文版)

Bone metastases are the main driver of morbidity and mortality in advanced prostate cancer. Targeting the bone microenvironment, a key player in the pathogenesis of bone metastasis, has become one of the mainstays of therapy in men with advanced prostate cancer. This review will evaluate the data supporting the use of bone-targeted therapy, including (1) bisphosphonates such as zoledronic acid, which directly target osteoclasts, (2) denosumab, a receptor activator of nuclear factor-kappa B (RANK) ligand inhibitor, which targets a key component of bone stromal interaction, and (3) radium-223, an alpha-emitting calcium mimetic, which hones to the metabolically active areas of osteoblastic metastasis and induces double-strand breaks in the DNA. Denosumab has shown enhanced delay in skeletal-related events compared to zoledronic acid in patients with metastatic castration-resistant prostate cancer (mCRPC). Data are mixed with regard to pain control as a primary measure of efficacy. New data call into question dosing frequency, with quarterly dosing strategy potentially achieving similar effect compared to monthly dosing for zoledronic acid. In the case of radium-223, there are data for both pain palliation and improved overall survival in mCRPC. Further studies are needed to optimize timing and combination strategies for bone-targeted therapies. Ongoing studies will explore the impact of combining bone-targeted therapy with investigational therapeutic agents such as immunotherapy, for advanced prostate cancer. Future studies should strive to develop biomarkers of response, in order to improve efficacy and cost-effectiveness of these agents.

Expression of osteoprotegerin and receptor activator of nuclear factor kappa-B ligand in mandibular ramus osteotomy healing with administration of different doses of parathyroid hormone / 中华口腔医学杂志

Objective@#To investigate the effect of parathyroid hormone (PTH) on the bone healing of mandibular ramus osteotomy.@*Methods@#The mandibular ramus osteotomy model was established in sixty rabbits and these rabbits were randomly divided into experimental group A, experimental group B and control group. In the experimental group A and experimental group B, the rabbits were given PTH (20 and 40 μg/kg respectively) every other day after operation. In the control group, 1 ml saline was given. The animals were sacrificed at 1 week, 2 weeks, 3 weeks and 4 weeks postoperatively. The new bone formation was observed by histology and cone bone CT. The expression of osteoprotegerin and receptor activator of nuclear factor kappa-B (RANKL) in the new bone was detected by real-time quantitative PCR.@*Results@#The experimental groups has better osteogenesis and the bone mineral density than the control group in osteotomy area. The experimental group B showed the best osteogenesis.Osteoprotegerin mRNA expression in experimental group A (1.127±0.035, 1.742±0.049, 1.049±0.062, 1.063±0.036) was significantly higher than that in the control group in each period (0.965±0.082, 1.254±0.071, 0.793±0.061, 0.684±0.055) (P=0.010, P=0.000, P=0.001, P=0.020), while group B (1.416±0.205, 2.648±0.168, 1.652±0.091, 1.712±0.070) was significantly higher than group A (P=0.000, P=0.010, P=0.023, P=0.003). RANKL mRNA expression in control group (1.666±0.086, 1.058±0.105, 0.885±0.124, 0.972±0.136) was significantly higher than that of the group A (0.788±0.036, 0.585±0.017, 0.692±0.017, 0.527±0.051) (P=0.001, P=0.006, P=0.003, P=0.028) in each period, while group A was significantly higher than group B(0.247±0.022, 0.240±0.034, 0.134±0.011, 0.103±0.050) (P=0.000, P=0.001, P=0.002, P=0.012).@*Conclusions@#PTH can upregulate the expression of osteoprotegerin and reduce expression of RANKL, thus promoting new bone formation. Intermittent administration of high dose of parathyroid hormone can further promote the healing process after mandibular ramus osteotomy.

Role of osteoprotegerin and receptor activator of nuclear factor-κB ligand in bone toxicity in rats co-exposed to fluoride and arsenite / 中华地方病学杂志

Objective To analyze the role of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) in bone toxicity in rats co-exposed to fluoride and arsenite.Methods One hundred and ninety-two 8-week-old clean-grade Wistar rats weighing (200 ± 50) g were divided into 16 groups by weight using random number table method of 12 rats in each group by 2 × 4 factorial experimental design (half female and half male),and treated with different doses of fluoride,arsenite and fluoride plus arsenite in deionized water (untreated control group containing 0.0 mg/kg fluoride and 0.0 mg/kg arsenite;low-,moderate-,and high-fluoride groups supplemented with 5.0,10.0 and 20.0 mg/kg fluoride and 2.5,5.0 and 10.0 mg/kg arsenite) for 6 months.Rats were divided into control (F0As0),low fluorine (F5.0As0),moderate fluoride (F10.0As0),high fluoride (F20.0As0),low arsenic (F0As2.5),moderate arsenic (F0As5.0),high arsenic (F0As10.0),low fluorine and low arsenic (F5.0As2.5),low fluorine and moderate arsenic (F5.0As5.0),low fluorine and high arsenic (F5.0As10.0),moderate fluorine and low arsenic (F10.0As2.5),moderate fluorine and moderate arsenic (F10.0As5.0),moderate fluorine and high arsenic (F10.0As10.0),high fluorine and low arsenic (F20.0As2.5),high fluorine and moderate arsenic (F20.0As5.0),high fluorine and high arsenic (F20.0As10.0) groups.The protein expressions of OPG and RANKL in bone were measured via the enzyme-linked immunosorbent assay method.The mRNA expressions of OPG and RANKL were measured with quantitative real-time PCR.Results Compared with F0As0 [(2.678 ± 0.136) ng/mg,(29.658 ± 0.662) pg/mg],the protein expressions of OPG [(2.857 ± 0.162),(2.983 ± 0.272),(3.117 ± 0.143) ng/mg],and RANKL [(32.533 ± 0.999),(32.698 ± 1.932),(33.331 ± 1.140) pg/mg] in F5.0As0,F10.0As0,F20.0As0 were increased with increasing of fluoride doses;increased first and then decreased was observed in levels of RANKL protein [(32.348 ± 2.838),(31.589 ±1.359),(28.843 ± 1.908) pg/mg] in F0As2.5,F0As5.0,F0As10.0 with increasing of arsenic doses (P＜0.05).Compared with F0As0 (0.83 ± 0.19,0.92 ± 0.23),the mRNA expressions of OPG (1.14 ± 0.27,1.33 ± 0.39,1.69 ± 0.77) and RANKL (1.02 ± 0.21,1.17 ± 0.15,1.25 ± 0.31) in F5.0As0,F10.0As0,F20.0As0 were increased with increasing of fluoride dose.Fluoride had a significant effect on protein and mRNA expressions of OPG and RANKL (F=11.530,21.765,6.320,3.543,P ＜ 0.05).There was interaction between fluoride and arsenite on the expressions of RANKL protein and mRNA,OPG protein (F =9.496,2.217,3.375,P ＜ 0.05).Conclusion When rat is co-exposed to fluorine and arsenic,fluorine plays a leading role in regulating RANKL and OPG,and arsenic is indirectly involved in the fluorine bone toxicity in rats,fluorine and arsenic has a antagonistic effect on OPG and RANKL expressions.

The effects of different concentrations of crocin on receptor activator of nuclear factor kappa B ligand(RANKL)–induced osteoclastogenesis / 天津医药

Objective To investigate the effects of different concentrations of crocin on receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis using the monocyte-macrophage cell line RAW264.7. Methods The monocyte-macrophage cell line RAW264.7 was cultured routinely.After treatment with 0,6.25,12.5,25, 50,100,200 and 400 μmol/L crocin,cell counting kit-8(CCK-8)assay was used to analyze the viability of RAW264.7 cells to screen out the appropriate experimental concentration. RAW264.7 cells were induced by RANKL (100 ng/L) to form osteoclasts. After treated with 0, 12.5, 50 and 100 μmol/L crocin respectively, the number of osteolasts was counted by tatrate resistant acid phosphatasec (TRAP) staining. Real-time PCR was used to detect the mRNA expression levels of calcitonin receptor(CTR),nuclear factor of active T cells 1(NFATC1),C-fos and TRAP.Results No significant effects of crocin (within 0-100 μmol/L) were found on the viability of RAW264.7 cells (P>0.05). However, When crocin concentration was over 100 μmol/L,the cell proliferation was decreased,and which showed a significant inhibitory effect on proliferation (P<0.05). Thus, 0-100 μmol/L crocin was selected as the experiment concentration. The amount of differentiated osteolasts and the expression levels of CTR,NFATC1,C-fos and TRAP mRNA were decreased significantly with the increased concentrations of crocin(P<0.05).Conclusion At a certain concentration(0-100 μmol/L),the higher levels of crocin could inhibit RANKL-induced osteoclastogenesis.

Efficacy and safety of denosumab combined with surgery in treatment of giant cell tumor of pelvis / 肿瘤研究与临床

Objective To evaluate the safety and efficacy of denosumab in treatment of patients with pelvic giant cell tumor of bone (GCTB) during perioperative period. Methods This is a retrospective observational study. Twenty-three patients diagnosed with pelvic GCTB undergoing perioperative denosumab treatment in Musculoskeletal Tumor Center of Peking University People's Hospital from January 2014 to December 2016 were reviewed. The subjective adverse reactions and mandibular X-ray films were used to assess the drug safety. As for efficacy, imaging findings (including X-ray, CT, magnetic resonance imaging) were reviewed. MSTS-93 scoring system was applied in the postoperative functional assessment. Histological response rate, objective response rate, clinical benefit rate and event-free survival rate were all used to deficit the efficacy of denosumab in the treatment of pelvic GCTB combined with surgery. All the results of postoperative were compared statistically with pelvic GCTB patients who underwent surgery in the same hospital from 1999 to 2009. Results All the patients were firstly diagnosed as classic GCTB except for one case which was malignant pelvic GCTB. All patients received denosumab preoperatively and/or postoperatively, and the average number of medications was 8.43. According to the surgical patterns, patients were divided into intralesional surgery group (13 cases) and wide resection group (10 cases). The follow-up was 5-47 months(mean:27.30 months),recurrence was observed in 2 cases in the intralesional surgery group, none in the wide resection group. After drug administration, 13 cases were partial response, 7 cases were stable disease, the objective response rate was 65.0 % (13/20), and the histologically clearance rate of giant cells was 85.0 % (17/20). No case of osteonecrosis of the jaw was observed in this study, and all laboratory indicators were normal. The average postoperative MSTS-93 score was 26.87. Compared with pelvic GCTB patients who underwent surgical treatment from 1999 to 2009, in the intralesional surgery group, there was no significant difference in the recurrence rate [15.4 % (2/13) vs. 30.8 % (4/13), P = 0.514], but the limb function was significantly increased (P= 0.002). Conclusions Denosumab combined with surgery plays an important role in the multidisciplinary treatment of pelvic GCTB. The neoadjuvant strategy can reduce patient's intraoperative blood loss by shrinking the tumor size which makes the intralesional curettage surgery possible, and also diminishing the recurrence rate. But more attention should be paid to secondary malignant GCTB during the use of denousmab.

Expression features of osteoprotegerin, receptor activator of nuclear factor kappa-B and receptor activator of nuclear factor kappa-B ligand in avascular necrosis femoral head / 中国生化药物杂志

Objective To determine the protective effect of osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B (RANK) and RANK ligand (RANKL) on avascular necrosis femoral head.Methods Necrotic tissue or corresponding normal tissues were collected from 29 avascular necrosis femoral head patients.Quantitative Real TimePCR ( qPCR ) is used to evaluate mRNA expression of OPG , RANK and RANKL.OPG and RANKL protein levels were estimated by Western blot.Results The results of qPCR showed that the expression of OPG in the necrotic tissue was significantly higher than that in the normal tissue (4.56 ±0.37) (3.39 ±0.52) (P<0.05).The expression levels of RANKL mRNA in necrotic tissues and normal tissues were (0.86 ±0.11) and (0.31 ±0.08), respectively,the difference was statistically significant (P<0.05).The expression levels of RANK mRNA in necrotic tissues and normal tissmes were(0.87 ±0.12), (0.56 ±0.13) respectively,the difference was statistically significant (P<0.05).The ratio of RANKL/OPG and RANK/OPG in normal tissues were 0.69 and 0.52, respectively.In the necrotic tissues, RANKL/OPG and RANK/OPG ratios were 1.35 and 0.61, respectively.Results of Western blot showed that the expression of OPG in necrotic tissues was consistent with that in normal tissues.The expression of RANKL protein was detected in all samples , and the expression of RANKL protein in necrotic tissue and normal tissue was almost the same.RANK protein expression was not detected in all samples.Conclusion OPG, RANK and RANKL play important roles in progress of bone remodeling in necrotic area and in disturbance of bone homeostasis and might have an effect on bone destruction and subsequent collapse of hip joint.

Effect of Wenhua Juanbi Recipe () on expression of receptor activator of nuclear factor kappa B ligand, osteoprotegerin, and tumor necrosis factor receptor superfamily member 14 in rats with collagen-induced arthritis / 中国结合医学杂志

<p><b>OBJECTIVES</b>To study the effect of Wenhua Juanbi Recipe (, WJR) on expression of receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), and tumor necrosis factor receptor superfamily member 14 (TNFRSF14, also known as LIGHT) in rats with collagen-induced arthritis (CIA).</p><p><b>METHODS</b>CIA rats were generated by subcutaneous injection of bovine collagen type-II at the tail base. Sixty CIA rats were randomly assigned (10 animals/group) to: model, methotrexate (MTX)-treated (0.78 mg/kg body weight), and WJR-treated (22.9 g/kg) groups. Healthy normal rats (n=10) were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrifificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry.</p><p><b>RESULTS</b>Compared with the normal group, toe swelling degree was signifificantly increased in the model group (P<0.01). After treatment, toe swelling degree decreased signifificantly in the WJR and MTX groups compared with the model group (P<0.01). Compared with the normal group, expression of RANKL and LIGHT were signifificantly increased and OPG signifificantly decreased in peripheral blood and synovium of the model group (P<0.01). Conversely, RANKL and LIGHT expression were signifificantly reduced and OPG increased in the WJR and MTX groups compared with the model group (P<0.01). No statistically significant difference existed between WJR and MTX groups.</p><p><b>CONCLUSION</b>WJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.</p>

Effect of sumac extract on serum oxidative status, RANKL/OPG system and alveolar bone loss in experimental periodontitis in rats

Objectives Sumac (Rhus coriaria L.) is widely used spice which has several properties such as antioxidant, anti-inflammatory and antimicrobial. The purpose of this animal study was to evaluate the effects of sumac extract on levels of receptor activator of nuclear factor-kappa B ligand (RANKL), osteoprotegerin (OPG) expression, serum oxidative status, and alveolar bone loss in experimental periodontitis. Material and Methods Twenty-four Wistar rats were separated into three groups: non-ligated (NL, n=8), ligature only (LO, n=8), and ligature and treated with sumac extract (S, n=8) (20 mg/kg per day for 11 days). A 4/0 silk suture was placed around the mandibular right first molars subgingivally; after 11 days, the rats were sacrificed, and alveolar bone loss was histometrically measured. The detection of RANKL and OPG were immunohistochemically performed. Levels of serum total antioxidant status (TAS)/total oxidant status (TOS), and oxidative stress index (OSI) were also analyzed. Results Alveolar bone loss was significantly greater in the LO group compared to the S and NL groups (p<0.05). The number of inflammatory cell infiltrate (ICI) and osteoclasts in the LO group was significantly higher than that of the NL and S groups (p<0.05). The number of osteoblasts in the LO and S groups was significantly higher than that of the NL group (p<0.05). There were significantly more RANKL-positive cells in the LO group than in the S and NL groups (p<0.05). OPG-positive cells were higher in S group than in LO and NL groups (p<0.05). TOS and OSI levels were significantly reduced in S group compared to LO group (P<0.05) and TAS levels were similar in S and NL group (p>0.05). Conclusions The present study showed that systemic administration of sumac extract may reduce alveolar bone loss by affecting RANKL/OPG balance, TOS and OSI levels in periodontal disease in rats. .

Association between serum RANKL/OPG ratio and osteoporotic fracture in patients with rheumatoid arthritis / 中华全科医师杂志

Objective To investigate the value of serum receptor activator of nuclear factor kappa B ligand (RANKL)/osteoprotegrin (OPG) ratio in osteoporotic fracture (OPF) of patients with rheumatoid arthritis (RA).Methods Three hundred and eighty four RA patients with mean age of (49 ± 14) y (16-82) admitted in the First Affiliated Hospital of Anhui Medical University from 2010 to 2013 and 158 sex-and age-matched healthy subjects were enrolled in the study.OPF was diagnosed by X-ray examination and BMDs of femur and lumbar spine 2-4 (L2-4) were measured by dual energy X-ray absorptiometry.Levels of RANKL and OPG in the peripheral blood of 220 RA patients and 100 normal subjects were detected by ELISA method.Results Eighty-two cases of OPF was diagnosed in 384 RA patients (21.35％),the rate was higher than that in controls (3.80％,6/158,x2 =25.371,P ＜0.01).The peripheral blood levels of RANKL (0.150 ± 0.143 vs.0.101 ± 0.066,t =4.178,P ＜ 0.01),OPG (0.457 ± 0.293 vs.0.359 ±0.216,t=3.347,P=0.001) and ratio of RANKL/OPG (0.41 ±0.35 vs.0.34±0.20,t =2.111,P=0.036) in RA patients were significantly higher than those in control group.In comparison with normal controls,BMDs of all detected regions in RA were decreased significantly (P ＜0.01).The incidence of osteoporosis in RA (121/327,37％) was higher than that in normal controls (22/158,13.92％) (x2 =27.291,P ＜ 0.01).RA patients with OPF had higher age (t =4.377,P ＜ 0.01),longer duration of disease (t =2.612,P =0.009),higher RANKL level (t =3.554,P =0.001),higher RANKL/OPG ratio (t =2.651,P =0.010),higher health assessment questionnaires (HAQ) score (t =2.418,P =0.016),lower serum calcium level (t =2.183,P =0.030),lower hemoglobin level (t =2.125,P =0.036),higher Sharp score in hands X-ray examination (t =2.747,P =0.007),worse X-ray stage (x2 =7.856,P =0.049),higher glucocorticoid utilization rate (x2 =9.066,P =0.003) and higher incidence of osteoporosis (x2 =38.186,P ＜ 0.01),compared with patients without OPF.RA patients taking corticosteroids had higher incidence of osteoporosis (x2 =7.489,P =0.006) and higher incidence of OPF (x2 =9.066,P =0.003).Logistic regression analysis showed that age (OR =1.029,P =0.039,95％ CI:1.001-1.057)and the occurrence of osteoporosis (OR =3.159,P =0.001,95％ CI:1.562-6.385),RANKL/OPG ratio (OR =3.516,P =0.013,95 ％ CI:1.305-9.647) were risk factors for RA patients with OPF.Conclusion A higher incidence of OPF is prevalent in RA patients,and age,osteoporosis,taking glucocorticoids and RANKL/OPG ratio are risk factors for OPF in RA patients.

Effect of recombinant human osteoprotegerin on RANKL,OPG protein expression in alveolar bone tissue of rat with periodontitis / 重庆医学

Objective To investigate the effects of recombinant human osteoprotegerin(rhOPG) on RANKL ,OPG protein expression in alveolar bone tissue of rats with periodontitis to provide the experimental evidence for the application of rhOPG in pe‐riodontitis treatment .Methods Totally 22 Wistar rats were enrolled .The random number table was adopted to select two healthy rats as the healthy group .The rest 20 rats were selected as the experimental group for establishing the rat models of periodontitis , and then subdivided into the experimental control group (n=10) and rhOPG group (n=10) .Rats in the rhOPG group were locally injected by rhOPG 10 mg/kg at periodontal pocket gap of maxillary second molar ,while those in the experimental control group were injected by sterile water for injection at the same site and some volume .The streptavidin‐perosidase(SP) method was em‐ployed to detect the expression of RANKL ,OPG protein in alveolar bone tissue .Results Compared with the healthy group ,the ex‐pression levels of OPG in alveolar bone tissue of rats in the experimental group were lower with statistically significant difference (P0 .05) . Compared with the experimental control group ,the expression level of OPG protein in alveolar bone tissue of rats in the rhOPG group was significantly up‐regulated ,while that of RANKL protein was significantly down‐regulated(P<0 .05) .The OPG expres‐sion level after treatment in the rhOPG group was markedly enhanced ,while the RANKL expression level was reduced compared with before treatment ,the difference was statistically significant (P<0 .05) .Conclusion rhOPG may regulates the expression of RANKL and OPG in alveolar bone tissue of rats with periodontitis .