Traditional Chinese Medicine in Prevention and Treatment of Osteonecrosis of Femoral Head Based on OPG/RANK/RANKL Signaling Pathway： A Review / 中国实验方剂学杂志

Osteonecrosis of the femoral head （ONFH） is a painful and debilitating disease caused by impaired blood supply to the femoral head and cellular and tissue degeneration， leading to gradual destruction of the bone structure and progressive collapse of the femoral head. The main pathological mechanism of ONFH is the disruption of the balance between bone absorption and the reconstruction of new bone， resulting from microcirculation damage and decreased cellular tissue ability. This imbalance leads to biomechanical changes and accelerates the pathological progression of ONFH. In the early stages， clinical manifestations may not be obvious， mainly presenting as pain or discomfort in the hip or groin area， which can be relieved after rest. In the later stage of the disease， pain intensifies， and limb shortening， lower limb weakness， difficulty walking， or limping may occur. Currently， western medicine commonly uses osteogenic agents， anticoagulants， and artificial joint replacement for treatment， but there are also many issues such as prosthesis loosening and infection. Research has shown that traditional Chinese medicine （TCM） treatment of ONFH takes a holistic approach and employs multi-functional， multi-target， and multi-system Chinese medicine therapies， ensuring the safety and effectiveness of the treatment. The osteoprotegerin （OPG）/receptor activator of nuclear factor-κB （RANK）/RANK ligand （RANKL） signaling pathway plays a crucial role in maintaining the dynamic balance of bone remodeling. TCM treatments utilize this pathway to promote apoptosis of osteoclasts， reduce bone resorption， and accelerate bone formation， thereby playing an important role in the prevention and treatment of ONFH. This paper reviewed the role of OPG/RANK/RANKL signaling pathway and related cytokine expression in ONFH by reviewing relevant literature in China and abroad and research status of Chinese medicinal monomers， Chinese medicinal formulations， and combinations with physical therapy in increasing osteoblast secretion， promoting OPG expression， enhancing cytokine expression levels， and inhibiting osteoclast activity for the prevention and treatment of ONFH. This paper is expected to provide new ideas and directions for TCM in the prevention and treatment of ONFH.

Traditional Chinese Medicine Intervenes in Signaling Pathways Associated with CKD-MBD： A Review / 中国实验方剂学杂志

Traditional Chinese medicine （TCM） has certain advantages in the treatment of chronic kidney disease-mineral and bone disorder （CKD-MBD）. In recent years， there have been many studies on the treatment of CKD-MBD by Chinese medicinal compounds and monomers. As revealed by literature retrieval， the research on the mechanism of Chinese medicine in intervening in signaling pathways related to CKD-MBD was mainly based on self-made Chinese medicinal compounds， and the action pathways involved fibroblast growth factor 23/Klotho （FGF23/Klotho） signaling pathway， Wnt/β-catenin signaling pathway， receptor activator of nuclear factor-κB/receptor activator of nuclear factor-κB ligand/osteoprotegerin （RANK/RANKL/OPG） system， and other signaling pathways. TCM can improve calcium and phosphorus metabolism and bone metabolism disorder， and regulate inflammatory reaction， oxidative stress， apoptosis， and autophagy by regulating this series of signaling pathways for the treatment of CKD-MBD. This paper introduced the research results of these signaling pathways and the mechanism of TCM in the treatment of CKD-MBD in order to provide ideas and references for the related research of Chinese medicine in the treatment of CKD-MBD.

Effects of bone marrow mesenchymal stem cell transplantation on bone metabolism and RANKL/OPG/TRAF6 signaling pathway in osteoporotic rats / 中华内分泌外科杂志

Objective:To investigate the effect of bone marrow derived mesenchymal stem cells (BMSC) transplantation on bone metabolism and its mechanism in ovariectomized osteoporosis rats.Methods:Forty clean SD female rats aged 7 weeks were divided into 4 groups according to the random number table method: sham operation group, model group, the transplantation group, positive control group, in addition to control the rest of the group were performed bilateral oophorectomy build osteoporosis rats model, after 2 months of model establishment, rats in transplantation group were injected with 80 μl/kg PBS solution containing bone marrow mesenchymal stem cells through tail vein, rats in sham operation group and model group were injected with the same amount of PBS solution through tail vein, and rats in positive control group were given Xianlinggubao (0.5 g/100 g) by gavage every day. Serum and femur were collected 14 days after treatment. Hematoxylin and eosin staining (HE) was used to observe the histopathological changes of femur. Micro-CT was used to measure bone mineral density and bone parameters. The expression levels of osteocalcin, osteoprotegerin, alkaline phosphatase and insulin-like growth factor 1 were detected by enzyme-linked immunosorbent assay (ELISA) kit. The serum levels of calcium, phosphorus and magnesium were measured by spectrophotometer. The protein expressions of RANKL, OPG, TRAF6 and NF-KB1 in femur of each group were detected by Western blot.Results:Compared with the sham operation group, the bone mineral density (BMD) of the model group was decreased by (0.28±0.01) g/cm 3, bone volume fraction (BMD) was decreased by (0.28±0.01) g/cm 3. BV/TV) decreased by (19.73±2.02) %, trabecular thickness (Tb.Th) decreased by (0.082±0.008) mm, trabecular number (Tb.N) decreased by (1.60±0.17) mm -1 and trabecular separation/spacing (Tb.Sp) increased (0.273±0.024) mm, osteoprotegerin (489.49±55.29) ng/L, alkaline phosphatase (229.13±15.05) U/L, insulin-like growth factor-1 (236.64±14.32) μg/L, and osteocalcin were decreased (1.866±0.109) μg/L, calcium (11.98±1.09) mg/dl, phosphorus (6.85±0.68) mg/dl, and magnesium decreased (0.62±0.04) mg/dl) , the relative expression level of RANKL increased (1.05±0.09) , the relative expression level of OPG decreased (0.58±0.08) , the relative expression level of RANKL increased (0.74±0.10) , and the relative expression level of NF-kB1 increased (1.01±0.11) ( P<0.05) ; bone mineral density, bone mineral density, bone mineral density BMD (0.38±0.04 g/cm 3, BV/TV (26.73±2.74) %, Tb.Th (0.094±0.006) mm, Tb.N (2.67±0.09) mm-1 and Tb.Sp were decreased (0.241±0.026) mm) , osteoprotegerin (720.09±67.41) ng/L, alkaline phosphatase (269.48±14.15) U/L, insulin-like growth factor 1 (IGF-1) decreased (335.95±24.13) μg/L, and osteocalcin increased (1.392±0.153) μg/L, calcium (7.12±0.53) mg/dl, phosphorus (4.54±0.32) mg/dl, magnesium (0.87±0.08) mg/dl. RANKL relative expression level increased (0.59±0.05) , OPG relative expression level decreased (0.97±0.10) , RANKL relative expression level increased (0.45±0.06) , NF-kB1 relative expression level increased (0.72±0.06) ( P<0.05) ;bone mineral density, bone mineral density, bone mineral density BMD (0.36±0.05) g/cm 3, BV/TV (28.72±3.20) %, Tb.Th (0.096±0.011) mm, Tb.N (2.85±0.24) mm -1 Tb.Sp was basically unchanged (0.241±0.027) mm, osteoprotegerin was decreased (716.78±36.90) ng/L, alkaline phosphatase was basically unchanged (270.65±18.59) U/L, and insulin-like growth factor 1 was decreased (336.94±17.50) μg/L, osteocalcin (1.377±0.101) μg/L, calcium (7.13±0.80) mg/dl, phosphorus (4.58±0.71) mg/dl, and magnesium (0.89±0.04) remained unchanged mg/dl, the relative expression level of RANKL increased (0.55±0.08) , the relative expression level of OPG decreased (0.98±0.13) , the relative expression level of RANKL was basically unchanged (0.40±0.05) , and the relative expression level of NF-kB1 increased (0.65±0.09) ( P<0.05) . Conclusion:Bone marrow mesenchymal stem cell transplantation can improve osteoporosis in ovariectomized rats by regulating bone metabolism and serum levels of calcium, phosphorus and magnesium, which may be related to RANKL/OPG/TRAF6 pathway.

Age Estimation Using Panoramic Radiography and Lateral Cephalogram—A Comparative Study

Context: Age estimation allows the identification of living individuals, dead victims and estimation of age in children using maxillofacial radiographs. Aim: To compare age estimation by mandibular third molar developmental stages using modified Demirjian’s method on OPG with age estimation by mandibular linear dimensions on lateral cephalogram. Setting and Design: A total of 200 randomly selected subjects (100 males and 100 females), with age range from 9 to 20 years, and 200 Digital Orthopantomograms and 200 Digital Lateral Cephalograms were utilized for the study. Methods and Material: Radiographs were taken with Kodak 8000C Digital Panoramic and Cephalometric machine operating at 60–90 kvp, with exposure time of 8–18 s at 2–15 mA, with an inbuilt magnification factor. A flat screen Compaq TFT?LCD monitor was used to view the OPG images. Linear mandibular dimensional measurements were made on each Digital Lateral Cephalogram using Trophy Dicom Software. Statistical Analysis Used: Regression analysis and regression coefficient for establishing gender?specific equation were observed. Evaluation of results and statistical analysis was carried by using Student’s ‘t’ test. For all test, ‘P’ value of 0.05 or less was utilized for measuring the degree of significance. Intra?observer variability was detected using reliability analysis. Results: The accuracy of age estimation by OPG was 93.8% and by lateral cephalogram was 79.7%. Conclusion: The OPG analysis is more reliable than the cephalometric parameters.

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.

Experimental study on the regulation of subchondral bone plate osteoprotegerin/receptor activator of nuclear factor kappa-B ligand system by berberine to retard the development of osteoarthritis in rabbits / 中国骨伤

OBJECTIVE@#To investigate the effect of intra-articular berberine injection on the structural remodeling of subchondral bone plate and osteoprotegerin/receptor activator of nuclear factor kappa-B ligand(OPG/RANKL) system expression in rabbits with osteoarthritis(OA).@*METHODS@#Forty 12-month-old male rabbits with an average of(2.73±0.18) kg of body weight, underwent left anterior cruciate ligament transection(ACLT), and were divided into berberine group and placebo groups after operation, 20 rabbits in each group. The berberine group received intra-articular injection of 100 μmol/L berberine 0.3 ml every week for 6 weeks. In placebo group, the same dose of 0.9% sodium chloride injection was injected into the left knee joint cavity every week for 6 weeks. Another 20 12-month-old male rabbits, weighing (2.68±0.18) kg, underwent sham operation on the left knee joint without intra-articular injection intervention (sham operation group). On the last day of the sixth week after operation, three groups of animals were sacrificed to obtain knee joint specimens. The femoral medial condyle samples were obtained for histological evaluation of cartilage and subchondral bone, Mankin scoring system was used to evaluate articular cartilage structure. Image-Pro Plus(IPP) software was used to evaluate subchondral bone plate bone volume(BV), bone volume/total volume(BV/TV), trabecular circumference(TC), mean trabecular thickness (Tb.Th). Real-time quantitative reverse transcription polymerization Enzyme chain reaction(reverse transcription-polymerase chain reaction, RT-PCR) was used to detect the mRNA expression levels of OPG and RANKL in subchondral bone tissue at 6 weeks after operation.@*RESULTS@#The cartilage structure evaluation showed that the surface of cartilage tissue in the sham operation group was smooth and flat, and the safranin coloration was full in the full thickness of the cartilage;the cartilage tissue in the berberine group showed uneven surface layer, and the staining of safranin O was mildly decreased;the surface layer fibrosis was seen in placebo group, Safranin O faded significantly. The Mankin score in the berberine group was lower than that in placebo group(P<0.01), but higher than that in sham operation group(P<0.01). The structural evaluation of subchondral bone plate showed that the trabecular bone in sham-operated group was densely arranged;after berberine intervention, the trabeculae were closely arranged;the subchondral bone trabeculae in placebo group were relatively sparse, and the distance between trabeculae was wider. Subchondral bone plate IPP software evaluation showed that BV, BV/TV, TC, Tb.Th in berberine group were higher than those in placebo group(P<0.01), BV, BV/TV, TC, Tb.Th in berberine group were higher than those in placebo group(P<0.01), while lower than the sham operation group (P<0.01). PCR test results showed that the expression of OPG mRNA in the berberine group was significantly higher than that in placebo group(P<0.01), and OPG mRNA in the berberine group was lower than that in sham operation group (P<0.01). There was no significant difference in mRNA expression of RANKL among three groups(P>0.05);the ratio of OPG/RANKL in berberine group was higher than that in placebo group(P<0.01), but lower than that in sham operation group(P<0.01).@*CONCLUSION@#Intra-articular injection of berberine can effectively inhibit the resorption of subchondral bone in the early stage of OA and delay the development of the disease. The specific mechanism may be that berberine maintains the balance of OPG/RANKL system by up-regulating the expression of OPG gene in subchondral bone.

Effects of intrathecal injection of resveratrol on sense of pain and OPG/RANK/RANK-L signal pathway in rats with bone cancer pain / 中国药房

OBJECTIVE To study the effects of intrathecal injection of resveratrol on sense of pain and OPG/RANK/RANK-L signal pathway in rats with bone cancer pain. METHODS After treatment via intrathecal tube ，the rats were randomly divided into sham operation group ，model group ，resveratrol low-dose ，medium-dose and high-dose groups （0.25，1，2 mg/kg），with 8 rats in each group. Except for sham operation group ，other groups were given Walker 256 breast cancer cell suspension on the right hind limb tibia to induce bone cancer pain model. After 12 days of modeling ，intrathecal administration was performed in resveratrol groups. The model group and sham operation group received the same volume of dimethyl sulfoxide ，10 μL，once a day ，for 5 days. The paw withdrawal mechanical threshold （PWMT）and paw withdrawal thermal latency （PWTL）were measured at 3 d，5 d after administration. The serum levels of tumor necrosis factor α（TNF-α），interleukin 6（IL-6），IL-1β and chemokine ligand 2 （CCL2）were determined after last medication ，and the protein expressions of RANK ，RANK-L and OPG in tibia were also determined. RESULTS Compared with sham operation group ，PWMT and PWTL were significantly decreased in the model group 12 days after operation （P＜0.05）. Compared with model group ，PWMT and PWTL were significantly increased in resveratrol medium-dose and high-dose groups at 3 d and 5 d after administration （P＜0.05）. Compared with model group ，the levels of TNF-α，IL-6，IL-1β and CCL2 in serum were significantly decreased in resveratrol middle-dose and high-dose groups at 5 d after administration（P＜0.05）；the expression of IL- 6 in serum was significantly decreased in resveratrol low-dose group （P＜0.05）； the protein expressions of RANK-L and RANK in tibia were significantly decreased ，while the protein expression of OPG was increased significantly in resveratrol middle-dose group （P＜0.05）. CONCLUSIONS Resveratrol by intrathecal injection can relieve the inflammation caused by bone cancer pain of rats ， the mechanism of which may be associated with inhibiting the activity of OPG/RANK/RANK-L signal pathway.

Caracterização de polimorfismos nos genes OPG, RANK e RANKL em uma população brasileira / Characterization of OPG, RANK and RANKL gene polymorphisms in a Brazilian population

Introdução: o sistema RANKL (receptor-ativador do fator nuclear-ligante κB)/RANK (receptor ativador do NF-kB)/OPG (osteoprotegrina) Introdução: o sistema OPG (osteoprotegrina)/RANK (receptor ativador do NF-kB)/RANKL (receptor-ativador do fator nuclear-ligante κB) regula os processos fisiológicos e patológicos da remodelação óssea. Polimorfismos genéticos nos genes OPG, RANK e RANKL têm sido associados a doenças, em diferentes populações. Objetivo: Descrever a frequência e o potencial regulatório dos polimorfismos do sistema OPG, RANK e RANKL em uma população brasileira; avaliar o seu potencial como marcadores genéticos informativos de ancestralidade; comparar com patologias associadas em outras populações. Metodologia: neste estudo, 506 indivíduos adultos, participantes de uma coorte acometidos de asma e periodontite, tiveram o DNA genômico extraído e genotipado, utilizando-se a plataforma Illumina. As plataformas NCBI, RegulomeDB, Haploview 4.2 e rSNPBase foram consultadas e utilizadas para análises. Resultados e Discussão: os polimorfismos mais frequentes na população estudada foram o rs3102724 no gene OPG, com frequência de menor alelo (MAF) de 46%; o rs4941129 em RANK, MAF 50%; e o rs9525641 em RANKL, MAF 46%. Os rs3134063 (1f) em OPG, rs17069898 (1f) em RANK e rs2200287 (1d) em RANKL apresentaram maior impacto funcional. Em OPG e RANK, nove polimorfismos se caracterizaram como marcadores genéticos informativos de ancestralidade, com predomínio nas populações YRI (africanos) e CEU (europeus). Os nove polimorfismos, com função intrônica, apresentaram MAF entre 2 a 46% na população-alvo e foram associados a patologias do metabolismo ósseo em outras populações. Conclusão: polimorfismos dos genes estudados se mostraram frequentes na população estudada e tiveram seus alelos mais frequentes associados a doenças em populações ancestrais. Sugere-se que sejam realizados mais estudos.

Introduction: The OPG (osteoprotegerin)/ RANK (NF-kB activating receptor)/ RANKL (nuclear-binding factor κB receptor-activating system regulates the physiological and pathological processes of bone remodeling. Genetic polymorphisms (SNPs) in OPG, RANK and RANKL genes have been associated with diseases in different populations. Objective: Describe the regulatory frequency and potential of SNPs in OPG, RANK and RANKL in a Brazilian population; assess their potential as informative genetic markers of ancestry; compare with pathologies associated with these polymorphisms in other populations. Methods: in this study, 506 adult individuals, participating in a cohort involving asthma and periodontitis, had genomic DNA extracted and genotyped using the Illumina platform. The NCBI, RegulomeDB, Haploview 4.2 and rSNPBase platforms were consulted and used for analysis. Results and discussion: the most frequent polymorphisms in the studied population were the rs3102724 in the OPG gene, with the lowest allele frequency (MAF) of 46%; rs4941129 in RANK, MAF 50% and rs9525641 in RANKL, MAF 46%. The rs3134063 (1f) in OPG, rs17069898 (1f) in RANK and rs2200287 (1d) in RANKL, had greater functional impact. In OPG and RANK, 9 SNPs were characterized as informative genetic markers of ancestry, predominantly in YRI (African) and CEU (European) populations. These 9 SNPs, with intronic function, presented MAF between 2 and 46% in our population, and were associated with pathologies in bone metabolism in other populations. Conclusion: SNPs of the studied genes were found to be frequent in the studied population and had their most frequent alleles associated with diseases in ancestral populations. It is suggested that further studies be carried out

Effect of Kidney-tonifying Method on OPG/RANK/RANKL Signaling Pathway in Postmenopausal Osteoporosis Animal Model： A Meta-analysis / 中国实验方剂学杂志

Objective:As the problem of global aging intensifies，postmenopausal osteoporosis （PMOP） has become a global health problem among females. At present，the commonly used biological agents have been proved not suitable for long-term use due to multiple adverse reactions. Several Meta-analyses have confirmed the good safety and effectiveness of kidney-tonifying method against PMOP，but its therapeutic mechanism remains unclear. The purpose of this Meta-analysis was to evaluate the effect of kidney-tonifying method on osteoclastogenesis inhibitory factor（OPG）/receptor activator of nuclear transcription factor （NF）-<italic>κ</italic>B （RANK）/receptor activator of NF-<italic>κ</italic>B ligand （RANKL） signaling pathway in PMOP animal model，so as to provide an experimental basis for the treatment of PMOP with kidney-tonifying method. Method:The related articles were retrieved from PubMed，Ovid Medline，Embase，China National Knowledge Infrastructure （CNKI），Chongqing Weipu Database for Chinese Technical Periodicals （VIP），and Wanfang Data Knowledge Service Platform with the retrieval time set from their inception to January 2020. The quality of each included article was evaluated using the SYRCLE's risk of bias tool. Then RevMan 5.3 was utilized for Meta-analysis according to the Cochrane systematic review methodology. Result:Thirty-two studies involving 619 rats were included. The quality score of these studies ranged from 3 to 5 points. The results of the Meta-analysis indicated obvious advantages of kidney-tonifying method in increasing bone mineral density （BMD）［standardized mean difference （SMD）=2.01，95% confidence interval（CI）=1.50-2.52，<italic>P</italic><0.000 01］），serum OPG level （SMD=3.33，95% CI=2.59-4.07，<italic>P</italic><0.000 01），and OPG mRNA expression （SMD=11.81，95% CI=7.49-16.13，<italic>P</italic><0.000 01），promoting OPG protein production （SMD=4.95，95% CI=3.09-6.81，<italic>P</italic><0.000 01），reducing serum RANKL（SMD=-4.88，95% CI=-6.01--3.75，<italic>P</italic><0.000 01） and RANK levels （SMD=-7.30，95% CI=-9.53--5.07，<italic>P</italic><0.000 01），and down-regulating RANKL （SMD=-6.22，95%CI=-8.95--3.49，<italic>P</italic><0.000 01） and RANK mRNA （SMD=-3.18，95% CI=-6.19--0.18，<italic>P</italic><0.05） expression and RANKL protein expression in bone tissue （SMD=-3.99，95% CI=-5.47--2.50，<italic>P</italic><0.000 01）. Conclusion:The kidney-tonifying method has been proved to possess potential advantages in regulating the balance of OPG/RANK/RANKL signaling pathway in PMOP animal model. Nevertheless，more large-sample sized，properly designed，and high-quality animal experiments are still needed for further verification.

Therapeutic Effect of Guizhi Shaoyao Zhimutang on Osteoporosis in Rats Based on RANKL/OPG / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect of Guizhi Shaoyao Zhimutang （GSZT） on the osteoporosis （OP） in SD female rats induced by ovariectomy （OVX） combined with glucocorticoid injection and its related mechanisms. Method:The rats were divided into a sham operation group and an experimental group for OVX. One week later， the experimental rats were divided into an OP model group， a calciferol （positive control， 0.1 mg·kg^-1） group， and low（0.8 g·kg^-1）， medium（1.6 g·kg^-1）， and high-dose （3.2 g·kg^-1） GSZT groups. Except for those in the sham operation group， the rats received an intramuscular injection of dexamethasone （1 mg·kg^-1） twice per week for six weeks. After the rats were treated correspondingly for eight weeks， the rats were sacrificed for thighbone sample collection. The computer microtomography （Micro CT） was used to analyze the parameters of bone mineral density （BMD）， bone volume （BV）， tissue volume （TV）， structure model index （SMI）， connectivity density （Conn.D）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， trabecular pattern factor （Tb.Pf）， and trabecular separation （Tb.Sp）. In addition， hematoxylin-eosin （HE） and Masson staining was used for the histopathological examination of the thighbone. Tartrate-resistant acid phosphatase （TRAP） staining was used to analyze osteoclasts in bone tissues. The enzyme-linked immunosorbent assay （ELISA） was used to determine matrix metalloproteinase-9 （MMP-9）， cathepsin K （CTSK）， and TRAP5b in bone tissues， and Western blot was carried out to determine receptor activator of NF-kappaB ligand （RANKL）， osteoprotegerin （OPG）， alkaline phosphatase （ALP） in bone tissues. Result:Compared with normal group， the number of bone trabeculae in cancellous part of OP model rats decreased significantly， the spacing widened， the thickness became thinner， and the continuity was poor. Compared with model group， GZST medium and high-dose groups could improve the changes of femoral bone trabecular cancellous bone. Masson staining showed that compared with the normal group， the growth plate thickness of the distal femur epiphysis was thinner and the area of new bone was smaller in the OP model group， while the reduction of new bone was inhibited in the high dose group of calcitol and GZST. TRAP staining results showed that the number of osteoclasts in OP group was significantly increased compared with the normal group， and the number of osteoclasts in medium-high dose GZST group was reduced. Micro CT results showed that compared with normal group， BMD， BV， Tb.N， Tb.Th and Conn.D of OP rats were decreased， SMI and Tb.Sp were significantly increased（<italic>P</italic><0.01）. Compared with model group， medium and high dose of GZST could improve the degree of osteoporosis in OP rats， BV， Tb.N were significantly increased， SMI， Tb.Sp were significantly decreased （<italic>P</italic><0.01）. ELISA results revealed that compared with the sham operation group， the OP model group showed decreased ALP content （<italic>P</italic><0.01） and increased MMP-9， TRAP5b， and CTSK （<italic>P</italic><0.01）. Compared with the OP model group， the high-dose GSZT group showed increased ALP content （<italic>P</italic><0.01） and decreased MMP-9， TRAP5b， and CTSK （<italic>P</italic><0.01）. Western blot results demonstrated that compared with the OP model group， the calciferol group and the high-dose GSZT group showed inhibited expression of the RANKL protein （<italic>P</italic><0.05）， and increased OPG expression （<italic>P</italic><0.05）. Conclusion:The findings suggest that GZST has a potential therapeutic effect on OP caused by OVX combined with glucocorticoids in rats， and the underlying mechanism is associated with the regulation of RANKL/OPG in bone tissues. This study is expected to lay a preliminary foundation for its clinical application and modern preparation development.

A panoramic study of the morphology of mandibular condyle in a sample of population from Basrah City / Estudio panorámico de la morfología del cóndilo mandibular en una muestra de población de la ciudad de Basora

SUMMARY: The study aims to evaluate the morphological variation of mandibular condyle This analysis was composed of 450 digital orthopantomograms taken for routine investigation to assess the morphology of 900 condylar heads. Condylar morphology was classified into four types (oval, bird beak, diamond and crooked finger). Most common occurring shape were evaluated, symmetry of condylar shape and most common combinations of the condylar shapes present in population were assessed. Out of 900 mandibular condyle, type 1(oval) of condyle was the most commonly appearing shape (56 %), followed by type 3 (diamond), type 2(bird beak) and finally type 4 (crooked finger). Symmetry was seen in 74.2 % of the total sample with the oval-oval was the most commonly occurring. Routine use of orthopantomography can assist dentist in assessment of condylar morphology alterations. Oval shape was the most common in all age groups in males and females. More studies on Iraqi population evaluating mandibular condyle are needed to provide standardized values for mandibular anatomical parts.

RESUMEN: El objetivo del presente estudio fue evaluar la variación morfológica del cóndilo mandibular. El análisis se realizó en 450 ortopantomogramas digitales en una investigación de rutina para evaluar la morfología de 900 cabezas condilares. La morfología condilar se clasificó en cuatro tipos (ovalada, pico de pájaro, diamante y dedo torcido). Se evaluó la forma más común de ocurrencia, la simetría de la forma condilar y las combinaciones más comunes en la población. De 900 cóndilos mandibulares, el tipo 1 (ovalado) de cóndilo apareció con mayor frecuencia (56 %), seguido del tipo 3 (diamante), el tipo 2 (pico de pájaro) y finalmente el tipo 4 (dedo torcido). Se observó simetría en el 74,2 % de la muestra total, siendo el óvalo-óvalo el que se presentó con mayor frecuencia. El uso rutinario de la ortopantomografía puede ayudar al dentista a evaluar las alteraciones de la morfología condilar. La forma ovalada fue la más común en todos los grupos etarios en hombres y mujeres. Se necesitan más estudios sobre la población iraquí que evalúen el cóndilo mandibular para proporcionar valores estandarizados para evaluar la anatomía mandibular.

Ocorrência de espécies do gênero Eimeria Schneider, 1875, em cordeiros confinados / Occurrence of Eimeria Schneider, 1875 species in feedlot lambs

Este trabalho descreve as espécies do gênero Eimeria Schneider, 1875, que ocorreram em um confinamento de cordeiros, bem como as dinâmicas da eliminação de oocistos no ambiente, a correlação com o ganho de peso médio diário (GMD) e as variáveis climáticas, durante nove semanas. Cento e quatro cordeiros de diversas raças e cruzas, com aproximadamente 60 dias de vida, foram confinados e submetidos a pesagens e avaliações clínicas e coprológicas periódicas. Amostras de fezes com mais de 500 oocistos de Eimeria por grama de fezes (OoPG) foram separadas para esporulação e identificação das espécies. Entre os oocistos avaliados, foram identificadas as espécies: E. parva, E. crandallis, E. ovinoidalis, E. weybridgensis, E. bakuensis, E. marsica, E. ahsata, E. granulosa, E. pallida e E. faurei. Eimeria crandallis foi a mais frequente, presente em 44 das 58 amostras avaliadas, enquanto E. parva foi a mais abundante nas contagens individuais. Nenhum dos animais apresentou quadro de eimeriose, e coeficientes negativos foram encontrados nas correlações OoPG vs. GMD (-0,075) e OoPG vs. pluviosidade (-0,1164), enquanto para OoPG vs. temperatura foi encontrado coeficiente positivo (0,2914). Animais positivos para a eliminação de oocistos apresentaram infecção mista nas avaliações semanais, com até sete espécies parasitando um mesmo cordeiro.(AU)

This study describes the Eimeria Schneider, 1875 species that occurred in a lamb feedlot, as well as the dynamics of oocyst output in the environment and its correlation with daily weight gain (DWG) and climatic variables during nine weeks. One hundred and four lambs of various breeds and crossbreeds, at approximately 60 days old, were placed in a feedlot and submitted to periodic weighing and clinical and coprological evaluations. Fecal samples presenting more than 500 Eimeria spp. oocysts per gram (OPG) were separated for sporulation, and oocysts were identified at species level. Among evaluated oocysts, the following species were identified: E. parva, E. crandallis, E. ovinoidalis, E. weybridgensis, E. bakuensis, E. marsica, E. ahsata, E. granulosa, E. pallida and E. faurei. Eimeria crandallis was the most frequent one, being identified in 44 of the 58 evaluated samples, while E. parva was the more abundant in individual counts in most weeks. None of the animals presented clinical signs of eimeriosis and negative correlation coefficients were found for OPG vs DWG (-0.075) and OPG vs rainfall, as for OPG vs temperature this coefficient was positive. Animals shedding oocysts presented mixed infection, with up to seven species parasitizing the same lamb.(AU)

Impact of supplementation with different protein sources on the parasitological profile of ovine matrices and development of lambs / Impacto da suplementação com diferentes fontes protéicas no perfil parasitológico de matrizes ovinas e no desenvolvimento de cordeiros

The aim was to evaluate the impact of supplementation with different protein sources from agro-industrial production on the parasitological profile of ovine matrices and the development of their offspring. The protein sources used were: soybean bran, cotton pie, babassu pie, and coconut pie. A total of 60 ovine matrices were used, with confirmed pregnancy and supplemented in the final third of gestation at 0.5% of live weight in pastures of deferred Marandu grass. The evaluations occurred from the final third of gestation and in the lactation phase until weaning. The data collected in the matrices were: counting of eggs per gram of feces (EPG) and identification of infective larvae (Faecal test), evaluation of the degree of anemia by the FAMACHA© method, body weight (kg), and body condition score (BCS). In the lambs, the biometric measurements were taken from the 30 days of life until weaning. In Faecal test, the prevalence was 98% of Haemonchus spp. There was no effect of the protein sources for EPG (3476.0) and FAMACHA©. However, in the mats supplemented with coconut pie were observed the lowest body weights (36.9 kg) and lower BCS (2.7). Babassu, coconut and cotton cakes, and soybean meal used as a protein source in the supplementation of ovine matrices kept in deferred pastures of Marandu grass at the 0.5% level of live weight maintain the production indexes consistent with the physiology stage of the matrices even with a high endo-parasitic load.

O objetivo foi avaliar o impacto da suplementação com diferentes fontes proteicas da produção agroindustrial no perfil parasitológico das matrizes ovinas e no desenvolvimento de seus filhotes. As fontes de proteína utilizadas foram: farelo de soja, torta de algodão, torta de babaçu e torta de coco. Foram utilizadas 60 matrizes ovinas, com prenhez confirmada e suplementadas no terço final da gestação com 0,5% do peso vivo em pastagens de capim Marandu diferido. As avaliações ocorreram desde o terço final da gestação e na fase de lactação até o desmame. Os dados coletados nas matrizes foram: contagem de ovos por grama de fezes (OPG) e identificação de larvas infectantes (teste fecal), avaliação do grau de anemia pelo método FAMACHA ©, peso corporal (kg) e escore de condição corporal (ECC). Nos cordeiros, as medidas biométricas foram realizadas a partir dos 30 dias de vida até o desmame. No teste fecal, a prevalência foi de 98% de Haemonchus spp. Não houve efeito das fontes de proteína para OPG (3476,0) e FAMACHA ©. No entanto, nas matrizes suplementadas com torta de coco foram observados os menores pesos corporais (36,9 kg) e menorECC (2,7). Torta de Babaçu, coco, algodão e farelo de soja utilizados como fonte protéica na suplementação de matrizes ovinas mantidas em pastos diferidos de capim Marandu no nível de 0,5% de peso vivo mantêm os índices de produção consistentes com o estágio fisiológico das matrizes, mesmo com uma carga endo-parasitária alta.

The effect of cigarette smoking and low-level laser irradiation in rank/rankl/opg expression

Abstract The objective of this study was to investigate the effects of low-level laser therapy (LLLT) and cigarette smoke on alveolar socket osteoclastogenesis signaling after tooth extraction, in rats. Sixty male Wistar rats were randomly assigned to four groups with 15 animals each: Control Group (with right maxillary molar extraction - ME), Experimental I (with ME and LLLT), Experimental II (with ME and cigarette smoke) and Experimental III group (with ME, LLLT and cigarette smoke). Euthanasia was performed at 3, 7 and 14 days postoperative. qRT-PCR was used to evaluate expression of Tnfrsf11a (RANK), Tnfsf11 (Rankl) and Tnfrsf11b (OPG). Data were submitted to statistical analysis using two-way ANOVA followed by Bonferroni test (α=0.05). There was an upregulation of RANK, RANKL and OPG genes over all the time of healing in Exp I group compared to control group. Exp II group showed a decreased expression of all genes over time, whereas Exp III genes expression were higher than Exp II values but lower than Control and Exp I values over time. The results of this study concluded that the LLLT had a positive effect, whereas cigarette smoke had a negative effect on RANK, RANKL and OPG gene expression in bone remodeling process.

Resumo O objetivo deste estudo foi investigar os efeitos da terapia a laser de baixo nível (LLLT) e a fumaça de cigarro na sinalização da osteoclastogênese do alvéolo após extração dentária, em ratos. Sessenta ratos Wistar machos foram distribuídos aleatoriamente em quatro grupos com 15 animais cada: Grupo de controle (com extração do molar superior direito (EM), Experimental I (EM e LLLT), Experimental II (EM e fumaça de cigarro) e Grupo Experimental III (EM, LLLT e fumaça de cigarro). A eutanásia foi realizada aos 3, 7 e 14 dias após a extração. O qRT-PCR foi utilizado para avaliar a expressão de Tnfrsf11a (RANK), Tnfsf11 (RANKL) e Tnfrsf11b (OPG). Os dados foram submetidos à análise estatística usando two-way ANOVA seguido do teste de Bonferroni (α=0,05). Houve um aumento na expressão dos genes RANK, RANKL e OPG ao longo do tempo de cicatrização no grupo Exp I em comparação com o grupo controle. O grupo Exp II mostrou uma expressão diminuída de todos os genes ao longo do tempo, enquanto a expressão dos genes do grupo Exp III foi superior aos valores observados em Exp II, mas inferiores aos valores do grupo Controle e Exp I ao longo do tempo. Os resultados deste estudo concluíram que o LLLT tem um efeito positivo, enquanto a fumaça de cigarro possui efeito prejudicial na expressão gênica de RANK, RANKL e OPG no processo de remodelação óssea.

Effects of Curcumin on Bone Metabolism Balance in Ovariectomized Osteoporosis Model Rats Based on OPG/ RANKL Signaling Pathway / 中国药房

OBJECTIVE：To investigate the effects of curcumin on bone metabolism balance in ovariectomized osteoporosis model rats ，and to investigate its potential mechanism. METHODS ：Totally female Wistar rats were randomly divided into blank group（group A ），model group （group B ），estradiol group [group C （positive control ），estradiol valerate 50 μg（/ kg·d）]， curcumin low-dose ，medium-dose and high-dose groups [group D-F ，55，110，165 μg（/ kg·d）]，with 15 rats in each group. Except for group A ，other rats were ovariectomized to establish osteoporosis model. After modeling ，group A and B were given normal saline intrgastrically ，and administration groups were given relevant medicine intrgastrically 30 mL/kg，once a day ，for consecutive 12 weeks. The contents of serum bone metabolism markers [BALP ，CBF-α1，CTX-Ⅰ，PINP and OC] were determined by ELISA. The bone mineral density （BMD）and trabecular structure indexes [relative bone volume fraction （BV/TV），trabecular number （Tb.N），trabecular thickness （Tb.Th），connectivity density （Conn. D ），trabecular separation （Tb.Sp）and structure model index （SMI）] were determined by micro CT imaging system. The mRNA and protein expression of OPG and RANKL in hypothalamus and femur were determined by RT-PCR and Western blotting assay. RESULTS ：Compared with group A ，the contents of serum bone metabolism markers ，BMD，BV/TV，Tb.N，Tb.Th，Conn.D，mRNA and protein expression of OPG were decreased significantly in group B ，while Tb.Sp ，SMI，mRNA and protein expression of RANKL were increased significantly @qq.com （P＜0.05）. Compared with group B ，the contents of serum bone metabolism markers ，BMD，BV/TV，Tb.N，Tb.Th，Conn.D，mRNA and protein expression of OPG were increased significantly in administration groups ；Tb.Sp，SMI，mRNA and protein expression of RANKL were decreased significantly ，in a dose-dependent manner among curcumin groups （P＜0.05 or P＜0.01）. CONCLUSIONS ：Curcumin can improve the level of bone metabolism，increase BMD ，improve the trabecular microstructure and inhibit bone absorption in ovariectomized osteoporosis model rats. Its mechanism may be related to the regulation of OPG/RANKL signaling pathway.

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.

Effects of psoralen corylifolia extract on bone mineral density and bone biomechanics in osteoporosis rats / 中国组织工程研究

BACKGROUND: Long-term use of dexamethasone as a glucocorticoid will destroy the dynamic balance of osteogenesis and osteoclastation, reduce bone mineral density, damage bone biomechanics. A regulation of receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) protein pathway may be an approach to improve bone mineral density and bone biomechanics in dexamethasone-induced osteoporosis rats. OBJECTIVE: To investigate the effects of psoralen corylifolia extract on bone mineral density and bone biomechanics in dexamethasone-induced osteoporosis rats based on RANKL/OPG pathway. METHODS: Osteoporosis models were established in Wistar rats, SPF grade, using intramuscular injection of dexamethasone. Lentiviral vectors containing OPG gene interference fragments at concentration of 1×107 TU/mL were selected for subsequent experiments. The model rats were randomly divided into model group, no-load group (lentivirus empty vector), OPG silencing group (lentivirus vector containing OPG gene interference fragment), psoralen corylifolia extract group, psoralen corylifolia extract+OPG silencing group, with 12 rats in each group. Another 12 rats were selected as the control group. After drug treatment, bone mineral density of the left femur in rats was measured by bone densitometer; the elastic modulus, maximum load and yield load of the right femur of rats were measured by mechanical testing machine; the mineral salt content in the femur of rats was determined; the levels of RANKL and OPG in the serum of rats were detected by ELISA kit; and the expression levels of RANKL and OPG in bone tissues of rats were detected by western blot. The study protocol was approved by the Animal Ethics Committee of Qinghai University Medical School with an approval No. 2017081501. RESULTS AND CONCLUSION: (1) Bone mineral density, modulus of elasticity, maximum load, yield load, bone mineral salt content, serum OPG level and OPG protein expression in bone tissues were measured, which were lower in the model group than the control group, lower in the OPG silencing group than the model group, higher in the psoralea corylifolia extract group than the model group, and higher in the psoralen corylifolia extract+OPG silencing group than the OPG silencing group, but lower than the psoralea corylifolia extract group (all P 0.05). (4) All these findings indicate that psoralea corylifolia extract can increase bone mineral density and improve bone biomechanics in dexamethasone-induced osteoporosis rats, possibly by up-regulating the expression of OPG and down-regulating the expression of RANKL.

The role of OPG/RANKL/RANK signaling pathway in the pathogenesis of giant-cell tumor of bone / 中国组织工程研究

BACKGROUND: Studies have shown that osteoprotegerin/receptor activator of nuclear factor κB ligand/nuclear factor κB receptor activator (OPG/RANKL/RANK) signaling pathway has a certain correlation with the pathogenesis of giant-cell tumor. Controlling the OPG/RANKL/RANK signaling pathway to affect the interaction between osteoblasts and osteoclasts can play a certain therapeutic role in giant-cell tumor of bone. OBJECTIVE: Ton introduce the relationship between the OPG/RANKL/RANK signaling pathway and the pathogenesis of giant-cell tumor of bone, and to summarize and discuss the new advances of the OPG/RANKL/RANK signaling pathway in the pathogenesis of giant-cell tumor of bone. METHODS: PubMed, Web of Science, and WanFang databases were searched for relevant articles published from 2001 to 2019 using the keywords of “OPG/RANKL/RANK, giant cell tumor of bone, pathogenesis, signal pathway, bone metabolism" in English and Chinese, respectively. A total of 53 articles were finally included for analysis and discussion after removal of old and repeated literatures. RESULTS AND CONCLUSION: OPG inhibits the proliferation and differentiation of osteoclasts, reduces the activity of mature osteoclasts, and blocks the binding of RANKL to RANK. RANKL binds to RANK on the surface of osteoclast progenitor cells to promote the differentiation and proliferation of osteoclast progenitor cells, thus accelerating osteoclast progression. After binding to RANKL receptor, RANKL activates signal factors such as nuclear factor-κB to promote the proliferation, differentiation and activation of osteoclasts, and to regulate the transcription and expression of related genes. Therefore, the OPG/RANKL/RANK is associated with the pathogenesis of giant-cell tumor.

Evaluation of Serum RANKL and OPG Concentrations in Patients with Periodontitis in Saudi Arabia

Objective: To investigate RANKL and OPG serum concentrations and the RANKL/OPG ratio in patients diagnosed withchronic periodontitis and attending dental school clinics in Makkah, Saudi Arabia. Methods : Patients seeking dentaltreatment at the Faculty of Dentistry Clinic, Umm Al Qura University were divided into periodontitis and control groupsbased on their clinical findings. Periodontitis was diagnosed in patients with a PD >4mm or CAL >3mm at more than 30%of the examined sites, and the radiographic evidence of alveolar bone loss was > 30% of teeth. RANKL and OPG serumconcentrations (pg/ml) and RANKL/OPG ratio were determined. Results: Forty-three patients, including 32 females and 11males with a mean age of 30 +/_ 8.48, were studied. Thirty patients were diagnosed with periodontitis, and 13 were in thecontrol group. The patients with periodontitis had significantly higher RANKL (31.8 +/_ 27.5 vs. 18.7 +/_ 14.2 pg/ml,p=0.04) and OPG concentrations (58.0 +/_ 23.2 vs. 45.3 +/_ 10.0 pg/ml, p=0.03) than the control patients. The patients withperiodontitis that smoked had significantly lower serum RANKL concentrations than those that did not smoke (37.8 +/_ 10.0vs. 14.8 +/_ 10.0, p=0.0057 pg/ml). The RANKL/OPG serum ratio was higher in non-smokers than in smokers (0.6 vs. 0.3,p=0.017). The patients with generalized advanced periodontitis had significantly higher serum RANKL concentrations thanthose with the localized disease (56.8 +/_ 42.0 vs. 22.0 +/_ 12.8 pg/ml, p=0.03). Serum RANKL/OPG ratio was increased inpatients with generalized disease (0.96 vs. 0.39, p=0.036). Serum RANKL/OPG ratio was also correlated with the amount ofCAL (r = 0.459, p=0.04). Conclusions: Serum RANKL concentrations were increased in patients with periodontitis, in nonsmokers, and in patients with the generalized advanced disease.

Effects of drynaria total flavonoids on mandible structure, serum E2 and OPG/RANKL balance in ovariectomized rats / 西安交通大学学报(医学版)

Objective: To explore the effects of drynaria total flavonoids on serum estrogen (E2) level, OPG/RANKL balance and bone microarchitecture of mandible in a rat model of bone loss induced by ovariectomy. Methods: We randomly divided 40 three-month-old female Sprague-Dawley rats into 5 groups (n=8 per group): sham-operation group (Sham), ovariectomized group (OVX), low-dose drynaria total flavonoids group (GB-L), high-dose drynaria total flavonoids group (GB-H) and estradiol valerate group (EV). The mandibular osteoporosis model was established by removing the rats' ovaries. After 12 weeks the rats were killed, and the serum estrogen levels were measured. OPG and RANKL were detected using ELISA method. Therapeutic effect of drynaria total flavonoids was observed by pathomorphology and histomorphometry of the mandibular molar sections. Results: Compared with those in the sham group, the mandible of rats in OVX group was loosely changed, the level of E2 decreased significantly (P<0.001), and OPG/RANKL decreased significantly (P<0.001). Compared with OVX group, the mandible microstructure of rats in EV group got a good repair, while the serum E2 level and OPG/RANKL increased significantly (both P<0.001). Compared with OVX group, the percentage of trabecular bone (%Tb.Ar) and OPG/RANKL increased in all the GB groups (P<0.001, P=0.029), (P<0.001, P=0.007), while GB-H group also had increased thickness of trabecular bone (Tb.Th) and serum OPG levels (P=0.019, P<0.001), reduced trabecular spacing (Tb.Sp)(P=0.001), but no effect on serum E2 level (P=0.432, P=0.459). Conclusion: Drynaria total flavonoids can inhibit the destruction of the mandibular trabecular structure, thus slowing down the mandibular resorption process. However, they cannot improve E2 concentration. The mechanism is realized by adjusting the balance of OPG/RANKL.