OBJECTIVES: The core pathology of osteoporosis lies in bone resorption exceeding bone formation; thus, promoting osteogenesis is a key therapeutic strategy. The osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) forms the biological basis of bone formation. Astragaloside IV (A-IV), a major active component of Astragalus membranaceus, is known to enhance osteogenesis, but its precise molecular mechanisms remain unclear. This study aims to investigate the effects of A-IV on the proliferation and osteogenic differentiation of BMSCs from osteoporotic rats and to elucidate its molecular mechanism through the regulation of microRNA-21 (miR-21) and Notch2 expression. METHODS: After 1 week of adaptive feeding, mature female SD rats were randomly divided into a sham-operated (Sham) group (n=4) and an ovariectomized (OVX) group (n=8) to establish an osteoporosis model. Twelve weeks after surgery, BMSCs were isolated from femoral bone marrow and cultured. Cells were divided into a S-BMSCs group (from Sham), an O-BMSCs group (from OVX), and an A-BMSCs group (from OVX-derived BMSCs treated with A-IV). S-BMSCs and O-BMSCs were induced for osteogenic differentiation using osteogenic induction medium, whereas A-BMSCs were treated with A-IV before induction. Flow cytometry was used to identify mesenchymal stem cell surface markers (CD29) and hematopoietic stem cell marker (CD34) to confirm BMSC characteristics. Cell proliferation was assessed using the methyl thiazolyl tetrazolium (MTT) assay. Alizarin red staining was performed to quantify calcium nodule formation, and alkaline phosphatase (ALP) activity assays were used to evaluate osteogenic differentiation. Real-time reverse transcription PCR (real-time RT-PCR) was used to detect changes in osteogenic-related genes, runt-related transcription factor 2 (Runx2) and osteopontin (OPN), as well as miR-21 expression. Western blotting was performed to assess Runx2, OPN, and Notch2 protein expression. RESULTS: Flow cytometry confirmed that O-BMSCs retained the phenotypic characteristics of mesenchymal stem cells. A-IV significantly enhanced the proliferation of BMSCs from osteoporotic rats (P<0.05), increased ALP activity, and upregulated the mRNA and protein expression of Runx2 and OPN (P<0.05). Bioinformatic and experimental analyses demonstrated that miR-21 directly targeted Notch2. A-IV treatment increased miR-21 expression while suppressing Notch2 protein expression and inhibiting activation of the Notch signaling pathway (P<0.05). CONCLUSIONS Astragaloside IV promotes the osteogenic differentiation of BMSCs derived from osteoporotic rats by upregulating miR-21 expression and inhibiting the key Notch signaling protein Notch2, thereby relieving the Notch2-mediated suppression of osteogenesis.
Animals ; Triterpenes/pharmacology* ; Saponins/pharmacology* ; Osteogenesis/drug effects* ; MicroRNAs/metabolism* ; Rats, Sprague-Dawley ; Female ; Cell Differentiation/drug effects* ; Mesenchymal Stem Cells/drug effects* ; Signal Transduction/drug effects* ; Osteoporosis/pathology* ; Rats ; Cells, Cultured ; Receptor, Notch2/metabolism* ; Receptors, Notch/metabolism* ; Ovariectomy ; Cell Proliferation/drug effects*

BACKGROUNDOsteoporosis is a common complication of chronic obstructive pulmonary disease (COPD). Recent clinical and biological researches have increasingly delineated the biomolecular pathways of bone metabolism regulation in COPD. We extended this work by examining the specific association and potential contribution of the osteoprotegerin (OPG)/receptor activator of nuclear factor-κB ligand (RANKL) axis to the pathogenesis of osteoporosis in advanced COPD. The aim of this study was to assess the relationships of serum OPG, RANKL, and tumor necrosis factor-alpha (TNF-μ) with bone turnover in men with very severe COPD.

METHODSPulmonary function, T-score at the lumbar spine (LS) and femoral neck (FN), serum OPG, RANKL, soluble receptor of tumor necrosis factor-alpha-I and II (sTNFR-I, sTNFR-II), osteocalcin (OC), and β-CrossLaps (βCL) levels were measured in 45 men with very severe stage COPD and 36 male non-COPD volunteers. COPD patients and healthy controls were compared using an independent t-test and Mann-Whitney U-test. The Pearson coefficient was used to assess the relationships between variables.

RESULTSOPG and OC were lower in male COPD patients than in control subjects whereas RANKL, serum βCL, TNF-μ, and its receptors were higher. OPG directly correlated with forced expiratory volume in 1 s (FEV1) % predicted (r = 0.46, P < 0.005), OC (r = 0.34, P < 0.05), LS (r = 0.56, P < 0.001), and FN T-score (r = 0.47, P < 0.01). In contrast, serum RANKL inversely associated with LS and FN T-score (r = -0.62, P < 0.001 and r = -0.48, P < 0.001) but directly correlated with βCL (r = 0.48, P < 0.001). In addition, OPG was inversely correlated with RANKL (r = -0.39, P < 0.01), TNF-μ (r = -0.56, P < 0.001), and sTNFR-I (r = -0.40, P < 0.01).

CONCLUSIONOur results suggest that serum OPG and RANKL levels are inversely associated with bone loss in men with advanced stage COPD.

Bone Density ; physiology ; Cross-Sectional Studies ; Humans ; Male ; Middle Aged ; Osteoporosis ; metabolism ; pathology ; Osteoprotegerin ; metabolism ; Pulmonary Disease, Chronic Obstructive ; metabolism ; pathology ; RANK Ligand ; metabolism ; Respiratory Function Tests ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the clinicopathologic characteristics of parathyroid carcinoma (PTC).

METHODSEleven cases of PTC encountered during the period from 1994 to 2012 were enrolled into the study. Forty cases of parathyroid adenoma (PA) were also retrieved for comparison. The clinical manifestations, laboratory results and pathologic features were analyzed, with literature review.

RESULTSThe main clinical manifestations of PTC included neck mass (11/11), hypercalcemia (11/11) and hyperparathyroidism (11/11). Most patients also had osteoporosis (10/11). In contrast, PA often manifested as hypercalcemia (40/40) and hyperparathyroidism (40/40). Histologic examination of PTC showed that the tumor cells contained clear to eosinophilic cytoplasm and separated by dense bands of fibrosis. The tumor mass was surrounded by thick fibrous capsule. Foci of capsular invasion and vascular permeation were identified at the tumor periphery in all cases. Cellular atypia was not conspicuous but mitotic figures and coagulative necrosis were easily identified. On the other hand, PA were composed of tumor cells with clear to eosinophilic cytoplasm, forming glands, trabeculae or nests. Most of them (35/40) had intact fibrous capsule. Mitotic figures were rarely encountered and tumor necrosis was absent. Immunohistochemical study showed that the tumor cells in PTC were positive for CK19 (11/11), chromogranin A (9/11), synaptophysin (7/11) and parathyroid hormone (11/11). They were negative for thyroglobulin, TTF-1 and calcitonin. The Ki-67 index was less than 10% (range = 2% to 9%). In contrast, the tumor cells in PA were positive (40/40) for CK19, chromogranin A, synaptophysin and parathyroid hormone. They were negative for thyroglobulin, TTF-1 and calcitonin. The Ki-67 index was less than 3%. Follow up-data were available in 9 cases of PTC (duration of follow up = 11 months to 224 months) and 7 of the patients were still alive. Follow up of all PA cases showed no evidence of recurrence.

CONCLUSIONSPTC is a rare malignant endocrine tumor presenting as neck mass. Histologic features suggestive of malignant behavior include presence of coagulative tumor necrosis and capsular/vascular invasion. It needs to be distinguished from other entities such as parathyroid adenoma, papillary thyroid carcinoma and medullary thyroid carcinoma.

Adenoma ; metabolism ; pathology ; Adult ; Carcinoma ; metabolism ; pathology ; Carcinoma, Neuroendocrine ; Carcinoma, Papillary ; Chromogranin A ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Hypercalcemia ; etiology ; Hyperparathyroidism ; etiology ; Immunohistochemistry ; Keratin-19 ; metabolism ; Male ; Middle Aged ; Osteoporosis ; etiology ; Parathyroid Hormone ; metabolism ; Parathyroid Neoplasms ; complications ; metabolism ; pathology ; surgery ; Synaptophysin ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology

Reports describing significant health risks due to inadequate vitamin D status continue to generate considerable interest amongst the medical and lay communities alike. Recent research on the various molecular activities of the vitamin D system, including the nuclear vitamin D receptor and other receptors for 1,25-dihydroxyvitamin D and vitamin D metabolism, provides evidence that the vitamin D system carries out biological activities across a wide range of tissues similar to other nuclear receptor hormones. This knowledge provides physiological plausibility of the various health benefits claimed to be provided by vitamin D and supports the proposals for conducting clinical trials. The vitamin D system plays critical roles in the maintenance of plasma calcium and phosphate and bone mineral homeostasis. Recent evidence confirms that plasma calcium homeostasis is the critical factor modulating vitamin D activity. Vitamin D activities in the skeleton include stimulation or inhibition of bone resorption and inhibition or stimulation of bone formation. The three major bone cell types, which are osteoblasts, osteocytes and osteoclasts, can all respond to vitamin D via the classical nuclear vitamin D receptor and metabolize 25-hydroxyvitamin D to 1,25-dihydroxyvitamin D to activate the vitamin D receptor and modulate gene expression. Dietary calcium intake interacts with vitamin D metabolism at both the renal and bone tissue levels to direct either a catabolic action on the bone through the endocrine system when calcium intake is inadequate or an anabolic action through a bone autocrine or paracrine system when calcium intake is sufficient.
Calcium/metabolism ; Fractures, Bone/metabolism/pathology ; Humans ; Osteoporosis/metabolism/pathology ; Protein Binding ; Receptors, Calcitriol/genetics/metabolism ; Vitamin D/analogs & derivatives/*metabolism

OBJECTIVETo evaluate the roles or effects of oviductus ranae (OR) or oviductus ranae eggs (ORE) in preventing and treating postmenopausal osteoporosis.

METHODSIn vivo experiment: Sixty female adult Wistar rats were randomly divided into 5 groups of 12. To provide an osteoporosis model 4 groups of rats were ovariectomized (OVX), with the 5th being sham operated. Medication commenced 7 days after the operation and lasted continuously for 12 weeks. Sham operated and OVX groups were given equivalent volumes of 5% Tween-80. The other three groups intragastrically received conjugated estrogens (CE), OR or ORE of the corresponding doses. At the 12th week, serum estrogen, bone gla protein (BGP), serum calcium, phosphorus, and alkaline phosphatase (ALP) were assayed; bone mineral densities (BMD) were measured and bone scanning was conducted; uteri were weighed, and weight, volume and length of the femoral bones were determined; and cortical thickness of femoral heads and area of bone trabecula were measured by image analyzer. In vitro experiment: Eighty 10-month old SD rats, with equal numbers of males and females, were randomly divided into 8 groups. Osteoblasts were isolated from neonatal rat calvariae, and the cells were exposed to various concentrations of serum from OR and ORE groups to study the impact of these sera on osteoblastic proliferation, ALP activity and mineralization. Osteoclastic numbers were determined using tartrate resistant acid phosphatase (TRAP).

RESULTSIn vivo experiment: The body weight of the four OVX groups increased significantly (P<0.01). Uterine weight of the CE group was the highest (P<0.01); Compared with the model group, estrogen level, BMD, bone scanning/bone imaging index weight of the femoral bones, cortical thickness of femoral heads in the OR and ORE groups increased significantly (P<0.05, P<0.01); femoral volume in the ORE group increased significantly (P<0.05); and the content of osteocalcin, phosphorus, and ALP in serum decreased significantly (P<0.05, P<0.01). In vitro experiment: Sera from OR and ORE groups had notable effects on the proliferation of osteoblasts (P<0.05 and P<0.01, repsectively) and stimulated the formation of calcium nodes (P<0.05, P<0.01), while the enhancement of ALP activity in osteoblasts was significant (P<0.05, P<0.01). The number of TRAP-positive cells was significantly reduced as well (P<0.01).

CONCLUSIONSOR and its eggs could effectively suppress OVX-induced osteoporosis in rats, and increase bone turnover possibly by both an increase in osteoblastic activity and a decrease in osteoclastic activity. The present study provides evidence that OR and its eggs could be considered a complementary and alternative medicine for the treatment of postmenopausal osteoporosis.

Acid Phosphatase ; metabolism ; Alkaline Phosphatase ; metabolism ; Animals ; Biomarkers ; blood ; Body Weight ; drug effects ; Bone Density ; drug effects ; Bone and Bones ; metabolism ; Calcification, Physiologic ; drug effects ; Cell Count ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Female ; Femur ; drug effects ; metabolism ; pathology ; Isoenzymes ; metabolism ; Male ; Materia Medica ; pharmacology ; therapeutic use ; Organ Size ; drug effects ; Osteoblasts ; drug effects ; enzymology ; pathology ; Osteoclasts ; drug effects ; enzymology ; pathology ; Osteoporosis ; blood ; drug therapy ; metabolism ; physiopathology ; Ovariectomy ; Ovum ; metabolism ; Rats ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase ; Uterus ; drug effects ; pathology

OBJECTIVETo investigate the preventive effects and possible underlying mechanism of different extracts of Kanggushu () on osteoporosis in ovariectomized rats.

METHODSOne hundred and sixtyfive female SD rats were divided into 11 groups: control, sham, model, Xianling Gubao Capsule (), nilestriol, Kanggushu aqueous extract high-, medium-, and low-dose and suet extract high-, medium-, and low-dose groups. The osteoporosis model was made by ovariectomizing the rats. The latter 8 groups were administered intragastricly with Xianling Gubao Capsule, nilestriol, Kanggushu aqueous extract and suet extract for 12 weeks, respectively, while the other 3 groups were administered orally saline. The whole body bone mineral density, bone mineral content, organ coefficient of uterus, serum estradiol and alkaline phosphatase contents, blood calcium, phosphorus, interleukin 6 and bone Gla-protein levels after treatment were monitored. Additionally, three-point bending test of femur, HE staining, and scanning electron microscope were performed to explore the pharmacodynamics and underlying mechanisms.

RESULTSIn comparison with ovariectomized rats of model group, Kanggushu aqueous extract high-dose resulted in an increased bone mineral density, bone mineral content and organ coefficient of uterus, improved estradiol level, and improved maximum load and structural stiffness (P<0.05 or P<0.01). Two-dimensional and three-dimensional trabecular structure was also observed under HE staining and scanning electron microscopy, and the number and thickness of trabecular bone in Kanggushu aqueous extract high-dose group was significantly increased compared to the model group, while the lipid droplets in bone marrow cavity were significantly less. However, there were no significant differences in blood calcium, total serum alkaline phosphatase and bone Gla protein among different treatment groups. Overall, the osteoprotective effects of Kanggushu aqueous extract were comparable to those of nilestriol and were significantly more effective than those of Xianling Gubao Capsule.

CONCLUSIONThe preventive effects of Kanggushu aqueous extract might be partly due to the increased estradiol level, accelerated restoration of bone trabecular reticulate structure, and accordingly increased bone mineral density in osteoporosis rats.

Alkaline Phosphatase ; metabolism ; Animals ; Biomechanical Phenomena ; drug effects ; Body Weight ; drug effects ; Bone Density ; drug effects ; Calcium ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Estradiol ; blood ; Female ; Femur ; drug effects ; pathology ; physiopathology ; ultrastructure ; Interleukin-6 ; metabolism ; Organ Size ; drug effects ; Osteocalcin ; metabolism ; Osteoporosis ; blood ; drug therapy ; pathology ; physiopathology ; Ovariectomy ; Phosphorus ; metabolism ; Rats ; Rats, Sprague-Dawley ; Uterus ; drug effects ; pathology

OBJECTIVETo evaluate the role of sclerostin in bone loss of postmenopausal Chinese women with type 2 diabetes mellitus.

METHODSThe postmenopausal patients suffering from type 2 diabetes mellitus and age, body mass index, and duration of menopause matched healthy controls were enrolled into this cross-sectional study according to criteria of inclusion and exclusion. The serum sclerostin level and bone mineral density of the anterior-posterior lumbar spine (L1-L4), femoral neck, and total hip were determined by using a quantitative sandwich ELISA kit and dual X-ray absorptiometry, respectively. Meanwhile, the clinical and laboratory indexes of bone mineral metabolism were analyzed. Associations between serum sclerostin level and bone mineral density as well as bone turnover markers were evaluated by linear regression analysis.

RESULTSFinally, 265 postmenopausal women with type 2 diabetes and 225 non-diabetic women were recruited in the diabetic group and control group, respectively. Serum sclerostin level of the diabetic group was significantly higher than that of the control group (48.2±19.4 vs. 37.2±18.6 pmol/L, P<0.001) and was increased with age in both groups (diabetic group, r=0.374, P<0.001; control group, r=0.312, P<0.001). In type 2 diabetes patients, serum sclerostin concentration was positively correlated with hemoglobin A1c level (r=0.237; P=0.021). Biochemical bone turnover markers, intact parathyroid hormone and bone-specific alkaline phosphatase, were negatively associated with serum sclerostin level (r=-0.138, P=0.078 and r=-0.265, P<0.001). Conversely, the positive correlation between sclerostin and C-terminal cross-linking telopeptide of type I collagen was found in diabetic patients (r=0.354, P<0.001). Serum sclerostin levels of the diabetic group were positively correlated with bone mineral density of the lumbar spine, femoral neck, and total hip (r=0.324, 0.367, and 0.416, respectively; all P<0.001).

CONCLUSIONSSclerostin might participate in the pathogenesis of bone loss of type 2 diabetes. The high sclerostin level might serve as a marker of increased osteocyte activity in postmenopausal patients with type 2 diabetes mellitus.

Aged ; Alkaline Phosphatase ; blood ; Asian Continental Ancestry Group ; Biomarkers ; blood ; Bone Morphogenetic Proteins ; blood ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; blood ; epidemiology ; Female ; Genetic Markers ; Hemoglobin A ; metabolism ; Humans ; Middle Aged ; Osteocytes ; metabolism ; pathology ; Osteoporosis, Postmenopausal ; blood ; epidemiology ; Parathyroid Hormone ; blood ; Retrospective Studies

The effect of the anti-inflammatory flavonoid chrysin on osteogenesis was determined in preosteoblast MC3T3-E1 cells. Results demonstrated that chrysin could induce osteogenic differentiation in the absence of other osteogenic agents. Chrysin treatment promoted the expression of transcription factors (Runx2 and Osx) and bone formation marker genes (Col1A1, OCN, and OPN) as well as enhanced the formation of mineralized nodules. During osteogenic differentiation, chrysin preferentially activated ERK1/2, but not JNK nor the p38 MAPKs. Further experiments with inhibitors revealed the co-treatment of U0126, PD98059, or ICI182780 (a general ER antagonist) with chrysin effectively abrogated the chrysin-induced osteogenesis and ERK1/2 activation. Thus, the effect of chrysin on osteogenesis is ERK1/2-dependent and involves ER. Therefore, chrysin has the significant potential to enhance osteogenesis for osteoporosis prevention and treatment.
Animals ; Cell Differentiation ; drug effects ; Cell Line ; Enzyme Activation ; drug effects ; Flavonoids ; pharmacology ; therapeutic use ; Mice ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; Osteoblasts ; drug effects ; pathology ; Osteogenesis ; drug effects ; Osteoporosis ; drug therapy ; pathology ; Phosphorylation ; drug effects ; Receptors, Estrogen ; metabolism

OBJECTIVETo observe the bone mineral density changes of coal workers' pneumoconiosis.

METHODSWe chose 150 cases of One-Triple coal workers pneumoconiosis in Jincheng Coal Mining Group, all of workers were male, of 55-years old-80 years old, an average of 67 years old. 10 years of age to grouping, Whole body bone mineral density and T value were measured by body dual-energy X-ray absorptiometry. We analyzed the BMD changes bone loss, osteoporosis occurrence.

RESULTSThe BMD of six parts were not declined obviously in One stage of coal workers' pneumoconiosis; the BMD of Chest bone, pelvis and spine were declined obviously in two stage of coal workers' pneumoconiosis; the BMD of six parts were declined obviously in Triple stage of coal workers' pneumoconiosis; The occurrence rate of bone loss was significantly higher in Two and Triple coal workers pneumoconiosis. The occurrence rate of osteoporosis was significantly higher in Triple coal workers pneumoconiosis.

CONCLUSIONWith the increase in the severity of coal workers' pneumoconiosis, the BMD of six parts were declined, The occurrence rate of bone loss osteoporosis was significantly higher.

Aged ; Aged, 80 and over ; Anthracosis ; epidemiology ; metabolism ; pathology ; Bone Density ; Case-Control Studies ; Coal Mining ; Humans ; Male ; Middle Aged ; Osteoporosis ; epidemiology ; pathology

OBJECTIVETo investigate the relationship between the level of sex hormones and the loss of bone mass in aging male rats.

METHODSThirty male Sprague-Dawley rats were equally divided into five age groups and sacrificed at 35, 70, 160, 700 and 800 postnatal days (PD) , followed by measurement of the % Tb x Ar, Tb x Th, Tb x N and Tb x SP by bone histomorphometry and detection of the levels of serum testosterone (T) and estradiol (E2) by radioimmunoassay. The relationship between the changes of the T and E2 levels and those of bone histomorphometry was analyzed.

RESULTST and E2 levels were closely correlated with the bone mass in the aging male rats. The changes in T and E2 levels were simultaneous with those in the bone mass with the growth of the rats. T, E2, % Tb x Ar and Tb x N reached the peak in the 70 and 160 PD groups, and markedly decreased in the 700 PD group except Tb x Th and Tb x SP. The T levels in the 35, 70, 160, 700 and 800 PD groups were (118.53 +/- 18.35) ng/dl, (345.49 +/- 54.63) ng/dl, (368.83 +/- 60.03) ng/dl, (61.15 +/- 21.12) ng/dl and (60.35 +/- 19. 27) ng/dl, changing simultaneously with the E2 levels, which were (10.35 +/- 1.82) pg/ml, (16.92 +/- 3.13) pg/ml, (17.20 +/- 2.51) pg/ml, (5.87 +/- 2.34) pg/ml and (5.53 +/- 2.48) pg/ml, respectively. The metrological parameters of the bone structure in the five groups were as follows, Tb x Ar: (19.52 +/- 2.23)%, (26.28 +/- 2.18) %, (28.37 +/- 1.21) %, (15.62 +/- 1.68) % and (14.21 +/- 0.89) %; Tb x Th: (35.45 +/- 1.63) microm, (50.13 +/- 3.58) microm, (60.23 +/- 8.25) microm, (75.62 +/- 9.72) microm and (78.78 +/- 11.21) microm; Tb x N: (5.98 +/- 1.21) n/mm, (8.07 +/- 0.86) n/mm, (8.30 +/- 1.22) n/mm, (2.63 +/- 1.35) n/mm and (2.48 +/- 1.62) n/mm; Tb x SP: (126.34 +/- 18.15) microm, (136.26 +/- 15.27) microm, (261.08 +/- 76.43) microm, (323.12 +/- 78.12) microm and (330.23 +/- 50.20) microm.

CONCLUSIONChanges in the levels of sex hormones are closely correlated with those of bone mass. Both testosterone and estradiol are essential for bone development and bone mass maintenance.

Aging ; Animals ; Bone Density ; Bone Development ; Bone and Bones ; metabolism ; Cytoskeleton ; metabolism ; pathology ; Estradiol ; blood ; Male ; Osteoporosis ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood