Association of bone turnover biomarkers with severe intracranial and extracranial artery stenosis in type 2 diabetes mellitus patients

BACKGROUND Intracranial and extracranial artery stenosis is associated with cerebral infarction.Vascular calcification and atherosclerosis are the main causes of stenosis and major risk factors for cardiovascular and cerebrovascular events in patients with type 2 diabetes mellitus(T2DM).Bone turnover biomarkers(BTMs)are associated with vascular calcification,atherosclerosis,glucose,and lipid metabolism.AIM To investigate the association of circulating BTM levels with severe intracranial and extracranial artery stenosis in patients with T2DM.METHODS For this cross-sectional study including 257 T2DM patients,levels of the BTMs serum osteocalcin(OC),C-terminal cross-linked telopeptide of type I collagen(CTX),and procollagen type I N-peptide were measured by electrical chemiluminescent immunoassay,and artery stenosis was assessed by color Doppler and transcranial Doppler.Patients were grouped according to the existence and location(intracranial vs.extracranial)of artery stenosis.Correlations between BTM levels,previous stroke,stenosis location,and glucose and lipid metabolism were analyzed.RESULTS T2DM patients with severe artery stenosis had a higher frequency of previous stroke and levels of all three tested BTMs(all P<0.05)than patients without.Some differences in OC and CTX levels were observed according to the location of artery stenosis.Significant associations were also observed between BTM levels and some glucose and lipid homeostasis parameters.On multivariate logistic regression analysis,all BTMs were significant predictors of artery stenosis in T2DM patients with and without adjustment for confounding factors(all P<0.001),and receiver operating characteristic curve analysis demonstrated the ability of BTM levels to predict artery stenosis in T2DM patients.CONCLUSION BTM levels were found to be independent risk factors for severe intracranial and extracranial artery stenosis and were differentially associated with glucose and lipid metabolism in patients with T2DM.Therefore,BTMs may be promising biomarkers and potential therapeutic targets for artery stenosis.

Overexpression of Dlx2 enhances osteogenic differentiation of BMSCs and MC3T3-E1 cells via direct upregulation of Osteocalcin and Alp

Genetic studies have revealed a critical role of Distal-homeobox (Dlx) genes in bone formation,and our previous study showed that Dlx2 overexpressing in neural crest cells leads to profound abnormalities of the craniofacial tissues.The aim of this study was to investigate the role and the underlying molecular mechanisms of Dlx2 in osteogenic differentiation of mouse bone marrow stromal cells (BMSCs) and pre-osteoblast MC3T3-E1 cells.Initially,we observed upregulation of Dlx2 during the early osteogenesis in BMSCs and MC3T3-E1 cells.Moreover,Dlx2 overexpression enhanced alkaline phosphatase (ALP) activity and extracellular matrix mineralization in BMSCs and MC3T3-E1 cell line.In addition,micro-CT of implanted tissues in nude mice confirmed that Dlx2 overexpression in BMSCs promoted bone formation in vivo.Unexpectedly,Dlx2 overexpression had little impact on the expression level of the pivotal osteogenic transcription factors Runx2,Dlx5,Msx2,and Osterix,but led to upregulation of Alp and Osteocalcin (OCN),both of which play critical roles in promoting osteoblast maturation.Importantly,luciferase analysis showed that Dlx2 overexpression stimulated both OCN and Alp promoter activity.Through chromatin-immunoprecipitation assay and site-directed mutagenesis analysis,we provide molecular evidence that Dlx2 transactivates OCN and Alp expression by directly binding to the Dlx2-response cis-acting elements in the promoter of the two genes.Based on these findings,we demonstrate that Dlx2 overexpression enhances osteogenic differentiation in vitro and accelerates bone formation in vivo via direct upregulation of the OCN and Alp gene,suggesting that Dlx2 plays a crucial role in osteogenic differentiation and bone formation.

Modulation of Isoflavones on Bone-nodule Formation in Rat Calvaria Osteoblasts in vitro

Objective To observe the effects of two main isoflavones, daidzein and genistein on the bone-nodule formation in rat calvaria osteoblasts in vitro. Methods Osteoblasts obtained from newborn Sprague-dawley rat calvarias were cultured for several generations. The second generation cells were cultured in Minimum Essential Medium supplemented with ascorbic acid and Na-beta-glycerophosphate for several days, in the presence of daidzein and genistein, with or without the estrogen receptor antagonist ICI 182780. Number of nodules was counted at the end of the incubation period (day 20) by staining with Alizarin Red S calcium stain. The release of osteocalcin, as a marker of osteoblast activity, was also determined on day 7 and day 12 during the incubation period. Results Compared with the control, the numbers of nodules were both increased by incubation with daidzein and genistein. 17β-estradiol was used as a positive control and proved to be a more effective inducer of the increase in bone-nodules formation than daidzein and genistein. The release of osteocalcin into culture media was also increased in the presence of daidzein and genistein, as well as 17β-estradiol on day 7 and day 12 (day 12 were higher). The estrogen receptor antagonist ICI 182780 completely blocked the genistein- and 17β-estradiol-induced increase of nodule numbers and osteocalcin release in osteoblasts. However, the effects induced by daidzein could not be inhibited by ICI 182780. Conclusion These findings suggest that geinistein can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts. The effects, like those induced by 17β-estradiol, are mediated by the estrogen receptor dependent pathway. Daidzein also can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts, but it is not, at least not merely, mediated by the estrogen receptor dependent pathway.

Osteocalcin as a hormone regulating glucose metabolism

The number of patients with osteoporosis and diabetes is rapidly increasing all over the world. Bone is recently recognized as an endocrine organ. Accumulating evidence has shown that osteocalcin, which is specifically expressed in osteoblasts and secreted into the circulation, regulates glucose homeostasis by stimulating insulin expression in pancreas and adiponectin expression in adipocytes, resulting in improving glucose intolerance. On the other hand, insulin and adiponectin stimulate osteocalcin expression in osteoblasts, suggesting that positive feedforward loops exist among bone, pancreas, and adipose tissue. In addition, recent studies have shown that osteocalcin enhances insulin sensitivity and the differentiation in muscle, while secreted factors from muscle, myokines, regulate bone metabolism. These findings suggest that bone metabolism and glucose metabolism are associated with each other through the action of osteocalcin. In this review, I describe the role of osteocalcin in the interaction among bone, pancreas, brain, adipose tissue, and muscle.

Bone fragility in type 2 diabetes mellitus

The number of patients with osteoporosis or type 2 diabetes mellitus(T2DM)is increasing in aging and westernized societies.Both disorders predispose elderly people to disabling conditions by causing fractures and vascular complications,respectively.It is well documented that bone metabolism and glucose/fat metabolism are etiologically related to each other through osteocalcin action and Wnt signaling.Bone fragility in T2DM,which is not reflected by bone mineral density(BMD),depends on bone quality deterioration rather than bone mass reduction.Thus,surrogate markers are needed to replace the insensitivity of BMD in assessing fracture risks of T2DM patients.Pentosidine,the endogenous secretory receptor for advanced glycation endproducts,and insulin-like growth factor-I seem to be such candidates,although further studies are required to clarify whether or not these markers could predict the occurrence of new fractures of T2DM patients in a prospective fashion.

Serum levels of undercarboxylated osteocalcin are related to cardiovascular risk factors in patients with type 2 diabetes mellitus and healthy subjects

AIM To determine a potential relationship between serum undercarboxylated(uc OC) concentration and cardiovascular risk factors in type 2 diabetes(T2D) patients and healthy subjects(HS).METHODS A cross-sectional study was conducted on 140 subjects classified into two groups, 70 with T2D and 70 HS. Medical history and physical examination with anthropometric measurements were obtained from all subjects. Body fat percentage was determined by bioelectrical impendency analysis. Serum uc OC concentration was determined by enzyme immunoassay,while serum levels of insulin and hsC RP were obtained using high sensitivity enzyme-linked immunosorbent assay. Insulin resistance was determined using the homeostasis model assessment-IR. Lipid profile [triglycerides,total cholesterol(TC), high-density lipoproteins(HDL-c),low density lipoproteins(LDL-c), very low-density lipoproteins] was determined by spectrophotometry and standard formulas when applicable. RESULTS The T2D patient group showed significantly higher values of waist circumference, waist-to-hip ratio, systolic blood pressure(SBP), diastolic blood pressure(DBP),current smoking, and alcohol use when compared to the HS group(P < 0.05). We observed a significantly lower serum ucO C concentration in T2D than in HS(1.5 ± 1.4vs 2.3 ± 1.8, P < 0.05). In the whole study population,ucO C concentration was inversely correlated with body mass index(BMI)(r =-0.236, P < 0.05), fasting plasma glucose(r =-0.283, P < 0.01) and HDL-c(r =-0.255,P < 0.05); and positively correlated with LDL-c/HDL-c ratio(r = 0.306, P < 0.05) and TC/HDL-c ratio(r =0.284, P < 0.05). In the T2D group, serum uc OC concentration was inversely correlated with BMI(r =-0.310, P < 0.05) and body-fat percentage(r =-0.311,P < 0.05), and positively correlated with DBP(r = 0.450,P < 0.01). In HS group a positive correlation between serum levels of uc OC and SBP(r = 0.277, P < 0.05)was observed. CONCLUSION Serum uc OC is a potential marker for cardiovascular risk in Mexicans because it is related to adiposity parameters, blood pressure and lipid profile.

Effects of Anastrozole Combined with Shuganjiangu Decoction on Osteoblast-like Cell Proliferation, Differentiation and OPG/RANKL mRNA Expression

Objective： To investigate the effects of anastrozole combined with Shuganjiangu decoction on osteoblast-like cells. Methods： Human osteoblast-like cells MG-63 were cultured and divided into four groups control, anastrozole, Shuganjiangu decoction （SGJGD）, and anastrozole combined with SGJGD. Cell proliferation was investigated by M-IF assay. Alkaline phosphatase （ALP） and osteocalcin, the indicators of cell differentiation, were evaluated by p-nitrophenyl- phosphate method and radioimmunoassay, respectively. Gene expressions of ALP, osteocalcin, osteoprotegerin （OPG） and receptor activator of nuclear factor kappa B ligand （RANKL） were examined by real-time PCR. Results： As evidenced by MTT assay, cell proliferation of MG-63 was inhibited by anastrozole, but stimulated with treatment of SGJGD alone and combined with anastrozole （P〈O.01）. Compared with control group, ALP activity was increased by the treatment of SGJGD alone and combined with anastrozole （P〈0.01）. Also, osteocalcin secretion was enhanced with the treatment of SGJGD single and combination with anastrozole （P〈O.05）. In the real-time PCR assay, gene expressions of ALP and osteocalcin were significantly increased （P〈0.01 for ALP, P〈0.05 for osteocalcin） by the treatment of SGJGD and anastrozole combined with SGJGD, but the expression of RANKL was decreased （P〈O.05）. Moreover, anastrozole combined with SGJGD upregulated gene expression of OPG （P〈O.01）. Conclusion： SGJGD may alleviate the injury effects of anastrozole on MG-63 cells through adjusting bone formation and resorption indicators.

Bone Remodeling and Energy Metabolism:New Perspectives

Bone mineral, adipose tissue and energy metabolism are interconnected by a complex and multilevel series of networks. Calcium and phosphorus are utilized for insulin secretion and synthesis of high energy compounds. Adipose tissue store lipids and cholecalciferol, which, in turn, can influence calcium balance and energy expenditure. Hormones long-thought to solely modulate energy and mineral homeostasis may influence adipocytic function. Osteoblasts are a target of insulin action in bone. Moreover, endocrine mediators, such as osteocalcin, are synthesized in the skeleton but regulate carbohydrate disposal and insulin secretion. Finally, osteoblasts and adipocytes originate from the same mesenchymal progenitor. The mutual crosstalk between osteoblasts and adipocytes within the bone marrow microenvironment plays a crucial role in bone remodeling. In the present review we provide an overview of the reciprocal control between bone and energy metabolism and its clinical implications.

Effects of Electromagnetic Pulse on Bone Metabolism of Mice in vivo

Objective To study the effects of electromagnetic pulse （EMP） on bone metabolism of mice in vivo. Methods Twenty-four male BALB/c mice were divided into a control group and 2 experimental groups （n=8）. The whole-body of mice in experimental groups were exposed to 50 kV/m and 400kV/m EMP, 400 pulses daily for 7 consecutive days at 2 seconds intervals. Alkaline phosphotase （ALP） activity, serum calcium concentration and osteocalcin level and trabecular bone volume （BV/TV, %） were measured immediately after EMP exposure by biochemical, ELISA and morphological methods. Results The ALP activity, serum calcium concentration and osteocalcin level and BV/TV in experimental groups remained unchanged after EMP exposure. Conelusion Under our experimental conditions, EMP exposure cannot affect bone metabolism of mice in vivo.

Energy metabolism and the skeleton:Reciprocal interplay

The relation between bone remodelling and energy expenditure is an intriguing,and yet unexplained,challenge of the past ten years. In fact,it was only in the last few years that the skeleton was found to function,not only in its obvious roles of body support and protection,but also as an important part of the endocrine system. In particular,bone produces different hormones,like osteocalcin(OC),which influences energy expenditure in humans. The undercarboxylated form of OC has a reduced affinity for hydroxyapatite; hence it enters the systemic circulation more easily and exerts its metabolic functions for the proliferation of pancreatic β-cells,insulin secretion,sensitivity,and glucose tolerance. Leptin,a hormone synthesized by adipocytes,also has an effect on both bone remodelling and energy expenditure; in fact it inhibits appetite through hypothalamic influence and,in bone,stimulates osteoblastic differentiation and inhibits apoptosis. Leptin and serotonin exert opposite influences on bone mass accrual,but several features suggest that they might operate in the same pathway through a sympathetic tone. Serotonin,in fact,acts via two opposite pathways in controlling bone remodelling: central and peripheral. Serotonin product by the gastrointestinal tract(95%) augments bone formation by osteoblast,whereas brain-derived serotonin influences low bone mineral density and its decrease leads to an increase in boneresorption parameters. Finally,amylin(AMY) acts as a hormone that alters physiological responses related to feeding,and plays a role as a growth factor in bone. In vitro AMY stimulates the proliferation of osteoblasts,and osteoclast differentiation. Here we summarize the evidence that links energy expenditure and bone remodelling,with particular regard to humans.

Decarboxylated osteocalcin,a possible drug for type 2 diabetes,triggers glucose uptake in MG63 cells

BACKGROUND Uncarboxylated osteocalcin(GluOC)has been reported to improve glucose metabolism,prevent type 2 diabetes,and decrease the severity of obesity in mice with type 2 diabetes.GluOC can increase glucose uptake in a variety of cells.Glucose metabolism is the main source of energy for osteoblast proliferation and differentiation.We hypothesized that decarboxylated osteocalcin(dcOC),a kind of GluOC,can increase glucose uptake in MG63 cells(osteoblast-like osteosarcoma cells)and influence their proliferation and differentiation.AIM To investigate the effects of dcOC on glucose uptake in human osteoblast-like osteosarcoma cells and the possible signaling pathways involved.METHODS MG63 cells(human osteoblast-like osteosarcoma cells)were treated with dcOC(0,0.3,3,10,or 30 ng/mL)for 1 and 72 h,and glucose uptake was measured by flow cytometry.The effect of dcOC on cell proliferation was measured with a CCK-8 assay,and alkaline phosphatase(ALP)enzyme activity was measured.PI3K was inhibited with LY294002,and hypoxia-inducible factor 1 alpha(HIF-1α)was silenced with siRNA.Then,GPRC6A(G protein-coupled receptor family C group 6 subtype A),total Akt,phosphorylated Akt,HIF-1α,and glucose transporter 1(GLUT1)levels were measured by Western blot to elucidate the possible pathways by which dcOC modulates glucose uptake.RESULTS The glucose uptake of MG63 cells was significantly increased compared with that of the paired control cells after short-term(1 h)treatment with dcOC at different concentrations(0.3,3,and 10 ng/mL groups,P<0.01;30 ng/mL group,P<0.05).Glucose uptake of MG63 cells was significantly increased compared with that of the paired control cells after long-term(72 h)treatment with dcOC at different concentrations(0.3,3,and 10 ng/mL groups,P<0.01;30 ng/mL group,P<0.05).DcOC triggered Akt phosphorylation in a dose-dependent manner,and the most effective stimulatory concentration of dcOC for short-term(1 h)was 3 ng/mL(P<0.01).LY294002 abolished the dcOC-mediated(1 h)promotion of Akt phosphorylation and glucose uptake without affecting GLUT1 protein expression.Long-term dcOC stimulation triggered Akt phosphorylation and increased the protein levels of HIF-1α,GLUT1,and Runx2 in a dose-dependent manner.Inhibition of HIF-1αwith siRNA abolished the dcOC-mediated glucose uptake and substantially decreased GLUT1 protein expression.DcOC interven-tion promoted cell proliferation in a time-and dose-dependent manner as determined by the CCK-8 assay.Treatment with both 3 ng/mL and 10 ng/mL dcOC affected the ALP activity in MG63 cells after 72 h(P<0.01).CONCLUSION Short-and long-term dcOC treatment can increase glucose uptake and affect proliferation and ALP activity in MG63 cells.This effect may occur through the PI3K/Akt,HIF-1α,and GLUT1 signaling factors.

Study of vascular smooth muscle cell calcification induced by hyperphosphate and intervented by phosphonoformic acid

Objective： To evaluate the effects of different concentrations of phosphate on calcium deposition and osteocalcin level in cultured bovine aortic smooth muscle cell, investigate the mechanism of hyperphosphatemia to evoke calcification of vascular smooth muscle cell and observe the effects of phosphonoformic acid（PFA） in different concentrations on vascular calcification. Methods： The bovine aortic smooth muscle cells （BASMC） were cultured.Calcium deposition and the expression of osteocalcin of BASMC in different concentrations of phosphate （1.5 mmol/L and 2.0 mmol/L） and PFA were determined by o-cresolphthalein complexone and radioimmunity methods, respectively. Osteocalcin mRNA expressions were determined by RT-PCR. Results： After six or nine days of BASMC cultured, the calcium deposition in Pi 2.0 mmol/L group was more than that in Pi 1.5 mmol/L group[（77.187 ± 11.692） lag/（mg · protein） vs（25.768 ± 1.750）lag/（mg · protein）, P 〈 0.01 and（125.399 ± 16.677）lag/（mg · protein） vs（29.046 ± 2.635）lag/（mg · protein）, P 〈 0.01 respectively]. The calcium deposition was dependent on time and dosage of phosphate treatment. After 72 h culture the osteocalcin in Pi 2.0 mmol/L group was more than that in Pi 1.5 mmol/L grouplin supematant,（1.503 ± 10^-2 ± 2.601 × 10^-3）ng/（ lag o protein） vs（2.981 × 10-3 ± 8.382 × 10-34）ng/（ lag · protein）, P 〈 0.001], the same was found in osteocalcin mRNA expression[OC/GAPDH, （1.906 ± 0.132） vs（0.748 ± 0.037）, P〈 0.001]. Compared to Pi 1.5mmol/L group,bovine smooth muscle cells（BSMC） cultured in media containing Pi 2.0 mmol/L phosphate levels increased calcium deposition[On day 6,（77.187 ± 11.692） la g/（mg · protein） vs （25.768 ±1.750） la g/（mg · protein）, P 〈 0.001]. Elevated phosphate treatment of BSMCs also enhanced the expression of the osteoblastic differentiation marker osteocalcin[On day 3, Pi 2.0 mmol/L group vs Pi 1.5mmol/L group,（1.503 × 10^-2 ± 2.601 × 10^-3 ）ng/（ lag · protein） vs（2.981× 10^-3 ± 8.382 × 10^-4）ng/（ μg · protein）, P 〈 0.001]. PFA decreased ciacium deposition and osteocalcin expression statistically[Pi 2.0 mmol/L±PFA1.0 mmol/L group vs Pi 2.0mmol/L group, ciacium deposition, （37.729 ± 5.899） lμg/（mg · protein） vs （77.187 ± 11.692）μg/（mg ·protein）, P 〈 0.001]; Osteocalcin in supernatant, （4.529 ± 10^-3 ± 1.250 × 10^-3）ng/（ μ g · protein） vs（1.503 × 10^-2 ± 2.601 × 10^-3） ng/（ μg · protein）, P〈 0.001; osteocalcin mRNA expression, OC/GAPDH, （0.642 ± 0.092） v s （1.89 ± 0.165）, P 〈 0.01]. Conclusion： Hyperphosphate may directly promote calcium deposition and the osteocalcin expression of B ASMCs. It may be a new explanation for the phenomenon of vascular calcification in hyperphosphatemic conditions. Hyperphosphatemia is an independent factor to stimulate vascular calcification. PFA can inhibit calcium deposition and osteocalcin expression induced by elevated phosphate.PFA may be a new medicine to treat vascular calcification induced by elevated phosphate.

Relationship between the Components of the Metabolic Syndrome and Measures of Bone Mineral Density in Post-Menopausal Women

Aim: To examine the association between individual components of metabolic syndrome (MetS) and bone mineral density (BMD) among postmenopausal women. Methods: A total of 177 postmenopausal women participated in a cross-sectional study. They were interviewed to collect anthropometric and demographic characteristics. BMD was measured and biochemical parameters were estimated in fasting blood samples. Univariate and multivariate analyses were used to examine the association between individual components of MetS and BMD. Results: Among 177 postmenopausal women, 116 (66%) had MetS. Women with MetS had significantly higher mean values of BMD and T scores at the total hip (P < 0.05) compared to women without MetS, which disappeared after adjustment for body weight, but not for age (P < 0.05). Features of the MetS other than waist circumference were not significantly related to BMD values at the three skeletal sites, except for diastolic blood pressure association with BMD at the femoral neck (r = 0.150, P < 0.05). BMD at the total hip was also positively associated with both of triglycerides (r = 0.157, P < 0.05) and fasting blood glucose (r = 0.193, P < 0.01). To identify the independent factors affecting the BMD at the 3 skeletal sites according to metabolic states, stepwise multiple linear regression analysis was performed. Conclusions: Body weight and osteocalcin were more strongly associated with bone mass than any other component of MetS in postmenopausal women. However, further studies seem to be needed to confirm their observation.

Negative association between trunk fat, insulin resistance and skeleton in obese women

AIM: To evaluate the potential interference of trunk fat (TF) mass on metabolic and skeletal metabolism. METHODS: In this cross-sectional study, 340 obese women (mean age: 44.8 ± 14 years; body mass index: 36.0 ± 5.9 kg/m 2 ) were included. Patients were evaluated for serum vitamin D, osteocalcin (OSCA), inflammatory markers, lipids, glucose and insulin (homeostasis model assessment of insulin resistance, HOMA-IR) levels, and hormones profile. Moreover, all patients underwent measurements of bone mineral density (BMD;at lumbar and hip site) and body composition (lean mass, total and trunk fat mass) by dual-energy X-ray absorptiometry. RESULTS: Data showed that: (1) high TF mass was inversely correlated with low BMD both at lumbar (P < 0.001) and hip (P < 0.01) sites and with serum vitamin D (P < 0.0005), OSCA (P < 0.0001) and insulin-like growth factor-1 (IGF-1; P < 0.0001) levels; (2) a positive correlation was found between TF and HOMA-IR (P < 0.01), fibrinogen (P < 0.0001) and erythrocyte sedimentation rate (P < 0.0001); (3) vitamin D levels were directly correlated with IGF-1 (P < 0.0005), lumbar (P < 0.006) and hip (P < 0.01) BMD; and (4) inversely with HOMA-IR (P < 0.001) and fibrinogen (P < 0.0005). Multivariate analysis demonstrated that only vitamin D was independent of TF variable. CONCLUSION: In obese women, TF negatively correlates with BMD independently from vitamin D levels. Reduced IGF-1 and increased inflammatory markers might be some important determinants that account for this relationship.

Arterial calcification: Finger-pointing at resident and circulating stem cells

The term "Stammzelle"(stem cells) originally appeared in 1868 in the works of Ernst Haeckel who used it to describe the ancestor unicellular organism from which he presumed all multicellular organisms evolved. Since then stem cells have been studied in a wide spectrum of normal and pathological conditions; it is remarkable to note that ectopic arterial calcification was considered a passive deposit of calcium since its original discovering in 1877; in the last decades, resident and circulating stem cells were imaged to drive arterial calcification through chondro-osteogenic differentiation thus opening the idea that an active mechanism could be at the basis of the process that clinically shows a Janus effect: calcifications either lead to the stabilization or rupture of the atherosclerotic plaques. A review of the literature underlines that 130 years after stem cell discovery, antigenic markers of stem cells are still debated and the identification of the osteoprogenitor phenotype is even more elusive due to tissue degradation occurring at processing and manipulation. It is necessary to find a consensus to perform comparable studies that implies phenotypic recognition of stem cells antigens. A hypothesis is based on the singular morphology and amitotic mechanism of division of osteoclasts: it constitutes the opening to a new approach on osteoprogenitors markers and recognition. Our aim was to highlight all the present evidences of the active calcification process, summarize the different cellular types involved, and discuss a novel approach to discover osteoprogenitor phenotypes in arterial wall.

Effects of β-TCP Ceramics on Osteoblast Cellular Proliferating,Mineralization and Osteocalcin Expression

After co-cultrured osteoblast with fl-TCP ceramics, the cellular proliferating, mineralization and osteocalcin expression were studied. MTT assay showed that fl-TCP ceramics had no affect on cellular proliferating. Laser scanning confocal detection showed that fl-TCP ceramics could increase the mineralization level of osteoblast. Furthermore, RT-PCR showed that fl-TCP could increase the expression level of osteocalcin. Those results indicate β-TCP ceramics had perfect biocompatibility and increased the mineralization of osteoblast to accelerate osteogenesis by means of affecting the expression of genes involving in osteogeneticprocess.

Differentiation of Osteoblast in vitro Is Regulated by Progesterone

The regulation of cellular differentiation by progesterone in fetal rat calvarial osteoblasts was investigated. Our results showed that cells cultured in the presence of progesterone had a 7% increase in the alkaline phosphatase activity when compared to untreated cells. The concentration of osteocalcin in the conditioned medium from progesterone treated osteoblasts was 28% higher than that of untreated controls. In addition,administration of progesterone significantly enhanced the number and area of bone nodules. In conclusion, progesterone stimulates the differentiation of fetal rat calvarial osteoblastic cells in vitro

RU486 Reversal of Cortisol Repression of 1,25-Dihydroxyvitamin D₃Induction of the Human Osteocalcin Promoter

In conditions of corticosteroid excess, such as Cushing’s syndrome, a reduction in serum osteocalcin is observed and bone loss occurs. The human osteocalcin gene is induced by 1,25-dihydroxyvitamin D3 derivatives and repressed by glucocorticoids. In this paper we show that cortisol, a natural glucocorticoid, represses both basal and vitamin D induced activity of the human osteocalcin promoter. Furthermore, we address the specific question as to whether the anti-progestin anti-glucocorticoid RU486 is able to antagonize the inhibitory effect of cortisol on osteocalcin gene expression. We show that RU486 has agonist activity alone, in that it is able to repress the basal promoter activity of the osteocalcin gene and antagonist activity, reversing incompletely the cortisol mediated repression of 1,25-dihydroxyvitamin D3 induction.

Laboratory Findings in Twins Suffering from Autism—Case Report

Autism refers a wide range of conditions and the most prominent are challenges with social skills, repetitive behaviors, speech disturbances. By now, it is known that there is no one type of autism, but there are many of them which are caused by genetic factors and environmental influences. Sometimes autism is called autism spectrum disorders which reflect the wide variation in challenges and strengths possessed by each person with autism (e.g., gastrointestinal disorders, seizures, sleep disturbances, attention deficit and hyperactivity disorders, anxiety etc.). Around one third of all people with autism remain nonverbal and have an intellectual disability. This condition can be detected as early as 18 months, but the most obvious signs tend to appear between 2 and 3 years of age. Only early intervention in those patients can improve outcomes. There are no many articles published regarding laboratory findings in autism patients. By performing holistic approach in patient healing, we also face autistic children in everyday practice. In this paper we represented case of twins suffering from autism. We found that both of them had elevation of free triiodothyronine, oxytocin, osteocalcin, and basophils. We found that vitamin D level in both twins was lower than recommended. Further research in this field is necessary to reveal whether these findings are causal factors for autism or are an effect of the disease.

Relationship between serum osteocalcin level and glucose and lipid metabolism in patients with type 2 diabetes mellitus complicated with nonalcoholic fatty liver disease

Objective: To study the relationship between serum osteocalcin level and glucose and lipid metabolism in patients with type 2 diabetes mellitus complicated with nonalcoholic fatty liver disease. Methods: The clinical data of 180 type 2 diabetes mellitus patients in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, China from February 2017 to January 2018 were retrospectively analyzed, including 90 cases of nonalcoholic fatty liver disease patients (group A) and 90 cases of nonalcoholic fatty liver disease-free patients (group B), meanwhile another 100 healthy subjects were selected as the control group. Then various indexes were compared between groups, including serum osteocalcin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), prothrombin activity (PTA), fasting blood glucose (FBG), glycated hemoglobin (HbA1c), triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), fasting insulin (FINS), fasting C peptide (FCP), HOMA insulin resistance index (HOMA-IR), HOMA β-cell function (HOMA-β). Results: The serum osteocalcin and PTA in group A were significantly lower than those in group B and the control group (P<0.05). ALT, AST, and ALP were significantly higher than those in group B and the control group (P<0.05). The FBG and HbA1c in group A were significantly higher than those in group B and the control group (P<0.05). The TG, TC, LDL, and HDL of group A and group B were significantly higher than those of the control group (P<0.05). The FINS, FCP, and HOMA-IR in group A were significantly higher than those in group B and the control group (P<0.05). HOMA-βwas significantly lower than group B and the control group (P<0.05). Pearson correlation analysis showed that the serum osteocalcin was not correlated with ALT, AST, ALP, PTA, HbA1c, TG, TC, LDL, HDL and FINS (P>0.05), but negatively correlated with FBG and HOMA-IR (P<0.05), and positively correlated with FCP and HOMA-β (P<0.05). With serum osteocalcin as the dependent variable, and ALT, AST, ALP, PTA, FBG, HbA1c, TG, TC, LDL, HDL, FINS, FCP, HOMA-IR and HOMA-β as independent variables, multiple stepwise regression analysis showed that the FBG, HOMA-IR and HOMA-β were independent risk factors for osteocalcin. Conclusions: Patients with type 2 diabetes mellitus complicated with nonalcoholic fatty liver disease have lower serum osteocalcin level, which is susceptible to FBG, HOMA-IR, HOMA-β, and other factors.