Multi-element determination in cancellous bone of human femoral head by PIXE

Proton Induced X-ray Emission (PIXE) method is used for the determination of elemental concentrations in cancellous bone of human femoral head from five autopsies and seven patients with femoral neck broken. The specimen preparation and experimental procedure are described in detail. Using the t test, the results show that the concentrations of P, Ca, Fe, Cu, Sr in control group are higher than those in patient group, but the concentrations of S, K, Zn, Mn are not significantly different. The physiological functions of metallic elements in human bone are also discussed.     

Comparative proteomics analysis of microvesicles in human serum for the evaluation of osteoporosis

Osteoporosis is an emerging health issue worldwide. Due to the decrease of bone mineral density and the deterioration of skeletal microarchitecture, osteoporosis could lead to increased bone fragility and higher fracture risk. Since lack of specific symptoms, novel serum proteomic indicators are urgently needed for the evaluation of osteoporosis. Microvesicles (MVs) are important messengers widely present in body fluids and have emerged as novel targets for the diagnosis of multiple diseases. In this study, MVs were successfully isolated from human serum and comprehensively characterized. Comparative proteomics analysis revealed differential MVs protein profiling in normal subjects, osteopenia patients, and osteoporosis patients. In total, about 200 proteins were identified and quantified from serum MVs, among which 19 proteins were upregulated (fold change >2) and five proteins were downregulated (fold change <0.5) in osteopenia group and osteoporosis group when compared with the normal group. Three protein candidates were selected for initial verification, including Vinculin, Filamin A, and Profilin 1. Profilin 1 was further pre‐validated in an independent sample set, which could differentiate osteoporosis group from osteopenia group and normal group (p < 0.05). Our data collectively demonstrate that serum MVs proteome can be valuable indicators for the evaluation and diagnostics of bone loss disease.     

健骨胶囊中微量元素对实验大鼠股骨的影响

通过对健骨胶囊及实验大鼠股骨中的微量元素铜、锌、铁、锰的分析，阐述了健骨胶囊中微量元素对实验大鼠股骨中的微量元素铜、锌、铁、锰的影响以及骨质疏松症的改善，具有重要的营养学、药理功效学作用。     

骨质疏松与微量元素

骨质疏松与某些矿物质的饮食摄取不足、过量或不平衡有关，但微量元素在骨质疏松的病理、诊断和治疗中也起着十分重要的作用。适量的Ｂ、Ｆ、Ｓｉ、Ｍｎ、Ｃｕ、Ｚｎ、Ｇｅ、Ｓｒ和Ｖ、Ｉ、Ｍｏ对骨骼的发育和代谢是必需的，但Ｐｂ、Ｃｄ、Ａｌ则可能是骨质疏松的危险因子。高尿锌可被用作骨质疏松的生物化学标记物，锶可代替钙用作骨矿化的标记物。临床实践表明，氟可明显增加骨质疏松患者的长轴骨密度和显著降低脊椎骨新骨折的发生     

Application of neutron activation analysis to the study of element concentration and exchange in fossil bones

It has long been known that a variety of elements are introduced into voids in the structure of bone during fossilization. Visual examination of the cross-section of many bones in the early stages of fossilization reveal a staining which is most intense near the outer surface. This suggests that concentration gradients must exist for elements entering the bone. To investigate this possibility quantitatively, we have determined elemental concentrations in such bones as a function of depth below the outer surface. Using a SLOWPOKE reactor, we have analysed a number of samples of bone taken from secondary deposits of river sediments in the Old Crow region of the Yukon Territory, Canada. Our preliminary work indicates the decrease in concentration with depth into the cortex for previously recognized post-mortal elements such as U, F, Ba, Mn and Fe. Our measurements show that V, Sc, and Co also vary in this way and can be included in this group. As the hollow central shafts of bones are approached, concentrations increase again. Bones found in the Old Crow region display a variety of surface staining ranging from almost white through red to brown and black. There is a strong correlation between Fe and less positive relations between F and Ba, and staining. Highest Mn concentrations occur at the surfaces of light-coloured bones in which the staining is restricted to the outer 1 mm. Because of the wide range of concentrations which exist amont post-mortal elements in these bones, correlations among these elements can be studied. Manganese and barium, for example, are correlated within each bone, but the correlation patterns differ from bone to bone. These relationships must reflect the nature of the chemical processes associated with the deposition of the elements within the bones. A comparison of these patterns show that some of the bones have been exposed to more than one set of environmental conditions. This data thus provides a way of studying these processes in bones in which the introduction of elements from the environment has not completely and uniformly filled all the available voids.     

毛冬青总皂苷对AHNP诱导肝损伤大鼠骨中7种微量元素的影响

目的探讨了毛冬青总皂苷对AHNP诱导肝损伤大鼠骨中微量元素的影响。方法应用逆行胰胆管术注射牛磺胆酸钠（TAC）复制急性出血坏死性胰腺炎（AHNP）大鼠实验动物模型,进行了毛冬青总皂苷（高、低剂量）的治疗。观察了5组（假手术对照组、AHNP模型组、毛冬青总皂苷高剂量组、毛冬青总皂苷低剂量组、尼尔雌醇组、）大鼠股骨中Cu、Mn、Fe、Zn、Pb、Mg和ca等微量元素的变化。结果统计学结果显示与AHNP模型组比较,毛冬青总皂苷能提高AHNP大鼠骨质中cu、Fe、Mn、Mg、Ca等元素的含量,尼尔雌醇组中的zn含量提高,毛冬青总皂苷组中的zn含量反而下降;尼尔雌醇组与毛冬青总皂苷组中Pb含量下降明显。结论说明毛冬青总皂苷的治疗作用并不是单一的,可以认为中药毛冬青总皂苷是通过发挥对人体整体的调节作用来调节骨骼中微量元素的变化,改变骨质的代谢情况,促进骨密度的提高。     

Trace elements in human bone determined by neutron activation analysis

There is an evidence that some of the essential trace elements are crucial determinants of bone health. Excess or deficiency of these elements has a role in the development of bone diseases, therefore research on trace elements in bone is very important. Iliac crest bone biopsies were optioned from twelve persons undergoing orthopedic surgery due to any reason than osteoporosis. Cortical and trabecular parts were separated, and blood and fats were removed. Up to 30 minor and trace elements were determined in these samples by instrumental neutron activation analysis and other techniques and their relations were discussed.     

Anti-Osteoporotic Effects of n-trans-Hibiscusamide and Its Derivative Alleviate Ovariectomy-Induced Bone Loss in Mice by Regulating RANKL-Induced Signaling

Osteoporosis is characterized by the deterioration of bone structures and decreased bone mass, leading to an increased risk of fracture. Estrogen deficiency in postmenopausal women and aging are major factors of osteoporosis and are some of the reasons for reduced quality of life. In this study, we investigated the effects of n-trans-hibiscusamide (NHA) and its derivative 4-O-(E)-feruloyl-N-(E)-hibiscusamide (HAD) on receptor activator of nuclear factor kappa-Β (NF-κB) ligand (RANKL)-induced osteoclast differentiation and an ovariectomized osteoporosis mouse model. NHA and HAD significantly inhibited the differentiation of osteoclasts from bone marrow-derived macrophages (BMMs) and the expression of osteoclast differentiation-related genes. At the molecular level, NHA and HAD significantly downregulated the phosphorylation of mitogen-activated protein kinase (MAPK) signaling molecules. However, Akt and NF-κB phosphorylation was inhibited only after NHA or HAD treatment. In the ovariectomy (OVX)-induced osteoporosis model, both NHA and HAD effectively improved trabecular bone structure. C-terminal telopeptide (CTX), a bone resorption marker, and RANKL, an osteoclast stimulation factor, were significantly reduced by NHA and HAD. The tartrate-resistant acid phosphatase (TRAP)-stained area, which indicates the osteoclast area, was also decreased by these compounds. These results show the potential of NHA and HAD as therapeutic agents for osteoporosis.     

骨的微量元素代谢

雌激素可通过影响骨骼细胞的新陈代谢而促进微量元素在骨骼中的储存,微量元素代谢与骨质疏松的形成发展有密切关系。近年来不少学者对一些必需微量元素如硼(5B)、氟(9F)、钒(23V)、锰(25Mn)、铁(26Fe)、钴(27Co)、铜(29Cu)、锌(30Zn)、硒(34Se)、锶(38Sr)等和一些非必需微量元素如铝(13Al)、镓(31Ga)、锗(32Ge)、镉(48Cd)、铅(82Pb)等与骨质疏松关系的研究取得了令人瞩目的成果。     

基于MRI的绝经后女性腰椎退行性病变程度与骨质疏松的相关性研究

本研究探讨基于磁共振成像(MRI)的绝经后女性腰椎退行性病变程度与骨质疏松的关系。选取经骨密度检查确诊的骨质疏松绝经后女性患者50例为骨质疏松组、非骨质疏松症绝经后妇女50例作为对照组,两组妇女均接受MRI检查。结果显示,骨质疏松组的L1/L2、L2/L3、L3/L4、L4/L5、L1~S1椎间盘Pfirrmann分级评分均高于对照组,差异均具有统计学意义(P<0.05);骨质疏松组患者的腰椎L1/L2、L2/L3、L3/L4、L4/L5、L1~S1之间的BMD与椎间盘Pfirrmann分级评分呈显著的正相关关系(P<0.05);多元线性回归模型结果显示:绝经时间、年龄、腰椎BMD值与患者椎间盘Pfirrmann分级评分呈显著的正相关关系(P<0.05)。绝经后女性采用MRI检查评估的腰椎退行性病变程度Pfirrmann分级评分与骨质疏松程度有关,同时与患者的年龄、绝经时间长度有关。     

血清外泌体的蛋白质组分析及其在骨质疏松中的应用

外泌体是细胞分泌的微小具膜囊泡，作为重要媒介参与细胞间的信号传递，已在疾病的诊断和治疗中发挥独特作用。骨质疏松症是全身性骨代谢疾病，容易引起骨密度减少并导致骨折，在老年人群中发病率很高，目前急需发展特异性体液诊断技术。本论文采用超速离心的方法，对血清中的外泌体进行了分离富集和表征，并采用液相色谱-质谱进行了外泌体蛋白质组学分析，共鉴定到了179个外泌体蛋白质，主要参与防御响应和免疫应答等生物过程。针对来自正常对照组、骨量减少组和骨质疏松组的血清样本，分离富集其中的外泌体，通过免标记定量蛋白质组学分析，分别鉴定到188、224和185个蛋白质。定量分析显示17个蛋白质的表达量在骨质疏松组和骨量减少组有显著改变（p<0.05），包括Integrin β 3、Integrin α 2 β 1、Talin 1和Gelsolin等，说明人体骨质在衰变过程中发生了系统性变化，并体现在血清外泌体中。该研究可为骨质疏松研究提供潜在的分子标志物，有助于阐明其病变机制。     

Chitooligosaccharides prevent osteopenia by promoting bone formation and suppressing bone resorption in ovariectomised rats: possible involvement of COX-2

Chitooligosaccharides (CHOS) added in diet have been found as potent calcium fortifiers in conditions of Ca²⁺ deficiency such as osteoporosis. In this study, we found that pharmaceutical intervention using CHOS prevented ovariectomy (OVX)-induced bone mineral density loss and the deterioration of trabecular microarchitecture in a dose-dependent manner (p < 0.05 or 0.01). CHOS (125, 250 mg/kg) suppressed the serum levels of bone resorption biomarkers CTx and TRACP5b induced by OVX (p < 0.05), but increased the levels of osteogenic markers ALP and OC by 11.3–11.6% and 10.7–15.2% of OVX group (p < 0.05), suggesting the exact pharmacological action of CHOS in the control of osteoporosis which may be the result of both promoting bone formation and suppressing bone resorption. Bone turnover-modulating effects of CHOS appear related to their anti-inflammatory capacity to down-regulate mRNA and protein expression of COX-2 (17.2–32.2% and 16.4–21.9% of OVX group, p < 0.05 or 0.01), a key mediator linking between inflammation and osteoporosis.     

Comparison of elemental composition of hair between osteoporotic and normal women by instrumental neutron activation analysis

Instrumental neutron activation analysis (INAA) was carried out on hair samples from a group of twenty patients undergoing a study of osteoporosis. Half of these were judged normal. Fourteen elements were measured but only calcium was found to have a correlation with the disease state. Calcium levels in hair were significantly lower (99% confidence level) in patients with osteoporosis. Conversely, calcium levels in the blood of osteoporotic patients were significantly higher (95% confidence level) than those in normals. Though the group studied was small it is felt that the levels of calcium in hair may be of value in diagnosing osteoporosis.     

Trace elements distribution and post-mortem intake in human bones from Middle Age by total reflection X-ray fluorescence

The purpose of the present work is to investigate the suitability of TXRF technique to study the distribution of trace elements along human bones of the 13th century, to conclude about environmental conditions and dietary habits of old populations and to study the uptake of some elements from the surrounding soil.In this work, we used TXRF to quantify and to make profiles of the elements through long bones. Two femur bones, one from a man and another from a woman, buried in the same grave were cross-sectioned in four different points at a distance of 1 cm. Microsamples of each section were taken at a distance of 1 mm from each other.Quantitative analysis was performed for Ca, Mn, Fe, Cu, Zn, Sr, Ba and Pb. Very high concentrations of Mn and Fe were obtained in the whole analysed samples, reaching values higher than 2% in some samples of trabecular tissue, very much alike to the concentrations in the burial soil. A sharp decrease for both elements was observed in cortical tissue. Zn and Sr present steady concentration levels in both kinds of bone tissues. Pb and Cu show very low concentrations in the inner tissue of cortical bone. However, these concentrations increase in the regions in contact to trabecular tissue and external surface in contact with the soil, where high levels of both elements were found. We suggest that contamination from the surrounding soil exists for Mn and Fe in the whole bone tissue. Pb can be both from post-mortem and ante-mortem origin. Inner compact tissue might represent in vivo accumulation and trabecular one corresponds to uptake during burial. The steady levels of Sr and Zn together with soil concentration lower levels for these elements may allow us to conclude that they are originated from in vivo incorporation in the hydroxyapatite bone matrix.     

New Generation of Meso and Antiprogestins (SPRMs) into the Osteoporosis Approach

Receptor activator of nuclear factor κB (RANK) and its ligand (RANKL) play key roles in bone metabolism and the immune system. The RANK/RANKL complex has also been shown to be critical in the formation of mammary epithelia cells. The female hormones estradiol and progesterone closely control the action of RANKL with RANK. Blood concentration of these sex hormones in the postmenopausal period leads to an increase in RANK/RANKL signaling and are a major cause of women’s osteoporosis, characterized by altered bone mineralization. Knowledge of the biochemical relationships between hormones and RANK/RANKL signaling provides the opportunity to design novel therapeutic agents to inhibit bone loss, based on the anti-RANKL treatment and inhibition of its interaction with the RANK receptor. The new generation of both anti- and mesoprogestins that inhibit the NF-κB-cyclin D1 axis and blocks the binding of RANKL to RANK can be considered as a potential source of new RANK receptor ligands with anti-RANKL function, which may provide a new perspective into osteoporosis treatment itself as well as limit the osteoporosis rise during breast cancer metastasis to the bone.     

The determination of strontium in human blood serum and packed blood cells by radiochemical neutron activation analysis

Strontium is a trace element whose distribution and metabolism are similar to those of calcium, its close chemical analogue. The element received scientific attention because of its potential beneficial effects in the treatment of osteoporosis andosteolytic metastatic bone lesions but also because of the possible problems associated with the retention of⁹⁰Sr from radioactive fall-out. In continuation of a series of experiments intended to establish reliable reference values for trace elements in human blood serum and packed blood cells, we set up a project to determine also strontium. The following values were obtained (mean±standard deviation): 22.2±4.8 ng/ml (serum) and 2.85±1.11 ng/g wet weight (packed blood cells).     

大豆异黄酮对骨质疏松模型大鼠骨量及微观结构的影响

为研究大豆异黄酮（SI）对骨质疏松模型大鼠骨量及微观结构的影响,将46只雌性SD大鼠双侧摘除卵巢,术后一周按血清总胆固醇水平随机分为5组,分别给予雌激素,低、中、高剂量SI干预并设正常对照组,实验周期为12周,灌胃给药,每周称质量一次,分别在去卵巢、给药4、8周处死时抽取尾静脉血,实验结束后分别测定血清碱性磷酸酶（AKP）活性及骨密度等指标。结果表明,高剂量SI干预对维持大鼠骨密度的作用与雌激素相似,去卵巢后干预SI可提高碱性磷酸酶（AKP）活性并能缓解因去卵巢造成的骨丢失,全视野下骨形态学以雌激素组、高剂量SI干预组与正常组结构最为接近。提示大豆异黄酮对骨质疏松模型大鼠的骨量及微观结构存在一定影响,中、高剂量干预可使骨质疏松模型大鼠血清AKP酶活性增加、逆转因去势造成的骨密度下降,考虑到植物雌激素与哺乳动物的雌激素受体（ER）结合能力低下,采用SI在临床开展干预的剂量与远期效果尚待进一步论证。     

氟化物对幼年大鼠骨矿物质和血微量元素的影响及其相关性研究

为探讨氟化钠对幼年大鼠血清4种微量元素和骨矿物质含量的影响,以及大鼠骨矿物质和血清微量元素的相关性,将80只2月龄SPF级SO大鼠,雌雄各半,随机分成8组：对照（幼年CS、成年AS）组和用药组（幼年高氟组CHS、幼年低氟组CLS、成年高氟组AHS、成年低氟组ALS、幼年长期高氟组HS、幼年长期低氟组15）。对照组灌胃生理盐水,用药组分别按相应时间给予不同剂量的NaF灌胃,测定了大鼠尺骨和血清中的Ca、P、Fe、Zn含量。结果表明,（1）血清微量元素：①与cs组相比,CHS组和CLS组的磷（P）分别增加100．0％和193．8％（P〈0．05）。②与AS组相比,HS组的Ca、Zn分别下降了20．5％和40．0％,而P则增加了74．0％（P〈0．05）;Ls组的Zn下降了33．3％（P〈0．05）。与HS组比较,CHS组的Ca增加22．3％。③与LS组比较,CLS组的P和Zn明显增加49．2％和28．5％（P〈0．05）。（2）骨矿物质：①与As组相比,HS组的Ca分别下降了23．9％（P〈0．05）,Zn则增加了36．1％（P〈0．05）。②与HS组相比,CHS组的Ca和Fe分别增加了42．3％和36．05％,Zn下降了38．6％（P〈0．05）。（3）骨矿物质和血清微量元素的相关性：骨Ca和血Ca、骨P和血P、骨Zn和血Zn、骨Fe和血Fe的相关系数分别为0．435、0．347、0．136和0．059（P〉0．05）。提示长期使用氟化钠,可以导致幼年大鼠血中微量元素代谢紊乱,骨矿物含量降低,但未确定大鼠骨矿物含量和血微量元素具有相关性。     

藏药牦牛肝提取物中矿物质元素特征及意义

采用220FS原子吸收光谱仪测定了传统藏药牦牛肝提取物中的K、Na、Ca、Mg、Fe、Cu、Zn、Mn等8种矿物质元素含量.结果显示,藏药牦牛肝提取物中含有丰富的矿物质元素,对儿童和青少年骨骼的生长发育,中老年骨质疏松症预防有着极为重要的营养学和治疗意义,开发前景广阔.     

利骨素抗去卵巢大鼠骨矿物元素代谢紊乱的作用机制

通过测定骨元素含量和血清相关激素及白细胞介素-6水平,探讨了利骨素对去卵巢大鼠骨元素代谢的影响及其机制。将健康4月龄雌性SD大鼠24只随机分成3组:A组,假去卵巢组;B组,去卵巢组;C组,去卵巢+利骨素组。结果表明,去卵巢大鼠骨Ca、Mg、S、Mn、Zn等含量显著降低,P含量显著增加;应用利骨素可使去卵巢大鼠降低的骨Ca、Mg、S、Mn、Zn等含量显著回升,P含量显著回降。血清相关激素:去卵巢组较假去卵巢组大鼠,血清雌二醇(E2)、孕酮(P)、促甲状腺激素(TSH)、甲状腺素(T4)、降钙素(CT)、皮质醇(Cortisol)、生长素(GH)等含量降低,卵泡刺激素(FSH)、黄体生成素(LH)、白细胞介素-6(IL-6)等含量升高;去卵巢+利骨素组较去卵巢组,血清P,GH,Cortisol,TSH,T4等含量回升,LH含量回降。提示利骨素改变去卵巢大鼠体内相关激素水平是其纠正去卵巢大鼠骨元素代谢紊乱作用的重要机制。