Stimulation of bone resorption and inhibition of bone formation in vitro by human tumour necrosis factors

When leukocytes are exposed to mitogens or antigens in vitro, they release bone-resorbing activity into the culture supernatants which can be detected by bioassay. Like many lymphocyte-monocyte products, this activity has been difficult to purify because of its low abundance in activated leukocyte cultures and the unwieldy bioassay required to detect biological activity. Partially purified preparations of this activity inhibit bone collagen synthesis in organ cultures of fetal rat calvariae. Recent data suggest that both activated lymphocytes and monocytes release factors which could contribute to this activity. Recently, monocyte-derived tumour necrosis factor alpha (TNF-alpha) and lymphocyte-derived tumour necrosis factor beta (TNF-beta) (previously called lymphotoxin), two multifunctional cytokines which have similar cytotoxic effects on neoplastic cell lines, have been purified to homogeneity and their complementary DNAs cloned and expressed in Escherichia coli. As both of these cytokines are likely to be present in activated leukocyte supernatants, we tested purified recombinant preparations for their effects on bone resorption and bone collagen synthesis in vitro, and report here that both cytokines at 10(-7) to 10(-9) M caused osteoclastic bone resorption and inhibited bone collagen synthesis. These data suggest that at least part of the bone-resorbing activity present in activated leukocyte culture supernatants may be due to these cytokines.     

Expression of human adenosine deaminase in murine haematopoietic progenitor cells following retroviral transfer

Adenosine deaminase (ADA) deficiency, an autosomal recessive inborn error of metabolism, leads to severe combined immune deficiency in man. This enzyme, although constitutively expressed in most tissues, is expressed at high level in immature T cells, and study of the pathophysiology of the disorder indicates that increased deoxyadenosine or altered methylation capacity have toxic effects on T-cell maturation. Although bone marrow transplantation can correct the immune deficiency, this therapy is associated with graft-versus-host disease and incomplete immune restoration, and so our laboratory and others have sought to develop a method of gene replacement as a possible treatment for the disease. Moreover, characterization of the complementary DNA of the human ADA gene and some of its mutants makes it possible to design gene transfer strategies. We have now subcloned a human adenosine deaminase cDNA into the retrovirus shuttle vector pZIP-SV(B), and in this way have isolated a cell line, 4.2T, which produces high titres of replication-defective retrovirus which have been used to transfer the gene for human ADA to mouse bone marrow cells. Transfer and expression of the neomycin-resistance gene (neo) and the ADA gene in murine bone marrow colony-forming units (CFU) was demonstrated by in vitro colony formation in the presence of the antibiotic G418 or 9-xylofuranosyladenine plus deoxycoformycin, respectively. Isoenzyme analysis also showed human ADA expression in the cultured mouse bone marrow.     

辽宁北票龙骨组织中类似血细胞结构的研究

在我国辽宁北票下白垩统义县组下部地层中出土的北票龙(Beipiaosaurus inexpectus)化石由部分椎骨和肢骨碎片构成,其中保存有血管和类似血细胞的结构。在肢骨周围有褐色细丝状的皮肤衍生物的印痕。细丝状皮肤衍生物无分枝,大约0.1mm宽,50～60mm长。在光学显微镜下观察到其肱骨的纵切面上有褐色血管结构,一些管腔中有褐色小圆球,它们沿着管腔分布。这些小圆球的形状和大小显示它们有可能是恐龙血细胞的遗迹。另外,图像分析法结果显示北票龙骨组织有较高的血管密度,表示它具有较高的新陈代谢率。从骨组织特征及作为体表隔热层的细丝状皮肤衍生物的存在来看,北票龙可能为恒温动物。     

海藻酸盐复合凝胶在骨组织工程中的研究进展

骨组织工程支架是以修复缺损的骨并恢复其功能为目的而开发出的人工支架。海藻酸盐是世界上含量最丰富的海洋生物高分子聚合物,其在骨组织工程材料中的应用已被大量研究报道。虽然海藻酸盐作为骨组织支架材料具有优异的生物相容性、良好的生物可降解性和无免疫原性等优点,但是仍存在机械强度较弱、缺乏细胞特异性结合位点、支架结构在生理环境中易被破坏等缺点,严重限制其在骨组织工程中的应用。目前,研究者已经研究出几大类海藻酸盐复合支架材料,这些材料均表现出优异的力学性能和生化性能,因此,海藻酸盐复合支架材料可能在骨组织修复和再生方面具有较高的应用价值。本文主要针对海藻酸盐复合支架材料在骨组织工程中的应用作简要概述。     

Mutants of avian myelocytomatosis virus with smaller gag gene-related proteins have an altered transforming ability

Avian myelocytomatosis virus strain MC29 is a replication-defective avian oncovirus which in newborn chickens causes myelocytomatosis and liver and kidney tumours. In vitro infection of bone marrow cells gives rise to colonies of transformed macrophage-like cells, and cloned viruses is also capable of transforming fibroblasts. The genome of MC29 contains cellular sequences which are closely related to those in other defective leukaemia viruses with similar transforming spectra. Consequently, these cellular sequences have been postulated to represent a new oncogene which has been designated mac, for macrophage transformation. MC29-transformed cells contain a gag gene-related protein of a 110,000 molecular weight (MW) (p110), which by tryptic peptide analysis has been shown to be a fusion product comprised of a gag gene-derived sequences and sequences which are presumed to be coded by the adjacent mac gene. These findings suggest that this protein may be implicated in transformation by MC29. We now describe three mutants of MC29 and synthesize smaller gag gene-related proteins. These mutants have an altered ability to transform bone marrow cells but not fibroblasts. This demonstrates for the first time a direct involvement of the p110 protein of MC29 in transformation.     

Endothelial and perivascular cells maintain haematopoietic stem cells

Several cell types have been proposed to create niches for haematopoietic stem cells (HSCs). However, the expression patterns of HSC maintenance factors have not been systematically studied and no such factor has been conditionally deleted from any candidate niche cell. Thus, the cellular sources of these factors are undetermined. Stem cell factor (SCF; also known as KITL) is a key niche component that maintains HSCs. Here, using Scf(gfp) knock-in mice, we found that Scf was primarily expressed by perivascular cells throughout the bone marrow. HSC frequency and function were not affected when Scf was conditionally deleted from haematopoietic cells, osteoblasts, nestin-cre- or nestin-creER-expressing cells. However, HSCs were depleted from bone marrow when Scf was deleted from endothelial cells or leptin receptor (Lepr)-expressing perivascular stromal cells. Most HSCs were lost when Scf was deleted from both endothelial and Lepr-expressing perivascular cells. Thus, HSCs reside in a perivascular niche in which multiple cell types express factors that promote HSC maintenance.     

用于软骨下骨修复的镁基支架

骨关节炎（osteoarthritis,OA）作为一种可以导致残疾的退行性疾病,常累及软骨下骨。受损的关节软骨和软骨下骨很难自愈,用于功能修复的组织工程支架是一种有前途的治疗方法。近年来,镁合金因其良好的机械和生物学性能被视为可降解多孔支架有希望的候选者。然而,目前对于适用于软骨下骨缺损修复的镁基支架的结构设计和优化方案还没有定论。归纳了镁合金用于骨软骨支架的研究进展,包括多孔支架的制造方法;添加合金元素和表面改性的优化策略;参数化与非参数化的结构设计;镁基支架的机械、降解和生物学性能及其影响因素。讨论了未来研究的潜在方向。旨在为多孔镁基支架的开发和临床应用提供参考。     

Osteoprotection by semaphorin 3A

The bony skeleton is maintained by local factors that regulate bone-forming osteoblasts and bone-resorbing osteoclasts, in addition to hormonal activity. Osteoprotegerin protects bone by inhibiting osteoclastic bone resorption, but no factor has yet been identified as a local determinant of bone mass that regulates both osteoclasts and osteoblasts. Here we show that semaphorin 3A (Sema3A) exerts an osteoprotective effect by both suppressing osteoclastic bone resorption and increasing osteoblastic bone formation. The binding of Sema3A to neuropilin-1 (Nrp1) inhibited receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation by inhibiting the immunoreceptor tyrosine-based activation motif (ITAM) and RhoA signalling pathways. In addition, Sema3A and Nrp1 binding stimulated osteoblast and inhibited adipocyte differentiation through the canonical Wnt/β-catenin signalling pathway. The osteopenic phenotype in Sema3a?/? mice was recapitulated by mice in which the Sema3A-binding site of Nrp1 had been genetically disrupted. Intravenous Sema3A administration in mice increased bone volume and expedited bone regeneration. Thus, Sema3A is a promising new therapeutic agent in bone and joint diseases.     

Reconstitution with syngeneic plus allogeneic or xenogeneic bone marrow leads to specific acceptance of allografts or xenografts

Clinical organ transplantation between genetically disparate individuals currently requires the use of chemotherapeutic agents to suppress the rejection reaction. The deleterious side effects of these reagents and their inability to prevent rejection completely has led to a continuing search for methods to induce specific transplantation tolerance in adult recipients. Numerous experimental animal models utilizing irradiation and bone marrow transplantation coincident with organ transplantation have been proposed. Bone marrow transplantation, however, has its own major complications, including graft-versus-host reactions and immunoincompetence, probably resulting from a failure of appropriate immune cell interactions in the reconstituted host. We have now attempted to overcome these difficulties by reconstituting the irradiated host with T-cell depleted bone marrow containing both host (syngeneic) and donor (allogeneic or xenogeneic) components. This technique leads to long-term survival of the reconstituted animals and specific prolongation of subsequent skin grafts of donor type. Animals reconstituted in this fashion are fully reactive to third-party allografts and xenografts and do not appear to manifest signs of graft-versus-host disease.     

骨传导听觉脑机接口技术研究

听觉脑机接口技术为视觉通路存在障碍的闭锁综合征患者提供了与外界交流的新通道.为了使空气传导通路受损的病患也能应用此项技术,本文将声音骨传导方式引入到听觉脑机接口实验研究中.本研究设计了左右耳分别呈现不同声音序列的双耳分听实验范式,对骨传导及空气传导方式所诱发脑电信号进行了特征提取和分类比较.实验结果表明两种传导方式下靶刺激均可稳定的诱发出N200与P300电位,骨传导方式下,N200的幅值较空气传导方式更显著.两种方式都获得较高的分类正确率,均可用于听觉脑机接口系统,且骨传导方式优于空气传导方式.     

T-cell specificity for H-2 and Ir gene phenotype correlates with the phenotype of thymic antigen-presenting cells

Experiments with chimaeric animals have demonstrated that the H-2 restriction specificity and immune response (Ir) gene phenotype of the T cell is acquired during development in the thymus. The mechanism by which this process occurs is unclear. One level of obligate expression of H-2 and Ir gene products is on the surface of antigen-presenting cells (APCs) which come from bone marrow precursors. We have now examined the turnover of APCs in the thymuses of F1 leads to parent (P) radiation-induced bone marrow chimaeras and found that APCs of donor phenotype appear at about 2 months after reconstitution. If the peripheral T-cell population is depleted after this time, new T cells emerging from the parental thymus (containing F1 APCs) behaving like F1 T cells, suggesting that cells from the bone marrow can influence thymic-directed T-cell differentiation. The thymic APC is an attractive condidate to play such a part in the development of the T-cell repertoire.     

On Modeling Bio-Scaffolds: Structural and Fluid Transport Characterization Based on 3-D Imaging Data

Bio-scaffolds which are most commonly open celled porous structures are increasingly used for tissue engineering and regenerative medicine. A number of studies have shown that the bulk properties of such irregular structures are poorly modeled using idealized unit cell approaches. The paper therefore uses novel image based meshing techniques to explore both fluid flow and bulk structural properties of a bone scaffold, as accurate modeling of bio-scaffolds with non-uniform cellular structures is very important for the development of optimal scaffolds for tissue engineering application. In this study, a porous hydroxyapatite/tricalcium phosphate (HA/TCP) bone scaffold has been scanned in a Micro-CT scanner, and converted into a volumetric mesh using image processing software developed by the authors. The resulting mesh was then exported to commercial FEA and CFD solvers for analysis. Initial FEA and CFD studies have shown promising results and have highlighted the importance of accurate modeling to understand how microstructures influence the mechanical property of the scaffold, and to analyze flow regimes through the sample. The work highlights the potential use of image based meshing for the ad hoc characterization of scaffolds as well as for assisting in the design of scaffolds with tailored strength, stiffness, and transport properties.     

A Middle Palaeolithic human hyoid bone

The origin of human language, and in particular the question of whether or not Neanderthal man was capable of language/speech, is of major interest to anthropologists but remains an area of great controversy. Despite palaeoneurological evidence to the contrary, many researchers hold to the view that Neanderthals were incapable of language/speech, basing their arguments largely on studies of laryngeal/basicranial morphology. Studies, however, have been hampered by the absence of unambiguous fossil evidence. We now report the discovery of a well-preserved human hyoid bone from Middle Palaeolithic layers of Kebara Cave, Mount Carmel, Israel, dating from about 60,000 years BP. The bone is almost identical in size and shape to the hyoid of present-day populations, suggesting that there has been little or no change in the visceral skeleton (including the hyoid, middle ear ossicles, and inferentially the larynx) during the past 60,000 years of human evolution. We conclude that the morphological basis for human speech capability appears to have been fully developed during the Middle Palaeolithic.     

Epidermal Langerhans cells are derived from cells originating in bone marrow

Langerhans cells constitute a morphologically well characterised subpopulation (3--8%) of mammalian epidermal cells which, in contrast to the bulk of epidermal cells, bear Fc-IgG and C3 receptors, express immune response-associated (Ia) antigens and function as antigen-presenting cells and allogeneic stimulatory cells to primed T lymphocytes. The ontogeny of Langerhans cells has been a subject of considerable debate since their discovery. Although some studies suggest that Langerhans cells are of mesenchymal as opposed to neural or melanocytic origin, direct evidence for this has not been presented. In this study we demonstrate that, after 3 weeks, most of the Langerhans cells (LC) in parenteral skin which had been transplanted on to F1 hybrids were of recipient origin whereas keratinocytes remained of donor origin; this indicates that the LC are derived from a mobile pool of cells. Furthermore, in studies of skin from radiation-induced bone marrow chimaeric animals we found that, depending on the strain combination, up to 80% of the epidermal LC were derived from the bone marrow of the donor animals.     

羊骨胶原纤维的制备与表征

以羊骨为原料，采用盐酸脱除其中的矿物质制备得到胶原纤维。考察了盐酸浓度、体系温度等因素对羊骨胶原纤维制备的影响，并揭示了其影响规律。实验室制备羊骨胶原纤维的最优条件为，在每100 mL酸液中投入25 g羊骨粒的条件下，盐酸溶液浓度0.6 mol/L，体系温度25 ℃，每隔8 h更换酸液。对所制备的胶原纤维进行XRD、FT IR、DSC和SEM测试，结果表明，胶原纤维粗蛋白质量分数为96.54%；体系pH=8.5时，胶原纤维的收缩温度为62.6 ℃；胶原微纤维直径约为50~100 nm，并保持了完整的三股螺旋结构及典型的肽链结构。     

Use-wear analysis confirms the use of Palaeolithic bone tools by the Lingjing Xuchang early human

During 2007–2008 excavations at the Lingjing site near Xuchang, Henan Province, dated back to around 100–80 ka ago, a large quantity of mammalian fossil remains were recovered along with a remarkable cluster of Early Modern Human (EMH) skull fragments in situ. Observably some of those animal bones were probably modified into tools. A use-wear analysis was carried out to examine the functions of modified bone tools. The results suggest that Lingjing bone tools were used for drilling, penetrating, and scraping animal substances, and that some might have been hafted during the use. This study confirms that early existence of intentionally-modified bone tools at human occupations of the early Late Pleistocene in northern China. This discovery suggests making and use of bone tools were inevitably a part of early human behaviors and cultural development, as such of stone tools.     

Fine structure of bone in dinosaurs, birds and mammals

After observation of detailed structural evidence for the origin of birds from dinosaurs, and in light of evidence that dinosaur bone tissue resembles the histology in mammals, the histology of bone has become one of the focal points in discussions of the physiology of dinosaurs and Mesozoic birds. Most of this microstructural information has focused on features related to the vascular organization and the amount of remodelled bone around vascular canals. However, the finer structures have received less attention, although differences in such structures have been observed among modern vertebrates. Here we present evidence that canaliculi--the submicrometre-sized channels that interconnect bone cells and vascular canals--and the collagen fibre bundles in bone are differently organized among certain dinosaur lineages. Ornithomimid dinosaurs are more like birds than mammals in these features. In canalicular structure, and to some extent in fibre bundle arrangement, ornithischian dinosaurs are more like mammals. These differences in both canalicular and lamellar structure are probably linked to differences in the process and rate of bone formation.     

骨髓间充质干细胞向肝样细胞分化的研究进展

摘要: 由各种因素导致的重症肝病的终末治疗的最好手段一直是原位肝移植,但长期以来肝供体的缺乏和免疫排斥引起的一系列问题极大地限制了该手术的运用,同时,在肝脏相关药物的筛选中,原代肝细胞难于培养且易在培养过程中变异,而随着骨髓间充质干细胞研究的深入,越来越多的证据表明骨髓间充质干细胞具有向肝细胞分化的潜能。因此,骨髓间充质干细胞诱导分化而成的肝样细胞在再生医疗和药物筛选领域具有较好的运用前景,本文就间充质干细胞的分离培养及其生物学特性,肝样细胞的诱导培养条件,生物学特性及其运用前景加以综述。     

骨疏康颗粒防治骨质疏松症研究概况

回顾并分析了纯中药制剂骨疏康(GSK)治疗骨质疏松症的基础研究和临床研究相关文献．结果显示：(1)GSK在药理学方面具有特殊性，含有了多种西药的联合效价．(2)在1085例临床报告中(病例选择偏重于绝经期后1型骨质疏松症女性病人)，GSK对绝经期后妇女和1型骨质疏松症病人的疗效可靠性高于其他类型(包括男性、合并压缩骨折，高龄等)；通过骨密度测定，GSK可以明显增加骨密度(BMD)，即提高骨量，但对于改善骨的体积密度(BVD)，即改善骨质的效应尚不能明确肯定；GSK临床疗效和安全性稳定可靠，所有文献均有较满意的正向结论，可大致归结为：3个月疗程，75％的显效率．     

从骨胶水解液中分离L-羟脯氨酸和L-脯氨酸

以骨胶为原料，进行酸水解，用亚硝酸钠氧化，７３２阳离子交换树脂层析分离提取Ｌ－羟脯氨酸和Ｌ－脯氨酸。得到纯的Ｌ－羟脯氨酸和Ｌ－脯氨酸均在（７．１～７．２）％左右，产品质量均达到出口标准。