Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells

Rat calvaria osteoblasts derived from 21-day-old fetal rat pups undergo a temporal expression of markers of the osteoblast phenotype during a 5 week culture period. Alkaline phosphatase and osteocalcin are sequentially expressed in relation to collagen accumulation and mineralization. This pattern of expression of these osteoblast parameters in cultured rat osteoblasts (ROB) is analogous to that seen in vivo in developing fetal rat calvaria tissue (Yoon et. al: Biochem. Biophis. Res. Commun. 148:1129, 1987) and is similar to that observed in cultures of subcultivated 16-day-old embryonic chick calvaria-derived osteoblasts (COB) (Gerstenfeld, et.al: Dev. Biol. 122:46, 1987). While the cellular organization of subcultivated COB and primary ROB cultures are somewhat different, the temporal expression of the parameters remains. Both the rat and chick culture systems support formation of matrix mineralization even in the absence of beta-glycerol-phosphate. A systematic examination of factors which constitute conditions supporting complete expression of the osteoblast phenotype in ROB cultures indicate requirements for specific serum lots, ascorbic acid and the ordered deposition of mineral in the extracellular matrix. The present studies suggest that formation of a collagenous matrix, dependent on ascorbic acid, is requisite for expression of the osteoblast phenotype. In ROB cultures, expression of osteocalcin synthesis occurs subsequent to initiation of alkaline phosphatase activity and accompanies the formation of mineralized nodules. Thus, extracellular matrix mineralization (deposition of hydroxyapatite) is required for complete development of the osteoblast phenotype, as reflected by a 200-fold increase in osteocalcin synthesis. These data show the temporal expression of the various osteoblast parameters during the formation and mineralization of an extracellular matrix can provide markers reflective of various stages of osteoblast differentiation/maturation in vitro.     

大熊猫生殖生物学研究:Ⅱ.大熊猫妊娠期尿中孕酮和绒毛膜促性腺激素样物质含量的变化

本文用放射免疫法测定了四只大熊猫在妊娠期和非妊娠期尿中孕酮和CG样物质含量的变化。结果表明:在大熊猫胚胎着床前,孕酮和CG样物质含量较低;胚胎着床后,二种激素的含量显著上升并达到高峰。雌兽妊娠期尿中孕酮和CG样物质含量的变化规律可为证实大熊猫系延缓着床型动物提供一些理论依据;同时,也为大熊猫的妊娠诊断和预测分娩提供一些参考数据。     

反义寡核苷酸片段可抑制鲫鱼视网膜电压依赖性钾离子通道的表达

爪蟾卵母细胞经注射鲫鱼视网总ＲＮＡ后,可表达大量电压依赖性钾离子通道。在此基础上,我们进一步发现,一个特定序列的寡核苷酸能专一地抑制该通道的表达。由于我们设计与合成的该片段与果蝇、小鼠中已克隆的钾通道中编码Ｎ端的一个多肽的ｍＲＮＡ完全互补,因此推测：鲫鱼视网膜中的Ｋ这个区域与其它物种的Ｋ通道有高度同源性,这为克隆该基因和研究它的功能提供了重要资料。     

蓝藻藻胆体与类囊体膜之间光能传递的研究

从嗜热蓝藻优雅粘囊藻（Ｍｙｘｏｓａｒｃｉｎａ ｃｏｎｃｉｎｎａ Ｐｒｉｎｔｚ．）中分离到具有放氧活性的藻胆体－类囊体膜复合物。它的吸收峰位于６８０、６２８、４９０、４３８和４２０ ｎｍ ,在低离子强度（０．１～０．４ ｍ ｏｌ／Ｌ）磷酸缓冲液中的６６４ ｎｍ 荧光发射峰随离子强度的降低而升高,７１８ ｎｍ 荧光发射峰与此相反（７７ 。Ｋ,Ｅｘ＝ ５８０ ｎｍ ）。当把游离的藻胆体和已解离去藻胆体的类囊体膜在蔗糖磷酸缓冲液中重组时,随时间延长（０～６０ ｍ ｉｎ）,７１８ ｎｍ 荧光发射峰逐渐升高,６８５ ｎｍ 荧光发射峰逐渐下降（７７ 。Ｋ,Ｅｘ ＝ ５８０ ｎｍ ）,表明藻胆体与类囊体之间的解离和结合对光能传递的影响     

力竭游泳对大鼠心肌线粒体钙运输的影响

 力竭游泳对大鼠心肌线粒体钙运输的影响丁树哲,王文信,连克杰,许豪文（华东师范大学体育系运动生化实验室,上海２０００６２）线粒体钙运输在细胞功能调节方面有重要作用．线粒体通过摄取与释放钙,对其跨膜质子、不依赖底物及产物抑制的ＡＴＰ合成、磷酸化偶联等均有...     

TGFβ alters growth and differentiation related gene expression in proliferating osteoblasts in vitro,preventing development of the mature bone phenotype

This study examines the mechanism by which TGF-β1, an important mediator of cell growth and differentiation, blocks the differentiation of normal rat diploid fetal osteoblasts in vitro. We have established that the inability for pre-osteoblasts to differentiate is associated with changes in the expression of cell growth, matrix forming, and bone related genes. These include histone, jun B, c-fos, collagen, fibronectin, osteocalcin, alkaline phosphatase, and osteopontin. Morphologically, the TGF-β1-treated osteoblasts exhibit an elongated, spread shape as opposed to the characteristic cuboidal appearance during the early stages of growth. This is followed by a decrease in the number of bone nodules formed and the amount of calcium deposition. These effects on differentiation can occur without dramatic changes in cell growth if TGF-β1 is given for a short time early in the proliferative phase. However, continuous exposure to TGF-β1 leads to a bifunctional growth response from a negative effect during the proliferative phase to a positive growth effect during the later matrix maturation and mineralization phases of the osteoblast developmental sequence. Extracellular matrix genes, fibronectin, osteopontin and α1(I) collagen, are altered in their expression pattern which may provide an aberrant matrix environment for mineralization and osteoblast maturation and potentiate the TGF-β1 response throughout the course of osteoblast differentiation. The initiation of a TGF-β1 effect on cell growth and differentiation is restricted to the proliferative phase of the culture before the cells express the mature osteoblastic phenotype. Second passage cells that are accelerated to differentiate by the addition of dexamethasone or by seeding cultures at a high density are refractory to TGF-β1. These in vitro results indicate that TGF-β1 exerts irreversible effects at a specific stage of osteoblast phenotype development resulting in a potent inhibition of osteoblast differentiation at concentrations from 0.1 ng/ml. © 1994 Wiley-Liss, Inc.     

Differential effects of warfarin on mRNA levels of developmentally regulated vitamin K dependent proteins,osteocalcin, and matrix Gla protein in vitro

The role of the vitamin K dependent proteins, osteocalcin which is bone specific and matrix Gla protein (MGP) found in many tissues, has been studied by inhibition of synthesis of their characteristic amino acid, γ-carboxyglutamic acid (Gla) with the anticoagulant sodium warfarin. The effect of sodium warfarin on expression of these proteins, and other phenotypic markers of bone and cartilage during cellular differentiation and development of tissue extracellular matrix, was examined in several model systems. Parameters assayed include cell growth (reflected by histone gene expression) and collagen types I and II, osteopontin, alkaline phosphatase, and mineralization. Studies were carried out in calvarial bone organ cultures, normal diploid rat osteoblast and chondrocyte cultures, and rat osteosarcoma cell lines ROS 17/2.8 and 25/1. In normal diploid cells, warfarin consistently stimulated cell proliferation (twofold). In osteoblast cultures, MGP mRNA levels were generally increased (three to tenfold). Notably, MGP mRNA levels were not affected in chondrocyte cultures, either with chronic or acute warfarin treatments. Osteocalcin mRNA levels and synthesis were decreased up to 50% in ROS 17/2.8 cells and in chronically treated (1 and 5 μg/ml sodium warfarin) rat osteoblast cultures after 22 days. Early stages of osteoblast phenotype development from the proliferation period to initial tissue formation (nodules) appeared unaffected; while after day 14, further growth and mineralization of the nodule areas were significantly decreased in warfarin-treated cultures. In summary, warfarin has opposing effects on the expression of two vitamin K dependent proteins, MGP and osteocalcin, in osteoblast cultures and MGP is regulated differently between cartilage and bone as reflected by cellular mRNA levels. Additionally, warfarin effects expression of nonvitamin K dependent proteins which may reflect the influence of warfarin on endoplasmic reticulum associated enzymes. © 1994 Wiley-Liss, Inc.     

柱萤叶甲属研究（Ⅰ）鞘翅具黑色刻点的种类记述（鞘翅目：叶甲科）

本文就萤叶甲亚科中柱萤叶甲属鞘翅具黑色刻点的种类进行研究,共记述４种,我国已记录３种,其中１种为新种。