Effects of lanthanum and gadolinium on proliferation and differentiation of primary osteoblasts

The effects of La³⁺ and Gd³⁺ on the proliferation, differentiation and adipogenic transdifferentiation of rat calvarial osteoblast-like cells (ROB cells) were evaluated by MTT method， measuring the activity of alkaline phosphatase (ALP) and Oil red O measurement. Both of La³⁺ and Gd³⁺ inhibited the proliferation of ROB cells at all concentrations (1×10^-5, 1×10^-6, 1×10^-7, 1×10^-8, 1×10^-9 mol?L^-1). La³⁺ at concentration of 1×10^-5 mol?L^-1 significantly increased the alkaline phosphatase activity of ROB cells up to 3 folds (P<0.01). However, the effects reversed to inhibit at other concentrations. Gd³⁺ played a negative role in the alkaline phosphatase activity. La³⁺ inhibited the adipogenic transdifferentiation of ROB cells at all concentrations in a dose-dependent way. However, Gd³⁺ promoted the adipogenic transdifferentiation of ROB cells at 1×10^-8 and 1×10^-9 mol?L^-1. These findings suggested that the effects of rare earth elements on the proliferation, differentiation and adipogenic transdifferentiation of ROB cells were dependent on their concentrations and species     

Effects of Nd<Superscript>3+</Superscript> and Sm<Superscript>3+</Superscript> on the proliferation,differentiation and mineralization function of primary osteoblasts <Emphasis Type="Italic">in vitro</Emphasis>

A series of experimental methods including 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) test, alkaline phosphatase (ALP) activity measurement, Oil Red O stain and measurement, mineralized function expression and quantitive real time RT-PCR (qRT-PCR) were employed to assess the effect of Nd³⁺ and Sm³⁺ on the proliferation, differentiation and mineralization function of primary osteoblasts (OBs) in vitro at cell and molecular levels. The experimental results suggest that concentration, culture time and ion species are pivotal factors for switching the biological effects of rare earth ions from toxicity to activity, from damage to protection, or from down-regulation to up-regulation.     

去乙酰化酶抑制剂TSA对间充质干细胞C3H10T1/2增殖和成脂分化的影响

采用MTT和流式细胞术分别检测不同浓度的TSA对C3H10T1/2细胞活性和细胞周期分布的影响;油红O染色检测TSA对其成脂分化的影响,实时定量PCR检测TSA对成脂分化的关键转录因子PPAR-γ,以及成脂分化标志物Fabp4和Adipoq mRNA转录的影响.研究去乙酰化酶抑制剂TSA对间充质干细胞C3H10T1/2增殖和成脂分化的影响及其可能的作用机制.结果显示TSA浓度为1、10和30 nmol/L呈浓度依赖性地抑制C3H10T1/2细胞活性,改变细胞形态,并将其细胞周期抑制在G0/G1期;TSA浓度为10nmol/L明显抑制C3H10T1/2细胞的成脂分化作用,并呈浓度依赖性地抑制PPAR-γ、Fabp4和Adipoq mRNA的转录.表明TSA呈剂量依赖性地抑制间充质干细胞C3H10T1/2的增殖和成脂分化,除转录水平调控外,非组蛋白如细胞骨架相关蛋白可能也参与TSA的抑制作用.     

生物力学作用对骨髓间充质干细胞增殖和成骨分化影响研究进展

骨髓间充质干细胞在体内所处的力学环境比较复杂,为了探索生物力学与骨髓间充质干细胞增殖分化之间的关系,对近年来生物力学微环境对骨髓间充质干细胞增殖分化影响的研究现状和最新进展进行了综述。认为牵张力与流体剪切力对骨髓间充质干细胞产生的刺激大部分会使其向成骨方向分化,而较大的压缩力和静水压力对骨髓间充质干细胞产生的刺激大部分会使其偏向软骨方向分化,小部分向成骨方向分化,每种分化方向都有其最适的分化条件。通过综述生物力学对骨髓间充质干细胞增殖及成骨分化的影响,为骨髓间充质干细胞的骨组织工程与再生医学研究及更好地应用于临床治疗提供思路和参考依据。     

马钱子苷对神经干细胞的增殖、存活和分化的调节作用

为探讨不同浓度的马钱子苷对神经干细胞的增殖、存活和分化的调节作用及其相关分子机制,本实验从成年小鼠大脑中分离培养了神经干细胞,用不同浓度的马钱子苷进行干预,观察马钱子苷对神经干细胞增殖、存活和分化的影响。结果显示：成年小鼠神经干细胞在含有中、高浓度马钱子苷的增殖培养基中培养5 d和7 d后,神经球数量和直径与对照相比显著增加（P<0.05）;中、高剂量的马钱子苷能够促进神经干细胞的有丝分裂,低剂量马钱子苷处理显著促进神经干细胞发生分化（P<0.01）,并增加神经元和星形胶质细胞的数量及比例（P<0.01）;中、高剂量马钱子苷抑制神经干细胞分化（P<0.05）,高剂量的马钱子苷使得神经元的数量减少（P<0.05）。研究结果表明,高浓度的马钱子苷能够促进神经干细胞存活,并通过促进神经干细胞有丝分裂来提高其增殖能力;低浓度的马钱子苷促进神经干细胞分化,有利于神经再生和少突胶质细胞再生。研究结果为神经干细胞治疗中枢神经系统疾病的研究奠定了理论和实验基础。     

Effect of Dy3+on osteogenic and adipogenic differentiation of mouse primary bone marrow stromal cells and adipocytic trans-differentiation of mouse primary osteoblasts

A series of experimental methods including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test, alkaline phosphatase (ALP) activity measurement, mineralized function, Oil Red O stain and measurement were employed to assess the effect of Dy³⁺ on the osteogenic and adipogenic differentiation of mouse primary bone marrow stromal cells (BMSCs) and the adipogenic trans-differentiation of mouse primary osteoblasts (OBs). The results showed that Dy³⁺ had no effect on BMSC proliferation at concentrations of 1×10⁻⁸ and 1×10⁻⁵ mol/L, but inhibited BMSC proliferation at other concentrations. Dy³⁺ had no effect on OB proliferation at concentrations of 1×10⁻¹⁰ and 1×10⁻⁹ mol/L, but inhibited OB proliferation at other concentrations. Dy³⁺ had no effect on the osteogenic differentiation of BMSCs at concentrations of 1×10⁻⁹ and 1×10⁻⁷ mol/L, and promoted osteogenic differentiation of BMSCs at other concentrations at the 7th day. The osteogenic differentiation of BMSCs was inhibited by Dy³⁺ at concentration of 1×10⁻⁵ mol/L at the 14th day, but promoted osteogenic differentiation of BMSCs at concentrations of 1×10⁻⁹, 1×10⁻⁸, 1×10⁻⁷ and 1×10⁻⁶ mol/L with the maximal effect at concentration of 10⁻⁶ mol/L. Dy³⁺ promoted mineralized function of BMSCs at any concentration. Dy³⁺ had no effect on adipogenic differentiation of BMSCs at concentration of 1×10⁻⁷ mol/L, but inhibited adipogenic differentiation of BMSCs at other concentrations. Dy³⁺ inhibited adipocytic trans-differentiation of OBs at any concentration, suggesting that Dy³⁺ had protective effect on bone and the protective effect on bone may be mediated by modulating differentiation of BMSCs away from the adipocyte and inhibiting adipocytic trans-differentiation of OBs which may promote differentiation and mineralization of OBs. These results may be valuable for better understanding the mechanism of the effect of Dy³⁺ on pathogenesis of osteoporosis. Supported by the Foundation for Key Program of Ministry of Education of China (Grant No. 208018)     

Effects of low frequency electromagnetic fields on osteoblasts proliferation and cell cycle

After primary mouse osteoblasts and ROS osteo-like cells were exposed to 50 Hz low frequency electromagnetic fields (EMF), the MTT method and flow cytometry have been used to determine cell proliferation, cell cycle and apoptosis. The results show that, compared with the control, the cells after exposure are more abundant, have larger S phase ratios and lower apoptosis ratios. The results indicate that EMF has an important biological effect which influences cell proliferation and cell cycle.     

Effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of bone marrow stromal cells and on the adipogenic trans-differentiation of osteoblasts

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), alkaline phosphatase (ALP) activity and oil red O assays were used to examine the effects of genistein, daidzein and glycitein on the osteogenic and adipogenic differentiation of primary mouse bone marrow stromal cells (MSCs) and the adipogenic trans-differentiation of primary mouse osteoblasts. The results indicated that daidzein, genistein and glycitein at concentrations from 1×10^-8 mol/L to 1×10^-5 mol/L promoted the proliferation of MSCs and osteoblasts; genistein, daidzein and glycitein promoted osteogenic differentiation and inhibited adipogenic differentiation of MSCs, and inhibited adipocytic transdifferentiation of osteoblasts at appropriate concentrations as 17β-estradiol. It suggests that genistein, daidzein and glycitein regulate a dual differentiational process of MSCs into the osteogenic and adipogenic lineages, and trans-differentiational process of primary osteoblasts into the adipocyte lineages, causing a lineage shift toward osteoblast. Protective effects of them on bone may be mediated by a reversal of adipogenesis which may promote the proliferation, differentiation and mineralization of osteoblasts, and make adipocytes secrete less cytokines which may promote osteoclast formation and activation. In addition, the results also indicated that genistein, daidzein and glycitein may be helpful in preventing the development of steroid induced osteonecrosis.     

阿维A对HaCaT细胞增殖的影响

目的:探讨阿维A对角质形成细胞(HaCaT)增殖的影响及其发挥作用的适宜浓度.方法:分别以浓度为1×10-7mol/L、1×10-6mol/L、5×10-6mol/L阿维A作用于HaCaT细胞,用四氮唑盐类化合物(MTS)比色法检测每组12、24、48和72 h HaCaT细胞的增殖情况.结果:与空白对照组相比,大于或等于1×10-6mol/L阿维A可以抑制HaCaT细胞增殖,并且在作用24 h后随着阿维A浓度的增加,其对HaCaT细胞增殖的抑制作用增强.结论:阿维A抑制HaCaT细胞增殖适宜浓度为5×10-6mol/L.     

Cytoskeleton reorganization and FAK phosphorylation are involved in lanthanum(Ⅲ)-promoted proliferation and differentiation in rat osteoblasts

The effect of lanthanum ion(La3+)on osteoblast function and cytoskeleton is assessed in vitro. Osteoblasts were isolated from Snraeuo-tjawiey tetai neonatal rats.Cell proliferation and gene expression levels of cbfa-1,alkaline phosphatase(ALP),osteocalcin(OC),bone sialo-protein(BSP)and osteopontin(OPN)were examined by cell counting and RT-PCR.Cytoskeleton F-actin was stained with rhodamine-con-jugated phalloidin and was visualized by a confocal microscope.As the results,10-8-10-4M La3+-induced osteoblast proliferation on day2.Data from the RT-PCR assay revealed that 10-6 M La3+ up-regulated the expression levels of ALP,BSP,and cbfa-lon day 4,while it enhanced the expressions of OC and OPN on day 21.The F-actin cytoskeleton was strengthened and reorganized under the exposure of La3+.In addition,the phosphorylation of focal adhesion kinase(FAK)was significantly promoted in 24 h evaluated by Western blot analysis.These findings indi-sate that La 3+ promotes osteoblast activity through the phosphorylation of F AK and reorganization of the cytoskeleton.     

Cytoskeleton reorganization and FAK phosphorylation are involved in lanthanum （Ⅲ）-promoted proliferation and differentiation in rat osteoblasts

The effect of lanthanum ion (La³⁺) on osteoblast function and cytoskeleton is assessed in vitro. Osteoblasts were isolated from Sprague-Dawley fetal neonatal rats. Cell proliferation and gene expression levels of cbfa-1, alkaline phosphatase (ALP), osteocalcin (OC), bone sialoprotein (BSP) and osteopontin (OPN) were examined by cell counting and RT-PCR. Cytoskeleton F-actin was stained with rhodamine-conjugated phalloidin and was visualized by a confocal microscope. As the results, 10⁻⁸-10⁻⁴ M La³⁺-induced osteoblast proliferation on day 2. Data from the RT-PCR assay revealed that 10⁻⁶ M La³⁺ up-regulated the expression levels of ALP, BSP, and cbfa-1 on day 4, while it enhanced the expressions of OC and OPN on day 21. The F-actin cytoskeleton was strengthened and reorganized under the exposure of La³⁺. In addition, the phosphorylation of focal adhesion kinase (FAK) was significantly promoted in 24 h evaluated by Western blot analysis. These findings indicate that La³⁺ promotes osteoblast activity through the phosphorylation of FAK and reorganization of the cytoskeleton.     

人骨髓间充质干细胞不同亚群

目的 比较不同亚群的人骨髓间充质干细胞(human bone mesenchymal stem cell, hBMSC)自我更新和分化能力。方法 通过获赠的人髂骨骨髓标本,联合运用密度梯度离心和差异贴壁法分离MSCs , 用10μm 滤膜将不同群体细胞分离,倒置相差显微镜观察不同亚群细胞的形态;流式细胞仪检测BMSC不同亚群细胞的表型;在地塞米松、Vit C、β-磷酸甘油钠作用下将不同亚型细胞向成骨细胞诱导分化,分别观察其分化能力。结果 成熟的MSC,即mMSCs 细胞(mature cells)呈纤维样梭形, RS 细胞(rapidly MSC self-renewing cells)呈圆形。RS细胞增殖能力明显强于mMSCs。经定向诱导分化后,RS细胞向成骨细胞分化能力较mMSCs细胞强。结论 RS 细胞较之mMSC细胞可能是一种更原始的中胚层前体细胞,具有更强的自我更新和分化潜能。     

硝酸镧对流苏石斛试管苗生长的影响

探讨了不同浓度的硝酸镧对流苏石斛试管苗生长的影响。结果表明,硝酸镧对流苏石斛试管苗的增殖率、高生长、茎粗、叶长、叶宽、生根数、生根率、根长、质量增量、硝酸还原酶活性、过氧化物酶活性等有显著影响。硝酸镧质量浓度低于10 mg/L时能有效促进流苏石斛试管苗的生长分化。     

胆汁酸及FXR在细胞增生和分化中的作用研究进展

 胆汁酸具有多种重要的生理功能,不仅与脂类物质的消化吸收密切相关,还可作为激素样信号分子在胆汁酸、脂类与糖类物质代谢及能量代谢等方面发挥重要作用。近年研究发现,胆汁酸及其核受体法尼酯X受体（FXR）在细胞增生、分化、凋亡和肝再生的调控中也发挥重要作用。本文综述了胆汁酸及其核受体FXR在细胞增生、分化、凋亡和肝再生的作用及机制,以及在寄生虫-细粒棘球绦虫成虫发育分化中作用的研究进展。     

ROZ与ATRA联合应用对MCF-7细胞诱导作用

研究PPART激动剂罗格列酮（ROZ）与全反式维甲酸（ATRA）联合应用诱导MCF-7细胞发生分化和凋亡的作用,阐述PPARγ激活后的抗肿瘤作用和机制．采用克隆原形成实验测定两化合物对MCF-7细胞存活率的影响;采用诱导分化实验测定两化合物对MCF-7细胞分化的影响;并进一步应用RT—PCR分析两化合物对MCF-7细胞相关基因表达的影响．集落形成实验显示,两化合物联合应用后,能明显抑制MCF-7细胞的生长．诱导分化实验表明,ROZ在低剂量时促进MCF-7细胞的分化,而随着剂量的增加,分化作用减弱,与ATRA联合应用后,同样在低剂量时促进MCF-7细胞分化,并且出现脂滴的细胞数增加,脂滴增大,在高剂量时,分化作用亦有所减弱。RT—PCR结果表明,在两化合物的作用下,MCF-7细胞中PPARγ的表达升高,并使MCF-7细胞中与凋亡相关基因的表达发生变化．ROZ与ATRA联合应用具有协同作用,激活PPARγ,抑制MCF-7细胞增殖,诱导MCF-7细胞发生分化．     

Effects of gadolinium addition on the microstructure and mechanical properties of Mg-9Al alloy

This research aims to study the significance of Gd addition (0wt%-2wt%) on the microstructure and mechanical properties of Mg-9Al alloy. The effect of Gd addition on the microstructure was investigated via X-ray diffraction (XRD), optical microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The Mg-9Al alloy contained two phases, α-Mg and β-Mg17Al12. Alloying with Gd led to the emergence of a new rectangular-shaped phase, Al₂Gd. The grain size also decreased marginally upon Gd addition. The ultimate tensile strength and microhardness of Mg-9Al alloy increased by 23% and 19%, respectively, upon 1.5wt% Gd addition. We observed that, although Mg-9Al-2.0Gd alloy exhibited the smallest grain size (181 μm) and the highest dislocation density (5.1×1010 m-2) among the investigated compositions, the Mg-9Al-1.5Gd alloy displayed the best mechanical properties. This anomalous behavior was observed because the Al₂Gd phase was uniformly distributed and present in abundance in Mg-9Al-1.5Gd alloy, whereas it was coarsened and asymmetrically conglomerated in Mg-9Al-2.0Gd.     

驱蚊香草愈伤组织的诱导和增殖

以驱蚊香草（Pelargonium graveolens）的幼嫩叶片为外植体,接种于附加不同浓度的NAA、2,4-D、6-BA、KT及其组合的MS固体培养基上,结果表明：单独使用4种植物生长调节物质对驱蚊香草愈伤组织的诱导在一定浓度范围内都有效果,其中单独使用KT时效果不太明显;最佳的组合培养基为MS＋2,4-D0．2mg／L＋6BA0．5mg／L．在黑暗和光照培养条件下．愈伤组织的增殖呈“S”型,且生长周期均为30d．     

Distribution of lanthanum among the chloroplast subcomponents of spinach and its biological effects on photosynthesis： location of the lanthanum binding sites in photosystem Ⅱ

The effects of lanthanum at different concentrations on the related photosynthetic activities of Hill reaction, Mg^2＋-ATPase and Ca^2＋-ATPase in spinach chloroplast were studied. Experimental results showed that lanthanum can increase all the activities at suitable concentration （15-30 mg· L^-1）, however, it behaves toxically on them when over used （60 mg. L^-1）. To get an improved understanding of the mechanism of lanthanum effects on the photosynthesis of spinach, the different subcomponents in the chloroplast of the cultured spinach were isolated, and the content of lanthanum in each subcomponent was determined by ICP-MS. The results obtained indicated that among these different subcomponents, about 90% out of the total chloroplast lanthanum was located in photosystem Ⅱ （PS Ⅱ） while there was little lanthanum in photosystem Ⅰ （PS Ⅰ）. Moreover, size exclusion high performance liquid chromatography （SE-HPLC） coupled with online UV and ICP-MS detections was novelly used for locating lanthanum binding sites in PS Ⅱ proteins for the first time. It was found that lanthanum has two binding sites in PS Ⅱ： La associates with chlorophyll together with magnesium in PS Ⅱ by partly replacing magnesium and also shares the common binding sites of PS Ⅱ proteins together with the inorganic cofactors of calcium and manganese, influencing the process of photosynthesis.