Marrow mesenchymal stem cells transduced with TPO/FL genes as support for ex vivo expansion of hematopoietic stem/progenitor cells

A new marrow-derived mesenchymal stem cell (hMSC) line that could support expansion of hematopoietic stem/progenitor cells (HSPCs) was developed. Primary hMSCs were infected with retrovirus containing Flt-3 ligand and thrombopoietin genes. CD34+ cells from cord blood were expanded with primary hMSCs or transduced hMSCs. The expansion of total nucleated cells, CD34+ cells and mixed colonies containing erythroid and myeloid cells and megakaryocytes for 2 weeks coculture with transduced hMSCs was remarkably increased. The outputs of long-term culture-initiating cells for 2 and 4 weeks coculture with transduced hMSCs were also largely increased. The expansion rates of HSPCs with transduced hMSCs were unchanged for 6 weeks. In contrast, the expansion rates of HSPCs with primary hMSCs declined drastically through 6 weeks. SCID-repopulating cell expansion with transduced hMSCs for 4 weeks was significantly higher than that of uncultured CD34+ cells and HSPCs expanded with primary hMSCs. Received 21 June 2005; received after revision 30 July 2005; accepted 24 August 2005     

Elucidation of the relationship between enzyme activity and internal motion using a lysozyme stabilized by cavity-filling mutations

We investigated the activity and the internal motions of a stabilized mutant hen lysozyme (HEL) in which the residues M12 and L56 were mutated to L and F, respectively (LF mutant HEL). The result of the activity measurements against glycol chitin at various temperatures suggested that the temperature dependence of the activity of LF mutant HEL shifted to the high-temperature side compared with that of wild-type HEL. The detailed internal motions of LF mutant HEL in the absence and presence of a substrate analogue, (NAG)₃, were examined by model-free analysis at 35°C. The results showed that the internal motions of LF mutant HEL in the presence of (NAG)₃ were drastically restricted compared with those in wild-type HEL. Our findings thus suggested that the mutation to the stabilized lysozyme restricted internal motions required for the enzymatic reaction.Received 8 February 2005; accepted 10 March 2005Y. Yoshida and T. Ohkuri contributed equally to this work.     

Lactoferrin

Mammalian lactoferrin (Lf) receptors are suggested to have pivotal roles for mediating multiple functions of Lf. In this review, we focus on current knowledge of the structure and function of mammalian Lf receptors, mainly the first cloned Lf receptor that has been shown to be expressed in the infant small intestine at high levels but also in virtually all other tissues. The small intestinal Lf receptor takes up iron from Lf into cells and presumably exerts other physiological functions. Other Lf receptors in various tissues have also been reported to mediate some functions of Lf, such as modulating immune function, inhibiting platelet aggregation and enhancing collagen gel contractile strength. The detailed mechanisms behind the receptor-Lf interactions still need to be elucidated.     

Compensatory caspase activation in MPP+-induced cell death in dopaminergic neurons

Many have hypothesized that cell death in Parkinsons disease is via apoptosis and, specifically, by the mitochondrial-mediated apoptotic pathway. We tested this hypothesis using a mouse dopaminergic cell line of mesencephalic origin, MN9D, challenged with the Parkinsonism-causing neurotoxin MPP⁺ (1-methyl-4-phenylpyridinium ion). Apoptosis was the main mode of cell death when the cells were subjected to MPP⁺ treatment under serum-free conditions for 24 h. Caspase-3 and caspase-9, however, were not activated, thus indicating the existence of alternate or compensatory cell death pathway(s) in dopaminergic neuronal cells. Using caspase inhibitors, we demonstrated that these pathways involve caspase-2, –8, –6 and –7. A time-course study indicated that activation of caspase-2 and –8 occurred upstream of caspase-6 and caspase-7. Upon MPP⁺ challenge, the apoptosis-inducing factor was translocated from the mitochondria into the MN9D cytosol and nucleus. These results suggest the existence of alternative apoptotic pathways in dopaminergic neurons.Received 20 September 2004; received after revision 5 November 2004; accepted 22 November 2004     

姜酚类化合物的稳定性及其抗氧化性能研究3

以生姜为原料,采用超临界CO2萃取姜酚类化合物.在强酸性条件下姜酚类化合物脱水成姜烯酚后,与甘氨酸加成生成姜烯酚一甘氨酸复合物以增强姜酚类化合物的稳定性.用紫外分光光度法和差分脉冲伏安法对姜烯酚-甘氨酸进行研究.结果表明:紫外分光光度法测定姜烯酚-甘氨酸,最大特征吸收峰在230、280 nm处.吸光度比值为1.776;差分脉冲伏安法测定姜烯酚-甘氨酸,以玻碳电极为工作电极,在pH=2～8的范围内,姜烯酚-甘氨酸的电化学活性随着pH的降低而减弱;姜烯酚-甘氨酸对N,N-二苯基N'-苦味基肼基自由基(DPPH·自由基)具有良好的清除作用,其对DPPH·自由基的清除率随计量的增加而增加,当加入量为0.125 mg时,其清除率为81.8%.研究发现,姜烯酚-甘氨酸对DPPH·自由基的清除能力高于甘氨酸.     

同伦近似对称法：六阶Boussinesq方程的同伦级数解

提出了用以处理非线性问题的同伦近似对称法,并利用该方法研究流体动力学中的六阶Boussinesq方程.各阶相似约化解和各阶相似约化方程均可以写出通式,从而导出相应的同伦级数解.零阶相似约化方程等价于Painlevé IV型方程或Weierstrass椭圆方程,高阶相似解可以通过解线性变系数常微分方程得到.辅助参数具有调节同伦级数解的收敛性的作用.由近似对称法得到的级数解和各阶相似约化方程均能够由同伦近似对称法重新得到.     

腾冲嗜酸两性菌硫氧化还原酶分子结构中铁原子结合位点的新特征

前期研究表明, 铁原子对于嗜酸两性菌中硫氧化还原酶(SOR)的活性至关重要. 本研究表明, 2,2′-联吡啶、1,2-二羟基苯-3,5-二磺酸钠、8-羟基喹啉等特异性铁离子螯合剂强烈抑制腾冲嗜酸两性菌SOR酶活性, 进一步表明铁原子是SOR酶活必需. 对目前基因组数据库中的SOR基因或者SOR类似基因进行序列比对, 发现了一个潜在的铁原子结合模体(H⁸⁶-X₃- H⁹⁰-X_n-E¹¹⁴-X_n-E¹²⁹). 据此, 本研究采用定点突变技术, 将氨基酸残基H⁸⁶, H⁹⁰和E¹²⁹分别突变为苯丙氨酸或者丙氨酸, 圆二色光谱测定发现突变体(H⁸⁶F, H⁹⁰F和E¹²⁹A)的二级结构没有明显改变, 但是这3个突变体全部丧失了酶活性. 突变体蛋白中铁原子含量测定结果表明, 3个突变体全部或者部分丢失了铁原子, 而之前研究中获得的3个半胱氨酸突变体(完全丧失了酶活)铁原子含量没有变化. 根据本研究并结合前期实验结果可知, SOR分子中模体C³¹-X_n-C¹⁰¹-X₂-C¹⁰⁴是底物硫分子活化区域; 而模体H⁸⁶-X₃-H⁹⁰-X₂₃-E¹¹⁴-X_¹⁴-(E/D)¹²⁹是SOR分子中铁原子的结合区域, 与铁原子结合形成一个非卟啉铁中心, 是SOR的氧化还原中心; 这两个区域均是SOR酶活性的必需部分.     

乳腺癌血清多肽图研究

乳腺癌一直是全世界范围内威胁妇女健康的恶性疾病, 尽管人们已经进行了大量的研究以减少乳腺癌对人类的危害, 但是乳腺癌仍然是目前导致死亡的恶性肿瘤之一. 乳腺癌的早期发现对于患者的愈后与生存意义重大, 可以明显提高病人生存时间、降低病人的死亡率. 据统计, 在过去的5年里, 早期诊断每年可减少3.2%因乳腺癌死亡的患者. 然而研究表明, 目前常用的乳腺癌诊断技术, 如乳腺X射线摄影和乳房检查均无法诊断出40%的早期乳癌患者和大多数年轻女性的乳腺肿瘤. 因此在乳腺癌临床治疗中, 急需发展新型的高效诊断技术.     

GaInP/GaAs/Ge三结叠层光电池光谱响应的温度特性

利用金属有机物化学气相沉积法(MOCVD)制备了转化效率达27.1%的GaInP/GaAs/Ge三结叠层电池, 并对其光谱响应的温度特性进行了测量研究. 通过光谱响应曲线观察到各子电池的吸收边随温度升高发生红移, 这主要归因于电池材料禁带宽度的变窄效应. 根据光谱响应数据计算得到的GaInP/GaAs/Ge叠层电池各子电池在室温下的短路电流密度分别为12.9, 13.7和17 mA/cm², 且叠层电池的短路电流密度的温度系数为8.9 μA/(cm²·℃). 最后, 根据叠层电池的串联结构推导了其电压温度系数为-6.27 mV/℃.     

冷热交替治疗对肿瘤微循环的损伤及机理研究

肿瘤微循环对肿瘤生长和转移至关重要. 冷热交替治疗已被证实优于以往的单一冷疗或热疗, 其对于肿瘤微循环的损伤程度及机理是肿瘤成功治疗的关键, 但目前该方向的研究甚少. 利用裸鼠脊背皮翼肿瘤视窗模型结合激光共聚焦显微镜技术综合比较了单冷、单热及冷热交替对肿瘤微循环的损伤程度, 并建立了分析肿瘤血管在热物理作用下受力的理论模型, 通过数值计算获得了冷热交替治疗过程中肿瘤微循环血管壁的受力大小. 研究结果发现, 冷热交替造成了肿瘤微循环尤其是肿瘤中央成熟血管严重的结构性损伤. 冷热交替过程中, 血管壁先后受到迅速变化的方向相反的热应力, 有可能在血管壁形成微小裂纹. 随后加热造成的血流快速再灌注对血管壁的作用力存在应力集中现象, 推测血液流动对肿瘤血管壁的快速冲击力很可能是造成血管壁破裂的关键因素, 初步揭示了冷热交替治疗过程中肿瘤血管发生严重损伤的机理.