In vivo matrix-guided human mesenchymal stem cells

Microfracture of subchondral bone results in intrinsic repair of cartilage defects. Stem or progenitor cells from bone marrow have been proposed to be involved in this regenerative process. Here, we demonstrate for the first time that mesenchymal stem (MS) cells can in fact be recovered from matrix material saturated with cells from bone marrow after microfracture. This also introduces a new technique for MS cell isolation during arthroscopic treatment. MS cells were phenotyped using specific cell surface antibodies. Differentiation of the MS cells into the adipogenic, chondrogenic and osteogenic lineage could be demonstrated by cultivation of MS cells as a monolayer, as micromass bodies or mesenchymal microspheres. This study demonstrates that MS cells can be attracted to a cartilage defect by guidance of a collagenous matrix after perforating subchondral bone. Protocols for application of MS cells in restoration of cartilage tissue include an initial invasive biopsy to obtain the MS cells and time-wasting in vitro proliferation and possibly differentiation of the cells before implantation. The new technique already includes attraction of MS cells to sites of cartilage defects and therefore may overcome the necessity of in vitro proliferation and differentiation of MS cells prior to transplantation. Received 3 November 2005; received after revision 15 December 2005; accepted 4 January 2006     

The protease-activated receptor-3 (PAR-3) can signal autonomously to induce interleukin-8 release

Protease-activated receptors (PARs) play a clear role in the burst of inflammatory reactions and immune responses. However, for PAR-3, the most elusive member of the PAR family, the functional role is still largely unclear. It has been claimed that PAR-3 does not signal autonomously, although the wide expression of human PAR-3 indicates its important physiological roles. We demonstrate that in HEK-293 cells, stably transfected with human PAR-3, thrombin induced calcium signaling, IL-8 gene expression and IL-8 release. We confirmed this finding using human lung epithelial and human astrocytoma cells that express endogenous PAR-3. Moreover, thrombin exposure of HEK-293 cells resulted in ERK1/2 activation coinciding with IL-8 release. The effects of thrombin were not dependent on PAR-1 activation, as confirmed by PAR-1 gene silencing. Thus, we propose that PAR-3 is able to signal autonomously to induce IL-8 release mediated by ERK1/2 phosphorylation, which contributes actively to inflammatory responses. Received 9 December 2007; received after revision 16 January 2008; accepted 18 January 2008     

Olfactory ensheathing cells are attracted to, and can endocytose, bacteria

Olfactory ensheathing cells (OECs) have been shown previously to express Toll-like receptors and to respond to bacteria by translocating nuclear factor-kappaB from the cytoplasm to the nucleus. In this study, we show that OECs extended significantly more pseudopodia when they were exposed to Escherichia coli than in the absence of bacteria (p=0.019). Co-immunoprecipitation showed that E. coli binding to OECs was mediated by Toll-like receptor 4. Lyso-Tracker, a fluorescent probe that accumulates selectively in lysosomes, and staining for type 1 lysosome-associated membrane proteins demonstrated that endocytosed FITC-conjugated E. coli were translocated to lysosomes. They appeared to be subsequently broken down, as shown by transmission electron microscopy. No obvious adherence to the membrane and less phagocytosis was observed when OECs were incubated with inert fluorescent microspheres. The ability of OECs to endocytose bacteria supports the notion that OECs play an innate immune function by protecting olfactory tissues from bacterial infection.     

接种密度对Vero细胞在微载体表面生长的影响

研究了接种密度对Ｖｅｒｏ细胞在微载体表面生长行为的影响,发现培养过程中所获得的细胞最高密度随接种密度的增加而有所提高,当细胞的表面接种密度高于１．０×１０＾５ｃｅｌｌｓ／ｍｇＭＣ时,因贴壁后细胞扩展受到限制而不利于进入对数生长期。就整个Ｖｅｒｏ细胞培养过程,适宜的表面接种密度为３×１０＾－４－７×１０＾４ｃｅｌｌｓ／ｍｇＭＣ,此时可以获得较高的比生长速率和细胞增殖倍数。     

外源性H2S通过调节β-位淀粉样前体蛋白裂解酶1表达对PC12细胞APP／Aβ代谢的影响

目的观察外源性硫化氢对嗜铬细胞瘤细胞β-位淀粉样前体蛋白裂解酶1（BACE1）的调节作用,进而探讨其对淀粉样前体蛋白／β-位淀粉样蛋白代谢途径的影响。方法用硫氢化钠作外源性H2s供体,实验设空白对照组、NaHs50μmol／L组、NaHS100μmol／L组和NariS200μmol／L组,按分组浓度处理PC12细胞24h后,RT-PCR和Western blot法检测细胞内BACE1 mRNA及蛋白表达,并用Western blot法继而检测APP代谢过程中关键蛋白APP、C99、C83表达变化,EusA法检测细胞培养液中AB40和AB42水平。结果NaHS在实验浓度范围内从基因与蛋白两个水平上呈剂量依赖性下调BACE1表达,并下调C99、Ap40和Ap42蛋白表达,上调C83蛋白,各NaHS组分别与对照组比较,差别均有统计学意义（P〈0．05）,而对APP蛋白表达没有影响,各组间比较差别无显著性（P〉0．05）。结论外源性H2s具有通过调节PC12细胞BACE1表达下调APP／Aβ代谢的作用。     

弱激光照射周氏新对虾心脏细胞的显微和超微结构观察

采用He-Ne激光照射周氏新对虾的心脏组织.组织学和生物统计学表明:He-Ne激光照射后,心脏组织具有明显异染色质的细胞数量有所增加.在同一照射强度的情况下,照射的时间越长,其心脏组织具有明显异染色质的细胞数量就越多;电镜观察结果表明:经过2 h照射立即取材的心肌细胞,细胞核形态发生变化,异染色质增加,线粒体出现空泡化、嵴断裂、水肿,粗面内质网出现扩张、颗粒脱落.     

太阳能集热型溶液再生器性能实验研究

采用CaCl2溶液、LiCl溶液、质量比1∶1的CaCl2和LiCl混合溶液,研究了空气入口温度、含湿量、空气流量、溶液流量、溶液进口温度、进口浓度对集热型再生器性能的影响.实验结果表明:较低的溶液进口浓度和空气入口含湿量以及较高的溶液进口温度能够增加再生量;而空气入口温度升高时,再生量仅略有增加;CaCl2溶液的再生性能最优,1∶1混合溶液次之,LiCl溶液的再生性能最差.     

X射线荧光光谱法用于CIGS薄膜太阳电池中吸收层的定量分析

介绍了X荧光薄膜分析法测试CIGS薄膜太阳电池中吸收层的4种元素的比例,并对其进行了分析和研究,此方法测量速度快,精确度高,对于制备高效率的CIGS太阳电池具有重要的指导意义。     

用原子力显微镜观察活海马区神经元三维超微结构

在液体环境中,利用原子力显微镜成功地对体外培养的大鼠海马区神经元进行了超微结构的成像研究。分别获取了活海马神经元胞体、轴突、树突以及生长锥等结构清晰的原子力显微图像,并对其三维结构进行了定量分析。为进一步利用原子力显微镜研究海马神经元打下了基础。