Mesenchymal stem cells as a gene therapy carrier for treatment of fibrosarcoma

Background aimsCell-based gene therapy is an alternative to viral and non-viral gene therapy. Emerging evidence suggests that mesenchymal stem cells (MSC) are able to migrate to sites of tissue injury and have immunosuppressive properties that may be useful in targeted gene therapy for sustained specific tissue engraftment.MethodsIn this study, we injected intravenously (i.v.) 1 × 10⁶ MSC, isolated from green fluorescent protein (GFP) transgenic rats, into Rif-1 fibrosarcoma-bearing C3H/HeN mice. The MSC had been infected using a lentiviral vector to express stably the luciferase reporter gene (MSC-GFP-luci). An in vivo imaging system (IVIS 200) and Western blotting techniques were used to detect the distribution of MSC-GFP-luci in tumor-bearing animals.ResultsWe observed that xenogenic MSC selectively migrated to the tumor site, proliferated and expressed the exogenous gene in subcutaneous fibrosarcoma transplants. No MSC distribution was detected in other organs, such as the liver, spleen, colon and kidney. We further showed that the FGF2/FGFR pathways may play a role in the directional movement of MSC to the Rif-1 fibrosarcoma. We performed in vitro co-culture and in vivo tumor growth analysis, showing that MSC did not affect the proliferation of Rif-1 cells and fibrosarcoma growth compared with an untreated control group. Finally, we demonstrated that the xenogenic MSC stably expressing inducible nitric oxide synthase (iNOS) protein transferred by a lentivirus-based system had a significant inhibitory effect on the growth of Rif-1 tumors compared with MSC alone and the non-treatment control group.ConclusionsiNOS delivered by genetically modified iNOS-MSC showed a significant anti-tumor effect both in vitro and in vivo. MSC may be used as a target gene delivery vehicle for the treatment of fibrosarcoma and other tumors.     

Adiponectin 基因剔除LacZ基因敲入小鼠模型的建立

adiponectin是脂肪细胞特异分泌的一种活性蛋白质,具有增加胰岛素敏感性、抗炎及抗动脉硬化等活性.建立adiponectin基因剔除β-半乳糖苷酶基因(LacZ)敲入小鼠模型,可为整体动物水平研究adiponectin基因功能及其表达调控机制等提供理想工具.根据生物信息学方法获得adiponectin基因组序列,设计基因剔除及敲入策略,在adiponectin基因第2和第3号外显子剔除的同时,在其ATG和信号肽序列后顺接LacZ基因完整编码序列,构建完成了Adipo-LacZ-XpPNT基因剔除质粒.通过电穿孔将打靶质粒转入ES细胞,以G418和ganciclovir进行药物筛选,获得药物抗性的ES细胞克隆,PCR和DNA印迹鉴定出正确同源重组克隆.将同源重组的ES细胞克隆注入小鼠囊胚得到嵌合体小鼠,嵌合体小鼠与C57BL/6J小鼠交配产生杂合子小鼠,杂合子间交配获得adiponectin基因剔除LacZ基因敲入纯合子小鼠.经RT-PCR、RNA印迹和ELISA检测证实纯合子小鼠脂肪和血清中adiponectin基因表达呈阴性.RT-PCR、RNA印迹及蛋白质印迹检测发现,LacZ基因在突变小鼠脂肪组织中有特异性表达,其表达谱与内源性adiponectin基因的表达谱一致.但在脂肪组织及外周血中未能检测到LacZ活性,且血清中LacZ蛋白亦呈阴性.由此成功建立了adiponectin基因完全灭活及LacZ基因以内源性adiponectin基因表达谱表达的小鼠模型,为进一步研究该基因功能及其表达调控创造了有利条件.     

三种夜蛾成虫口器感器的超微形态

为确定不同种类夜蛾口器及其感器在超微结构上的差异, 采用扫描电子显微镜对棉铃虫Helicoverpa armigera (Hübner)、 烟夜蛾H. assulta (Guenée)和银纹夜蛾Argyrogramma agnata (Staudinger)3种鳞翅目(Lepidoptera)夜蛾科(Noctuidae)重要农业害虫雌、 雄成虫口器感器的超微形态进行了观察和比较。结果表明： 3种夜蛾雌、 雄成虫口器感器类型均无明显差异。棉铃虫和烟夜蛾口器感器在类型和形状上十分类似, 均具有毛形、 锥形和栓锥形感器; 喙管末端的栓锥感器粗、 密, 棱纹明显。银纹夜蛾口器感器与两种铃夜蛾区别明显, 除上述3类感器外, 还具有腔锥形感器; 其喙管末端的栓锥感器细、 疏, 棱纹不明显。结果显示口器感器可用于夜蛾的分类及亲缘关系研究。     

祁连山高寒草地甘肃臭草叶性状与坡向间的关系

植物功能性状可反映植物对环境的适应。在祁连山高寒退化草地, 利用ArcGIS建立研究区域的数字高程模型, 提取坡向数据, 采用标准化主轴估计方法(standardized major axis estimation, SMA), 研究了不同坡向甘肃臭草(Melica przewalskyi)叶性状间的关系。结果表明: 1)甘肃臭草叶面积和叶体积在北坡最大, 东坡、西坡、南坡逐渐减小, 叶干质量无显著变化; 2)叶面积与叶干质量在北坡、东坡和西坡呈等速生长关系, 在南坡呈异速生长关系, 且叶干质量的增长速度大于叶面积的增长速度; 3)叶体积与叶干质量在4个坡向上呈异速生长关系, 且叶干质量的增长速度均小于叶体积的增长速度。甘肃臭草叶性状关系随坡向的变化反映了该物种在异质生境中具有较强的叶片形态可塑性, 从而有利于其适应和占据高寒退化生境。     

Distribution and diversity of Gallionella-like neutrophilic iron oxidizers in a tidal freshwater marsh

Microbial iron oxidation is an integral part of the iron redox cycle in wetlands. Nonetheless, relatively little is known about the composition and ecology of iron-oxidizing communities in the soils and sediments of wetlands. In this study, sediment cores were collected across a freshwater tidal marsh in order to characterize the iron-oxidizing bacteria (FeOB) and to link their distributions to the geochemical properties of the sediments. We applied recently designed 16S rRNA primers targeting Gallionella-related FeOB by using a nested PCR-denaturing gradient gel electrophoresis (DGGE) approach combined with a novel quantitative PCR (qPCR) assay. Gallionella-related FeOB were detected in most of the samples. The diversity and abundance of the putative FeOB were generally higher in the upper 5 to 12 cm of sediment than in deeper sediment and higher in samples collected in April than in those collected in July and October. Oxygen supply by macrofauna appears to be a major force in controlling the spatial and temporal variations in FeOB communities. The higher abundance of Gallionella-related FeOB in April coincided with elevated concentrations of extractable Fe(III) in the sediments. Despite this coincidence, the distributions of FeOB did not exhibit a simple relationship to the redox zonation inferred from the geochemical depth profiles.     

广东省蛇类新纪录——越南烙铁头蛇

2009年6月,在广东省信宜市云开山自然保护区调查时采集到越南烙铁头蛇(Ovophis tonkinensis)标本2号,为广东省新纪录,为该种在中国分布的有效性提供了重要的依据.标本保存在华南濒危动物研究所.     

Metabolic profiling of strawberry (Fragaria x ananassa Duch.) during fruit development and maturation

Strawberry (Fragaria × ananassa Duch), a fruit of economic and nutritional importance, is also a model species for fleshy fruits and genomics in Rosaceae. Strawberry fruit quality at different harvest stages is a function of the fruit's metabolite content, which results from physiological changes during fruit growth and ripening. In order to investigate strawberry fruit development, untargeted (GC-MS) and targeted (HPLC) metabolic profiling analyses were conducted. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were employed to explore the non-polar and polar metabolite profiles from fruit samples at seven developmental stages. Different cluster patterns and a broad range of metabolites that exerted influence on cluster formation of metabolite profiles were observed. Significant changes in metabolite levels were found in both fruits turning red and fruits over-ripening in comparison with red-ripening fruits. The levels of free amino acids decreased gradually before the red-ripening stage, but increased significantly in the over-ripening stage. Metabolite correlation and network analysis revealed the interdependencies of individual metabolites and metabolic pathways. Activities of several metabolic pathways, including ester biosynthesis, the tricarboxylic acid cycle, the shikimate pathway, and amino acid metabolism, shifted during fruit growth and ripening. These results not only confirmed published metabolic data but also revealed new insights into strawberry fruit composition and metabolite changes, thus demonstrating the value of metabolomics as a functional genomics tool in characterizing the mechanism of fruit quality formation, a key developmental stage in most economically important fruit crops.     

The absence of muscle segment homeobox 2 leads to the pyroptosis of ameloblasts by inducing squamous epithelial hyperplasia in the enamel organ

Muscle segment homeobox 2 (MSX2) has been confirmed to be involved in the regulation of early tooth development. However, the role of MSX2 has not been fully elucidated in enamel development. To research the functions of MSX2 in enamel formation, we used a Msx2^−/− (KO) mouse model with no full Msx2 gene. In the present study, the dental appearance and enamel microstructure were detected by scanning electron microscopy and micro-computed tomography. The results showed that the absence of Msx2 resulted in enamel defects, leading to severe tooth wear in KO mice. To further investigate the mechanism behind the phenotype, we performed detailed histological analyses of the enamel organ in KO mice. We discovered that ameloblasts without Msx2 could secrete a small amount of enamel matrix protein in the early stage. However, the enamel epithelium occurred squamous epithelial hyperplasia and partial keratinization in the enamel organ during subsequent developmental stages. Ameloblasts depolarized and underwent pyroptosis. Overall, during the development of enamel, MSX2 affects the formation of enamel by regulating the function of epithelial cells in the enamel organ.     

Hyperpolization-activated Ca channels in guard cell plasma membrane are involved in extracellular ATP-promoted stomatal opening in Vicia faba

Extracellular ATP (eATP) plays essential roles in plant growth, development, and stress tolerance. Extracellular ATP-regulated stomatal movement of Arabidopsis thaliana has been reported. Here, ATP was found to promote stomatal opening of Vicia faba in a dose-dependent manner. Three weakly hydrolysable ATP analogs (adenosine 5′-O-(3-thio) triphosphate (ATPγS), 3′-O-(4-benzoyl) benzoyl adenosine 5′-triphosphate (Bz-ATP) and 2-methylthio-adenosine 5′-triphosphate (2meATP)) showed similar effects, indicating that ATP acts as a signal molecule rather than an energy charger. ADP promoted stomatal opening, while AMP and adenosine did not affect stomatal movement. An ATP-promoted stomatal opening was blocked by the NADPH oxidase inhibitor diphenylene iodonium (DPI), the reductant dithiothreitol (DTT) or the Ca²⁺ channel blockers GdCl₃ and LaCl₃. A hyperpolarization-activated Ca²⁺ channel was detected in plasma membrane of guard cell protoplast. Extracellular ATP and weakly hydrolyzable ATP analogs activated this Ca²⁺ channel significantly. Extracellular ATP-promoted Ca²⁺ channel activation was markedly inhibited by DPI or DTT. These results indicated that eATP may promote stomatal opening via reactive oxygen species that regulate guard cell plasma membrane Ca²⁺ channels.     

体重指数在评估内科住院患者肺心病发生中的价值

目的:探讨体重指数在预测内科住院患者肺心病发生中的价值。方法:利用随机数表随机入选解放军第十二医院2012年1月~2012年12月间内科住院患者92例,其中男性47例,女性45例,年龄在22岁~87岁之间。体重指数(Body mass index,BMI)应用公式:BMI=体重/身高2(kg/m2)计算。用酶法检测血清总胆固醇、甘油三酯;葡萄糖氧化酶法测定空腹血糖。结果:与非肺心病组患者比较,肺心病组患者体重指数明显升高(38.96±5.77 vs 29.72±7.879,P0.01);肺心病组患者肺性脑病病史的比率明显升高(26.7%vs 0%,P0.01)。肥胖和极度肥胖的患者合并肺心病病史的比率分别为53.3%、46.7%。多分类Logistic回归分析显示,体重指数与肺心病密切相关。ROC曲线分析显示,体重指数预测肺心病的临界值为BMI32 kg/m2,预测肺心病的ROC曲线下面积为0.860,ROC曲线下面积大于0.7,预测价值较高。Youden指数为0.596,体重指数预测最佳临界值为BMI大于32,诊断特异性为66.2%,敏感性为93.3%,阳性预测值为34.96%,阴性预测值为98.07%。结论:内科住院患者体重指数与肺心病发病率密切相关,体重指数大于32 kg/m2预测内科住院患者肺心病发病有较高的价值。