鸭梨果实石细胞分化特性研究

以鸭梨为试材,利用透射电镜显微技术和组织化学技术对梨果实石细胞分化及次生壁形成过程进行研究。结果表明,石细胞分化始于开花第7天,原生质体收缩于细胞一侧,随后胞内出现颗粒状凝聚物,次生壁逐渐增厚,之后胞质继续收缩消失,早期形成的石细胞多以群体形式同时出现。石细胞次生壁形成是逐步进行的,即先形成纤维素或半纤维素网架结构,再进行木质化填充。透射电镜下石细胞分化过程中依次呈现出细胞核染色质凝聚、细胞核变形、核膜界限不清晰的现象;胞内出现同心层次的自噬泡,形成大量囊泡;细胞核消失,内质网断裂并大量增生,质膜向内突起形成泡状结构;次生壁增厚,胞内细胞器仅有线粒体和内质网,最终细胞质收缩消失。     

Identification of Suspected Stem/progenitor Cells in Bovine Mammary Epithelial Cells Culture System

To develop the potential function of dairy cow mammary stem cells (DCMECs) in regulation of lactation,we identify putative DCMECs which were BrdU label retaining epithelial cells,at the same time,analysis the location of two new mammary stem cells molecular marks FNDC3B and PROCR to verify the feasibility of them to indicate DCMECs.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-1 and their receptors were detected along with cell passage by Real-time quantitative PCR.The results showed that the proportion of BrdU label-retaining epithelial cells was nearly 0.4% after 25 d continuous culture (passaged 4 times) and few cells were positive for FNDC3B or PROCR.Moreover,we observed the BrdU labelled epithelial cells by asymmetric division.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-Ⅰ and their receptors in primary and passage cells were extremely significant difference(P<0.01).DCMECs would rapidly lose some physiological characteristics and the ability of milk synthesis when not under the condition of induction of lactation differentiation,but a certain percentage of mammary stem/progenitor cells will be retained,whose potential effects on the regulation of lactation and mammary acinar remodeling were worthy of attention.     

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.     

Interaction of NR2D subunit of NMDA receptor with MOCA

AIM: To identify the potential proteins interacting with NR2D subunit of NMDA receptor by yeast two-hybrid screening and to investigate the role of NR2D in excitotoxicity of the retina.METHODS: The Clontech GAL4 yeast two-hybrid system was used to screen the mouse brain cDNA library, and the bait plasmid containing C-terminus of NR2D was constructed. Physical interaction between 2 proteins was verified by co-immunoprecipitation assay. The subcellular localization of 2 proteins in the mouse retina was observed under microscope with immunofluorescence.RESULTS: Modifier of cell adhesion (MOCA) was identified as a new protein interacting with NR2D. MOCA and NR2D were co-expressed in the mouse retina. CONCLUSION: MOCA specifically interacts with NR2D, which provides the experimental basis for identifying the role of glutamate excitotoxicity in the retina neurodegeneration.     

Functional analysis of the venom allergen-like protein gene from pine wood nematode Bursaphelenchus xylophilus using a baculovirus expression system

The pine wood nematode, Bursaphelenchus xylophilus, infects pine trees, leading to fatal pine wilt disease. Here, recombinant venom allergen-like protein (VAP) was obtained by expressing Bx-vap-1 in insect cells. Three-year-old Pinus massoniana were inoculated with recombinant VAP, simulating B. xylophilus esophageal gland secretions. Recombinant VAP up-regulated α-pinene synthase gene expression, the trees showed disease symptoms 15 d after inoculation and the xylem pith revealed brown tissue discoloration, indicating that recombinant VAP could damage P. massoniana cells. Recombinant VAP did not, however, lead to cavitation, indicating that the VAP secreted from B. xylophilus acts as a defense response elicitor.     

某厂区办公楼的结构设计

笔者结合工程实例,采用SATWE软件对单跨框架结构进行计算分析,并浅析了设计中应重点注意的问题。     

In vitro effect of levamisole on the cell viability,phagocytosis and respiratory burst of Barbel chub (Squaliobarbus curriculus) macrophages

The present study was to determine in vitro effect of levamisole on the immune functions of Barbel chub (Squaliobarbus curriculus), head–kidney‐derived (HKM), spleen‐derived (SM) and peripheral blood monocyte‐derived macrophage (PBM). Macrophages were incubated with levamisole 10³, 10¹, 10⁻¹ and 10⁻³ ng mL⁻¹ to assay the cell viability, respiratory burst and phagocytosis. The results showed that macrophages treated with levamisole 10⁻³ ng mL⁻¹ gave a maximum respiratory burst response, whereas levamisole 10³ ng mL⁻¹ had no effect. Phagocytosis activity of the macrophages treated with levamisole enhanced significantly when compared to control, maximum response being at 10⁻³ ng mL⁻¹. While using the methylthiazoletetrazolium method to measure the cell activity for 12, 24, 36, 48, 60 h, there was no significant role on proliferation and the cell viability began to decline after 24 h. In conclusion, levamisole is a potent enhancer of Barbel chub macrophage activity with low concentration.     

Vitamin C enhanced myocardial differentiation of dedifferentiated fat cells

AIM: In order to observe the myocardial differentiation capacity of the dedifferentiated fat (DFAT) cells treated with vitamin C in vitro. METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture. The DFAT cells of passage 3 were used in the study. Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks. The cell morphology was observed under microscope. The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot. RESULTS: Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture. The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence. After treated with neonatal rat heart cell lysate, the DFAT cells became cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure. The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells. The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C. No spontaneous beating cell was observed. CONCLUSION: Vitamin C enhances the differentiation of DFAT cells into cardiomyocyte-like cells.     

浅谈地下车站侧墙大块定型移动模架施工技术

随着城市地铁的飞速发展,地铁车站在条件满足的情况下大都采用明挖法施工。在施工主体侧墙时采用满堂红支架法施工,但因施工时工期较长且易存在一些质量通病,因此,近年来大块定型移动模架被得到了广泛的应用。