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Search Results: 1 - 10 of 52436 matches for " 樊廷俊 "
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对虾非特异性免疫与对虾疾病监控的研究进展

海洋科学 , 2002,
Abstract:
牙鲆鳃细胞与中国对虾淋巴细胞杂交细胞的体外培养

水产学报 , 2001,
Abstract: 利用pcg介导的细胞融合方法,将已成系的牙鲆鳃细胞与中国对虾淋马细胞进行细胞杂交,对所获得的杂交细胞用mem:mps(1:1)混合培养液进行体外培养,培养5天后发现,杂交细胞长成了单层,传代培养后仍可继续生长和分裂,通过逐渐提高混合培养液中mps的所占比例所筛选出的杂交细胞,目前已传12代,生长和分裂势头仍然十分旺盛,通过接种对虾杆现毒发现,10天后杂交细胞出现了明显的病变特征(细胞收缩变团圆,脱落,死亡),至14天细胞使几乎完全脱臂死亡,取病变细胞的培养上清0.3ml再加入到一瓶已长成单层的杂交细胞中,结果5天后杂交细胞也出出了明显的病变特和下,而未接种病毒的对照组杂交细胞生长正常,可禄步认为我们所得到的杂交细胞确为对虾(杆状)病毒的靶细胞。
THE MODE OF ACTION ON VITELLINE ENVELOPE OF Xenopus HATCHING ENZYME MOLECULES
非洲爪蟾孵化酶分子对卵黄膜作用方式的研究

,片桐千明
动物学研究 , 1998,
Abstract: The medium in which prehatching Xenopus laevis embryos were cultured (hatching medium) can solubilize vitelline envelope (VE) of dejellied uterine eggs and dimethyl casein. Western blotting analyses with antibodies against hatching enzyme revealed the presence of 60kD,and occasionally 40kD molecules in hatching medium. Results on VE solubilizing activity assay indicated that the fractions containing 60kD molecules exhibited strong solubilizing activity but those containing 40kD alone did not, although both fractions exhibited the same level of proteolytic activities. However, solubilization of VE was obtained when the 40kD fractions were mixed with an extremely low concentration of 60kD fractions that can not solubilize VE by itself, or were applied to the VE that had been pretreated with a low concentration of 60kD fractions. We proposed that recognition and/or processing of VE by CUB repeats in 60kD molecules made VE's solubilization possible by 40kD molecules.
鱼类精子冷冻保存的研究进展
汪小锋,
海洋科学 , 2003,
Abstract:
中国对虾酚氧化酶的部分生物化学特性的初步研究
汪小锋,
海洋科学 , 2003,
Abstract:
中国对虾淋巴组织培养细胞中立克次氏体增殖的形态学研究
汪岷,姜明,,刘晓云,汪晓峰,徐怀恕
海洋科学 , 2000,
Abstract:
硬骨鱼免疫系统的组成与免疫应答机制研究进展
王卫卫,吴谡琦,孙修勤,
海洋科学进展 , 2010,
Abstract:
非洲爪蟾孵化酶的cdna结构及其部分生物化学特性的研究
片桐千明 箕田康一 安增茂树
生物工程学报 , 1998,
Abstract: 以uvs.2为探针从第25期非洲爪蟾胚胎头背部的cdna文库中筛选出了一个1.8kb的孵化酶基因(xhe),其转录产物最早出现于第17期胚胎的头背部,在第30期转录量达到高峰,随后便逐渐减少。该基因含有编码514个氨基酸的一个开框阅读框架,含有信号肽和原酶序列。所推测出的成熟蛋白有425个氨基酸,包括位于n一端的含有200个氨基酸的金属蛋白酶序列和位于c端的两个各110个氨基酸的cub重复区。而uvs.2只代表该基因c端大约3/4的部分。同时还发现该酶分子量为60kda,是一种胰蛋白酶类型的金属蛋白酶。它很不稳定,在纯化过程中极易降解为40kda分子。60kda分子具有很强的卵黄膜溶解活性和蛋白酶活性。其中cub重复区很可能在介导卵黄膜和40kda分子中起着重要作用,而40kda分子很可能是在纯化操作过程中,由60kda分子发生降解或自身降解丢失了两个cub重复区而形成的,它只是60kda分子中的一个金属蛋白酶主功能区,所以它没有卵黄膜溶解活性,尽管仍具有很强的蛋白酶活性。
体外培养人角膜内皮细胞的UVB损伤及抗坏血酸的抗氧化保护作用研究
UVB damage and protection effect of ascorbic acid on human corneal endothelial cells in vitro

温茜,
WEN Qian
, FAN Ting-jun

- , 2015, DOI: 10.6040/j.issn.1671-9352.0.2015.197
Abstract: 摘要: 以非转染人角膜内皮(HCE)细胞系为体外实验模型系统研究了UVB氧化损伤、Asc抗氧化保护及其分子机理。体外培养的HCE细胞经UVB和/或Asc处理后,利用MTT和光镜对细胞的活力和形态进行了检测,利用8-羟基脱氧鸟苷免疫荧光染色对DNA的氧化损伤进行了检测,并利用二氢乙啶染色对胞内活性氧(ROS)的水平进行了检测。结果显示,100~800 mj/cm2的UVB辐射能剂量和时间依赖性地损伤HCE细胞的活力;200 mj/cm2 UVB(自然太阳光中的平均辐射剂量)能引起HCE细胞发生皱缩,并显著增加细胞的DNA氧化损伤程度及胞内ROS水平;而1 mmol/L Asc不仅能显著增强HCE细胞的活力、促进细胞分裂,而且还能显著降低200 mj/cm2 UVB所引起的DNA氧化损伤及胞内ROS水平。综上所述,UVB通过诱导ROS的产生进而引起DNA氧化损伤,对HCE细胞具有显著的氧化损伤作用;而Asc能够通过降低UVB诱导的ROS水平进而保护DNA免受氧化损伤,对HCE细胞的UVB损伤具有一定的抗氧化保护作用。本文研究结果对于利用Asc等抗氧化保护剂保护HCE细胞免受UVB氧化损伤具有一定的理论指导价值。
Abstract: To postulate the damage effect of UVB and the protection effect of antioxidant ascorbic acid(Asc), the oxidantive effect of UVB, the protective effect of Asc, and the related molecular mechanisms were investigated systematically using an in vitro model of non-transfected human corneal endothelial(HCE) cells in this study. After HCE cells were exposed to UVB irradiation and/or Asc, the viability and morphology were examined by MTT assay and light microscopy, DNA oxidative damage was detected by 8-hydroxydeoxyguanosine immunofluorescent staining, and the level of reactive oxygen species(ROS) was assayed by dihydroethidium staining. Our results showed that 100~800 mj/cm2 UVB irradiation could decline the viability of HCE cells in a dose-and time-dependent manner. UVB at 200 mj/cm2(the average dose of UVB in natural sunlight) could induce cell shrinkage, oxidative damage of DNA, and elevation of ROS level significantly. Whereas, 1 mmol/L Asc, with significant enhancing effects on the viability and proliferation of HCE cells, could significantly reduce the oxidative damage of DNA and the ROS level in 200 mj/cm2 UVB-irradiated HCE cells. In conclusion, UVB irradiation has a significant damaging effect on HCE cells by inducing DNA oxidative damage via ROS generation, while Asc has an obvious protecting effect on UVB-irradiated HCE cells by eleminatingDNA oxidative damage via inhibiting ROS generation. These findings can be used as certain theory guidance in protecting HCE cells from UVB-induced oxidative damage by employing Asc and the other antioxidants
一种新型脱细胞猪角膜基质载体支架的制备及其鉴定研究
Preparation and characterization of a novel acellular porcine cornea stromata carrier scaffold

,单鸣,庞鑫
FAN Ting-jun
, SHAN Ming, PANG Xin

- , 2017, DOI: 10.6040/j.issn.1671-9352.0.2017.112
Abstract: 摘要: 为了获得基于异种角膜材料的理想组织工程角膜载体支架,首次建立了脱氧胆酸钠(SD)和原钒酸钠(SO)联合处理的技术方法,利用新鲜猪角膜进行了脱细胞角膜基质(aPCS)的制备及其鉴定研究。用角膜板层刀从新鲜猪角膜中切下厚度450 μm的角膜片,分别选用SD联合SO、十二烷基磺酸钠(SDS)和Triton X-100的去细胞处理方法制备出SD-aPCS、SDS-aPCS和Triton-aPCS共3种支架,利用外观照相、分光光度计、石蜡切片苏木紫-伊红(HE)染色、冰冻切片DAPI和阿利新蓝染色检测了其理化性质和组织结构;对SD-aPCS进行扫描和透射电镜鉴定后,进而利用噻唑蓝(MTT)、石蜡切片HE染色、冰冻切片DiI荧光观察以及免疫荧光细胞化学染色评估了其对非转染人角膜基质(ntHCS)细胞的毒性与生物相容性。检测结果发现,3种aPCS的细胞脱除干净且在干重和含水量上没有显著差异;其中,SD-aPCS的透明性与糖胺聚糖(GAG)含量最高、SDS-aPCS次之、Triton-aPCS最低;除Triton-aPCS的组织结构出现了明显的紊乱外,SD-aPCS和SDS-aPCS的组织结构均排列规则。在电镜下,SD-aPCS的前弹力层表面平整、无裂痕,板层结构和胶原纤维超微结构正常;此外,SD-aPCS浸提液对ntHCS细胞没有毒性作用,注射接种到SD-aPCS支架内的ntHCS细胞与支架嵌合紧密,随体外培养时间的延长而逐渐伸展和迁移,且细胞仍保持有其固有标志蛋白—波形蛋白,细胞连接蛋白—间隙连接蛋白-43和整联蛋白,以及膜运输蛋白—钠钾泵的阳性表达。由此可见,利用SD联合SO的方法所制备SD-aPCS具有理想的理化性质、组织结构和生物相容性,可作为一种理想的载体支架用于组织工程角膜的体外构建及其相关应用研究。
Abstract: To obtain a satisfied carrier scaffold from xenogeneic corneal stroma for corneal tissue engineering, a novel technique using detergents of sodium deoxycholate(SD)and sodium orthovanadate(SO)was established to prepare an acellular porcine cornea stromata(aPCS)scaffold from fresh porcine cornea, and characterize its essential property for the first time in this study. The anterior lamella, 450 mm in thickness and sliced off from each cornea of fresh porcine eyeballs using a microkeratome, was decellularized with SD combined with SO, sodium dodecyl sulfate(SDS), and Triton X-100, and three kinds of aPCS scaffolds, ie SD-aPCS, SDS-aPCS and Triton-aPCS, were obtained, respectively. The physichemical property and histological structure of them was evaluated by light microscopy, spectrophotometer, paraffin section with hematoxylin-eosin(HE)staining, and frozen section with DAPI staining, and alcian blue staining, respectively. The ultrastructure of the SD-aPCS scaffold was verified by scanning electronic microscopy(SEM) 山 东 大 学 学 报 (理 学 版)第52卷 - 第5期樊廷俊,等:一种新型脱细胞猪角膜基质载体支架的制备及其鉴定研究 \=-and transmission electron microscopy(TEM), and their cytotoxicity and biocompatibility to non-transfected human corneal stromal(ntHCS)cells was evaluated by methyl thiazolyl tetrazolium(MTT), HE staining, DiI-fluorescence observation, and immunocytofluorescent staining. The results showed that the dry weight and water content of the three kinds of aPCS scaffolds had no significant difference, their innate cells were removed, and all of them were completely decellularized. Among them, the SD-aPCS scaffold had the highest transparency and glycosaminoglycan(GAG)content, followed by the SDS-aPCS and Triton-aPCS scaffold successively. In addition to the obvious disorder of the Triton-aPCS scaffold, the histological
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