Histological and immunohistochemical evaluation of granulosa cells during different stages of folliculogenesis in bovine ovaries          下载免费PDF全文

APP Teh  UZ Izzati  K Mori  N Fuke  T Hirai  G Kitahara  R Yamaguchi 《Reproduction in domestic animals》2018,53(3):569-581

Bovine granulosa cells (GC) vary in their morphological aspect during different stages of folliculogenesis. In this study, 10 morphologically normal bovine ovaries were collected to study the structural aspects of different stages of GC using intermediate filament protein antibodies including cytokeratin AE1/AE3 (AE1/AE3), vimentin, nectin‐4 and desmin. Hormonal immunolocalization was assessed using the immunomarkers anti‐Müllerian hormone (AMH) and inhibin alpha. In addition, tumour markers and proliferation markers using c‐erbB‐2 oncoprotein and proliferating cell nuclear antigen, respectively, were investigated. The immunolabelling of AE1/AE3 in GC was strongest in the early follicle stage and gradually decreased when reaching the Graafian follicle stage. Its immunolabelling increased again as the stage progressed from stage I to stage III. The immunolabelling of inhibin alpha was inversely proportional to that of AE1/AE3 in the developing ovarian follicles as their immunolabelling is opposite to each other during folliculogenesis. AMH was immunopositive in almost all GC stages in different intensities and percentages, except for some negative staining in the atretic IV follicles. The atretic IV follicle is a unique type of atretic follicle that shows Call‐Exner body formation, which was mainly found in older cows in this study. The distinct patterns of immunoreactivity for various types of immunomarkers in the different GC stages will play an important role in diagnostic assistance of various follicle conditions, including cystic ovaries and GC tumours.  相似文献   

Effects of HIF-1α silencing on proliferation of rat hepatoma cells     

WU Yu-dan  NI Jia-yan  CHEN Yao-ting  SUN Hong-liang  XU Lin-feng 《园艺学报》2013,29(7):1207-1212

AIM：To study the effect of hypoxia-inducible factor 1&alpha; (HIF-1&alpha;) silencing on the proliferation of hepatoma cells under hypoxia. METHODS：Rat hepatoma cell line CBRH-7919 was used in this study. Hypoxia model was established by treating the cells with cobalt chloride (CoCl2). The expression of HIF-1&alpha; was silenced by small interfe-rence RNA. Real-time RT-PCR and Western blotting were used to detect the mRNA and/or protein expression of HIF-1&alpha;, vascular endothelial growth factor (VEGF), p21 and cyclin D1 in CBRH-7919 cells under hypoxia. The proliferation of CBRH-7919 cells was measured by the technique of 5-bromo-2’-deoxyuridine (BrdU) incorporation. RESULTS：The expression of HIF-1&alpha; and VEGF at mRNA and protein levels was significantly increased under hypoxia (P<0.05). Silencing of HIF-1&alpha; significantly inhibited the expression of HIF-1&alpha;, VEGF and cyclin D1 at mRNA and/or protein levels, while increased the protein expression of p21 (P<0.05). The BrdU-positive cells in HIF-1&alpha; siRNA transfection group were significantly less than those in control group. CONCLUSION：HIF-1&alpha; silencing significantly inhibits the proliferation of hepatoma cells under hypoxia.  相似文献   

Impact of alpha-cypermethrin on honey bees foraging on spring oilseed rape (Brassica napus) flowers in field conditions     

Karise R  Viik E  Mänd M 《Pest management science》2007,63(11):1085-1089

BACKGROUND: Cruciferous oil-bearing crops have gained in importance worldwide. The expansion of the growing area of these crops has caused a proliferation of pests. Exposure to organophosphate, carbamate and pyrethroid insecticides has been associated with bee poisoning in food crops. This study examines the repellent effect of alpha-cypermethrin on the number of foraging honey bees, Apis mellifera L., on fields of spring oilseed rape, Brassica napus L. var. oleifera. RESULTS: The first experiment was conducted on differently sprayed 10 m(2) experimental plots where alpha-cypermethrin was applied at different times. Another experiment was conducted on a 4 ha seed production field divided into two parts: one part was treated with alpha-cypermethrin and the other was not treated with this insecticide. The results show that there was no difference in the number of honey bees between alpha-cypermethrin-treated and untreated patches. The result persisted through three observation years, regardless of varying flower and honey bee densities. CONCLUSION: No repellent effect of the insecticide on honey bees was found even 24 h after spraying. The density of oilseed rape flowers most likely played a major role in choosing the foraging area.  相似文献   

Evaluation of sedation for standing clinical procedures in horses using detomidine combined with buprenorphine     

Polly Taylor  Karen Coumbe  Frances Henson  David Scott  Alan Taylor 《Veterinary anaesthesia and analgesia》2014,41(1):14-24

  相似文献   

浙江凤阳山-百山祖国家级自然保护区植物群落ALPHA多样性特征     

谭毓佳  袁留斌  陈德良  赵鹂  谭珊珊  吴香花  顾少敏 《浙江林业科技》2014,(3)

采用样方调查法对浙江凤阳山--百山祖国家级自然保护区内的45个样地的物种组成、alpha多样性进行分析。结果表明：各样地乔、灌、草3个层次的Shannon-Wiener指数（H）、Simpson指数（D）和Pielous均匀度指数（Jsw）以五岭坑保护站的米槠（Castanopsis carlesii）、毛锥（Castanopsis fordii）林最高,茶木淤保护站的金钱松（Pseudolarix amabilis）林样地最低;各植物群落类型乔木层的物种丰富度以常绿阔叶林最高;灌木层以针阔叶混交林的物种数最多,草甸样地的灌木物种数最少;草本层则以竹林物种数最为丰富;乔木层各群落的 H和D均以常绿阔叶林最高;灌木层的H以针阔叶混交林最高,草本层中各植物群落类型的H、D、Jsw均以竹林最高;在百山祖自然保护区中,常绿阔叶林可以维持较高的木本植物物种多样性,而竹林出现明显纯林化现象,应加以控制。  相似文献   

三黄鸡a干扰素的克隆、表达及抗病毒活性初步研究     

李秋霞  刘永录  龚剑  马仲彬  陈乃壮  杨明锋  张国祖 《动物医学进展》2011,(10):63-66

采用PCR方法从三黄鸡血液基因组中扩增鸡α干扰素全基因,并克隆和测序.序列分析表明,基因全长为582 bp,亚克隆其成熟蛋白编码基因489 bp.将该片段与表达载体pET-28a连接克隆至大肠埃希菌DH5α菌株,经测序和酶切鉴定,选取正向插入、读码框正确的阳性克隆.构建重组质粒并转化BL21(DE3),经IPTG诱导,...  相似文献   

Selection for contrasting tocopherol content and profile in Ethiopian mustard     

Leonardo Velasco  Elena García‐Navarro  Begoña Pérez‐Vich  José M. Fernández‐Martínez 《Plant Breeding》2013,132(6):694-700

Tocopherols are compounds present in oilseeds with major effect on nutritional and technological properties of seed oils. Brassica spp. mainly contain varying concentrations of α‐tocopherol, with high vitamin E value, and γ‐tocopherol, which contributes to oil stability. The objective of this research was to select for contrasting tocopherol content and profile in Ethiopian mustard (Brassica carinata A. Braun). A collection of 211 F_4:5 lines was evaluated for tocopherol content and profile, and divergent selection was conducted till the S_3:4 generation. Selection resulted in two lines with increased tocopherol content (244 and 243 mg/kg seed as average of four environments), one line with reduced tocopherol content (117 mg/kg), two lines with reduced α/γ‐tocopherol ratio (0.22 and 0.28) and two lines with increased α/γ‐tocopherol ratio (3.74 and 3.34). Control line C‐101 averaged 196 mg/kg with α/γ‐tocopherol ratio of 1.10. Reduced α/γ‐tocopherol ratio was controlled by recessive alleles at a single locus. Seed to plant correlation was high (r = 0.92), which suggests the feasibility of selecting at the single‐seed level.  相似文献   

重组猪干扰素α1冻干粉针剂在猪体内的药代动力学研究     

赵俊  俞海洋  王兴满  李京培  胡涛  胡勇  苏世云  李超  鲁兴生 《中国预防兽医学报》2013,35(9)

为研究重组猪干扰素α1 (rPoIFN-α1)冻干粉针剂在猪体内的药代动力学特征,本研究将实验猪随机分为3组:分别静脉、肌肉和皮下注射rPoIFN-α1 1.0×107 IU,每组设立对照组,于注射后间隔不同时间采血,采用细胞病变抑制法测定其血清样品中rPoIFN-α1的效价.结果表明,肌肉注射组和皮下注射组药动学特征符合一室开放模型,呈一级动力学消除,血药峰时间(Tmax)分别为2.872±0.314 h和4.000±0.539 h,消除半衰期(T1/2)分别为6.236±0.551 h和6.644±0.751 h,血药峰浓度(Cmax)分别为3.980±2.875× 104 IU/L和1.080±0.986× 104 IU/L.静脉注射组符合二室开放模型,呈一级动力学消除,分布半衰期(T1/2α)为0.136±0.021 h,消除半衰期(T1/2β)为5.195±0.351 h,Cmax值为6.380±0.546× 104 IU/L.rPoIFN-α1静脉、肌肉和皮下注射组清除率分别为0.930±0.132 L/h、0.677±0.228 L/h和0.879±0.210 L/h.与静脉注射组相比,肌肉注射组的生物利用度为47.82％,皮下注射组的生物利用度为35.09％.本实验结果提示这3种rPoIFN-α1的注射方法在猪体内吸收、消除较快,具有线性动力学特征.rPoIFN-α1肌肉注射给药的生物利用度高于皮下注射组,消除半衰期比静脉注射组延长,可以作为替代静脉注射的一种方便和安全给药途径.  相似文献   

放牧和刈割对呼伦贝尔草甸草原物种多样性的影响   总被引：3，自引：1，他引：2  

黄振艳  王立柱  乌仁其其格  李杰  杨晓刚 《草业科学》2013,30(4):602-605

研究了内蒙古呼伦贝尔草甸草原贝加尔针茅（Stipa baicalensis)群落物种多样性对放牧和刈割的响应。结果表明,放牧条件下优势种贝加尔针茅的重要值高于刈割利用的,而样方内的物种数、Margalef指数、Shannon Wiener指数和Pielou指数均低于刈割利用,说明刈割利用较放牧增加了草原贝加尔针茅群落的&alpha;多样性。  相似文献   

印度南瓜延伸因子基因CmEF1a的克隆与分析     

朱海生  刘建汀  温文旭  李永平  王彬  陈敏氡  温庆放 《核农学报》2019,33(6):1096-1104

真核生物延伸因子(EF1a)是一种重要的内参基因,广泛应用于RT-qPCR中。为获得印度南瓜EF1a基因,开发合适的荧光定量PCR引物,本研究通过转录组测序和RT-PCR方法获得了1条长度为1 701 bp的cDNA,命名为CmEF1a,GeneBank登录号:MH310443。结果表明,CmEF1a包含1个ORF,大小为1 344 bp,编码447个氨基酸,理论分子大小约为49.30 kD, 蛋白质等电点为9.14。Wolf Psort分析发现, CmEF1a蛋白亚细胞定位于细胞质基质中;Motif Scan分析显示, CmEF1a蛋白质的氨基酸序列2~432位和322~430位分别为EF1结构域和EF1的C端保守结构域。同源性分析表明, CmEF1a基因编码的蛋白质与同为葫芦科的中国南瓜、甜瓜和黄瓜同源蛋白的相似性达到99%,具有高度的保守性。在此基础上设计了1对RT-qPCR引物, 该引物具有较高的特异性和扩增效率, 在印度南瓜不同组织和不同胁迫处理下均能稳定表达, 适合作为内参基因应用于印度南瓜基因表达研究。本研究结果为印度南瓜重要功能基因的表达分析及相关分子调控机制的研究奠定了一定的理论基础。  相似文献