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1.
目前很多女性受到不孕不育、自然流产和出生缺陷等诸多生殖相关疾病的困扰,其主要原因为生殖细胞减数分裂异常和卵母细胞质量下降。雌激素不仅参与调控哺乳动物的发情和性行为,也参与调控卵母细胞的生长发育。雌激素受体作为类固醇激素受体的一种,在介导雌激素发挥基因组效应和非基因组效应中有着重要的作用。目前关于雌激素受体调控卵母细胞成熟的报道多集中于硬骨鱼类,特别是新型膜受体GPR30,而在哺乳动物方面的研究鲜有报道。因此作者介绍了雌激素核受体ERα、ERβ与新型膜受体GPR30的结构与分布,简述了膜受体GPR30的不同类型配体的研究进展,综述了雌激素受体在调控卵母细胞发育中的相互作用,发现雌激素的核受体与膜受体在卵母细胞成熟的过程中可能发挥不同的调控作用,并进一步阐述了雌激素及其受体在卵母细胞成熟过程中的潜在调控机制。  相似文献   
2.
为了探讨牦牛HSP27(heat shock proteins27, HSP27)基因在牦牛卵母细胞和早期孤雌激活胚胎中的表达情况及定位,本试验通过实时荧光定量PCR检测牦牛HSP27基因在未成熟卵母细胞和成熟卵母细胞以及孤雌激活不同时期胚胎中的相对表达量,用免疫荧光染色法检测HSP27蛋白在未成熟卵母细胞和成熟卵母细胞以及孤雌激活不同时期胚胎中的表达与定位情况。结果显示,HSP27基因在牦牛未成熟卵母细胞和成熟卵母细胞以及体外培养孤雌激活不同时期胚胎中均可表达,其中在未成熟卵母细胞中的表达量显著高于成熟卵母细胞;在孤雌激活胚胎中,2~4细胞期表达量最高,囊胚期胚胎表达量最低,随着胚胎的发育,HSP27的表达量呈现逐渐降低的趋势;在2~4细胞期和囊胚期胚胎中的表达与其他组表达存在显著差异性,在5~8细胞期和桑椹胚中的表达无显著差异。免疫荧光染色结果表明,HSP27蛋白在牦牛卵丘细胞和卵母细胞中均有表达,在牦牛孤雌激活2~4细胞期、5~8细胞期和桑椹期胚胎中主要表达于细胞核内和细胞质中,在囊胚期主要表达于细胞核中;而HSP27蛋白的表达水平与基因表达水平基本一致。本研究为后续研究HSP27基因在牦牛生殖生理过程中的作用机制提供了参考。  相似文献   
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4.
Kaempferol (KAE) is a natural flavonoid present in different plant species and exhibits anti‐inflammatory, antioxidant, and anticancer therapeutic properties. In the present study, we investigated the influence and underlying mechanisms of KAE supplementation on porcine oocytes during in vitro aging. The results show that KAE treatment can alleviate the aging‐related reduction of developmental competence. We observed that the blastocyst production rate in aged oocytes treated with 0.1 μM KAE was significantly higher than in untreated aging oocytes (36.78 ± 0.86% vs. 27.55 ± 2.60%, respectively, p < .05). The KAE‐treated aging oocytes had significantly reduced levels of reactive oxygen species (p < .05). Furthermore, the mRNA levels of the embryonic pluripotency‐related genes Oct4, NANOG, and ITGA5 were significantly increased in blastocysts derived from KAE‐treated oocytes (p < .05). During excessive oocyte culture, KAE treatment maintained the mitochondrial membrane potential and reduced apoptosis; however, this was not observed in untreated aging oocytes. In conclusion, our results suggest that KAE treatment can alleviate the aging of porcine oocytes by reducing oxidative stress and improving mitochondrial function.  相似文献   
5.
We investigated whether high‐quality in vitro matured (IVM) oocytes can be distinguished from poor ones based on the morphological changes after treatment with hyperosmotic medium containing 0.2 mol/L sucrose in pigs. We hypothesize that IVM oocytes maintaining round shape have higher quality than mis‐shapened oocytes following dehydration. Oocyte quality was verified by determining embryonic developmental competence using in vitro fertilization, nuclear transfer and parthenogenetic activation. In all cases, the round oocytes had greater (p < .05) developmental competence than that of mis‐shapened oocytes in terms of blastocyst rate and total cell number in blastocysts obtained after 6 days of in vitro culture. We also confirm that round aged oocytes are higher in quality than mis‐shapened aged oocytes. In an attempt to find out why high‐quality oocytes maintain a round shape whereas poorer oocytes become mis‐shapened following sucrose treatment, we examined the arrangement of actin microfilaments and microtubules. Abnormal organization of these cytoskeletal components was higher (< .05) in mis‐shapened oocytes compared to round oocytes after 52 hr of IVM. In conclusion, sucrose treatment helps selection of high‐quality oocytes, including aged oocytes, in pigs. Abnormal cytoskeleton arrangements partly explain for low developmental competence of mis‐shapened oocytes.  相似文献   
6.
试验旨在探究高浓度葡萄糖对猪卵母细胞体外成熟及早期胚胎发育能力的影响。取体外分离处于生发泡期的猪卵丘卵母细胞复合体(COCs),分为3个处理组。分别用含葡萄糖浓度为5.6 mmol/L(C组)、10 mmol/L(G-1组)、15 mmol/L(G-2组)的培养液,进行体外成熟(IVM)处理,42 h后观察,并统计卵丘细胞扩散情况和第一极体排出率;对体外成熟42 h后的卵母细胞孤雌激活,统计2-细胞、4-细胞和第7天囊胚发育。结果发现,G-1组和G-2组卵丘细胞扩散度显著低于C组(P<0.05);G-1组和G-2组的MII期卵母细胞死亡率和存活率与C组相比无显著差异(P>0.05),但G-1组极体率显著降低(P<0.05),G-2组极体率极显著低于C组(P<0.01)。孤雌激活后,与C组相比,G-1组和G-2组的2-细胞分裂率显著降低(P<0.05),4-细胞分裂率以及囊胚发育率均极显著降低(P<0.01),但G-1、G-2组囊胚细胞数量与C组相比无显著性差异(P>0.05)。进一步线粒体染色发现,G-1组和G-2组的线粒体与C组相比分布不均。与C组相比,荧光结果显示G-1组和G-2组不仅活性氧(ROS)水平极显著升高(P<0.01),而且G-1组与G-2组谷胱甘肽(GSH)含量显著降低(P<0.05),虽然G-1组丙二醛(MDA)无显著性差异(P>0.05),但G-2组丙二醛(MDA)含量显著升高(P<0.05)。研究结果表明,葡萄糖浓度高会影响猪卵母细胞线粒体分布,氧化应激水平升高,成熟效率降低,损害早期胚胎的发育潜能。  相似文献   
7.
花䱻卵母细胞发育的组织学和超微结构观察   总被引:1,自引:0,他引:1  
2015年7月至2017年10月在河南省驻马店宿鸭湖水库采集花?(Hemibarbus maculatus Bleeker)雌鱼样本190尾,体长7.12~32.21 cm,体重10.55~330.22 g,采用组织学和扫描电子显微镜技术观察了花?卵母细胞发育各时期的特征。结果表明,花?卵母细胞发育可分为5个时相,第I时相卵母细胞处于卵原细胞增殖阶段;第Ⅱ时相卵母细胞处于初级生长阶段,出现滤泡膜;第Ⅲ时相卵母细胞出现皮质液泡,细胞质膜之间形成放射带;第Ⅳ时相卵母细胞处于大生长后期,卵黄颗粒增多。电镜下观察发现放射带表面形成微孔状结构,核仁外排,可能与卵母细胞内营养物质积累有关;第Ⅴ时相卵母细胞中细胞核消失,卵母细胞发育为成熟卵子,与卵膜脱离,准备排卵。繁殖季节,花?卵巢成熟系数达到13.78%~17.04%。研究结果可为花?人工繁殖和育种工作提供参考。  相似文献   
8.
This work proposes a computer vision procedure for counting Twospot astyanax (Astyanax bimaculatus) oocytes in Petri dishes using images captured by smartphone. First, the proposed procedure uses simple linear iterative clustering (SLIC) to divide the images into groups of pixels (superpixels). Then, based on their color and space characteristics, the images are classified into light background, dark background, dirt, or oocyte by a machine learning algorithm. Five different types of machine learning algorithms were tested: support vector machines (SVM), decision trees using the algorithm J48 and random forest, k-nearest neighbors (k-NN), and Naive Bayes. To train the algorithms, 8.578 superpixels were classified by an expert into oocyte (n = 354), dirtiness (n = 651), dark background (n = 3.622), and light background (n = 3.951). Of the five learning algorithms, SVM obtained the best result with 97% correct oocyte recognition. Given the wide availability of smartphones, we therefore conclude that the presented procedure can be a valuable tool in future experiments and studies on fertilization and hatching success in Twospot astyanax.  相似文献   
9.
In spite of many attempts to establish an in vitro fertilization (IVF) technique in the equine, no efficient conventional IVF technique is available. The presence of oviductal fluid or oviductal cells during IVF helps to improve embryo production in vitro but is not sufficient to reach high fertilization rates. Thus, our aim was to perform equine IVF either after sperm pre‐incubation with oviductal fluid or in the presence of oviductal cells, and to evaluate the effect of cumulus removal from the oocyte or sperm pre‐incubation with progesterone. In experiments 1 and 2, IVF was performed in the presence of porcine oviduct epithelial cells. The removal of cumulus cells from equine oocytes after in vitro maturation tended to increase the percentage of fertilization when fresh sperm was used (1/33 vs. 4/31, p > 0.05) but had no effect when frozen sperm was used (1/32 vs. 1/32). Equine sperm pre‐incubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/14 vs. 2/18 for fresh, 1/29 vs. 1/25 for frozen). In experiments 3 and 4, IVF was performed after pre‐incubation of sperm with porcine oviductal fluid. The removal of cumulus cells tended to increase the percentage of fertilization when fresh sperm was used (1/24 vs. 3/26, p > 0.05). Sperm pre‐incubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/39 vs. 2/36 for fresh, 2/37 vs. 1/46 for frozen), but two 3–4 cell stage zygotes were obtained with fresh sperm pre‐incubated with progesterone. This is an encouraging result for the setting up of an efficient IVF procedure in equine.  相似文献   
10.
The objectives of present study were to evaluate the effect of casein kinase 1 (CK1) inhibition D4476 on in vitro maturation (IVM) and developmental competence of bovine oocytes. The cumulus oocyte complexes (COCs) were cultured in maturation medium with D4476 (0, 2, 5, 10, 20 μM) for 24 hr. After IVM and in vitro fertilization, through expansion average scores of cumulus cells (CCs), oocyte maturation efficiency, cleavage rate and blastocyst rate of zygote, we found 5 μM D4476 could increase the development potential of oocytes. After the COCs were treated with 5 μM D4476, the results of quantitative real‐time PCR analysis, Lichen red staining and PI staining showed that under without affecting germinal vesicle breakdown and nuclear morphology, D4476 could significantly decrease CK1 and upregulate TCF‐4 in oocytes. Furthermore, without influencing the level of Bad and CTSB, D4476 could significantly increase the expression of β‐catenin, TCF‐4, Cx43, MAPK, PTGS‐2, PTX‐3, TGS‐6, Bax and Bcl‐2 in CCs. Western blot analysis revealed that the addition of 5 μM D4476 during the maturation of COCs resulted in a lower level of Cx43 protein at 12 hr and a higher expression of Cx43 protein at 24 hr compared to the group without D4476. These results indicate that adding optimum D4476 (5 μM) to maturation medium is beneficial to maturity efficiency and development competence of bovine oocytes.  相似文献   
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