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细胞松弛素B浓度对小鼠卵母细胞去核效果的影响 总被引:7,自引:0,他引:7
目的 探讨细胞松弛素 B浓度对小鼠卵母细胞去核效果的影响。方法 常规超排 KM小鼠获卵母细胞 ,将有明显第一极体的卵母细胞放入含有 5× 10 - 3、1× 10 - 2 、2× 10 - 2 g/ L CB的成熟培养液中孵育 15 min,采用盲吸法去核 ,去核后的卵母细胞放入含 5× 10 - 3g/ L Hoechst33342的成熟培养液中染色 15 min,在荧光显微镜下检查去核的效率。结果 CB组去核率可达 88.2 % ,而对照组去核成功率仅达 2 9.7% ,两者的去核效果统计学上有显著差异 ;同时 CB的浓度过高会影响去核的效果 ,使卵母细胞的破溃率升高。结论 CB有利于去核 ,同时卵胞膜可以借助磷脂双层的游动性而很快得到修复 ;高浓度的 CB对卵胞膜作用过于强烈 ,细胞骨架严重破坏 ,磷脂双层结构的弹性和粘性减弱 ,修复能力降低 ,影响胚胎的构建。 相似文献
3.
The present study examines the contribution of the nucleus to meiotic competence in mouse oocytes that were reconstructed using nuclear transfer. Three types of reconstructed oocytes were produced: MP‐GV, by transplanting the male pronucleus (MP) into germinal vesicle (GV) stage oocytes; 3T3‐GV, by transplanting the nucleus of a National Institute of Health (NIH) 3T3 cell into a GV stage oocyte; and 3T3‐MII, by transplanting the nucleus of an NIH 3T3 cell into a metaphase II (MII) stage oocyte. The fusion rates differed, but not significantly, in the MP‐GV, 3T3‐GV, and 3T3‐MII groups (77, 63, 56%, respectively). Then, meiotic competence was compared in MP‐GV, 3T3‐GV and non‐manipulated GV stage oocytes as a control. Nuclear envelope breakdown occurred in all the reconstructed oocytes, as well as the control ones. The percentage of first polar body extrusion differed between the MP‐GV (100%), 3T3‐GV (72%), and control (67%) groups. DNA staining with Hoechst 33342 revealed that in the MP‐GV‐group oocytes that had reached MII stage, the chromosomes were condensed and aligned in a regular array similar to the normal metaphase plate. By contrast, in 3T3‐GV group oocytes, the condensed chromosomes were irregularly scattered in the cytoplasm. These results suggest that the donor nucleus affects meiotic competence in reconstructed oocytes. 相似文献
4.
Effect of dasatinib in a xenograft mouse model of canine histiocytic sarcoma and in vitro expression status of its potential target EPHA2 下载免费PDF全文
K. Ito R. Miyamoto H. Tani S. Kurita M. Kobayashi K. Tamura M. Bonkobara 《Journal of veterinary pharmacology and therapeutics》2018,41(1):e45-e48
Canine histiocytic sarcoma (HS) is an aggressive and highly metastatic tumor. Previously, the kinase inhibitor dasatinib was shown to have potent growth inhibitory activity against HS cells in vitro, possibly via targeting the EPHA2 receptor. Here, the in vivo effect of dasatinib in HS cells was investigated using a xenograft mouse model. Moreover, the expression status of EPHA2 was examined in six HS cell lines, ranging from insensitive to highly sensitive to dasatinib. In the HS xenograft mouse model, dasatinib significantly suppressed tumor growth, as illustrated by a decrease in mitotic and Ki67 indices and an increase in apoptotic index in tumor tissues. On Western blot analysis, EPHA2 was only weakly detected in all HS cell lines, regardless of sensitivity to dasatinib. Dasatinib likely results in the inhibition of xenograft tumor growth via a mechanism other than targeting EPHA2. The findings of this study suggest that dasatinib is a targeted therapy drug worthy of further exploration for the treatment of canine HS. 相似文献
5.
Kyohei Yasuno Masako Imaoka Tetsuya Ohsawa Keiko Okado Kiyonori Kai Yoshimi Tsuchiya 《Journal of toxicologic pathology》2022,35(4):345
Inflammation of the cardiac coronary artery in ICR mice is occasionally observed in toxicity studies; however, this has not been well explored histologically. Herein, we investigated the detailed histology of the associated lesions in 6–8-week-old ICR mice. Coronary artery inflammation in the right ventricular wall was observed in 10 of 142 mice (7.0%). Histopathological examination revealed hypertrophy of the vascular smooth muscle cells and perivascular infiltration of macrophages in mild cases. In moderate to marked cases, single-cell necrosis of vascular smooth muscle cells, hemorrhage of the tunica media, and fibrinoid necrosis of the vessel wall were observed, in addition to the changes seen in mild cases. Electron microscopic examination of moderate cases revealed a discontinuous internal elastic lamina suggestive of rupture, and vascular smooth muscle cells beneath the elastic lamina showed degeneration and necrosis. These findings suggest that the lesions developed as a rupture of the internal elastic lamina and necrosis of vascular smooth muscle cells, while leaked plasma components caused vascular and perivascular inflammation. In ICR mice, dystrophic calcinosis (DCC) is known to occur rarely in the right ventricle. DCC is defined as focal calcification in necrotic myocardial fibers, the pathogenesis of which is considered to involve ectopic calcification. Since calcification was not observed in any part of the heart, including the inflammation region, the pathophysiology of cardiac arterial inflammation seen in our ICR mice was considered to differ from that of DCC. 相似文献
6.
[目的]研究Rho激酶特异抑制剂Y-27632对小鼠早期胚胎发育的影响。[方法]取小鼠早期胚胎2细-胞,经体外培养1~2 d后,进行细胞计数、受体移植,观测Rho激酶抑制剂Y-27632对细胞数目和胚胎着床效率的影响。[结果]早期胚胎经Y-27632处理后,胚胎发育延迟,囊胚出腔脱带时间推后,试验组胚胎桑椹胚、囊胚发育率同对照组差异不显著(P〉0.05),与同期胚胎相比细胞数目增加,差异显著(P〈0.05),着床效率相当。[结论]Rho激酶特异抑制剂Y-27632对小鼠早期胚胎细胞数目增加和延迟着床具有重要作用。 相似文献
7.
Chi-Heung Cho Chang-Jun Lee Min-Gyeong Kim Bomi Ryu Jun-Geon Je Yoonsook Kim Sang-Hoon Lee 《Marine drugs》2022,20(6)
Advanced glycation end-products (AGEs) play a vital role in the pathogenesis of diabetic complications. Methylglyoxal (MGO), one of the major precursors of AGEs, is a highly reactive dicarbonyl compound that plays an important role in the pathogenesis of diabetic nephropathy. This study was designed to evaluate the therapeutic potential of phlorotannin-rich Ecklonia cava extract (ECE) on MGO-induced diabetic nephropathy in in vitro models using mouse glomerular mesangial cells. ECE showed anti-glycation activity via breaking of AGEs-collagen cross-links and inhibition of AGEs formation and AGE-collagen cross-linking formation. The renoprotective effects were determined by assessing intracellular reactive oxygen species (ROS) and MGO accumulation, cell apoptosis, and the Nrf-2/ARE signaling pathway. MGO-induced renal damage, intracellular ROS production level, and MGO-protein adduct accumulation were significantly decreased by pretreating ECE. Moreover, ECE pretreatment exhibited preventive properties against MGO-induced dicarbonyl stress via activation of the Nrf2/ARE signaling pathway and reduction of RAGE protein expression in mouse glomerular mesangial cells. Collectively, these results indicated potential anti-glycation properties and prominent preventive effects of ECE against MGO-induced renal damage. Additionally, ECE may be utilized for the management of AGE-related diabetic nephropathy. 相似文献
8.
BPA不影响卵母细胞减数分裂相关基因Dazl的甲基化 总被引:1,自引:0,他引:1
为了探讨环境雌激素BPA对小鼠卵母细胞减数分裂相关基因Dazl甲基化的影响,本研究通过给孕鼠饮用含有BPA的水方式使胎鼠在发育过程中接触BPA,利用重亚硫酸盐测序法,分析了胎鼠生殖嵴卵母细胞不同发育时期Dazl甲基化水平的变化。结果显示:Dazl在减数分裂期间处于低甲基化水平,无论对照组或处理组均低于10%,说明Dazl的低甲基化对维持减数分裂的正常进行有重要作用;对照组与处理组的甲基化水平相当,差异不显著,说明本研究的BPA浓度不影响卵母细胞Dazl的甲基化水平。 相似文献
9.
The assay was aimed to study the effect of Hedyotis diffusa Willd superfines powder on immune function. Three groups of mice were gavaged with the drugs at doses of 1.75,1.25 and 0.75 g/kg once a day for 7 consecutive days, repectively. The mice of control group were administered with the same volume of saline.At the last time, mice were injected into the muscle with Newcastle disease vaccine,and get peripheral blood on days 7, 14 and 21 after the medicine administration. The enhancement of innate immune responses were evaluated by using CCK-8 method, hemagglutination inhibition test, macrophage phagocytic function test and organ coefficient method. The levels of lymphocytes proliferative capacity, serum antibody, peritoneal macrophage phagocytosis and the indexes of immune organs in mice were extremely significantly enhanced (P<0.01). Hedyotis diffusa dWilld superfines powder could extremely significantly regulate the immunity function of mice (P<0.01). 相似文献
10.
Zhang Wen-yu Xu Jia-hui Zhang Chun-yu Tong Hui-li Li Shu-feng Yan Yun-qin 《东北农业大学学报(英文版)》2021,28(3):38-47
Myoblast differentiation is an essential process during skeletal muscle development. C2 C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro. Dehydrogenase/reductase(SDR family) member 3(DHRS3) is a highly conserved member in short-chain alcohol dehydrogenase/reductase superfamily and has been shown to be involved in the metabolism of retinol. Previous experimental results showed that the expression of DHRS3 increased significantly during the differentiation of myoblasts differentiation. However, the effect of DHRS3 on mouse muscle cell differentiation was unclear. The objective of current study was to determine if DHRS3 affected muscle cell differentiation, and if DHRS3 was involved in muscle regeneration. Protein expression was determined by western blot and immunofluorescence analysis. The activation and inhibition of DHRS3 increased and decreased C2 C12 myoblast differentiation respectively, which indicated that DHRS3 could affect C2 C12 myoblast differentiation. DHRS3 expression was significantly changed during muscle regeneration, with the regeneration of muscle injury, the expression of DHRS3 tended to increase first and then decrease. It suggested that DHRS3 might be involved in muscle regeneration. In summary, this study confirmed the involvement of DHRS3 in C2 C12 myoblast differentiation and mouse skeletal muscle regeneration and provided a theoretical basis for further elucidating the molecular mechanism of muscle development. 相似文献