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AIM: This study was designed to investigate the secretion of VEGF and its receptor (flt-1 or flk-1/KDR) protein by cultured bovine thoracic aortic endothelial cells treated with various insulin concentrations. METHODS: Endothelial cells was isolated from bovine thoracic aorta, and cultured in serum-free medium, then incubated with different insulin concentrations (30 mU/L, 300 mU/L, 3 000 mU/L). The level of VEGF and its receptor (flt-1 or flk-1/KDR) protein were detected by immunohistochemical staining. RESULTS: As compared with no insulin group, the expression of VEGF protein in low insulin concentration (30 mU/L and 300 mU/L) groups were significantly increased (P<0.01). The expression of VEGF protein in high insulin concentration (3 000 mU/L) group was significantly decreased (P<0.05). Howerer, no difference of the expression of VEGF receptor (flt-1 or flk-1/KDR) protein among all groups (P>0.05) was observed. CONCLUSION: Low concentration insulin up-regulates the VEGF protein expression while high concentration insulin down-regulates the VEGF protein expression in bovine thoracic aortic endothelial cells, but insulin had no directly effect on the VEGF receptor (flt-1 or flk-1/KDR) protein expression in bovine thoracic aortic endothelial cells. 相似文献
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LUO Ming-hao HU Shu-peng WANG Rui-yu Li Chang Lü Ding-yi YANG Xi-yang LUO Su-xin 《园艺学报》2000,36(10):1739-1744
AIM To investigate whether interleukin-1β (IL-1β) regulates endothelial nitric oxide synthase (eNOS) phosphorylation at Ser1177 site in human umbilical vein endothelial cells (HUVECs), and to explore its possible mechanism. METHODS The HUVECs were randomly divided into normal control group, tumor necrosis factor-α (TNF-α) group, IL-1β group, IL-6 group, SC79 [protein kinase B (PKB/AKT) specific agonist] group and SC79+IL-1β group. Western blot was used to determine the protein levels of eNOS, p-eNOS-Ser1177, AKT and p-AKT-Ser473 in the HUVECs. Chemical colorimetry was used to detect the nitric oxide (NO) content in the culture medium of HUVECs. RESULTS No statistically significant difference of p-eNOS-Ser1177 level in HUVECs treated with TNF-α and IL-6 was observed as compared with normal control group (P >0.05), while the protein level of p-eNOS-Ser1177 in the HUVECs and the content of NO in the culture medium of HUVECs decreased significantly in IL-1β group (P <0.05), and the protein level of p-AKT-Ser473 in the HUVECs was decreased as compared with normal control group (P <0.05). The AKT agonist SC79 blocked the down-regulation effect of IL-1β on p-eNOS-Ser1177 level in the HUVECs and NO content in the culture medium of HUVECs (P< 0.05). CONCLUSION IL-1β down-regulates the protein level of p-eNOS-Ser1177 in HUVECs and affects the activity of eNOS, which may be involved in AKT/eNOS signaling pathway. 相似文献
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[目的]建立稳定快速的慢病毒转基因羊DNA水平检测方法。[方法]采集初生转基因羔羊的子叶、脐带、尾组织以及死亡羔羊的全身各组织,提取基因组DNA作为模板进行PCR检测,以Follistatin基因N+D1片段特异性引物进行PCR检测;同时,对慢病毒载体的CMV启动子、5’-LTR等结构元件进行PCR检测;提取死亡羔羊的全身组织以及转基因羔羊的活体肌肉组织总RNA,进行RTPCR检测。[结果]采用试剂盒提取的基因组DNA比常规方法提取的DNA快速、高效,且更易被检测,扩增时的引物采用载体上游与目标基因下游的组合大大增加了检测的特异性,避免了内源性目标基因的干扰而造成假阳性;初生转基因羔羊的尾组织可作为PCR检测的组织样品,其结果可靠、准确;对CMV启动子及5’-LTR等结构元件作PCR检测为转基因动物的安全性考察提供了数据,同时验证了转基因羔羊目的基因的检测结果;对转基因羔羊肌肉组织提取总RNA后进行RT-PCR检测,其中有3只羔羊没有检测到转录产物,其他均有转录。[结论]该研究建立的转基因绵羊PCR检测方法高效、快速且准确,为进一步蛋白水平检测提供试验依据,也为建立转基因绵羊系统性多层次的检测方法奠定基础,还为转基因绵羊的安全监测提供了稳定的技术平台。 相似文献
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新疆野苹果F_1代果实性状的相关分析和主成分分析 总被引:1,自引:0,他引:1
以4年生新疆野苹果×富士杂交F1代群体为试材,对19个果实品质指标进行测定,利用相关分析和主成分分析从功能基因研究和品系选育两个角度对各指标的遗传变异进行考察。结果表明:(1)与栽培苹果相比,F1代在果实质地、花色苷有关性状之间的相关性存在差异:钙与果实质地的3个指标相关性不显著;花色苷与糖含量相关性不显著,但与钙、钾含量相关性极显著。钙与多个指标极显著相关,可作为功能基因研究的重点。(2)从主成分贡献率看,果实质地、矿质元素、果实大小、花色苷含量4个主成分在遗传信息中占的比重较大,是下一步功能基因研究的切入点,也是进行品质综合评价的重点考察对象。加大对果实质地、果实大小、花色苷含量的考察可提高育种效率。(3)对主成分综合得分排序,根据不同育种目标选择综合性状优良的个体,是新疆野苹果在育种中利用保存的有效捷径。 相似文献
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MIAO Jin-feng ZHANG Yuan-shu HUANG Guo-qing MA Hai-tian ZOU Si-xiang ZHU Yu-min 《中国农业科学(英文版)》2009,8(8)
The aim of this study was to evaluate, in rats, the changes in the T helper type 1 (Th 1)/Th2 radio in mammary glands after an intramammary infusion of lipopolysaccharide (LPS) and to characterize the moderating effects of the polysaecharide nucleic acid of Bacillus Calraette Guerin (BCG-PSN) on the mammary gland. In the control group, the levels of IL-2 and INF-γ, mRNA expression increased, whereas IL-4 mRNA expression decreased after LPS challenge. As a consequence, the INF-γ/IL-4 mRNA ratio was significantly higher at 3, 6, and 9 h post-infusion (PI) compared to the control value (O h; P<0.01).BCG-PSN increased mRNA expression of both INF-γ' and IL-4 before infusion of LPS. LPS challenge significantly the reduced Th1/Th2 eytokine ratio due to Th1 cytokine IFN-γ suppression and Th2 cytokine IL-4 upregulation compared with the control group. A significant reduction ofN-acety1-β-D-glucosaminidase (NAGase) was observed at 24 h PI in the BCG-PSN treatment group compared to the control group (P<0.05). Thus, it was demonstrated that level of BCG-PSN might change the Th1/Th2 ratio mainly by enhancing the Th2 immune response. This is the first report of a Th1/Th2 change induced by coliform mastitis and characterization of the effect of BCG-PSN on mammary gland inflammation. This study makes a better understanding of the mechanisms of coliform mastitis and provides a putative novel strategy for the prevention and/or treatment of mastitis. 相似文献
20.
为构建牛瑟氏泰勒虫双拷贝p23表面蛋白基因真核表达质粒,根据GenBank牛瑟氏泰勒虫p23表面蛋白基因序列(D84447),分别设计2对特异性引物,利用全血基因组DNA提取试剂盒提取牛瑟氏泰勒虫基因组DNA,采用SOE—PCR技术构建双拷贝p23基因,克隆到pMD-18-T载体上,经过PCR、酶切鉴定及测序后,亚克隆到pVAX-Ⅰ真核表达载体上,经过鉴定后采用脂质体法将重组质粒pVAXI-2p23转染到BHK-21细胞,用IFA和RT—PCR来鉴定目的基因的表达情况.结果表明,成功构建了牛瑟氏泰勒虫双拷贝p23表面蛋白基因真核表达质粒,并在BHK-21细胞中获得表达. 相似文献