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91.
92.
JIANG Xun ZENG Yao-ying HE Xian-hui XU Li-hui DI Jing-fang FENG Zheng ZHAO Jing-xian WANG Qing WANG Tong SHI Jian-bo 《园艺学报》2004,20(6):924-928
AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37% at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage. 相似文献
93.
AIM: To observe effects of homocysteine and antagonized effects of taurine on electronic leakage and free radical production in myocardial mitochondria. METHODS: Myocardial mitochondria of rat heart was isolated, and was broken by supersonic wave to prepare submitochondria. Recombinant of succinic acid cytochrome c reductase was prepared with mitochondria of porcine heart. They were co-incubated with homocysteine and/or taurine with various concentration. The H2O2 and O2- were determined by chemiluminescence methods. The taurine transporter of heart mitochondria and its propert, and effects of homocysteine on its function were studied with glass filter. RESULTS: Homocysteine stimulated oxygen free radical production in heart mitochondria, submitochondria, and succinic acid cytochrome c in a concentration-dependent manner. Although taurine itself did not affect oxygen free radical production, taurine did inhibit oxygen free radical production in mitochondria, submitochondria and succinic acid cytochrome c in a concentration-dependent manner. Taurine transporters of Na+-dependent were existed in mitochondria membrane. Homocysteine inhibited taurine transtport in mitochondria in a concentration-dependent manner. CONCLUSIONS: Taurine inhibited electronic leakage and oxygen free radical production induced by homocysteine in electron transport chain. There were taurine transporters in mitochondria membrane, and transport functions of taurine transporter were inhibited by homocysteine. 相似文献
94.
AIM: To investigate the role of adrenomedullin (AM) in diabetic nephropathy. METHODS: We observed the changes in the expression and secretion of AM, TGF-β1 in the cultured human mesangial cells under high glucose condition and the contents of the laminin and type IV collagen in the supernatants. The effect of intervention with AM was also observed. RESULTS: High glucose condition resulted in increase in the expression and secretion of AM、 TGF-β1、 laminin and type IV collagen. AM reversed the influence of high glucose on the cultured human mesangial cells. CONCLUSION: These results showed that high glucose condition is one of stimulating factors of AM and the renal protective action of AM may be associated with suppression of TGF-β1 and reducing excessive accumulat ion of laminin and type IV collagen. 相似文献
95.
PRRS ELISA试剂盒检测猪繁殖与呼吸综合征病毒抗体的应用 总被引:2,自引:0,他引:2
利用重组杆状病毒表达的PRRSV核衣壳蛋白作抗原制成ELISA诊断试剂盒,检测人工感染PRRSV猪血清、疫苗免疫抗体和田间血清,并与IDEXX公司生产的试剂盒进行比较。结果表明,该试剂盒在PRRSV感染后8天内就可检出感染性抗体,而用IDEXX公司生产的试剂盒在感染后的18天才检测到感染性抗体,对弱毒疫苗的免疫检测也基本能反映出疫苗的免疫应答状况。对华东地区PRRSV流行病学调查结果表明,PRRSV在国内普遍存在,同时也证明研制的试剂盒,是客观科学调查我国PRRS流行情较理想的检测工具。 相似文献
96.
通过 PCR技术 ,以马立克氏病病毒 (MDV) RB1B株基因组 DNA为模板 ,扩增了包括 pp38基因及其启动子 -增强子和终止子在内的 2 2 0 0 bp的核酸片段。将该片段用 Sac 和 Sph 酶定向克隆到 p U C18质粒中 ,构建成重组质粒 ;将重组质粒转染鸡胚成纤维细胞 (CEF) ,用小鼠抗大肠杆菌表达的 pp38血清作间接免疫荧光试验 ,验证 pp38基因的表达。结果 ,在转染的细胞浆中看到了绿色的荧光 ,证实了该启动子的单向启动活性 相似文献
97.
98.
猫细小病毒NS部分基因的克隆及序列分析 总被引:1,自引:0,他引:1
将疫苗用猫细小病毒(FPV)株提取基因组DNA,针对NS特定片段设计引物,利用PCR扩增出了部分基因并将该基因克隆到pMD18-T Vector中测序.结果表明,该基因长613bp,编码204个氨基酸.分离株基因与犬的细小病毒(CPV)、貂的细小病毒(MEV)毒株核苷酸同源性均为99%.氨基酸同源性达到97%、98%以上,与犬、貂的肠炎病毒有密切的亲缘关系. 相似文献
99.
马立克氏病病毒pp38基因启动子和增强子的克隆和序列分析 总被引:4,自引:0,他引:4
参考GeneBank发表的马立克氏病病毒(MDV)国际标准强毒株GA的基因序列,设计合成一对引物,分别以RBIB,814,GD2(广东分离株),J-1-E(北京分离株),Md11,Md5,CV1988等不同毒株的MDV基因组DNA为模板,通过PCR扩增,获得了预期大小的PCR产物。该产物经pGEM-T-easy克隆后测序,将所得序列进行比较分析。结果发现:不同毒株间pp38基因的启动子和增强子序列间有缺失突变,序列的同源性大于95.9%,其中大多数的突变发生在MDV复制的原点附近。 相似文献
100.