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以卷丹(Lilium lancifolium)叶腋组织为研究材料,利用RT-PCR和RACE技术克隆得到AGO1基因的cDNA全长,命名为LlAGO1。其全长4 014 bp,开放阅读框长3 687 bp,编码1 228个氨基酸残基,其编码蛋白分子量为135.36 kD,理论等电点(pI)为9.57。氨基酸序列分析表明LlAGO1含有PAZ和Piwi两个AGO1典型的结构域;信号肽预测结果表明LlAGO1蛋白不存在信号肽,为非分泌蛋白;亚细胞定位预测其主要定位于细胞核;与相关同源蛋白高度相似,且与芦笋AGO1a蛋白(XP_020260210.1)亲缘关系最近。qRT-PCR分析结果表明:LlAGO1在卷丹叶腋、鳞片、根、叶等不同组织均有表达,其中在叶腋中的表达量最高,叶片和根中的表达较弱;腋生珠芽形成过程中,LlAGO1仅在可形成珠芽的上部叶腋表达,且在珠芽形成时表达量最高,而在不形成珠芽的下部叶腋几乎不表达,推测LlAGO1可能与卷丹珠芽的形成相关。 相似文献
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DAI Pei GAO Fen GAO Hong-wei WANG Yuan FENG Gao-jie ZHANG Qin-feng BAI Rui QIN Wei-wei LI Hong SONG Xiao-su 《园艺学报》2019,35(2):212-217
AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells. 相似文献
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AIM:To observe changes of serum asymmetry dimethylarginine (ADMA) and endothelin-1 (ET-1) levels,which reflect blood vessel endothelial function,after therapy of enhanced external counterpulsation(EECP) in patients who had taken percutaneous transluminal coronary angioplasty and stent.METHODS:Fifty one coronary heart disease patients (all of them had taken percutaneous tranluminal coronary angioplasty and stent) were distributed into two groups by matching them with ratio of 1∶2,17 patients in EECP group and 34 patients in control group.Both of two groups were given conventional medicine,in addition,EECP group was undertaken three courses of treatment of EECP.ADMA was detected by HPLC-fluorescence method,and ET-1 was detected by radio-immunity method.RESULTS:In EECP group ,compared with prior treatment,ADMA and ET-1 levels was obviously reduced (P<0.05).In control group,compared with prior treatment,ADMA had no significant difference (P>0.05) and ET-1 was higher (P<0.01).Compared between two groups,extent of decrease in ADMA and ET-1 levels in EECP group were more obvious than control group (P<0.01).Improvement of angina and decrease of frequency of angina was more obvious in EECP group than control group.Besides,decrease of ADMA level was positive correlation with them (r=0.85,0.70,respectively P<0.01).CONCLUSION:EECP reduces serum ADMA and ET-1 levels by increasing shear stress to vessel endothelium in coronary heart disease patients who had taken percutaneous tranluminal coronary angioplasty.It hints that EECP can improve endothelial function and provides experimental evidence for the combined treatment of coronary heart disease. 相似文献
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全蚀病是小麦上一种重要的土传病害。选育和种植抗病品种是防治小麦全蚀病的根本途径,抗病基因研究是抗病育种的基础性工作。根据基因TaWIR1b(Accession no.M94959.1)的全长序列设计引物扩增‘新农19’的cDNA,获得了完整ORF,编码85个氨基酸残基,比对后发现与TaWIR1b序列同源性达100%。根据获得的TaWIR1b基因全长序列设计定量引物,分析TaWIR1b在全蚀菌胁迫条件下不同互作模式的表达特征。结果表明接种全蚀病菌后抗病小麦品种‘新农19’中TaWIR1b基因被诱导表达,接菌后3d达到峰值143.97,感病品种‘新麦19’中峰值出现在接菌后8d,表达量仅为对照的4.22倍,提示该基因可能参与小麦对全蚀病的抗病过程。 相似文献