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为探讨不同检测材料对ELISA检测禽白血病病毒(ALV)-p27抗原结果的影响,试验首先对180日龄地方品种母鸡的泄殖腔棉拭子ALV-p27抗原进行ELISA检测,选取部分阳性鸡和阴性鸡进行分组试验,采用ELISA对试验鸡血清、蛋清和病毒分离的细胞培养液进行ALV-p27抗原检测,采用RT-PCR方法检测血液ALV特异性基因。结果显示,阳性组中以血清、蛋清和细胞培养液作为ELISA ALV-p27抗原检测材料,其检测阳性率均低于泄殖腔棉拭子,血清检测的阳性个体包含全部蛋清和细胞培养液ELISA检测的阳性个体。ELISA检测数据的相关性分析显示,只有血清和细胞培养液检测数据间存在显著性相关,线性关系方程为Y(细胞上清)=1.8439X(血清)-0.1469,R2=0.937;阳性组中ALV-p27基因检测阳性率低于泄殖腔棉拭子,但高于血清、蛋清和细胞培养液,其包含所有血清ELISA检测的阳性样品;外源性ALV-J gp85基因阳性率仅为29.17%,且阳性样品均属于血清ELISA阳性样品。综上所述,成年鸡以泄殖腔棉拭子作为ELISA ALV-p27抗原检测材料存在假阳性结果,蛋清和细胞培养液作为检测材料存在漏检的可能,血清作为ELISA检测材能够更准确地反映成年鸡群ALV感染状态。 相似文献
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森林草莓独脚金内酯合成关键基因D27的克隆与表达分析 总被引:2,自引:0,他引:2
根据森林草莓中独角金内酯合成过程中的关键基因D27的序列信息设计引物,获得了D27基因的完整开放阅读框架。采用MEGA5.0软件,对森林草莓与其他物种D27基因编码的氨基酸序列进行聚类分析,发现森林草莓D27基因与其他物种D27基因具有较高的同源性。设置正常供磷、缺磷和在缺磷条件下接种丛枝状菌根真菌处理,利用电感耦合等离子发射光谱仪测定草莓植株的磷含量,结果表明缺磷条件下接种丛枝状菌根真菌,草莓植株中磷含量极显著提高,而缺磷处理的磷含量极显著降低。实时荧光定量PCR结果表明,与正常供磷处理相比,缺磷和接种丛枝状菌根真菌处理条件下,D27基因的表达量分别上调7倍和11倍,这说明D27基因的表达既受到磷水平调控,又受到菌根形成的调控。 相似文献
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AIM:To investigate the effect of cholesterol metabolite 27-hydroxycholesterol (27-OHC) on the proliferation of lung cancer cells. METHODS:Human lung cancer A549 cells were treated with 27-OHC at different concentrations (0, 0.3125, 0.625, 1.25, 2.5, 5 and 10 μmol/L) for 24~48 h. The cell viability, cell cycle, cell prolife-ration, the intracellular cholesterol levels and cholesterol metabolism-related molecule expression were subsequently assessed by CCK-8 assay, flow cytometry, EdU staining, tissue total cholesterol detection kit, real-time PCR and Western blot. RESULTS:27-OHC decreased the viability of the A549 cells in a dose-and time-dependent manner (P<0.01) and inhibited the cell proliferation (P<0.05). The expression of typical liver X receptor (LXR) downstream target proteins including ATP-binding cassette transporter A1 (ABCA1), low-density lipoprotein receptor (LDLR), and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CR) were modulated, which promoted the efflux of intracellular cholesterol, and reduced cholesterol influx and de novo synthesis, resulting in decreased intracellular cholesterol levels and cell viability. Furthermore, the inhibitory effect of 27-OHC on A549 cell viability was significantly attenuated after the LXR pathway was partially blocked by 5 μmol/L GSK2033 treatment (P<0.05). CONCLUSION:27-OHC inhibits A549 cell prolife-ration via activation of LXR signaling pathway. 相似文献
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LIU Lian YU Rong-jie DAI Yun ZENG Zhi-xing GUO Xiao-ling JI Qing-shan ZHONG Jing-xiang 《园艺学报》2015,31(1):135-140
AIM: To construct the prokaryotic expression system containing protein transduction domain (PTD) with heat shock protein 27 (HSP27) in order to prepare and purify the recombinant protein, and to verify whether the recombinant protein PTD-HSP27 has the ability to penetrate the human lens epithelial cell (HLEC) membrane and the rabbit cornea. METHODS: The plasmid pKYB-PTD-HSPB1-6His was constructed by the technique of overlap extension PCR. The plasmid was transformed and PTD-HSP27 was purified through nickel affinity chromatography column and identified by Western blotting. PTD-HSP27-6His was labeled with the fluorescein isothiocyanate (FITC). The penetrating ability of PTD-HSP27 into HLECs and rabbit cornea was tested. RESULTS: The recombinant PTD-HSP27 plasmid was successfully cloned and effectively expressed. The correctness of the recombinant protein PTD-HSP27 was demonstrated. Fluorescence microscopic examination showed that PTD-HSP27-FITC was internalized by HLECs. Fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor. CONCLUSION: The recombined gene PTD-HSPB1 was constructed by overlap extension PCR technique and the PTD-HSP27 fusion protein was prepared and purified by nickel affinity chromatography column. Using the technique of PTD-fusion protein, HSP27 was transduced into HLECs and passed through the cornea. 相似文献
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目的:探讨HSP27、p14^ARF和caspase-3蛋白在大肠癌中的表达及其与大肠癌I临床病理特征的关系。方法:利用sP法检测大肠癌48例和大肠腺瘤10例以及正常大肠黏膜组织15例中HSP27、p14^ARF和caspase-3蛋白的表达。结果:大肠癌组织中HSP27、p14^ARF和caspase-3蛋白阳性表达率分别为79.2%、58.4%和66.7%,大肠腺瘤组织中分别为40.0%、70.0%和100.0%,正常大肠黏膜组织中分别为33.3%、80.0%和93.3%。p14^ARF和caspase-3蛋白的表达在高、中分化癌组的表达率分别为64.1%、74.4%,明显高于低分化组22.2%、33.3%(Pd0.05)。结论:HSP27、p14^ARF和caspase-3与大肠癌的发生有关,可作为大肠癌临床评价肿瘤生物学行为的指标。 相似文献
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