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41.
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Smith R Thomas JS 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2002,31(1):26-32
Background — Thrombocytopenia is a common disorder in dogs and development of an objective diagnostic assay to measure platelets newly released from bone marrow into the blood would provide a noninvasive way to predict megakaryocytopoiesis. Reticulated platelets are newly released platelets with increased concentrations of RNA that can be detected by flow cytometric analysis of blood stained with thiazole orange (TO).
Objectives — The goals of this study were to establish a reproducible method to quantitate reticulated platelets in dogs, to establish a reference interval for reticulated platelet percentages in healthy dogs, and to determine whether the percentage of reticulated platelets was nonspecifically increased in nonthrombocytopenic dogs with clinical disease.
Methods — Blood samples were obtained from healthy dogs and from nonthrombocytopenic dogs presented for a variety of disorders. An aliquot of whole blood was stained with TO and a phycoerythrin-labeled monoclonal antibody to platelet CD61, then analyzed by flow cytometry.
Results — The coefficients of variation were 7.8% to 15.6% (intra-assay precision) and 6.1% to 19.5% (interassay precision). Overnight storage for 18 to 26 hours, under variable conditions, resulted in an increase in the percentage of platelets staining with TO. The reference interval for reticulated platelets in the healthy control group was 0–4.3% (0–12,095/μL). No significant differences were found in the mean percentage of reticulated platelets or absolute concentration of reticulated platelets between control and affected dogs.
Conclusions — These studies demonstrate a reliable, noninvasive diagnostic assay for measurement of reticulated platelets in whole blood and provide a baseline for assessment of the clinical utility of the assay. 相似文献
Objectives — The goals of this study were to establish a reproducible method to quantitate reticulated platelets in dogs, to establish a reference interval for reticulated platelet percentages in healthy dogs, and to determine whether the percentage of reticulated platelets was nonspecifically increased in nonthrombocytopenic dogs with clinical disease.
Methods — Blood samples were obtained from healthy dogs and from nonthrombocytopenic dogs presented for a variety of disorders. An aliquot of whole blood was stained with TO and a phycoerythrin-labeled monoclonal antibody to platelet CD61, then analyzed by flow cytometry.
Results — The coefficients of variation were 7.8% to 15.6% (intra-assay precision) and 6.1% to 19.5% (interassay precision). Overnight storage for 18 to 26 hours, under variable conditions, resulted in an increase in the percentage of platelets staining with TO. The reference interval for reticulated platelets in the healthy control group was 0–4.3% (0–12,095/μL). No significant differences were found in the mean percentage of reticulated platelets or absolute concentration of reticulated platelets between control and affected dogs.
Conclusions — These studies demonstrate a reliable, noninvasive diagnostic assay for measurement of reticulated platelets in whole blood and provide a baseline for assessment of the clinical utility of the assay. 相似文献
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Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs 总被引:5,自引:0,他引:5
Erika Varkonyi-Gasic Rongmei Wu Marion Wood Eric F Walton Roger P Hellens 《Plant methods》2007,3(1):12
MicroRNAs (miRNAs) are a class of small non-coding RNAs with a critical role in development and environmental responses. Efficient
and reliable detection of miRNAs is an essential step towards understanding their roles in specific cells and tissues. However,
gel-based assays currently used to detect miRNAs are very limited in terms of throughput, sensitivity and specificity. Here
we provide protocols for detection and quantification of miRNAs by RT-PCR. We describe an end-point and real-time looped RT-PCR
procedure and demonstrate detection of miRNAs from as little as 20 pg of plant tissue total RNA and from total RNA isolated
from as little as 0.1 μl of phloem sap. In addition, we have developed an alternative real-time PCR assay that can further
improve specificity when detecting low abundant miRNAs. Using this assay, we have demonstrated that miRNAs are differentially
expressed in the phloem sap and the surrounding vascular tissue. This method enables fast, sensitive and specific miRNA expression
profiling and is suitable for facilitation of high-throughput detection and quantification of miRNA expression. 相似文献
45.
南方菜豆花叶病毒(SBMV)主要有2株系:菜豆株系(SBMV-B)和豇豆株系(SBMV-C),血清学方法不能予以区分。根据已报道的核酸序列设计了2对特异引物,进行RT-PCR扩增并克隆到载体Bluescript中,序列测定予以证实后利用随机引物法合成放射性cDNA探针,再进行体外转译合成地高辛UTP标记的RNA探针。2种探针均可分别特异地检测Northernblot膜上的SBMV-B和SBMV-C。RNA探针检测SBMV-B和SBMV-C灵敏度分别为:0.1μg和0.01μg。 相似文献
46.
Tropical Animal Health and Production - 相似文献
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Evaluation of existing on-farm agroforestry plots should provide useful supplementary information for the design of improved agroforestry systems, in both research and development projects. Such evaluation has been little used, however, largely due to the methodological difficulties of surveying highly variable on-farm plots, and difficulties in identifying key variables for measurement. This paper describes a set of methods and tools used in evaluating plots of alley-cropping and tree borders around crop fields established by farmers working with the CARE Agroforestry Extension Project in western Kenya. Details of survey design, sampling, and implementation are discussed, and suggestions made for carrying out agroforestry surveys in other projects. A condensed version of the questionnaire is appended. 相似文献
49.
Within-leaf variations in cell size, mitochondrial numbers and dark respiration rates were compared in the most recently expanded tip, the mid-section and base of needles of Pinus radiata D. Don trees grown for 4 years in open-top chambers at ambient (36 Pa) or elevated (65 Pa) carbon dioxide partial pressure (p(CO2)a). Mitochondrial numbers and respiratory activity varied along the length of the needle, with the highest number of mitochondria per unit cytoplasm and the highest rate of respiration per unit leaf area at the base of the needle. Regardless of the location of the cells (tip, middle or basal sections), needles collected from trees grown in elevated p(CO2)a had nearly twice the number of mitochondria per unit cytoplasm as those grown in ambient p(CO2)a. This stimulation of mitochondrial density by growth at elevated p(CO2)a was greater at the tip of the needle (2.7 times more mitochondria than in needles grown in ambient CO2) than at the base of the needle (1.7 times). The mean size of individual mitochondria was unaffected either by growth at elevated p(CO2)a or by position along the needle. Tree growth at elevated p(CO2)a had a variable effect on respiration per unit leaf area, significantly increasing respiration in the tip of the needles (+25%) and decreasing respiration at the mid-section and base of the needles (-14% and -25%, respectively). Although a simple relationship between respiration per unit leaf area and mitochondrial number per unit cytoplasm was found within each CO2 treatment, the variable effect of growth at elevated p(CO2)a on respiration along the length of the needles indicates that a more complex relationship must determine the association between structure and function in these needles. 相似文献
50.
Stem cuttings of Eucalyptus globulus are used within tree improvement programs and for mass deployment. To be successful, cuttings must perform as well or better than seedlings. The root systems of cuttings are fundamentally different from those of seedlings. If these differences influence growth, the differences and their consequences must be identified and the propagation system manipulated to improve performance of the propagules.Cuttings are only a viable alternative to seedlings as planting stock if the method of propagation does not affect their growth and development adversely. Full-sibling cuttings and seedlings of Eucalyptus globulus were compared under controlled environmental conditions to minimise extraneous sources of variation, and to establish whether changes in growth or development were induced by propagation. On three occasions over a period of eight weeks root-collar diameter, shoot height, leaf and stem weight, shoot/root ratios and root system morphology were measured on cuttings and seedlings. Seedlings were taller than cuttings throughout the experiment, but both plant types had similar height growth rates. Diameter growth rates were lower in cuttings than seedlings, and there were differences in both height and diameter growth rates between families. Root system configuration differed between the plant types. Seedlings had strongly gravitropic tap-roots, with two types of primary roots from which secondary roots emerged. Cuttings had no tap roots, and the main structural components of their root systems were adventitious roots formed during propagation. Cuttings did not develop further structural roots during the experiment, whereas seedlings continued to develop primary roots. Individual primary roots of cuttings were longer and had larger mid-point diameters than those of seedlings, but the total length of primary roots was greater in seedlings. Seedlings also had a greater number and total length of secondary roots. Shoot/root ratios, calculated from a range of functional measures, were higher in cuttings than seedlings. 相似文献