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Sorting of membrane and fluid at the apical pole of polarized Madin-Darby canine kidney cells 下载免费PDF全文
When fluid-phase markers are internalized from opposite poles of polarized Madin-Darby canine kidney cells, they accumulate in distinct apical and basolateral early endosomes before meeting in late endosomes. Recent evidence suggests that significant mixing of apically and basolaterally internalized membrane proteins occurs in specialized apical endosomal compartments, including the common recycling endosome and the apical recycling endosome (ARE). The relationship between these latter compartments and the fluid-labeled apical early endosome is unknown at present. We report that when the apical recycling marker, membrane-bound immunoglobulin A (a ligand for the polymeric immunoglobulin receptor), and fluid-phase dextran are cointernalized from the apical poles of Madin-Darby canine kidney cells, they enter a shared apical early endosome (=2.5 min at 37 degrees C) and are then rapidly segregated from one another. The dextran remains in the large supranuclear EEA1-positive early endosomes while recycling polymeric immunoglobulin receptor-bound immunoglobulin A is delivered to a Rab11-positive subapical recycling compartment. This latter step requires an intact microtubule cytoskeleton. Receptor-bound transferrin, a marker of the basolateral recycling pathway, has limited access to the fluid-rich apical early endosome but is excluded from the subapical elements of the Rab11-positive recycling compartment. We propose that the term ARE be used to describe the subapical Rab11-positive compartment and that the ARE is distinct from both the transferrin-rich common recycling endosome and the fluid-rich apical early endosome. 相似文献
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Midgley RS Blake NW Yao QY Croom-Carter D Cheung ST Leung SF Chan AT Johnson PJ Huang D Rickinson AB Lee SP 《Journal of virology》2000,74(3):1544-1548
Among 34 Epstein-Barr virus isolates from nonimmunocompromised Chinese donors, we identified three intertypic recombinants with type 1 sequences at the EBNA2 locus and type 2 sequences at some or all of the EBNA3A, -3B, and -3C loci. These appear to have arisen from independent, evolutionarily recent recombination events; such events may be commoner in nonimmunocompromised populations than hitherto imagined. 相似文献
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Christina F. Leung Sarah E. Webb rew L. Miller 《Development, growth & differentiation》1998,40(3):313-326
Through the injection of f -aequorin (a calcium-specific luminescent reporter), and the use of an imaging photon detector, transient localized elevations of free cytosolic calcium in the forming blastodisc (BD) and animal hemisphere cortex were visualized that correlated with ooplasmic segregation. The introduction of an appropriate concentration of the weak (KD = 1.5 μmol/L) calcium buffer 5,5'-dibromo-BAPTA results in the dissipation of these calcium domains, and inhibits cytoplasmic streaming and the subsequent formation of a BD at the animal pole. These inhibitory actions are dependent on the final cytosolic concentration of buffer within the egg: ≥ 1.3 mmol/L blocks ooplasmic streaming; < 1.3 mmol/L eggs segregate normally. Injection of 5,5'-dimethyl-BAPTA (KD = 0.15 μmol/L) to a final concentration of 1.5 mmol/L as a control has no effect on ooplasmic streaming. These results suggest that localized domains of elevated free cytosolic calcium are essential for ooplasmic segregation in zebrafish. Furthermore, a hypothetical model is presented linking these calcium transients to the contraction of a cortically located actin microfilament network as a possible mechanism providing the driving force for segregation. 相似文献
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Evidence for the molecular self-assembly of amelogenin proteins to form quasi-spherical particles (“nanospheres”) in solution, bothin vitroandin vivo,has recently been documented. A particle-size distribution analysis of dynamic light-scattering data was undertaken to investigate the influence of temperature on this molecular self-assembly process at three different pH's. The long-term objective was to correlate these observations to the unusual physiochemical characteristics of the protein, to improve understanding of the molecular mechanisms involved in the generation of amelogenin “nanospheres” and understanding of their putative relation to amelogenin functionin vivo. We analyzed data using two different algorithms: Dynamics and DynaLS. It was found that at pH 8, in a temperature range between 5 and 25°C, the size of the recombinant amelogenin nanospheres is monodisperse, giving rise to particles of 15–18 nm in hydrodynamic radius. However, heterogeneous distribution of particle size was observed at temperature ranges between 27 and 35°C, becoming monodisperse again with larger particles (60–70 nm) after the temperature was elevated to 37–40°C. We interpret these results to suggest that amelogenin molecular self-association possesses a second stage assembly process at temperatures of 30–35°C, creating larger entities which apparently are structured and stable at 37–40°C. The effect of pH on the size of amelogenin “aggregates” was much more noticeable at 37°C compared to that at 25°C. This observation suggests that at physiological temperature (i.e., 37°C) amelogenin molecular self-assembly is extremely sensitive to pH changes. This finding supports the notion that local pH changes in the microenvironment of the enamel extracellular matrix may play critical roles in controlling the structural organization of the organic matrix framework. 相似文献
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附睾内液体微环境对精子的成熟和贮藏是相当重要的。附睾液体的形成取决于附睾上皮的吸收与分泌功能,而先前的实验已经证明:这些吸收与分泌的活动是受到除神经激素以外旁分泌与自分泌的调控。虽然肾素血管紧张素系统(RAS)在很多组织上旁分泌与自分泌的作用已多有报导,但其在附睾的存在及作用仍鲜为人知。本综述总结了本实验室在这方面的研究结果,通过使用不同的实验方法,例如,免疫组织化学,放射免疫分析,分子生物学及短路电流电生理学方法,我们得到的结果显示了RAS主要成员在附睾的分布(Fig.1)和表达,并阐明了血管紧张素II对附睾阴离子分泌的调控作用(Figs.2&3)。本综述还就血管紧张素II对附睾旁分泌与自分泌的作用及其机制(Fig.4),以及对精子功能的作用进行了讨论。 相似文献