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大鼠皮质神经元的体外培养和纯化 总被引:8,自引:2,他引:6
目的:探讨大鼠皮质神经元分离纯化的有效方法。方法:取胎鼠大脑皮质细胞,分别在不同的培养液中进行原代培养,利用相差显微镜对培养的细胞进行动态观察;并以免疫细胞化学技术对神经元进行染色。结果:在相差显微镜下可见加用N2添加剂和胎牛血清培养的神经元生长良好,胞体形态多样,突起数目多,长度长,与邻近神经元的胞体或突起形成接触,而胶质细胞则大量死亡,神经元纯化率达94%以上,未加用N2添加剂培养的神经元胶质细胞大量存在,神经元纯化率达50%左右。结论:N2添加剂和胎牛血清联合应用可使大鼠皮质神经元得到良好的生长和有效纯化。 相似文献
3.
目的观察含补阳还五汤的药物血清对体外培养大鼠脑皮层神经元生长的影响,为补阳还五汤治疗缺血性脑损伤提供实验依据。方法以体外培养新生SD乳鼠大脑皮层神经元为观察对象,采用四唑盐(MTT)比色法测定细胞生长情况。观察含药血清对常规培养及在缺氧状态下神经元生长的影响。结果补阳还五汤对常规培养及在缺氧条件下培养的神经元有显著促进生长作用,与空白血清组比较有显著差(P<0.05)。结论大鼠以补阳还五汤灌胃给药后,其血清中的药物成份有促进体外培养神经元细胞生长的作用。 相似文献
4.
卵巢未成熟畸胎瘤43例综合治疗疗效分析 总被引:3,自引:0,他引:3
为探讨不同手术方式及化疗方法对卵巢未成熟畸胎瘤的疗效及相关影响因素,对43例卵巢未成熟畸胎瘤患者进行回顾性分析。手术方式分为:保守性手术(患侧附件、大网膜切除术+盆、腹腔腹膜结节切除术+盆腔淋巴结清扫术)及根治性手术(全子宫、双附件及大网膜切除术_肿瘤细胞减灭术+盆腔淋巴结清扫术)。术后采用VAC(长春新碱,阿霉素,环磷酰胺)方案、BEP(博莱霉素或平阳霉素,足叶乙叉甙,顺铂或卡铂)方案或VBP(长春新碱,博莱霉素或平阳霉素,顺铂或卡铂)方案进行联合化疗。患者5年生存率根治性手术为44.4%,保守性手术为64.0%,差异无显著性。VAC方案与BEP(或VBP)方案的5年生存率分别为33.3%和77.3%(P<0.01)。疗程数≥4患者的5年生存率为73.9%,明显高于疗程数<4患者的38.9%(P<0.05)。认为对于有生育要求的卵巢未成熟畸胎瘤患者,可仅行保守性手术,术后辅以联合化疗。BEP(或VBP)方案对患者长期生存率的改善明显优于VAC方案。术后化疗的疗程数与疗效有关,对长期生存率的影响较明显。 相似文献
5.
P F Maness 《Journal of neuroscience research》1986,16(1):127-139
The advent of recombinant DNA technology has led to the identification in the DNA of normal animal cells of over 30 proto-oncogenes that are homologous to retroviral transforming genes. One of these encodes a protein kinase (pp60c-src) of unknown function, that is preferentially synthesized in brain and neural retina. Here the expression of pp60c-src in the peripheral nervous system was examined in sensory neurons from chick dorsal root ganglia with antisera raised against the transforming protein of Rous sarcoma virus (pp60v-src) expressed in Escherichia coli carrying the cloned v-src gene. This antiserum recognizes pp60c-src specifically in normal chicken cells. Western immunoblotting showed that dorsal root ganglia of stage 30 (day 6.5) chick embryos contained elevated levels of pp60c-src. Immunoperoxidase staining of neuron-enriched cultures prepared from chick dorsal root ganglia showed pp60c-src immunoreactivity in cells with neuronal morphology; flat, fibroblastic cells contained no detectable immunoreactivity. Indirect double immunofluorescence with pp60src antibodies and monoclonal antibodies against the 200-kD subunit of neurofilament protein confirmed that the cells expressing pp60c-src were neurons. Ninety-six percent of the neurofilament-positive cells were immunoreactive with pp60src antibodies, and conversely, all pp60c-src-positive cells were immunoreactive with neurofilament antibodies. pp60c-src immunofluorescence appeared to be distributed over the cell body, processes, and growth cones. These results clearly demonstrate that pp60c-src is a product of neurons and is expressed in sensory neurons in culture. 相似文献
6.
In vivo Intracellular Recording of Neurons in the Supraoptic Nucleus of the Rat Hypothalamus 总被引:3,自引:0,他引:3
R. E. J. Dyball J-G. Tasker J-P. Wuarin F. E. Dudek 《Journal of neuroendocrinology》1991,3(4):383-386
Intracellular recordings were made from cells in the hypothalamic supraoptic nucleus in the urethane-anaesthetized male rat using the ventral surgical approach. Impalements lasted from 5 min to 1 h and recorded cells had an input resistance of 55 to 170 megohms. Spikes of over 50 mV were recorded from 14 cells which could be antidromically activated by stimulation of the neural stalk. The spikes showed a hyperpolarizing afterpotential and the broadening characteristic of rapidly firing magnocellular neurons, which recovered rapidly (<200 ms). When depolarized, the cells showed evidence of a transient potassium current. Recurrent synaptic coupling between the recorded cell and adjacent cells would be expected to alter the hyperpolarizing afterpotential of an antidromic spike as compared with a spontaneous spike; no perceptible difference in the waveforms of the different types of spike could be detected in 11 spontaneously active cells. Application of just subthreshold stimuli to the neural stalk did not evoke depolarizing or hyperpolarizing potentials. Suprathreshold shocks to the neural stalk, when the antidromic spike was prevented by collision, also had no discernible effect on membrane potential. Thus intracellular recordings from magnocellular neurons in vivo revealed electrophysiological properties similar to those seen in vitro. No evidence for synaptic interconnection between magnocellular neurons was found in male rats. 相似文献
7.
幼兔缺血预处理在心肌保护第二窗期对未成熟心肌Bcl-2基因mRNA表达的影响 总被引:2,自引:1,他引:1
目的 观察幼兔心脏缺血预处理 2 4h对未成熟心肌缺血 /再灌注后Bcl -2mRNA表达的影响 ,探讨未成熟心肌保护第二窗形成的作用机理。方法 18只 3~ 4周龄幼兔 ,随机分为 3组 ,每组 6只 :Ⅰ组 ,正常对照组 ;Ⅱ组 ,缺血 /再灌注损伤组 ;Ⅲ组 ,缺血预处理组。建立活体心脏缺血 /再灌注损伤模型 ,Maclab/8s系统监测左心室发展压 (LVDP)、左室压上升及下降最大速率 (+dp/dtmax,-dp/dtmax)变化 ,原位免疫杂交 (ISH)方法检测心肌细胞内Bcl-2mRNA的表达。结果 缺血再灌注前 ,各组间LVDP、±dp/dtmax无显著性差异 (P >0 .0 5) ;再灌注 1h后 ,Ⅱ组的LVDP、+dp/dtmax和 -dp/dtmax恢复率分别为 (42 .81± 12 .56) %、(3 0 .67± 16.2 2 ) %和 (2 9.49± 10 .13 ) % ;Ⅲ组LVDP、+dp/dtmax及 -dp/dtmax恢复率分别为 (84.15± 13 .56) %、(69.49± 10 .3 6) %和 (58.3 7± 12 .45) % ,两组间有显著性差异 (P <0 .0 1)。Ⅱ组、Ⅲ组Bcl -2mRNA表达阳性细胞面积百分数分别为 (8.3± 2 .5) %、(76.3±13 .5) % ,两组间有显著性差异 (P <0 .0 1)。结论 缺血预处理可上调Bcl -2mRNA表达 ,参与未成熟心肌保护第二窗的形成 ,促进未成熟心肌缺血 /再灌注后心室功能恢复 相似文献
8.
Chemically distinct rat olivocochlear neurons. 总被引:6,自引:0,他引:6
We have produced a neurochemical map of the cell bodies of origin of the cochlear efferent terminals in rat by combining glutamic acid decarboxylase (GAD), choline acetyltransferase (ChAT), or calcitonin gene-related peptide (CGRP) immunocytochemistry with retrograde transport of horseradish peroxidase. The locations of cochlear efferent cell bodies are in general agreement with the medial and lateral systems described by White and Warr (J. Comp. Neurol. 219:203-214, 1983) with some minor modifications. The lateral system consists of at least two pools of chemically distinct neurons located within the lateral superior olive (LSO) ipsilateral to the injected cochlea. One pool immunostains with an antibody to GAD while the other immunostains with antibodies to ChAT and to CGRP. The medial efferent system consists of periolivary neurons that are almost exclusively large and ChAT-positive but CGRP-negative. They are located both ipsilateral and contralateral to the cochlea they project to. There are a few GAD-positive small neurons in the medioventral and rostral periolivary regions that project ipsilaterally, but these may prove tobe ectopic neurons. The ipsilateral lateroventral periolivary region (LVPO) contains some efferent neurons, all of which are ChAT-positive but CGRP-negative. Additional cochlear efferent neurons, some of which are ChAT-positive and others GAD-positive, are present within and immediately dorsal to the fiber capsule surrounding the medial limb, and to a lesser extent the lateral limb, of the ipsilateral LSO. Not all GAD-positive or ChAT-positive olivary cells project to the cochlea. We have complemented the results in the brainstem by demonstrating two immunocytochemically distinct populations of efferent terminals in the cochlea simultaneously, one CGRP-positive and the other GAD-positive. Approximately equal numbers of boutons immunoreactive for both markers are present beneath inner hair cells throughout the entire length of the cochlea. Surprisingly high numbers of GAD-positive and CGRP-positive boutons are also present on outer hair cells, with each class having its spatially and morphologically distinct features. The lack of CGRP-positive periolivary cells that are retrogradely labeled by cochlear injections of HRP suggests that the lateral olivocochlear system sends projections to outer hair cells. Our results raise questions about species differences in the organization of targets of the lateral and medial olivocochlear systems. 相似文献
9.
ZQ Yin MD SG Crewther PhD B Pirie BSc DP Crewther PhD 《Clinical & experimental ophthalmology》1997,25(4):107-109
Purpose: It was investigated whether alterations in neuronal structure and function occasioned by strabismic amblyopia also may be reflected in alterations in the expression on Y type neurons of a Cat-301 antibody sensitive antigen in the lateral geniculate nucleus (LGN) and cortex of our cat model of strabismic amblyopia. Methods/Results: The percentage of positively labelled cells was reduced in LGN laminae that received input from the deviated eye in strabismic amblyopic cats compared with normal cats. In the strabismic cortex, the density of immunopositive neurons was significantly reduced compared with normal, the effect being most pronounced in layer IV Conclusions: Despite previous physiological recordings indicating a decrease in X-cell associated acuity in strabismic amblyopia, the present findings imply that the changes in the early visual experience occasioned by strabismus also produce specific molecular changes in theY neuronal class. 相似文献
10.