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111.
目的 探讨获取母血中纯化胎儿细胞的有效方法。方法 对 6 0名孕龄 7~ 40周、年龄 2 1~ 30岁的孕妇外周血进行不连续密度梯度离心 ,将分离后的细胞进行制片 ,显微镜下行显微操作分离胎儿有核红细胞 ,进行 Y染色体特异性DYZ1基因的聚合酶链反应 (PCR)。结果 (1)早孕期母血中胎儿 NRBC检出率为 2 0 % ,中孕期母血中胎儿 NRBC检出率为 71.4% ,晚孕期母血中胎儿 NRBC检出率为 6 .6 5 % ,三者之间有显著性差异 (P <0 .0 5 ) ;(2 )用显微操作技术获得的胎儿细胞数量已满足 PCR扩增所需的模板量 ,对胎儿性别进行预测 ,男胎符合率为 11/ 13(84.6 % ) ,性别符合率为 2 6 / 2 8(92 .9% )。结论 不连续密度梯度离心合并显微操作技术对于无创性早期产前基因诊断意义重大。 相似文献
112.
A. Prelle G. Fagiolari N. Checcarelli M. Moggio A. Battistel G.P. Comi P. Bazzi A. Bordoni M. Zeviani G. Scarlato 《Acta neuropathologica》1994,87(4):371-376
In situ hybridization combined with immunohistochemical techniques has been applied to study patients affected by mitochondrial myopathies with large mitochondrial (mt)DNA deletions. All patients' muscle biopsies showed ragged red fibers (RRFs) and cytochrome oxidase (COX) deficiency. Two digoxygenin-labeled, polymerase chain reaction (PCR)-amplifed DNAs were used as probes. One probe was designed to hybridize only with wild-type mtDNAs, while the other recognized both wild-type and deleted mtDNAs. Concomitant immunocytochemical analysis using antibodies against subunits II, III, (encoded by mtDNA) and IV (encoded by nuclear DNA) of COX was carried out. In our patients deleted mtDNAs are overexpressed in COX-negative RRFs, while wild-type mtDNAs are decreased in the same fibers. Immunohistochemistry studies show that COX IV is overexpressed in RRFs and that COX II and COX III subunits are still present. Deleted mtDNAs are spatially segregated in muscle fibers, where they interfere with the local population of normal mitochondrial genomes, causing a regional deficiency of the mitochondrial respiratory activity.This work was supported by the Associazione Amici del Centro Dino Ferrari 相似文献
113.
Imamura Hitoshi Maruyama Toru Okabe Hiroaki Shimada Hidekata Otagiri Masaki 《Pharmaceutical research》1994,11(4):566-570
We examined different fluorescent probes suitable for fluorometric determination of 1-acid glycoprotein (AGP) in serum. Quinaldine red (QR) was shown to bind strongly and selectively to AGP. Taking advantage of the enhanced fluorescence of QR in the presence of AGP, we developed a direct method for the determination of serum AGP without removal of other serum proteins such as albumin. AGP concentrations in serum of healthy volunteers and patients correlated well with results from the conventional single radial immunodiffusion (SRID) method (r = 0.93, slope = 1). The newly developed method is faster and has a larger analytical concentration range than the SRID method. This method can also be used to determine AGP in serum of experimental animals, and it can serve to monitor AGP serum concentrations for pharmacokinetic evaluation of basic drugs. 相似文献
114.
The direct-current electroretinogram of seven pigmented and seven albino rabbits was recorded from both eyes for almost 4 h in response to repeated identical light stimuli. Stimulus duration was 10 s, light intensity was 6.8 × 102 lux, and the interval between the beginning of succeeding light stimuli was 3 min. The dark-adaptation period preceding light stimulation was 30 min for one of the eyes (unoccluded eye) and 150 min for the contralateral eye (occluded eye), which was patched during the first part (117 min) of the experiment. In pigmented animals, the b- and c-wave amplitudes of the unoccluded eye slowly increased during the first part of the experiment but not significantly during the second. The a-wave amplitude was not significantly changed. After removal of the cover, the a- and b-wave amplitudes of the occluded eye immediately attained but did not exceed the level of those in the unoccluded eye, irrespective of the light adaptation induced by the stimulus flashes previously presented to the unoccluded eye. (Control experiments on six pigmented rabbits confirmed that stimuli identical to those used in the main part of the study caused a light adaptation, since a decrease in a- and b-wave amplitudes occurred after the first light stimulus following an initial dark-adaptation period of 2 h for both eyes.) In albino rabbits, electroretinogram responses were clearly discernible in the occluded eye also during the first part of the experiment, probably because of transillumination of the head. In other respects, the results were essentially similar to those of pigmented animals. The observation that occluded eyes did not dark adapt better, as judged by the electroretinogram responses, than contralateral eyes given repeated light adaptive stimuli may indicate the presence of a mechanism for transfer of adaptation information between the eyes. 相似文献
115.
Ragnheiđur Bragadóttir Sven Jarkman 《Documenta ophthalmologica. Advances in ophthalmology》1995,90(3):291-303
The effects of the stable cyclic adenosine monophosphate analogue adenosine 3, 5-cyclic monophosphorothioate Sp-isomer (Sp-cAMPS) on the direct-current electroretinogram and the standing potential of the eye were studied. Corneal recordings were obtained from unilaterally vitrectomized albino rabbit eyes during alternating intravitreal perfusions with Sp-cAMPS and a control solution (Pharmacia eye irrigating solution). The contralateral eye was used as a control. To evaluate further the effects on the c-wave,in vivo intraretinal microelectrode measurements were made during simultaneous intravitreal perfusion of Sp-cAMPS and irrigating solution, respectively. Sp-cAMPS in concentrations of 1, 10 and 100µM was tested by corneal direct-current electroretinography. There was no significant effect on the a-wave amplitude. The b-wave amplitude was reversibly elevated at an Sp-cAMPS concentration of 100µM (p<0.01, n=7). The c-wave amplitude was reversibly elevated at a concentration of 10µM (p<0.001, n=8), and this effect was more pronounced at 100µM (p<0.001, n=7). The SP increased reversibly at a concentration of 100µM (p<0.001, n=7). Microelectrode recordings were performed with Sp-cAMPS at a concentration of 100µM. The recordings showed significant increases in both the transepithelial potential (p<0.01, n=3) and the slow PIII (p<0.01, n=3). The effects of Sp-cAMPS on the b-wave as well as on the two components of the c-wave suggest influences on both the inner retina and the retinal pigment epithelium of the rabbit eye.Abbreviations PHS
Pharmacia eye irrigating solution
- AMP
adenosine monophosphate
- Sp-cAMPS
adenosine 3, 5 - cyclic monophosphorothioate Sp-isomer 相似文献
116.
117.
Immunocytochemical Study of Cells in the Vitreous of Proliferative Virteoretretinopathy 总被引:1,自引:0,他引:1
Purpose: To identify the cellular components of vitreous samples obtained during vit-rectomy for proliferative vitreoretinopathy(PVR).Methods: With the use of three intermediate filament (IF) proteins, vimentin, glialfibrillary acidic protein (GFAP), and cytokeratin (CK), cytocentrifuge slides of 14fresh vitreous aspirates were detected with immunohistochemical technique.Results: All the specimens contained epithelial-like proliferative cells with or withoutpigment and some membrane-like pieces. Immunocytochemical staining showed that76.0-90.0% cells stained for CK, 17.4-29.6% cells expressed GFAP, and 80.1-91.0% cells were positive for vimentin.Conclusions : Majority of cells in the vitreous samples originated from retinal pigmentepithelial cells (RPE) and glial cells in PVR. Expression of IF proteins may be determinedby tissue of origin and local microenvironment. Eye Science 1999 ; 15; 13 - 16. 相似文献
118.
R. Bültmann Marie Trendelenburg Florin Tuluc Henning Wittenburg Klaus Starke 《Naunyn-Schmiedeberg's archives of pharmacology》1999,359(4):339-344
In order to assess the consequences of a concomitant blockade of P2X-receptors and ecto-nucleotidases, effects of 13 P2-receptor
antagonists were investigated on contractions of the rat vas deferens elicited by α,β-methylene ATP (α,β-MeATP) and ATP and
on the removal of ATP from the incubation medium by vas deferens tissue.
Increasing concentrations of all antagonists reduced and finally abolished contractions elicited by α,β-MeATP (3 μM), with
IC50-values ranging from 1.1 to 100 μM. Pyridoxalphosphate-6-azophenyl-2’,4’-disulphonate (PPADS), 6-azophenyl-4-amino-5-hydroxy-naphthalene-1,3-disulphonate
(NH02), 4,4’-diisothiocyanatostilbene-2,2’-disulphonate (DIDS) and uniblue A also progressively reduced and finally abolished
contractions elicited by ATP (1 mM). 8,8’-[Carbonylbis(imino-3,1-phenylenecarbonyl-imino)]-bis-(1,3,5-naphthalenetrisulphonate)
(NF023), sura- min, pyridoxalphosphate-6-azophenyl-2’,5’-disulphonate (iso-PPADS), trypan blue and reactive blue 19, in contrast,
caused only partial blockade, by 34–43% maximally; reactive blue 2 and reactive red 2 had no effect; and 6,6’-(1,1’-biphenyl-4,4’-diylbisazo)-bis-4-amino-5-hydroxy-naphtha-lene-1,3-disulphonate
(NH01) and Evans blue even enhan- ced the response to ATP. For antagonists causing full or partial inhibition, the IC50-values against ATP were close to those against α,β-MeATP. All antagonists attenuated the removal of ATP, with IC25%-values ranging from 0.8 μM to >320 μM.
The results confirm the frequent combination, in one antagonist molecule, of P2-receptor blockade and blockade of ecto-nucleotidases.
This dual action underlies the effect of such compounds on contractions of the vas deferens elicited by ATP which, for certain
substances (e.g., suramin, reactive blue 2), can be explained by a simple model in which the antagonist simultaneously blocks
the degradation of ATP and a single contraction-mediating receptor (P2X1). Several observations, however, do not conform with this model, and the existence of multiple contraction-mediating receptors
for ATP or multiple, pharmacologically distinct ecto-nucleotidases has to be considered.
Received: 23 October 1998 / Accepted: 11 January 1999 相似文献
119.
120.
Pentose shunt activity in developing chick retina and pigment epithelium was studied by measuring the rate of 14CO2 evolution from glucose selectively labelled in the C-1 and C-6 positions. In the retina, shunt activity declines from appreciable levels at stages 29–31 to minimal activity in the 2-week-old hatched chick. Overall retinal metabolism also declines up to stage 45, but dramatically increases again after hatching. Developing chick pigment epithelium has minimal shunt activity at all stages studied. In contrast, cultured chick pigment epithelium has appreciable shunt activity which is constant over a period of several weeks in culture. This appears to be a switch in biochemical differentiation which could form the basis at least in part for subsequent changes in cell types observed in cultured pigment epithelial cells byEguchi and Okada (1973). 相似文献