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141.
Neural crest cells are motile and mitotic, whereas their neuronal derivatives are terminally post-mitotic and consist of stationary cell body from which processes grow. The present study documents changes in the cytoskeleton that occur during neurogenesis in cultures of avain neural crest cells. The undifferentiated neural crest cells contain dense bundles of actin filaments throughout their cytoplasm, and a splayed array of microtubules attached to the centrosome. In newly differentiating neurons, the actin bundles are disrupted and most of the remaining actin filaments are reorganized into a cortical layer underlying the plasma membrane of the cell body and processes. Microtubules are more abundant in newly-differentiating neurons than in the undifferentiated cells, and individual microtubules can be seen dissociated from the centrosome. Neuron-specific beta-III tubulin appears in some crest cells prior to cessation of motility and cell division, and expression increases with total microtubule levels during neurogenesis. To investigate how these early cytoskeletal changes might contribute to alterations in morphology during neurogenesis, we have disrupted the cytoskeleton with pharmacologic agents. Microfilament disruption by cytochalasin immediately arrests the movement of neural crest cells and causes them to round-up, but does not significantly change the morphology of the immature neurons. Microtubule depolymerization by nocodazole slows the movement of undifferentiated cells and causes retraction of processes extended by the immature neurons. These results suggest that changes in the actin and microtubule arrays within neural crest cells govern distinct aspects of their morphogenesis into neurons. 相似文献
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MA Reina Perticone A López Garciá ME Gorra J Daneri 《Canadian Metallurgical Quarterly》1998,64(11):489-497
BACKGROUND AND AIM: Several authors have focused on a causal link between the onset of neurological complications after lumbar injections and the fact that epithelial cells may be drawn into the vertebral canal during these procedures. Complications may arise both early (cephalea, septic and aseptic meningitis) and late (epidermoid tumours). The authors aimed to evaluate whether skin fragments which are carried down by the needle during subarachnoid anesthesia may even be present in the epidural or subarachnoid space three days later and may therefore justify the onset of the above neurological syndromes. METHODS: Five adult cats under narcosis underwent subarachnoid anesthesia using disposable 22G Quincke type needles. Between 0.7 and 1 ml isobaric bupivacaine at 0.50% was injected. The presence of the motor block of the lower limbs was ascertained once the effects of general anesthesia wore off. On the third day, again under general anesthesia, cardio-respiratory arrest was provoked by intravenous injection. Samples of meninges were collected in the injection area. After fixation in a phosphate glutaraldehyde buffer, dehydration in acetone, dehydration by critical point and gold metalisation, the samples were examined using SEM. RESULTS: No epidermal cells were found on the surface of the meninges. On the other hand, a squamous epithelial cell was observed which drained inside a sectioned epidural vessel towards the systemic circulation. CONCLUSIONS: This study confirms the possibility that, after subarachnoid anesthesia using 22G Quincke needles, skin fragments may enter the spinal canal. The permanence or otherwise of the epithelial fragments on the third day depends on the size of the fragment drawn down and the efficacy of the drainage system which removes isolated epithelial cells. This phenomenon may justify the self-limiting character of cephalea and meningisms which, even if not treated, regress in a few days, as well as the scarce development of epidermoid tumours. 相似文献
147.
JS Plummer KA Berryman C Cai WL Cody J DiMaio AM Doherty JJ Edmunds JX He DR Holland S Levesque DR Kent LS Narasimhan JR Rubin ST Rapundalo MA Siddiqui AJ Susser Y St-Denis PD Winocour 《Canadian Metallurgical Quarterly》1998,8(23):3409-3414
The synthesis and antithrombotic activity of a series of nonpeptide bicyclic thrombin inhibitors is described. We have explored the SAR with modifications to the P1 site. The introduction of arginine mimetics at the P1 site led to potent and selective thrombin inhibitors. 相似文献
148.
GL Alonso DA González D Takara MA Ostuni GA Sánchez 《Canadian Metallurgical Quarterly》1998,1405(1-3):47-54
The sarcoplasmic reticulum Ca-ATPase is fully activated when approximately 1 microM [Ca2+] saturates the two transport sites; higher [Ca] inhibits the ATPase by competition of Ca-ATP with Mg-ATP as substrates. Here we describe a novel effect of EGTA and other chelators, raising the possibility of an additional activating effect of Ca in the sub- or low microM range. Sarcoplasmic reticulum membranes were isolated from rabbit skeletal muscles. The ATPase activity was measured after incubation at 37 degreesC in 3 mM ATP, 3 mM MgCl2, 50 mM MOPS-Tris (pH 7.2), 100 mM KCl, and variable CaCl2, EGTA and calcimycin. In the absence of added EGTA and Ca the ATPase activity is high due to contaminant Ca. The determination of the ATPase activity in the presence of increasing amounts of EGTA, without added Ca, yields a decreasing sigmoidal function. Ki ranged between 20 and 100 microM, depending on the enzyme concentration. Pi production is linear with time for several [EGTA] yielding suboptimal ATPase activities, which are inhibited by thapsigargin. These suboptimal Ca-ATPase activities are inhibited by preincubation of the enzyme in EGTA, at pH 7.2. This effect increases upon increasing EGTA concentration and preincubation time. The inhibitory effect of the previous exposure of the enzyme to EGTA is partially but significantly reverted by increasing [Ca2+] during incubations. Calcimycin and EDTA have similar effects as EGTA when added in preincubations. The effect of calcimycin is fully reverted by optimal [Ca2+] in incubations. The effects of EGTA, EDTA and calcimycin in preincubation are not additive. The results suggest that an additional calcium, lost during preincubations from a site with affinity near 1 microM, is necessary for full activation of the ATPase. 相似文献
149.
E Cuende V Almeida J Portu M Aldamiz MA Erdozain JC Vesga N Saracibar 《Canadian Metallurgical Quarterly》1998,41(10):1884-1888
We report the case of a 27-year-old patient with the human immunodeficiency virus (HIV) infection who presented with a 2-week history of crops of painful, red papules over the trunk and extremities, together with a sterile, symmetric polyarthritis involving the small and large joints. Histologic study of a skin biopsy specimen demonstrated features of papulonecrotic tuberculid. Analytical and microbiologic studies ruled out tuberculous infection. Both the synovial and the skin processes were considered to be an immune response secondary to Mycobacterium tuberculosis infection. Specific treatment was established, and there was marked improvement in both the skin and joint symptoms. This case illustrates the complex relationship between the host and the HIV, suggesting an immune dysregulation cause for both the synovial and the skin lesions. 相似文献
150.
BA Posner MB Mixon MA Wall SR Sprang AG Gilman 《Canadian Metallurgical Quarterly》1998,273(34):21752-21758
Agonist-bound heptahelical receptors activate heterotrimeric G proteins by catalyzing exchange of GDP for GTP on their alpha subunits. In search of an approximation of the receptor-alpha subunit complex, we have considered the properties of A326S Gialpha1, a mutation discovered originally in Gsalpha (Iiri, T., Herzmark, P., Nakamoto, J. M., Van Dop, C., and Bourne, H. R. (1994) Nature 371, 164-168) that mimics the effect of receptor on nucleotide exchange. The mutation accelerates dissociation of GDP from the alphai1beta1gamma2 heterotrimer by 250-fold. Nevertheless, affinity of mutant Gialpha1 for GTPgammaS is high in the presence of Mg2+, and the mutation has no effect on the intrinsic GTPase activity of the alpha subunit. The mutation also uncouples two activities of betagamma: stabilization of the GDP-bound alpha subunit (which is retained) and retardation of GDP dissociation from the heterotrimer (which is lost). For wild-type and mutant Gialpha1, beta gamma prevents irreversible inactivation of the alpha subunit at 30 degreesC. However, the mutation accelerates irreversible inactivation of alpha at 37 degreesC despite the presence of beta gamma. Structurally, the mutation weakens affinity for GTPgammaS by steric crowding: a 2-fold increase in the number of close contacts between the protein and the purine ring of the nucleotide. By contrast, we observe no differences in structure at the GDP binding site between wild-type heterotrimers and those containing A326S Gialpha1. However, the GDP binding site is only partially occupied in crystals of G protein heterotrimers containing A326S Gialpha1. In contrast to original speculations about the structural correlates of receptor-catalyzed nucleotide exchange, rapid dissociation of GDP can be observed in the absence of substantial structural alteration of a Galpha subunit in the GDP-bound state. 相似文献