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91.
Selective precipitation of ribonucleic acid from a mixture of total cellular nucleic acids extracted from cultured mammalian cells
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P. R. Harrison 《The Biochemical journal》1971,121(1):27-31
A simple and reproducible method is described for precipitating RNA selectively from total mammalian-cell nucleic acids extracted by the phenol-sodium dodecyl sulphate procedure at pH8.0. Under specified conditions bulk RNA is precipitated almost quantitatively whereas bulk DNA remains in solution. Minor components of RNA (detected by pulse-labelling and chromatography on methylated albumin-kieselguhr) and rapidly labelled components of DNA containing single-stranded regions are also precipitated. The usefulness of the method is discussed in the context of isolating separately both RNA and DNA from cultured cells that are difficult to obtain in quantity. 相似文献
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J. D. Harrison 《The Histochemical journal》1974,6(6):649-664
Synopsis The submandibular, sublingual and parotid glands of the cat have been studied. Mucosubstance histochemistry demonstrated acidic mucosubstances with varying properties in the acini. Thiamine pyrophosphatase and nucleoside diphosphatase reaction products were seen with a Golgi-like appearance in acinar cells. Granules of acid phosphatase, -glucuronidase and E600-resistant esterase reaction products, presumably representing lysosomal enzyme activities, were seen in acinar and ductal cells. Diffuse acid phosphatase and -glucuronidase reaction products were seen in central cells of the submandibular acini, and diffuse non-specific esterase reaction product was seen in acinar and ductal cells. Arylamidase reaction product was associated with some acinar cells. Reaction product from a peroxidase technique was seen in demilunar cells of the submandibular acini, in parts of the sublingual acini, in parotid acini, and in ductal cells. Cytochrome oxidase and succinate dehydrogenase reaction products were seen most strongly in striated ducts, whereas NADH- and NADPH-diaphorase reaction products were seen at a high level throughout the ducts. 相似文献
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S-(+)-3,4-Dihydroxybutylphosphonic acid, an isosteric analogue of sn-glycerol 3-phosphate, was synthesized stereospecifically and shown to be an effective substrate for rabbit muscle glycerol 3-phosphate dehydrogenase (sn-glycerol 3-phosphate-NAD(+) oxidoreductase, EC 1.1.1.8). Non-isosteric phosphonate analogues of sn-glycerol 3-phosphate showed neither substrate nor inhibitory activity with the enzyme. 相似文献
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Microbiological evaluation of Pacific shrimp (Pandalus jordani) processing was made from samples obtained at five key processing points. The microbial count of raw shrimp ranged from 1.3 x 10(6) to 3.0 x 10(6). The initial microbial flora, in order of predominance, was Acinetobacter-Moraxella, Flavobacterium, Pseudomonas, gram-positive cocci, and Bacillus species. No yeasts were isolated. Differences in processing practices influenced both microbial count and the shrimp flora. The microbial load, however, always increased after peeling and sorting operations and decreased after cooking, washing, and brining steps. Significantly, the gram-positive cocci were recovered with increasing frequency after each processing step, reaching 76% of the total load in a final product. Most of them, however, were coagulase-negative. 相似文献
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Summary Salivary myoepithelial cells were demonstrated by alkaline-phosphatase techniques in cat, but not in man or dog, and by an adenosine-triphosphatase technique in man, but not in cat or dog.Electron-microscopical cytochemistry showed that the reaction product from the respective techniques in cat and man was associated with the myoepithelial plasma membrane and that it was most constant and usually strongest at the plasma membrane adjacent to the acinar cells.In the dog, the reaction product from the adenosine-triphosphatase technique was found lining the canaliculi and lumina of the acini of the parotid gland, and of the non-mucous acini of the submandibular and sublingual glands. Alkaline-phosphatase reaction product was found lining the canaliculi and lumina in the sublingual gland.These remarkable species differences indicate that neither technique can be regarded as a universal marker of salivary myoepithelial cells. Inconsistencies in activity were found within the myoepithelium of individual glands and suggest that even the appropriate technique may not be relied upon to demonstrate all the myoepithelial cells present in a tissue section.
This work has been supported by a Medical Research Council Grant. 相似文献
Zusammenfassung Myoepithelzellen der Speicheldrüsen lassen sich bei der Katze — nicht jedoch bei Mensch und Hund — mittels der alkalischen Phosphatase-Reaktion darstellen. Der Nachweis für Adenosintriphosphatase fällt in diesen Zellen beim Menschen, nicht bei Katze und Hund, positiv aus.Elektronenmikroskopisch-cytochemisch zeigt sich, daß das Reaktionsprodukt der jeweiligen Methode bei der Katze und beim Menschen an den Plasmamembranen der Myoepithelzellen auftritt und zwar am regelmäßigsten und meistens am stärksten in der Nachbarschaft der Azinuszellen.Beim Hund fällt in der Ohrspeicheldrüse sowie den nicht-mukösen Acini der Glandulae submandibularis und sublingualis das Reaktionsprodukt der Adenosintriphosphatase-Reaktion an der Begrenzung der Azinuskanälchen und der Lumenoberfläche aus. Alkalische Phosphatase ist in der Glandula sublingualis in den Wänden der Kanälchen und an den Lumina lokalisiert.Diese bemerkenswerten Unterschiede zwischen den verschiedenen Tierarten zeigen, daß keine der verwendeten Methoden zur Universalmarkierung von Myoepithelzellen der Speicheldrüsen geeignet ist. Außerdem ist die Aktivität des Myoepithels bei den einzelnen Drüsen uneinheitlich. Dies legt die Vermutung nahe, daß man sich selbst bei Anwendung der richtigen Methode nicht darauf verlassen kann, alle in einem Gewebsschnitt vorhandenen Myoepithelzellen zu erfassen.
This work has been supported by a Medical Research Council Grant. 相似文献
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