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121.
122.
In vitro growth and differentiation of human kidney tubular cells on a basement membrane substrate 总被引:4,自引:0,他引:4
An Hang Yang Jeanine Gould-Kostka Terry D. Oberley 《In vitro cellular & developmental biology. Plant》1987,23(1):34-46
Summary Kidney cortical tubular cells, mainly proximal tubular cells, isolated from human kidney and grown either on a basement membrane
substrate in chemically defined medium or on plastic in serum-supplemented medium, had substantial proliferative potential
and could be propagated for more than 10 generations or 8 passages before senescence. Basement membrane produced on a plastic
substrate by the HR-9 endodermal cell line could replace serum supplementation in promoting tubular cell growth. Tubular cells
grown on an HR-9 basement membrane substrate exhibited stable epithelial morphology over an extended period of time; in the
presence of 5% serum they differentiated into organized structures such as hemicysts and cell cords. Cells grown on plastic
failed to differentiate and gradually degenerated. Tubular cells on HR-9 basement membrane were characterized by densely packed
microvilli, abundant rough endoplasmic reticulum and free polysomes, basal cell membrane interdigitations, a well-developed
endocytotic apparatus, and conspicuous junctional complexes—all features of the proximal tubular cell. Compared with cells
on plastic substrate, there were higher levels of the brush border enzymes γ-glutamyl transpeptidase,l-leucine aminopeptidase, and alkaline phosphatase in cells maintained on an HR-9 basement membrane substrate, further supporting
the conclusion that a basement membrane substrate promoted differentiation of tubular cells. These data and morphological
observations indicate that a basement membrane substrate can promote growth and both functional and morphologic differentiation
of human kidney tubular cells.
This work was supported by the Veterans Administration. 相似文献
123.
Kidney glomerular explants in serum-free media. Sequential morphologic and quantitative analysis of cell outgrowths 总被引:3,自引:0,他引:3
T D Oberley B W Steinert A H Yang P J Anderson 《Virchows Archiv. B, Cell pathology including molecular pathology》1986,50(3):209-235
Guinea pig glomeruli were grown in vitro for 22 days in a serum-free medium composed of Waymouth's MB 752/1 supplemented with sodium pyruvate, nonessential amino acids, antibiotics, insulin, transferrin, selenium, triiodothyronine, and fibronectin (FN), and sequential morphologic and quantitative studies of cell outgrowth were performed. Glomeruli grown in serum-free medium showed preservation of glomerular visceral epithelial cells but extensive necrosis of endocapillary cells (endothelial and mesangial cells). Morphologic analysis demonstrated progressive morphologic changes in cultured glomerular cells; however, most cell types observed in culture appeared to grow from the epithelial side of the glomerular basement membrane. Mitosis was a prominent component of glomerular cell outgrowth in vitro, and total DNA increased slightly during glomerular culture. FN was required for glomerular cell outgrowth, and studies using FN fragments demonstrated that the carboxy-terminal portion of FN was required for whole glomerular attachment. These results are used to develop a model for glomerular cell outgrowth in vitro. 相似文献
124.
Kaiyu Yang 《World journal of microbiology & biotechnology》1986,2(4):421-426
Summary Studies on immobilized enzymes and whole cells have been made since 1970 in China. A new method for immobilization was developed by using the bifunctional reagent, -sulphatoethyl-sulphonylaniline to activate polysaccharides. More than ten immobilized biocatalysts such as polynucleotide phosphorylase and penicillin acylase, have been used in the pharmaceutical industry, the production of high value chemicals, and in pollutant treatment. The prospects and scientific constraints in this field are discussed in this paper.
Paper presented at the VII International Conference on the Global Impacts of Applied Microbiology, Helsinki, 12–16 August 1985. 相似文献
Resumen Desde 1970 se estan llevando a cabo en China estudios sobre enzimas inmovilizados y células enteras. Un nuevo método para la inmovilización fue diseñado utilizando el reactivo bifuncional -sulfatoetilsulfonilanilina para activar polisacaridos. Màs de 10 biocatalizadores inmovilizados, tales como polinucleotidofosforilasa y penicillin-acilasa, se han utilizado en la industria farmaceútica, en la producción de compuetos químicos de alto coste y en el tratamiento de polucionantes. En este trabajo se discuten las perspectivas y las dificultades científicas en este campo.
Résumé Des études sur les enzymes fixés et les cellules immobilisées ont été entreprises en Chine à partir de 1970. II a été mis au point une nouvelle méthode d'immobilisation, utilisant pour l'activation des polysaccharides un réactif bifonctionnel, la -sulphato-éthyl-sulphonyl-aniline. Plus de 10 biocatalyseurs immobilisés, dont la polynucléotide-phosphorylase et la pénicilline-acylase, ont été employés dans l'industrie pharmaceutique, pour la production de composés chimiques à haute valeur commerciale et pour la dépollution. Les perspectives et les contraintes scientifiques dans le domaine considéré sont discutées.
Paper presented at the VII International Conference on the Global Impacts of Applied Microbiology, Helsinki, 12–16 August 1985. 相似文献
125.
A series of sequential polypeptides (LysiRj)n (R is Leu, Ser, or Gly) and random copolypeptides, (Lysx, Leuy)n, were synthesized. Their conformation in NaDodSO4 solution was determined by CD. Only (Lys-Leu)n, (Lys-Ser)n, and (Lys3-Ser)n adopt a stable β-form in the surfactant solution; (Lys-Ser2)n, (Lys-Ser3)n, (Lys2-Ser2)n, and (Lys2-Ser)n have an unstable β-form, which reverts to an unordered form in high NaDodSO4 concentrations, even though both Ser and DodSO-bound Lys+ are β-formers. In contrast, (Lys-Gly)n remains unordered in NaDodSO4 solution. On the other hand, Lys-rich (Lys2-Leu)n forms an unstable helix and (Lys2-Leu2)n a stable helix in NaDodSO4 solution. In 25 mM NaDodSO4 (Lysx, Leuy)n also forms a helix up to x = 75 and reverts to the β-form at x = 90. This compares with the helical conformation of (Lysx, Alay)n up to x = 65 and its β-form at x = 90, suggesting that Leu is an even stronger helix-former than Ala. Our results may provide a plausible explanation for the increase in helicity and disruption of the β-form for many proteins in NaDodSO4 solution, that is, the polypeptide chain of a protein usually favors a helical conformation over a β-form in the presence of excess surfactant. 相似文献
126.
Preparation and performance of immobilized yeast cells in columns containing no inert carrier 总被引:1,自引:0,他引:1
Schizosaccharomyces pombe was cultivated in a medium of glucose (10 g/L) malt extract (3 g/L), yeast extract (3 g/L), and bactopeptone (5 g/L) to form flocs. More than 95% of the cell population were flocculated. Variation in glucose concentration (from 10 to 100 g/L) did not affect flocculation. Yeast extract helped induce flocculation. Application of the immobilized yeast for the continuous production of ethanol was tested in a column reactor. Soft yeast flocs (50-200 mesh) underwent morphological changes to heavy particles (0.1-0.3 cm diameter) after continuously being fed with fresh substrates in the column. Productivity as high as 87 g EtOH L(-1) h(-1) was obtained when a 150 g/L glucose medium was fed. The performance of this yeast reactor was stable over a two-month period. The ethanol yield was 97% of the theoretical maximum based upon glucose consumed. 相似文献
127.
Reversal of Fv-1 host range by in vitro restriction endonuclease fragment exchange between molecular clones of N-tropic and B-tropic murine leukemia virus genomes. 总被引:10,自引:10,他引:0 下载免费PDF全文
L R Boone F E Myer D M Yang C Y Ou C K Koh L E Roberson R W Tennant W K Yang 《Journal of virology》1983,48(1):110-119
We molecularly cloned unintegrated viral DNA of the BALB/c endogenous N-tropic and B-tropic murine leukemia retroviruses and in vitro passaged N-tropic Gross (passage A) murine leukemia retroviruses. Recombinant genomes were constructed in vitro by exchanging homologous restriction enzyme fragments from N- or B-tropic parents and subsequent recloning. Infectious virus was recovered after transfection of these recombinant genomes into NIH-3T3 cells and cocultivation with the Fv-1 nonrestrictive SC-1 cells. XC plaque assays of recombinant virus progeny on Fv-ln and Fv-lb cells indicated that the Fv-l host range was determined by sequences located between the BamHI site in the p30 region of the gag gene (1.6 kilobase pairs from the left end of the map) and the HindIII site located in the pol gene (2.9 kilobase pairs from the left end of the map). 相似文献
128.
The effect of X rays on the reproductive death of cultured normal human mammary epithelial cells was examined. Techniques were developed for isolating and culturing normal human mammary epithelial cells which provide sufficient cells at second passage for radiation studies, and an efficient clonogenic assay suitable for measuring radiation survival curves. It was found that the survival curves for epithelial cells from normal breast tissue were exponential and had D0 values of about 109-148 rad for 225 kVp X rays. No consistent change in cell radiosensitivity with the age of donor was observed, and no sublethal damage repair in these cells could be detected with the split-dose technique. 相似文献
129.
R S Slesinski P J Guzzie W C Hengler P G Watanabe M D Woodside R S Yang 《Mutation research》1983,124(3-4):299-314
Ethylenediamine (EDA) was evaluated for potential genotoxic activity using a battery in vitro and in vivo mammalian tests. The tests employed were the Chinese hamster ovary (CHO) gene mutation assay, the sister-chromatid exchange (SCE) test with CHO cells, unscheduled DNA synthesis (UDS) assays with primary rat hepatocytes and a dominant lethal study with Fischer 344 rats. EDA did not produce a positive, dose-related, mutagenic effect in either the CHO mutation assay or in the SCE test when evaluated both with and without the addition of a rat-liver S9 activation system. With hepatocytes, no positive effects of EDA upon UDS values were noted in 2 separate studies using either a scintillation counting procedure or an autoradiographic method to determine UDS activity. In a dominant lethal study, male rats fed for 23 weeks with dietary levels of EDA X 2HCl of 0, 0.05, 0.15 or 0.50 g/kg/day, and mated with 1 virgin female/week for 3 consecutive weeks, showed no dose-related or statistically significant effects upon fertility, total number of implantations/female, or the number of living and dead implants per female; marked effects upon the incidence of dominant lethal mutations were noted in the positive control group injected intraperitoneally with one dose of 0.25 mg/kg triethylenemelamine. We conclude that EDA was not genotoxic in the in vitro and in vivo mammalian test systems employed. 相似文献
130.
Meloidogyne enterolobii n. sp. is described and illustrated from roots of pacara earpod tree, Enterolobium contortisiliquum (Vell.) Morong, on Hainan Island in China. The perineal pattern of the female is usually oval shaped, the striae are fine to coarse, the dorsal arch is moderately high to high and usually rounded, and the phasmids are large. The stylet knobs in females are divided longitudinally by a groove so that each knob appears as two. The mean distance of the excretory pore to the anterior end in the female is 62.9 μm. Males have a large, rounded labial disc that fuses with the medial lips to form a dorso-ventrally elongate head cap. The labial disc is slightly elevated, and the medial lips are crescent shaped. The second-stage juvenile mean body length is 436.6 μm. The lateral lips are large and triangular in face view. The tail is 56.4 μm long and narrow with a broad, bluntly rounded tip. M. enterolobii n. sp reproduces well on E. contortisiliquum and causes severe damage. Other good hosts include cotton, resistant tobacco ''NC 95,'' pepper, watermelon, and tomato. 相似文献