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71.
72.
alpha-Endorphin and gamma-endorphin, two closely related peptides of the pro-opiomelanocortin family with characteristic biological activities, were purified to homogeneity from single human pituitary glands and chemically identified. Isolation of the peptides was based on size fractionation by Sephadex G-75 chromatography followed by two HPLC steps using reverse-phase and paired-ion reverse-phase systems and was monitored by radioimmunoassay. During the isolation procedure alpha- and gamma-endorphin-sized material behaved chromatographically and immunologically indistinguishably from synthetic alpha- and gamma-endorphin. The amino acid composition and NH2-terminus of isolated peptides demonstrated their identity as authentic alpha-endorphin and gamma-endorphin. Acetylated forms were absent. In addition, evidence is provided that large forms with alpha- and gamma-endorphin immunoreactivity detected during gel filtration are human lipotropin-(1-74) and -(1-75), respectively. The data substantiate that alpha-endorphin and gamma-endorphin exist as endogenous peptides in the human pituitary gland. 相似文献
73.
Nervous system tumors are one of the leading causes of cancer related death. Specific mechanisms facilitating the invasive
behavior of gliomas remain obscure. Advanced simulation models of the in vivo response to therapy conditions should potentially
improve malignant glioma treatment. Expressional profiling of vimentin––one of reliable pro-invasive tumor makers––in those
simulation models was the goal of this study, in order to estimate a pro-invasive response of surviving malignant glioma cells
under clinically relevant therapeutic conditions. Human U87-MG malignant glioma cells were used. These cells are characterized
by the wild p53-phenotype, which is relevant for the majority of primary malignant glioblastomas. Experimental design foresaw the cells to
undergo either irradiation or chemo-treatment with temozolomide alone, or combined treatment. Expression profiling of vimentin
was performed by quantitative “Real-Time”-PCR under all treatment conditions simulating diverse tumor regions. Here we demonstrated
that vimentin expression patterns in human malignant glioma cells strongly depend on cellular density, algorithms of drug
delivery and chemo/radio treatment. Substantial differences were recognized between immediate and late therapy effects. Significant
increase in vimentin expression levels was detected particularly in low-density cell cultures under durable treatment with
constant concentration levels of temezolomide. Simulation of variable intratumoral regional conditions (central intratumoral
regions vs. disseminated malignant cells in peripheral regions) demonstrated differential response of vimentin expression
in malignant glioma cell cultures treated under clinically relevant conditions. Slight ebbing of expression levels as late
effects of the treatment in confluent cultures may correspond to necrotic processes clinically observed in central intratumoral
regions. Contrary, in disseminated malignant cells of peripheral regions therapy resulted in vimentin-inducing effects. This
is in agreement with the clinical observations of an increased aggressiveness and malignancy grade of post-operatively chemo/radio-treated
malignant gliomas. 相似文献
74.
Maureen P. Martin Anita Harding Robert Chadwick Mel Kronick Michael Cullen Ling Lin Emmanuel Mignot M. Carrington 《Immunogenetics》1997,47(2):131-138
The human genome contains a large number of interspersed microsatellite repeats which exhibit a high degree of polymorphism
and are inherited in a Mendelian fashion, making them extremely useful genetic markers. Several microsatellites have been
described in the HLA region, but allele nomenclature, a set of broadly distributed controls, and typing methods have not been standardized, which
has resulted in discrepant microsatellite data between laboratories. In this report we present a detailed protocol for genotyping
microsatellites using a semi-automated fluorescence-based method. Twelve microsatellites within or near the major histocompatibility
complex (MHC) were typed in the 10th International Histocompatibility Workshop homozygous typing cell lines (HTCs) and alleles
were designated based on size. All loci were sequenced in two HTCs providing some information on the level of complexity of
the repeat sequence. A comparison of allele size obtained by genotyping versus that obtained by direct sequencing showed minor
discrepancies in some cases, but these were not unexpected given the technical differences in the methodologies. Fluorescence-based
typing of microsatellites in the MHC described herein is highly efficient, accurate, and reproducible, and will allow comparison
of results between laboratories.
Received: 10 May 1997 / Revised: 1 August 1997 相似文献
75.
To investigate the effect of human pyruvate carboxylase (hPC) on lactate formation in Chinese hamster ovary (CHO) cell lines,
FLAG-tagged hPC was introduced into a dihydrofolate-deficient CHO cell line (DG44). Three clones expressing high levels of
hPC, determined by Western blotting using an anti-FLAG monoclonal antibody, and a control cell line were established. Immunocytochemistry
revealed that a substantial amount of expressed hPC protein was localized in the mitochondria of the cells. hPC expression
did not impair cell proliferation. Rather, it improved cell viability at the end of adherent batch cultures with the serum-containing
medium probably because of reduced lactate formation. Compared with control cells, specific lactate production rate of the
three clones was decreased by 21–39%, which was because of a decreased specific glucose uptake rate and yield of lactate from
glucose. Reduced lactate formation by hPC expression was also observed in suspension fed-batch cultures using a serum-free
medium. Taken together, these results demonstrate that through the expression of the hPC enzyme, lactate formation in CHO
cell culture can be efficiently reduced. 相似文献
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79.
M. Pickford 《Human Evolution》1991,6(3):263-268
Being based solely on neontological data, all «unique parent» evolutionary hypotheses, of which «Mitochondrial Eve» is one, fall into the category ofscala naturae. Mathematical treatment of neontological data bases, using cladistic approaches does not confer the status of scientific hypotheses onto such scenarios. Apart from these fundamental problems, such hypotheses are flawed on a number of other bases, including the fact that there is a proportion of parental contribution to mitochondrial lineages, despite widely publicised statements that mithocondrial DNA in mammals is «strictly» maternally inherited. Other weaknesses of «unique mother» hypotheses on that their proponents endeavour to describe the evolution of diploid organisms on the basis of variability in extant haploid organelles, the evolution of which is delinked from that of the diploid organism. A further difficulty is that it is not possible to reconstruct interspecific relationships on the basis of intraspecific variability. There is a general ignorance among proponents of «unique mother» hypotheses regarding the distribution of biological variability on the surface of the globe, a fact which renders the molecular clock inaccurate, and which upsets the simplistic proposal that molecular diversity equates with time. «Unique mother» scenarios are also invalidated by the presence of shared chromosome and other polymorphisms in african great apes and humans at similar percentages in the different lineages, a fact which indicates that these evolving populations did not experience «bottlenecks». These and other difficulties effectively refute the «Mitochondrial Eve» hypothesis, which in any case much resembles creationism of a special kind, in which the offspring of a breeding pair are visualised as belonging to a species different from its parents. Such extreme examples of the punctuational mode of evolution are highly likely to be incorrect. 相似文献
80.
Gene mapping by linkage and association analysis 总被引:3,自引:0,他引:3
March RE 《Molecular biotechnology》1999,13(2):113-122
Genetic analysis is used to map genes, including disease loci, to positions within the human genome. Linkage analysis depends
on the co-segregation of a gene (locus) and a phenotype through a pedigree, while association analysis, or linkage disequilibrium
mapping, depends on measuring deviation from the random occurrence of alleles in a haplotype in unrelated individuals or nuclear
families. Complex computer programs may be used in both forms of analysis. In recent years most interest has focused on identifying
genes involved in common, multifactorial diseases. Here I review some current and developing techniques of genetic analysis
and give references to where further information can be obtained. 相似文献