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61.
H. Christopher Wilson Nadine C. Milos 《In vitro cellular & developmental biology. Plant》1987,23(5):323-331
Summary This study investigates the nutritional requirements ofXenopus laevis neural crest cells and melanophores developing in vitro. A comparison is made between the growth and differentiation of cells
in serum-containing medium and a chemically defined, serum-free medium that we have designed. Our chemically defined medium
is more efficient than serum-supplemented medium in promoting proliferation of these cells. Several supplements are required
to enhance culture development. These include insulin, α-melanocyte stimulating hormone, somatotropin, luteotrophic hormone,
linoleic acid, uridine, and putrescine. In addition, collagen and fibronectin provide the most conductive environment tested
for cell migration and adhesion.
This work was supported by establishment and major equipment grants from the Alberta Heritage Foundation for Medical Research
to N. C. M. Nadine C. Milos is a Heritage Medical Research Scholar of the Alberta Heritage Foundation for Medical Research. 相似文献
62.
Summary New cell lines, designated as ML-DmDl≈10, were established from dissociated imaginal discs ofDrosophila melanogaster. The culture medium was prepared by mixing in a 1:1 ratio Cross and Sang’s M3(BF) medium, supplemented with 10% heat inactivated
fetal bovine serum (FBS), with the supernatant of a primary embryonic cell culture made in the M3(BF) medium and supplementing
this mixture with insulin. One cell line was established in the medium containing larval hemolymph instead of the primary
culture supernatan, and another was established in fresh M3(BF) medium supplemented with insulin and FBS. In these mediums,
imaginal disc cells first formed aggregates and cellular vesicles within a few weeks followed by the proliferation of thin-layered
cells around them after about 1 mo. Ten cell lines have so far been established from two kinds of imaginal discs and disc
mixtures. The ploidy of these cell lines was predominantly diploid. Population doubling time was about 50 to 70 h at 3 to
10 mo. after initiation of the culture. When the cell aggregates formed in vitro were implanted in metamorphosing larvae,
they differentiated at high frequency into adult cuticular strutures in the early phase of the primary culture. This differentiation
of aggregates was also observed, though at low frequency, in a culture maintained by dilution-transfer for 6 to 15 mo. in
vitro. 相似文献
63.
Summary Newborn rat adipocyte precursors, isolated from inguinal fat pads of 2 day-old NBR rats proliferate and undergo adipose differentiation
in defined medium in the absence of serum when cultivated on polylysine coated dishes in DME-F12 medium supplemented with
fibronectin, insulin, transferrin and FGF. After 7 days in culture in these conditions, 90% of the cells have undergone differentiation
as measured by the increase of G3PDH specific activity and by the accumulation of triglycerides in their cytoplasm. In contrast,
the cells cultivated in the presence of 10% fetal bovine serum, have a limited ability to differentiate. These results indicate
that newborn rat adipocyte precursors from inguinal fat pads do not require the presence of an undefined adipogenic factor
in order to differentiate in culture. In contrast, proliferation and differentiation are dependent on the presence of insulin
in the culture medium. Moreover, the data presented in this paper show that the rat adipocyte precursor culture represents
a rapid and reproducible system for investigating the processes of adipose tissue development and for studying the negative
and positive regulators of the adipose differentiation in a controlled environment.
This work was supported by grants from the Juvenile Diabetes Foundation, File #185221 and from the National Institutes of
Health 1 PO1 CA37589.
Editor’s Statement This paper extends to primary cultures the serum-free methods previously applied to studies of adipocyte
differentiation in established lines. The observation that serum can block differentiation in this system suggests the existence
of previously unrecognized circulating plasma or platelet factors affecting adipocyte differentiation, and the model developed
provides an assay for the identification of these factors. 相似文献
64.
Different hormonal requirements for androgen-independent growth of normal and tumor epithelial cells from rat prostate 总被引:4,自引:0,他引:4
Wallace L. McKeehan Pamela S. Adams Danna Fast 《In vitro cellular & developmental biology. Plant》1987,23(2):147-152
Summary The proliferation of isolated normal prostate epithelial cells from rat and man is androgen-independent and requires cholera
toxin, insulin, dexamethasone, epidermal growth factor (EGF) and one or more polypeptide factors that are concentrated in
bovine neural tissue. The active agents in the neural tissue extract are heparin-binding polypeptides (prostatropins), the
predominant form of which has a molecular weight of 17400 and an acetylalanine at the aminoterminus. Prostatropins supported
a half-maximal increase in normal prostate epithelial cell number at 50 picomolar. The proliferation of primary and serially-cultured
epithelial cells from androgen-responsive Dunning R3327 rat prostate tumors was also androgen-independent, but exhibited dramatic
alterations in response to hormones that stimulated normal cell proliferation. At low cell density, androgen-independent growth
of isolated tumor-derived epithelial cells was independent on cholera toxin, was stimulated by dexamethasone, required insulin
andeither EGFor prostatropin. The presence of either EGF or prostatropin masked the response to the other factor. In the absence of EGF,
purified prostatropins supported a half-maximal increase in tumor cell number at 7 picomolar. Endogenous production of EGF-like
and prostatropin-like factors or both was suggested by the reduced requirement for EGF and prostatropin at high prostate tumor
cell density. These results suggest that anti-hormonal therapies against prostate tumor growth should be based on intervention
with the activity of insulin (or insulin-like factors) or simultaneous intervention with both EGF and prostatropin (or their
homologues).
This work was supported by NIH grants CA 37589 and HL 33847, and grant 1718 from the Council for Tobacco Research.
Editor’s Statement This paper is the first report of the comparison of the hormone requirements of primary cultures of normal
and tumor prostate epithelial cells from the same system. 相似文献
65.
Young Woo Chun Richard B. Hall Loren C. Stephens 《Plant Cell, Tissue and Organ Culture》1986,5(3):179-185
The in vitro shoot proliferation of Populus alba × P. grandidentata was affected by the medium consistency and shoot density, but not by three sizes of vessels. After 4 weeks of culture, the fresh weight and number of shoots per explant on liquid medium were significantly greater than those on agar-solidified medium. In particular, 3.2 shoots, 7 mm or longer per explant, were produced on liquid medium compared with 1.6 shoots per explant or agar-solidified medium. The fresh weight per explant after 4 weeks of culture on liquid medium and agar-solidified medium were 0.68 and 0.25 g, respectively. Increasing the number of shoots per vessel slowed the growth of the explants as measured by fresh weight and the number of shoots produced. There was little difference in the number of shoots produced between vessels with 1 or 2 shoots per vessel, but there were many fewer shoots produced when 3 shoots were placed in each vessel.Journal Paper No. J-11977 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project 2210. 相似文献
66.
Annie Giraud Simone Bouchilloux 《Biochemical and biophysical research communications》1983,111(2):353-359
The synthesis of glycosaminoglycans (GAGs) was investigated in porcine thyroid cells under the influence or not of thyrotropin. After labelling with [3H] glucosamine and [35S] , enriched GAG-fractions prepared from culture media, cells, and eventually substrate adhering materials, were analyzed by cellulose acetate electrophoresis combined with specific degradations. They comprised heparan sulfate and hyaluronic acid together with an unknown sulfated component labile to endo-β-galactosidase. Whereas global labellings of newly made GAGs were not significantly modified by thyrotropin, we reproducibly observed with the hormone a substantial increase in the proportion of hyaluronic acid [3H] label and, when cells organized into follicles, of the proportion of cell-associated [3H] GAGs. This system thus offers an interesting model to study how the responsiveness to an hormone and the reorganization that follows might implicate specific glycoconjugates. 相似文献
67.
Analysis of the biological effects of specific DNA alkylations by simple alkylating agents is complicated by the variety of sites involved. It is, therefore, of value to be able to incorporate into cellular DNA nucleosides alkylated in a single position, e.g., O6-methyldeoxyguanosine. Such cellular incorporation is particularly difficult to achieve because this nucleoside is rapidly demethylated by adenosine deaminase. We have attempted to achieve such incorporation into the DNA of V79 cells by using coformycin, an inhibitor of adenosine deaminase, and by forcing the cells to depend on exogenous purines by the use of medium containing aminopterin. The DNA of V79 cells exposed to O6-methyl-[8-3H]deoxyguanosine (2.4 μM, sp. act. 14 500 Ci/mole) showed an incorporation level of 4 × 10−8 nucleotides. When 1000-fold higher concentrations were employed (3–15 mM, sp. act. 1.6 Ci/mole), significant cytotoxicity and inhibition of DNA synthesis was observed. However, because it was not economically feasible to administer high specific activity O6-methyldeoxyguanosine to the cells at these concentrations, we could not determine the amount of labeled nucleoside incorporated into DNA. Examination of the frequency of 6-thioguanine-resistant cells in these treated populations showed no significant increase above the background level. Comparison of the cytotoxic effect of O6-methyldeoxyguanosine with deoxyadenosine showed that the toxicity induced by O6-methyldeoxyguanosine could have resulted from mimicry of deoxyadenosine, rather than by incorporation of the alkylated nucleoside itself. 相似文献
68.
SYNOPSIS. Media have been developed for axenic cultivation of 10 strains belonging to 7 species of small marine ciliates. A medium containing cerophyl extract, proteose peptone, trypticase, yeast nucleic acid, biotin, calcium pantothenate folic acid, nicotinamide, pyridoxal HCl, riboflavin, thiamine HCl, and DL-thioctic acid in sea water supports the growth of Uronema nigricans strains Pc and 34/2, Parauronema virginiatum strains 2/1 and 19/1, Miamiensis avidus strains Ma and 19/3, Miamiensis sp. strain 1/1, a strain of "G" ciliate, and strains 33/8 and 34/7 of 2 unidentified species. By substituting a mixture of asolecithin, cephalin, and Tween 80 for cerophyl in the medium, luxurious growth of all except the strains of the 2 unidentified species can be obtained. A defined medium consisting of 18 amino acids, 5 purine derivatives, 8 vitamins, asolecithin, cephalin, and Tween 80 in synthetic sea water also has been developed for 6 of the strains: M. avidus Ma and 19/3, Miamiensis sp. 1/1, P. virginiatum 2/1 and 19/1, and U. nigricans Pc. In general the ciliates grow best at pH 7.2 in the dark at 27 C in media containing sea water of density = 1.015. Under these conditions maximum populations are reached in 4–5 days and range from several hundred thousand to 3 or 4 × 106 depending upon the strain. Electronmicroscopic observations for the presence of endosymbiotes gave negative results. 相似文献
69.
N. Hammatt B. Jones M. R. Davey 《In vitro cellular & developmental biology. Plant》1989,25(7):669-672
Summary Plants have been regenerated from nodular, green callus derived from cotyledon, petiole and leaf lamina explants ofG. argyrea, a perennial relative of the soybean (G. max). The degree of response obtained was governed primarily by the genotype used, accession G1626 proving the most responsive.
Shoots were also recovered from about 6.0% of cotyledon protoplasts of this genotype. The implications of these results are
discussed in relation to genetic manipulations using this species. 相似文献
70.
Masahiro Miyazaki Yasunori Suzuki Munehiro Oda Akira Kawai Liyan Bai Jiro Sato 《In vitro cellular & developmental biology. Plant》1989,25(9):839-848
Summary For serum-free primary culture of adult rat hepatocytes, a synthetic medium DM-160 and rat-tail collagen were selected for
the basal medium and for the culture substratum, respectively. Barbiturates, such as phenobarbital and 1-ethyl-5-isobutylbarbiturate,
efficiently supported survival of hepatocytes and maintained their morphologic features at lower concentrations under the
serum-free conditions than under the serum-supplemented conditions. However, the hepatocyte survival rates under the serum-free
conditions were lower than those under the serum-supplemented conditions in the presence or absence of barbiturates. Supplementation
of the basal medium with a combination of five groups of factors (5Fs), such as eight amino acids (Ala, Arg, Gly, Ile, Met,
Phe, Pro, and Trp), two unsaturated fatty acids (linoleate and oleate), a protease inhibitor (aprotinin), three vitamins (A,
C, and E), and five trace elements (Mn, Fe, Cu, Zn, and Se), improved the hepatocyte survival under the serum-free conditions
in the presence or absence of barbiturates. In other words, the serum could be completely substituted by the 5Fs. Hepatocyte
cultures maintained in the 5Fs-suppelemented basal medium showed excellent induction of tyrosine aminotransferase activity
in response to dexamethasone in the presence or absence of barbiturates. The efficiency of the 5Fs-supplemented basal medium
for maintaining hepatocytes was not inferior to those of other media in common use with hepatocytes, such as Williams' medium
E and Waymouth's medium MB-752/1. In conclusion, maintenance of functional hepatocytes in serum-free primary culture could
be improved by use of the new medium preparation (the 5Fs-supplemented DM-160) in the presence of barbiturates.
This work was supported by a grant no. 61771923 from the Ministry of Education, Science and Culture of Japan. 相似文献