全文获取类型
收费全文 | 5619篇 |
免费 | 538篇 |
出版年
2023年 | 31篇 |
2022年 | 64篇 |
2021年 | 174篇 |
2020年 | 83篇 |
2019年 | 99篇 |
2018年 | 114篇 |
2017年 | 130篇 |
2016年 | 175篇 |
2015年 | 325篇 |
2014年 | 388篇 |
2013年 | 379篇 |
2012年 | 498篇 |
2011年 | 449篇 |
2010年 | 288篇 |
2009年 | 239篇 |
2008年 | 364篇 |
2007年 | 322篇 |
2006年 | 314篇 |
2005年 | 274篇 |
2004年 | 288篇 |
2003年 | 226篇 |
2002年 | 249篇 |
2001年 | 59篇 |
2000年 | 37篇 |
1999年 | 45篇 |
1998年 | 43篇 |
1997年 | 48篇 |
1996年 | 44篇 |
1995年 | 28篇 |
1994年 | 25篇 |
1993年 | 33篇 |
1992年 | 25篇 |
1991年 | 33篇 |
1990年 | 22篇 |
1989年 | 23篇 |
1988年 | 15篇 |
1987年 | 14篇 |
1986年 | 23篇 |
1985年 | 20篇 |
1984年 | 9篇 |
1983年 | 13篇 |
1982年 | 10篇 |
1981年 | 10篇 |
1980年 | 10篇 |
1979年 | 10篇 |
1978年 | 6篇 |
1976年 | 12篇 |
1973年 | 7篇 |
1972年 | 6篇 |
1970年 | 6篇 |
排序方式: 共有6157条查询结果,搜索用时 640 毫秒
61.
62.
Immunohistochemical identification of tubular segments in percutaneous renal biopsies 总被引:1,自引:0,他引:1
Summary To identify the renal cortical tubular segments involved in tubulo-interstitial disease in formalin-fixed, paraffin-embedded percutaneous kidney biopsies, we developed multiple immunolabeling protocols using segment-specific tubular markers. The present study of biopsies from patients with minimal change or thin basement membrane nephropathy provides a baseline for interpretation of histopathology. Proximal tubules were stained either by the PAS reaction or by the biotinylated Phaseolus vulgaris erythroagglutinin (PHA-E)-streptavidin-gold-silver system (brush borders black). The anti-Tamm-Horsfall (THP) antibody-immunoperoxidase (aminoethylcarbazole, AEC-IPO), and anti-epidermal cytokeratins (ECK) antibodies-immunoalkaline-Fast Blue BB methods marked the distal straight tubules and the cortical collecting system red-brown and blue, respectively. When these immunolabelings were combined, the coapplication of AEC-PO-labeled peanut agglutinin (PNA) or anti-epithelial membrane antigen antibody-AEC-IPO technique (both are markers for distal nephron) visualized the apical membranes of distal convoluted tubules. In the protocol PHA-E + PNA + THP + ECK, the tubular basement membranes were outlined by the anti-laminin antibody-AEC-IPO staining, carried out simultaneously. The protocol PNA + THP + ECK + PAS was found to be a quite appropriate multiple immunolabeling method for the tubules, and is recommended for use as a tool in the study of tubulo-interstitial diseases.Abbreviations PAS
periodic acid-Schiff reaction
- PHAE
Phaseolus vulgaris erythroagglutinin
- PNA
Peanut agglutinin
- EMA
epithelial membrane antigen
- THP
Tamm-Horsfall glycoprotein
- ECK
epidermal cytokeratins
- PO
peroxidase
- Biot-PHA-E
biotinylated PHA-E
- APAAP
complexes of alkaline phosphatase and mouse monoclonal anti-alkaline phosphatase
- SWARI
swine anti-rabbit immunoglobulins
- FCS
fetal calf serum
- TBS
Tris-buffered saline
- AEC
aminoethylcarbazole
- DAB
diaminobenzidine
- FBBB
Fast Blue BB
- IA
immunoalkaline
- GL
glomerulus
- PT
proximal tubule
- DST
distal straight tubule
- DCT
distal convoluted tubule
- CCS
cortical collecting system
- CT
connecting tubule
- CD
collecting duct 相似文献
63.
The gene for the purine salvage enzyme hypoxanthine phosphoribosyltransferase (HPRT) is expressed at a low level in many cells. As is the case with several other “housekeeping genes,” thorough studies of hprt gene regulation have been hampered by the low levels of its mRNA. We have used RNA/RNA hybridization in solution to determine the concentration of hprt-RNA in human cells. The sensitivity and specificity of the method have been validated, and it is shown that hprt-RNA can be accurately determined at a level of a few mRNA molecules per cell. As expected for a housekeeping gene, low and relatively constant hprt-RNA levels (0.3–0.8 pg/μg DNA) were found in primary cultures of normal amnion cells and fibroblasts, EBV-transformed lymphoblastoid cell lines, neuroblastoma, glioblastoma, and melanoma cell cultures. While resting lymphocytes were found to contain very low amounts of hprt-RNA, lymphocytes stimulated with phytohemagglutinin (PHA) showed a 10-fold increase to about 0.8–1.2 pg/μg DNA, which corresponds to 6–10 hprt-RNA molecules per cell. The level started to increase about 20 h after PHA stimulation, 5–10 h before the onset of DNA synthesis, and a steady-state level was reached after 2–3 days in culture. In PHA-stimulated lymphocytes from two brothers with inherited HPRT deficiency (LeschNyhans syndrome), the hprt-RNA level in PHA-stimulated lymphocytes was only about 25% of that in normal subjects. In T-cells selected for HPRT deficiency by growth in 6-thioguanine medium, the levels of hprt-RNA were either normal or very low, which probably reflects the different nature of the mutations involved. These results demonstrate the sensitivity of this method for determinations of low levels of RNA and clearly show induction of hprt-RNA after mitogenic stimulation of human lymphocytes. 相似文献
64.
Erik Steen Hansen Jens Knudsen 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1982,721(4):418-424
Heat production, free fatty acid and glycerol release from white adipose tissue fat pads from obese (ob/ob) mice and their lean littermates are determined. Heat production was significantly lower in obese mice compared to lean mice when expressed on wet weight basis but not when expressed on DNA basis. Noradrenaline significantly increased the heat production in fat pads from both groups of animals. However, the increase in heat production due to noradrenaline addition in fat pads from lean mice was significantly higher than in fat pads from obese mice. The release of free fatty acids and glycerol before incubation with noradrenaline was similar from fat pads from both groups of animals. Addition of noradrenaline to the fat pads increased the release of free fatty acids and glycerol in both groups of animals, but the increase was significantly larger from fat pads from lean mice. In the absence of noradrenaline the free fatty acid/glycerol ratio (mol/mol) in the effluent was 7.9:1 and 4.8:1 for lean mice and obese mice, respectively. In the presence of noradrenaline the ratio decreased to 3:1 for both groups of animals. 相似文献
65.
The breeding performance of captive willow ptarmigan on different diets has been studied. The nutritional factors tested were protein concentration, natural feed supplement and grass meal and flavonoid admixture, and effects on egg numbers, fertility, hatchability, chick weights at hatching and 0–14 days mortality have been recorded. The breeding performance of ptarmigan hen in captivity showed great individual variations. Egg numbers were not statistically different in groups fed the different diets. Hens fed a 15 % crude protein died tended to produce smaller chicks with significantly lower viability than chicks from hens fed a 20 % crude protein diet. Supplement of natural feed tended to increase the number of chicks hatched through a combination of tendency to higher egg numbers and improved fertility. These tendencies were, however, statistically nonsignificant. Inclusion of 34 % grass meal to the diet also tended (non-significantly) to improve fertility and hatchability, while inclusion of flavonoids had no positive effect on reproduction. Eggs from captive hens showed significantly lower fertility, and a tendency to lower hatchability than eggs from wild hens. The former difference was probably caused by the close cage confinements for the captive ptarmigan, while the latter condition probably was due to different start of incubation, most of the eggs from wild hens being started naturally. 相似文献
66.
In secondary leaves from spinach plants pretreated in vermiculite for 24 h with 300 mM NaCl, glycinebetaine accumulated at a rate of circa 0.16 mol 100 g-1 Chl d-1 (2 mol g-1 FW d-1), about three times the rate of control plants. The soluble carbohydrate and free amino acid contents did not increase significantly following salinisation until after 4 d when the relative growth rate also decreased. Leaf proline levels remained very low throughout the experimental period. K+ on a tissue water basis remained constant at 200 mM while Cl- and Na+ levels increased linearly to reach 175 and 100 mM respectively after 5 d of saline treatment. The osmotic pressure of leaf tissue also increased from 300 to 500 mosmol kg-1. These experimental conditions were considered suitable to study glycinebetaine biosynthesis and its induction by salinity in the absence of marked growth inhibition or metabolic disturbance. Radioactive labelled [14C]serine, ethanolamine and choline (all 1 mol, 13.3 MBq in 10 l) were fed to detached secondary leaves via the petiole 24 h after the exposure of plants to salt. The rate of isotope incorporation into water soluble products, lipids and residue was measured over a further 24 h. The major metabolic fate of exogenous [14C]choline and [14C]ethanolamine was incorporation into glycinebetaine while less 14C-label was found in phosphatidyl choline and phosphatidyl ethanolamine. Incorporation rates were identical in control and salinised leaves and were adequate to account for observed values of glycinebetaine accumulation previously reported in spinach. In contrast the labelling of glycinebetaine from [14C]serine was twice as great in salinated plants as in the controls. These results, together with short term labelling experiment with [14C]ethanolamine using leaf slices, were consistent with the formation of glycinebetaine via serine, ethanolamine and its methylated derivatives to choline with some control being exerted at the serine level. However a flux through the phosphorylated intermediates is not excluded.From a consideration of these results and the published data on barley subjected to water stress (Hanson and Scott, 1980 Plant Physiol. 66, 342–348) there appear to be significant differences in the biosynthetic pathways in spinach and barley.Abbreviations BHT
butylated hydroxytoluerte (2,6-di-tert-butyl-4-methylphenol)
- C1
one-carbon fragment
- 1,2DG
diglyceride moiety
- DW
day weight
- MCW
methanol-chloroform-water (12:5:1, by vol.)
- PA
phosphatidic acid
- PC
phosphatidyl choline
- PMME
phosphatidyl monomethylethanolamine
- PDME
phosphatidyl dimethylethanolamine
- PE
phosphatidyl ethanolamine
- PPO
2,5-diphenyloxazole
- POPOP
1,4-bis(5-phenyloxazoyl) benzene 相似文献
67.
The marked effects of ethylene on pea stem growth have been investigated. Low temperature and colchicine, both known microtubule depolymerization agents, reverse the effects of ethylene in straight growth tests. Low temperature (6 C) also profoundly reduces the effects of gas in terms of swelling, hook curvature, and horizontal nutation. Deuterium oxide, an agent capable of rigidifying microtubular structure, mimics the effects of ethylene. Electron microscopy shows that microtubule orientation is strikingly altered by ethylene. These findings indicate that some of the ethylene responses may be due to a stabilizing effect on microtubules in plant cells. 相似文献
68.
Four patients developed massive pulmonary embolism after jejuno-ileal bypass for morbid obesity. All patients were in Greenfield's Class IV and were in shock. Severe hypoxia was evidenced in their blood gases. The patients were managed with digitalis, diuretics, Solu-Medrol (methylprednisolone sodium succinate), oxygen, and heparin therapy. Each patient underwent partial vena cava interruption with Mobin Uddin's umbrella, and all four survived without residual complications. 相似文献
69.
Characteristics of a simple, high-resolution flow cytometer based o a new flow configuration. 下载免费PDF全文
A method is described for the quantitative determination of free and bound solute concentrations in the cytoplasm of intact cells. The method includes (a) introduction of a gelatin gel reference phase (RP) into the cytoplasm; (b) diffusion of dissolved substances between cytoplasm and RP, (c) cell quenching to - 196 degrees C to prevent subsequent solute redistributions, (d) ultra-low temperature microdissection to isolate RP and cytoplasm samples, and (e) analysis of isolates for solute and water content. In normal oocytes of the salamander, Desmognathus ochrophaeus, free or RP Na+ and K+ are 21.0 +/- 1.1 and 128.8 +/- 2.4 mu eq/ml, respectively, and vary stoichiometrically in altered oocytes. Overall cytoplasmic concentrations are 75.2 +/- 2.7 mu eq Na+/ml and 88.6 +/- 1.5 mu eq K+/ml. Cytoplasmic chemical activities are 16.2 mu eq Na+/ml and 99.2 mu eq K+/ml, corresponding to activity coefficients of 0.22 and 1.12, respectively. The results demonstrate unambiguously that (a) oocytes actively transport Na+ and K+, and (b) cytoplasm has important binding properties which differentiate it from an ordinary aqueous solution. These cytoplasmic properties are investigated in the following paper. 相似文献
70.
Steen Pedersen Solveig V. Reeh Jack Parker Robert J. Watson James D. Friesen Niels P. Fiil 《Molecular & general genetics : MGG》1976,144(3):339-343
Summary The presence of EF-Tu, RNA polymerase subunit , and EF-G on the dfus-3 genome and EF-Tu, ribosomal proteins L7/L12, and RNA polymerase subunit on the drif
d
18 genome has been confirmed using a two-dimensional gel electrophoresis technique sensitive to changes in isoelectric point and molecular weight. In this system two EF-Tu gene products could not be resolved. Following infection of ultraviolet light-irradiated Escherichia coli with either dfus-3 or drif
d
18, the EF-Tu gene, tufA, near 65 minutes on the genetic map is expressed as 3–4 copies per EF-G molecule. The EF-Tu gene, tufB, near 79 minutes on the genetic map, is expressed at about one-third of this rate. is expressed as 1 copy per EF-G molecule, as 0.14 per EF-G molecule and L7/L12 as 2.5 per EF-G. These figures compare well with the relative amounts found in exponentially-growing cells, in which the ratio of EF-Tu to EF-G is approximately 5. Almost 90% of the total number of proteins (calculated on a molecular weight basis) which theoretically can be encoded on the drif
d
18 have been identified on the two-dimensional gel. 相似文献