全文获取类型
收费全文 | 42546篇 |
免费 | 2232篇 |
国内免费 | 1442篇 |
出版年
2023年 | 541篇 |
2022年 | 837篇 |
2021年 | 1095篇 |
2020年 | 1090篇 |
2019年 | 1496篇 |
2018年 | 1520篇 |
2017年 | 914篇 |
2016年 | 1051篇 |
2015年 | 1260篇 |
2014年 | 2550篇 |
2013年 | 3175篇 |
2012年 | 1789篇 |
2011年 | 2576篇 |
2010年 | 1842篇 |
2009年 | 2023篇 |
2008年 | 2211篇 |
2007年 | 2238篇 |
2006年 | 1986篇 |
2005年 | 1767篇 |
2004年 | 1550篇 |
2003年 | 1335篇 |
2002年 | 1211篇 |
2001年 | 759篇 |
2000年 | 694篇 |
1999年 | 705篇 |
1998年 | 699篇 |
1997年 | 580篇 |
1996年 | 534篇 |
1995年 | 510篇 |
1994年 | 473篇 |
1993年 | 414篇 |
1992年 | 354篇 |
1991年 | 323篇 |
1990年 | 274篇 |
1989年 | 234篇 |
1988年 | 206篇 |
1987年 | 196篇 |
1986年 | 171篇 |
1985年 | 273篇 |
1984年 | 465篇 |
1983年 | 387篇 |
1982年 | 365篇 |
1981年 | 278篇 |
1980年 | 250篇 |
1979年 | 202篇 |
1978年 | 151篇 |
1977年 | 147篇 |
1976年 | 127篇 |
1975年 | 102篇 |
1973年 | 109篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
181.
A soluble Lewis blood-group gene associated -3/4-L-fucosyltransferase has been purified from human milk by a series of steps involving hydrophobic chromatography on Phenyl Sepharose 4B, ion exchange chromatography on CM-Sephadex C-50, affinity chromatography on GDP-hexanolamine Sepharose 4B and gel filtration on Sephacryl S-200. The first step separated -3-L-fucosyltransferase activity directed towardsN-acetylglucosamine in Type 2 (Gal1-4GlcNAc-R) acceptors from an -3/4-fucosyltransferase fraction acting on both Type 1 (Gal1-3GlcNAc-R) and Type 2 acceptors. Further purification of this latter fraction on CM-Sephadex and GDP-hexanolamine Sepharose gave a single peak of fucosyltransferase activity that catalysed the addition of fucose toN-acetylglucosamine in both Type 1 and Type 2 acceptors and to theO-3 position of glucose in lactose-based oligosaccharides. The enzyme preparation at this stage resembled previously described -3/4-fucosyltransferase preparations purified from human milk. However, gel filtration of this preparation on Sephacryl S-200 or Sephadex G-150 separated further amounts of -3-fucosyltransferase activity acting solely on Type 2 acceptors and left a residual -3/4-fucosyltransferase that retained strong -4 activity with the Type 1 acceptor, lacto-N-biose 1, and -3 activity with 2-fucosyllactose, but had relatively little -3 activity withN-acetyllactosamine and virtually no capacity to transfer fucose to glycoproteins withN-linked oligosaccharide chains having unsubstituted terminal Type 2 structures. 相似文献
182.
Chronic treatment of hamsters with estradiol for several months has previously been shown to decrease the specific content
of cytochrome P450 in the kidney, a target of hormonal carcinogenesis, but not in liver. The reason for this decrease in metabolic
enzyme activity is unknown and has been examined in this investigation. We now report that the decrease in specific content
of renal cytochrome P450 by 73% in response to estradiol was not affected by co-treatment with tamoxifen for 1 month. The
subcutaneous infusion of 250 μg/day estradiol for 7 days lowered renal cytochrome P450 by 71% from control values and was
therefore used for further mechanistic studies. This treatment decreased renal activities of estradiol 2- or 4-hydroxylase
by 77 to 80%, of 7-ethoxycoumarin-O-deethylase by 66% of control values, respectively, and completely eliminated aryl hydrocarbon
hydroxylase activities, whereas liver enzymes remained unaffected. After 7 days of infusion of estradiol, fluorescent products
of lipid peroxidation were more than doubled in hamster kidney but remained unchanged in liver. The possibility of enzyme
destruction by binding of estradiol 2,3-quinone to metabolizing enzymes was investigatedin vitro. In the presence of 2-hydroxyestradiol, cumene hydroperoxide, and microsomes, conditions known to favor the oxidation of
the steroid to quinone, the binding of catechol estrogen metabolite to microsomal protein increased 60 fold over control values
in the absence of cofactor. Purified rat liver cytochrome P450c also oxidized 2-hydroxyestradiol to 2,3-estradiol quinone.
The rate of oxidation was linear for the first 2–3 min, but thereafter decreased with time. Under these incubation conditions,
irreversible binding of catechol estrogen metabolite to cytochrome P450c increased for the first 2–3 min and then remained
at this plateau level. It was concluded that enzyme destruction by a reactive estrogen metabolite or by lipid peroxides may
be a major reason for the organ-specific decrease in cytochrome P450 enzymes in kidneys of estrogen-treated hamsters. 相似文献
183.
The 130 kDa atrial natriuretic factor receptor (ANF-R1) purified from bovine adrenal zona glomerulosa is phosphorylated in vitro by serine/threonine protein kinases such as cAMP-, cGMP-dependent and protein kinase C. This phosphorylation is independent of the presence of ANF (99–126) and there is no detectable intrinsic kinase activity associated with the ANF-R1 receptor or with its activated form. In bovine adrenal zona glomerulosa cells, TPA (phorbol ester) induces a marked inhibition of the ANF-stimulated cGMP accumulation as well as of the membrane ANF-sensitive guanylate cyclase catalytic activity without any change in the binding capacity or affinity for 125I-ANF. However, we have demonstrated a significant 32P incorporation in the ANF-R1 receptor of the TPA-treated cells. The effect of TPA on the zona glomerulosa ANF-R1 receptors was abolished by calphostin C, a specific protein kinase C inhibitor. Altered ANF actions due to blunted response of guanylate cyclase to ANF could be a consequence of the ANF receptor phosphorylation by excessive activity of protein kinase C and might be involved in the pathogenesis of hypertension.Abbreviations ANF
Atrial Natriuretic Factor
- ANF-R1
Atrial Natriuretic Factor Receptor, subtype 1
- ATP
Adenosine Triphosphate
- CaCl2
Calcium Chloride
- cAMP
Adenosine cyclic 3,5-Monophosphate acid
- cGMP
Guanosine cyclic 35-Monophosphate acid
- EDC
1-Ethyl-3-[3-Dimethylaminopropyl] Carbodiimide
- EDTA
Ethylenediaminetetraacetic Acid
- GTP
Guanosine Triphosphate
- IBMX
3-isobutyl-1-methylxanthine
- kDa
Kilodaltons
- MgCl2
Magnesium Chloride
- MgAC
Magnesium Acetate
- NaCl
Sodium Chloride
- PAGE
Polyacrylamide Gel Electrophoresis
- PKA
cAMP-dependent protein kinase
- PKG
cGMP-dependent Protein Kinase
- PKC
Calcium/Phospholipid-dependent Protein Kinase
- RIA
Radioimmunoassay
- SDS
Sodium Dodecyl Sulfate
- SHR
Spontaneously Hypertensive Rat
- Tris HCl
Tris (Hydroxymethyl) aminomethane Hydrochloride
- TPA
12-O-Tetradecanoyl-Phorbol-13-Acetate 相似文献
184.
The prolactin receptor is a membrane protein mainly involved in the development of the mammary gland and in lactation in mammals. We used specific cDNA constructs and the insect/baculovirus expression system and produced independently and in large amounts several recombinant forms of the rabbit mammary gland prolactin receptor: the full-length receptor (L1, L2), a truncated membrane form (S), a secretable form of the extracellular domain (E) and two forms of the intracellular domain (I1, I2). Of these forms, the L1 and L2 are associated with the membrane fraction, the E is predominantly secreted into the medium and the I1 and I2 are expressed as soluble proteins and surprisingly, a great portion accumulates in the culture medium. The molecular mass (94 kDa) of the expressed full-length receptor corresponds to the translation product of the entire cDNA coding region. The receptor biochemically identified in the rabbit mammary gland is however much shorter. Thus, in the mammary gland, the receptor presumably undergoes post-translational modifications. The receptor forms L1, L2 and S bind prolactin with specificity and affinity similar to those reported for the native receptor. They also interact with two monoclonal antibodies, M110 and A917, specific for the native conformation of the hormone-binding site. The I1 and I2 forms do not bind prolactin, whereas the E form does. Thus, the hormone binding site is located in the extracellular domain which can function autonomously as a PRL-binding soluble protein. However, the E form binds prolactin with a higher affinity than the native receptor and it does not bind one of the two antireceptor monoclonal antibodies, known to be hormone binding-site specific. Thus, the conformation of the native receptor and that of the E form differ. 相似文献
185.
J. L. Tholozan J. P. Touzel E. Samain J. P. Grivet G. Prensier G. Albagnac 《Archives of microbiology》1992,157(3):249-257
Strain X4 was isolated several years ago from an anaerobic mesophilic plant treating vegetable cannery waste waters. It was the first example of propionic fermentation from ethanol. Morphologic and physiologic characterizations of the strain are presented here. This strain is described as type strain of a new species, Clostridium neopropionicum sp. nov. Whole cells of strain X4 ferment [1-13C]ethanol and CO2 to [2-13C]propionate, [1-13C]acetate and [2-13C]propanol, suggesting the absence of a randomizing pathway during the propionate formation. Enzymes involved in this fermentation were assayed in cell-free extracts of cells grown with ethanol as sole substrate. Alcohol dehydrogenase, aldehyde dehydrogenase, phosphate acetyl transferase, acetate kinase, pyruvate synthase, lactate dehydrogenases, and the enzymes of the acrylate pathway were detected at activities sufficient to be involved in ethanol fermentation. The same pathway may be used for the degradation of lactate or acrylate to acetate. 相似文献
186.
Immunogold labelling of the cytoplasmic estradiol receptor in resting porcine endometrium 总被引:2,自引:0,他引:2
Summary Serial sections of resting porcine endometrium were analyzed with the monoclonal antibody 13H2 using goat antimouse IgG/5 nm gold as secondary reagent or with either polyclonal antibodies from goat #402 or the rat monoclonal antibody H222, both in combination with protein G/12 nm gold. A modestly higher labelling of nuclei than of cytoplasm was seen only with the monoclonal antibody H222. Polyclonal #402 and monoclonal 13H2 showed fewer attachments over nuclear than over cytoplasmic areas. The highest densities of attachment and of predominantly cytoplasmic labelling were obtained with the monoclonal antibody 13H2. The results confirm the earlier assumption of a restricted accessiblity of estradiol receptor in the cytoplasm of resting cells for immunoreagents. 相似文献
187.
The activity of -l--aminoadipyl)-l-cysteinyl-d-valine catalysis. Addition of l-cysteine to fermentation media increased -lactam production in both organisms and alleviated the negative carbon source regulation by glycerol in S. clavuligerus. 相似文献
188.
The mechanism responsible for the initial steps in the anaerobic degradation of trans-cinnamate and -phenylalkane carboxylates by the purple non-sulphur photosynthetic bacterium Rhodopseudomonas palustris was investigated. Phenylacetate did not support growth and there was a marked CO2 dependence for growth on acids with greater side-chain lengths. Here, CO2 was presumably acting as a redox sink for the disposal of excess reducing equivalents. Growth on benzoate did not require the addition of exogenous CO2. Aromatic acids with an odd number of side-chain carbon atoms (3-phenylpropionate, 5-phenylvalerate, 7-phenylheptanoate) gave greater apparent molar growth yields than those with an even number of side-chain carbon atoms (4-phenylbutyrate, 6-phenylhexanoate, 8-phenyloctanoate). HPLC analysis revealed that phenylacetate accumulated and persisted in the culture medium during growth on these latter compounds. Cinnamate and benzoate transiently accumulated in the culture medium during growth on 3-phenylpropionate, and benzoate alone accumulated transiently during the course of trans-cinnamate degradation. The transient accumulation of 4-phenyl-2-butenoic acid occurred during growth on 4-phenylbutyrate, and phenylacetate accumulated to a 1:1 molar stoichiometry with the initial 4-phenylbutyrate concentration. It is proposed that the initial steps in the anaerobic degradation of trans-cinnamate and the group of acids from 3-phenylpropionate to 8-phenyloctanoate involves -oxidation of the side-chain.Abbreviation 3-PP
3-phenylpropionic acid
- 4-PB
4-phenylbutyric acid
- 5-PV
5-phenylvaleric acid
- 6-PH
6-phenylhexanoic acid
- 7-PH
7-phenylheptanoic acid
- 8-PO
8-phenyloctanoic acid
- 4-P2B
4-phenyl-2-butenoic acid
- GC/MS
Gas chromatography/Mass spectrometry
- HPLC
High-pressure liquid chromatography 相似文献
189.
Comamonas acidovorans NBA-10 was previously shown to degrade 4-nitrobenzoate via 4-hydroxylaminobenzoate and 3,4-dihydroxybenzoate. Washed cells, grown on a mixture of 4-nitrobenzoate and ethanol, stoichiometrically produced ammonium and 3,4-dihydroxybenzoate from 4-nitrobenzoate under anaerobic conditions provided ethanol was present. In cell extracts 4-hydroxylaminobenzoate was degraded to ammonium and 3,4-dihydroxybenzoate, but this activity was lost upon dialysis. No requirement for a cofactor was found, but rather reduced incubation conditions were necessary to restore enzyme activity. The 4-hydroxylamino-degrading enzyme was purified and the role of this novel type of enzyme in the degradation of nitroaromatic compounds is discussed.Abbreviation 4-ABA
4-aminobenzoate
- 4-NBA
4-nitrobenzoate
- 4-HABA
4-hydroxylaminobenzoate
- 3,4-diHBA
3,4-dihydroxybenzoate 相似文献
190.
Koji Adachi Paul Belser Hans Bender Derui Li Ulrich Rodeck Etty N. Benveniste David Woo Wolff H. Schmiegel Dorothee Herlyn 《Cancer immunology, immunotherapy : CII》1992,34(6):370-376
Summary Recombinant tumor necrosis factor (rTNF; optimal dose 1000 U/ml) significantly increased the density of epidermal growth factor receptor (EGF-R) in three of four glioma cell lines in culture as determined by binding analysis of anti-EGF-R monoclonal antibody (mAb) 425. Since enhancement of EGF-R expression by rTNF- was inhibited when cells were treated with the protein synthesis inhibitor cycloheximide, the effects of rTNF may be protein-synthesis-dependent. The dose of rTNF that was optimal for up-regulation of EGF-R on glioma cells did not inhibit the growth of these cells.125I-labeled mAb 425 lysed glioma cells in culture following its internalization into the cells. After glioma cells had been treated with rTNF, the growth-inhibitory effects of the mAb were significantly enhanced, probably a reflection of the increase in EGF-R density on the tumor cell surfaces. The rTNF effects were specific to the EGF-R and did not affect unrelated glioma-associated antigens. In our previous clinical trials,125I-labeled mAb 425 showed immunotherapeutic effects in glioma patients. The present study provides the basis for considerations of combined immunotherapy of glioma patients with125I-labeled mAb 425 and rTNF. 相似文献