全文获取类型
收费全文 | 18600篇 |
免费 | 926篇 |
国内免费 | 27篇 |
专业分类
19553篇 |
出版年
2024年 | 31篇 |
2023年 | 164篇 |
2022年 | 376篇 |
2021年 | 706篇 |
2020年 | 414篇 |
2019年 | 446篇 |
2018年 | 647篇 |
2017年 | 604篇 |
2016年 | 791篇 |
2015年 | 897篇 |
2014年 | 1134篇 |
2013年 | 1562篇 |
2012年 | 1648篇 |
2011年 | 1445篇 |
2010年 | 837篇 |
2009年 | 731篇 |
2008年 | 850篇 |
2007年 | 824篇 |
2006年 | 688篇 |
2005年 | 621篇 |
2004年 | 502篇 |
2003年 | 419篇 |
2002年 | 373篇 |
2001年 | 308篇 |
2000年 | 280篇 |
1999年 | 254篇 |
1998年 | 101篇 |
1997年 | 76篇 |
1996年 | 83篇 |
1995年 | 77篇 |
1994年 | 57篇 |
1993年 | 56篇 |
1992年 | 152篇 |
1991年 | 143篇 |
1990年 | 113篇 |
1989年 | 85篇 |
1988年 | 126篇 |
1987年 | 97篇 |
1986年 | 80篇 |
1985年 | 81篇 |
1984年 | 81篇 |
1983年 | 46篇 |
1982年 | 40篇 |
1981年 | 47篇 |
1980年 | 43篇 |
1979年 | 56篇 |
1978年 | 35篇 |
1977年 | 43篇 |
1976年 | 31篇 |
1974年 | 29篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
81.
Mini-mu derivatives carrying plasmid replicons can be used to clone genesin vivo. This method was adopted to generate phasmid clones which were later screened for their ability of restore nitrofurantoin
sensitivity of a nitrofuran-resistant host by eliciting nitroreductase activity. One phasmid-derived clone (pAJ101) resulted
in considerable increase in nitroreductase activity when introduced into a nitrofurantoin-resistant mutant ofEscherichia coli with reduced nitroreductase activity. Subsequently, a 1.8 kb fragment obtained from pAJ101 by partial digestion with 5au3A,
was subcloned into pUC18 to yield pAJ102. The nitroreductase activity attributable to pAJ102 was capable of reducing both
nitrofurantoin and nitrofurazone. The polypeptides encoded by pAJ102 were identified by the minicell method. A large, well-defined
band corresponding to 37 kDa and a smaller, less-defined band corresponding to 35 kDa were detected. Tnl000 mutagenesis was
used to delineate the coding segment of the 1.8 kb insert of pAJ102. A 0.8 kb stretch of DNA was shown to be part of the nitroreductase
gene. The gene was mapped at 19 min on theEscherichia coli linkage map. 相似文献
82.
Regulation of phosphorylation of the c-erbB-2/HER2 gene product by a monoclonal antibody and serum growth factor(s) in human mammary carcinoma cells. 总被引:6,自引:0,他引:6
下载免费PDF全文
![点击此处可从《Molecular and cellular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Monoclonal antibody (MAb) 4D5 was used to analyze the phosphorylation of p185HER2, the gene product of c-erbB-2/HER2, in SK-BR-3 cells. Culture in the continuous presence of 4D5 reduced the in vivo steady-state levels of p185HER2 phosphorylation by 80% in a dose-dependent manner, suggesting that MAb 4D5 may have interfered with the activation of phosphorylation of p185HER2. The observed MAb-mediated reduction of p185HER2 phosphorylation could not be completely accounted for by down-regulation. When cultures were grown under serum-free conditions, the steady-state levels of p185HER2 phosphorylation were reduced by 56%, and addition of 4D5 further inhibited phosphorylation to 20% of steady-state levels. With continuous exposure to increasing concentrations of newborn calf serum in these cultures, there was a linear increase in tyrosine-specific phosphorylation of p185HER2, reaching a 5.4-fold increase with 10% newborn calf serum. Phosphorylation of p185HER2 in the presence of newborn calf serum was not attributable to stimulation of the epidermal growth factor receptor by epidermal growth factor or by transforming growth factor-alpha. Extension of these observations to two other mammary carcinoma cell lines. MDA-MB-453 and BT-474, also demonstrated a significant capacity of serum to induce p185HER2 phosphorylation. The demonstration of antibody-mediated partial inhibition of phosphorylation under serum-free conditions suggests that mammary carcinoma cells may also produce and secrete a factor or factors which may activate p185HER2. Our observation that growth-inhibitory MAb 4D5 is able to reduce the phosphorylation of p185HER2 by newborn calf serum and by a cellular-derived factor(s) suggests the existence of a growth factor(s) which uses phosphorylation of p185HER2 as a signal transduction pathway to regulate cell proliferation. 相似文献
83.
Sujata Misra Kshudiram Naskar Dwijen Sarkar Dilip Kumar Ghosh 《Molecular and cellular biochemistry》1991,102(1):13-18
Summary
Leishmania donovani, the etiological agent for the disease visceral leishmaniasis, attach themselves to the macrophages for initiation of the disease. The attachment process has been found to be regulated by Ca2+ ions. Verapamil, a Ca2+-channel blocker inhibits Leishmania-macrophage attachment. The inhibitory effect is increased with time. Nifedipine, another Ca2+-channel blocker exhibits the same effect. The attachment process is stimulated by Ca2+-ionophore alone. The inhibitory effects of the calcium channel blockers are reversed by the ionophore. 相似文献
84.
The cytopathologic features of two cases of peritoneal endometriosis (secondary to ruptured ovarian endometrial cysts) are described. Both patients presented with abdominal distension and tenderness and were clinically thought to have an abdominal tumor. Preoperative cytologic examination of peritoneal fluids gave a diagnosis of endometriosis in both cases. The endometrial tissue was present in the smears as honeycombs, syncytia and tight clusters of both epithelial and stromal cells. Subsequent surgery confirmed the cytodiagnosis in both cases. These cases emphasize the need to include endometriosis in the differential diagnosis of peritoneal effusions, especially in women. 相似文献
85.
The regulatory mechanism for the hormonal control of prolactin (PRL) and growth hormone (GH) gene expression in rat pituitary gland during gestation and lactation is verified in this study. The level of PRL-specific mRNA (mRNAPRL) sequences in the pituitary gland is elevated in the later part of gestation and more prominently so in lactation. In contrast the expression of GH gene is inhibited in the same tissue during gestation and lactation resulting in the dramatic decrease in the level of GH-specific mRNA (mRNAGH) sequences. We now demonstrate the influence of a tissue-specific altered DNA methylation pattern on the temporal modulation of expression of PRL and GH genes in the pituitary gland during alternate physiological states. An altered methylation pattern of specific "-C-" residues only in the coding region of PRL and GH gene can be detected concurrently with the altered level of expression of these genes in the pituitary gland during gestation and lactation. These results also demonstrate the dynamic state of methylation of specific -C- residues during the transition of these two genes from one state of expression to another in the same tissue. A correlation between site-specific DNA methylation and tissue-specific expression of PRL and GH gene in pituitary gland is reported. Thus a role of DNA methylation in the hormonal control of PRL and GH gene expression in physiological states such as pregnancy and lactation is proposed. 相似文献
86.
C B Siegall T T Hla A Kumar 《Biochemical and biophysical research communications》1988,154(3):1010-1017
Functional reconstitution of U1 small nuclear ribonucleoprotein particle (U1 snRNP) was performed using in vitro transcribed U1 snRNA. Hela cell nuclear extract was depleted of its constituent snRNPs by centrifugation at 100,000 X g. The supernatant was devoid of snRNAs and lacked cleavage activity in splicing reactions using in vitro transcribed beta-globin pre-mRNA as substrate. The resulting pellet which contained the snRNAs, retained 5' splice site cleavage activity in a similar splicing reaction. Supplementation of the inactive supernatant fraction with in vitro transcribed U1 snRNA, partially restored 5' splice site cleavage activity thereby demonstrating the specific requirement of U1 snRNP in the initial stage of pre-mRNA splicing. 相似文献
87.
The high molecular weight aminoacyl-tRNA synthetase complex (the 24S complex) was isolated from rat liver by ultracentrifugation. The lysyl-tRNA synthetase (E.C. 6.1.1.6) was selectively dissociated by hydrophobic interaction chromatography on 1,6 diaminohexyl agarose followed by hydroxylapatite chromatography and DEAE chromatography. The lysyl-tRNA synthetase dissociated from the 24S synthetase complex was purified approximately to 2700 fold with 14% yield. 相似文献
88.
Asymmetry of lysophosphatidylcholine/cholesterol vesicles is sensitive to cholesterol modulation 总被引:4,自引:0,他引:4
Sonication of lysophosphatidylcholine (lysoPC; 20 mumol/mL) and cholesterol (chol) in aqueous medium produces lamellar structures over a wide range of concentrations. From 25 to 47 mol % cholesterol, electron microscopy (EM) after negative staining showed extended stacklike lamellae about 40 A thick. From 50 to 60 mol % chol, freeze-fracture EM showed homogeneous populations of small unilamellar vesicles averaging 260-310 A in diameter. Phosphorus-31 nuclear magnetic resonance was used to characterize the stacklike lamellae and to measure the distribution of the lysophospholipid between the outer and inner leaflet of the vesicles as a function of sterol concentration. We found that in lysoPC/chol dispersions containing less than equimolar amounts of cholesterol (25-47 mol %), the entire phosphorus signal (40.5 ppm) was shifted downfield by 10.5 ppm upon addition of Pr3+ (2.4 mM), consistent with the stacklike lamellar structures in which all lysoPC head groups are accessible to the ions. By contrast, addition of Pr3+ to lysoPC/chol vesicles containing equimolar or higher amounts of cholesterol (up to 60 mol %) gave rise to two phosphorus peaks. The more intense downfield signal (51.0 ppm) responsive to paramagnetic ions was assigned to lysoPC located in the outer vesicle leaflet. The upfield signal (40.5 ppm), which was not affected by the ions, was assigned to inside lysoPC. For lysoPC/chol (1:1) vesicles, an outside to inside lysophospholipid ratio (Ro/i) of 6.5 was determined. Essentially the same Ro/i value (6.7) was obtained on lysoPC/chol (1:1) vesicles which after dialysis contained only entrapped Pr3+.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
89.
L C Petersen M Johannessen D Foster A Kumar E Mulvihill 《Biochimica et biophysica acta》1988,952(3):245-254
A one-chain recombinant tissue-type plasminogen activator (EC 2.4.31.-) (tPA) analogue was constructed in which Arg-275 of the activation site was changed to Gly by site-directed mutagenesis. This analogue, tPA-Gly275, was very resistant to plasmin (EC 2.4.21.5) cleavage. It has been used to gain information about the activity of the uncleaved one-chain tPA form, also when plasmin is generated as a result of a plasminogen activation reaction. The amidolytic activity of tPA-Gly275 with less than Glu-Gly-Arg-pNA was investigated and compared to that of one-chain and two-chain wild-type recombinant tPA. A small but significant intrinsic amidolytic activity was observed with the analogue as well as the wild-type one-chain tPA form. However, it was much lower than that of two-chain tPA. Polymerised fibrin enhanced the amidolytic activity of both one-chain tPA forms but not of two-chain tPA. Measurements of the plasminogen activation kinetics in the absence of fibrin revealed that tPA-Gly275 possessed a significant intrinsic activity. However, it was 30-fold lower than that of two-chain tPA. Addition of polymerised fibrin profoundly enhanced the plasminogen activation rate of both tPA-Gly275 and wild-type one- and two-chain tPA to approximately the same maximal level. The results were interpreted to mean that fibrin binding can induce an activated state of the intact tPA one-chain form. 相似文献
90.
Using the overlapping deficiencies Df(3R)GC14 and Df(3R)e
Gp
4 of the 93D region of Drosophila melanogaster, the benzamide (BM)-inducible site in polytene chromsomes was localised to the 93D6-7 region, which had earlier been identified as heat inducible. The normal developmental and BM-induced 93D6-7 puff was found to be dosage compensated since in larvae heterozygotus for a deficiency, with one dose of 93D6-7, the rate of 3H-uridine incorporation in this puff was the same as in the wild type with two doses. Curiously, however, heat shock (37° C) caused regression of the 93D6-7 puff on the normal chromosome in heterozygotes. In agreement with earlier results from our laboratory, the non-inducibility of the single-dose 93D locus by heat shock in the heterozygotes, caused the 87C puff to be nearly half as active as the 87A puff at 37° C. However, in e
Gp
4/GC14 larvae, lacking the 93D6-7 locus on both homologues, the 87C puff was less active than 87A in some heat-shocked larvae but in other larvae 87A and 87C were equally active. Possible reasons for this inter-larval variability are discussed. 相似文献