全文获取类型
收费全文 | 8228篇 |
免费 | 292篇 |
国内免费 | 236篇 |
出版年
2023年 | 117篇 |
2022年 | 77篇 |
2021年 | 149篇 |
2020年 | 169篇 |
2019年 | 198篇 |
2018年 | 198篇 |
2017年 | 142篇 |
2016年 | 166篇 |
2015年 | 172篇 |
2014年 | 195篇 |
2013年 | 572篇 |
2012年 | 147篇 |
2011年 | 240篇 |
2010年 | 211篇 |
2009年 | 261篇 |
2008年 | 279篇 |
2007年 | 306篇 |
2006年 | 288篇 |
2005年 | 230篇 |
2004年 | 258篇 |
2003年 | 242篇 |
2002年 | 217篇 |
2001年 | 192篇 |
2000年 | 169篇 |
1999年 | 162篇 |
1998年 | 159篇 |
1997年 | 168篇 |
1996年 | 164篇 |
1995年 | 173篇 |
1994年 | 187篇 |
1993年 | 147篇 |
1992年 | 148篇 |
1991年 | 149篇 |
1990年 | 140篇 |
1989年 | 104篇 |
1988年 | 128篇 |
1987年 | 114篇 |
1986年 | 90篇 |
1985年 | 190篇 |
1984年 | 209篇 |
1983年 | 162篇 |
1982年 | 243篇 |
1981年 | 153篇 |
1980年 | 153篇 |
1979年 | 131篇 |
1978年 | 72篇 |
1977年 | 49篇 |
1976年 | 61篇 |
1974年 | 26篇 |
1973年 | 27篇 |
排序方式: 共有8756条查询结果,搜索用时 218 毫秒
181.
The effect of -alany-L-histidinato zinc (AHZ) on bone cell function was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37°C in a CO2 incubator in plastic dishes containing -modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus AHZ (10–7–10–5 M) or other reagents, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10–7–10–5 M) produced a remarkable increase of alkaline phosphatase activity and protein concentration in osteoblastic cells. Thus increases were seen with the prolonged cultivation (12–21 days). With the culture of 1, 3 and 12 days, the effect of AHZ (10–6 M) to increase alkaline phosphatase activity and protein concentration was more intensive than the effect of zinc sulfate, (10–6 M). The AHZ effects were completely abolished by the presence of cycloheximide (10–6 M), indicating that AHZ stimulates protein synthesis in the cells. The present study suggests that AHZ has a stimulatory effect on cell differentiation, and that this effect is partly involved on protein synthesis in osteoblastic cells. 相似文献
182.
Since the positive charge on the lysine residues plays an important role in the receptor recognition ability of oLH, the hormonotoxin has been synthesised with the use of 2-iminothiolane HC1 (2IT) and N-Succinimidyl-3-(2-pyridyldithio)-propionate (SPDP). The oLH activated with 2IT (oLH-10) was then mixed with SPDP activated gelonin (gelonin-30) in order to obtain a oLH-S-S-gelonin hormonotoxin. The conjugation mixture containing hormonotoxin was purified by gel-filtration chromatography according to the molecular weight and a complete physico-chemical, immunochemical and biochemical analysis were performed. The linkage occured through the -NH2 groups of -subunit of oLH as judged from RP-HPLC analysis. A 11 (oLH:gelonin) molar ratio was obtained when determined with the use of several techniques. The hormonotoxins retained substantial receptor binding, steroidogenic activity and immunoreactivity. The competitive displacement analysis indicate that the binding occurs via the hormone part leaving the gelonin free which was probed with the gelonin antibodies. The presently described (C150A-02, C160A-02 and C170A-02) hormonotoxins exhibited higher receptor binding and toxicity to the target cells than the hormonotoxins prepared with the use of SPDP only. Therefore it is concluded that higher receptor binding and cytotoxicity may be due to the retention of positive charge on the lysine residues of oLH which was preserved during the conjugation process.Abbreviations BSA
Bovine Serum Albumin
- CMC
Carboxy methyl Cellulose
- DTT
Dithiothreitol
- DMEM
Dulbeco's Modified Eagle's Medium
- DTNB
Ellman's reagent [5,5-dithio-bis-(2-nitrobenzoic acid)]
- EDTA
Ethylenediaminetetraacetic acid
- FPLC
Fast Protein Liquid Chromatography
- FCA
Freund's Complete Adjuvant
- FCS
Fetal Calf Serum
- Gelonin-30
Gelonin modified by SPDP
- GnRH
Gonadotropin-Releasing Hormone
- Gelonin-SPDP
SPDP modified derivative of gelonin
- HEPES
(N-[2-hydroxyethyl] piperazine-N-[-2-ethanesulphonic acid])
- IFA
Incomplete Freund's Adjuvant
- 2IT
2-Iminothiolane
- IODOGEN
1,3,4,6-tetrachloro 3,6-diphenylglycouril
- oLH
Ovine Luteinizing Hormone
- oLH-SPDP
SPDP modified derivative of oLH
- oLH-10
oLH modified by 2IT
- oLH2IT
Molar ratio of oLH and 2IT
- PDP
2-Pyridyl-dithiopropionate
- PAP
Pokeweed Antiviral Protein
- RIP
Ribosome Inactivating Protein
- RP-HPLC
Reverse-Phase High Performance Liquid Chromatography
- RPMI
Roswell Park Memorial Institute
- RIA
Radioimmunoassay
- RRA
Radioreceptor Assay
- SPDP
N-Succinimidyl-3(2-pyridyldithio)propionate
- SDS-PAGE
Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis
- TCA
Trichloroacetic acid
- TFA
Trifluroacetic acid 相似文献
183.
José L. Sanz Gertrud Huber Harald Huber Ricardo Amils 《Journal of molecular evolution》1994,39(5):528-532
The sensitivity of the cell-free protein synthesis systems from Acidanus brierleyi, Acidianus infernus, and Metallosphaera sedula, members of the archaeal order Sulfolobales, to 40 antibiotics with different specificities has been studied. The sensitivity patterns were compared to those of Sulfolobus solfataricus and other archaeal, bacterial, and eukaryotic systems. The comparative analysis shows that ribosomes from the sulfolobales are the most refractory to inhibitors of protein synthesis described so far. The sensitivity results have been used to ascertain in phylogenetic relationships among the members of the order Sulfolobales. The evolutionary significance of these results are analyzed in the context of the phylogenetic position of this group of extreme thermophilic microorganisms.
Correspondence to: R. Amils 相似文献
184.
Nagayasu T Miyanaga M Tanaka T Sakiyama T Nakanishi K 《Biotechnology and bioengineering》1994,43(11):1108-1117
N-(Benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester (Z-AspPheOMe), a precursor of the aspartame, and N-(benzyloxycarbonyl)-L-phenylalanyl-Lphenylalanine methyl ester (Z-PhePheOMe) were synthesized from the respective amino acid derivatives with an immobilized thermolysin (EC 3.4.24.4) in ethyl acetate. Various factors affecting the synthesis of these dipeptide precursors were clarified. The initial synthetic rate was the highest at the water content of 3.5% for both reactions. The substrate concentration dependencies of the initial synthetic rate of Z-AspkPheOMe and Z-PhePheOMe with the immobilized enzyme in ethyl acetate were different from those in an aqueous buffer solution saturated with ethyl acetate but similar to those in the aqueous/organic biphasic system using the free enzyme. Particularly, the initial synthetic rate of Z-AspPhOMe increased in order higher than first order with respect to the concentration of L-phenylalanine methyl ester (PheOMe), whereas it decreased sharply with the concentration of N-(benzyloxycarbonyl)-L-aspartic acid (Z-Asp). Such kinetic behavior could be explained by regarding the inside of the immobilized enzyme as being a biphasic mode composed from the organic phase and aqueous phase where the enzymatic reaction takes place. The reaction in the aqueous/organic biphasic system using the free enzyme could be simulated by taking into consideration the partition of the substrate and the initial rate of synthesis in the aqueous buffer saturated with ethyl acetate. Based on this analysis, the rate of reaction with the immobilized enzyme in ethyl acetate could also be predicted. Z-AsPheOMe and Z-PhePheOMe were synthesized by the fed-batch method where the acid component of the substrate was intermittently added during the course of reaction and by the batch method. In the synthesis of Z-AspPheOMe, the synthetic rate and maximum yield of reaction as well as the stability of the immobilized enzyme were higher in the fed-batch reaction than those in the batch reaction. In the synthesis of Z-PhePheOMe, the results obtained by both methods were similar. (c) 1994 John Wiley & Sons, Inc. 相似文献
185.
186.
187.
188.
Summary The types and amount of plasma membrane proteins synthesized during cell elongation in response to auxin (2,4-dichlorophenoxyacetic acid) treatment were investigated. Auxin-treated and control soybean (Glycine max L.) hypocotyl segments were incubated with [35S]methionine for various times, ranging from 0.5 to 18 h, prior to isolation of plasma membrane by aqueous two-phase partitioning. Protein accumulated in the plasma membrane after auxin treatment. Despite this accumulation, the protein incorporation rate, estimated by the amount of label in the plasma membrane following a 0.5 h [35S]methionine pulse, was unaffected by auxin treatment at both 0.5 and 18 h of treatment. Protein apparently accumulated by a mechanism distinct from enhanced incorporation. The plasma membrane proteins synthesized by elongating segments differed from controls at 18 h, as evidenced by the pattern of fluorographs following a 0.5 h radiolabelling. However, auxin treatment did not alter the 2-D gel pattern of the polypeptides detectable by silver stain.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- IEF
isoelectric focusing
- PM
plasma membrane
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis 相似文献
189.
Yongqing Liu Jan H. W. Bergervoet C. H. Ric De Vos Henk W. M. Hilhorst H. Lieke Kraak Cees M. Karssen Raoul J. Bino 《Planta》1994,194(3):368-373
The role of cis-abscisic acid (ABA) and gibberellins (GAs) in the induction of cell-cycle activities has been studied during imbibition and subsequent germination of tomato seeds. Using flow cytometry, nuclear replication activity was investigated in embryo root tips isolated from seeds of the ABA-deficient mutant sit w , the GA-deficient mutant gib-1, and the wild-type (MM) tomato (Lycopersicon esculentum Mill. cv. Moneymaker) upon imbibition in water, 10 μM GA4+7, 5 μM ABA or 5 μM ABA+10 μM GA4+7. The nuclei of fully matured dry MM, sit w and gib-1 seeds predominantly showed 2C DNA signals, indicating that the cell-cycle activity of most root-tip cells had been arrested at the G1 phase of nuclear division. However, ABA-deficient sit w seeds contained a significantly higher amount of G2 cells (4C DNA) compared with the other genotypes, suggesting that, during maturation, cell-cycle activity in sit w seeds is less efficiently arrested in G1. Upon imbibition in water, an induction of the 4C signal, indicating nuclear replication, was observed in the root tip cells of both MM and sit w embroys. The augmentation in the 4C signal occurred before visible germination. Gib-1 seeds did not show cell-cycle activity and did not germinate in water. Upon imbibition in GA4+7, both cell-cycle activity and subsequent germination were enhanced in MM and sit w seeds, and were induced in gib-1. In ABA, the germination of MM and sit w seeds was inhibited while nuclear replication of these seeds was not affected. It is concluded that GA influences germination by acting upon processes that precede cell-cycle activation, while ABA affects growth by acting upon processes that follow cell-cycle activation. 相似文献
190.
Selenite has been shown to undergo intracellular metabolism that results in its conversion to other low molecular weight Secontaining
species and also to its incorporation into a selenocysteine residue in selenoprotein. In order to investigate whether the
incorporation into protein is required for the cytotoxic effects of selenite, we have examined whether inhibition of protein
synthesis prevents the inhibitory effect of selenite on the ability of cells to form colonies or to synthesize RNA. We have
found that treatment of HeLa cells with cycloheximide inhibited protein synthesis by >90% but had no effect on the inhibitory
effect of selenite on cell colony formation or RNA synthesis. Since protein synthesis is not necessary for these cytotoxic
effects of selenite they are unlikely to result from an increase in the synthesis of selenoproteins. 相似文献