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41.
Esterases and lipases can process amphiphilic esters used as drugs and prodrugs and impact their pharmacokinetics and biodistribution. These hydrolases can also process ester components of drug delivery systems (DDSs), thus triggering DDSs destabilization with premature cargo release. In this study we tested and optimized assays that allowed us to quantify and compare individual esterase contributions to the degradation of substrates of increased lipophilicity and to establish limitations in terms of substrates that can be processed by a specific esterase/lipase. We have studied the impact of carbonic anhydrase; phospholipases A1, A2, C and D; lipoprotein lipase; and standard lipase on the hydrolysis of 4-nitrophenyl acetate, 4-nitrophenyl palmitate, DGGR and POPC liposomes, drawing structure–property relationships. We found that the enzymatic activity of these proteins was highly dependent on the lipophilicity of the substrate used to assess them, as expected. The activity observed for classical esterases was diminished when lipophilicity of the substrate increased, while activity observed for lipases generally increased, following the interfacial activation model, and was highly dependent on the type of lipase and its structure. The assays developed allowed us to determine the most sensitive methods for quantifying enzymatic activity against substrates of particular types and lipophilicity. 相似文献
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43.
The mutagenicity response of well-characterized flash vacuum thermolysis (FVT) pyrolysates that contain cyclopenta[cd]pyrene (1) and 1-ethynylpyrene (1b) (pyrolysate I), the dicyclopentapyrene congeners dicyclopenta[cd,mn]- (2), dicyclopenta[cd,fg]- (3), or dicyclopenta[cd,jk]pyrene (4) and their related bis-ethynyl- (2b–4b) and monocyclopenta-ethynylpyrene (2a–4a) analogues (pyrolysates II–IV, respectively), or cyclopenta[cd]- (1) and the three dicyclopentapyrenes (2–4) (pyrolysate V), respectively, was assessed using the standard protocol outlined by Ames et al. (Salmonella typhimurium strain TA98 ± S9-mix 4% (v/v)). It is shown that the mutagenic activity of the pyrolysates deviates from the weighed sum of the activity of the individual pyrolysate constituents. Hence, FVT-pyrolysates are proposed as model mixtures, that is, as partial combustion exhaust mimics, to establish and evaluate interactions (additive, synergistic, or antagonistic effects) that may occur among the constituents and affect the global mutagenicity response. 相似文献
44.
目的以结核分枝杆菌(Mycobacterium tuberculosis,MTB)RD1区特异性培养滤液蛋白10(culture filtrate pro-tein10,CFP10)与戊糖-5-磷酸-3-差异构象酶68(pentose-5-phosphate-3-epimerase68,PPE68)的融合蛋白CFP10-PPE68作为包被抗原,建立检测结核病患者血清中MTB抗体的间接ELISA法。方法采用亲和层析法纯化重组CFP10-PPE68、CFP10和PPE68蛋白;分别以3种蛋白作为包被抗原,建立检测结核病患者血清中MTB特异性抗体的间接ELISA法;采用方阵滴定法对建立的间接ELISA法的条件进行优化;并以临床诊断为金标准,对间接ELISA法的特异性、敏感性和准确性进行验证。结果纯化的重组CFP10-PPE68融合蛋白纯度约为70%。分别以重组CFP10-PPE68、CFP10和PPE68蛋白作为包被抗原的最佳包被浓度分别为2、4、4μg/ml,血清最佳稀释度均为1∶1 000,酶标二抗最佳稀释度均为1∶5 000。3种蛋白对肺外结核的诊断效果均优于肺结核,且重组CFP10-PPE68融合蛋白检测肺外结核的特异性、敏感性及准确性均最佳;3种蛋白诊断结核病的特异性较高,而敏感性相对较低。结论以重组CFP10-PPE68融合蛋白作为包被抗原建立的间接ELISA法诊断MTB感染具有较高的敏感性和特异性,特别是对难以诊断的肺外结核病的诊断有一定的参考价值。 相似文献
45.
Phungjai Boonyeun Artiwan Shotipruk Chattip Prommuak Manop Suphantharika 《Chemical Engineering Communications》2013,200(12):1594-1602
A two-step autolysis process was proposed to enhance amino acid production from spent brewer's yeast. The technique was developed based on comparative study of the dynamics of production and release of proteins and amino acids during the autolysis of a concentrated suspension (22 wt.%) and a dilute yeast cell suspension (11.25 wt.%). The results suggest that, in the concentrated yeast suspension, proteins are more effectively broken down into amino acids, but the product release rate was lower due to a lower concentration gradient across the cell membrane. Thus, a two-step process, in which a high protein conversion occurred in a concentrated cell suspension during the first 13 h period, followed by a 26 h autolysis process within a dilute cell suspension, provided a higher overall yield of amino acids compared than the single-step process. The two-step process was found to result in a 25% higher amino acid yield with a weight fraction increase from 0.4 to 0.5 g/g dry wt. Other than these findings, the effect of adding NaCl to the suspension during autolysis was also investigated. It was found that, for the autolysis conditions employed in this study, the addition of NaCl did not significantly affect the production of protein but inhibited the production of amino acids. 相似文献
46.
47.
《Journal of Sulfur Chemistry》2013,34(3):240-250
A series of novel 2-aryl1-4-bromothiophenes (2a–f), biarylthiophenes with non-identical aryl groups (3a–e) and biarylthiophenes with identical aryl groups (4a–f) were synthesized in moderate to excellent yields by using different arylboronic acids in a Suzuki–Miyaura cross-coupling reaction. The experimental results showed that the use of K2CO3 as base resulted in moderate yields compared with that of good yields obtained upon using K3PO4. The highest yield obtained using K3PO4 was 82% for 2, 4-bis (4-chlorophenyl) thiophene (4d). The synthesized compounds in the present study were examined for their biofilm inhibition and hemolysis assay. Among all compounds 2, 4-bis (4-methoxyphenyl) thiophene (4b) was found to strongly inhibit the formation of bacterial biofilm against E. coli. The compound 4b exhibited higher inhibition (80.92%) compared with the standard Rifampicin with 97.43% inhibition. The compound 2, 4-bis (4-chlorophenyl) thiophene (4d) displayed strong anti-biofilm activity with its ability to prevent the formation of Pasteurell amultocida biofilm at the percent inhibition of 74.53%. In addition, 2f showed the highest percentage hemolysis 16.0% compared with that of the standard Triton-X-100. 3 相似文献
48.
49.
Selective and pH-Dependent Binding of a Moth Pheromone to a Pheromone-Binding Protein 总被引:1,自引:0,他引:1
Fluorescence and circular dichroism (CD) data suggest that the major pheromone-binding protein (PBP) from the wild silkmoth,
Antheraea polyphemus, ApolPBP1, undergoes a pH-dependent conformational change similar to that previously observed for the PBP from the silkworm
moth, Bombyx mori, BmorPBP. All three constituents of the sex pheromone, E6,Z11-16Ac, E6,Z11-16Ald, and E4,Z9-14Ac, bound to ApolPBP1 with
apparent high affinity at high pH, but reduced binding at low pH when tested individually in a “cold binding assay.” In competitive
assays, however, ApolPBP1 showed considerable preference for the major constituent of the sex pheromone, E6,Z11-16Ac. These
data suggest that specificity of PBPs contributes at least in part to the remarkable selectivity of moth's olfactory system. 相似文献
50.
Adsorption of heavy metals by brewery biomass 总被引:5,自引:0,他引:5
Tae-Young Kim Sun-Kyu Park Sung-Yong Cho Hwan-Beom Kim Yong Kang Sang-Done Kim Seung-Jai Kim 《Korean Journal of Chemical Engineering》2005,22(1):91-98
In this work, biosorption of lead, copper and cadmium by waste brewery yeast has been studied. The adsorption capacity for
lead, copper and cadmium on the biomass increased with the increasing temperature and the maximum uptakes were 0.465 mmolPb/g
(96.4 mg/g), 0.769 mmolCu/g (48.9 mg/g) and 0.127 mmolCd/g (14.3 mg/g) at 308 K. The Langmuir isotherm, favorable type, and
the pseudo second-order kinetic model represent our experimental data very well. The heat of biosorption was evaluated from
the Langmuir isotherm equation, and the biosorption of lead, copper and cadmium was endothermic reaction. 相似文献