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71.
Isolation of chromosome-21-specific DNA probes and their use in the analysis of nondisjunction in Down syndrome 总被引:8,自引:2,他引:6
Summary Thirteen single-copy, chromosome-21-specific DNA probes were isolated from a recombinant library made from flow-sorted chromosome 21 DNA and regionally mapped using a panel of somatic cell hybrids. Five probes mapped in the 21q21-q22.1 region, six to the 21q22.1-qter region, and one to each of the regions 21q22.1-q22.2 and 21q22.3. Two of these probes, one of which maps in the critical region for Down syndrome, have recently been shown to be expressed at high levels in Down syndrome brain tissue (Stefani et al. 1988). Following preliminary screening for restriction fragment lenght polymorphisms (RFLPs), five polymorphisms were discovered with four of the chromosome 21 DNA probes. A frequent MspI polymorphism detected by one of the probes was used in conjunction with four previously described polymorphic chromosome 21 probes to analyse the origin of nondisjunction in 33 families with a child or fetus with trisomy 21. The parental origin of the additional chromosome 21 was determined in 12 cases: in 9 (75%) of these it was derived from the mother and in the other 3 cases (25%) it was of paternal origin. Cytogenetic analysis of Q-banding heteromorphisms was informative in three of five families tested, and in each case the RFLP results were confirmed. The meiotic stage of nondisjunction was defined with confidence in five families, the results being obtained with pericentromeric RFLP or cytogenetic markers. Recombination between two nondisjoined chromosomes was demonstrated in one family and is consistent with the view that a lack of recombination between chromosome 21 homologues or failure of their conjunction is not the invariable cause of trisomy 21. 相似文献
72.
Photoaffinity labeling with [32P] 8-azidoadenosine 5-triphosphate (8-N3ATP) was used to identify putative binding sites on tobacco (Nicotiana tabacum L. and N. rustica L.) leaf ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCase, EC 4.1.1.39). Incorporation of 32P was observed in polypeptides corresponding to both RuBPCase subunits when desalted leaf and chloroplast extracts, and purified RuBPCase were irradiated with ultraviolet light in the presence of [32P] 8-N3ATP. 32P-labeling was dependent upon ultraviolet irradiation and occurred with [32P] 8-N3ATP labeled in the -position, indicating covalent incorporation of the photoprobe. Both [32P] 8-N3ATP and [32P] 8-N3GTP were incorporated to a similar extent into the 53-kilodalton (kDa) large subunit (LSu), but incorporation of [32P] 8-N3GTP into the 14-kDa small subunit (SSu) of RuBPCase was <5% of that measured with [32P] 8-N3ATP. Distinct binding sites for 8-N3ATP on the two subunits were indicated by different apparent K
D
values, 3 and 18 M for the SSu and LSu, respectively, and differences in the response of photoaffinity labeling to Mg2+, anions and enzyme activation. Active-site-directed compounds, including the non-gaseous substrate ribulose 1,5-bisphosphate, the reaction intermediate analog 2-carboxyarabinitol-1,5-bisphosphate and several phosphorylated effectors afforded protection to the LSu site against photoincorporation but provided almost no protection to the SSu. These results indicate that 8-N3ATP binds to the active-site region of the LSu and a distinct site on the SSu of RuBPCase. Experiments conducted with intact pea (Pisum sativum L.) and tobacco chloroplasts showed that the SSu was not photolabeled with [32P] 8-N3ATP in organello or in undesalted chloroplast lysates but was photolabeled when lysates were ultrafiltered or desalted. These results indicate that 8-N3ATP binds to a site on the SSu that has physiological significance.Abbreviations kDa
kilodalton
- LSu
large subunit
- 8-N3ATP
8-azidoadenosine 5-triphosphate
- RuBP
ribulose-1,5-bisphosphate
- RuBPCase
ribulose-1,5-bisphosphate carboxylase/oxygenase
- SSu
small subunit
Kentucky Agricultural Experiment Station Journal Article No. 89-3-150The authors acknowledge the technical assistance of J.C. Anderson. This work was supported in part by National Institute of Health grant GM 35766 to B.E.H. 相似文献
73.
The radial forearm flap: reconstructive applications and donor-site defects in 35 consecutive patients 总被引:5,自引:0,他引:5
Thirty-five consecutive patients treated with the radial forearm flap were reviewed. This flap was used in head and neck reconstruction in 25 patients, soft-tissue cover of an extremity in 9 patients, and as a new technique for penile reconstruction in 1 patient. Osteocutaneous flaps were used for mandibular reconstruction in 13 patients. In 6 patients innervated flaps were used to provide sensation on the dorsum of the hand or on the weight-bearing surface of the foot. There was only one total flap failure and no partial failures. Recipient-site complications were few, with prompt healing and very acceptable appearance. Donor-site complications included partial loss of the skin graft with tendon exposure in 10 patients (33 percent), an unsatisfactory appearance in 5 patients (17 percent), and one case of radial fracture (8 percent). On functional testing, there was no significant loss of strength or joint mobility in the donor extremity in 19/20 patients. The authors recommend measures to reduce donor-site morbidity and conclude that, with an acceptable donor site, this flap is valuable in a variety of reconstructive applications. 相似文献
74.
Surgical augmentation of skin blood flow and viability in a pig musculocutaneous flap model 总被引:2,自引:0,他引:2
J B Boyd B Markland D Dorion C Y Pang S Morris 《Plastic and reconstructive surgery》1990,86(4):731-738
A porcine rectus abdominis musculocutaneous (TRAM) flap model was designed and validated in nine pigs. This TRAM flap was based on the deep inferior epigastric (DIE) vessels with an 8 x 18 cm transverse skin paddle at the superior end of the rectus abdominis muscle. The model was subsequently used to test our hypothesis of surgical augmentation of flap viability by vascular territory expansion. Specifically, we observed that ligation of the superior epigastric (SE) vessels at 4, 7, 14, and 28 days (N = 6 to 8) prior to raising the TRAM flaps significantly increased (p less than 0.05) the length and area of the viable skin in the transverse skin paddles of the treatment flaps compared with the contralateral shammanipulated control flaps. This significant increase in skin viability was seen to be accompanied by a significant increase (p less than 0.05) in skin and muscle capillary blood flow in the treatment TRAM flaps compared with the controls (N = 9). The mechanism of vascular territory expansion is unclear. We postulate that hypoxia resulting from the ligation of the superior epigastric vessels prior to the flap surgery may play a role in the triggering of the deep inferior epigastric artery to take over some of the territory previously perfused by the superior epigastric artery. This would then increase the skin and muscle capillary blood flow in the transverse paddle when the TRAM flap was raised on the deep inferior epigastric vascular pedicle. 相似文献
75.
A new chemical method for carboxy-terminal (C-terminal) protein sequencing has been developed. This approach has been successfully used to sequence 5 residues of standard proteins and 5 to 10 residues of synthetic peptides at low nanomole levels. The sequencing procedure consists of converting the C-terminal amino acid into a thiohydantoin (TH) derivative, followed by transformation of the TH into a good leaving group by alkylation. Next, the alkylated TH is cleaved mildly and efficiently with (N = C V S)- anion, which simultaneously forms a TH on the newly truncated protein or peptide. Thus, after the initial TH derivatization, there is no return to a free carboxyl group at the C-terminus. An additional benefit of this method is that the alkylating moiety can be chosen with a variety of properties allowing for variation in the detection method. This chemistry has been adapted to automated protein sequencers with a cycle time of about 1 h. 相似文献
76.
Establishment of polychlorinated biphenyl-degrading enrichment culture with predominantly meta dechlorination. 总被引:4,自引:4,他引:0 下载免费PDF全文
Enrichment of polychlorinated biphenyl (PCB)-dechlorinating microorganisms from PCB-contaminated sediments from the Upper Hudson River, N.Y., was attempted. The enrichment strategy was to use pyruvate as the electron donor and dechlorination of Aroclor 1242 as the electron acceptor. The enrichment medium also contained non-PCB-contaminated Hudson River sediments, which were required for the PCB-dechlorinating activity. An enrichment culture (that had stable PCBT-dechlorinating activity over nine serial transfers during 1 year) was established under these conditions; however, the rate of dechlorination did not increase after the second serial transfer. Dechlorination occurred primarily from the meta positions of the biphenyl molecule. Hydrogen could be substituted for pyruvate as the electron donor with equal activity, but when acetate was used as the electron donor a delay in dechlorination was observed. Sulfate and bromethane sulfonate inhibited dechlorination activity. The pyruvate-Aroclor 1242 enrichment also dechlorinated Aroclors 1248, 1254, and 1260; the extent of chlorine removed was the greatest for Aroclor 1254. For comparison, nonautoclaved non-PCB-contaminated Hudson River sediments used in the assay also dechlorinated Aroclors, but only after 12 to 16 weeks of incubation. This suggests that PCB-dechlorinating organisms were also present in these sediments but in numbers lower than those in the enrichment culture. 相似文献
77.
The initial rate of transport of the bile acid glycocholic acid (GCA) has been measured in influx and efflux across placental basal membrane vesicles, and the mechanism of inhibition of its transport by the analogue taurochenodeoxycholic acid (TCDCA) analysed kinetically. This analogue, although trans-stimulating GCA efflux, inhibits influx in a way which does not depend upon substrate concentration; moreover, its potency as an inhibitor is markedly influenced by whether it is placed on one or on both sides of the vesicles membrane. These findings can be accounted for by postulating that both GCA and TCDCA are translocated through the carrier, but that the rate of loaded carrier reorientation is higher than that of the free carrier only when loaded with TCDCA and not with GCA. 相似文献
78.
中国蝾螈科动物的反捕行为(正文) 总被引:1,自引:0,他引:1
中国的蝾螈科(Family Salamandridae)动物演化出了卓越的,多种多样的特征,而这些特征往往伴随着强烈的皮肤毒腺存在来保护自己免受捕食者的侵害。这些反捕食行为机制分布列入表1,并以照片解释其反捕行为,来用于已创建的定义和术语是非常重要的。这些定义和术语与不同反捕行为是密切相关的。该科中大多数蝾螈都有鲜艳的腹部色型,有皮肤分泌物毒素存在以警告捕食者。有的蝾螈,象兰尾蝾螈Cynops cyanura和中国瘰螈Paramesotriton chinensis甚至向上翻转显示腹部颜色。 相似文献
79.
In order to advance knowledge of the neural control of feeding,we investigated the cortical representation of the taste oftannic acid, which produces the taste of astringency. It isa dietary component of biological importance particularly toarboreal primates. Recordings were made from 74 taste responsiveneurons in the orbitofrontal cortex. Single neurons were foundthat were tuned to respond to 0.001 M tannic acid, and representeda subpopulation of neurons that was distinct from neurons responsiveto the tastes of glucose (sweet), NaCl (salty), HCI (sour),quinine (bitter) and monosodium glutamate (umami). In addition,across the population of 74 neurons, tannic acid was as wellrepresented as the tastes of NaCI, HCI quinine or monosodiumglutamate. Multidimensional scaling analysis of the neuronalresponses to the tastants indicates that tannic acid lies outsidethe boundaries of the four conventional taste qualities (sweet,sour, bitter and salty). Taken together these data indicatethat the astringent taste of tannic acid should be consideredas a distinct taste quality, which receives a separate representationfrom sweet, salt, bitter and sour in the primate cortical tasteareas. Chem. Senses 21: 135145, 1996. 相似文献
80.
Chilling ofArabidopsis thaliana (L.) Heynh. callus tissue to 4 °C led to conditions of oxidative stress, as indicated by increased levels of the products of peroxidative damage to cell membranes. Cellular H2O2 was also observed to increase initially upon chilling but by day 8 cellular levels had declined to below control levels. Although levels of catalase activity remained similar to those in unchilled tissue, activity of ascorbate peroxidase increased between days 4 and 8 of chilling to 4 °C. In callus held at 23 °C, levels of reduced glutathione remained static whereas they rose in callus held at 4 °C. Levels of oxidised glutathione were initially low but increased significantly by day 4 in the chilled callus. At 23 °C, however, levels of oxidised glutathione remained low. Between days 1 and 3 at 4 °C, levels of glutathione reductase activity increased but by day 8 glutathione reductase activity was similar to that in cells held at 23 °C. Exposure of callus to abscisic acid at 23 °C also led to increased activities of ascorbate peroxidase and glutathione reductase.Abbreviations ABA
abscisic acid
- GSH
reduced glutathione
- GSSG
oxidised glutathione
- TTC
2,35-triphenyltetrazolium chloride
This work is supported by a grant from the Biotechnology and Biological Sciences Research Council. 相似文献