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91.
J D Morrey S M Bourn T D Bunch M K Jackson R W Sidwell L R Barrows R A Daynes C A Rosen 《Journal of virology》1991,65(9):5045-5051
UV irradiation has been shown to activate the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) in cell culture; however, only limited studies have been described in vivo. UV light has been categorized as UV-A (400 to 315 nm), -B (315 to 280 nm), or -C (less than 280 nm); the longer wavelengths are less harmful but more penetrative. Highly penetrative UV-A radiation constitutes the vast majority of UV sunlight reaching the earth's surface but is normally harmless. UV-B irradiation is more harmful but less prevalent than UV-A. In this report, the HIV-1 LTR-luciferase gene in the skin of transgenic mice was markedly activated when exposed to UV-B irradiation. The LTR in the skin of transgenic mice pretreated topically with a photosensitizing agent (psoralen) was also activated to similar levels when exposed to UV-A light. A 2-h exposure to sunlight activated the LTR in skin treated with psoralen, whereas the LTR in skin not treated with psoralen was activated after 7 h of sunlight exposure. The HIV-1 LTR-beta-galactosidase reporter gene was preferentially activated by UV-B irradiation in a small population of epidermal cells. The transgenic mouse models carrying HIV-1 LTR-luciferase and LTR-beta-galactosidase reporter genes have been used to demonstrate the in vivo UV-induced activation of the LTR and might be used to evaluate other environmental factors or pharmacologic substances that might potentially activate the HIV-1 LTR in vivo. 相似文献
92.
T.D. Morgan A.E. Beezer J.C. Mitchell & A.W. Bunch 《Journal of applied microbiology》2000,90(1):53-58
T. D. MORGAN, A. E. BEEZER, J. C. MITCHELL AND A. W. BUNCH. 2001 .
Aims: This study aimed to evaluate the efficacy of 'natural' putative antimicrobial agents against Streptococcus mutans and to compare these with synthetic agents using the flow microcalorimeter. Streptococcus mutans is one of the oral pathogens responsible for dental caries.
Methods and Results: Traditional microbiological techniques are invasive and destructive unlike flow microcalorimetry. This rapid technique was used to continuously monitor the power output (bioactivity) of Strep. mutans with reproducibility, precision and accuracy. The antibacterial agents found in oral hygiene products and all the natural agents tested showed anti- Strep. mutans ability.
Conclusions: In this study microcalorimetry identified agents that had a biological effect and quantified the rate of kill achieved enabling four broad categories of antimicrobial agent to be defined.
Significance and Impact of the Study: Microcalorimetric data are a better indication of antimicrobial efficacy than merely determining concentrations at which an antimicrobial agent is bacteriostatic or bactericidal. 相似文献
Aims: This study aimed to evaluate the efficacy of 'natural' putative antimicrobial agents against Streptococcus mutans and to compare these with synthetic agents using the flow microcalorimeter. Streptococcus mutans is one of the oral pathogens responsible for dental caries.
Methods and Results: Traditional microbiological techniques are invasive and destructive unlike flow microcalorimetry. This rapid technique was used to continuously monitor the power output (bioactivity) of Strep. mutans with reproducibility, precision and accuracy. The antibacterial agents found in oral hygiene products and all the natural agents tested showed anti- Strep. mutans ability.
Conclusions: In this study microcalorimetry identified agents that had a biological effect and quantified the rate of kill achieved enabling four broad categories of antimicrobial agent to be defined.
Significance and Impact of the Study: Microcalorimetric data are a better indication of antimicrobial efficacy than merely determining concentrations at which an antimicrobial agent is bacteriostatic or bactericidal. 相似文献
93.
Zhiqiang Fan Wei Li Sang R. Lee Qinggang Meng Bi Shi Thomas D. Bunch Kenneth L. White Il-Keun Kong Zhongde Wang 《PloS one》2014,9(10)
The golden Syrian hamster is the model of choice or the only rodent model for studying many human diseases. However, the lack of gene targeting tools in hamsters severely limits their use in biomedical research. Here, we report the first successful application of the CRISPR/Cas9 system to efficiently conduct gene targeting in hamsters. We designed five synthetic single-guide RNAs (sgRNAs)—three for targeting the coding sequences for different functional domains of the hamster STAT2 protein, one for KCNQ1, and one for PPP1R12C—and demonstrated that the CRISPR/Cas9 system is highly efficient in introducing site-specific mutations in hamster somatic cells. We then developed unique pronuclear (PN) and cytoplasmic injection protocols in hamsters and produced STAT2 knockout (KO) hamsters by injecting the sgRNA/Cas9, either in the form of plasmid or mRNA, targeting exon 4 of hamster STAT2. Among the produced hamsters, 14.3% and 88.9% harbored germline-transmitted STAT2 mutations from plasmid and mRNA injection, respectively. Notably, 10.4% of the animals produced from mRNA injection were biallelically targeted. This is the first success in conducting site-specific gene targeting in hamsters and can serve as the foundation for developing other genetically engineered hamster models for human disease. 相似文献
94.
Tri H.V. Huynh Bjarke Abrahamsen Karsten K. Madsen Alba Gonzalez-Franquesa Anders A. Jensen Lennart Bunch 《Bioorganic & medicinal chemistry》2012,20(23):6831-6839
The excitatory amino acid transporters (EAATs) play a pivotal role in regulating the synaptic concentration of glutamate in the mammalian central nervous system. To date, five different subtypes have been identified, named EAAT15 in humans (and GLAST, GLT-1, EAAC1, EAAT4, and EAAT5, respectively, in rodents). Recently, we have published and presented a structure–activity relationship (SAR) study of a novel class of selective inhibitors of EAAT1 (and GLAST), with the analogs UCPH-101 (IC50 = 0.66 μM) and UCPH-102 (IC50 = 0.43 μM) being the most potent inhibitors in the series. In this paper, we present the design, synthesis and pharmacological evaluation of six coumarin-based fluorescent analogs of UCPH-101/102 as subtype-selective inhibitors at EAAT1. Analogs 1114 failed to inhibit EAAT1 function (IC50 values >300 μM), whereas analogs 15 and UCPH-102F inhibited EAAT1 with IC50 values in the medium micromolar range (17 μM and 14 μM, respectively). Under physiological pH no fluorescence was observed for analog 15, while a bright blue fluorescence emission was observed for analog UCPH-102F. Regrettably, under confocal laser scanning microscopy selective visualization of expression of EAAT1 over EAAT3 was not possible due to nonspecific binding of UCPH-102F. 相似文献
95.
Sera from 287 sheep were screened for cytotoxic antibodies against sheep lymphocytes. Forty four antisera were selected which provisionally define 13 lymphocyte antigens. The frequency of these antigens was studied in 305 sheep from 8 flocks of different breeds. Family studies confirm that inheritance of sheep lymphocyte antigens is controlled by the autosomal codominant genes of at least 2 linked loci. 相似文献
96.
Philippe Bridonneau Steve Bunch Robert Tengler Ken Hill Jeff Carter Wolfgang Pieken David Tinnermeier Russ Lehrman Daniel W. Drolet 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1999,726(1-2)
To evaluate RNA-aptamers as potential drug candidates, efficient and scaleable purification protocols are needed. Because aptamers are highly structured and rigid molecules, denaturation during the purification process is a critical aspect to obtain a pure and active product. A two-step chromatographic procedure was developed to purify a synthetic anti-VEGF aptamer at the preparative scale. A reversed-phase chromatographic step was optimized with a highly hydrophobic ion pairing reagent, followed by ion-exchange chromatography in which heat and a chaotropic salt were used. Because of the presence of 2′-modified ribose, denaturation conditions had to be optimized in both chromatographic steps to achieve a fully active molecule. 相似文献
97.
Helsten TL Bunch TA Kato H Yamanouchi J Choi SH Jannuzi AL Féral CC Ginsberg MH Brower DL Shattil SJ 《Molecular biology of the cell》2008,19(8):3589-3598
Integrin-mediated cell adhesion is essential for development of multicellular organisms. In worms, flies, and vertebrates, talin forms a physical link between integrin cytoplasmic domains and the actin cytoskeleton. Loss of either integrins or talin leads to similar phenotypes. In vertebrates, talin is also a key regulator of integrin affinity. We used a ligand-mimetic Fab fragment, TWOW-1, to assess talin's role in regulating Drosophila alphaPS2betaPS affinity. Depletion of cellular metabolic energy reduced TWOW-1 binding, suggesting alphaPS2betaPS affinity is an active process as it is for vertebrate integrins. In contrast to vertebrate integrins, neither talin knockdown by RNA interference nor talin head overexpression had a significant effect on TWOW-1 binding. Furthermore, replacement of the transmembrane or talin-binding cytoplasmic domains of alphaPS2betaPS with those of human alphaIIbbeta3 failed to enable talin regulation of TWOW-1 binding. However, substitution of the extracellular and transmembrane domains of alphaPS2betaPS with those of alphaIIbbeta3 resulted in a constitutively active integrin whose affinity was reduced by talin knockdown. Furthermore, wild-type alphaIIbbeta3 was activated by overexpression of Drosophila talin head domain. Thus, despite evolutionary conservation of talin's integrin/cytoskeleton linkage function, talin is not sufficient to regulate Drosophila alphaPS2betaPS affinity because of structural features inherent in the alphaPS2betaPS extracellular and/or transmembrane domains. 相似文献
98.
The distribution and prevalence of mandibular osteomyelitis, lumpy jaw, and other dental anomalies in wild sheep were investigated and their biological and evolutionary implications were assessed. Our survey was based on 3,363 mandibles of wild sheep and 1,028 from domesticated varieties. Lumpy jaw is widespread in wild sheep of North America, but it is rare or absent in wild sheep from Eurasia. Among the subspecies of Ovis spp. in North American, the thinhorn sheep (Ovis dalli) were the most seriously impacted, with a prevalence in Dall's sheep (O. dalli dalli) of 23.3% and 29.3% in Stone's sheep (O. dalli stonei). Among the bighorns (O. canadensis), the Rocky Mountain subspecies (O. canadensis canadensis) had a higher rate (12.1%) than other subspecies. Lumpy jaw was not documented in the desert sheep of Baja California (O. canadensis cremnobates, O. canadensis weemsii). Based on data from affected thinhorn sheep, it appears there is an inverse relationship between age of a subspecies in a long term evolutionary context and susceptibility to lumpy jaw. In Eurasian wild sheep lumpy jaw is rare or absent with prevalences ranging from 0 to 7.1% among suspecies, and in domesticated breeds the prevalence averaged 5.0%. The impact of lumpy jaw on different age classes or longevity is equivocal, although females are more susceptible than males. Lumpy jaw appears to effect horn development in males. 相似文献
99.
Wang S Foote WC Sutton DL Maciulis A Miller JM Evans RC Holyoak GR Call JW Bunch TD Taylor WD Marshall MR 《Theriogenology》2001,56(2):315-327
This study investigated whether the transmission of naturally occurring scrapie in sheep can be prevented using embryo transfer. Embryos were collected from 38 donor ewes in a Suffolk sheep flock with a high incidence of naturally occurring scrapie, treated with a sanitary procedure (embryo washing) recommended by the International Embryo Transfer Society and then transferred to 58 scrapie-free recipient ewes. Ninety-four offspring were produced. None of the offspring or the recipient ewes developed scrapie. Furthermore, offspring derived from embryos collected from donor ewes bred to the immunohistochemically positive ram did not develop scrapie. We conclude that scrapie was not transmitted to offspring via the embryo nor was the infective agent transmitted to recipient ewes during embryo transfer procedures. 相似文献
100.
Li GP Bunch TD White KL Aston KI Meerdo LN Pate BJ Sessions BR 《Molecular reproduction and development》2004,68(2):189-197
Treatment of in vitro matured bovine oocytes with colcemid results in a membrane protrusion that contains maternal chromosomes, which can be easily removed by aspiration. Four experiments were designed to evaluate the overall and temporal effects of conditioned medium (CM) by bovine cumulus cells on development of nuclear transfer (NT) bovine embryos and to examine the chromosomal composition and allocation of inner cell mass (ICM) and trophectoderm (TE) of the subsequent blastocysts. The nuclear transfer embryos were cultured in various CR1aa media conditioned by preculture with bovine cumulus cells. Development to the blastocyst stage in BSA-containing CM (BCM) and serum-containing CM (SCM) were similar to co-culture group (24-30%). The 24 hr-conditioned BCM yielded higher blastocyst development than 48 and 72 hr-conditioned BCM. Temporary exposure of embryos to BCM and SCM followed by CR1aa was also studied. Morula and blastocyst development were not different among the groups cultured in BCM for 72, 96, and 168 hr, but were significantly higher (P < 0.01) than groups exposed to BCM for 24 and 48 hr, respectively. Blastocyst development in SCM for 24 hr (29%), 96 hr (25%), and 168 hr (27%) were much higher (P < 0.05) than those in SCM for 48 hr (12%) and 72 hr (10%). The analyses of chromosomal composition of the resulting blastocysts indicate approximately 80% of the blastocysts cultured in CR1aa with co-culture or groups initially exposed to BCM for 24 hr followed by culture in CR1aa were diploid. However, the incidence of diploidy were only 36-60% in SCM-cultured groups and groups cultured in BCM beyond 48 hr. Conditioned media did not affect the allocation of ICM and TE in the blastocyst. No difference was found in the ratio of inner cell mass to total cells in co-culture, BCM or SCM groups (0.424, 0.441, and 0.473, respectively). In conclusion, bovine cumulus cell-CM and CR1aa with co-culture supported comparable development and blastocyst ICM:total cell ratio of bovine NT embryos. However, CM affected the blastocyst chromosomal composition and induced higher mixploidy. 相似文献