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103.
Hideki Kajiura Ryo Kakutani Tsunehisa Akiyama Hiroki Takata Takashi Kuriki 《Biocatalysis and Biotransformation》2013,31(1-2):133-140
Two well-established methods to prepare glycogen are available: (1) extraction from natural resources such as shellfish and animal tissues; (2) synthesis from glucose-1-phosphate using two enzymes, α-glucan phosphorylase (EC 2.4.1.1) and branching enzyme (EC 2.4.1.18). We have developed a novel enzymatic process for glycogen production, in which short-chain amylose is first prepared from starch or dextrin by using isoamylase (EC 3.2.1.68), and then branching enzyme and amylomaltase (EC 2.4.1.25) are added to synthesize glycogen. Our enzymatic process, using isoamylase, branching enzyme and amylomaltase, is currently the most efficient for glycogen production. Furthermore, the molecular weight of glycogen is controllable in a range of 3.0×106 to 3.0×107 by adjusting some parameters of the reaction. 相似文献
104.
Kohji Ichimori Naoto Fukuyama Hiroe Nakazawa Yasuaki Aratani Hideki Koyama Shunya Takizawa 《Free radical research》2013,47(5):481-489
Myeloperoxidase (MPO) catalyzes a nitration reaction to form nitrotyrosine in the presence of high nitrite, the metabolite of NO. Human leukocyte was shown to cause phenolic nitration using released MPO as a catalyst in the presence of nitrite. It opposes our previous finding that inhibition of MPO was essential for phenol nitration in human leukocyte study. To clarify the role of MPO, we utilized MPO-deficient human leukocytes and MPO-knockout mice. Even in the absence of exogenously added nitrite, high nitration product was observed in MPO-deficient leukocytes. In liver subjected to ischemia/reperfusion injury, a significantly higher amount of nitrotyrosine was produced in MPO-knockout mice than in normal mice. These results clearly demonstrate that MPO inhibits the accumulation of nitration products in vivo . Further experiments showed that MPO could degrade nitrotyrosine in the presence of glutathione. Thus, MPO-induced degradation of nitration products may cause the underestimation of the nitration product generated in vivo . We conclude that MPO may act predominantly to scavenge nitrotyrosine under physiological nitrite condition, and protect against injurious effect of nitrotyrosine. 相似文献
105.
Toshio Naka Takashi Doi Takeshi Tsujino Tohru Masuyama Seinosuke Kawashima 《Preparative biochemistry & biotechnology》2013,43(2):87-99
Abstract Viral vector systems are efficient for transfection of foreign genes into many tissues. Especially, retrovirus based vectors integrate the transgene into the genome of the target cells, which can sustain long term expression. However, it has been demonstrated that the transduction efficiency using retrovirus is relatively lower than those of other viruses. Ultrasound was recently reported to increase gene expression using plasmid DNA, with or without, a delivery vehicle. However, there are no reports, which show an ultrasound effect to retrovirus‐mediated gene transfer efficiency. Retrovirus‐mediated gene transfer systems were used for transfection of 293T cells, bovine aortic endothelial cells (BAECs), rat aortic smooth muscle cells (RASMCs), and rat skeletal muscle myoblasts (L6 cells) with β‐galactosidase (β‐Gal) genes. Transduction efficiency and cell viability assay were performed on 293T cells that were exposed to varying durations (5 to 30 seconds) and power levels (1.0 watts/cm2 to 4.0 watts/cm2) of ultrasound after being transduced by a retrovirus. Effects of ultrasound to the retrovirus itself was evaluated by transduction efficiency of 293T cells. After exposure to varying power levels of ultrasound to a retrovirus for 5 seconds, 293T cells were transduced by a retrovirus, and transduction efficiency was evaluated. Below 1.0 watts/cm2 and 5 seconds exposure, ultrasound showed increased transduction efficiency and no cytotoxicity to 293T cells transduced by a retrovirus. Also, ultrasound showed no toxicity to the virus itself at the same condition. Exposure of 5 seconds at the power of 1.0 watts/cm2 of an ultrasound resulted in significant increases in retrovirus‐mediated gene expression in all four cell types tested in this experiment. Transduction efficiencies by ultrasound were enhanced 6.6‐fold, 4.8‐fold, 2.3‐fold, and 3.2‐fold in 293T cells, BAECs, RASMCs, and L6 cells, respectively. Furthermore, β‐Gal activities were also increased by the retrovirus with ultrasound exposure in these cells. Adjunctive ultrasound exposure was associated with enhanced retrovirus‐mediated transgene expression in vitro. Ultrasound associated local gene therapy has potential for not only plasmid‐DNA‐, but also retrovirus‐mediated gene transfer. 相似文献
106.
Abstract Several biotinylated synthetic oligodeoxyribonucleotides have been selectively prepared by a simple and efficient chemical method. This procedure allows the specific and covalent coupling of one biotinmoiety to any 5′-kinased unprotected oliqodeoxyribonucleotide through an aminoalkylphosphoramide linker arm. The reactions are performed in aqueous solutions on unprotected oligonucleotides and proceed cleanly with good yields. This method is insensitive of the length of the polynucleotide, of the nucleotide sequence and of the nature of the 5′-terminal nucleotide. 相似文献
107.
Etsuko Kawashima Yasuhiro Nakanishi Yusuke Terui Hideyuki Tomitori Keiko Kashiwagi Yusuke Ohba 《Nucleosides, nucleotides & nucleic acids》2013,32(4):196-205
Pyrrole polyamide-2′-deoxyguanosine 5′-phosphate hybrid (Hybrid 4) was synthesized and evaluated in terms of the inhibition of mouse mammary carcinoma FM3A cell growth. Hybrid 4 was found to exhibit dose-dependent inhibition of cell growth. 相似文献
108.
Takayuki Abe Toshihmi Hatta Kazuyuki Takai Hideki Nakashima Tomoyuki Yokota Hiroshi Takaku 《Nucleosides, nucleotides & nucleic acids》2013,32(1-3):471-478
Abstract We have demonstrated that antisense phosphorothioate oligonucleotides (S-ODNs) inhibit influenza virus A replication in MDCK cells. Phosphorothioate and liposomally encapsulated oligonucleotides with two target sites (PB1 and PB2) were synthesized and tested for virus-induced cytopathogenicity effects by a MTT assay using MDCK cells. The liposomally encapsulated S-ODNs complementary to the sites of the PB2-AUG initiation codon showed highly inhibitory effects. On the other hand, the inhibitory effect of the liposomally encapsulated S-ODNs targeted to PB1 was considerably decreased in comparison with the PB2 target sites. The liposomally encapsulated oligonucleotides exhibited higher inhibitory activity than the free oligonucleotides. The activities of the modified oligonucleotides are effectively enhanced by using the liposomal carrier. 相似文献
109.
Sachiko N. Isobe Hideki Hirakawa Shusei Sato Fumi Maeda Masami Ishikawa Toshiki Mori Yuko Yamamoto Kenta Shirasawa Mitsuhiro Kimura Masanobu Fukami Fujio Hashizume Tomoko Tsuji Shigemi Sasamoto Midori Kato Keiko Nanri Hisano Tsuruoka Chiharu Minami Chika Takahashi Tsuyuko Wada Akiko Ono Kumiko Kawashima Naomi Nakazaki Yoshie Kishida Mitsuyo Kohara Shinobu Nakayama Manabu Yamada Tsunakazu Fujishiro Akiko Watanabe Satoshi Tabata 《DNA research》2013,20(1):79-92
110.
Kenta Shirasawa Hiroyuki Fukuoka Hiroshi Matsunaga Yuhko Kobayashi Issei Kobayashi Hideki Hirakawa Sachiko Isobe Satoshi Tabata 《DNA research》2013,20(6):593-603
With the aim of understanding relationship between genetic and phenotypic variations in cultivated tomato, single nucleotide polymorphism (SNP) markers covering the whole genome of cultivated tomato were developed and genome-wide association studies (GWAS) were performed. The whole genomes of six tomato lines were sequenced with the ABI-5500xl SOLiD sequencer. Sequence reads covering ∼13.7× of the genome for each line were obtained, and mapped onto tomato reference genomes (SL2.40) to detect ∼1.5 million SNP candidates. Of the identified SNPs, 1.5% were considered to confer gene functions. In the subsequent Illumina GoldenGate assay for 1536 SNPs, 1293 SNPs were successfully genotyped, and 1248 showed polymorphisms among 663 tomato accessions. The whole-genome linkage disequilibrium (LD) analysis detected highly biased LD decays between euchromatic (58 kb) and heterochromatic regions (13.8 Mb). Subsequent GWAS identified SNPs that were significantly associated with agronomical traits, with SNP loci located near genes that were previously reported as candidates for these traits. This study demonstrates that attractive loci can be identified by performing GWAS with a large number of SNPs obtained from re-sequencing analysis. 相似文献