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81.
Maki Ohtani Jun-ichi Hikima Tae-Sung Jung Hidehiro Kondo Ikuo Hirono Haruko Takeyama Takashi Aoki 《Fish & shellfish immunology》2013,34(2):724-728
Phage display libraries are used to screen for nucleotide sequences that encode immunoglobulin variable (V) regions that are specific for a target antigen. We previously constructed an immunoglobulin new antigen receptor (IgNAR) phage display library. Here we used this library to obtain an IgNAR V region that is specific for viral hemorrhagic septicemia virus (VHSV). A phage clone (clone 653) was found to be specific for VHSV by the biopanning method. The V region of clone 653 was used to construct a 6 × His tagged recombinant IgNAR-653 V protein (rIgNAR-653) using the Escherichia coli pET system. The rIgNAR-653 protein bound specifically to VHSV, confirming its activity. 相似文献
82.
Takamichi Izumi Makoto Kondo Takuya Takahashi Nao Fujieda Atsushi Kondo Naohisa Tamura Tomohiro Murakawa Jun Nakajima Hirokazu Matsushita Kazuhiro Kakimi 《Cytotherapy》2013,15(4):481-491
Background aimsAdoptive immunotherapy is emerging as a potent anti-tumor treatment modality; Vγ9Vδ2 T cells may represent appropriate agents for such cancer immunotherapy. To improve the currently limited success of Vγ9Vδ2 T-cell–based immunotherapy, we examined the in vivo dynamics of these adoptively-transferred cells and hypothesized that interleukin (IL)-15 is the potential factor for Vγ9δ2 T cell in vivo survival.MethodsWe conducted a clinical trial of adoptive Vγ9Vδ2 T-cell transfer therapy in six colorectal cancer patients who received pulmonary metastasectomy. Patients' peripheral blood mononuclear cells were stimulated with zoledronate (5 μmol/L) and IL-2 (1000 IU/mL) for 14 d. Harvested cells, mostly Vγ9Vδ2 T cells, were given intravenously weekly without additional IL-2 eight times in total. The frequency, phenotype and common γ-chain cytokine receptor expression of Vγ9Vδ2 T cells in peripheral blood was monitored by flow cytometry at each time point during treatment and 4 and 12 weeks after the last administration.ResultsAdoptively transferred Vγ9Vδ2 T cells expanded well without exogenous IL-2 administration or lymphodepleting preconditioning. They maintained effector functions in terms of interferon-γ secretion and prompt release of cytotoxic granules in response to PMA/ionomycin or isopentenyl pyrophosphate–positive cells. Because they are IL-2Rα?IL-7Rα?IL-15Rα?IL-2Rβ+γc+, it is likely that IL-2 or IL-15 is required for their maintenance.ConclusionsThe persistence of large numbers of functionally active adoptively transferred Vγ9Vδ2 T cells in the absence of exogenous IL-2 implies that an endogenous factor, such as IL-15 transpresentation, is adequate to support these cells in vivo. 相似文献
83.
Carboxy PROXYL is a useful extracellular paramagnetic contrast reagent in electron spin resonance (ESR) and magnetic resonance imaging (MRI). Active transfer of the probe was investigated using an in situ liver model in rats. Carboxy PROXYL, a nitroxyl spin probe, was perfused into in situ liver perfusion system from Wistar rats. Concentration of nitroxyl form of the spin probe in effluent increased gradually after introducing perfusate with the spin probe and reached a plateau. The disappearance of Carboxy PROXYL from the perfusate was 40%, which could not be explained with its partition coefficient. Administration of non-selective inhibitors of organic anion transporters, p-aminohippuric acid and penicillin G, inhibited competitively and in a dose dependent manner the transfer of Carboxy PROXYL into rat liver in situ, resulting in increases of Carboxy PROXYL in the effluent. The results demonstrate that there is an active transfer system of an ESR contrast reagent into in situ rat liver through organic anion transporters. 相似文献
84.
85.
Nami Masubuchi Yuichi Shidoh Shunzo Kondo Jun Takatoh Kazunori Hanaoka 《Experimental Animals》2013,62(3):211-217
Duchenne muscular dystrophy (DMD) is an X-linked recessive progressive muscle
degenerative disorder that causes dilated cardiomyopathy in the second decade of life in
affected males. Dystrophin, the gene responsible for DMD, encodes
full-length dystrophin and various short dystrophin isoforms. In the mouse heart,
full-length dystrophin Dp427 and a short dystrophin isoform, Dp71, are expressed. In this
study, we intended to clarify the functions of these dystrophin isoforms in DMD-related
cardiomyopathy. We used two strains of mice: mdx mice, in which Dp427 was
absent but Dp71 was present, and DMD-null mice, in which both were
absent. By immunohistochemical staining and density-gradient centrifugation, we found that
Dp427 was located in the cardiac sarcolemma and also at the T-tubules, whereas Dp71 was
specifically located at the T-tubules. In order to determine whether T tubule-associated
Dp71 was involved in DMD-related cardiac disruption, we compared the cardiac phenotypes
between DMD-null mice and mdx mice. Both
DMD-null mice and mdx mice exhibited severe necrosis,
which was followed by fibrosis in cardiac muscle. However, we could not detect a
significant difference in myocardial fibrosis between mdx mice and
DMD-null mice. Based on the present results, we have shown that cardiac
myopathy is caused predominantly by a deficiency of full-length dystrophin Dp427. 相似文献
86.
Y. Kondo K. Sho M. A. Majid F. Okajima 《Nucleosides, nucleotides & nucleic acids》2013,32(5):1217-1218
Abstract In thyroid cells, a PI-agonist, via G1 like protein, enhanced a TSH-induced I?-efflux by intensifying a TSH-dependent inositol polyphosphate production followed by a Ca2+ mobilization, but diminished a TSH-dependent DNA synthesis by attenuating a TSH-dependent cAMP accumulation. 相似文献
87.
Akiko Komatsu Hideki Kondo Masayuki Sato Atsushi Kurahashi Kozo Nishibori Nobuhiro Suzuki Fumihiro Fujimori 《Mycoscience》2019,60(4):211-220
Grifola frondosa (Maitake mushroom) is an important cultivated mushroom due to its medicinal and nutrient values. In this study, we isolated and characterized a novel partitivirus (named Grifola frondosa partitivirus 1, GfPV1) infecting a standard G. frondosa strain Gf-N2. This virus has a two-segmented dsRNA genome (dsRNA1 and dsRNA2) with nucleotide lengths of 2.3 and 2.2 kbp, respectively. The coding strand of dsRNA1 and dsRNA2 segments carries single open reading frame encoding RNA-dependent RNA polymerase (RdRp) and a coat protein (CP), respectively. BLAST searches and phylogenetic analyses showed that GfPV1 is most closely related to a betapartitivirus, Lentinula edodes partitivirus 1 (RdRp <70% and CP <60% amino acid sequence identities), but the sequence divergence suggests that GfPV1 is classifiable as a new member of the genus Betapartitivirus, family Partitiviridae. The presence of GfPV1 does not affect colony morphology and fruiting body development of G. frondosa. This is the first report investigating the effects of a mycovirus infection on the colony morphology and fruiting body development of G. frondosa. Interestingly, GfPV1 accumulations markedly decreased along with the fruiting body maturation stages, suggesting the inhibition of virus multiplication during sexual phase of the G. frondosa life cycle. 相似文献
88.
Vertebral bodies of teleost fish are formed by the sclerotomal bone covering the chordacentrum. The internal part of the sclerotomal bone is composed of an amphicoelous hourglass shaped autocentrum, which is common in most fish species. In contrast, the external shape of the sclerotomal bone varies extensively among species. There are multiple hypotheses regarding the composition and formation of the external structure. However, as they are based on studies of few extant or extinct species, their applicability to other species remains to be clarified. To understand the morphology, formation, and composition of vertebral bodies in teleosts, we performed a comparative analysis using micro-CT scans of 32 species from 10 orders of Teleostei and investigated the detailed morphology of the sclerotomal bone, especially its plate-like ridge and trabeculae. We discovered two structural characteristics that are shared among most of the examined species. One was the sheet-like trabeculae that extend radially from the center of the vertebral body with a constant thickness. The other was the presence of hollow spaces on the internal parts of the lateral ridge and trabeculae. The combination of different arrangements of sheet-like trabeculae and internal hollow spaces formed different shapes of the lateral structure of the vertebral body. The properties of these two characteristics suggest that the external part of the sclerotomal bone grows outward by deposition at the bone tip, and that, concurrently, bone absorption occurs in the internal part of the sclerotomal bone. The vertebral arches were also formed by the sheet-like trabeculae, indicating that both, the vertebral body and the arches, are formed by the same component. The micro-CT scanning data were uploaded to a public database so they can be used for future studies on fish vertebrae. 相似文献
89.
Satoshi Suzuki Hisao Hayashi Kenji Takagi Takaaki Kondo Kenzo Takagi Jun Ueyama Shinya Wakusawa 《Biochimica et Biophysica Acta (BBA)/General Subjects》2006
The MDR3 protein is a transporter of phosphatidylcholine on the canalicular membrane of human hepatocytes. Previously we showed that the expression of MDR3 mRNA was down-regulated by phorbol 12-myristate 13-acetate (PMA) in human Chang liver cells. In the present study, to elucidate the isoform of protein kinase C (PKC), which influences the level of MDR3 protein, we investigated the effects of PKC-specific inhibitors and antisense oligonucleotides. The level of protein decreased around 50% after treatment for 3–5 days using the dosage of PMA effective against the mRNA expression. The half-life of the MDR3 protein was estimated to be about 5 days. This decrease was antagonized by GF109203X, a non-selective inhibitor of PKCs, and Gö6976, a selective inhibitor for PKCα/β. These inhibitors also suppressed the reduction in MDR3 protein. To specify the isoform of PKC, the cells were treated with antisense oligonucleotide of PKCα or PKCβ. The suppressive effects on MDR3 mRNA of PMA were attenuated in antisense PKCβ-treated cells, but those in antisense PKCα-treated cells were not attenuated. These suggested that PKCβ plays a regulatory role in the expression of MDR3. 相似文献
90.