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991.
The nucleotide sequence of the 3' terminal 2022 nucleotides (nt) of tobacco ringspot virus (TobRV) RNA 2 has been determined. Protein microsequence analysis of the amino-terminal residues of purified capsid protein localized the capsid protein gene between nt 2014 and 583 (from the 3' terminus) of this sequence. The proteolytic cleavage site that is processed to liberate the capsid protein from the RNA 2-encoded polyprotein was identified as Cys-Ala. The predicted translation product from the gene is a 477 amino acid long polypeptide with a calculated MW of 53 kDa. The gene was modified at the 5' end to facilitate sub-cloning, and to provide it with a methionine initiation codon. The modified gene was sub-cloned, transcribed in vitro and expressed in a rabbit reticulocyte lysate translation system, where it directed the synthesis of a 53 kDa polypeptide. Garnier-Osguthorpe-Robson analyses of the secondary structure of the capsid protein predicted the presence of three beta sheet domains, which suggests that this nepovirus capsid may be structurally analogous to those of the como- and picornaviruses. These and other results from computer analyses of the nucleic acid and amino acid sequences, and comparisons with the capsid proteins of nepoviruses and other related viruses are discussed. 相似文献
992.
An episomal DNA vector (YpJA18), encoding two selectable recombinant yeast genes (TRP1, URA3), was constructed to assess the fidelity of DNA repair in haploid repair-competent (RAD) wild-type yeast and several radiation-sensitive mutants. Either a DNA double-strand break (DSB) or a double-strand gap of 169 bp (DSG) was introduced by restriction enzymes in-vitro within the coding sequence of the URA3 gene of this vector. To eliminate transfer artefacts, selection was first applied for the undamaged TRP1 gene followed by counter selection for URA3 gene activity, which indicated correct repair of the DSB and DSG. Correct repair of the damaged URA3 gene was found to be about 90% in RAD cells (normalized for the expression of undamaged URA3 in TRP
+ transformants). Plasmids isolated from the transformants (URA
+
TRP
+) carry both unique sites (ApaI and NcoI) within the URA3 gene indicating the precise restitution of the 169-bp gap. An excision-repair-defective rad4-4 mutant repaired these lesions as correctly as RAD cells, whereas the mutants rad50-1, rad51-1 and rad54-1, proven to be defective in DSB repair and mitotic recombination, showed less than 5% correct repair of such lesions. In contrast, a representative of the RAD6 epistasis group of genes, the rev2-1 mutant which is sensitive towards UV and ionizing radiation, had a significantly reduced ability (about 20%) for the correct repair of both DSBs and DSGs. 相似文献
993.
Takashi Sekine Keiko Fukutani Tomiko Motegi Hiroshi Hayakawa Takashi Tamura Shigeo Nagafuchi Yutaka Nakahori Yasuo Nakagome 《Journal of human genetics》1992,37(2):157-162
Summary Results of DNA study on two patients of gonadal dysgenesis with a 45,X/46,X,Ynf (non-fluorescent Y chromosome) karyotype are described. In one patient who developed gonadoblastoma, all 12 loci on the non-fluorescent part of Yq were detected. Another patient did not have gonadoblastoma at 20 years, and only the proximal 6 loci out of 12 were detected. 相似文献
994.
Functional mapping of the activity of the R region in the human T-cell leukemia virus type I long terminal repeat to increase gene expression 总被引:3,自引:0,他引:3
We previously demonstrated the activity of the R fragment in the long terminal repeat of human T-cell leukemia virus type I for elevation of the level of gene expression. In this study, the fragment was deleted with BAL31 nuclease to determine its functional domain. Series of the shortened R fragments were linked to the simian virus 40 promoter unit, which regulated expression of a reporter gene. Examination with the R fragments deleted from the 5 and 3 ends showed that borders of the functional domain were mapped within nucleotide positions 458 to 473 for the 5 end and nucleotide positions 559 to 594 for the 3 end, respectively. Thus we conclude that a 136-base-pair fragment corresponding to the second half of the R region was sufficient to allow elevation of the level of gene expression. 相似文献
995.
996.
997.
颅内海绵状血管瘤 CCM1基因第12外显子及其5′端内含子新突变位点 总被引:2,自引:0,他引:2
目的 探讨 CCM1基因突变在中国人颅内海绵状血管瘤 ( intracranial cavernous angiomas,ICCA)发病中所起的作用。方法 收集我院神经外科 2 0 0 2年 6月~ 2 0 0 3年 2月收治并经手术病理证实的2 1例 ICCA患者及 15名正常健康对照者 ,从外周静脉血中提取 DNA,PCR法扩增 CCM1基因第 12外显子及其两侧部分内含子序列 ,应用 DNA直接测序技术对扩增产物进行检测。结果 5例患者中检测出 3处 CCM1基因突变 ,均为首次发现。其中 ,5例患者中均存在 1172 C→ T的错义突变 ,使编码 KRIT1蛋白391位的氨基酸由丝氨酸变成苯丙氨酸。另有 1例患者存在 116 0 A→ C的错义突变 ,使编码 KRIT1蛋白387位氨基酸的谷氨酰胺变成脯氨酸。另一个突变发生在第 12外显子 5′端内含子区域 ,5例患者中有 4例第 4个碱基 C被 T取代。对照组检测结果无异常。结论 中国 ICCA患者存在 CCM1基因第 12外显子的突变 ,并与 ICCA的发病有关 相似文献
998.
为了解异基因骨髓移植治疗某些遗传及恶性血液性疾病的植活监测指标,用聚合酶链式反应(PCR)扩增具有高度多态性的小卫星DNA33.1、33.6位点,联合地高辛标记的位点特异性寡核苷酸探针杂交的方法,以小卫星DNA指纹图为特异性遗传标志,对6例异基因骨髓移植,1例脐血移植进行植活指标监测,6例获得供者源性细胞植活证据。结果显示:小卫星DNA指纹图个体特异性强,可作为监测异基因造血干细胞移植植活的可靠格标,尤其是同血型,同性别,HLA配型表型全相合的移植;该方法程序简单,操作方便,灵敏度高,可达0.5%,最早获得植入证据的时间是+15天,仅需模板DNA100ng;如果扩增位点增加,可提高个体识别能力。 相似文献
999.
采用聚合酶链反应(PCR)技术,对42例肝活切组织石蜡切片中乙型肝炎病毒(HBV)DNA进行检测,并与乙肝表面抗原(HBsAg)的免疫组织化学及血清学检测进行比较,HBV-PCR阳性率为73.8%,高于组织及血清HBsAg阳性率(分别为59.5%和50.0%)。3例病理形态呈肝炎改变,而血清HBsAg(─)的肝组织中有2例检出HBV-DNA,提示PCR的高度敏感性和准确性。83.3%的门脉性肝硬变和87.5%的肝细胞癌组织中HBV-PCR呈阳性,进一步证实了上述两病与HBV的关系密切。我们还发现肝细胞淤胆患者HBV感染率较高,HBV-DNA及组织HBsAg阳性比例各为6/9和4/8。 相似文献
1000.
Two models have been considered to account for the differentiation of γδ and αβ T cells from a common hematopoietic progenitor cell. In one model, progenitor cells commit to a lineage before T cell receptor (TCR) rearrangement occurs. In the other model, progenitor cells first undergo rearrangement of TCRγ, δ, or both genes, and cells that succeed in generating a functional receptor commit to the γδ lineage, while those that do not proceed to attempt complete β and subsequently α gene rearrangements. A prediction of the latter model is that TCRγ rearrangements present in αβ T cells will be nonproductive. We tested this hypothesis by examining Vγ2-Jγ1Cγ1 rearrangements, which are commonly found in αβ T cells. The results indicate that Vγ2-Jγ1Cγ1 rearrangements in purified αβ T cell populations are almost all nonproductive. The low frequency of productive rearrangements of Vγ2 in αβ T cells is apparently not due to a property of the rearrangement machinery, because a transgenic rearrangement substrate, in which the Vγ2 gene harbored a frame-shift mutation that prevents expression at the protein level, was often rearranged in a productive configuration in αβ T cells. The results suggest that progenitor cells which undergo productive rearrangement of their endogenous Vγ2 gene are selectively excluded from the αβ T cell lineage. 相似文献