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61.
Molecular cloning and sequence analysis of full-length cDNAs encoding new group of Cyn d 1 isoallergens 总被引:3,自引:0,他引:3
BACKGROUND: Cyn d 1, the major allergen of Bermuda grass pollen, contains some acidic/basic isoforms. The N-terminal amino acid sequences of some acidic Cyn d 1 isoforms were found to be different from those of Cyn d 1 cDNA clones identified previously. METHODS: A predicted 17-meric oligonucleotide probe was designed to fish the unidentified isoallergen cDNAs out of BGP cDNA library. The reactive clones were isolated and verified by sequencing. Two of them were expressed in the yeast Pichia pastoris to obtain recombinant Cyn d 1 proteins. RESULTS: All four cDNA clones encode the full-length Cyn d 1 with mature proteins of 244 amino acid residues. A 97-99% identity was found among the deduced amino acids of these four clones while an 86% identity was elicited between the four clones and the ones previously identified. The predicted isoelectric focusing (pI) values of the newly identified Cyn d 1s are acidic while pIs of the previously identified Cyn d 1s are basic. The two recombinant acidic Cyn d 1 proteins possess the epitopes recognized by mouse and rabbit polyclonal anti-Cyn d 1 antibodies, and have human IgE-binding capacity as revealed by immunodot assay. CONCLUSIONS: The present study identified full-length cDNAs encoding new isoallergens of Cyn d 1, and separated Cyn d 1 gene into an acidic group and a basic group. 相似文献
62.
Bin Yu Youming Ding Xiaofeng Liao Changhua Wang Bin Wang Xiaoyan Chen 《Pathology, research and practice》2019,215(5):939-945
Background
TONSL has been suggested to function as an oncogene in lung, esophageal and cervical cancer. This study was aimed to identify the expression of TONSL and its role in hepatocellular carcinoma (HCC).Methods
By data mining in the Cancer Genome Atlas (TCGA) and Human Protein Atlas (HPA) databases, the expression profile of TONSL, its clinical significance, the potential mechanisms of its dysregulation and its underlying biological function in HCC were investigated.Results
TONSL was significantly upregulated in HCC tissues relative to normal liver tissues (P?<?0.05). High TONSL expression was significantly correlated with advanced TNM stage, poorly differentiated tumors, vascular invasion, elevated serum alpha-fetoprotein expression and a worse prognosis (all P?<?0.05). Multivariate analysis further confirmed that TONSL overexpression was an independent risk factor for poor overall survival (OS) and recurrence-free survival (RFS) in HCC (all P?<?0.05). Additionally, 16% of HCC cases (n?=?370) had TONSL DNA amplification. The total methylation level of TONSL was moderately and negatively correlated with its mRNA expression (P?<?0.05). TONSL was predictively targeted by miR-133b, which was downregulated in HCC and negatively related to TONSL mRNA expression (all P?<?0.05). Kaplan-Meier analyses demonstrated that low miR-133b expression was significantly associated with poor OS and RFS (all P?<?0.05). Moreover, gene set enrichment analysis revealed that cases with TONSL overexpression were enriched in cell cycle regulation pathways (all P?<?0.05).Conclusions
TONSL holds promise for serving as a prognostic biomarker for HCC. DNA amplification, hypomethylation and miR-133b downregulation could be the mechanisms associated with TONSL upregulation in HCC. TONSL might function as an oncogene via cell cycle regulation pathways in HCC. 相似文献63.
Difference in cell binding patterns of two monoclonal antibodies recognizing distinct epitopes on a human melanoma-associated oncofetal antigen 总被引:1,自引:0,他引:1
Two monoclonal antibodies (MAbs), 140.240 and 96.5, generated independently in different laboratories, have been shown to detect the target structures of 87,000 (gp87) and 97,000 (p97) glycoproteins, respectively, both strongly expressed by melanoma cells and fetal small intestine. To determine whether MAb 140.240 and MAb 696.5 recognized a same target structure, they were tested in immunoprecipitation/SDS-PAGE using NP-40 lysates of melanoma cells labelled with [35S]methionine for 18 hr. Both antibodies precipitated a single band with Mr = 87,000. Reciprocal immunodepletion studies showed that neither of the two antibodies detected the 87,000 band in the lysate immuno depleted by either antibody, suggesting that these two antibodies recognize the same or extremely similar molecules. Two-dimensional tryptic peptide mapping analysis showed that the two identified molecules shared the same finger-printing pattern. A 40,000 fragment of the 87,000 molecule produced by protease digestion was precipitated by MAb 96.5 but not MAb 140.240, indicating that the epitopes recognized by the two antibodies are localized at discrete sites on the molecule. Serological studies on these two antibodies revealed slightly different binding patterns in the MAb 140.240 exhibited a more melanoma-restricted specificity, while MAb 96.5 had a specificity to melanoma and to some other cell types. The observed difference in epitope specificity may be important in the clinical applications of these antibodies. 相似文献
64.
Yongchol Shin Atsushi Kitayama Tetsuya Koide Daniel A Peiffer Makoto Mochii Arnold Liao Naoto Ueno Ken W Y Cho 《Developmental dynamics》2005,232(2):432-444
To isolate novel genes regulating neural induction, we used a DNA microarray approach. As neural induction is thought to occur by means of the inhibition of bone morphogenetic protein (BMP) signaling, BMP signaling was inhibited in ectodermal cells by overexpression of a dominant-negative receptor. RNAs were isolated from control animal cap explants and from dominant-negative BMP receptor expressing animal caps and subjected to a microarray experiment using newly generated high-density Xenopus DNA microarray chips representing over 17,000 unigenes. We have identified 77 genes that are induced in animal caps after inhibition of BMP signaling, and all of these genes were subjected to whole-mount in situ hybridization analysis. Thirty-two genes showed specific expression in neural tissues. Of the 32, 14 genes have never been linked to neural induction. Two genes that are highly induced by BMP inhibition are inhibitors of Wnt signaling, suggesting that a key step in neural induction is to produce Wnt antagonists to promote anterior neural plate development. Our current analysis also proves that a microarray approach is useful in identifying novel candidate factors involved in neural induction and patterning. 相似文献
65.
66.
At the University of California, Irvine Medical Center, an end-of-life curriculum was implemented in 2000 for an internal medicine residency utilizing a longitudinal approach that allowed residents to follow patients through their entire hospice experience. An elective home hospice rotation was developed for which third-year residents served as primary care physicians for patients at the end of life over a one-year period. Residents were supervised by faculty who were hospice medical directors. They also learned through case vignettes, quarterly meetings, textbook reading, and personal projects. From July 2000 to June 2002, residents demonstrated positive attitudes towards hospice care and recommended the rotation highly (mean 8.86 on a scale of 1-10). The rotation grew in popularity from six initial residents to ten residents the next year, and has since become a mandatory rotation for all senior residents. A 360-degree evaluation uniformly indicated positive resident performance from the hospice team (mean scores 7.56-8.69 on a 1-9 scale), family (mean scores 9.3-9.7 on a 1-10 scale) and faculty (mean scores 7.29-7.72 on a 1-9 scale). Residents were also pleased with the level of teaching (mean 8.86 on a scale of 1-10) and felt that the patient care load was "just right." Their knowledge improved by 8% (p =.0175). In conclusion, a longitudinal hospice rotation was implemented that fulfilled curricular goals without undue burden on the residents or residency program. 相似文献
67.
Different applications of polymerases with and without proofreading activity in single-nucleotide polymorphism analysis 总被引:2,自引:0,他引:2
Zhang J Li K Liao D Pardinas JR Chen L Zhang X 《Laboratory investigation; a journal of technical methods and pathology》2003,83(8):1147-1154
With the completion of the human genome project, single-nucleotide polymorphisms (SNPs) have become the focus of intense study in biomedical research. Polymerase-mediated primer extension has been employed in a variety of SNP assays. However, these SNP assays using polymerase without proofreading function are compromised by their low reliability. Using a newly developed short amplicon harboring restriction enzyme site, EcoR-I, we were able to compare the single-base discrimination abilities of polymerases with and without proofreading function in primer extension in a broad range of annealing temperatures. Thermodynamic analysis demonstrated a striking single-nucleotide discrimination ability of polymerases with proofreading function. Using unmodified 3'-end allele-specific primers, only template-dependent products were generated by polymerase with proofreading activity. This powerful single-base discrimination ability of exo(+) polymerases was further evaluated in primer extension using three types of 3' terminally modified allele-specific primers. As compared with the poor fidelity in primer extension of polymerases lacking 3' exonuclease activity, this study provides convincing evidence that the use of proofreading polymerases in combination with 3'-end modified allele-specific primers can be a powerful new strategy for the development of SNP assays. 相似文献
68.
Autoimmunity in the pathogenesis of cardiomyopathy 总被引:3,自引:0,他引:3
69.
主诉腰痛女性患者骨密度和骨质疏松患病率分析 总被引:3,自引:3,他引:0
目的研究主诉腰痛女性患者腰椎和髋部骨密度(BMD)测量的临床意义及骨质疏松(OP)患病率。方法采用扇形束双能X线骨密度仪(DEXA)测量主诉为腰痛的女性患者的腰椎和髋部BMD,并与对照组的BMD比较,同时比较其相应骨质疏松(OP)患病率。结果临床主诉以腰痛为主的女性病人腰椎及髋部BMD比以关节痛或全身痛为主诉者的低,(P<0.05);主诉腰痛者各骨骼部位的OP检出率明显高于关节痛或全身痛者,且均与正常对照组有显著性差异(P<0.05)。结论骨质疏松是女性腰痛病人就诊的主要原因之一,腰痛患者OP患病率较高;腰椎BMD测定是诊断骨质疏松症的重要手段。 相似文献
70.
梯度胶电泳法测定冠心病患者血浆小密低密度脂蛋白 总被引:1,自引:0,他引:1
目的 建立实用的小密低密度脂蛋白(sdLDL)测定方法, 观察冠心病患者血浆中sdLDL和脂类成分的变化及相互关系。方法 采用2%~16%非变性聚丙烯酰胺梯度胶电泳和凝胶成象分析系统, 测定了105名冠心病患者和90名健康对照者血浆sdLDL、 血脂。结果 sdLDL>50%的检出率, 冠心病组为38.1%, 对照组为11.1%(P<0.01);sdLDL增多患者的甘油三酯水平(2.43±1.22) mmol/L明显高于sdLDL正常者(1.46±0.64) mmol/L, P<0.01。结论 sdLDL与冠心病发生发展有关, 且与甘油三酯关系密切;sdLDL测定可作为预测冠心病危险性的指标之一。 相似文献