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1.
Gene delivery systems are designed to control the location of administered therapeutic genes within a patient's body. Successful in vivo gene transfer may require (i) the condensation of plasmid and its protection from nuclease degradation, (ii) cellular interaction and internalization of condensed plasmid, (iii) escape of plasmid from endosomes (if endocytosis is involved), and (iv) plasmid entry into cell nuclei. Expression plasmids encoding a therapeutic protein can be, for instance, complexed with cationic liposomes or micelles in order to achieve effective in vivo gene transfer. A thorough knowledge of pharmaceutics and drug delivery, bio-engineering, as well as cell and molecular biology is required to design optimal systems for gene therapy. This mini-review provides a critical discussion on cationic lipid-based gene delivery systems and their possible uses as pharmaceuticals. 相似文献
2.
Meike Lippelt Theresa Sanabria de Isele Manfred Kist 《Clinical microbiology and infection》1997,3(2):229-235
Objective: To carry out an epidemiologic evaluation of Salmonella enterica subsp. enterica serovar Enteritidis outbreaks in households and small communities by means of rRNA gene restriction pattern analysis (ribotyping).
Method: One hundred Enteritidis isolates dating from 1989 to 1994 which could be allocated epidemiologically to different sources or to small community outbreaks were investigated with ribotyping, a fingerprinting method in which bacterial DNA is hybridized with the biotin-labeled plasmid pKK 3535 containing a ribosomal RNA operon of Escherichia coli to determine the ribosomal RNA gene restriction patterns.
Results: Four different ribotyping patterns were found with the restriction endonuclease Sma I and nine with Sph I. Ribotypes of isolates which could be allocated epidemiologically to a common source usually corresponded. Almost 60% of the Enteritidis infections had the ribotyping pattern Sph I-A. In contrast, this pattern was not found in any of the five Enteritidis strains isolated in 1989. The suspicion that Enteritidis phage type 4 infections are caused by consumption of insufficiently heated eggs is supported by the fact that the ribotyping pattern Sph 1-A was found in isolates from eggs and from human specimens.
Conclusions: As patterns Sph I-A and Sma I-J appeared in 58% and 75% of the isolates, respectively, ribotyping cannot be used for the differentiation between various outbreaks with these two patterns. In cases where the Enteritidis strains showed less frequent patterns, ribotyping seems to be a practical tool for the identification of infection chains. In addition newly appearing ribotyping patterns can give information about the epidemiologic development of Enteritidis infection. 相似文献
Method: One hundred Enteritidis isolates dating from 1989 to 1994 which could be allocated epidemiologically to different sources or to small community outbreaks were investigated with ribotyping, a fingerprinting method in which bacterial DNA is hybridized with the biotin-labeled plasmid pKK 3535 containing a ribosomal RNA operon of Escherichia coli to determine the ribosomal RNA gene restriction patterns.
Results: Four different ribotyping patterns were found with the restriction endonuclease Sma I and nine with Sph I. Ribotypes of isolates which could be allocated epidemiologically to a common source usually corresponded. Almost 60% of the Enteritidis infections had the ribotyping pattern Sph I-A. In contrast, this pattern was not found in any of the five Enteritidis strains isolated in 1989. The suspicion that Enteritidis phage type 4 infections are caused by consumption of insufficiently heated eggs is supported by the fact that the ribotyping pattern Sph 1-A was found in isolates from eggs and from human specimens.
Conclusions: As patterns Sph I-A and Sma I-J appeared in 58% and 75% of the isolates, respectively, ribotyping cannot be used for the differentiation between various outbreaks with these two patterns. In cases where the Enteritidis strains showed less frequent patterns, ribotyping seems to be a practical tool for the identification of infection chains. In addition newly appearing ribotyping patterns can give information about the epidemiologic development of Enteritidis infection. 相似文献
3.
为探讨医院内细菌感染的流行病学特点,作者借助临床分离的64株肺为克雷伯菌,45株阴沟肠杆菌和63株醋酸钙不动杆菌,进行质粒图谱分3种细菌分别有58株,35株和41株含有质粒,且分别构成46个,21个和23个质粒图谱型。结果表明:质粒分析为查明医院内细菌感染源和感染途径提供了较为直接,准确的客观依据,同时也看到了质粒分析的局限性。 相似文献
4.
用限制性内切酶EcoRI从pKS(-)HTH_1切下全长为1.9 kb的人酪氨酸羟化酶基因,在T_4DNA连接酶的作用下连接在真核表达载体pCDNA_3的EcoR Ⅰ位点,构建成重组质粒pcD-NA_3HTH_1,该质粒转染COS-7细胞,免疫荧光组织化学染色证实酪氨酸羟化酶在其中的表达。 相似文献
5.
A study of the value of electrophoretic and other techniques for typing Acinetobacter calcoaceticus 总被引:1,自引:0,他引:1
Forty-four isolates of Acinetobacter calcoaceticus var anitratus collected during hospital outbreaks were studied using polyacrylamide gel electrophoresis (PAGE), plasmid analysis, antibiograms and biochemical tests to determine their degree of similarity. Reproducibility tests were also carried out on the PAGE and biochemical techniques to determine their validity when used to compare bacteria of the same type isolated intermittently. PAGE data was analysed densitometrically and isolates compared using a similarity matrix. All methods were able to subdivide the isolates, but results did not always correlate well between methods. Reproducibility data indicated that careful attention to technique is required when organisms are examined by PAGE sequentially. Results suggest that no single biotyping technique is likely to be adequate and that electrophoretic, biochemical and antibiogram data may complement one another and other epidemiological data in the typing of these organisms. 相似文献
6.
N. I. Buyanova V. P. Shipkov A. P. Pekhov 《Bulletin of experimental biology and medicine》1991,112(5):1646-1649
Department of Biology and General Genetics, Patrice Lumumba Peoples' Friendship University. (Presented by Academician of the Academy of Medical Sciences of the USSR T. T. Berezov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 11, pp. 532–534, November, 1991. 相似文献
7.
目的 克隆人野生型parkin基因并构建真核表达载体pCDNA3.1—parkin,将重组质粒转染PC12细胞获得高表达人野生型parkin基因的PC12细胞克隆。方法 从胎脑组织中提取总RNA,用RT—PCR方法获得人野生型parkin基因的全长cDNA,插入pCR2.1—TA克隆载体中进行序列测定,测序正确后将其亚克隆至表达载体pCD—NA3.1,利用脂质体将重组质粒转染PC12细胞,经G418筛选获得抗性细胞克隆,采用RT—PCR和Western Blot方法鉴定人野生型parkin基因在PC12细胞中的过表达。结果 经限制性内切酶酶切图谱分析和DNA序列测定证实目的基因已插入重组质粒,RT—PCR和Western Blot证明经G418筛选得到的转基因PC12细胞克隆中存在人野生型parkin基因的表达。结论 成功构建了人野生型parkin基因的真核表达载体,获得了稳定表达人野生型parkin基因的PC12细胞克隆,为进一步研究parkin的生物学功能以及parkin在帕金森病发病机制中的作用奠定了良好的基础。 相似文献
8.
9.
本实验将赖型钩端螺旋体(简称钩体)DNA基因库的克隆pCX7制备成 ̄(32)P-重组DNA探针,对8个不同血清群的17株问号状钩体、双曲钩体PatocⅠ株以及细螺旋体3055株DNA进行打点杂交;同时用15种DNA片断进行限制性内切酶谱分析。结果表明,该重组DNA具有问号状钩体种(Species)特异性,但与不同问号状钩体之间的同源性程度有差别;限制性内切酶谱分析发现pCX7重组DNA片段长约1.7kb,具有1个Bg1Ⅱ识别位点和3个BstB1识别位点。 相似文献
10.
Expression of human immunodeficiency virus type 1gag gene using genetically engineered herpes simplex virus type 1 recombinants 总被引:1,自引:0,他引:1
Angela Rösen-Wolff Karl Raab Lothar Zöller Gholamreza Darai Josef Eberle Friedrich Deinhardt 《Virus genes》1990,4(4):325-337
Infectious herpes simplex virus type 1 (HSV-1) recombinants were constructed by inserting the cDNA sequence of the human immunodeficiency virus type 1 (HIV-1)gag gene (from nucleotide position 675 [SacI] to 3859 [Asp 718] of the cDNA sequences of HIV-1 strain BH-10) within the DNA sequences of theBamHI DNA fragment B of the genome of an apathogenic HSV-1 strain HFEM. This HSV-1 strain possesses a 4.1-kbp deletion within theBamHI DNA fragment B between 0.762 and 0.789 map units of the viral genome, which allows the insertion of at least 4 kbp of foreign genetic material into this particular region. The DNA sequences of the immediate early promoter (IE4) of HSV-1 that were inserted upstream from thegag gene were used as a promoter. The screening of 205 virus stocks derived from individual plaques revealed that 46 recombinant viruses harbor HIV-1gag-specific DNA sequences. However, it was found that only six of the recombinant viruses are able to express thegag gene product of HIV-1. This indicates that the ratio of the positive recombination events is about 2.9%. 相似文献