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151.
Signal transduction pathways in guinea pig sperm 总被引:2,自引:0,他引:2
Trifluoperazine (TFP), the antagonist of calmodulin (CaM). significantly stimulated the capacitation and acrosome reaction of guinea pig spermatozoa at the concentration of 10-100μmol/L, independent of the external Ca2+. Forskolin, dbcAMP and caffeine evidently promoted the occurrence of acrosome reaction of spermatozoa at early capacitation stage (5 h) in nonsynchronous system but not in synchronous system. If the spermatozoa were capacitated for 15 h in synchronous system, the above three drugs significantly stimulated acrosome reaction in a Ca2+-independent manner. Protein kinase C activators, i.e. phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-dibutyrate (PDB) did not influence the occurrence of acrosome reaction of spermatozoa at early capacitation stage, but significantly increased the acrosome reaction rate in capacitated spermatozoa in a Ca2+-independent manner. In contrast. PKC inhibitor staurosporine significantly inhibited the occurrence of acrosome reaction. 相似文献
152.
Giuseppe Comi Corrado Pipan Giuseppe Botta Luca Cocolin Carlo Cantoni Marisa Manzano 《FEMS immunology and medical microbiology》1996,16(1):45-49
Abstract A combined polymerase chain reaction and restriction endonuclease (RE) enzyme assay was developed to discriminate between Campylobacter coli and Campylobacter jejuni . Amplimers of the FlaA gene obtained by PCR were digested with Alu I and Hin fI to distinguish C. coli from C. jejuni . With Alu I digestion C. jejuni -specific bands were observed at 110, 140 and 160 bp and C. coli -specific bands at 293 and 147 bp. C. jejuni -specific bands of 349 and 109 bp were found by Hin fI digestion but Hin fI did not digest the Fla A amplimer of C. coli . This combined technique is fast and easy to perform, and distinguishes the two campylobacters unequivocally. 相似文献
153.
Rapid typing of truffle mycorrhizal roots by PCR amplification of the ribosomal DNA spacers 总被引:4,自引:0,他引:4
DNA analyses were developed to type mycorrhizas of two Tuber species of commercial value (T. melanosporum, T. borchii) and a competitive fungus (Sphaerosporella brunnea) which forms ectomycorrhizas with plants usually considered hosts for truffles. Polymerase chain reaction (PCR) amplification
of DNA isolated from fruitbodies, mycelia, mycorrhizas and leaves of host plants, was performed with a primer pair for an
internal transcribed spacer ITS1-4. ITS amplification followed by restriction fragment length polymorphism (RFLP) analysis
of the amplified products clearly distinguished the two Tuber species at the fruitbody, mycorrhiza and mycelium levels.
Accepted: 6 September 1996 相似文献
154.
改进了一种分析磷酸酶活性的终止酶反应方法.该方法通过在酶反应进行到一定程度时,在反应混合物中加入酶反应终止液(1mol/L NaOH-0.2mol/L EDTA),从而使测定更简捷、精确. 相似文献
155.
156.
一类一级饱和反应系统的极限环 总被引:1,自引:0,他引:1
本文对一类一级饱和反应系统进行定性分析,得到(1)存在一个唯一环和存在两个(内不稳外稳)环的条件。 相似文献
157.
EB病毒胸苷激酶基因的扩增 总被引:1,自引:0,他引:1
EB病毒胸苷激酶基因的扩增陈尚武,陈瑞君,黄迪,朱振宇,马涧泉(中山医科大学生化教研室,广州510089)(中山医科大学肿瘤研究所,广州510060)关键词Epstein-Barr病毒,胸苷激酶,基因,聚合酶链反应鼻咽癌(nasopharyngeal... 相似文献
158.
Balzer Sandrock Karen M. Hudson Douglas E. Williams Michael A. Lieberman 《In vitro cellular & developmental biology. Animal》1996,32(4):225-233
Summary The regulation of megakaryopoeisis by cytokines is not yet well understood. It is possible that autocrine loops are established
during megakaryocyte growth and differentiation, aiding in the maturation of these cells. The CHRF-288-11 human megakaryoblastic
cell line has been examined for cytokine production in growing cells and cells stimulated to differentiate by the addition
of phorbol esters. It has been demonstrated that these cells produce RNA corresponding to the interleukins IL-1α, 1β, 3, 7,
8, and 11, granulocyte-macrophage colony stimulating factor (GM-CSF), stem cell factor (SCF), transforming growth factor-β
(TGF-β), tumor necrosis factor-α (TNF-α), interferon-α (INF-α), and basic fibroblast growth factor (bFGF). Additionaly, RNA
corresponding to the receptors for IL-6, GM-CSF, SCF, INF-α,β, bFGF, and monocyte colony stimulating factor (M-CSF) were also
expressed by the cells. The receptor for TNF-α was detected immunologically. Analysis at the protein level demonstrated that
significant amounts of INF-α, TNF-α, GM-CSF, SCF, IL-1α, and a soluble form of the IL-6 receptor were produced by the cells.
Addition of phorbol esters to CHRF-288-11 cells enhances their megakaryocytic phenotype; such treatment also results in increased
secretion of INF-α, TNF-α, and GM-CSF. These results suggest that potential autocrine loops are established during the differentiation
of CHRF-288-11 cells, which may alter the capability of the cell to differentiate. These findings are similar to those recently
obtained for marrow-derived megakaryocytes (Jiang et al.) suggesting that CHRF-288-11 cells provide a useful model system
for the study of cytokine release during megakaryocyte differentiation. 相似文献
159.
Rui Yan Mark Ottenbreit Bharati Hukku Michael Mally Sharong Chou Joseph Kaplan 《In vitro cellular & developmental biology. Animal》1996,32(10):656-662
Summary Methods for monitoring cell line identification and authentication include species-specific immunofluorescence, isoenzyme
phenotyping, chromosome analysis, and DNA fingerprinting. Most previous studies of DNA fingerprinting of cell lines have used
restriction fragment length polymorphism analysis. In this study, we examined the utility of an alternative and simpler method
of cell line DNA fingerprinting—polymerase chain reaction (PCR) amplification of fragment length polymorphisms. Fourteen human
cell lines previously found by other methods to be either related or disparate were subjected to DNA fingerprinting by PCR
amplification of selected fragment length polymorphism loci. Cell identification patterns by this method were concordant with
those obtained by isoenzyme phenotyping and restriction fragment length polymorphism-DNA fingerprinting, and were reproducible
within and between assays on different DNA extracts of the same cell line. High precision was achieved with electrophoretic
separation of amplified DNA products on high resolution agarose or polyacrylamide gels, and with fragment length polymorphism
(FLP) loci-specific “allelic ladders” to identify individual FLP alleles. Determination of the composite fingerprint of a
cell line at six appropriately chosen fragment length polymorphism loci should achieve a minimum discrimination power of 0.999.
The ability of PCR-based fragment length polymorphism DNA fingerprinting to precisely and accurately identify the alleles
of different human cell lines at multiple polymorphic fragment length polymorphism loci demonstrates the feasibility of developing
a cell line DNA fingerprint reference database as a powerful additional tool for future cell line identification and authentication. 相似文献
160.
将遴选的经适当接尾的12个HLA-DQA1序列特异性寡核苷酸固定在一张滤膜上,用生物素标记的DQA1特异性扩增产物与滤膜上的序列特异性寡核苷酸在四甲基氯化铵杂交体系中杂交,然后经洗膜封膜,杂交信号用非放射性的碱性磷酸酶显色法检测,根据杂交斑点的显示结果分析标本的基因型。采用这种方法初步确定了HLA-DQA1位点8种单倍型等位基因:DQA10101、0102、0103、0201、03011、0401、0501和0601.非放射性反相杂交法可对各种来源的杂合性标本进行HLA-Ⅱ类基因快速分型,并适合在临床器官移植的组织分型配型、疾病易感性研究和法医鉴定等领城中应用。 相似文献