全文获取类型
收费全文 | 269篇 |
免费 | 30篇 |
国内免费 | 2篇 |
专业分类
儿科学 | 8篇 |
妇产科学 | 2篇 |
基础医学 | 74篇 |
口腔科学 | 6篇 |
临床医学 | 24篇 |
内科学 | 60篇 |
皮肤病学 | 5篇 |
神经病学 | 9篇 |
特种医学 | 10篇 |
外科学 | 38篇 |
综合类 | 26篇 |
预防医学 | 8篇 |
药学 | 26篇 |
肿瘤学 | 5篇 |
出版年
2021年 | 3篇 |
2020年 | 1篇 |
2019年 | 2篇 |
2018年 | 5篇 |
2017年 | 2篇 |
2016年 | 4篇 |
2015年 | 10篇 |
2014年 | 10篇 |
2013年 | 11篇 |
2012年 | 7篇 |
2011年 | 8篇 |
2010年 | 11篇 |
2009年 | 23篇 |
2008年 | 20篇 |
2007年 | 5篇 |
2006年 | 8篇 |
2005年 | 6篇 |
2004年 | 5篇 |
2003年 | 4篇 |
2002年 | 11篇 |
2001年 | 8篇 |
2000年 | 12篇 |
1999年 | 10篇 |
1998年 | 9篇 |
1997年 | 11篇 |
1996年 | 15篇 |
1995年 | 5篇 |
1994年 | 10篇 |
1993年 | 17篇 |
1992年 | 10篇 |
1991年 | 6篇 |
1989年 | 5篇 |
1988年 | 3篇 |
1987年 | 2篇 |
1986年 | 2篇 |
1985年 | 2篇 |
1984年 | 1篇 |
1983年 | 1篇 |
1982年 | 1篇 |
1981年 | 3篇 |
1980年 | 1篇 |
1977年 | 2篇 |
1975年 | 2篇 |
1974年 | 1篇 |
1972年 | 2篇 |
1971年 | 2篇 |
1970年 | 2篇 |
排序方式: 共有301条查询结果,搜索用时 31 毫秒
1.
2.
3.
Richard Byers Jamie Roebuck Ebrahim Sakhinia Judith Hoyland 《Diagnostic molecular pathology》2004,13(3):144-150
RNA extraction still relies almost exclusively on the use of fresh or frozen tissue, limiting the number of samples that can be analyzed, and there is a growing need for means of global mRNA analysis of archived formalin-fixed paraffin-embedded tissue (FFPET). Previous reports of RNA extraction and amplification from FFPET are limited and do not enable global cDNA amplification. This study used polyA PCR to generate globally amplified cDNA from RNA extracted from formalin-fixed paraffin-embedded samples. RNA was extracted from nine routinely processed archival FFPET samples (lymph node, nasopharynx, prostate, lung and bone marrow) using an Ambion Paraffin Block RNA Isolation Kit. Global cDNA was generated by polyA RT-PCR and used in GAPDH specific PCR and PCR for CD33, c-myb, and SNF2. PolyA cDNA was reamplified by polyA PCR and the reamplified cDNA also used in GAPDH PCR. RNA was extracted from all nine samples, but was degraded. PolyA RT-PCR generated cDNA from all samples and was positive for GAPDH PCR in seven. PCR for CD33, c-myb, and SNF2 was positive in all samples tested. Following reamplification, the polyA cDNA remained positive for GAPDH by PCR. The results demonstrate the feasibility of globally amplifying RNA isolated from archival FFPET samples using polyA RT-PCR, which generates a renewable cDNA pool that can be probed for any cDNA species and reamplified as necessary. 相似文献
4.
Succinylation of concanavalin A (Con A) reduces its oligomer size while retaining its mitogenicity, and provides a probe of T cell activation. We have observed responses of cytosolic ionized calcium to succinyl Con A in suspensions of Jurkat and rat lymph node (LN) cells, using a fluorimeter, and in single cells settled on glass, using a dual wavelength video imaging system. In the fluorimeter a mitogenic level of succinyl Con A (30 micrograms/ml) produced only a 15-30 nM rise in average cell calcium in the suspended Jurkat or rat cells whereas the use of quantitative video imaging produced asynchronous 250-1000 nM pulses of free calcium in 35% of Jurkat cells and 300-850 nM pulses in 45% of rat LN cells. In Jurkat cells these pulses were sometimes repetitive, giving rise to apparent oscillations. In the fluorimeter 30 micrograms/ml of native Con A (a supra-mitogenic concentration) produced a 300 nM rise in average cell calcium in suspended Jurkat cells, and a 100 nM rise in rat LN cells; when major histocompatibility complex class II-bearing cells were removed the response rose. Mitogenic Con A (3 micrograms/ml) produced a much lower rise in calcium. With video imaging the response seen was greater. Levels greater than 30 micrograms/ml Con A caused 700-5000 nM pulses synchronously in 94% of Jurkat cells and 250-1000 nM pulses in 73% of rat LN cells. At 3 micrograms/ml Con A produced asynchronous 300-1100 nM pulses in 36% of rat LN cells. We conclude that the absence of a calcium signal in the fluorimeter can conceal asynchronous calcium responses in individual cells and that brief asynchronous cytosolic calcium pulses are sufficient for lectin to activate rat T cells. 相似文献
5.
Glycoproteins present in human follicular fluid that inhibit the zona- binding capacity of spermatozoa 总被引:1,自引:0,他引:1
Previous studies have suggested that human follicular fluid contains
factors that reduce the zona-binding capacity of spermatozoa. The present
study provides further evidence of the existence of such factors. Using the
hemizona binding assay (HZA), we have shown that the inhibitory effect of
human follicular fluid on the zona-binding capacity of spermatozoa is
concentration-dependent, an inhibitory effect being detected when the
concentration of human follicular fluid was > or = 10%. A 1%
concentration of human follicular fluid did not possess this inhibitory
activity. Heating human follicular fluid at 56 degrees C for 30 min did not
affect its inhibitory properties; treatment with proteinase-K abolished
such inhibition. Human follicular fluid was fractionated sequentially by
concanavalin-A affinity chromatography, Mono Q ion-exchange chromatography
and Superose-12 gel filtration. The zona binding inhibitory activity
resided in the fraction which bound to the lectin and Mono Q column and
contained molecules with native molecular weights of 32 and 192 kDa. Sodium
dodecyl sulphate-polyacrylamide gel electrophoresis analysis suggested that
the 192 kDa glycoprotein was a tetramer, while the 32 kDa glycoprotein
remained as a single molecular species under denaturing conditions. We
conclude that two glycoproteins were responsible for the zona binding
inhibitory activity of human follicular fluid. The physiological role of
these factors remains unclear.
相似文献
6.
7.
Evidence for cell-specific changes with age in expression of oestrogen receptor (ER) alpha and beta in bone fractures from men and women 总被引:7,自引:0,他引:7
Batra GS Hainey L Freemont AJ Andrew G Saunders PT Hoyland JA Braidman IP 《The Journal of pathology》2003,200(1):65-73
Oestrogen is recognized as important for maintaining bone mass in men and women. Oestrogen receptor (ER) alpha and the recently described ER-beta are both expressed in bone cells, but have different affinities for oestrogen agonists and plant oestrogens, which could be important in developing treatments for bone loss in both men and women. It is unclear, however, which isoform predominates in bone; cell type and age may influence their relative expression. The present study has compared ER-alpha and ER-beta expression in serial sections of human fracture callus from males (n = 19, age range 5-72 years) and females (n = 15, age range 3-86 years) by indirect immunoperoxidase. Fracture callus was used as it can be readily obtained from individuals over a wide age range and contains a variety of bone cells. Antibody specificity was confirmed by western blotting and comparison of immunoreactivity in sections of breast tumour and benign prostate hyperplasia. No gender difference in ER expression was found in bone from individuals less than 40 years old. Proliferative chondrocytes were positive for both isoforms, but few larger hypertrophic cells were immunoreactive. ER-alpha and ER-beta were co-expressed in osteoclasts, suggesting that oestrogen may act directly on these cells. Osteoblasts, osteocytes, and mesenchymal cells also expressed both isoforms. In women over 40 years of age, however, relatively fewer biopsies contained osteocytes positive for ER-alpha and ER-beta. Likewise, the proportions of osteoblasts and mesenchymal cells expressing ER-beta were reduced but ER-alpha remained unaffected. In contrast, in men over 40 years, only the proportion of biopsies containing ER-beta-positive mesenchymal cells was lower. In these older men and women, ER-alpha and ER-beta expression was retained by the small proliferative chondrocytes. These results demonstrate that gender, age, and cell type are important determinants of ER isoform expression in skeletal cells. 相似文献
8.
Current understanding of cellular and molecular events in intervertebral disc degeneration: implications for therapy 总被引:12,自引:0,他引:12
Freemont AJ Watkins A Le Maitre C Jeziorska M Hoyland JA 《The Journal of pathology》2002,196(4):374-379
Until recently, material removed from the intervertebral disc (IVD) at surgery consisted either of 'loose bodies' from the centre of the IVD or discal tissue displaced (prolapsed) into the intervertebral root or spinal canals. This material is best regarded as a by-product of disc degeneration and therefore not representative of the disease process itself. Recent advances in surgical techniques, particularly anterior fusion, in which large segments of the anterior part of the IVD are excised with the anatomical relationships between different components intact, have generated material that can be investigated with modern molecular and cell biological techniques. This is an important area of study because degeneration of the lumbar IVDs is associated, perhaps causally, with low back pain, one of the most common and debilitating conditions in the West. 'Degeneration' carries implications of inevitable progression of wear-and-tear associated conditions. Modern research on human IVD tissue has shown that this is far from the case and that disruption of the micro-anatomy described as degeneration is an active process, regulated by locally produced molecules. The exciting consequence of this observation is the possibility of being able to inhibit or even reverse the processes of degeneration using targeted therapy. 相似文献
9.
MW Lieberman R Barrios G Kala SV Kala ED Lykissa CN Ou 《Environmental health perspectives》1999,107(9):A444-A445
Respond on comments on Lieberman's article: Cyclosiloxanes Produce Fatal Liver and Lung Damage in Mice. Environ Health Perspect 107:161-165 相似文献
10.