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91.
A simplified serum-free method for preparation and cultivation of human granulosa-luteal cells 总被引:1,自引:0,他引:1
Figenschau Y; Sundsfjord JA; Yousef MI; Fuskevag OM; Sveinbjornsson B; Bertheussen K 《Human reproduction (Oxford, England)》1997,12(3):523-531
A simplified method for the preparation and long-term cultivation of
granulosa-luteal cells in serum-free medium is described. The cells were
harvested from women undergoing in-vitro fertilization, enriched by
sedimentation and dissociated by enzymatic treatment. We demonstrated, by
introducing a synthetic serum replacement (SSR2), that these primary cell
cultures cultivated in monolayers on an extracellular matrix may be used in
experiments exceeding 7 days with low cell loss and cell death. No adverse
effect on progesterone production was found. There was a high diversity in
progesterone production between cells from individual patients. After
several days in culture, the cells were challenged with human chorionic
gonadotrophin which revived the rapidly decreasing progesterone production.
We were unable to demonstrate an increase in cell number after 7 days of
cultivation when the cells were grown in medium supplemented with either
serum or SSR2. The mitogens epidermal growth factor and basic fibroblast
growth factor had no influence on proliferation. We also found that the
present method prevents leukocyte contamination in the granulosa-luteal
cell cultures. Compared with the common method based on the enrichment of
granulosa-luteal cells on a density gradient (Ficoll/Percoll), this method
saves time, labour and expense, in addition to augmenting purity.
相似文献
92.
S.K. GUPTA OM SINGH G.P. TALWAR 《American journal of reproductive immunology (New York, N.Y. : 1989)》1985,7(3):104-108
ABSTRACT: Mouse hybrid cell clones secreting antigonadotropin releasing hormone monoclonal antibody were developed by fusion of SP2/O-Ag 1.4 myeloma cells with splenocytes of mouse immunized with gonadotropin releasing hormone (GnRH) tagged to tetanus toxoid. The product of hybrid cell clones obtained as ascites fluid from mouse peritoneal cavity had a titre of 106 (30–40% binding of 125I-GnRH) in radioimmunoassay. The antibody was IgG2a and Kappa. The association constant (Ka) of the product of hybrid cell clone P81662 for binding with GnRH was 1.2 × 109 L/mole. The monoclonal antibody (P81662) was highly specific for the native GnRH and devoid of reactivity with thyroid releasing hormone as tested in competitive radioimmunoassay. The recognition for GnRH agonists by monoclonal was 387-fold less with D-Ser (But)6 des Gly10 GnRH ethylamide and 608-fold less with Bz1-His6 GnRH. Monoclonal anti-GnRH antibody was competent to neutralize the in vivo bioactivity of the hormone as evident by the block of estrus cycle and termination of pregnancy in mice. Termination of pregnancy in animals receiving anti-GnRH monoclonal could be prevented by administration of progesterone. 相似文献
93.
94.
Objective
To evaluate antidiabetic and hypolipidemic activities of Kigelia pinnata methanolic flowers extract in streptozotocin (STZ) induced diabetic wistar rat.Methods
Rats were made diabetic by a single dose of STZ at 60 mg/kg body weight i.p. The blood glucose level was checked before and 72 h after STZ injection to confirm the development of diabetes. The flower extract and glibenclamide were administered orally at the doses of 250 and 500 mg/kg body weight for 21 days.Results
Daily oral treatment with the extract and standard drug for 21 days significantly reduced blood glucose, serum cholesterol and triglycerides levels. High density lipoprotein-cholesterol level was found to be improved (P<0.01) as compared to diabetic control group.Conclusions
It is concluded that Kigellia pinnata flowers extract have significant antidiabetic and hypolipidemic effect. 相似文献95.
Willer CJ Bonnycastle LL Conneely KN Duren WL Jackson AU Scott LJ Narisu N Chines PS Skol A Stringham HM Petrie J Erdos MR Swift AJ Enloe ST Sprau AG Smith E Tong M Doheny KF Pugh EW Watanabe RM Buchanan TA Valle TT Bergman RN Tuomilehto J Mohlke KL Collins FS Boehnke M 《Diabetes》2007,56(1):256-264
More than 120 published reports have described associations between single nucleotide polymorphisms (SNPs) and type 2 diabetes. However, multiple studies of the same variant have often been discordant. From a literature search, we identified previously reported type 2 diabetes-associated SNPs. We initially genotyped 134 SNPs on 786 index case subjects from type 2 diabetes families and 617 control subjects with normal glucose tolerance from Finland and excluded from analysis 20 SNPs in strong linkage disequilibrium (r(2) > 0.8) with another typed SNP. Of the 114 SNPs examined, we followed up the 20 most significant SNPs (P < 0.10) on an additional 384 case subjects and 366 control subjects from a population-based study in Finland. In the combined data, we replicated association (P < 0.05) for 12 SNPs: PPARG Pro12Ala and His447, KCNJ11 Glu23Lys and rs5210, TNF -857, SLC2A2 Ile110Thr, HNF1A/TCF1 rs2701175 and GE117881_360, PCK1 -232, NEUROD1 Thr45Ala, IL6 -598, and ENPP1 Lys121Gln. The replication of 12 SNPs of 114 tested was significantly greater than expected by chance under the null hypothesis of no association (P = 0.012). We observed that SNPs from genes that had three or more previous reports of association were significantly more likely to be replicated in our sample (P = 0.03), although we also replicated 4 of 58 SNPs from genes that had only one previous report of association. 相似文献
96.
97.
Antibody-induced antigenic modulation (AIAM) of CD10 and CD19 was studied on NALM-6, RAJI, and JOK-1 cell lines using fluorescence microscopy (FM), flow cytometry (FCM), and immunoelectron microscopy (IEM). Cross-linking with monoclonal antibodies (MoAbs) induced rapid redistribution of CD10 and CD19 on the cell surface (FM) followed by internalization involving uptake through plasmalemmal pits, transfer through endosomal compartment (receptor-mediated endocytosis), and, finally, delivery to lysosomes for degradation or exocytosis and recycling (IEM). Significant quantitative differences regarding modulation and intracellular processing were shown by FCM and IEM. Thus, 35%, 30%, and 25% of CD10 compared with 80%, 60%, and 40% of CD19 were internalized in NALM-6, RAJI, and JOK-1 cells, respectively. Also, the rate of intracellular transfer as well as externalization and recycling was more pronounced in the case of CD19 than of CD10 and in the NALM-6 and RAJI cells compared with the JOK-1 cells. These differences may possibly reflect the functional significance of CD10 and CD19 as well as the stage of differentiation of the malignant B cells. Although both antigens can be useful in MoAb-targeted immunotherapy, our findings suggest that anti-CD19 MoAbs would be preferable for delivery of cytotoxic agents to malignant B cells. 相似文献
98.
The progesterone receptor gene maps to human chromosome band 11q13, the site of the mammary oncogene int-2. 总被引:3,自引:0,他引:3
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M L Law F T Kao Q Wei J A Hartz G L Greene T Zarucki-Schulz O M Conneely C Jones T T Puck B W O'Malley et al. 《Proceedings of the National Academy of Sciences of the United States of America》1987,84(9):2877-2881
Progesterone is involved in the development and progression of breast cancers, and progesterone receptors (PR) are important markers of hormone dependence and disease prognosis. We have used a human PR cDNA probe, genomic DNA blotting of a series of Chinese hamster-human cell hybrids, and in situ hybridization to map the human PR gene to chromosome 11, band q13. This band also contains the human homolog of the mouse mammary tumor virus integration site, int-2, which surrounds a protooncogene thought to be involved in the development of murine mammary cancers. That these two genes share the same chromosomal location raises important questions about their possible linkage and about the relationship between the mammary-specific oncogene and the steroid hormone in the development, growth, and hormone dependence of human breast cancers. 相似文献
99.
100.
Richard T. Barfield Lynn M. Almli Varun Kilaru Alicia K. Smith Kristina B. Mercer Richard Duncan Torsten Klengel Divya Mehta Elisabeth B. Binder Michael P. Epstein Kerry J. Ressler Karen N. Conneely 《Genetic epidemiology》2014,38(3):231-241
DNA methylation is an important epigenetic mechanism that has been linked to complex diseases and is of great interest to researchers as a potential link between genome, environment, and disease. As the scale of DNA methylation association studies approaches that of genome‐wide association studies, issues such as population stratification will need to be addressed. It is well‐documented that failure to adjust for population stratification can lead to false positives in genetic association studies, but population stratification is often unaccounted for in DNA methylation studies. Here, we propose several approaches to correct for population stratification using principal components (PCs) from different subsets of genome‐wide methylation data. We first illustrate the potential for confounding due to population stratification by demonstrating widespread associations between DNA methylation and race in 388 individuals (365 African American and 23 Caucasian). We subsequently evaluate the performance of our PC‐based approaches and other methods in adjusting for confounding due to population stratification. Our simulations show that (1) all of the methods considered are effective at removing inflation due to population stratification, and (2) maximum power can be obtained with single‐nucleotide polymorphism (SNP)‐based PCs, followed by methylation‐based PCs, which outperform both surrogate variable analysis and genomic control. Among our different approaches to computing methylation‐based PCs, we find that PCs based on CpG sites chosen for their potential to proxy nearby SNPs can provide a powerful and computationally efficient approach to adjust for population stratification in DNA methylation studies when genome‐wide SNP data are unavailable. 相似文献