全文获取类型
收费全文 | 850篇 |
免费 | 74篇 |
出版年
2021年 | 4篇 |
2020年 | 4篇 |
2019年 | 6篇 |
2018年 | 6篇 |
2017年 | 15篇 |
2016年 | 19篇 |
2015年 | 32篇 |
2014年 | 31篇 |
2013年 | 48篇 |
2012年 | 55篇 |
2011年 | 64篇 |
2010年 | 38篇 |
2009年 | 27篇 |
2008年 | 48篇 |
2007年 | 57篇 |
2006年 | 35篇 |
2005年 | 45篇 |
2004年 | 50篇 |
2003年 | 42篇 |
2002年 | 42篇 |
2001年 | 15篇 |
2000年 | 13篇 |
1999年 | 16篇 |
1998年 | 14篇 |
1997年 | 11篇 |
1996年 | 7篇 |
1995年 | 7篇 |
1994年 | 9篇 |
1993年 | 10篇 |
1992年 | 11篇 |
1991年 | 9篇 |
1990年 | 12篇 |
1989年 | 4篇 |
1988年 | 6篇 |
1987年 | 11篇 |
1986年 | 6篇 |
1985年 | 7篇 |
1984年 | 7篇 |
1983年 | 9篇 |
1982年 | 4篇 |
1981年 | 8篇 |
1980年 | 8篇 |
1979年 | 6篇 |
1978年 | 7篇 |
1977年 | 5篇 |
1973年 | 4篇 |
1972年 | 4篇 |
1971年 | 4篇 |
1969年 | 3篇 |
1954年 | 2篇 |
排序方式: 共有924条查询结果,搜索用时 15 毫秒
1.
Guy Froyen An Broekmans Femke Hillen Karin Pat Ruth Achten Jeroen Mebis Jean-Luc Rummens Johan Willemse Brigitte Maes 《PloS one》2016,11(4)
The inevitable switch from standard molecular methods to next-generation sequencing for the molecular profiling of tumors is challenging for most diagnostic laboratories. However, fixed validation criteria for diagnostic accreditation are not in place because of the great variability in methods and aims. Here, we describe the validation of a custom panel of hotspots in 24 genes for the detection of somatic mutations in non-small cell lung carcinoma, colorectal carcinoma and malignant melanoma starting from FFPE sections, using 14, 36 and 5 cases, respectively. The targeted hotspots were selected for their present or future clinical relevance in solid tumor types. The target regions were enriched with the TruSeq approach starting from limited amounts of DNA. Cost effective sequencing of 12 pooled libraries was done using a micro flow cell on the MiSeq and subsequent data analysis with MiSeqReporter and VariantStudio. The entire workflow was diagnostically validated showing a robust performance with maximal sensitivity and specificity using as thresholds a variant allele frequency >5% and a minimal amplicon coverage of 300. We implemented this method through the analysis of 150 routine diagnostic samples and identified clinically relevant mutations in 16 genes including KRAS (32%), TP53 (32%), BRAF (12%), APC (11%), EGFR (8%) and NRAS (5%). Importantly, the highest success rate was obtained when using also the low quality DNA samples. In conclusion, we provide a workflow for the validation of targeted NGS by a custom-designed pan-solid tumor panel in a molecular diagnostic lab and demonstrate its robustness in a clinical setting. 相似文献
2.
3.
Andrew J. Wright Marie Maar Christian Mohn Jacob Nabe-Nielsen Ursula Siebert Lasse Fast Jensen Hans J. Baag?e Jonas Teilmann 《PloS one》2013,8(2)
An unprecedented 85 harbour porpoises stranded freshly dead along approximately 100 km of Danish coastline from 7–15 April, 2005. This total is considerably above the mean weekly stranding rate for the whole of Denmark, both for any time of year, 1.23 animals/week (ranging from 0 to 20 during 2003–2008, excluding April 2005), and specifically in April, 0.65 animals/week (0 to 4, same period). Bycatch was established as the cause of death for most of the individuals through typical indications of fisheries interactions, including net markings in the skin and around the flippers, and loss of tail flukes. Local fishermen confirmed unusually large porpoise bycatch in nets set for lumpfish (Cyclopterus lumpus) and the strandings were attributed to an early lumpfish season. However, lumpfish catches for 2005 were not unusual in terms of season onset, peak or total catch, when compared to 2003–2008. Consequently, human activity was combined with environmental factors and the variation in Danish fisheries landings (determined through a principal component analysis) in a two-part statistical model to assess the correlation of these factors with both the presence of fresh strandings and the numbers of strandings on the Danish west coast. The final statistical model (which was forward selected using Akaike information criterion; AIC) indicated that naval presence is correlated with higher rates of porpoise strandings, particularly in combination with certain fisheries, although it is not correlated with the actual presence of strandings. Military vessels from various countries were confirmed in the area from the 7th April, en route to the largest naval exercise in Danish waters to date (Loyal Mariner 2005, 11–28 April). Although sonar usage cannot be confirmed, it is likely that ships were testing various equipment prior to the main exercise. Thus naval activity cannot be ruled out as a possible contributing factor. 相似文献
4.
Pei W. Thomas Timothy Spicer Michael Cammarata Jennifer S. Brodbelt Peter Hodder Walter Fast 《Bioorganic & medicinal chemistry》2013,21(11):3138-3146
Due to the global threat of antibiotic resistance mediated by New Delhi metallo-beta-lactamase-1 (NDM-1) and the lack of structurally diverse inhibitors reported for this enzyme, we developed screening and counter-screening assays for manual and automated formats. The manual assay is a trans-well absorbance-based endpoint assay in 96-well plates and has a Z′ factor of 0.8. The automated assay is an epi-absorbance endpoint assay in 384-well plates, has a Z′ factor of ?0.8, good signal/baseline ratios (>3.8), and is likely scalable for high-throughput screening (HTS). A TEM-1-based counter-screen is also presented to eliminate false positives due to assay interference or off-target activities. A pilot screen of a pharmacologically characterized compound library identified two thiol-modifying compounds as authentic NDM-1 inhibitors: p-chloromecuribenzoate (p-CMB) and nitroprusside. Recombinant NDM-1 has one Cys residue that serves as a conserved active-site primary zinc ligand and is selectively modified by p-CMB as confirmed by LC–MS/MS. However a C208D mutation results in an enzyme that maintains almost full lactamase activity, yet is completely resistant to the inhibitor. These results predict that covalent targeting of the conserved active-site Cys residue may have drawbacks as a drug design strategy. 相似文献
5.
Occurrence of two serologically distinct groups within Bacillus thuringiensis serotype 3 ab var. kurstaki 总被引:7,自引:0,他引:7
Two groups distinguishable on the basis of crystal serology have been identified within Bacillus thuringiensis var kurstaki (serotype 3 ab). The toxicities of these two groups to Trichoplusia ni and Heliothis virescens expressed as ratio also differed. It is suggested that serotype 3 ab be separated into two subgroups designated as K-1 and K-73. 相似文献
6.
Pat. Howard 《e》2009,4(4):e170-e171
7.
8.
Komal Preet Allagh B. R. Shamanna Gudlavalleti V. S. Murthy Andy R. Ness Pat Doyle Sutapa B. Neogi Hira B. Pant Wellcome Trust- PHFI Folic Acid project team 《PloS one》2015,10(3)
Background
In the last two decades, India has witnessed a substantial decrease in infant mortality attributed to infectious disease and malnutrition. However, the mortality attributed to birth defects remains constant. Studies on the prevalence of birth defects such as neural tube defects and orofacial clefts in India have reported inconsistent results. Therefore, we conducted a systematic review of observational studies to document the birth prevalence of neural tube defects and orofacial clefts.Methods
A comprehensive literature search for observational studies was conducted in MEDLINE and EMBASE databases using key MeSH terms (neural tube defects OR cleft lip OR cleft palate AND Prevalence AND India). Two reviewers independently reviewed the retrieved studies, and studies satisfying the eligibility were included. The quality of included studies was assessed using selected criteria from STROBE statement.Results
The overall pooled birth prevalence (random effect) of neural tube defects in India is 4.5 per 1000 total births (95% CI 4.2 to 4.9). The overall pooled birth prevalence (random effect) of orofacial clefts is 1.3 per 1000 total births (95% CI 1.1 to 1.5). Subgroup analyses were performed by region, time period, consanguinity, and gender of newborn.Conclusion
The overall prevalence of neural tube defects from India is high compared to other regions of the world, while that of orofacial clefts is similar to other countries. The majority of studies included in the review were hospital based. The quality of these studies ranged from low to moderate. Further well-designed, high quality community-based observational studies are needed to accurately estimate the burden of neural tube defects and orofacial clefts in India. 相似文献9.
Etienne Karita Matt A Price Shabir Lakhi William Kilembe Anatoli Kamali Eugene Ruzagira Eric Hunter Paul Farmer Susan Allen Gwynn Stevens Paramesh Chetty Sabrina Welsh Annie Yang Jill Gilmour Pat Fast The IAVI Africa HIV Prevention Partnership 《PloS one》2015,10(8)
Background
2013 WHO guidelines recommend starting ART at CD4+ T-cell counts ≤500 cells/μL. We present the T-cell counts from adult Africans with HIV shortly following transmission to their sexual partners.Methods
HIV-discordant couples in Zambia, Uganda and Rwanda were followed prospectively and received couples counseling and condoms. HIV uninfected partners were tested for HIV at least quarterly and HIV-infected partners received HIV care and referral for ART per national guidelines. Upon diagnosis of incident HIV infection in the previously HIV-uninfected partner, a blood sample was collected from both partners to measure CD4+ T-cells and perform viral linkage. The estimated date of infection (EDI) of the incident case was calculated based on testing history. EDI was unknown for suspected transmitting partners.Results
From 2006–2011, 4,705 HIV-discordant couples were enrolled in this cohort, and 443 cases of incident HIV infection were documented. Virus linkage analysis was performed in 374 transmission pairs, and 273 (73%) transmissions were linked genetically. CD4 counts in the transmitting partner were measured a median of 56 days after EDI (mean:90.5, min:10, max:396). The median CD4 count was 339 cells/μl (mean:386.4, min:15, max:1,434), and the proportion of partners with a CD4+ T-cell count above 500/μl was 25% (95% CI:21, 31).Conclusions
In our cohort of discordant couples, 73% of HIV transmissions occurred within the relationship, and the transmitter CD4+ T cell count shortly after the transmission event was frequently higher than the WHO 2013 ART-initiation guidelines. 相似文献10.
Gloria Omosa-Manyonyi Juliet Mpendo Eugene Ruzagira William Kilembe Elwyn Chomba Fran?ois Roman Patricia Bourguignon Marguerite Koutsoukos Alix Collard Gerald Voss Dagna Laufer Gwynn Stevens Peter Hayes Lorna Clark Emmanuel Cormier Len Dally Burc Barin Jim Ackland Kristen Syvertsen Devika Zachariah Kamaal Anas Eddy Sayeed Angela Lombardo Jill Gilmour Josephine Cox Patricia Fast Frances Priddy 《PloS one》2015,10(5)