全文获取类型
收费全文 | 1121篇 |
免费 | 118篇 |
国内免费 | 56篇 |
出版年
2024年 | 5篇 |
2023年 | 17篇 |
2022年 | 34篇 |
2021年 | 50篇 |
2020年 | 42篇 |
2019年 | 51篇 |
2018年 | 66篇 |
2017年 | 34篇 |
2016年 | 60篇 |
2015年 | 71篇 |
2014年 | 74篇 |
2013年 | 80篇 |
2012年 | 95篇 |
2011年 | 95篇 |
2010年 | 57篇 |
2009年 | 57篇 |
2008年 | 51篇 |
2007年 | 49篇 |
2006年 | 46篇 |
2005年 | 23篇 |
2004年 | 22篇 |
2003年 | 28篇 |
2002年 | 26篇 |
2001年 | 26篇 |
2000年 | 12篇 |
1999年 | 9篇 |
1998年 | 11篇 |
1997年 | 7篇 |
1996年 | 5篇 |
1995年 | 5篇 |
1994年 | 7篇 |
1993年 | 3篇 |
1992年 | 8篇 |
1991年 | 13篇 |
1990年 | 4篇 |
1989年 | 7篇 |
1988年 | 8篇 |
1987年 | 8篇 |
1986年 | 2篇 |
1985年 | 3篇 |
1984年 | 4篇 |
1982年 | 1篇 |
1981年 | 2篇 |
1980年 | 4篇 |
1979年 | 4篇 |
1977年 | 1篇 |
1976年 | 4篇 |
1975年 | 1篇 |
1974年 | 3篇 |
排序方式: 共有1295条查询结果,搜索用时 15 毫秒
91.
Jian Shen Baorang Zhang Hua Wei Chuanyan Che Dezhong Ding Xiuguo Hua Peter Bucheli Linghua Wang Yixue Li Xiaoyan Pang Liping Zhao 《Archives of microbiology》2010,192(11):959-968
We first used human flora–associated (HFA) piglets, a significantly improved model for research on human gut microbiota, to study the effects of short-chain fructo-oligosaccharides (scFOS) on the gut bacterial populations. Ten neonatal HFA piglets were assigned to receive basal diets alone or supplemented with scFOS (0.5 g/kg body weight/day) from 1 to 37 days after birth (DAB). The impact of scFOS on the fecal bacterial populations of the piglets before (12 DAB), during (17 DAB), and after (25 and 37 DAB) weaning were monitored by PCR-denaturing gradient gel electrophoresis and real-time quantitative PCR. The Bifidobacterium genus was stimulated consistently, except during weaning, confirming the bifidogenic property of scFOS. At 12 DAB, the Clostridium leptum subgroup was decreased and two unknown Bacteroides-related species were increased; at 25 DAB, the C. leptum subgroup and Subdoligranulum variabile-like species were elevated, whereas one unknown Faecalibacterium-related species was suppressed; and at 37 DAB, the Bacteroides genus was decreased. The results showed that effects of scFOS on non-bifidobacteria varied at different developmental stages of the animals, warranting further investigation into the host-development-related effects of prebiotics on the gut microbiota and the host physiology using the HFA piglets as a model for humans. 相似文献
92.
Yi Liang Jia Cheng Lin Kun Wang Yi Jie Chen Hong Hong Liu Rong Luan Shuai Jiang Tao Che Yong Zhao De Feng Li Da Cheng Wang Lin Guo Hui Sun 《Biochimica et Biophysica Acta (BBA)/General Subjects》2010
Background
We have previously reported a novel fungal galectin Agrocybe aegerita lectin (AAL) with apoptosis-induced activity and nuclear migration activity. The importance of nuclear localization for AAL's apoptosis-induced activity has been established by mutant study. However, the mechanism remains unclear.Methods
We further investigated the mechanism using a previously reported carbohydrate recognition domain (CRD) mutant protein H59Q, which retained its nuclear localization activity but lost most of its apoptotic activity. The cell membrane-binding ability of recombinant AAL (rAAL) and H59Q was analyzed by FACS, and their cellular partners were identified by affinity chromatography and mass spectroscopy. Furthermore, the interaction of AAL and ligand was proved by mammalian two-hybrid and pull down assays. A knockdown assay was used to confirm the role of the ligand.Results
The apoptotic activity of AAL could be blocked by lactose. Mutant H59Q retained comparable cell membrane-binding ability to rAAL. Four cellular binding partners of AAL in HeLa cells were identified: glucose-regulated protein 78 (GRP78); mortality factor 4-like protein 1 (MRG15); elongation factor 2 (EEF2); and heat shock protein 70 (Hsp70). CRD region of AAL was required for the interaction between AAL/mutant AAL and MRG15. MRG15 knockdown increased the cells' resistance to AAL treatment.Conclusion
MRG15 was a nuclear ligand for AAL in HeLa cells. These data implied the existence of a novel nuclear pathway for the antitumor activity of fungal galectin AAL.General significance
These findings provide a novel explanation of AAL bioactivity and contribute to the understanding of mushroom lectins' antitumor activity. 相似文献93.
Shu‐Jyuan Yang Szu‐Min Chang Kun‐Che Tsai Wen‐Shiang Chen Feng‐Huei Lin Ming‐Jium Shieh 《The journal of gene medicine》2010,12(2):168-179
Background
Gene therapy has been used to treat a variety of health problems, but transfection inefficiency and the lack of safe vectors have limited clinical progress. Fabrication of a vector that is safe and has high transfection efficiency is crucial for the development of successful gene therapy. The present study aimed to synthesize chitosan‐alginate nanoparticles that can be used as carriers of the pAcGFP1‐C1 plasmid and to use these nanoparticles with an ultrasound protocol to achieve high efficiency gene transfection.Methods
Chitosan was complexed with alginate and the pAcGFP1‐C1 plasmid at different charge ratios to create chitosan‐alginate‐DNA nanoparticles (CADNs). The average particle size and loading efficiency were measured. Plasmid DNA retardation and integrity were analysed on 1% agarose gels. The effect of CADNs and ultrasound on the efficiency of transfection of cells and subcutaneous tumors was evaluated.Results
In the CADNs, the average size of incorporated plasmid DNA was 600–650 nm and the loading efficiency was greater than 90%. On the basis of the results of the plasmid DNA protection test, CADNs could protect the transgene from DNase I degradation. The transgene product expression could be enhanced efficiently if cells or tumor tissues were first given CADNs and then treated with ultrasound.Conclusions
The use of CADNs combined with an ultrasound regimen is a promising method for safe and effective gene therapy. Copyright © 2009 John Wiley & Sons, Ltd.94.
The development of ACF (aberrant crypt foci), adenoma and cancer following intrarectal administration of the alkylating agent N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) has been described. However, microscopic lesions not previously reported were observed as soon as two weeks following carcinogen treatment. These lesions protrude slightly over the epithelial lining of the colon, with a micropolyp-like appearance. Oriented sections show that the centre of these lesions present pseudo-“cystic” appearance, with disorganized crypts made of normal cells. The chorion of the lesion is invaded by numerous inflammatory cells and some ACF may be present nearby. The epithelium lining the cysts and the distorted crypts shows expression of gastric mucin M1/MUC5AC, an early marker of colonic carcinogenesis which is not present in normal colon. This mucin is retained within the “cysts” together with some inflammatory cells.The micropolyps observed contain in a minute form some histological elements described in ulcerative colitis or short-term radiotherapy (distortion of crypts, crypt abscesses, increase of chorion cellularity, infiltration by immune cells). In addition, the presence of bifid crypts nearby suggests mucosal regeneration.Our hypothesis is that these modifications are steps in a normal healing pathway that may in some cases degenerate into precancerous lesions and cancer. 相似文献
95.
96.
Chemical activation of parthenogenetic and nuclear transfer porcine oocytes using ionomycin and strontium chloride 总被引:5,自引:0,他引:5
Effective protocols for oocyte activation are crucial for study of parthenogenetic development and to produce nuclear transfer reconstructed embryos. This study investigated the use of ionomycin (ION) and strontium chloride (Sr(2+)) in the activation of parthenogenetic and nuclear transfer porcine oocytes. In-vitro-matured oocytes with a polar body were treated with varying concentrations of ION, Sr(2+) or its combinations, and then fixed or cultured to assess activation and development rates, respectively. Ionomycin concentrations of 10 and 15 microM resulted in more frequent oocyte activation and the 15 microM in advanced development compared to 5 microM (71.8 and 70%vs. 47.5%; P=0.04, and 43.7%vs. 19.3%; P=0.008, respectively). Oocytes treated with 10, 20 or 30 mM of Sr(2+) for 2 or 4h displayed a pronuclear formation rate ranging from 46.7 to 70%. When employed after a 5 min treatment with 10 or 15 microM ION, exposure to 10 mM Sr(2+) for 4 h resulted in higher pronuclear formation than did the 20 mM concentration (82 and 88.6%vs. 63.3 and 73.2%; P=0.03). Nuclear transfer reconstructed oocytes treated with 15 microM/5 min ION followed by 10 mM/4 h Sr(2+) resulted in a higher development to blastocyst stage compared to those treated with 15 microM ION alone (17.7 vs. 11.3%; P=0.06). In conclusion, we inferred that the inclusion of Sr(2+) in the activation protocol can benefit the development of nuclear transfer reconstructed porcine oocytes. 相似文献
97.
Although the results of some studies indicate that salt stress affects the organization of microtubules, it remains an open question whether microtubules play an active role in the plant's ability to withstand salt stress. In the present study, we showed that salt stress-induced wild-type Arabidopsis seedling roots display right-handed skewed growth and depolymerization of the cortical microtubules. The results of a long-term observational study showed that cortical microtubules depolymerized then reorganized themselves under salt stress. Stabilization of microtubules with paclitaxel resulted in more seedling death under salt stress, while disruption of microtubules with oryzalin or propyzamide rescued seedlings from death. Seedlings in which the cortical microtubules were reorganized did not succumb to salt stress. These results suggest that both depolymerization and reorganization of the cortical microtubules are important for the plant's ability to withstand salt stress. Depolymerizing microtubules by drugs rescues seedlings from death under salt stress. This rescue effect was abolished by removing calcium from the medium or treatment with a calcium channel inhibitor. Depolymerization of the microtubules is followed by an increase in the free cytoplasmic calcium concentration. The addition of calcium to the growth medium increased the number of seedlings in which recovery of the cortical microtubules occurred, whereas the removal of calcium decreased the number of seedlings in which recovery occurred. Therefore, depolymerization of the cortical microtubules raises intracellular calcium concentrations, while reorganization of the cortical microtubules and seedling survival may be mediated by calcium influx in salt stress. 相似文献
98.
Arabidopsis shoots regenerate from root explants in tissue culture through a two-step process requiring preincubation on an auxin-rich
callus induction medium (CIM) followed by incubation on a cytokinin-rich shoot induction medium (SIM). During CIM preincubation,
root explants acquire competence to respond to shoot induction signals. During CIM preincubation, pericycle cells in root
explants undergo cell divisions and dedifferentiate, losing the expression of a pericycle cell-specific marker. These cells
acquire competence to form green callus only after one day CIM preincubation and to form shoots after 2–3 days CIM preincubation.
Reversible DNA synthesis inhibitors interfered with the acquisition of competence to form shoots. Genes requiring CIM preincubation
for upregulation on SIM were identified by microarray analysis and included RESPONSE REGULATOR 15 (ARR15), POLYGALACTURONASE INHIBITING PROTEIN 2 (PGIP2) and WUSCHEL (WUS). These genes served as developmental markers for the acquisition of competence because the CIM preincubation requirements
for ARR15 and PGIP2 upregulation correlated well with the acquisition of competence to form green callus, and the CIM preincubation requirements
for WUS upregulation matched those for shoot formation. Unlike ARR15, another cytokinin inducible, A-type ARR gene, ARR5, was upregulated on SIM, but the induction did not require CIM preincubation. These findings indicate that competencies for
various events associated with shoot regeneration are acquired progressively during CIM preincubation, and that a set of genes,
normally upregulated on SIM, are repressed by a process that can be relieved by CIM preincubation. 相似文献
99.
100.
1-Hydroxy-2, 3, 5-trimethoxyxanthone (HM-1) is a xanthone isolated from Halenia elliptica, a Tibetan medicinal herb. HM-1 (0.33-42.1 microM) produced a concentration-dependent relaxation in rat coronary artery rings pre-contracted with 1 microM 5-hydroxytryptamine (5-HT), with an EC(50) of 1.67+/-0.27 microM. Removal of the endothelium significantly affected the vasodilator potency of HM-1, resulting in 46% decrease in E(max) value. The endothelium-dependent effects of HM-1 was confirmed when its vasorelaxant effect was inhibited after addition of nitric oxide synthase (NOS) inhibitor N(omega)-nitro-l-arginine methyl ester (100 microM) or the soluble guanylate cyclase inhibitor 1H-[1, 2, 4] oxadiazolo [4,3-alpha] quinoxalin-1-one (ODQ, 10 microM). Atropine (100 nM), flurbiprofen (10 microM), propranolol (100 microM), pyrilamine (10 microM), cimetidine (10 microM) and SQ22536 (100 microM) had no effect on the vasorelaxant activity of HM-1 indicated the non-involvement of other receptor/enzyme systems. In endothelium-denuded coronary artery rings, the vasorelaxant effect of HM-1 was unaffected by potassium channel blockers such as tetraethylammonium (10 mM), iberiotoxin (100 nM), barium chloride (100 microM) and 4-aminopyridine (1 mM). The involvement of Ca(2+) channel in 5-HT-primed artery ring preparations incubated with Ca(2+)-free buffer was confirmed when HM-1 (9.93 microM) partially abolished the CaCl(2)-induced vasoconstriction (87% inhibition in intact-endothelium artery rings; 50% inhibition in endothelium-denuded rings). In the KCl-primed preparations incubated with Ca(2+)-free buffer, HM-1 (9.93 microM) produced a 27.3% inhibition in endothelium-denuded rings. HM-1 (3.31-33.1 microM) had minimal relaxant effects (14.4%-20.3%) on the contractile response generated by 10 microM phorbol 12,13-diacetate (PDA) in Ca(2+)-free solutions, suggesting minimal effects on intracellular Ca(2+) mechanisms. These findings suggest the vasodilator action of HM-1 involved both an endothelium-dependent mechanism involving NO and an endothelium-independent mechanism by inhibiting Ca(2+) influx through L-type voltage-operated Ca(2+) channels; a minor contribution to the effects of HM-1 may be related to inhibition of the protein kinase C-mediated release of intracellular Ca(2+) stores. 相似文献