全文获取类型
收费全文 | 1112篇 |
免费 | 46篇 |
专业分类
1158篇 |
出版年
2022年 | 10篇 |
2021年 | 8篇 |
2019年 | 7篇 |
2018年 | 13篇 |
2017年 | 12篇 |
2016年 | 16篇 |
2015年 | 19篇 |
2014年 | 26篇 |
2013年 | 72篇 |
2012年 | 43篇 |
2011年 | 43篇 |
2010年 | 35篇 |
2009年 | 30篇 |
2008年 | 87篇 |
2007年 | 56篇 |
2006年 | 62篇 |
2005年 | 76篇 |
2004年 | 55篇 |
2003年 | 50篇 |
2002年 | 56篇 |
2001年 | 29篇 |
2000年 | 44篇 |
1999年 | 41篇 |
1998年 | 6篇 |
1997年 | 10篇 |
1996年 | 8篇 |
1995年 | 11篇 |
1994年 | 14篇 |
1993年 | 10篇 |
1992年 | 24篇 |
1991年 | 23篇 |
1990年 | 18篇 |
1989年 | 18篇 |
1988年 | 13篇 |
1987年 | 10篇 |
1986年 | 13篇 |
1985年 | 8篇 |
1984年 | 9篇 |
1983年 | 8篇 |
1982年 | 7篇 |
1981年 | 11篇 |
1980年 | 3篇 |
1978年 | 4篇 |
1976年 | 3篇 |
1975年 | 4篇 |
1974年 | 3篇 |
1973年 | 5篇 |
1972年 | 5篇 |
1969年 | 4篇 |
1967年 | 3篇 |
排序方式: 共有1158条查询结果,搜索用时 15 毫秒
171.
Reduction of chlortetracycline-resistant Escherichia coli in weaned piglets fed fermented liquid feed 总被引:2,自引:0,他引:2
We investigated the change in chlortetracycline resistance in 360 Escherichia coli strains separated from the feces of piglets fed fermented liquid feed (FLF) in comparison with those fed dry feed (control). The total amount of lactic acid bacteria in feces was 8.77x10(8)CFU/g DM at weaning, which increased to 1.23x10(12)CFU/g DM (FLF) at 28 days after weaning (P<0.001). As a result of the antibiotic susceptibility test, almost all isolates were chlortetracycline-resistant (CTC(R)) until 14 days after weaning both in the FLF and control groups. At 28 days, the CTC(R)E. coli decreased to 22.2% in the FLF group, while the proportion of resistant bacteria was 88.9% in the control group. In addition, as a result of the gene analysis, it was clarified that there is a relation between the decrease in the minimum inhibitory concentration values and the possession rate of the tetracycline-resistance gene tet(A), tet(B) and tet(D). These results show that FLF caused an increase in the number of lactic acid bacteria in the intestines, and suggested that the feeding of FLF can possibly reduce antibiotic-resistance bacteria. 相似文献
172.
Aim: Ultraviolet (UV) mutagenesis was carried out to obtain mutant strains of Cupriavidus necator that could produce ( R )-3-hydroxybutyric acid [( R )-3-HB] in the culture supernatant.
Methods and Results: C. necator (formerly known as Ralstonia eutropha ) was subjected to UV radiation to generate mutants that are capable of producing ( R )-3-HB in the culture supernatant. Results indicated that UV mutagen disrupted the phbB ( phbB knock-out) and thus, promoted production of ( R )-3-HB in mutant strains. Inclusion of acetoacetate esters (carbonyl compounds) in the culture broth led to increased production of ( R )-3-HB. Thus, acetoacetyl-CoA (an intermediate of the PHB synthetic pathway) might have been converted to acetoacetate, which in the presence of ( R )-3-HB dehydrogenase and NADPH/NADP+ , resulted in extracellular production of ( R )-3-HB.
Conclusions: UV mutagenesis proved to be a satisfactory method in generating interesting mutants for extracellular production of ( R )-3-HB. Extracellular production of ( R )-3-HB upon addition of acetoacetate esters would suggest a likely ( R )-3-HB biosynthetic pathway in C. necator .
Significance and Impact of the Study: Mutants obtained in this study are very useful for production of ( R )-3-HB. For the first time, the production of ( R )-3-HB by C. necator via acetoacetate is reported. 相似文献
Methods and Results: C. necator (formerly known as Ralstonia eutropha ) was subjected to UV radiation to generate mutants that are capable of producing ( R )-3-HB in the culture supernatant. Results indicated that UV mutagen disrupted the phbB ( phbB knock-out) and thus, promoted production of ( R )-3-HB in mutant strains. Inclusion of acetoacetate esters (carbonyl compounds) in the culture broth led to increased production of ( R )-3-HB. Thus, acetoacetyl-CoA (an intermediate of the PHB synthetic pathway) might have been converted to acetoacetate, which in the presence of ( R )-3-HB dehydrogenase and NADPH/NADP
Conclusions: UV mutagenesis proved to be a satisfactory method in generating interesting mutants for extracellular production of ( R )-3-HB. Extracellular production of ( R )-3-HB upon addition of acetoacetate esters would suggest a likely ( R )-3-HB biosynthetic pathway in C. necator .
Significance and Impact of the Study: Mutants obtained in this study are very useful for production of ( R )-3-HB. For the first time, the production of ( R )-3-HB by C. necator via acetoacetate is reported. 相似文献
173.
Fujibayashi A Taguchi T Misaki R Ohtani M Dohmae N Takio K Yamada M Gu J Yamakami M Fukuda M Waguri S Uchiyama Y Yoshimori T Sekiguchi K 《Cell structure and function》2008,33(1):35-50
RME-8 is a DnaJ-domain-containing protein that was first identified in Caenorhabditis elegans as being required for uptake of yolk proteins. RME-8 has also been identified in other species, including flies and mammals, and the phenotypes of their RME-8 mutants suggest the importance of this protein in endocytosis. In the present study, we cloned human RME-8 (hRME-8) and characterized its biochemical properties and functions in endocytic pathways. hRME-8 was found to be a peripheral protein that was tightly associated with the membrane via its N-terminal region. It partially colocalized with several early endosomal markers, but not with late endosomal markers, consistent with observations by immunoelectron microscopy. When cells were transfected with a panel of dominant-active Rab proteins, hRME-8 was confined to large vacuoles induced by expression of Rab5aQ79L, but not by Rab7Q67L. Expression of C-terminally-truncated hRME-8 mutants led to the formation of large puncta and vacuoles, and compromised endocytic pathways through early endosomes, i.e., recycling of transferrin and degradation of epidermal growth factor. Taken together, these results indicate that hRME is primarily involved in membrane trafficking through early endosomes, but not through degradative organelles, such as multivesicular bodies and late endosomes. 相似文献
174.
Tashiro Y Nomura N Nakao R Senpuku H Kariyama R Kumon H Kosono S Watanabe H Nakajima T Uchiyama H 《Journal of bacteriology》2008,190(11):3969-3978
Pseudomonas aeruginosa is an opportunistic bacterial pathogen that is one of the most refractory to therapy when it forms biofilms in the airways of cystic fibrosis patients. To date, studies regarding the production of an immunogenic and protective antigen to inhibit biofilm formation by P. aeruginosa have been superficial. The previously uncharacterized outer membrane protein (OMP) Opr86 (PA3648) of P. aeruginosa is a member of the Omp85 family, of which homologs have been found in all gram-negative bacteria. Here we verify the availability of Opr86 as a protective antigen to inhibit biofilm formation by P. aeruginosa PAO1 and several other isolates. A mutant was constructed in which Opr86 expression could be switched on or off through a tac promoter-controlled opr86 gene. The result, consistent with previous Omp85 studies, showed that Opr86 is essential for viability and plays a role in OMP assembly. Depletion of Opr86 resulted in streptococci-like morphological changes and liberation of excess membrane vesicles. A polyclonal antibody against Opr86 which showed reactivity to PAO1 cells was obtained. The antibody inhibited biofilm formation by PAO1 and the other clinical strains tested. Closer examination of early attachment revealed that cells treated with the antibody were unable to attach to the surface. Our data suggest that Opr86 is a critical OMP and a potential candidate as a protective antigen against biofilm formation by P. aeruginosa. 相似文献
175.
Chikatoshi Yanagimoto Masaru Harada Hiroto Kumemura Takumi Kawaguchi Shinichiro Hanada Yukio Koizumi Haruaki Ninomiya Toshihiro Sugiyama 《Experimental cell research》2009,315(2):119-126
Wilson disease is a genetic disorder characterized by the accumulation of copper in the body by defective biliary copper excretion. Wilson disease gene product (ATP7B) functions in copper incorporation to ceruloplasmin (Cp) and biliary copper excretion. However, copper metabolism in hepatocytes has been still unclear. Niemann-Pick disease type C (NPC) is a lipid storage disorder and the most commonly mutated gene is NPC1 and its gene product NPC1 is a late endosome protein and regulates intracellular vesicle traffic. In the present study, we induced NPC phenotype and examined the localization of ATP7B and secretion of holo-Cp, a copper-binding mature form of Cp. The vesicle traffic was modulated using U18666A, which induces NPC phenotype, and knock down of NPC1 by RNA interference. ATP7B colocalized with the late endosome markers, but not with the trans-Golgi network markers. U18666A and NPC1 knock down decreased holo-Cp secretion to culture medium, but did not affect the secretion of other secretory proteins. Copper accumulated in the cells after the treatment with U18666A. These findings suggest that ATP7B localizes in the late endosomes and that copper in the late endosomes is transported to the secretory compartment via NPC1-dependent pathway and incorporated into apo-Cp to form holo-Cp. 相似文献
176.
Four types of DNA polymerase (Pol beta, Pol lambda, Pol mu and TdT) have been identified in eukaryotes as members of the polymerase X-family. Only vertebrates have all four types of enzyme. Plants and fungi have one or two X-family polymerases, while protostomes, such as fruit flies and nematodes, do not appear to have any. It is possible that the well-known metabolic pathways in which these enzymes are involved are restricted to the vertebrate world. The distribution of the DNA polymerases involved in DNA repair across the various biological kingdoms differs from that of the DNA polymerases involved in chromosomal DNA replication. In this review, we focus on the interesting pattern of distribution of the X-family enzymes across biological kingdoms and speculate on their roles. 相似文献
177.
Tetsuro Tamaki Yoshiyasu Uchiyama Yoshinori Okada Kayoko Tono Masahiro Nitta Akio Hoshi Akira Akatsuka 《Histochemistry and cell biology》2009,132(1):59-70
Tissue inflammation and multiple cellular responses in the compensatory enlarged plantaris (OP Plt) muscle induced by surgical
ablation of synergistic muscles (soleus and gastrocnemius) were followed over 10 weeks after surgery. Contralateral surgery
was performed in adult Wistar male rats. Cellular responses in muscle fibers, blood vessels and nerve fibers were analyzed
by immunohistochemistry and electron microscopy. Severe muscle fiber damage and disappearance of capillaries associated with
apparent tissue edema were observed in the peripheral portion of OP Plt muscles during the first week, whereas central portions
were relatively preserved. Marked cell activation/proliferation was also mainly observed in peripheral portions. Similarly,
activated myogenic cells were seen not only inside but also outside of muscle fibers. The former were likely satellite cells
and the latter may be interstitial myogenic cells. One week after surgery, small muscle fibers, small arteries and capillaries
and several branched-muscle fibers were evident in the periphery, thus indicating new muscle fiber and blood vessel formation.
Proliferating cells were also detected in the nerve bundles in the Schwann cell position. These results indicate that the
compensatory stimulated/enlarged muscle is a suitable model for analyzing multiple physiological cellular responses in muscle–nerve–blood
vessel units under continuous stretch stimulation.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
178.
Kensuke Kobayashi Minaho Uchiyama Hirobumi Takahashi Hiroshi Kawamoto Satoru Ito Takashi Yoshizumi Hiroshi Nakashima Tetsuya Kato Atsushi Shimizu Izumi Yamamoto Masanori Asai Hiroshi Miyazoe Akio Ohno Mioko Hirayama Satoshi Ozaki Takeshi Tani Yasuyuki Ishii Takeshi Tanaka Takanobu Mochidome Kiyoshi Tadano Osamu Okamoto 《Bioorganic & medicinal chemistry letters》2009,19(11):3096-3099
The synthesis and biological evaluation of new potent opioid receptor-like 1 (ORL1) antagonists are presented. Conversion of the thioether linkage of the prototype [It is reported prior to this communication as a consecutive series.: Kobayashi, K.; Kato, T.; Yamamoto, I.; Shimizu, A.; Mizutani, S.; Asai, M.; Kawamoto, H.; Ito, S.; Yoshizumi, T.; Hirayama, M.; Ozaki, S.; Ohta, H.; Okamoto, O. Bioorg. Med. Chem. Lett., in press] to the carbonyl linker effectively reduces susceptibility to P-glycoprotein (P-gp) efflux. This finding led to the identification of 2-cyclohexylcarbonylbenzimizole analogue 7c, which exhibited potent ORL1 activity, excellent selectivity over other receptors and ion channels, and poor susceptibility to P-gp. Compound 7c also showed satisfactory pharmacokinetic profiles and brain penetrability in laboratory animals. Furthermore, 7c showed good in vivo antagonism. Hence, 7c was selected as a clinical candidate for a brain-penetrable ORL1 antagonist. 相似文献
179.
180.
Ken Urabe Kouji Naruse Masataka Uchino Masashi Takaso Mamoru Fujita Katsufumi Uchiyama Takamitsu Okada Midori Kasahara Moritoshi Itoman 《Cell and tissue banking》2009,10(3):259-265
Demand for banked bone allografts is increasing in Japan; however, there are too few bone banks and the bone bank network
is not well-established. One reason for this was lack of funding for banks. Bone banks had to bear all material expenses of
banked bone allografts themselves because this was not designated a covered expense. In December 2004, the Japanese government
started a new “Advanced Medical Treatment” administration system which allowed an approved institution to charge the expense
of authorized advanced medical treatments directly to patients. The treatment named “Cryopreserved allogenic bone and ligamentous
tissue retrieved from cadaveric donor” was approved as an advanced medical treatment in March 2007. We present the calculation
method and the expense per implantation of a banked bone allograft from a cadaveric donor under this treatment and raise issues
which affect this advanced medical treatment and remain to be resolved in the Japanese orthopaedic field. 相似文献