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71.
Vertical distribution in and isolation of bacteria from Lake Vanda: an Antarctic lake 总被引:1,自引:1,他引:0
Susumu Takii Toshifumi Konda Akira Hiraishi Genki I. Matsumoto Tamio Kawano Tetsuya Torii 《Hydrobiologia》1986,135(1-2):15-21
Vertical distribution of bacteria in Lake Vanda, an Antarctic meromictic lake, was examined by the acridine orange epifluorescence direct count method. Total bacteria were 104–105 cells · ml–1 in the water at 55 m depth and above, and increased drastically to 107 cells · ml–1 in the bottom water. Filamentous or long rodshaped bacteria occurred at a high frequency in the upper layers, but in the bottom layers most bacteria were coccoidal or short rods. Mean bacterial cell volume in water of between 10 m and 60 m deep was fairly large compared with common bacterial populations in seawater and lake water. Aerobic heterotrophic bacteria were recovered from the water of a depth of 30 m and above, and were assumed to belong to Caulobacter. Viable heterotrophic bacteria were not recovered from the high salinity deep water by media prepared with the same deep water. Phototrophic purple non-sulphur bacteria were isolated by enrichment cultures from water at 55 m depth. 相似文献
72.
Mamoru Harada Koji Tamada Koichiro Abe Tieli Li Yasuhiro Onoe Hitoshi Tada Katsunori Tatsugami Takashi Ando Genki Kimura Kikuo Nomoto 《Cancer immunology, immunotherapy : CII》1998,47(4):198-204
A B16 melanoma-specific CD8+ T cell line (AB1) was established from the spleen cells of C57BL/6 mice cured of B16 melanoma with interleukin (IL)-12 treatment.
The AB1 line exclusively used T cell receptor Vβ11. The AB1 cells exhibited a cytolytic activity against both syngeneic B16 melanoma and allogeneic P815 mastocytoma, whereas
a cold inhibition assay revealed specificity of the AB1 cells against B16 melanoma. Their lostability to kill a class I loss
variant of B16 melanoma was restored by the transfection of H-2Kb gene. In addition, their interferon (IFN)-γ production was significantly suppressed by the addition of anti-H-2Kb monoclonal antibody, and RT-PCR analysis showed that the AB1 line expressed the mRNA encoding IFN-γ, but not IL-4 or IL-10.
The experiment using synthetic peptides of tyrosinase-related protein-2 (TRP-2) revealed that the AB1 cells could recognize
TRP-2181–188 peptide. Moreover, the AB1 cells showed an in vivo antitumor effect against established pulmonary metastases of B16 melanoma.
Overall, these results indicate that the Tc1-type Vβ11
+ AB1 cells exert an antitumor activity against syngeneic B16 melanoma through recognition of TRP-2181–188 peptide in an H-2Kb-restricted manner.
Received: 4 June 1998 / Accepted: 21 July 1998 相似文献
73.
Yoichi Moroi Yasuhiro Koga Kazuhiko Nakamura Masumi Ohtsu Genki Kimura Kikuo Nomoto 《Microbiology and immunology》1993,37(5):369-381
The role of lck gene in T cell proliferation and differentiation was investigated with transgenic mice carrying human lck cDNA whose expression was regulated by the promoter of mouse H-2Kb and the enhancer element of mouse IgH. RNase protection assay revealed that the lck transgene was expressed in the thymus and spleen, whereas immunoblot analysis demonstrated that amounts of p56lck in freshly isolated lymphoid organs were almost equal between transgenic mice and negative littermates. Cell-surface marker analyses of the thymocytes and peripheral lymphocytes revealed no remarkable difference between both groups. Notable finding is that the thymocytes from transgenic mice showed a significant proliferative response to the stimulation with IL-2, but not the thymocytes from negative littermates. Further analysis revealed that CD4+8– single positive thymocytes proliferated in response to IL-2. While surface expression levels of IL-2Rα and IL-2Rβ of these CD4+8– thymocytes from transgenic and control mice were almost equal before stimulation with IL-2, the expression of IL-2Rβ was induced only in transgenic thymocytes after stimulation with IL-2. Immunoblot analysis demonstrated that the expression of p56lck of transgenic thymocytes was not down-reguated at 4 hr after stimulaion with IL-2, whereas p56lck of control ones were not detectable any more at 4 hr after stimulation with IL-2. Moreover, in vitro kinase assay substantiated such unchanged expression of p56lck in the thymocytes from transgenic mice: the kinase activities of p56lck did not decrease in thymocytes from transgenic mice after stimulation with IL-2, while kinase activities of control ones were significantly down-regulated by stimulation of IL-2. These results suggested that a significant proliferative response found in the thymocytes from lck-transgenic mice after the stimulation with IL-2 was caused by a constitutive expression of p56lck in these thymocytes even after the stimulation. Our findings, therefore, support a possibility that p56lck may play a role in the IL-2R-mediated signaling system in CD4+8– thymocytes. 相似文献
74.
Masatoshi Togami Kosei Yasumoto Tokujiro Yano Teruyoshi Ishida Genki Kimura Keizo Sugimachi Kikuo Nomoto 《Cytotechnology》1991,6(1):39-47
We examined a serum-free medium (designated as TYI 101) for the generation of lymphokine-activated killer (LAK) cells from human lymphocytes, regional lymph node lymphocytes (RLNL) and peripheral blood lymphocytes (PBL). TYI 101 medium consisted of, in addition to nutrient mixture, transferrin, insulin, fetuin, sodium selenite, 2-mercaptoethanol, o-phosphorylethanolamine, chick egg yolk and porcine kidney extract. These hormones were effective for supporting RLNL proliferation as assessed by (3H)-thymidine uptake. When human lymphocytes from two different sources were cultivated with recombinant interleukin 2 (rIL-2) in TYI 101 medium, LAK activity was generated. In cultures of PBL from a healthy donor, LAK cells were generated in TYI 101 medium as efficiently as in RPMI 1640 medium supplemented with 10% human AB-type serum (RPMI-AB). In cultures of RLNL from lung cancer patients, LAK activity obtained in TYI 101 medium was about sixty-five percent of that in RPMI-AB. However, the addition of a small amount of AB-type serum improved the generation of LAK activity, LAK cell expansion, and cell viability in TYI 101 medium. We conclude that TYI 101 medium can be used for the generation of LAK cells from human lymph node lymphocytes with supplementation of none or only a reduced amount of human serum.Abbreviations IMDM
Iscove's Modification of Dulbecco's Medium
- rIL-2
recombinant Interleukin
- LAK
Lymphokine-Activated Killer
- RLNL
Regional Lymph Node Lymphocytes
- PBL
Pheripheral Blood Lymphocytes
- PBS
Phosphate-Buffered Saline
- RBC
Red Blood Cells
- RPMI-AB
RPMI 1640 medium supplemented with 10% human AB-type serum
Address for offprints: Tsukuba Research Laboratory, Japan Synthetic Rubber Co., Ltd., 25 Miyukigaoka, Tsukuba-shi, Ibaraki, 305 Japan 相似文献
75.
Dr. Tokujiro Yano Teruyoshi Ishida Ichiro Yoshino Mitsuhiro Murata Kosei Yasumoto Genki Kimura Kikuo Nomoto Keizo Sugimachi 《Biotherapy》1991,3(3):245-251
We designed a unique regimen of adoptive immunotherapy with lymphokine-activated killer (LAK) cells and recombinant interleukin 2 (rIL-2) for application with surgical adjuvant therapy of cancer. The regimen features the prolonged (6 consecutive days) s.c. administration of low-dose rIL-2 and the transfer ofex vivo generated LAK cells from regional lymph node lymphocytes, obtained at the time of surgical operation. According to this regimen, 5 patients with primary lung cancer received immunotherapy about 2 weeks after surgery (pulmonary lobectomy). Clinical toxicities included fever(5/5), fatigue(5/5), slight(< 5%) weight gain(5/5), increase of pleural effusion at the lobectomy site(2/5), and edema formation(1/5). All toxicities reversed within 4 days after the completion of therapy. Rebound lymphocytosis after therapy ranged from 2.4 to 5.5-fold (mean, 4.3-fold) over the baseline. Peripheral blood lymphocytes obtained during this lymphocytosis exhibitedin vitro LAK activity in 4 of 5 patients. Thus, the regimen is considered to be well-tolerable and immunologically active in regard to the postoperative state of the patients. 相似文献
76.
Vertical distribution of organic constituents, i.e. total organic carbon (TOC), extractable organic carbon with ethyl acetate (EOC), hydrocarbons, phytol, sterols, fatty acids and phenolic acids in Lake Vanda was studied to elucidate their features in relation to the stratification of lake water and the distribution of lake organisms. The concentrations of TOC, EOC and sterols increased with depth and attained the maximum values of 25 and 1.5 mgC l–1 and 1.4 g l–1 in the bottom, respectively, while those of fatty acids showed the maximum value of 61 g l–1 at a depth of 55.4 m, along with the highest value of the ratio of unsaturated (UC16, uC18) to saturated (C16, C18) acids (8.5) and with the highest carbon preference index (35). Hydrocarbons were only found in the bottom layers (60.4 and 65.9 m) and bottom sediment. These results suggest strongly that the vertical distribution of lake organisms and their activity are quite different due to depth. In the bottom warm anoxic layers the degradation of organic materials must have occurred significantly and thus refractory organic materials should be concentrated. 相似文献
77.
Natori Y Nanamiya H Akanuma G Kosono S Kudo T Ochi K Kawamura F 《Molecular microbiology》2007,63(1):294-307
As zinc is an essential trace metal ion for all living cells, cells elaborate a variety of strategies to cope with zinc starvation. In Bacillus subtilis, genes encoding ribosomal proteins L31 and S14 are duplicated into two types: one type contains a zinc-binding motif (RpmE or RpsN), whereas the other does not (YtiA or YhzA). We have previously shown that displacement of RpmE (L31) by YtiA from already assembled ribosomes is controlled by zinc, and this replacement could contribute to zinc mobilization under zinc-limiting conditions. We propose here that the switch between the two types of S14 has a different significance. rpsN is indispensable for growth and depletion of RpsN results in defective 30S subunits. YhzA can functionally replace RpsN to allow continued ribosome assembly under zinc-limiting conditions. Unlike YtiA, YhzA appeared in the ribosome at a slower rate consistent with incorporation into newly synthesized, rather than pre-existing ribosomes. These results raise the possibility that YhzA is involved in a fail-safe system for the de novo synthesis of ribosomes under zinc-limiting conditions. 相似文献
78.
Toru Sugiyama Genki Hasegawa Chie Niikura Keiko Kuwata Yasutada Imamura Yosuke Demizu Masaaki Kurihara Atsushi Kittaka 《Bioorganic & medicinal chemistry letters》2017,27(15):3337-3341
Here we report the synthesis of new PNA monomers for pseudocomplementary PNA (pcPNA) that are fully compatible with standard Fmoc chemistry. The thiocarbonyl group of the 2-thiouracil (sU) monomer was protected with the 4-methoxy-2-methybenzyl group (MMPM), while the exocyclic amino groups of diaminopurine (D) were protected with Boc groups. The newly synthesized monomers were incorporated into a 10-mer PNA oligomer using standard Fmoc chemistry for solid-phase synthesis. Oligomerization proceeded smoothly and the HPLC and MALDI-TOF MS analyses indicated that there was no remaining MMPM on the sU nucleobase. The new PNA monomers reported here would facilitate a wide range of applications, such as antigene PNAs and DNA nanotechnologies. 相似文献
79.
Shinjiro Hamano Hiroki Yoshida Hiroaki Takimoto Koh-hei Sonoda Kazuhiro Osada Xiangdong He Yoichi Minamishima Genki Kimura Kikuo Nomoto 《Microbiology and immunology》1998,42(9):607-616
It has been recognized that macrophages play an important role in controlling virus infection in experimental animal models. To evaluate the role of macrophages in acute murine cytomegalovirus infection, macrophages in the spleen and the liver were eliminated by an intravenous injection of liposomes containing a cytolytic agent, dichloromethylene diphosphonate. The depletion of macrophages led to a significant increase of virus titer in the spleen and lungs in both susceptible BALB/c and resistant C57BL/6 mice during the first three days after intravenous infection. In the spleen, the increase of virus titer in macrophage-depleted BALB/c mice was much greater than that in NK cell-depleted mice. These results suggest that macrophages contribute to protection mainly by the mechanisms which are independent of NK cells during the first three days after infection. The increase of virus titer in macrophage-depleted C57BL/6 mice was as great as that in NK cell-depleted mice because of the high contribution of NK cells to protection in C57BL/6 mice. In the liver in both strains of mice, the effects of macrophage depletion on virus titer were not as much as those in the spleen and lungs. Furthermore, the local depletion of peritoneal macrophages resulted in a great increase of virus titer in the spleen at three days after intraperitoneal infection. We conclude that macrophages greatly contribute to decreasing the virus load in some organs possibly through either or both intrinsic and extrinsic mechanisms in the early phase of primary infection with murine cytomegalovirus. 相似文献
80.
Hydrobiologia - In Japan, numerous artificial dams constructed for erosion control or hydroelectric power generation have affected almost all rivers and resulted in isolation and fragmentation of... 相似文献