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241.
Akihiro Yoshida Kuniaki Takata Toshiko Kasahara Toshio Aoyagi Shozo Saito Hiroshi Hirano 《The Histochemical journal》1995,27(5):420-426
Summary Glucose is actively absorbed in the intestine by the action of the Na+-dependent glucose transporter. Using an antibody against the rabbit intestinal Na+-dependent glucose transporter (SGLT1), we examined the localization of SGLT1 immunohistochemically along the rat digestive
tract (oesophagus, stomach, duodenum, jejunum, ileum, colon and rectum). SGLT1 was detected in the small intestine (duodenum,
jejunum and ileum), but not in the oesophagus, stomach, colon or rectum. SGLT1 was localized at the brush border of the absorptive
epithelium cells in the small intestine. Electron microscopical examination showed that SGLT1 was localized at the apical
plasma membrane of the absorptive epithelial cells. SGLT1 was not detected at the basolateral plasma membrane. Along the crypt-villus
axis, all the absorptive epithelial cells in the villus were positive for SGLT1, whose amount increased from the bottom of
the villus to its tip. On the other hand, cells in the crypts exhibited little or no staining for SGLT1. Goblet cells scattered
throughout the intestinal epithelium were negative for SGLT1. These observations show that SGLT1 is specific to the apical
plasma membrane of differentiated absorptive epithelial cells in the small intestine, and suggest that active uptake of glucose
occurs mainly in the absorptive epithelial cells in the small intestine. 相似文献
242.
Hiroshi Nakada Mizue Inoue Nobuhiro Tanaka Nobutaka Wakamiya Ikuo Yamashina 《Glycoconjugate journal》1995,12(3):356-359
We have measured glycosyltransferase activities of SupT1 cells, a T-lymphoid cell line shown to react with autoantibodies in the sera of many HIV patients. Since considerable -N-acetylgalactosaminyl-transferase and 1, 3 galactosyl-transferase activities were found in SupT1 cells, at least the O-glycan core 1 structure can probably be synthesized. FACS analysis using an anti-T monoclonal antibody showed expression of the T antigen (Gal 1-3 GalNAc). Glycoproteins with the T antigen were isolated by immunoprecipitation with the anti-T antibody from a SupT1 cell lysate labelled metabolically with3H-glucosamine and then analysed by SDS-PAGE. It was revealed that the precipitate contained a glycoprotein with a molecular weight corresponding to that of leukosialin. O-glycans were prepared from the immunoprecipitate by alkaline-borohydride treatment and then fractionated on Bio-Gel P-2, GalNAcOH and Gal-GalNAcOH being identifiedinter alia. These results suggest that an anti-T antibody may be included in the autoantibodies found in HIV-1 infected individuals. 相似文献
243.
Aruto Yoshida Tomoka Hara Hiroshi Ikenaga Makoto Takeuchi 《Glycoconjugate journal》1995,12(6):824-828
By employing a bovine UDP-N-acetylgalactosamine: polypeptideN-acetylgalactosaminyl transferase (O-GalNAc transferase) cDNA as a probe, we isolated four overlapping cDNAs from a porcine lung cDNA library. Both the nucleotide sequence of the porcine cDNA and the predicted primary structure of the protein (559 amino acids) proved to be very similar to those of the bovine enzyme (95% and 99% identity, respectively). Transient expression of the clone in COS-7 cells, followed by enzymatic activity assays, demonstrated that this cDNA sequence encodes a porcine O-GalNAc transferase. The intracellular O-GalNAc transferase activity was increased approximately 100-fold by transfecting cells with the porcine cDNA.Abbreviations O-GalNAc transferase
UDP-N-acetylgalactosamine: polypeptideN-acetylgalactosaminyltransferase
- PCR
polymerase chain reaction
- SDS
sodium dodecyl sulfate
- PAGE
polyacrylamide gel electrophoresis
- GnT-III
UDP-N-acetylglucosamine: -mannoside -1,4N-acetylglucosaminyltransferase III 相似文献
244.
The authors established apoptosis resistant COS–1, myeloma, hybridoma, and Friend leukemia cell lines by genetically engineering cells, aiming at more efficient protein production by cell culture. COS–1 cells, which are most widely used for eukariotic gene expression, were transfected with human bcl–2 gene. Both bcl–2 and mock transfected COS–1 cells were cultured at low (0.2%) serum concentration for 9 days. The final viable cell number of the bcl–2 transfected cells was ninefold of that of the mock transfectants. Both bcl–2 and mock transfectants were further transfected with the vector pcDNA- containing SV40 ori and immunoglobulin gene for transiently expressing protein. The bcl–2 expressing COS–1 cells produced more protein than the mock transfected COS–1 cells after 4 days posttransfection.Mouse myeloma p3-X63-Ag.8.653 cells, which are widely used as the partner for preparing hybridoma, and hybridoma 2E3 cells were transfected with human bcl–2 gene. Both bcl–2 transfected myeloma and hybridoma survived longer than the corresponding original cells in batch culture. The bcl–2 transfected 2E3 cells survived 2 to 4 four days longer in culture, producing 1.5- to 4-fold amount of antibody in comparison with the mock transfectants.Coexpression of bag–1 with bcl–2 improved survival of hybridoma 2E3 cells more than bcl–2 expression alone. The bag–1 and bcl–2 coexpressing cells produced more IgG than the the cells expressing bcl–2 alone.Apoptosis of Friend murine erythroleukemia(F-MEL) cells was suppressed with antisense c-jun expression. The antisense c-jun expressing cells survived 16 days at non-growth state. 相似文献
245.
246.
Kazumasa Tsuji Masaharu Iijima Hiroshi Matsuzawa Shinji Sakamoto 《Biotechnology Techniques》1997,11(6):395-398
Escherichia coli JM109(DE3) harboring expression plasmid pkAQNÆC30, which carries the Thermus protease aqualysin I (AQI) gene, was cultivated with glucose as a sole carbon source. The final cell concentration was over 15 g dry weight/l and the amount of AQI produced reached approximately 130 kU/ml broth. Moreover, by using two carbon sources, glucose and glycerol, the production yield was increased to over 200 kU AQI/ml, while suppressing the formation of inhibitory acetic acid. 相似文献
247.
Unique tissue distribution of a mouse macrophage C-type lectin 总被引:7,自引:2,他引:5
Mizuochi Shigeki; Akimoto Yoshihiro; Imai Yasuyuki; Hirano Hiroshi; Irimura Tatsuro 《Glycobiology》1997,7(1):137-146
We examined mouse tissue for the expression of macrophage galactose/N-acetylgalactosamine-specificC-type lectin using a rat monoclonal antibody (mAb) specificfor this lectin (mAb LOM-14). The binding of mAb LOM-14 wasdetected in detergent extracts from tissue by means of immunoblottinganalysis. It was shown that this mAb did not cross-react withmouse hepatic lectins, a structural homologue. The macrophagelectin was widely distributed among various mouse tissues asjudged by the affinity isolation followed by the immunochemicaldetection. The exceptions were brain, liver, kidney, small intestine,and peripheral blood. Extracts from these organs exhibited,at best, very weak signals upon mAb LOM-14 binding, despitethe presence of cells expressing macrophage markers. The mostintense signal was observed in the extract from skin, suggestingthat cells expressing this lectin are abundant in skin. Thetissues shown to contain this lectin were further investigatedby immunohistochemical staining of the sections. Cells weredistributed in the connective tissue and in the interstice,particularly the dermis and subcutaneous layer of skin. Cellslocalized in the epithelium of skin (epidermis) or other epitheliathat we examined were not stained. Perivascular localizationof cells stained with mAb LOM-14 was also demonstrated in cardiacand skeletal muscle tissues. Immunoelectron microscopy revealedthe presence of this lectin along the rough endoplasmic reticulum.In conclusion, the distribution of C-type lectin specific forgalactose/N-acetylgalactosamine in mice was unique. The connectivetissue-specific distribution should provide important informationon the biological role of this lectin. lectin macrophage calcium-type lectin connective tissue 相似文献
248.
Physical effects of negative air ions in a wet sauna 总被引:10,自引:0,他引:10
I. Watanabe Hiroshi Noro Yoshinori Ohtsuka Yukio Mano Yuko Agishi 《International journal of biometeorology》1997,40(2):107-112
The physical effects of negative air ions on humans were determined in an experimental sauna room equipped with an ionizer.
Thirteen healthy persons took a wet sauna bath (dry bulb temperature 42° C, relative humidity 100%, 10 min exposure) with
or without negative air ions. The subjects were not told when they were being exposed to negative air ions. There were no
differences in the moods of these persons or changes in their blood pressures between the two saunas. The surface temperatures
of the foreheads, hands, and legs in the sauna with negative ions were significantly higher than those in the sauna without
ions. The pulse rates and sweat produced in the sauna with ions were singificantly higher than those in the sauna without
ions. The results suggest that negative ions may amplify the effects on humans of the sauna.
Received: 31 March 1995 / Revised: 25 July 1995 / Accepted: 26 July 1996 相似文献
249.
Reproduction and development of Pratylenchus penetrans were studied on genetically transformed ladino clover roots. Solitary females developing on transformed roots in nutrient gellan gum medium (pH 5.5) deposited 1.2, 1.5, 1.6, 1.8, and 2.0 eggs per day at the respective temperatures of 17, 20, 25, 27, and 30 °C. The number of eggs deposited was highly correlated with temperature. A reduction in egg-laying rates at the start of hatching was observed at all temperatures. Juvenile mortality was higher at 17 °C (50.4%), 20 °C (50.3%), and 30 °C (58.4%) than at 25 °C (34.6%) and 27 °C (37.6%). Life-cycle (egg deposition to egg deposition) duration was 46, 38, 28, 26, and 22 days at the respective temperatures. The developmental zero degrees (°C) and the effective accumulative temperatures (degree-days) required for hatching, female emergence, and onset of oviposition (completion of one generation) of P. penetrans were estimated to be 2.7 and 200, 4.2 and 548, and 5.1 and 564, respectively. Pratylenchus penetrans reproduces over a wide range of temperatures. 相似文献
250.
Abstract: It has been previously reported that Alzheimer's amyloid β protein (Aβ) induces reactive astrocytosis in culture. In the present study, we found that Aβ potently inhibits cellular redox activity of cultured astrocytes, as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay. The following comparative studies revealed several differences between these two actions of Aβ on astrocytes. First, Aβ-induced reactive morphological change was suppressed by the presence of serum or thrombin, and Aβ inhibition of cellular redox activity was observed in either the presence or the absence of serum. Second, micromolar concentrations (10 µ M or more) were required for Aβ to induce reactive astrocytosis, whereas nanomolar concentrations (0.1–100 n M ) were sufficient to inhibit cellular redox activity. Third, the effect of micromolar Aβ was virtually irreversible, but nanomolar Aβ-induced inhibition of cellular redox activity was reversed by washing out Aβ. Furthermore, as it has been reported that Aβ neurotoxicity is mediated by reactive oxygen species, we also examined if similar mechanisms are involved in astrocytic response to Aβ. However, neither Aβ-induced morphological change nor inhibition of redox activity was blocked by antioxidants, suggesting that these effects are not caused by oxidative stress. 相似文献