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71.
High-molecular-weight kininogen is exclusively membrane bound on endothelial cells to influence activation of vascular endothelium 总被引:3,自引:1,他引:3
An important biologic function of high-molecular-weight kininogen (HK) is to deliver bradykinin (BK) to its cellular receptors. Internalization and degradation of HK may provide a mechanism by which endothelial cells modulate the production of BK and control its activities. Therefore, we investigated the binding and subsequent distribution of biotinylated-HK (biotin-HK) associated with human umbilical vein endothelial cells (HUVEC). HUVEC bound 3 to 4 times more HK and with greater avidity at 1 to 3 hours at 37 degrees C than at 4 degrees C (Bmax = 1.0 +/- 0.02 x 10(7) molecules/cell, kd = 7 +/- 3 nmol/L v Bmax = 2.6 +/- 0.2 x 10(6) molecules/cell, kd = 46 +/- 8 nmol/L). However, there was no evidence that the difference was caused by internalization of HK at the higher temperature. First, the same amount of biotin-HK was associated with nonpermeabilized and permeabilized HUVEC using buffers containing 20 to 50 mumol/L zinc ion in the absence or presence of 2 mmol/L calcium ion. Second, binding of biotin-HK to HUVEC was approximately 92% reversible at 1 hour when the cells were maintained at both 37 degrees C and 4 degrees C. Third, neither chloroquine nor primaquine altered the amount of biotin-HK bound to HUVEC. Fourth, biotin-HK bound to HUVEC was almost completely removed by pronase. Fifth, the nonpermeable dye, crystal violet, almost completely quenched the fluorescence signal emitted by HUVEC-associated fluorescein isothiocyanate (FITC) HK. Finally, the localization of HUVEC-bound FITC-HK was restricted to the membrane as shown by laser scanning confocal microscopy. The expression of HK binding sites had an absolute requirement for metabolic energy, but was not dependent on new protein synthesis. Membrane-bound HK contributed to the anticoagulant nature of endothelial cells by blocking human alpha-thrombin binding and its resultant induction of prostacyclin formation. These studies indicate that HK is not internalized by HUVEC, but remains primarily on cell surfaces to be accessible for BK liberation and to modulate the binding and actions of alpha-thrombin. 相似文献
72.
Role of light chain variable region in myeloma with light chain deposition disease: evidence from an experimental model 总被引:4,自引:2,他引:4
Khamlichi AA; Rocca A; Touchard G; Aucouturier P; Preud'homme JL; Cogne M 《Blood》1995,86(10):3655-3659
Light chain deposition disease (LCDD) results from a propensity of some human monoclonal L chains to form tissue deposits. We designed an experimental model for in vivo expression of human kappa L chain sequences in mice and compared a somatically mutated LCDD chain with a closely related control kappa chain, both encoded by the unique V kappa IV gene. Mice secreting the LCDD chain but not those producing the control chain showed deposits with a distribution similar to that observed in patients. These data show that discrete changes in V region sequences can play a major role in tissue deposition of human L chains. 相似文献
73.
The Ig heavy chain (IgH) gene was used as a marker to investigate clonal succession and the origin of the neoplastic cell in multiple myeloma. The polymerase chain reaction (PCR) was used to amplify a section of the rearranged IgH gene at diagnosis and at progression in 21 patients who had exhibited a plateau phase. A monoclonal PCR product was seen for 16 of the patients and the product present at progression was of the same molecular weight as that at diagnosis. This finding suggests that the IgH rearrangement present at diagnosis and progression was the same. This was confirmed by sequencing the IgH gene in 10 patients. The IgH genes were found to be hypermutated at diagnosis, but no further hypermutation occurred during the course of the disease. The results provide evidence that the neoplastic cell in myeloma may originate as a memory B cell, plasmablast, or plasma cell, and suggest that progression beyond the plateau phase is not caused by clonal succession. 相似文献
74.
Heterogeneity of breakpoints of 11q23 rearrangements in hematologic malignancies identified with fluorescence in situ hybridization 总被引:3,自引:0,他引:3
Kobayashi H; Espinosa R d; Thirman MJ; Gill HJ; Fernald AA; Diaz MO; Le Beau MM; Rowley JD 《Blood》1993,82(2):547-551
Twenty-four patients whose cells contained a variety of 11q23 rearrangements, including translocations, insertions, and an inversion, were studied using fluorescence in situ hybridization with cosmid, phage, and plasmid probes mapped to 11q22-24. In 17 patients, the breakpoints of the common 11q23 translocations involving chromosomes 4, 6, 9, and 19 as well as some uncommon translocations involving 3q23, 17q25, 10p11, and an insertion 10;11 were all located in the breakpoint cluster region of the MLL gene, regardless of age, phenotype of disease, or involvement of a third chromosome. The breakpoints in 11q23 in the other 7 patients with a t(7;11)(p15;q23), inv(11)(p11q23), t(4;11)(q23;q23), der(5)t(5;11)(q13;q23), ins(10;11)(p11;q23q24), t(11;14)(q23;q11), or t(11;18;11) (p15;q21;q23) were located either centromeric to CD3D or telomeric to THY1. Thus, although most 11q23 rearrangements, involve the same breakpoint cluster region of MLL, there is heterogeneity in the breakpoint in some of the rare rearrangements. 相似文献
75.
Digital and conventional chest images: observer performance with Film Digital Radiography System 总被引:5,自引:0,他引:5
The Film Digital Radiography System (FilmDRS) is a device with a laser optical film digitizer, 2,000 X 2,000 X 12-bit memory, and a 1,000-line video display. To evaluate the adequacy of this device for general radiography of the chest, four readers independently analyzed both radiographs and the corresponding video display of the digitized chest images of 150 patients, consisting of 100 images of abnormalities and 50 normal images. The overall results indicate equal sensitivity for the two systems. The FilmDRS, with interactive windowing, proved superior in the detection of hilar and mediastinal disease. X-ray film was superior in allowing detection of hyperlucent states. There was equivalent sensitivity for other disease categories. Superior specificity was achieved with conventional radiographs. 相似文献
76.
77.
78.
Okada AA; Keino H; Suzuki J; Sakai J; Usui M; Mizuguchi J 《International immunology》1998,10(12):1917-1922
The systemic administration of IFN-alpha/beta was previously found to
suppress inflammation in rats with experimental autoimmune uveoretinitis
(EAU); however, an effect on the systemic immune response was not
identified. In order to investigate an immunological basis for suppression
at the intraocular level, rats immunized with interphotoreceptor
retinoid-binding protein (IRBP) were administered daily intramuscular
injections of 10(5) IU IFN-alpha/beta and cytokines were measured by ELISA
in intraocular extracts prepared by ultrasonification at various timepoints
throughout the course of EAU. In control EAU, intraocular concentrations of
IFN-gamma were found to be non-detectable on day 8 before the onset of
inflammation, significantly elevated on day 12 at peak inflammation
(182+/-106 pg/ml), then non-detectable again on day 16 after inflammation
had begun to subside. In contrast, intraocular IFN-gamma in IFN-alpha/beta-
treated rats remained non-detectable or low at all timepoints. Measurement
of intraocular IL-2 revealed no difference between the two groups of rats.
Intraocular IL-4 concentrations were elevated in rats treated with
IFN-alpha/beta, although this cytokine was also detected in the same range
in controls as well as normal rats. Finally, intraocular IL-10 was
non-detectable on day 8, significantly elevated at peak inflammation on day
12 (588+/-139 pg/ml), then decreased to low levels on day 16 in control EAU
rats, while remaining non-detectable or low in IFN-alpha/beta-treated rats.
These results suggest that acute inflammation in IRBP-induced EAU in rats
involves both IFN-gamma and IL- 10 at the local intraocular level, and that
systemic administration of IFN-alpha/beta inhibits EAU via a mechanism that
involves suppression of both cytokines.
相似文献
79.
Stéphanie F. Bernatchez PhD ; Patrick J. Parks MD PhD ; Duane M. Grussing SRS ; Shawn L. Matalas CVT ; Gwen S. Nelson AA 《Wound repair and regeneration》1998,6(3):223-233
Chronic wounds, such as venous ulcers and pressure ulcers, frequently remain unresponsive to currently available treatments. Several animal models of wound healing have been published, including models of impaired healing developed to mimic the clinical condition of chronic wounds better. We used a delayed wound healing model in the pig that uses irradiation of the skin prior to creation of the surgical wounds and characterized it histologically. Radiation was used on one side of the back prior to making four full-thickness wounds on each side. Clinical observations were performed to record granulation tissue, reepithelialization, and wound area as a function of time. Histology data were obtained at 1, 2, 3, and 4 weeks, and slides were stained with hematoxylin and eosin for general observations. Immunohistochemistry was performed using laminin as a marker for blood vessels, and the number, size, and circularity of blood vessels found in the granulation tissue were measured. Our results show that this model causes a delay in wound healing that is mostly apparent between days 7 and 15. Granulation tissue took more time to form and fill the wounds on the irradiated side, and blood vessels were slower to develop. Blood vessels were larger and more irregular in shape on the irradiated side than on the control side. After 2 weeks, healing resumed, indicating that the induced damage was not irreversible. These results suggest that this model can be used to test the effects of therapeutic approaches intended to treat chronic wounds. 相似文献
80.