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排序方式: 共有297条查询结果,搜索用时 15 毫秒
31.
Anne C Fischer Carolina I Smith Liudmila Cebotaru Xuemei Zhang Frederic B Askin Jerry Wright Sandra E Guggino Robert J Adams Terence Flotte William B Guggino 《Molecular therapy》2007,15(4):756-763
Gene therapy using recombinant adeno-associated virus (rAAV2) vectors for cystic fibrosis has shown gene transfer and remarkable safety, yet indeterminate expression. A new construct has been characterized with a powerful exogenous promoter, the cytomegalovirus enhancer/chicken beta-actin promoter, driving a truncated CF transmembrane conductance regulator (CFTR), pseudotyped in an AAV5 viral coat. Our goal is to demonstrate that airway delivery of a pseudotyped rAAV5 vector results in gene transfer as well as expression in non-human primates. Aerosolized pseudotyped rAAV5-DeltaCFTR or rAAV5-GFP (green fluorescent protein) genes were delivered to four and six lungs, respectively. The pseudotyped rAAV5 vector did result in GFP gene transfer (1.005x10(6) copies/mug DNA on average) and quantifiable gene expression. Microscopy confirmed protein expression in airway epithelium. Similarly, the vector also resulted in vector-specific CFTR DNA (1.24x10(5) copies/microg) and mRNA expression. Immunoprecipitation and (32)P phosphoimaging were used to demonstrate CFTR protein expression, as qualitatively enhanced beyond the barely detectable endogenous expression in untreated animals. Based on these promising studies, this CFTR minigene construct is a therapeutic candidate. 相似文献
32.
33.
Hinsch E; Ponce AA; Hagele W; Hedrich F; Muller-Schlosser F; Schill WB; Hinsch KD 《Human reproduction (Oxford, England)》1997,12(8):1673-1681
Binding of mammalian spermatozoa to the zona pellucida and the induction of
the acrosome reaction are prerequisites for successful oocyte
fertilization. It has been postulated that xenobiotics that are released in
the environment as well as exposure to pharmaceutical medications may be
associated with reproductive problems in men and wildlife. Examining
physiological and non-physiological effects of particular compounds on
sperm functions requires high quality in-vitro test systems. We established
a reliable combined in-vitro test system with bovine gametes and evaluated
if aliquots of pooled post-thaw spermatozoa are suitable for examining
essential sperm functions. Using cryopreserved semen, the PSA-FITC/Hoechst
33258 staining procedure was applicable to evaluate the acrosomal status
and cell viability. In the bovine hemizona assay, hemizona indices revealed
no differences between cryopreserved and fresh semen. Treatment of
post-thaw bovine spermatozoa with progesterone (1 microM or bovine
follicular fluid (20%) induced the acrosome reaction from 12% (untreated
spermatozoa) to 25% (P < 0.001) and to 22% [corrected] (P < 0.01),
respectively. Incubation of both compounds (1 microM progesterone and 20%
follicular fluid) raised the percentage of acrosome-reacted spermatozoa to
30% (P < 0001). Our results demonstrate that cryopreserved semen can be
integrated into an in-vitro screening model for reproductive toxicology
testing. Pooled, cryopreserved bovine spermatozoa will thus permit
reproducible experiments for clinical and basic science purposes and may
also be applicable for the human system.
相似文献
34.
35.
H‐ferritin and proinflammatory cytokines are increased in the bone marrow of patients affected by macrophage activation syndrome
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P. Ruscitti P. Cipriani P. Di Benedetto V. Liakouli O. Berardicurti F. Carubbi F. Ciccia G. Guggino G. Triolo R. Giacomelli 《Clinical and experimental immunology》2018,191(2):220-228
Macrophage activation syndrome (MAS) is hyperinflammatory life‐threatening syndrome, associated typically with high levels of serum ferritin. This is an iron storage protein including heavy (H) and light (L) subunits, categorized on their molecular weight. The H‐/L subunits ratio may be different in tissues, depending on the specific tissue and pathophysiological status. In this study, we analysed the bone marrow (BM) biopsies of adult MAS patients to assess the presence of: (i) H‐ferritin and L‐ferritin; (ii) CD68+/H‐ferritin+ and CD68+/L‐ferritin+; and (iii) interleukin (IL)‐1β, tumour necrosis factor (TNF) and interferon (IFN)‐γ. We also explored possible correlations of these results with clinical data. H‐ferritin, IL‐1β, TNF and IFN‐γ were increased significantly in MAS. Furthermore, an increased number of CD68+/H‐ferritin+ cells and an infiltrate of cells co‐expressing H‐ferritin and IL‐12, suggesting an infiltrate of M1 macrophages, were observed. H‐ferritin levels and CD68+/H‐ferritin+ cells were correlated with haematological involvement of the disease, serum ferritin and C‐reactive protein. L‐ferritin and CD68+/L‐ferritin+ cells did not correlate with these parameters. In conclusion, during MAS, H‐ferritin, CD68+/H‐ferritin+ cells and proinflammatory cytokines were increased significantly in the BM inflammatory infiltrate, pointing out a possible vicious pathogenic loop. To date, H‐ferritin and CD68+/H‐ferritin+ were associated significantly with haematological involvement of the disease, suggesting biomarkers assessing severity of clinical picture. 相似文献
36.
Caterina Maria Gambino Danilo Di Bona Anna Aiello Ciriaco Carru Giovanni Duro Giuliana Guggino Angelo Ferrante Angelo Zinellu Calogero Caruso Giuseppina Candore Giulia Accardi 《Human immunology》2018,79(3):172-177
Recently, the role of killer cell immunoglobulin-like receptor (KIR) in autoimmune diseases has received increasing attention. The present study was undertaken to determine the association of KIR genes and the human leukocytes antigen (HLA) ligands with Systemic Lupus Erythematosus (SLE) and accompanying oxidative stress. Presence or absence of 17 KIR and 5 HLA loci was performed using the polymerase chain reaction-sequence specific primer (PCR-SSP) method by case-control study. A total of 45 SLE patients, and 60 healthy controls, all of Sicilian descent, were enrolled. Plasma values of the anti-oxidant molecule Taurine were determined in all subjects by capillary electrophoresis UV detection. The carrier frequency of the KIR2DS2 gene was significantly increased in SLE patients compared to healthy controls (73.3 versus 45.0%; OR?=?3.36; 95% CI?=?1.46–7.74; p?=?.005) suggesting a role of KIR2DS2 gene in the susceptibility to disease. We also observed a strong positive association between the presence of HLA-C1 ligands group and the disease (82.2% in SLE patients versus 41.7% in controls; OR?=?6.47, 95% CI?=?2.58–16.26; p?<?.0001). Stepwise logistic regression analysis supported the effect of the HLA-C1 ligands in SLE patients (OR?=?7.06, 95% CI?=?0.07–2.19; p?=?.002), while the KIR genes were no longer significant. Interestingly, we found that SLE patients HLA-C1 positive showed significantly decreased plasma levels of antioxidant activity marker Taurine (69.38?±?28.49?μmol/L) compared to SLE patients HLA-C1 negative (108.37?±?86.09?μmol/L) (p?=?.03). In conclusion, HLA-C1 ligands group was significantly associated with an increased risk of SLE as well as an increased oxidative stress status overall in SLE patients. 相似文献
37.
Cloning of the gamma-aminobutyric acid (GABA) rho 1 cDNA: a GABA receptor subunit highly expressed in the retina. 总被引:10,自引:6,他引:10
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38.
39.
Antonino Tuttolomondo Rosaria Pecoraro Alessandra Casuccio Domenico Di Raimondo Carmelo Buttà Giuseppe Clemente Vittoriano della Corte Giuliana Guggino Valentina Arnao Carlo Maida Irene Simonetta Rosario Maugeri Rosario Squatrito Antonio Pinto 《Medicine》2015,94(20)
CD4+ CD28− T cells also called CD28 null cells have been reported as increased in the clinical setting of acute coronary syndrome. Only 2 studies previously analyzed peripheral frequency of CD28 null cells in subjects with acute ischemic stroke but, to our knowledge, peripheral frequency of CD28 null cells in each TOAST subtype of ischemic stroke has never been evaluated. We hypothesized that CD4+ cells and, in particular, the CD28 null cell subset could show a different degree of peripheral percentage in subjects with acute ischemic stroke in relation to clinical subtype and severity of ischemic stroke.The aim of our study was to analyze peripheral frequency of CD28 null cells in subjects with acute ischemic stroke in relation to TOAST diagnostic subtype, and to evaluate their relationship with scores of clinical severity of acute ischemic stroke, and their predictive role in the diagnosis of acute ischemic stroke and diagnostic subtypeWe enrolled 98 consecutive subjects admitted to our recruitment wards with a diagnosis of ischemic stroke. As controls we enrolled 66 hospitalized patients without a diagnosis of acute ischemic stroke. Peripheral frequency of CD4+ and CD28 null cells has been evaluated with a FACS Calibur flow cytometer.Subjects with acute ischemic stroke had a significantly higher peripheral frequency of CD4+ cells and CD28 null cells compared to control subjects without acute ischemic stroke. Subjects with cardioembolic stroke had a significantly higher peripheral frequency of CD4+ cells and CD28 null cells compared to subjects with other TOAST subtypes. We observed a significant relationship between CD28 null cells peripheral percentage and Scandinavian Stroke Scale and NIHSS scores. ROC curve analysis showed that CD28 null cell percentage may be useful to differentiate between stroke subtypes.These findings seem suggest a possible role for a T-cell component also in acute ischemic stroke clinical setting showing a different peripheral frequency of CD28 null cells in relation of each TOAST subtype of stroke. 相似文献
40.
Overcoming the Cystic Fibrosis Sputum Barrier to Leading Adeno-associated Virus Gene Therapy Vectors
Benjamin S Schuster Anthony J Kim Joshua C Kays Mia M Kanzawa William B Guggino Michael P Boyle Steven M Rowe Nicholas Muzyczka Jung Soo Suk Justin Hanes 《Molecular therapy》2014,22(8):1484-1493
Gene therapy has not yet improved cystic fibrosis (CF) patient lung function in human trials, despite promising preclinical studies. In the human CF lung, inhaled gene vectors must penetrate the viscoelastic secretions coating the airways to reach target cells in the underlying epithelium. We investigated whether CF sputum acts as a barrier to leading adeno-associated virus (AAV) gene vectors, including AAV2, the only serotype tested in CF clinical trials, and AAV1, a leading candidate for future trials. Using multiple particle tracking, we found that sputum strongly impeded diffusion of AAV, regardless of serotype, by adhesive interactions and steric obstruction. Approximately 50% of AAV vectors diffused >1,000-fold more slowly in sputum than in water, with large patient-to-patient variation. We thus tested two strategies to improve AAV diffusion in sputum. We showed that an AAV2 mutant engineered to have reduced heparin binding diffused twice as fast as AAV2 on average, presumably because of reduced adhesion to sputum. We also discovered that the mucolytic N-acetylcysteine could markedly enhance AAV diffusion by altering the sputum microstructure. These studies underscore that sputum is a major barrier to CF gene delivery, and offer strategies for increasing AAV penetration through sputum to improve clinical outcomes. 相似文献