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991.
目的: 制备靶向表皮生长因子受体(epithelial growth factor receptor,EGFR)隐蔽表位(287-302)的免疫毒素,并鉴定其生物学功能。 方法: 通过基因工程方法将抗EGFR(287-302)的806单链抗体(806 single-chain antibody fragment, 806scFv )基因经柔性肽与铜绿假单胞菌外毒素A(Pseudomonas exotoxin A, PEA)的截短形式PE38KDEL连接,构建原核表达载体pET-22b-806scFv-PE38KDEL并转化至大肠杆菌BL21(DE3)中,经纯化获得该免疫毒素融合蛋白806scFv-PE38KDEL,ELISA和流式细胞术检测其与EGFR的结合活性,间接免疫荧光检测重组免疫毒素的内化作用,CCK-8法检测806scFv-PE38KDEL对人脑胶质瘤细胞U87MG和U87MG-EGFRvⅢ、表皮癌细胞A431、乳腺癌细胞MDA-MB-468、舌癌细胞CAL-27的细胞毒性。 结果: 成功构建重组免疫毒素806scFv-PE38KDEL,诱导表达的蛋白806scFv-PE38KDEL以包涵体形式存在,经纯化后的纯度>95%,经SDS-PAGE和Western blotting 鉴定为目的蛋白。806scFv-PE38KDEL 能EGFRvⅢ胞外段蛋白结合,还能与外源性表达EGFRvⅢ的肿瘤细胞和高表达EGFR的肿瘤细胞相结合,而与表达低水平EGFR的肿瘤细胞不结合。806scFv介导了重组免疫毒素的内化。806scFv-PE38KDEL对靶细胞有明显的杀伤作用,对过表达EGFRvⅢ的U87MG-EGFRvⅢ细胞IC50值为(5.85±0.03) ng/ml,对EGFR高表达细胞MDA-MB-468、A431、CAL-27的IC50值分别为(162.80±0.06)、(75.72±0.04)、(123.70±0.03) ng/ml。在1 μg/ml的质量浓度下,相比PBS对照组,806scFv-PE38KDEL对U87MG-EGFRvⅢ 、MDA-MB-468、A431和CAL-27细胞增殖的抑制率均显著增高\[(98.67±0.07)% vs (2.45±2.85)%、(86.26±101)% vs (0.48±1.76)%、(96.72±016)% vs (1.33±1.31)%、(96.29±0.30)% vs (2.00±0.60)%,均P<0.01\],而对U87MG细胞几乎没有抑制作用\[(359±2.09)% vs (0.19±0.95),P>0.05\]。 结论: 本研究所制备的靶向EGFR(287-302)表位的重组免疫毒素806scFv-PE38KDEL能特异地结合并杀伤EGFRvⅢ或EGFR高表达的肿瘤细胞。 相似文献
992.
胃底静脉曲张出血是肝硬化严重并发症,内镜下组织胶注射治疗已成为胃底静脉曲张的一线治疗,其异位栓塞等并发症也多次被报道。本篇报道了1例胃底静脉曲张急诊出血病例,行内镜下组织胶注射治疗后出现罕见的脑动脉空气栓塞。 相似文献
993.
A novel photo-Fenton catalytic system for the removal of organic pollutants was presented, including the use of photo-Fenton process and a submerged magnetic separation membrane photocatalytic reactor (SMSMPR). We synthesized TiO2–Fe3O4 composites as the photocatalyst and made full use of the magnetism of the photocatalyst to realize the recollection of the catalyst from the medium, which is critical to the commercialization of photocatalytic technology for wastewater treatment. The photo-Fenton performance of TiO2–Fe3O4 is evaluated with amoxicillin trihydrate (AMX) as a target pollutant. The results indicate that the TiO2–Fe3O4/H2O2 oxidation system shows efficient degradation of AMX. Fe3O4 could not only enhance the heterogeneous Fenton degradation of organic compounds but also allow the photocatalyst to be magnetically separated from treated water. After four reaction cycles, the TiO2–Fe3O4 composites still exhibit 85.2% removal efficiency of AMX and show excellent recovery properties. Accordingly, the SMSMPR with the TiO2–Fe3O4 composite is a promising way for removing organic pollutants.With a TiO2–Fe3O4 composite as the catalyst, amoxicillin was degraded via a photo-Fenton process using a submerged magnetic separation membrane photocatalytic reactor. 相似文献
994.
Wenbao Wang Linghua Kong Heyuan Zhao Ronghua Dong Jianjiang Li Zhanhua Jia Ning Ji Shucai Deng Zhiming Sun Jing Zhou 《European spine journal》2007,16(8):1119-1128
Thoracic ossification of ligamentum flavum (OLF) caused by skeletal fluorosis is rare. Only six patients had been reported in the English literature. This study reports findings from the first clinical series of this disease. This was a retrospective study of patients with thoracic OLF due to skeletal fluorosis who underwent surgical management at the authors' hospital between 1993 and 2003. Diagnosis of skeletal fluorosis was made based on the epidemic history, clinical symptoms, radiographic findings, and urinalysis. En bloc laminectomy decompression of the involved thoracic levels was performed in all cases. Cervical open door decompression or lumbar laminectomy decompression was performed if relevant stenosis was present. Neurological status was evaluated preoperatively, at the third day postoperatively, and at the end point of follow-up using the Japanese Orthopaedic Association (JOA) scoring system of motor function of the lower extremities. A total of 23 cases were enrolled, 16 (69.6%) males and 7 (30.4%) females, age ranging from 42 to 72 years (mean 54.8 years). All patients came from a high-fluoride area, and 22 (95.7%) had dental fluorosis. Medical imaging showed OLF together with ossification of many ligaments and interosseous membranes, including interosseous membranes of the forearm (18/23 patients 78.3%), leg (14/23 patients 60.9%), and ribs (11/23 patients 47.8%). OLF was classified into five types based on MRI findings: localized (4/23 patients 17.4%), continued (12/23 patients 52.2%), skip (3/23 patients 13.0%), combining with anterior pressure (2/23 patients 8.7%), and combining with cervical and/or lumbar stenosis (2/23 patients, 8.7%). Urinalysis showed a markedly high urinary fluoride level in 14 of 23 patients (60.9%). Patients were followed up for an average duration of 4 years, 5 months. Paired t-test showed that the JOA score was slightly but nonsignificantly increased relative to preoperative measurement 3 days after surgery (P = 0.0829) and significantly increased at the end of follow-up (P = 0.0001). In conclusion, Fluorosis can cause ossification of thoracic ligamentum flavum, as well as other ligaments. Comparing with other OLF series, a larger number of spinal segments were involved. The diagnosis of skeletal fluorosis was made by the epidemic history, clinical symptom, imaging study findings, and urinalysis. En bloc laminectomy decompression was an effective method. 相似文献
995.
996.
目的 通过调查了解实习护生手卫生知识情况,分析手卫生依从性的影响因素.方法 采用自行设计的问卷对101名护生进行手卫生知识、行为进行调查.结果 手卫生知识掌握程度不够理想,手卫生态度行为处于较好水平,经过临床带教培训,护生操作后的态度和行为得分高于操作前(P<0.01),有创操作态度和行为得分高于无创操作和不接触患者的操作(P<0.01);护生自身学历和带教老师是影响其手卫生的重要因素.结论 实习护生的手卫生知识了解欠全面,应促进护生完善相关手卫生知识的补充与培训,手卫生态度和行为方面总体较好,但对于干燥双手和不同环境时的手卫生态度和行为欠佳,应加强临床带教中手卫生知识培训. 相似文献
997.
Zhu LX Liu J Ye Y Xie YH Kong YY Li GD Wang Y 《World journal of gastroenterology : WJG》2004,10(17):2488-2492
AIM: TO investigate the immunogenicity of candidate DNA vaccine against hepatitis C virus (HCV) delivered by two plasmids expressing HCV envelope protein 1 (El) and envelope protein 2 (E2) antigens respectively and to study the effect of CpG adjuvant on this candidate vaccine.METHODS: Recombinant plasmJds expressing HCV EI and E2 antigens respectively were used to simultaneously inoculate mice with or without CpG adjuvant. Antisera were then collected and tJters of antJ-HCV antibodies were analyzed by ELISA. One month after the last injection, animals were sacrificed to prepare single-cell suspension of splenocytes.These cells were subjected to HCV antigen specific proliferaion assays and cytokine secretion assays to evaluate the cellular immune responses of the vaccinated animals.RESULTS: Antibody responses to HCV EI and E2 antigens were detected in vaccinated animals. Animals receiving CpG adjuvant had slightly lower titers of anti-HCV antibodies in the sera, while the splenocytes from these animals showed higher HCV-antigen specific proliferation. Analysis of cytokine secretion from the splenocytes was consistent with the above results. While no antigen-specific IL-4 secretion was detected for all vaccinated animals, HCV antigen-specific INF-γ, secretion was detected for the splenocytes of vaccinated animals. CpG adjuvant enhanced the secretion of INF-γ, but did not change the profile of IL-4 secretion.CONCLUSION: Vaccination of mice with plasmids encoding HCV E1 and E2 antigens induces humoral and cellular immune responses. CpG adjuvant significantly enhances the cellular immune response. 相似文献
998.
Wang L Su W Liu Z Zhou M Chen S Chen Y Lu D Liu Y Fan Y Zheng Y Han Z Kong D Wu JC Xiang R Li Z 《Biomaterials》2012,33(20):5107-5114
Most hepatocellular carcinoma (HCC) therapies fail to target cancer stem cells (CSCs) and monitor cancer progression or regression. The purpose of this study was to evaluate the possibility of cancer imaging and simultaneously monitoring targeted therapy in a single animal by anti-CD44 antibody-mediated liposomal nanoparticle. In this study, an in situ liver tumor model was applied for therapy by injecting 1.0 × 10(6) HepG2 cells carrying a reporter system encoding a double fusion (DF) reporter gene consisting of firefly luciferase (Fluc) and green fluorescent protein (GFP) into the liver of NOD/SCID mice. A strategy was developed which specifically targeted HCC via anti-CD44 antibody-mediated liposomal nanoparticle delivery, loaded of either doxorubicin (Dox) or a triple fusion (TF) gene containing the herpes simplex virus truncated thymidine kinase (HSV-ttk) and renilla luciferase (Rluc) and red fluorescent protein (RFP). The NOD/SCID mice were subsequently treated with ganciclovir (GCV) and the growth status of tumor was monitored by optical bioluminescence imaging (BLI) of Fluc and specific targeting of the liposomal nanoparticle was tracked by Rluc imaging. CD44 antibody-mediated liposomal nanoparticle, loaded of TF plasmids, were shown to be useful for monitoring and evaluating targeting efficacy and gene therapy by non-invasive molecular imaging. Here, we demonstrate the time intensive preclinical steps involved in molecular target identification, validation, and characterization by dual molecular imaging. This targeted and traceable therapeutic strategy has potential advantages to overcome the problems of conventional tumor therapy and may open a new application for the treatment of HCC by targeting CSCs. 相似文献
999.
目的 分析普通外科老年患者手术后切口感染的影响因素,为医院感染的防治提供理论依据.方法 回顾性分析306例普通外科手术的老年患者的临床病历资料,分析手术后切口感染的危险因素.结果 306例老年患者手术后切口感染发生率为10.5%(32/306).210例合并慢性病患者手术后切口感染发生率显著高于96例非合并慢性病患者[12.9%(27/210)比5.2%(5/96)],92例夏季手术患者手术后切口感染发生率显著高于83例春季、71例秋季、60例冬季患者[16.3%(15/92)比8.4%(7/83)、8.5%(6/71)、6.7%(4/60)],120例手术时间>120 min患者手术后切口感染发生率显著高于186例手术时间≤120 min患者[15.0%(18/120)比7.5%(14/186)],162例急诊手术患者手术后切口感染发生率显著高于144例非急诊手术患者[12.3%(20/162)比8.3%(12/144)],差异均有统计学意义(P<0.05).结论 合并慢性病、夏季、手术时间> 120 min、急诊手术是普通外科老年患者手术后切口感染的危险因素. 相似文献
1000.
Shuai Zhang Yubo Liu Mingyang Ma Zheng Cao Xiangpeng Kong Wei Chai 《Orthopaedic Surgery》2022,14(8):1912
BackgroundRobotic‐assisted technology may be useful in hip revision cases with acetabular defects. However, data on the use of robotic‐assisted technology for such complex diseases is lacking.Case PresentationThis case study described the adoption of MAKO robotic‐assisted treatment of revision total hip arthroplasty (THA) combined with severe acetabular defect (Paprosky type IIIB). Robotic‐assisted technology accurately achieved preoperative planning; the acetabular component and augment were placed in the original position and angle as planned. Robotic‐assisted acetabular reaming was successful in a single pass, preserving the remaining acetabular bone mass very well with no procedure‐related complications. The Harris Hip Score (HHS) at 6 months postoperatively was 84 and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index was 24.ConclusionRobotic‐assisted technology can help in the accurate reconstruction of acetabular defect in complex hip revision surgery. 相似文献