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991.
992.
本研究构建人细胞间黏附分子1(ICAM-1)全长基因的真核表达载体pEGFP—CI—ICAM—1,转染中国仓鼠卵巢细胞(CHO—K1)细胞株,并检测其在cH0细胞中的表达及与Molt-4细胞的结合。采用RT—PCR法从健康人外周血中分离单个核细胞,钓取ICAM-1全长基因(1622bp),与pMD18-T载体连接做全自动序列测定。将测序正确的克隆质粒pMD18-T—ICAM-1和表达载体pEGFP—C1分别用HindⅢ和Ⅰ进行双酶切,应用基因重组技术构建ICAM-1全长基因真核表达载体pEGFP—C1-ICAM-1,质粒经Hindm和SacⅡ双酶切和PCR电泳鉴定后,采用脂质体转染法转染CHO细胞,并进行G418筛选。用RT—PCR、流式细胞术和荧光显微术检测ICAM-1-GFP的表达及亚细胞的定位,用检测ICAM-1-GFP/CHO细胞与Molt-4细胞的结合能力评价ICAM.1-GFP融合蛋白的功能。结果表明:重组质粒经限制性酶切鉴定得到与ICAM—1全长基因长度-致(1622bp)的酶切产物;测序分析证实,PCR产物与GenBank上登录的ICAM-1基因(NM-000201)序列完全-致,表明成功地完成了ICAM-1的扩增和表达载体的构建;荧光显微镜下可见转染的CHO细胞有绿色荧光蛋白的表达,表达的融合蛋白较均匀地分布于整个细胞:FACS检测ICAM.1-GFP的荧光转染率为(13±5.5)%,表明ICAM—1—GFP基因进入到CHO细胞并获得了有效表达,成功构建了ICAM-1-GFP/CHO细胞,并且ICAM-1-GFP/CHO细胞能够结合PMA处理的Molt-4细胞。结论:成功构建了ICAM-1-GFP真核表达载体,构建的ICAM—1—GFP真核表达载体在CHO细胞内稳定表达,/CAM—1—GFP/CHO细胞能与M0lt-4细胞结合,这为进-步研究ICAM-1分子的功能打下基础。 相似文献
993.
双色双融合荧光原位杂交探针检测慢性髓系白血病Bcr/Ab1基因重排的研究 总被引:1,自引:0,他引:1
本研究探讨双色双融合荧光原位杂交技术(DC-DF-FISH)在慢性髓系白血病(CML)中的应用价值,应用常规R-显带技术、DC-DF-FISH、RT—PCR技术检测41例CML患者的染色体核型及bcr/abl融合基因。结果表明,对于初诊CML的18例患者,R-显带显示Ph染色体阳性检出率为94.4%(17/18),DC—DF—FISH阳性检出率也为94.4%(17/18),对于治疗后CML的18例患者,R-显带显示14例有可分析分裂相,其中有ll例存在Ph染色体,阳性检出率为78.6%(11/14);而用DC—DF—FISH检测治疗后患者的阳性率为94.4%(17/18);移植后的5例患者R-显带均未检出Ph染色体,而FISH检测出1例bcr/abl基因阳性,RT-PCR证实了FISH的检测结论,但在移植患者中,RT-PCR无阳性发现。结论:双色双融合荧光原位杂交技术具有高度的准确性、可靠性,是检测CML患者bcr/abl基因重排的可靠方法,适用于CML的诊断、疗效判定及微小残留病灶的检测。 相似文献
994.
目的 探索西藏农牧区居民结核病防治知识现状及其影响因素,为西藏农牧区开展有针对性的结核病健康教育提供依据。方法 采用分层整群抽样法,抽取西藏4个市中8个村落的1 466名农牧区居民进行入户面对面问卷调查。采用率统计描述,最优尺度回归统计分析。结果 西藏农牧区居民的结核病防治知识总知晓率为34.9%。知识得分的影响因素按重要性依次为,居住地、是否接受过结核病防治健康教育、文化程度、年龄。结论 西藏农牧区居民的结核病防治知识水平低,应均衡推进各市农牧区健康教育工作,以青年人、文化程度较低者为重点人群。 相似文献
995.
Stem cell and regenerative science applications in the development of bioengineering of renal tissue
A rising number of patients with acute and chronic renal failure worldwide have created urgency for clinicians and investigators to search out alternative therapies other than chronic renal dialysis and/or organ transplantation. This review focuses on the recent achievements in this area, and discusses the various approaches in the development of bioengineering of renal tissue including recent discoveries in the field of regenerative medicine research and stem cells. A variety of stem cells, ranging from embryonic, bone marrow, endogenous, and amniotic fluid, have been investigated and may prove useful as novel alternatives for organ regeneration both in vitro and in vivo. Tissue engineering, developmental biology, and therapeutic cloning techniques have significantly contributed to our understanding of some of the molecular mechanisms involved in renal regeneration and have demonstrated that renal tissue can be generated de novo with similar physiologic functions as native tissue. Ultimately all of these emerging technologies may provide viable therapeutic options for regenerative medicine applications focused on the bioengineering of renal tissue for the future. 相似文献
996.
Hall R Wartman D Jellinek N Robinson-Bostom L Telang G 《Journal of cutaneous pathology》2008,35(Z1):14-16
Lichen planus (LP) is an inflammatory dermatitis of idiopathic origin that can involve the skin, mucous membranes, hair and nails. Histologically, LP is characterized by compact orthokeratosis, wedge-shaped hypergranulosis, irregular acanthosis, damage to the basal cell layer and a band-like inflammatory infiltrate in the upper dermis. Lymphocytes are the predominant cells making up the infiltrate, along with a few macrophages, eosinophils and plasma cells. In addition, melanophages are often found in the upper dermis adjacent to the damaged basal cells.(1) We describe a patient with a lesion of the toenail clinically and histopathologically consistent with LP, but with a band-like inflammatory infiltrate composed primarily of plasma cells. Previously, only three other cases of LP with plasma cell predominant infiltrate have been reported, none of which involved the nail matrix. 相似文献
997.
Jellinek M 《Medical hypotheses》2008,71(4):515-526
Cited research papers support the main hypothesis that selected publications supply sufficient information for a combined multi-level treatment strategy against cancer that will also strengthen the host. The three major elements of the proposal are: (A) metastasis being separate from tumor growth requires specific antimetastatic treatments. For this, manipulation of the composition of phospholipids will alter cellular charge characteristics which are instrumental in adhesion. (B) Formate metabolism is at the center of many activities that are controlling tumor growth. The rational and consequences of this are as follows. Supply of formate depends mainly on serine, and consumption on conversion to CO(2) yielding needed NADPH. The remainder is used to complete IMP configuration with 5-aminoimidazole-4-carboxamide ribonucleotide (ZMP). At homeostasis residual ZMP activates AMP-activated protein kinase (AMPK) to curb growth promoting phosphatidylinositol-3-kinase (PI3PK). Residual ZMP also activates the oxidation of choline to betaine supplying methyl groups needed for global methylation of DNA while increased oxidation of choline also alters cellular phospholipid composition (refer to metastasis). At low formate level, increased accumulated ZMP becomes pyrophosporylated to ZTP. AMPK activation shifts to PI3PK activity for insulin action restoring formate supplied by serine derived from glycolysis. Increased NADPH-generating glucose-6-phosphate dehydrogenase is diminishing NADP(+) required for dehydrogenation of formate. This is restoring the formate balance while lowering ZMP levels to that of homeostasis. Evidence suggests that transformed cells exceed up-regulation of formate thus suppressing all ZMP accumulations resulting in limited AMPK activation, cessation of choline oxidation to betaine and loss of global methylation of DNA. This scenario appears to be tied to tumor survival, a state that could be altered by metabolic interventions using mild agents as described in the research reports cited. (C) Because of a preponderance of pyrimidines in cancer supporting UTP requiring immune evasion, exogenous IMP may offset this imbalance and thus hinder tumor anti-immune activities while strengthen host immune functions. For studies to confirm the proposal, the overall expected result is that a combined administration of all these agents cited here will outperform any single agent considered so far for anticancer treatment. 相似文献
998.
DNA/子弹.基因枪转染的效率为10.5%±2.4%(n=3),电转染方法 为45.2%±5.6%(n=3),而脂质体方法 只有个别DC表达荧光蛋白,三者比较差异有统计学意义(P<0.01).基因枪方法 转染的DC能够保持其特殊细胞形态,细胞表而标志在转染中无明显变化,而电转染后细胞大量死亡.应用基因枪转染方法,基因疫苗质粒IgHV1-GM-CSF/pcDNA3.0在IX;中得到瞬时表达.结论 基因枪颗粒轰击可使基因疫苗在DC中成功表达,是临床细胞免疫治疗中DC转染的一种可行选择. 相似文献
999.
目的:探讨心梗后(PMI)患者递增负荷运动中生理反应与心电图ST段水平变化的关系,旨在根据服用和不服用β-受体阻滞剂患者的不同反应,为心脏康复中适宜运动强度的制定提供依据。方法:46名服用β-受体阻滞剂的和55名不服用β-受体阻滞剂的男性PMI患者,依据改良布鲁斯方案在活动跑台上进行递增负荷运动试验。运动中每30s测量一次摄氧量(VO2)和通气量(VE),每3min记录一次心率血压乘积(RPP)和血乳酸浓度(BL),并连续监测12导心电图。结果:(1)ST段下降水平与VO2、VE和BL的关系,两组患者间无显著差异(P>0.05)。但两组在相同RPP时,其ST段水平却存在显著差异(P<0.01);(2)ST段下降1mm时,患者的VO2、VE和BL分别约为30ml.kg-1.min-1、63L.min-1和5.3mmol.L-1;(3)ST段下降1mm时,服用和不服用β-受体阻滞剂患者的RPP分别为190beats.mmHg.100-1和230 beats.mmHg.100-1。结论:为了减少心肌缺血的发生,结果(2)和(3)中生理指标的数值,应被作为心脏康复中PMI患者运动强度的上限。 相似文献
1000.
Objective: Endodontic originated chronic apical periodontitis (AP) is an inflammatory disease of periapical tissue. High-sensitivity C-reactive protein (hsCRP) as an inflammatory marker and hemogram indexes provide valuable information to clinicians for diagnosis, screening and follow-up of various diseases. The aim of this study was to investigate AP in terms of its association with hemogram indices and hsCRP levels.Material and methods: Study includes 104 patients with AP and 40 participants as the control group. 160 teeth were diagnosed as AP through digital radiographic images and scored with respect to Periapical Index (PAI) scoring. Afterwards, patients were categorized into 3 grades in accordance with both the number and the severity of AP. AP grade 0 was considered for the control group with regard to a new scoring system. Patients with only one tooth involved with AP with a PAI score of 3 or 4 were categorized as an AP Grade 1, when a patient had more than one tooth with a PAI score of 3 or 4 he was classified as an AP Grade 2 and a patient with at least one tooth scored as a PAI 5 was rated as an AP Grade 3. Hemograms and hsCRP levels were measured for each individual to establish a correlation with inflammatory markers.Results: The neutrophil/lymphocyte ratio (NLR) levels of patients with AP Grade 3 were significantly higher than all other AP grades (p?<?.05). hsCRP levels in patients with an AP Grade 2 and 3 were higher than both AP Grade 0 and 1 (p?<?.05).Conclusions: hsCRP levels of patients were reliable predictive indicators for AP severity in correlation with the new proposed scoring system for AP. 相似文献