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1.
由一株抗人C1-抑物单克隆抗体鼠杂交瘤细胞提取总RNA,合成2对与免疫蛋白可变区FR1和FR4互补的通用引物,经RT-PCR扩增出抗体的重链可变区和轻链可变区基因。 相似文献
2.
目的:克隆抗人CD154抗体轻重链可变区基因,并分析其核苷酸序列,为基因工程抗体的构建奠定基础。方法:从分泌能抑制免疫反应的抗人CD154单克隆抗体杂交瘤细胞株 7E8中提取总RNA,合成cDNA第一链后,经PCR扩增获得抗人CD154单抗轻链可变区 (VL)和重链可变区 (VH)基因,分别克隆入pUC18载体,并进行序列分析。结果:①抗体的轻链可变区基因全长为 341bp,编码113个氨基酸,归属于Ig的Vκ2基因,氨基酸序列分析结果显示轻链可变区含有明确的 4个骨架区和 3个抗原决定簇互补区,在第 2 3位和第 93位氨基酸为半胱氨酸,是与抗体二硫键形成有关的两个特征性氨基酸;②抗体的重链可变区基因全长为 354bp,编码118个氨基酸,归属于小鼠IgVH基因,D、J区基因分别属于DSP2.9和JH2,氨基酸序列分析结果显示,重链可变区含有明确的 4个骨架区和 3个抗原决定簇互补区,在第 2 3和第 97位氨基酸为半胱氨酸,是与抗体二硫键形成有关的两个特征性氨基酸。结论:经核苷酸序列分析证明所克隆的基因分别为抗体的轻、重链可变区基因. 相似文献
3.
Kazuya Takeda Shuhei Sakakibara Kazuo Yamashita Daisuke Motooka Shota Nakamura Marwa Ali El Hussien Jun Katayama Yohei Maeda Masanobu Nakata Shigeyuki Hamada Daron M. Standley Masaki Hayama Takashi Shikina Hidenori Inohara Hitoshi Kikutani 《The Journal of allergy and clinical immunology》2019,143(3):1163-1175.e15
4.
[摘要] 目的:研究Iα1 hs1,2 VNTR多态性与我国IgA肾病的相关关系。 方法: 采集419例肾活检证实的IgA肾病患者及其一级亲属、条件相当的201例健康志愿者血样,提取基因组DNA。用PCR产物直接电泳法鉴定Iα1 hs1,2 VNTR基因型,采用以家庭为基础的传递/不平衡分析(TDT)和单倍型相对危险度(HRR),以及病例-对照研究分析Iα1 hs1,2 VNTR多态性与我国IgA肾病的相关关系。 结果: ① TDT分析结果显示Iα1 hs1,2 VNTR B等位基因从杂合子父母向患者传递的频率显著高于预期值(101 Trios, χ2=6.818, P<0.01,扩展TDT分析也得到相同结果(164家庭, χ2=7.583, P<0.01)。②与TDT结果一致,HRR分析同样显示Iα1 hs1,2 VNTR B等位基因的过度传递 (P<0.05, χ2=4.122, HRR=1.180), 而BB基因型具有更强的患病倾向 (P<0.05, χ2=4.411, OR=1.538)。③病例-对照研究显示IgA肾病组B 等位基因频率显著高于正常对照组(χ2=6.968, P<0.05)。 结论: Iα1 hs1,2 VNTR基因多态性与我国IgA肾病患者的易感性相关。 相似文献
5.
目的评价重症医学科DRG分组的效能,发现存在的问题,为DRG付费或绩效考核提供建议。方法采用DRG分组器获得样本医院2016―2019年出院患者的分组结果。在2555名重症医学科出院患者中选取病例数排名前5位的DRG组作为DRG研究组。在351148名全院出院患者中筛选出DRG研究组的所有病例。对入组同一个DRG组的患者,根据其所属的出院科室分为全院组(包括重症医学科和非重症医学科出院患者)和重症医学科组(仅包括重症医学科出院患者)。通过计算住院费用的变异系数评价全院组和重症医学科组的组内同质性。结果在同一个DRG组中,重症医学科出院患者组住院费用的变异系数低于全院出院患者组,即对于同一个DRG分组病种,重症医学科出院患者单独评价时组内住院费用的同质性更高。结论应用DRG付费或绩效考核时,针对重症医学科出院的患者应考虑细化DRG分组或调整分组权重,以便形成更加科学合理的疾病分组。 相似文献
6.
A repeated measures procedure, variable intertrial interval (ITI) spatial alternation, was used to assess scopolamine effects on memory, and to compare effects of the drug on discrimination processes with effects on storage. Rats learned in two stages to press left and right levers in alternation on discrete trials separated by 5 different ITI's ranging from 2.5 to 40 s and presented in random order during the experimental session. In the first stage, alternating discrimination, alternation was controlled by a light on over the correct lever at the time of the trial; in the second stage, variable ITI spatial alternation, a centrally located panel light signalled all trials and alternation was controlled by stimuli from prior trials (memory). Alternation response occurrence declined moderately (but significantly) with increasing ITI duration in both the alternating discrimination and variable ITI spatial alternation stages; response occurrence was also significantly decreased by scopolamine treatment in both stages. Accuracy of alternating discrimination performance was not significantly altered by either ITI duration or scopolamine treatment. Accuracy of variable ITI spatial alternation performance on a trial varied inversely with the duration of the ITI that preceded the trial. Scopolamine treatment significantly reduced accuracy of lever pressing in variable ITI spatial alternation but did not alter the slope of the curves relating accuracy to ITI duration. These effects indicate that the drug impaired discrimination processes but did not alter memory storage. 相似文献
7.
目的 应用数目可变串联重复序列(VNTR)分子分型技术,对北京地区113株肺结核临床分离菌株进行分型研究,探讨北京地区菌株DNA多态性和基因型特征。方法 采用PCR和琼脂糖凝胶电泳技术对结核分支杆菌13个VNTR位点进行检测,并应用Gel-Pro analyzer 3.1软件和BioNumerics3.0软件进行结果分析。结果 113株结核分支杆菌可分为4个基因型(分别为Ⅰ、Ⅱ、Ⅳ和Ⅴ型),其中Ⅰ型占92.0%(104/113),其他3型所占比例很小,分别为Ⅱ型占4.4%(5/113),Ⅳ和Ⅴ型均为1.8%(2/113),而标准菌株H37Rv在分型中为独立的一个基因型,即Ⅲ型。结论 北京地区的结核分支杆菌存在基因多态性,其主要流行型为Ⅰ型。 相似文献
8.
《International journal of audiology》2013,52(3):259-267
The frequency response and loudness reduction characteristics of earmolds with variable venting valves (VVV) were investigated. Both side-branch and laterally vented earmolds were employed. Sound pressures were measured at 44 frequencies from 100 to 4 000 Hz in a modified HA-2 coupler with the VVV in four stages of opening: closed; ? open; ? open and 3/3 openThe effect of venting is primarily in the low frequencies. Little or no reduction in intensity is observed in the ‘speech frequency’ range and a modest amount is noted in the higher frequencies. The side-branch vented earmolds were more effective than the laterally vented earmolds. Calculated loudness reductions in phons were small. The effectiveness of the VVV, whether assessed by the frequency response or loudness reduction characteristics, is achieved within the first ? of opening: further opening has little effect. The utility of the VVV, especially to the geriatric hearing aid user, is questionable 相似文献
9.
10.
The controversy surrounding the findings that copy number variation, of the CCL3 encoding genes, influences HIV-1 infection and disease progression has been in part attributed to the variable results obtained from methods used for copy number evaluation. Like CCL3, the genes encoding the CC chemokine CCL4, also a natural ligand of the CCR5 receptor, are found to occur in population-specific multiple copy number and have been shown to play a protective role against HIV-1. This study evaluated the standard method of quantitative Real-Time PCR (qPCR) and droplet digital PCR (ddPCR) for CCL4L gene copy number determination. The CCL4 encoding genes are CCL4, occurring in two copies per diploid genome (pdg), and the non-allelic CCL4L genes, comprised of CCL4L1 and CCL4L2, which are both found in multiple copies pdg. Copy number of CCL4L, CCL4L1 and CCL4L2 was determined in a cohort of HIV-1-uninfected individuals from the South African Black (n = 23) and Caucasian (n = 32) population groups using qPCR and ddPCR. A stronger correlation between the number of CCL4L copies and the sum of CCL4L1 and CCL4L2 copies generated by ddPCR (r = 0.99, p < 0.0001) compared to qPCR (r = 0.87, p < 0.0001) was observed. Real-Time qPCR exhibited greater inaccuracy at higher copy numbers which is particularly relevant to our cohort of Black individuals who have a higher range of CCL4L copies (3–6) compared to Caucasians (0–4) and a higher population median (4 and 2, respectively). Medians and ranges of CCL4L1 (Black: 2, 0–4, Caucasian: 0, 0–2) and CCL4L2 (Black: 2, 1–5, Caucasian: 2, 0–3) were also higher in the Black population. Droplet digital PCR was shown to be a far superior method to qPCR for assessment of CCL4 gene copy number variation, the accuracy of which is essential for studies of the contribution of variable gene copy number to phenotypic outcomes of host infection and disease course. 相似文献