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Arancha C Ruíz-Llorente S Cascón A Osorio A Martínez-Delgado B Benítez J Robledo M 《Journal of human genetics》2002,47(4):190-195
Until now, the study of the multiple endocrine neoplasia type 1 (MEN1) gene in patients suspected of having the disease was expensive and laborious due to the large size of the gene. We have
optimized the conformation-sensitive gel electrophoresis (CSGE) technique to analyze by four rather simple multiplex PCR reactions,
and a single electrophoresis run, the entire coding region of the MEN1 gene, plus the exon–intron boundaries. This improvement of the CSGE technique was confirmed as an effective procedure for
screening for the MEN1 gene by detecting ten previously known MEN1 gene mutations and four polymorphisms. The MEN1 gene of 12 patients with unknown mutations was then screened, and an abnormal CSGE profile was identified in 10/12 cases.
Subsequent DNA sequencing demonstrated 3 of them to be novel mutations (E45K, 4479delACAG, 6073insC) and 7 to have been previously
reported; in the remaining 2 patients, we confirmed the absence of any alteration of the coding sequence of MEN1. Mutation screening of the MEN1 gene using CSGE was demonstrated to be a fast, simple, and inexpensive method to study patients suspected of having MEN1
disease.
Received: November 29, 2001 / Accepted: January 28, 2002 相似文献
3.
Rachael Goodman-Williams Rebecca Campbell Dhruv B. Sharma Steven J. Pierce Hannah Feeney Giannina Fehler-Cabral 《Journal of trauma & dissociation》2019,20(3):288-303
ABSTRACTHundreds of thousands of previously untested sexual assault kits (SAKs) have been uncovered in police property storage facilities across the United States, representing a national failure in institutional response to sexual assault. Faced with this discovery, jurisdictions must now decide if and how they should test these kits. Some stakeholders have suggested prioritizing kits for testing by victim, offender, or assault characteristics, based on the belief that these characteristics can predict the likely utility of DNA testing. However, little research has examined the empirical merits of such prioritization. To address this gap in the literature and inform SAK testing policies, we randomly sampled 900 previously untested SAKs from Detroit, MI. The sampled SAKs were submitted for DNA testing, and eligible DNA profiles were entered into Combined DNA Index System (CODIS), the federal DNA database. Police records associated with each SAK were coded for victim, offender, and assault characteristics, and logistic regression analyses were conducted to test whether these characteristics predict which SAKs yield DNA profiles that match (“hit”) to other criminal offenses in CODIS. Testing this sample of previously-untested SAKs produced a substantial number of CODIS hits, but few of the tested variables were significant predictors of CODIS hit rate. These findings suggest that testing all previously-unsubmitted kits may generate information that is useful to the criminal justice system, while also potentially addressing the institutional betrayal victims experienced when their kits were ignored. 相似文献
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A new method of STR interpretation using inferential logic -development of a criminal intelligence database 总被引:37,自引:0,他引:37
P. Gill A. Urquhart E. Millican N. Oldroyd S. Watson R. Sparkes C. P. Kimpton 《International journal of legal medicine》1996,109(1):14-22
Abstract A short tandem repeat (STR) system consisting of seven multiplexed loci has recently been introduced in the UK to support a National strategy to create large DNA databases for criminal intelligence purposes. The process uses automated sequencers, employing dye-labelled primers. Identification of tetrameric loci such as HUMTH01 are straightforward. Sizing windows are estimated by running a series of control allelic ladders on several gels and unknown samples are designated if they fall within a defined window. However, utilisation of complex STRs (eg. D21 S 11) characteristically have common variants which differ by just 2 bp. In addition, rare alleles are encountered which may differ by just 1 by from a common variant. To assist with the identification of alleles, we have introduced a series of allelic ladders, so that direct comparisons with unknown samples can be made on the same gel. To designate an allele, it should be within 0.5 by of an allelic ladder marker. Not all alleles (in particular rare alleles) can be included within an allelic ladder, however their expected positions can be easily calculated by reference to existing alleles in the ladder. Measurement of band shift is also a useful diagnostic tool. A series of guidelines are described to enable reliable allelic identification. These guidelines can be converted into computer programmes, which form the basis of an expert system. 相似文献
6.
NOVA16急诊生化分析仪试剂包中A、R、H液的研制与应用 总被引:1,自引:0,他引:1
目的 研制NOVA16全自动急诊生化分析仪试剂包中A,R,H液。方法 根据电极法测定血浆(或全血)中Na^+、K^+、Cl^-、TCO2、Glu、Urea、Cr等物质浓度(活度)和红细胞压积(Hematocrit,Hct)的原理及标准液配制的有关要求,配制类似血浆水的A液、CO2释放液H及参比液R。结果 自配A,R,H液与原装试剂的主要理化指标:pH、渗透压、电极斜率、线性范围及试剂的稳定性等方面 相似文献
7.
目的用相同标本比较国内外结核菌聚合酶链反应诊断试剂盒的临床检测效果,并初步探讨结核菌聚合酶链反应荧光诊断试剂盒在临床应用中定量检测结核菌数量的可行性。方法用聚合酶链反应技术,荧光探针杂交技术、酶联免疫反应技术以及常规细菌学方法检测并经双盲编号的84份临床痰标本和8份含痰结核菌标准品。结果在59例结核病患者痰标本中,国产PCR-酶联免疫检测试剂盒与PCR荧光试剂盒分别检出26例阳性和24例阳性,国外试剂盒检出24例阳性;在25例非结核病患者痰标本中,国产PCR-酶联免疫检测试剂盒与PCR荧光试剂盒分别检出21例真阴性和24例真阴性,国外试剂盒检出23例真阴性;细菌学各级别检查结果与PCR荧光检测CT值不呈递减关系。结论国内外结核菌聚合酶链反应诊断试剂盒对临床标本的检测结果无显著性差异;结核菌聚合酶链反应荧光诊断试剂盒尚不能定量检测痰标本中的结核菌数量,有待更多的临床数据加以证实。 相似文献
8.
微流芯片检测流感病毒多重逆转录聚合酶链反应产物的实验研究 总被引:1,自引:0,他引:1
[目的 ] 建立应用微流芯片检测甲 1型、甲 3型、乙型流感病毒多重逆转录聚合酶链反应 (mRT -PCR)产物的方法 ,在mRT -PCR扩增核酸的基础上自动化灵敏的定量检测扩增产物 ,快速检测甲亚型、乙型流感病毒 ,帮助临床快速诊断和鉴别诊断其他呼吸道病毒感染 ,以及明确流感病毒在人群中感染、流行情况。 [方法 ] 采用经MDCK细胞分离培养的甲 1型、甲 3型、乙型流感病毒毒株病毒液 ,使用 3组特异引物经mRT -PCR扩增核酸 ,扩增产物分别采用毛细管电泳技术经Caliper10 0 0微流芯片分析仪自动化检测和经 2 %琼脂糖凝胶电泳法检测。 [结果 ] 设计三组引物对相应甲 1型、甲 3型、乙型流感病毒靶基因的mRT -PCR扩增产物片段分别为 43 0bp、2 10bp、3 91bp ,本实验mRT -PCR扩增产物经 2 %琼脂糖凝胶电泳 ,与Marker比照 ,DNA条带基本在此位点附近 ;产物采用毛细管电泳法经Caliper10 0 0微流芯片分析仪后 ,分别于 413bp、2 0 3bp、3 79bp处出现陡峭的峰 ,如图所示。 [结论 ] 应用微流芯片采用毛细管电泳法可对流感病毒mRT -PCR产物进行定位及相对定量 ,有助于快速诊断流感病毒感染 ,有助于对流感的监测。 相似文献
9.
耳炎差异球菌与成人分泌性中耳炎的相关性研究 总被引:1,自引:0,他引:1
目的探讨耳炎差异球菌与成人分泌性中耳炎的关系。方法采用多重PCR的方法,检测39例(42耳)成年分泌性中耳炎患者的中耳积液中耳炎差异球菌以及三种常见细菌[肺炎链球菌(S.pneumoniae)、流感嗜血杆菌(H.influenzae)和卡他莫拉杆菌(M.catarrhalis)]的DNA,就细菌DNA存在与成人分泌性中耳炎患者上呼吸道感染(简称上感)史、病程、积液性质和抗生素服用史之间的关系进行分析。结果收集到的42份中耳积液标本中有5份(11.9%)检测到有耳炎差异球菌DNA存在。急性分泌性中耳炎者有1例(3.6%)测得耳炎差异球菌DNA存在,慢性者则有4例(28.6%),两者差异有显著性意义。浆液性中耳积液有3例(13.0%)测得耳炎差异球菌DNA存在,粘液性中耳积液则有2例(10.5%)为阳性,两者差异有显著性意义。抗生素使用史对耳炎差异球菌DNA检出率的影响无统计学意义。结论细菌参与部分成人分泌性中耳炎的发病,耳炎差异球菌可能是其致病菌之一;且慢性分泌性中耳炎中耳积液的持续存在可能与耳炎差异球菌感染有关。 相似文献
10.
目的 建立改良分子信标,多重实时PCR同时检测沙门菌和志贺菌的快速方法,应用于食源性致病菌的快速诊断。方法 根据GenBank公布的沙门菌侵袭性基因invA和ssaR基因,分别设计一对引物和改良分子信标探针,用同色荧光标记,用于同体系检测沙门菌。志贺菌根据ipaH基因的保守序列,设计引物和改良分子信标探针,加入沙门菌检测体系中,建立三重实时PCR一改良分子信标检测体系,应用于同时对沙门菌、志贺菌食物中毒的快速诊断和门诊肠道致病菌的检测。结果 改良分子信标一多重实时PCR反应体系DNA灵敏度为69~93fg/μl。菌液灵敏度为32~64CFU/ml或1~2CFu/PCR反应体系,无交叉反应。该反应体系同时检测134株沙门菌和67株志贺菌,均出现特异的荧光信号,两种细菌检测互不干扰。对细菌性食物中毒样本等共1100份同时进行沙门菌和志贺菌检测,569份沙门菌实时PCR阳性,其中551份沙门菌培养阳性;42份志贺菌实时PCR阳性,其中41份志贺菌培养阳性。从样品处理到检测结果仅需时间2h至1d。结论 改良分子信标-多重实时PCR检测体系快速、灵敏度高,特异性强,可用于沙门菌和志贺菌食物中毒的快速诊断,伤寒、痢疾等肠道传染病的初筛及预防医学门诊的健康人群体检,为食源性疾病的分子流行病学调查提供新的检测手段。 相似文献