全文获取类型
收费全文 | 33979篇 |
免费 | 3066篇 |
国内免费 | 2457篇 |
出版年
2024年 | 89篇 |
2023年 | 418篇 |
2022年 | 812篇 |
2021年 | 1326篇 |
2020年 | 963篇 |
2019年 | 1181篇 |
2018年 | 1158篇 |
2017年 | 797篇 |
2016年 | 1193篇 |
2015年 | 2021篇 |
2014年 | 2254篇 |
2013年 | 2494篇 |
2012年 | 3041篇 |
2011年 | 2837篇 |
2010年 | 1691篇 |
2009年 | 1505篇 |
2008年 | 1833篇 |
2007年 | 1646篇 |
2006年 | 1510篇 |
2005年 | 1240篇 |
2004年 | 1159篇 |
2003年 | 986篇 |
2002年 | 893篇 |
2001年 | 719篇 |
2000年 | 657篇 |
1999年 | 586篇 |
1998年 | 326篇 |
1997年 | 315篇 |
1996年 | 297篇 |
1995年 | 248篇 |
1994年 | 265篇 |
1993年 | 180篇 |
1992年 | 320篇 |
1991年 | 295篇 |
1990年 | 246篇 |
1989年 | 229篇 |
1988年 | 192篇 |
1987年 | 156篇 |
1986年 | 146篇 |
1985年 | 151篇 |
1984年 | 142篇 |
1983年 | 103篇 |
1982年 | 90篇 |
1980年 | 59篇 |
1979年 | 75篇 |
1978年 | 69篇 |
1977年 | 58篇 |
1976年 | 67篇 |
1975年 | 63篇 |
1974年 | 75篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
991.
为探讨细胞因子基因(人IL-2、IL-6)转导对于肿瘤细胞膜MHC抗原及细胞膜糖蛋白表达调控的影响,本文利用脂质体介导的方法,将含人IL-6、IL-2基因的逆转录病毒载体分别导入人乳腺癌细胞系MCF-7细胞中,采用间接免疫荧光染色流式细胞仪测定法,对基因转导的瘤细胞细胞膜糖蛋白及MHC抗原表达进行测定。结果表明经两种基因修饰的MCF-7细胞MHCⅠ型抗原表达均获得增强,此外,基因转导细胞可程度不同地表现出细胞膜多种糖蛋白表达的变化。提示肿瘤细胞膜抗原及糖蛋白表达的改变可能是细胞因子基因转导影响肿瘤细胞免疫原性的重要结构基础。 相似文献
992.
为大量制备β-NGF,构建了一种稳定、高效表达重组人神经生长因子(Recombinant human nerve growth factor,rh-β-NGF)的真核表达载体及含该重组载体的HEK293细胞株。首先,构建重组质粒p CMV-β-NGF-IRES-dhfr并转染至HEK293细胞系,用MTX加压筛选和有限稀释法进行选择,获得高效表达rh-β-NGF的单克隆重组细胞株;随后逐步降低血清培养,最终使细胞株完全适应无血清培养基并稳定表达rh-β-NGF;SDS-PAGE分析该表达产物,可见相对分子质量约13 k Da的条带,纯度大于50%,经质谱法测定得到其肽图谱与理论序列完全匹配,接着利用离子交换层析和分子筛层析纯化rh-β-NGF;最后进行重组细胞株表达效率和表达稳定性检测,表明重组细胞株可稳定、高效表达rh-β-NGF,其分泌效率大于20 pg/(cell?d),并能诱导PC12细胞的分化,具有良好的生物学活性。 相似文献
993.
994.
Jou SB Lin HM Pan H Chiu YL Li SY Lee CC Hsiao KM 《Proceedings of the National Science Council, Republic of China. Part B, Life sciences》2001,25(1):40-44
Myotonic dystrophy (DM) is an inherited, autosomal dominant muscular disease which is primarily caused by a CTG trinucleotide expansion mutation on chromosome 19q13.3. The size of this trinucleotide repeat is related both to the age of onset and to the severity of the clinical manifestation. This disease is very rare in Taiwan, and clinical and genetic study on DM has not yet been documented in this area. Here, we present both clinical features and degrees of CTG expansion for a Taiwanese DM family. All of the DM patients examined in this family showed obvious clinical manifestations by age 30, which included facial and limb muscle weakness with atrophy, myotonia, and ptosis. In addition, individual DM members also exhibited variable phenotypes, which may reflect the complexity of the pathogenic mechanism. Because the collection of blood specimens was considered to be an invasive procedure, a genetic study on this DM family was performed using buccal cells. Our results confirmed that four members showing classic symptoms of DM had CTG repeat expansion in the DMI locus, and that one member with ptosis and minor muscle weakness in the right foot was a normal homozygote for CTG repeat. These data demonstrate that buccal cells can provide clear and reliable results, and thus, are suitable for a family study of DM. 相似文献
995.
996.
Xiaohong Gu Yuechun Li Kaixin Chen Xingang Wang Zhongyu Wang Hao Lian Yuanzheng Lin Xing Rong Maoping Chu Jiafeng Lin Xiaoling Guo 《Journal of cellular and molecular medicine》2020,24(13):7515-7530
Human umbilical cord mesenchymal stem cell‐derived exosomes (hucMSC‐exosomes) have been implicated as a novel therapeutic approach for tissue injury repair and regeneration, but the effects of hucMSC‐exosomes on coxsackievirus B3 (CVB3)‐induced myocarditis remain unknown. The object of the present study is to investigate whether hucMSC‐exosomes have therapeutic effects on CVB3‐induced myocarditis (VMC). HucMSC‐exosomes were identified using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM) and Western blot. The purified hucMSC‐exosomes tagged with PKH26 were tail intravenously injected into VMC model mice in vivo and used to administrate CVB3‐infected human cardiomyocytes (HCMs) in vitro, respectively. The effects of hucMSC‐exosomes on myocardial pathology injury, proinflammatory cytokines and cardiac function were evaluated through haematoxylin and eosin (H&E) staining, quantitative polymerase chain reaction (qPCR) and Doppler echocardiography. The anti‐apoptosis role and potential mechanism of hucMSC‐exosomes were explored using TUNEL staining, flow cytometry, immunohistochemistry, Ad‐mRFP‐GFP‐LC3 transduction and Western blot. In vivo results showed that hucMSC‐exosomes (50 μg iv) significantly alleviated myocardium injury, shrank the production of proinflammatory cytokines and improved cardiac function. Moreover, in vitro data showed that hucMSC‐exosomes (50 μg/mL) inhibited the apoptosis of CVB3‐infected HCM through increasing pAMPK/AMPK ratio and up‐regulating autophagy proteins LC3II/I, BECLIN‐1 and anti‐apoptosis protein BCL‐2 as well as decreasing pmTOR/mTOR ratio, promoting the degradation of autophagy flux protein P62 and down‐regulating apoptosis protein BAX. In conclusion, hucMSC‐exosomes could alleviate CVB3‐induced myocarditis via activating AMPK/mTOR‐mediated autophagy flux pathway to attenuate cardiomyocyte apoptosis, which will be benefit for MSC‐exosome therapy of myocarditis in the future. 相似文献
997.
998.
一株新城疫病毒新强毒株(NL)的分离鉴定及F基因序列测定 总被引:3,自引:0,他引:3
1996年5月以来,1我国各地鸡群暴发鸡新城疫,其发病特征是常规疫苗免疫无效。为研究此病毒的特征,对从华东某地分离的新城疫病毒毒株(NL株)进行了生物学研究。利用RT-PCR技术扩增出NL株融合蛋白基因(F基因)全序列,并测序。结果表明,NL株平均致鸡胚死亡时间最短的,其尿囊液中病毒滴度可达2^9,表明病毒繁殖速度快。F基因测序结果表明,NL株为一强毒株,与现今国外已发表的NDV株F基因同源性在8 相似文献
999.
Lipopolysaccharide is strongly associated with septic shock, leading to multiple organ failure. It can activate monocytes and macrophages to release proinflammatory mediators such as tumor necrosis factor- (TNF-), interleukin-1 (IL-1), and nitric oxide (NO). The present experiments were designed to induce endotoxin shock by an intravenous injection ofKlebsiella pneumoniae lipopolysaccharide (LPS, 10 mg/kg) in conscious rats. Arterial pressure and heart rate (HR) were continuously monitored for 48 h after LPS administration. N-Acetyl-cysteine was used to study its effects on organ damage. Biochemical substances were measured to reflect organ functions. Biochemical factors included blood urea nitrogen (BUN), creatinine (Cre), lactic dehydrogenase (LDH), creatine phosphokinase (CPK), aspartate transferase (GOT), alanine transferase (GPT), TNF-, IL-1, methyl guanidine (MG), and nitrites/nitrates. LPS caused significant increases in blood BUN, Cre, LDH, CPK, GOT, GPT, TNF-, IL-1, MG levels, and HR, as well as a decrease in mean arterial pressure and an elevation of nitrites/nitrates. N-Acetylcysteine suppressed the release of TNF-, IL-1, and MG, but enhanced NO production. These actions ameliorate LPS-induced organ damage in conscious rats. The beneficial effects may suggest a potential chemopreventive effect of this compound in sepsis prevention and treatment. 相似文献
1000.