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Objective To detect the expression of type Ⅰ interferon in monocyte-derived dendritic cells(MoDCs)after Toll like receptor(TLR)3 triggered in patients with chronic hepatitis B(CHB),and to evaluate immune responses of CHB patients and its roles in the mechanisms of persistent infection of hepatitis B virus(HBV)and chronicity of hepatitis.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and purified using magnetic beads(plasma was saved simultaneously)from 26 CHB patients and 18 healthy volunteers(HV).Dendritic cells(DCs)were induced and proliferated in a culture medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin(rhIL-4).EX3s were stimulated with Poly Ⅰ:C and the supernatants were collected at 0 h and 24 h after stimulation.Type Ⅰ interferon(IFN-α and IFN-β)in plasma and supernatants were examined by enzyme linked immunosorbent assay (ELISA).Results The levels of type Ⅰ interferon in plasma were not significantly different in groups of HV and CH B.IFN-α and IFN-β expressions in supernatants before Poly Ⅰ:C stimulation were(80.00±16.15)ng/L,(36.39±13.90)ng/L in CHB group and(76.76±15.90)ng/L,(37.14±13.68)ng/L in HV group,respectively.And there were no statistical differences between two groups(t=1.651,t=0.178;both P>0.05).IFN-α expressions in supernatants at 24 h after stimulation in two groups were both higher than those before stimulation(at 0 h),but there were no statistical differences(t=1.534,t=1.243;both P>0.05).IFN-β expressions in supernatants at 24 h after stimulation in HV group was(54.57±16.80)ng/L,which was significantly higher than that at 0 h(37.14±13.68)ng/L(t=4.061,P<0.05).However,there was no significant difference at 24 h than tht at 0 h in CHB group(t=1.796,P>0.05).At 24 h after stimulation.IFN-β level was(54.57±16.80)ng/L in HV group,which was significantly higher than that[(41.64±12.57)ng/L]in CHB group(t=2.921,P<0.05).Conclusions Functions of MoDCs from CHB patients are impaired and MoDCs could not express type Ⅰ interferon normally.Expression of type Ⅰ interferon after TLR3 triggered in CHB patients is mainly IFN-β. 相似文献
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汉坦病毒汉城型浙37株G1、G2包膜糖蛋白基因重组体的构建及在真核细胞中的表达 总被引:2,自引:1,他引:2
目的:构建汉坦病毒浙37(Z37)株包膜糖蛋白基因G1、G2真核表达质粒,并在真核细胞中表达。方法:根据Z37M基因序列设计6条引物,分别以质粒pGEMZ37,pCUMZ37为模板,通过聚合酶链反应(PCR)获得G1及G2片段。将G1、G2片段经BamHⅠ、XhoⅠ双酶切片插入至真核表达载体pcDNA3.1( ),经酶切鉴定,并测序证实。以磷酸钙沉淀法分别将重组质粒转染COS-7细胞,用间接免疫荧光法(IFA)检测瞬时表达的蛋白。结果:获得分别含有编码汉坦病毒(HV)Z37株包膜糖蛋白G1、G2基因的重组质粒pcDNA3.1-g1,pcDNA3.1-G2;在转染的COS-7细胞内,用IFA可检测到细胞内有特异性荧光。结论:成功地构建了HV Z37株包膜糖蛋白G1、G2基因真核表达载体,并可在COS-7细胞中瞬时表达。 相似文献
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Objective To detect the expression of type Ⅰ interferon in monocyte-derived dendritic cells(MoDCs)after Toll like receptor(TLR)3 triggered in patients with chronic hepatitis B(CHB),and to evaluate immune responses of CHB patients and its roles in the mechanisms of persistent infection of hepatitis B virus(HBV)and chronicity of hepatitis.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and purified using magnetic beads(plasma was saved simultaneously)from 26 CHB patients and 18 healthy volunteers(HV).Dendritic cells(DCs)were induced and proliferated in a culture medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin(rhIL-4).EX3s were stimulated with Poly Ⅰ:C and the supernatants were collected at 0 h and 24 h after stimulation.Type Ⅰ interferon(IFN-α and IFN-β)in plasma and supernatants were examined by enzyme linked immunosorbent assay (ELISA).Results The levels of type Ⅰ interferon in plasma were not significantly different in groups of HV and CH B.IFN-α and IFN-β expressions in supernatants before Poly Ⅰ:C stimulation were(80.00±16.15)ng/L,(36.39±13.90)ng/L in CHB group and(76.76±15.90)ng/L,(37.14±13.68)ng/L in HV group,respectively.And there were no statistical differences between two groups(t=1.651,t=0.178;both P>0.05).IFN-α expressions in supernatants at 24 h after stimulation in two groups were both higher than those before stimulation(at 0 h),but there were no statistical differences(t=1.534,t=1.243;both P>0.05).IFN-β expressions in supernatants at 24 h after stimulation in HV group was(54.57±16.80)ng/L,which was significantly higher than that at 0 h(37.14±13.68)ng/L(t=4.061,P<0.05).However,there was no significant difference at 24 h than tht at 0 h in CHB group(t=1.796,P>0.05).At 24 h after stimulation.IFN-β level was(54.57±16.80)ng/L in HV group,which was significantly higher than that[(41.64±12.57)ng/L]in CHB group(t=2.921,P<0.05).Conclusions Functions of MoDCs from CHB patients are impaired and MoDCs could not express type Ⅰ interferon normally.Expression of type Ⅰ interferon after TLR3 triggered in CHB patients is mainly IFN-β. 相似文献
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近年来,人们运用诊断技术,特别是分子生物学技术诊断肝脏疾病。现就一些新的核酸检测技术、肝活检技术和血清标记物的临床应用做一总结。 相似文献
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患者,男,33岁。因反复高热2月余入院。患者2个月来反复出现发热,体温最高达40℃,在外院查血白细胞2.7×10~9/L,中性粒细胞0.68,先后给予“头孢哌酮、左氧氟沙星、青霉素、头孢曲松”等多种抗菌药物治疗,用药后体温能降至正常,但停药后再次发热,如此反复多次。1周前患者再次出现高热,伴咳嗽,咳黄脓痰,自服抗生素药效不佳。入院体检:体温38.3℃,血压110/70 mm Hg(1mm Hg= 0.133 kPa),皮肤、黏膜未见皮疹、出血点,全身浅表淋巴结未及,咽无充血,扁桃体无充血肿大。颈软,两肺呼吸音粗, 相似文献