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21.
Objective To establish and characterize a novel human myeloid leukemia cell line SH-2. Methods Bone marrow mononuclear cells(BMMNC) isolated from a AML-M2 patient, who failed to ob-tain complete remission after chemotherapy and allogenic bone marrow transplantation were passed in a long term IMDM culture medium supplemented with 20% fetal calf serum. Stromal cells were retained and rh-IL-3was added in the culture system. A new human myeloid leukemia cell line SH-2 was successfully established with a cytogeuetic characteristics of a loss of Y chromosome(- Y), a derivative chromosome 16 resulting from unbalanced translocation between chromosome 16 and 17, monosome 17, trisomy 19 and p53 alteration. Vari-ous methods were employed to characterize SH-2 cell line. Results SH-2 cells has been maintained without cytokine and stromal cells for more than 3 years without EB virus and mycoplasma contamination. SH-2 cells had the basically same morphological, immunophenotypic and cytogenetic features as the patient' s leukemia cells did, such as myeloid morphology, an immunophenotype of CD13+ , CD33+ , CD56+ , CD16/56+ and a hypodiploid karyotype of 45, X, - Y, der(16) t(16;17) (q24;q12) , - 17, + 19, which were gradually de-creased and replaced by the near-tetraploid cells with a karyotype of 73 - 102 (80), XX, - Y, - Y, del (lq31) ×2, der(16)t(16;17) (q24;q12) ×2, - 17, - 17, + 19, + 19. FISH and multiple FISH delineated all the abnormalities and revealed a loss of one p53 allele due to monosomy 17. DNA direct sequencing detec-ted a point mutation of CAG to CAT at codon 576 of exon 5 in another p53 allele. RT-PCR showed that SH-2 cells expressed apoptosis-related genes (bcl-2, Fas, GST- π and p21) rather than MDR-related genes. Short tandem repeat PCR provided powerful evidence for the derivation of SH-2 cell line from the patient' s leukemia cells. SH-2 cells had certain colony formation and tumorigenic capacities in nude and SCID mice. Conclu-sion SH-2 is a new myeloid leukemia cell line with a unique biology background, and will provide a useful tool for leukemia research. 相似文献
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多色荧光原位杂交技术检测急性淋巴细胞白血病复杂核型异常 总被引:12,自引:0,他引:12
本研究建立多色荧光原位杂交(M—FISH)技术平台,探讨其在检测急性淋巴细胞白血病(ALL)复杂核型异常中的应用。联合应用常规细胞遗传学方法和M—FISH技术分析了5例伴有复杂核型异常的ALL患者。结果表明:M—FISH证实了原有的异常t(9;22)、t(1;19)和t(y;1),同时还发现了新的异常der(1)(1::3::7)、der(6)t(6;9)(q?;p13)、der(1)t(1;11)、der(12)t(1;12)、der(3)t(3;5)、der(2)t(2;16)、der(9)(9::18::7)和der(7)(9::18::7),并且纠正了原有的错误分析,其中der(9)(9::18::7)及der(7)(9::18::7)为世界上首例报道。结论:M—FISH在检测ALL复杂核型中的应用前景广阔,是进行精确染色体核型分析所不可缺少的先进手段。 相似文献
24.
i(7q)是恶性血液病中一种少见的非随机核型异常,主要见于急性淋巴细胞白血病(acute lymphoblastic leukemja,ALL),其次也可见于急性髓细胞白血病(acute myeloid leukernia,AML)、慢性髓细胞白血病(chronic myeloid leukernja,CML)和淋巴瘤。我们对17例伴有i(7q)异常恶性血液病患者的临床和细胞遗传学资料进行分析,以探讨其临床和预后特点。 相似文献
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目的 报告1例伴有低亚二倍体复杂异常的多发性骨髓瘤病例并探讨其临床和实验室特点.方法 采用骨髓细胞短期培养法制备染色体,用R显带技术进行核型分析.用13q14、p53、Rbl、lq21一系列单色探针和IgH/CCND1双色双融合探针对其进行荧光原位杂交检测.用流式细胞仪检测DNA含量.结果 R显带核型分析提示该患者5个细胞为包含35条染色体的低亚二倍体核型,3个细胞为低亚二倍体克隆的复制,另外4个细胞为正常核型.间期荧光原位杂交证实核型中存在1号、13号、14号、17号染色体单体,且显示marl来源于11号染色体并造成CCND1基因的扩增.流式细胞仪检测DNA含量显示其有低亚二倍体克隆峰,DNA指数为0.8426.结论 低亚二倍体核型在多发性骨髓瘤中发生率极低,荧光原位杂交技术是检测多发性骨髓瘤分子异常的可靠手段. 相似文献
27.
嗜酸性粒细胞增多综合征FIP1L1-PDGFRA融合基因的表达及靶向治疗 总被引:1,自引:0,他引:1
目的研究FIP1L1-PDGFRA融合基因在嗜酸性粒细胞增多综合征(HES)中的表达,评价伊马替尼对HES的靶向治疗作用。方法用荧光原位杂交(FISH)及逆转录-巢式聚合酶链反应(RT-PCR)法检测HES病例F/P融合基因,对阳性标本进行核苷酸序列分析。观察4例伊马替尼治疗患者的临床指标。结果 FISH检测15例标本,1例(8.3%)检测到F/P融合基因。RT-PCR检测24例标本,4例(16.7%)阳性,其中1例用FISH亦检测到F/P融合基因。2例F/P阳性患者,在伊马替尼治疗后短期内均达血液学完全缓解,其中1例达分子生物学缓解。2例F/P阴性者,1例在治疗1月后达血液学完全缓解,另1例于治疗第1天死于急性左心衰。结论 FIP1L1-PDGFRA融合基因是恶性克隆性HES发病的分子基础,伊马替尼治疗F/P阳性患者的血液学缓解率达100%,对部分F/P阴性患者同样有效。 相似文献
28.
急性淋巴细胞白血病8号染色体三体 总被引:2,自引:0,他引:2
目的探讨急性淋巴细胞白血病(ALL)中8号染色体三体(8三体)的发生率。方法对87例ALL和8例正常对照骨髓细胞进行经典的细胞遗传学(CC)及红色荧光素Spectrum Red标记的8号染色体着丝粒α卫星特异DNA探针间期荧光原位杂交技术(FISH)分析。结果87例ALL中14例FISH检测为8三体,占16.09%,5例CC检测结果为8三体(5.75%,5/87);FISH还发现8三体/四体嵌合体1例,而CC仅检测到8四体。结论FISH检测8三体的敏感性高于常规核型分析,在小克隆检测方面有其优越性。 相似文献
29.
Objective To establish and characterize a novel human myeloid leukemia cell line SH-2. Methods Bone marrow mononuclear cells(BMMNC) isolated from a AML-M2 patient, who failed to ob-tain complete remission after chemotherapy and allogenic bone marrow transplantation were passed in a long term IMDM culture medium supplemented with 20% fetal calf serum. Stromal cells were retained and rh-IL-3was added in the culture system. A new human myeloid leukemia cell line SH-2 was successfully established with a cytogeuetic characteristics of a loss of Y chromosome(- Y), a derivative chromosome 16 resulting from unbalanced translocation between chromosome 16 and 17, monosome 17, trisomy 19 and p53 alteration. Vari-ous methods were employed to characterize SH-2 cell line. Results SH-2 cells has been maintained without cytokine and stromal cells for more than 3 years without EB virus and mycoplasma contamination. SH-2 cells had the basically same morphological, immunophenotypic and cytogenetic features as the patient' s leukemia cells did, such as myeloid morphology, an immunophenotype of CD13+ , CD33+ , CD56+ , CD16/56+ and a hypodiploid karyotype of 45, X, - Y, der(16) t(16;17) (q24;q12) , - 17, + 19, which were gradually de-creased and replaced by the near-tetraploid cells with a karyotype of 73 - 102 (80), XX, - Y, - Y, del (lq31) ×2, der(16)t(16;17) (q24;q12) ×2, - 17, - 17, + 19, + 19. FISH and multiple FISH delineated all the abnormalities and revealed a loss of one p53 allele due to monosomy 17. DNA direct sequencing detec-ted a point mutation of CAG to CAT at codon 576 of exon 5 in another p53 allele. RT-PCR showed that SH-2 cells expressed apoptosis-related genes (bcl-2, Fas, GST- π and p21) rather than MDR-related genes. Short tandem repeat PCR provided powerful evidence for the derivation of SH-2 cell line from the patient' s leukemia cells. SH-2 cells had certain colony formation and tumorigenic capacities in nude and SCID mice. Conclu-sion SH-2 is a new myeloid leukemia cell line with a unique biology background, and will provide a useful tool for leukemia research. 相似文献
30.