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Objective:In order to confirm the tumorigenecity of Epstein-Barr virus(EBV)to human normal cells by the test in animal body and to detect the Chinese susceptibility to E-BV in fection,the animal model of EBV-induced tumor has been established.Methods:Human peripheral blood lymphocytes(PBLs)were trasplantesd intraperitoneally into severely combined im-munodeficient (SCID)mice by1x10PBLs per mouse.The mice with hu-PBL engraftment from healthy IgA/VCA negative donors were injected intraperitoneally in a week...

Objective:In order to confirm the tumorigenecity of Epstein-Barr virus(EBV)to human normal cells by the test in animal body and to detect the Chinese susceptibility to E-BV in fection,the animal model of EBV-induced tumor has been established.Methods:Human peripheral blood lymphocytes(PBLs)were trasplantesd intraperitoneally into severely combined im-munodeficient (SCID)mice by1x10PBLs per mouse.The mice with hu-PBL engraftment from healthy IgA/VCA negative donors were injected intraperitoneally in a week with EBV-containing supernatant from B95-8cell suspension culture(active infection),whereas the mice receiving hu-PBL engraftment from healthy IgA/VCA positive donors were not injected with B95-8supernatant (latent infection).Results:In19hu-PBL/SCID chimerical recipients derived from12donors,the tumor incidences were91.7%for12cases of donor(11/12)and84.2%for19survival SCID mice(16/19)respectively.Another12animals died because of graft-versus-host disease and their average life span was17.5days.EBV-induced tumors were often found in the abdominal cavity or mediastinum which possessed aggressive and fatal features.The average survival time of tumor-bearing mice was65.5days.The EBV-induced neoplasms in SCID mice were nodular tumors.Microscopic observation of tumors showed diffuse arrangement.Histologically the tumors were dominatedy by large-cleaved and non-cleaved mixed cells.Im munohistochemical analysis of tu mors in mice re vealed that LCA(CD45)and L26(human B-cell marker CD20)were positive,but both PS1and UCHL-1(T-cell marker,CD3and CD4RD)were negative.Observation of histopathology and im munopatholo gy demonstrated that the induced tu mors were human B-cell lym phomas.In situ hybridization revealed that the tumor cells contained the EBV encoded small RNA-1,i.e.E-BER-1and Alu sequence of human genome.There were EBV particles in Nuclei of tumor cells ob served by electron mi croscopy.Immunohistochemistry showed positive stain ing of BZLF-1protein in almost all tumor cells.Conclusion:Human PBL/SCID chimeras is a sensitive animal model to study infection and oncogenesis of EB vitrus.This experiment has ob tained the direct causative evidence dealing with EBV associated tumor derived from human normal cells.In the present work,the findings confirmed the oncogenecity of EB virus to human normal cells and defined sig nificant role of the immunodeficiency in the process of tumor develop ment.The re sults suggest the high susceptibility of South Chinese to EBV infection.

目的:通过动物体内试验研究EB病毒(EBV)对人正常细胞的致瘤性,并检测我国正常人群对EBV的易感性,试图建立EBV诱发人淋巴细胞肿瘤模型。方法:在严重联合免疫缺陷动物即SCID小鼠体内移植健康成人外周血淋巴细胞(PBL),每鼠腹腔接种1×108个PBL。对接种VCA/IgA阴性献血员PBL的动物,移植后1周内经腹腔注射B95-8标准株EBV悬液作为实验感染,而接种VCA/IgA阳性献血员PBL的动物不再进行实验感染。结果:在接受12名健康成人PBL移植并感染EBV的19只SCID小鼠中,肿瘤诱发率分别为91.7%(11/12名)和84.2%(16/19只鼠)。诱发瘤常见于小鼠腹腔后壁和纵隔,具有侵袭性和致死性,患瘤小鼠平均存活时间65.5天。EBV诱发瘤是结节状实体瘤,显微镜下观察肿瘤细胞呈大裂—无裂混合细胞。组织病理学和免疫病理学研究表明,肿瘤类型是人源B细胞性恶性淋巴瘤。诱发瘤原位分子杂交显示肿瘤细胞核内存在EB病毒小核酸分子EBER-1和人类基因组Alu序列。电子显微镜观察肿瘤细胞核内存在EB病毒颗粒。肿瘤细胞表达EB病毒BZLF1蛋白阳性。结论:SCID/人淋巴细胞嵌合体是研究EBV感染...

目的:通过动物体内试验研究EB病毒(EBV)对人正常细胞的致瘤性,并检测我国正常人群对EBV的易感性,试图建立EBV诱发人淋巴细胞肿瘤模型。方法:在严重联合免疫缺陷动物即SCID小鼠体内移植健康成人外周血淋巴细胞(PBL),每鼠腹腔接种1×108个PBL。对接种VCA/IgA阴性献血员PBL的动物,移植后1周内经腹腔注射B95-8标准株EBV悬液作为实验感染,而接种VCA/IgA阳性献血员PBL的动物不再进行实验感染。结果:在接受12名健康成人PBL移植并感染EBV的19只SCID小鼠中,肿瘤诱发率分别为91.7%(11/12名)和84.2%(16/19只鼠)。诱发瘤常见于小鼠腹腔后壁和纵隔,具有侵袭性和致死性,患瘤小鼠平均存活时间65.5天。EBV诱发瘤是结节状实体瘤,显微镜下观察肿瘤细胞呈大裂—无裂混合细胞。组织病理学和免疫病理学研究表明,肿瘤类型是人源B细胞性恶性淋巴瘤。诱发瘤原位分子杂交显示肿瘤细胞核内存在EB病毒小核酸分子EBER-1和人类基因组Alu序列。电子显微镜观察肿瘤细胞核内存在EB病毒颗粒。肿瘤细胞表达EB病毒BZLF1蛋白阳性。结论:SCID/人淋巴细胞嵌合体是研究EBV感染和致瘤性的敏感动物模型,且获得了EBV引起人类正常细胞在体内发生肿瘤的直接依据。因而证实了EBV对人类细胞的致瘤作用,进一步明确机体免疫缺陷因素在肿瘤发生中起重要作?

Objective To elucidate the killing activity of yeast cytosine deaminase /5-fluorocytosine (YCD/5-FC) gene therapy system on gene-transferred tumorigenic cell line K562B in vivo. Method K562B cell was infected with high titer virus and a gene transferred cell clone,YCD-K562B,was selected. Twelve male SCID mice of 4 week old were divided into 2 groups at random and both YCD-K562B and K562B cells were implanted to each mice. 5-FC or saline was given i.p for 10 days after tumor developed, and relative tumor...

Objective To elucidate the killing activity of yeast cytosine deaminase /5-fluorocytosine (YCD/5-FC) gene therapy system on gene-transferred tumorigenic cell line K562B in vivo. Method K562B cell was infected with high titer virus and a gene transferred cell clone,YCD-K562B,was selected. Twelve male SCID mice of 4 week old were divided into 2 groups at random and both YCD-K562B and K562B cells were implanted to each mice. 5-FC or saline was given i.p for 10 days after tumor developed, and relative tumor volume was measured every 3 days. At the end of experiment, animals were sacrificed and the specimens were processed for histopathological examination. Results At the end of experiment (21 days after tumor cell implantation), the relative tumor volume of the 4 groups were:YCD-K562B+5-FC 2.922±0.581,YCD-K562B+saline 24.434±4.790,K562B+5-FC 22.701±2.350 and K562B+saline 24.460±1.670;t-test analysis showed that 5-FC could kill cells (YCD-K562B)in vivo(P=0.000?1),but had no effect on the growth of gene-untransferred cells (K562B) (P=0.096). In YCD-K562B+5-FC group, relative tumor volume reduced in 3~6 days after treatment(the minimum was 0.681). Necrosis around artery could be found in the tumor of YCD-K562B+5-FC group. Conclusion YCD/5-FC suicide gene therapy system has a significant in vivo killing activity to gene-transferred tumorigenic YCD-K562B cell.

目的 了解酵母菌胞嘧啶脱氨酶 5 氟胞嘧啶 (YCD 5 FC)系统在体内对转基因高致瘤性K5 6 2细胞 (K5 6 2B细胞 )的杀伤效应。方法 以高滴度逆转录病毒转染K5 6 2B细胞并筛选出阳性转染克隆YCD K5 6 2B ;12只SCID小鼠分为治疗及对照组 ,在小鼠左右两侧近前肢处腹部皮下注射YCD K5 6 2B及K5 6 2B细胞 ,成瘤后治疗组腹腔注射 5 0 0mg kg 5 FC共 10d ,对照组腹腔注射生理盐水 ,观察瘤体相对体积变化及病理变化。结果 瘤细胞接种第 2 1天 ,瘤体相对体积分别为 :YCD K5 6 2B +5 FC组 2 .92 2± 0 .5 81,YCD K5 6 2B +生理盐水组 2 4.434± 4.790 ,K5 6 2B +5 FC组 2 2 .70 1± 2 35 0 ,K5 6 2B +生理盐水组 2 4.46 0± 1.6 70 ;YCD K5 6 2B +5 FC组与YCD K5 6 2B +生理盐水组相比差异非常显著 (P =0 0 0 0 1) ,K5 6 2B +5 FC组及K5 6 2B +生理盐水组相比...

目的 了解酵母菌胞嘧啶脱氨酶 5 氟胞嘧啶 (YCD 5 FC)系统在体内对转基因高致瘤性K5 6 2细胞 (K5 6 2B细胞 )的杀伤效应。方法 以高滴度逆转录病毒转染K5 6 2B细胞并筛选出阳性转染克隆YCD K5 6 2B ;12只SCID小鼠分为治疗及对照组 ,在小鼠左右两侧近前肢处腹部皮下注射YCD K5 6 2B及K5 6 2B细胞 ,成瘤后治疗组腹腔注射 5 0 0mg kg 5 FC共 10d ,对照组腹腔注射生理盐水 ,观察瘤体相对体积变化及病理变化。结果 瘤细胞接种第 2 1天 ,瘤体相对体积分别为 :YCD K5 6 2B +5 FC组 2 .92 2± 0 .5 81,YCD K5 6 2B +生理盐水组 2 4.434± 4.790 ,K5 6 2B +5 FC组 2 2 .70 1± 2 35 0 ,K5 6 2B +生理盐水组 2 4.46 0± 1.6 70 ;YCD K5 6 2B +5 FC组与YCD K5 6 2B +生理盐水组相比差异非常显著 (P =0 0 0 0 1) ,K5 6 2B +5 FC组及K5 6 2B +生理盐水组相比 ,差异无显著性 (P =0 .0 96 ) ,表明 5 FC对转YCD基因的K5 6 2B白血病细胞有明显的杀伤效应 ,而对未转基因细胞的生长无影响 ;YCD K5 6 2B +5 FC组瘤体于瘤细胞接种后第 12~第 15天 (5 FC治疗的第 3~第 6天 )有所缩小 (最小的相对体积为 0 .6 81) ,病理检查可见 5 FC治疗组瘤体有以小动脉血管为中心的坏死。结论 YCD 5 FC系统在体内对转YCD

Objective:To study the effect of Cyclosporine A(CSA) on inhibiting graft versus host reaction(GVHR) occured in hu PBL/SCID chimeras and to stably establish EBV induced lymphoma models.Methods:Human peripheral blood lymphocyts were isolated and were inoculated intraperitoneally into SCID mice.Mice were infected with EBV and injected intraperitoneally with CSA.Human sIL 2R in the serum of hu PBL/SCID chimeras were analyzed by ELISA.Results:No mouse was dead in CSA group,whereas 15 mice of the other three groups...

Objective:To study the effect of Cyclosporine A(CSA) on inhibiting graft versus host reaction(GVHR) occured in hu PBL/SCID chimeras and to stably establish EBV induced lymphoma models.Methods:Human peripheral blood lymphocyts were isolated and were inoculated intraperitoneally into SCID mice.Mice were infected with EBV and injected intraperitoneally with CSA.Human sIL 2R in the serum of hu PBL/SCID chimeras were analyzed by ELISA.Results:No mouse was dead in CSA group,whereas 15 mice of the other three groups died of GVHR.The medium life span of no CSA administration mice was 17 days,and motalities were 55.56%(5/9),30.43%(7/23),42.86%(3/7)respectively.The difference was statistically significant between CSA group and the other groups.The levels of human sIL 2R were stable in CSA group while increased gradually in experimental infertion the groups without CSA.Difference was significant at day 15 and day 22 between the EBV infection group without CSA and with CSA administration.Of 38 survival SCID mice,24 mice developed tumors in their body cavities.Conclusion:CSA can strikingly inhibit GVHR that may occur in hu PBL/SCID mice,that could help practical to stably establish the lymphoma models.

目的 :观察环孢素A(CSA)对huPBL SCID嵌合体小鼠发生移植物和抗宿主反应 (GVHR)的抑制作用 ,建立稳定的EBV诱发淋巴瘤动物模型。方法 :从健康成人新鲜外周血中分离出淋巴细胞 ,将之移植到SCID小鼠腹腔中 ,实验感染EB病毒 ,腹腔注射CSA ,并采用ELISA检测小鼠血清中人sIL 2R水平。结果 :环孢素A组 (14只 )无 1只小鼠因GVHR而死亡 ,而其余 3组共 39只小鼠有 15只因GVHR而死亡 ,中位生存时间为 17d ,死亡率分别为 5 5 5 6 % (5 9例 )、30 4 3% (7 2 3例 )、4 2 86 %(3 7例 ) ,与环孢素A组比较差异有显著性意义。环孢素A组在不同时间sIL 2R含量较稳定 ,而实验感染组sIL 2R水平有逐渐升高趋势 ,环孢素A组与实验感染组同一时间比较 ,15d和 2 2d时sIL 2R水平有显著性差异 ,后者明显高于前者。渡过急性GVHR期而存活的 38只SCID小鼠中 ,共有 2 4只形成肿瘤。结论 :CSA可显著抑制hu PBL SCID嵌合体小鼠GVHR的发生 ,对稳定建立EBV诱发淋巴瘤动物模型具有...

目的 :观察环孢素A(CSA)对huPBL SCID嵌合体小鼠发生移植物和抗宿主反应 (GVHR)的抑制作用 ,建立稳定的EBV诱发淋巴瘤动物模型。方法 :从健康成人新鲜外周血中分离出淋巴细胞 ,将之移植到SCID小鼠腹腔中 ,实验感染EB病毒 ,腹腔注射CSA ,并采用ELISA检测小鼠血清中人sIL 2R水平。结果 :环孢素A组 (14只 )无 1只小鼠因GVHR而死亡 ,而其余 3组共 39只小鼠有 15只因GVHR而死亡 ,中位生存时间为 17d ,死亡率分别为 5 5 5 6 % (5 9例 )、30 4 3% (7 2 3例 )、4 2 86 %(3 7例 ) ,与环孢素A组比较差异有显著性意义。环孢素A组在不同时间sIL 2R含量较稳定 ,而实验感染组sIL 2R水平有逐渐升高趋势 ,环孢素A组与实验感染组同一时间比较 ,15d和 2 2d时sIL 2R水平有显著性差异 ,后者明显高于前者。渡过急性GVHR期而存活的 38只SCID小鼠中 ,共有 2 4只形成肿瘤。结论 :CSA可显著抑制hu PBL SCID嵌合体小鼠GVHR的发生 ,对稳定建立EBV诱发淋巴瘤动物模型具有重要的实际意义。

 
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