排序方式: 共有71条查询结果,搜索用时 15 毫秒
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乙型肝炎病毒表面抗原(HBsAg)基因重组杆状病毒的构建及鉴定 总被引:1,自引:0,他引:1
目的 利用Bac-to-Bac杆状病毒表达系统,构建乙型肝炎病毒(HBV)表面抗原基因的重组杆状病毒。方法 构建含有HBV S基因的表达截体pFBDHTa—S,转化DH10Bac感受态菌。利用其含有的细菌Tn7转座系统,将该基因重组至杆状病毒穿梭质粒bacmicl中,转染昆虫细胞Sf9,获得含有HBV S基因的重组杆状病毒。结果 构建了的含HBV S基因的重组杆状病毒,感染昆虫细胞后,PCR检测可以扩增出相应大小的片段。结论 利用杆状病毒表达系统,成功地构建了含HBV S基因的重组杆状病毒,为进一步的研究奠定了基础。 相似文献
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由侧脑室或在中脑导水管周围灰质内微量注射入白细胞α-干扰素可以引起大鼠痛阈明显上升,但脑内5-HT和5-HIAA均无变化。看来干扰素的镇痛作用和脑内5-HT递质无关,它可能是不经过5-HT的释放也不和吗啡受体结合起作用的另一类型的镇痛物质。 相似文献
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胃癌浸润性淋巴细胞的细胞表型及杀伤活性 总被引:1,自引:0,他引:1
目的和方法:本实验用9例胃癌患者TIL和IL2在体外共同孵育后,用流式细胞仪分析胃癌TIL的细胞表型特征。结果:TIL细胞表型特征是以CD3为主,CD4/CD8为083,NK细胞163±36%。经白细胞介素2(IL2)激活20天后,CD3减少,CD4/CD8为185,NK细胞数量增至413±137%,P<001。LDH释放法测定激活后的TIL对自体胃癌细胞杀伤率比培养初期高374倍,变比对同时培养的7901胃癌细胞杀伤率高175倍。对自体和异体胃癌细胞杀伤作用均显著高于LAK细胞,且具有靶细胞特异性。结论:结果表明胃癌TIL对胃癌细胞杀伤作用,可能与NK细胞数量增多,导致胃癌细胞凋亡有关 相似文献
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目的 建立一套快速、灵敏和特异的人抗黏液蛋白A(MxA)基因启动子中干扰素刺激应答元件-88/-123位多态性检测体系,为合理应用IFN-α治疗慢性乙型肝炎提供分子生物学检测手段.方法 对经IFN-α治疗的患者进行HBV DNA、HBV基因分型及MxA基因启动子中干扰素刺激应答元件-88/-123位多态性位点检测,确定基因多态性与IFN-α治疗效果之间的关系.通过各种条件优化实验,建立MxA基因启动子中干扰素刺激应答元件-88/-123位多态性荧光PCR检测体系,并通过与DNA序列分析法检测结果的对比,验证荧光PCR检测体系的灵敏性及特异性,从而对该体系的临床适用性进行初步评估.采用卡方检验计算P值、回归分析法计算对比率和95%可信区间.结果 MxA基因启动子干扰素刺激应答元件中-88位为G/T杂合型和-123位为C/A杂合型者皆为IFN-α敏感型,-88位为G/G纯合型和-123位为C/C纯合型者为IFN-α不敏感型.与DNA序列分析的金标准相比,荧光PCR检测的符合率为99.65%.结论 荧光PCR检测体系,可灵敏、快捷地检测患者MxA基因多态性位点. 相似文献
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成人睾丸组织在裸鼠体内的存活情况研究 总被引:1,自引:0,他引:1
目的以成人睾丸组织为供体,免疫缺陷小鼠为受体,研究成人睾丸组织中生精细胞异种移植后的存活及变化情况。方法将成人正常睾丸组织植入去势裸鼠背部,于移植后110d取出移植物,进行组织形态学观察。结果在一例移植前显示非正常生精上皮结构标本的移植物中,大多数生精小管中只含有Sertoli细胞,而另一例移植前显示出正常生精上皮结构的标本,在移植到裸鼠背部110d后,仍然观察到圆形和长形精子细胞存在。结论将成人睾丸组织移植到免疫缺陷小鼠体内,110d后仍观察到有生精细胞的存活。 相似文献
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Objective: To evaluate the differentiation of human umbilical cord blood cells into hepatocyte-like cells. Methods: Mononuclear cells (MNCs) derived from human umbilical cord blood were isolated using Ficoll. The experiment was derived into 3 categories:(1) MNCs co-cultured with 50 mg minced liver tissue separated by a trans-well membrane and then collected at 0 h,24 h,48 h and 72 h; (2) MNCs cultured along supplemented with 100 ml/L FBS. 100μ/ml penicillin, 100μg/ml streptomycin. 4. 7μg/ml linoleic acid, 1×ITS, 10-4 mol/L L-Ascorbic acid 2-P and a combination of FGF4 (100 ng/ml) and HGF (20 ng/mL). Cells were then collected at 0 d and 16 d to examine the expression profile of hepatocyte correlating markers;(3) 0. 2-0. 3 ml of MNCs with a cell density of 2×107/ml were transplanted into prepared recipient mice [n = 12, injected with 0. 4 ml/kg (20%) CCl4 and 150 ng/kg 5-fluorouracil (5-Fu) prior the transplant 24 h and 48 h, respectively] via injection through tail vein. Mice were sacrificed 4 weeks after transplantation. The hepatocyte correlating mRNAs and proteins were determined by RT-PCR, immunohistochemical analysis and immunoflurence technique. Results: (1) After 72 h. a number of glycogen positive stained cells were observed with MNCs co-cultured with damaged mouse liver tissues. The expression of hepatocyte markers, human albumin (ALB),α-fetal protein (AFP) and human GATA4 mRNA and proteins were detected by RT-PCR and immunohistochemistry as well. For the confirmation, the DNA sequencing of PCR products was performed. In control groups, MNCs co-cultured with normal mouse hepatocytes or MNCs cultured alone, all markers remained negative. (2) In growth factor supplemented culture system, MNCs developed into larger volume with richer cytoplasm and binucleation after 16 d. Positive expression of ALB, AFP, CK18 and CK19 mRNA were detected with RT-PCR, and ALB positive staining was observed by immunocytochemistry as well. In contrast. MNCs cultured without exogenous growth factors scarcely attached to the culture dish and ALB mRNA was not detected. (3) In transplantation experiment, both of ALB and AFP mRNA were detected by RT-PCR and HSA, PCNA and ALB positive staining were observed in the livers of recipient mice by immunocytochemistry. Conclusion: MNCs from human umbilical cord blood could convert into hepatocyte-like cells in 3 different ways, indicating their potential use in the clinic applications for the treatment of human liver diseases. 相似文献
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脐血CD34+细胞的分离与纯化 总被引:2,自引:0,他引:2
脐血造血干细胞的分离是进行干细胞移植、体外扩增培养的关键.研究采用明胶自然沉降法、Fi-coll分层法分离脐血MNC后,用MACS分离纯化CD34+细胞,计数并用流式细胞仪进行纯度分析.结果显示每份脐血的采集量平均为95±52 ml,3 g/dl明胶自然沉降法所获MNC密度平均为(5.76±0.67)×107/ml;CD34+细胞密度平均为(5.53±1.16)×105/ml,较Ficoll分层法高(P<0.01).因此,3 g/dl明胶自然沉降法是一种较理想的分离MNC的方法,用明胶沉降法和MACS相结合是分离脐血CD34+细胞的理想方法. 相似文献