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1.
为提高人β-珠蛋白基因在基因转移中的稳定性与在红系细胞中的表达水平,构建了带单个和多个基因座控制区核心序列的人β-珠蛋白基因相关病毒(adeno-associatedvirus,AAV)载体AV53HS2△β2Neo和AV53HS432△β2Neo。利用AAV载体介导的基因转移将两种重组体导入红系细胞(MEL),分析了它们在MEL细胞中的整合与表达。结果表明,AAV载体介导带多个基因座控制区核心序  相似文献   

2.
目的 利用腺相关病毒9(AAV9)介导的CRISPR/Cas9系统构建成体心肌细胞特异性Oga基因敲除小鼠模型,来研究内源性OGA在成体心脏稳态维持中的功能。方法 首先,筛选靶向Oga编码区的有效sgRNA,构建包装表达有效sgRNA的AAV9病毒。其次,建立心肌细胞特异性SpCas9表达小鼠(α-MHCCas9),并通过腹腔注射方式将AAV9-sgOga注射到小鼠体内。通过qPCR和Western blot印记杂交实验检测Oga的表达情况,确定Oga敲除是否成功。最后,通过组织学分析心脏结构,以及超声心动图分析心脏功能,来分析Oga对心脏稳态维持的影响。结果 筛选到2条可以有效靶向Oga编码区的sgRNA,通过AAV9递送实现了Oga基因在心脏组织的有效敲除,且导致O-GlcNAcylation表达显著上升;组织学分析和超声心动图分析发现基因敲除小鼠心脏结构及功能在基础水平与对照小鼠无显著变化。结论 利用AAV9介导的CRISPR/Cas9系统成功建立了成体心肌细胞Oga基因敲除的小鼠模型,为研究OGA介导的O-GlcNAcylation清除在心脏稳态维持中的功...  相似文献   

3.
Hemophilia B is a hemorrhagic disease resulting from Factor Ⅸ gene (hFⅨ) mutation as an X-linked recessive inherited trait. The incidence of this disease is 1 in 30000. Clinical treatments depend mainly upon blood transfusions or administration of prothrombin complex so that patients are at the risk of infections with the HIV and hepatitis viruses. Gene therapy offers an attractive alternative in the treatment of hemophilia B by eliminating those risks. In 1991, our lab conducted clinica…  相似文献   

4.
血友病性关节病(HA)的发病机制与关节内炎性细胞因子过表达有关,肿瘤坏死因子α(TNFα)是最重要的炎性因子之一。可溶性肿瘤坏死因子受体(sTNFR)作为TNFα的受体可以特异性地与TNFα结合拮抗TNFα炎性效应。为探讨关节腔内持续表达sTNFR对HA关节的保护作用,将表达TNFR的质粒(pAAV-TNFR:Fc)包装为rAAV5-TNFR:Fc载体,注射到血友病小鼠膝关节内。结果显示炎症持续42 d后,关节内仍能表达TNFR:Fc,两个给药组中TNFα和白介素-1β(IL-1β)的表达不同程度地下降,关节病变均显著减轻,表明该关节内基因治疗对血友病性关节病有预防和治疗效果。  相似文献   

5.
腺相关病毒(Adeno-associated virus-2,AAV-2)用于构建基因治疗病毒载体,在基因治疗领域受到了普遍重视和关注,而rAAV在应用过程中一个很重要的限制因素就是缺乏简单有效的大规模制备方法.运用昆虫细胞表达系统来提高AAV-2的产量,分别采用绿色荧光蛋白(GFP)和胰岛素(Insulin)基因替代AAV-2的结构基因后,与其他重组病毒共转染293t细胞,3 d后即可检测到高效表达的GFP和Insulin.结果表明,该方法能够比较理想地提高AAV的产量,为临床使用奠定了基础.  相似文献   

6.
《矿物冶金与材料学报》2021,28(12):2001-2007
Graphene oxide (GO) wrapped Fe3O4 nanoparticles (NPs) were prepared by coating the Fe3O4 NPs with a SiO2 layer, and then modifying by amino groups, which interact with the GO nanosheets to form covalent bonding. The SiO2 coating layer plays a key role in integrating the magnetic nanoparticles with the GO nanosheets. The effect of the amount of SiO2 on the morphology, structure, adsorption, and regenerability of the composites was studied in detail. An appropriate SiO2 layer can effectively induce the GO nanosheets to completely wrap the Fe3O4 NPs, forming a core-shell Fe3O4@SiO2@GO composite where Fe3O4@SiO2 NPs are firmly encapsulated by GO nanosheets. The optimized Fe3O4@SiO2@GO sample exhibits a high saturated adsorption capacity of 253 mg·g?1 Pb(II) cations from wastewater, and the adsorption process is well fitted by Langmuir adsorption model. Notably, the composite displays excellent regeneration, maintaining a ~90% adsorption capacity for five cycles, while other samples decrease their adsorption capacity rapidly. This work provides a theoretical guidance to improve the regeneration of the GO-based adsorbents.  相似文献   

7.
《矿物冶金与材料学报》2021,28(12):1908-1916
The effect of CaCO3, Na2CO3, and CaF2 on the reduction roasting and magnetic separation of high-phosphorus iron ore containing phosphorus in the form of Fe3PO7 and apatite was investigated. The results revealed that Na2CO3 had the most significant effect on iron recovery and dephosphorization, followed by CaCO3, the effect of CaF2 was negligible. The mechanisms of CaCO3, Na2CO3, and CaF2 were investigated using X-ray diffraction (XRD), scanning electron microscopy and energy dispersive spectrometry (SEM–EDS). Without additives, Fe3PO7 was reduced to elemental phosphorus and formed an iron–phosphorus alloy with metallic iron. The addition of CaCO3 reacted with Fe3PO7 to generate an enormous amount of Ca3(PO4)2 and promoted the reduction of iron oxides. However, the growth of iron particles was inhibited. With the addition of Na2CO3, the phosphorus in Fe3PO7 migrated to nepheline and Na2CO3 improved the reduction of iron oxides and growth of iron particles. Therefore, the recovery of iron and the separation of iron and phosphorus were the best. In contrast, CaF2 reacted with Fe3PO7 to form fine Ca3(PO4)2 particles scattered around the iron particles, making the separation of iron and phosphorus difficult.  相似文献   

8.
9.
Carbonated decomposition of hydrogarnet is one of the vital reactions of the calcification–carbonation method, which is designed to dispose of low-grade bauxite and Bayer red mud and is a novel eco-friendly method. In this study, the effect of the silica saturation coefficient (x) on the carbonation of hydrogarnet was investigated from the kinetic perspective. The results indicated that the carbonation of hydrogarnets with different x values (x = 0.27, 0.36, 0.70, and 0.73) underwent two stages with significantly different rates, and the kinetic mechanisms of the two stages can be described by the kinetic functions R3 and D3. The apparent activation energies at Stages 1 and 2 were 41.96–81.64 and 14.80–34.84 kJ/mol, respectively. Moreover, the corresponding limiting steps of the two stages were interfacial chemical reaction and diffusion.  相似文献   

10.
《矿物冶金与材料学报》2020,27(10):1347-1352
A new method of high-gravity combustion synthesis (HGCS) followed by post-treatment (PT) is reported for preparing high-performance high-entropy alloys (HEAs), Cr0.9FeNi2.5V0.2Al0.5 alloy, whereby cheap thermite powder is used as the raw material. In this process, the HEA melt and the ceramic melt are rapidly formed by a strong exothermic combustion synthesis reaction and completely separated under a high-gravity field. Then, the master alloy is obtained after cooling. Subsequently, the master alloy is sequentially subjected to conventional vacuum arc melting (VAM), homogenization treatment, cold rolling, and annealing treatment to realize a tensile strength, yield strength, and elongation of 1250 MPa, 1075 MPa, and 2.9%, respectively. The present method is increasingly attractive due to its low cost of raw materials and the intermediate product obtained without high-temperature heating. Based on the calculation of phase separation kinetics in the high-temperature melt, it is expected that the final alloys with high performance can be prepared directly across master alloys with higher high-gravity coefficients.  相似文献   

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