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脂质体

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Both the increase of lipid peroxidation reaction rate and the shortening of the latent period were dependent on the pre-existed lipid peroxides in liposomes. Al3+ accelerated the oxidation of Fe2+ in liposomal system only when liposomes contain pre-exist peroxides. The addition of Al3+ to the liposomal suspension resulted in a marked increase of turbidity of the suspension and decrease of fluidity of the liposomes.

这可能是因为在脂质体存在的条件下,Al3+加速了Fe2+的氧化,且加速作用与脂质体中原先存在的过氧化物的含量有关;另一方面,Al3+可以引起脂质体的聚集,表现为浊度的增加;测量脂质体上标记的脂肪酸自旋标记物5- Doxyl stearic acid 的ESR 波谱发现: Al3+降低了脂质体的膜脂的流动性。

To observe effect of receptor ligand on subcelluar distribution of liposome was found that alprenolol liposomes was increased in mitochondria and sarcolemma,especially in nuclei(8 fold increased), as compared with blank liposome.

观察受体配基对脂质体在心肌亚细胞器中分布的影响,结果发现,烯丙洛尔掺入脂质体,使脂质体在细胞核、线粒体和肌膜的分布增加,尤其在细胞核增加最明显,较空白脂质体增多了8倍。

Secondly, the thiolation of protein and the coupling of thiol protein with liposomes were studied, and large unilamellar vesicles with highly immunoactivity and stability were acquired.

首先系统研究了混合饱和类脂体系的脂质体的制备和脂质体的性质,然后研究了蛋白质的巯基化和巯基化的蛋白质与脂质体的偶联,从而获得了具有高的免疫活性的、稳定的大单层脂质体

Recombinant plasmid pSVH 7 DNA of avian influenza virus H7 subtype heamagglutinin gene was encapsulated with DC-chol/DOPE liposomes and PC/chol/SA liposomes separately. Two-week old SPF chickens were intramuscularly inoculated with 50 μ g/0.2ml of the liposome entrapped PSVH 7 DNA. Four-weeks later, each chicken was challenged with 0.1ml 〓 AIV . One week after the challenge, the secretion of the cloacas was collected and transfected to chicken embryos to isolate the virus. The virus was isolated from 6/6 of the control group, 1/6 of the naked DNA group, 1/6 of the PC/chol/SA entrapped DNA group and 0/6 of the DC-chol/DOPE liposome entrapped group. The HI antibody titers (log2) of the four groups were 6. 83±0.98, 7. 0±1. 26, 7. 83±1. 17 and 8. 00±0.89 respectively 1-week after challenge, and 8. 5±0.55, 8. 17±0.82, 8. 68±0.45 and 9. 33±0.54 respectively 2-week after challenge. The results showed that inoculation of liposome entrapped DNA significantly enhanced resistance to virosis in animals.

将含禽流感病毒H7亚型血凝素基因的重组质粒pSVH7用DC-chol阳离子脂质体和胆固醇/卵磷脂/十八胺脂质体包裹,免疫2周龄SPF鸡,4周后用同型禽流感病毒进行人工感染,1周后采集泄殖腔分泌物分离病毒,结果未免疫组6/6分离到病毒,裸质粒DNA免疫组1/6分离到病毒胆固醇/卵磷脂/十八胺脂质体包裹DNA免疫组1/6分离到病毒,DC-chol脂质体DNA免疫组没有分离到病毒(0/6):人工感染后1周各组的HI抗体(Log2)分别为6.38±0.98,7.00±1.26,7.83±1.17,8.00±0.89,2周后为8.50±0.55,8.67±0.82,8.68±0.45,9.33±0.52,脂质体包裹组在同期均高于未免疫组和裸DNA免疫组,表明脂质体包裹质粒DNA免疫动物后,能增加动物对病毒感染的抵抗力和反应能力。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体;2)利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达;3)构建ZNRD1的小干扰RNA载体,并测序鉴定;4)利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞(AGS、SGC7901、MKN28)和小鼠成纤维细胞(NIH3T3),G418筛选后进行鉴定;5)利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞(SGC7901)、正常胃组织上皮细胞(GES-1)、对长春新碱耐药的胃癌细胞(SGC7901/VCR)、药敏白血病细胞(HL-60)、对长春新碱耐药的白血病细胞(HL-60/VCR),G418筛选后进行鉴定;6)利用MTT实验检测ZNRD1高/低表达对细胞(AGS、SGC7901、MKN28、NIH3T3、GES-1)生长的影响;7)通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响;8)通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响;9)通过流式细胞仪分析ZNRD1高/低表达对细胞(AGS、MKN28、NIH3T3、GES-1)的细胞周期的影响;10)通过流式细胞仪分析上调ZNRD1对细胞(AGS、MKN28、NIH3T3)的凋亡的影响;11)通过MTT实验检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60、HL-60/VCR)体外药物敏感性的影响;12)通过肾包膜下移植法检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR)体内药物敏感性的影响;13)通过流式细胞仪分析ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60、HL-60/VCR)内阿霉素蓄积和泵出的影响;14)通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响;15)通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60)凋亡敏感性的影响;16)通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响;17)利用RT-PCR、Western blot对基因芯片的结果进行鉴定;18)利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用;19)利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用;20)利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响;21)利用反义核酸技术抑制p21的表达;通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响;22)利用反义核酸技术抑制p27的表达;通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响;23)利用脂质体转染法上调Skp2的表达;通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响;24)利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响;25)利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响;26)利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响;27)利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞(SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR)多药耐药表型的影响;利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

Specifically, itcontains 8 chapters.In chapter 1, the formation, structures, properties and the futureprospect of liposome were thoroughly reviewed;In chapter 2, the stibility and permeability of phopholipid -eleostericacid liposome were studied together with the effect of polymerizationof eleostearic acid. This membrane system was very sensitive to 〓,the effect of 〓 was clarified to increase the aggregation/fusion ofliposomes and made the permeability of mixed liposomes much higher;In chapter 3, two polymerizable conjugated diyne bolaamphiphiles were synthesized. They could form very stable mixed liposome, andthe diyne could be polymerized by UV light in bilayer liposomes, as aresult, the stability of mixed liposome against solvent or surfactantafter polymerization were enhanced. In chapter 4, two kinds of amphiphilic amino acids were synthesized andstable liposomes were formed therefrom After the condensationpolymerization of amino acid in bilayer liposomes, stable polypeptide liposomes were obtained, which had lower phase transition temperatureand higher permeability.In chapter 5, four kinds of glycolipids were synthesized and theiraggregation behavior in water was comparied. When incorporated intophospholipid bilayer membranes, they could increase the phase transitiontemperatures and inhibit the aggregation and fusion of mixedliposomesat lower temperature.In chapter 6 and 7, three kinds of steroidal bolaamphiphiles withdifferent chain lengths were synthesized. Incorporation of steroidalmoiety to the center of lipid bilayer membrane obviously increased themobility of lipid membrane and shifted Tc to lower temperature side incomparasion with cholesterol. The bolaamphiphile which was shorter thanthe hosted lipid bilayer membrane thickness influenced the lipid packingmore obviously.

全文共分8章:第一章对脂质体的形成、结构、性质及展望进行了较为详细的文献综述;第二章研究了磷脂-桐酸脂质体的稳定性,通透能力及桐酸的聚合对这些性质的影响;磷脂-桐酸混合脂质体为一类对〓灵敏的脂质体,〓的作用首先是使脂质体集聚然后使脂质体融合,并加速内包荧光物的释放;第三章通过合成两种可聚合共轭双炔双极性双亲分子DDCA,DDOL,研究了共双炔分子在双分子层脂质体膜上的聚合及对脂质体性质的影响,聚合可以提高脂质体相对于溶剂及表面活性剂的稳定性;第四章合成了两类氨基酸为极性基团的双亲分子,它们均可以在超声下形成稳定的脂质体结构;氨基酸基团可以在脂质体上进行缩聚反应,若聚合后脂质体表面仍有足够的亲水能力,则可得到稳定的多肽型脂质体;聚合后脂质体的相变温度降低,通透能力增加;第五章合成了四种亲水基团为单糖基的双亲分子GL-l,GL-2,GL-3, GL-4,研究了它们在水中的分散情况、集合体形态与分子结构的关系;在DMPC双分子层膜中加入糖脂分子可以使脂质体的相变温度提高,阻止脂质体在低温放置时的集聚与融合;第六章-第七章合成了三种不同碳链长度的双极性含胆甾环双亲分子 CL-1,CL-2,CL-3;它们可以象胆固醇一样与磷脂混合形成稳定脂质体,胆甾环基团位于脂质体双分子层膜的中间;与胆固醇的作用相反,它们可以增加磷脂双分子层膜的流动性,降低混合脂质体的相变温度;三种分子的作用与其碳链长度和磷脂双分子层膜的厚度有关,比膜厚度短的分子影响最大。

Liposome properties and biological behaviors can be controlled by surface modification. Attachments of hydrophilic polymers, such as poly, enable them to have a prolonged circulation time without being opsonized and recognized by mononuclear phagocytic system.

通过表面修饰可以改变脂质体的性质和生物学行为:在脂质体表面修饰亲水性高分子可以阻碍血浆成分的调理作用,减少脂质体被网状内皮系统识别和摄取,使脂质体具有长循环的特点;在脂质体表面修饰特异性的配基,能使脂质体具有主动靶向的特性。

Almost all CF was released from Dansyl-PE by CTX-IX that osmosed the Dansyl-PE liposome like detergent; CF releases from PE or PS liposome induced by CTX-IX were related with both of CTX-IX and phospholipid concentrations, this indicated that the phase seperation should be a reasonable mechanism for CF permeability of PS or PE liposome in the presence of CTX-IX rather than CTX-IX channel formation on the membrane. CTX-IX influence of CF release of PC liposome was very slight. CF release from Dansyl-PE、PE or PE liposome in response to CTX-IX could be inhibited by Ca〓.

CTX-IX对靶脂如此种种强烈的作用,使得它引起含Dansyl-PE脂质体CF的泄漏犹如击污剂那样强烈;引起含PS或PE脂质体CF渗透的增加为一个与CTX-IX及磷脂浓度有关的二级反应过程,这种动力学特征提示渗透性增加的机制可能是CTX-IX在脂膜上产生的脂质分子相分离,而不是CTX-IX脂膜上形成通道;CTX-IX对PC脂质体渗透性几乎没有影响,而对上述脂质体渗透性增加作用均可被Ca〓所抑制。

Including the soybean lecithin separation purification, gamma - the linolenic acid separation and the purification, as well as gamma - linolenic acid fat 质体 Bao He studies and so on, and carries on optimal using the orthogonal experiment to the above craft.

其中包括大豆卵磷脂的分离纯化,γ-亚麻酸的分离和纯化、以及γ-亚麻酸脂质体包合研究等等,并利用正交试验对上述工艺进行优选。

East valuable Jin Danzhong the gamma - linolenic acid and the soybean lecithin will make the fat 质体 both to conform to the original group side principle and to be possible to increase the preparation the stability and the curative effect.

将东宝金丹中γ-亚麻酸和大豆卵磷脂制成脂质体既符合原组方原则又可增加制剂的稳定性和疗效。

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The labia have now been sutured together almost completely.The drains and the Foley catheter come out at the top.

此刻阴唇已经几乎完全的缝在一起了,排除多余淤血体液的管子和Foley导管从顶端冒出来。

To get the business done, I suggest we split the difference in price.

为了做成这笔生意,我建议我们在价格上大家各让一半。

After an hour and no pup, look for continued contractions and arching of the back with no pup as a sign of trouble.

一个小时后,并没有任何的PUP ,寻找继续收缩和拱的背面没有任何的PUP作为一个注册的麻烦。