英语翻译2.2.Nanoparticle preparationNanoparticles were prepared

英语翻译
2.2.Nanoparticle preparation
Nanoparticles were prepared and DNA was incorporated according to the method described by Duceppe and Tabrizian(2009).In brief,10 mg HA was dissolved in 4 ml of buffer (0.1 M sodium acetate,0.1 M sodium chloride,pH7.2) under magnetic stirring.After HA was completely dissolved,0.5mg hyaluronidase (from bovine testes,activity 300–500 units/mg,Sigma–Aldrich,USA) was added to the solution,which was then kept for 24 h in a shaking incubator at 37?C.After the enzymatic splitting reaction,the mixture was filtered (Amicon Ultra,MWCO 10kDa,Millipore Corp.,USA),and then the filtrated HA of low molecular weight (MW < 10 kDa) was centrifuged and collected by lyophilization (de laFuenteetal.,2008).The resultingsubstance was dissolved in distilled water (pH 5.5).10 mg of CS was dissolved in 2% acetic acid (pH5.5).Both the HA and CS solutions were filtered through a 0.22 m membrane.The CS solution was stirred at a rate of 3000 rpm for 30 min,mixed into the HA solution and stirred for 10 min.Eight different mixtures were prepared with CS:HA weight ratios at:1:2,1:1,2:1,3:1,4:1,5:1,6:1,and 7:1.A constant concentration of 11.25 g/ml of HA was used in all mixtures,and the CS concentration was varied as:5.625,11.25,22.5,33.75,45,56.25,67.5,and 78.25 g/ml.The required volume of 25 g/ml plasmid DNA was gently added to CS/HA solution by gentle pipetting to form complexes of a selected N/P ratio.N/P ratio was defined as the molar ratio of the positive CS amino group and the negative DNA phosphate group.The mixture was vortexed rapidly for 3–5 s and left for 1 h at room temperature for the complexes to completely form.

纳米preparationNanoparticles准备和DNA成立根据描述的方法Duceppe和Tabrizian(2009).总之,10毫克公顷被溶解在4毫升的缓冲区(0.1 M醋酸钠,0.1 M氯化钠,pH7.2)在磁搅拌.在HA是完全溶解,0.5毫克透明质酸酶(从牛睾丸、活动300 - 500单位/ mg,西格玛奥德里奇,美国)被添加到解决方案,然后保持24小时都在发抖孵化器在37摄氏度.酶分解反应后,这种混合物被过滤(Amicon超,MWCO 10负责,微孔公司,美国),然后是有机公顷的低分子量(MW < 10负责)和收集的centrifuged冻干(de laFuenteetal.,2008).这个resultingsubstance被溶解在蒸馏水(pH值为5.5).10毫克的CS是溶解在2%乙酸(pH5.5).两个HA和CS的解决方案是通过一个0.22米的过滤膜.CS的解决方案是搅拌的速度达到每天3000 rpm 30分钟,混合到HA解决方案和搅拌10分钟.八个不同的混合物还准备CS:哈重量比率:1:2,1:1,2:1,3:1,4:1,5:1,6:1,7:1.一定浓度的11.25 g /毫升公顷被用在所有的混合物,CS浓度各不相同:5.625,11.25,22.5,33.75,45岁,56.25,67.5,和78.25 g /毫升.所需的体积的25 g /毫升质粒DNA被轻轻地添加到CS / HA解决方案在温和移液形成复合物的所选N / P比率.N / P比被定义为的摩尔比积极的CS氨基组和消极的DNA磷酸基.这种混合物被vortexed迅速为3 - 5 s和左1 h在室温下配合完全形成.