Down-regulation of Notch-1 gene expression inhibits growth of TJ-905 glioblastoma

<正>Objective To study the inhibitory effect of siRNA on glioblastoma (GBM) Notch-1 gene expression in addition to the growth of TJ-905 glioblastoma. Methods Three small interference RNAs (siRNAs) targeting Notch1 gene,named siRNA1,siRNA2,siRNA3,synthesized chemically in vitro with gene bank BLAST. TJ-905 cells were transfected twice with the siRNA by using Oligofectamine 2000     

特发性肥厚性硬脑膜炎二例并文献复习

特发性肥厚性硬脑膜炎( idiopathic hypertrophic cranial pachymeningitis,IHCP)是一种罕见的以局灶性或弥漫性硬脑膜增厚为特点的炎性增生性疾病.     

原发性蝶骨大翼发育不良一例报道及文献复习

蝶骨大翼发育不良(sphenoid wing dysplasia,SWD)常见于神经纤维瘤病1型(neurofibromatosis type Ⅰ,NF-1),其在NF-1中的发病率约在3％～7％之间[1],原发性SWD极为罕见.现将收集的原发性SWD 1例患者资料并结合文献复习报道如下. 患者 男性,40岁,因"车祸后头痛、头晕2h"于2012年4月1日人青岛大学医学院附属医院治疗.体格检查可见左侧眼球外突、下移,未见明显搏动性突眼、皮肤色素沉着、咖啡牛奶斑和多发性神经纤维瘤.     

下调Notch-1基因表达抑制胶质母细胞瘤的增殖活性研究

目的 应用siRNA下调胶质母细胞瘤TJ-905细胞株Notch-1基因抑制肿瘤增殖活性.方法 实验设以Notch-1为基因靶点的siRNA1、siRNA2、siRNA3转染组、阴性对照组和空白对照组,采用OligofectamineTM2000二次转染方法将 siRNA转移到细胞内.实时荧光定量PCR检测siRNA对Notch-1基因表达的抑制作用,筛选出最优siRNA序列用于后续研究;四唑盐(MTT)比色法检测细胞增殖活性;使用Notch-1 siRNA或阴性siRNA转染的TJ-905细胞悬液注射裸鼠腋下,观察移植瘤生长情况,比较肿瘤体积,观察3周后处死动物,切片行HE染色鉴定.结果 (Ⅰ)siRNA明显抑制Notch-1 mRNA的表达水平,空白对照组、阴性对照组、siRNA1、2、3转染组Notch-1 mRNA 表达率分别为1.00±0.07、1.04±0.05、0.11±0.02、0.12±0.01、0.77±0.03,siRNA1为最优序列.(2)siRNA1转染组细胞增殖活性显著降低.(3)接种Notch-1 siRNA1转染的TJ-905细胞裸鼠肿瘤生长速度明显低于接种阴性转染的TJ-905细胞裸鼠,裸鼠处死后观察肿瘤体积:Notch-1 siRNA1转染组为(748.3±154.3)mm3,阴性转染组为(1739.2±249.7)mm3,病理切片HE染色证明为多形性胶质母细胞瘤.结论 化学合成的靶向Notch-1基因的siRNA可以显著地下调Notch-1基因表达水平,抑制肿瘤增殖活性.为进一步进行胶质瘤的基因治疗研究奠定基础.

Abstract：

Objective To study the inhibitory effect of siRNA on glioblastoma (GBM) Notch-1 gene expression in addition to the growth of TJ- 905 glioblastoma.Methods Three Small interference RNAs (siRNAs) targeting Notch- 1 gene named siRNA1,siRNA2,siRNA3 were synthesized chemically in vitro with gene bank BLAST.TJ-905 cells were transfectedtwice with the siRNA by using OligofectamineTM2000.The nontransfected cells and nonspecific siRNAs transfected cells were taken as blank and nonsense controls.Down - regulation of Notch- 1 was demonstrated by real- time RT- PCR,according to the result of QRT- PCR we therefore selected the most effective siRNA for further study.Cell proliferation was measured by MTT analysis.Male Balb/C nude mice were injected subcutaneously with Notch- 1 siRNA- or nonsense siRNA- transfected TJ-905 cells,tumor sizes were measured every 4 days.HE stain was used to determine the property of tumor.Resuts(1) The expression of Notch- 1 mRNA in blank,nonsense controls,siRNA1,2,3 groups were 1.00±0.07,1.04±0.05,0.11 ±0.02,0.12 ±0.01,0.77 ±0.03 by real-time RT- PCR scan analysis,the most effective siRNA was siRNA1.(2) An inhibitory proliferation and growth can be induced in siRNA1 transfected group.(3)Nude mice xenografted with cell suspensions from the nonsense siRNA group developed tumors with a significantly increased volume (from the 18th days) as compared to mice that received the Notch- 1 siRNA1- treated cells.The final tumor volume were less in nude mice injected with Notch- 1 siRNA (748.3 ±154.3 )mm3 compared to nonsense si RNA injection (1739.2 ± 249.7 )mm3,HE stain demonstrate that tumor was multiformity glioblastoma.Conclusion The chemically synthesized siRNA targeted Notch- 1 gene could down -regulate the expression of Notch- 1.In addition,an inhibitory proliferation and growth were induced when compared with the control cells in vitro and in vivo.It was suggested that the suppression of Notch- 1 expression and the inhibition of growth provide a new way to glioma gene therapy.     

医生的助手——移动医疗App

自2012年移动医疗应用开始兴起后,医疗类App就逐步成为医生常用的应用工具。在国内,目前有超过100款的医生专用App,它们最大的特点是帮助医生提高工作效率,其功能主要集中在医学信息交流、医患交流、患者管理等方面。丁香客和用药助手是丁香园旗下面向医生群体的医疗APP。丁香客将医生群体通过关系网络凝聚在一起,帮助业内人士建立更广泛的学术圈子,扩大学术影响力。     

远离噪声,安全用耳

3月3日是国际爱耳日,今年的主题是"make listening safe",即"安全用耳,保护听力"。来自世界卫生组织的报告显示:全球有3.6亿人有听力残疾,占全球总人口的5.3%;其中,我国有听力残疾的达2780万。耳鼻喉科专家指出,儿童及老年人是听力障碍的多发人群,所有耳聋都必须以预防为主,一旦发生,就不可能恢复到原先的听力水平。世界卫生组织近日发布的研究报告称,全球约有11亿青少年正在受到个人视听设备和娱乐场所     

锁骨骨折86例治疗体会

我队于1993～2003年期间共收治锁骨骨折86例，现将治疗情况及疗效报告如下：     

卫生员培训中存在的问题与对策探讨

卫生员是部队基层卫生工作的一支重要力量，无论是在平时卫生防病，还是战时火线抢救中都发挥着重要作用。卫生员训练队作为部队培养卫生员的基地，其培训质量的高低直接影响着卫勤保障能力。     

靶向Notch-1基因的siRNA对人脑胶质母细胞瘤细胞裸鼠体内成瘤性的影响

背景与目的:近些年的研究发现,Notch-1基因在人脑胶质母细胞瘤的发生、发展中起到了重要的作用.本研究旨在观察靶向Notch-1基因的小干扰RNA(siRNA)对人脑胶质母细胞瘤裸鼠体内成瘤性的影响.方法:以OligofectamineTM为载体将靶向Notch-1基因的SiRNA转染人脑胶质母细胞瘤细胞系TJ905中,并设立无意序列siRNA(nonsense siRNA)转染组.然后,把经Notch-1 siRNA和经无意序列siRNA转染的细胞及未经转染的细胞分为3组.每组细胞悬液注射裸鼠5只,注射量按1×107/mL无血清培养基注射入每只裸鼠皮下200 μL,观察动物的成瘤情况,每隔3 d记录-下裸鼠移植瘤的体积V=(ab2)/2(a为肿瘤的长径,b为肿瘤的宽径),并在接种成瘤后对各组动物进行相应的SiRNA/Oligofectamine TM混合物的瘤内注射治疗,对照组注射等量的PBS.观察20 d后处死动物,取出肿瘤组织,常规石蜡包埋,组织切片HE染色并进行Notch-1的免疫组化染色测定.结果:裸鼠处死后观察肿瘤直径:Notch-1 siRNA转染组为(1 203±206)mm3,siRNA转染组为(2 241±401)mm3,对照组为(2 309±466)mm3,Notch-1 siRNA转染组与无意siRNA转染组,以及Notch-1siRNA转染组与对照组之间比较均有统计学意义(P<0.05).免疫组化结果显示:Notch-1的表达在Notch-1siRNA转染组与其他两组之间的比较也同样具有统计学意义(P<0.01).结论:靶向Notch-1基因的siRNA可以通过有效地降低肿瘤细胞Notch-1的表达来抑制裸鼠皮下胶质母细胞瘤移植瘤的生长,但还需要对siRNA的合理使用剂量与转染方式进行更加深入的研究.     

凉血止血方佐治ATG治疗无效及复发的重型再生障碍性贫血40例

重型再生障碍性贫血(SAA)是免疫介导的,以骨髓造血干细胞及造血微环境损伤、骨髓脂肪化和外周血全血细胞减少为特征的造血功能衰竭综合征。临床上表现为严重的感染、出血和贫血。抗胸腺细胞球蛋白(ATG)等免疫抑制治疗是目前治疗SAA的主要措施,但临床上有10%～30%患者