由重组腺相关病毒1／2载体携带的LacZ报告基因在脐血间质干细胞中的表达

目的:观察由重组腺相关病毒1/2载体携带的LacZ报告基因在体外培养的脐血间质干细胞中的表达情况。方法:实验于2005-10/2006-02在上海交通大学附属新华医院科研中心完成。①以孕龄近60d的杂种妊娠犬的脐带血作为实验用脐血间质干细胞的标本来源。重组腺相关病毒1/2载体-lacZ基因(北京本原正阳基因技术有限公司)。②无菌条件下采集妊娠犬脐带血,置于预装有肝素的离心管中,与Hanks液1∶1混合均匀,叠加于相对密度为1.077的淋巴细胞分离液上,梯度离心分离后进行培养和扩增。③取第4代脐血间质干细胞,经胰酶消化后吹打成单细胞悬液,以5×104/孔接种于24孔板内,随机数字表法分为转染组20孔、空白对照组4孔。转染组将重组腺相关病毒1/2载体-lacZ报告基因(1×1012v.g.mL-1)用不含血清的IMDM培养液作系列滴度稀释,分为5个滴度,即感染复数分别为每个细胞1×102,1×103,1×104,1×105,1×106v.g,各感染复数均设4孔;空白对照组未加入病毒,只加入相同体积的不含血清的IMDM培养液。④转染72h后采用X-gal化学染色法进行检测,成功转染上LacZ基因的脐血间质干细胞其胞浆内会因合成半乳糖苷酶而呈阳性蓝染。每组样本随机选取5个视野,相差显微镜下计数半乳糖苷酶阳性细胞数,取均值即为LacZ基因细胞转染率。结果:①脐血间质干细胞生长情况及LacZ报告基因表达的检测:转染组病毒基因转染后未再见到明显的细胞增殖,转染72h后大部分细胞表达LacZ基因并合成半乳糖苷酶,X-gal染色呈蓝色,长梭形;4～6周后细胞形态趋向老化,长梭形逐渐变为宽扁形;8周后细胞均被蓝染,颜色明显较转染72h时深,细胞形态由长梭形变为不规则,明显老化。空白对照组细胞反应均为阴性。②LacZ报告基因细胞转染率测定结果:转染72h后,感染复数为每个细胞1×102v.g时,仅有少数细胞蓝染呈阳性;感染复数为每个细胞1×103,1×104,1×105,1×106v.g时,转染率分别为(43±5)%,(82±4)%,(95±4)%,(97±3)%。结论:体外培养的脐血间质干细胞能高效转染重组腺相关病毒1/2载体-lacZ报告基因,在一定感染复数范围内,细胞转染率随着感染复数的增加而升高。提示脐血间质干细胞是重组腺相关病毒1/2载体-lacZ报告基因转染的适宜靶细胞。     

Navigation-guided oral and maxillofacial surgery (reconstructive and plastic, orthognathic, oncology)

Purpose: Advance in the field of compeer assisted surgery enables the surgical procedures to be less invasive and more accurate for the support of diagnosis imaging, pre-operative simulation and intraoperative navigation.     

结直肠癌淋巴管生成与淋巴结转移相关检测的临床研究进展

结直肠癌是人类常见的可经淋巴道转移的恶性肿瘤,其淋巴管生成与转移过程中牵涉了多种生物标志物表达水平的改变,检测这些相关生物标志物可为结直肠癌的诊断、治疗和预后情况提供重大参考价值．本文主要综述近年来与结直肠癌淋巴管生成与淋巴结转移相关检测的临床研究进展．     

褶合曲线分析法测定复方茶碱片中六种主要成分

应用柱分配色谱,将复方茶碱片分离出各含三个组分的两组混合物,然后分别应用褶合曲线分析法,同时定量测定了两组混合物。六个被测成分的平均百分回收率和相对标准差分别为:咖啡因101.6,1.46%;非那西丁99.7,0.10%;苯巴比妥100.9,1.31%;可可碱99.9,0.81%;茶碱100.2,0.81%;氨基比林100.8,0.48%。     

Reliability of the Otago photoscreener A study of a thousand cases

The Otago photoscreener is a 35 mm single lens reflex camera in which the flash light comes from a narrow ring around the outer margin of its lens. The margin is also the limiting aperture of the optic system and in the centre of the lens is a flickering fixation light. In a colour photograph taken at a distance of 66 cm from the face of the subject who is accurately focusing on and fixing the camera fixation light with both eyes, the fundus reflex in each pupil is very dark red and the corneal light reflexes are symmetrical. If either or both eyes are not appropriately focused or fixing, the fundus reflex is brighter and yellow or white. This article describes a prospective trial of the performance of the Otago photoscreener in a series of 1000 infants with actual or suspected amblyopia, refractive error or strabismus. In this study photoscreening showed a sensitivity of 94% and a specificity of 79%. Photoscreening also identified some cases of esotropia and of refractive error which were missed on clinical examination. In this series photoscreening passed as normal three children with mild to moderately severe amblyopia. This represents les than 1 % of the clinically abnormal children.     

Neuronal migrational disorders: diffuse cortical dysplasia or the "double cortex" syndrome

We report a 10-year-old girl who presented with a single episode of partial complex seizure. Neurological examination and psychological evaluation were normal. Electroencephalogram showed abnormalities in the left temporo-occipital region. Magnetic resonance imaging revealed a bilateral layer of tissue iso-intense with gray matter, between the wall of the lateral ventricle and the cortex, extending from the frontal to the occipital region, and enlarged left lateral ventricle. The images indicated diffuse cortical dysplasia, also known as band heterotopia or "double cortex", and left cortical hemiatrophy.