Analysis of glioma cell lines for amplification and overexpression of MDM2

Recently, amplification of the gene encoding a p53 binding protein, MDM2, was determined in 8% of the cases constituting a large series of glioblastomas. Here we have utilized Southern blot analysis to examine 30 cell lines established from such tumors, and our investigation has revealed large increases in MDM2 gene dosage in two cases, one of which showed coamplification of the CDK4 gene that resides in close proximity to MDM2 in chromosomal region 12q13–14. Northern analysis demonstrated overexpression of MDM2 mRNA in the two cell lines with gene amplification, and overexpression of MDM2 protein was evident in each of these by immunohistochemical and Western blot analysis. Analysis of TP53 cDNAs revealed normal TP53 sequences in the cell lines with MDM2 amplification; these results are consistent with those of previous studies suggesting that MDM2 amplification occurs only in tumors expressing wild-type p53. In total, these data suggest that MDM2 amplification in glioblastoma cell lines occurs at a frequency (6.7%) comparable to that determined in primary tumors; occurs in cell lines expressing wild-type p53; and can involve the coamplification of additional genes.     

掌背动脉皮瓣的应用解剖

目的：手指软组织损伤，使用手背动脉皮瓣进行修复，手术简单，转移方便，并能很好满足手指修复中对于弹性，厚度，韧度，感觉和活动功能等诸多要求，方法：４３只上肢标本，色素动脉灌注，解剖掌背动脉及分支，结果：观测掌背动脉起源，形态，分支吻合，管径，并计算各吻合点的自比例定位，结论；提出掌背动脉轴形皮瓣应用范围及可行性，为临床提供解剖学资料和手术操作要点。     

Formation of quinone methide in the self-cure of quinoid phenol formaldehyde resin

The study of the reaction of some methylenebisphenols with chloranil in ethanol and also of a phenol formaldehyde condensate prepared by acid catalysis, reveals that quinone methide is formed in the self-cure of quinoid phenol formaldehyde resin. The quinone methide reacts with ethanol to give an adduct. The structure of the alcohol adduct is identified by NMR and IR data and by means of pyrolysis.     

一种用ICA去除瞬态诱发耳声发射伪迹的新方法

如何去除伪迹是瞬态诱发耳声发射检测中一个关键的问题。本研究提出了一种用ICA去除伪迹的新方法。首先用四组线性增长的刺激声在耳道内录音 ,得到的波形是瞬态诱发耳声发射和伪迹的混叠。因为伪迹和瞬态诱发耳声发射是统计独立的 ,而且伪迹随刺激声的变化线性增长 ,而瞬态诱发耳声发射随刺激声的变化非线性增长 ,逐渐趋于饱和 ,所以它们在混叠信号中具有不同的混叠系数。用ICA算法可以将各独立分量及混叠矩阵估计出来 ,伪迹是其中的一个独立分量。然后将伪迹的波形置零后再进行一次混叠 ,便达到了去除伪迹的目的。最后通过与传统的DNLR方法比较 ,证明这种方法是有效的     

Wg3h细胞系转染PSV_2-neo质粒前后生长特性及表面超微结构改变的研究

本文对转染PSV_2-neo质粒后的Wg3h细胞系(Wg3h-neo)在长期传代中的生长特性和表面超微结构与母系Wg3h细胞进行了比较研究。结果发现:转染后Wg3h细胞的DNA合成及生长速度明显高于母系Wg3h细胞,生长饱和密度增大。在软琼脂培养中不形成集落,接种在裸鼠不长肿瘤。在扫描电镜下,细胞表面的微绒毛较母系Wg3h细胞丰富。Southern印迹杂交实验证明PSV_2-neo质粒已整合到宿主细胞的基因组中。转染后Wg3h细胞的生长特性和表面超微结构均发生了某些转化特征。     

Fos oncoprotein expression in the rat forebrain following muscimol-induced absence seizures

Fos oncoprotein expression is a marker of neuronal activation following seizures. Here, using this method we examined the anatomical locations of muscimol-induced absence seizures in the rat forebrain. Six hours after a systemic injection of muscimol a massive Fos immunoreactivity appeared in the olfactory system, retrosplenial cortex and paraventricular thalamic nucleus, whereas other cortical areas contained low level of Fos expression. These results provide the first functional morphological evidence suggesting that these forebrain structures with Fos expression may play an important role in the pathophysiology of muscimol-induced absence seizures.     

Digital detection of genetic mutations using SPC-sequencing

Deletion or insertion mutations lead to a frameshift that causes misalignment between wild-type and mutated allele sequences, making it difficult to identify such mutations unambiguously by using electrophoresis-based DNA sequencing. We have previously established the feasibility of an accurate DNA sequencing method using solid-phase capturable (SPC) dideoxynucleotides and MALDI-TOF mass spectrometry on synthetic templates, an approach we refer to as SPC-sequencing. Here, we report the application of SPC-sequencing in characterizing frameshift mutations by using the detection of the BRCA1 gene mutations 185delAG and 5382insC as examples. In this method, Sanger DNA sequencing fragments are generated in one tube by using biotinylated dideoxynucleotides. The sequencing fragments carrying a biotin moiety at the 3' end are captured on a streptavidin-coated solid phase to eliminate excess primer, primer dimers, and false stops. Only correctly terminated DNA fragments are captured, subsequently released, and analyzed by mass spectrometry to obtain digital DNA sequencing data. This method produces distinct doublet mass peaks at each point in the mass spectrum beyond the mutation site, facilitating the accurate characterization of the mutation. We have compared SPC-sequencing with electrophoresis-based sequencing in characterizing the above BRCA1 mutations, demonstrating the significant advantage offered by SPC-sequencing for the accurate identification of frameshift mutations.