HSV-TK/GCV自杀基因系统治疗宫颈癌的研究

目的探讨HSV-TK/GCV自杀基因系统对人宫颈癌细胞系Hela体外及体内杀伤作用及其产生的旁观者效应.方法采用脂质体转染法将GINaTK载体转入包装细胞PA317.取病毒上清液感染人宫颈癌细胞Hela,得到带有HSV-TK基因的Hela/TK细胞,并将其分别用于体外和体内实验.结果载体HSV-TK导入了PA317细胞.体外实验结果显示,当Hela/TK细胞数占混合细胞10%时,低浓度(10μg/ml)的GCV就可将50%左右的肿瘤细胞杀死.体内实验结果显示GCV可明显抑制Hela/TK细胞在BALB/C小鼠体内的肿瘤形成.经GCV治疗后,肿瘤体积分别较对照组肿瘤体积缩小约11.1%、30.6%和47.2%(均P＜0.001);RT-PCR检测HSV-TK基因在肿瘤组织中有表达;实验组肿瘤组织与对照组相比存在明显的病理学改变.结论逆转录病毒可介导HSV-TK基因转入人宫颈癌细胞Hela并获稳定表达,HSV-TK/GCV自杀基因系统在体内外对宫颈癌细胞均有杀伤作用,且存在明显的旁观者效应.     

肝癌特异性超抗原SEA(D227A)基因表达载体的构建与表达

目的：构建受AFP顺式作用元件调控的超抗原表达载体，将SEA(D227A)特异性的表达于AFP阳性肝癌细胞膜表面。方法：PCR扩增AFP基因启动子、增强子、linker—CD80tm和SEA(D227A)。将上述片断插入逆转录病毒载体pLXSN的多克隆位点，构建AFP基因顺式作用元件调控的肝癌特异性减毒超抗原表达载体(pLXSN SEA(D227A)—linker—CD80tm)。通过脂质体介导，以表达载体转染表达或不表达AFP的肿瘤细胞系，用RT—PCR和间接免疫荧光染色，检测SEA的表达。结果：成功地将AFP基因的启动子、增强子、linker—CD80tm和SEA(D227A)克隆到逆转录病毒载体pLXSN的多克隆位点，酶切鉴定和DNA序列分析无误，RT—PCR和间接免疫荧光法检测证实，SEA(D227A)能在AFP阳性的肝癌细胞膜特异性表达。结论：AFP顺式作用元件修饰的超抗原表达载体的构建，为下一步用其强化肝癌的免疫治疗奠定了基础。     

携带增强绿色荧光蛋白基因的Id2真核表达载体构建

目的：构建大鼠Id2基因真核荧光表达载体，为骨骼肌的组织工程研究提供有效的分子工具。方法：利用RT-PCR的方法扩增出Id2全长cDNA，利用T4 DNA连接酶将载体pGEM-T和Id2 cDNA进行连接，构建克隆载体，经限制性内切酶EcoR1酶切pGEM-Id2克隆载体和pEGF-C2真核表达载体，构建出重组真核表达载体pEGFP-C2-Id2，经酶切分析、PCR鉴定及DNA测序证实cDNA片段大小和序列的正确性；并通过细胞转染技术将Id2基因导入L6成肌细胞中。结果：经酶切分析和序列测定证实pEGFP-C2-Id2含大小正确的正向Id2 cDNA片段，获取了转染外源性Id2基因的L6细胞。结论：我们成功构建了同时携带有G418筛选位点和增强绿色荧光蛋白的Id2真核表达载体？     

Comparing Principal Components Analyses of Evoked Potentials Recorded from Heterogeneous Groups of Subjects

Principal components analysis of evoked potentials differing between groups presents an interpretive problem, particularly in psychiatric research. Two sets of principal components and associated factor scores may appear to differ. The issue is to determine the extent to which visually differing principal components and resultant factor scores span the same factor space. Sets of evoked potentials from controls and from schizophrenics were each subjected to principal components analysis, from which factor score coefficients were computed for all subjects. This allowed determination of the extent to which (1) the two sets of basis waves were similar and (2) the factor scores resulting from the set of basis waves derived from principal components analysis of the control subjects' evoked potential data adequately represented those of the schizophrenics and vice-versa. Canonical correlation analyses indicated substantial similarities between the principal component structures (sets of basis waves). Multiple correlational analyses confirmed that the basis waves from either group spanned the other group's factor space. Factor scores from either set of basis waves were highly correlated. These results suggest that principal component structures derived from evoked potentials of a control group may be used in computing evoked potential factor scores of psychiatrically diverse populations even though the average evoked potentials of the groups may differ in several ways.     

结核分枝杆菌Ag85B与IL-12基因真核共表达质粒的构建及表达

目的 :构建结核分枝杆菌Ag85B和鼠IL 12基因的共表达载体pBud85B IL12。方法 :将结核分枝杆菌Ag85B基因和鼠IL 12基因同时克隆入含多启动子的共表达载体pBudCE4 .1中 ,构建真核共表达质粒pBud85B IL12。以pBud85B IL12转染COS 7细胞 ,通过RT PCR及ELISA方法检测目的基因的表达。结果 :在COS 7细胞中同时可检测到Ag85B和IL12的表达。结论 :pBud85B IL12共表达质粒的成功构建 ,为对其免疫原性、免疫反应性及免疫保护作用的进一步研究奠定了基础     

基于主成分分析-神经网络的非编码RNA预测

预测非编码RNA对认识其调控功能具有重要意义。选择单核苷酸和二核苷酸出现频率作为神经网络分类特征,运用主成分分析方法降低输入数据的维数,去除数据间的相关性,采用Levenberg-Marquardt算法改善网络训练速度。对数据集的测试结果表明,此方法对细菌混合ncRNA的预测精度达到81.3%,对原核生物tRNA的预测精度达到93.3%,表明该方法能有效预测原核生物ncRNA。预测结果还发现两种古细菌的ORF序列在序列特征上与其它细菌和古细菌存在差别。     

Responses of calves to acute stress: individual consistency and relations between behavioral and physiological measures

The present study examined the consistency over time of individual differences in behavioral and physiological responsiveness of calves to intuitively alarming test situations as well as the relationships between behavioral and physiological measures. Twenty Holstein Friesian heifer calves were individually subjected to the same series of two behavioral and two hypothalamo-pituitary-adrenocortical (HPA) axis reactivity tests at 3, 13 and 26 weeks of age. Novel environment (open field, OF) and novel object (NO) tests involved measurement of behavioral, plasma cortisol and heart rate responses. Plasma ACTH and/or cortisol response profiles were determined after administration of exogenous CRH and ACTH, respectively, in the HPA axis reactivity tests. Principal component analysis (PCA) was used to condense correlated measures within ages into principal components reflecting independent dimensions underlying the calves' reactivity. Cortisol responses to the OF and NO tests were positively associated with the latency to contact and negatively related to the time spent in contact with the NO. Individual differences in scores of a principal component summarizing this pattern of inter-correlations, as well as differences in separate measures of adrenocortical and behavioral reactivity in the OF and NO tests proved highly consistent over time. The cardiac response to confinement in a start box prior to the OF test was positively associated with the cortisol responses to the OF and NO tests at 26 weeks of age. HPA axis reactivity to ACTH or CRH was unrelated to adrenocortical and behavioral responses to novelty. These findings strongly suggest that the responsiveness of calves was mediated by stable individual characteristics. Correlated adrenocortical and behavioral responses to novelty may reflect underlying fearfulness, defining the individual's susceptibility to the elicitation of fear. Other independent characteristics mediating reactivity may include activity or coping style (related to locomotion) and underlying sociality (associated with vocalization).     

应变率成像在胎儿房性心律失常诊断中应用的初步探讨

目的探讨应用速度向量成像（VVI）技术对胎儿房性心律失常时心肌结构力学及形变学影响的应变与应变率研究，评价胎儿心律失常时对心肌应激状态、运动传导就及功能的影像。方法对检出的77例胎儿心律失常（其中室上性心动过速（SVT）12例、65例为频发性房性早搏（PAC））进行VVI成像技术分析，检测心律失常时胎儿心肌运动速度向量变化及对应变与应变率影响，对照组80例心率正常胎儿。结果胎儿SVT12例中7例持续发作48h者治疗前心室率250—310bpm，均合并心衰及出现水肿，VVI速度向量振幅明显降低，其应变与应变率明显低于对照组（P〈0．01）；阵发一陛SVT5例VVI速度向量振幅-过性降低，心肌应变与应变率无变化；频发PAC组示早搏时速度向量振幅改变，且运动方向同步，心肌运动三维数据定量可显示旱搏及代偿间期的三维模式，应变与应变率与对照组无差异（P〉0．05）。结论VVI成像通过心律失常心肌结构力学变化解析胎儿心律失常，特别是对胎儿心动过速时的快速心率可进行实时动态分析。及应用M型扫描法确认心动周期使胎儿心肌应变与应变率分析成为可行。     

Frequency domain models of the EEG

The structure of the normal resting EEG crosspectrum SVV(omega) is analyzed using complex multivariate statistics. Exploratory data analysis with Principal Component Analysis (PCA) is followed by hypothesis testing and computer simulations related to possible neural generators. The SVV(omega) of 211 normal individuals (ages 5 to 97) may be decomposed into two types of processes: the xi process with spatial isotropicity reflecting diffuse, correlated cortical generators with radial symmetry, and processes that seem to be generated by more spatially concentrated, correlated sources. The latter are reflected as spectral peaks such as the process. The eigenvectors of the xi process are the Spherical Harmonic Functions which explains the recurring pattern of maps characteristic of the spatial PCA of qEEG data. A new method for estimating sources in the frequency domain which fits dipoles to the whole crosspectrum is applied to explain the characteristics of the localized sources.