Detection of usual and atypical aldehyde dehydrogenase alleles by mismatch amplification mutation assay

The genotypes of liver mitochondrial high-affinity aldehyde dehydrogenase-2 (ALDH2) are strongly associated with the drinking behavior and the alcohol liver diseases, since the individuals with atypical ALDH(2)(2) allele have higher levels of acetaldehyde in their plasma. The atypical ALDH(2)(2) allele has a nucleotide base transition (G-->A) in its exon 12. Based on this point mutation, we developed a rapid, reliable and inexpensive method, mismatch amplification mutation assay (MAMA), for the determination of human ALDH2 usual and atypical alleles. Two pairs of primers were designed for the amplification of the usual ALDH(2)(1) allele and the atypical ALDH(2)(2) allele, respectively. If the sample for the detection was heterozygous, it could be amplified by both of the primers. The product of polymerase chain reaction (PCR) of ALDH2 exon 12 could be easily screened by electrophoresis on a 2% agarose gel. The results of the MAMA method were further confirmed by sequencing. In the total of fifty samples from unrelated healthy Chinese Han people from Wuhan, China, the frequency of atypical ALDH(2)(2) allele was found to be 12%.

活性/可控聚合法合成旋光性聚甲基丙烯酸（上艹下孟）酯

本论文合成了旋光性甲基丙烯酸（上艹下孟）酯（MnMA）、并分别用阴离子聚合，基团转移聚合（GTP），自由基聚合及原子转移自由基聚合（ATRP）方法对其进行了聚合研究，考查了不同聚合方法对其聚合能力、聚合物微观结构及旋光性的影响，主要结论如下：1.在MnMA的阴离子聚合中，分别加人(+)DDB、（-）Sp, TMEDA等配体后，聚合能力明显增强。2.（-）一MAMA的阴离子聚合所得聚合物的比旋光值随其分子量的增加略有增加；手性与非手性引发体系及不同聚合方法对聚甲基丙烯酸莹醋的比旋光值影响均较小。2.不同聚合方法对聚甲基丙烯酸盏酷的立构规整性的影响程度不同，且所得聚合物的全同含量均较低。4.以甲基丙烯酸三苯基甲酷（TrMA）作为活性预聚物可以诱导MnMA、乙烯基三苯基磷(VTPP）等大位阻单体聚合，实现螺旋诱导不对称聚合。5.采用ATRP及反向ATRP对（-）-MnMA进行了聚合研究，结果表明：(1)（-）MriMA的非均相ATRP研究表明，不同引发剂及配体对其聚合影响程度不同，2一澳丙酸乙醋(2-EBP)/CuCl/2,2'-联毗陡(bipy)/THF体系的引发效率最高，体系所得聚合物的分子量分布最窄，可控性最好。(2)（-)MnMA的两均相ATRP体系2-BP/CuCl/(-)-Sp(or dHbpy)ffHF所得聚合物分子量随转化率的变化基本呈直线关系，分子量分布均控制在1.2以下。(3)对(-)MnMA反向ATRP研究表明，AIBN/CuC12/bipy(or (-)-Sp)fFBF体系的动力学曲线呈一级线性关系，分子量随转化率的变化基本呈直线增加，可控性较好。(4) ATRP及反向ATRP体系中所得聚合物的比旋光值绝对值随分子量的增加略有减小。