ARMS-PCR method for detecting multiple NPM1 mutations / 中国实验血液学杂志

This study was aimed to establish a simple, sensitive detection method for multiple NPM1 mutations, so as to reduce the omission ratio of NMP1 mutant detection. Recombinant plasmids containing wide-type NPM1 and the most common mutations (A, B, C, D) were constructed as the detection objects. The ARMS-PCR for detecting multiple NPM1 mutations was established through designing a pair of specific primers whose 3' end base matched with four mutants (A,B,C,D), but did not matched with wild type NPM1 according to the different base sequence of NPM1 mutants. The feasibility of the ARMS-PCR method was evaluated by assessing the detection range and the sensitivity and comparing with direct sequencing. The results showed that the recombinant plasmids were constructed successfully by restriction analysis and DNA sequencing. The four mutants but not wild type NPM1 were detected by using ARMS-PCR, the detection range of the method was 10(3) copies/ml -10(9) copies/ml and the sensitivity was 0.01%, while the direct sequencing method could not detect the mutations if mutation was less than 10%. It is concluded that the high sensitive ARMS-PCR is established for detecting the four mutations of NPM1 and more than 95% mutants can be detected by this method, providing a new detection method for clinical NPM1 gene mutant.

Relationship between dehydroepiandrosterone and arteriosclerosis in premenopausal and postmenopausal women / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the correlation between dehydroepiandrosterone and arteriosclerosis in premenopausal and postmenopausal women.</p><p><b>METHODS</b>Forty premenopausal and 40 postmenopausal women were examined for serum concentrations of dehydroepiandrosterone and intima-media thickness of the carotid artery, and the serum concentrations of lipids, estrogen, endothelin, and E-selectin were also measured.</p><p><b>RESULTS</b>Compared with premenopausal women, the mean intima-media thickness was increased but dehydroepiandrosterone and estrogen levels were decreased in postmenopausal women. A significant inverse correlation was detected between the intima-media thicknesses and dehydroepiandrosterone level. The postmenopausal women had decreased antioxidation and elevated low-density lipoprotein level.</p><p><b>CONCLUSION</b>Arteriosclerosis is more likely to occur in women with low dehydroepiandrosterone level which causes decreased antioxidation and elevation of blood lipid levels.</p>

Protective effects of estrogen on mitochondria in human umbilical vascular endothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effects of estrogen on the mitochondria in human umbilical vascular endothelial cells (HUVECs).</p><p><b>METHODS</b>HUVECs were exposed to H2O2 at 250 micromol/L for 4 h with or without pretreatment with 17-estradiol (E2) and ICI182780. Complex IV activity of the cells was measured with chromometry, and 2, 7-dichlorofluorescein diacetate (DCFH-DA) was used to determine intracellular reactive oxygen species (ROS). Intracellular adenosine triphosphate (ATP) level was quantified with a luciferin- and luciferase-based assay.</p><p><b>RESULTS</b>Compared to the blank control group, H2O2 caused a decrease in complex IV activity, intracellular ATP level, and the cell viability, but elevated intracellular ROS. E2 pretreatment of cells significantly attenuated these effects of H2O2 exposure. ICI182780 administered prior to E2 pretreatment antagonized the protective effects of E2 against H2O2 exposure.</p><p><b>CONCLUSION</b>E2 offers mitochondrial protective effects on HUVECs, which is mediated by the estrogen receptors.</p>