MSI2通过下调circRNA_001214促进急性髓系白血病细胞HL60生长

Musashi-2（MSI2）是一种RNA结合蛋白质,对维持造血干细胞功能具有重要作用。研究表明,MSI2高表达能促进急性髓系白血病（acute myelocytic leukemia, AML）进展,但其作用机制尚不明确。本研究稳定沉默HL60细胞MSI2后,第1、2、3、4 d对照组的相对细胞生长率分别为1.931 ± 0.027、3.070 ± 0.073、4.017 ± 0.092和4.215 ± 0.246;敲减组分别为1.927 ± 0.035、2.564 ± 0.090、2.825 ± 0.097和3.223 ± 0.182,两组相比具有统计学差异,P<0.001;细胞凋亡明显增加(7.967% ± 0.698% vs 3.400% ± 0.322%., P<0.01);G₀/G₁期细胞比例明显增高（67.430% ± 4.390% vs. 50.360% ± 2.160%, P<0.01）;NUMB蛋白明显上调,LEF1明显下降。环状RNA（circular RNA, circRNA）芯片筛选和荧光定量PCR验证显示,MSI2沉默组circRNA_001214表达水平是对照组3.48倍。这一结果也在NALM6细胞得到证实。进一步用生物信息学分析,显示circRNA_001214最可能与miR-1273a、miR-1273e和miR 5095结合,进而影响参与细胞凋亡相关基因（CYCS、AKT1、BAX、TNFRSF10A、TNFRSF10D）、Wnt信号基因（WNT4、WNT2B、WNT7B、 DKK2、SFRP1、CSNKE1和LEF1）以及参与细胞代谢相关基因（RPE, PGAM4, PGAM1, TAT, CBS、RPE、SUCLG2、PGAM4、PGAM1和 IDNK）。总而言之,MSI2可能通过干扰circRNA_001214生成,减少靶miRNA对凋亡、Wnt信号及细胞代谢相关基因表达的影响,促进细胞生长。     

Free radical damage to cultured porcine aortic endothelial cells and lung fibroblasts: Modulation by culture conditions

Summary Culture conditions modulating cell damage from xanthine plus xanthine oxidase-derived partially reduced oxygen species were studied. Porcine thoracic aorta endothelial cells and porcine lung fibroblasts were maintained in monolayer culture. Cells were prelabeled with⁵¹Cr before xanthine plus xanthine oxidase exposure. Endothelial cells showed 30 to 100% more lysis than fibroblasts and thus seemed more sensitive to this oxidant stress. The effect of cell culture age, as indicated by population doubling level (PDL), was examined. Response of low PDL endothelial cells and fibroblasts subjected to oxidant stress was compared with the response of PDL 15 cells. Both low PDL endothelial cells and fibroblasts responded differently to the lytic effect of xanthine oxidase-derived free radicals than did higher PDL cells. Specific activities of the antioxidant enzymes catalase, managanese superoxide dismutase, copper-zinc superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase were measured in both low and high PDL fibroblasts and endothelial cells. Antioxidant enzyme specific activities could only partially explain the differences in response to oxidant stress between fibroblasts and endothelial cells and between low and high PDL cells. Cell culture medium composition modulated the rate of production, and relative proportions of xanthine plus xanthine oxidase-derived partially reduced species of oxygen, i.e. superoxide, hydrogen peroxide, and hydroxyl radical. Serum content of medium was important in modulating free radical generation; superoxide production rates decreased 32%, H₂O₂ became undetectable, and hydroxyl radical generation decreased 54% in the presence of 10% serum. The medium protein and iron content also modulated free radical generation. The data suggest that cell culture media constituents, cell type, and cell culture age greatly affect in vitro response of cells subjected to oxidant stress. Research supported by American Lung Association Fellowship Training Grant and Research Training Grant, the R. J. Reynolds Corporation, and National Institutes of Health Grants HL29784 and 1 HL 23805.     

Transition Metals Potentiate Paraquat Toxicity

The involvement of transition metal ions in paraquat toxicity was studied in bacterial model system. We show that the addition of micromolar, or lower, concentrations of copper dramatically enhanced the rate of bacterial inactivation. In contrast, the addition of chelating agents totally eliminated the killing of E. coli. No inactivation was observed under anaerobic exposure to paraquat, both in the absence and presence of copper. However, in the presence of copper, the anaerobic addition of hydrogen peroxide resulted in complete restoration of inactivation as under aerobiosis.

Paraquat either produces superoxide ions or directly reduces bound copper ions in a catalytic mode. The reduced cuprous complexes react with hydrogen peroxide to locally form hydroxyl radicals (OH) which are probably responsible for the deleterious effects.

This study indicates the involvement of a site-specific metal-mediated Haber-Weiss mechanism in paraquat toxicity. It is in agreement with earlier observations that copper unusually enhance biological damage induced by either superoxide or ascorbate.     

The influence of mineral salts on fecundity of the water flea (Daphnia magna) and the implications on toxicity testing of industrial wastewater

The effects of mineral salts constituting water hardness on fecundity ofDaphnia magna were assessed. Of the salts tested, increased concentrations of NaHCO₃ and MgSO₄ had no effect on fecundity, CaSO₄ significantly increased fecundity, and KCl significantly reduced fecundity. The number of offspring produced per daphnid was correlative to the CaSO₄ concentration at CaSO₄ concentrations between 91 and 2100 mg/ℓ. The effects of CaSO₄ on daphnid fecundity could influence the interpretive outcome of industrial wastewater toxicity tests using this species when the waste and dilution waters contain different concentrations of CaSO₄. It is recommended that when performing these tests, dilution water be sampled at the intake site of the industry's water source, thus assuring initial comparability of the waste and dilution waters. The CaSO₄ content of the water prior to and after industrial use should be determined to identify any alterations of CaSO₄ concentration during use. Identification of CaSO₄ concentration differences can aid in the interpretation of effects associated with the wastewater.     

Factors to consider in performing survival studies with insect cells

Summary Insect cell lines are not well-suited to colony formation in liquid medium following low-density cell plating. The present studies demonstrate that the time of addition of fetal bovine serum to the culture medium and the number of γ-irradiated feeder cells added to each plate are important factors in developing a useful colony formation assay. TN-368 lepidopteran and WR69-DM-1 dipteran cell lines were used for these experiments. Both cell types display increased plating efficiencies if serum is added to the medium one or more days prior to plating as compared to adding serum immediately before plating. Growth curves obtained by seeding cells at higher densities also indicate that cell growth is slightly better if serum is added one or more days before seeding. These findings are especially important for survival and toxicity studies because the results demonstrate that even seemingly minor factors involved in cell survival assays may benefit treated cells to a greater degree than untreated control cells, thus providing an erroneous assessment of cell survival. This work was supported by USPHS grant R01-CA34158, awarded by the National Cancer Institute, DHHS, Bethesda, MD.     

Effects of CuDIPS and tween 80 on SJL/J tumorigenesis

Plasma copper and zinc levels were measured in SJL/J mice, an inbred strain characterized by a high, spontaneous incidence of reticulum cell sarcoma (RCS). The changes with age in mean concentrations of these metals were consistent with a physiological response that is required for remission of neoplasia. Treatment of SJL/J mice with a copper complex, Cu(II)(3,4-diisopropylsalicylate)₂ (Cu 3,5-DIPS), dissolved in a 10% Tween 80-saline solution revealed a decrease in survival and decline in the incidence of RCS at 52 wk of age. The toxic effects of Cu 3,5-DIPS therapy appeared to be related to the intraperitoneal route of administration and to extracellular deposition of collagen. The inhibitory effect on tumor development was not related to Cu 3,5-DIPS. Rather, Tween 80 was found to be the factor of importance.     

Chemical defence in chrysomelid eggs and neonate larvae

ABSTRACT. Eggs and neonate larvae of chrysomelid beetles (sub-tribes Chrysomelina and Phyllodectina) were investigated for the presence of defensive substances.
The two isoxazolinone glucosides (compounds 1 and 2), characteristic of the adult defence secretion, were detected in the eggs of all studied species. Compound 2, containing a nitropropionate, is always present in concentrations (above 10^-2 M), which are highly deterrent to the ant Myrmica rubra. This compound is not at all or only slightly toxic to ants at 10^-2 M. Compound 1, devoid of nitropropionate, is a minor constituent, and is neither deterrent nor toxic to ants.
The five Chrysomela species studied and Phratora vitellinae also sequester salicin in their eggs in amounts highly deterrent and toxic to ants. A single Chrysomela egg often contains enough salicin to kill an ant. While the isoxazolinones are discarded with the egg shells, salicin is used by neonate larvae as a precursor for the production of salicylaldehyde in the thoracic defence glands, already functional at hatching. No salicin could be detected in the eggs of those species whose larvae produce cyclopentanoid monoterpenes, even if they feed on Salicaceae. No larva of any species seems to be able to produce detectable amounts of monoterpenes at birth. A very early defence, possible only in those species using salicin as the precursor for their defensive secretion, could be highly advantageous in protecting the clustered larvae during the long process of hatching and in avoiding cannibalism between siblings.
Only trace amounts of oleic acid were found in the eggs of Gastrophysa viridula , in contrast to previous reports on its presence in large quantities in the American G. cyanea.     

The in vitro screening of methylated 4-oxypiperidine compounds for inhibition of protein synthesis in a rabbit reticulocyte cell-free translation system

A variety of methylated 4-oxypiperidine derivatives were tested for their ability to inhibit protein synthesis in vitro. A direct correlation was found between the extent of methylation of these compounds and their inhibitory activity in a rabbit reticulocyte lysate cell-free translation system.Abbreviation IC₅₀ 50% inhibitory concentration     

Chronic toxicity of three insecticides (carbaryl,fenthion and lindane) in the freshwater snail Lymnaea stagnalis

A chronic intoxication with carbaryl, fenthion and lindane was induced in young snails. The parameter k of the von Bertalanffy's equation showed clearly the growth changes induced by these insecticides. In all cases the fecundity of intoxicated snails was reduced. Among these three insecticides, lindane was the most toxic, carbaryl the least.