Metabolic profiling of umbilical cord blood in macrosomia.

BACKGROUND/OBJECTIVE: The term macrosomia is used to describe neonates with a birth weight of 4000 g or more. Macrosomia is a potential risk factor for obesity and metabolic syndromes in postnatal and adult life, yet little is known about its associations with metabolic difference in the early age. We performed metabolic profiling of umbilical cord blood to discover differential metabolites of macrosomia. METHODS: We conducted a case-control study of full-term singletons with normal maternal glucose tolerance [50 cases (macrosomia, birth weight ⩾4000 g); 50 controls (normal weight, birth weight 2500-3999 g)]. Metabolites in umbilical cord blood were detected using an untargeted metabolomic approach based on gas chromatography/mass spectrometry. We performed logistic regression to evaluate the associations between metabolites and macrosomia. We also performed pathway analysis based on KEGG and MBRole. RESULTS: Compared with controls, the macrosomia cases had a greater male proportion, gestational age, paternal body mass index (BMI) and maternal pre-pregnancy BMI. Forty-two metabolites differed between the cases and controls. After multivariable adjustment, 2-methylfumarate [adjusted odds ratio (AOR)=1.232, 95% confidence interval (CI): 1.102-1.376], uracil (AOR=38.494, 95% CI: 5.635-262.951), elaidic acid (AOR=0.834, 95% CI: 0.761-0.915), ribose (AOR=0.089, 95% CI: 0.021-0.378), lactulose (AOR=0.815, 95% CI: 0.743-0.894) and 4-aminobutyric acid (AOR=0.835, 95% CI: 0.764-0.912) remained significantly associated with macrosomia. Pyrimidine metabolism and pentose and glucuronate interconversions were the two top-ranking pathways enriched with those metabolites (-log P-value=3.49 and 2.47, respectively). CONCLUSION: Levels of 2-methylfumarate, uracil, ribose, elaidic acid, lactulose and 4-aminobutyric acid were associated with the incidence of macrosomia. The alteration of pathways involving those factors might be linked with the incidence of macrosomia and relevant metabolic syndromes later in life, and further studies are needed to confirm it and verify the mechanisms.

[Benzoquinone induce autophagy in HL60 cells and the role of reactive oxygen species in induced autophagy].

OBJECTIVE: To investigate whether autophagy can be induced by 1, 4-benzoquinone (1, 4-BQ) in HL60 cells, as well as the role of reactive oxygen species (ROS) in induced autophagy. METHODS: In order to determine a suitable 1, 4-BQ treatment concentration for autophagy detection in HL60 cells, the cell vitality were examined by CCK8 assay. Logarithmic-growth-phased cells were divided into control group, 1, 4-BQ group (10µmol/L 1, 4-BQ, 24 h) , NAC group (antioxidant n-acetyl cysteine, 5mmol/L, 24 h) and the 1, 4-BQ+NAC group (5 mmol/L NAC were preincubated for 1h prior to the treatment with 10 µmol/L 1, 4-BQ for 24 h). The autophagic acidic vesicle were inspected by acridine orange staining, LC3 were detected by immunofluorescence staining, and expressions of LC3 and Beclin1 were quantitatively detected by Western blot. RESULTS: The results from cell viability test indicated that 1, 4-BQ exhibited a dose-dependent toxicity to HL60 cells. Compared with control group.the cell viability in 20.0、40.0µmol/L concentration were decreased obviously, and the differences had statistical significance (P<0.05). Compare with contrd group acidic vesicle, LC3II, LC3II/LC3I and Beclin1 protein expressions were increased in 1, 4-BQ group, after both respectively 12.4% and 27%, the differences had statistital significance. While 1, 4-BQ+NAC group was observed that acidic vesicle, LC3 and Beclin1 protein level were markedly lower than 1, 4-BQ group, after both decreased 12.6% and 22.6% respectively, both the difference were statistically significant (P<0.05). CONCLUSION: 1, 4-BQ can induce autophagy in HL60 cells, the induction of autophagy is at least partly resulted from ROS. Antioxidant can effectively suppress the occurrence of induced autophagy.

[A novel catalytic spectrophotometric determination for horseradish peroxidase].

A new catalytic-kinetics spectrophotometric method for the determination of horseradish peroxidase was developed. It was based on the catalytic effect of horseradish peroxidase (HRP) on the coloration reaction, in which the reduced rhodamine B was oxidized by H2O2 in pH 6.80 phosphate buffer. It was found that reduced rhodamine B stocking solution could be steady in 0.25% beta-CD solution. The kinetic behavior of the reaction and the effects of some experimental conditions were investigated and discussed in detail. The method has been applied to determine HRP with a satisfactory result. The calibration curve is linear over the range 15-250 pg.10 mL-1 of HRP (r = 0.9989). The limit of detection was 12 pg.10 mL-1 and the relative standard derivative RSD is 4.25% by determining for 20 pg.10 mL-1 HRP (n = 10).