Performance of LC/MS/MS in Analyzing Multiple Amino Acids and Acylcarnitines in Dried Blood Spot of Very Low Birth Weight, Low Birth Weight, and Normal Weight Neonates.

BACKGROUND: Amino acid (AA) and acylcarnitine (AC) are important biomarkers of protein and fatty acid metabolism. Examining their levels in newborns may reveal multiple inherited metabolic diseases. However, they have rarely been assessed in very low birth weight (VLBW) neonates, low birth weight (LBW) neonates and rarely been compared with normal weight (NW) neonates. The aim of the study was to identify the AA and AC profiles in dried blood spot (DBS) specimens of LBW and VLBW neonates, then compare with NW neonates, and make a contribution to the determination of cutoff values of VLBW and LBW neonates. METHODS: Liquid Chromatography tandem mass spectrometry (LC/MS/MS) is an excellent tool for quantitatively detecting AA and AC profiles. This article verified the precision, accuracy, and linearity of the LC/MS/MS method in AA and AC detection, then analyzed AA and AC profiles in DBS of VLBW, LBW and NW neonates, and compared the difference of AA and AC in the three groups. RESULTS: The results showed that the LC/MS/MS method had wide linear range, satisfied precision and reproducibility in detecting AA and AC in DBS specimens; most AA and AC concentrations significantly correlated with birth weight in DBS samples (p < 0.05). CONCLUSIONS: The results suggested that VLBW and LBW neonates have different metabolic or nutritional status with NW neonates and different AA and AC cutoffs should be defined for them to reduce the risk of false-positive cases.

[Effect of bushen tiaojing recipe on the quality of the oocytes and reproductive hormones in the follicular fluid in IVF-ET patients].

OBJECTIVE: To explore the effect of Bushen Tiaojing Recipe (BTR) on the quality of oocytes, reproductive hormones, and the expression of bone morphogenetic protein-15 (BMP15) of in vitro fertilization-embryo transfer (IVF-ET) patients. METHODS: Sixty infertility patients who prepared for IVF-ET were assigned to two groups according to the treatment order, the treatment group [20 cases, treated with BTR + controlled ovarian hyperstimulation (COH)] and the control group (treated with COH alone, 40 cases). Age, the time limit for infertility, basal follicle-stimulating hormone (bFSH) concentration, usage days and the dosage of gonadotropins (Gn), serum levels of estradiol (E2), luteotropic hormone (LH), and progesterone (P) on the HCG injection day, the number of retrieved occytes, the fertilization rate, the number of embryos, the high quality embryo rate, and the clinical pregnancy rate were compared. Concentrations of follicle-stimulating hormone (FSH), LH, E2, testosterone (T), and P in the follicular fluid were detected via chemiluminescence microparticle immunoassay. The mRNA and protein expression of BMP-15 in mature granulosa cells was detected by real-time fluorescent PCR and Western blot. RESULTS: Thirty-two patients were pregnant and the total pregnancy rate was 53.3%. Of them, 19 were pregnant and the total pregnancy rate was 47.5% in the control group, while 20 were pregnant and the total pregnancy rate was 65.0% in the treatment group. But there was no statistical difference between the two groups (P > 0.05). Compared with the control group, the Gn dosage was lower and the high quality embryo rate was higher in the treatment group, showing statistical difference (P < 0.05). There was no statistical difference in serum concentrations of E2, LH, or P on the HCG injection day, the number of retrieved oocytes, or the fertilization rate (P > 0.05). Compared with the control group, FSH concentrations in the follicular fluid were significantly lower and LH concentrations were significantly higher in the treatment group (P < 0.05). The LH concentrations in the follicular fluid were significantly higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).There was no statistical difference in E2, T, or P concentrations (P > 0.05). The mRNA and protein expression of BMP-15 in granulosa cells was higher in the treatment group than in the control group (P < 0.05). It was also higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05). CONCLUSION: During the IVF-ET process, BTR could elevate the quality of oocytes, and increase the sensitivity of ovarian follicles to exogenous Gn, which was correlated with the mRNA and protein expression of BMP-15 in granulosa cells, and changing concentrations of FSH and LH.