Predicting risk of blastocyst aneuploidy among women with previous aneuploid pregnancy loss: a multicenter-data-based multivariable model.

STUDY QUESTION: Can blastocyst aneuploidy be predicted for patients with previous aneuploid pregnancy loss (PAPL) and receiving preimplantation genetic testing for aneuploidy (PGT-A)? SUMMARY ANSWER: Multivariable logistic regression models were established to predict high risk of blastocyst aneuploidy using four identified factors, presenting good predictive performance. WHAT IS KNOWN ALREADY: Aneuploidy is the most common embryonic chromosomal abnormality leading to pregnancy loss. Several studies have demonstrated a higher embryo aneuploidy rate in patients with PAPL, which has suggested that PGT-A should have benefits in PAPL patients intending to improve their pregnancy outcomes. However, recent studies have failed to demonstrate the efficacy of PGT-A for PAPL patients. One possible way to improve the efficacy is to predict the risk of blastocyst aneuploidy risk in order to identify the specific PAPL population who may benefit from PGT-A. STUDY DESIGN, SIZE, DURATION: We conducted a multicenter retrospective cohort study based on data analysis of 1119 patients receiving PGT-A in three reproductive medical centers of university affiliated teaching hospitals during January 2014 to June 2020. A cohort of 550 patients who had one to three PAPL(s) were included in the PAPL group. In addition, 569 patients with monogenic diseases without pregnancy loss were taken as the non-PAPL group. PARTICIPANTS/MATERIALS, SETTING, METHODS: PGT-A was conducted using single nucleotide polymorphism microarrays and next-generation sequencing. Aneuploidy rates in Day 5 blastocysts of each patient were calculated and high-risk aneuploidy was defined as a rate of ≥50%. Candidate risk factors for high-risk aneuploidy were selected using the Akaike information criterion and were subsequently included in multivariable logistic regression models. Overall predictive accuracy was assessed using the confusion matrix, discrimination by area under the receiver operating characteristic curve (AUC), and calibration by plotting the predicted probabilities versus the observed probabilities. Statistical significance was set at P < 0.05. MAIN RESULTS AND THE ROLE OF CHANCE: Blastocyst aneuploidy rates were 30 ± 25% and 21 ± 19% for PAPL and non-PAPL groups, respectively. Maternal age (odds ratio (OR) = 1.31, 95% CI 1.24-1.39, P < 0.001), number of PAPLs (OR = 1.40, 95% CI 1.05-1.86, P = 0.02), estradiol level on the ovulation trigger day (OR = 0.47, 95% CI 0.30-0.73, P < 0.001), and blastocyst formation rate (OR = 0.13, 95% CI 0.03-0.50, P = 0.003) were associated with high-risk of blastocyst aneuploidy. The predictive model based on the above four variables yielded AUCs of 0.80 using the training dataset and 0.83 using the test dataset, with average and maximal discrepancies of 2.89% and 12.76% for the training dataset, and 0.98% and 5.49% for the test dataset, respectively. LIMITATIONS, REASONS FOR CAUTION: Our conclusions might not be compatible with those having fewer than four biopsied blastocysts and diminished ovarian reserves, since all of the included patients had four or more biopsied blastocysts and had exhibited good ovarian reserves. WIDER IMPLICATIONS OF THE FINDINGS: The developed predictive model is critical for counseling PAPL patients before PGT-A by considering maternal age, number of PAPLs, estradiol levels on the ovulation trigger day, and the blastocyst formation rate. This prediction model achieves good risk stratification and so may be useful for identifying PAPL patients who may have higher risk of blastocyst aneuploidy and can therefore acquire better pregnancy outcomes by PGT-A. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the National Natural Science Foundation of China under Grant (81871159). No competing interest existed in the study. TRIAL REGISTRATION NUMBER: N/A.

[Effect of lutein on relieving oxidative stress in mice induced by D-galatose].

OBJECTIVE: To study the effect of lutein on relieving oxidative stress in the liver of mice induced by D-galactose(D-gal). METHODS: Forty eight Kunming strain mice were randomized into 4 groups: model group, low lutein group (LL 10 mg/(kg x d)), high lutein group (HL 40 mg/(kg x d)) and normal control group. The content of reactive oxygen species (ROS), nitric oxide (NO) and activity of total nitric oxide synthase (TNOS), inducible nitric oxide synthase (iNOS) and mitochondrial Na(+)-K(+)-ATPase, Ca(2+)-ATPase in liver tissue were detected 6 weeks later in the experiment. The expression of toll-like receptor-4 (TLR4) mRNA and hemeoxygenase-1 (HO-1) mRNA in hepatic tissue were detected by reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: ROS content in HL and LL group was significantly lower (P < 0.01) than that in the model group. The activity of Na(+)- K(+)-ATPase in HL and LL group and the activity of Ca(2+)-ATPase in HL group were significantly higher than that in the model group (P < 0.05). The activities of TNOS and iNOS and the content of NO in HL group were significantly lower than the model group (P < 0.05). The expression of HO-1 mRNA in HL group was significantly higher than that in the model group, but the expression of TLR4 mRNA in HL group was significantly lower than that in the model group (P < 0.05). CONCLUSION: The mechanism of lutein on the protection of mice from oxidative stress induced by D-gal might be related to increasing the expression of HO-1 mRNA and reducing the expression of TLR4 mRNA.

[Effects of lycopene on cerebral ischemia-reperfusion injury in rats].

OBJECTIVE: To study the protective effects of lycopene (LP) on cerebral ischemia-reperfusion injury and oxidative stress in SD rats and the mechanism of them. METHODS: The rats were divided into five groups: normal control group, model control group, sham group and two LP groups (fed with 5 mg/kg bw or 20 mg/kg bw of lycopene daily for 15 days). The model for cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion (MCAO). The score of neurological behavior was evaluated at the 3rd and 24th hours after reperfusion. The rats were put to death 24 h after reperfusion. The size of cerebral infarction was measured. The activities of iNOS, SOD, CAT and the contents of NO and MDA in brain and serum uric acid were measured. The expressions of Bcl-2 mRNA and HIF-1alphamRNA in cortex were examined by using reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: In comparison with the model group, the neurological deficits were milder, the volumes of cerebral infarction were smaller, the activities of SOD, CAT in brain tissue were higher, the activities of iNOS as well as the contents of NO, MDA in brain tissue and serum uric acid were lower in Lycopene groups. Compared with the model group and control group, the expression of HIF-1 alpha mRNA of cortex in the high dose lycopene (20 mg/kg bw) group was up-regulated; while the expression of Bcl-2 mRNA of cortex was up-regulated only in the low dose lycopene (5 mg/kg bw) group. CONCLUSION: There were some protective effects of oral administration of lycopene against cerebral ischemia-reperfusion injuries induced by focal cerebral ischemia and oxidative stress. The possible mechanism may be related with increasing activities of antioxidant enzymes, inhibiting lipid peroxidation, decreasing activities of iNOS,and up-regulating the expression of HIF-1alpha mRNA as well as Bcl-2 mRNA.

[The effects of lycopene on reactive oxygen species and anoxic damage in ischemia reperfusion injury in rats].

OBJECTIVE: To study the protective effects of lycopene (LP) on cerebral ischemia-reperfusion injury induced by focal cerebral ischemia and oxidative stress in rats. METHODS: 48 male Sprague-Dawley (SD) rats were randomly assigned into five groups: A (20 mg/kg LP), B (5 mg/kg LP), C (salad oil), D (salad oil) and E (basic feed control). A, B and C groups were given LP or salad oil orally for 15 d, then cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion (MCAO) and D group was used as fake surgery control. The contents of reactive oxygen species (ROS), nitric oxide (NO), lactic acid (LD) and the activities of nitric oxide synthetase (NOS) in cortex were measured at 24 h after reperfusion. The levels of HIF-1alpha mRNA and Bcl-2 mRNA in hippocampi were determined by using reverse transcription polymerase chain reaction (RT- PCR) technique. RESULTS: ROS levels of A, B, C, D and E groups were (114.23 +/- 18.91), (135.89 +/- 14.17), (171.37 +/- 25.76), (94.24 +/- 2.23) and (92.06 +/- 5.59) fluorescence intensity value/g protein, respectively (F = 9.038, P < 0.01); levels of NO were (6.60 +/- 0.77), (7.13 +/- 0.47), (8.38 +/- 0.80), (5.52 +/- 0.16) and (5.23 +/- 0.51) micromol/g protein respectively (F = 10.197, P < 0.01); levels of NOS were (0.817 +/- 0.016), (0.875 +/- 0.095), (1.030 +/- 0.101), (0.557 +/- 0.094) and (0.595 +/- 0.066) U/mg protein respectively (F = 14.555, P < 0.01); levels of LD were (0.381 +/- 0.069), (0.446 +/- 0.012), (0.576 +/- 0.059), (0.359 +/- 0.021) and (0.310 +/- 0.036) mmol/g protein respectively (F = 10.043, P < 0.01); HIF-1alpha mRNA expression levels in hippocampi were 0.865 +/- 0.274, 0.635 +/- 0.069, 0.491 +/- 0.067, 0.375 +/- 0.052 and 0.361 +/- 0.087, respectively (F = 40.520, P < 0.01); and Bcl-2 mRNA expression levels in hippocampi were 0.263 +/- 0.033, 0.330 +/- 0.028, 0.198 +/- 0.034, 0.304 +/- 0.039 and 0.236 +/- 0.025, respectively (F = 11.003, P < 0.01). CONCLUSION: The protective effects of LP may be related with its abilities of decreasing ROS and LD cumulation, alleviating inflammation and up-regulating the expression of protective genes.