Objective To evaluate thyroid‐stimulating immunoglobulin (TSI) and thyrotropin receptor antibodies (TRAb) diagnostic performance for Graves' disease (GD) and determine clinical cut‐off value for diagnosing GD. Methods Of 1369 retrospectively enrolled subjects, 1364 had a definitive diagnosis of untreated GD (GD‐UT, n = 87); treated GD (GD‐T, n = 206); autoimmune thyroid disease (AIT, n = 241); thyroid nodules (TN, n = 677); subacute thyroiditis (ST, n = 28); healthy subjects (HS, n = 125); other diseases with serological hyperthyroidism (n = 5) and were grouped into the following: UT‐GD and control groups (AIT, TN, ST, and HS); and UT‐GD and non‐GD hyperthyroidism groups. Diagnostic performance of TSI and TRAb was evaluated using area under the curve (AUC) of receiver‐operating characteristic (ROC) curve, and optimal clinical cut‐off value was determined using maximization of Youden index. Results TRAb AUC and clinical cut‐off value for diagnosing GD were 0.981 and 1.245 IU/L (sensitivity, 96.6%; specificity, 97.1%; positive predictive value [PPV], 71.8%; negative predictive value [NPV], 99.9%; positive likelihood ratio [PLR], 33.31; negative likelihood ratio [NLR, 0.035), respectively, for the GD‐UT and control groups. Those for TSI were 0.992 and 0.467 IU/L (sensitivity 98.8%; specificity, 96.4%; PPV, 68.8%; NPV, 99.9%; PLR, 27.472; NLR, 0.011). Those for TRAb in GD‐UT and non‐GD hyperthyroidism groups were 0.923 and 1.78 IU/L (sensitivity, 92.0%; specificity, 89.1%; PPV, 93%; NPV, 87.5%; PLR, 8.44; NLR, 0.089), respectively. For TSI, these were 0.92 and 0.545 IU/L (sensitivity, 97.7%; specificity, 83.6%; PPV, 90.4%; NPV, 95.8%; PLR27.472, NLR, 0.011), respectively. Conclusion TSI diagnostic performance for GD was excellent and had better sensitivity than TRAb.

Purpose Human follicular fluid (FF) is a complex biological fluid that contributes to the microenvironment of oocyte development. The aim of this study was to evaluate the role of steroid levels and mitochonrial function in embryo development during in vitro fertilization cycles. Methods This was a cohort study of 138 women receiving IVF/ICSI, including 136 FF samples from 109 infertile women. FF steroid levels were tested by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and immunoassays. The mRNA expression levels of mitochondrial electron transport chain (ETC) complex genes from FF exosomes were detected by qPCR. Results Analysis of these individual FF concentrations revealed that LH and FSH concentrations were higher in follicles in which the oocyte developed into a top quality (TQ) blastocyst (LH: 9.44 ± 2.32mIU/ml, FSH: 9.32 ± 1.01mIU/ml) than those in which there was a failure of fertilization (LH: 5.30 ± 0.84mIU/ml, FSH: 6.91 ± 0.62mIU/ml). In contrast, follicular cortisone concentrations were the lower for oocytes that resulted in a TQ blastocyst (12.20 ± 0.82mIU/ml). The receiver operating characteristic analysis showed that FF LH and FSH levels predicted TQ blastocyst with excellent AUC value of 0.711 and 0.747. Mitochondrial ETC complex I and III mRNA levels were increased in the FF exosomes of TQ blastocyst. Correlation analysis showed that mRNA levels of ETC complex I was positively correlated with LH and FSH levels in FF. Conclusion The levels of FF steroids from single follicle can predetermine subsequent embryo development to some extent. Furthermore, impaired exosome mitochondrial dysfunction is a potiential event that causes hormone change in embryo development.