The expression of High-temperature requirement factor A4 (HtrA4) mRNA is significantly lower in the chorionic villi of patients with recurrent pregnancy loss (RPL) than in the control group. We conducted an investigation into the cellular functions of HtrA4 using the CRISPR/Cas9 system and shRNA-HtrA4 to create knockout BeWo cells and HtrA4 knockdown JEG3 cells. Our results indicated that the knockout BeWo cells exhibited reduced capacity for invasion and fusion, but increased levels of proliferation and migration, with a significantly shortened cell cycle compared to wild-type cells. Wild-type BeWo cells highly expressed cell invasion- and fusion-related factors, while knockout BeWo cells highly expressed migration-, proliferation-, and cell cycle-related factors. The shRNA-HtrA4 JEG3 cells showed a decreased capacity for invasion, but an increased capacity for migration, accompanied by a decrease in the expression of cell invasion-related factors and an increase in migration-related factors. Moreover, our ELISA results revealed that the serum HtrA4 level was lower in patients with RPL than in the controls. These findings suggest that HtrA4 depletion may be associated with placental dysfunction.

Objectives: This study aimed to compare the impact of robotic (RO) versus laparoscopic single-site ovarian cystectomy (LO) on ovarian reserve, as measured by changes in anti-Müllerian hormone (AMH) levels. Methods: A total of 43 women undergoing LO and 40 women undergoing RO for mature cystic teratoma were included. The baseline characteristics and surgical outcomes were scrutinized. AMH levels were evaluated preoperatively and at 3 months after surgery. Results: BMI (body mass index) was higher in the LO group (23.1 ± 3.6 cm vs. 21.7 ± 2.1 kg/m², p = 0.03) than in the RO group. Otherwise, there were no statistically significant differences in patient characteristics. The LO group showed a shorter operative time (70.0 ± 24.0 vs. 86.5 ± 26.7 min, p = 0.002) than the RO group., However, multiple linear regression analysis revealed that BMI was independently associated with increased operative time (p = 0.001), while the surgical method was not a significant predictor (p = 0.725). There were no significant differences in the rate of decline in AMH level between the LO and RO groups (13.3 ± 21.2 vs. 4.64 ± 34.1%, p = 0.167). There were intergroup differences in the hemostasis method: in the LO group, bipolar coagulation was performed for 27 (62.8%) patients, and sutures were performed for 16 (37.2%) patients; in the RO group, bipolar coagulation was performed for 5 (12.5%) patients, and sutures were performed for 33 (82.5%) patients, while in the case of 2 (5.0%) patients, hemostasis was not performed (p < 0.001). However, note that in our logistic regression analysis, the hemostasis method was excluded as an independent factor affecting AMH. Conclusions: There appear to be no significant advantages of RO over LO in terms of ovarian reserve preservation.