Objective: To evaluate estrogen and progesterone levels' impact and combined effects on clinical pregnancy rate and live birth rate in gonadotropin-releasing hormone antagonist regimen with fresh embryo transfer on human chorionic gonadotropin (hCG) trigger day. Methods: The data of patients who underwent in vitro fertilization or intracytoplasmic sperm injection at The First Affiliated Hospital of Nanjing Medical University from January 2011 to January 2021 were collected. According to the number of retrieved oocytes, they were divided into the normal response group (5-15 oocytes) and the low response group (<5 oocytes). The ovulation induction outcomes and pregnancy outcomes of the two groups were compared. Statistical analysis was conducted using t test or chi-square test, and confounding factors were adjusted through multivariate regression models, and the hormone cut-off values were determined. Results: Among 3 595 patients, compared with the normal response group (n=2 614), the age of women in the low response group (n=981) had a negative impact on pregnancy outcomes (P<0.001), while the number of antral follicles, anti-Müllerian hormone, endometrial thickness and embryo quality were positively correlated with pregnancy outcomes (all P<0.05). Multivariate regression analysis indicated that in the normal response group, a higher level of estradiol on hCG trigger day were significantly associated with decreases in clinical pregnancy rate (OR=0.965, 95%CI: 0.946-0.984; P<0.001) and live birth rate (OR=0.973, 95%CI: 0.954-0.993; P=0.008). In the low response group, higher progesterone level was associated with poorer pregnancy outcomes (clinical pregnancy rate: OR=0.875, 95%CI: 0.794-0.961, P=0.006; live birth rate: OR=0.876, 95%CI: 0.793-0.966, P=0.009). The hormone cut-off values: the estradiol cut-off values for the normal and low response groups were 14 000 pmol/L and 6 000 pmol/L respectively, and the progesterone cut-off values for the normal and low response groups were 5.8 nmol/L and 4.0 nmol/L. When hormone levels exceed the cut-off values, pregnancy outcomes significantly decline, and the negative impact on pregnancy outcomes was most significant when both hormones increase (clinical pregnancy rate: normal response group P=0.010，low response group P=0.020; live birth rate: normal response group P=0.028，low response group P=0.014). Conclusions: In patients with normal response, compared with progesterone on hCG trigger day, the increase in estradiol significantly reduces the clinical pregnancy rate and live birth rate of fresh embryo transfer, which should be paid attention to. In patients with poor response, both higher progesterone and estradiol levels on hCG trigger day could significantly reduce the clinical pregnancy rate and live birth rate of fresh embryo transfer, and the combined effect is more obvious. It is recommended that for patients whose progesterone and estradiol on the hCG day exceed the corresponding cut-off values, all embryos should be frozen and embryo transfer should be postponed.
Objective: To evaluate the clinical application value of practical clinical classification system for cesarean scar pregnancy (CSP). Methods: A total of 1 076 patients with CSP admitted to Peking University Third Hospital from January 2014 to December 2023 were retrospectively included in the study. According to the practical clinical classification system for CSP, the patients were divided into 5 groups (type Ⅰ, Ⅱa, Ⅱb, Ⅲa and Ⅲb). The clinical characteristics, diagnosis and treatment variance of different types of CSP patients were analyzed, and the treatment strategy recommended by this study was compared with that recommended by the practical clinical classification system. Results: The prevalence of type Ⅰ, Ⅱa, Ⅱb, Ⅲa and Ⅲb were 37.1% (399/1 076), 42.0% (452/1 076), 8.5% (91/1 076), 10.5% (113/1 076) and 2.0% (21/1 076), respectively. The median gestational age at diagnosis was 47 days, the median length of hospital stay was 2 days, the median hospitalization cost was 5 816 yuan, the median operation time was 36 minutes, the median intraoperative blood loss was 23 ml, and 5.6% (60/1 076) patients had intraoperative blood loss≥300 ml; 2.6% (28/1 076) patients received blood transfusion, 94.9% (1 021/1 076) patients underwent minimally invasive surgery. There were significant differences in the above indicators between the 5 groups (all P<0.05). The treatments of 86.0% (925/1 076) patients were consistent with the recommendations of the practical clinical classification system. There were 7.7% (83/1 076) patients had more minimally invasive treatment compared with the recommendations, all of which were type Ⅲ patients. There were 6.3% (68/1 076) patients had more invasive treatment compared with the recommendations, mainly were type Ⅱa, Ⅱb and Ⅲa patients. The first-line treatment was successful in 96.4% (1 037/1 076) patients. Conclusions: Practical clinical classification system for CSP has good discrimination and clinical applicability, and provides the principle of minimally invasive surgery. For the management of type Ⅲ patients, individualized evaluation should be carried out on the basis of flexible application of the practical clinical classification system, combined with the diagnosis and treatment conditions of medical institutions and the patient's condition.
Objective:To investigate the inhibitory effects of phloroglucinol on eutopic stromal cells and ectopic stromal cells from endometriosis and its molecular mechanism.Methods:The effect of phloroglucinol on the survival rate of endometriosis-derived eutopic stromal cells hEM15A and ectopic stromal cells ihESCs was detected by cell counting kit 8 method. The effect of phloroglucinol on cell migration were evaluated by scratch test and Transwell migration test. The level of reactive oxygen species (ROS) in cells was detected by the fluorescence probe method. The expression of nuclear factor erythroid 2-related factor 2 (NRF2), kelch like ECH associated protein 1 (KEAP1), glutathione peroxidase 4 (GPX4) proteins in KEAP1/NRF2/GPX4 pathway was detected by western blot.Results:(1) The results of cell counting kit 8 method showed that phloroglucinol significantly reduced the cell viability of hEM15A and ihESCs [(47.89±1.93)% and (81.55±6.16)% after treatment with 5.550 mmol/L for 48 hours, (18.44±6.64)% and (66.09±1.63)% after treatment with 5.550 mmol/L for 72 hours, (23.20±4.95)% and (51.05±7.16)% after treatment with 11.100 mmol/L for 48 hours, (3.62±0.23)% and (40.14±2.63)% after treatment with 11.100 mmol/L for 72 hours; all P<0.05]. (2) Scratch test results showed that the cell migration rates of hEM15A cells and ihESCs cells at 24 hours time points without phloroglucinol (0 mmol/L) were (41.02±2.85)% and (41.60±1.77)%, respectively. Compared with the 0 mmol/L group of the respective cells, the migration rates of hEM15A and ihESCs cells treated with different concentrations (2.775, 5.550, 8.325, 11.100, 13.875 mmol/L) of phloroglucinol at 24 hours were significantly decreased [hEM15A cells were (21.28±2.96)%, (9.97±3.42)%, (3.39±0.79)%, (3.26±0.47)% and (2.38±0.11)%, ihESCs cells were (20.31±2.17)%, (17.17±1.44)%, (3.68±0.94)%, (2.96±1.19)% and (2.57±0.43)%; all P<0.05]. (3) The results of Transwell migration test showed that the numbers of migrated hEM15A and ihESCs cells were 196.00±14.00 and 73.67±5.51, respectively, at 24 hours without phloroglucinol (0 mmol/L) treatment. Compared with the respective 0 mmol/L group of the respective cells, there were no significant changes in the numbers of migration of the two cells in the 2.775 mmol/L group at 24 hours (190.30±42.59 and 68.33±16.56, respectively; all P>0.05). After treatment with the remaining higher concentration of phloroglucinol, the migration ability of hEM15A and ihESCs cells were significantly reduced (hEM15A cells were 131.70±29.96, 117.70±4.51, 83.33±12.42 and 42.33±13.87, ihESCs cells were 42.33±8.51, 41.00±9.54, 18.33±7.37 and 15.67±2.52; all P<0.05). (4) The results of ROS level detected by the fluorescence probe method showed that there were no significant changes in ROS levels in hEM15A cells and ihESCs cells after treatment with 2.775 mmol/L phloroglucinol for 24 hours (1.21±0.12 and 1.16±0.11, respectively; all P>0.05). After treatment for 48 and 72 hours, the levels of ROS in the two kinds of cells were significantly increased (2.75±1.15 and 2.76±0.74 in hEM15A cells, 2.82±0.72 and 2.90±0.82 in ihESCs cells; all P<0.05). (5) Western blot results showed that the expressions of KEAP1 in hEM15A cells and ihESCs cells were significantly decreased after phloroglucinol treatment (0.34±0.16 and 0.33±0.08; all P<0.05), and the expressions of NRF2 and GPX4 were significantly up-regulated (hEM15A cells were 2.88±0.44 and 2.09±0.06, ihESCs cells were 3.43±0.49 and 1.73±0.14; all P<0.05). Conclusion:Phloroglucinol could affect the survival of eutopic stromal cells and ectopic stromal cells from endometriosis by inducing oxidative stress, and its mechanism may be related to the activation of KEAP1/NRF2/GPX4 pathway and the imbalance of redox homeostasis.
Objective:To evaluate the efficacy and safety of hexaminolevulinate photodynamic therapy (HAL-PDT) for the treatment of cervical intraepithelial neoplasia 2 (CIN2).Methods:Data of this study were derived from an international multicenter, randomized, double-blind, placebo-controlled phase Ⅲ clinical trial (the APRICITY study) conducted between October 2020 and July 2023, with medical records obtained from 61 medical centers across 7 countries. A subgroup analysis was performed in patients with histologically confirmed CIN2 from this trial. Participants were randomly assigned in a 2∶1 ratio to the HAL-PDT group [5% hexaminolevulinate ointment and a light-emitting diode (LED) light source device] or the placebo group (placebo ointment with a non-illuminating device). The primary endpoint was the response rate at 6 months after the initial treatment. Secondary endpoints included histological regression rate, high-risk human papillomavirus (HR-HPV) clearance rate. Safety endpoints included treatment-related adverse events (TRAEs).Results:A total of 182 patients with CIN2 were included (120 in the HAL-PDT group, 62 in the placebo group). Baseline characteristics were balanced between the two groups (all P>0.05). At 6 months, the response rate [49.2% (59/120) vs 22.6% (14/62), P=0.001] and histological regression rates [57.5% (69/120) vs 30.6% (19/62), P=0.001] in the HAL-PDT group were significantly higher than those in the placebo group. The colposcopic lesion clearance rate at 6 months was also higher in the HAL-PDT group than in the placebo group [60.6% (60/99) vs 32.7% (17/52), P=0.001]. The risk of disease progression was lower in the HAL-PDT group ( RR=0.55, 95% CI:0.29-1.06; P=0.068). Analyses stratified by age, HR-HPV infection status, region, and lesion size consistently demonstrated benefit in the HAL-PDT group regarding response and histological regression rates. At the 12-month follow-up, HR-HPV clearance rates in the HAL-PDT group were significantly improved, reaching 45.9% for any HR-HPV subtype, 58.8% for HPV 16, and 55.4% for HPV 16/18. There was no statistically significant difference in the incidence of TRAEs between the two groups [31.1% (37/119) vs 25.8% (16/62), P=0.496]. The most commonly reported adverse events in the HAL-PDT group were increased vaginal discharge, abdominal pain, and vulvovaginal pain, all of which were mild and self-limiting. Conclusion:HAL-PDT demonstrated favorable efficacy and safety in patients with CIN2, providing a non-surgical treatment alternative for patients seeking fertility preservation and the maintenance of cervical integrity.
