Objective:To investigate the protective effects of resveratrol (RSV) on human sperm cryopreservation and explore its underlying protective mechanisms.Methods:A total of 165 normal fresh semen samples were collected from the Reproductive Medicine Center, Department of Obstetrics and Gynecology and Human Sperm Bank of the First Affiliated Hospital of Anhui Medical University between December 2022 and December 2024. Among them, 65 samples were used to obtain semen parameters before and after conventional freezing. Each sample of the other 104 samples was mixed at a 2∶1 volume ratio with cryoprotectant containing 0, 10 -?, 10 -?, or 10 -? mol/L RSV, followed by cryopreservation in liquid nitrogen for 24 h. Post-thaw assessments included routine sperm parameters, sperm DNA fragmentation index (DFI) evaluated by sperm chromatin dispersion assay, reactive oxygen species (ROS) levels measured via flow cytometry, RSV and nuclear factor erythroid 2-related factor 2 (NRF2) interactions examined by molecular docking and cellular thermal shift assay (CETSA), NRF2 protein contents analyzed by immunofluorescence and Western blotting, mRNA levels of NRF2 and downstream antioxidant proteins Heme Oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1) quantified by qRT-PCR and effects of NRF2 inhibitor ML385 on sperm parameters. Results:Compared with fresh samples, conventional cryopreservation significantly reduced sperm motility (all P<0.001). The addition of 10 -? mol/L RSV significantly improved the percentage of forward motile sperm after freezing (26.98%±8.98% vs. 19.61%±8.03%, P<0.001) while reducing DFI (9.84%±3.81% vs. 15.06%±4.22%, P<0.001) and ROS levels ( P<0.001) compared with the post-freezing group without the addition of RSV. Both molecular docking analysis and CETSA confirmed that RSV interacted with NRF2. Notably, sperm cryopreserved with 10 -? mol/L RSV exhibited significantly higher contents of NRF2 and its downstream effectors HO-1 and NQO1 compared with the post-freezing group without the addition of RSV (all P<0.001). This protective effect was markedly attenuated by co-treatment with the NRF2 inhibitor ML385, as evidenced by a significant decline in sperm motility ( P<0.001). Conclusion:RSV exerts cryoprotective effects likely through NRF2-mediated antioxidant pathways, reducing oxidative stress and enhancing post-thaw sperm quality.

Objective:To investigate the protective effects of resveratrol (RSV) on human sperm cryopreservation and explore its underlying protective mechanisms.Methods:A total of 165 normal fresh semen samples were collected from the Reproductive Medicine Center, Department of Obstetrics and Gynecology and Human Sperm Bank of the First Affiliated Hospital of Anhui Medical University between December 2022 and December 2024. Among them, 65 samples were used to obtain semen parameters before and after conventional freezing. Each sample of the other 104 samples was mixed at a 2∶1 volume ratio with cryoprotectant containing 0, 10 -?, 10 -?, or 10 -? mol/L RSV, followed by cryopreservation in liquid nitrogen for 24 h. Post-thaw assessments included routine sperm parameters, sperm DNA fragmentation index (DFI) evaluated by sperm chromatin dispersion assay, reactive oxygen species (ROS) levels measured via flow cytometry, RSV and nuclear factor erythroid 2-related factor 2 (NRF2) interactions examined by molecular docking and cellular thermal shift assay (CETSA), NRF2 protein contents analyzed by immunofluorescence and Western blotting, mRNA levels of NRF2 and downstream antioxidant proteins Heme Oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1) quantified by qRT-PCR and effects of NRF2 inhibitor ML385 on sperm parameters. Results:Compared with fresh samples, conventional cryopreservation significantly reduced sperm motility (all P<0.001). The addition of 10 -? mol/L RSV significantly improved the percentage of forward motile sperm after freezing (26.98%±8.98% vs. 19.61%±8.03%, P<0.001) while reducing DFI (9.84%±3.81% vs. 15.06%±4.22%, P<0.001) and ROS levels ( P<0.001) compared with the post-freezing group without the addition of RSV. Both molecular docking analysis and CETSA confirmed that RSV interacted with NRF2. Notably, sperm cryopreserved with 10 -? mol/L RSV exhibited significantly higher contents of NRF2 and its downstream effectors HO-1 and NQO1 compared with the post-freezing group without the addition of RSV (all P<0.001). This protective effect was markedly attenuated by co-treatment with the NRF2 inhibitor ML385, as evidenced by a significant decline in sperm motility ( P<0.001). Conclusion:RSV exerts cryoprotective effects likely through NRF2-mediated antioxidant pathways, reducing oxidative stress and enhancing post-thaw sperm quality.

Objective:To investigate the clinical outcomes of patients with multiple morphological abnormalities of the flagella (MMAF) caused by CFAP43 or CFAP44 gene mutations following intracytoplasmic sperm injection (ICSI). Methods:Clinical data and genetic information were retrospectively analyzed for 121 MMAF patients who attended Reproductive Medicine Center, Department of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical University during September 2014 to July 2020. Totally 9 MMAF patients were identified to carry CFAP43 or CFAP44 mutations, 5 of them (P3, P5, P7, P8, and P9) received ICSI treatments, the ICSI outcomes were further analyzed. Results:Sanger sequencing validated 9 MMAF patients harboring CFAP43 or CFAP44 biallelic mutations, our study firstly identified a novel homozygous mutation of CFAP43(c.4132delC: p.Arg1378Glufs*10), novel compound heterozygous mutations of CFAP43 (c.3938G>A: p.Arg1313Gln;c.4342G>A:p.Glu1448Lys) and novel compound heterozygous mutations of CFAP44 (c.1718C>A:p.Pro573His; c.4075G>A: p.Glu1359Lys). The 5 MMAF patients underwent 5 ICSI cycles, 4 healthy offspring were obtained. The rate of fertilization of CFAP43- or CFAP44-mutated MMAF patients following ICSI was 76.47% (39/51), 3 patients' wife got clinical pregnancy, 3 patients got live birth delivery, respectively. No significant differences were found in ICSI outcomes among CFAP43-mutated or CFAP44-mutated MMAF patients, DNAH1-mutated MMAF patients, and severe oligoasthenozoospermia group (all P>0.05) .Conclusion:CFAP43 or CFAP44 mutations are responsible for the malformation of sperm flagella and decrease of sperm motility, and validated as the important genetic causes of MMAF. CFAP43- or CFAP44-mutated MMAF patients could have a favorable treatment outcome following ICSI.

Objective:To investigate the clinical outcomes of patients with multiple morphological abnormalities of the flagella (MMAF) caused by CFAP43 or CFAP44 gene mutations following intracytoplasmic sperm injection (ICSI). Methods:Clinical data and genetic information were retrospectively analyzed for 121 MMAF patients who attended Reproductive Medicine Center, Department of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical University during September 2014 to July 2020. Totally 9 MMAF patients were identified to carry CFAP43 or CFAP44 mutations, 5 of them (P3, P5, P7, P8, and P9) received ICSI treatments, the ICSI outcomes were further analyzed. Results:Sanger sequencing validated 9 MMAF patients harboring CFAP43 or CFAP44 biallelic mutations, our study firstly identified a novel homozygous mutation of CFAP43(c.4132delC: p.Arg1378Glufs*10), novel compound heterozygous mutations of CFAP43 (c.3938G>A: p.Arg1313Gln;c.4342G>A:p.Glu1448Lys) and novel compound heterozygous mutations of CFAP44 (c.1718C>A:p.Pro573His; c.4075G>A: p.Glu1359Lys). The 5 MMAF patients underwent 5 ICSI cycles, 4 healthy offspring were obtained. The rate of fertilization of CFAP43- or CFAP44-mutated MMAF patients following ICSI was 76.47% (39/51), 3 patients' wife got clinical pregnancy, 3 patients got live birth delivery, respectively. No significant differences were found in ICSI outcomes among CFAP43-mutated or CFAP44-mutated MMAF patients, DNAH1-mutated MMAF patients, and severe oligoasthenozoospermia group (all P>0.05) .Conclusion:CFAP43 or CFAP44 mutations are responsible for the malformation of sperm flagella and decrease of sperm motility, and validated as the important genetic causes of MMAF. CFAP43- or CFAP44-mutated MMAF patients could have a favorable treatment outcome following ICSI.

Objective: To investigate the short-term outcome of patients with acute myocardial infarction complicating cardiogenic shock due to left main disease. Methods: A total of 24 patients with acute myocardial infarction complicating cardiogenic shock due to left main artery disease hospitalized in Fuwai hospital from June 2012 to May 2018 were included. The clinical data were analyzed,and the patients were divided into survivor group (11 cases) and death group (13 cases) according to survival status at 28 days post the diagnosis of shock. The patients were further divided into thrombolysis in myocardial infarction(TIMI) flow grade 0-2 group (11 cases) and TIMI flow grade 3 group (13 cases) according to TIMI flow grade after the procedure. The patients were then divided into non-three-vessel lesions group (14 cases) and three-vessel lesions group (10 cases) according to coronary angiography results. Results: Compared with survivor group, patients in death group presented with lower worst systolic blood pressure within 24 hours after admission (50(48, 70) mmHg (1 mmHg=0.133 kPa) vs. 73(70, 80) mmHg, P<0.01), lower worst diastolic blood pressure with in 24 hours after admission ((33.5±12.4) mmHg vs. (48.9±9.4) mmHg, P<0.01), higher respiratory rates ((27.3±2.5) times/min vs. (21.5±4.0) times/min, P<0.01), less 24 hours urine output ((422±266) ml vs. (1 680±863) ml, P<0.01), lower platelet counts ((161.9±81.9)×10⁹/L vs. (241.6±94.0)×10⁹/L, P=0.03), higher serum creatinine ((250.0±36.8) μmol/L vs. (132.7±34.2) μmol/L, P<0.01), higher alanine aminotransferase (288(76,846) IU/ml vs. 81(42, 109) IU/ml, P=0.04), lower artery pH (7.11±0.17 vs. 7.39±0.09, P<0.01), higher lactic acid ((10.29±3.62) mmol/L vs. (4.21±2.85) mmol/L, P<0.01), higher incidence of invasive ventilation (7/13 vs. 2/11, P=0.02), higher scores of acute physiology and chronic health evaluation (APACHE) Ⅱ (35.4±6.8 vs. 18.7±1.7, P<0.01) and simplified acute physiology score (SAPS) Ⅱ (73.5±17.4 vs. 47.0±4.3, P<0.01), and higher incidence of target vessel TIMI flow grade 0-2 (10/13 vs. 1/11, P<0.01). Kaplan-Meier survival curve analysis showed that survival rate at 28 days post the diagnosis of shock in TIMI flow grade 3 group was higher than that in TIMI flow grade 0-2 group (76.9% vs. 9.1%, log-rank test, P<0.01), and mortality rate was similar at 28 days post the diagnosis of shock between non-three-vessel lesions group and three-vessel lesions group (35.7% vs. 60.0%, log-rank test, P=0.14). Multivariate logistic regression analysis showed that compared with TIMI flow grade 0-2 group, the OR value of death at 28 days post the diagnosis of shock in TIMI flow grade 3 patients with acute myocardial infarction complicating cardiogenic shock due to left main disease was 0.030(95%CI 0.003-0.340, P<0.01). Conclusion Short-term outcomeof patients with acute myocardial infarction complicating cardiogenic shock due to left main disease remains poor, and final flow of TIMI grade 3 is confirmed as independent protective factor of death at 28 days post the diagnosis of shock in these patients.

Objective To investigate the different expression and prognostic significance of forkhead box M1 ( FOXM1) and Gli?l in ovarian high grade serous carcinoma ( HGSC) . Methods The expressions of FOXM1 and Gli?1 in 94 cases of HGSC and 20 cases of normal fallopian tube tissues were detected by immunohistochemistry. Kaplan?Meier analysis and Cox multivariate survival analysis were used to assess the relationship of the FOXM1 and Gli?1 levels with age, International Federation of Gynecology and Obstetrics ( FIGO) stage, omental metastasis, and residual foci and prognosis of HGSC. Results The positive rates of FOXM1 and Gli?1 expression in HGSC were 79.8% (75/94) and 77.7% (73/94), respectively, both significantly higher than those of the normal controls ( P<0.05) . The expressions of FOXM1 and Gli?1 were significantly correlated with FIGO stage, and both of their positive rates in stage Ⅲ-Ⅳpatients were significantly higher than those in stage Ⅰ-Ⅱ cases ( P<0.001) . The expressions of FOXM1 in HGSC were positively correlated with Gli?1. Kaplan?Meier analysis revealed that the 5?year overall survival rates of FOXM1?and Gli?1?positive groups were 8.0% and 6.8%, significantly lower than 36.8% and 38.1% of the FOXM1?and Gli?1?negative groups, respectively ( P<0. 05 for both ) . Cox multivariate survival analysis revealed that FIGO stage and overexpression of FOXM1 protein were independent prognostic factors of HGSC patients ( P<0.05 for both) . Conclusions The overexpression of FOXM1 and Gli?1 proteins participate in the carcinogenesis of HGSC, and are significantly associated with FIGO stage. The protein expression of FOXM1 is positively correlated with Gli?1 in HGSC. Expression of FOXM1 protein and FIGO stage are independent prognostic factors of HGSC.

Objective To investigate the different expression and prognostic significance of forkhead box M1 ( FOXM1) and Gli?l in ovarian high grade serous carcinoma ( HGSC) . Methods The expressions of FOXM1 and Gli?1 in 94 cases of HGSC and 20 cases of normal fallopian tube tissues were detected by immunohistochemistry. Kaplan?Meier analysis and Cox multivariate survival analysis were used to assess the relationship of the FOXM1 and Gli?1 levels with age, International Federation of Gynecology and Obstetrics ( FIGO) stage, omental metastasis, and residual foci and prognosis of HGSC. Results The positive rates of FOXM1 and Gli?1 expression in HGSC were 79.8% (75/94) and 77.7% (73/94), respectively, both significantly higher than those of the normal controls ( P<0.05) . The expressions of FOXM1 and Gli?1 were significantly correlated with FIGO stage, and both of their positive rates in stage Ⅲ-Ⅳpatients were significantly higher than those in stage Ⅰ-Ⅱ cases ( P<0.001) . The expressions of FOXM1 in HGSC were positively correlated with Gli?1. Kaplan?Meier analysis revealed that the 5?year overall survival rates of FOXM1?and Gli?1?positive groups were 8.0% and 6.8%, significantly lower than 36.8% and 38.1% of the FOXM1?and Gli?1?negative groups, respectively ( P<0. 05 for both ) . Cox multivariate survival analysis revealed that FIGO stage and overexpression of FOXM1 protein were independent prognostic factors of HGSC patients ( P<0.05 for both) . Conclusions The overexpression of FOXM1 and Gli?1 proteins participate in the carcinogenesis of HGSC, and are significantly associated with FIGO stage. The protein expression of FOXM1 is positively correlated with Gli?1 in HGSC. Expression of FOXM1 protein and FIGO stage are independent prognostic factors of HGSC.