Objective:To establish a day 4 embryo evaluation protocol by analyzing embryonic characteristics affecting blastocyst formation, and validate its clinical effectiveness.Methods:This retrospective cohort study included clinical data from 1 037 patients who underwent fresh in vitro fertilization and embryo transfer (IVF-ET) on day 4 in Center for Reproductive Medicine of Xingtai Meihe Reproductive and Genetic Hospital between January 2018 and April 2024. Morphological assessments were performed at (92±2) h post-fertilization. After excluding 1 326 embryos selected for fresh transfer, 2 723 embryos underwent blastocyst culture. To address selection bias in transferred embryos, the scoring system was primarily based on high-quality blastocyst formation rates. Multivariate binary regression analysis evaluated how day 4 developmental stage, fragmentation rate, stage-specific cleavage patterns, multinucleation/vacuolization affected transferable blastocyst formation rate and high-quality blastocyst rate. Regression coefficients determined parameter weights for high-quality blastocyst formation, establishing a day 4 embryo scoring protocol that compared outcomes across different grades. The scoring system was validated by comparing transferable blastocyst formation rate, high-quality blastocyst rate, and implantation rate among different day 4 embryo grades. All embryos were further rescored according to three previously reported evaluation schemes [Feil 2008, Gemma 2015, and European Society of Human Reproduction and Embryology (ESHRE) 2011]. The predictive values of these three day 4 scoring systems and the day 4 scoring system established in this study were compared using the area under the curve (AUC) receiver operating characteristic (ROC) curve in predicting the formation rate of transferable blastocyst, the formation rate of high-quality blastocyst, and implantation rate. Results:In the prediction of high-quality blastocyst formation, early blastocyst showed the highest influence weight compared to embryos with the ratio of blastomere numbers on day 4 to those on day 3 (BNR) <1.2 ( B=3.398, OR=29.915, P<0.001), followed by fragmentation <10% versus ≥50% ( B=1.263, OR=3.535, P<0.001), a stage-specific cleavage pattern ( B=0.903, OR=2.467, P=0.005), and absence of multinucleation or vacuoles ( B=0.797, OR=2.218, P=0.007). Using the newly developed day 4 scoring system, embryos were graded A, B, C, D, E. Transferable blastocyst formation rates were 88.57% (279/315), 76.99% (241/313), 56.11% (280/499), 40.27% (238/591) and 14.22% (143/1 005), respectively; high-quality blastocyst rates were 51.42% (162/315), 35.46% (111/313), 20.04% (100/499), 9.47% (56/591) and 3.98% (40/1 005). All inter-group differences were statistically significant (all P<0.005). Implantation rates for transferred embryos of grades A-E declined sequentially: 63.18% (381/603), 56.19% (322/573), 38.29% (54/141), 26.53% (13/49) and 9.67% (3/31). The day 4 embryo scoring system proposed in this study demonstrated significantly higher predictive efficacy for transferable blastocyst formation rate (AUC=0.812), high-quality blastocyst formation rate (AUC=0.804), and implantation rate (AUC=0.603) compared with Feil 2008 (AUC=0.797, P<0.001; AUC=0.781, P<0.001; AUC=0.585, P<0.001), Gemma 2015 (AUC=0.773, P<0.001; AUC=0.771, P<0.001; AUC=0.542, P=0.006), and ESHRE 2011 (AUC=0.710, P<0.001; AUC=0.745, P<0.001; AUC=0.592, P<0.001). We also observed the presence of pseudo-compacted embryo, whose transferable blastocyst formation rate [38.28% (49/128)] and high-quality blastocyst formation rate [7.03% (9/128)] were similar to those of cleavage-stage embryos with a BNR≥1.2. Conclusion:The established day 4 morphological assessment system reliably predicts the potential to transferable blastocyst and high-quality blastocyst, and effectively forecasts implantation rates.

Objective:To establish a day 4 embryo evaluation protocol by analyzing embryonic characteristics affecting blastocyst formation, and validate its clinical effectiveness.Methods:This retrospective cohort study included clinical data from 1 037 patients who underwent fresh in vitro fertilization and embryo transfer (IVF-ET) on day 4 in Center for Reproductive Medicine of Xingtai Meihe Reproductive and Genetic Hospital between January 2018 and April 2024. Morphological assessments were performed at (92±2) h post-fertilization. After excluding 1 326 embryos selected for fresh transfer, 2 723 embryos underwent blastocyst culture. To address selection bias in transferred embryos, the scoring system was primarily based on high-quality blastocyst formation rates. Multivariate binary regression analysis evaluated how day 4 developmental stage, fragmentation rate, stage-specific cleavage patterns, multinucleation/vacuolization affected transferable blastocyst formation rate and high-quality blastocyst rate. Regression coefficients determined parameter weights for high-quality blastocyst formation, establishing a day 4 embryo scoring protocol that compared outcomes across different grades. The scoring system was validated by comparing transferable blastocyst formation rate, high-quality blastocyst rate, and implantation rate among different day 4 embryo grades. All embryos were further rescored according to three previously reported evaluation schemes [Feil 2008, Gemma 2015, and European Society of Human Reproduction and Embryology (ESHRE) 2011]. The predictive values of these three day 4 scoring systems and the day 4 scoring system established in this study were compared using the area under the curve (AUC) receiver operating characteristic (ROC) curve in predicting the formation rate of transferable blastocyst, the formation rate of high-quality blastocyst, and implantation rate. Results:In the prediction of high-quality blastocyst formation, early blastocyst showed the highest influence weight compared to embryos with the ratio of blastomere numbers on day 4 to those on day 3 (BNR) <1.2 ( B=3.398, OR=29.915, P<0.001), followed by fragmentation <10% versus ≥50% ( B=1.263, OR=3.535, P<0.001), a stage-specific cleavage pattern ( B=0.903, OR=2.467, P=0.005), and absence of multinucleation or vacuoles ( B=0.797, OR=2.218, P=0.007). Using the newly developed day 4 scoring system, embryos were graded A, B, C, D, E. Transferable blastocyst formation rates were 88.57% (279/315), 76.99% (241/313), 56.11% (280/499), 40.27% (238/591) and 14.22% (143/1 005), respectively; high-quality blastocyst rates were 51.42% (162/315), 35.46% (111/313), 20.04% (100/499), 9.47% (56/591) and 3.98% (40/1 005). All inter-group differences were statistically significant (all P<0.005). Implantation rates for transferred embryos of grades A-E declined sequentially: 63.18% (381/603), 56.19% (322/573), 38.29% (54/141), 26.53% (13/49) and 9.67% (3/31). The day 4 embryo scoring system proposed in this study demonstrated significantly higher predictive efficacy for transferable blastocyst formation rate (AUC=0.812), high-quality blastocyst formation rate (AUC=0.804), and implantation rate (AUC=0.603) compared with Feil 2008 (AUC=0.797, P<0.001; AUC=0.781, P<0.001; AUC=0.585, P<0.001), Gemma 2015 (AUC=0.773, P<0.001; AUC=0.771, P<0.001; AUC=0.542, P=0.006), and ESHRE 2011 (AUC=0.710, P<0.001; AUC=0.745, P<0.001; AUC=0.592, P<0.001). We also observed the presence of pseudo-compacted embryo, whose transferable blastocyst formation rate [38.28% (49/128)] and high-quality blastocyst formation rate [7.03% (9/128)] were similar to those of cleavage-stage embryos with a BNR≥1.2. Conclusion:The established day 4 morphological assessment system reliably predicts the potential to transferable blastocyst and high-quality blastocyst, and effectively forecasts implantation rates.

ObjectiveTo observe the effects of Baihe Yuzi prescription （BYP） on the cystic fibrosis transmembrane conductance regulator （CFTR）， aquaporin （AQP）， zinc/iron-regulated transporter-like protein （ZIP） and local oxidative stress in epididymis of oligoasthenozoospermia （OAS） rats， and to explore the mechanism of its intervention in OAS. MethodAfter 35 rats were acclimatized for 1 week， 7 rats were randomly selected as the normal group， and the remaining 28 rats were given tripterygium glycosides （TG） 30 mg·kg^-1. After 4 weeks of modeling， they were randomly divided into 4 groups： model group， BYP low-dose group （LBYP）， BYP high-dose group （HBYP） and levocarnitine group， with 7 rats in each group. The rats in the normal group and model group were given normal saline at the same dosage. The levocarnitine group rats were given L-carnitine oral liquid （100 mg·kg^-1） by gavage. The LBYP group rats were given BYP 6.3 g·kg^-1， and the HBYP group rats were given BYP 12.6 g·kg^-1 by gavage once a day for consecutive 4 weeks. After the end of the intervention， sperm count and motility of all rats were detected， the histopathological structure of epididymis was observed by hematoxylin-eosin （HE） staining， and the expressions of CFTR， AQP9， AQP3， ZIP8， ZIP12 and other proteins were detected by Western blot. The contents of α-glycosidase （α-GC）， sialic acid （SA）， carnitine， superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px） and malondialdehyde （MDA） were detected by enzyme-linked immunosorbent assay （ELISA）. Total zinc content was measured using an inductively coupled plasma mass spectrometer. Free zinc ion content was detected by zinc ion probes. ResultCompared with those in the normal group， the sperm count and motility of rats were decreased and the epididymal structure was disordered in the model group. The contents of α-GC and carnitine were decreased in epididymis （P<0.05）. MDA levels were increased， while SOD， GSH-Px and zinc levels were decreased （P<0.05）. The expressions of CFTR and ZIP12 in the head and cauda of the epididymis were down-regulated， and AQP3 expression was up-regulated. The expression of ZIP8 in the cauda epididymis was up-regulated （P<0.05）. Compared with the model group， BYP can significantly improve the sperm count and motility， the epididymal structure of OAS rats and the levels of α-GC and carnitine （P<0.05）. The expressions of CFTR and ZIP12 in the head and cauda of the epididymis were up-regulated， while the expressions of ZIP8 in the cauda epididymis and AQP3 in the head of the epididymis were decreased （P<0.05）. The SOD and GSH-Px levels and total zinc content in epididymis were increased， and the MDA levels were decreased （P<0.05）. ConclusionBYP may improve the sperm quality and repair epididymal tissue structure and function of OAS rats， by regulating the expressions of CFTR， AQP3， and ZIP12 ion channels and local antioxidant mechanism.

Objective: To investigate the mediating effects of resilience and emotion regulation on family cumulative risk and positive coping style among primary and middle school students, so as to provide insights into promoting psychological health among them. Methods: The primary and middle school students from three counties (districts) of Baise City, Guangxi Zhuang Autonomous Region were selected through the stratified cluster random sampling method. Resilience, emotion regulation, family cumulative risk and positive coping style were investigated using the Resilience Scale for Chinese adolescent, Emotion Regulation Questionnaire, Chinese family Assessment Instrument, and Simple Coping Strategies Questionnaire, respectively. Mediating effects of resilience and emotion regulation on family cumulative risk and positive coping style were examined using Process macro program. Results: Totally 1 076 questionnaires were allocated, and 1 006 valid questionnaires were recovered, with an effective rate of 93.49%. There were 515 boys, accounting for 51.19%. The median age was 13 (interquartile range, 1) years. The mean scores of resilience, emotion regulation and positive coping style were (3.36±0.63), (2.26±0.56) and (1.85±0.39) points, respectively. The median family cumulative risk index was 0 (interquartile range, 0). The results of mediating effects showed that family cumulative risk had a direct negative effect on positive coping style (β=-0.037, 95%CI: -0.059 to -0.014, P=0.011), and also had an indirect negative effect on positive coping style through psychological resilience (β=-0.029, 95%CI: -0.041 to -0.017, P=0.006) as a mediator or through psychological resilience and emotion regulation (β=-0.002, 95%CI: -0.004 to -0.001, P=0.001) as a chain mediator. The mediating effect of resilience contributed 44% to the total effect, while the chain mediating effect of resilience and emotion regulation contributed 3%. Conclusion Resilience and emotional regulation serve as negative mediators between family cumulative risk and positive coping style among middle and primary school students.

Zinc is an important regulator of various biological processes such as protein synthesis,cell proliferation,migration and autophagy.The zinc transporter ZIP13(SLC39A13)is a homodimeric metal transporter functioning in transport of zinc,iron,and calcium plasma in body.ZIP13 is involved in disease by regulating zinc homeostasis,such as the key role of ZIP13 in connective tissue development because it is involved in the BMP/TGF-β signaling pathway.ZIP13 promotes the metastasis of human ovarian cancer cells by activating the Src/FAK signaling pathway and ZIP13 inhibits the biogenesis and energy expenditure of beige adipocytes by regulating C/EBP-β expression.

Objective:To explore the effect of different transplantation schemes on the clinical outcomes and maternal and fetal outcomes of fresh cycle on day 4.Methods:The clinical data of 745 cycles of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in Reproductive Medicine Center of Xingtai Infertility Specialist Hospitals from January 2019 to December 2021 were analyzed retrospectively. In all cycles, 1-2 embryos were transferred on day 4 after fertilization. According to different embryo transfer schemes, they were divided into five groups: one partial compaction embryo group (group A, n=17), two partial compaction embryos group (group B, n=236), one full compaction above embryo group (group C, n=241), partial compaction+full compaction above group (group D, n=72), and two full compaction above embryos group (group E, n=179), the effects of different transfer schemes on clinical outcomes and maternal and fetal outcomes were compared. Results:There were no significant differences in two pronuclei (2PN) fertilization rate, 2PN cleavage rate, endometrial thickness, monozygotic twins rate, surgical reduction rate, abortion rate, stillbirth rate, cesarean section rate, male infant/female infant ratio among the five groups (all P>0.05). The clinical pregnancy rate in groups D and E [83.33% (60/72), 72.63% (130/179)] was significantly higher than that in group B [59.32% (140/236), P<0.001, P=0.005]. The live birth rate of group E [65.36% (117/179)] was significantly higher than that of group B [50.00% (118/236), P=0.002]. The implantation rate of groups C, D and E [64.73% (156/241),59.72% (86/144), 54.19% (194/358)] was significantly higher than that of group B [39.83% (188/472), all P<0.001]. The multiple birth rate [43.33% (26/60), 49.23% (64/130)] and the premature delivery rate [23.33% (14/60), 26.15% (34/130)] in groups D and E were significantly higher than those in group C [0%, all P<0.001; 7.69% (12/156), P=0.002, P<0.001]. The gestational weeks of delivery in group E [38.0 (36.0,39.0) weeks] was significantly lower than that in group C [39.0 (38.0,39.3) weeks, P<0.001]. Compared with the one embryo transfer group (group A, group C), the birth weight of the two embryos transfer group (group B, group D, group E) was significantly lower ( P<0.001). The low birth weight infants rate in group D [20.90% (14/67)] and group E [28.40% (48/169)] was significantly higher than that in group C [3.85% (5/130), P<0.001]. Logistic regression analysis showed that after adjusted, the pregnancy rate in group C was significantly higher than that in group A ( OR=0.353, 95% CI: 0.125-0.997, P=0.049). The live birth rate of group E was significantly higher than that of group C ( OR=1.842, 95% CI: 1.172-2.895, P=0.008), and the premature delivery rate of groups B, D and E was significantly higher than that of group C ( OR=2.546, 95% CI: 1.079-6.005, P=0.033; OR=6.737, 95% CI: 2.566-17.689, P<0.001; OR=5.863, 95% CI: 2.494-13.785, P<0.001). Conclusion:On day 4 of the fresh cycle, the pregnancy rate and the live birth rate of the full compaction or above embryos are significantly higher than those of the partial compaction embryos. In clinical work, it is recommended to select full compaction embryos or above, and then select partially compaction embryos for single embryo transfer to achieve optimal pregnancy outcomes while reducing the multiple pregnancy rate, premature delivery rate, and low birth weight infant rate.

Objective:To investigate the effects of day 4 (D4) embryo transfer on clinical outcomes and maternal and infant outcomes in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in fresh cycles. Methods:We retrospectively analyzed the clinical data of 361 IVF and ICSI cycles from January 2019 to December 2021 in the Reproductive Medicine Center of Xingtai Infertility Specialist Hospitals in a cohort study. In all cycles, one of the best quality embryo was transferred at D4 or D5 after fertilization. The clinical outcomes and maternal and fetal outcomes of D4 transfer group and D5 transfer group were compared.Results:Compared with D4 transfer group, the estradiol value on human chorionic gonadotropin (hCG) injection day significantly increased in D5 transfer group [3 516 (2 273, 4 116) ng/L vs. 2 940 (1 935, 3 877) ng/L, P=0.023], and the luteinizing hormone (LH) value on hCG injection day significantly decreased [0.82 (0.63, 1.11) μg/L vs. 0.94 (0.69, 1.20) μg/L, P=0.026], two pronuclei (2PN) fertilization rate [71.13% (929/1 306)] and the available embryo rate [51.73% (509/984)] in D5 transfer group were significantly higher than those in D4 transfer group [67.26% (2 539/3 775), P=0.010; 48.00% (1 306/2 721), P=0.045]. The clinical pregnancy rate [63.37% (173/273)] and the live birth rate [53.11% (145/273)] in D4 transfer group were significantly higher than those in D5 transfer group [46.59% (41/88), P=0.005; 36.36% (32/88), P=0.006]. There were no significant differences in ectopic pregnancy rate, monozygotic twins rate, abortion rate, preterm birth rate, stillbirth rate, caesarean section rate, delivery gestation, male/female ratio and birth weight (all P>0.05). The results of logistic regression analysis showed that the days of embryo transfer had a significant correlation with the pregnancy rate and the live birth rate ( OR=0.468, 95% CI: 0.266-0.824, P=0.009; OR=0.481, 95% CI: 0.273-0.847, P=0.011). Conclusion:In IVF/ICSI of fresh cycles, the clinical pregnancy rate and the live birth rate of D4 transplantation are better than that of D5 transplantation, and the maternal and fetal outcomes are similar. Therefore, D4 embryo transplantation can be preferred in clinical work.

Objective:To explore the effect of different transplantation schemes on the clinical outcomes and maternal and fetal outcomes of fresh cycle on day 4.Methods:The clinical data of 745 cycles of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in Reproductive Medicine Center of Xingtai Infertility Specialist Hospitals from January 2019 to December 2021 were analyzed retrospectively. In all cycles, 1-2 embryos were transferred on day 4 after fertilization. According to different embryo transfer schemes, they were divided into five groups: one partial compaction embryo group (group A, n=17), two partial compaction embryos group (group B, n=236), one full compaction above embryo group (group C, n=241), partial compaction+full compaction above group (group D, n=72), and two full compaction above embryos group (group E, n=179), the effects of different transfer schemes on clinical outcomes and maternal and fetal outcomes were compared. Results:There were no significant differences in two pronuclei (2PN) fertilization rate, 2PN cleavage rate, endometrial thickness, monozygotic twins rate, surgical reduction rate, abortion rate, stillbirth rate, cesarean section rate, male infant/female infant ratio among the five groups (all P>0.05). The clinical pregnancy rate in groups D and E [83.33% (60/72), 72.63% (130/179)] was significantly higher than that in group B [59.32% (140/236), P<0.001, P=0.005]. The live birth rate of group E [65.36% (117/179)] was significantly higher than that of group B [50.00% (118/236), P=0.002]. The implantation rate of groups C, D and E [64.73% (156/241),59.72% (86/144), 54.19% (194/358)] was significantly higher than that of group B [39.83% (188/472), all P<0.001]. The multiple birth rate [43.33% (26/60), 49.23% (64/130)] and the premature delivery rate [23.33% (14/60), 26.15% (34/130)] in groups D and E were significantly higher than those in group C [0%, all P<0.001; 7.69% (12/156), P=0.002, P<0.001]. The gestational weeks of delivery in group E [38.0 (36.0,39.0) weeks] was significantly lower than that in group C [39.0 (38.0,39.3) weeks, P<0.001]. Compared with the one embryo transfer group (group A, group C), the birth weight of the two embryos transfer group (group B, group D, group E) was significantly lower ( P<0.001). The low birth weight infants rate in group D [20.90% (14/67)] and group E [28.40% (48/169)] was significantly higher than that in group C [3.85% (5/130), P<0.001]. Logistic regression analysis showed that after adjusted, the pregnancy rate in group C was significantly higher than that in group A ( OR=0.353, 95% CI: 0.125-0.997, P=0.049). The live birth rate of group E was significantly higher than that of group C ( OR=1.842, 95% CI: 1.172-2.895, P=0.008), and the premature delivery rate of groups B, D and E was significantly higher than that of group C ( OR=2.546, 95% CI: 1.079-6.005, P=0.033; OR=6.737, 95% CI: 2.566-17.689, P<0.001; OR=5.863, 95% CI: 2.494-13.785, P<0.001). Conclusion:On day 4 of the fresh cycle, the pregnancy rate and the live birth rate of the full compaction or above embryos are significantly higher than those of the partial compaction embryos. In clinical work, it is recommended to select full compaction embryos or above, and then select partially compaction embryos for single embryo transfer to achieve optimal pregnancy outcomes while reducing the multiple pregnancy rate, premature delivery rate, and low birth weight infant rate.

Objective:To investigate the effects of day 4 (D4) embryo transfer on clinical outcomes and maternal and infant outcomes in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in fresh cycles. Methods:We retrospectively analyzed the clinical data of 361 IVF and ICSI cycles from January 2019 to December 2021 in the Reproductive Medicine Center of Xingtai Infertility Specialist Hospitals in a cohort study. In all cycles, one of the best quality embryo was transferred at D4 or D5 after fertilization. The clinical outcomes and maternal and fetal outcomes of D4 transfer group and D5 transfer group were compared.Results:Compared with D4 transfer group, the estradiol value on human chorionic gonadotropin (hCG) injection day significantly increased in D5 transfer group [3 516 (2 273, 4 116) ng/L vs. 2 940 (1 935, 3 877) ng/L, P=0.023], and the luteinizing hormone (LH) value on hCG injection day significantly decreased [0.82 (0.63, 1.11) μg/L vs. 0.94 (0.69, 1.20) μg/L, P=0.026], two pronuclei (2PN) fertilization rate [71.13% (929/1 306)] and the available embryo rate [51.73% (509/984)] in D5 transfer group were significantly higher than those in D4 transfer group [67.26% (2 539/3 775), P=0.010; 48.00% (1 306/2 721), P=0.045]. The clinical pregnancy rate [63.37% (173/273)] and the live birth rate [53.11% (145/273)] in D4 transfer group were significantly higher than those in D5 transfer group [46.59% (41/88), P=0.005; 36.36% (32/88), P=0.006]. There were no significant differences in ectopic pregnancy rate, monozygotic twins rate, abortion rate, preterm birth rate, stillbirth rate, caesarean section rate, delivery gestation, male/female ratio and birth weight (all P>0.05). The results of logistic regression analysis showed that the days of embryo transfer had a significant correlation with the pregnancy rate and the live birth rate ( OR=0.468, 95% CI: 0.266-0.824, P=0.009; OR=0.481, 95% CI: 0.273-0.847, P=0.011). Conclusion:In IVF/ICSI of fresh cycles, the clinical pregnancy rate and the live birth rate of D4 transplantation are better than that of D5 transplantation, and the maternal and fetal outcomes are similar. Therefore, D4 embryo transplantation can be preferred in clinical work.

Objective To explore the value of contrast-enhanced ultrasound ( CEUS) in evaluating the effect of liver cancer ablation. Methods A total of 46 patients with liver cancer (60 lesions) were treated with microwave ablation between May 2008 and October 2016.After 1 month, color Doppler flow imaging ( CDFI), contrast-enhanced CT ( CECT) and CEUS examinations were performed.A biopsy was performed on each of positive lesions . Results A total of 12 positive lesions were detected by CECT , and the biopsy proved to be residual tumor .A total of 11 lesions were detected by CEUS (the rate of missed diagnosis was 8.3%, 1/12). In CEUS positive lesions, 8 lesions were detected by CDFI (the rate of missed diagnosis was 33.3%, 4/12).There were no false positives. Conclusions Application of CEUS to evaluate the effect of liver tumor ablation is simple , reliable, and non-radiative.It has obvious advantages in follow-ups after liver tumor ablation and guided biopsy and treatment of residual lesions .