ObjectiveTo explore the relationship between negative life events and depression severity in adolescent patients with depressive disorder， as well as the chain mediating role of insomnia symptoms and quality of life. Methods374 outpatient patients and hospitalized patients with adolescent depressive disorders were enrolled. The Adolescent Life Event Scale （ASLEC）， the Insomnia Severity Index （ISI）， the World Health Organization Quality of Life Questionnaire Short Form （WHOQOL-BREF）， and the Center for Epidemiology Depression Scale （CES-D） were used to evaluate the negative life event situation， insomnia symptoms， quality of life level and depression severity of the subjects， respectively. In addition， the PROCESS 4.0 macroprogram was used to analyze the chain mediating effect of insomnia symptoms and quality of life between negative life events and depression severity in patients with adolescent depressive disorder. ResultsThe results of correlation analysis showed that there was a significant correlation between negative life events and insomnia symptoms， quality of life， and depression severity （all P<0.05）. In addition， the results of chain mediation showed that negative life events had a significant direct effect on depression severity， with an effect size of 0.12 （P<0.001）. Insomnia symptoms and quality of life played a mediating role in the relationship between negative life events and depression severity in patients with adolescent depressive disorders， with indirect effect sizes of 0.062 （95%CI： 0.040-0.087） and 0.091 （95%CI： 0.059-0.123）， respectively. It could also play a chain mediation role， and the effect size was 0.039 （95%CI： 0.024-0.057）. ConclusionNegative life events experienced by patients with adolescent depressive disorder not only directly affect the severity of depressive symptoms， but may also indirectly exacerbate depression through insomnia symptoms and quality of life.

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.

BACKGROUND:The extracellular-regulated protein kinase 5(ERK5)signaling protein is essential for the survival of organisms,and resveratrol can promote osteoblast proliferation through various pathways.However,whether resveratrol can regulate osteoblast function through the ERK5 signaling protein needs further verification. OBJECTIVE:To explore the regulatory effect of ERK5 on the proliferation of MC3T3-E1 cells and related secreted proteins,and to further verify whether resveratrol can complete the above process by activating ERK5. METHODS:Mouse MC3T3-E1 preosteoblasts were treated with complete culture medium,XMD8-92(an ERK5 inhibitor),epidermal growth factor(an ERK5 activator),resveratrol alone,XMD8-92+EGF,and resveratrol+XMD8-92,respectively.Western blot assay was used to detect the expression of ERK5 and p-ERK5 proteins,proliferation-related proteins Cyclin D1,CDK4 and PCNA,and osteoblast-secreted proteins osteoprotegerin and receptor activator of nuclear factor-κB ligand in MC3T3-E1 cells of each group.The fluorescence intensity of ERK5,osteoprotegerin and receptor activator of nuclear factor-κB ligand in each group was detected by cell immunofluorescence staining,and cell proliferation was detected by EdU staining,respectively.The appropriate concentration and time of resveratrol intervention in MC3T3-E1 cells were determined by cell morphology observation and cell counting kit-8 assay. RESULTS AND CONCLUSION:The activation of ERK5 signaling protein could effectively promote the proliferation of MC3T3-E1 cells,up-regulate the osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio.The appropriate concentration and time for resveratrol intervention in MC3T3-E1 cells was 5 μmol/L and 24 hours,respectively.Resveratrol could activate ERK5 signaling protein,thereby promoting osteoblast proliferation and up-regulating the osteoprotegerin/RANKL ratio.All these results indicate that resveratrol can promote the proliferation of MC3T3-E1 cells and up-regulate the osteoprotegerin/RANKL ratio by activating the ERK5 signaling protein.

Objective:To explore the clinical and pathological features and survival outcomes of patients with early-onset intrahepatic cholangiocarcinoma (EOICC).Methods:This is a multicenter, retrospective cohort study. Data of 1 160 intrahepatic cholangiocarcinoma patients undergoing radical resection in 14 tertiary Grade A hospitals in China from January 2010 to November 2021 were retrospectively collected. The cohort included 632 males and 528 females, aged( M (IQR)) 61 (14) years (range: 22 to 93 years). ICC aged ≤50 years at the time of diagnosis was defined as EOICC and >50 years as late-onset intrahepatic cholangiocarcinoma (LOICC). Of these, there were 247 cases in the EOICC group and 913 cases in the LOICC. The clinical and pathological characteristics of both groups were analyzed and compared using the independent sample t-test, Mann-Whitney U test or Kaplan-Meier method. Univariate and multivariate Cox regression models for patient outcomes were constructed and forest graphed. Results:Compared with the patients in the LOICC group, patients in the EOICC group had lower carcinoembryonic antigen levels (2.5(4.0) μg/L vs. 3.1(5.2)μg/L, U=124 899, P=0.009) and CA19-9 level (63.4(524.7)U/ml vs. 77.9(611.3)U/ml, U=120 320, P=0.013), higher levels of ALT (29(35)U/L vs. 24(26)U/L, U=101 214, P=0.013), a lower score of the Eastern US Cooperative Oncology Group (0 score patients: 54.7% vs. 44.1%, χ2=12.472, P=0.014), higher TNM stage ( χ2=11.807, P=0.038), and proportion of lymph node dissection (62.3% vs. 54.1%, χ2=5.355, P=0.021). Patients in the two groups in sex, first diagnosis symptoms, intrahepatic bile duct stone history, nail protein, albumin, total bilirubin, transaminase, liver function Child-Pugh grade, T stage, stage, N stage, preoperative laparoscopic exploration proportion, tumor diameter, vascular invasion proportion, differentiation, margin, intraoperative bleeding, postoperative complications, postoperative hospital days were no statistical significance (all P>0.05). Patients in the EOICC group had better outcomes than the LOICC group (median survival time: 29.7 months vs. 25.0 months, 3-year overall survival: 45.1% vs. 37.8%, P=0.027). Conclusion:EOICC patients are better than LOICC patients in carcinoembryonic antigen, CA19-9, ALT, physical strength status and TNM stage, and the long-term prognosis is also better than LOICC patients.

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 explore the clinical and pathological features and survival outcomes of patients with early-onset intrahepatic cholangiocarcinoma (EOICC).Methods:This is a multicenter, retrospective cohort study. Data of 1 160 intrahepatic cholangiocarcinoma patients undergoing radical resection in 14 tertiary Grade A hospitals in China from January 2010 to November 2021 were retrospectively collected. The cohort included 632 males and 528 females, aged( M (IQR)) 61 (14) years (range: 22 to 93 years). ICC aged ≤50 years at the time of diagnosis was defined as EOICC and >50 years as late-onset intrahepatic cholangiocarcinoma (LOICC). Of these, there were 247 cases in the EOICC group and 913 cases in the LOICC. The clinical and pathological characteristics of both groups were analyzed and compared using the independent sample t-test, Mann-Whitney U test or Kaplan-Meier method. Univariate and multivariate Cox regression models for patient outcomes were constructed and forest graphed. Results:Compared with the patients in the LOICC group, patients in the EOICC group had lower carcinoembryonic antigen levels (2.5(4.0) μg/L vs. 3.1(5.2)μg/L, U=124 899, P=0.009) and CA19-9 level (63.4(524.7)U/ml vs. 77.9(611.3)U/ml, U=120 320, P=0.013), higher levels of ALT (29(35)U/L vs. 24(26)U/L, U=101 214, P=0.013), a lower score of the Eastern US Cooperative Oncology Group (0 score patients: 54.7% vs. 44.1%, χ2=12.472, P=0.014), higher TNM stage ( χ2=11.807, P=0.038), and proportion of lymph node dissection (62.3% vs. 54.1%, χ2=5.355, P=0.021). Patients in the two groups in sex, first diagnosis symptoms, intrahepatic bile duct stone history, nail protein, albumin, total bilirubin, transaminase, liver function Child-Pugh grade, T stage, stage, N stage, preoperative laparoscopic exploration proportion, tumor diameter, vascular invasion proportion, differentiation, margin, intraoperative bleeding, postoperative complications, postoperative hospital days were no statistical significance (all P>0.05). Patients in the EOICC group had better outcomes than the LOICC group (median survival time: 29.7 months vs. 25.0 months, 3-year overall survival: 45.1% vs. 37.8%, P=0.027). Conclusion:EOICC patients are better than LOICC patients in carcinoembryonic antigen, CA19-9, ALT, physical strength status and TNM stage, and the long-term prognosis is also better than LOICC patients.

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.