ObjectiveTo study the accumulation and absorption of dry matter and nutrients in Curcuma phaeocaulis from Sichuan， the origin of Curcumae Radix （tuber） and Curcumae Rhizoma （rhizome）， to explore the growth and development laws and nutrient demand characteristics of the medicinal species， and thus to lay a theoretical basis for rational fertilization. MethodThe plant growth indexes， dry matter accumulation， and nutrient content of C. phaeocaulis at different growth stages in Sichuan were measured and analyzed. ResultThis medicinal species featured the dominant growth of aboveground leaves and stems before October and growth of underground part （particularly the rhizomes and tubers） from October. During the whole growth period， the accumulation of nitrogen， phosphorus， potassium， calcium， magnesium， iron， manganese， zinc， and copper per plant was 2 450.31， 907.09， 3 171.18， 625.94， 493.38， 14.53， 2.24 ， 2.93， 0.46 mg， respectively， with the order of potassium > nitrogen > phosphorus > calcium > magnesium > iron > zinc > manganese > copper. ConclusionThe species needs sufficient potassium and nitrogen， appropriate amount of phosphorus， calcium， and magnesium， a small amount of iron， and very little zinc， manganese， and copper for growth， and potassium is particularly important. Nitrogen， phosphorus， and potassium fertilizers supply macroelements and nitrogen fertilizer should be supplemented at seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome. Phosphorus and potassium fertilizers should be applied at seedling stage， tuber expansion stage， secondary expansion stage of rhizome， and dry matter accumulation stage of tuber and rhizome. At seedling stage， leafy stage， early rhizome expansion stage， tuber expansion stage， and the second expansion stage of rhizome， a variety of medium and trace elements-containing foliar fertilizers should be used.

Objective:To investigate the effect of anti-Müllerian hormone (AMH) level on early pregnancy loss and cumulative pregnancy outcome of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI), and whether AMH level can be used as an indicator to evaluate the quality of oocytes. Methods:A retrospective cohort analysis was performed on 3701 women undergoing their first oocyte retrieval for IVF/ICSI at the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from December 2015 to December 2019. According to the age, patients were divided into elderly group (≥35 years old) and younger group (<35 years old). References to the Bologna criteria and POSEIDON criteria, patients were divided into three groups: low AMH group (AMH≤1 μg/L), moderately low AMH group (1 μg/L0.05) whether in the elderly group or the younger group. The cumulative pregnancy rates were 61.9% (211/341), 66.0% (319/483), 69.3% (1219/1760), and the cumulative live birth rates were 50.1% (171/341), 57.8% (279/483), 57.1% (1005/1760) in low AMH group, moderately low AMH group and normal AMH group of the younger group, respectively,and there were statistical differences ( P=0.020, P=0.047). After adjusting for confounding factors, in the younger group, cumulative pregnancy rate and cumulative live birth rate in the low AMH group were lower than those in the normal AMH group ( OR=0.617, 95% CI=0.460-0.829, P=0.001; OR=0.660, 95% CI=0.496-0.878, P=0.004), the cumulative pregnancy rate and the cumulative live birth rate in the moderately low AMH group were lower than those in the normal AMH group, but there was no statistical difference (all P>0.05). There were no significant differences in cumulative pregnancy rate and cumulative live birth rate among the three groups with different AMH levels in the elderly group (all P>0.05). Conclusion:AMH is not associated with the incidence of early pregnancy loss during the their first oocyte retrieval in IVF/ICSI. AMH level is an independent factor affecting the cumulative pregnancy outcome in the younger group, the pregnancy outcome of women with AMH ≤1 μg/L decreased significantly. In the eldly women, AMH was not significantly associated with cumulative pregnancy outcome. AMH level may not be an indicator of oocyte quality.

Objective:To investigate the effect of anti-Müllerian hormone (AMH) level on early pregnancy loss and cumulative pregnancy outcome of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI), and whether AMH level can be used as an indicator to evaluate the quality of oocytes. Methods:A retrospective cohort analysis was performed on 3701 women undergoing their first oocyte retrieval for IVF/ICSI at the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from December 2015 to December 2019. According to the age, patients were divided into elderly group (≥35 years old) and younger group (<35 years old). References to the Bologna criteria and POSEIDON criteria, patients were divided into three groups: low AMH group (AMH≤1 μg/L), moderately low AMH group (1 μg/L0.05) whether in the elderly group or the younger group. The cumulative pregnancy rates were 61.9% (211/341), 66.0% (319/483), 69.3% (1219/1760), and the cumulative live birth rates were 50.1% (171/341), 57.8% (279/483), 57.1% (1005/1760) in low AMH group, moderately low AMH group and normal AMH group of the younger group, respectively,and there were statistical differences ( P=0.020, P=0.047). After adjusting for confounding factors, in the younger group, cumulative pregnancy rate and cumulative live birth rate in the low AMH group were lower than those in the normal AMH group ( OR=0.617, 95% CI=0.460-0.829, P=0.001; OR=0.660, 95% CI=0.496-0.878, P=0.004), the cumulative pregnancy rate and the cumulative live birth rate in the moderately low AMH group were lower than those in the normal AMH group, but there was no statistical difference (all P>0.05). There were no significant differences in cumulative pregnancy rate and cumulative live birth rate among the three groups with different AMH levels in the elderly group (all P>0.05). Conclusion:AMH is not associated with the incidence of early pregnancy loss during the their first oocyte retrieval in IVF/ICSI. AMH level is an independent factor affecting the cumulative pregnancy outcome in the younger group, the pregnancy outcome of women with AMH ≤1 μg/L decreased significantly. In the eldly women, AMH was not significantly associated with cumulative pregnancy outcome. AMH level may not be an indicator of oocyte quality.

Objective:To investigate the effect of endometrial thickness on birth weight in frozen-thawed embryo transfer cycles.Methods:A retrospective cohort analysis of 1622 single-birth live birth cycles of frozen-thawed embryo transfer at the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from June 2016 to December 2018 was performed. According to the endometrial thickness on the day of transplantation, the patients were divided into four groups, group A: endometrial thickness ≤8 mm; group B: endometrial thickness 8-10 mm; group C: endometrial thickness 10-12 mm; group D: endometrial thickness ≥12 mm. The general and clinical data of patients in each group were compared. Group A was used as the reference group to control confounding factors and the low birth weight (LBW) and very low birth weight (VLBW), term LBW, small for gestational age (SGA), preterm birth odds ratio ( OR), and 95% confidence interval (95% CI) were calculated, and multiple linear regression was used to analyze the impact factors related to the birth weight of the fetus. Results:The fetal birth weights of the four groups were (3 350.15±568.17) g, (3 318.84±523.43) g, (3 459.93±483.73) g, (3 482.27±495.30) g in order ( P>0.05). After adjusting for confounding factors through multivariate logistic regression analysis, the risk of preterm birth in group A was greater than that in group C ( OR=0.527,95% CI=0.287-0.968, P=0.039). Endometrial thickness did not affect the incidence of LBW, VLBW, term LBW, and SGA ( P>0.05). Further, multiple linear regression analyses indicated that body mass index, embryo transfer stage, gestation week, and newborn gender all affected the fetal birth weight ( P<0.001, P=0.003, P<0.001, P<0.001). Conclusion:In the frozen-thawed embryo transfer cycle, the women with endometrial thickness ≤8 mm on the day of transplantation are more likely to have premature birth than women with endometrial thickness of 10-12 mm, but endometrial thickness does not affect the fetal birth weight.

Objective:To investigate the effect of endometrial thickness on birth weight in frozen-thawed embryo transfer cycles.Methods:A retrospective cohort analysis of 1622 single-birth live birth cycles of frozen-thawed embryo transfer at the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from June 2016 to December 2018 was performed. According to the endometrial thickness on the day of transplantation, the patients were divided into four groups, group A: endometrial thickness ≤8 mm; group B: endometrial thickness 8-10 mm; group C: endometrial thickness 10-12 mm; group D: endometrial thickness ≥12 mm. The general and clinical data of patients in each group were compared. Group A was used as the reference group to control confounding factors and the low birth weight (LBW) and very low birth weight (VLBW), term LBW, small for gestational age (SGA), preterm birth odds ratio ( OR), and 95% confidence interval (95% CI) were calculated, and multiple linear regression was used to analyze the impact factors related to the birth weight of the fetus. Results:The fetal birth weights of the four groups were (3 350.15±568.17) g, (3 318.84±523.43) g, (3 459.93±483.73) g, (3 482.27±495.30) g in order ( P>0.05). After adjusting for confounding factors through multivariate logistic regression analysis, the risk of preterm birth in group A was greater than that in group C ( OR=0.527,95% CI=0.287-0.968, P=0.039). Endometrial thickness did not affect the incidence of LBW, VLBW, term LBW, and SGA ( P>0.05). Further, multiple linear regression analyses indicated that body mass index, embryo transfer stage, gestation week, and newborn gender all affected the fetal birth weight ( P<0.001, P=0.003, P<0.001, P<0.001). Conclusion:In the frozen-thawed embryo transfer cycle, the women with endometrial thickness ≤8 mm on the day of transplantation are more likely to have premature birth than women with endometrial thickness of 10-12 mm, but endometrial thickness does not affect the fetal birth weight.

Objective:To investigate the effect of di-2-ethylhexyl phthalate (DEHP) on oocyte development potential and its mechanism.Methods:Forty 3-week old female KM mice were randomly divided into 4 groups ( n=10, each), including control group (corn oil), the low-, medium-, and high-dose DEHP groups (300 mg/kg, 600 mg/kg, 1200 mg/kg). Mice of 4 groups were orally administered with corn oil (control) or DEHP daily for 6 weeks (5 d per week), respectively. After the treatment, female mice in each group were given ovulation induction and cooped up with male mice. The fertilized eggs were collected to observe the cleavage rate and blastocyst formation rate. Meanwhile, the levels of Ca 2+ and reactive oxygen species (ROS) of oocytes in each group were detected with the Ca 2+ fluorescence probe (Fluo-4 AM) and ROS assay kit, then the differences were compared. Results:Compared with control group, the cleavage rate and blastocyst formation rate of the low-, medium- and high-dose DEHP groups decreased, and the differences were statistically significant ( P all<0.05). The levels of Ca 2+ and ROS in oocytes increased with the increase of DEHP dosage. Compared with control group and the low-dose DEHP group, the levels of Ca 2+ and ROS in oocytes of the medium- and high-dose DEHP groups were significantly higher, and the differences were statistically significant ( P all<0.05). Conclusion:DEHP can increase the levels of Ca 2+ and ROS in mouse oocytes, reduce the quality of oocytes, and affect the early embryonic development.

Objective:To investigate the impact of the proportion of follicle diameter ≥14 mm on human chorionic hormone (hCG) injection day and follicular output rate (FORT) on the pregnancy outcome of fresh embryo transfer in patients with polycystic ovary syndrome (PCOS) by using long-acting gonadotropin-releasing hormone agonist (GnRH-a) long protocol in follicular phase.Methods:The data of 811 fresh embryo transfer cycles of PCOS patients assisted by long-acting GnRH-a long protocol in follicular phase were analyzed retospectively from May 2017 to May 2018 in the Center for Reproductive Medicine of the Third Affiliated Hospital of Zhengzhou University. According to the proportion of follicles with a diameter ≥14 mm on hCG injection day, patients were divided into three groups: group A (proportion≤60%), group B (proportion 60%-80%) and group C (proportion≥80%). According to the FORT, patients were divided into three groups: group D (proportion≤40%), group E (proportion 40%-60%) and group F (proportion≥60%). The clinical and laboratory indicators of patients in each group were compared and analyzed. According to the pregnancy outcomes, patients were divided into pregnancy group and non-pregnancy group. The clinical data of the two groups were compared, and the influencing factors of clinical pregnancy outcomes were analyzed by binary logistic regression.Results:The number of retrieved oocytes of groups A, B and C increased in turn ( P<0.001). The fertility rate of two pronuclei (2PN) in group A (59.78%) was lower than that in group B (62.40%, P=0.013). There was no significant difference among the three groups in embryo implantation rate, clinical pregnancy rate and live birth rate ( P>0.05). The number of retrieved oocytes in groups D, E and F increased ( P<0.001), but the 2PN rate in group F (59.13%) was significantly lower than that in group D (62.98%, P=0.009) and group E (64.05%, P<0.001). Embryo implantation rate, clinical pregnancy rate and live birth rate in group F were lower than those in group E (51.76% vs. 61.56%, P=0.002; 62.72% vs. 72.70%, P=0.007; 52.33% vs. 61.84%, P=0.011), while those in group D (56.77%, 69.94%, 59.54%) were not significantly different from those in the other two groups ( P>0.05). Compared with pregnancy group, the proportion of dominant follicles on hCG injection day in non-pregnancy group was not significantly different ( P>0.05), while FORT in pregnancy group was lower than that in non-pregnancy group ( P=0.031). Logistic regression analysis showed that FORT was an independent influencing factor for clinical pregnancy [ OR=0.995, 95% CI=0.974-1.016, P=0.039]. Conclusion:In PCOS patients, the proportion of follicles with a diameter ≥14 mm on hCG injection day does not affect pregnancy outcomes, the FORT is of great significance in predicting pregnancy outcomes, and the middle and low proportion of FORT group can obtain better pregnancy outcomes.

Objective:To investigate the effect of di-2-ethylhexyl phthalate (DEHP) on oocyte development potential and its mechanism.Methods:Forty 3-week old female KM mice were randomly divided into 4 groups ( n=10, each), including control group (corn oil), the low-, medium-, and high-dose DEHP groups (300 mg/kg, 600 mg/kg, 1200 mg/kg). Mice of 4 groups were orally administered with corn oil (control) or DEHP daily for 6 weeks (5 d per week), respectively. After the treatment, female mice in each group were given ovulation induction and cooped up with male mice. The fertilized eggs were collected to observe the cleavage rate and blastocyst formation rate. Meanwhile, the levels of Ca 2+ and reactive oxygen species (ROS) of oocytes in each group were detected with the Ca 2+ fluorescence probe (Fluo-4 AM) and ROS assay kit, then the differences were compared. Results:Compared with control group, the cleavage rate and blastocyst formation rate of the low-, medium- and high-dose DEHP groups decreased, and the differences were statistically significant ( P all<0.05). The levels of Ca 2+ and ROS in oocytes increased with the increase of DEHP dosage. Compared with control group and the low-dose DEHP group, the levels of Ca 2+ and ROS in oocytes of the medium- and high-dose DEHP groups were significantly higher, and the differences were statistically significant ( P all<0.05). Conclusion:DEHP can increase the levels of Ca 2+ and ROS in mouse oocytes, reduce the quality of oocytes, and affect the early embryonic development.

Objective:To investigate the impact of the proportion of follicle diameter ≥14 mm on human chorionic hormone (hCG) injection day and follicular output rate (FORT) on the pregnancy outcome of fresh embryo transfer in patients with polycystic ovary syndrome (PCOS) by using long-acting gonadotropin-releasing hormone agonist (GnRH-a) long protocol in follicular phase.Methods:The data of 811 fresh embryo transfer cycles of PCOS patients assisted by long-acting GnRH-a long protocol in follicular phase were analyzed retospectively from May 2017 to May 2018 in the Center for Reproductive Medicine of the Third Affiliated Hospital of Zhengzhou University. According to the proportion of follicles with a diameter ≥14 mm on hCG injection day, patients were divided into three groups: group A (proportion≤60%), group B (proportion 60%-80%) and group C (proportion≥80%). According to the FORT, patients were divided into three groups: group D (proportion≤40%), group E (proportion 40%-60%) and group F (proportion≥60%). The clinical and laboratory indicators of patients in each group were compared and analyzed. According to the pregnancy outcomes, patients were divided into pregnancy group and non-pregnancy group. The clinical data of the two groups were compared, and the influencing factors of clinical pregnancy outcomes were analyzed by binary logistic regression.Results:The number of retrieved oocytes of groups A, B and C increased in turn ( P<0.001). The fertility rate of two pronuclei (2PN) in group A (59.78%) was lower than that in group B (62.40%, P=0.013). There was no significant difference among the three groups in embryo implantation rate, clinical pregnancy rate and live birth rate ( P>0.05). The number of retrieved oocytes in groups D, E and F increased ( P<0.001), but the 2PN rate in group F (59.13%) was significantly lower than that in group D (62.98%, P=0.009) and group E (64.05%, P<0.001). Embryo implantation rate, clinical pregnancy rate and live birth rate in group F were lower than those in group E (51.76% vs. 61.56%, P=0.002; 62.72% vs. 72.70%, P=0.007; 52.33% vs. 61.84%, P=0.011), while those in group D (56.77%, 69.94%, 59.54%) were not significantly different from those in the other two groups ( P>0.05). Compared with pregnancy group, the proportion of dominant follicles on hCG injection day in non-pregnancy group was not significantly different ( P>0.05), while FORT in pregnancy group was lower than that in non-pregnancy group ( P=0.031). Logistic regression analysis showed that FORT was an independent influencing factor for clinical pregnancy [ OR=0.995, 95% CI=0.974-1.016, P=0.039]. Conclusion:In PCOS patients, the proportion of follicles with a diameter ≥14 mm on hCG injection day does not affect pregnancy outcomes, the FORT is of great significance in predicting pregnancy outcomes, and the middle and low proportion of FORT group can obtain better pregnancy outcomes.