Objective:To analyze the clinical phenotype, sperm morphological characteristics and assisted reproductive therapy efficiency in patients with total globozoospermia.Methods:Four male patients with total globozoospermia were collected during November 2019 to May 2022 from Reproductive Medical and Genetic Center, the People's Hospital of Guangxi Zhuang Autonomous Region. Peripheral blood samples were collected for genetic detection and the whole exome sequencing to explore the pathogenic genes. Semen characteristics, sperm morphology and ultrastructure were analyzed. Four patients were treated with intracytoplasmic sperm injection (ICSI) combined with artificial oocyte activation (AOA). Conventional ICSI cycles ( n=9) were selected as control group, and the development dynamic parameters were monitored by Time-lapse. The fertilization and embryo development parameters, developmental dynamic parameters and clinical outcomes were analyzed between the two groups. Results:Four patients were complicated with low sperm motility and increased sperm DNA fragmentation. Sperm morphology analysis and acrosome fluorescence staining represented that all the spermatozoas were with a small round head lacked of acrosome. By the transmission electron microscope, it was observed that round-headed spermatozoas were lacked of acrosome completely, loose chromatin structure, vacuolation and other abnormal changes in the head, mitochondrial sheath in neck were reduced arranged in disorder, and the structure of "9+2" of the flagellar axial filament was incomplete. Of the 4 patients, 1 was homozygous deletion of DPY19L2 gene and 1 was homozygous frameshift mutation of DPY19L2 gene. There were no significance differences in fertilization rate, two pronuclei fertilization rate, day 3 high-quality embryo rate, day 6 blastocyst formation rate and day 6 high-quality blastocyst formation rate between total globozoospermia group and control group (all P>0.05). The developmental dynamic parameters as the time at which the second polar body is extruded, the time when both (or the last) PN disappear, two to six discrete cells in the total globozoospermia group were significantly earlier than those in control group, and the difference was statistically significant (all P<0.05). There was no significant difference in the embryo cleavage patterns between the two groups ( P>0.05). Among the 4 patients with total globozoospermia, 2 live births with signal healthy male baby were achieved by fresh embryo transfer, and 2 live births with one signal healthy male baby and one healthy female baby were achieved by frozen-thawed embryo transfer respectively. Conclusion:Abnormal morphology characteristics of spermatozoas from patients with total globozoospermia are obvious, patients with total globozoospermia could have favorable clinical outcomes following ICSI combined with AOA.

Objective:To analyze the clinical phenotype, sperm morphological characteristics and assisted reproductive therapy efficiency in patients with total globozoospermia.Methods:Four male patients with total globozoospermia were collected during November 2019 to May 2022 from Reproductive Medical and Genetic Center, the People's Hospital of Guangxi Zhuang Autonomous Region. Peripheral blood samples were collected for genetic detection and the whole exome sequencing to explore the pathogenic genes. Semen characteristics, sperm morphology and ultrastructure were analyzed. Four patients were treated with intracytoplasmic sperm injection (ICSI) combined with artificial oocyte activation (AOA). Conventional ICSI cycles ( n=9) were selected as control group, and the development dynamic parameters were monitored by Time-lapse. The fertilization and embryo development parameters, developmental dynamic parameters and clinical outcomes were analyzed between the two groups. Results:Four patients were complicated with low sperm motility and increased sperm DNA fragmentation. Sperm morphology analysis and acrosome fluorescence staining represented that all the spermatozoas were with a small round head lacked of acrosome. By the transmission electron microscope, it was observed that round-headed spermatozoas were lacked of acrosome completely, loose chromatin structure, vacuolation and other abnormal changes in the head, mitochondrial sheath in neck were reduced arranged in disorder, and the structure of "9+2" of the flagellar axial filament was incomplete. Of the 4 patients, 1 was homozygous deletion of DPY19L2 gene and 1 was homozygous frameshift mutation of DPY19L2 gene. There were no significance differences in fertilization rate, two pronuclei fertilization rate, day 3 high-quality embryo rate, day 6 blastocyst formation rate and day 6 high-quality blastocyst formation rate between total globozoospermia group and control group (all P>0.05). The developmental dynamic parameters as the time at which the second polar body is extruded, the time when both (or the last) PN disappear, two to six discrete cells in the total globozoospermia group were significantly earlier than those in control group, and the difference was statistically significant (all P<0.05). There was no significant difference in the embryo cleavage patterns between the two groups ( P>0.05). Among the 4 patients with total globozoospermia, 2 live births with signal healthy male baby were achieved by fresh embryo transfer, and 2 live births with one signal healthy male baby and one healthy female baby were achieved by frozen-thawed embryo transfer respectively. Conclusion:Abnormal morphology characteristics of spermatozoas from patients with total globozoospermia are obvious, patients with total globozoospermia could have favorable clinical outcomes following ICSI combined with AOA.

Objective:To investigate whether the early developmental dynamic phenotypes and kinetic parameters of embryos used for embryo selection are affected by the differences in the components of culture media utilized.Methods:The clinical data of patients undergoing in vitro fertilization (IVF) at Center for Reproductive Medicine and Genetics of People's Hospital of Guangxi Zhuang Autonomous Region from October 2016 to December 2018 were analyzed in a retrospective cohort study. According to the different culture media utilized, IVF cycles were divided into Cook group and Vitrolife group. After 1∶1 propensity-score matching (PSM), 59 IVF cycles were included in each group. Time-lapse imaging technology was used to analyze the early developmental dynamics of normal fertilized embryos between insemination and 68 h post insemination. Seven developmental dynamic phenotypes of embryos were annotated and the differences in the composition of dynamic phenotypes were compared between the two groups. Thirteen early developmental kinetic parameters were calculated, and the differences in the kinetic parameters of normal dynamic phenotypic embryos between the two groups were compared. According to the two published time-lapse embryo selection algorithms, the hierarchical distribution of normal dynamic phenotypic embryos of two groups was compared. Results:1) In Cook group, the composition of developmental dynamic phenotypes of embryos were 54.0% normal phenotype, 3.0% abnormal first cytokinesis (A1 cyt), 17.4% abnormal cleavage (AC), 5.2% reverse cleavage (RC), 3.2% chaotic cleavage (CC), 3.5% multinucleation (Mn) and 13.7% mixed phenotype, which were 49.3%, 4.0%, 19.1%, 7.5%, 2.1%, 6.4% and 11.6% in Vitrolife group, respectively. No statistically significant differences were observed between the two groups for the composition of dynamic developmental phenotypes ( P>0.05). 2) Compared with Vitrolife group, the 13 developmental kinetic parameters (tPNa, tPNf, t2, t3, t4, t5, t6, t7, t8, cc2, s2, t5_PNf and t8_PNf) of normal dynamic phenotype embryos in Cook group were slightly longer, and the average number of blastomeres in 68 h post insemination (EB68hpi) was less, but the differences were not statistically significant (all P>0.05). 3) No significant difference in hierarchical distribution of embryos was observed between Vitrolife group and Cook group according to algorithm A ( P>0.05). The difference of embryo hierarchical distribution between the two groups was statistically significant according to algorithm B ( P=0.040), the proportion of grade A + embryos in Vitrolife group was higher than that in Cook group [59.8% (125/209) vs. 43.3% (94/217)], and grade C proportion was lower [9.6% (20/209) vs. 20.3% (44/217)]. Conclusion:Although the early developmental dynamic phenotypes and kinetic parameters of embryos were not affected by the differences between Cook and Vitrolife sequential culture media, the applicability of different time-lapse embryo selection algorithms to the culture media is different, the embryo culture media utilized should be considered when selecting or constructing the algorithms.

Objective:To investigate whether the early developmental dynamic phenotypes and kinetic parameters of embryos used for embryo selection are affected by the differences in the components of culture media utilized.Methods:The clinical data of patients undergoing in vitro fertilization (IVF) at Center for Reproductive Medicine and Genetics of People's Hospital of Guangxi Zhuang Autonomous Region from October 2016 to December 2018 were analyzed in a retrospective cohort study. According to the different culture media utilized, IVF cycles were divided into Cook group and Vitrolife group. After 1∶1 propensity-score matching (PSM), 59 IVF cycles were included in each group. Time-lapse imaging technology was used to analyze the early developmental dynamics of normal fertilized embryos between insemination and 68 h post insemination. Seven developmental dynamic phenotypes of embryos were annotated and the differences in the composition of dynamic phenotypes were compared between the two groups. Thirteen early developmental kinetic parameters were calculated, and the differences in the kinetic parameters of normal dynamic phenotypic embryos between the two groups were compared. According to the two published time-lapse embryo selection algorithms, the hierarchical distribution of normal dynamic phenotypic embryos of two groups was compared. Results:1) In Cook group, the composition of developmental dynamic phenotypes of embryos were 54.0% normal phenotype, 3.0% abnormal first cytokinesis (A1 cyt), 17.4% abnormal cleavage (AC), 5.2% reverse cleavage (RC), 3.2% chaotic cleavage (CC), 3.5% multinucleation (Mn) and 13.7% mixed phenotype, which were 49.3%, 4.0%, 19.1%, 7.5%, 2.1%, 6.4% and 11.6% in Vitrolife group, respectively. No statistically significant differences were observed between the two groups for the composition of dynamic developmental phenotypes ( P>0.05). 2) Compared with Vitrolife group, the 13 developmental kinetic parameters (tPNa, tPNf, t2, t3, t4, t5, t6, t7, t8, cc2, s2, t5_PNf and t8_PNf) of normal dynamic phenotype embryos in Cook group were slightly longer, and the average number of blastomeres in 68 h post insemination (EB68hpi) was less, but the differences were not statistically significant (all P>0.05). 3) No significant difference in hierarchical distribution of embryos was observed between Vitrolife group and Cook group according to algorithm A ( P>0.05). The difference of embryo hierarchical distribution between the two groups was statistically significant according to algorithm B ( P=0.040), the proportion of grade A + embryos in Vitrolife group was higher than that in Cook group [59.8% (125/209) vs. 43.3% (94/217)], and grade C proportion was lower [9.6% (20/209) vs. 20.3% (44/217)]. Conclusion:Although the early developmental dynamic phenotypes and kinetic parameters of embryos were not affected by the differences between Cook and Vitrolife sequential culture media, the applicability of different time-lapse embryo selection algorithms to the culture media is different, the embryo culture media utilized should be considered when selecting or constructing the algorithms.

Objective:To establish a technical system for early judgment and rescue artificial oocyte activation of oocytes after fertilization failed, intracytoplasmic sperm injection (ICSI) and to explore the activation efficiency and application value.Methods:Firstly, a retrospective analysis was performed in 150 ICSI cycles cultured by Time-lapse system during January 2017 to March 2018 from Reproductive Medical and Genetic Center, the People's Hospital of Guangxi Zhuang Autonomous Region, the time distribution of the second polar body (Pb2) exclusion and its relationship with fertilization and embryo development outcomes, the feasibility of Pb2 exclusion as an early indicator of fertilization failure in ICSI was discussed. Secondly, 225 fertilization failed oocytes from 93 ICSI cycles during March 2018 to June 2019 were collected for randomized controlled trials according to different artificial activation modes. They were divided into three groups including non-activation group (NAOA group), rescue activation group (RAOA group) and late activation group (LAOA group). At the same time, the rest of injected oocytes from ICSI cycles include in this study were used as control group. Fertilization and embryo development parameters were used to explore the efficiency and application value of rescue artificial activation for after fertilization failure oocytes ICSI.Results:Time-lapse monitoring showed that the fertilization rate and 2PN fertilization rate were 99.91% and 97.76% in the Pb2-exclusion group, 0.03% and 0% in the without Pb2-exclusion group after ICSI, with significant differences between the two groups ( P<0.001); the exclusion time of Pb2 after ICSI was (3.04±1.45) h, and the distribution and proportion of Pb2 exclusion time were 0-3 h (58.00%), 3-5 h (36.29%), 5-8 h (3.92%) and >8 h (1.21%). The Pb2 exclusion rate, the fertilization rate and the 2PN fertilization rate in NAOA group were all 0%, and the 2PN fertilization rate (36.00%), day 3 (D3) high-quality embryo rate (8.00%), day 5 (D5) blastocyst formation rate (0%) and D5 high-quality blastocyst formation rate (0%) in LAOA group were significantly lower than those in RAOA group (60.00%, 44.19%, 51.16%, 25.58%) ( P=0.005, P=0.002, P<0.001, P=0.005), and also in control group (97.63%, 48.62%, 63.23%, 37.94%) ( P<0.001); the Pb2 exclusion rate (84.00%), the fertilization rate (81.33%), the 2PN fertilization rate (60.00%) and the 2PN cleavage rate (95.56%) in RAOA group were significantly lower than those in control group (100.00%, 99.68%, 97.63%, 99.51%) ( P<0.001, P<0.001, P<0.001, P=0.04). However, there was no significant difference in D3 high-quality embryo rate, D5 blastocyst formation rate and D5 high-quality blastocyst formation rate between the two groups ( P>0.05). Conclusion:Time-lapse monitoring of the Pb2 exclusion can be used as an early judgment indicator of fertilization failure in ICSI cycles. Rescue artificial activation of fertilization failed oocytes in 5 h after ICSI can achieve normal fertilization and acceptable embryo development outcomes.

Objective:To establish a technical system for early judgment and rescue artificial oocyte activation of oocytes after fertilization failed, intracytoplasmic sperm injection (ICSI) and to explore the activation efficiency and application value.Methods:Firstly, a retrospective analysis was performed in 150 ICSI cycles cultured by Time-lapse system during January 2017 to March 2018 from Reproductive Medical and Genetic Center, the People's Hospital of Guangxi Zhuang Autonomous Region, the time distribution of the second polar body (Pb2) exclusion and its relationship with fertilization and embryo development outcomes, the feasibility of Pb2 exclusion as an early indicator of fertilization failure in ICSI was discussed. Secondly, 225 fertilization failed oocytes from 93 ICSI cycles during March 2018 to June 2019 were collected for randomized controlled trials according to different artificial activation modes. They were divided into three groups including non-activation group (NAOA group), rescue activation group (RAOA group) and late activation group (LAOA group). At the same time, the rest of injected oocytes from ICSI cycles include in this study were used as control group. Fertilization and embryo development parameters were used to explore the efficiency and application value of rescue artificial activation for after fertilization failure oocytes ICSI.Results:Time-lapse monitoring showed that the fertilization rate and 2PN fertilization rate were 99.91% and 97.76% in the Pb2-exclusion group, 0.03% and 0% in the without Pb2-exclusion group after ICSI, with significant differences between the two groups ( P<0.001); the exclusion time of Pb2 after ICSI was (3.04±1.45) h, and the distribution and proportion of Pb2 exclusion time were 0-3 h (58.00%), 3-5 h (36.29%), 5-8 h (3.92%) and >8 h (1.21%). The Pb2 exclusion rate, the fertilization rate and the 2PN fertilization rate in NAOA group were all 0%, and the 2PN fertilization rate (36.00%), day 3 (D3) high-quality embryo rate (8.00%), day 5 (D5) blastocyst formation rate (0%) and D5 high-quality blastocyst formation rate (0%) in LAOA group were significantly lower than those in RAOA group (60.00%, 44.19%, 51.16%, 25.58%) ( P=0.005, P=0.002, P<0.001, P=0.005), and also in control group (97.63%, 48.62%, 63.23%, 37.94%) ( P<0.001); the Pb2 exclusion rate (84.00%), the fertilization rate (81.33%), the 2PN fertilization rate (60.00%) and the 2PN cleavage rate (95.56%) in RAOA group were significantly lower than those in control group (100.00%, 99.68%, 97.63%, 99.51%) ( P<0.001, P<0.001, P<0.001, P=0.04). However, there was no significant difference in D3 high-quality embryo rate, D5 blastocyst formation rate and D5 high-quality blastocyst formation rate between the two groups ( P>0.05). Conclusion:Time-lapse monitoring of the Pb2 exclusion can be used as an early judgment indicator of fertilization failure in ICSI cycles. Rescue artificial activation of fertilization failed oocytes in 5 h after ICSI can achieve normal fertilization and acceptable embryo development outcomes.

Objective To comparative analysis the intracytoplasmic sperm injection (ICSI)result and rate of sperm DNA in-tegrity (DNA fragmentation index,DFI)about testicular and epididymis sperm.Methods Totally 183 obstructed azoospermia pa-tients were choosed to use ICSI.80 cycles by PESA and 103 cycles by TESA,compared two groups of sperm DNA integrity rate and ICSI outcome.Results Sperm DNA integrity rate,fertilization rate,cleavage rate,good-qualityembryo rate and pregnancy rate com-pared with no difference by ICSI(P >0.05).Conclusion DNA integrity rate and ICSI outcomes of the testis and epididymis sperm have no significant differences,clinicians can be based on personal experiences or patients,wills to select sperm for ICSI.