OBJECTIVE: To discuss seminal plasma oxidative stress and sperm DNA oxidative damage in patients with idiopathic asthenozoospermia. METHODS: Infertile couples were selected from the clinic outpatients of the Reproductive Center of Xiangya Hospital, Central-South University from December 2010 to March 2011. Fresh semen of 28 men with idiopathic asthenozoospermia was collected as an experiment group, and 24 fertile men with normal semen and normal reproductive history served as a control group. Level of reactive oxygen species (ROS) in the seminal plasma was assessed with luminer chemiluminescence method. Density of sperm DNA oxidation product 8-hydroxy-2'-deoxyguanosine (8-OHdG) was assessed with enzyme linked immunosorbent assay. RESULTS: 1) ROS level in the experiment group was higher than that in the control group (P<0.01). There was negative correlation between the percentage of progressive motility spermatozoa and the ROS level in the seminal plasma in the 2 groups (r=-0.72, P<0.01). 2) Density of sperm 8-OHdG in the experiment group was higher than that in the control group (P<0.01). There was negative correlation between the percentage of progressive motility spermatozoa and the density of sperm 8-OHdG (r=-0.73, P<0.01). 3) There was positive correlation between the ROS level in the seminal plasma and the density of sperm 8-OHdG (r=0.77, P<0.01). CONCLUSION There is sperm DNA oxidative damage in patients with idiopathic asthenozoospermia, which may be related with the oxidative stress. Excessive generation of reactive oxygen species may be a cause of low sperm motility in patients with idiopathic asthenozoospermia.
8-Hydroxy-2'-Deoxyguanosine ; Adult ; Asthenozoospermia ; etiology ; genetics ; metabolism ; Case-Control Studies ; DNA Damage ; Deoxyguanosine ; analogs & derivatives ; metabolism ; Humans ; Infertility, Male ; genetics ; Male ; Oxidative Stress ; physiology ; Spermatozoa ; metabolism ; Young Adult

Objective:To investigate the expression of H19 long non-coding RNA (IncRNA) and zinc finger E-box-binding protein i (ZEB1) in the trophoblast of women with spontaneous abortion.Methods:A total of 20 women underwent miscarriage were enrolled as a miscarriage group,while 20 women underwent artificial abortion were enrolled as a control group.Reverse transcriptional fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of iH19 lncRNA and miR-200,and Western blot was used to detect the expression of ZEB1.Results:Compared to the control group,a tendency of decreased H19 was detected in the miscarriage group,with no significant difference (1.28±0.08 vs 1.66±0.21,P=0.091).There was no significant difference in miR-200 between the 2 groups (0.75±0.35 vs 0.58±0.33,P=0.0.533).The expression of ZEB 1 was significantly decreased in the miscarriage group compared to that in the control group (0.22±0.03 vs 0.41±0.04,P=0.0003).The expression of i19 lncRNA and ZEB1 was positively correlated (r=0.529,P=0.0005),and the correlation between H19 lncRNA and miR-200 had no statistical significance (r=0.0293,P=0.858).The correlation between miR-200 and ZEB1 also had no statistical significance (r=-0.132,P=0.416).Conclusion:Down-regulation of ZEB1 in the trophoblast might be related to miscarriage.H19 lncRNA may regulate the expression of ZEB1.

OBJECTIVE: To explore the relationship between peroxiredoxin I expression and seminal reactive oxygen species (ROS) in patients with idiopathic asthenozoospermia. METHODS: Twenty-six infertile male patients were selected from the Reproductive Medical Center, Xiangya Hospital, from September to December in 2012. Fresh semen was collected from an experimental group (26 idiopathic asthenozoospermia patients) and a control group (15 men with fertility history and normal semen). Luminol chemiluminescence method was applied to detect the seminal ROS level. Western blot was used to detect the peroxiredoxin I expression. RESULTS: 1)The seminal ROS level in the experimental group was significantly increased compared with that in the control group (P<0.05), and the seminal ROS level was negatively correlated with mobility of the sperm (r=-0.777, P<0.01). 2) Compared with the control group, the peroxiredoxin I expression was significantly downregulated in the experimental group (P<0.05). The content of sperm peroxiredoxin I in the 2 groups was negatively correlated with the seminal ROS level (r=-0.474, P<0.01). 3) The content of peroxiredoxin I had a positive correlation with human sperm motility(r=0.779, P<0.01). CONCLUSION The decline of peroxiredoxin I expression may be one of the crucial factors that leads to idiopathic asthenozoospermia. High level of ROS may be one of the main reasons for sperm vitality decline in patients with idiopathic asthenozoospermia.
Asthenozoospermia ; Case-Control Studies ; Humans ; Male ; Peroxiredoxins ; metabolism ; Reactive Oxygen Species ; chemistry ; Semen ; chemistry ; Sperm Motility ; Spermatozoa ; metabolism

OBJECTIVE: To explore the different effect of short 7-day gonadotropin releasing hormone agonist (GnRHa) protocol and GnRHa long protocol on the insulin-like growth factor II(IGF-II) and insulin-like growth factor binding protein-4 (IGFBP-4) levels in follicular fluid. METHODS: Eighty-eight infertile patients due to tubal factors were included in this study. They were randomly divided into a short 7-day GnRHa protocol group and a GnRHa long protocol group (n = 44). Follicular fluid was obtained from dominant follicles during oocyte retrieval. Levels of IGF-II and IGFBP-4 in the follicular fluid were detected by radioimmunoassay and enzyme-linked immunosorbent assay respectively. RESULTS: Duration of controlled ovarian stimulation was significantly shorter and the injected dosages of gonadotropin were significantly lower in the short 7-day protocol group. The differences in serum levels of estradiol and estradiol per mature follicle on the day of human chorionic gonadotropin injection between the two groups were not significant. The concentrations of IGF-II and IGFBP-4 in the follicular fluid of the short 7-day protocol group were significantly lower,while the difference of the ratio of IGF-II/IGFBP-4 between the two groups was not significant. Linear correlation analysis showed that IGF-II level in the follicular fluid was positively correlated to the total dose of gonadotropin. CONCLUSION The short 7-day and long GnRHa protocols may affect the concentrations of IGF-II and IGFBP-4 in the follicular fluid. However, changes of IGF-II and IGFBP-4 concentrations do not contribute to different clinical outcomes.
Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Follicular Fluid ; chemistry ; Gonadotropin-Releasing Hormone ; administration & dosage ; agonists ; Humans ; Insulin-Like Growth Factor Binding Protein 4 ; analysis ; Insulin-Like Growth Factor II ; analysis ; Ovulation Induction ; methods

OBJECTIVE: To evaluate the impact of sperm source and sperm parameters on the outcome of intracytoplasmic sperm injection (ICSI). METHODS: This retrospective study included 433 ICSI cycles from June 2005 to December 2008 in Reproductive Medical Center of Xiangya Hospital. The patients were divided into 2 major groups according to the source of spermatozoa used for ICSI: ejaculated (group A, n=336) and epididymal (group B, n=97). Group A was divided into 3 subgroups according to the sperm parameters: normal (Group A1, n=95), single parameter defect (Group A2, n=119), and multiple parameter defect (Group A3, n=122). RESULTS: The basic characteristics among the 4 groups had no statistic difference (P>0.05), and the difference in the fertilization rate, normal fertilization rate, cleaving embryo rate,good quality embryo rate, implanted rate, clinical pregnancy rate, and early abortion rate among the 4 groups were not significant (P>0.05). CONCLUSION The outcome is similar no matter whether the spermatozoa is from ejaculated sperm or epididymis. ICSI can treat male infertility of various factors, and the outcome is the same with one or multiple sperm parameter abnormality. ICSI with epididymal spermatozoa through percutaneous epididymal sperm aspiration is effective for infertility due to obstructive azoospermia.
Adult ; Epididymis ; cytology ; physiopathology ; Female ; Fertilization ; Humans ; Infertility, Male ; therapy ; Male ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Sperm Motility ; physiology ; Sperm Retrieval ; Spermatozoa ; physiology

To compare the cumulative live birth rates (CLBR) and the incidence of ovarian hyperstimulation syndrome (OHSS) between fresh embryo transfer (ET) and frozen ET (the freeze-all policy), when oocyte numbers are more than 15 in the first treatment of in vitro fertilization or intracytoplasmic sperm injection, and to evaluate the benefits of the freeze-all policy.
 Methods: We retrospectively analyzed clinical data of 2 842 patients whose oocytes numbers were more than 15, including 1 095 frozen ET patients and 1 747 fresh ET patients. The patients general data, a baseline features, CLBR, and the incidence of OHSS were compared between the 2 groups.
 Results: There were 598 patients in the 2 groups after they experienced the propensity score matching. No significant differences were found in age, infertility causes, body mass index, basal follicle stimulating hormone level, the total days and total dose of using gonadotrophin (Gn) between the 2 groups (all P>0.05). The CLBR of the freeze-all cycles increased along with the number of oocytes (P>0.05), and the oocyte numbers were greater in freeze-all group than those of the fresh ET group (P<0.001). There was no significant difference in CLBR after one complete cycle between the 2 groups (P>0.05), but after the first embryo transfer cycle, the CLBR in freeze-all group was higher than that in the fresh ET cycle group (P<0.05). The incidence of OHSS in patients with freeze-all was significantly lower than that in the patiants with fresh ET (P<0.05).
 Conclusion: Patients with oocytes over 15 and OHSS tendency who accepted the freeze-all strategy can help them to prevent OHSS and they have a higher CLBR than fresh ET cycles.
Birth Rate ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Oocytes ; Pregnancy ; Pregnancy Rate ; Retrospective Studies

OBJECTIVE: To determine the sperm-zona pellucida (ZP) binding and ZP-induced acrosome reaction in patients with unexplained infertility, and to discuss the relationship between ZP-induced acrosome reaction and fertilization rate. METHODS: We compared the fertilization rate and good embryo rate in patients with unexplained infertility after fertilization in 2 ways. Based on the causes of infertility, patients were divided into an unexplained infertility group (Group A) and a pure female tubal factor group (Group B). Oocytes which were obtained by super ovulation from 25 patients with unexplained infertility were randomly divided into 2 groups with conventional in vitro fertilization (IVF) (Group A1) and intracytoplasmic sperm injection (ICSI) fertilization (Group A2). The pure female tubal factor group (Group B) had conventional IVF. We conducted sperm-ZP binding and ZP-induced acrosome reaction experiments with 2 groups of men's sperms separately. We compared the number of sperm-egg binding and ZP-induced acrosome reaction rate and discussed the relationship between the ZP-induced acrosome reaction and fertilization rate, and also the fertilization rate, good embryo rate and pregnancy rate in patients with unexplained infertility after fertilization in 2 ways. RESULTS: The average number of sperm-egg binding (78.29 ± 16.31) and the ZP-induced acrosome reaction rate (55.87 ± 27.69) % in Group A were lower than those of Group B [94.63 ± 6.72, (82.53 ± 17.99)%]. The difference between the average number of sperm-egg binding and the ZP-induced acrosome reaction was significant (P <0.01). The fertilization rate of Group A1 was significantly lower than that of Group B and Group A2 (P <0.01). But there was no significant difference in the good embryo rate among the 3 groups. There was no significant difference between Group A2 and B in fertilization rate and good embryo rate (P <0.05). There was no significant difference in pregnancy rate between Group A and B (P <0.05). Fertilization rate and the rate of acrosome reaction had marked positive correlation with statistical significance (r =0.932, P <0.01). CONCLUSION ZP binding and ZP-induced acrosome reaction are very important experiments in sperm function test for patients with unexplained infertility. It can not only effectively avoid no embryo transferring due to complete failure of fertilization but also get a desirable outcome of pregnancy using half-ICSI fertilization in patients with unexplained infertility.
Acrosome Reaction ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility ; etiology ; therapy ; Male ; Oocytes ; physiology ; Ovulation Induction ; Sperm Injections, Intracytoplasmic ; Sperm-Ovum Interactions ; physiology ; Treatment Outcome ; Zona Pellucida ; physiology

OBJECTIVE: To explore the correlation between sperm-originated miRNAs and embryo quality by detecting the expression levels of miR-21, miR-34c, miR-140 and miR-375 in the sperm from in vitro fertilization (IVF) patients.
 METHODS: The fresh semen specimens were collected from 44 male patients who received the IVF cycle in the Xiangya Hospital Reproductive Center from September to December in 2012. The expression levels of miR-34c, miR-140, miR-21 and miR-375 were detected by real-time fluorescent quantitative PCR. The embryos on day 2 and day 3 after fertilization were divided into the experimental and the control group, with an average embryo scores less or greater than 8, respectively. Then we compared the general and experimental data between the 2 groups respectively and analyzed the correlation between the miRNAs levels in sperm and embryo quality.
 RESULTS: The expression of miR-34c, miR-140, miR-21 and miR-375 in sperms from the experimental group was significantly lower than that in the control group (P<0.05). There was no significant difference in retrieved follicles, metaphase II stage ovocytes, fertilized oocytes, cleavage number and fertilization rate between the experimental group and the control group (P>0.05), while the cleavage rate on day 2 in the experimental group was significantly lower than that of the control group (P<0.05). There was a negative correlation between the expression levels of miRNAs (miR-21, miR-34c, miR-140 and miR-375) and the ratio of fragment on day 2 or day 3. The expression levels of miR-21, miR-34c, miR-140 and miR-375 was positively correlated with the embryo score and the blastomere quantity on day 3, respectively. 
 CONCLUSION The up-regulated levels of miR-21, miR-34c, miR-140 and miR-375 in sperm may function as positive regulators in the development of cleavage stage in embryo and thus influence embryonic quantity.
Fertilization ; Fertilization in Vitro ; Humans ; Male ; MicroRNAs ; metabolism ; Oocytes ; Spermatozoa ; metabolism ; Up-Regulation

OBJECTIVE: To investigate the risk factors associated with early miscarriage among intrauterine singleton pregnancies after treatment with in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI).
 METHODS: A retrospective case-control study was performed on all singleton pregnancies underwent IVF/ICSI from January, 2013 to May, 2014, in Xiangya Hospital. Ninety-six early miscarriage patients served as a case group and 593 pregnancies with live birth served as a control group. We analyzed factors for early miscarriage after IVF/ICSI in two groups.
 RESULTS: Multivariate logistic regression analysis demonstrated that the women age, miscarriage history, and sperm DAN fragmentation index (DFI) were the risk factors for early miscarriage (P<0.05).
 CONCLUSION Miscarriage after treatment with IVF/ICSI is affected by multiple factors. Women at elder age (>30 years old), women with a history of miscarriage or men with higher sperm DFI (≥15%) are the risk.
Abortion, Spontaneous ; Case-Control Studies ; Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Retrospective Studies ; Risk Factors ; Sperm Injections, Intracytoplasmic ; Spermatozoa