Objective To investigate the relationship between mtND4 point mutation in sperms and asthenospermia. Methods Fifty-six asthenospermia cases and 44 control cases were collected using the WHO criterion for defining asthenospermia, the regions of mtND4 gene were amplified by using PCR of 3 pairs primers. Consequently, the point mutation, missense mutation and multiple single nucleotide polymorphisms (SNP) were analyzed by employing sequencing technology and bioinformatics tools. Results Six mutations never before identified were found. The frequency of single point mutation T10873C and T11944C in the control group were significantly higher than those in the asthenospermia group (P＜0.05). Eight cases involved T10873C or T11944C among the 10 cases in control groups with missense mutations were found. But, there were only 2 cases with such mutation in the 10 asthenospermia cases with missense mutations (P＜0.05). The previous 20 cases of missense mutations can be described as either multiple SNP group (with T10873C or T11944C) or nonmultiple SNP group. The percentage of a range and a plus b range of multiple SNP group of sperm was significantly higher than the non-multiple SNP group(P＜0.05). Conclusions mtND4 gene mutation, especially the missense mutation may induce loss of sperm motility. The mutations of T10873C and T11944C may be useful for sperm motility or counteract the influence for the sperm motility caused by these harmful mutations.

Objective To investigate the correlation between seminal plasma hepatitis B virus (HBV) DNA copy and semen parameters and sperm DNA fragmentation (SDF).Methods The seminal plasma HBV-DNA was detected by the real-time PCR in 148 infertility males,and those with serum HBV-DNA above (positive) or below (negative) 5.0 × 102U/ml were analyzed respectively by semen parameters,sperm morphology and sperm DNA fragmentation (SDF).Results Of 148 male,60 (40.5％) were seminal plasma HBV-DNA positive,and of 60 positive patients,56 (93.3％) were serum hepatitis B e antigen(HBeAg) positive,which was higher than those of seminal plasma HBV-DNA negative males (31cases,35.2％).Serum HBeAg and HBV-DNA in seminal plasma HBV-DNA positive patients were 845.7(0.2 ～ 1455.0) S/CO and (1.7 ± 1.1) × 108U/ml,which were higher than those of HBV-DNA negative patients [HBeAg:0.1 (0.1 ～ 1374.0) S/CO;HBV-DNA:(2.3 ± 1.1) × 107 U/ml,P ＜ 0.01].Seminal plasma HBV-DNA positive patients exhibited lower semen volume,sperm concentration,the percentage of forward moving sperm and less normal morphology compared to HBV-DNA negative patients [(2.44±1.2)mlvs.(3.07±1.3)ml,(66.8±49.1) ×106/mlvs.(87.1 ±65.4) ×106/ml,(54.3± 16.1)％ vs.(59.1 ±15.3)％,(3.77 ±2.8)％ vs.(6.15 ±4.2)％,P＜0.05].The number of patients with teratozoospermia was significantly higher in seminal plasma HBV-DNA positive patients (56.7％ versus 34.1％,(P ＜ 0.01).The SDF in seminal plasma HBV-DNA positive patients was(18.1 ± 12.3)％,while it was(14.4 ± 8.4)％ in negative patients,and the difference of SDF in these two groups was significantly (t =2.197,P ＜ 0.05).Conclusion Seminal plasma HBV-DNA positive could affect the semen parameters,sperm morphology and SDF.

Objective To investigate the impact of hepatitis B virus (HBV) infection on semen parameters,sperm DNA integrity,acrosin activity and sperm-nucleoprotein transition.Methods Semen samples from 527 subjects including 273 hepatitis B surface antigen (HBsAg) positive and 254 HBsAg negative,who sought medical attention and received in-vitro feritilization in reproductive medicine center of First Hospital of Wenzhou Medical University from Jan 2011 to Oct 2012 were collected.Semen parameters,sperm DNA fragmentation index (DFI),sperm-nucleoprotein transition and acrosin activity of both HBsAg-positive and HBsAg-negative subjects were analyzed.Results Semen parameters of both groups were within the normal range,but sperm concentration and percentage of forward moving sperms of HBsAg positive group were significantly lower than those of HBsAg negative group (P=0.000),while percentage of static sperms of HBsAg positive group were significantly higher than that of HBsAg negative group (P =0.000).DFI in HBsAg positive and negative group were (17.85 ± 0.70) ％ and (11.85 ± 0.50) ％,respectively,which was significantly different (t=6.951,P=0.000).Percentage of sperms with normal morphology in both groups were within the normal range,but sperms with neck and tail deformity in the HBsAg positive group was significantly higer than those in HBsAg negative group (all P＜0.05).Acrosin activity of sperms in HBsAg positive group was significantly lower than that in HBsAg negative group (t=3.756,P=0.000).Linear regression analysis indicated that serum HBsAg level was reversely correlated with sperm concentration (r=-0.140,P =0.021),but positively correlated to DFI (r =0.151,P =0.014).Conclusions HBV infection not only affects the routine semen parameters and sperm morphology,but also compromises sperm function including impaired DFI and acrosin activity.However,the impact of anti-HBV agents on sperm quality and male fertility requires further research.

Objective To investigate the correlation between sperm-nucleporotein transition and sperm parameters and embryo development,also to evaluate the influence of pregnancy out comes of assisted reproductive technology (ART).Methods Sperm-nucleoprotein transition assay of a total of 676 patients underwent ART treatment were detected by aniline blue staining,and the correlation analysis between spermnucleoprotein transition and sperm parameters,DNA damage,acrosin activity,fertilization rate,cleavage rate,quality of early embryo development as well as blastocyst formation rate was performed.Results The sperm concentration,(a+b) ％ sperm,sperm count and acrosin activity was (66.5±4.6) × 109/L,(149.2±9.9)×109/L,(51.2±1.3)％ and (72.2±3.3) mU/106 sperm in abnormal group,and (91.9±2.7) ×109/L,(240.0±8.0) ×109/L,(57.3±0.8)％ and (85.7±1.9) mU/106 sperm in normal group,which reached significant difference (P＜0.01).DNA fragmentation index (DFI) was (17.3± 1.0)％ in abnormal group,which was significantly higher than (14.6±0.5)％ in normal group.The cleavage rate of 95.0％,D3/D5 high quality embryo rates of 34.2％ and 1.28％,D5 blastocyst formation rate and the total rate of blastocyst formation rate of 22.4％ and 38.6％ in abnormal group,which were significantly lower than that in normal group (96.9％,38.2％,2.70％,27.9％ and 46.4％) (P＜0.01).The rate of spontaneous abortion was 12.3％ in abnormal group,which was significantly higher than that in normal group (4.7％) (P＜0.01).However,there was no significant difference in biochemical pregnancy rate and ectopic pregnancy rate between the 2 groups (P＞0.05).Conclusions Sperm-nucleoprotein transition was positively related with sperm parameters,DNA damage,acrosin activity,and also has an adverse effect on embryo development and the outcomes of ART.It is suggested that the sperm-nucleoprotein transition should be detected before ART.