The effect of sperm DNA fragmentation on in vitro fertilization outcomes of unexplained infertility

Abstract Background Infertility is caused by heterogeneous risks, but most of them are unexplained. The sperm DNA Fragmentation Index (DFI) was increasingly acknowledged as a parameter for the evaluation of male infertility. This study aimed to investigate the association between sperm DFI and laboratory and clinical outcomes in a population with unexplained infertility. Methods The clinical data of an infertile population was collected for the selection of reproductive patients with unexplained infertility. The authors classified the patients with normal sperm parameters in a control group (DFI < 25%) and an observation group (DFI ≥ 25%) and compared the difference in basal characteristics, laboratory, and clinical outcomes between the two groups. The authors conducted a correlation analysis to examine the relationship between DFI and the number of D3 good-quality embryos, as well as the clinical pregnancy rate and live birth rate. A total of 176 cases were enrolled in the retrospective study. Results The observation group (n = 88) showed advanced male age, lower sperm concentration, progressive motility, and morphology assessment than the control group. In addition, lower No. of D3 good-quality embryos, clinical pregnancy rate, and the live birth rate were shown in the observation group. A negative correlation between the DFI and No. of D3 good-quality embryos (rs = -0.347, p < 0.001) or live birth rate (rs = -0.185, p = 0.028) was shown. Conclusions Sperm DFI was a good indicator for the prediction of D3 good-quality embryos in unexplained infertility couples, but it did not provide sufficient information regarding clinical pregnancy outcome but live pregnancy outcome.

The effect of age and abstinence time on semen quality: a retrospective study / 亚洲男科学杂志(英文版)

This study analyzed the effects of male age and abstinence time on semen quality and explored the best abstinence time for Chinese males among different age groups. Semen parameters, including sperm kinetics, morphology, and DNA fragmentation index (DFI), were reviewed from 2952 men. Samples were divided into six age groups (≤25 years, 26-30 years, 31-35 years, 36-40 years, 41-45 years, and >45 years) and were divided into six groups according to different abstinence time (2 days, 3 days, 4 days, 5 days, 6 days, and 7 days). The differences in semen quality between the groups were compared, and the effect of age and abstinence time on semen quality was analyzed. Significant differences were observed in semen volume, progressive motility (PR), and DFI among the age groups (all P < 0.05), and no significant differences were observed in sperm morphological parameters (all P > 0.05). There were significant differences in semen volume, PR, and DFI among different abstinence time groups (all P < 0.05) and no significant differences in sperm morphological parameters (all P > 0.05). Pearson analysis showed that male age and abstinence time were both significantly correlated with sperm kinetics and DFI (both P < 0.05), while no significant correlation was found with sperm morphological parameters (all P > 0.05). The box plots and histograms of men's age, abstinence time, and semen quality show that most semen quality parameters differ significantly between the 2 days and 7 days abstinence groups and other groups at different ages. Except for the sperm morphology parameters, sperm kinetic parameters and sperm DFI are linearly related to male age and abstinence time.

Random sperm DNA fragmentation index is not associated with clinical outcomes in day-3 frozen embryo transfer / 亚洲男科学杂志(英文版)

Damage to sperm DNA was proposed to play an important role in embryonic development. Previous studies focused on outcomes after fresh embryo transfer, whereas this study investigated the influence of sperm DNA fragmentation index (DFI) on laboratory and clinical outcomes after frozen embryo transfer (FET). This retrospective study examined 381 couples using cleavage-stage FET. Sperm used for intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF) underwent density gradient centrifugation and swim up processing. Sperm DFI had a negative correlation with sperm motility (r = -0.640, P < 0.01), sperm concentration (r = -0.289, P < 0.01), and fertilization rate of IVF cycles (r = -0.247, P < 0.01). Sperm DFI examined before and after density gradient centrifugation/swim up processing was markedly decreased after processing (17.1% vs 2.4%, P < 0.01; 65 randomly picked couples). Sperm progressive motility was significantly reduced in high DFI group compared with low DFI group for both IVF and ICSI (IVF: 46.9% ± 12.4% vs 38.5% ± 12.6%, respectively; ICSI: 37.6% ± 14.1% vs 22.3% ± 17.8%, respectively; both P < 0.01). The fertilization rate was significantly lower in high ( ≥25%) DFI group compared with low (<25%) DFI group using IVF (73.3% ± 23.9% vs 53.2% ± 33.6%, respectively; P < 0.01) but was equivalent in high and low DFI groups using ICSI. Embryonic development and clinical outcomes after FET were equivalent for low and high DFI groups using ICSI or IVF. In this study, sperm DFI did not provide sufficient information regarding embryo development or clinical outcomes for infertile couples using FET.

Influence of the sperm DNA fragmentation index on the outcome of rescue ICSI and the clinical value of rescue ICSI / 中南大学学报(医学版)

OBJECTIVES@#As a remedy for the failure of in vitro fertilization (IVF), rescue intracytoplasmic sperm injection (R-ICSI) has been widely carried out, but it has failed to significantly improve the fertilization rate and clinical pregnancy rate. Sperm DNA fragmentation index (DFI) was highly correlated with pregnancy outcome of artificial assisted reproduction. This study aims to investigate the effect of the sperm DFI on the outcome of R-ICSI and the clinical value of R-ICSI.@*METHODS@#This retrospective analysis was conducted among 140 infertile couples receiving R-ICSI in from January 2014 to December 2019. The subjects were assigned into a total fertilization failure (TFF)+low DFI group (R-ICSI after TFF and DFI<30%) (n=63), a TFF+high DFI group (R-ICSI after TFF and DFI≥30%) (n=16), a partial fertilization failure (PFF)+low DFI group (R-ICSI after PFF and DFI<30%) (n=52), a PFF+high DFI group (R-ICSI after PFF and DFI≥30%) (n=9). All transferred embryos were come from R-ICSI. The general clinical data [infertility duration, male age, female age, basal serum level of follicle stimulating hormone (FSH), basal serum level of luteinizing hormone (LH), antral follicle count, endometrial thickness of human chorionic gonadotropin (HCG) day, and eggs] and R-ICSI cycle outcomes (fertilization rate, normal fertilization rate, cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate and live birth rate) were analyzed. In addition, the effect of R-ICSI on the fertilization outcome of conventional IVF total fertilization failure and partial fertilization failure was explored.@*RESULTS@#There was no significant difference in the general clinical data and R-ICSI cycle outcome between the TFF+low DFI group and the TFF+high DFI group (all P>0.05). There was no significant difference in the general clinical data between the PFF+low DFI group and the PFF+high DFI group (all P>0.05). The fertilization rate and normal fertilization rate in the PFF+low DFI group were significantly higher than those in the PFF+high DFI group (85.40% vs 72.41%, 71.90% vs 58.62%, respectively; both P<0.05). However, there was no significant difference in cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate, and live birth rate between the 2 groups (all P>0.05). The R-ICSI cycle of TFF: A total of 79 fresh cycles, 57 fresh transplant cycles, a total of 761 unfertilized oocytes, and 584 M II oocytes were treated with R-ICSI, the fertilization rate was 83.22%, the normal fertilization rate was 75.51%, the cleavage rate was 98.15%, the good embryo rate was 40.74%, the implantation rate was 30.56%, and the clinical pregnancy rate was 43.86%; 29 live births were obtained. The R-ICSI cycle of PFF: A total of 61 fresh cycles, 31 fresh transplant cycles, a total of 721 unfertilized oocytes, and 546 M II oocytes were treated with R-ICSI; the fertilization rate was 83.33%, the normal fertilization rate was 69.78%, the cleavage rate was 97.36%, the good embryo rate was 44.39%, the implantation rate was 25.42%, and the clinical pregnancy rate was 45.16%; 12 live births were obtained.@*CONCLUSIONS@#In the case of partial fertilization failure of IVF, the sperm DFI affects the fertilization rate and normal fertilization rate of R-ICSI; whether it is a TFF of IVF or PFF of IVF, ICSI can be used as an effective remedy way.

Effects of sperm DNA fragmentation index on semen parameters and pregnancy outcomes in intrauterine insemination / 中华男科学杂志

Objective@#To analyze the correlation of the sperm DNA fragmentation index (DFI) level with semen parameters and pregnancy outcomes of artificial insemination of the husband (AIH) in the cycle of intrauterine insemination (IUI).@*METHODS@#We collected the clinical data on 777 cases of IUI, including female clinical indicators, male semen parameters, sperm DFI and pregnancy outcomes. According to the DFI level, we divided the patients into three groups: DFI < 15%, 15% ≤ DFI < 30% and DFI ≥ 30%.@*RESULTS@#The sperm DFI level was significantly elevated with the increased age of the males (P = 0.002) and closely related to the total number of motile sperm (P = 0.002) and total sperm motility (P = 0.000) before treatment, as well as to sperm concentration (P = 0.000), total sperm motility (P = 0.001) and total number of progressively motile sperm (P = 0.000) after density gradient centrifugation. The rate of clinical pregnancy was decreased in the DFI ≥ 30% group. There were no statistically significant differences between sperm DFI and the rates of clinical pregnancy and abortion.@*CONCLUSIONS@#Male age significantly affects the sperm DFI level. Sperm DFI is closely related to sperm motility and total number of progressively motile sperm, but not to the rates of clinical pregnancy and abortion in patients undergoing IUI. IUI can be used as an effective method of assisted reproduction for male infertility./.

Sperm DNA fragmentation in Chinese couples with unexplained recurrent pregnancy loss / 亚洲男科学杂志(英文版)

We aimed to study the association between sperm DNA fragmentation and recurrent pregnancy loss (RPL) in the Chinese population via a retrospective observational study of Chinese couples who had experienced RPL between May 2013 and August 2018. The study population included 461 men from couples with RPL and 411 men from a control group (couples with clinical pregnancy via in vitro fertilization owing to female causes). Routine semen analysis, sperm chromatin analysis, and microscopic (high-power) morphological analysis were performed using semen samples. Semen samples were assessed for volume, sperm count, and motility. The sperm DNA fragmentation index (DFI) was calculated, and the median DFI was obtained. Men were categorized as having normal (37.8%; DFI ≤ 15.0%), moderate (33.6%; 15.0% < DFI < 30.0%), or severe (28.6%; DFI ≥ 30.0%) DNA fragmentation levels. The percentage of men with severe DNA fragmentation was significantly higher in the RPL (42.3%) group than that in the control group (13.1%), whereas the percentage of men with normal levels of DNA fragmentation was significantly lower in the RPL group (22.8%) than that in the control group (54.7%). Subsequent analysis also demonstrated that the sperm DNA fragmentation rate had a moderate reverse correlation with the sperm progressive motility rate (r = -0.47, P < 0.001) and the total motile sperm count (r = -0.31, P < 0.001). We found a positive correlation between RPL and sperm DNA fragmentation. The results suggest that increased sperm DNA damage is associated with RPL.

Correlation of sperm DNA fragmentation index with semen parameters / 中华男科学杂志

Objective@#To explore the relationship of sperm DNA fragmenation index (DFI) with semen parameters and assess its application value in the evaluation of semen quality.@*METHODS@#A total of 9 694 semen samples were collected and examined for sperm DFI and high DNA stainability (HDS) by flow cytometry-assisted sperm chromatin structure analysis (SCSA). According to the WHO Laboratory Manual for the Examination and Processing of Human Semen (5th Ed), the samples were divided into a normal group and abnormal groups A (sperm concentration ［SC］: ［11.3－14.0］ ×10⁶/ml, total sperm motility ［TSM］: 30%－39%, progressively motile sperm ［PMS］: 24%－31%), B (SC: ［7.5－11.2］ ×10⁶/ml, TSM: 20%－29%, PMS: 16%－23%), C (SC: ［3.8－7.4］ ×10⁶/ml, TSM: 10%－,19% PMS: 8%－15%) and D (SC: ［0－3.7］×10⁶/ml, TSM: 0－9%, PMS: 0－7%), and also into three sperm DFI groups (DFI 30%). The correlation between sperm DFI and seminal parameters was analyzed by Pearson correlation and multiple linear regression analyses.@*RESULTS@#DFI was dramatically lower in the normal than in the abnormal groups (P < 0.01), and increased in proportion to the decrease of semen parameters in the abnormal groups A, B, C and D (P < 0.01). Pearson correlation analysis showed that DFI was correlated positively with age (r = 0.15, P < 0.01), abstinence time (r = 0.10, P < 0.01), semen volume (r = 0.05, P < 0.01) and HDS (r = 0.15, P < 0.01), but negatively with semen pH (r = －0.06, P < 0.01), SC (r = －0.27, P < 0.01), TSM (r = －0.53, P < 0.01), PMS (r = －0.52, P < 0.01) and morphologically normal sperm (r = －0.16, P < 0.01). Multiple linear regression analysis revealed that TSM, SC, age, abstinence time and semen pH were five important variables associated with DFI, with standardized regression coefficients of －0.47, －0.19, 0.12, 0.07, and －0.04, respectively (all P < 0.01).@*CONCLUSIONS@#There is a moderate correlation between sperm DFI and semen parameters, which can be used synergistically for the assessment of semen quality.

Effect on sperm quality of asthenospermia and oligospermia treated with grain-moxibustion combined with medicine therapy / 中国针灸

OBJECTIVE@#To observe the clinical effect of grain-moxibustion combined with medicine therapy for asthenospermia and oligospermia.@*METHODS@#A tatal of 60 patients were randomized into an observation group (30 cases) and a control group (30 cases) according to 1︰1 ratio. In the control group, vitamin E capsules were taken orally one capsule each time, twice a day, and pills 6 g each time, three times a day for a total of 3 months. In the observation group, grain-moxibustion was applied at Guanyuan (CV 4)，Shenshu (BL 23) and Zusanli (ST 36) based on the control group, once a week for 3 months, with a total of 12 times. The sperm concentration and sperm progressive motility were measured by automatic sperm quality analysis system in the two groups, and the clinical effects were compared. Sperm DNA fragmentation index (DFI) in the observation group was measured by sperm nucleus chromosome structure assay (SCSA).@*RESULTS@#①The sperm concentrations and sperm progressive motilities after 1-month, 2-month and 3-month of treatment were increased compared with those before treatment in the two groups (<0.01), and they were increased with time. In the two groups, 2-month and 1-month of treatment, 3-month and 2-month of treatment were compared, the sperm concentrations and sperm progressive motilities were significantly increased (<0.01). The sperm concentrations after 1-month, 2-month and 3-month of treatment in the observation group were higher than those in the control group (<0.01), the sperm progressive motility after 3-month of treatment in the observation group was higher than that in the control group (<0.05). ②After 3-month of treatment，the DFI in the observation group was significantly reduced compared with that before treatment (<0.01). ③The total effective rate in the observation group after 3-month of treatment was 86.7% (26/30), which was superior to 63.3% (19/30) in the control group (<0.05).@*CONCLUSION@#Grain-moxibustion combined with medicine therapy can improve sperm concentration and sperm progressive motility, enhance the integrity of sperm DNA.

Diagnostic Value of DNA Fragmentation Index and Acrosin Activity in Male Infertility / 中山大学学报(医学科学版)

[Objective]We aimed to explore the diagnostic value of DNA fragmentation index(DFI)and acrosin activi-ty in male infertility.[Methods]Nine hundred and two semen samples were collected from patients and assessed for the DNA fragmentation index by sperm chromatin dispersion test,acrosin activity,as well as standard sperm parameters according to the WHO criteria. Statistical analysis was performed with SPSS.[Results]Statistically significant differences were observed in age,sperm concentration,total motility,progressive rate and acrosin activity among different group of sperm DNA dam-age(≤10%,11~20%,21~30%,≥31%).Sperm acrosin activity also showed difference in sperm concentration,total num-ber,total motility,progressive rate,normal morphology rate,teratozoospermia index(TZI)and sperm DFI.The DNA frag-mentation rate and sperm concentration,sperm motility,forward motility rate,total sperm count and acrosin activity was neg-atively correlated,while it is negative correlatd with TZI. Acrosin activity and sperm concentration,sperm motility,forward motility rate,sperm count and normal morphology rate was positively correlated,while it is negative correlated with the ab-normal rate of head,sperm deformity index,DNA fragmentation rate.[Conclusion]Sperm DNA damage and acrosin activity could partly reflect the quality of sperm. Moreover,sperm DFI maypredict the sperm motility part while the acrosin activity more likely related to sperm morphology.

Combined therapy of Compound Xuanju Capsules and traditional Chinese medicinal formula for infertility in male smokers / 中华男科学杂志

<p><b>Objective</b>To investigate the clinical effects of the combined therapy of Compound Xuanju Capsules (CXJC) and traditional Chinese medicinal formula on infertility in male smokers.</p><p><b>METHODS</b>A total of 176 male infertility patients were divided into a smoking and a non-smoking group and the former further divided into mild, moderate and heavy smokers according to the daily consumption of cigarettes and the length of smoking history. The patients were treated with CXJC combined with traditional Chinese medicinal formula for 3 four-week courses and the therapeutic results were evaluated by comparing the indicators of the traditional Chinese medicine (TCM) syndrome, routine semen parameters, sperm morphology, and sperm DNA fragmentation index (DFI) among different groups before and after treatment.</p><p><b>RESULTS</b>The baseline TCM syndrome scores were remarkably higher in the heavy smokers than in the non-smoking group (P < 0.05) but showed no statistically significant differences between the mild and moderate smokers (P > 0.05). The baseline percentage of sperm head defects and DFI were also markedly higher in the heavy and moderate smokers than in the non-smoking group (P < 0.05). Compared with the baseline, significant improvement was achieved after treatment in the TCM syndrome, routine semen parameters, sperm morphology and sperm DFI, especially in the heavy smokers in the percentages of grade a+b sperm (［17.12 ± 2.54］ vs ［30.15 ± 3.10］%, P < 0.05), morphologically normal sperm (［15.54 ± 1.98］ vs ［26.82 ± 3.52］%, P < 0.05), and head-defective sperm (［27.02 ± 2.14］ vs ［22.07 ± 1.52］%, P < 0.05).</p><p><b>CONCLUSIONS</b>Sperm quality is significantly decreased while the risk of infertility remarkably increased in moderate and heavy smokers. The combined therapy of CXJC and traditional Chinese medicinal formula can effectively improve semen quality, sperm morphology and sperm DFI in male smokers with infertility, though more evidence is to be collected from further studies.</p>

Correlation of sperm DNA fragmentation index with age and semen parameters in infertile men / 中华男科学杂志

<p><b>Objective</b>To explore the correlation of the sperm DNA fragmentation index (DFI) with age, sperm concentration and sperm motility in infertile men.</p><p><b>METHODS</b>We collected semen samples from 531 infertile males in our hospital from January 2016 to June 2017. We determined the semen parameters using the computer-assisted semen analysis system, measured the sperm DFI by sperm chromatin structure assay, and analyzed the correlation of the sperm DFI with the age, sperm concentration and sperm motility of the patients.</p><p><b>RESULTS</b>With the increase of age, the infertile males showed a significantly decreased proportion of the sperm with a DFI ≤15% and elevated proportion of the sperm with a DFI ≥25%, with a positive correlation between age and sperm DFI (r = 0.653, P < 0.01). With the increase of sperm concentration and motility, however, the proportion of the sperm with a DFI ≤15% was remarkably increased while that of the sperm with 15%<DFI<25% and ≥25% markedly reduced, both sperm concentration and motility negatively correlated with the sperm DFI (r = －0.246, P < 0.01 and r = －0.406, P < 0.01).</p><p><b>CONCLUSIONS</b>The sperm DFI is significantly correlated with age, sperm concentration and sperm motility, and therefore can be used as an important index for the evaluation of semen quality. A comprehensive analysis of the sperm DFI and semen parameters may contribute to an accurate assessment of male fertility.</p>

Effects of sperm morphology, sperm DNA fragment index and seminal plasma zinc on pregnancy outcome of IVF-ET / 中国生化药物杂志

Objective To investigate the influence of sperm morphology, sperm DNA fragmentation index and seminal plasma zinc on pregnancy outcome of in vitro fertilization and embryo transfer (IVF-ET).MethodsA total of 341 infertile couples underwent IVF-ET were selected from January 2016 to June 2016 in our hospital.Sperm morphology, sperm DNA fragmentation index and seminal plasma zinc level were compared according to pregnancy.ResultsIn 341 cases, 204 cases pregnancy and 137 cases of no pregnancy, with the pregnancy rate of 59.8%(204/341).Compared with the pregnancy group, the percentage of normal sperm percentage was low, the abnormal sperm index was higher in the non-pregnant group (P<0.05), the sperm DNA index was higher (P<0.05).The percentage of normal sperm, sperm DNA fragmentation index and seminal plasma zinc and pregnancy rate in sperm morphology were linear (P<0.05), And there was a negative correlation between abnormal sperm index and sperm DNA fragment index (P<0.05), and other indicators were positively correlated (P<0.05).ConclusionSperm morphology, sperm DNA fragmentation index and seminal plasma zinc levels may influence the outcome of IVF-ET.The above parameters can be used to predict pregnancy outcome of IVF-ET.

Study on the Relationship between Sperm DNA Fragmentation Index and Hcy Level in Patients with Severe Spermatoqenesis / 现代检验医学杂志

Objective To investigate the relationship between the homocysteine,sperm DNA fragmentation index and sperm counts of male with severe impaired spermatoqenesis.Methods From December 2015 to February 2017,56 male patients with severe impaired spermatoqenesis were enrolled in the study.The patients were divided into two groups according to the WHO criteria:severe oligozoospermia and azoospermia group (n =25) and oligoasthenoteratozoospermia group (n =31),and the control group was a male with no reproductive impairment (n=27).The sperm parameters were analyzed by using the computer automatic semen analyzer,sperm DNA fragmentation index and serum Hcy level were detected by sperm chromatin diffusion method and enzyme colorimetric method.Results The median of Sperm DNA fragmentation index and homocysteine level in control groups were 33％ [95％CI(29.0％ ～34.4％)] and 13.2 μmol/L [95％CI(12.4 μmol/L～14.2 μmol/L)],and in severe spermatogenesis groups in these two indicators were 21％ [95％CI(19.0％ ～24.0％)] and 8.9 mol/L [95％CI(8.4 μmol/L～ 9.4 μmol/L)],respectively.The results of these two items were higher than the control group,the difference was statistically significant (t=6.793～7.543,P=0.000).Sperm survival rate in normal control group and severe spermatogenesis group was 71％ [95％ CI(67.8％ ～75.1％)] and 57％[95％CI(52.3％ ～58.0％)],respectively,and the difference was statistically significant (t=-8.475,P=0.000).Sperm DNA fragmentation index was positively correlated with serum Hcy level and sperm concentration,Passing-Bablok regression analysis was:Y=10.705 +0.053X,Y=21.071+0.286X,and Hcy level was negatively correlated with sperm concentration.Conclusion The increase of Hcy level and sperm DNA fragmentation index may be an importantcause of male with severe impaired spermatoqenesis,but the specific mechanism remains to be further studied.

ICSI significantly improved the pregnancy rate of patients with a high sperm DNA fragmentation index / 대한생식의학회지

OBJECTIVE: Correlations between semen parameters and sperm DNA fragmentation index (DFI) were investigated to identify characteristics of sperm without DNA damage that could be used in selecting sperm for intracytoplasmic sperm injection (ICSI). Pregnancy outcomes were compared to determine whether in vitro fertilization (IVF) or ICSI is a better choice for patients who have sperm with a high-DFI. METHODS: Semen analysis was carried out in 388 patients who visited our IVF center for the first time to investigate correlations between sperm DFI and semen parameters. In addition, 1,102 IVF cycles in 867 patients were carried out in the present study; 921 cycles in the low-DFI group (DFI <30%) and 181 cycles in the high-DFI group (DFI ≥30%). Both the low- and high-DFI groups were subdivided into IVF and ICSI cycle groups. RESULTS: Sperm DFI showed significant inverse correlations with sperm motility (r=−0.435, p<0.001) and morphology (r=−0.153, p<0.05). Sperm DFI also showed significant correlations with rapid motility (r=−0.436, p<0.001), and the kinetic parameters of average-path velocity (r=−0.403) and linearity (r=−0.412). Although there was no significant difference in the pregnancy rates between IVF (48.6%) and ICSI (44.8%) in the low-DFI group, the pregnancy rate of ICSI cycles (44.8%, p<0.05) was significantly higher than IVF cycles (25.0%) in the high-DFI group. No significant difference was observed in the abortion rates between the low-DFI (52 of 921, 5.6%) and high-DFI groups (7 of 181, 3.8%). CONCLUSION: ICSI is a better choice than IVF for improving the pregnancy outcomes of patients who have sperm with a high DFI.

Tongjingling improves sperm DNA integrity and reduces oxidative stress in the testis of experimental varicocele rats / 中华男科学杂志

Objective@#To explore the protective effect of Tongjingling (TJL) against sperm DNA damage and oxidative stress in the rat model of experimental varicocele (EVC).@*METHODS@#We randomly divided 75 Wistar male rats into five groups of equal number: sham operation, EVC model, high-dose TJL, mid-dose TJL, and low-dose TJL. The EVC model was established in the rats by partial ligation of the left renal vein, followed by 8 weeks of medication from the 4th week after modeling. Then we observed the general status of the rats, detected the sperm DNA fragmentation index (DFI) in the epididymis by sperm chromatin structure assay (SCSA), and measured the content of hydroperoxide (H2O2) and the activities of catalase (CAT) and superoxide dismutase (SOD) in the testis by colorimetry.@*RESULTS@#Compared with the sham operation group, the EVC models showed significantly increased sperm DFI in the epididymis (P <0.01) and elevated level of H2O2 and activities of CAT and SOD in the testis (P <0.01). In comparison with the EVC models, the rats of the TJL groups exhibited remarkably reduced sperm DFI and H2O2 content, but increased activities of SOD and CAT.@*CONCLUSIONS@#TJL can improve sperm DNA integrity by increasing the activities of SOD and CAT and reducing the H2O2 level and hence oxidative stress in the testis tissue.

Impacts of Chk1 and Chk2 gene expressions on sperm concentration and motility / 中华男科学杂志

Objective@#To study the correlation of the gene expressions of Chk1 and Chk2 with sperm concentration and motility.@*METHODS@#According to sperm concentration and motility (percentage of progressively motile sperm), we divided 80 semen samples into four groups of equal number: normal control, oligozoospermia (OS), asthenospermia (AS), and oligoasthenozoospermia (OAS). We detected the sperm DNA fragmentation index (DFI) and viability and determined the expressions of Chk1 and Chk2 in the sperm by RT-PCR and Western blot.@*RESULTS@#Statistically significant differences were not found in sperm DFI among the control, OS, AS, and OAS groups (21.24±6.93, 19.67±7.64, 21.52±6.92, and 19.28±11.55, P>0.05), but observed in sperm concentration, progressive motility, and viability between the DFI >30% and DFI ≤30% groups (P<0.01). Compared with the normal control, sperm viability was remarkably decreased in the OS, AS, and OAS groups (［83.48±9.87］% vs ［63.86±9.16］%, ［50.45±16.99］%, and ［39.21±15.74］%, P<0.05). RT-PCR showed remarkable differences among the control, OS, AS, and OAS groups in the relative expression level of Chk1 mRNA (0.73±0.22, 0.62±0.14, 1.03±0.39, and 0.92±0.071, P<0.01), which was correlated positively with sperm concentration (b = 80.661, P<0.01) but negatively with sperm motility (b = －19.275, P < 0.01), as well as in that of Chk2 mRNA (0.66±0.30, 0.27±0.09, 0.59±0.19, and 0.42 ± 0.11, P<0.01), which was correlated negatively with sperm concentration (b = －90.809, P<0.01) but positively with sperm motility (b = 27.507, P <0.01). The relative expression levels of the Chk1 protein were significantly different among the four groups (0.63±0.05, 0.42±0.03, 1.13±0.08, and 0.87±0.07, P<0.01), which was correlated positively with sperm concentration (b = 55.74, P<0.01) but negatively with sperm motility (b =－22.649, P<0.01), and so were those of the Chk2 protein (1.23±0.36, 0.37±0.16, 0.87±0.08, and 0.68±0.12, P<0.01), which was correlated negatively with sperm concentration (b =－53.001, P<0.01) but positively with sperm motility (b = 16.676, P < 0.01).@*CONCLUSIONS@#Chk1 and Chk2 are significantly expressed in human sperm. In case of sperm DNA damage, up-regulated Chk1 expression may enhance sperm apoptosis and lead to asthenospermia, while increased Chk2 expression may inhibit spermatogenesis and result in oligospermia.

Effect of sperm spreparation in male infertility on sperm morphology and DNA integrity / 实用医学杂志

Objective To investigate the effect of sperm preparation in male infertility on sperm morphology and DNA fragmentation index(DFI). Methods Four hundred men were divided into the groups of fertile , teratozoospermics and unexplained subfertility. Sperm morphology and DFI were analyzed before and after the sperm preparation. Results Sperm abnormal morphology and DFI in the infertility group were higher than those in the fertile group ,but significantly decreased after sperm preparation. The method of density gradient separation results in obtaining better sperm(P<0.01). Conclusions Compared to the method of swimming up ,the method of density gradient separation could result in obtaining sperm with improved normal morphology and DNA integrity.

Correlation of the DNA fragmentation index and malformation rate of optimized sperm with embryonic development and early spontaneous abortion in IVF-ET / 中华男科学杂志

<p><b>Objective</b>To investigate the effects of the DNA fragmentation index (DFI) and malformation rate (SMR) of optimized sperm on embryonic development and early spontaneous abortion in conventional in vitro fertilization and embryo transfer (IVF-ET).</p><p><b>METHODS</b>We selected 602 cycles of conventional IVF-ET for pure oviductal infertility that had achieved clinical pregnancies, including 505 cycles with ongoing pregnancy and 97 cycles with early spontaneous abortion. On the day of ovum retrieval, we examined the DNA integrity and morphology of the rest of the optimized sperm using the SCD and Diff-Quik methods, established the joint predictor (JP) by logistic equation, and assessed the value of DFI and SMR in predicting early spontaneous abortion using the ROC curve.</p><p><b>RESULTS</b>The DFI, SMR, and high-quality embryo rate were (15.91±3.69)%, (82.85±10.24)%, and 46.53% (342/735) in the early spontaneous abortion group and (9.30±4.22)%, (77.32±9.19)%, and 56.43% (2263/4010) respectively in the ongoing pregnancy group, all with statistically significant differences between the two groups (P<0.05 ). Both the DFI and SMR were the risk factors of early spontaneous abortion (OR = 5.96 and 1.66; both P< 0.01). The areas under the ROC curve for DFI, SMR and JP were 0.893±0.019, 0.685±0.028, and 0.898±0.018, respectively. According to the Youden index, the optimal cut-off values of the DFI and SMR obtained for the prediction of early spontaneous abortion were approximately 15% and 80%. The DFI was correlated positively with SMR (r= 0.31, P<0.01) but the high-quality embryo rate negatively with both the DFI (r= －0.45, P<0.01) and SMR (r= －0.22, P<0.01).</p><p><b>CONCLUSIONS</b>The DFI and SMR of optimized sperm are closely associated with embryonic development in IVF. The DFI has a certain value for predicting early spontaneous abortion with a threshold of approximately 15%, but SMR may have a lower predictive value.</p>

Correlation of oxidative stress with sperm DNA integrity and semen parameters in infertile men with varicocele / 中华男科学杂志

<p><b>Objective</b>To investigate the relationship of oxidative stress with DNA integrity and semen parameters in infertile men with varicocele (VC).</p><p><b>METHODS</b>This prospective study included 98 infertile males with VC. According to the levels of reactive oxygen species (ROS) in the semen, we divided the patients into a high ROS group (n=44) and a low ROS group (n=54), determined the sperm DNA fragmentation index (DFI), motility and morphology, and analyzed their correlation with ROS in the two groups of patients.</p><p><b>RESULTS</b>Compared with the patients of the low ROS group, those of the high ROS group showed a significantly higher DFI (27.38±8.10 vs 34.49±6.05, P=0.039) and a higher concentration of seminal leukocytes (［0.65±0.15］×10⁶/ml vs ［0.86±0.41］×10⁶/ml, P=0.022), but lower sperm motility (［36.16±22.83］% vs ［18.22±25.21］%, P=0.017), percentage of progressively motile sperm (［23.34±11.53］% vs ［16.34±9.22］%, P=0.041), sperm curvilinear velocity (［27.03±6.21］ vs ［20.62±4.38］ μm/s, P=0.013), and sperm linearity (［29.75±8.24］% vs ［18.30±7.93］%, P=0.024). Spearman correlation analysis indicated that the ROS level was correlated positively with the concentration of seminal leukocytes (r=0.41, P<0.01) and DFI (r=0.21, P=0.006), but negatively with sperm curvilinear velocity (r=－0.24, P=0.017), linearity (r=－0.24, P=0.021), motility (r=－0.31, P=0.002), and the percentage of progressively motile sperm (r=－0.41, P=0.012). Additionally, the sperm DFI manifested a significant negative correlation with sperm motility (r=－0.29, P<0.01) and the percentage of progressively motile sperm (r=－0.34, P<0.01).</p><p><b>CONCLUSIONS</b>The level of seminal ROS is positively correlated with the sperm DFI in infertile men with varicocele, and both the ROS level and DNA integrity are associated with semen parameters.</p>

Efficient isolation of sperm with high DNA integrity and stable chromatin packaging by a combination of density-gradient centrifugation and magnetic-activated cell sorting / 대한생식의학회지

OBJECTIVE: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. METHODS: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. RESULTS: The sperm DFI showed a significant correlation (r=–0.347, p<0.001) with sperm motility and morphology (r=–0.114, p<0.05) but not with other semen parameters. The DFI (11.5%±2.0%) of semen samples was significantly reduced by DGC (8.1%±4.1%) or MACS alone (7.4%±3.9%) (p<0.05). The DFI was significantly further reduced by a combination of DGC and MACS (4.1%±1.3%, p<0.05). Moreover, the combination of DGC and MACS (1.6%±1.1%, p<0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC (4.4%±3.2%) or MACS alone (3.4%±2.2%). CONCLUSION: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.