Magnetic-activated cell sorting of nonapoptotic spermatozoa with a high DNA fragmentation index improves the live birth rate and decreases transfer cycles of IVF/ICSI

The present study aimed to evaluate the clinical outcomes of magnetic-activated cell sorting(MACS)in sperm preparation for male subjects with a sperm DNA fragmentation index(DFI)≥30%.A total of 86 patients who had undergone their first long-term long protocol were selected.The protocol involved in vitro fertilization(IVF)and intracytoplasmic sperm injection(ICSI)cycles,and the patients were divided into the MACS or control groups.The MACS group included sperm samples analyzed with MACS that were combined with density gradient centrifugation(DGC)and the swim-up(SU)technique(n=39),and the control group included sperm samples prepared using standard techniques(DGC and SU;n=41).No differences were noted with regard to basic clinical characteristics,number of oocytes retrieved,normal fertilization rate,cleavage rate,or transplantable embryo rate between the two groups in IVF/ICSI.In addition,the clinical pregnancy and implantation rates of the first embryo transfer cycles indicated no significant differences between the two groups.However,there was a tendency to improve the live birth rate(LBR)of the first embryo transfer cycle(63.2%vs 53.9%)and the cumulative LBR(79.5%vs 70.7%)in the MACS group compared with the control group.Moreover,the number of transferred embryos(mean±standard deviation[s.d.]:1.7±0.7 vs 2.3±1.6)and the transfer number of each retrieved cycle(mean±s.d.:1.2±0.5 vs 1.6±0.8)were significantly lower in the MACS group than those in the control group.Thus,the selection of nonapoptotic spermatozoa by MACS for higher sperm DFI could improve assisted reproductive clinical outcomes.

Random sperm DNA fragmentation index is not associated with clinical outcomes in day-3 frozen embryo transfer

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.

Changes in human sperm motility and DNA fragmentation index after incubation at different temperatures following density gradient centrifugation and swim-up procedures

Objective:The purpose of this study was to evaluate the sperm motility and DNA integrity at different temperatures to analyze whether the sperms are suitable on the second day for insemination of in vitro matured oocytes by intra-cytoplasmic sperm injection(ICSI)following density gradient centrifugation(DGC)and swim-up(SU)procedures.Methods:Semen samples were collected from 30 outpatients who visited the Center for Reproductive Medicine for semen analyses.Following sperm selection by DGC and SU procedures,the liquified semen samples were divided into three groups and incubated at 4,25,and 37°C,respectively.Following incubation for 24,48,and 72 hours,the sperm motility and sperm DNA fragmentation index(DFI)were analyzed.Results:Following the combination of DGC and SU procedures,the sperm motility(91.8%±8.6%vs.50.8%±13.1%)and DFI(5.1%±7.9%vs.13.0%±11.6%)were significantly improved(P<0.01)compared to those without any treatment.The sperm motility of the 3 groups significantly declined(P<0.05)post-incubation compared to that of the groups prior incubation.However,sperm motility significantly increased(76.9%±10.4%)(P<0.05)at 25°C compared to that of the other 2 groups(53.5%±11.0%and 47.6%±10.2%).Sperm DFI significantly increased(P<0.05)at 37°C following incubation for 24 and 72 hours in comparison to that of the other 2 groups.However,the sperm DFI did not significantly increase when the sperm samples were incubated at 4(5.7%±5.9%)and 25°C(6.8%±5.6%)for 24 hours compared to that before incubation(5.1%±7.9%).Conclusions:These results indicate that the sperm quality,in terms of motility and DFI,can be efficiently improved by DGC in combination with SU.Following which,the sperm samples can be incubated at 25°C and be used on the second day for insemination of in vitro matured oocytes by ICSI.

Human circBOULE RNAs as potential biomarkers for sperm quality and male infertility

Reliable molecular biomarkers to predict fertility remain scarce.The current study investigated the potential of testis-specific circBOULE RNAs as biomarkers for male infertility and sperm quality.Using reverse transcription-PCR and real-time reverse transcription-PCR assays,we identified seven circular RNAs from the human BOULE gene in human sperm.We observed that the expression level of circEx3-6 was significantly reduced in asthenozoospermia,while the expression levels of both circEx2-6 and circEx2-7 were decreased in terato-zoospermia,compared with the controls.Furthermore,we demonstrated that the expression level of circEx2-6 was negatively correlated with the sperm DNA fragmentation index,and the expression level of circEx2-7 was correlated with both fertilization and cleavage rates in those treated with the assisted reproductive technologies.Further functional analyses in a transgenic fly model supported the roles of circBOULE RNAs in sperm development and human male fertility.Collectively,our findings support that sperm circBOULE RNAs may serve as diagnostic biomarkers for assessing sperm motility and DNA quality.Therefore,clinical application and significance of sperm circBOULE RNAs in the assisted reproductive technologies warrant further investigation.

Landscape pattern and fragmentation of natural secondary forests in the eastern mountainous region, northeast China： A case study of Mao＇ershan forests in Heilongjiang Province

Mao'ershan region is a representative natural secondary forested region in the eastern mountainous region, northeast of China.. Under the support of ARC/INFO and GIS technology, the landscape shape and fragment indices of Mao'ershan experimental plantation were studied by combining the forest type map (1:10000), which was drawn from the aerial photographs (1999), field investigation (1999) and soil utilized map (1:10000). The results showed that the shape index and shape fragment index of natural landscape were higher than those of artificial landscapes and landscape patch fragment index depended on the number of patches. The natural forest had complex shape, suffering little jamming, and its shape index was higher than that of artificial forest. The manual controlled landscape (e.g. nursery, cropland and cutting blank) had regular shape, and its shape index was smaller. The fragment index of patches in natural forest was higher than that of artificial forest. The soft broad-leaved had the highest fragment index of patch amount.

Effect of Sperm Preservation Solution of Pseudobagrus hwanghoensis in Artificial Reproduction Based on SCSA

The quality of fish sperm is an important factor affecting artificial reproduction.Since the sexual maturation of male and female is not synchronized or for the fish that need to kill the male fish for sperm,sperm preservation solution is used to preserve the sperm temporarily during reproduction,in order to meet the needs of large-scale reproduction.Therefore,the preservation effect of sperm preservation solution directly affects the artificial reproduction.The quality of sperm in the preservation solution can be judged macroscopically by microscopic examination,but it is not accurate,while sperm chromatin structure analysis(SCSA)can quantitatively judge the quality of sperm in the preservation solution,thus providing guarantee for reproduction.The Zhaos sperm preservation solution of Pseudobagrus hwanghoensis has been verified by artificial breeding and SCSA analysis for many times,and it is better than Hanks sperm preservation solution.When the DNA fragmentation index is less than or equal to 0.15,the sperm is vigorous and the effect is obvious.

Decreased AKAP4/PKA signaling pathway in high DFI sperm affects sperm capacitation

The sperm DNA fragmentation index(DFI)is a metric used to assess DNA fragmentation within sperm.During in vitro fertilizationembryo transfer(IVF-ET),high sperm DFI can lead to a low fertilization rate,poor embryo development,early miscarriage,etc.A kinase anchoring protein(AKAP)is a scaffold protein that can bind protein kinase A(PKA)to subcellular sites of specific substrates and protects the biophosphorylation reaction.Sperm protein antigen 17(SPA17)can also bind to AKAP.This study intends to explore the reason for the decreased fertilization rate observed in high sperm DFI(H-DFI)patients during IVF-ET.In addition,the study investigates the expression of AKAP,protein kinase A regulatory subunit(PKARIl),and SPA17 between H-DFI and low sperm DFI(L-DFI)patients.SPA17 at the transcriptional level is abnormal,the translational level increases in H-DFI patients,and the expression of AKAP4/PKARIl protein decreases.H,O,has been used to simulate oxidative stress damage to spermatozoa during the formation of sperm DFI.It indicates that H,O,increases the expression of sperm SPA17 protein and suppresses AKAP4/PKARIl protein expression.These processes inhibit sperm capacitation and reduce acrosomal reactions.Embryo culture data and IVF outcomes have been documented.The H-DFI group has a lower fertilization rate.Therefore,the results indicate that the possible causes for the decreased fertilization rate in the H-DFI patients have included loss of sperm AKAP4/PKARIl proteins,blocked sperm capacitation,and reduced occurrence of acrosome reaction.

Sperm DNA fragmentation in Chinese couples with unexplained recurrent pregnancy loss

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 v/tro fertiIization 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 A30.0%)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%)tha n that in the control group(54.7%).Subsequent analysis also dem on strated 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.

The effect of age and abstinence time on semen quality:a retrospective study

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.

The Influence of Sperm DNA Damage and Semen Homocysteine on Male Infertility

Background:To explore the relationship of sperm DNA fragmentation index(DFI),serum and seminal plasma homocysteine(Hcy),and semen parameters in patients with severe spermatogenetic dysfunction.Methods:A total of 77 infertile males treated in our hospital for severe spermatogenetic dysfunction from January 2016 to November 2017 were recruited.The involved patients were divided into two groups:oligozoospermia(SOM group,35 cases)and asthenozoospermia(OAT group,42 cases).The control group(NM group)contained 31 healthy males without reproductive dysfunctions.All the participants involved were tested in the items below:spermatozoa parameters,spermatozoa DFI,serum Hcy level and seminal plasma Hcy level,concentration of seminal plasma malondialdehyde(MDA),and total antioxidant capacity(TAC).Results:Between the SOM group and NM group,there were significantly difference in sperm concentration,motility and vitality,concentration of MDA,and TAC.The spermatozoa DFI and Hcy levels in SOM group were significantly higher than those of the NM group.Sperm DFI was positively correlated with serum Hcy level(r=0.083,P<0.05).Serum Hcy level was negatively correlated with sperm concentration(r=−0.186,P<0.05)and sperm vitality(r=−0.216,P<0.05).The serum Hcy level was not correlated with sperm Hcy level(r=0.103,P>0.05).Conclusions:The elevated Hcy level and spermatozoa DFI may be important factors of the severe spermatogenetic dysfunction,which can be used as semen index to evaluate sperm quality and male fertility.