Sperm function testing, once commonly performed for the infertile couple before employing assisted reproductive technology （ART）, has fallen out of favour in many reproductive medicine centers throughout the world. ...Sperm function testing, once commonly performed for the infertile couple before employing assisted reproductive technology （ART）, has fallen out of favour in many reproductive medicine centers throughout the world. Indeed, the most recent addition of the ＇World Health Organisation （WHO） Laboratory Manual for the Examination and Processing of Hu- man Semen＇ now groups many of these procedures into a section termed Research Procedures. In large part, this reflects the current clinical practice of bypassing the in-depth evaluation of the male partner, while assuming that if a spermatozoon can be found for intracytoplasmic sperm injection （ICSI）, it must be a healthy cell capable of achieving fertilization. Never- theless, sperm function testing can provide valuable clinical insights into defects causing male infertility. Admittedly, in some cases, functional sperm deficiencies can be overcome using an ART. In other cases, couples will be empowered by the knowledge of the cause of their infertility, and for some couples, perhaps even the likelihood of ICSI success （relative to the spermatozoa）. The knowledge allows them to make truly informed reproductive decisions, including （perhaps） the de- cision to seek donor insemination, to adopt or to remain childless. Knowledge of the cause of their infertility may provide closure for couples and a sense of confidence regarding their choice of reproductive treatment.展开更多
Aim: To determine the predictive value of the hypo-osmotic swelling (HOS) test to identify viable, non-motile sperm. Methods: Semen samples from 20 men with severe asthenozoospermia underwent traditional seminal analy...Aim: To determine the predictive value of the hypo-osmotic swelling (HOS) test to identify viable, non-motile sperm. Methods: Semen samples from 20 men with severe asthenozoospermia underwent traditional seminal analysis, eosin-nigrosin (EN) staining and the HOS test. A further EN stain was then performed on a HOS pre-treated aliquot and a total of 2000 further sperm examined. Results: The median sperm density was 5.1 million/mL (IQR 4.3-13.1) and the median motility was 3.0 % (IQR 0-7). Seven samples showed complete asthenozoospermia. Initial EN staining showed 59 % viability (range 48-69) despite the poor standard parameters and 47 % (range 33-61) in the complete asthenozoospermia subgroup. The HOS test showed 49.9 % reacted overall (range 40-59) and 41.7 % (range 22-61) in the complete asthenozoospermia subgroup. The combined HOS/EN stain showed the positive predictive value of the HOS test to identify viable sperm was 84.2 % overall and 79.7 % in the complete asthenozoospermia subgroup. Conclusion: The HOS test can effectively predict sperm viability in patients with severe and complete asthenozoospermia.展开更多
Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects aftervasovasostomy (Group A: 1-6 months, Group B: 6-12 months; Group C: 12-18 months after vasectomy rever-sal) were inv...Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects aftervasovasostomy (Group A: 1-6 months, Group B: 6-12 months; Group C: 12-18 months after vasectomy rever-sal) were investigated. Semen from 34 normal fertile men was used as controls. Sperm function tests, including hypo-osmotic swelling test (HOST), acridine orange (AO) fluorescence, acrosome reaction (triple-stain), cervical mucuspenetration test (CMPT), etc were done. Results: After vasectomy reversal, the percentage of HOST was signifi-cantly lower than that of the normal fertile men. In regard to AO, there were no significant differences between thethree vasovasostomy groups and between these 3 groups and the controls. With triple-stain, the percentage of normalacrosome reaction was significantly lower in Group A as compared with the controls, but not in Groups B and C. Therewere no significant differences in the results of CMPT between the vasovasostomy groups and the controls. However,the number of 'poor' type was significantly higher in Groups A and C than in the controls; the percentage of 'nega-tive' type were higher in Groups A and B than in the controls. Conclusion: After vasovasostomy a lower level ofHOST remained for one year and gradually recovered after one year. Six months after vasectomy reversal, the percent-age of acrosome reaction could be changed from lower level to normal range. The data of AO indicated that the geneticmaterial (double-stranded DNA) in spermatozoa was not affected by vasovasostomy. To evaluate the result of CMPTafter vasectomy reversal, not only the normal results but also the abnormal results ('poor' and 'negative' types)should also be considered.Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects aftervasovasostomy (Group A: 1-6 months, Group B: 6-12 months; Group C: 12-18 months after vasectomy rever-sal) were investigated. Semen from 34 normal fertile men was used as controls. Sperm function tests, including hypo-osmotic swelling test (HOST), acridine orange (AO) fluorescence, acrosome reaction (triple-stain), cervical mucuspenetration test (CMPT), etc were done. Results: After vasectomy reversal, the percentage of HOST was signifi-cantly lower than that of the normal fertile men. In regard to AO, there were no significant differences between thethree vasovasostomy groups and between these 3 groups and the controls. With triple-stain, the percentage of normalacrosome reaction was significantly lower in Group A as compared with the controls, but not in Groups B and C. Therewere no significant differences in the results of CMPT between the vasovasostomy groups and the controls. However,the number of 'poor' type was significantly higher in Groups A and C than in the controls; the percentage of 'nega-tive' type were higher in Groups A and B than in the controls. Conclusion: After vasovasostomy a lower level ofHOST remained for one year and gradually recovered after one year. Six months after vasectomy reversal, the percent-age of acrosome reaction could be changed from lower level to normal range. The data of AO indicated that the geneticmaterial (double-stranded DNA) in spermatozoa was not affected by vasovasostomy. To evaluate the result of CMPTafter vasectomy reversal, not only the normal results but also the abnormal results ('poor' and 'negative' types)should also be considered.Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects aftervasovasostomy (Group A: 1-6 months, Group B: 6-12 months; Group C: 12-18 months after vasectomy rever-sal) were investigated. Semen from 34 normal fertile men was used as controls. Sperm function tests, including hypo-osmotic swelling test (HOST), acridine orange (AO) fluorescence, acrosome reaction (triple-stain), cervical mucuspenetration test (CMPT), etc were done. Results: After vasectomy reversal, the percentage of HOST was signifi-cantly lower than that of the normal fertile men. In regard to AO, there were no significant differences between thethree vasov展开更多
This was a cohort study of in vitro fertilization(IVF)subjects at the University of Utah,Salt Lake City(UT,USA)utilizing partner sperm.Cycles where both the hamster egg penetration test(HEPT)and semen analysis were pe...This was a cohort study of in vitro fertilization(IVF)subjects at the University of Utah,Salt Lake City(UT,USA)utilizing partner sperm.Cycles where both the hamster egg penetration test(HEPT)and semen analysis were performed within 2 years prior to IVF cycles were stratified into four groups based on a normal or an abnormal HEPT and morphology.The mean conventional and intracytoplasmic sperm injection(ICSI)fertilization rates were calculated in each group.We performed a univariate analysis on the primary outcome comparing clinically interesting subjects.We performed a cost-effectiveness analysis of a policy of HEPT versus universal ICSI in couples with an abnormal morphology.Among patients with a normal HEPT,there was no difference in the mean conventional fertilization rates between those with a normal and an abnormal morphology.There was no difference in the mean conventional fertilization rates between subjects with a normal morphology without a hamster test and those with a normal HEPT without a morphology assessment.In 1000 simulated cycles with an abnormal morphology,a policy of HEPT was cost saving compared to universal ICSI,yet produced similar fertilization rates.The HEPT is similar to the World Health Organization edition 5(WHO-5)morphology in predicting successful conventional fertilization while allowing decreased utilization of ICSI.A policy of HEPT for males with abnormal morphology saves cost in selecting couples for a fertilization method.展开更多
The study was planned with an objective to assess the level of antisperm antibodies (ASA) in the blood serum and seminal plasma of breeding cow bulls and their relationship with sperm function and fertility tests. ASA...The study was planned with an objective to assess the level of antisperm antibodies (ASA) in the blood serum and seminal plasma of breeding cow bulls and their relationship with sperm function and fertility tests. ASA was analyzed in blood serum and seminal plasma by SpermMar test, Immuno peroxidase assay (IPA) and Enzyme linked immunoabsorbant assay (ELISA). In SpermMar test, about 54% bulls were with >40% IgG in blood serum against sperm surface antigens, whereas none of the bulls were with >10% IgG in seminal plasma. More than 20% and >10% IgA against sperm surface antigens were detected in the blood serum and seminal plasma of 65.8% and 37% bulls, respectively. Out of 26 bulls, seminal plasma of 21 bulls reacted with spermatozoa both in IPA and IgA latex particles and that of only 12 bulls reacted with IgG. In IPA, about 50% of the bulls had >40% ASA against head surface antigens, whereas, there were 23% bulls with >10% ASA in seminal plasma. Also ELISA indicated a higher antibody titre in blood serum (3200 - 6400) and seminal plasma (40 - 80) of 50% and 42% bulls, respectively. There were 11 bulls with low values of HOST/in vitro acrosome reaction/cervical mucus penetration assay and higher level of either serum or seminal plasma ASA. Our study revealed that a significant level of ASA in serum or seminal plasma may have effect on the fertility of bulls by affecting the sperm function.展开更多
It is well known that transit through the epididymis involves an increase in the compaction of sperm chromatin, which acquires fully condensed status at the caput epididymidis. The purpose of this study was to compare...It is well known that transit through the epididymis involves an increase in the compaction of sperm chromatin, which acquires fully condensed status at the caput epididymidis. The purpose of this study was to compare the terminal deoxyribonucleotidyl transferase-mediated dUTP nick end-labelling （TUNEL） assay, the comet assay, the sperm chromatin structure assay （SCSA） and the sperm chromatin dispersion （SCD） test by analysing spermatozoa from the caput and cauda epididymidis in order to demonstrate the ability of each technique to discriminate between different degrees of sperm maturity related to chromatin compaction and DNA fragmentation. Our results suggest that some populations of DNA-fragmented spermatozoa associated with immature sperm can only be identified using the comet assay and the SCSA but not with the SCD test or the TUNEL assay.展开更多
Aim： To study the influence of enterococci on human sperm membrane in vitro. Methods： Ejaculated human sperm were artificially infected with β-hemolytic or non-β-hemolytic enterococci at the bacteria： sperm ratio...Aim： To study the influence of enterococci on human sperm membrane in vitro. Methods： Ejaculated human sperm were artificially infected with β-hemolytic or non-β-hemolytic enterococci at the bacteria： sperm ratio of 50：1 at 37℃. Sperm membrane integrity was examined after incubation for 1, 3 and 5 h by hypoosmotic swelling （HOS） test and electron microscopy. Results： Sperm infected with β-hemolytic enterococci had lower HOS scores compared with non-β-hemolytic strains or uninfected control （P 〈 0.01）. The HOS test scores of sperm infected with β-hemolytic enterococci increased in the presence of phosphatidylcholine, an inhibitor of hemolysin. Non-β-hemolytic strains showed no significant difference in swelling rate, compared with the control group （P 〉 0.05）. It was shown by electron microscopy that β-hemolytic enterococci caused significant rupture of human sperm membrane. Conclusion： β-hemolytic enterococci caused human sperm membrane injury, and might be mediated by the hemolysin of enterococci.展开更多
Background:The analysis of chromatin integrity has become an important determinant of sperm quality.In frozenthawed bovine sperm,neither the sequence of post-thaw injury events nor the dynamics of different types of s...Background:The analysis of chromatin integrity has become an important determinant of sperm quality.In frozenthawed bovine sperm,neither the sequence of post-thaw injury events nor the dynamics of different types of sperm DNA breaks are well understood.The aim of the present work was to describe such sperm degradation aftermath focusing on DNA damage dynamics,and to assess if this parameter can predict pregnancy rates in cattle.Results:A total of 75 cryopreserved ejaculates from 25 Holstein bulls were evaluated at two post-thawing periods(0-2 h and 2-4 h),analyzing global and double-stranded DNA damage through alkaline and neutral Comet assays,chromatin deprotamination and decondensation,sperm motility,viability,acrosomal status,and intracellular levels of total ROS,superoxides and calcium.Insemination of 59,605 females was conducted using sperm from the same bulls,thus obtaining the non-return to estrus rates after 90 d(NRR).Results showed an increased rate of double-stranded breaks in the first period(0-2 h:1.29±1.01%/h vs.2-4 h:0.13±1.37%/h;P<0.01),whereas the rate of sperm with moderate+high single-stranded breaks was higher in the second period(0-2 h:3.52±7.77%/h vs.2-4h:21.06±11.69%/h;P<0.0001).Regarding sperm physiology,viability decrease rate was different between the two periods(0-2 h:-4.49±1.79%/h vs.2-4 h:-2.50±3.39%/h;P=0.032),but the progressive motility decrease rate was constant throughout post-thawing incubation(0-2 h:-4.70±3.42%/h vs.2-4 h:-1.89±2.97%/h;P>0.05).Finally,whereas no correlations between bull fertility and any dynamic parameter were found,there were correlations between the NRR and the basal percentage of highly-damaged sperm assessed with the alkaline Comet(Rs=-0.563,P=0.003),between NRR and basal progressive motility(Rs=0.511,P=0.009),and between NRR and sperm with high ROS at 4 h post-thaw(Rs=0.564,P=0.003).Conclusion:The statistically significant correlations found between intracellular ROS,sperm viability,sperm motility,DNA damage and chromatin deprotamination suggested a sequence of events all driven by oxidative stress,where viability and motility would be affected first and sperm chromatin would be altered at a later stage,thus suggesting that bovine sperm should be used for fertilization within 2 h post-thaw.Fertility correlations supported that the assessment of global DNA damage through the Comet assay may help predict bull fertility.展开更多
Objective: To analyze the application value of combined detection of sperm quality, sex hormone and ovulation in the diagnosis of infertility. Methods: The study was conducted from June 2021 to June 2022. Sixty infert...Objective: To analyze the application value of combined detection of sperm quality, sex hormone and ovulation in the diagnosis of infertility. Methods: The study was conducted from June 2021 to June 2022. Sixty infertile couples who received IVF cycle treatment in our hospital during this period were selected as the observation group, and 60 couples with good sperm quality and follicle number ≥ 5 who conceived naturally after gynecological disease treatment were selected as the control group during the same period. The sperm quality, sex hormones and ovulation of the two groups were observed, and the change of positive rate was detected by the combined detection method. Results: Compared with the control group, the observation group had less semen (2.82 ± 0.12) ml, lower concentration (69.17 ± 1.28) × 106/ml, normal sperm morphology rate (2.92% ± 0.11%), lower survival rate (70.25% ± 1.16%), higher deformed sperm index (1.39 ± 0.11), and significant differences between groups (P < 0.05);The levels of FSH (7.15 ± 1.33) U/L, LH (5.13 ± 0.53) mU/ml, E2 (72.34 ± 5.11) ng/L, AMH (3.87 ± 0.67) ng/ml and AFC (7.15 ± 0.76) in the control group were significantly better than those in the observation group (P < 0.05). Compared with the single detection method of the three groups, the positive detection rate of the combined diagnosis method was higher, and the difference between the groups was significant (P < 0.05). Conclusion: For the diagnosis of infertility, the combined detection method of sperm quality, sex hormone and ovulation can effectively clarify the problems existing in the couple, significantly improve the detection rate of the cause of the patient, and is more conducive to guiding clinical symptomatic treatment, which is worthy of promotion and reference.展开更多
文摘Sperm function testing, once commonly performed for the infertile couple before employing assisted reproductive technology （ART）, has fallen out of favour in many reproductive medicine centers throughout the world. Indeed, the most recent addition of the ＇World Health Organisation （WHO） Laboratory Manual for the Examination and Processing of Hu- man Semen＇ now groups many of these procedures into a section termed Research Procedures. In large part, this reflects the current clinical practice of bypassing the in-depth evaluation of the male partner, while assuming that if a spermatozoon can be found for intracytoplasmic sperm injection （ICSI）, it must be a healthy cell capable of achieving fertilization. Never- theless, sperm function testing can provide valuable clinical insights into defects causing male infertility. Admittedly, in some cases, functional sperm deficiencies can be overcome using an ART. In other cases, couples will be empowered by the knowledge of the cause of their infertility, and for some couples, perhaps even the likelihood of ICSI success （relative to the spermatozoa）. The knowledge allows them to make truly informed reproductive decisions, including （perhaps） the de- cision to seek donor insemination, to adopt or to remain childless. Knowledge of the cause of their infertility may provide closure for couples and a sense of confidence regarding their choice of reproductive treatment.
文摘Aim: To determine the predictive value of the hypo-osmotic swelling (HOS) test to identify viable, non-motile sperm. Methods: Semen samples from 20 men with severe asthenozoospermia underwent traditional seminal analysis, eosin-nigrosin (EN) staining and the HOS test. A further EN stain was then performed on a HOS pre-treated aliquot and a total of 2000 further sperm examined. Results: The median sperm density was 5.1 million/mL (IQR 4.3-13.1) and the median motility was 3.0 % (IQR 0-7). Seven samples showed complete asthenozoospermia. Initial EN staining showed 59 % viability (range 48-69) despite the poor standard parameters and 47 % (range 33-61) in the complete asthenozoospermia subgroup. The HOS test showed 49.9 % reacted overall (range 40-59) and 41.7 % (range 22-61) in the complete asthenozoospermia subgroup. The combined HOS/EN stain showed the positive predictive value of the HOS test to identify viable sperm was 84.2 % overall and 79.7 % in the complete asthenozoospermia subgroup. Conclusion: The HOS test can effectively predict sperm viability in patients with severe and complete asthenozoospermia.
文摘Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects aftervasovasostomy (Group A: 1-6 months, Group B: 6-12 months; Group C: 12-18 months after vasectomy rever-sal) were investigated. Semen from 34 normal fertile men was used as controls. Sperm function tests, including hypo-osmotic swelling test (HOST), acridine orange (AO) fluorescence, acrosome reaction (triple-stain), cervical mucuspenetration test (CMPT), etc were done. Results: After vasectomy reversal, the percentage of HOST was signifi-cantly lower than that of the normal fertile men. In regard to AO, there were no significant differences between thethree vasovasostomy groups and between these 3 groups and the controls. With triple-stain, the percentage of normalacrosome reaction was significantly lower in Group A as compared with the controls, but not in Groups B and C. Therewere no significant differences in the results of CMPT between the vasovasostomy groups and the controls. However,the number of 'poor' type was significantly higher in Groups A and C than in the controls; the percentage of 'nega-tive' type were higher in Groups A and B than in the controls. Conclusion: After vasovasostomy a lower level ofHOST remained for one year and gradually recovered after one year. Six months after vasectomy reversal, the percent-age of acrosome reaction could be changed from lower level to normal range. The data of AO indicated that the geneticmaterial (double-stranded DNA) in spermatozoa was not affected by vasovasostomy. To evaluate the result of CMPTafter vasectomy reversal, not only the normal results but also the abnormal results ('poor' and 'negative' types)should also be considered.Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects aftervasovasostomy (Group A: 1-6 months, Group B: 6-12 months; Group C: 12-18 months after vasectomy rever-sal) were investigated. Semen from 34 normal fertile men was used as controls. Sperm function tests, including hypo-osmotic swelling test (HOST), acridine orange (AO) fluorescence, acrosome reaction (triple-stain), cervical mucuspenetration test (CMPT), etc were done. Results: After vasectomy reversal, the percentage of HOST was signifi-cantly lower than that of the normal fertile men. In regard to AO, there were no significant differences between thethree vasovasostomy groups and between these 3 groups and the controls. With triple-stain, the percentage of normalacrosome reaction was significantly lower in Group A as compared with the controls, but not in Groups B and C. Therewere no significant differences in the results of CMPT between the vasovasostomy groups and the controls. However,the number of 'poor' type was significantly higher in Groups A and C than in the controls; the percentage of 'nega-tive' type were higher in Groups A and B than in the controls. Conclusion: After vasovasostomy a lower level ofHOST remained for one year and gradually recovered after one year. Six months after vasectomy reversal, the percent-age of acrosome reaction could be changed from lower level to normal range. The data of AO indicated that the geneticmaterial (double-stranded DNA) in spermatozoa was not affected by vasovasostomy. To evaluate the result of CMPTafter vasectomy reversal, not only the normal results but also the abnormal results ('poor' and 'negative' types)should also be considered.Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects aftervasovasostomy (Group A: 1-6 months, Group B: 6-12 months; Group C: 12-18 months after vasectomy rever-sal) were investigated. Semen from 34 normal fertile men was used as controls. Sperm function tests, including hypo-osmotic swelling test (HOST), acridine orange (AO) fluorescence, acrosome reaction (triple-stain), cervical mucuspenetration test (CMPT), etc were done. Results: After vasectomy reversal, the percentage of HOST was signifi-cantly lower than that of the normal fertile men. In regard to AO, there were no significant differences between thethree vasov
文摘This was a cohort study of in vitro fertilization(IVF)subjects at the University of Utah,Salt Lake City(UT,USA)utilizing partner sperm.Cycles where both the hamster egg penetration test(HEPT)and semen analysis were performed within 2 years prior to IVF cycles were stratified into four groups based on a normal or an abnormal HEPT and morphology.The mean conventional and intracytoplasmic sperm injection(ICSI)fertilization rates were calculated in each group.We performed a univariate analysis on the primary outcome comparing clinically interesting subjects.We performed a cost-effectiveness analysis of a policy of HEPT versus universal ICSI in couples with an abnormal morphology.Among patients with a normal HEPT,there was no difference in the mean conventional fertilization rates between those with a normal and an abnormal morphology.There was no difference in the mean conventional fertilization rates between subjects with a normal morphology without a hamster test and those with a normal HEPT without a morphology assessment.In 1000 simulated cycles with an abnormal morphology,a policy of HEPT was cost saving compared to universal ICSI,yet produced similar fertilization rates.The HEPT is similar to the World Health Organization edition 5(WHO-5)morphology in predicting successful conventional fertilization while allowing decreased utilization of ICSI.A policy of HEPT for males with abnormal morphology saves cost in selecting couples for a fertilization method.
文摘The study was planned with an objective to assess the level of antisperm antibodies (ASA) in the blood serum and seminal plasma of breeding cow bulls and their relationship with sperm function and fertility tests. ASA was analyzed in blood serum and seminal plasma by SpermMar test, Immuno peroxidase assay (IPA) and Enzyme linked immunoabsorbant assay (ELISA). In SpermMar test, about 54% bulls were with >40% IgG in blood serum against sperm surface antigens, whereas none of the bulls were with >10% IgG in seminal plasma. More than 20% and >10% IgA against sperm surface antigens were detected in the blood serum and seminal plasma of 65.8% and 37% bulls, respectively. Out of 26 bulls, seminal plasma of 21 bulls reacted with spermatozoa both in IPA and IgA latex particles and that of only 12 bulls reacted with IgG. In IPA, about 50% of the bulls had >40% ASA against head surface antigens, whereas, there were 23% bulls with >10% ASA in seminal plasma. Also ELISA indicated a higher antibody titre in blood serum (3200 - 6400) and seminal plasma (40 - 80) of 50% and 42% bulls, respectively. There were 11 bulls with low values of HOST/in vitro acrosome reaction/cervical mucus penetration assay and higher level of either serum or seminal plasma ASA. Our study revealed that a significant level of ASA in serum or seminal plasma may have effect on the fertility of bulls by affecting the sperm function.
文摘It is well known that transit through the epididymis involves an increase in the compaction of sperm chromatin, which acquires fully condensed status at the caput epididymidis. The purpose of this study was to compare the terminal deoxyribonucleotidyl transferase-mediated dUTP nick end-labelling （TUNEL） assay, the comet assay, the sperm chromatin structure assay （SCSA） and the sperm chromatin dispersion （SCD） test by analysing spermatozoa from the caput and cauda epididymidis in order to demonstrate the ability of each technique to discriminate between different degrees of sperm maturity related to chromatin compaction and DNA fragmentation. Our results suggest that some populations of DNA-fragmented spermatozoa associated with immature sperm can only be identified using the comet assay and the SCSA but not with the SCD test or the TUNEL assay.
基金Acknowledgment This work was supported by grants from Natural Science Foundation of Fujian Province, China （No. C0510008） and Science ＆ Technology Development Plan of Fujian Province, China （No. C0101080）.
文摘Aim： To study the influence of enterococci on human sperm membrane in vitro. Methods： Ejaculated human sperm were artificially infected with β-hemolytic or non-β-hemolytic enterococci at the bacteria： sperm ratio of 50：1 at 37℃. Sperm membrane integrity was examined after incubation for 1, 3 and 5 h by hypoosmotic swelling （HOS） test and electron microscopy. Results： Sperm infected with β-hemolytic enterococci had lower HOS scores compared with non-β-hemolytic strains or uninfected control （P 〈 0.01）. The HOS test scores of sperm infected with β-hemolytic enterococci increased in the presence of phosphatidylcholine, an inhibitor of hemolysin. Non-β-hemolytic strains showed no significant difference in swelling rate, compared with the control group （P 〉 0.05）. It was shown by electron microscopy that β-hemolytic enterococci caused significant rupture of human sperm membrane. Conclusion： β-hemolytic enterococci caused human sperm membrane injury, and might be mediated by the hemolysin of enterococci.
基金the European Union’s Horizon 2020 Research and Innovation scheme under the Marie Sklodowska-Curie grant agreement No.801342(Tecniospring INDUSTRY,TECSPR-19-1-0003)the Ministry of Science and Innovation,Spain(AGL2017-88329-R and PID2020-113320RBI00)+2 种基金the Catalan Agency for Management of University and Research Grants,Regional Government of Catalonia,Spain(2017-SGR-1229)the Catalan Institution for Research and Advanced Studies(ICREA)La Maratóde TV3 Foundation(214/857-202039)。
文摘Background:The analysis of chromatin integrity has become an important determinant of sperm quality.In frozenthawed bovine sperm,neither the sequence of post-thaw injury events nor the dynamics of different types of sperm DNA breaks are well understood.The aim of the present work was to describe such sperm degradation aftermath focusing on DNA damage dynamics,and to assess if this parameter can predict pregnancy rates in cattle.Results:A total of 75 cryopreserved ejaculates from 25 Holstein bulls were evaluated at two post-thawing periods(0-2 h and 2-4 h),analyzing global and double-stranded DNA damage through alkaline and neutral Comet assays,chromatin deprotamination and decondensation,sperm motility,viability,acrosomal status,and intracellular levels of total ROS,superoxides and calcium.Insemination of 59,605 females was conducted using sperm from the same bulls,thus obtaining the non-return to estrus rates after 90 d(NRR).Results showed an increased rate of double-stranded breaks in the first period(0-2 h:1.29±1.01%/h vs.2-4 h:0.13±1.37%/h;P<0.01),whereas the rate of sperm with moderate+high single-stranded breaks was higher in the second period(0-2 h:3.52±7.77%/h vs.2-4h:21.06±11.69%/h;P<0.0001).Regarding sperm physiology,viability decrease rate was different between the two periods(0-2 h:-4.49±1.79%/h vs.2-4 h:-2.50±3.39%/h;P=0.032),but the progressive motility decrease rate was constant throughout post-thawing incubation(0-2 h:-4.70±3.42%/h vs.2-4 h:-1.89±2.97%/h;P>0.05).Finally,whereas no correlations between bull fertility and any dynamic parameter were found,there were correlations between the NRR and the basal percentage of highly-damaged sperm assessed with the alkaline Comet(Rs=-0.563,P=0.003),between NRR and basal progressive motility(Rs=0.511,P=0.009),and between NRR and sperm with high ROS at 4 h post-thaw(Rs=0.564,P=0.003).Conclusion:The statistically significant correlations found between intracellular ROS,sperm viability,sperm motility,DNA damage and chromatin deprotamination suggested a sequence of events all driven by oxidative stress,where viability and motility would be affected first and sperm chromatin would be altered at a later stage,thus suggesting that bovine sperm should be used for fertilization within 2 h post-thaw.Fertility correlations supported that the assessment of global DNA damage through the Comet assay may help predict bull fertility.
文摘Objective: To analyze the application value of combined detection of sperm quality, sex hormone and ovulation in the diagnosis of infertility. Methods: The study was conducted from June 2021 to June 2022. Sixty infertile couples who received IVF cycle treatment in our hospital during this period were selected as the observation group, and 60 couples with good sperm quality and follicle number ≥ 5 who conceived naturally after gynecological disease treatment were selected as the control group during the same period. The sperm quality, sex hormones and ovulation of the two groups were observed, and the change of positive rate was detected by the combined detection method. Results: Compared with the control group, the observation group had less semen (2.82 ± 0.12) ml, lower concentration (69.17 ± 1.28) × 106/ml, normal sperm morphology rate (2.92% ± 0.11%), lower survival rate (70.25% ± 1.16%), higher deformed sperm index (1.39 ± 0.11), and significant differences between groups (P < 0.05);The levels of FSH (7.15 ± 1.33) U/L, LH (5.13 ± 0.53) mU/ml, E2 (72.34 ± 5.11) ng/L, AMH (3.87 ± 0.67) ng/ml and AFC (7.15 ± 0.76) in the control group were significantly better than those in the observation group (P < 0.05). Compared with the single detection method of the three groups, the positive detection rate of the combined diagnosis method was higher, and the difference between the groups was significant (P < 0.05). Conclusion: For the diagnosis of infertility, the combined detection method of sperm quality, sex hormone and ovulation can effectively clarify the problems existing in the couple, significantly improve the detection rate of the cause of the patient, and is more conducive to guiding clinical symptomatic treatment, which is worthy of promotion and reference.
