Consequences of butylated hydroxytoluene in the freezing extender on post-thaw characteristics of stallion spermatozoa in vitro

Ejaculates from six pure Spanish stallions were split, and one subsample frozen in a commercial extender supplemented with the lipid soluble antioxidant butylated hydroxytoluene (BHT), while the other subsample served as control. After at least 4 weeks of storage, samples were thawed and post-thaw sperm quality analysed: sperm motility and kinematics using a CASA system, membrane and acrosome integrity and mitochondrial membrane potential using flow cytometry. The outcome of cryopreservation varied significantly among stallions. However, the supplementation with 1 mm BHT had no significant effect on any of the sperm parameters evaluated post-thaw.     

Effects of centrifugation before freezing on boar sperm cryosurvival

Current protocols for boar sperm cryopreservation require the centrifugation of semen in order to separate sperm cells from the seminal plasma. This study evaluated the influence of different centrifugation regimes on both sperm recovery and yield (percentage of viable sperm with an intact acrosome relative to the initial sperm population) after centrifugation (experiment 1) as well as the influence of different centrifugation regimes on boar sperm cryosurvival (experiment 2). In both experiments, sperm-rich fractions from 3 boars were diluted, pooled, and cooled to 17 degrees C before centrifugation. In experiment 1, the g-forces tested were 400, 800, 1600, and 2400 x g for 3 or 5 minutes, using the standard regime (800 x g for 10 minutes) as a reference. Sperm recovery (Bürker Chamber) and yield (triple fluorescent stain of PI/R123/FITC-PNA [DNA-specific fluorochrome propidium iodide/mitochondria-specific fluorochrome rhodamine-123/acrosome-specific fluorochrome fluorescein isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin]) were calculated. The highest recovery and yield (P <.05) values were achieved using 2400 x g for 5 or 3 minutes and 1600 x g for 5 minutes, which showed no differences (P >.05) from the reference in terms of sperm yield. In experiment 2, cooled semen was centrifuged using 3 different regimes: C1 (2400 x g for 3 minutes), C2 (1600 x g for 5 minutes), and C3 (800 x g for 10 minutes). Pellets were diluted in lactose-egg yolk (LEY)-glycerol-Equex STM (1 x 10(9) cells/mL) and frozen in 0.5-mL straws. After thawing, sperm quality was assessed after 30 and 150 minutes of incubation (37 degrees C). Centrifugation regimes C1 and C2 showed significantly (P <.05) higher postthaw sperm motility (assessed with a computer-assisted semen analysis system), viability (evaluated as for experiment 1), and percentage of uncapacitated sperm (assessed with a chlortetracycline assay) than did C3. In addition, C1 had the highest (P <.05) oocyte penetrating ability (assessed with the homologous in vitro penetration test performed with immature oocytes). Malondialdehyde production, assessed with the thiobarbituric acid reactive species test, was unaffected (P >.05) by the centrifugation regime used. We conclude that high g-force (2400 x g) and short centrifugation time (3 minutes) do not affect sperm recovery and yield and that, moreover, they have a positive effect on the cryosurvival of boar sperm. Therefore, we recommend the use of short-term centrifugation with a relatively high g-force (2400 x g for 3 minutes) in boar sperm cryopreservation protocol.     

Sperm indexes obtained using computer-assisted morphometry provide a forecast of the freezability of canine sperm

Sperm morphometric indexes obtained after principal component analysis were used to investigate its value as diagnostic tests for freezability. Semen from six dogs was frozen-thawed following a standard protocol. Before freezing, computer-assisted analysis of sperm morphometry (CASMA) was performed. The principal component analysis (a statistical technique for simplifying a dataset, by reducing multidimensional datasets to lower dimensions for analysis) performed in the data obtained after the CASMA analysis gave four morphometric indexes. After thawing, ejaculates were evaluated for early changes in sperm membranes using the combination of two fluorescent probes, YO-PRO-1 and ethidium homodimer and flow cytometry. Significant differences in the percentages of intact membranes post-thaw were observed among dogs (p < 0.01). Significant non-parametric correlations were found between index 3 and the percentage of intact membranes after thawing (R = 0.432 p < 0.05). Receiving operating system curves demonstrate a good diagnostic value for indexes 2 and 3 in the prediction of freezability, with areas under the curve of 0.798 and 0.786, respectively.     

不同冷冻方法对人睾丸精子冷冻保存效果的研究

目的比较不同冷冻保护剂和不同冷冻载体对人睾丸精子的冷冻保存效果。方法选取在兰州大学第一医院生殖医学专科医院进行诊断性睾丸穿刺获得成熟活动精子的患者,以及行睾丸精子ICSI术后的患者,在知情同意下将剩余睾丸组织冷冻保存,共46例。将每例睾丸组织分为4份,随机分成4组进行冷冻:1.8ml冷冻管+甘油复合冷冻剂(A组,46例);1.8ml冷冻管+Fasrun冷冻保护剂(B组,46例);0.25 ml麦管+Fasrun冷冻保护剂(C组,46例);超微量冷冻麦管+Fasrun冷冻保护剂(D组,46例)。冷冻标本经液氮熏蒸后投入液氮,比较各组复苏后精子活力和复苏率。结果冷冻前各组的活动睾丸精子能够满足ICSI操作所用精子量;冷冻复苏后,C组和D组能够容易找到正常活动精子(≥0.1/200倍视野)的例数显著高于A组、B组(P0.05);D组解冻后运动精子复苏率显著高于A组、B组(P0.05),C组解冻后运动精子复苏率显著高于A组(P0.05)。结论应用改良的超微量冷冻麦管+Fasrun冷冻保护剂能够提高睾丸精子的冷冻复苏率。     

弗司扣林对人精子运动功能的影响

目的 通过研究弗司扣林(forskolin)对体外人精子的运动功能有无影响，了解环磷酸腺苷／蛋白激酶A(cAMP／PKA)信号传导途径是否参与人精子运动功能的调节。方法 10例健康成年男性手淫获得新鲜精液，经上游优化处理后与不同浓度的forskolin一起孵育20、30、60min后，采用计算机辅助的精子分析系统(CASA)检测精子的各项运动参数，并进行对比分析。结果 forskolin在体外能显著提高人精于的活率及前向性运动百分率，而对精子的形态及直线速度(VSL)和曲线速度(VCL)无明显影响。结论 forskolin在体外能提高人精子的运动功能，此结果为证实cAMP/PKA信号传导通道参与调节人精子运动功能提供了一定的实验依据。     

流式细胞术检测精索静脉曲张患者精子质膜完整性的研究

目的:应用荧光染料SYBR-14/PI双色标记法进行流式细胞术检测精索静脉曲张患者精子质膜的完整性并探讨其临床意义。方法:随机收集120例男性精液标本,其中90例左侧精索静脉曲张患者分为3组:VC1组(精索静脉曲张Ⅰ度,n=30)、VC2组(精索静脉曲张Ⅱ度,n=30)和VC3组(精索静脉曲张Ⅲ度,n=30),正常生育男性为正常对照组(n=30)。通过计算机辅助精液分析系统进行精液常规分析。精液标本经PBS洗涤处理后用荧光染料SYBR-14/碘化丙锭(PI)双染后上流式细胞仪分析,用发绿色荧光精子百分率(SYBR-14+/PI-%)表示质膜完整精子(PMI)的比例,检测精子质膜完整性,并预见精子的受精能力。结果:精索静脉曲张各组患者与正常对照组之间SYBR-14+/PI-%、SYBR-14-/PI+%均存在统计学差异(P<0.01)。精索静脉曲张患者(VC1、VC2、VC3)代表精子质膜完整性指标(SYBR-14+/PI-%)的百分率分别是:[(54.85±3.78)%]、[(45.37±4.12)%]、[(35.14±4.91)%],均显著低于正常对照组[(70.79±6.71)%],其活力顺序为VC30.05)。结论:应用流式细胞术SYBR-14/PI双染法能快速、准确地检测精子质膜完整性,精索静脉曲张引起精子质膜完整性下降,可能是导致男性不育的重要原因之一。     

溶脲脲原体感染对男性不育患者精子质膜完整性的影响

目的:应用荧光染料SYBR-14/碘化丙锭(PI)双标法流式细胞术检测溶脲脲原体(Uu)感染对男性不育患者精子质膜完整性的影响。方法:收集63例男性精液标本,分为Uu感染组(n=32)和正常对照组(n=31)。通过计算机辅助精液分析系统进行精液常规分析,培养法进行Uu检测。精液标本经PBS洗涤处理后用荧光染料SYBR-14/PI双染后上流式细胞仪分析,用发绿色荧光精子百分率(SYBR-14+/PI-%)表示质膜完整精子的比例,检测精子质膜完整性。结果:Uu感染组精子质膜完整性[(45.14±10.69)%versus(72.68±9.91)%]和(a+b)级精子百分率[(23.29±8.81)%versus(46.32±9.54)%]均显著低于正常对照组(P<0.01),两组间精液量、pH值、精子浓度差异无显著性(P>0.05)。结论:Uu感染引起精子质膜完整性下降,可能是导致男性不育的重要原因之一。     

SYBR-14/PI双染法流式细胞术检测精子质膜完整性的研究

目的:探讨应用荧光染料SYBR-14/碘化丙啶(SYBR-14/PI)双色标记法进行流式细胞术检测精子质膜完整性的可行性及其临床意义。方法:收集208例男性精液标本,按WHO精液分析标准分为正常组(n=31)与异常组(n=177)。通过计算机辅助精液分析系统进行精液常规分析。精液标本洗涤处理后经SYBR-14/PI双染后上流式细胞仪(FCM)分析,用发绿色荧光精子百分率(SYBR-14+/PI-%)表示质膜完整精子(PMI)的比例。结果:正常组与异常组SYBR-14+/PI-与SYBR-14-/PI+精子百分率均存在统计学差异(P均<0.05)。正常组精子SYBR-14+/PI-%为(55.66±20.64)%,显著高于异常组[(39.71±19.21)%,P=0.000]。208例标本中,SYBR-14+/PI-%与精子活动率呈显著正相关(r=0.408,P=0.000),与(a+b)级精子百分率呈显著正相关(r=0.398,P=0.000),与d级精子百分率呈显著负相关(r=-0.413,P=0.000);SYBR-14-/PI+%与精子活动率呈显著负相关(r=-0.380,P=0.000),与(a+b)级精子百分率呈显著负相关(r=-0.397,P=0.000),与d级精子百分率呈显著正相关(r=0.385,P=0.000);SYBR-14+/PI+%与精子活动率呈正相关(r=0.172,P=0.013),与(a+b)级精子百分率呈正相关(r=0.177,P=0.011),与d级精子百分率呈负相关(r=-0.164,P=0.018)。结论:应用流式细胞术SYBR-14/PI双染法检测精子质膜完整性具有可行性,可用于评估男性生育力。     

不同年龄对精子凋亡率及DNA完整性影响的研究

目的:探讨不同年龄阶段精子凋亡率、精子DNA完整性及精液基本参数与年龄是否存在相关性。方法:随机选取在我科因女方因素行体外受精-胚胎移植男性精液标本104例,按男方年龄分为3组,年龄<35岁组43例、35～39岁组31例、≥40岁组30例,按WHO手册第4版手工分析精液常规参数,流式细胞术(FCM)结合FITC-Annexin V/PI荧光染色检测精子凋亡率,吖啶橙荧光染色检测精子DNA完整性。结果:①3组精液量[(2.87±0.89)ml vs(2.98±1.09)ml vs(2.65±0.95)ml]、精子浓度[(60.40±25.43)×106/ml vs(69.74±28.33)×106/ml vs(55.97±27.22)×106/ml]无显著性差异(P>0.05),≥40岁组前向运动精子百分率[(39.00±8.35)%]减少,与<35岁组[(48.72±9.89)%]和35～39岁组[(45.65±10.55)%]相比,差异有显著性(P<0.01);≥40岁组正常形态精子百分率[(11.11±8.26)%]与<35岁组[(16.43±8.75)%]相比,差异有显著性(P<0.01)。②精子凋亡在不同年龄组人群均有一定发生率,≥40岁组精子凋亡率为[(11.82±5.77)%],高于<35岁组[(7.04±3.50)%]和35～39岁组[(9.42±4.73)%],与<35岁组相比结果有统计学意义(P<0.01);≥40岁组DNA完整精子百分率降低[(75.52±10.60)%],与<35岁组[(86.55±5.60)%]和35～39岁组[(81.39±8.94)%]相比,差异有显著性(P<0.01);③男性年龄与精子凋亡百分率呈正相关(P<0.01),与前向运动精子、DNA完整精子百分率呈负相关(P<0.01)。结论:男性年龄增加可能导致前向运动精子百分率减少,正常形态精子百分率降低,精子凋亡百分率增加,DNA完整精子百分率性降低,在生育过程中,男性年龄应引起关注。     

A new and simple method to evaluate early membrane changes in frozen-thawed boar spermatozoa

Detection of early changes in the sperm plasma membrane during cryopreservation is of utmost importance when designing freezing protocols and has previously been studied in the pig species using annexin-V detection of phosphatidylserine translocation. In the present study we designed a new assay to detect these changes in boar spermatozoa, based on the slight increase of sperm membrane permeability occurring during the early stages of cryoinjury, using the combination of three fluorescent probes, SNARF-1, YO-PRO-1 and ethidium homodimer. Four ejaculates from five different boars were frozen-thawed and flow cytometrically (FC) evaluated as paired samples. One of the samples was assayed using the annexin-V/propidium iodide staining and the other sample was evaluated using the new triple staining. Using this combination of probes, four sperm subpopulations were easily detected: viable, with stable membranes (SNARF-1 positive cells), and three with compromised membranes, one of YO-PRO-1+/Eth- cells, one ethidium homodimer+ spermatozoa and, finally spermatozoa stained both with YO-PRO-1 and ethidium homodimer (YO-PRO-1+/Eth+). The latter three categories corresponded to dead spermatozoa, but with different degree of membrane damage, being YO-PRO+/Eth- an earlier stage of membrane destabilization, (manifested by an increase in membrane permeability, while still maintaining membrane integrity) than YO-PRO+/Eth+. A method agreement analysis between both methods was performed revealing good agreement, although the percentage of live cells was 9.44% larger for the triple stain than the annexin-V assay. The new assay stained all sperm sub-populations present in the sample, making it especially suitable for both fluorescence microscopy and flow cytometry, facilitating the exclusion of debris and egg-yolk particles when using FC.     

Relationship between sperm quality and chromatin condensation measured by sperm DNA fluorescence using flow cytometry

DNA flow cytometry of sperm from 100 randomly chosen men undergoing fertility investigation revealed a general association between reduced sperm quality, as judged by conventional parameters, and the appearance of sperm with lower degrees of chromatin condensation in the ejaculate as measured by DNA fluorescence intensity. Chromatin hypocondensation, as measured by increased fluorescence, was manifested to different degrees in different samples. In many cases of more extreme sperm pathology, such as oligoasthenoteratozoospermia (OAT), the whole population of spermatozoa appeared to be affected. Significant numbers of hypercondensed spermatozoa were present in both normozoospermic men and men with different degrees of disturbance in sperm quality. All of the different parameters of sperm quality could be correlated significantly with certain of the flow parameters, although not one in particular could be used to predict deviations from the normal flow profile. In several asthenoteratozoospermic men and a small proportion of men with OAT, the DNA profiles were normal, implying that in these cases the disturbance may not be so fundamental. The presence of leucocytes in the ejaculate was associated with a general increase in the preponderance of hypocondensed subpopulations of spermatozoa in men with OAT as well as in normozoospermic subjects, emphasizing the effect of inflammatory conditions in the reproductive tract on sperm quality.     

Effects of sauna on sperm movement characteristics of normal men measured by computer-assisted sperm analysis

The effects of sauna exposure on sperm movement characteristics and other semen parameters were evaluated using computer-assisted sperm analysis (CASA). A significant ( p  < 0.01) decrease in average path velocity (VAP), curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH) was found after exposure to sauna for 2 weeks. The altered parameters returned to their original values within 1 week after cessation of sauna exposure. Mean values for semen volume, sperm count, percentage motility, sperm morphology and sperm penetration assay (SPA) were not statistically different during and after sauna, when compared to the corresponding control values. The results suggest that increasing scrotal temperature by sauna causes a reversible decrease in sperm movement parameters.     

流式细胞术检测吸烟对精子质膜完整性的影响

目的:应用荧光染料SYBR-14/PI双色标记法进行流式细胞术检测不育患者精子质膜完整性,分析吸烟对精子质膜完整性的影响并探讨其临床意义。方法:收集202例男性精液标本,其中132例为本院就诊男性不育患者,分为大烟量组(n=68)与小烟量组(n=64),正常生育男性为正常对照组(n=70)。通过计算机辅助精液分析系统进行精液常规分析。精液标本经PBS洗涤处理后用荧光染料SYBR-14/碘化丙锭(PI)双染后上流式细胞仪分析,用SYBR-14+/PI-(绿)表示质膜完整精子(PMI);SYBR-14-/PI+(红)表示质膜破损而坏死的精子;SYBR-14+/PI+(绿,红)表示正处于由活到死过度状态的精子。检测精子质膜完整性,并对精子质膜完整性与部分精液参数做相关性分析。结果:大烟量组与小烟量组质膜破损的坏死精子(SYBR-14-/PI+%)、过渡态的精子(SYBR-14+/PI-%)与正常对照组存在显著差异(P<0.01或0.05)。大烟量组SYBR-14+/PI-%明显低于小烟量组,SYBR-14-/PI+%明显高于小烟量组。所有202例标本SYBR14+/PI-%与精子活动率呈显著正相关(r=0.938,P=0.000)。结论:应用流式细胞术SYBR-14/PI双染法能快速、准确地检测精子质膜完整性。吸烟会导致精子质膜完整性下降,引起精子活力下降,可能是导致男性不育的重要原因之一。     

Effects of bovine sperm cryopreservation using different freezing techniques and cryoprotective agents on plasma, acrosomal and mitochondrial membranes

The success of semen cryopreservation is influenced by several factors, such as freezing curves and cryoprotectants. These two factors are of special interest once they may lead to many important physical-chemical changes resulting in different degrees of damage in spermatozoa structure. This experiment was designed to compare the effect of bull semen cryopreservation using two freezing techniques: conventional (CT--cooling rate of -0.55 °C min(-1) and freezing rate of -19.1 °C min(-1) and automated (AT--cooling rate of -0.23 °C min(-1) and freezing rate of -15 °C min(-1)), performed with different curves, and with three cryoprotectants (glycerol, ethylene glycol and dimethyl formamide) on bovine sperm motility and integrity of plasma, acrosomal and mitochondrial membranes. These variables were simultaneously evaluated using the fluorescence probes propidium iodide, fluorescein-conjugated Pisum sativum agglutinin and MitoTracker Green FM. The effects of freezing techniques, as well as of different cryoprotectants were analysed by the analysis of variance. The means were compared by Fisher's test. There were no significant differences between freezing techniques (P > 0.05). Glycerol showed higher percentages of motility, vigour and integrity of plasma, acrosomal and mitochondrial membranes than other two cryoprotectants (P < 0.05). Ethylene glycol preserved higher motility and integrity of plasma and mitochondrial membranes than dimethyl formamide (P < 0.05). Sperm motility with glycerol was 30.67 ± 1.41% and 30.50 ± 1.06%, with ethylene glycol was 21.17 ± 1.66% and 21.67 ± 1.13% and with dimethyl formamide was 8.33 ± 0.65% and 9.17 ± 0.72% to CT and AT curves, respectively. The percentage of spermatozoa with simultaneously intact plasma membrane, intact acrosome and mitochondrial function (IPIAH) was 14.82 ± 1.49% (CT) and 15.83 ± 1.26% (AT) to glycerol, 9.20 ± 1.31% (CT) and 9.92 ± 1.29% (AT) to ethylene glycol 4.65 ± 0.93% (CT) and 5.17 ± 0.87% (AT) to dimethyl formamide. Glycerol provided the best results, although nearly 85% of spermatozoa showed some degree of injury in their membranes, suggesting that further studies are required to improve the results of cryopreservation of bovine semen.     

添加黄芪注射液对人精子线粒体功能的影响

目的 观察添加黄芪注射液对人精子线粒体功能的影响。方法 弱精子症精子与黄芪注射液共孵化,用若丹明(Rh123)和碘化吡啶(PI)染色并用流式细胞技术检测人精子线粒体功能。结果 添加黄芪组线粒体功能良好的活精子数(9.831.65)明显高于对照组(6.500.41),与对照组比较差异有显著性(P<0.05)。添加黄芪组(42.493.73)的线粒体荧光值也明显高于对照组(31.074.49),与空白对照组比较差异有显著性(P<0.05)。结论 体外添加黄芪注射液能增强弱精子症精子线粒体的活性,为黄芪作为体外添加剂提高精子活力提供理论基础。     

Controlled cooling during semen cryopreservation does not induce capacitation of spermatozoa from two portions of the boar ejaculate

Cryopreservation imposes dramatic changes in boar sperm survivability but it is as yet unclear which part of the process affects the spermatozoa the most. The present study monitored, along the entire process of cryopreservation, the stability (PMS) of the architecture of the lipid plasma membrane and its integrity (PMI), as well as the kinetics of the processed spermatozoa using two portions from the boar ejaculate (P1 = the first 10 mL of the sperm-rich fraction, SRF; P2 = the rest of the ejaculate), frozen in a recently developed package, the MiniFlatPack (MFPs, 0.5 x 10(9) sperm/dose). Evaluation was made at four specific stages, viz. S1 = after collection (suspended in Beltsville thawing solution, BTS); S2 = at 15 degrees C (suspended in lactose-egg yolk, LEY); S3 = at 5 degrees C (suspended in LEY plus glycerol); and S4 = post-thaw. Both sperm kinetics (using computer-assisted sperm analysis, CASA) and PMS [i.e. the degree of lipid disorder and of the exteriorization of phosphatidylserine (PS) in the plasma membrane, measured by flow cytometry using Merocyanine-540 (M-540), and Annexin-V (AV) respectively], as well as plasma membrane integrity [PMI, i.e. the degree of membrane damage, measured using Yo-Pro-1 or propidium iodide (PI)] were assessed after incubation in BTS at 38 degrees C. Moreover, spermatozoa were challenged by incubation in modified Brackett-Oliphant medium (mBO+) with 37 mm of bicarbonate at 38 degrees C for 30 min, and their PMS and PMI further explored. Total sperm motility was significantly higher in P1 than in P2 along the entire process (S1-S4; p < 0.01), decreasing significantly at S4 for both fractions (p < 0.0001). The proportion of spermatozoa showing linear motility (LinM) was similar between ejaculate portions (P1 and P2), with a significant increase post-thaw (S4; p < 0.0001). During cooling (S1-S3) but not post-thaw (S4), lateral head displacement (LHD) differed between portions and changed along the stages (p < 0.01). Sperm velocity differed between portions in S1 (p < 0.01), but remained similar, independently of the portion, thereafter (S2-S4). Both PMS and the total number of live spermatozoa remained similar between S1 and S3 while incubated in BTS for both ejaculate portions. Sperm mortality increased post-thaw (S4) in both portions but the degree of lipid disorder remained low in the live cells (1.28% for P1; 1.55% for P2). Exposure to mBO+, on the other hand, significantly increased membrane lipid disorder along cooling (S1-S3; p < 0.0001), increasing the percentages of dead spermatozoa, especially post-thaw (around 70%, both portions). PS-exteriorization (AV) was not evident along the cryopreservation process in control (BTS) samples and exposure to mBO+ only induced minor variations. The data showed that kinetics, PMS and PMI of boar spermatozoa suspended in BTS (S1), LEY (S2) or LEY plus glycerol (S3) were maintained during controlled cooling but were altered by thawing, showing more characteristics of cell injury than of sperm capacitation. The spermatozoa were able to capacitate but the bicarbonate challenge destabilized the plasma membrane during initial cooling and accelerated membrane changes post-thaw. We conclude that capacitation of boar spermatozoa does not occur during controlled cooling.     

结核杆菌体外对人精子运动功能的影响

目的　研究结核杆菌体外对人精子运动功能的影响。方法　将从临床泌尿生殖系感染病人中分离培养出的结核杆菌制成活菌悬液 ,然后将其在体外与 10例健康成年男性手淫获得并经上游优化处理的精子按菌 /精比 5 0∶1一起孵化 0、2和 4小时后 ,采用CASA检测人精子活率、运动参数 (前向运动百分率、直线速度VCL、曲线速度VSL、平均路径速度VAP)和精子形态及凝集情况。结果　结核杆菌与精子比例为 5 0∶1体外孵化 0、2和 4小时后 ,人精子活率和运动参数无明显降低 (P >0 .0 5 )。结论　结核杆菌与精子的比例为 5 0∶1体外孵化 4小时内 ,对人精子的运动功能无显著影响     

Detecting subtle changes in sperm membranes in veterinary andrology

Thanks to the increasing use of flow cytometry in research in veterinary spermatology, many new membrane integrity assays have been developed over the past decade. These assays are important because of their superior ability to forecast fertility when compared with other tests, such as sperm motility. This major component of the sperm quality assessment has generated new investigations with the aim of developing tests that can detect membrane damage in a very early state. Using phospholipid transposition tests, early changes in membrane permeability and fluidity can be assessed in a large number of spermatozoa using fluorescent probes in combination with flow cytometry.     

精子质膜功能检测的研究进展

精子质膜上含有丰富的多聚不饱和脂肪酸及多种蛋白成分,精子质膜与精子获能、顶体反应及精卵融合密切相关。检测精子质膜功能,可预见精子的受精能力。评估精子质膜功能的方法主要有:精子尾部低渗肿胀试验、伊红Y水试验、精子膜脂过氧化反应的测定、精浆超氧化物歧化酶的测定、流式细胞术等。精子质膜功能检测可应用于检测精液质量、筛选精液离心法、评估性别分选精子质量和生育力、改进低温保存方法、优化卵细胞胞质内单精子注射(ICSI)技术。本篇综述将简要介绍精子质膜功能检测的原理、方法、步骤,并对其研究现状及应用作一概述。     

精子染色质结构分析的临床意义

目的探讨女方不明原因流产和不明原因不孕与其配偶精子染色质性状之间的关系;方法通过精子染色质结构分析(SCSA)对22例不明原因流产和36例不明原因不孕妇女配偶精子,以及20例正常对照者精子进行DNA完整性检测,并比较它们之间的差异;结果对照组与不明原因流产组以及不孕组之间精子DNA完整性存在差异,且差异具有统计学意义(P<0．05),而不明原因流产组与不孕组之间精子DNA完整性比较无明显差异(P>0．05);结论精子染色质结构分析有可能作为不明原因流产和不孕的辅助诊断方法在临床应用。