两种精子处理方法的卵细胞胞质内单精子显微注射结局的比较

目的:评价密度梯度离心和改良上游法两种处理活动精子的分离方法在卵细胞胞质内单精子显微注射(ICSI)中的效果,从而指导临床应用。方法:选取2004年10月~2005年4月在本中心完成的42例患者42个周期为研究对象,前瞻性比较了两种精子分离方法的受精率、卵裂率、优质胚胎率、临床妊娠率、精子畸形率、精子回吸收率等。结果:两种方法分离精子所获得的ICSI胚胎移植(ICSI-ET)中,受精率、卵裂率、优胚率、临床妊娠率均无明显差异,但是改良上游法所获得的精子畸形率明显高于密度梯度离心法(P<0.01);重度少精子症密度梯度离心法回吸收精子优于改良上游法(P<0.01)。结论:在辅助生育技术ICSI-ET中,密度梯度离心法分离活动精子临床妊娠结局与改良上游法无明显差异;除对重度少精子症者外,均可以采用改良上游法。     

不同处理方法优选精子效果的比较

目的 比较上游法、三种密度梯度离心法和洗涤离心法优选精子的效果. 方法 门诊收集浓度与活力正常的精液标本44份,每份精液各取0.5 ml×5,分别采用上游法、密度梯度离心法和洗涤离心法进行处理,比较各组的精子浓度、活力、畸形率、精子畸形指数（SDI）和DNA碎片指数（DFI）,再按试剂来源将密度梯度离心法分为Sperm Grad梯度离心组（A组）、Pure Sperm梯度离心组（B组）和Pure Ception梯度离心组（C组）,进行上述参数比较. 结果与上游法和梯度离心法相比,洗涤离心法精子浓度、DFI值、畸形率、SDI值及头部、中段、尾部畸形率均显著增高（P＜0.05）,活力显著降低（P＜0.05）;与梯度离心法相比,上游法精子浓度、DFI值均显著降低（P＜0.05）,活力显著升高（P＜0.05）,两组间总畸形率、SDI值及头部、中段、尾部畸形率均无显著性差异（P＞0.05）.A、B、C组间精子浓度、活力、畸形率、SDI及DFI等参数均无显著性差异（P＞0.05）,但C组中段畸形率显著高于A组和B组（P＜0.05）,B组尾部畸形率显著高于A组和C组（P＜0.05）; 结论对于精子浓度和活力正常的精液标本,上游法精子活力最高、畸形率和DFI值最低,且精子浓度能够满足临床需要,是较为理想的精子优选技术;三种不同梯度离心法优选精子效果无显著性差异,适用于严重少精子症精液标本的处理.     

两种精子优选方法效果比较

目的　探讨精子经王氏管和密度梯度离心法处理后精子参数的变化 ,比较两者在宫内人工授精 (IUI)的临床效果。　方法　选择不育男性精液 15 7份 ,采用两种方法进行配对处理 ,比较两种分离方法前后精子活力、正常形态率、精子顶体形态和精子染色质等变化 ;分离后的精子用于IUI的临床妊娠率。　结果　两种方法处理后的精子活力 (a +b级 )、正常精子形态率、精子顶体完整率、正常染色质率与处理前比较有显著差异 (P <0 .0 1) ;但两种方法分离的精子用于IUI的临床妊娠率无显著差异 (P >0 .0 5 )。　结论　王氏管法和密度梯度离心法处理精子后均获得质量较好的精子 ,但用于IUI后临床妊娠率无明显差异     

精子分离两种方法的比较和评价

目的 :探讨王氏管法和精子上游法在精子分离中的应用 ,比较两者的优缺点。　方法 :随机选择 12 7份精液 ,采用 2种方法进行配对处理 ,比较 2种方法分离前后精子活动率、形态、活力和精子顶体形态、精子染色质以及精子膜完整性等参数的变化。　结果 :2种方法处理后的精子活动率、精子活力以及精子形态有显著的差异 (P <0 0 1) ,王氏管法处理后精子顶体完整率、正常染色质率及精子膜完整率亦显示出显著的差异 (P <0 0 1)。　结论 :王氏管法和精子上游法在精子处理中均可获得较好的精子质量 ,但王氏管法则获得更优质的精子 ,且具有方便操作和实用的特点。     

三种精液处理方法所得精子的体外观察分析

精液处理是人工授精和试管婴儿过程中重要的一环,优化处理精液是为了提高精子的密度和活力,并去除精浆内的前列腺素、抗精子抗体、炎症细胞等不良成分。为寻求简便、高效、快速的精液处理方法,本研究对三种精液处理方法(Earle’s上游法、双层Percoll梯度离心法和单层Percoll高速离心法)进行对比研究,现报道如下:     

两种精液处理方法对精液参数及冷冻效果的比较分析

目的 比较两种精子处理方法的优劣.方法 55例精液标本随机分为直接离心组(30例)和加精子洗涤液离心组(25例),进行精液处理后并且冷冻,观察其复苏效果,所有样本均进行精液常规分析.结果 直接离心组精子密度由46.52×106/ml提高到86.50×106/ml,而前向运动精子(a+b)级百分率则由58.00%下降到44.33%;加精子洗涤液离心组精子密度由44.44×106/ml提高到85.96×106/ml,而前向运动精子(a+b)级百分率则由57.76%下降到46.48%,两组间均无显著性差异(P>0.05).对两组处理后的精液进行冷冻,复苏后得到其前向运动精子复苏率均在40%左右,两组间差异无统计学意义(P>0.05).结论 两种处理方法具有相近的浓缩效果和前向运动精子复苏率,直接离心法则更为简便且节省成本.     

不孕症患者宫腔内人工授精精子优选方法分析

目的:比较分析精子不同优选方法与宫腔内人工授精(IUI)成功率的相关性,以提高IUI妊娠率。方法:回顾性分析本院生殖中心门诊不孕症患者452例671个IUI周期,按精子优选方法分为3组:含5%人血清白蛋白(HSA)的Earle's液上游法、SpermR inse上游法、SupraSperm密度梯度离心法。比较各组患者的处理前后精子密度与活动率及临床妊娠结局。结果:3种精子优选方法处理后精子活动率及a+b级精子百分率显著增加(P<0.01)。含5%HSA的Earle's液上游法221个IUI周期,妊娠21个周期,周期临床妊娠率9.5%;SpermR inse上游法215个IUI周期,妊娠34个周期,周期临床妊娠率15.8%,与含5%HSA的Earle's液上游法比较有显著性差异(P<0.05);SupraSperm密度梯度离心法235个IUI周期,妊娠34个周期,周期临床妊娠率14.5%,与含5%HSA的Earle's液上游法比较无统计学差异(P>0.05)。结论:SpermR inse上游法的IUI临床妊娠率明显高于含5%HSA的Earle's液上游法,适用于各类不孕症患者。SupraSperm密度梯度离心法也具有较高的周期临床妊娠率,洗涤效果好,操作便捷高效,尤其适用于含炎症细胞、死精子和畸形精子等不良成分较多的精液。     

精液上游分离法对精子正常形态和核完整性的作用

精子分离制备技术的应用由来已久,上游分离法具有操作简便、成本低廉、副反应少等优点仍被一些辅助生殖技术实验室采用。该方法主要有助于富集前向运动精子及减少标本中的非精子细胞成分。本文对上游分离法处理前后的精液中正常形态精子和核完整精子的百分比进行了比较,以了解上游分离法对该两项指标的效果。     

冰箱冷冻法和程序降温仪冷冻法对人类精液冷冻复苏后精子参数的影响

<正>近些年,随着更多的精子库在国内得到卫生部的批准而成立,精液的冷冻技术也有了很大的提高,除了世界卫生组织《人类精液检查与处理实验室手册》(第5版)推荐的冷冻方法外[1],还衍生出了更多的其他精液冷冻方法[2-3]。程序降温仪由于能够精确控制液氮蒸气注入冷冻室的速率,降温速率精确,仍然为首选的冷冻方法。但由于人们对精子库认识的加深和理解,更多的捐精志愿者来到精子库进行精液的捐献,而仅靠程序降温仪已经不能满足     

磁性活性细胞分选技术在男性不育研究中的应用

磁性活性细胞分选法(MACS)根据细胞表面特异的标记物,在分子水平对目的细胞进行有效分选,具有简易、快速、灵活、特异性高的特点,在临床方面有着广泛的应用。MACS也为男性不育提供了一个新的研究平台,将MACS用于精液质量优化与生殖细胞分离是男性不育研究的一个新思路。本文简要介绍了MACS的基本原理,综述了MACS在精子优选、冷冻保存、精原干细胞及生精细胞分离等男性不育研究方面的应用现状和临床应用前景。     

Comparison of four methods for sperm preparation for IUI

We compared the pregnancy rates and sperm characteristics of semen prepared by the albumin, Percoll, Puresperm, and swim-up methods. Semen analysis was performed by a computer-assisted system (CASA). The overall pregnancy rate was 14% per patient and 11% per cycle. The albumin and Percoll methods had the highest pregnancy rates, up to 12% per cycle. We then compared each method with the albumin method. The swim-up technique yielded the highest percentage of motile sperm. The Percoll method yielded statistically significant changes in average path velocity and straight-line velocity, while the Puresperm method revealed the same statistical changes in total concentration and hyperactive motile sperm percentage. After adjusting for age, methods, and CASA estimates, only straight-line velocity was significantly correlated with pregnancy. Although the Percoll method was associated with the best pregnancy rate, this product has been withdrawn from the market because of toxic contamination. Puresperm had replaced it in our laboratory but showed an unsatisfactory pregnancy rate. The swim-up method is the best choice for IUI.     

四种精液检测方法的比较和临床评价

目的 对以色列计数板(IC)、Makler计数板(MC)、精子质量分析仪(SQA)和血细胞计数板(CC)应用于精液常规分析的结果进行比较，并作出评估。方法 用4四种方法对来自有正常生育力的健康男性的64份精液标本进行分析。结果 在计数精子密度方面，IC、MC和SQA的结果有较好的一致性，3者之间无统计学上显著性差异；而CC与IC和SQA之间存在统计学上显著性差异，其P值分别为0．002和0．005。在计数精子活率方面，CC与MC和SQA之间也有统计学上显著性差异，其P值分别为0和0．02。我们使用了IC和MC检测前向运动精子百分率，结果显示两者之间无统计学上显著性差异，两者之间有较好的一致性。结论 在精液常规分析中，IC、MC和SQA这3种方法的结果虽具有较好的一致性，但不如CC准确，在临床应用时，应注明使用何种方法进行测定；也可将IC、MC和SQA等方法与CC进行比较，计算出校正系数予以调整。     

Comparison of intracytoplastic sperm injection outcomes with fresh and frozen-thawed testicular sperm

Testicular sperm can be obtained from patients with non-obstructive azoospermia. However, there is not enough evidence concerning whether fresh or frozen-thawed sperm is better for successful pregnancy. We retrospectively compared initial treatment results of intracytoplasmic sperm injection using fresh and frozen-thawed testicular sperm in our institution. From August 1997 to May 2006, a total of 27 cases including 18 cases using fresh sperm (269 oocytes) and 9 cases using frozen sperm (97 oocytes) were evaluated. In the fresh and frozen sperm groups 33.4 and 32.9%, respectively, developed good quality embryo. There was no significant difference between the two groups in successful pregnancy and birth rates.     

Comparison between the quality and function of sperm after semen processing with two different methods

Aim: To compare the recovery rate of morphologically normal and chromatin condensed spermatozoa from native se-men samples using the SpermPrep~(TM) filtration columns and Percoll gradient centrifugation and to determine the influenceof the two processing techniques on fertilization and pregnancy rates in an IVF-ET program. Methods: Sixteen se-men samples obtained from patient's husband were included in this study. Each was divided into two aliquots. The firstaliquot was processed with SpermPrep~(TM) filtration columns and the second, Percoll gradient centrifugation. Smears weremade before and after semen processing with both methods for the evaluation of chromatin condensation (chromomycineCMA3) as well as morphology (strict criteria) of spermatozoa. One hundred and seventy oocytes were retrieved fromthe patients and the oocytes from each patient were subdivided into two sets : one set was inseminated using spermatozoaprocessed with SpermPrep~(TM) and the other inseminated after semen processing with Pe     

掌指骨骨折的手术治疗

目的：探讨合理、有效的掌指骨骨折的治疗方法，以期提高治疗效果。方法：本组295例，男228例，女67例；年龄14～65岁，平均29岁。采用钢丝固定（45例，A组）、微型接骨钢板固定（65例，B组）、单根克氏针内固定（97例，C组）及双根克氏针内固定（88例，D组）4种方法治疗，并对不同的治疗方法及效果进行评定。结果：术后平均随访18个月（6～72个月），按掌指关节伸屈正常范围标准进行评定，优100例，良165例，可17例，差13例。A组：优12例，良24例，可4例，差5例，优良率80．0％；B组：优20例，良37例，可5例，差3例，优良率87．6％；C组：优38例，良55例，可2例，差2例，优良率95．8％；D组：优30例，良49例，可6例，差3例，优良率89．7％。结论：钢丝法不应作为常规治疗和首选，仅适合于掌指骨横形骨折；微型钢板对多发性、移位骨折可取得良好效果；对于大多数手部骨折，克氏针固定是有效、可靠的手术方式。     

Comparison of four methods to evaluate sperm DNA integrity between mouse caput and cauda epididymidis

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.     

精液保存和优化处理方法对精子DNA完整性的影响

目的:探讨不同精液保存和处理方法对精子DNA完整性的影响。方法:筛选2015年1~12月就诊的100例精液正常志愿者,将每例志愿者的精液混匀后分别对其进行不同的保存和优化处理;保存方法包括直接冷冻法、试剂冷冻法、室温保存4 h和24 h等,优化处理方法包括简单洗涤法、直接上游法和非连续密度梯度离心法。保存和处理后均采用精子染色体扩散实验(SCD)分析其精子DNA碎片指数(DFI);优化处理各组继续培养24 h,培养后再分析其精子活动率和DFI。结果:精液保存条件两两组间分析显示,直接冷冻法、试剂冷冻法和室温保存4 h的精子DFI分别为(27.3±6.4)%、(26.9±6.1)%和(24.7±6.8)%,结果无显著性差异(P0.05),而室温保存24 h则显著增加精子DFI[(35.6±9.0)%](P0.05)。与简单洗涤法的精子DFI[(13.7±2.0)%]相比,直接上游法的精子DFI[(9.1±1.3)%]和非连续密度梯度离心法的精子DFI[(8.0±2.5)%]均显著降低(P0.05);再培养24 h后,非连续密度梯度离心法处理的精子DFI[(11.5±4.2)%]最低(P0.05)。结论:保存条件和优化处理方法对精子DNA完整性有影响,临床工作中需选取最合适的精液处理方法。     

Autostage sperm tracing system for semen evaluation

To overcome the limitation of the microscope field, the study proposed an autostage sperm tracing system (ASTS), which could trace a particular sperm for a long time and distance. The ASTS was constructed by assembling a commercial microscope, an image frame grabber, a personal computer, and a motorized stage. Its performance was tested by evaluating 6 semen samples and by comparing the evaluation with those of other semen evaluations. The ASTS broke through the limitation of the microscope field and traced a particular sperm as long as possible. It analyzed the sperm track and calculated the motility parameters, such as curvilinear velocity (Vcl), straight-line velocity (Vsl), and linearity (L(in)). The sperm quality was then evaluated in real time, and the user could decide to capture or abandon a particular sperm in the IVF The ASTS enables users to evaluate sperm progression for a long time and to have the global quality of a particular sperm in real time. Its open structure has the flexibility for micromanipulating a semen sample, and has the potential application associated with a modern IVF technique.