Testicular spermatozoon is superior to ejaculated spermatozoon for intracytoplasmic sperm injection to achieve pregnancy in infertile males with high sperm DNA damage

The purpose of this study was to compare the clinical outcome of testicular spermatozoon versus ejaculated spermatozoon in the treatment of infertile males with high sperm DNA damage, referred as sperm DNA fragmentation index (DFI), that attending intracytoplasmic sperm injection (ICSI) programme in terms of clinical pregnancy, births delivered as the primary and pregnancy loss and embryo fertilisation as the secondary outcome. A total of 102 males fulfilling the inclusion criteria were enrolled in the present study. Of the 102 males, 61 infertile males underwent testicular spermatozoon combined with ICSI while the remaining 41 males applied ejaculated spermatozoa in their first ICSI cycles, and the data of them were collected and analysed. In a 18‐month follow‐up, testicular spermatozoon achieved higher pregnancy rate and deliver rate than those in ejaculated sperm group (pregnancy rate, 36% vs. 14.6%, p = 0.017; deliver rate, 38.5% vs. 9.8%, p = 0.001). Nevertheless, there were no significant differences in the number of oocytes aspirated and number of embryos transferred between the two groups. Additionally, the fertilisation rate in the testicular sperm study cohort (70.4%) was also similar to that in the ejaculated sperm group (75.0%). Based on the current data, we conclude that testicular spermatozoon is the prior option in the treatment of infertile males with high sperm DFI in ICSI programme. More high‐quality studies with larger samples size are needed in the future due to the relative small size and the nonrandomized design of the present study.     

精子DNA碎片指数和精子畸形率对ICSI临床结局的影响

目的:通过评估ICSI治疗前的精子畸形率(SMR)和精子DNA碎片指数(DFI),探讨精子DFI和SMR对卵胞浆内单精子注射(ICSI)助孕结局的影响。方法:共入组79对因少弱精子症实施第一周期ICSI治疗的不孕夫妇,在进入治疗周期前3⁶个月,评价精子浓度、前向运动精子百分率、SMR及DFI。主要观察SMR和DFI与ICSI结局参数的关系。结果:79例少弱精子症患者DFI正常51例,异常28例,异常组的DFI值明显升高(14.18%vs 41.47%);巧合的是,SMR正常组同样为51例,异常组28例,异常组的SMR值亦明显升高(87.88%vs98.46%)。按DFI正常(DFI≤25%)与异常(DFI>25%)分组,或按SMR正常(≤96%)与异常(>96%)分组,组间的双方年龄、女方BMI、获卵数、移植胚胎数等基本情况差异无统计学意义。DFI正常和异常组间,SMR正常和异常组间的受精卵子数、可移植胚胎数、早期流产率无显著差异;异常组生化妊娠率(43.5%vs 61.5%)和临床妊娠率(39.1%vs 56.4%)降低,但差异无统计学意义(P=0.19及0.10)。精子DFI与SMR呈显著正相关(r=0.231,P<0.05)。结论:精子DFI增高(>25%),与按严格标准检测的SMR增高(>96%)男性行ICSI治疗,生化妊娠率和临床妊娠率降低,但与正常者比较未发现有统计学差异,可能与样本量小有关,有必要深入研究。     

Effect of male body mass index on clinical outcomes following assisted reproductive technology: a meta‐analysis

Overweight and obese males might exhibit a great risk of infertility. However, according to the current studies, the association between elevated male body mass index (BMI) and the clinical adverse results after assisted reproductive technology (ART) remains controversial. Hence, we conducted a meta‐analysis to evaluate the effects of raised male BMI on clinical outcomes following ART. PubMed, EMBASE and three Chinese databases were used to identify relevant studies. The primary outcome was clinical pregnancy rate. Secondary outcomes included live birth rate and sperm parameters. A total of 5262 male participants from 10 cohort studies were subjected to meta‐analysis. Results indicated that overweight or obese had no significant impact on clinical pregnancy rate [in vitro fertilisation (IVF): odds ratio (OR), 0.73; 95% confidence interval (CI), 0.39–1.39; intracytoplasmic sperm injection (ICSI): OR, 1.03; 95% CI, 0.92–1.15], live birth rate (IVF: OR, 0.91; 95% CI, 0.78–1.06; ICSI: OR, 1.00; 95% CI, 0.50–1.99) and sperm concentration (SMD, ?0.28; 95% CI, ?0.65 to 0.08) compared with normal weight following IVF/ICSI treatments. Exclusion of any single study and almost all the sensitivity analyses showed that our results were reliable. At present, the role of male BMI in the process of ART is only partly understood and should be further investigated.     

Sperm DNA fragmentation measured by Halosperm does not impact on embryo quality and ongoing pregnancy rates in IVF/ICSI treatments

Sperm DNA fragmentation (SDF) has been proposed to be one of the main markers regarding male infertility. A prospective study was performed to assess primarily whether sperm DNA damage has any impact on embryological data and secondarily on pregnancy rates. This prospective study evaluated the sperm DNA damage in fresh ejaculated sperm samples from couples undergoing IVF/ICSI treatments, using the improved SCD method, known as Halosperm^®. The results were evaluated by performing statistical analysis with the statistical package of SPSS v17. A total of 156 fresh semen samples derived from 156 couples undergoing 156 IVF/ICSI cycles. From the 156 couples, 139 finally reached the embryo transfer (ET) procedure. Overall, SDF did not correlate with embryological data, while ongoing pregnancy rate/ET was 21.6%. SDF only correlated with sperm characteristics. After the categorisation of SDF (≤35% and >35%), according to the specific references of the method used, embryological data were comparable as also ongoing pregnancy rates. Using the SCD method, sperm DNA damage is associated neither with embryological data nor to pregnancy rates. However, we should not rule out the fact that extremely high DNA damages are associated with total pregnancy failure.     

不同来源精子对ICSI治疗后妊娠结局的影响

目的分析不同精子来源和数量对单精子卵胞浆内注射术(ICSI)妊娠结局的影响。方法2000年1月～2005年1月在本中心进行ICSI治疗而妊娠的354例,据精子来源与数量分为精液正常组(47例)、少弱精子组(194例)与手术取精子组(113例)。比较3组的临床妊娠率、流产率、分娩率、妊娠并发症、分娩孕周、新生儿出生体重、畸形、围产期死亡率等组间差异。结果3组患者的年龄、不孕年限、临床妊娠率(41.3%、39.7%、40.4%)、流产率(14.9%、18.6%、15.9%)、分娩率(78.7%、75.8%、79.6%)、双胎率(34.4%、35.4%、37.8%)、妊高症发生率(10.8%、10.2%、11.1%)、前置胎盘发生率(5.4%、6.8%、6.7%)、早产率(27.0%、27.9%、27.8%)、分娩孕周(36.4±5.4)w、(37.6±2.3)w、(37.2±2.8)w、胎儿出生体重(2765.4±624.9)g、(2749.0±523.5)g、(2763.5±684.4)g、先天性畸形发生率(4.5%、5.0%、4.0%)、围产期死亡率(40‰、45‰、48‰),差异均无显著性(P>0.05)。结论不同精子来源与数量不影响ICSI治疗后的妊娠和围产儿结局。     

Influence of bacteria and leukocytes on the outcome of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI)

Summary. The influence of bacteria and/or leukocytes on the outcome of IVF or ICSI is influenced by three factors which have little in common with in vivo conditions: 1) The process of ejaculate preparation (swim-up, Percoll) with antibiotic buffered media; 2) The small amount of inseminated spermatozoa (100000 per culture); 3) The short cultivation time. From the very beginning, these factors limit whatever the influence of bacteria and leukocytes on fertilization and embryonic development in vivo may be. Despite the contradictory results published so far, the influence of bacteria and/or leukocytes on the functional integrity of spermatozoa during the process of IVF or ICSI can be ignored. Furthermore, during IVF or ICSI the spermatozoon does not act as a vector for the transportation of bacteria into the ooplasm.     

Fertilisation and pregnancy outcome after ICSI in globozoospermic patients without assisted oocyte activation

The successful outcome of intracytoplasmic sperm injection (ICSI) with globozoospermic sperm and non-activated oocytes is reported. Three couples underwent ICSI treatment and two of the patients were siblings. Forty-four non-activated oocytes were injected, 26 oocytes fertilised normally and 17 good quality embryos were obtained. Six embryo transfers were carried out, three with fresh embryos and three with frozen-thawed embryos. Three pregnancies resulted from the fresh embryo transfers and additionally two pregnancies were obtained after the transfer of frozen-thawed embryos. Two healthy babies were born. One twin pregnancy is ongoing. Our case reports demonstrate that in some ICSI attempts undertaken with globozoospermic sperm cells from two of our patients, high fertilisation rates, pregnancies and live births can be achieved, without artificially activated oocytes. Our data also suggest that in some cases, round-headed spermatozoa lack the capacity to activate the oocyte. Therefore, it cannot be excluded that artificial oocyte activation could be of help in globozoospermic patients with complete fertilisation failure.     

不同受精方式对选择性囊胚培养结局的影响

目的:探讨不同受精方式对选择性囊胚培养结局的影响。方法:回顾性分析2009年1月至2012年12月在我中心接受囊胚培养的IVF组1 153个周期和ICSI组205个周期的囊胚培养结局。结果:IVF组和ICSI组分别有14 748和2 655枚胚胎进行序贯囊胚培养,两组均无因无囊胚可移植而取消周期。IVF组共形成7 871枚囊胚,ICSI组形成1 210枚囊胚;IVF组的优胚率、囊胚形成率和胚胎利用率均显著高于ICSI组(64.77%vs 58.72%,53.37%vs 45.57%,60.06%vs 52.17%,P均0.05);但IVF组与ICSI组种植率(48.94%vs51.43%)、临床妊娠率(49.03%vs 52.02%)和流产率(11.69%vs 15.56)均无显著差异(P均0.05)。结论:在同样的选择性囊胚培养纳入标准下,IVF相比于ICSI行囊胚培养的囊胚形成率和胚胎利用率更高,因无囊胚移植取消周期的风险相对更小。针对不同的受精方式可能需要制定相应的选择性囊胚培养纳入标准。     

完全受精失败周期应用早期补救性卵细胞胞质内单精子注射术的安全性评估

目的:评估完全受精失败周期应用早期补救性单精子卵细胞胞质内注射(ICSI)术的安全性。方法:105例早期补救性ICSI周期,随访临床结局。结果:早期补救性ICSI周期出生婴儿64名。出生婴儿的性别比无偏差,出生体重正常,无严重出生缺陷。结论:早期补救性ICSI没有增加临床应用的安全性风险,临床应用前景十分广阔。     

Factors affecting successful outcome from ICSI in men following previous vasectomy

There are conflicting reports as to whether the interval between vasectomy and surgical sperm retrieval (SSR) for intra-cytoplasmic sperm injection (ISCI) is related to clinical pregnancy (CPR), and live birth (LBR), rates. This study aimed to evaluate factors that may influence the outcome of ICSI in males with secondary azoospermia due to previous vasectomy. We analysed the medical records of 198 azoospermic males following vasectomy who underwent percutaneous epididymal sperm aspiration (PESA) and/or testicular sperm extraction (TeSE), between 1997 and 2005 by a single urologist, and whose sperm was subsequently frozen for use in an IVF treatment programme on their partner’s behalf. Hundred and forty-four (73%) males had a positive PESA, and the remaining 54 (27%) had a positive TeSE. Forty-four percent of males with no clinical evidence of epididymal distension still had epididymal sperm retrieved successfully. Hundred and twenty-eight patients proceeded with ICSI, and a total of 237 cycles were performed. The CPR and LBR overall were 29 and 27%, respectively. Using logistic regression there was no association between time since vasectomy and CPR (P = 0.17) or LBR (P = 0.31). A history of an attempted reversal of vasectomy did not negatively affect retrieval rates or CPR and LBR. The success of SSR and the outcome of ICSI, using frozen sperm, are independent of male age and time since vasectomy. Epididymal sperm may be retrieved in over 40% of men in whom there is no clinical evidence of epididymal distension.     

梗阻性无精子症患者睾丸抽吸精子行ICSI后的胚胎结局

目的总结非手术精子抽吸(NSA)联合ICSI对梗阻性无精子症患者的治疗意义,并探讨精子来源等对ICSI后胚胎结局的影响。方法回顾性分析642个ICSI周期,比较了睾丸抽吸精子和精液精子行ICSI后的受精率、优质胚胎率、临床妊娠率、种植率及胚胎停育率等。结果NSA睾丸精子组和精液精子组相比较,其2PN受精率、优质胚胎率、临床妊娠率、种植率及胚胎停育率之间均没有显著性差异(71．4％vs 73．2％, 73．1％vs 70．1％,34．2％vs 32．6％,22．3％vs 19．8％,21．3％vs 25．0％,P>0．05)。结论非手术抽吸睾丸精子联合ICSI是治疗梗阻性无精子症的一种有效方法。     

Comparison of conventional in vitro fertilisation and intracytoplasmic sperm injection outcomes in patients with moderate oligoasthenozoospermia

The method of choice for assisted reproductive technology treatment in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) is usually based on the evaluation of male infertility factors. Decisions for couples with moderate oligoasthenozoospermia (OA) are often empirical because uniform treatment criteria are lacking. This study aimed to evaluate the effect of patients with moderate OA treated with conventional IVF and ICSI. A total of 199 couples with moderate OA undergoing their first IVF/ICSI cycle were included in the study. The patients were divided into two groups according to the type of insemination: conventional IVF group (n = 97) and ICSI group (n = 102). All patients were randomised to be inseminated either by conventional IVF or ICSI. The fertilisation rate, embryo quality, implantation rate and clinical pregnancy rate were examined. No differences in the fertilisation, implantation and pregnancy rates were observed between conventional IVF and ICSI groups (P > 0.05). However, the number of good‐quality embryos was significantly higher in the ICSI group than in the IVF group (P < 0.05). Couples with moderate OA did not influence on the overall clinical outcomes between IVF and ICSI treatments, and a negative influence by ICSI on blastocyst development was not confirmed.     

The effect of sperm DNA fragmentation on intracytoplasmic sperm injection outcome

Our study objective was to assess the effect of various sperm DNA fragmentation levels on clinical intracytoplasmic sperm injection outcome. This retrospective study included 392 patients who underwent ICSI and performed sperm DNA fragmentation testing before the procedure. Based on sperm DNA fragmentation cut-off values, the patients were differentiated into 3 groups as <20%, 20%–30% and >30%. According to the female status, patients were differentiated into favourable group (n = 259) with female age <35 years and anti-Mullerian hormone level ≥7.1 pmol/L; and unfavourable group (n = 133) with female age ≥35 years and anti-Mullerian hormone level ≤7.1 pmol/L. The patient's medical records were reviewed, and patient's demographic, laboratory data including semen analysis, sperm DNA fragmentation determined by means of sperm chromatin dispersion, hormonal profile and data regarding intracytoplasmic sperm injection cycle were collected. This cohort reported that the clinical reproductive outcomes of intracytoplasmic sperm injection showed no statistical significance with increase sperm DNA fragmentation levels. In sperm DNA fragmentation above 30%, favourable females had significantly higher clinical pregnancy rate and live birth rate than unfavourable females, while fertilisation rate and miscarriage rate showed no significance between the subgroups. High sperm DNA fragmentation is linked to poor semen parameters.     

An additional marker for sperm DNA quality evaluation in spermatozoa of male partners of couples undergoing assisted reproduction technique (IVF/ICSI): Protamine ratio

The aim of this study was to evaluate the relationship between the protamine ratio (P1/P2), DNA fragmentation of spermatozoa and protamine deficiency. Patients were grouped into fertile (G1; n = 151) and sub‐fertile (G2; n = 121). DNA fragmentation in spermatozoa was analysed by a TUNEL assay (terminal deoxynucleotidyl transferase‐mediated deoxyuridine triphosphate nick‐end labelling), and the protamination was determined by CMA3 staining, while Western blot was used to measure protamine P1 and P2. While sperm DNA fragmentation (SDF) and protamine ratio were significantly elevated in G2 compared with G1 (12.31 ± 7.01% vs. 17.5 ± 9.5%; p = .001) and (0.91 ± 0.43 vs. 0.75 ± 0.42; p = .003); respectively, the CMA3 positive showed no difference at all between G1 and G2. In G1, the CMA3 positive correlated negatively with the P1/P2 ratio and SDF (r = ?.586, r = ?.297; p = .001 respectively). In contrast, the protamine ratio correlated positively with SDF (r = .356; p = .001). In G2, no correlation was observed between CMA3 positive, SDF and the P1/P2 ratio but the P1/P2 ratio showed a positive correlation with SDF (r = .479; p = .001). In conclusion, the spermatozoa DNA deterioration was closely associated with abnormal protamination but showed an association with the protamine ratio, more than with CMA3 positive. Therefore, for the evaluation of DNA damage in spermatozoa, the P1/P2 ratio might act as an additional biomarker.     

卵胞浆内单精子注射和常规体外受精-胚胎移植的胚胎形态及临床结果比较

目的 :比较卵胞浆内单精子注射 (ICSI)和常规体外受精 -胚胎移植 (IVF-ET)两种助孕技术处理后的胚胎形态和临床结果。方法 :90对夫妇进行 93个 ICSI/ IVF治疗周期 (ICSI 45周期 ,IVF 48周期 ) ,比较两组周期平均获卵数 ,正常受精数 ,A、B级胚胎数和生化妊娠率 ,临床妊娠率。结果 :ICSI组周期平均获卵数及正常受精数 (分别为 1 1 .2 5 ,6 .6 0 )与 IVF组 (分别为 1 0 .6 7,6 .90 )相类似 (P>0 .0 5 ) ;周期 A、B级胚胎数 ICSI组3.31 ,IVF组 3.1 9,两组比较无显著差异 (P>0 .0 5 )。周期临床妊娠率 ICSI组 42 % ,IVF组 44 % ,两组比较无显著差异 (P>0 .0 5 )。周期“抱婴回家”率 ICSI组 36 % ,IVF组 31 % ,两组比较无显著差异 (P>0 .0 5 )。结论 :ICSI技术治疗严重男性因素引起的不育症可取得与常规 IVF治疗主要由女方不育因素引起的不育症相似的临床效果     

卵细胞胞质内单精子注射在前次常规体外受精失败病例中的应用

目的:探讨卵细胞胞质内单精子注射(ICSI)是否可以改善由于受精障碍导致不孕病例的妊娠结局。方法:回顾性分析2002年1月~2004年12月由于第1周期的常规体外受精(IVF)治疗中卵子完全不受精或受精率≤20%,而在随后的治疗周期中采取ICSI方法受精的20个病例(共22个周期,研究组)的ICSI治疗结局。对照组为同期因男方少弱精子进行第1次ICSI治疗的连续100个周期。结果:与前次IVF相比,经ICSI治疗后受精率从5.4%显著提高至76.9%(χ2=264.66,P<0.001),但研究组中低受精患者经ICSI治疗后的受精率67.9%,明显低于对照组的77.5%(χ2=5.18,P<0.05)和研究组中完全不受精患者的84.2%(χ2=8.97,P<0.05)。研究组中完全不受精患者的妊娠率和胚胎种植率分别为57.1%和40.5%,均高于对照组的35.0%和18.9%,但仅胚胎种植率的差异有显著性意义(χ2=8.80,P<0.01)。结论:ICSI可以有效地克服受精障碍,提高受精率,并且可明显改善妊娠结局。     

Outcomes in couples undergoing ICSI: comparison between fresh and frozen-thawed surgically retrieved spermatozoa

We retrospectively evaluated the impact of cryopreservation on spermatozoa obtained from patients with azoospermia and used for intracytoplasmic sperm injection (ICSI). Frozen-thawed epididymal spermatozoa (FTEPS) was used in 34 couples, whereas frozen-thawed testicular spermatozoa (FTTS) was used in 50 couples for ICSI during assisted conception, and these results were compared with results using fresh spermatozoa for ICSI in the same individuals. The fertilization rate (FR) was significantly lower for FTTS (65.8%) but not for FTEPS (73.1%) compared with the FR using fresh spermatozoa (72.3% and 73.2% respectively). In contrast, neither the implantation nor the pregnancy rate was altered when FTEPS or FTTS was used. In conclusion, our results indicate that surgically retrieved spermatozoa can be efficiently used for ICSI after freezing and thawing without compromising the outcome.     

Clinical management of infertile men with nonobstructive azoospermia

The clinical management of men with nonobstructive azoospermia (NOA) seeking fertility has been a challenge for andrologists, urologists, and reproductive medicine specialists alike. This review presents a personal perspective on the clinical management of NOA, including the lessons learned over 15 years dealing with this male infertility condition. A five-consecutive-step algorithm is proposed to manage such patients. First, a differential diagnosis of azoospermia is made to confirm/establish that NOA is due to spermatogenic failure. Second, genetic testing is carried out not only to detect the males in whom NOA is caused by microdeletions of the long arm of the Y chromosome, but also to counsel the affected patients about their chances of having success in sperm retrieval. Third, it is determined whether any intervention prior to a surgical retrieval attempt may be used to increase sperm production. Fourth, the most effective and efficient retrieval method is selected to search for testicular sperm. Lastly, state-of-art laboratory techniques are applied in the handling of retrieved gametes and cultivating the embryos resulting from sperm injections. A coordinated multidisciplinary effort is key to offer the best possible chance of achieving a biological offspring to males with NOA.     

A report of pregnancy following ICSI in one of two sisters with familiar primary ciliary dyskinesia

Primary ciliary dyskinesia (PCD) is a disorder of structure and function of motor ciliary and dyskinetic activity of ciliary in the fallopian tubes of affected women and could lead to infertility in some cases. In vitro fertilisation (IVF) is a choice of treatment in infertile women with PCD, which could conquer the tubal dysfunction. In this case study, we report a PCD affected woman with infertility who was treated by IVF and pregnancy was achieved but it failed due to the spontaneous abortion. We also performed whole-exome sequencing for this case and her PCD-affected sister, which did not reveal any genetic abnormality related to the PCD or infertility.     

常规体外受精失败后6h补行卵胞浆内单精子注射及其严格指征的临床意义

目的探讨为进一步严格控制卵胞浆内单精子注射（ICSI）指征,在常规体外受精（IVF）失败后6h补行ICSI的方法对临床结果的影响。方法回顾分析本中心2006年1月至2007年11月常规IVF或ICSI的方法进行的治疗周期（A组）与2007年12月至2008年11月修改ICSI指征后,使用常规IVF及受精失败后6h补行ICSI的方法进行的治疗周期（B组）比较两个时期的总周期的临床妊娠率和ICSI所占比例。结果A组799周期,临床妊娠率为35．29％,其中行ICSI369例,占总周期46．18％;B组704周期,临床妊娠率为42．19％,其中行ICSI190例,占总周期26．99％。两组的总临床妊娠率有显著性差异,行ICSI的比例也有显著性差异（P〈0．05）。结论使用常规IVF受精失败后6h补行ICSI的方法,修改ICSI指征,ICSI比例降低,对本中心的临床妊娠率无影响。