Effect of Vitamin D Supplementation on Intracytoplasmic Sperm Injection Outcomes: A Randomized Double-Blinded Placebo-Controlled Trial

OBJECTIVE: To assess the effect of vitamin D supplementation on enhancement of embryos implantation rates in intracytoplasmic sperm injection procedures. STUDY DESIGN: A randomized double-blinded controlled trial. SETTING: Assisted Reproduction Unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Egypt. DURATION: From December 1st 2017 to the end of October 2018. SUBJECTS and METHDS: Eighty infertile couples that underwent ICSI procedure were entrapped in this study. Patients were divided into two groups;Group I (Vitamin D group) = 40 patients (received vitamin D supplementation in the form of Vidrop oral drops 600 IU/day starting after ovum pickup) and Group II (placebo group) = 40 patients (received a placebo “normal saline oral drop”). RESULTS: There was a significantly increased embryo implantation rates (53.2% in vitamin D group versus 46.7% in placebo group), there was a mild statistically significant difference in the clinical pregnancy rates (45.9% in vitamin D group versus 39.4% in placebo group) with p-value < 0.05 and there was an improvement in ongoing pregnancy rate as there was a mild statistically significant difference in first trimester miscarriage rates (12.5% in vitamin D group versus 17.5% in placebo group) with p value CONCLUSIONS: Vitamin D supplementation could improve embryos implantation rates and rate of ongoing pregnancy by decline in the first trimester miscarriage rate.

Role of Dehydroepiandrosterone Supplementation in Improving Intracytoplasmic Sperm Injection Outcome for Women with Expected Poor Ovarian Response

Background: As regard to adjuvant supplementations, nowadays dehydroepiandrosterone (DHEA) is widely used all over the world and is considered to be a potential agent to ameliorate the assisted reproduction technologies outcomes of infertile women with poor ovarian reserve. Objective: To find out the role of DHEA supplementation in improving intracytoplasmic sperm injection (ICSI) outcome for infertile women with expected poor ovarian response in controlled ovarian stimulation. Setting: Assisted reproduction unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Egypt. Duration: From April 2016 to May 2018. Study Design: A randomized double-blinded controlled trial. Methods: One hundred and forty infertile women with expected poor ovarian response prepared for ICSI procedure were included in this study. Patients were divided into two groups;group I (DHEA group) included 70 patients received 25 mg DHEA 12 weeks prior to ICSI cycle and group II (placebo group) included 70 patients received a placebo. Results: There was a highly statistically significant difference in basal AFC at start of ICSI cycle in group I (who received DHEA supplementation for 12 weeks prior to ICSI procedure) than in group II (13.8 ± 5.3 versus 10.7 ± 4.6 respectively) with P < 0.001. There were mildly statistically significant differences between group I and group II as regard to increase in the number and quality of retrieved oocytes, increased in endometrial thickness, fertilization rate and embryo quality with p value < 0.05 but there was no statistically significant difference between the 2 groups as regard to pregnancy (chemical and clinical) rates (p value > 0.05). Conclusions & Recommendations: DHEA supplementations improved basal AFC, increased the number & quality of oocytes and increased quality of embryos in infertile patients with expected poor ovarian response in ICSI procedure. So DHEA supplementations could be an important adjuvant for infertile women with expected poor ovarian response in ICSI procedure.

Sperm DNA fragmentation does not affect the clinical outcomes in the cumulative transfers of an ICSI cycle along with blastocyst transfers in couples with normozoospermic male patients

Objective:To know whether sperm DNA fragmentation(SDF)affects the clinical outcomes in the cumulative transfers of an intracytoplasmic sperm injection(ICSI)cycle along with blastocyst transfers in couples with normozoospermic males.Methods:The study included 252 couples who underwent their first ICSI cycles along with blastocyst transfer and whose male partner semen samples were normozoospermic according to the World Health Organization 2010 criteria.All the couples were classified into two groups based on the SDF:the low SDF group(SDF≤30%,n=162)and the high SDF group(SDF>30%,n=90).Clinical as well as laboratory outcomes were correlated between the two groups.Sperm DNA fragmentation was assessed on the post-wash semen samples by acridine orange test.The main outcome measures were the live birth rate and miscarriage rate.Results:A significant decrease in the live birth rates was observed in the high SDF group compared to the low SDF group in fresh embryo transfer cycles(P<0.05).However,no significant difference was observed in the clinical outcomes either in the frozen embryo transfer cycles or in the overall cumulative transfer cycles(P>0.05).No significant difference was observed in the laboratory outcomes between the two SDF groups.A remarkable decrease in sperm motility was observed in the high SDF group compared to the low SDF group(P<0.05).Conclusions:Sperm DNA fragmentation does not affect the clinical outcomes in the cumulative transfers of an ICSI cycle along with blastocyst transfers in couples with normozoospermic males.

Rejuvenation of Ovary and Thin Endometrium by Autologous PRP Injection in POR and Recurrent Implantation Failure

In India, the problem of infertility is growing and in the last 5 years, it has gone up to 20% - 30%. This ongoing prospective clinical study brings forth a novel, innovative, effective, simple, affordable, easily performed outpatient procedure (OP) and a promising therapeutic method in rejuvenating the Ageing Ovaries and Thin Endometrium, with autologous Platelet Rich Plasma (PRP). This clinical study proves to give a better result in rejuvenating Ovary and treating the Thin Endometrium. This pilot study included five women (28 - 44 years) with Poor Ovarian Response (POR), Premature Ovarian Insufficiency (POI) and Perimenopause and Thirty-nine women (22 - 43 years.) with recurrent implantation failure due to Thin Endometrium were subjected to autologous PRP instillation under Ultrasound Guidance, and Hysteroscopic guided PRP. After PRP, a significant output was obtained, with improved Anti Mullerian Hormone (AMH) and Antral Follicle Count (AFC) and out of five women three women conceived by Intra Cytoplasmic Sperm Injection (ICSI). PRP injected in women with Poor Ovarian Response found successful ovarian rejuvenation within 1 - 3 months and had a 60% of pregnancy rate, PRP into the endometrium had 53.8% successful pregnancies. We have not encountered any complications.

ICSI精子遗传和表观遗传学缺陷

卵胞浆内精子注射(Intracytoplasmic sperm injection,ICSI)技术可用于男性少精、弱精、精子畸形、无精子和常规体外受精周期失败等,克服了精子数量不足甚至直接从附睾、睾丸获取精子来治疗不育。该技术直接将单个精子注射入卵子,因违背自然受精的生物学法则而具有很大的遗传风险。文章对ICSI精子遗传缺陷和表观遗传缺陷及其相关疾病进行综述,可进一步认识ICSI精子遗传与表观遗传缺陷导致后代遗传风险增加的分子的机理,文章阐述了ICSI精子有待于通过DNA甲基化、组蛋白乙酰化等表观遗传因子进行严格质量控制,切实降低ICSI遗传及表观遗传缺陷风险的必要性。

Pureception^(TM)密度梯度离心法优选精子在IVF、ICSI中的应用

目的探讨PureceptionTM密度梯度离心法优选精子在IVF、ICSI中的应用价值。方法采用PureceptionTM密度梯度离心法处理精液,并将获得的精子悬液应用于IVF或ICSI治疗周期。结果精液经处理后,精子密度由(33.86±20.42)×109/L提高到(60.28±41.76)×109/L,活动率由(46.73±10.36)%提高到(85.67±15.31)%,而畸形率则由(30.28±23.44)%降低到(10.49±5.26)%,差异均有显著性(P<0.01)。白细胞及细胞碎片等非精子成分明显减少。87对夫妇共进行119个IVF或ICSI治疗周期,其中42个周期获得临床妊娠,周期临床妊娠率为35.3%。结论PureceptionTM密度梯度离心法是一种较好的精子优选方法,应用于IVF或ICSI可获得较满意的临床妊娠率。

126例梗阻性无精子症借助ICSI治疗的临床结局分析

目的探讨梗阻性无精子症采用不同方式获取的精子对卵胞浆内单精子注射术(ICSI)治疗的妊娠结局。方法根据获取精子的方式不同将梗阻性无精子症患者分为两组:TESA组46例、PESA组80例,两组患者均行ICSI治疗。比较两组受精率、卵裂率、优质胚胎率、种植率及临床妊娠率等。结果 TESA组与PESA组的卵裂率分别为88.2%和94.2%,优质胚胎率分别为62.2%和62.5%,两组比较差异均无统计学意义(P>0.05)。两组受精率分别为72.1%和77.6%,种植率分别为23.3%、34.8%,临床妊娠率分别43.1%、68.7%,两组比较差异均有统计学意义(P<0.05)。结论 PESA或TESA结合ICSI是治疗梗阻性无精子症的有效治疗方法 ,附睾精子优于睾丸精子,可首选抽取附睾精子。

不同精子处理对应用ICSI-Tr技术生产转基因水牛胚胎效率的影响

本文主要探讨了不同精子处理对应用显微授精介导(intracytoplasmic sperm injection-mediated trans-genesis,ICSI-Tr)生产转基因水牛胚胎效率的影响。本试验以p18T-BCN5.2-IFN-1.1pA-EGFP为载体,比较了精子的不同处理方式(NaOH和冻融)、NaOH处理精子不同时间(5min,10min,20min,30min和60min)以及不同外源DNA浓度(5ng/μL,10ng/μL,25ng/μL和50ng/μL)对ICSI-Tr转基因效率的影响。结果显示:NaOH处理组的囊胚EGFP表达率(46.1%),与冻融精子处理组(47.1%)差异不显著,但发育率显著高于冻融处理精子组(28.3%vs16.7%,P<0.05)。处理60min组和30min组的分裂率分别为78.9%和77.5%,显著高于20min、10min和5min组的64.0%、60.0%和64.0%(P<0.05),其中30min处理组的囊胚EGFP表达率高达44.4%,显著高于其它处理组(P<0.05)。25ng/μL组和50ng/μL组的早期胚胎EGFP表达效率差异不显著(41.3%vs42.5%),但显著高于5ng/μL组和10ng/μL组(22.5%和35.5%),25ng/μL组的囊胚EGFP表达效率显著高于其它组(P<0.05)。本研究优化了应用ICSI-Tr生产转基因水牛胚胎技术体系,为获得ICSI-Tr转基因水牛奠定了良好的工作基础。

体外助孕治疗周期中分半进行IVF和ICSI的指征

目的对体外助孕治疗周期中存在不受精可能且上游有效精子数量不低于500万的夫妇分半进行常规体外受精（IVF）与卵胞浆内单精子显微注射（ICSI）治疗，探讨是否存在分半治疗的必要性以及指征选择的可行性。方法2002年3月～2007年6月接受体外助孕的夫妇中选取符合指征周期，采用常规超排卵方案，同一周期取得的卵子随机分半，分别进行常规IVF与ICSI治疗。卵裂球≥5个且卵裂球碎片〈50％的胚胎为可用胚胎。结果共有94周期进入分半治疗周期，其中常规IVF受精失败或低下（LFR）22周期（23．40％）；同期常规IVF治疗530周期，其中LFR36周期（6．79％）；两者有显著差异（P=5．26×10^-6）。94分半周期共获1596枚卵子，其中行常规IVF781枚，受精率55．19％；行ICSI815枚，MⅡ616枚，受精率72．02％；两者有显著差异（P=2．59×10^-12）。94周期卵裂983枚，可用胚胎554枚，临床妊娠率为35．7l％。结论存在同一治疗周期分半进行常规IVF与ICSI的指征和必要性，可明显减少不受精情况的出现；对常规IVF与ICSI均受精的周期，可选择移植常规IVF胚胎，减少子代遗传和先天缺陷风险。

短时受精结合早期补救ICSI技术的临床应用

常规授精中受精失败已成为辅助生殖技术面临的严峻问题,目前常用的补救措施有Half-ICSI和短时受精结合早期补救ICSI。短时受精结合早期补救ICSI技术在挽救IVF受精失败周期中有很多优势,已经被很多生殖中心所采用。但是关于短时受精对胚胎发育及临床妊娠率的影响仍存在很多争议,此外早期补救ICSI的有效性也是制约该技术应用的一个障碍。本文综述了短时受精结合早期补救ICSI技术的进展以及影响其成功率的因素,希望为该技术的临床应用提供参考。

99例分半IVF/ICSI指征及临床结局的分析与探讨

目的:探讨可疑体外受精(IVF)受精失败的患者在体外助孕治疗周期中如何选择合理的受精方式。方法:回顾性分析2002年1月至2007年10月在本中心接受同胞卵母细胞分半IVF/ICSI(half-ICSI)治疗的99个周期,促排卵后共获卵1791个,其中895个卵行IVF,另896个卵中有753个MⅡ期行ICSI,比较不同受精方式的受精率、卵裂率、优胚率及妊娠率。结果:half-ICSI中ICSI受精率(80.5%)明显高于IVF受精率(42.9%),IVF完全受精失败率(21.2%)及受精低下率(16.2%)明显高于ICSI完全受精失败率(0)及受精低下率(3.0%),差异有统计学意义(P<0.01);IVF和ICSI的卵裂率、优胚率比较无显著性差异(P>0.05)。结论:对可疑IVF受精失败的患者,在患者充分知情同意下,选择ICSI是必要的,可提高受精率,获得足够优质的胚胎移植,增加妊娠机会。

拮抗剂方案OPU-ICSI不同时机选择对新鲜胚胎移植周期临床结局的影响

目的:比较促性腺激素释放激素(GnRH)拮抗剂方案中选择不同的胞浆内单精子注射(ICSI)实施时机对新鲜胚胎移植周期中胚胎质量及妊娠结局的影响。方法:选择2016年1月—2018年5月在我科采用GnRH拮抗剂方案促排卵,行新鲜胚胎移植的531个周期,按不同取卵(OPU)-ICSI间隔时间分为2组:38~40h为组1(n=101),41~43h为组2(n=430),比较2组患者的卵母细胞成熟度、胚胎质量和妊娠结局。结果:组1患者的卵子成熟率低于组2,但组2的早期流产率低于组1,差异有统计学意义(均P<0.05)。2组患者的受精率、分裂率、可利用胚胎率、优胚率、种植率和临床妊娠率等比较,差异无统计学意义(均P>0.05)。结论:在GnRH拮抗剂方案新鲜胚胎移植周期中,ICSI时机选择在38~43h的窗口期均能达到比较一致的胚胎质量及临床妊娠率。关于OPU-ICSI间隔短(小于40h)早期流产率较高,需要更多的和细致分组的研究。

卵母细胞胞浆内精子注射(ICSI)技术的研究进展

利用人工辅助的显微操作技术,直接将精子注射至成熟卵母细胞质的方法,称为胞浆内精子注射技术,简称ICS I(in tracytop lasm ic sperm in jection)技术。目前,该项技术已经成功地用于生产试管婴儿、研究哺乳动物的受精机理、动物转基因、家畜性控胚胎生产等领域。本文详述了ICS I技术体系的发展背景、现状及其应用等方面。

卵子第一极体、卵周隙及胞浆形态与ICSI后受精率和胚胎质量的影响

目的 从MII期卵子的第一极体、卵周隙和胞浆形态三方面综合探讨卵子质量对卵母细胞浆内单精子注射(ICSI)后的受精率、卵裂率、优胚获得率的影响。方法 对ICSI中去除颗粒细胞后的MII期卵子在光镜下从第一极体、卵周隙和胞浆形态三方面综合评价卵子质量并按形态表现分为五级。且分别比较各级卵子的受精率、卵裂率及优胚率。结果 Ⅰ级卵子与其他各级含异常形态的卵子比较 ,受精率、卵裂率和优质胚胎率 ,均有显著差异 (P <0 .0 1)。Ⅰ级卵子有最高的受精率 (91.14 % )、卵裂率 (96 .35 % )和优质胚胎率 (83.91% )。形态异常的卵子的受精率、卵裂率和优质胚胎率均有明显的下降趋势。在取卵数≤ 15个及hCG注射日每个卵泡的血清E2浓度在 732～ 10 98pmol/L组中 ,可获得高比例的Ⅰ级卵子。结论 根据MII期卵子的第一极体、卵周隙及胞浆形态三方面综合因素进行的卵子分级是可预测ICSI后受精率、卵裂率及优胚率。

显微操作时间对猪ICSI胚胎早期发育的影响

为了进一步优化现有的猪卵胞质内精子注射(ICSI)体系,探讨了ICSI操作时间对猪ICSI胚胎早期发育的影响情况。结果表明,囊胚形成率随着ICSI操作时间的延长而呈现明显下降趋势,其中卵母细胞在体外4 h内完成ICSI组的囊胚形成率(5.90%)显著低于其他3个处理(P<0.01)。可见,ICSI操作时间越长,其胚胎早期发育率越低。

不同取卵时间对兔ICSI胚胎体外发育的影响

目的探讨不同取卵时间对兔ICSI胚胎体外发育的影响。方法采用Piezo操作系统对实验兔进行辅助体外受精。结果hCG注射后14、161、8 h取卵,ICSI后的受精率分别为82.2%、75.9%和0.0%,对受精卵进行体外发育培养,桑椹胚的发育率分别为72.9%、70.0%、0.0%,囊胚的发育率分别为62.2%、53.3%、0.0%。14 h和16 h之间受精率、桑椹胚率、囊胚率差异不显著(p>0.05),但是14 h采卵比16 h要好;18 h和14 h1、6 h之间差异显著(P<0.05)。结论不同取卵时间影响实验兔的ICSI体外受精率及胚胎的体外发育率,hCG注射后14 h取卵最有利于兔ICSI胚胎的发育。

ICSI子代与自然妊娠儿童学龄前期体格及智力发育比较分析

目的了解卵胞浆内单精子注射（ICSI）技术对子代学龄前期体格及智力发育的影响。方法使用1：1病例对照研究方法,回顾性随访收集嘉兴市妇幼保健院生殖医学中心行ICSI技术怀孕成功,并在2009年1月至2010年12月间分娩的儿童病例信息,用电话召回,将愿意参加研究的家庭作为ICSI组。在同期本院出生自然怀孕儿童作为对照组。随访家庭一般情况,出生信息,学龄前期体格发育情况,同时进行韦氏学龄前智力测试。结果 ICSI组及对照组各收集到30例。两组均为男16人,女14人,其中ICSI组双胎3对,对照组无。母亲生育年龄,母亲文化程度,父亲生育年龄,父亲文化程度,均未见显著性差异（均P〉0.05）。两组分娩方式,出生体重均值,4~6岁身高与体重均与对照组无显著性差异（均P〉0.05）。仅出生孕周均值ICSI组低于对照组（37.27±2.392 vs 38.80±1.349）,χ2=3.058,P=0.003。韦氏学龄前智力测试结果比较,仅言语商中的图片概括得分ICSI组显著低于对照组（11.00±3.283 vs 13.92±3.283）,χ2=2.893,P=0.006。其余各项得分和总智商两组间均无显著性差异（均P〉0.05）。结论随访至学龄前期,ICSI子代的体格发育和智力发育情况在正常范围,与自然妊娠儿童发育情况接近。

附睾穿刺取精行ICSI治疗梗阻性无精子症

目的探讨附睾穿刺取精术(PESA)结合单精子卵胞浆内注射(ICSI)治疗梗阻性无精子症男性不育的可行性,并观察其临床效果。方法 7对夫妇为研究对象,男方均确诊为梗阻性无精子症,女方超促排卵获得卵细胞,男方于取卵日在局麻下通过细针穿刺附睾头部吸取少量精子,行ICSI,受精成功后24-48h,选择优质胚胎移植入子宫腔。因男性少弱精子症行ICSI治疗的20个治疗周期为对照组。结果附睾取精7例共11个治疗周期全部获得活动精子,ICSI后受精率65．9％,卵裂率98．3％,优质胚胎率71．9％,临床妊娠5例,周期临床妊娠率45．5％,与对照组比较,各项指标均无显著差异。结论附睾穿刺取得的精子与排出体外的精子具有相同的受精和获得优质胚胎的能力,PESA是治疗梗阻性无精子症男性不育的安全有效的方法。

牛分离精子ICSI后卵母细胞激活方法的研究

本研究的目的是比较采用由流式细胞仪分离的牛X和Y精子进行胞质内精子注射(ICS I)后,两种不同激活方法对ICS I卵母细胞的激活效果。ICS I后,卵母细胞用5μm o l/L的Ionom yc in(离子霉素)处理5 m in后,先在化学成分明确培养液(CDM-1)中培养3 h,然后再在含有1.9 mm o l/L 6-DM AP(6二-甲氨基嘌呤)的培养液中培养3 h(Ionom yc in+CDM-1+DM AP),或用5μm o l/L的Ionom yc in处理5 m in后,不经过CDM培养直接转入1.9 mm o l/L DM AP的培养液中培养3 h(Ionom yc in+DAM P)。结果发现,两种激活处理的效果差异不显著。

IVF/ICSI-ET/FET后自然流产的胚胎染色体异常的相关因素分析

目的探讨体外受精(in vitro fertilization,IVF)、卵泡浆内单精子注射(intracytoplasmic sperm injection,ICSI)后新鲜胚胎移植(embryo-transfer,ET)及冷冻胚胎移植(frozen embryo transfer,FET)后自然流产的胚胎染色体异常的相关危险因素。方法选择2008年10月至2012年4月于南方医科大学南方医院生殖中心行IVF/ICSI-ET/FET后发生自然流产的夫妇115例,根据胚胎染色体结果分为异常组(74例)及正常组(41例),比较两组的一般资料及助孕相关指标间的差异并探讨胚胎染色体异常的相关危险因素。结果IVF/ICSI-ET/FET中自然流产绒毛染色体异常率为64.3%,以非整倍体率(91.9%)为主。胚胎染色体异常组的女性年龄显著高于正常组[(34.3±4.5)vs(32.2±4.5),P<0.05],促性腺激素(Gonadotrophin,Gn)总量显著高于后者[(2935.5±1418.5)vs(2398±1014.9),P<0.05],男性年龄亦高于后者[(37.1±4.9)vs(35.3±5.2),P>0.05],但差异无统计学意义。正常组的男女性别比为1∶2.4,女胎比例显著高于异常组(P<0.05)。IVF与ICSI的胚胎染色体异常类型差异无统计学意义(P>0.05)。Logistic回归分析提示仅女性年龄为胚胎染色体异常的独立危险因素(OR:1.008~1.204)。结论胚胎染色体异常是IVF/ICS-ET/FET后自然流产的主要原因,但细胞培养法可能会降低胚胎染色体异常的检出率。女性年龄是胚胎染色体异常的独立危险因素,而IVF/ICS-ET/FET治疗并不增加胚胎染色体异常的风险。