目的  探讨精子DNA碎片指数（DFI）对体外受精-胚胎移植（IVF-ET）胚胎发育及妊娠结局的影响，为优化男性生育力评估及辅助生殖治疗策略提供依据。方法  回顾性分析2023年1月—2024年1月于徐州市妇幼保健院接受IVF-ET治疗的126对不孕夫妇，根据男方DFI检测结果分为低碎片组（DFI≤15%，n=42）、临界组（15%＜DFI＜30%，n=45）和高碎片组（DFI≥30%，n=39）。比较三组患者受精相关指标、胚胎发育指标及妊娠结局指标的差异，并分析DFI与各指标的相关性。结果  低碎片组双原核率（2PN）率、优质胚胎率及囊胚形成率均高于临界组和高碎片组（P＜0.001），低碎片组1PN率、多PN率均低于临界组和高碎片组（P＜0.001）；三组种植率、临床妊娠率、早期流产率比较差异无统计学意义（P＞0.05），但高碎片组活产率低于低碎片组（P＜0.05）。相关性分析结果表明，DFI与优质胚胎率（r=-0.412，P＜0.001）、囊胚形成率（r=-0.387，P＜0.001）、活产率（r=-0.287，P=0.012）呈负相关，与早期流产率（r=0.206，P=0.059）、种植率（r=-0.215，P=0.058）、临床妊娠率（r=-0.203，P=0.072）无显著相关性。结论  精子DNA碎片指数是影响IVF-ET胚胎发育及妊娠结局的重要因素，高DFI主要通过降低胚胎发育潜能及增加流产风险导致活产率下降，临床需对高DFI患者进行干预以改善治疗结局。

       Objective  To investigate the impact of sperm DNA fragmentation index（DFI）on embryo development and pregnancy outcomes of in vitro fertilization-embryo transfer（IVF-ET），and to provide a basis for optimizing male fertility assessment and assisted reproductive treatment strategies．Methods  A retrospective analysis was performed on 126 infertile couples undergoing IVF-ET treatment at the Reproductive Medicine Center of Xuzhou Maternal and Child Health Hospital from January 2023 to January 2024．According to the male DFI test results，they were divided into three groups：low fragmentation group（DFI≤15%，n=42），critical group（15% ＜ DFI ＜ 30%，n=45），and high fragmentation group（DFI≥30%，n=39）．Differences in fertilization-related indicators，embryo development indicators，and pregnancy outcome indicators were compared among the three groups，and the correlation between DFI and each indicator was analyzed．Results  The 2 pronuclei rate（PN）rate，high-quality embryo rate，and blastocyst formation rate in the low fragmentation group were significantly higher than those in the critical and high fragmentation groups（P＜0.001）．The 1PN  rate and multi-PN  rate in the low fragmentation group were significantly lower than those in the critical and high fragmentation groups（P＜0.001）．There was no significant difference in the three groups of implantation rate，clinical pregnancy rate and early abortion rate（P＞0.05），but the live birth rate of high fragment group was significantly lower than that of low fragment group（P＜0.05）．The results of correlation analysis showed that DFI was significantly negatively correlated with the rate of high quality embryos（r=-0.412，P＜0.001），blastocyst formation rate（r=-0.387，P＜0.001）and live birth rate（r=-0.287，P=0.012），but not with the rate of early abortion（r=0.206，P=0.059），implantation rate（r=-0.215，P=0.058）and clinical pregnancy rate（r=-0.203，P=0.072）．Conclusions  Sperm DFI is an important factor affecting embryo development and pregnancy maintenance in IVF-ET．High DFI leads to a decrease in live birth rate mainly by reducing embryo developmental potential and increasing the risk of early abortion．Clinically，early intervention is needed for patients with high DFI to improve treatment outcomes．

目的 比较来曲唑和氯米芬治疗极严重少精子症的治疗效果。方法 选取2020—2021年在本生殖中心门诊50例极严重少精子症病例,分为2组,A组(来曲唑,研究组n=26)、B组(氯米芬,对照组n=24)。结果 来曲唑和氯米芬治疗后1月、3月,A组精子浓度明显提高,优于B组[(4.5±3.1) vs (2.0±1.3);(8.3±3.5) vs (2.0±1.8),P<0.05]。通过卵胞浆内单精子注射/体外受精胚胎移植治疗,A组妊娠16例(61.5%),B组妊娠11例(45%)。2组治疗后的血清卵泡刺激素、黄体生成素、睾酮升高[(5.9±3.8) vs (20.3±2.6);(3.6±2.8) vs (9.5±5.7);(13.6±10.5) vs (25.3±10.8),P<0.05],A组雌二醇水平降低[(36.8±20.6) vs (7.6±2.5),P<0.05]。结论 来曲唑治疗极严重少精子症有较理想的临床治疗效果,明显优于氯米芬。

Objective To explore and compare the therapeutic effects of letrozole and clomiphene in the treatment of extremely severe oligospermia. Methods A tatal of 50 cases of extremely severe oligosperospermia in the reproductive center from 2020 to 2021 were selected and divided into two groups, group A (letrozole, study group n=26) and B (clomiphene, control group n=24). Results After letrizole and clomiphene treated for 1 and 3 months, sperm concentration in group A was significantly better than group B[(4.5±3.1) vs (2.0±1.3);(8.3±3.5) vs (2.0±1.8),P<0.05]. With ICSI/IVF, 16 cases (61.5%) succeed in encyesis in group A, 11 cases (45%) in group B. Serum follicle stimulating hormone, luteinizing hormone and testosterone levels were significantly increased after treatment [(5.9±3.8) vs (20.3±2.6), (3.6±2.8) vs (9.5±5.7), (13.6±10.5) vs (25.3±10.8), P<0.05]. The level of estradiol in group A decreased [(36.8±20.6) vs (7.6±2.5), P<0.05]. Conclusions Letrozole has ideal clinical treatment effect, which is obviously superior to clommiphene.

目的 通过对不同受孕能力精子外显子的分析,寻找并验证特异性外显子作为精子受孕能力的生物标记物。 方法 基于二代测序数据进行生物信息学分析,寻找特异性外显子并设计引物。各取8份高、低受孕能力精液标本,提取精子RNA反转录后进行real time q-PCR验证外显子的表达效率,筛选表达差异恒定的精子外显子作为生物标记物。各取10份高、低受孕能力精子标本,用筛选后的外显子引物进行real time q-PCR验证。 结果 生物信息学分析得到31个候选精子外显子,从31个候选外显子中筛选出9个表达差异恒定的精子外显子GAPDHS、HSF2BP、HSPA1L、ADAM21、SPEM1、WBP2NL、DDX20、TSGA10、PGK2;real time q-PCR验证结果显示,在高、低受孕能力精液标本中这9种精子外显子表达差异明显。 结论 初步确定,差异表达恒定的九种外显子可作为评估精子质量的生物标记物。

Objective To find and verify specific exons as biomarkers of sperm fertility by analyzing sperm exons with different fertility ability.Methods Based on the second generation sequencing data, bioinformatics analysis was conducted to find specific exons and design primers. We obtained 16 semen samples, 8 of high and the other 8 of low fertilizing ability, after the sperm RNAs were extracted and reverse-transcribed, real time q-PCR was performed to verify the expression efficiency of exons, and the sperm exons with constant expression difference were selected as biomarkers. 10 high and 10 lowfertility ability sperm samples were taken for real time q-PCR verification with screened exon primers. Results Thirty-one sperm exons were obtained by bioinformatics analysis, and 9 sperm exons with constant expression differences were selected from the 31 candidate exons, including GAPDHS, HSF2BP, HSPA1L, ADAM21, SPEM1, WBP2NL, DDX20, TSGA10 and PGK2. The results of real time q-PCR verification showed that the exons of these 9 sperm were significantly different in the semen samples with high and low fertility ability. Conclusion Nine exons with constant differential expression can be used as biomarkers to evaluate sperm quality.

目的 探讨一次不明原因流产与精子DNA损伤的关系。方法 收集有一次不明原因流产史的患者作为实验组,同时以有正常妊娠史的患者为对照组,分别比较两组男方年龄、精子密度、精子活力、精液量和精子DNA断裂指数有无差异。以SPSS 16.0为统计软件,进行独立样本的t检验。结果 两组的精液量、精子密度及活力均无差异,实验组男方年龄小于对照组(P<0.05),但实验组的DFI要高于对照组(P<0.01)。结论 本研究对照组年龄高于实验组,而DFI正好相反。这说明不明原因的自发流产与男方精子DFI密切相关,随着DFI的增加,流产风险增加。

Objective To investigate the relationship between unexplained abortion and sperm DNA damage. Methods Patients with a history of unexplained abortion were enrolled as an experimental group, and patients with a normal pregnancy history were used as a control group. The differences in age, sperm density, sperm motility, semen volume, and sperm DNA break index were compared between the two groups. The independent sample t test was performed using the statistical software of SPSS 16.0. Results There was no significant difference in semen volume, sperm density and motility between the two groups. The age of the male group in the experimental group was lower than that in the control group (P<0.05), but the DFI of the experimental group was higher than that of the control group, and the difference was statistical significance (P<0.01). Conclusion The age of the study control group was higher than that of the experimental group, while DNA fragmentation index DFI was the opposite, which indicated that unexplained spontaneous abortion was closely related to the male sperm DFI, and the risk of miscarriage may increase with the increase of DFI.

目的 探讨精液白细胞和形态正常精子百分率的相关性。方法 随机选取2014年4月—2015年8月来生殖中心进行就诊的男性800例,分析患者精液中的白细胞数量以及分析患者的精子形态,比较二者的相关性。结果 正常形态精子百分率低于正常参考值的患者中有61.9％的白细胞数大于1×10⁶,精子形态正常的患者有38.9％患者的白细胞数大于1×10⁶,患者比例差异有统计学意义(P<0.05)。精子形态正常的患者(320例)和正常形态精子百分率低于正常参考值的患者(480例)相比,精子形态正常的比例远远大于正常形态精子百分率低于正常参考值的患者(P<0.05)。而正常形态精子百分率低于正常参考值的患者的头部异常、颈部和中间部分异常、尾部异常精子比例高于正常组(P<0.05)。正常形态精子百分率低于正常参考值的和正常形态的精子比例呈正相关;和大头精子百分率、锥形以及小头形态精子百分率呈负相关;和梨形以及其他形态的精子呈正相关。结论 精液中的白细胞可以影响精子的参数,使得患者精液中的颈部或中部异常形态、头部和尾部异常形态的精子比例提升,具体机制有待进一步研究。

目的 探讨彩色多普勒超声在无精子症患者睾丸穿刺活检中的应用价值。方法 回顾性分析2012年11月—2015年2月在我科诊断为无精子症的53例患者的病例资料。所有患者进行常规阴囊超声检查,采用频谱多普勒检测睾丸动脉(TA)、睾丸内动脉(ITA)的收缩期最大速度(PSV)、舒张末期速度(EDV)和阻力指数(RI)。采用彩色多普勒超声观察睾丸内的血管分布及其数目,并进行半定量评分。所有患者进行睾丸穿刺活检并进行Johnsen评分。结果 47例患者诊断为梗阻性无精子症(OA),6例患者诊断为非梗阻性无精子症(NOA)。0A组睾丸的平均体积(15.3±3.6) mL,NOA组睾丸的平均体积(7.1±2.8) mL,差异有统计学意义(P＜0.01);OA组与NOA组患者睾丸的血流参数(PSV、EDV、RI) 结果有统计学意义(P＜0.05);OA组与NOA组睾丸内血管半定量分级有的差异,其差异有统计学意义(P＜0.01)。结论 彩色多普勒超声检查对睾丸生精功能有很好的预测作用,可作为鉴别OA患者和NOA患者的指标。

Objective To explore the application value of color Doppler ultrasound in patients with azoospermia who underwent testicular needle biopsy. Methods Retrospective analysed the clinical data of 53 cases patients with azoospermia from 2012 November to 2015 February in our department.All patients underwent routine scrotal ultrasonography, to detecte the peak systolic velocity (PSV), end diastolic velocity (EDV) and resistance index (RI) of testicular artery (TA)and intratesticular artery (ITA)with spectral Doppler ultrasound. Color Doppler ultrasound was used to observe the distribution and the number of blood vessels in the testicle, and semi quantitative score. All patients underwent testicular biopsy and Johnsen score. Results 47 patients were diagnosed as obstructive azoospermia(OA), 6 patients were diagnosed as non obstructive azoospermia(NOA). The average volume of testis was (15.3±3.6)mL in 0A group. The average volume of testis was (7.1±2.8) mL in N0A group.There was significant difference (P<0.01). The blood flow parameters(PSV, EDV, RI) results between OA group and NOA group have obvious difference,the difference was statistically significant (P<0.05). The semi quantitative classification of testicular vessels between OA group and NOA group have obvious difference,the difference was statistically significant (P< 0.01). Conclusion Color Doppler ultrasound is a good predictor of testicular spermatogenic function, can be used as the identification index of patients with OA and patients with NOA.

目的 分析影响接受夫精宫腔内人工授精(IUI)助孕患者临床妊娠率的有关因素。方法 选择225例接受IUI助孕共400周期的不孕症患者,回顾分析女方年龄、治疗方案、hCG日子宫内膜厚度、IUI周期数与临床妊娠率的关系。结果 女方年龄>38岁的临床妊娠率(8.22%)低于年龄<30岁(21.74%)和30~38岁(17.48%),P<0.05;促排卵方案(CC、HMG、CC+HMG)临床妊娠率分别为19.05%、19.66%和14.71%,高于自然周期的临床妊娠率7.14%,P<0.05;hCG日子宫内膜厚度≥8 mm组的临床妊娠率(23.56%)高于内膜<8 mm组的临床妊娠率(13.27%),P<0.05;第1至第5周期的IUI临床妊娠率分别为21.30%、15.60%、9.38%、0%和0%,多次重复IUI周期数差异有统计学意义(P<0.05)。结论 女方年龄、hCG日子宫内膜厚度、治疗方案均会影响IUI的临床妊娠率,但增加IUI的治疗周期数并不能提高临床妊娠率,应综合各种因素再次评估患者的妊娠率,必要时进一步查找多次助孕失败的原因或改行IVF-ET助孕治疗。

Objective To analyze the relative factors which influence the clinical pregnancy rates of patients accepted intrauterine insemination with husband's sperm. Methods 225 cases of infertile patients accepted IUI treatment were selected, 400 cycles were included and the clinical data were analyzed retrospectively. Observing the relationship between the age of women, treatment options, endometrial thickness on hCG injection day, cycles of IUI and pregnancy rates. Results The clinical pregnancy rates of women less than 30 years old(21.74%) were higher than aged between 30 to 38 years old(17.48%) and more than aged 38 years old(8.22%), P<0.05.The clinical pregnancy rates of ovulation induction options(CC、HMG、CC+HMG) were 19.05%, 19.66% and 14.71%, higher than the pregnancy rates of natural cycle 7.14% significantly, P<0.05. The pregnancy rates of the group of endometrial thickness ≥8mm on hCG injection day were 23.56%, higher than the group of endometrial thickness <8 mm 13.27%, P<0.05. The clinical pregnancy rates of 1 to 5 cycles IUI were 21.30%, 15.60%, 9.38%, 0% and 0% respectively, the difference of repeating the IUI cycles’ number was statistical significance (P<0.05). Conclusion The ages of women, endometrial thickness on hCG injection days and treatment options can affect the clinical pregnancy rates. Extending the number of IUI treatment cycles can not increase the pregnancy rates of IUI. All the factors should be comprehensive to assess the patient's pregnancy rates again, to find more reasons further for the failure of assisted reproduction or turn to IVF-ET assisted reproduction treatment when it is necessary.

目的 观察十一酸睾酮联合左卡尼汀治疗特发性弱精症的疗效。方法 回顾性分析2012年11月—2014年6月在我科诊断为弱精子症的386例患者的临床资料,按标准分成轻(A组)、中(B组)、重(C组)3组。分别给予左卡尼汀、十一酸睾酮联合左卡尼汀治疗,观察两种治疗方法对特发性弱精子症的疗效。结果 A组、B组患者两种治疗方法治疗后精子PR均有升高,差异有统计学意义(P＜0.05),C组患者给予左卡尼汀治疗患者治疗后PR无改善,差异无统计学意义(P＞0.05);十一酸睾酮联合左卡尼汀治疗患者治疗后PR升高,差异有统计学意义(P＜0.05)。A、B、C组患者给予左卡尼汀治疗的有效率分别是47.6%,41.7%,18.9%,总有效率为37.9%;A、B、C组患者给予十一酸睾酮联合左卡尼汀治疗的有效率分别是52.6%,45.5%,34.7%,总有效率为45.5%。A组、B组患者两种治疗方法疗效无差异(P＞0.05);C组患者给予十一酸睾酮联合左卡尼汀组有效率高于给予左卡尼汀组,差异有统计学意义(P＜0.05)。结论 十一酸睾酮联合左卡尼汀治疗特发性弱精子症的疗效优于左卡尼汀单一用药。

Objective To observe the effects of eleven acid testosterone combined with L-carnitine on curing idiopathic asthenospermia. Methods Retrospective analysed the clinical data of 386 patients with asthenospermia patients from November 2012 to June 2014 in our department.According to the standard,the patients were divided into mild A,moderate B,severe C in 3 groups,given L-carnitine treatment and given eleven acid testosterone combined with L-carnitine treatment.We observed effect of the two therapeutic methods for the treatment of idiopathic asthenospermia. Results The sperm PR were increased,In group A and group B patients after treated.The difference was statistically significant(P<0.05).In group C,the patients received the treatment of L-carnitine,the sperm PR has no obvious improvement,There was no significant difference(P>0.05).The patients received the treatment of eleven acid testosterone combined with L-carnitine the sperm PR significantly increased,the difference was statistically significant(P<0.05).The treatment efficiency of A,B,C group received L-carnitine were 47.6%,41.7%,18.9%,the total effective rate was 37.9%.The treatment efficiency of A,B,C group received eleven acid testosterone combined with L-carnitine were 52.6%,45.5%,34.7%,the total effective rate was 45.5%;There was no significant difference in group A and group B(P>0.05).The treatment efficiency in group C received eleven acid testosterone combined with L-carnitine group is obviously higher than that given L-carnitine group,the difference was statistically significant(P<0.05). Conclusion The curative effect of eleven acid testosterone combined with L-carnitine in the treatment of idiopathic asthenospermia is better than that of L-carnitine single drug treatment.