男性不育治疗技术的研究进展

卵细胞浆内单精子显微受精（intracytoplasmic sperm injection,ICSI)^[1]技术的成功应用，给男性不育的治疗带来了革命性的突破，近几年ICSI技术不断改进和完善，出现了外科手术法从睾丸和附睾取精（testicular sperm extraction,TESE)^[2,3]与ICSI相结合技术（TESE－ICSI）^[4]，睾丸和附睾抽吸精子（testicular sperm aspiration,TESA)与ICSI相结合技术（TESA－ICSI）^[5,6]以及TESE－ICSI与冷冻精子相结合技术^[7]。尽管男性不育治疗技术在不断完善和成熟，但是仍然有许多问题还未解决。     

睾丸容积、染色体及血清生殖激素对无精子症睾丸内精子存在的评估作用

无精子症,在男性不育患者中占5%～20%,包括染色体畸变、内分泌激素异常等病因[1]。无精子症病因复杂、临床治疗困难,是导致男性绝对不育的重要原因。临床上常采用经皮附睾精子抽吸术（percutaneous epididymal sperm aspira-tion,PESA）和睾丸精子抽吸术（testicular sperm extraction,TESE）,通过手术获取成熟精子行辅助生殖技术。     

睾丸与附睾精子对梗阻性无精子症ICSI助孕结局的影响

目的比较睾丸精子与附睾精子对梗阻性无精子症(obstructive azoospermia,OA)患者行卵胞浆内单精子注射(intracytoplasmic sperm injection,ICSI)治疗结局的影响。方法收集2014年1月至2016年12月在海南医学院第一附属医院生殖医学中心因梗阻性无精子症行ICSI助孕治疗患者的临床资料,共163个周期。根据精子来源分为两组,TESA组:采用睾丸精子抽吸术(testicular sperm aspiration,TESA)取精,共137个周期;PESA组:采用经皮附睾精子抽吸术(percutaneous epididymal sperm aspiration,PESA)取精,共26个周期。比较两组的正常受精率、2PN卵裂率、优质胚胎率、胚胎利用率、胚胎种植率、临床妊娠率、流产率及活产率之间有无差异。结果 TESA组的胚胎种植率、临床妊娠率及活产率高于PESA组,但差异均无统计学意义(P0.05);TESA组的正常受精率、2PN卵裂率、优质胚胎率、胚胎利用率及流产率均低于PESA组,但差异亦无统计学意义(P0.05)。结论对梗阻性无精子症患者,睾丸精子和附睾精子ICSI后可达到相似的助孕结局。     

睾丸精子抽吸后行卵细胞单精子显微注射治疗梗阻性无精子症

无精子症约占男性不育患者的7％～14％。目前尚无确切的治疗方法。自Palerm等于1992年首次报道卵细胞浆内单精子注射（ICSI）获得妊娠成功，为男性不育症治疗提供了新方法，同时附睾或睾丸取精术彻底改变了无精子症不可治疗的局面。采用此方法只要在男性生殖道或睾丸内发现并分离到精子，利用ICSI技术就能获得一定的妊娠率，     

采用抽吸取精法结合单精子注射治疗无精子症结局

目的评价经皮附睾精子抽吸术（percutaneus epididymal sperm aspiration,PESA）或睾丸精子抽吸术（testicular sperm aspiration,TESA）结合卵胞浆内单精子注射（intracytoplasmic sperm injection,ICSI）治疗无精子症的临床效果。方法对290例因男性梗阻性及非梗阻性无精子症（non-obstructive azoospermia,NOA）采用PESA或TESA穿刺获取精子,女方采用长方案超排卵,然后对处于细胞分裂中期的成熟卵母细胞进行单精子注射。结果梗阻性无精子症组203例,受精率77.5%,临床妊娠率46.1%;非梗阻性无精子症组87例,受精率73.0%,临床妊娠率41.4%,两组比较其受精率及临床妊娠率均无显著性差异（P〉0.05）。结论采用PESA或TESA获取精子结合ICSI是治疗梗阻性及非梗阻性无精子症等严重的男性不育症的一种有效的方法。     

采用经皮抽吸取精法结合卵胞浆内单精子注射治疗无精子症52例临床报道

在不孕不育夫妇中,由男性因素导致不育的夫妇占40%。自1992年Palermo等[1]首次用卵胞浆内单精子注射术（intra-cytoplasmic sperm injection,ICSI）治疗男性少,弱精子症的有效方法以来,使无精症等疾病的患者也有了生育后代的希望。近几年来,由于ICSI技术的不断发展和提高,以治疗男性不育     

睾丸精子与附睾精子结合单精子注射治疗无精子症妊娠结局比较

目的比较附睾精子和睾丸精子结合单精子注射(ICSI)治疗无精子症的妊娠结局。方法 216例无精子症患者经皮附睾穿刺抽吸术取得附睾精子;74例无精子症患者经皮睾丸精子抽吸术(TESA)获得睾丸精子。女方进行常规超排卵。采用卵胞浆内单精子注射技术获得妊娠,比较两者的受精率、种植率和临床妊娠率。结果附睾精子组和睾丸精子组的受精率分别为75.20%和74.61%,比较其差异无显著性(P0.05);两者的种植率和临床妊娠率分别为29.18%vs 23.89%和52.43%vs 40.21%,差异具有显著性(P0.05)。结论附睾是精子获能、成熟的重要部位,附睾精子优于睾丸精子,对无精子症患者行ICSI之前尽可能首先选取附睾精子。     

改良卵胞浆内单精子显微注射技术在男性不育中的应用

目的 探讨改良卵胞浆内单精子显微注射技术(intracytoplasmic sperm injection,ICSI)及其对男性不育的治疗效果。方法 对ICSI进行改良及应用此技术对97例不孕不育患者、女方常规超促排卵、男方采用手淫(91例)、睾九穿刺取精(TESA6例)进行治疗。结果 改良ICSI在正常受精率(97.5％)、卵裂率(99.05％)、囊胚获得率(28.9％)、临床妊娠率(40.2％)等方面具有显著优势;精子参数与来源不同在ICSI治疗结果上无显著性差异。结论 ICSI是治疗男性不育的有效方法,依据生殖医学理论改良ICSI技术可提高治疗效果,非技术因素也是影响ICSI的重要因素,精子参数与来源不影响ICSI治疗结果。     

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。     

卵母细胞形态与细胞内精子注射后胚胎质量和妊娠率的相关性

细胞内精子注射 (ICSI)是利用在显微外科技术进行附睾精子抽吸、睾丸精子切除、精细胞核周质注射等方法治疗男性不育的主要方式。但成熟因素对配子融合是非常有必要的。为了确定行 ICSI手术时机 ,以及如何选择卵细胞 ,作者做了如下的回顾性研究。方法　选 15 5例利用 ICSI治疗的男性不育患者为研究对象 ,根据胚胎质量好坏分为三组 :A组 6 8例移植了评分为3的胚胎 ,B组 6 0例移植了评分为 3～ 2的胚胎 ,C组 18例移植了评分为 2的胚胎 ,胚胎评分以卵裂速度、有无断裂及卵裂球的大小将质量好的评为 3分 ,质量差的评为 2分。以下列方式刺…     

Cumulative delivery rates after ICSI treatment cycles with freshly retrieved testicular sperm: a 7-year follow-up study

BACKGROUND: The purpose of this study was to assess cumulative delivery rates in patients with non-obstructive or obstructive azoospermia following treatment by testicular sperm extraction (TESE)-ICSI. METHODS: A cohort follow-up study was conducted. Between January 1994 and December 2000, 364 couples with obstructive azoospermia underwent a total of 609 fresh TESE-ICSI treatment cycles. In addition, 303 fresh TESE-ICSI treatment cycles were performed in 235 couples for non-obstructive azoospermia. This study included only patients in whom sperm was recovered. In the non-obstructive group, only patients with maturation arrest, atrophic sclerosis and germ cell aplasia were included. The main outcome measure was a delivery beyond 25 weeks gestation. RESULTS: In patients with obstructive azoospermia, the crude delivery rate after three cycles was 35% while the expected cumulative delivery rate was 48% [95% confidence interval (CI), 41-55]. On the other hand, in patients with non-obstructive azoospermia, the crude cumulative delivery rate after three treatment cycles was 17% while the expected delivery rate was 31% (95% CI, 15-46). A high dropout rate in couples with both non-obstructive and obstructive azoospermia was observed (75 and 50% respectively, after the first cycle). CONCLUSION: This study shows that there is a value in performing several TESE-ICSI attempts in patients with obstructive and non-obstructive azoospermia. The estimates of the non-obstructive group beginning from the third cycle are less reliable due to fewer patients. However, overall, the obstructive group performed better than the non-obstructive group.     

Distribution of spermatogenesis in the testicles of azoospermic men: the presence or absence of spermatids in the testes of men with germinal failure [published erratum appears in Hum Reprod 1998 Mar;13(3):780]

The aim of the study was to determine whether a prior diagnostic testicle biopsy can predict success or failure of testicular sperm extraction (TESE) with intracytoplasmic sperm injection (ICSI) in patients with non-obstructive azoospermia caused by testicular failure, and what is the minimum threshold of sperm production in the testis which must be surpassed for spermatozoa to reach the ejaculate. Forty- five patients with non-obstructive azoospermia caused by testicular failure underwent diagnostic testicle biopsy prior to a planned future TESE-ICSI procedure. The diagnostic testicle biopsy was analysed quantitatively, and correlated with the quantitative findings of spermatogenesis in patients with normal spermatogenesis, as well as with the results of subsequent attempts at TESE-ICSI. Men with non- obstructive azoospermia caused by germinal failure had a mean of 0-6 mature spermatids/seminiferous tubule seen on a diagnostic testicle biopsy, compared to 17-35 mature spermatids/tubule in men with normal spermatogenesis and obstructive azoospermia. These findings were the same for all types of testicular failure whether Sertoli cell only, maturation arrest, cryptorchidism, or post-chemotherapy azoospermia. Twenty-two of 26 men with mature spermatids found in the prior testis biopsy had successful retrieval of spermatozoa for ICSI, 12 of their partners became pregnant, and are either ongoing or delivered. The study suggests that 4-6 mature spermatids/tubule must be present in the testis biopsy for any spermatozoa to reach the ejaculate. More than half of azoospermic patients with germinal failure have minute foci of spermatogenesis which are insufficient to produce spermatozoa in the ejaculate. Prior diagnostic testicle biopsy analysed quantitatively (for the presence of mature spermatids) can predict subsequent success or failure with TESE-ICSI. Incomplete testicular failure may involve a sparse multi-focal distribution of spermatogenesis throughout the entire testicle, rather than a regional distribution. Therefore, it is possible that massive testicular sampling from many different regions of the testes may not be necessary for successful TESE-ICSI.      

Ultrastructural analysis of chromatin defects in testicular spermatids in azoospermic men submitted to TESE-ICSI.

BACKGROUND: Testicular sperm extraction (TESE) combined with intracytoplasmic sperm injection (ICSI) is offered to treat obstructive and non-obstructive azoospermia, but factors that influence the outcome of ICSI are not well defined. METHODS AND RESULTS: The percentage of elongated spermatids with normal chromatin condensation in azoospermic patients submitted for TESE-ICSI was determined. The quantitative analysis could be applied to nine of 19 biopsies classified as incomplete late maturation arrest (LMA) and compared with 10 biopsies with normal spermatogenesis. The percentage of elongated spermatids with normal chromatin was lower in LMA than in normal histology (mean 4.4%, range 0-20, and mean 52.9%, range 40-70 respectively; P = 0.0001). The percentage of elongated spermatids with normal chromatin was negatively correlated with the serum concentration of FSH (r = -0.86, P < 0.0001) and the number of degenerated germ cells per 100 Sertoli cells nuclei (r = -0.68; P < 0.0001), while it was positively correlated with the number of elongating spermatids per 100 Sertoli cell nuclei (r = 0.81; P < 0.0001). The percentage of elongated spermatids with normal chromatin was not correlated with the rate of oocyte fertilization, while the delivery rate/cycle was higher in cases with normal histology compared with cases of LMA. CONCLUSIONS: These preliminary data suggest that an altered chromatin condensation is a ubiquitous defect in spermatids of non-obstructed azoospermic men submitted for TESE-ICSI.     

Y chromosome deletions in azoospermic and severely oligozoospermic men undergoing intracytoplasmic sperm injection after testicular sperm extraction

Y chromosome deletions encompassing the AZFc region have been reported in 13% of azoospermic men and 7% of severely oligozoospermic men. We examined the impact of these Y deletions on the severity of testicular defects in 51 azoospermic men undergoing intracytoplasmic sperm injection (ICSI) after testicular sperm extraction (TESE) and 30 men with severe oligozoospermia undergoing ICSI after ejaculation of spermatozoa. In addition, five azoospermic patients shown previously to have Y chromosome deletions underwent histological evaluation of their previously obtained testis biopsy specimens. A further 27 azoospermic men underwent TESE-ICSI, but not Y chromosome DNA testing. Ten of 51 azoospermic men (20%) who underwent TESE-ICSI and Y-DNA testing were found to be deleted for portions of the Y chromosome AZFc region. Of these 10, five had spermatozoa retrievable from the testis, and in two cases the wives became pregnant. Of the 41 azoospermic men with no Y chromosome deletion, 22 (54%) had spermatozoa retrievable from the testis, and in 12 cases (29%) the wives became pregnant. Four of 30 (13%) severely oligozoospermic patients were found to be deleted for AZFc and in three (75%) of these pregnancy was achieved. The other 26 severely oligozoospermic couples who had no AZFc deletions underwent ICSI, and 12 (46%) have an ongoing or delivered pregnancy. The embryo implantation rate was not significantly different for azoospermic (22%), oligozoospermic (16%), Y-deleted (14%) or Y-intact (18%) men. Of the total of 19 infertile men who had Y chromosome deletions, 14 had deletions within Y chromosome intervals 6D-6F, in the AZFc region. Twelve of those 14 had some spermatozoa (however few in number) in the ejaculate or testis. Five of the Y-deleted men had deletions that extended more proximally on the Y chromosome, and in none of these could any spermatozoa be observed in either ejaculate or testis. These results support the concept that, in azoospermic or oligozoospermic men with Y chromosome deletions limited to intervals 6D-6F (AZFc), there are generally very small numbers of testicular or ejaculated spermatozoa. Larger Y deletions, including and extending beyond the AZFc region and encompassing more Y genes, tend to be associated with a total absence of testicular spermatozoa. In those cases where spermatozoa were retrieved, the presence of Y deletions had no obvious impact on fertilization or pregnancy rate.      

Intraventricular infusions in acute and chronic rabbits

Summary A technique for infusion of chemical substances into the third ventricle of the rabbit is described. A cannula is introduced between the third ventricle and the aqueduct of Sylvius, by means of the stereotaxic method and atlas of Monnier and Gangloff (1961). The effects of the infused substance are tested electrographically with cortical and subcortical recording electrodes. A modification of the stereotaxic socklet enables a permanent implantation of cannula and recording electrodes to be made for experiments in chronic animals.     

Continuous measurement of venous diameter by a combined photoelectric-photographic and plethysmographic technique

Summary The combined photoelectric-photographic and plethysmographic technique allows for the investigation of the distensibility characteristics and pharmacological influence on the capacitance vessels in an uniform vascular bed (skin veins of the isolated rabbit ear). Diameter changes of a vein segment are recorded continuously by means of a photoresistor and are photographed for calibration. Volume changes of the tissue are measured by a water filled plethysmograph and can be compared with diameter changes of the selected vein segment. The method allows for better interpretation of the results obtained by the plethysmographic technique. In particular, it is possible to distinguish between outward filtration and stress-relaxation.This research was supported by Contract, F 44620-71-C-0117 of the USAF School of Aerospace Medicine, European Office of Aerospace Research (OAR) United States Air Force and Deutsche Forschungsgemeinschaft     

纤维树脂桩核修复体力学性能与不同牙体剩余量的关系

背景：牙体组织剩余量与纤维树脂桩核修复体的抗折强度密切相关。 目的：采用体外应力分析方法比较不同牙体残余量对纤维树脂桩核修复体抗疲劳强度的影响。 方法：将36颗离体人下颌单根管前磨牙随机均分为4组,进行牙根管预备与填充后,A组平齐基准平面横断磨除冠部牙体;B组,牙冠近远中斜形损耗,远端牙本质领圈高度2 mm,逐渐减少到近中端时牙本质领圈高度为0 mm;C组,牙冠横行缺失,留2 mm健康牙本质;D组,保留完整冠部牙体组织。各组常规桩道预备后进行黏结处理,将Para post fiber lux 玻璃纤维桩置入桩道内,采用动态力学加载设备对试件进行加载。 结果与结论：4组荷载力值大小为D组> C组> B组> A组,组间两两比较差异有显著性意义(P < 0.05)。实验样本折裂形式大致可分为牙冠斜行折裂、牙颈部横行折裂、牙颈部斜行折裂及牙根折裂等4种。A组发生牙颈部横行折裂7颗、牙颈部斜行折裂2颗;B组发生牙冠斜行折裂4颗、牙颈部斜行折裂5颗;C组发生牙颈部斜行折裂6颗、牙根中部折裂3颗;D组发生牙颈部斜行折裂7颗、牙根中部折裂2颗。表明当牙体残余量较多时,牙齿受到的咬合应力也会从颈缘转移到根部,整体水平也随着减低,纤维树脂桩核修复体的强度也会随着增强,因此在临床实践中应尽可能多保存牙体组织。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

房室交界区三角的观察和测量

在110例人心(成人70,儿童40)上,观察了由冠状窦口、Todaro腱及三尖瓣隔瓣附着缘围成的房室交界区三角,对上述各边界及室间隔膜部进行了测量。三角的三个角各有不同结构占据,前上角为房室结,顶角有冠状窦和心最小静脉开口,后下角深面为右冠状动脉“U”形袢。就上述特点结合临床进行了讨论。