Immunohistochemical observation on luteinizing hormone in rat testes before and after testicular capsulotomy

Aim: In the testicular capsulotomized rats, although there was a significant increase in the luteinizing hormone (LH) levels, the secretion of testosterone remained low. In order to clarify the mechanisms of this phenomenon, the binding of endogenous LH to the testes were observed before and after testicular capsulotomy. Methods: Peroxidase-antiperoxidase (PAP) method was used to detect the binding of LH to the testes in rats. Results: An intense positive staining of LH was found in the Leydig cells of both the normal and sharn-operated control testes. However, at 40 d after operation, the LH immunoreactivity was decreased in the Leydig cells of the capsulotomized testis. By d 60, only very weak positive staining could be observed in these cells. Conclusion: A progressive reduction of endogenous LH binding to the testis occurred in the capsulotomized rat.     

Effect of testicular capsulotomy on fertility of rats

Aim: To demonstrate the effect of capsulotomy on the fertility of male rats. Methods: Testicular capsulotomy was carried out in immature (21 days) and adult (60 days) male rats. The fertility of them was assessed by cohabitation with proestrus females overnight and 20 days later, the females were examined for impregnation. Morphological changes at the site of the capsulotomy were observed under light microscope. Results: In rats capsulotomized at Day 60, the fertility was gradually depressed and all the rats completely lost their fertility 2 months post - operation. At that time, a partial regeneration of the capsule at the site of capsulotomy was observed. Immature rats capsulotomized at Day 21 were found to possess normal fertility at maturity. The capsulotomy site was almost completely recovered 60 days post-operation. Conclusion: In male rats, testicular capsulotomy at the age of Day 60 will damage fertility. However, when capsulotomy is performed at Day 21. fertility is preserve.     

Effect of testicular capsulotomy on lipid droplets in the seminiferous tubules of rats

Aim: In order to reveal the histochemical alteration that might occur during the processes of the spermatogenic disruption induced by testicular capsulotomy, the location and alteration of lipid droplets in the seminiferous tubules were observed in the present study. Methods: Osmium tetroxide was used to demonstrate the lipid droplets in the seminiferous tubules of capsulotomized and sham-operated control testes. Results: In the seminiferous tubules of the sham-operated rat testes, many small lipid droplets were located close to the basement membrane of the seminiferous tubules. But for the capsulotomized testes, the lipid droplets in the seminiferous tubules had increased in size and number, with many lipid droplets migrated towards the lumen of the tubules. Conclusion: The results indicated that a progressive fatty degeneration occurred in the seminiferous tubules after testicular capsulotomy.     

Low sperm recovery from the undescended testis with testicular sperm extraction in postpubertal cryptorchids: preliminary report

As postpubertal cryptorchid testes are generally atrophic and have high malignancy risk, orchiectomy is recommended as the treatment method. However, a few case reports advocated orchiopexy due to finding sperm afterwards. The present clinical study aimed at determining the possibility of finding spermatozoa in postpubertal cryptorchid testis with testicular sperm extraction (TESE). Initially testicular biopsy and later TESE was performed in the undescended testis of 22 unilateral postpubertal cryptorchids and in one of the testes of three bilateral postpubertal cryptorchids. Histopathological assessment of testicular biopsy specimen showed that three patients had maturation arrest and 22 had seminiferous tubular atrophy. By TESE, spermatozoa were found in only one specimen of 25 testes. Our results show that the expectancy to find spermatozoa in postpubertal cryptorchid testis is extremely low by current sperm recovery methods and orchiectomy is still the accurate method of treatment.     

睾丸主要血管或睾丸网旁取精点与取精成功率不具显著相关性：以220例非梗阻性无精子症患者行传统或显微睾丸取精术为例

采用传统与显微睾丸取精术相结合方式对220例梗阻性无精症患者行精子提取术,如果传统手术方式未见精子即为阴性取精位点,然后选取2—3个阴性取精位点行显微睾丸取精术。术中通过手术显微镜评价睾丸血管系统,同时记录阳性取精位点与睾丸血供的关系。总的精子获取率为58-2％,早期应用传统手术方式精子获取率为46．8％,而后行显微睾丸取精术则精子获取率增加11．7％。故显微睾丸取精术的应用可显著增加精子获取率（P=0．017）,且在睾丸网或睾丸主要血管处并不能显著增加阳性取精位点。     

Effect of testicular capsulotomy on secretion of testosterone and gonadotrophins in rats

Aim: In order to clarify further the mechanisms underlying the effect of capsulotomy on testicular function, the levels of testosterone, LH and FSH were observed. Methods: Intratesticular testosterone levels and LH, FSH levels in the peripheral blood of normal, sham-operated and capsulotomized rats were detected by RIA. Results: After testicular capsulotomy, there was a progressive reduction in the testosterone level in the testicular venous blood together with a progressive increase in the LH and FSH levels in the peripheral blood from approximately 30 days post-capsulotomy. Morphological changes were observed at 5-10 days after capsulotomy, i.e., far ahead of the hormonal changes.Conclusion: The seminiferous tubular damage after testicular capsulotomy was not caused by the reduction in testosterone, and on the contrary, the hormonal change might be secondary to the morphological alterations. The increase in LH level most likely resulted from a negative feedback influence from the lowered testosterone level, while the increase in FSH secretion may be a feedback signal of the damaged seminiferous tubules. (Asian J Androl 2000 Dec;2:257-261)     

非梗阻性无精子症患者睾丸体积、生殖激素水平与睾丸穿刺取精结果的相关性研究

目的:探讨非梗阻性无精子症患者睾丸体积、生殖激素水平与睾丸穿刺取精术(TESA)结果的相关性,以及可用于预测TESA结果的睾丸体积、生殖激素水平的切点值,从而为非梗阻性无精子症患者进一步诊疗提供重要资料。方法:121例研究对象均为非梗阻性无精子症患者(NOA),测定其睾丸体积和生殖激素水平,并根据TESA结果分为无精子组和有精子组。结果:无精子组和有精子组的左侧睾丸体积(ml)、右侧睾丸体积(ml)、泌乳素(PRL,ng/ml)、卵泡刺激素(FSH,mIU/ml)、黄体生成素(LH,mIU/ml)、雌二醇(E2,pmol/L)、血清总睾酮(TT,nmol/L)水平分别为7.07±1.06和11.75±1.38、7.37±1.37和11.70±1.98、12.43±11.69和9.60±4.55、15.77±10.84和8.01±7.43、6.12±2.92和8.11±20.11、119.36±43.52和141.12±48.33、11.43±4.05和12.46±4.60。无精子组血清FSH和PRL水平平均值高于有精子组,并且有显著的统计学差异。虽然无精子组的睾丸体积平均数小于有精子组,但两组之间没有统计学差异。对于年龄、血清E2和TT水平,两组之间也没有统计学差异。利用ROC曲线优选的睾丸体积切点值为9 ml,此点其敏感性为93.8%/89.6%(左/右),特异性为100%/94.3%(左/右),睾丸体积ROC曲线的AUC为0.984/0.961(左/右),表明其诊断准确性较高;优选的血清FSH水平切点值为8.18 mIU/ml,此点其敏感性为71.2%,特异性为75.0%,FSH水平ROC曲线的AUC为0.743,表明其诊断准确性中等。结论:睾丸体积和FSH水平对于预测NOA患者TESA结果具有重要意义,并且睾丸体积诊断准确性明显优于FSH。     

Onco‐testicular sperm extraction: testicular sperm extraction in azoospermic and very severely oligozoospermic cancer patients

An increased risk of testicular cancer in men with infertility and poor semen quality has been reported. In view of the high cure rates for testicular germ cell tumours, increasing clinical importance is being placed on the protection of fertility. High‐dose cytostatic therapy may be expected to cause long‐term infertility. Thus, the standard procedure for fertility protection is the cryopreservation of ejaculated spermatozoa or testicular tissue before therapy. Four male patients with azoospermia and two patients with very severe oligozoospermia underwent onco‐testicular sperm extraction (TESE). We attempted onco‐TESE in patients with azoospermia and very severe oligozoospermia after orchiectomy. Of the patients with testicular germ cell tumours, four had spermatozoa in their testicular tissues. Sertoli cell‐only syndrome was found in one patient, and one patient showed maturation arrest without the detection of spermatozoa. Three of six showed seminomatous germ cell tumour, two of six had nonseminomatous germ cell tumour and one patient showed no malignancy. Two patients achieved clinical pregnancy. Fertility challenges in men with cancer are the most straightforward because of the relative ease of obtaining and cryopreserving sperm. Testicular sperm extraction is a useful technique for obtaining spermatozoa before cytotoxic therapy in azoospermic and very severely oligozoospermic cancer patients.     

Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples

Intracytoplasmic sperm injection (ICSI) may be performed with testicular frozen–thawed spermatozoa in patients with nonobstructive azoospermia (NOA). Sperm retrieval can be performed in advance of oocyte aspiration, as it may avoid the possibility of no recovery of spermatozoa on the day of oocyte pickup. There are few studies available in the literature concerning the use of frozen–thawed spermatozoa obtained from testicular sperm aspiration (TESA). To evaluate the effects and the outcomes of ICSI with frozen–thawed spermatozoa obtained by TESA, we performed a retrospective analysis of 43 ICSI cycles using frozen–thawed TESA. We obtained acceptable results with a fertilisation rate of 67.9%, an implantation rate (IR) of 17.1%, and clinical and ongoing pregnancy rates of 41.9% and 37.2% respectively. The results of this study suggest that performing ICSI using cryopreserved frozen–thawed testicular spermatozoa with TESA as a first option is a viable, safe, economic and effective method for patients with NOA.     

Primary testicular plasmacytoma with hydrocele of the testis

A case of primary testicular plasmacytoma complicated with hydrocele of the testis is reported. An 86-year-old man presented with hydrocele of the right testis. High inguinal orchiectomy was performed as the preoperative aspiration cytology of the hydrocele fluid showed atypical cells. Immunohistochemical study of the right testis revealed testicular plasmacytoma positive for IgG. He remained well 9 months after the orchiectomy. This is the second reported case where the preoperative diagnosis of testicular plasmacytoma was made based on the hydrocele fluid cytology.     

Outcome of repeated micro-surgical testicular sperm extraction in patients with non-obstructive azoospermia

Aim： To evaluate the outcome of repetitive micro-surgical testicular sperm extraction （mTESE） attempts in non-obstructive azoospermia （NOA） cases, in relation to patients＇ initial testicular histology results. Methods： A total of 68 patients with NOA in whom mTESE had been performed in previous intracytoplasmic sperm injection （ICSI） attempts were reviewed. Results： Among the 68 patients with NOA, the first mTESE yielded mature sperm for ICSI in 44 （64%） （Sp^＋）, and failed in the remaining 24 （36%） （Sp^-）. Following their first trial, 24 patients decided to undergo a second mTESE. Of these 24 patients, no spermatozoa were obtained in 5 patients, and Sp^＋ but no fertilization/pregnancy were achieved in 19. In these 24 cases, mTESE was successively repeated for two （n = 24）, three （n = 4） and four （n = 1） times. The second attempt yielded mature sperm in 3/5 patients from the Sp group and 16/19 patients from the Sp^＋ group. At the third and fourth trials, 4/4 and 1/1 of the original Sp^＋ patients were Sp^＋ again, respectively. Distribution of main testicular histology included Sertoli cell-only syndrome （16%）, maturation arrest （22%）, hypospermatogenesis （21%） and focal spermatogenesis （41%）. Overall, in repetitive mTESE, 24/29 （82%） of the attempts were finally Sp^＋. Conclusion： Repeated mTESE in patients with NOA is a feasible option, yielding considerably high sperm recovery rate. In patients with NOA, mTESE may safely be repeated one or more times to increase sperm retrieval rate, as well as to increase the chance of retrieving fresh spermatozoa to enable ICSI.     

Influence of motility and vitality in intracytoplasmic sperm injection with ejaculated and testicular sperm

The vitality of spermatozoa used for intracytoplasmic sperm injection (ICSI) is a crucial factor for fertilization, establishment and outcome of a pregnancy in assisted reproductive technique cycles. The sperm origin may also be a limiting factor, although little is known about this issue. It is known that the motility of injected spermatozoa and their origin from ejaculate or testicular biopsies are important predictors in terms of fertilization, pregnancy and birth rates. Oocytes of patients in 2593 cycles were retrieved in our in vitro fertilization programme and inseminated via ICSI. We used motile (group 1, n = 2317) or immotile ejaculated spermatozoa (group 2, n = 79), motile sperm retrieved from testicular biopsies (group 3, n = 62) and immotile spermatozoa from testicular biopsies (group 4, n = 135). Female age and number of oocytes retrieved did not differ significantly among the groups. The fertilization rates were as follows: 67.1% in group 1, 49.8% in group 2, 68.3% in group 3 and 47.8% in group 4. The pregnancy rates in cases where three embryos had been transferred amounted to 35.7% in group 1, 17.3% in group 2, 38.3% in group 3 and 20.5% in group 4. The embryo quality showed no differences between groups 1 and 3 (14.5), and between groups 2 (11.8) and 4 (10.8). The abortion rate was similar in groups 1-3, but increased in group 4 (26.6%, 27.3%, 31.6% and 55.5%). Irrespective of their origin, the fertilization potential of injected spermatozoa was found to be influenced by motility. The resulting pregnancy and birth rates, i.e. the potential of the resulting embryos to implant and to achieve viable pregnancies, seem to be additionally dependent on the sperm origin. This was well shown by declining rates when spermatozoa in a relatively early stage of maturity had been used. We see increasing evidence that the degree of sperm maturity has an important impact on the outcome of ICSI. In obstructive azoospermia, spermatozoa retrieved from the epididymis should be used rather than testicular biopsy spermatozoa, or testicular sperm should be preincubated in culture medium before ICSI.     

Comparison of intracytoplasmic sperm injection outcomes between spermatozoa retrieved from testicular biopsy and from ejaculation in cryptozoospermic men

The infrequent presence of spermatozoa in cryptozoospermic men ejaculate is a limiting factor in the treatment of them. Sometimes, this consideration impels us to apply meticulous microscopic search in ejaculate or testicular sperm extraction (TESE) method. The aim of this study was to assess putative effectiveness of sperm origin, ejaculated or testicular, in cryptozoospermia treatment. In this context, were evaluated intracytoplasmic sperm injection (ICSI) outcomes in two parameters including fertilisation rate (2PN) and embryo quality, independently. We compared the outcome in two groups: patients who underwent ejaculate/ICSI and ones who underwent TESE/ICSI process. Nineteen ICSI cycles performed with testicular spermatozoa and the rest of cycles (n = 208) carried out with ejaculated spermatozoa. Result analysis showed similar fertilisation rate between testicular and ejaculated spermatozoa (respectively, 60% versus 68%, P ≥ 0.05). Also, on the other hand, embryo quality did not show significant differences between two groups, except grade A with low significance. With regard to almost equal performance of both methods in results and being invasive of TESE as surgical sperm retrieval method, the use of ejaculated sperm more than testicular sperm should be recommended in patients with cryptozoospermia whenever possible.     

Combined trifocal and microsurgical testicular sperm extraction is the best technique for testicular sperm retrieval in "low-chance" nonobstructive azoospermia

Background

There is no consensus for the best testicular sperm extraction (TESE) technique in patients with “low-chance” nonobstructive azoospermia (NOA).

Objective

To determine sperm retrieval rates in an intraindividual comparison using three locations of the testicle with and without the assistance of a microscope (microsurgical TESE [M-TESE]).

Design, setting, and participants

A series of 65 patients with low-chance NOA presenting with low testicular volume (<8 ml) and high serum follicle-stimulating hormone (FSH) (>12.4 IU/l) underwent trifocal-TESE plus M-TESE bilaterally (four biopsies per testis).

Intervention

Sperm retrieval was performed as trifocal-TESE (upper, middle, and lower testicular pole) with and without the assistance of a microscope in the middle incision.

Outcome measurements and statistical analysis

The number of evaluated tubules, the mean spermatogenetic scores, and the sperm retrieval rates were evaluated to determine retrieval locations and the use of the microscope. The Friedman and Cochrane Q tests were applied to determine statistical differences. Receiver operating characteristic curves were used for the analysis of serum FSH and testicular volume as preoperative prognostic factors.

Results and limitations

The sperm retrieval success of 66.2% using the combined technique, meaning the percentage of patients with at least one tubule containing elongated spermatids, was the highest in the combination of trifocal- and M-TESE (p < 0.01), indicating this technique as optimal for patients with low-chance NOA. M-TESE and trifocal-TESE alone were not significantly better. The mean spermatogenetic score giving the number of tubules with elongated spermatids in relation to all tubules was significantly higher in M-TESE versus conventional TESE (p < 0.01), indicating the superior quality of the tissue harvested using the microscope. These results are limited by the definition of “success” using “one” spermatid/tubule. Preoperatively, high serum FSH and low testicular volumes did not exclude successful sperm retrieval.

Conclusions

The combination of trifocal- and M-TESE is the best technique to reach high sperm retrieval rates in patients with low-chance NOA.     

Intracytoplasmic sperm injection by testicular sperm in patients with aspermia or azoospermia after cancer treatment

The aim of this retrospective study was to evaluate the efficiency of testicular biopsy and intracytoplasmic sperm injection (ICSI) in patients with aspermia or non-obstructive azoospermia (NOA) after cancer treatment. From 1996 to 2003, 30 men with a history of cancer, affected by aspermia or NOA and without sperm cryopreserved before cytotoxic treatment underwent testicular sperm extraction (TESE). In these men, clinical, hormonal and histological characteristics were compared; 13 underwent 39 TESE-ICSI cycles using frozen-thawed testicular spermatozoa (TESE-ICSI group). In the same period, 31 ICSI cycles were performed in 20 men with aspermia or NOA using ejaculated sperm frozen before cancer treatment (ejaculated sperm-ICSI group). Fertilization, blastocyst development, pregnancy and miscarriage rates were compared between the groups. Testicular volume, serum follicle-stimulating hormone level and Johnsen score indicated complete although reduced spermatogenesis in men with aspermia and abnormal spermatogenesis in men with NOA. After TESE, sperm retrieval was positive in 92% of men with aspermia and 58% of men with NOA. In TESE-ICSI patients with NOA a significantly lower proportion of embryos developed to the blastocyst stage than in patients with aspermia and in those after ICSI with frozen-thawed ejaculated sperm (23% vs. 43% and 47%, p = 0.03 and p < 0.01 respectively). In all groups the miscarriage rates were high; in patients with aspermia and NOA, characterized by increased age, the miscarriage rate tended to be higher in spite of similar female age and female indications of infertility. In patients affected by aspermia or NOA after cancer treatment and without sperm cryopreserved before treatment, TESE-ICSI using testicular sperm provide a chance to father a child.     

Parameters predicting sperm retrieval rates during microscopic testicular sperm extraction in nonobstructive azoospermia

The aim of this study was to investigate the predictive value of the parameters that might have an effect on the success of microscopic testicular sperm extraction (micro‐TESE) in infertile patients with nonobstructive azoospermia (NOA). Between 2003 and 2014, 860 patients with NOA were retrospectively analysed. The effect of age, infertility duration, history of varicocelectomy, herniorrhaphy or orchiopexy, presence of solitary testis, tobacco use, previous testicular biopsy results, history of orchitis, usage of human chorionic gonadotropin in the past three months, presence of undescended or retractile testis, presence of varicocele, testicular volume, levels of serum follicle‐stimulating hormone, luteinising hormone, and testosterone, presence of Klinefelter syndrome and micro‐deletion of Y chromosome on sperm retrieval rates were evaluated. In 45.8% (n = 394) of the patients who underwent micro‐TESE, spermatozoon was adequately obtained. Multiple logistic regression analysis demonstrated that previous successful testicular biopsy (OR = 15.346; GA = 5.45–43.16; p < .001) and higher testicular volumes significantly increase sperm retrieval rate in micro‐TESE. The testicular volume cut‐off as 11 ml was found to be the most significant factor. Although currently testicular biopsy result is not being used as a diagnostic method, it is significantly associated with micro‐TESE result.     

尿毒症患者精子参数变化和睾丸活检病理学研究

目的 探讨男性尿毒症患者精子主要参数的变化和睾丸组织结构病理改变。方法 对8例尿毒症患者的精液检测的同时进行睾丸活检,并进行分析。结果 尿毒症患者的精子活动力(a+b)、存活率和精子密度平均分别为12.2%、30.6%和31.2×106/ml;睾丸活检病变主要表现为生精上皮脱落、管腔阻塞、界膜结缔组织增生、间质水肿等。结论 尿毒症患者精子主要参数(活动力、存活率、密度)均显著降低,睾丸病理改变主要特征为生精功能低下和生精阻滞。     

Open testicular mapping: A less invasive multiple biopsy approach for testicular sperm extraction

This study proposes a testicular sperm extraction technique that was inspired by testicular fine-needle aspiration. Here, we have described the technique of open testicular mapping (OTEM) and evaluated the successful sperm recovery in 92 patients with nonobstructive azoospermia (NOA). All patients underwent an OTEM biopsy. Patients were divided into two groups; group I included men with spermatozoa recovered and group 0 included men without spermatozoa recovered. Age, follicle-stimulating hormone (FSH) level and testicular volume were compared between the groups. In 50 of 92 men (54%), viable spermatozoa were found after OTEM. No differences were noted in age, FSH level or testicular volume. Using OTEM, it was possible to retrieve spermatozoa in 54% of the NOA men.