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.

Microdissection testicular sperm extraction： an update

Patients with non-obstructive azoospermia （NOA） were once considered to be infertile with few treatment options due to the absence of sperm in the ejaculate. In the last two decades, the advent of intracytoplasmic sperm injection （ICSI）, and the application of various testicular sperm retrieval techniques, including fine needle aspiration （FNA）, conventional testicular sperm extraction （TESE） and microdissection testicular sperm extraction （micro-TESE） have revolutionized treatment in this group of men. Because most men with NOA will have isolated regions of spermatogenesis within the testis, studies have illustrated that sperm can be retrieved in most men with NOA, including Klinefelter＇s syndrome （KS）, prior history of chemotherapy and cryptorchidism. Micro-TESE, when compared with conventional TESE has a higher sperm retrieval rate （SRR） with fewer postoperative complications and negative effects on testicular function. In this article, we will compare the efficacy of the different procedures of sperm extraction, discuss the medical treatment and the role of testosterone optimization in men with NOA and describe the micro-TESE surgical technique. Furthermore, we will update our overall experience to allow counseling on the prognosis of sperm retrieval for the specific subsets of NOA.

Seminal plasma anti-Müllerian hormone level correlates with semen parameters but does not predict success of testicular sperm extraction （TESE）

Aim： To assess seminal plasma anti-Müllerian hormone （AMH） level relationships in fertile and infertile males. Methods： Eighty-four male cases were studied and divided into four groups： fertile normozoospermia （n = 16）, oligoastheno- teratozoospermia （n = 15）, obstructive azoospermia （OA） （n = 13） and non-obstructive azoospermia （NOA） （n = 40）. Conventional semen analysis was done for all cases. Testicular biopsy was done with histopathology and fresh tissue examination for testicular sperm extraction （TESE） in NOA cases. NOA group was subdivided according to TESE results into unsuccessful TESE （n = 19） and successful TESE （n = 21）. Seminal plasma AMH was estimated by enzyme linked immunosorbent assay （ELISA） and serum follicular stimulating hormone （FSH） was estimated in NOA cases only by radioimmunoassay （RIA）. Results： Mean seminal AMH was significantly higher in fertile group than in oligoasthenoteratozoospermia with significance （41.5±10.9 pmol/L vs. 30.5±10.3 pmol/L, P 〈 0.05）. Seminal AMH was not detected in any OA patients. Seminal AMH wascorrelated positively with testicular volume （r = 0.329, P = 0.005）, sperm count （r = 0.483, P = 0.007）, sperm motility percent （r = 0.419, P = 0.021） and negatively with sperm abnormal forms percent （r = -0.413, p = 0.023）. Nonsignificant correlation was evident with age （r = -0.155, P = 0.414） and plasma FSH （ r = -0.014, P = 0.943）. In NOA cases, seminal AMH was detectable in 23/40 cases, 14 of them were successful TESE （57.5%） and was undetectable in 17/40 cases, 10 of them were unsuccessful TESE （58.2%）. Conclusion： Seminal plasma AMH is an absolute testicular marker being absent in all OA cases. However, seminal AMH has a poor predictability for successful testicular sperm retrieval in NOA cases.

Testis sperm extraction

The last 20 years have produced developments in the treatment for patients with non-obstructive azoospermia(NOA)who were once considered to be infertile.The combination of intracytoplasmic sperm injection together with various testicular sperm retrieval techniques,including conventional testicular sperm extraction(TESE),microdissection TESE(micro-TESE)and fine needle aspiration(FNA),have revolutionized treatment for these men.In men with NOA,isolated regions of spermatogenesis within the testis are common.The goal for all types of sperm retrieval procedures is locating the focal region(s)of spermatogenesis,and harvesting the sperm for assisted reproduction.This review article explores the surgical management of men with NOA and describes all techniques that can be used for testicular sperm retrieval.A PubMed search was conducted using the key words:“sperm extraction”,“NOA”,“testicular FNA”,“testicular mapping”,“TESE”,and“testicular biopsy”.All articles were reviewed.Articles were included if they provided data on sperm retrieval rates.The methods for performing sperm retrieval rates and outcomes of the various techniques are outlined.Micro-TESE has a higher sperm retrieval rates with fewer postoperative complications and negative effects on testicular function compared with conventional TESE.

Development of a predictive model for increasing sperm retrieval sucpess by microdissection testicular sperm extraction in patients with nonobstructive azoospermia

Microdissection testicular sperm extraction(micro-TESE)is widely used to treat nonobstructive azoospermia.However,a good prediction model is required to anticipate a successful sperm retrieval rate before performing micro-TESE.This retrospective study analyzed the clinical records of 200 nonobstructive azoospermia patients between January 2021 and December 2021.The backward method was used to perform binary logistic regression analysis and identify factors that predicted a successful micro-TESE sperm retrieval.The prediction model was constructed using acquired regression coefficients,and its predictive performance was assessed using the receiver operating characteristic curve.In all,67 patients(sperm retrieval rate:33.5%)underwent successful micro-TESE.Follicle-stimulating hormone,anti-Miillerian hormone,and inhibin B levels varied significantly between patients who underwent successful and unsuccessful micro-TESE.Binary logistic regression analysis yielded the following six predictors:anti-Mullerian hormone(odds ratio[OR]=0.902,95%confidence interval[Cl]:0.821-0.990),inhibin B(OR=1.012,95%Cl:1.001-1.024),Klinefelter’s syndrome(OR=0.022,95%Cl:0.002-0.243),Y chromosome microdeletion(OR=0.050,95%Cl:0.005-0.504),cryptorchidism with orchiopexy(OR=0.085,95%Cl:0.008-0.929),and idiopathic nonobstructive azoospermia(OR=0.031,95%Cl:0.003-0.277).The prediction model had an area under the curve of 0.720(95%Cl:0.645-0.794),sensitivity of 65.7%,specificity of 72.2%,Youden index of 0.379,and cut-off value of 0.305 overall,indicating good predictive value and accuracy.This model can assist clinicians and nonobstructive azoospermia patients in decision-making and avoiding negative micro-TESE results.

An examination of predictive markers for successful sperm extraction procedures:a linear model and systematic review

The authors performed a comprehensive review of current literature to create a model comparing commonly evaluated variables in male factor infertility,for example,follicle-stimulating hormone(FSH),testicular volume(TV),and testosterone(T),to better predict sperm retrieval rate(SRR).Twenty-nine studies were included,9 with data on conventional testicular sperm extraction(cTESE)for a total of 1227 patients and 20 studies including data on microdissection testicular sperm extraction(mTESE)for a total of 4760 patients.A weighted-means value of SRR,FSH,T,and TV was created,and a weighted linear regression was then used to describe associations among SRR,type of procedure,FSH,T,and TV.In this study,weighted-means values demonstrated mTESE to be superior to cTESE with an SRR of 51.9%vs 40.1%.Multiple weighted linear regressions were created to describe associations among SRR,procedure type,FSH,T,and TV.The models showed that for every 1.19 mIU ml^(−1)increase in FSH,there would be a significant decrease in SRR by 1.0%.Seeking to create a more clinically relevant model,FSH values were then divided into normal,moderate elevation,and significant elevation categories(FSH<10 mIU ml^(−1),10–19 mIU ml^(−1),and>20 mIU ml^(−1),respectively).For an index patient undergoing cTESE,the retrieval rates would be 57.1%,44.3%,and 31.2%for values normal,moderately elevated,and significantly elevated,respectively.In conclusion,in a large meta-analysis,mTESE was shown to be more successful than cTESE for sperm retrievals.FSH has an inverse relationship to SRR in retrieval techniques and can alone be predictive of cTESE SRR.

Testicular sperm extraction(TESE)outcomes in the context of malignant disease:a systematic review

Advances in the oncology field have led to improved survival rates.Consequently,quality of life after remission is anticipated,which includes the possibility to conceive children.Since cancer treatments are potentially gonadotoxic,fertility preservation must be proposed.Male fertility preservation is mainly based on ejaculated sperm cryopreservation.When this is not possible,testicular sperm extraction(TESE)may be planned.To identify situations in which TESE has been beneficial,a systematic review was conducted.The search was carried out on the PubMed,Scopus,Google Scholar,and CISMeF databases from 1 January 2000 to 19 March 2020.Preferred Reporting Items for Systematic Reviews and Meta-Analyses(PRISMA)recommendations were followed in selecting items of interest.Thirty-four articles were included in the systematic review,including 15 articles on oncological testicular sperm extraction(oncoTESE),18 articles on postgonadotoxic treatment TESE and 1 article on both oncoTESE and postgonadotoxic treatment TESE.Testicular sperm freezing was possible for 42.9%to 57.7%of patients before gonadotoxic treatment and for 32.4%to 75.5%of patients after gonadotoxic treatment,depending on the type of malignant disease.Although no formal conclusion could be drawn about the chances to obtain sperm in specific situations,our results suggest that TESE can be proposed before and after gonadotoxic treatment.Before treatment,TESE is more often proposed for men with testicular cancer presenting with azoospermia since TESE can be performed simultaneously with tumor removal or orchiectomy.After chemotherapy,TESE may be planned if the patient presents with persistent azoospermia.

Combination of serum inhibin B and follicle-stimulating hormone levels can not improve the diagnostic accuracy on testicular sperm extraction outcomes in Chinese non-obstructive azoospermic men

Background It is still controversial whether the serum inhibin B level is a superior predictor of the presence of sperm in testicular sperm extraction （TESE） in azoospermic men compared with serum follicle-stimulating hormone （FSH）. In this study, we evaluated the diagnostic accuracy of serum inhibin B levels as a predictor of the outcome of TESE in Chinese non-obstructive azoospermic men and compared it with the traditional marker serum FSH and testicular volumes. Methods Basal values of serum hormone levels, testicular volumes and histological evaluation of 305 Chinese non-obstructive azoospermic men were analyzed. The level of inhibin B was measured using a three-step enzyme-linked immunoassay before sperm extraction, and the diagnostic accuracy of prediction of the outcome of TESE was compared for different markers by the receiver operating characteristics （ROC） curve analysis. Results Testicular sperm was successfully retrieved in 137 of 305 patients （44.9%）. The serum level of inhibin B, the FSH and the testicular volume were significantly different between the successful TESE group and the unsuccessful group. According to the ROC curve analysis, for inhibin B, the cut-off value for discriminating between successful and failed TESE was 28.39 pg/ml （sensitivity 83.5%, specificity 79.1%）. For FSH, the best cut-off value for discriminating was 11.05 pg/ml （sensitivity 83.5%, specificity 74.5%）. The area under the ROC curve of serum inhibin B was similar to that of FSH. Combining the serum inhibin B with FSH levels did not improve the predictive value for successful TESE. Conclusions Serum inhibin B and FSH levels are correlated with spermatogenesis. However, inhibin B is not superior to FSH in predicting the presence of sperm in TESE. And the combination of them does not improve the diagnostic accuracy on TESE outcome.

Sperm retrieval rates and clinical outcomes for patients with different causes of azoospermia who undergo microdissection testicular sperm extraction-intracytoplasmic sperm injection

The aim of our study was to compare the sperm retrieval rates(SRRs)and clinical outcomes of patients with different causes of azoospermia who underwent microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI).We conducted a retrospective study at the Reproductive Medicine Center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.This study examined 769 patients with nonobstructive azoospermia who underwent 347 cycles of micro-TESE-ICSI.Patients with azoospermia were classified into Group A(Klinefelter syndrome,n=284,125 cycles),Group B(azoospermia Y chromosome factor c[AZFc]microdeletion,n=91,64 cycles),Group C(cryptorchidism,n=52,39 cycles),Group D(previous mumps and bilateral orchitis,n=23,23 cycles),and Group E(idiopathic azoospermia,n=319,96 cycles).Clinical characteristics,SRR,embryonic development,and pregnancy outcomes of the patients were compared between all groups.Patients in Group D had the highest and most successful SRR.The average SRR for all patients was 46.0%.The rates of clinical pregnancy,implantation,and live birth in Group D were 78.3%,65.0%,and 74.0%,respectively,which were higher than those in all other groups(P<0.05).Group B patients had the lowest clinical pregnancy,implantation,and live birth rates of all groups(P<0.05).No differences were found in the miscarriage rate or birth defects among the groups(P>0.05).Patients with orchitis had the highest SRR and best clinical outcomes.Although AZFc microdeletion patients had a higher SRR,their clinical outcomes were worse.

Testicular sperm extraction and intracytoplasmic sperm injection in non obstructive azoospermia

Objective To report the experience in sperm extraction from testicular biopsies (TESE) performed from March 1996 to July 1998 in men with non obstructive azoospermia (NOA) Methods Comparisons of age, volume of both testes, serum FSH and testosterone in men, and histology of testicular samples in the first cycles between cycles with spermatozoa found and those without spermatozoa found were performed Comparisons of fertilization, cleavage and pregnancy rates between cycles with spermatozoa injected and those with spermatids injected were performed Results Spermatozoa were found in only 12 out of 26 first TESE cycles (46 2%) and other cycles had spermatids (round cells) only Age of men, history of mumps orchitis/oligozoospermia, volume of both testes and serum FSH/testosterone levels in men were not significantly different between cycles with and without spermatozoa The fertilization rate was significantly higher in cycles with spermatozoa injected than those with round cell injections (63 3% vs 23 2%, P <0 0001, Chi squared test) The pregnancy rate was 14 3% per cycle when spermatozoa were injected Conclusions TESE followed by Intracytoplasmic sperm injection (ICSI) is an effective treatment in patients with NOA Less than half of the patients undergoing TESE had spermatozoa recovered Age of men, volume of both testes and serum FSH/ testosterone levels in men were not useful in predicting successful recovery Compared to using ejaculated and epididymal spermatozoa, fertilization and pregnancy rates were achieved when testicular spermatozoa were used for ICSI

Clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection with fresh or cryopreserved sperm in patients with nonobstructive azoospermia

We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI)treatment that used fresh or cryopreserved sperm in patients with nonobstructive azoospermia(NOA).A total of 338 NOA patients with 344 consecutive cycles received treatment in the reproductive medicine center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.Fresh oocytes and fresh sperm were used in 222 patients with 234 cycles(Group A).Fresh oocytes and cryopreserved sperm were used in 116 patients with 110 cycles(Group B).We compared patient characteristics,embryonic development,and pregnancy outcomes between Groups A and B.There was no statistical difference in the patient characteristics,and no differences were observed with fertilization or quality embryo rates between Groups A and B.The rates of clinical pregnancy and live birth were both higher for Group A than those for Group B(both P<0.05).In conclusion,fresh testicular sperm appears to produce better ICSI outcomes than cryopreserved testicular sperm in patients with NOA.

Predictive factors for successful sperm retrieval by microdissection testicular sperm extraction in men with nonobstructive azoospermia and a history of cryptorchidism

This study aims to explore the factors influencing the success rate of the microdissection testicular sperm extraction(Micro-TESE)in patients with nonobstructive azoospermia(NOA)and cryptorchidism.Clinical data of 162 patients with cryptorchidism who underwent Micro-TESE due to infertility from December 2015 to May 2020 in the First Affiliated Hospital of Nanjing Medical University were analyzed retrospectively.In the univariate analysis,significant differences in the age of patient at the time of orchidopexy(median[interquartile range,IQR]:7.0[4.0–11.0]years vs 11.5[9.0–14.5]years,P<0.001),interval between orchidopexy and Micro-TESE(mean±standard deviation:17.5±5.0 years vs 14.4±4.4 years,P<0.001),severity of cryptorchidism(unilateral[62.8%]vs bilateral[31.6%],P<0.001;location of cryptorchidism,intra-abdominal[27.3%]vs inguinal[44.8%]vs suprascrotal[66.7%],P<0.001),volume of the dominant testis(median[IQR]:17.00[15.00–19.00]ml vs 14.50[11.75–16.25]ml,P<0.001),and levels of follicle-stimulating hormone(FSH;P=0.004)and testosterone(P=0.006)were observed between the successful and failed sperm extraction groups.After conducting the multivariate analysis,four of these factors,including unilateral/bilateral cryptorchidism(P<0.001),location of cryptorchidism(P=0.032),age of orchidopexy(P<0.001),and dominant testicular volume,were adopted in the clinical prediction model to evaluate preoperatively the success rate of Micro-TESE for patients with NOA and cryptorchidism.The likelihood of successful sperm retrieval by Micro-TESE in men with NOA and cryptorchidism increased in patients with mild forms of cryptorchidism.

A preliminary study of sperm identification in microdissection testicular sperm extraction samples with deep convolutional neural networks

Sperm identification and selection is an essential task when processing human testicular samples for in vitro fertilization.Locating and identifying sperm cell(s)in human testicular biopsy samples is labor intensive and time consuming.We developed a new computer-aided sperm analysis(CASA)system,which utilizes deep learning for near human-level performance on testicular sperm extraction(TESE),trained on a custom dataset.The system automates the identification of sperm in testicular biopsy samples.A dataset of 702 de-identified images from testicular biopsy samples of 30 patients was collected.Each image was normalized and passed through glare filters and diffraction correction.The data were split 80%,10%,and 10%into training,validation,and test sets,respectively.Then,a deep object detection network,composed of a feature extraction network and object detection network,was trained on this dataset.The model was benchmarked against embryologists’performance on the detection task.Our deep learning CASA system achieved a mean average precision(mAP)of 0.741,with an average recall(AR)of 0.376 on our dataset.Our proposed method can work in real time;its speed is effectively limited only by the imaging speed of the microscope.Our results indicate that deep learning-based technologies can improve the efficiency of finding sperm in testicular biopsy samples.

Laboratory and Clinical Outcomes of Single Sperm Cryopreservation in Patients Underwent Micro-TESE

For men with severe oligozoospermia, sperm cryopreservation can preserve surgically obtained sperm. How to cryopreserve single sperm in men is still a hot topic in assisted reproduction technology. Aim to analyze the laboratory and pregnancy outcomes of single sperm cryopreservation group, we retrospectively selected 38 cycles underwent single sperm cryopreservation and thawing as the study group and 618 cycles underwent conventional sperm cryopreservation and thawing as the control group, which were performed in the reproductive medicine center of the Sixth Affiliated Hospital, Sun Yatsen University, from April 2014 to October 2023. All the sperm came from microdissection testicular sperm extraction (micro-TESE), and performed intracytoplasmic sperm injection (ICSI) for fertilization. Zygotes were cultured to Day 3 embryo, which were freshly transferred to female uterus. Surplus embryos were cultured to blastosphere and cryopreserved. There was no statistical difference in female/male age, female BMI, infertility duration and female basal sex hormone (FSH, LH E2, AMH), No. of oocytes retrieved per cycle, No. of ICSI oocytes per cycle and No. of embryos transferred per cycle between the two groups (P > 0.05). No significant difference was found in two-pronuclear oocyte fertilization rate (59.23% VS 58.84%), Day 3 available embryo rate (61.81% VS 63.55%), Day 3 good-quality embryo rate (45.73% VS 50.27%), blastocyst formation rate (47.83% VS 49.46%), the implantation rate (47.37% VS 52.16%), clinical pregnancy rate (36.84% VS 47.18%), miscarriage rate (14.29% VS 12.68%) and live birth rate (85.71% VS 81.70%) between two groups (P > 0.05). In conclusion, single-sperm cryopreservation was the optimal method to preserve sperm after micro-TESE. It can increase the utilization of each sperm and lead to clinical pregnancy.

No relationship between biopsy sites near the main testicular vessels or rete testis and successful sperm retrieval using conventional or microdissection biopsies in 220 non-obstructive azoospermic men

In 220 consecutive patients with non-obstructive azoospermia, sperm retrieval was attempted by a combination of conventional and microdissection testicular sperm extraction （TESE）. For sperm retrieval, 2-3 conventional biopsies were performed followed by a microdissection TESE in cases of negative conventional biopsies. During the surgery, the vasculature of the testis was assessed using the operative microscope, and the location of positive biopsies was registered in relation to the blood supply. The overall sperm retrieval rate was 58.2%. From the initial conventional biopsies, sperm could be retrieved in 46.8% of the patients. With microdissection TESE, sperm could be retrieved from an additional 11.4% of the patients. The further use of microdissection TESE improved the sperm retrieval rate significantly （P=0.017）. No significant accumulation of positive biopsies was found towards the rete testis or the main testicular vessels.

Probability to retrieve testicular spermatozoa in azoospermic patients

Aim: The degree of probability to retrieve spermatozoa from testicular tissue for intracytoplasmic sperm injection intooocytes is of interest for counselling of infertility patients. We investigated the relation of sperm retrieval to clinical dataand histological pattern in testicular biopsies from azoospermic patients. Methods: In 264 testicular biopsies from142 azoospermic patients, the testicular tissue was shredded to separate the spermatozoa, histological semi-thin sec-tions of which were then evaluated using Johnsen score. Results: The retrieval of spermatozoa correlated signifi-cantly (P < 0.001) with the testicular volume (r = 0.49), the FSH concentration ( r = -0.66), the maximum score(r = 0.85) and the mean Johnsen score (r = 0.81). In the multivariate regression analysis the successful testicularsperm extraction showed the closest relationship to the maximum score. The testicular volume correlated significantlywith the mean Johnsen score ( r = 0.64, P < 0. 001), and the basal serum FSH concentration mainly with the maxi-mum score ( r = -0.77; P < 0. 001 ). Patients with a history cryptorchidism showed a significantly lower Johnsenscore compared to the patients who did not have any testicular disease in the past (3.7 ± 2.4 vs. 5.9 ± 2. 5; P < O.01). Conclusion: In a limited range, the testicular volume and the FSH concentration in serum were related to theJohnsen score which correlated significantly with the sperm retrieval. The successful sperm retrieval can be expected inall azoospermic patients irrespective of the results of clinical examination. However, the probability of retrieval of sper-matozoa decreased significantly in patients with a FSH level > 18 U/L, tesficular volume < 5mL, mean Johnsen score< 5, and maximum Johnsen score < 7.

A laboratory modification to testicular sperm preparation technique improves spermatogenic cell yield

Testicular sperm extraction is a common procedure used to find spermatogenic cells in men with nonobstructive azoospermia. The laboratory processing of biopsied testicular tissues needs to be performed meticulously to acquire a high yield of cells. In this study, the effectiveness of mincing the tissues after testicular biopsy was assessed using histological evaluation, as was the possible adverse effect of residual tissue on the migration of spermatogenic cells during density gradient centrifugation. Our results indicate that testicular residual tissue, when laid on the density gradient medium along with the sperm wash, hinders the spermatogenic cells＇ forming a pellet during centrifugation, and therefore impairs the intracytoplasmic sperm injection procedure. Whereas the mean number of recovered cells from the sperm wash medium （SWM） with residual tissue is 39.435 ~ 24.849, it was notably higher （60.189 ~ 28.214 cells） in the SWM without minced tissues. The remaining tissue contained no functional seminiferous tubules or spermatogenic cells in histological sections. In conclusion, the remaining residual tissue after mincing biopsied testicular tissue does not add any functional or cellular contribution to spermatogenic cell retrieval; in fact, it may block the cellular elements in the accompanying cell suspension from migrating through the gradient layers to form a pellet during centrifugation and cause loss of spermatogenic cells.

Aberrant methylation of the TDMR of the GTF2AIL promoter does not affect fertilisation rates via TESE in patients with hypospermatogenesis

Increasing evidence shows a relationship between epigenetic regulation and male infertility. The GTF2A1L gene promoter contains the DNA methylation site of a tissue-specific differentially methylated region （TDMR）. Eighty-six patients with non-obstructive azoospermia were assessed for the DNA methylation state of CpG islands in the GTF2AIL promoter using testicular genomic DNA. Based on histological criteria, 26 of the 86 patients had normal spermatogenesis （controls）, 17 had hypospermatogenesis and 26 had a Sertoli cell-only phenotype or tubular sclerosis. GTF2AIL TDMR methylation was significantly lower in testes DNA from control samples than from hypospermatogenic samples （P=0.029）. Patients with hypospermatogenesis were divided into two subgroups： high DNA methylation （HM, n=5） and low DNA methylation （LM, n= 12）. The GTF2AIL TDMR methylation rate differed significantly between the HM and LM groups （P=0.0019）, and GTF2A 1L expression was significantly higher among the LM than in the HM patients （P=0.023）. High TDMR methylation was correlated with low GTF2AIL gene expression levels. Both groups demonstrated relatively good outcomes with respect to sperm retrieval, fertilisation, pregnancy and childbirth rates. We observed that aberrant GTF2AIL gene expression was not correlated with fertilisation rates. The testicular sperm extraction （TESE） technique may be used to overcome male infertility due to aberrant TDMR methvlation.

Testicular volume is a noninvasive predictor of sperm retrieval failure in idiopathic nonobstructive azoospermia

We investigated the prognostic importance of noninvasive factors in predicting sperm retrieval failure in idiopathic nonobstructive azoospermia(iNOA).We studied 193 patients with nonobstructive azoospermia who underwent microsurgical testicular sperm extraction.The Chi-square test and Mann–Whitney U tests for clinical parameters and seminiferous tubule distribution were used for between-group comparisons.A logistic regression analysis was conducted to identify predictors of retrieval failure.Area under the receiver operating characteristic curve for each variable was evaluated,and the net clinical benefit was calculated using a clinical decision curve.Patients with iNOA had a lower sperm retrieval rate than those with known causes.Moreover,testicular volume was an independent factor affecting sperm extraction outcomes(odds ratio=0.79,P<0.05).The testicular volume cut-off value was 6.5 ml(area under the curve:0.694).The patients with iNOA were categorized into two groups on the basis of the distribution of seminiferous tubules observed.The sperm retrieval rate and testicular volume were significantly different between the groups with a uniform or heterogeneous tubule distribution.There was also a significant association between a uniform tubule distribution and testicular volume.In conclusion,a testicular volume of more than 6.5 ml effectively predicts microsurgical testicular sperm extraction failure due to a uniform tubule distribution in patients with iNOA.

Machine learning-based prediction of pregnancy outcomes in couples with non-obstructive azoospermia using micro-TESE for ICSI: a retrospective cohort study

Objective:To develop a clinically applicable tool for predicting clinical pregnancy,providing individualized patient counseling,and helping couples with non-obstructive azoospermia(NOA)decide whether to use fresh or cryopreserved spermatozoa for oocyte insemination before microdissection testicular sperm extraction(mTESE).Methods:A total of 240 couples with NOA who underwent mTESE-ICSI were divided into two groups based on the type of spermatozoa used for intracytoplasmic sperm injection(ICSI):the fresh and cryopreserved groups.After evaluating several machine learning algorithms,logistic regression was selected.Using LASSO regression and 10-fold cross-validation,the factors associated with clinical pregnancy were analyzed.Results:The area under the curves(AUCs)for the fresh and cryopreserved groups in the Logistic Regression-based prediction model were 0.977 and 0.759,respectively.Compared with various modeling algorithms,Logistic Regression outperformed machine learning in both groups,with an AUC of 0.945 for the fresh group and 0.788 for the cryopreserved group.Conclusion:The model accurately predicted clinical pregnancies in NOA couples.