Case report: remedial microdissection testicular sperm extraction after onco-microdissection testicular sperm extraction failure.

BACKGROUND: Testicular cancer (TC) mostly occurs in men aged 14 to 44. Studies have shown that TC seriously damages male fertility, and 6% to 24% of patients with TC were even found to suffer from azoospermia when they are diagnosed. At present, some studies have pointed out that onco-microdissection testicular sperm extraction (mTESE) can extract sperm from tumor testicles. However, there are almost no reports on remedial measures after onco-mTESE failure. Given the valuable opportunity for fertility preservation in patients with TC and azoospermia, it is necessary to provide effective remedial methods for patients with failed onco-mTESE. METHODS: Two young men, who were diagnosed with TC and also found to have azoospermia, tried onco-mTESE while undergoing radical orchiectomy for fertility preservation. However, sperm extraction failed in both patients. Subsequently, the isolated testicular tissue of the patient in case 1 suffered from TC again, and the patient in case 2 was scheduled to receive multiple cycles of gonadotoxic chemotherapy. Because both had a plan to have a birth in the future, we performed remedial mTESE. RESULTS: Sperm was successfully extracted from both patients. The patient recovered well, without complications. The patient couple in case 1 underwent 1 intracytoplasmic sperm injection (ICSI) cycle but did not achieve clinical pregnancy. CONCLUSIONS: There is still an opportunity to extract sperm successfully using onco-mTESE, despite the difficulty of fertility preservation in TC patients with azoospermia. If sperm extraction from the tumor testis fails, implementing remedial mTESE as early as possible would likely preserve the last chance of fertility for these patients.

A Prediction Model of Sperm Retrieval in Males with Idiopathic Non-obstructive Azoospermia for Microdissection Testicular Sperm Extraction.

Patients with Idiopathic non-obstructive azoospermia (iNOA) can achieve fertility by extracting testicular sperm through microdissection testicular sperm extraction (mTESE). But more than half of iNOA patients still cannot benefit from mTESE. In recent years, some studies had reported that serum hormones may be related to the outcome of sperm retrieval, but few had been verified. We hope to obtain a predictive method that is convenient for clinical application and can help judge the outcome of sperm extraction before implementing mTESE. We performed a retrospective analysis of NOA patients who underwent mTESE in the same andrology center from June 2020 to November 2022. A total of 261 patients with complete data were collected, logistic regression analysis was performed and a predictive model was constructed. Then, from December 2022 to May 2023, one prospective cohort of 48 NOA patients who met the inclusion criteria from the same center was recruited to validate the risk prediction model. We successfully constructed a logistic regression model to predict the outcome of iNOA patients undergoing mTESE and found that higher serum anti-Müllerian hormone (AMH) levels were associated with failure sperm retrieval, resulting in an AMH cut-off of 2.60 ng/ml. The area under the receiver operating curve was 0.811, the sensitivity was 0.870, and the specificity was 0.705. Decision curve analysis demonstrated that the threshold probability was above 4%, and unnecessary mTESE could be reduced using this model. In a prospective cohort at the same center, 85.42% (41/48) of iNOA patients correctly identified the mTESE outcome using this model. A logistic regression model with AMH as an independent predictor can predict mTESE outcomes in iNOA patients. Preoperative selection of mTESE in patients with iNOA using this model had clinical benefit in reducing unnecessary surgery. The model demonstrated good accuracy in a small prospective cohort validation.

Complexation of Am(III) and Eu(III) with DRAPA (N,N-dialkyl-6-amide-pyridine-2-carboxylic acid).

The similarity of the coordination chemistry of Am(III) and Eu(III) and two homologous tridentate ligands, N,N-di-2-ethylhexyl-6-amide-pyridine-2-carboxylic acid (DEHAPA, HL') in solvent extraction and N,N-dimethyl-6-amide-pyridine-2-carboxylic acid (DMAPA, HL) in aqueous solution and in the solid state, is revealed structurally and spectroscopically with complexes ML'3 (org), ML3 (aq) and ML3 (s), respectively.

Identification of biallelic variations of CEP70 in patients with male infertility.

Introduction: Male infertility is a severe health issue caused by complex and multifactorial pathological conditions. Genetic factors are a major cause of male infertility. CEP70, a centrosomal protein, has been reported to play an important role in male reproduction in mice. However, the role of CEP70 in human male infertility is limited. Methods: Whole exome sequencing and Sanger sequencing were used to identify the genetic cause of the infertile patients. Papanicolaou staining, scanning electron microscopy and transmission electron microscopy were further conducted to explore morphological and ultrastructural defects in spermatozoa from the patient. Immunofluorescence staining was used to detect the pathogenicity of the identified variants and the particular expression of CEP70 in testis. Results: In this study, we identified biallelic mutations of CEP70 in two unrelated infertile male individuals with oligoasthenoteratozoospermia that followed a recessive inheritance pattern. Papanicolaou staining, scanning electron microscopy and transmission electron microscopy showed that morphological and ultrastructural defects in the acrosome and flagellum of sperm from the patient in a pattern strikingly similar to that in Cep70-/- male mice. The results of immunofluorescence staining suggested that CEP70 was normally expressed in the acrosome and flagellum of control sperm but was hardly detected in the sperm of patient carrying CEP70 variation. We also explored the particular expression pattern of CEP70 during spermatogenesis in humans and mice. Conclusions: Biallelic mutations of CEP70 might be a novel genetic cause of human male infertility, which could potentially serve as a basis for genetic counseling and diagnosis of male infertility.

Outcomes comparison of testicular versus ejaculated sperm for intracytoplasmic sperm injection in infertile men with high DNA fragmentation: updated systematic review and meta-analysis.

Background: The testicular sperm instead of ejaculated sperm for intracytoplasmic sperm injection (ICSI) in infertile men with high sperm DNA fragmentation (SDF) is a controversial topic. This updated systematic review and meta-analysis aims to evaluate whether couples with high level of SDF will benefit more from intracytoplasmic sperm injection with testicular sperm (Testi-ICSI) as compared to intracytoplasmic sperm injection with ejaculated sperm (Ejac-ICSI). Methods: A systematic search was conducted according to PRISMA guidelines, using PubMed, Embase, Web of Science and the Cochrane Central Register of Controlled Trials (CENTRAL), encompassing studies from the earliest record until May 2022. We included studies analyzing comparative pregnancy outcomes of testicular versus ejaculated sperm for ICSI in infertile men with high DNA fragmentation. The risks of bias and certainty of evidence were assessed using the Risk Of Bias In Non-randomized Studies of Interventions (ROBINS-I) and the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) framework, respectively. Results: Eleven studies were included. Meta-analysis showed that SDF levels revealed a significant difference association [odds ratio (OR) =-25.81; 95% confidence interval (CI): -34.82, -16.81; I2=94%; P<0.00001] between testicular and ejaculated sperm. Compared with Ejac-ICSI, a non-significant tendency was observed for fertilization rates (FRs) in the Testi-ICSI group (OR =0.87; 95% CI: 0.67, 1.12; I2=81%; P=0.28). However, there was significant difference pointing to better outcomes for Testi-ICSI in clinical pregnancy rates (CPRs) (OR =2.36; 95% CI: 1.71, 3.24; I2=0%; P<0.00001), live birth rates (LBRs) (OR =3.10; 95% CI: 2.13, 4.51; I2=4%; P<0.00001) and miscarriage rates (MRs) (OR =0.28; 95% CI: 0.13, 0.60; I2=0%; P=0.001). Conclusions: Results of this updated meta-analysis reveal that SDF rates are lower in testicular sperm than in ejaculated sperm and that Testi-ICSI is correlated with better clinical outcomes, including higher CPRs, higher LBRs, and lower MRs in infertile males with high SDF levels. Nevertheless, with the overall low to moderate quality of the studies, further well-designed controlled studies are required.

Case report: Biallelic variants in POLR3B gene lead to 4H leukodystrophy from the study of brother and sister.

INTRODUCTION: 4H leukodystrophy, one of POLR3-related leukodystrophy, is a rare hereditary brain white matter disease caused by the pathogenic biallelic variations in POLR3A, POLR3B, or POLR1C. Hypomyelination, hypodontia, and hypogonadotropic hypogonadism is mainly presented in patients with 4H leukodystrophy. PATIENT CONCERNS: Here, we reported the brother and the sister with new compound heterozygous (c.1615G>T and c.165-167del) with various degrees of phenotypes including dysbasia, myopia, dental abnormal, and hypogonadotropic hypogonadism. DIAGNOSIS: The brother and sister were diagnosed with 4H leukodystrophy. INTERVENTIONS: Gonadotrophins treatment of the brother could significantly improve the development of secondary sexual characteristics and genitalia. OUTCOMES: This study showed that the same genotype of POLR3B may have variable clinical phenotypes in the brother and sister. CONCLUSION: The exploration of molecular functions and genetic counseling are crucial for further diagnosis and treatment of POLR3-related leukodystrophy.

A novel splice site variant in ANOS1 gene leads to Kallmann syndrome in three siblings.

Idiopathic hypogonadotropic hypogonadism (IHH) is a rare genetic disease caused by low doses of hypothalamic gonadotropin-releasing hormone (GnRH), leading to absence or delayed sexual development. Kallmann syndrome (KS) is characterized by IHH with anosmia or hyposmia. Here, we identified a novel splice site variant (c. 726+2T>G) of ANOS1 gene in three siblings with KS from a Chinese Han family by whole-exome sequencing (WES). In this family, KS is classified as an X-linked recessive inheritance pattern. This mutation was inherited from the mother by Sanger sequencing. An in vitro functional experiment has identified the deleterious effect of this mutation on the transcriptional level of ANOS1 gene. Importantly, the effectiveness of timely hormone replacement therapy was evaluated on the three siblings. Hence, finding genetic causes could be helpful in the early diagnosis and timely treatment of KS.

A homozygous RNF220 mutation leads to male infertility with small-headed sperm.

Morphological abnormality of spermatozoa head is a human infertility syndrome caused by spermatogenesis defects. The genetic alterations associated with macrozoospermia, globozoospermia and acephalic spermatozoa are relatively definite, whereas the underlying pathogenesis of small-headed sperm remains unclear. Here, we report a 32-year-old infertile male from a consanguineous family, who presented with 95% of small-headed sperm. Subsequent whole-exome sequencing (WES) analysis identified a homozygous mutation (c. 56C>T [p. P19L]) in Ring Finger Protein 220 (RNF220) gene, which is dominantly expressed in testis. RNF220 protein is an E3 ligase promoting the ubiquitination and proteasomal degradation of SIN3 Transcription Regulator Family Member B (SIN3B), a chromatin-associated protein, which is primarily expressed in the nucleus of germ cells and plays a vital role in chromatin condensation. Notably, this variant could promote the RNF220 protein degradation leading to excessive condensed chromatin in the sperm through the medium SIN3B. Therefore, our study is the first to identify an autosomal recessive genetic mutation in RNF220 that was responsible for small-headed sperm in humans by regulating the expression of chromatin-associated protein SIN3B.

A risk prediction model of sperm retrieval failure with fine needle aspiration in males with non-obstructive azoospermia.

STUDY QUESTION: Can we predict the risk of sperm retrieval failure among men with non-obstructive azoospermia (NOA) before they undergo fine needle aspiration (FNA)? SUMMARY ANSWER: Our model, which includes FSH level, age and testicular volume as variables, can predict the risk of sperm retrieval failure with FNA. WHAT IS KNOWN ALREADY: Combined with ICSI, testicular sperm aspiration (TESA) can enable patients with NOA to have their own genetic offspring. Nearly all reproductive medicine centres in China have applied FNA, but approximately half of patients with NOA experience testicular sperm retrieval failure. Nevertheless, the models developed to predict the likelihood of obtaining spermatozoa with testicular sperm extraction (TESE) cannot accurately predict sperm retrieval, and few of these models have been sufficiently validated. STUDY DESIGN, SIZE, DURATION: This study involved three cohorts including 597 men with NOA. From 1 January 2015 to 31 July 2017, a retrospective cohort of 317 males with NOA who underwent FNA procedures at a university affiliated hospital were included to build a risk prediction model of sperm retrieval failure with FNA. Then, from 25 October 2017 to 31 March 2018, two prospective cohorts of 61 and 219 males with NOA from the same hospital and one other reproductive specialist hospital respectively, were recruited to validate the risk prediction model. PARTICIPANTS/MATERIALS, SETTING, METHODS: All men with NOA undergoing their first TESE procedure as part of a fertility treatment were included. The primary end-point was the presence of one or more spermatozoa (regardless of their motility) obtained with FNA. A binary multivariable logistic model was built to predict the risk of sperm retrieval failure after TESA using the dataset from the retrospective cohort. A cut-off value for risk was calculated with receiver operating characteristic (ROC) curve analysis. Two validation sets from the prospective cohort were used to validate the risk prediction model by measures including prediction accuracy and the true positive rate. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 327 (54.8%) males with NOA experienced sperm retrieval failure with FNA. FSH level, age and testicular volume were included in the prediction model for sperm retrieval failure risk. The model had an AUC of 82.3% (95% CI: 77.6-87.1%) and a cut-off value of 64.61% with a sensitivity of 0.677 and specificity of 0.863 for predicted risk. The predictive accuracies were 85.25 and 83.56% in the external validation sets from two centres. Specifically, 85.71 and 85.15% of NOA patients from two centres that experienced sperm retrieval failure were correctly identified using our model. LIMITATIONS, REASONS FOR CAUTION: A small proportion of males with NOA in whom sperm were successfully retrieved with FNA were misclassified; therefore, TESA techniques with higher sperm retrieval rates may be attempted in patients with high predicted risks of sperm retrieval failure rather than terminating the efforts to produce a genetic offspring. In addition, the ability to achieve a live birth using sperm retrieved with FNA was not tested in this study. WIDER IMPLICATIONS OF THE FINDINGS: We would recommend the use of micro-TESE for men with NOA and a high predicted risk of FNA failure. STUDY FUNDING/COMPETING INTEREST(S): This study was partly supported by National Key R&D Program of China (No. 2017YFC0907305), the National Natural Science Foundation of China (No.81803332), Sichuan Science & Technology Program (No. 2018SZ0144, 2016SZ0066, 2018SZ0284 and 2018FZ0043), Chengdu Science & Technology Bureau (No. 2018-YF05-01265-SN), Postdoctoral Research foundation of Sichuan University (No. 2018SCU12012) and West China Second University Hospital of Sichuan University (No. kx027). There are no competing interests related to this study. TRIAL REGISTRATION NUMBER: Not applicable.

[Advantages of cryopreservation technology for rare human sperm].

OBJECTIVE: To search for an effective method for cryopreservation of rare human sperm (RHS) by comparing the effect of RHS cryopreservation technology with that of conventional cryopreservation technology on post-thaw sperm from patients with severe oligozoospermia. METHODS: Semen samples obtained from 82 patients with severe oligozoospermia were preserved by RHS cryopreservation technology, and another 24 samples cryopreserved by conventional technology, the former divided into groups A (sperm concentration < 1×106/ml, n = 54) and B (1×106/ml ≤ sperm concentration < 5×106/ml, n = 28), and the latter included in group C (sperm concentration < 15×106 /ml, n = 24). The survival rate of post-thaw sperm and recovery rate of progressively motile sperm (PMS) were compared among the three groups. RESULTS: The survival rate of post-thaw sperm was significantly higher in groups A and B than in C (ï¼»62.8 ± 18.7ï¼½% and ï¼»61.9 ± 17.2ï¼½% vs ï¼»50.7 ± 13.5ï¼½%, P < 0.05), and so was the recovery rate of PMS (ï¼»68.7 ± 18.4ï¼½% and ï¼»70.7 ± 15.5ï¼½% vs ï¼»29.2 ± 12.4ï¼½% , P < 0.05), but there were no statistically significant differences between groups A and B in either of the two parameters (P > 0.05). CONCLUSIONS: The cryopreservation technology for rare human sperm may yield relatively stable post-thaw results and deserves a wide clinical application in preserving male fertility.

Effects of fluorescent light-guided transurethral resection on non-muscle-invasive bladder cancer: a systematic review and meta-analysis.

UNLABELLED: What's known on the subject? and What does the study add? It was well known that FL-guided TURBT could reduce the residual rate of NMIBC, but our systematic review suggested that it was not superior to conventional WL-guided TURBT in diagnostic accuracy and it had no significant effect on short-term RFS and PFS. OBJECTIVE: To assess the diagnostic accuracy and therapeutic outcomes of fluorescent light (FL)-guided transurethral resection (TUR) in non-muscle-invasive bladder cancer (NMIBC). METHODS: A systematic search of PUBMED, EMBASE and Cochrane Library was performed to identify randomized controlled trials comparing the outcomes of FL- and white-light (WL)-guided TUR of bladder tumours (TURBT). Outcomes included tumour detection rate, false-positive diagnosis rate, carcinoma in situ (CIS) detection rate, residual tumour rate, recurrence-free survival (RFS) and progression-free survival (PFS). RevMan 5.1 software was used for the meta-analysis. RESULTS: Data from 14 studies, involving 4078 patients with suspected or proven NMIBC, were pooled and included in the meta-analysis. There was no significant difference in tumour detection rate (relative risk [RR] 0.99; 95% confidence interval [CI] 0.96-1.03; P = 0.64) and CIS detection rate (RR 0.82; 95% CI 0.67-1.02; P = 0.07) between the FL and the WL groups. The false-positive diagnosis rate of the FL group was higher than that of the WL group (RR 0.69; 95% CI 0.49-0.97; P = 0.03). The tumour residual rate was higher in the WL group than in the FL group (RR 2.77; 95% CI 1.47-5.02; P = 0.002). No significant differences were found between groups at 3-month follow-up (RR 1.15; 95% CI 0.79-1.66; P = 0.46) or 12-month follow-up (RR 0.86; 95% CI 0.70-1.06; P = 0.16) or in terms of either RFS or PFS at 12-month (RR 0.99; 95% CI 0.94-1.04; P = 0.57) and 24-month follow-up (RR 1.02; 95% CI 0.98-1.06; P = 0.35). CONCLUSION: FL-guided TURBT was not superior to conventional WL in diagnostic accuracy. Although FL-guided TURBT had an advantage in reducing the residual tumour rate, it had no significant effect on short-term RFS and PFS.

The results of transperineal versus transrectal prostate biopsy: a systematic review and meta-analysis.

This systematic review was performed to compare the efficacy and complications of transperineal (TP) vs. transrectal (TR) prostate biopsy. A systematic research of PUBMED, EMBASE and the Cochrane Library was performed to identify all clinical controlled trials on prostate cancer (PCa) detection rate and complications achieved by TP and TR biopsies. Prostate biopsies included sextant, extensive and saturation biopsy procedures. All patients were assigned to a TR group and a TP group. Subgroup analysis was performed according to prostate-specific antigen (PSA) levels and digital rectal examination (DRE) findings. The Cochrane Collaboration's RevMan 5.1 software was used for the meta-analysis. A total of seven trials, including three randomized controlled trials (RCTs) and four case-control studies (CCS), met our inclusion criteria. There was no significant difference in the cancer detection rate between the sextant TR and TP groups (risk difference (RD), -0.02; 95% confidence interval (CI), -0.08-0.03; P=0.34). Meta-analysis for RCTs combined with CCS showed that there was no difference in the cancer detection rate between the extensive TR and TP group (RD, -0.01; 95% CI, -0.05-0.04; P=0.81). There was no significant difference in PCa detection rate between the saturation TR and TP approaches (31.4% vs. 25.7%, respectively; P=0.3). There were also no significant differences in cancer detection between the TR and TP groups in each subgroup. Although the data on complications were not pooled for the meta-analysis, no significant difference was found when comparing TR and TP studies. TR and TP biopsies were equivalent in terms of efficiency and related complications. TP prostate biopsy should be an available and alternative procedure for use by urologists.