[Comparative analysis of two assaysin detection of sperm DNA fragmentation index, flow cytometry and AI-based fluorescence microscopy, based on AO staining: A multicentre study].

OBJECTIVE: To study the correlation, consistency, and variations between two assays of DNA fragmentation index based on acridine orange (AO) staining via AI-based fluorescence microscopy(AI-DFI), and flow cytometry (FCM-DFI) across multiple centers. METHODS: We selected 421 male patients from Nanjing Drum Tower hospital ( Hospital G) (226 cases), Eastern Theatre General Hospital (Hospital J) (89 cases) and Jiangsu Province Hospital (Hospital S) (106 cases) . Semen samples from each patient were analyzed for routine semen parameters and for DFI using both AI fluorescence microscopy and flow cytometry. We studied the two methods' stability as well as the correlation, consistency, and variation between the two methods' results in various centers. RESULTS: The two replicate studies' results of AI-DFI and the three centers' FCM-DFI for linear regression analysis indicated strong stability (R2>0.9).Overall(Group A), the AI-DFI results demonstrated good correlation and consistency with the FCM-DFI results of three centers (r>0.85；ICC>0.9).The semen specimens were categorized into two groups: normal specimen group (group B) and abnormal specimen group (group C) (including asthenozoospermia, oligospermia, and semen samples with high impurities).Group C's results showed a decline in correlation and consistency when compared to group A and group B, whereas group B's results showed a little rise in correlation and consistency when compared to group A. Although the consistency and correlation between the results of the two DFI testing methods in the three centers were good, there was still a significant difference between Groups A and C (P<0.05), and in Group B there was a significant difference between the two DFI testing methods only in Hospital G (p＝0.02), with no significant difference in Hospitals J and S (P> 0.05). CONCLUSION: The two detection methods exhibit good stability and correlation. However, significant differences are observed in the DFI detection methods in samples with abnormal semen parameters and high complexity. The main reason for these significant differences may lie in the variations in detection principles. Each detection method has its own advantages, allowing clinical or research settings to choose between them based on laboratory conditions or specific requirements.

[Artificial intelligence-based fluorescence method versus traditional flow cytometry in detection of sperm DNA fragmentation index].

OBJECTIVE: To compare the result of the artificial intelligence (AI) recognition-based fluorescence method and that of traditional flow cytometry in the examination of the sperm DNA fragmentation index (DFI) and assess the reliability of the AI-based fluorescence detection. METHODS: Using flow cytometry and the AI-based fluorescence method, we examined the sperm DFI in the semen samples collected from 338 outpatients. We analyzed the correlation between the results and compared the positive rates detected by the two methods. We repeated the AI-based fluorescence method twice for each semen sample to observe its technical stability in the detection of sperm DFI. RESULTS: The result of flow cytometry was well correlated with that of the AI-based fluorescence method in the detection of sperm DFI (R2 = 0.7131), but poorly correlated for low-concentration, sticky semen and some other extreme samples (R2 = 0.2065). No statistically significant difference was found between the two methods in the positive rate of detection. The AI-based fluorescence method exhibited an excellent technical stability (R2 = 0.9671). CONCLUSION: The AI-based fluorescence method has an excellent technical stability in the detection of sperm DFI and the result is not significantly different from that of traditional flow cytometry.

[Impacts of different Y chromosome microdeletions on spermatogenesis].

OBJECTIVE: To study the semen parameters of the patients with Y chromosome microdeletions and their impacts on the spermatogenesis of the patients. METHODS: We selected 151 male infertility patients with Y chromosome microdeletions from those diagnosed and treated in our hospital and retrospectively analyzed the influence of their semen parameters on the spermatogenic function. RESULTS: Of the 151 cases of Y chromosome microdeletions, AZFc was involved in 102 (66.89%), AZFb in 6 (3.97%), AZFa in 5 (3.31%), AZFa+c in 1 (0.66%), AZFb+c in 6 (3.97%), AZFc+d in 1 (0.66%), AZFb+c+d in 13 (8.61%), AZFa+b+c+d in 12 (7.95%), sY127 in 3 (1.99%), sY134 in 1 (0.66%) and sY86 in 1 (0.66%). Among the total number of the infertility patients, 48 (31.78%) were diagnosed with azoospermia, 74 (49%) with cryptozoospermia, 28 (18.54) with oligoasthenozoospermia and 1 (0.66%) with asthenoteratozoospermia. CONCLUSIONS: Y chromosome microdeletions may lead to decreased sperm quality, and different types of deletion have different impacts on the spermatogenic function of the patients.

[Influence of mycoplasma genitalium infection on semen parameters and seminal plasma biochemical indicators in male infertility patients].

OBJECTIVE: To investigate the influence of mycoplasma genitalium (MG) infection on semen parameters and seminal plasma biochemical indicators in infertile men and the relationship of MG infection with male infertility. METHODS: This retrospective study included 420 male patients with idiopathic infertility confirmed in our hospital from February 2016 to February 2018. We examined the MG RNA in the urine of the patients by nucleic acid amplification test, analyzed the semen parameters using the computer-assisted semen analysis system, observed the sperm morphology by modified Shorr staining, and determined the activities of α-glucosidase (α-Glu), fructose (Fru), zinc and γ-L-glutamyl transpeptidase (γ-GT) in the seminal plasma. RESULTS: Of the 420 cases of idiopathic infertility, 101 were MG-positive and the other 319 MG-negative. Compared with the MG-negative patients, the MG-positive group showed a remarkably decreased semen volume (ï¼»3.57 ± 1.36ï¼½ vs ï¼»3.20 ± 1.30ï¼½ ml, P = 0.016) but no statistically significant differences in sperm concentration (ï¼»57.36 ± 40.88ï¼½ vs ï¼»54.80 ± 36.54ï¼½ ×106/ml, P > 0.05), the percentage of progressively motile sperm (ï¼»45.33 ± 20.42ï¼½% vs ï¼»41.29 ± 18.71ï¼½%, P > 0.05) and the percentage of morphologically normal sperm (ï¼»5.87 ± 2.97ï¼½% vs ï¼»5.67 ± 2.86ï¼½%, P > 0.05). Nor were there any significant differences between the MG-negative and -positive groups in the activities of α-Glu (ï¼»338.82 ± 126.36ï¼½ vs ï¼»352.47 ± 213.34ï¼½ U/L, P > 0.05), Fru (ï¼»15.62 ± 6.35ï¼½ vs ï¼»14.93 ± 6.53ï¼½ mmol/L, P > 0.05), zinc (ï¼»2.82 ± 1.23ï¼½ vs ï¼»2.98 ± 1.30ï¼½ mmol/L, P > 0.05), and γ-GT (ï¼»1993.98 ± 556.03ï¼½ vs ï¼»1925.64 ± 593.41ï¼½ U/L, P > 0.05) in the seminal plasma. CONCLUSIONS: MG infection can reduce the semen volume but has no significant influence on the other semen parameters and seminal plasma biochemical indicators in male infertility patients.

[Cell suspension examination versus histopathological technique in detecting sperm in the testis tissue of non-obstructive azoospermia patients undergoing testicular sperm aspiration].

OBJECTIVE: To explore the clinical selection and application of cell suspension examination (CSE) or histopathological technique (HPT) in detecting sperm in the testis tissue obtained by testicular sperm aspiration (TESA) in patients with non-obstructive azoospermia (NOA). METHODS: Totally, 1 006 NOA patients underwent TESA and their testis tissues were subjected to CSE or HPT for sperm detection. Based on the results of CSE, the testicular tissue samples were divided into groups A (with sperm, n = 567) and B (without sperm, n = 439) and the results were compared with those of HPT. RESULTS: HPT showed 508 cases with but 59 without sperm in group A, and 403 with and 36 without sperm in group B. The consistency rate of CSE with that of HPT was 90.56% (Kappa =0.809), and CSE exhibited a significantly higher rate of sperm detection than HPT (56.36% vs 54.08%, P=0.023). CONCLUSIONS: CSE combined with HPT for detecting sperm in the testis tissue of NOA patients undergoing diagnostic TESA helps clinical diagnosis and treatment. The results of CSE have a decisive significance for assisted reproductive therapy, while those of HPT may provide some definite etiological evidence for drug therapy or surgery.