Altered brain activity associated with premature ejaculation improved by electroacupuncture in rats.

Background: Premature ejaculation (PE) is linked with abnormal brain activity that is modifiable by electroacupuncture (EA). Aim: In this study we aimed to explore the central pathological mechanism underlying EA in treating PE. Methods: Six-week-old male Sprague-Dawley rats were divided into a PE group (n = 8) and a control group (n = 8) according to ejaculatory frequency during copulatory behavior. All rats underwent EA at the Zusanli acupoint (ST-36) for 4 weeks. Magnetic resonance imaging data were collected before and after EA. Outcomes: The behavioral parameters, plasma norepinephrine levels, fractional amplitude of low frequency fluctuation (fALFF), and regional homogeneity (ReHo) were evaluated. Results: The PE group ejaculated more times with shorter latency compared with controls. After EA, the ejaculation frequency of the PE group decreased, and the ejaculation latency period increased, with no changes observed in the control group. Norepinephrine levels were higher in the PE group than in the controls and were positively correlated with ejaculation frequency and negatively correlated with ejaculation latency. The PE group showed lower fALFF in the right striatum and higher ReHo in the brainstem compared with controls. After EA, controls showed decreased fALFF in the right striatum, left olfactory bulb, and dorsal fornix and increased ReHo in the right interpeduncular nucleus, as well as decreased ReHo in the left striatum, prelimbic system, right basal forebrain region, septal region, and olfactory bulb, while the model group exhibited increased fALFF in the right hypothalamic region, decreased fALFF in the left globus pallidum and right basal forebrain region and increased ReHo in the right interpeduncular nucleus, as well as decreased ReHo in the left striatum, olfactory bulb, basal forebrain region, dentate gyrus, right dysgranular insular cortex, and striatum. Compared with the controls after EA, the model group showed increased ReHo of the right hypothalamic region and decreased ReHo of the right dysgranular insular cortex. Clinical Implications: These findings might enhance the understanding of PE and contribute to new, targeted therapies for PE. Strengths and Limitations: The therapeutic effects might be achieved by EA inhibiting the activity in brain regions involved in ejaculatory behavior. However, the curative effect of acupuncture might be underestimated due to some curative effects of sham acupuncture used in the control group. Conclusion: In conclusion, the ejaculatory frequency of rats may be reduced and ejaculation latency could be extended by EA at ST-36, which might be achieved by the effects of this treatment on brain activity.

[Analysis of blood flow of penile cavernous artery in 2 568 cases of erectile dysfunction].

OBJECTIVE: To analyze the blood flow parameters of the cavernous arteries of ED patients after injection of vasoactive drugs, and to explore the differences in blood flow of the cavernous arteries in different erectile states. METHODS: Retrospectively analyzed the penile cavernous arterial blood flow parameters of 2568 adult male ED patients after injection of the vasoactive drug (alprostadil). The patients were divided into three groups: maintaining erection group with EHS (erection hardness score) ≥ 3 and sustained erection time ≥ 20 minutes (967 cases), nonpersistent erection group with EHS≥3 and sustained erection time<5 minutes (788 cases), and incomplete erection group with EHS<3 (813 cases). Compared the parameters of age, EHS, duration of erection, cavernous artery peak systolic velocity (PSV), end diastolic velocity (EDV) and resistance index (RI) among the three groups respectively. The maintaining erection group was divided into the youth group (757 cases) which aged less than 40 years old and the middle-aged and elderly group (210 cases) with 40 years old or over. The parameters of PSV, EDV and RI between the two groups were compared. The incomplete erection group were divided into the good blood supply group (407 cases) with the bilateral PSV ≥35cm/s and the insufficient blood supply group (252 cases) with the bilateral PSV<35cm/s. The parameters of age, EHS, EDV and RI between the two groups were compared. RESULTS: The age, PSV, EDV and RI of the three groups were significantly different (P<0.01). In the maintaining erection group, the PSV of the young group was significantly higher than that of the middle-aged and elderly group (P<0.05), but there was no statistically significant difference in EDV and RI (P>0.05). In the incomplete erection group, the EHS, PSV, EDV, and RI of the good blood supply group were significantly higher than those of the insufficient blood supply group (P<0.05), while the age was significantly lower than that of the latter (P<0.01). CONCLUSION: The injection of vasoactive drugs combined with color Doppler ultrasound can directly reflect the blood supply of the cavernous arteries of the penis. The better the erection state, the better the blood supply of cavernous arteries. The middle-aged and elderly people are more likely to have cavernous arteries problem of insufficient blood supply than the young people.

[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.

The Diagnostic Role of Neurophysiological Tests for Premature Ejaculation: A Prospective Multicenter Study.

PURPOSE: Premature ejaculation (PE) is one of the most common male sexual dysfunctions. Local anesthetics (LAs) and dapoxetine are frequently used to treat PE; however, previous studies show variable efficacy. This study aims to determine the efficacy of LAs and dapoxetine using a novel classification based on neurophysiological tests. MATERIALS AND METHODS: This multicenter cohort study enrolled adult men (568) with an intravaginal ejaculatory latency time (IELT) ≤2 minutes. Patients were divided into 4 groups according to the results of neurophysiological tests and assigned different treatments for 12 weeks: 1) penile sensory hyperexcitability type (Sens)-LAs; 2) penile sympathetic hyperexcitability type (Symp)-dapoxetine; 3) mixed type (Mixed)-both LAs and dapoxetine; 4) normal type (Norm)-both LAs and dapoxetine. Self-estimated IELT and patient-reported outcomes were recorded. RESULTS: The total percentage of men achieving IELT >2 minutes and ≥5 minutes after treatment were 82.7% and 76.7%, respectively. For men with abnormal results of neurophysiological tests, 401 (86.6%) had improved IELT >2 minutes after the 12-week treatment course, in which 375 (81.0%) achieved IELT ≥5 minutes. All patient-reported outcome measures improved in each group after 12 weeks of treatment, with greater improvements among those with abnormal neurophysiological tests. CONCLUSIONS: The efficacy of LAs and dapoxetine increased in PE patients with abnormal results of neurophysiological tests. This novel classification of PE using neurophysiological tests could help guide and improve efficacy of PE therapies.

[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.

[PDE-5i combined with RigiScan-based audiovisual sexual stimulation test versus nocturnal penile tumescence test in evaluation of erectile function].

OBJECTIVE: To explore the clinical value of phosphodiesterase type-5 inhibitors (PDE-5i) combined with RigiScan-based audiovisual sexual stimulation (AVSS) test in comparison with that of nocturnal penile tumescence (NPT) test in evaluation of erectile function. METHODS: A total of 166 ED patients, aged 21－63 (mean 31) years, with a disease course of 3 months to 10 years (mean 14 months), underwent NPT test or PDE-5i + RigiScan-based AVSS test from 2017 to 2018. We compared the results of the diagnostic strategies. Normal NPT patterns were presumed to indicate psychogenic and abnormal ones to indicate organic ED. RESULTS: Compared with the results of NPT test, no statistically significant difference was observed in the accuracy rate between Viagra + AVSS test and Cialis + AVSS test (P > 0.05). PDE-5i + RigiScan-based AVSS test achieved a sensitivity of 78.9% and a specificity of 90.7% in the diagnosis of psychogenic ED and an overall accuracy rate of 81.9%. According to the results of PDE-5i + RigiScan-based AVSS test, the patients fell into a normal and an abnormal erection group, with significant differences between the two groups in age, disease course, IIEF-5 score and maintenance time of penile tip rigidity ≥60% (P < 0.05). ROC curve analysis indicated that PDE-5i + RigiScan-based AVSS test accurately manifested the erectile function of the patients. CONCLUSIONS: Compared with NPT test, PDE -5i combined with RigiScan-based AVSS test is simple, inexpensive, practical and with a high sensitivity and specificity, and therefore can be used as the first-choice strategy for etiological diagnosis of ED.

[Penile hemodynamics in different erection phases].

OBJECTIVE: To compare the hemodynamic indexes of penile cavernosal arteries in different phases of penile erection and determine the optimal time for color Doppler ultrasonography of the penis. METHODS: Forty healthy adult male volunteers with normal erectile function received intracavernous injection of 10 µg Alprostadil together with visual and auditory stimuli. Within the next 30 minutes, we monitored dynamically the blood flow in the bilateral penile cavernosal arteries, and recorded and compared the peak systolic velocity (PSV), end diastolic velocity (EDV) and resistance index (RI) in the phases of latency, tumescence, full erection and rigid erection. RESULTS: Fourteen of the subjects experienced latency, tumescence and full erection only, but failed to achieve rigid erection. PSV and RI were significantly higher while EDV remarkably lower in the tumescence and full erection phases than in the latency phase (all P < 0.01). Statistically significant differences were observed between the tumescence and full erection phases in EDV and RI (both P < 0.01) but not in PSV (P > 0.05). Among the 26 males that achieved rigid erection, PSV, EDV and RI showed significant differences between the full and rigid erection phases (all P < 0.01), but not between the left and right cavernosal arteries in the same phase (P > 0.05). CONCLUSIONS: The blood flow in the penile cavernosal arteries changes dynamically in the process of penile erection, and the hemodynamic indexes obtained in the full erection phase can better reflect the function of penile cavernous vessels.

[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.

[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.

Proteomic analysis of N-glycosylation of human seminal plasma.

Seminal plasma is a mixture of secretions from several male accessory glands. The seminal plasma contains many secreted proteins which are important for sperm function and male fertility. In this study, we employed N-linked glycosylated peptide enrichment, combined with LC-MS/MS analysis, and establish the first large scale N-linked glycoproteome of human seminal plasma. Combined with the results of five biological replicates, a total of 720 N-glycosylated sites on 372 proteins were identified. Analysis of variations among five individuals revealed similar compositions of N-glycosylated proteins in seminal plasma. The N-linked glycoproteome could help us understanding the biological functions of human seminal plasma. The data set could also be a resource for further screening of biomarkers for male diseases including cancer and infertility at the level of N-glycosylation. For example, N-glycosylated prostate-specific antigen is known to be an efficient biomarker that can distinguish benign prostate hyperplasia from prostate cancer. All MS data have been deposited in the ProteomeXchange with identifier PXD000959 (http://proteomecentral.proteomexchange.org/dataset/PXD000959).

Insights into the lysine acetylproteome of human sperm.

Protein lysine acetylation is a dynamic and reversible post-modification that is known to play diverse functions in eukaryotes. Nevertheless, the composition and function of non-histone lysine acetylation in gametes remain unknown. In humans, only capacitated sperm have the capacity to fertilize an egg. In the present study, we found complex composition of lysine acetylated proteins in capacitated human sperm. In vitro fertilization inhibition assay by anti-acetyllysine antibody showed essential roles of lysine acetylation in fertilization. And inhibition of lysine deacetylases, the histone deacetylases, by trichostatin A and nicotinamide, could significantly suppress sperm motility. After immunopurification enrichment of acetylpeptides with anti-acetyllysine antibody and high-throughput liquid chromatography-tandem mass spectrometry identification, we characterized 1206 lysine acetylated sites, corresponding to 576 lysine acetylated proteins in human capacitated sperm. Bioinformatics analysis showed that these proteins are associated with sperm functions, including motility, capacitation, acrosome reaction and sperm-egg interaction. Thus, lysine acetylation is expected to be an important regulatory mechanism for sperm functions. And our characterization of lysine acetylproteome could be a rich resource for the study of male fertility. BIOLOGICAL SIGNIFICANCE: Mature sperm are almost transcriptionally and translationally silent, thus post-translational modifications play important roles in sperm functions. Till now, only two types of PTMs, phosphorylation and glycosylation, are well studied in normal human sperm based on large scale proteomics. In the present study, we established the acetylproteome of capacitated human sperm. Over 1000 lysine acetylated sites were identified. Bioinformatics analysis shows that lysine acetylated proteins participate in many biological events of sperm functions. We further provided functional data that the lysine acetylation is essential for sperm motility and fertilization using histone acetylase inhibitors and anti-acetyllysine antibody. These data can be strong evidences for the important function of lysine acetylation in human sperm.