[Differential expression profiles of miRNAs in the vaginal tissue of the women with lubrication disorders].

OBJECTIVE: To explore the pathogenesis of lubrication disorder (LD), a most common type of female sexual dysfunction affecting women's physical health and conception, and find the therapeutic targets for its treatment and prevention. METHODS: We chose 3 LD patients and 3 healthy controls in Nanjing Maternal and Child Health Hospital, extracted their vaginal epithelial RNA for high-throughput miRNA sequencing, screened differentially expressed miRNAs for hierarchical cluster analysis, target gene prediction and gene ontology (GO) and KEGG pathway enrichment analyses. Finally, we verified the sequencing results by real-time fluorescent quantitative PCR. RESULTS: Totally 1 673 miRNAs were predicted by high-throughput sequencing and 64 likely to be the targets for the treatment of LD were screened, including 25 up-regulated more than 4 times and 39 down-regulated more than 4 times in the LD patients compared with the healthy controls. The neuron projection morphogenesis and AMPK signaling pathway were the most significant enrichment GO term and KEGG pathway. CONCLUSIONS: miRNAs are expressed differentially in LD patients. These miRNAs and target genes may be related to the occurrence of LD, and those that are expected to be the targets for the treatment of LD have important theoretical significance and potential application value.

[HPV infection of the external genitalia in men whose female partners have cervical HPV infection].

OBJECTIVE: Human papilloma virus (HPV) is a necessary cause of cervical cancer and is also closely related to penile cancer, oropharyngeal cancer, and anal cancer in males. However, few studies are reported on male HPV. This study aimed to investigate HPV infection of the external genitalia in men whose female partners have cervical HPV infection. METHODS: We collected the relevant data on the male outpatients whose partners had cervical HPV infection in our Department of Urology and Andrology from August to December 2016. We obtained samples with nylon swabs from the glans penis, corona, inner layer of the prepuce and penile body and detected different types of HPV infection using the Hybribio HPV typing kit, PCR and membrane hybridization. RESULTS: Valid data were collected from 140 males, which showed 83.5% of HPV infection of the external genitalia, including 60 cases of HPV6 (43.2%), 27 cases of HPV16 (19.4%), 14 cases of HPV39 (10.1%), 13 cases of HPV18 (9.4%), 13 cases of HPV58 (9.4%), and 13 cases of HPV52 (9.4%). Redundant prepuce was found in 75.5% of the males, but there was no statistically significant difference in the incidence rate of HPV infection between the normal and redundant prepuce groups (P > 0.05). CONCLUSIONS: Men who have the female partners with positive cervical HPV are at high risk of HPV infection and therefore need to be screened and treated so as to reduce HPV infection in both sexes.

[Relationship of abnormal sperm DNA methylation with early spontaneous abortion].

OBJECTIVE: To investigate the relationship between the abnormal sperm DNA methylation level and early spontaneous abortion. METHODS: We randomly selected 98 males who met the inclusion criteria and whose wives suffered from unexplained abortion or embryo abortion, and included another 46 normal healthy men present for pre-pregnancy check-up as controls. We examined the semen quality and sperm morphology, obtained the sperm DNA fragmentation index (DFI) by modified sperm chromatin dispersion, and measured the sperm DNA methylation level using the methylated DNA quantification kit and the colorimetric method. RESULTS: Compared with the normal controls, the men in the unexplained abortion group showed a significantly lower rate of big-halo sperm (ï¼»45.50 ± 26.27ï¼½ vs ï¼»36.49 ± 23.06ï¼½%, P = 0.038), a higher rate of abnormal-head sperm (ï¼»77.08± 12.21ï¼½ vs ï¼»81.09± 10.89ï¼½%, P = 0.049), and a lower level of sperm DNA methylation (ï¼»0.47 ± 0.33ï¼½ vs ï¼»0.36 ± 0.26ï¼½ ng/µl, P = 0.035). The sperm DNA methylation level was positively correlated with the percentage of big-halo sperm (OR=0.546, P<0.01). Multivariate regression analysis manifested that sperm head abnormality was an independent risk factor of early spontaneous abortion or embryo abortion (OR=1.032, P = 0.049), while the high methylation level was protective factor against early spontaneous abortion or embryo abortion (OR=0.244, P = 0.03). CONCLUSIONS: The abnormal level of sperm DNA methylation may be one of the important reasons for early spontaneous abortion or embryo abortion.

[Relationship between epigenetics of sperm and embryogenesis].

Epigenetics comprises the modifications made in gene expressions without changing the DNA sequence itself. Significant epigenetic changes take place during spermatogenesis and fertilization and exert direct influences on embryogenesis. This article provides an overview of the latest researches on epigenetics of male germ cells and a brief discussion on the correlation of sperm with embryogenesis in four aspects: DNA methylation, histone modification, regulation of non-coding RNAs, and genomic imprinting.

[Protective effect of pAdxsi-ERbeta adenovirus transfection on penile vascular endothelium in ERbetaKO mice].

OBJECTIVE: To investigate the effect of the overexpression of the ERbeta gene on the penile vascular endothelium of ERbeta knockout (ERbetaKO) mice and its molecular mechanisms. METHODS: We randomly divided 12 ERbetaKO male mice into groups A (ERbetaKO + TNFalpha + pAdxsi-ERbeta) and B (ERbetaKO + TNFalpha + empty virus), the former treated by pAdxsi-ERbeta adenovirus transfection, the latter with empty virus, and meanwhile both injected intraperitoneally with TNFalpha at 6 microg per kg body weight per d for 14 days. Then we observed the erectile function of the mice by APO, determined the changes of the endothelial markers CD34 and vWF by immunohistochemical staining, and detected the expressions of the relevant molecules in the eNOS-NO pathway by RT-PCR, Western blot and immunohistochemistry. RESULTS: Compared with group B, group A showed a significantly increased number of penile erections (0.50 +/- 0.55 vs 2.17 +/- 0.41, P < 0.05), shortened erectile latency ([28.83 +/- 1.33] min vs [24.00 +/- 1.27] min, P < 0.05), enriched CD34 and vWF markers (0.67 +/- 0.52 vs 1.50 +/- 0.55 and 0.50 +/- 0.55 vs 1.33 +/- 0.52, both P < 0.05), elevated expressions of eNOS and Cam (RT-PCR: 1.38 +/- 0.03 vs 1.62 +/- 0.05 and 1.02 +/- 0.09 vs 1.42 +/- 0.05, both P < 0.05; Western blot: 1.27 +/- 0.04 vs 1.55 +/- 0.07 and 0.76 +/- 0.05 vs 0.95 +/- 0.08, both P < 0.05), and reduced expression of caveolin-1 (RT-PCR: 2.13 +/- 0.13 vs 1.72 +/- 0.08, P < 0.05; Western blot: 3.99 +/- 0.16 vs 3.40 +/- 0.14, P < 0.05). The results of RT-PCR were consistent with those of Western blot. CONCLUSION: The ERbeta gene protects the penile vascular endothelium via the eNOS-NO pathway.

[Protective effect of ERbeta on penile vascular endothelium in mice].

OBJECTIVE: To investigate the protective effect of ERbeta on the penile vascular endothelium in mice. METHODS: We randomly selected 12 ERbeta knockout (ERbetaKO) and 12 C57BL/6 male mice, and divided them into four groups: normal control, ERbetaKO, ERbetaKO + TNFalpha, and wild-type + TNFalpha group. The former two were treated with normal saline, while the latter two by intraperitoneal injection of TNFalpha at 6 microg/(kg x d) for 14 days. Then we observed the spontaneous erectile response induced by APO and changes of the endothelial cells by immunohistochemical staining of CD34 and vWF, and detected cell apoptosis in the penile cavernous tissue by TUNEL. RESULTS: Compared with the normal control group, the ERbetaKO group showed significantly increased erectile latency (P<0.05), but no significant difference in the number of erections; the ERbetaKO + TNFalpha and wild-type + TNFalpha groups, too, exhibited remarkably longer erectile latency (P<0.05) but fewer erections (P<0.05), with even more obvious changes in the ERbetaKO + TNFalpha group. The expressions of CD34 and vWF were significantly reduced in the ERbetaKO group (2.25 +/- 0.50 and 2.00 +/- 0.00), ERbetaKO + TNFalpha group (0.25 +/- 0.50 and 0.33 +/- 0.58) and wild-type + TNFalpha group (1.50 +/- 0.58 and 1.25 +/- 0.50) as compared with those in the control (3.00 +/- 0.00 and 2.75 +/- 0.50) (P<0.05), even lower in the ERbetaKO + TNFalpha than in the wild-type + TNFalpha group (P<0.05). Apoptotic cells were found only in the ERbetaKO + TNFalpha group. CONCLUSION: After ERbeta knockout and especially after treated with the endothelial injury factor TNFalpha, endothelial cells are decreased in the penile vessels in mice, which suggests that ERbeta has a protective effect on the penile cavernous sinus endothelium.

[Endothelial injury and erectile dysfunction].

The endothelium plays an important role in maintaining vascular homeostasis, regulating vascular tone and blood flow, and preserving a non-thrombogenic blood-tissue interface, and the normal function of the vascular endothelium is essential for penile erection. In most cases, erectile dysfunction (ED) is accompanied by endothelial dysfunction, and endothelial injury is a major pathological basis of ED, which can be induced by bad lifestyles, cardiovascular diseases, reactive oxygen species, and inflammatory mediators. The vascular endothelium is capable of self-repairing, and endothelial injury results from the unbalanced factors of injury and repair. This review focuses on the mechanism and repair of endothelial injury and the relationship of endothelial injury with ED.

[Effects of L-carnitine on the apoptosis of spermatogenic cells and epididymal sperm count and motility in rats with diabetes mellitus].

OBJECTIVE: To explore the effects of L-carnitine (LC) on the apoptosis of spermatogenic cells and on the count and motility of epididymal sperm in rats with diabetes mellitus (DM). METHODS: Twenty-four SD rats (200-230 g) were randomly divided into a control group, a DM model group and an LC group. After the establishment of DM models in the latter two groups by injection of streptozotocin (STZ) at 65 mg/kg, the controls and DM models were treated intragastrically with physiological saline, while the rats in the LC group with LC at 300 mg/kg, all for 6 consecutive weeks. Twenty-four hours after the last administration, all the rats were killed for the detection of the count and motility of epididymal sperm and the apoptosis of spermatogenic cells. RESULTS: The motilities of caput and cauda epididymal sperm were (53.7 +/- 1.8)% and (60.3 +/- 1.6)% in the LC group, significantly higher than in the DM model group ([32.2 +/- 2.0]% and [40.5 +/- 1.4]%, P < 0.05), but remarkably lower than in the control ([63.1 +/- 2.4 ]% and [68.9 +/- 1.3]%, P < 0.05). The count of cauda epididymal sperm was (25.5 +/- 1.1) x 10(6)/100 mg in the DM models, and was increased to (32.0 +/- 1.5) x 10(6)/100 mg after LC treatment (P < 0.05), but still markedly lower than in the controls ([37.8 +/- 1.1] x 10(6)/100 mg) (P < 0.05). The apoptosis rate of spermatogenic cells was (52.5 +/- 4.4)% in the DM model group, and it was reduced to (35.3 +/- 3.5)% after LC administration (P < 0.05), but still significantly higher than in the control group ([3.7 +/- 1.3]%) (P < 0.05). CONCLUSION: Intragastrically gavage of LC at 300 mg/kg for 6 weeks increased the epididymal sperm count, improved sperm motility, and reduced the apoptosis of spermatogenic cells in rats with DM.

[Flow cytometry for evaluation of sperm plasma membrane integrity in varicocele patients].

OBJECTIVE: To detect the sperm plasma membrane integrity (PMI) of varicocele (VC) patients using SYBR-14/PI fluorescent staining and flow cytometry, and investigate its clinical significance. METHODS: We collected semen samples from 120 men, including 30 grade-1 varicocele patients (VC1), 30 grade-2 (VC2), 30 grade-3 (VC3), and 30 normal fertile volunteer controls. Conventional semen analyses were performed by computer-assisted semen analysis (CASA). All the semen samples were washed with PBS and then subjected to SYBR-14/PI staining for the detection of sperm PMI by flow cytometry. The proportion of normal sperm with PMI was indicated as the percentage of sperm emitting green fluorescence (SYBR-14+/PI- %), sperm PMI was determined and sperm fertilization capacity predicted. RESULTS: Significant differences were detected in SYBR-14+/PI- and SYBR-14-/PI+ between the normal men and varicocele male patients (P < 0.01). The percentages of the sperm with PMI (SYBR-14+/PI- %) were remarkably lower in the VC1, VC2 and VC3 groups ([54.85 +/- 3.78]%, [45.37 +/- 4.12]% and [35.14 +/- 4.91]%) than in the normal controls ([70.79 +/- 6.71]%). SYBR-14+/PI-% was correlated positively with sperm motility (r=0.965, P < 0.01) and the percentage of grade a + b sperm (r = 0.874, P < 0.01), negatively with the percentage of grade d sperm (r = -0.965, P <0.01), but not significantly with pH, semen volume and liquefaction time (P > 0.05). CONCLUSION: SYBR-14/PI fluorescent staining and flow cytometry can quickly and exactly detect sperm PMI. Varicocele decreases sperm PMI, which might be an important cause of male infertility.