[Semen quality of infertile men carrying hepatitis B virus].

OBJECTIVE: To study the semen parameters of infertile men carrying hepatitis B virus (HBV) and the influence of HBV infection on semen quality. METHODS: We collected the semen samples from 782 infertility males aged 25－35 years old. According to the results of serological examinations, we divided the patients into groups A (HBsAg, HBeAb and HBcAb positive, n = 286), B (HBsAg, HBeAg and HBcAb positive, n = 230) and C (non-HBV control, n = 266), and comparatively analyzed the routine semen parameters, sperm acrosin activity, sperm DNA fragmentation index (DFI) and high DNA stainability (HDS) among the three groups of patients. RESULTS: Compared with the patients of groups B and C, those of group A showed markedly decreased sperm concentration (ï¼»88.20 ± 82.62ï¼½ and ï¼»89.29 ± 53.80ï¼½ vs ï¼»71.49 ± 60.03ï¼½ ×106/ml, P<0.05), progressively motile sperm (PMS) (ï¼»34.88 ± 15.60ï¼½% and ï¼»37.82 ± 13.63ï¼½% vs ï¼»30.70 ± 14.79ï¼½%, P<0.05), sperm motility (ï¼»45.77 ± 16.58ï¼½% and ï¼»48.16 ± 14.03ï¼½% vs ï¼»42.67 ± 17.23ï¼½%, P<0.05), sperm viability (ï¼»82.55 ± 7.55ï¼½% and ï¼»85.26 ± 6.39ï¼½% vs ï¼»81.07 ± 10.19ï¼½%, P>0.05) and morphologically normal sperm (MNS) (ï¼»6.93 ± 4.45ï¼½% and ï¼»7.27 ± 4.43ï¼½% vs ï¼»5.72 ± 3.47ï¼½%, P<0.05), with sperm concentration, PMS, sperm motility, sperm viability and MNS remarkably lower in group B than in C. Sperm acrosin activity was significantly reduced in group A in comparison with groups B and C (ï¼»57.07 ± 26.38ï¼½ vs ï¼»63.03 ± 28.75ï¼½ and ï¼»78.00 ± 33.49ï¼½ µIU/106, P<0.01), remarkably lower in group B than in C (P<0.01). The sperm DFI and HDS, however, were markedly higher in group A than in B (ï¼»14.79 ± 9.46ï¼½% vs ï¼»12.95 ± 7.29ï¼½% and ï¼»11.60 ± 5.98ï¼½%, P<0.05; ï¼»9.62 ± 6.20ï¼½% vs ï¼»8.43 ± 4.72ï¼½% and ï¼»8.41 ± 4.59ï¼½%, P<0.05), and both higher in group B than in C. CONCLUSIONS: Semen quality is lower in infertile men carrying HBV and therefore HBV infection is one of the causes of male infertility.

Integration of hepatitis B virus into patients' sperm genome and its clinical risks.

BACKGROUND: Like the coronavirus disease 2019, the hepatitis B virus is also wreaking havoc worldwide, which has infected over 2 billion people globally. Using an experimental animal model, our previous research observed that the hepatitis B virus genes integrated into human spermatozoa can replicate and express after being transmitted to embryos. However, as of now, this phenomenon has not been confirmed in clinical data from patients. OBJECTIVES: To explore the integration of the hepatitis B virus into patients' sperm genome and its potential clinical risks. MATERIALS AND METHODS: Forty-eight patients with chronic hepatitis B virus infection were categorized into two groups: Test Group-1 comprised 23 patients without integration of hepatitis B virus DNA within the sperm genome. Test Group-2 comprised 25 patients with integration of hepatitis B virus DNA within the sperm genome. Forty-eight healthy male donors were included as control. The standard semen parameter analysis, real-time polymerase chain reaction, quantitative real-time polymerase chain reaction, sperm chromatin structure assay, fluorescence in situ hybridization, and immunofluorescence assays were utilized. RESULTS: The difference in the median copy number of hepatitis B virus DNA per mL of sera between Test Group-1 and Group-2 was not statistically significant. In Test Group-2, the integration rate of hepatitis B virus DNA was 0.109%, which showed a significant correlation with the median copy number of hepatitis B virus DNA in motile spermatozoa (1.18 × 103 /mL). Abnormal semen parameters were found in almost all these 25 patients. The integrated hepatitis B virus S, C, X, and P genes were detected to be introduced into sperm-derived embryos through fertilization and retained their function in replication, transcription, and translation. CONCLUSION: Our findings suggest that hepatitis B virus infection can lead to sperm quality deterioration and reduced fertilization capacity. Furthermore, viral integration causes instability in the sperm genome, increasing the potential risk of termination, miscarriage, and stillbirth. This study identified an unconventional mode of hepatitis B virus transmission through genes rather than virions. The presence of viral sequences in the embryonic genome poses a risk of liver inflammation and cancer.

Changes in human umbilical vein endothelial cells induced by endothelial nitric oxide synthase traffic inducer.

This study investigated the changes in human umbilical vein endothelial cells (HUVECs) induced by overexpression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) and its role in cellular injury. Recombinant NOSTRIN-expressing and empty vectors were transfected into cultured HUVECs, and factor VIII-related antigen was examined by using immunohistochemical analysis. Growth curves were generated for both transfected and untransfected cells and these indicated that the proliferative ability of cells overexpressing NOSTRIN was significantly decreased. The expression of NOSTRIN and eNOS proteins was detected by using Western blot analysis, endothelial NOS (eNOS) activity was assayed by using spectrophotometry, and NO2 (-)/NO3 (-) levels were measured using nitrate reductase. Immunohistochemical analysis demonstrated that all groups expressed NOSTRIN in the plasma membrane and cytoplasm, and Western blot analysis confirmed that NOSTRIN levels were significantly higher in cells transfected with the NOSTRIN plasmid (P<0.01). The activity of eNOS and the levels of NO2 (-)/NO3 (-) were significantly decreased in NOSTRIN overexpressing cells as compared with empty vector and untransfected cells (P<0.01 and P<0.01, respectively). Morphological and ultrastructural changes were observed under light and electron microscopy, and it was found that NOSTRIN-overexpressing cells were elongated with deformities of the karyotheca, injury to the plasma membrane, increased lipids in the cytoplasm, and shortened microvilli. This study showed that overexpression of NOSTRIN had a significant effect on eNOS activity in HUVECs and resulted in significant cellular damage.

[Expression of NOSTRIN in the testis tissue of azoospermia patients].

OBJECTIVE: To determine the expressions of endothelial nitric oxide synthase traffic inducer (NOSTRIN) and endothelial nitric oxide synthase (eNOS) in the testis tissue of azoospermia patients, and investigate their correlation with the pathogenesis of azoospermia. METHODS: We detected the expressions of NOSTRIN and NOSTRIN mRNA by immunohistochemistry and RT-PCR respectively, determined the activity of eNOS by spectrophotometry, and measured the stable metabolic end product NO, NO2- / NO3-, by nitrate reductase assay in the testis tissues of 17 patients with idiopathic azoospermia (the azoospermia group) and 10 normal men (the normal group). RESULTS: NOSTRIN and NOSTRIN mRNA were expressed in the spermatogonia, sertoli cells, stromal cells and vascular endothelial cells, more lowly in the azoospermia than in the normal group (0.312 +/- 0.076 versus 0.793 +/- 0.082, P < 0.01). The activity of eNOS was significantly increased in the idiopathic azoospermia patients ([33.727 +/- 3.58] U/mg) compared with the normal men ([17.69 +/- 3.84] U/mg) (P < 0.01). The level of NO2- / NO3- was significantly higher in the azoospermia than in the normal group ([48.56 +/- 8.49] micromol/L versus [25.37 +/- 9.61] micromol/L, P < 0.01). The expression of NOSTRIN showed a significant negative correlation with the activity of eNOS (r = -0.57, P < 0.01) as well as with the level of NO2- / NO3- (r = -0.61, P < 0.01) in the testis tissue of the idiopathic azoospermia patients. CONCLUSION: The expression of NOSTRIN is decreased, while the activity of eNOS and the level of NO2- / NO3- increased in the testis tissue of azoospermia patients, which may be associated with the pathogenesis of azoospermia.

[Expression of CR16 in the testis of patients with idiopathic azoospermia].

OBJECTIVE: To investigate the expression of corticosteroids and regional expression 16 (CR16) in the testis of patients with idiopathic azoospermia and the role of CR16 in spermatogenesis. METHODS: Immunohistochemistry and RT-PCR were used to detect the expression levels of the CR16 protein and mRNA in the testes of 48 patients with idiopathic azoospermia and 10 healthy men. RESULTS: Immunohistochemistry showed that the CR16 protein expressed in the Sertoli cells and spermatids-binding region in the epithelium of seminiferous tubules. The level of the CR16 protein was markedly lower in the idiopathic azoospermia patients than in the healthy men, and RT-PCR also showed a significantly decreased level of CR16 mRNA in the testis of the patients. CONCLUSION: The expressions of the CR16 protein and mRNA decrease markedly in the testis of patients with idiopathic azoospermia, indicating a correlation with the pathogenesis of azoospermia.