In order to explore the effects of testicular infection of murine cytomegalovirus (MCMV) on mature sperm viability at different periods following MCMV inoculation in mice, 91 BALB/c mice without MCMV infection were randomly divided into two groups: an experimental group (n = 56) and a control group (n = 35). The mice in the experimental group were treated by inoculating MCMV intratesticularly, while those in the controlled group were directly inoculated with DMEM without MCMV. The mice in both groups were sacrificed separately on the day 1, 1. 5, 2, 4, 6, 9 and 14 post-inoculation (D1) 1. 5, 2, 4, 6, 9 and 14 PI). The MCMV M83 mRNA gene was detected in the testis by in situ hybridization (ISH) with MCMV late-mRNA probe labeled with digoxin. Sperm viability of mature sperm in the epididymis cauda was measured. The results demonstrated the positive signal of ISH of MCMV was found mainly in the cytoplasm of the testicular interstitial cells and spermatogenic cells in the experimental group. Compared with that in the controlled group, the sperm viability in the experimental group was decreased significantly on D1 PI and D1.5 PI (P < 0.05). No statistically significant difference in the sperm viability was found after D2 PI between two groups (P > 0.05). This suggested that sperm viability in mice might be descended significantly shortly after MCMV infection and might return to normal with time, indicating that MCMV acute infection might temporarily degrade sperm quality and influence procreation transiently.
Cytomegalovirus Infections/*physiopathology ; Mice, Inbred BALB C ; Orchitis/*virology ; Random Allocation ; Sperm Motility/*physiology ; Spermatozoa/cytology ; Spermatozoa/*physiology

AIMTo study the integration of hepatitis B virus (HBV) DNA into sperm chromosomes in hepatitis B patients and the features of its integration.

METHODSSperm chromosomes of 14 subjects (5 healthy controls and 9 HB patients, including 1 acute hepatitis B, 2 chronic active hepatitis B, 4 chronic persistent hepatitis B, 2 HBsAg chronic carriers with no clinical symptoms) were prepared using interspecific in vitro fertilization between zona-free hamster oocytes and human spermatozoa. Fluorescence in situ hybridization (FISH) to sperm chromosome spreads was carried out with biotin-labeled full length HBV DNA probe to detect the specific HBV DNA sequences in the sperm chromosomes.

RESULTSSpecific fluorescent signal spots for HBV DNA were seen in sperm chromosomes of one patient with chronic persistent hepatitis B. In 9 (9/42) sperm chromosome complements containing fluorescent signal spots, one presented 5 obvious FISH spots and the others 2 to 4 signals. The fluorescence intensity showed significant difference among the signal spots. The distribution of signal sites among chromosomes seems to be random.

CONCLUSIONHBV could integrate into human sperm chromosomes. Results suggest that the possibility of vertical transmission of HBV via the germ line to the next generation is present.

Adult ; Chromosomes, Human ; genetics ; virology ; Hepatitis B ; genetics ; transmission ; virology ; Hepatitis B virus ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Infectious Disease Transmission, Vertical ; Male ; Spermatozoa ; virology ; Virus Integration

OBJECTIVETo study the infection of human cytomegalovirus (HCMV) and herpes simplex virus type II (HSV-I) and the morphological characteristics of the infected spermatogenic cells in the semen of infertile men.

METHODSWe washed and concentrated the spermatogenic cells obtained from 83 semen samples of infertile men, extracted DNA and then screened HCMV and HSV-II by polymerase chain reaction (PCR). Immunocytochemistry (ICC) was used to detect the expression of correlative virus antigens of the positive semen cells, and the cytology smear was employed to observe the morphological changes of the spermatogenic cells under the microscope after cytology staining.

RESULTSOf all the semen samples, 8 were HCMV positive, 4 HSV-II positive, but none were both HCMV and HSV-II positive. HCMV late antigens were positively and HCMV early antigens negatively expressed in the spermatogenic cells of the 8 HCMV positive cases. In the 4 HSV-II positive cases, 3 were positively and 1 weakly positively expressed. In the semen of the 12 positive cases were found large numbers of immature spermatogenic cells, with different manifestations of apoptosis, such as chromatin pycnosis, vacuoles, damaged nuclear membrane, and apoptotic bodies, but without virus infection-induced specific morphological alteration. Sperm concentration of the positive group was significantly lower than that of the negative (P < 0. 05).

CONCLUSIONSpermatogenic cells infected by HCMV and HSV-II may cause pathologic lesions and affect spermatogenesis. Morphologically, the infected spermatogenic cells may undergo some pathologic alteration, such as apoptosis. The rate of HCMV infection is higher among infertile males with pathologic cells in the semen.

Adult ; Antigens, Viral ; analysis ; Cytomegalovirus ; genetics ; immunology ; Cytomegalovirus Infections ; pathology ; virology ; DNA, Viral ; genetics ; Herpes Simplex ; pathology ; virology ; Herpesvirus 2, Human ; genetics ; immunology ; Humans ; Immunohistochemistry ; Infertility, Male ; pathology ; virology ; Male ; Polymerase Chain Reaction ; Semen ; cytology ; virology ; Spermatozoa ; cytology ; virology

AIMTo detect the expression of hepatitis B virus (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization (IVF) technique.

METHODSHuman sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction (PCR) was used to detect HB S and pre-Core/Core (pre-C/C) coding genes both in one- and two-cell embryos. Reverse transcription-PCR (RT-PCR) analysis was used to study the expression of the two genes. Fluorescence in situ hybridization (FISH) analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome.

RESULTSBoth HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences.

CONCLUSIONSperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.

Animals ; Blastula ; virology ; Cricetinae ; DNA Primers ; Female ; Fertilization in Vitro ; Gene Expression Regulation, Viral ; Genome, Viral ; Hepatitis B virus ; genetics ; Humans ; Male ; Mesocricetus ; Oocytes ; physiology ; Ovum ; virology ; Polymerase Chain Reaction ; Semen ; virology ; Spermatozoa ; virology ; Transfection ; Virus Replication ; Zona Pellucida ; physiology

OBJECTIVETo explore the effects of testis murine cytomegalovirus (MCMV) infection on mature sperm viability in mice.

METHODSBALB/c mice without MCMV infection, screened by ELISA, were randomly divided into two groups: an experimental group (n = 64) and a control group (n = 40). The former were directly inoculated with MCMV into the testis, while the latter treated by inoculation of DMEM without MCMV. The mice in both of the groups were sacrificed respectively at 1, 2, 4, 6, 9, 14, 21, 38 d postinoculation (D1, 2, 4, 6, 9, 14, 21, 38 PI), the testis was examined histopathologically, and meanwhile the viability of mature sperms in the epididymis cauda was measured.

RESULTSMCMV basophil inclusion bodies were found in the Leydig cells in the experimental group, and spermatogenic cells were vacuolated and arranged disorderly. Compared with the control group, the sperm viability in the experimental group was decreased significantly by 71.42% to 56.04% (P < 0.05) on D1 PI.

CONCLUSIONThe sperm viability in mice might be descended significantly by MCMV infection in the early period, but restored to normal with time. This shows that MCMV infection might influence procreation transiently.

Animals ; Cell Line ; Cell Survival ; physiology ; Cytomegalovirus Infections ; pathology ; physiopathology ; Male ; Mice ; Mice, Inbred BALB C ; Random Allocation ; Spermatozoa ; physiology ; Testicular Diseases ; pathology ; physiopathology ; virology ; Testis ; pathology

With the development of assisted reproductive technology (ART), more and more hepatitis B virus (HBV)-infected couples have their own children successfully; however,vertical transmission of HBV in ART, especially father-to-child transmission, cannot be avoided. The mechanism of attachment and penetration of HBV into human sperm is still not known. Therefore, understanding the state and mechanism of HBV infection of sperm and the impact of HBV on sperm parameters, following up the ART outcome in man with HBV infection are helpful to solve the fertility problem and to control father-to-child vertical HBV infection.
Female ; Follow-Up Studies ; Hepatitis B ; transmission ; Humans ; Infectious Disease Transmission, Vertical ; Male ; Pregnancy ; Pregnancy Outcome ; Reproductive Techniques, Assisted ; Spermatozoa ; virology

OBJECTIVETo investigate the effects of hepatitis B virus (HBV) in semen on human semen parameters and sperm DNA integrity.

METHODSWe detected HBV DNA in the semen samples of 153 HBsAg-seropositive patients by real-time fluorescence quantitative PCR and calculated the sperm nuclear DNA fragmentation index (DFI) by sperm chromatin dispersion (SCD) assay. We compared the semen parameters between the HBV DNA-positive group (A, n = 43) and HBV DNA-negative group (B, n = 110) and analyzed the correlation of sperm DFI with the number of HBV DNA copies in the semen.

RESULTSHBV DNA was detected in 43 (28.1%) of the 153 semen samples. No statistically significant differences were observed in age, semen volume and sperm concentration between groups A and B (P >0.05). Compared with group B, group A showed significantly decreased sperm viability ([58.0 +/- 18.8]% vs [51.4 +/-17.1]%, P<0.05), progressively motile sperm ([29.6 +/- 13.3]% vs [24.5 +/- 10.1]%, P<0.05), average straight-line velocity ([23.7 +/- 4.0] microm/s vs [19.9 +/- 4.5 ] microm/s, P<0.01) and average path velocity ([26.5 +/- 7.0] microm/s vs [23.4 +/- 5.3] microm/s, P<0.01), but remarkably decreased sperm DFI ([19.3 +/- 8.0]% vs [24.2 +/- 9.4]%, P<0.01). The number of HBV DNA copies in semen exhibited a significant positive correlation with sperm DFI (r = 0.819, P < 0.01).

CONCLUSIONHBV DNA in semen is not significantly associated with the number of sperm, but may affect sperm viability, velocity and DFI. There is a load-effect relationship between the number of HBV DNA copies in semen and sperm nuclear DNA integrity.

Adult ; DNA Fragmentation ; DNA, Viral ; isolation & purification ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; Humans ; Male ; Semen Analysis ; Sperm Count ; Spermatozoa ; virology

OBJECTIVETo investigate sperm DNA integrity in male infertility patients with hepatitis B virus (HBV) infection.

METHODSThis study included 90 infertile men with HBV infection (group A), 82 infertile men without HBV infection (group B) and 70 normal fertile men (group C). We detected sperm DNA integrity among the subjects, including DNA fragmentation index (DFI) and high DNA stainability (HDS), by sperm chromatin structure assay (SCSA), and compared them among the three groups.

RESULTSDFI was higher in group A ([28.17 +/- 13.06]%) than in B ([26.64 +/- 9.79]%) and C ([15.67 +/- 4.73]%), significantly higher in A and B than in C (P < 0.05) but with no significant difference between A and B (P > 0.05). HDS was higher in group A ([10.83 +/- 5.601]%) than in B ([9.04 +/- 3.48]%) and C ([8.04-2.25]%), with significant difference between A and C (P < 0.05).

CONCLUSIONSperm DNA integrity of infertile males is significantly different from that of normal fertile men, and infertility with HBV infection further impairs sperm DNA, which is manifested by abnormal sperm nuclear maturity.

Adult ; Case-Control Studies ; Chromatin ; DNA ; genetics ; DNA Damage ; Hepatitis B ; pathology ; Hepatitis B virus ; Humans ; Infertility, Male ; genetics ; virology ; Male ; Sperm Count ; Spermatozoa ; pathology ; Young Adult

ObjectiveTo evaluate the semen quality of the HIV/AIDS male patients after treated by the highly active antiretroviral therapy (HAART) and their potential of transmitting HIV/AIDS and provide some evidence for this cohort of males who wish for parenthood.

METHODSWe collected semen samples from 20 HIV/AIDS male patients who had been treated by HAART for over 6 months and wished for parenthood. We examined sperm concentration, viability and total motility and the percentage of morphologically normal sperm (MNS) using the computer-assisted semen analysis system, measured the HIV-1 RNA loads in the semen by the Cobas Amplicor Monitor test, and counted CD4+ T cells in the peripheral blood by flow cytometry.

RESULTSThe patients were aged 25－40 (30.7 ± 5.05) years. After treated by HAART for 6－26 (14.24 ± 12.26) months, the count of blood CD4+ T cells was significantly increased (341－1 058 ［535.76 ± 212.021］ /μl) in comparison with the baseline (226－965 ［422.38 ± 200.86］ /μl). Compared with the normal value, the semen volume was increased except in 1 case (≥2 ml) while total sperm motility was decreased in 13 cases (≥40%), and so were sperm concentration in 2 cases (≥15 × 106 / ml), sperm viability in 5 (58%), the percentage of progressively motile sperm in 18 (≥32%), and the percentage of MNS in 6 (≤4%). HIV-1 RNA in the peripheral blood was <20 copies/mL in all the cases and that in the seminal plasma was also <20 copies/ml in 18 cases but >20 copies/mL in the other 2 (［4.70 × 101］ and ［2.2 × 102］ copies/ml, respectively). Of the 4 couples that had sex without protective measures for over 6 months, all the 4 female partners exhibited negative HIV antibodies in regular follow-up examinations and 1 achieved spontaneous pregnancy and healthy birth, with negative HIV-1 RNA in both the mother and the baby.

CONCLUSIONSThe HIV RNA level is higher in the semen than in the blood of the HIV/AIDS male patients after HAART, which indicates the potential risk of their semen transmitting HIV/AIDS to their female partners. Their sperm concentration and total sperm motility are lower than the normal value, which suggests a decreased fertility.

Adult ; Antiretroviral Therapy, Highly Active ; Female ; Flow Cytometry ; HIV Infections ; drug therapy ; virology ; Humans ; Male ; Pregnancy ; RNA, Viral ; analysis ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Young Adult

The novel coronavirus disease (COVID-19) pandemic is emerging as a global health threat and shows a higher risk for men than women. Thus far, the studies on andrological consequences of COVID-19 are limited. To ascertain the consequences of COVID-19 on sperm parameters after recovery, we recruited 41 reproductive-aged male patients who had recovered from COVID-19, and analyzed their semen parameters and serum sex hormones at a median time of 56 days after hospital discharge. For longitudinal analysis, a second sampling was obtained from 22 of the 41 patients after a median time interval of 29 days from first sampling. Compared with controls who had not suffered from COVID-19, the total sperm count, sperm concentration, and percentages of motile and progressively motile spermatozoa in the patients were significantly lower at first sampling, while sperm vitality and morphology were not affected. The total sperm count, sperm concentration, and number of motile spermatozoa per ejaculate were significantly increased and the percentage of morphologically abnormal sperm was reduced at the second sampling compared with those at first in the 22 patients examined. Though there were higher prolactin and lower progesterone levels in patients at first sampling than those in controls, no significant alterations were detected for any sex hormones examined over time following COVID-19 recovery in the 22 patients. Although it should be interpreted carefully, these findings indicate an adverse but potentially reversible consequence of COVID-19 on sperm quality.
Adult ; Asthenozoospermia/virology* ; COVID-19/physiopathology* ; China ; Gonadal Steroid Hormones/blood* ; Humans ; Male ; Progesterone/blood* ; Prolactin/blood* ; SARS-CoV-2 ; Semen/physiology* ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa/physiology* ; Time Factors