Objective: To explore the protective effect of Tongjingling (TJL) against sperm DNA damage and oxidative stress in the rat model of experimental varicocele (EVC). METHODS: We randomly divided 75 Wistar male rats into five groups of equal number: sham operation, EVC model, high-dose TJL, mid-dose TJL, and low-dose TJL. The EVC model was established in the rats by partial ligation of the left renal vein, followed by 8 weeks of medication from the 4th week after modeling. Then we observed the general status of the rats, detected the sperm DNA fragmentation index (DFI) in the epididymis by sperm chromatin structure assay (SCSA), and measured the content of hydroperoxide (H2O2) and the activities of catalase (CAT) and superoxide dismutase (SOD) in the testis by colorimetry. RESULTS: Compared with the sham operation group, the EVC models showed significantly increased sperm DFI in the epididymis (P <0.01) and elevated level of H2O2 and activities of CAT and SOD in the testis (P <0.01). In comparison with the EVC models, the rats of the TJL groups exhibited remarkably reduced sperm DFI and H2O2 content, but increased activities of SOD and CAT. CONCLUSIONS TJL can improve sperm DNA integrity by increasing the activities of SOD and CAT and reducing the H2O2 level and hence oxidative stress in the testis tissue.
Animals ; Catalase ; analysis ; DNA ; drug effects ; DNA Fragmentation ; Drugs, Chinese Herbal ; pharmacology ; Epididymis ; chemistry ; Humans ; Hydrogen Peroxide ; analysis ; Ligation ; Male ; Oxidative Stress ; Random Allocation ; Rats ; Rats, Wistar ; Spermatozoa ; Superoxide Dismutase ; analysis ; Testis ; chemistry ; drug effects ; Varicocele ; etiology ; genetics ; metabolism

OBJECTIVETo study the relationship of varicocele (VC) with the expressions of T-type channel alpha1H and alpha1G in the sperm of VC patients.

METHODSBased on the WHO criteria, we examined the semen samples by computer-aided sperm analysis (CASA), and divided the samples into groups A (normal semen from volunteers, n = 20), B (normal semen from VC patients, n = 16) and C (abnormal semen from VC patients, n = 44). We optimized the semen by discontinuous Percoll grade centrifugation, and determined the mRNA expressions of T-type channel alpha1H and alpha1G in the three groups using using reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with group A, the mRNA expressions of alpha1H and alpha1G showed with no significant decrease in group B (P>0.05), but were remarkably reduced in group C (P<0.01).

CONCLUSIONThe abnormal mRNA expressions of T-type channel alpha1H and alpha1G may be one of the causes of declined semen quality and consequently infertility in VC patients, which has pointed out a new direction for the studies of the causes and treatment of VC-related infertility.

Adolescent ; Adult ; Calcium Channels, T-Type ; genetics ; metabolism ; Case-Control Studies ; Humans ; Infertility, Male ; genetics ; Male ; RNA, Messenger ; genetics ; Semen Analysis ; Spermatozoa ; metabolism ; Varicocele ; genetics ; metabolism ; Young Adult

OBJECTIVETo investigate the relationship between Y chromosome microdeletions and human spermatogenesis in infertile men with varicocele (VC).

METHODSWe divided 174 infertile VC patients into groups A (with azoospermia, n = 47) , B (with severe oligozoospermia, n=57) and C (with mild oligozoospermia, n=70), and enlisted 28 fertile males and 26 fertile females as normal controls. We collected DNA from the peripheral blood, amplified 6 sequence tagged sites in AZFa, AZFb and AZFc using multiplex PCR technique. Then we separated and scanned the amplified products by agarose gel electrophoresis to identify microdeletions and their types in comparison with the controls.

RESULTSY chromosome microdeletions were observed in 12.64% of the patients (22/174), 11 cases in group A and the other 11 in group B, but none in group C and the normal controls. The differences were statistically significant (P<0.05). In group A, 6 of the microdeletion cases were in the AZFc region, 1 in the AZFa region, 2 in the AZFb region and 2 in both AZFb and AZFc regions, while in group B, 8 cases were in the AZFc region, 2 in the AZFb region and 1 in both AZFb and AZFc regions.

CONCLUSIONInfertility is correlated to Y chromosome microdeletions in VC patients. Y chromosome microdeletion screening should be performed for infertile VC patients, especially for those with azoospermia or severe oligozoospermia.

Adult ; Case-Control Studies ; Chromosome Deletion ; Chromosomes, Human, Y ; Female ; Humans ; Infertility, Male ; genetics ; Male ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; Varicocele ; genetics

OBJECTIVETo investigate Y chromosome microdeletions in severe oligospermia men with varicocele.

METHODSWe randomly selected 100 cases of severe oligospermia with left varicocele (sperm concentration <5 x 10(6)/ml, group 1), 100 cases of mild oligospermia with left varicocele (sperm concentration 10 -20 x 10(6)/ml, group 2), 100 cases of idiopathic infertility with severe oligospermia (group 3), 100 cases of idiopathic infertility with moderate oligospermia (group 4) and 30 normal fertile men as controls (group 5). We used polymerase chain reaction (PCR) technology to screen 9 sequence tagged sites (STS) of the AZF a, b and c regions and detect Y chromosome microdeletions.

RESULTSAZF microdeletions were found in 19 patients in group 1 (19%) and 11 in group 3 (11%), with a higher rate in the former than in the latter, but not in the other three groups.

CONCLUSIONScreening of Y chromosome microdeletions should be performed before the treatment of severe spermatogenesis with varicocele.

Adult ; Chromosome Deletion ; Chromosomes, Human, Y ; Humans ; Infertility, Male ; genetics ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; Varicocele ; genetics

OBJECTIVETo investigate the effect of high ligation of the spermatic vein (HLSV) on DNA fragmentation in varicocele (VC) patients.

METHODSThirty-four VC patients underwent HLSV. Sperm motion indexes and the results of papanicolaou staining and DNA fragmentation detection were analyzed before and 3 months after the operation according to the WHO guidelines.

RESULTSCompared with pre-operation, HLSV achieved a significant increase in the percentage of morphologically normal sperm (P < 0.01), and remarkable decreases in DNA fragmentation, sperm deformity index (SDI) and multiple anomalies index (MAI) (P < 0.01). The patients also showed significant increases in sperm concentration and the percentages of grade b sperm (P < 0.05) and grade a and a + b sperm (P < 0.01) after the operation. The post-operative percentages of sperm DNA fragmentation in those with grades I - III VC were markedly lower (P < 0.01), but showed no significant difference from that in those with subclinical VC (P > 0.05). The percentage of big-halo sperm was significantly increased (P < 0.01), while those of the medium-, small- and non-halo sperm remarkably decreased (P < 0.01) after HLSV.

CONCLUSIONHLSV can effectively improve the sperm quality of VC patients.

Adolescent ; Adult ; DNA Fragmentation ; Humans ; Ligation ; methods ; Male ; Sperm Count ; Sperm Motility ; Spermatozoa ; physiology ; Varicocele ; genetics ; surgery ; Veins ; surgery ; Young Adult

OBJECTIVETo investigate the hereditary tendency of varicocele.

METHODSWe included in this study 112 varicocele patients, 117 direct male relatives of the patients, and 100 healthy men as controls. We compared the incidence of varicocele tween the direct relative group and the control group.

RESULTSThe direct male relatives of the varicocele patients had a significantly higher incidence of varicocele than the healthy controls (36.8% vs 17%, P < 0.05).

CONCLUSIONThe increased incidence of varicocele in the direct male relatives of the patients indicated a hereditary tendency of the disease.

Adolescent ; Adult ; Case-Control Studies ; Humans ; Infertility, Male ; Male ; Middle Aged ; Pedigree ; Varicocele ; epidemiology ; genetics ; Young Adult

BACKGROUND: In addition to Klinefelter's syndrome, microdeletion of Yq is the most common genetic cause of male infertility; 15% of azoospermic or 5-10% of oligozoospermic males have Yq deletions. We evaluated a Yq microdeletion kit (LG Life Sciences, Korea) for identifying microdeletions in the azoospermic factor (AZF) regions of the Yq. METHODS: The kit was designed to amplify 3 regions of the AZF gene (AZFa, AZFb, and AZFc) using 15 sequence-tagged sites. We evaluated the preclinical performance of the kit. For clinical validation, 58 patients including 25 idiopathic azoospermic or oligozoospermic patients were examined. RESULTS: We observed clear bands on electrophoresis of DNA, up to a DNA concentration of 3.12 ng/microliter; the known microdeletion regions of all 6 reference cell-lines (Coriell, USA) were accurately detected and no false positive/negative results showed with normal female (n=11) and fertile male (n=15) specimens. This kit could identify the same microdeletions in the common regions, similar to another commercial kit. Among the 58 male infertile patients, 7 (12.1%) had microdeletions of the Yq. Among the idiopathic azoospermic (n=22) and oligozoospermic (n=3) patients, 3 (12.0%) had microdeletions. Further, 2 of 21 varicocele patients (9.5%), 1 of 4 patients with testicular failure, and 1 patient with a 45,X/46,XY mosaic had microdeletions. CONCLUSIONS: The kit was effective for detecting microdeletions of the Yq. We identified microdeletions in 12% of the infertile patients. This Y chromosome microdeletion detection kit is useful for screening Yq microdeletions in infertile patients.
Azoospermia/genetics ; *Chromosome Deletion ; *Chromosomes, Human, Y ; Electrophoresis, Agar Gel/methods ; Female ; Humans ; Infertility, Male/genetics ; Male ; Oligospermia/genetics ; Polymerase Chain Reaction/*methods ; Reagent Kits, Diagnostic ; Reproducibility of Results ; Seminal Plasma Proteins/*genetics ; Sensitivity and Specificity ; Varicocele/genetics

Glutathione S-transferases (GSTs), superoxide dismutase 2 (SOD2) and NAD(P)H:quinone oxidoreductase 1 (NQO1) are anti-oxidant enzyme genes. Polymorphisms of GSTs, SOD2 and NQO1 have been reported to influence individual susceptibility to various diseases. In an earlier study, we obtained preliminary findings that a subset of glutathione S-transferase T1 (GSTT1)-wt patients with varicocele may exhibit good response to varicocelectomy. In this study, we extended the earlier study to determine the distribution of genotype of each gene in the infertile population and to evaluate whether polymorphism of these genes affects the results of surgical treatment of varicocele. We analyzed 72 infertile varicocele patients, 202 infertile patients without varicocele and 101 male controls. Genotypes of GSTs were determined by polymerase chain reaction (PCR). Genotyping of SOD2 and NQO1 was performed using the PCR-restriction fragment length polymorphism (PCR-RFLP) method. A significantly better response to varicocelectomy was found in patients with the GSTT1-wt genotype (63.2%) and NQO1-Ser/Ser genotype (80.0%) than in those with GSTT1-null genotype (35.3%) and NQO1-Pro/Pro or NQO1-Pro/Ser genotype (45.2%), respectively. The frequencies of glutathione S-transferase M1/T1, SOD2 and NQO1 genotypes did not differ significantly among the varicocele patients, idiopathic infertile patients and male controls. GSTT1 genotype is associated with improvement of semen parameters after varicocelectomy. As the number of patients with NQO1-Ser/Ser genotype was not sufficient to reach definite conclusions, the association of NQO1 genotype with varicocelectomy requires further investigation.
Adult ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Infertility, Male ; etiology ; genetics ; surgery ; Male ; NAD(P)H Dehydrogenase (Quinone) ; genetics ; Polymorphism, Genetic ; Predictive Value of Tests ; Superoxide Dismutase ; genetics ; Treatment Outcome ; Varicocele ; complications ; genetics ; surgery

OBJECTIVETo investigate the effects of experimental left varicocele (ELV) on the expression of sperm associated antigen 11 (SPAG11) mRNA and its protein isomer SPAG11E in the testis and epididymis of adolescent rats, and to explore the mechanism of infertility caused by varicocele.

METHODSThe experimental left varicocele model was established in the adolescent male Sprague-Dawley rats. Two and 4 weeks after the operation, the changes of SPAG11 mRNA and SPAG11E expression in the testis and epididymis were detected using immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe expected product of SPAG11 367 bp amplified by RT-PCR was detected only in the epididymis. SPAG11E protein was observed mainly in the acrosomal vesicles and acrosome of round and elongating spermatids of the seminiferous epithelium, in the cytoplasm of Leydig cells, and in the supranuclear region of principle cells and stereocilia of the epididymal epithelium. Imaging and statistical analysis showed that SPAG11 mRNA and SPAG11E protein expressions in the left epididymis of the 2- and 4-week ELV groups presented a remarkable decrease (P < 0.05 or P < 0.01) compared with the right side and the corresponding control group, and the same decreased change in the left epididymis (P < 0.05 or P < 0.01) and an obvious reduction of SPAG11E immunopositive reaction in the right epididymis (P < 0.01) were noted in the 4-week group as compared with the 2-week group. No statistical difference of SPAG11E expression in the bilateral testes was found (P > 0.05) between the ELV group and the control, as well as between the 2- and 4-week ELV groups.

CONCLUSIONSPAG11 is a specific gene expressed in the epididymis. The localization and expression of SPAG11E exhibited a region- and cell-specific pattern in both the testis and epididymis of adolescent rats. The expression levels of both SPAG11 mRNA and SPAG11E protein altered obviously in ELV rats. The results suggest that SPAG11 may not only play an important role in spermatogenesis and sperm maturation, but also be associated with varicocele-induced male infertility or subfertility.

Animals ; Antigens, Surface ; genetics ; metabolism ; Disease Models, Animal ; Epididymis ; metabolism ; Glycopeptides ; genetics ; metabolism ; Immunohistochemistry ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Testis ; metabolism ; Time Factors ; Varicocele ; physiopathology

OBJECTIVETo study the influence of varicocele on sperm chromatin structure and sperm motility.

METHODSRoutine semen analysis and sperm chromatin structure assay (SCSA) were performed in a varicocele group (n=74) and a control group (n=89).

RESULTSSperm concentration (41.4 +/- 38.7] x 10(6)/ml) grade a+b sperm percentage ([31.7 +/- 16.9]% and sperm viability ([62.8 +/- 22.2]%) in the varicocele group were evidently lower than those ([80.9 +/- 63.1] x 10(6)/ml, [46.8 +/- 20.5]%, [77.2 +/- 17.5])% in the control group (P < 0.05) and so were VCL, VSL and VAP ([37.4 +/- 12.5 microm/s, [23.4 +/- 7.8] microm/s, [26.5 +/- 8.2] microm/s) in the varicocele group than those ([42.4 +/- 10.7] microm/s, [27.3 +/- 7.3] microm/s, [30.7 +/- 7.8] microm/s) in the control (P < 0.05). MAD was increased (P < 0.01), and the COMP alphat of SCSA (23.2 +/-16.2) was obviously higher in the former than in the latter (14.1 +/- 11.8) (P < 0.05).

CONCLUSIONVaricocele causes damage to sperm DNA and changes sperm motility, which may result in male infertility.

Adolescent ; Adult ; Chromatin ; genetics ; metabolism ; DNA Damage ; Humans ; Male ; Sperm Count ; Sperm Motility ; genetics ; physiology ; Spermatozoa ; cytology ; metabolism ; Varicocele ; genetics ; physiopathology