Objective To detect the sperm apoptosis rate and level of reactive oxygen species (ROS) in seminal plasma and explore their correlation among infertile males. Methods Ninety-two inferitile males were divided into varicocele (VC) group (n=32), leukocytospermia group(n=30) and the other cause group (n=30), and another 24 in vitro fertilization sperm samples were sereved as controls. The routine sperm parameters including seminal pH, sperm viability and sperm density were examined by computer assisted sperm analysis, the sperm apoptosis rate was asseseed using Annexin V/PI staining, and the ROS level in seminal plasma was detected by TBA method. The differences in seminal parameters between three infertile groups and control group were compared, and the correlation of sperm apoptosis rate with level of ROS in seminal plasma was explored in each group. Results The sperm viability of three infertile groups was significantly lower than that of control group (P<0.01). The sperm apoptosis rates and levels of ROS in seminal plasma in VC group and leukocytospermia group were significantly higher than those in control group (P < 0.05 or P < 0.01). The sperm apoptosis rate was positively correlated with the level of ROS in seminal plasma in leukocytospermia group(r=0. 573, P < 0.05). Conclusion The increased sperm apoptosis rate and level of seminal plasma ROS may be related to the infertility of patients with VC and leukocytospermia. The increased level of seminal plasma ROS may be one of the causes of increased sperm apoptosis rate in patients with leukocytospermia.

OBJECTIVETo investigate the variation of seminal plasma angiotension II (Ang II) in infertile men and its clinical implication.

METHODSAng II values in paired blood plasma and seminal plasma from 43 infertile men(13 azoospermia, 8 asthenozoopermia, 17 asthenozoospermia and 5 cases with normal semen parameters) and 10 normal controls were obtained by SPE-HPLC-RIA. All semen samples with spermatozoa were analyzed by CASA for sperm count, motility and other parameters. Acrosome reaction rate (AR) was assessed by triple-stain.

RESULTSThe mean concentration of seminal plasma Ang II was 4 times as high as that of blood plasma in all patients and controls (P < 0.01), but there was no correlation between them. The seminal plasma Ang II of azoospermic patients was higher than that of other infertile men and controls(P < 0.05), but no difference was found between the latter two groups. There was no correlation between seminal plasma Ang II values and other traditional parameters of sperm together with AR.

CONCLUSIONSSeminal plasma Ang II may be secreted locally in male reproductive tract. In addition to testis and epididymis, prostate and/or seminal vesicle may also be the source of it. The reason why seminal plasma Ang II of azoospermic patients is higher than that of others remains unknown. Further study is required to clarify the exact role of seminal plasma Ang II in the mechanisms of male fertility regulation.

Acrosome ; physiology ; Adult ; Angiotensin II ; analysis ; blood ; physiology ; Chromatography, High Pressure Liquid ; Humans ; Infertility, Male ; etiology ; metabolism ; Male ; Radioimmunoassay ; Renin-Angiotensin System ; physiology ; Semen ; chemistry

Objective To explore the impact factors for post-thaw embryo survival rate and clinical pregnancy rate in frozen-thawed embryo transfer program. Methods The clinical data of 573 cycles of frozen-thawed embryo transfers were retrospectively analysed. Groups were divided according to the pre-freeze embryo quality, pre-freeze embryonic developmental stage, frozen-thawed embryo quality and cryopreservation technique, respectively, and post-thaw embryo survival rates and/or clinical pregnancy rates were compared among groups. Results The clinical pregnancy rate of high quality pre-freeze embryo was significantly higher than that of low quality pre-freeze embryo (31.8% vs 20.0%) (P< 0.05). There was no significant difference in the post-thaw survival rates and clinical pregnancy rates between embryos frozen at day 2 of ferrtilization and those frozen at day 3 of ferrtilization(79. 1% vs 82.9% and 25.5% vs 31.2%, respectively) (P>0.05). The clinical pregnancy rates of the transfer cycles only with fully intact embryos and with mixed embryos were significantly higher than that only with partially damaged embryos(36.7% vs 24.1% and 29.2% vs 24.1%, respectively)(P<0.05). The post-thaw survival rate and post-thaw high-quality embryo rate were significantly higher in those processed with modified cryopreservation technique than in those processed with original cryopreservation technique (82.0% vs 66.3% and 50.0% vs 27.5%, respectively)(P<0.05). Conclusion Pre-freeze embryo quality, post-thaw embryo survival rate and post-thaw embryo quality have a positive correlation to subsequent clinical pregnancy rate. Favorable cryopreservation technique may ensure the success of post-thaw embryo recovery and transfer.

OBJECTIVESTo analyze human spermatozoa membrane proteins by two-dimensional gel electrophoresis and to provide a basis for drawing the protein map of normal human spermatozoa membrane proteins.

METHODSSpermatozoa were purified by Percoll density centrifugation, and spermatozoa membrane proteins were analyzed by two-dimensional gel electrophoresis using isoelectric focusing and polyacrylamide gel electrophoresis.

RESULTSAbout 800 protein spots could be identified by the imaging analysis system. The molecular weight and isoelectric point of most proteins were 20,000 to 100,000 and 3.0 to 7.0 respectively.

CONCLUSIONSThere are more than 800 types of proteins in the human spermatozoa membrane. The spermatozoa membrane protein can be identified with a certain precision by two-dimensional gel electrophoresis.

Electrophoresis, Gel, Two-Dimensional ; Humans ; Image Processing, Computer-Assisted ; Isoelectric Focusing ; Isoelectric Point ; Male ; Membrane Proteins ; chemistry ; Molecular Weight ; Spermatozoa ; chemistry

AIMTo study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism.

METHODSIn 18 healthy men, the seminal sperm motility and HSP90 levels were studied before and after cryopreservation using SDS-PAGE, Western blotting and computerized image analysis.

RESULTSThe sperm motility declined significantly after cryopreservation (P<0.01). The average grey level and the integrated grey level of sperm HSP90 before cooling were 34.1+/-3.2 and 243.0+/-21.6, respectively, while those after thawing were 23.2+/-2.5 and 105.7+/-28.5, respectively. Both parameters were decreased significantly (P<0.01). No HSP90 was found in the seminal plasma before and after cryopreservation.

CONCLUSIONHSP90 in human spermatozoa was decreased substantially after cryopreservation. This may result from protein degradation, rather than leakage into the seminal plasma.

Blotting, Western ; Cryopreservation ; HSP90 Heat-Shock Proteins ; metabolism ; Humans ; Male ; Semen Preservation ; Sperm Motility ; Spermatozoa ; cytology ; metabolism

OBJECTIVESTo investigate the role of Undecanoate (Andriol), as a kind of testosterone, in regulating the relaxation of isolated rat corpus cavernosum in vitro.

METHODSThe castrated rats were given high and low dosage Andriol respectively, compared with intact and castrated rats. After treatment of 4 weeks, the corpora cavernosa were cut, trimmed as to strips. Norepinephrine(NE) was added to contract each of the tissue strips. Next, sodium nitroprusside(SNP), electric functional stimulation(EFS) were used to relax the strips. Percent relaxations were examined.

RESULTSHigh-dose Andriol(20 mg/kg) was significantly effective to castrated rats on percent relaxation induced by 10(-3) mol/L SNP and EFS. Low-dose Andriol(10 mg/kg) was also effective to relax strips induced by 10(-3) mol/L SNP. According to statistics, the differences were significant (P < 0.01).

CONCLUSIONSThe percent relaxations of castrated rats were increased after taking Andriol, and it could increase the relaxation of corpara cavernosa.

Animals ; Castration ; Male ; Muscle Relaxation ; drug effects ; Muscle, Smooth ; drug effects ; physiology ; Penis ; cytology ; Rats ; Rats, Sprague-Dawley ; Testosterone ; analogs & derivatives ; pharmacology

OBJECTIVESTo detect the levels of reactive oxygen species (ROS), superoxide dismutase(SOD) and interleukin 8(IL-8) in seminal plasma of infertile patients, and evaluate the possible relationship between those levels.

METHODSSemen was collected from normal donors (15 cases), infertile men without infection (16 cases), and infertile men with infection (leukocytospermia, 11 cases). The routine analysis of semen was accomplished, and then the levels of IL-8, malondialdehyde (MDA), SOD, and white blood cell (WBC) were examined. The correlative analysis between the level of ROS and other parameters in these populations was made.

RESULTSIn leukocytospermic group, the levels of MDA, WBC, and IL-8 were higher than those in the other two groups (P < 0.001). Significantly positive correlation was observed between IL-8 and MDA (r = 0.852, P < 0.001) and between the levels of IL-8 and WBC.

CONCLUSIONSThese findings suggest that increased oxidative stress in patients with leukocytospermia may cause the increase of IL-8(r = 0.818, P < 0.01). The increased oxidative stress may be due to defect in ROS scavenging system.

Adult ; Cytokines ; blood ; Humans ; Infertility, Male ; blood ; complications ; Male ; Male Urogenital Diseases ; blood ; complications ; Reactive Oxygen Species ; blood ; Semen

OBJECTIVETo explore the effects of growth hormone( GH) supplementation on erectile function and expression of nNOS in the intracavernosal nerves in aging rats.

METHODSTwenty male Sprague-Dawley rats aged 18 months were randomly divided into Groups A and B, and ten 2-month-old male Sprague-Dawley rats included in Group C. 1 U/(kg x d) GH was given to Group A, and the same volume of saline to Groups B and C. After 8 weeks of treatment, evaluation was made of the erections induced by apomorphine (APO), maximal intracavernous pressure (ICP) induced by intracavernous papaverine injection and expression of nNOS in the intracavernosal nerves by streptavidin-peroxidase immunohistochemical techniques.

RESULTSAfter 8 weeks of treatment, the erection frequency, maximal ICP and expression of nNOS in the intracavernosal nerves of the rats in Groups A and C were significantly higher than those in Group B (P < 0.05 or P < 0.01).

CONCLUSIONGrowth hormone supplementation can improve the erectile function of aging rats, which may be attributed to the increase in the expression of nNOS in the intracavernosal nerves.

Animals ; Apomorphine ; pharmacology ; Growth Hormone ; pharmacology ; Immunohistochemistry ; Male ; Nitric Oxide Synthase Type I ; biosynthesis ; Papaverine ; pharmacology ; Penile Erection ; drug effects ; Penis ; enzymology ; innervation ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the methods and conditions for the isolation, purification and culture of human spermatogonial stem cells (SSCs) on the feeder layer cells of human embryonic fibroblasts (hEFs).

METHODSSSCs isolated and purified from normal human fetal testicular tissues by sequential two-step enzyme digestion and Percoll uncontinuous density gradient centrifugation were cultured on the feeder layer cells of hEFs isolated from 5-9 weeks old human embryos. The surface markers SSEA-1 and OCT4 of the SSCs were detected by immunohistochemistry; the alkaline phosphatase (AKP) activity of the SSC clones measured; and the expressions of the SSC-related genes determined by RT-PCR.

RESULTSSSCs survived, proliferated and formed colonies on the feeder layers, and the colonies were highly positive for SSEA-1 and OCT4, with strong AKP activity and high expressions of the SSC-related genes.

CONCLUSIONThe feeder layer of hEFs supports the growth of human spermatogonial stem cells.

Cell Culture Techniques ; methods ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Fibroblasts ; cytology ; Humans ; Male ; Spermatogonia ; cytology ; Stem Cells ; cytology

OBJECTIVE: The leukemia cells from patients with T-cell acute lymphoblastic leukemia (T-ALL) were inoculated into NCG mice to establish a stable human T-ALL leukemia animal model. METHODS: Leukemia cells from bone marrow of newly diagnosed T-ALL patients were isolated, and the leukemia cells were inoculated into NCG mice via tail vein. The proportion of hCD45 positive cells in peripheral blood of the mice was detected regularly by flow cytometry, and the infiltration of leukemia cells in bone marrow, liver, spleen and other organs of the mice was detected by pathology and immunohistochemistry. After the first generation mice model was successfully established, the spleen cells from the first generation mice were inoculated into the second generation mice, and after the second generation mice model was successfully established, the spleen cells from the second generation mice were further inoculated into the third generation mice, and the growth of leukemia cells in peripheral blood of the mice in each group was monitored by regular flow cytometry to evaluate the stability of this T-ALL leukemia animal model. RESULTS: On the 10th day after inoculation, hCD45⁺ leukemia cells could be successfully detected in the peripheral blood of the first generation mice, and the proportion of these cells was gradually increased. On average, the mice appeared listless 6 or 7 weeks after inoculation, and a large number of T lymphocyte leukemia cells were found in the peripheral blood and bone marrow smear of the mice. The spleen of the mice was obviously enlarged, and immunohistochemical examination showed that hCD3⁺ leukemia cells infiltrated into bone marrow, liver and spleen extensively. The second and third generation mice could stably develop leukemia, and the average survival time was 4-5 weeks. CONCLUSION Inoculating leukemia cells from bone marrow of patients with T-ALL into NCG mice via tail vein can successfully construct a patient-derived tumor xenografts (PDTX) model.
Humans ; Animals ; Mice ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; Heterografts ; Bone Marrow ; Disease Models, Animal ; T-Lymphocytes ; Mice, SCID