[Comparative analysis of two assaysin detection of sperm DNA fragmentation index, flow cytometry and AI-based fluorescence microscopy, based on AO staining: A multicentre study].

OBJECTIVE: To study the correlation, consistency, and variations between two assays of DNA fragmentation index based on acridine orange (AO) staining via AI-based fluorescence microscopy(AI-DFI), and flow cytometry (FCM-DFI) across multiple centers. METHODS: We selected 421 male patients from Nanjing Drum Tower hospital ( Hospital G) (226 cases), Eastern Theatre General Hospital (Hospital J) (89 cases) and Jiangsu Province Hospital (Hospital S) (106 cases) . Semen samples from each patient were analyzed for routine semen parameters and for DFI using both AI fluorescence microscopy and flow cytometry. We studied the two methods' stability as well as the correlation, consistency, and variation between the two methods' results in various centers. RESULTS: The two replicate studies' results of AI-DFI and the three centers' FCM-DFI for linear regression analysis indicated strong stability (R2>0.9).Overall(Group A), the AI-DFI results demonstrated good correlation and consistency with the FCM-DFI results of three centers (r>0.85；ICC>0.9).The semen specimens were categorized into two groups: normal specimen group (group B) and abnormal specimen group (group C) (including asthenozoospermia, oligospermia, and semen samples with high impurities).Group C's results showed a decline in correlation and consistency when compared to group A and group B, whereas group B's results showed a little rise in correlation and consistency when compared to group A. Although the consistency and correlation between the results of the two DFI testing methods in the three centers were good, there was still a significant difference between Groups A and C (P<0.05), and in Group B there was a significant difference between the two DFI testing methods only in Hospital G (p＝0.02), with no significant difference in Hospitals J and S (P> 0.05). CONCLUSION: The two detection methods exhibit good stability and correlation. However, significant differences are observed in the DFI detection methods in samples with abnormal semen parameters and high complexity. The main reason for these significant differences may lie in the variations in detection principles. Each detection method has its own advantages, allowing clinical or research settings to choose between them based on laboratory conditions or specific requirements.

[Effects of sperm DNA fragmentation index on semen parameters and pregnancy outcomes in intrauterine insemination].

Objective: To analyze the correlation of the sperm DNA fragmentation index (DFI) level with semen parameters and pregnancy outcomes of artificial insemination of the husband (AIH) in the cycle of intrauterine insemination (IUI). METHODS: We collected the clinical data on 777 cases of IUI, including female clinical indicators, male semen parameters, sperm DFI and pregnancy outcomes. According to the DFI level, we divided the patients into three groups: DFI < 15%, 15% ≤ DFI < 30% and DFI ≥ 30%. RESULTS: The sperm DFI level was significantly elevated with the increased age of the males (P = 0.002) and closely related to the total number of motile sperm (P = 0.002) and total sperm motility (P = 0.000) before treatment, as well as to sperm concentration (P = 0.000), total sperm motility (P = 0.001) and total number of progressively motile sperm (P = 0.000) after density gradient centrifugation. The rate of clinical pregnancy was decreased in the DFI ≥ 30% group. There were no statistically significant differences between sperm DFI and the rates of clinical pregnancy and abortion. CONCLUSIONS: Male age significantly affects the sperm DFI level. Sperm DFI is closely related to sperm motility and total number of progressively motile sperm, but not to the rates of clinical pregnancy and abortion in patients undergoing IUI. IUI can be used as an effective method of assisted reproduction for male infertility./.

Analysis of human sperm DNA fragmentation index (DFI) related factors: a report of 1010 subfertile men in China.

BACKGROUND: Many factors may lead to sperm DNA damage. However, it is little known that the correlations of sperm DNA damage with obesity-associated markers, and reproductive hormones and lipids levels in serum and seminal plasma. METHODS: In our prospective study, a total of 1 010 subfertile men, aged from 18 to 50 years old, were enrolled from August 2012 through June 2015. Their obesity-associated markers, semen parameters, sperm acrosomal enzyme activity, seminal plasma biochemical markers, and reproductive hormones and lipids levels in serum and seminal plasma were detected. Sperm DNA fragmentation index (DFI) was determined by sperm chromatin structure assay. The correlations between DFI and each of the above-mentioned variables were analyzed. RESULTS: Spearman correlation analysis showed that sperm DFI was positively related to age and abstinence time (P<0.001). Sperm DFI was also positively related to semen volume and percent of abnormal sperm head (P<0.001), while negatively related to sperm concentration, progressive motility (PR), sperm motility, total normal-progressively motile sperm count (TNPMS), percent of normal sperm morphology (NSM), percent of intact acrosome and acrosomal enzyme activity (P<0.001). Sperm DFI was positively related to seminal plasma zinc level (P<0.001) but unrelated to seminal plasma total α-glucotase, γ-glutamyl transpeptidase (GGT) and fructose levels. There was no any correlation between sperm DFI and obesity-associated markers such as body mass index (BMI), waist-to-hip ratio (WHR), waist circumference (WC) and waist-to-height ratio (WHtR), and serum lipids levels, but there was positive correlation between sperm DFI and seminal plasma triglyceride (TG) and total cholesterol (TC) levels (P<0.001). Sperm DFI was positively related to serum luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels and seminal plasma FSH and estradiol (E2) levels (P<0.001), but unrelated to serum and seminal plasma testosterone (T) levels. The multivariate regression analysis for the variables which were significantly correlated with sperm DFI in Spearman correlation analysis showed that age, semen volume, sperm concentration, progressive motility, TNPMS and intact acrosome were independently correlated with sperm DFI. CONCLUSIONS: There are many potential factors associated with sperm DFI, including age, abstinence time, spermatogenesis and maturation, seminal plasma lipids and reproductive hormones levels. However, the potential effects of seminal plasma lipids and reproductive hormones on sperm DNA damage need still to be demonstrated by the studies with scientific design and a large size of samples.

[Effect of secretin on the expression of cPLA2 and mPGEs-1 in mouse endometrial stromal cell during early pregnancy].

Secretin, a gastrointestinal peptide, has been found to be expressed in mouse endometrial stromal cells (mESCs) during early pregnancy. In order to further investigate the function of secretin during embryo implantation, the expression levels of secretin, secretin receptor, cytosolic phospholipase A2 (cPLA2) and membrane prostaglandin E synthase 1 (mPGEs-1) were detected in the mice uterus from day 4 to 8 of pregnancy by real-time PCR, ELISA and in situ hybridization. mESCs isolated and cultured from day 4 of pregnancy were transfected with secretin expression vectors or treated with H89, a PKA inhibitor. Then the expression levels of cPLA2, mPGEs-1 and cAMP responsive element-binding protein (CREB) were detected by real-time PCR and Western blot. The concentration of prostaglandin E2 (PGE2) in the supernatant was determined by ELISA. The result showed that secretin, cPLA2 and mPGEs-1 mRNA expression increased gradually in implantation sites from day 5 to day 7 of pregnancy with the same tendency. The secretin levels in serum were significantly higher on days 6, 7 and 8 of pregnancy than that on day 5 of pregnancy. The concentration of secretin was significantly higher in implantation sites on days 6, 7 than that in non-implantation site on day 5. Transfection of secretin expression vector promoted cPLA2, p-cPLA2 and mPGEs-1 expressions in mESCs, but not PGE2 level in the supernatant. H89 could effectively inhibit the expression of CREB, p-CREB, p-cPLA2 and cPLA2 induced by secretin. The results showed that the increased secretin expression in mESCs during embryo implantation may promote p-cPLA2, cPLA2 and mPGEs-1 expression, and the promotion may be through PKA signaling pathway.

[Protective effect of epigallocatechin gallate against sperm abnormality in mice].

OBJECTIVE: To investigate the protective effect of epigallocatechin gallate (EGCG) on mouse sperm in vivo. METHODS: A total of 64 six-week-old male Kuming mice were randomly divided into eight groups of equal number to be treated with normal saline (negative control), Cyclophosphamide (CP) at 30 mg/kg (positive control), and CP followed by EGCG (experimental) at 20, 40, and 80 mg/kg, respectively, given every other day for 10 days. At 4 and 5 weeks after treatment, the bilateral testes of the mice were harvested for examination of sperm abnormality. RESULTS: EGCG did not increase the rate of CP-induced sperm abnormality in the mice, but reduced it instead with the prolonged time of treatment. CONCLUSION: EGCG protects mouse sperm in vivo.

[Waist-hip ratio correlates with the levels of reproductive endocrine hormones in abdominally obese males].

OBJECTIVE: To study the impact of abdominal obesity on the production of male reproductive endocrine hormones. METHODS: This study included 342 male patients at the andrology clinic, aged 19 -47 years and higher than 160 cm. We measured their waistlines, hiplines and waist-hip ratio, detected the levels of serum estradiol (E2), testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH) and free testosterone (FT) by chemiluminescence and radioimmunoassay, and analyzed the correlation of the waist-hip ratio with the levels of reproductive endocrine hormones. Abdominal obesity was defined as the waist-hip ratio > 0.9. RESULTS: In the 342 male patients, there were 62 cases of abdominal obesity and 280 cases of the normal somatotype (waist-hip ratio < or = 0.9). The waist-hip ratio was negatively correlated with the T level (r = -0.163, P = 0.003) and the T/LH ratio (r = -0.13, P = 0.02). Both the T level and T/LH ratio were significantly reduced in the abdominal obesity patients ([14.51 +/- 4.53] nmol/L and 2.26 +/- 0.36) as compared with the normal somatotype controls ([17.21 +/- 4.23] nmol/L and 4.61 +/- 0.19) (P < 0.05). CONCLUSION: The waist-hip ratio has a significant negative correlation with the T level and T/LH ratio, and the serum T level is significantly lower in men with abdominal obesity than in those of the normal somatotype.

[Effect of annexin 5 on StAR and testosterone synthesis related enzyme in rat Leydig cells].

OBJECTIVE: To investigate the effect of annexin 5 on the expressions at mRNA levels and protein levels of StAR, P450scc, 3ß-HSD, 17α-hydroxylase and 17ß-HSD in rat Leydig cells. METHODS: The primary rat Leydig cells were cultured for 24 h and then stimulated with 10(-9) mol/L annexin 5 for 12 h and 24 h respectively. Cellular total RNA and total protein were extracted respectively. The expressions of StAR, P450scc, 3ß-HSD, and 17α-hydroxylase and 17ß-HSD(10) mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR)and the protein levels were detected by Western blotting. RESULTS: Compared with the control group, at the mRNA level, after being treated with annexin 5 for 12 h, only 17ß-HSD(10) expression had a 26% increase (P<0.05) while the others had no significant difference. The expressions of StAR, P450scc and 3ß-HSD elevated 55%, 69% and 59%(P<0.05) respectively, and 17ß-HSD(10) increased 104%(P<0.01) while 17α-hydroxylase had no significant difference after being treated with annexin 5 for 24 h. At the protein level, after being treated with annexin 5 for 12 h, 17ß-HSD expression had a 39% increase (P<0.05). After 24 h, P450scc, 3ß-HSD and 17ß-HSD elevated 35%, 88% (P<0.05) and 47% (P<0.01) respectively while StAR had no significant difference. CONCLUSION: Annexin 5 regulates testosterone synthesis by affecting the expressions of P450scc, 3ß-HSD and 17ß-HSD at gene and protein levels.

[Mutation analysis of NF2 gene and clinical investigation in a Chinese family with neurofibromatosis type II].

OBJECTIVE: To report a heterozygous RNA-splicing mutation (IVS3+ 3A to C) of NF2 gene in a Chinese family with autosomal dominant neurofibromatosis type II and investigate the relationship between the genotype and phenotype. METHODS: The proband with bilateral vestibular schwannomas underwent gamma knife radiosurgery two years earlier. DNA of blood samples from all affected individuals, suspected individuals and unaffected relatives of the family was extracted and amplified to detect the polymorphisms at loci D22S1150 and D22S268 that are linked with the NF2 gene. Two-point LOD score was calculated. The promoter region, 17 exons and exon/intron boundaries of NF2 gene were amplified and sequenced for the proband. The exon 3/intron 3 boundaries of NF2 gene was amplified and sequenced for the other 3 patients, 1 suspected individual, 9 unaffected members of the family and 150 unrelated controls. RESULTS: The result of two-point linkage analysis suggested that NF2 gene was a candidate gene (Zmax= 2.109, θ = 0.00, locus D22S1150). DNA sequencing revealed a heterozygous splicing mutation in intron 3 (IVS3+ 3A to C) for the proband. Identical mutation was also observed in the other 3 patients and 1 suspected individual. No mutation was found in the 9 normal family members and 150 unrelated controls, which was consistent with the clinical diagnosis. CONCLUSION: This is the first report of familial neurofibromatosis type II with a splicing mutation of IVS3+ 3A to C of the NF2 gene. The mutation might be responsible for the neurofibromatosis type II in the family.

[Effects of semen analysis on human sperm movement parameters at different times after semen collection].

OBJECTIVE: To investigate the effects of the computer-assisted semen analysis (CASA) on human sperm movement parameters at different times after semen collection. METHODS: Ninety-two semen samples with sperm density > or = 20 x 10(6)/ml and sperm liquefaction time < 20 min were placed in a incubation box at the temperature of 37 degrees C. Then the seminal parameters were analyzed with the computer-assisted semen analysis (CASA) system at 20, 30, 60 and 90 min after semen collection. RESULTS: The percentages of grade a and b sperm were significantly lower at 30, 60 and 90 min than at 20 min (P < 0.05), so were the percentages of grade c sperm at 60 and 90 min than at 20 and 30 min (P < 0.05), but there were no significant differences in the percentage of grade c sperm between the 20-min and 30-min groups (P > 0.05). The percentages of grade a + b and a + b + c sperm were also significantly lower at 30, 60 and 90 min than at 20 min (P < 0.05). The beat cross frequency (BCF) was significantly higher at 30 min than at 20 min (P < 0.05), while the lateral head amplitude (ALH) significantly lower at 90 min than at 30 min (P < 0.05). The sperm wobbliness (WOB) was significantly higher while the curvilinear velocity (VCL) significantly lower at 90 min than at 20 and 30 min (P < 0.05). Straightness (STR) at 30, 60 and 90 min, and average path velocity (VAP) and straight line velocity (VSL) at 90 min were significantly lower than at 20 min (P < 0.05). There were no significant differences in sperm density, average motion degree (MAD) and linearity (LIN) among the four groups (P > 0.05). CONCLUSION: The interval between semen collection and sperm routine analysis needs to be standardized. The results of this study suggest that sperm movement parameters of normal liquefied semen samples are relatively constant at 30 -60 min after semen collection.

[HONO Observation and Assessment of the Effects of Atmospheric Oxidation Capacity in Changzhou During the Springtime of 2017].

HONO measurement was conducted using a wet-chemistry-based method at the Changzhou Environmental Monitoring Center in April 2017. HONO ranged from 0.2-13.9 µg·m-3 with an average of (2.9±2.3) µg·m-3. O3, HCHO, volatile organic compounds, photolysis frequency, and meteorological parameters were simultaneously monitored.·OH concentration was simulated by a Master Chemical Mechanism box model and the daytime maximum·OH concentration ranged from 1.0×106 to 14×106 molecules per cubic centimeter. The formation rates of·OH by photolysis of HONO, O3, HCHO, H2O2, and alkene ozonolysis were calculated as well. The effects of the five sources on atmospheric oxidation capacity were revealed:O3 photolysis (46.4%) > HONO photolysis (41.1%) > alkene ozonolysis (10.9%) > HCHO photolysis (1.5%) > H2O2 photolysis (0.1%). HONO photolysis for OH radical production played a major role in the early morning, before with an increase in O3 concentration, O3 photolysis began to account for most of the·OH production. After 17:00, due to a significant decrease in the intensity of solar radiation, the alkene ozonolysis started playing a major role in the formation of·OH. The photolysis of formaldehyde and hydrogen peroxide played a negligible role in·OH radical production in this study.

Follicle-stimulating hormone autoantibody is involved in idiopathic spermatogenic dysfunction.

AIM: To detect the anti-follicle-stimulating hormone (FSH) antibody in idiopathic infertile patients and fertile subjects in order to determine the role of this antibody in patients with spermatogenic dysfunction. METHODS: The anti-FSH antibody in serum was detected by an enzyme-linked immunosorbent assay (ELISA). The functional and structural integrity of the sperm membrane was evaluated with hypo-osmotic swelling (HOS) test and the ultrastructure of the spermatozoa was investigated by transmission electron microscopy (TEM). RESULTS: The extent of positive FSH antibody in the patients with oligozoospermia and/or asthenozoospermia was significantly higher than that in the fertile subjects and infertile patients with normal sperm concentration and motility, but it was significantly lower than that in the patients with azoospermia. The extent of anti-FSH antibody in the patients with azoospermia was significantly greater than that in patients with oligospermia and/or asthenospermia, infertile people with normal sperm density and motility and fertile people. The hypo-osmotic swelling test showed that the percentage of HOS-positive spermatozoa (swollen) was 45.1 mu 3.5% in the FSH antibody-positive group and 59.1% micro 6.2% in the FSH antibody-negative control group. The percentage of functional membrane damage to spermatozoa was significantly higher in the anti-FSH antibody-positive group than in the control group. TEM showed that the outer acrosomal membrane was located far from the nucleus, and detachment of the acrosome was found in the FSH autoantibody-positive group. CONCLUSION: These data suggest that the presence of anti-FSH antibody is strongly correlated with the sperm quantity and quality in idiopathic male infertility. Anti-FSH antibody may be an important factor causing spermatogenic dysfunction and infertility.

[Ureaplasma urealyticum infection in the genital tract reduces seminal quality in infertile men].

OBJECTIVE: To understand Ureaplasma urealyticum (Uu) infection, analyzed the influence of Uu infection on the seminal quality and the accessory genetical gland function in male infertility patients, and investigate its mechanism. METHODS: We cultured 202 semen samples collected from male infertility patients and analyzed the influence of Uu infection on seminal parameters and the biochemical indexes of the seminal plasma. RESULTS: The Uu infection rate was 33.7% in the infertile males, with no statistic differences between the Uu positive and negative groups either in the average age (28.9 +/- 4.7 yrs vs 29.6 +/- 4.0 yrs, P = 0.250) or in the seminal quantity (2.93 +/- 1.32 ml vs 2.86 +/- 1.52 ml, P = 0.774). The sperm density, motility and vitality were (84.37 +/- 52.92) x 10(6) ml, (44.62 +/-22.13) % and (38.40 +/- 15.61) % in the Uu positive group, significantly lower than (101.90 +/- 43.90) x 10(6) ml, (51.83 +/- 19.88) % and (44.45 +/- 15.47) % in the Uu negative group (P = 0.025, P = 0.036 and P = 0.020). The seminal pH value was normal in both of the groups, but significantly higher in the Uu positive than in the negative group (7.32 +/- 0.10 vs 7.19 +/- 0.29, P = 0.003). VCL, VSL, VAP and MAD were significantly lower, while BCF was significant higher in the former than in the latter [(33.97 +/- 8.96) microm/s vs (39.70 +/- 8.14) microm/s, t = 4.113, P < 0.001; (22.29 +/- 6.06) microm/s vs (25.20 +/- 6.67) microm/s, t = 2.684, P = 0.008; (25.96 +/- 6.83) microm/s vs (30.02 +/- 6.81) microm/s, t = 3.537, P < 0.001; 46.60 +/- 13.68 vs 54.23 +/- 15.14, t = 3.112, P = 0.002; (6.12 +/- 1.89) Hz vs (5.22 +/- 1.64) Hz, t = 3. 164, P = 0.002]. All the five indexes were influenced by Uu infection. Compared with the negative group, the seminal plasma alpha-glucosidase was significantly decreased in the positive group [(40.0 +/-18.7) U/ml vs (47.9 +/- 21.0) U/ml, t = 2.248, P = 0.026], and the risk of the decrease was 2.12 times higher. No statistic difference was observed in seminal plasma acid phosphatase and seminal plasma fructose between the two groups. CONCLUSION: Uu infection in the genital tract is an important factor of seminal quality reduction in infertile men and may cause a decreased secretion of alpha-glucosidase in the epididymis, but it hardly influences the prostate and seminal vesicle.

[Comparison of the parameters obtained by sperm quality analyzer V and computer-aided sperm analysis system].

OBJECTIVE: To assess the differences between the main parameters obtained by sperm quality analyzer V (SQA-V) and computer-aided sperm analysis system (CASA), and to investigate their application to sperm quality analysis for fertile and infertile men. METHODS: Twelve fresh semen samples from fertile volunteers and 73 from infertility patients were detected with SQA-V and CASA for sperm concentration and motility, the percentage and concentration of motile sperm, sperm motility index (SMI), amplitude of lateral head displacement (ALH), beat cross frequency (BCF), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN = VSL/VCL) and straightness (STR = VSL/VAP). The correlation between the parameters obtained by the two devices were analyzed. RESULTS: Significant differences were observed in the above parameters between the fertile and infertile groups. An obvious consistency was noted between the results from SQA-V and those from CASA in sperm concentration (r = 0.58, P < 0.01), motile sperm concentration (r = 0.75, P < 0.01) and average sperm velocity (r = 0.59, P < 0.01). Significant correlations were found between the SMI from SQA-V and STR, LIN, BCF, VCL, VSL and VAP from CASA (P < 0.05). CONCLUSION: There is a consistency between the results from SQA-V and those from CASA. Both the devices can detect the seminal differences between different cohorts of patients.

[AZF microdeletions are not related with recurrent spontaneous abortion].

OBJECTIVE: To study the relationship between azoospermia factor (AZF) microdeletions of the Y-chromosome and recurrent spontaneous abortion. METHODS: We collected 26 chorionic villus samples from abortive male embryos and 51 blood samples from the husbands whose wives had recurrent spontaneous abortion, extracted genomic DNA from the samples and detected 12 STSs in the AZFa, AZFb and AZFc regions of Yq11.2 by amplification multiplex PCR. RESULT: AZF microdeletions were found neither in the chorionic villus samples nor in the blood samples. CONCLUSION: There is no relationship between AZF microdeletions and recurrent spontaneous abortion.

[Evaluation of CD4+ CD25+ regulatory T cells in the peripheral blood of recurrent spontaneous abortion patients].

OBJECTIVE: To explore the role of CD4+ CD25+ regulatory T cells (CD4+ CD25+ Tr) in the pathogenesis of recurrent spontaneous abortion. METHODS: Peripheral blood samples were collected from 29 women with unexplainable recurrent spontaneous abortion (the URSA group) and another 20 with normal pregnancy (the control group). The percentage of CD4+ CD25+ Tr in the peripheral blood was measured by flow cytometry. RESULTS: The rate of CD4+ CD25bright Tr in the URSA patients ([1.98 +/- 0.96]%) was significantly lower than that in the control group ([3.21 +/- 1.25]%, P < 0.05), while the percentages of CD4+ CD25+ and CD4+ CD25dim and the ratio of CD4+ CD25bright/CD4+ were not significantly different between the two groups (P > 0.05). CONCLUSION: URSA might be associated with the decreased percentage of CD4+ CD25bright Tr, which plays an important role in fetomaternal immunologic tolerance.

[Determination of plasma homocysteine in oligospermia and/or asthenospermia patients].

OBJECTIVE: To detect the level of fasting plasma homocysteine (Hcy) in patients with oligospermia and/or asthenospermia and to investigate its clinical significance. METHODS: Semen quality analyses and fasting plasma Hcy determination were performed for 86 infertility patients (21 with oligospermia, 32 with asthenospermia and 33 with oligo-asthenospermia) and 19 normal fertile volunteers. The results were compared. RESULTS: The level of plasma Hcy was significantly higher in the infertility patients than in the normal controls (P < 0.05) and negatively correlated with sperm concentration (r = -0.433, P < 0.01), the percentage of grade a sperm (r = -0.303, P < 0.05) and the percentage of grade a+b sperm (r = -0.339, P < 0.01). CONCLUSION: The increased level of human plasma Hcy directly or indirectly affects spermatogenesis and correlates negatively with oligospermia and/or asthenospermia.

[Observation of spermatogenic cells for infertile patients with Y-chromosomal microdeletion].

OBJECTIVE: To assess the spermatogenic function of the infertile patients with Y-chromosomal microdeletion. METHODS: Thirty-five 23-44 years old patients with microdeletions of Y chromosome were included in this study. Three semen analyses confirmed that 26 cases were non-obstructive azoospermia and 9 oligospermia with sperm count < 1 x 10(6)/ml. They were divided into 3 groups by the locus of deletion, 5 cases of AZFa + b + c deletion in group 1, 4 cases of AZFb + c and 3 cases of AZFb deletion in group 2, and 23 cases of AZFc deletion in group 3. Semen was collected and centrifuged, the supernatant removed and the centrifugate applied on the clean slides after dilution. Following Wright's-Giemsa staining, the slides were viewed under the microscope. Testis histopathological biopsy was performed for 6 of the cases. RESULTS: In group 1, no spermatogenic cells were observed but only Sertoli cells in 1 case, with a consistency between the result of spermatogenic cell test and that of testis biopsy. In group 2, spermatogenic cell tests revealed spermatocytes in 6 cases, 2 were proved by testis biopsy with sperm maturation arrest in the primary spermatocyte stage, and spermatogenic cells of all developmental stages were seen in 1 AZFb deletion patient with the same sperm maturation arrest as the above two. In group 3, only primary spermatocytes were detected by spermatogenic cell test in 5 oligospermia patients but no spermatogenic cells in the 15 azoospermia cases, and biopsy revealed 2 cases of Sertoli cell-only syndrome. CONCLUSION: The spermatogenic cell test can effectively assess the spermatogenic function of AZF deletion patients. Non-invasive and easily accepted by patients, it is highly recommendable for the evaluation of spermatogenesis of patients with Y-chromosomal microdeletion.

[A preliminary study of pre-analysis quality control for semen analysis].

OBJECTIVE: To discuss how some pre-analysis processes influence the results of semen analysis and how to minimize their influence on the accuracy of laboratory results based on the concept of total quality management (TQM). METHODS: We conducted semen quality analyses for 21 male volunteers, who had abstained from tobacco and alcohol for over 72 days for the purpose of fertilization, before and after the abstinence, and obtained their seminal parameters at 0.5, 1, 2 and 3 hours after semen sample collection. RESULTS: Sperm concentration, sperm motility and the percentage of grade a + b sperm were significantly higher after the abstinence of tobacco and alcohol than before (P < 0.01). With the lengthening of post-ejaculation time, there was a significant decrease in sperm motility and the percentage of grade a + b sperm (P < 0.05), but not in sperm concentration (P > 0.05). CONCLUSION: A lot of factors may affect the results of semen analysis, including the subjects' habits of drinking and smoking and the length of time after semen collection. Therefore, every procedure of semen analysis has to be dealt with very carefully so as to meet the requirements of TQM and achieve most reliable results for clinical use.

[Semen biochemical markers and their significance in the patients with premature ejaculation].

OBJECTIVE: To detect the changes of biochemical markers in the semen of premature ejaculation patients and investigate the correlation of the markers with premature ejaculation. METHODS: Fifty-six premature ejaculation patients and 60 males with normal sexual behavior were enrolled in this experiment. Acid phosphatase, alpha- glucosidase and fructose were assayed by the methods of glucose oxidase, disodium phenyl phosphate and disodium phenyl phosphate respectively. RESULTS: The contents of acid phosphatase, alpha-glucosidase and fructose were (36.37 +/- 31.33) U/ml, (39.97 +/- 22. 09) U/ml and (3.40 +/- 1.92) mg/ml in the premature ejaculation patients and (54. 27 +/- 20. 96) U/ml, (55.71 +/- 16.19) U/ml and (2.55 +/- 1.12) mg/ml in the normal control, respectively, with significant differences in the former two markers between the two groups. The rate of the abnormal content of both acid phosphatase and alpha- glucosidase was 31% and 13% (P < 0.05) , while that of the normal content of the three markers was 10% and 33% in premature ejaculation group and the control, respectively (P < 0. 05 ). CONCLUSION: The abnormality of both acid phosphatase and alpha-glucosidase is one of the causes of premature ejaculation. Because acid phosphatase and alpha- glucosidase reflect the functions of the prostate and epididymis, we should pay attention to the status of these two organs in the treatment of premature ejaculation.