A novel splicing mutation identified in a Chinese family with Alport syndrome and analysis of its pathogenicity / 医学研究生学报

Objective The purpose of this study was to identify a pathogenic variant in a Chinese family with Alport syndrome and analyze the pathogenicity of the variant. Methods Using targeted region capture and high-throughput sequencing technology, we identified the genetic variant of the proband with Alport syndrome, verified the variant in the family members by Sanger sequencing, and analyzed its influence on the pre-mRNA splicing process by in vitro minigene assay. Results A heterozygous variant c.2767G>T (p.Gly923Cys) was identified as a novel variant in exon 32 of the COL4A5 gene in the proband, which was confirmed by Sanger sequencing to be cosegregated with disease in the family. The minigene assay demonstrated that the c.2767G>T variant induced deletion of exon 32 of the COL4A5 gene. Conclusion A novel COL4A5 mutation was identified by targeted region capture and high-throughput sequencing, which has enriched the gene mutation spectrum of Alport syndrome. The exonic mutation c.2767G>T confirmed to be a splicing mutation by in vitro minigene assay, which may lead to a deeper insight into the molecular pathogenesis of Alport syndrome.

Detection of DPY19L2 gene mutation in a globozoospermia patient / 中华男科学杂志

<p><b>OBJECTIVE</b>Globozoospermia is mostly associated with homozygous deletion of the DPY19L2 gene. This study aimed to investigate the DPY19L2 gene mutation in a globozoospermia patient.</p><p><b>METHODS</b>We observed the sperm histomorphology of a patient with globozoospermia using Wright-Giemsa's staining and transmission electron microscopy, detected the mutation of the DPY19L2 gene by PCR amplification and DNA sequencing, and compared the findings with the sequences issued in the Genbank.</p><p><b>RESULTS</b>Wright-Giemsa's staining showed that all the spermatozoa were round-headed and lacked the acrosome, with the head nucleus darkly, fully and densely stained. Transmission electron microscopy revealed larger round sperm heads, with an even layer of unit membrane surrounding the nuclei and dispersed cytoplasmic vacuoles but no acrosomal structure. No DPY19L2 gene mutation was found by PCR amplification and DNA sequencing.</p><p><b>CONCLUSION</b>No homozygous mutation of the DPY19L2 gene was found in the globozoospermia patient, and therefore some other disease-causing genes might be involved.</p>

Protective effect of L-carnitine combined with sildenafil on the reproductive endocrine function of diabetic male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of L-carnitine (LC) combined with sildenafil on the reproductive endocrine function of male rats with diabetes mellitus (DM).</p><p><b>METHODS</b>A total of 40 male SD rats were randomly divided into five groups, group A taken as normal controls, and groups B, C, D and E made into DM models by injection of streptozotocin at 65 mg/kg. Then the rats in groups A and B were treated with normal saline, C with sildenafil at 5 mg per kg per d, D with LC at 300 mg per kg per d, and E with sildenafil at 5 mg per kg per d plus LC at 300 mg per kg per d, all via gastric gavage for 6 weeks, followed by determination of the levels of testosterone (T), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the serum of the rats.</p><p><b>RESULTS</b>After 6 weeks of treatment, the T, FSH and LH levels were (25.25 +/- 2.67) nmol/L, (5.78 +/- 0.61) IU/L and (625.21 +/- 43.45) ng/L in group A, (9.63 +/- 1.71) nmol/L, (1.98 +/- 0.42) IU/L and (479.89 +/- 27.62) ng/L in group B, (18.98 +/- 3.07) nmol/L, (5.08 +/- 0.33) IU/L and (586.57 +/- 31.72) ng/L in group C, (16.18 +/- 2.65) nmol/L, (4.63 +/- 0.30) IU/L and (540.78 +/- 25.52) ng/L in group D, and (23.65 +/- 2.66) nmol/L, (5.59 +/- 0.48) IU/L and (621.53 +/- 36. 40) ng/L in group E. The three parameters were significantly lower in B than in the other four groups (P < 0.01), and so were they in C and D than in A and E (P < 0.05), but showed no significant differences either between C and D (P > 0. 05) or between A and E (P > 0.05).</p><p><b>CONCLUSION</b>Six-week medication of either sildenafil or LC alone could increase the levels of T, FSH and LH in the serum of DM rats, but the combination of the two had an even more obvious increasing effect, which indicates a still better protective effect on the reproductive endocrine function of diabetic male rats.</p>

Meiotic analysis of spermatogenic cells in severe oligoasthenoteratozoospermia with chromosome 13 rearrangement / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the possible mechanisms of spermatogenic arrest in severe oligoasthenoteratozoospermia induced by supernumerary, ring-neocentric 13q12.3 --> 13q22 chromosome and reciprocal deletion.</p><p><b>METHODS</b>We performed a genomic-wide high-density oaCGH analysis for a case of oligoasthenoteratozoospermia with abnormal chromosome 13 to characterize the breakpoints of the chromosome involved or the gene deletion caused by the rearrangement. We also conducted a fluorescence in situ hybridization analysis on the germ cells using probes of 13q14/13qter to observe the pairing condition of homologous chromosome 13.</p><p><b>RESULTS</b>We identified by oaCGH analysis a microdeletion of 4 consecutive probes (A_16_P19757882, A_16_P02744617, A_14_ P108858 and A_16_P02744687 at chr13q12.3: 27979261 --> 28039191) with 59.93 kb between the FLT1 and POMP genes, with no annotated genes in the deleted region. The signals of 13q14 and 13qter were separated from each other in 90% of all the primary spermatocytes examined, indicating the unpairing of homologous chromosome 13 or synapse failure.</p><p><b>CONCLUSION</b>Chromosomal rearrangement-induced spermatogenesis failure is caused by the unpairing of the homologous chromosomes involved in the first meiotic division of germ cells.</p>

Cigarette smoking affects sperm plasma membrane integrity / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect sperm plasma membrane integrity (PMI) of cigarette smoking infertile males using SYBR-14/ PI fluorescent staining and flow cytometry and investigate its clinical significance.</p><p><b>METHODS</b>We collected semen samples from 132 cigarette smoking infertile men and 70 normal fertile controls, the former divided into a heavy-smoker group (> 20 cigarettes a day, n = 68) and a light-smoker group (< or = 20 cigarettes a day, n = 64). We performed computer-assisted semen analysis of the semen samples, and determined sperm PMI by flow cytometry after rinsing with PBS and staining by SYBR-14/PI, the sperm with normal PMI indicated as the percentage of those emitting green fluorescence (SYBR-14+/PI- %), dead sperm as the percentage of those emitting red (SYBR-14-/PI+), and moribund sperm as the percentage of those emitting both green and red (SYBR-14+/PI+).</p><p><b>RESULTS</b>Both the heavy- and light-smoker groups showed significant differences in SYBR-14-/PI+ % and SYBR-14+/PI- % from the normal controls (P < 0.01 or P < 0.05). SYBR-14+/PI- % was remarkably lower, while SYBR-14-/PI+ % markedly higher in the heavy-smoker than in the light-smoker group (P < 0.05). There was a significant correlation between SYBR-14+/PI- % and sperm motility (r = 0.938, P = 0.000).</p><p><b>CONCLUSION</b>SYBR-14/PI fluorescent staining and flow cytometry analysis could quickly and exactly detect sperm PMI. Cigarette smoking reduces sperm PMI and consequently sperm motility, which might be an important factor of male infertility.</p>

Application of Online Mendelian Inheritance in Man to medical genetics / 中华男科学杂志

Online Mendelian Inheritance in Man (OMIM, http://omim. org/) is a comprehensive, authoritative, practical and timely knowledgebase of human genes and genetic disorders. OMIM, as a genetic encyclopedia, provides an easy and straightforward access to information on human genetics to students, researchers and clinicians. This article presents an overview on the contents of OMIM and its application to medical genetics.

Ureaplasma urealyticum infection affects sperm plasma membrane integrity in infertile men / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the impact of Ureaplasma urealyticum (Uu) infection on the integrity of sperm plasma membrane in infertile males.</p><p><b>METHODS</b>Sixty-three semen samples were divided into a Uu infection group (n = 32) and a normal control group (n = 31). Conventional semen analyses were performed by computer-assisted semen analysis (CASA) and Uu detected by the culture method. The semen samples were washed with PBS and dyed by SYBR-14/PI double fluorescent staining, followed by detection of the integrity of sperm plasma membrane by flow cytometry. The percentage of the sperm with intact plasma membrane was indicated as the percentage of sperm emitting green fluorescence (SYBR-14+/PI-%).</p><p><b>RESULTS</b>The Uu infection group showed a significantly decreased integrity of sperm plasma membrane ([45.14 +/- 10.69]%) and reduced percentage of grade a + b sperm ([23.29 +/- 8.81]%) as compared with the normal control group ([72.68 +/- 9.91]% and [46.32 +/- 9.54]%) (P < 0.01). But there were no significant differences in the semen volume, pH value, and sperm concentration between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>Uu infection decreases the integrity of sperm plasma membrane, which might be an important factor of male infertility.</p>

Advances in rapid prenatal detection of fetal chromosome abnormalities / 中华男科学杂志

Rapid prenatal detection methods, including molecular cytogenetic analysis and ultrasonographic markers, are very important for prenatal diagnosis. The use of molecular cytogenetic techniques has significantly improved the rapid detection of aneuploidy and identification of small structural abnormalities of fetal chromosomes. At present, commonly used molecular cytogenetic techniques include fluorescence in situ hybridization (FISH), quantitative fluorescence PCR (QF-PCR), multiplex ligation-dependent probe amplification (MLPA) and microarray-based comparative genomic hybridization (array CGH). There is extensive evidence that major chromosomal abnormalities can be effectively detected by ultrasonography in the first and second trimesters of pregnancy. So we can combine molecular cytogenetic techniques with ultrasonographic markers to improve the identification of aneuploidies for chromosomes and the accuracy of prenatal diagnosis, and to reduce birth defects in newborns.

Effects of semen analysis on human sperm movement parameters at different times after semen collection / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of the computer-assisted semen analysis (CASA) on human sperm movement parameters at different times after semen collection.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>

Mutation of the USP26 gene in spermatogenesis dysfunction / 中华男科学杂志

The ubiquitin specific protease 26 (USP26) gene is located at Xq26.2 and present as a single exon on the X chromosome encoding for a protein of 913 amino acids. It belongs to a large family of deubiquitinating enzymes, and is exclusively expressed in the testis. There are conflicting reports on whether mutations in USP26 are associated with male infertility. This article updates the researches on the USP26 gene, its complicated relationship with male spermatogenesis dysfunction, the role of its mutation in male infertility, its geographical or ethnic distribution, and its evolution.

Mutation analysis of NF2 gene and clinical investigation in a Chinese family with neurofibromatosis type II / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Evaluation of sperm mitochondrial membrane potential in varicocele patients using JC-1 fluorescent staining / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect sperm mitochondrial membrane potential (MMP) of varicocele patients and investigate its clinical significance.</p><p><b>METHODS</b>Sixty-seven varicocele patients were divided into a VC1 (grade 1, n = 26), a VC2 (grade 2, n = 21) and a VC3 group (grade 3, n = 20). And 29 normal fertile volunteers were included in a control group ( m = 29). Conventional semen analyses were performed by computer-assisted semen analysis (CASA). Semen samples were washed, followed by JC-1 staining to evaluate the sperm MMP (JC-1+ %) by flow cytometry.</p><p><b>RESULTS</b>The sperm MMPs of the VC1, VC2 and VC3 groups were siginificantly lower ([56.29 +/- 16.32]%, P < 0.05; [45.04 +/- 13.21]%, P < 0.01; [31.63 +/- 12.91]%, P < 0.01) than that of the control ([76.21 +/- 13. 96]%). There was a significant positive correlation between the percentage of JC-1+ and that of grade (a + b) sperm (r =0.693, P=0.000).</p><p><b>CONCLUSION</b>The decreased MMP in the sperm of varicocele men might be one of the important causes of male infertility.</p>

Direct multiplex-PCR from whole blood for rapid detection of Y chromosome microdeletions / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a rapid and simple method to detect Y chromosome microdeletions directly using whole blood as starting material for multiplex-PCR.</p><p><b>METHODS</b>Using a self-prepared pHpH-Bufferq, multiplex-PCR amplification was directly carried out from the anticoagulant whole blood sample without DNA extraction step. Twelve sequence tagged sites (STS), namely SY84, SY86, SY127, SY134, SY124, SY132, SY152, SY157, SY239, SY242, SY254 and SY255, in AZFa, AZFb, and AZFc gene regions were detected in 5 different tubes. In order to ensure the validity of the experiments, sex-determining region Y (SRY) and X-linked or Y-linked zinc finger gene (ZFX/Y) were used as internal controls. Furthermore, conventional PCR using genomic DNA extracted from each blood sample was performed in parallel for evaluating the accuracy of the experiments.</p><p><b>RESULTS</b>A total of 156 male samples were detected, and a normal male sample and a female sample were used as a positive and a negative control respectively. The results showed that 144 samples had no deletion; one sample was AZF-deleted; one sample was AZFb-deleted; seven samples were AZFc-deleted; one sample was both AZFb- and AZFc- deleted; and two samples were all AZFa-, AZFb- and AZFc- deleted. The observed results from two kinds of starting material (whole blood and purified DNA) are completely consistent.</p><p><b>CONCLUSION</b>In our method, PCR amplification was directly carried out from whole blood without any DNA extraction step. So it has the advantages of low cost, simple process, time-saving operation and less cross-contamination. The whole process can be completed within 2 hours. Thus the efficiency of clinical detection is improved greatly.</p>

Determination of uric acid in the expressed prostatic secretion of chronic prostatitis patients and its clinical significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the level of uric acid (UA) in the expressed prostatic secretion (EPS) of chronic prostatitis patients and explore its clinical significance.</p><p><b>METHODS</b>A total of 91 patients with chronic prostatitis diagnosed by NIH standard were divided into a III A (n = 48) and a III B (n = 43) group, and healthy volunteers were selected as the control. The scores on the NIH-Chronic Prostatitis Symptom Index (CPSI) and the WBC count, pH value and UA level in EPS were evaluated for all the three groups.</p><p><b>RESULTS</b>The EPS UA concentration was (257.02 +/- 144.84) micromol/L in Group III B, significantly higher than in Group III A, (159. 73 +/- 121.49) micromol/L, (P < 0.01), and the control, (78.55 +/- 44.53) micromol/L, (P < 0.01). The level of EPS UA was correlated negatively with pH value (r = -0.398, P = 0.000), but positively with CPSI-P, CPSI-U and CPSI-T (r = 0.436, 0.316 and 0.403, P < 0.01).</p><p><b>CONCLUSION</b>Backflow of urine into prostatic ducts might cause chemical inflammation reaction by increasing UA concentration. There is a close relationship between the UA level in EPS and chronic prostatitis symptoms. Determination of the UA level in EPS is of great significance for the diagnosis and treatment of chronic prostatitis.</p>

AZF microdeletions are not related with recurrent spontaneous abortion / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the relationship between azoospermia factor (AZF) microdeletions of the Y-chromosome and recurrent spontaneous abortion.</p><p><b>METHODS</b>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.</p><p><b>RESULT</b>AZF microdeletions were found neither in the chorionic villus samples nor in the blood samples.</p><p><b>CONCLUSION</b>There is no relationship between AZF microdeletions and recurrent spontaneous abortion.</p>

Evaluation of CD4+ CD25+ regulatory T cells in the peripheral blood of recurrent spontaneous abortion patients / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the role of CD4+ CD25+ regulatory T cells (CD4+ CD25+ Tr) in the pathogenesis of recurrent spontaneous abortion.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>URSA might be associated with the decreased percentage of CD4+ CD25bright Tr, which plays an important role in fetomaternal immunologic tolerance.</p>

Follicle-stimulating hormone autoantibody is involved in idiopathic spermatogenic dysfunction / 亚洲男科学杂志(英文版)

<p><b>AIM</b>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.</p><p><b>METHODS</b>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).</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Genetic causes of male infertility / 中华男科学杂志

The causes of spermatogenetic failure found in most cases of non-ohstmctive azoospermia or severe oligospermia remain largely unclear. It is estimated that in about 30% of the cases, male infertility is due to genetic causes, including chromosomal abnormalities, Y chromosome microdeletions, gene mutations, etc. Klinefelter's syndrome and microdeletions in the Y chromosome long arm (Yq) represent the most frequent molecular genetic cause of severe infertility. Gene mutations involved in male infertility include the cystic fibrosis transmembrane conductance regulator (CFTR) gene, androgen receptor (AR) gene, insulin-like factor 3 (INSL3) gene and leucine-rich repeat-containing G-protein coupled receptor 8 (LGR8) gene. CFTR mutations cause cystic fibrosis, absence of vas deferens and non-obstructive azoospermia. The AR gene mutations are responsible for the androgen insensitivity syndrome and spermatogenetic damage. And INSL3 and LGR8 gene mutations have been associated with abnormalities in testis descent and cryptorchidism. Meta-analyses have revealed a significant association between the polymorphism and male infertility only for partial AZFc deletion, CAG repeat length in the AR gene and methylenetetrahydrofolate reductase (MTHFR) gene. This paper mainly reviews the genetic causes of male infertility and the genetic polymorphisms possibly associated with male infertility.

Observation of spermatogenic cells for infertile patients with Y-chromosomal microdeletion / 中华男科学杂志

<p><b>OBJECTIVE</b>To assess the spermatogenic function of the infertile patients with Y-chromosomal microdeletion.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Prenatal diagnosis of two pregnancies with risk of chromosomal disorders / 中华男科学杂志

<p><b>OBJECTIVE</b>To report the prenatal diagnosis of 2 cases of pregnancy with the risk of chromosomal disorders. In Case 1, the pregnant woman had a daughter with testicular regression syndrome and a segmental duplication of Ypter --> Yp11.2 and a deletion of Yq11.23 --> Yqter. In Case 2, both the pregnant woman and her husband were carriers of chromosomal balanced translocation.</p><p><b>METHODS</b>Two samples of amniotic fluid were obtained at the 19th week of gestation for fetal karyotype analysis. For Case 1, FISH with a probe of Xp/Yp subtelomere was performed on the metaphase of the amniotic fluid, genomic DNA of the amniotic fluid extracted and multiplex PCR conducted for AZF regions. Both the pregnant women underwent sonography to confirm the karyotypic diagnosis.</p><p><b>RESULTS</b>Cytogenetic, FISH and multiplex PCR analysis of the cultured amniotic fluid cells from Case 1 showed a normal male karyotype, and ultrasound scan of the fetus showed normal male external genitalia and normal development. Cytogenetic analysis of the cultured amniotic fluid cells from Case 2 revealed a karyotype of balanced translocation with t(13 ; 14) from the father, and no abnormality of the fetus was found by ultrasound scan.</p><p><b>CONCLUSION</b>It is helpful to perform cytogenetical and molecular prenatal diagnosis in combination with ultrasound scan for the fetus with the risk of chromosomal disorders and subsequently for genetic counseling.</p>