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1.
Journal of Southern Medical University ; (12): 1059-1064, 2019.
Article in Chinese | WPRIM | ID: wpr-773493

ABSTRACT

OBJECTIVE@#To explore the effect of small interfering RNA (siRNA)-mediated CEP55 gene silencing on the proliferation of mouse spermatogonia.@*METHODS@#Six patients with azoospermia diagnosed to have maturation arrest (3 cases) or normal spermatogenesis (3 cases) based on testicular biopsy between January 1 and December 31, 2017 in our center were examined for differential proteins in the testicular tissue using isobaric tags for relative and absolute quantitation (iTRAQ), and CEP55 was found to differentially expressed between the two groups of patients. We constructed a CEP55 siRNA for transfection in mouse spermatogonia and examined the inhibitory effects on CEP55 expressions using Western blotting and qPCR. The effect of CEP55 gene silencing on the proliferation of mouse spermatogonia was evaluated with CCK8 assay.@*RESULTS@#In the testicular tissues from the 6 patients with azoospermia, iTRAQ combined with LC/MS/MS analysis identified over two hundred differentially expressed proteins, among which CEP55 showed the most significant differential expression between the patients with maturation arrest and those with normal spermatogenesis. The cell transfection experiment showed that compared with the cells transfected with the vehicle or the negative control sequence, the mouse spermatogonia transfected with CEP55 siRNA showed significantly lowered expressions of CEP55 mRNA and protein ( < 0.05) and significantly decreased proliferation rate as shown by CCK8 assay ( < 0.05).@*CONCLUSIONS@#CEP55 may play a key role in spermatogenesis and may serve as a potential therapeutic target for non-obstructive azoospermia with maturation arrest.


Subject(s)
Animals , Humans , Male , Mice , Azoospermia , Genetics , Cell Cycle Proteins , Genetics , Gene Silencing , Nuclear Proteins , Genetics , Spermatogenesis , Spermatogonia , Tandem Mass Spectrometry , Transfection
2.
Journal of Southern Medical University ; (12): 1469-1474, 2014.
Article in Chinese | WPRIM | ID: wpr-329265

ABSTRACT

<p><b>OBJECTIVE</b>To assess the value of combined evaluation of serum follicle-stimulating hormone (FSH), inhibin B (INHB), chromosome karyotyping and AZF microdeletion of Y-chromosome (AZF-MD-Ych) in predicting the success of testicular sperm aspiration (TESA) in azoospermic patients.</p><p><b>METHODS</b>A total of 262 azoospermic patients were divided into two groups with normal (n=162) and abnormal (n=100) serum FSH levels. INHB levels, INHB/FSH ratio, chromosome karyotype patterns of the peripheral lymphocytes, and AZF-MD-Ych were compared between the two groups. Among the patients receiving TESA, the success rate of the procedure was compared between the two groups after excluding abnormalities in INHB, chromosome karyotype and AZF-MD-Ych.</p><p><b>RESULTS</b>Significant differences were found between the two groups in serum INHB level, INHB/FSH and chromosome karyotypes (P<0.05), but not in AZF-MD-Ych (P>0.05). After excluding the abnormalities in chromosome karyotypes, AZF-MD-Ych and INHB, sperms were obtained successfully by TESA from 61.82% (34/55) of patients with normal FSH but from none of those with abnormal FSH (P<0.01).</p><p><b>CONCLUSION</b>A combined evaluation of serum FSH, INHB, chromosome karyotypes and AZF-MD-Ych can effectively predict the success of TESA in azoospermic patients, and abnormalities in all the 4 indices suggest a very low success rate of sperm retrieval by TESA.</p>


Subject(s)
Humans , Male , Azoospermia , Chromosome Deletion , Chromosomes, Human, Y , Follicle Stimulating Hormone , Blood , Infertility, Male , Inhibins , Blood , Karyotyping , Prognosis , Sex Chromosome Aberrations , Sex Chromosome Disorders of Sex Development , Sperm Retrieval , Spermatozoa , Testis , Treatment Outcome
3.
Journal of Southern Medical University ; (12): 584-587, 2014.
Article in Chinese | WPRIM | ID: wpr-249402

ABSTRACT

<p><b>OBJECTIVE</b>This study is in an attempt to evaluate the diagnostic significance to predict the spermatogenesis of azoospermic men in examination of serum follicle-stimulating hormone (FSH) combination with serum inhibin B (INHB).</p><p><b>METHODS</b>Quantitative examination of serum FSH and INHB was performed in 95 case of azoospermic men. According to their classifications of testicular biopsy with histopathological examination, there were 20 patients of Sertoli cell only, 25 of hypospermatogenesis, 18 of spermatogenic maturation arrest (complete or incomplete), and 32 of normal spermatogenesis. The association of serum FSH and INHB levels with histopathological classifications were analyzed by using statistical software.</p><p><b>RESULTS</b>Serum FSH, INHB and INHB/FSH levels of Sertoli cell only differed with statistical significance from hypospermatogenesis, spermatogenic maturation arrest and normal spermatogenesis (P<0.05). FSH, in which there were no statistical significance among the latter three classifications (P>0.05). Serum FSH, INHB and INHB/FSH levels were no relationship with maturation arrest (P>0.05), but were negatively related to the other classifications (P<0.05). INHB level less than 28.55 pg/ml predicted Sertoli cell only in a sensitivity of 97% and a specificity of 85%.</p><p><b>CONCLUSION</b>Serum FSH and INHB levels is ineffective to distinguish the spermatogenic classifications from azoospermic men, but they are available to confirm the disease of Sertoli cell only. The other abnormalities of azoospermic men is also dependent on bioptic histopathology to confirm the subtypes.</p>


Subject(s)
Adolescent , Adult , Humans , Male , Middle Aged , Young Adult , Azoospermia , Blood , Diagnosis , Follicle Stimulating Hormone , Blood , Infertility, Male , Blood , Diagnosis , Inhibins , Blood , Oligospermia , Spermatogenesis , Testis , Physiology
4.
Journal of Southern Medical University ; (12): 857-861, 2014.
Article in Chinese | WPRIM | ID: wpr-249344

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the impact of spermatozoa from different sources on normal fertilization of oocytes, embryo quality and embryo developmental potential in intracytoplasmic sperm injection (ICSI) cycles.</p><p><b>METHODS</b>A retrospective analysis was conducted among 197 patients undergoing ICSI cycles in our center. The patients were classified into 3 groups according to the sources of semen, namely ejaculated spermatozoa group (n=102), percutaneous epididymal sperm aspiration (PESA) group (n=68), and testicular sperm aspiration (TESA) group (n=27). The ejaculated spermatozoa group was further classified into oligoasthenoteratozoospermia (n=67) and cryptozoospermia (n=35) subgroups. The normal fertilization, high-quality embryo, implantation and clinical pregnancy rates were compared among the groups; the rate of high-quality blastocyst formation in in-vitro culture of non-top quality embryos was also observed.</p><p><b>RESULTS</b>The patients with PESA showed significantly higher normal fertilization rate (75.6%) than those in oligoasthenoteratozoospermia (64.8%), cryptozoospermia (62.1%), and TESA (61.6%) groups (P<0.05). No significant differences were found in the high-quality embryo, implantation, and clinical pregnancy rates among the groups (P>0.05). The rate of high-quality blastocyst formation in the in-vitro culture of non-top quality embryos was also comparable among the groups (P>0.05).</p><p><b>CONCLUSION</b>Although spermatozoa obtained with by PESA is associated with a higher normal fertilization rate, the sources of spermatozoa do not significantly affect the embryonic quality and developmental potential in ICSI cycles.</p>


Subject(s)
Female , Humans , Male , Pregnancy , Asthenozoospermia , Embryo Implantation , Embryonic Development , Fertilization , Fertilization in Vitro , Oligospermia , Oocytes , Pregnancy Rate , Retrospective Studies , Semen , Sperm Injections, Intracytoplasmic , Sperm Retrieval , Spermatozoa
5.
Journal of Southern Medical University ; (12): 448-450, 2013.
Article in Chinese | WPRIM | ID: wpr-322026

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the association of sperm mobility parameters assessed by computer-assisted sperm analysis (CASA) with the rates of normal fertilization, oocyte cleavage and excellent embryos in in vitro fertilization (IVF) cycles.</p><p><b>METHODS</b>A total of 288 infertile women undergoing IVF cycles patients were divided into two groups according to the normal fertilization rate (≥50% and <50%), cleavage rate (≥90% and <90%), or excellent embryo rates (≥50% and <50%). The means of the sperm motility parameters analyzed by CASA twice before oocyte retrieval were recorded and analyzed using t-test in relation to the rates of normal fertilization, cleavage and excellent embryos in IVF cycles.</p><p><b>RESULTS</b>The mean curvilinear velocity (VCL), average path velocity (VAP), and amplitude of lateral head displacement (ALH) were significantly higher in women with a normal fertilization rate of ≥50% than in those with a normal fertilization rate of <50% (P<0.05). Women with an oocyte cleavage rate of ≥90% had significantly higher VCL and VAP than those with a cleavage rate of <90% (P<0.05). The VCL, straight line velocity (VSL), VAP, linearity, straightness, wobble coefficient, ALH, or beat-cross frequency showed no significant differences between women with excellent embryo rates of ≥50% and <50% (P>0.05).</p><p><b>CONCLUSION</b>The sperm motility parameters assessed using CASA are associated with normal fertilization and oocyte cleavage rates but not with excellent embryo rate in IVF cycles.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Fertilization in Vitro , Methods , Infertility, Female , Therapeutics , Semen Analysis , Sperm Motility , Therapy, Computer-Assisted
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