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OBJECTIVE@#To explore the correlation between microRNA (miRNA) differential expression and quality of embryo.@*METHODS@#The miRNA expression profiles of 8 blastocysts were detected by a TaqMan microRNA array, and miRNAs with a stable expression were selected. Additional blastocysts were selected, and the candidate miRNA was detected by real-time PCR. Meanwhile, chromosomal abnormalities of the embryos were detected by using next-generation sequencing, and the results were compared.@*RESULTS@#The expression of mir-720, mir-372, mir-886-3p and mir-512-3p was higher than that of miR-145, which suggested that mir-720, mir-372, mir-886-3p and mir-512-3p are related to early embryo development. The expression of miR-145 and mir-886-3p were significantly lower in the normal chromosome group. With the threshold values of above 9 and 3 for the relative expression of miR-145 and mir-886-3p, respectively, there was no embryo without a chromosomal abnormality.@*CONCLUSION@#There is a correlation between the expression level of specific miRNA and chromosomal abnormalities of embryos, which may be used as a novel biomarker for embryo selection.
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Multiple morphological abnormalities of sperm flagella (MMAF) is a type of teratospermia caused by genetic defects. The sperm motility is low due to absence of flagella, shortness, curling, bending or irregularity of sperms, and combination of various abnormalities. Ultrastructure may show flagellum assembly abnormalities, which are mainly manifested by the absence of microtubules in the axoneme and defects of various structures such as fibrous sheath, outer dense fiber, mitochondrial sheath and dynein arms. MMAF males are unable to reproduce naturally and require assisted reproductive technology to obtain offsprings. For the heterogeneity of molecular etiology of MMAF, the outcome of assisted reproduction may be different. Here the candidate genes of MMAF and their functional mechanisms are summarized, which may provide a reference for clinical diagnosis, treatment and research for this disorder.
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Objective To evaluate the value of sperm acrosomal arginine enzyme activity in the diagnosis and treatment of male infertility by studying its relationship with sperm motility and morphology.Methods The sperm acrosomal arginine enzyme activity was detected by chemical colorimetric method,and the routine parameters of sperm and sperm morphology were detected by computer-aided analysis.Results There were significant differences on sperm concentration,motility and progressive motility between normal sperm acroso-mal arginine enzyme activity group and abnormal sperm acrosomal arginine enzyme activity group(P<0.05). The percentage of deformities in head,neck and middle segment were significant different between normal sperm acrosomal arginine enzyme activity group and abnormal sperm acrosomal arginine enzyme activity group (P<0.05).Conclusion T he activity of sperm acrosin is a very efficient marker in sperm quality,and an effec-tive indicator of the evaluation of sperm fertilization potential and the diagnosis of male infertility.
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<p><b>OBJECTIVE</b>To detect FOXL2 gene mutation in a Chinese pedigree affected with blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) type I, and to explore its genotype-phenotype correlation.</p><p><b>METHODS</b>Peripheral blood samples were obtained from 3 patients and 19 healthy members from the pedigree for the isolation of genomic DNA. All exons and flanking sequences of the FOXL2 gene were amplified by PCR with 7 pairs of overlapping primers and sequenced.</p><p><b>RESULTS</b>DNA sequencing indicated that the BPES phenotype in this pedigree was caused by a hotspot c.843_859dup17 mutation. The same mutation was not found among the healthy members of the pedigree.</p><p><b>CONCLUSION</b>The c.843_859dup17 frameshift mutation probably underlies the BPES type I in this Chinese pedigree, which may manifest as either BEPS type I or type II.</p>
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Objective To investigate the transrectal ultrasonographic (TRUS) features of midline prostatic cysts and the semen analysis in infertile men. Methods The ultrasonographic characteristics of midline prostatic cysts were retrospectively analyzed in 34 infertile men, who underwent transrectal ultrasound and semen analysis in Suzhou Hospitial Affiliated to Nanjing Medical University from November 2013 to October 2014. Results Thirty-four patients were detected by transrectal ultrasound with anechoic areas in their prostates. Among them, the cysts could be divided into 3 groups:17 ejaculatory duct cyst, 11 mullerian duct cyst and 6 true prostatic cyst. Ejaculatory duct cysts was connected with ipsilateral seminal vesicle and pointed to seminal hillock showing a water-drop shape on longitudinal view of ultrasound. Mullerian dust cysts were located at the base of the prostates behind the urethra with a water-drop shape on longitudinal view. True prostatic cysts were inside the prostates or under the prostate capsules with a round or oval shape either on longitudinal view or transverse view of ultrasound. Therefore, these midline prostate cysts could be differentiated by their anatomical position and shape. These patients then underwent semen analysis. By semen analysis, there were 13, 4, 1 aspermia and 2, 6, 2 oligospermia in patients with ejaculatory dust cysts, mullerian duct cysts, and true prostatic cysts, respectively. Conclusions Transrectal ultrasound can provide accurate information about the anatomical position and shape of midline prostatic cysts in infertile men. Combined with semen analysis, transrectal ultrasound can provide a reliable evidence for clinical treatment.
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Objective To construct survivin-targeting siRNA-expressing plasmid.Methods DNA sequence correspond to siRNA targeting survivin was designed and synthesized,and cloned into plasmid pRNAT-U6.1/Neo to produce surviving-targeting plasmid.Two oligos in the template with cohesive BamHⅠ and HindⅢ sites were prepared and annealled to form the insert fragment for siRNA vector.The vector was cut with BamHⅠ and HindⅢ and ligated with the insert fragment using T4 ligase.The recombinant vector was confirmed by restriction digestion and DNA sequencing,and then was transfected into T24 cells with Lipofectamine TM2000 and the expression of survivin was detected by real-time quantitive PCR.Results DNA sequencing for the PCR product showed that the recombinant vector pRNAT-U6.1/Neo-survivin was successfully constructed without any base pair mutation.The plasmid pRNAT-U6.1/Neo-survivin could efficiently reduce the expression of survivin and confer G-418 resistance in T24 cells.Conclusion The siRNA-expressing plasmid which were successfully constructed and transfected into T24 cells in this study may facilitate the application of RNA interference technique,and lay foundation for further studies on the function of survivin.