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1.
Reprod Biomed Online ; 46(3): 483-490, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36642559

RESUMO

RESEARCH QUESTION: Non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) avoids the possible detrimental impact of invasive PGT-A on embryo development and clinical outcomes. Does cell-free DNA (cfDNA) from spent blastocyst culture medium (BCM) reflect embryonic chromosome status better than trophectoderm (TE) biopsy? DESIGN: In this study, 35 donated embryos were used for research and the BCM, TE biopsy, inner cell mass (ICM) and residual blastocyst (RB) were individually picked up from these embryos. Whole genome amplification (WGA) was performed and amplified DNA was subject to next-generation sequencing. Chromosome status concordance was compared among the groups of samples. RESULTS: The WGA success rates were 97.0% (TE biopsy), 100% (ICM), 97.0% (RB) and 88.6% (BCM). Using ICM as the gold standard, the chromosomal ploidy concordance rates for BCM, TE biopsy and RB were 58.33% (14/24), 68.75% (22/32) and 78.57% (22/28); the diagnostic concordance rates were 83.33% (20/24), 87.50% (28/32) and 92.86% (26/28); and the sex concordance rates were 92.31% (24/26), 100% (32/32) and 100% (28/28), respectively. Considering RB the gold standard, the chromosome ploidy concordance rates for BCM and TE biopsy were 61.90% (13/21) and 81.48% (22/27); the diagnostic concordance rates were 71.43% (15/21) and 88.89% (24/27); and the sex concordance rates were 91.30% (21/23) and 100% (27/27), respectively. CONCLUSIONS: The results of niPGT-A of cfDNA of spent BCM are comparable to those of invasive PGT-A of TE biopsies. Modifications of embryo culture conditions and testing methods will help reduce maternal DNA contamination and improve the reliability of niPGT-A.


Assuntos
Ácidos Nucleicos Livres , Diagnóstico Pré-Implantação , Gravidez , Feminino , Humanos , Diagnóstico Pré-Implantação/métodos , Reprodutibilidade dos Testes , Blastocisto/patologia , Aneuploidia , Testes Genéticos/métodos , Biópsia
2.
Reprod Biol ; 22(4): 100681, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35987159

RESUMO

Melatonin is a key neuroendocrine hormone that promotes spermatogenesis and sperm motility, but the underlying mechanisms remains poorly understood. In this study, we aimed to investigate the possible roles of m6A (N6--methyl-adenosine) in mediating melatonin-regulated spermatogonia activity alterations. In this study, mouse-derived GC-1 spermatogonia (spg) cell line was used as the in vitro cellular model. The viability, proliferation rates and apoptosis of spermatogonia were detected via CCK-8, Edu staining and flow cytometry respectively. Total m6A level was quantitated by dot blot, while mRNA and proteins contents in spermatogonia were measured by qRT-PCR and western blot respectively. Differentially expressed mRNAs were characterized by deep RNA sequencing method. Results showed that melatonin significantly promoted viability and proliferation rate while inhibited apoptosis in the GC-1 spg cells. The total m6A levels in GC-1 spg cells were also greatly increased by melatonin treatment, accompanied by remarkable expressional elevation of the m6A writer KIAA1429. Moreover, the regulation of GC-1 spg cell viability, proliferation and apoptosis by melatonin were greatly abrogated by KIAA1429 silencing but effectively strengthened by KIAA1429 overexpression. In addition, KIAA1429 overexpression regulates multiple biological process and signaling pathways in spermatogonia such as the PI3K/AKT signaling. The PI3K inhibitor LY294002 effectively mitigated the regulation of spermatogonia activity by KIAA1429 overexpression under melatonin treatment. Taken together, melatonin promotes spermatogonia activity via enhancing KIAA1429 expression and m6A RNA methylation to activate the downstream PI3K/AKT signaling pathway.


Assuntos
Adenosina , Melatonina , Proteínas de Ligação a RNA , Espermatogônias , Animais , Masculino , Camundongos , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Melatonina/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Fosfatidilinositol 3-Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais , Motilidade dos Espermatozoides , Espermatogônias/metabolismo , Proteínas de Ligação a RNA/metabolismo , Adenosina/análogos & derivados , Adenosina/metabolismo
3.
Dermatoendocrinol ; 9(1): e1348444, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29485142

RESUMO

Objective: This study was to verify if the skin cleanser could help decrease the infection ratio of Demodex Folliculorum in acne patients. Methods: 132 participants with mild to moderate vulgaris acne participated in this monocentric, prospective, double-blind study. Dermatologists grading and Standardized Skin Surface Biopsy were performed in baseline and after using cleanser only 7 d later. Results: There was no significant difference between the 2 times for each type of acne, but the number of Demodex Folliculorum was significantly decreased compared with baseline. There was no relationship between the number of Demodex Folliculorum and the total number of acne lesions. Limitations: Short follow-up time in 7 d. Conclusion: Using the cleanser could decrease the average number of Demodex Folliculorum in only 7 d in mild to moderate acne patients. There is no relationship between Demodex and acne lesions number.

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