PubMed · 9634840
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A P Blanchard, L Hood. 1996. Sequence to array: probing the genome's secrets.. https://doi.org/10.1038/nbt1296-1649
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BACKGROUND: Standard sperm characteristics are poor predictors of the outcome of IVF treatments. On the contrary, sperm genome quality has been emphasized for several years as playing a major role in early embryogenesis, thus in the success of IVF attempt. METHODS: Sperm DNA fragmentation from a selected group of 104 couples undergoing assisted reproductive techniques (ART) (IVF: n = 50; and ICSI: n = 54) was measured by TUNEL assay and correlated with semen and ART outcomes. RESULTS: A negative correlation was found between sperm characteristics and the proportion of sperm showing DNA fragmentation. For fragmentation >10%, a significant decrease of the fertilization rate was observed. No correlation was found between sperm DNA fragmentation and embryo quality. A high proportion of sperm with fragmented DNA was a pejorative factor to obtain pregnancies when ICSI was performed, but there was no relationship when conventional IVF was performed. CONCLUSIONS: The proportion of sperm with DNA fragmentation appears to be potentially useful as a predictor of ICSI outcome, whereas embryo quality based on morphological criteria, appeared unaffected by DNA fragmentation.
BACKGROUND & OBJECTIVE: Gene therapy is the frontier of life science. There is no perfect method to evaluate gene expression without invasion at present. Medical imaging connecting with molecular biology might be helpful; however, the technology is just on the horizon. The authors conducted this study in vitro by transferring reporter tyrosinase gene into HepG2 cell to apply magnetic resonance imaging (MR) for evaluating gene expression. METHODS: The plasmid of pcDNA3tyr carried the full-length cDNA of tyrosinase gene was transfected into HepG2 cell by lipofectin, its property of synthesizing melanin was used to produce high signal in T1WI MR image and then to evaluate gene expression. Further identification were performed with searching melanin granules by Fontana staining and searching cDNA of tyrosinase gene using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: (1)The melanin granules were found in HepG2 cell using Fontana staining. (2)The cDNA fragment of tyrosinase gene was detectable in transferred HepG2 cell by RT-PCR. (3)Plasmid of pcDNA3tyr was transfected into HepG2 cell and synthesized a large amount of melanin in HepG2 cell; the synthetic melanin appeared high signal in T1WI MR imaging as same as natural melanin and was enough to be detected by MR. And further, the signal intensity was positively related to the amount of transferred plasmid. CONCLUSION: The fact that synthetic melanin of HepG2 cell can be detected by MR demonstrates that medical imaging connecting with molecular biology can be used to evaluate the result of gene expression in vitro.