Effects of Low-Intensity Ultrasound on Cell Proliferation and Reproductivity

Ultrasound has been widely used in clinics. Cellular responses to low-intensity ultrasound are parameter-dependent. Proper parameter setting is vital to its exact use. To get guidelines for parameter setting, lowintensity ultrasound stimulation on the proliferation and reproductivity of Hep G2 and 3T3 cells in vitro was examined with a 1.06 MHz-generator by changing the parameters(including intensity, pulse repetition frequency and duty cycle)in a wide range. Cell viability and reproductivity at different time after sonication were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and colony formation assay to indicate timerelated proliferation. The results illustrate that ultrasound irradiation at 0.4—0.8 W/cm^2 and high pulse repetition frequency(100 Hz)can facilitate cell proliferation, while above 0.8 W/cm^2 would resist it. The extent of resistance closely correlated with duty cycle and pulse repetition frequency. Resistance effect at low pulse repetition frequency(1 Hz)is greater than that at high pulse repetition frequency(100 Hz)and not time-related. The influence of high pulse repetition frequency is time-accumulated, indicating cellular process involved. These findings would provide valuable guidelines for the application of low-intensity ultrasound in stem cell transformation and tissue engineering.

Temporal and Spatial Requirement of EMF1 Activity for Arabidopsis Vegetative and Reproductive Development

EMBRYONIC FLOWER （EMF） genes are required to maintain vegetative development via repression of flower homeotic genes in Arabidopsis. Removal of EMF gene function caused plants to flower upon germination, producing abnormal and sterile flowers. The pleiotropic effect of ernfl mutation suggests its requirement for gene programs involved in diverse developmental processes. Transgenic plants harboring EMF1 promoter：：glucuronidase （GUS） reporter gene were generated to investigate the temporal and spatial expression pattern of EMF1. These plants displayed differential GUS activity in vegetative and flower tissues, consistent with the role of EMF1 in regulating multiple gene programs. EMFI：：GUS expression pattern in emf mutants suggests organ-specific auto-regulation. Sense- and antisense （as） EMF1 cDNA were expressed under the control of stage- and tissue-specific promoters in transgenic plants. Characterization of these transgenic plants showed that EMF1 activity is required in meristematic as well as differentiating tissues to rescue emf mutant phenotype. Temporal removal or reduction of EMF1 activity in the embryo or shoot apex of wild-type seedlings was sufficient to cause early flowering and terminal flower formation in adult plants. Such reproductive cell memory is reflected in the flower MADS-box gene activity expressed prior to flowering in these early flowering plants. However, temporal removal of EMF1 activity in flower meristem did not affect flower development. Our results are consistent with EMF1＇s primary role in repressing flowering in order to allow for vegetative growth.

Advances of Long Noncoding RNAs-mediated Regulation in Reproduction

Objective： Advances in genomics and molecular biology have led to the discovery of a large group of uncharacterized long noncoding RNAs （lncRNAs）. Emerging evidence indicated that many lncRNAs function in multiple biological processes and its dysregulation otten causes diseases. Recent studies suggested that almost all regulatory lncRNAs interact with biological macromolecules such as DNA, RNA, and protein. LncRNAs regulate gene expression mainly on three levels, including epigenetic modification, transcription, and posttranscription, through DNA methylation, histone modification, and chromatin remodeling. LncRNAs can also affect the development of diseases and tllerefore be used to diagnose and treat diseases. With new sequencing and microarray techniques, hundreds oflncRNAs involved in reproductive disorders have been identified, but their functions in these disorders are undefined. Data Sources： This review was based on articles published in PubMed databases up to July 10, 2017, with the tbllowing keywords： ＂long noncoding RNAs＂, ＂＇LncRNA＂, ＂placentation＂, and ＂＇reproductive diseases＂. Study Selection： Original articles and reviews on the topics were selected. Results： LncRNAs widely participate in various physiological and pathological processes as a new class of important regulatory/＇,actors. In spermatogenesis, spermatocytes divide and differentiate into mature spermatozoa. The whole process is elaborately regulated by the expression of phase-specific genes that involve many strains of lncRNAs. Literature showed that IncRNA in reproductive cumulus cells may contribute to the regulation of oocyte maturation, fertilization, and embryo development. Conclusions： LncRNA has been found to play a role in the development of reproduction. Meanwhile, we reviewed the studies on how lncRNAs participate in reproductive disorders, which provides a basis fbr the study of [ncRNA in reproduction regulation.