Impact of cesarean scar defect on reproductive outcomes after assisted reproductive technology. / å‰–å®«äº§å­å®«ç˜¢ç—•ç¼ºæŸå¯¹è¾ åŠ©ç”Ÿæ®–å¦Šå¨ ç»“å±€çš„å½±å“åŠå¯¹ç­–.

In last twenty years, the cesarean section rate has been rising in China, leading to an increased prevalence of cesarean scar defect (CSD) and secondary infertility. CSD decreases receptivity of endometrium, induces adenomyosis and endometriosis, disrupts uterine micro-environment, causes difficulty of embryo transplant operation and further pregnancy complications. For women undergoing assisted reproductive technology (ART), CSD significantly reduces embryo implantation rate, clinical pregnancy rate and live birth rate. CSD can be effectively treated by hysteroscopic surgery, laparoscopic surgery and transvaginal surgery to increase the pregnancy rate. This article reviews the research progress on the impact of CSD on the reproductive outcomes of ART, the potential factors and related treatment, to provide reference for CSD management in ART patients.

RNA-Puzzles Round IV: 3D structure predictions of four ribozymes and two aptamers.

RNA-Puzzles is a collective endeavor dedicated to the advancement and improvement of RNA 3D structure prediction. With agreement from crystallographers, the RNA structures are predicted by various groups before the publication of the crystal structures. We now report the prediction of 3D structures for six RNA sequences: four nucleolytic ribozymes and two riboswitches. Systematic protocols for comparing models and crystal structures are described and analyzed. In these six puzzles, we discuss (i) the comparison between the automated web servers and human experts; (ii) the prediction of coaxial stacking; (iii) the prediction of structural details and ligand binding; (iv) the development of novel prediction methods; and (v) the potential improvements to be made. We show that correct prediction of coaxial stacking and tertiary contacts is essential for the prediction of RNA architecture, while ligand binding modes can only be predicted with low resolution and simultaneous prediction of RNA structure with accurate ligand binding still remains out of reach. All the predicted models are available for the future development of force field parameters and the improvement of comparison and assessment tools.

Modeling Loop Composition and Ion Concentration Effects in RNA Hairpin Folding Stability.

The ability to accurately predict RNA hairpin structure and stability for different loop sequences and salt conditions is important for understanding, modeling, and designing larger RNA folds. However, traditional RNA secondary structure models cannot treat loop-sequence and ionic effects on RNA hairpin folding. Here, we describe a general, three-dimensional (3D) conformation-based computational method for modeling salt concentration-dependent conformational distributions and the detailed 3D structures for a set of three RNA hairpins that contain a variable, 15-nucleotide loop sequence. For a given RNA sequence, the new, to our knowledge, method integrates a Vfold2D two-dimensional structure folding model with IsRNA coarse-grained molecular dynamics 3D folding simulations and Monte Carlo tightly bound ion estimations of ion-mediated electrostatic interactions. The model predicts free-energy landscapes for the different RNA hairpin-forming sequences with variable salt conditions. The theoretically predicted results agree with the experimental fluorescence measurements, validating the strategy. Furthermore, the theoretical model goes beyond the experimental results by enabling in-depth 3D structural analysis, revealing energetic mechanisms for the sequence- and salt-dependent folding stability. Although the computational framework presented here is developed for RNA hairpin systems, the general method may be applied to investigate other RNA systems, such as multiway junctions or pseudoknots in mixed metal ion solutions.

VfoldLA: A web server for loop assembly-based prediction of putative 3D RNA structures.

RNA three-dimensional (3D) structures are critical for RNA cellular functions. However, structure prediction for large and complex RNAs remains a challenge, which hampers our understanding of RNA structure-function relationship. We here report a new web server, the VfoldLA server (http://rna.physics.missouri.edu/vfoldLA), for the prediction of RNA 3D structures from nucleotide sequences and base-pair information (2D structure). This server is based on the recently developed VfoldLA, a model that classifies the single-stranded loops (junctions) into four different types and according to the loop-helix connections, assembles RNA 3D structures from the loop/junction templates. The VfoldLA web server provides a user-friendly online interface for a fully automated prediction of putative 3D RNA structures using VfoldLA. With a single-RNA or RNA-RNA complex sequence and 2D structure as input, the server generates structure(s) with the JSmol visualization along with a downloadable PDB file. The output result may serve as useful scaffolds for future structure refinement studies.

Predicting Cotranscriptional Folding Kinetics For Riboswitch.

On the basis of a helix-based transition rate model, we developed a new method for sampling cotranscriptional RNA conformational ensemble and the prediction of cotranscriptional folding kinetics. Applications to E. coli. SRP RNA and pbuE riboswitch indicate that the model may provide reliable predictions for the cotranscriptional folding pathways and population kinetics. For E. coli. SRP RNA, the predicted population kinetics and the folding pathway are consistent with the SHAPE profiles in the recent cotranscriptional SHAPE-seq experiments. For the pbuE riboswitch, the model predicts the transcriptional termination efficiency as a function of the force. The theoretical results show (a) a force-induced transition from the aptamer (antiterminator) to the terminator structure and (b) the different folding pathways for the riboswitch with and without the ligand (adenine). More specifically, without adenine, the aptamer structure emerges as a short-lived kinetic transient state instead of a thermodynamically stable intermediate state. Furthermore, from the predicted extension-time curves, the model identifies a series of conformational switches in the pulling process, where the predicted relative residence times for the different structures are in accordance with the experimental data. The model may provide a new tool for quantitative predictions of cotranscriptional folding kinetics, and results can offer useful insights into cotranscriptional folding-related RNA functions such as regulation of gene expression with riboswitches.

Predicting RNA Structure with Vfold.

In order to carry out biological functions, RNA molecules must fold into specific three-dimensional (3D) structures. Current experimental methods to determine RNA 3D structures are expensive and time consuming. With the recent advances in computational biology, RNA structure prediction is becoming increasingly reliable. This chapter describes a recently developed RNA structure prediction software, Vfold, a virtual bond-based RNA folding model. The main features of Vfold are the physics-based loop free energy calculations for various RNA structure motifs and a template-based assembly method for RNA 3D structure prediction. For illustration, we use the yybP-ykoY Orphan riboswitch as an example to show the implementation of the Vfold model in RNA structure prediction from the sequence.

Nanopore electric snapshots of an RNA tertiary folding pathway.

The chemical properties and biological mechanisms of RNAs are determined by their tertiary structures. Exploring the tertiary structure folding processes of RNA enables us to understand and control its biological functions. Here, we report a nanopore snapshot approach combined with coarse-grained molecular dynamics simulation and master equation analysis to elucidate the folding of an RNA pseudoknot structure. In this approach, single RNA molecules captured by the nanopore can freely fold from the unstructured state without constraint and can be programmed to terminate their folding process at different intermediates. By identifying the nanopore signatures and measuring their time-dependent populations, we can "visualize" a series of kinetically important intermediates, track the kinetics of their inter-conversions, and derive the RNA pseudoknot folding pathway. This approach can potentially be developed into a single-molecule toolbox to investigate the biophysical mechanisms of RNA folding and unfolding, its interactions with ligands, and its functions.