Protocol for the derivation and culture of murine trophoblast organoids for CRISPR-Cas9 screening.

Murine trophoblast organoids present a more balanced array of trophoblast subtypes, rendering them a suitable platform for CRISPR-Cas9-based screening. Here, we present a protocol for the derivation and culture of murine trophoblast organoids from trophoblast stem cells or placentae. We describe steps for establishing and differentiating murine trophoblast organoids, the characterization of trophoblast organoids in both conditions, the generation of focused single guide RNA (sgRNA) libraries, and the subsequent screening using those libraries in murine trophoblast organoids.

Clinical Outcomes of Descemet Membrane Endothelial Keratoplasty for Iridocorneal Endothelial Syndrome With a Glaucoma Drainage Device.

Purpose: The purpose of this study was to evaluate the feasibility and clinical outcomes of Descemet membrane endothelial keratoplasty (DMEK) for treating iridocorneal endothelial (ICE) syndrome with a glaucoma drainage device (GDD).

Methods: In this retrospective, interventional case series, data of ICE eyes with a GDD treated with DMEK were collected at the Zhongshan Ophthalmic Center. A total of 24 patients (24 eyes) with mild-to-moderate ocular anterior segment anomalies together with good intraocular pressure (IOP) control preoperatively were included between March 10, 2014, and November 11, 2021.

Murine trophoblast organoids as a model for trophoblast development and CRISPR-Cas9 screening.

The placenta becomes one of the most diversified organs during placental mammal radiation. The main in vitro model for studying mouse trophoblast development is the 2D differentiation model of trophoblast stem cells, which is highly skewed to certain lineages and thus hampers systematic screens. Here, we established culture conditions for the establishment, maintenance, and differentiation of murine trophoblast organoids.

Chemical-induced phase transition and global conformational reorganization of chromatin.

Chemicals or drugs can accumulate within biomolecular condensates formed through phase separation in cells. Here, we use super-resolution imaging to search for chemicals that induce phase transition within chromatin at the microscale. This microscopic screening approach reveals that adriamycin (doxorubicin) - a widely used anticancer drug that is known to interact with chromatin - specifically induces visible local condensation and global conformational change of chromatin in cancer and primary cells.

Quercetin suppresses retinoblastoma cell proliferation and invasion and facilitates oxidative stress-induced apoptosis through the miR-137/FNDC5 axis.

Retinoblastoma (RB) constitutes a prevalent malignancy in clinic and usually occurs in children under the age of 5 years old. The increased frequency of malignant tumor metastases and the delayed diagnosis and treatment caused unsatisfactory therapeutic efficiency. Quercetin was formerly identified to impede tumor growth in certain malignancies.

DeepFundus: A flow-cytometry-like image quality classifier for boosting the whole life cycle of medical artificial intelligence.

Medical artificial intelligence (AI) has been moving from the research phase to clinical implementation. However, most AI-based models are mainly built using high-quality images preprocessed in the laboratory, which is not representative of real-world settings. This dataset bias proves a major driver of AI system dysfunction.

Transcription-coupled donor DNA expression increases homologous recombination for efficient genome editing.

Genomes can be edited by homologous recombination stimulated by CRISPR/Cas9 [clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated peptide 9]-induced DNA double-strand breaks. However, this approach is inefficient for inserting or deleting long fragments in mammalian cells. Here, we describe a simple genome-editing method, termed transcription-coupled Cas9-mediated editing (TEd), that can achieve higher efficiencies than canonical Cas9-mediated editing (CEd) in deleting genomic fragments, inserting/replacing large DNA fragments and introducing point mutations into mammalian cell lines.

Complete sequence of a retinopathy inbred C57BL/6 mouse model mitochondrial genome.

In the present work we undertook the complete mitochondrial genome sequencing of an important retinopathy model inbred C57BL/6 strain for the first time. Its mitogenome was 16,312 bp and coding 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes. A total of 96 SNPs were examined when compared to reference BN sequence.

[Clinical study in combining application of mitomycin and amnion transplantation together with pterygium resection].

Purpose: The aim of this study was to evaluate the effectiveness and safety of combining application of mitomycin and/or amnion transplantation when carrying out pterygium resection.

Methods: Pterygium resection, mitomycin application and amniotic membrane transplantation were carried out in three combinations: Pterygium resection plus local application of mitomycin C (MMC group, n=114), Pterygium resection plus amnion transplantation (AMT group, n=119) and Pterygium resection plus mitomycin C application and amniotic membrane transplantation (combined group, n=127). The patients were followed up for 6-18 months.