MiR-181a-5p may regulate cell proliferation and autophagy in myopia and the associated retinopathy.

The mechanism of myopia and the associated retinopathy remains unclear, and dysregulated microRNAs (miRNAs) are implicated in this disease. In this research, we purposed to find out the regulatory function that miRNAs play in myopia and the associated retinopathy. We first performed miRNA microarray analysis in a lens-induced myopia mouse model and found that miR-9-5p, miR-96-5p, miR-182-5p, miR-183-5p, and miR-181a-5p were elevated in the myopic retina. Then, we examined the functions and regulatory mechanisms of miR-181a-5p utilizing the human retinal pigment epithelium (RPE) cell line ARPE-19 by overexpressing miR-181a-5p. RNA sequencing (RNA-Seq) and qRT-PCR analysis were employed to identify differentially expressed genes after transfection. The qRT‒PCR outcomes, immunoblotting, and immunofluorescence indicated that the SGSH expression was significantly hindered through miR-181a-5p overexpression. MiR-181a-5p overexpression has the ability to elevate RPE cell proliferation and induce autophagy by targeting SGSH. We validated the negative influence of miR-181a-5p on the SGSH expression through luciferase reporter assays, which demonstrated its ability to target the 3' untranslated region of SGSH. The reversal of implications of miR-181a-5p overexpression was achieved through SGSH upregulation. We provided novel perspectives into the miR-181a-5p function in regulating myopia development and may serve as a target for therapy and molecular biomarker for myopia.

A robust localization approach for Percutaneous Nephrolithotomy (PCNL): A single-center retrospective study.

BACKGROUND: Percutaneous Nephrolithotomy (PCNL) represents the gold standard treatment method for cases with large kidney stones. As a critical step in performing PCNL, the procedure of establishing a safe and accurate nephrostomy tract will dramatically impact the treatment quality of patients with large-sized kidney stones. OBJECTIVE: This work attempts to describe a new and improved process of establishing an accurate nephrostomy tract and clinically evaluate the effectiveness and safeness of this proposed methodology. METHODS: This work represents a retrospective single-center study carried out between August 2013 and November 2019. The collected samples consist of 937 patients who were operated on using PCNL coupled with our proposed procedure. Briefly, a preoperative B-ultrasonography was firstly performed to decide the puncture point in a simulated surgical position where was marked with ureteral catheter segments (2-3 cm). A computed tomography (CT) scan was followed to correct the anchor points in the simulated surgical position. After this, an accurate puncture operation was performed under the real-time guidance of intraoperative B ultrasound. RESULTS: Examining this study, 851 subjects with renal stones and 86 subjects with ureteropelvic junction stones were included for the PCNL operation project. All samples were grouped with Guy's grading system: grade I, II, III, and IV patients there were 0.00%, 42.69%, 51.01%, and 6.30%, respectively. Among these patients, the average age was 48.49 ± 10.80 years old, with a male to female ratio of around 1.73:1. CONCLUSIONS: This study showed that our developed method warrants an accurate and safe PCNL operation that involves the process of establishing the nephrostomy tract. Other advantageous attributes of this new PCNL process include negligible radiation exposure, lesser complications, and low failure rates. More importantly, this new localization approach is particularly attractive for hospitals that are new to the field of adopting PCNL considering its safeness, effectiveness, and learnability.

Identification of a novel missense mutation of MIP in a Chinese family with congenital cataracts by target region capture sequencing.

Congenital cataract is both clinically diverse and genetically heterogeneous. To investigate the underlying genetic defect in three-generations of a Chinese family with autosomal dominant congenital cataracts, we recruited family members who underwent comprehensive ophthalmic examinations. A heterozygous missense mutation c.634G > C (p.G212R) substitution was identified in the MIP gene through target region capture sequencing. The prediction results of PolyPhen-2 and SIFT indicated that this mutation was likely to damage the structure and function of MIP. Confocal microscopy images showed that the intensity of the green fluorescent signal revealed much weaker signal from the mutant compared to the wild-type MIP. The expressed G212R-MIP was diminished and almost exclusively cytoplasmic in the HeLa cells; whereas the WT-MIP was stable dispersed throughout the cytoplasm, and it appeared to be in the membrane structure. Western blot analysis indicated that the protein expression level of the mutant form of MIP was remarkably reduced compared with that of the wild type, however, the mRNA levels of the wild-type and mutant cells were comparable. In conclusion, our study presented genetic and functional evidence for a novel MIP mutation of G212R, which leads to congenital progressive cortical punctate with or without Y suture.

A Head-Mounted Spectacle Frame for the Study of Mouse Lens-Induced Myopia.

The mouse model has been widely employed to explore the mysteries of myopia. For now, existing techniques for induction of experimental myopia in mice can be classified into three types: (1) devices directly glued to the fur; (2) devices attached using a combination of glue and sutures; (3) devices attached using a skull-mounted apparatus. These techniques each have its advantages, disadvantages when considering the devices stability, safety, complexity, effectiveness, and so forth. Thus, techniques for myopia induction in mice have yet to be further refined to popularize the applications. In this pilot study, we introduce a new head fixation device named the head-mounted spectacle frame apparatus for the study of mouse lens-induced myopia. Surgical procedures for device attachment were relatively simple and easy to learn in our study. Effective myopia induction was validated by retinoscopy refraction and axial length measurement using optical coherence tomography. In addition, it showed improved compliance and reliable safety when compared to the published methods. The head-mounted spectacle frame apparatus provides a new choice for the study of lens-induced myopia in mouse. It also allows for the use of form deprivation, making it attractive for future experimental mouse myopia trials.

Topical Use of NaCl Solution with Different Concentration Affects Lens Transparency in Anesthetized Mice.

PURPOSE: To study the influence of NaCl solution with different concentration on lens transparency in anesthetized mice. METHODS: Four kinds of NaCl solution with different concentration were prepared as eye drops to imply graded osmolarity (100, 300, 500 and 1000 mOsmol/kg). Five groups of anesthetized mice were set-up to induce lens opacity, in which four groups were treated with NaCl solution and another group naturally exposed to air. The lens opacity was graded as no opacity, mild, medium and severe opacity at 0, 10, 20, 30, 45 and 60 min after the start of the experiment. A numerical value from 0 to 3 was assigned to each grade for the cataract index (CI) calculation and data analysis. The same procedure was repeated in all groups 48 h later. The reversion process of lens opacity was explored using a hypotonic NaCl solution (100 mOsmol/kg) in another pair of groups, a 500 mOsmol/kg NaCl solution group and natural exposure group. The gross appearance and time course of development and reversion of lens opacity were assessed. RESULTS: Lens opacity primarily developed in a hypertonic NaCl solution-treated and naturally exposed eyes, and the gross anatomical appearance were similar. The speed of lens opacity development and CI changes were osmolarity-dependent, and the higher NaCl concentration solution used, the faster and more severe the formation of opacification. Both hypertonic NaCl-solution-induced lens opacity and natural exposure induced lens opacity could be resolved by hypotonic NaCl solution prior to anesthesia recovery. CONCLUSIONS: This study indicates a crucial effect of NaCl concentration on the development and reversion of lens opacity in the anesthetized mice, and support the osmolarity theory in the reversible lens opacification phenomenon. It is also of practical significance to mouse eye studies that require lens transparency.

Assessment of Full-Eye Response to Osmotic Stress in Mouse Model In Vivo Using Optical Coherence Tomography.

NaCl based solutions were applied as osmotic stress agents to alter the hydration state of the mouse eye. Full-eye responses to these osmotic challenges were monitored in vivo using a custom-built optical coherence tomography (OCT) with an extended imaging range of 12.38 mm. Dynamic changes in the mouse eye were quantified based on the OCT images using several parameters, including the central corneal thickness (CCT), the anterior chamber depth (ACD), the crystalline lens thickness (LT), the cornea-retina distance (CRD), the iris curvature (IC), and the lens scattering intensity (LSI). Apparent but reversible changes in the morphology of almost all the ocular components and the light transparency of the lens are exhibited. Particularly, the ocular dehydration induced by the hypertonic challenges resulted in a closing of the iridocorneal angle and an opacification of the lens. Our results indicated that the ocular hydration is an important physiological process which might be correlated with various ocular disorders, such as dry eye, cataract, and angle-closure glaucoma, and would affect the biometry and imaging of the eye. OCT uniquely enables the comprehensive study of the dynamic full-eye responses to the ocular hydration in vivo.

Identification of a novel mutation in a Chinese family with Nance-Horan syndrome by whole exome sequencing.

OBJECTIVE: Nance-Horan syndrome (NHS) is a rare X-linked disorder characterized by congenital nuclear cataracts, dental anomalies, and craniofacial dysmorphisms. Mental retardation was present in about 30% of the reported cases. The purpose of this study was to investigate the genetic and clinical features of NHS in a Chinese family. METHODS: Whole exome sequencing analysis was performed on DNA from an affected male to scan for candidate mutations on the X-chromosome. Sanger sequencing was used to verify these candidate mutations in the whole family. Clinical and ophthalmological examinations were performed on all members of the family. RESULTS: A combination of exome sequencing and Sanger sequencing revealed a nonsense mutation c.322G>T (E108X) in exon 1 of NHS gene, co-segregating with the disease in the family. The nonsense mutation led to the conversion of glutamic acid to a stop codon (E108X), resulting in truncation of the NHS protein. Multiple sequence alignments showed that codon 108, where the mutation (c.322G>T) occurred, was located within a phylogenetically conserved region. The clinical features in all affected males and female carriers are described in detail. CONCLUSIONS: We report a nonsense mutation c.322G>T (E108X) in a Chinese family with NHS. Our findings broaden the spectrum of NHS mutations and provide molecular insight into future NHS clinical genetic diagnosis.