Tetrahedral framework nucleic acids-based delivery of MicroRNA-22 inhibits pathological neovascularization and vaso-obliteration by regulating the Wnt pathway.

The objective of this study was to investigate the effects and molecular mechanisms of tetrahedral framework nucleic acids-microRNA22 (tFNAs-miR22) on inhibiting pathological retinal neovascularization (RNV) and restoring physiological retinal vessels. A novel DNA nanocomplex (tFNAs-miR22) was synthesised by modifying microRNA-22 (miR22) through attachment onto tetrahedral frame nucleic acids (tFNAs), which possess diverse biological functions. Cell proliferation, wound healing, and tube formation were employed for in vitro assays to investigate the angiogenic function of cells. Oxygen-induced retinopathy (OIR) model was utilised to examine the effects of reducing pathological neovascularization (RNV) and inhibiting vascular occlusion in vivo. In vitro, tFNAs-miR22 demonstrated the ability to penetrate endothelial cells and effectively suppress cell proliferation, tube formation, and migration in a hypoxic environment. In vivo, tFNAs-miR22 exhibited promising results in reducing RNV and promoting the restoration of normal retinal blood vessels in OIR model through modulation of the Wnt pathway. This study provided a theoretical basis for the further understanding of RNV, and highlighted the innovative and potential of tFNAs-miR22 as a therapeutic option for ischemic retinal diseases.

Genotype-Phenotype of CRB1-Associated Early-Onset Retinal Dystrophy: Novel Insights on Retinal Architecture and Therapeutic Window for Clinical Trials.

Purpose: The purpose of this study was to investigate the genotypic and phenotypic characteristics of CRB1-associated early onset retinal dystrophy (CRB1-eoRD) and retinal architecture by swept-source optical coherence tomography (SS-OCT). Methods: Eleven probands with CRB1-eoRD were recruited. Clinical information, genetic analysis, and comprehensive ophthalmic examinations including SS-OCT and SS-OCT angiography (SS-OCTA) were conducted. Results: A total of 81.8% (9/11) of CRB1-eoRD presented as Leber congenital amaurosis (LCA). Common clinical manifestations included coin-like yellow-white retinal spots (20/22, 90.9%) and para-arteriolar retinal pigment epithelial retention (12/22, 54.5%). Nineteen different CRB1 variants were detected in our case series, including 12 missense, 3 frameshifts, 3 nonsense, and 1 splicing. Of them, 12 variants had been reported, and 7 were novel. SS-OCT showed thinner central macula (the LCA group, P < 0.0001), thicker total retina (P < 0.0001), thinner outer retina (P < 0.05), and thicker inner retina (P < 0.0001) compared with the healthy control. The inner/outer (I/O) retina thickness ratio of CRB1-eoRD was 3.0, higher than the healthy control of 1.2 and other inherited retinal diseases (IRDs) of 2.2 (P < 0.0001 and P = 0.0027, respectively). SS-OCTA revealed an increased vascular density and perfusion area of the superficial vascular complex and deep vascular complex in CRB1-eoRD. Conclusions: LCA emerges as a frequently occurring phenotype in CRB1-eoRD. The unique features of SS-OCT and SS-OCTA are illustrated, and the novel biomarker, I/O ratio, may facilitate early diagnosis. The insights gained from this study hold significant value in determining the treatment window for potential forthcoming CRB1 gene therapy.

Corneal Higher-Order Aberrations Measurements: Precision of SD-OCT/Placido Topography and Comparison with a Scheimpflug/Placido Topography in Eyes After Small-Incision Lenticule Extraction.

INTRODUCTION: The aim of this study was to evaluate the measurements of corneal higher-order aberrations (HOAs) obtained by a new anterior segment optical coherence tomography (OCT) technique combined with a Placido topographer (the MS-39 device) in eyes with prior small-incision lenticule extraction (SMILE) and compare them to the measurements obtained by a Scheimpflug camera combined with a Placido topographer (the Sirius device). METHODS: A total of 56 eyes (56 patients) were included in this prospective study. Corneal aberrations were analyzed for the anterior, posterior, and total cornea surfaces. Within-subject standard deviation (Sw), test-retest repeatability (TRT), and intraclass correlation coefficient (ICC) were used to assess the intraobserver repeatability and interobserver reproducibility. The differences were evaluated by paired t-test. Bland-Altman plots and 95% limits of agreement (95% LoA) were used to evaluate the agreement. RESULTS: High repeatability was observed for anterior and total corneal parameters, with Sw value < 0.07, TRT ≤ 0.16, and ICCs > 0.893, but not trefoil. For the posterior corneal parameters, ICCs varied from 0.088 to 0.966. Regarding interobserver reproducibility, all Sw values were ≤ 0.04 and TRT ≤ 0.11. ICCs ranged from 0.846 to 0.989, from 0.432 to 0.972, and from 0.798 to 0.985 for the anterior, total, and posterior corneal aberrations parameters, respectively. The mean difference in all aberrations was ≤ 0.05 µm. All parameters showed a narrow 95% LoA. CONCLUSION: The MS-39 device achieved high precision in both anterior and total corneal measurements; the precision of posterior corneal higher-order RMS, astigmatism II, coma, and trefoil was lower. The two technologies used by the MS-39 and Sirius devices can be used interchangeably for measuring corneal HOAs after SMILE.

Repeatability and reproducibility of corneal higher-order aberrations measurements after small incision lenticule extraction using the Scheimpflug-Placido topographer.

BACKGROUND: To evaluate the precision of corneal higher-order aberrations measurements after small incision lenticule extraction (SMILE) using the Sirius Scheimpflug-Placido topographer (CSO, Italy). METHODS: Seventy-five eyes from 75 postoperative subjects were included in this prospective study. Three consecutive corneal aberrometric measurements were obtained with the Scheimpflug-Placido topographer by two experienced operators to assess intra- and inter-observer reproducibility. The within-subject standard deviation (Sw), test-retest repeatability (TRT) and the intraclass correlation coefficient (ICC) were calculated. RESULTS: For intraobserver repeatability of anterior and total corneal aberrations, all ICCs were more than 0.922, except for trefoil (0.722 to 0.768). The ICCs of total root mean square (RMS), coma Z (3, ± 1), and spherical aberration Z (4, 0) were over 0.810 while higher-order RMS, trefoil Z (3, ± 3), and astigmatism II Z (4, ± 2) were below 0.634 for posterior corneal surface aberrations. All Sw values for all types of aberrations were equal to or below 0.07 µm. Regarding interobserver reproducibility, all TRT values were no more than 0.12 µm, 0.05 µm, and 0.11 µm for anterior, posterior, and total corneal aberrations, respectively. The ICC values ranged from 0.875 to 0.989, from 0.686 to 0.976 and over 0.834 for anterior, posterior, and total corneal aberrations, respectively. CONCLUSIONS: The repeatability of measurements of anterior and total corneal aberrations with the Sirius system in corneas after SMILE surgery was high, except for trefoil. There was some variability in posterior corneal aberrometric measurements. High reproducibility of corneal aberrometric measurements was observed between measurements of both examiners, except for trefoil, with poor to moderate reproducibility.

Comprehensive comparisons of ocular biometry: A network-based big data analysis.

PURPOSE: To systematically compare and rank ocular measurements with optical and ultrasound biometers based on big data. METHODS: PubMed, Embase, the Cochrane Library and the US trial registry ( www. CLINICALTRIAL: gov ) were used to systematically search trials published up to October 22nd, 2020. We included comparative studies reporting the following parameters measured by at least two devices: axial length (AL), flattest meridian keratometry (Kf), steepest meridian keratometry (Ks), mean keratometry (Km), astigmatism (AST), astigmatism vectors J0 and J45, anterior chamber depth (ACD), aqueous depth (AQD), central corneal thickness (CCT), corneal diameter (CD) and lens thickness (LT). A network-based big data analysis was conducted using STATA version 13.1. RESULTS: Across 129 studies involving 17,181 eyes, 12 optical biometers and two ultrasound biometers (with both contact and immersion techniques) were identified. A network meta-analysis for AL and ACD measurements found that statistically significant differences existed when contact ultrasound biometry was compared with the optical biometers. There were no statistically significant differences among the four swept-source optical coherence tomography (SS-OCT) based devices (IOLMaster 700, OA-2000, Argos and ANTERION). As for Ks, Km and CD, statistically significant differences were found when the Pentacam AXL was compared with the IOLMaster and IOLMaster 500. There were statistically significant differences for CCT when the OA-2000 was compared to Pentacam AXL, IOLMaster 700, Lenstar, AL-Scan and Galilei G6. CONCLUSION: For AL and ACD, contact ultrasound biometry obtains the lower values compared with optical biometers. The Pentacam AXL achieves the lowest values for keratometry and CD. The smallest value for CCT measurement is found with the OA-2000.

Sirius Scheimpflug-Placido versus ultrasound pachymetry for central corneal thickness: meta-analysis.

BACKGROUND: To compare the difference in central corneal thickness (CCT) measurements in normal eyes between a rotating Scheimpflug camera combined with a Placido-disk corneal topographer (Sirius, CSO, Italy) and ultrasound pachymetry (USP). METHODS: A systematic literature search was conducted for relevant studies published on PubMed, Medline, EMBASE, and the Cochrane Library and ClinicalTrials.gov from inception to August 1st, 2019. Primary outcome measures were CCT measurements between Sirius and USP. A random effects model was used to pool CCT measurements. RESULTS: A total of twelve studies involving 862 eyes were included in this meta-analysis. The meta-analysis found CCT measurements between Sirius and USP to be statistically significantly different (P < 0.0001). The mean difference between Sirius and USP was -11.26 µm with a 95% confidence interval (CI) (-16.92 µm, -5.60 µm). The heterogeneity was I2 = 60% (P = 0.004). CONCLUSION: CCT measurements with the Sirius Scheimpflug-Placido topographer were statistically significantly lower than USP. However, it may be argued that the mean difference of 11.26 µm is not a clinically significant difference.

Repeatability and agreement of corneal thickness measurements by three methods of pachymetry in small incision lenticule extraction eyes.

Background: The accurate evaluation of corneal thickness (CT) post small incision lenticule extraction (SMILE) is clinically relevant to reduce the risk of complications. We aimed to analyze repeatability and agreement of central corneal thickness (CCT), thinnest corneal thickness (TCT), and mid-peripheral corneal thickness (MPCT) measurements using Scheimpflug imaging, anterior segment swept-source optical coherence tomography (AS-SS-OCT, CASIA 1000, Tomey), and ultrasound pachymetry (USP, SP-3000, Tomey) in eyes with previous SMILE. Methods: Ninety-one eyes of 91 patients were included. Within-subject standard deviation (Sw), test-retest repeatability (TRT), intraclass correlation coefficient (ICC), and coefficient of variation (CoV) were used to evaluate repeatability. Agreement was assessed by repeat-measurement analysis of variance and Bland-Altman analysis. Results: The above three instruments revealed that Sw was <5.91 µm, CoV was <1.08%, TRT was <16.38 µm, and ICC was >0.94. The 95% limits of agreement were relatively narrow and Bland-Altman plots were more concentrated at the CCT, at the TCT, and at the CT2mm. However, it was shown to be wide at the CT5mm. Conclusions: The three devices provide good repeatability of CT measurements in patients who undergone SMILE. Agreement between measurements at the CCT, TCT, and CT2mm were high, but measurement agreements among CT5mm revealed poor agreement.

Network Meta-analysis of No-History Methods to Calculate Intraocular Lens Power in Eyes With Previous Myopic Laser Refractive Surgery.

PURPOSE: To systematically compare and rank the predictability of no-history intraocular lens (IOL) power calculation methods after myopic laser refractive surgery. METHODS: PubMed, Embase, the Cochrane Library, and the U.S. trial registry (www.ClinicalTrial.gov) were used to systematically search trials published up to August 2019. Included were case series studies reporting the following outcomes in patients with cataract undergoing phacoemulsification after laser refractive surgery: percentage of eyes with a refractive prediction error (PE) within ±0.50 and ±1.00 diopters (D), mean absolute error (MAE), and median absolute error (MedAE). A network meta-analysis was conducted using the STATA software version 13.1 (STATACorp LLC). RESULTS: Nineteen studies involving 1,098 eyes and 19 formulas were identified. A network meta-analysis for the percentage of eyes with a PE within ±0.50 D found that ray-tracing (Okulix), intraoperative aberrometry (Optiwave Refractive Analysis [ORA]), BESSt, and Seitz/Speicher/Savini (Triple-S) (D-K SRK/T), and Fourier-Domain OCT-Based formulas were more predictive than the Wang/Koch/Maloney, Shammas-PL, modified Rosa, Ferrara, and Equivalent K reading at 4.5 mm using the Double-K Holladay 1 formulas. With regard to ranking, the top four formulas as per the surface under the cumulative ranking curve (SUCRA) values for the percentage of eyes with a PE within ±0.50 D were the Okulix, ORA, BESSt, and Triple-S (D-K SRK/T). With regard to MAE, the ORA showed lower errors when compared to the Shammas-PL formula. In this regard, the top four formulas based on the SUCRA values were the Triple-S, BESSt, ORA, and Fourier-Domain OCT-Based formulas. The SToP (SRK/T), ORA, Fourier-Domain OCT-Based, and BESSt formulas had the lowest MedAE. CONCLUSIONS: Considering all three outcome measures of highest percentages of eyes with a PE within ±0.50 and ±1.00 D, lowest MAE, and lowest MedAE, the top three no-history formulas for IOL power calculation in eyes with previous myopic corneal laser refractive surgery were: ORA, BESSt, and Triple-S (D-K SRK/T). [J Refract Surg. 2020;36(7):481-490.].

The precision and agreement of corneal thickness and keratometry measurements with SS-OCT versus Scheimpflug imaging.

PURPOSE: To assess the repeatability and reproducibility of swept-source optical coherence tomography (SS-OCT) and Scheimpflug system and evaluate the agreement between the two systems in measuring multiple corneal regions in children. METHODS: Pachymetric and keratometric maps for both systems were evaluated. Central, midperipheral and peripheral corneal thickness (CT), keratometry and astigmatism power vectors were recorded. The three outcomes yielded by the same observer were used to assess intraobserver repeatability. The differences in the mean values provided by each observer were used to evaluate interobserver reproducibility. Within-subject standard deviation, test-retest repeatability (TRT) and coefficient of variation (CoV) were used to analyze the intraobserver repeatability and interobserver reproducibility. Paired T-test and Bland-Altman were used to appraise interdevice agreement. RESULTS: Seventy-eight eyes of 78 children were included. The CoV was ≤2.12 and 1.10%, respectively, for repeatability and reproducibility. TRT and CoV were lower for central and paracentral CT measurements than for peripheral measurements. The SS-OCT device generated higher precision when acquiring CT data, whereas Scheimpflug system showed higher reliability when measuring corneal keratometry. Although the CT readings measured using SS-OCT were significantly thinner than Scheimpflug device (P <  0.001), the central and thinnest CT values were still of high agreement. The interdevice agreement of keratometry measurement was high for the central corneal region and moderate for the paracentral and peripheral areas. CONCLUSIONS: The precision of CT measurements by SS-OCT was higher, while the reliability of keratometry measurements by the Scheimpflug system was higher in children. Apart from the measured values in the central corneal region, the thickness and keratometry readings should not be considered interchangeable between the two systems.

Extent of lymph node dissection improves overall survival in pT3N0 non-metastatic renal cell carcinoma patients treated with radical nephrectomy: a propensity score-based analysis.

BACKGROUND: To assess the impact of lymph node dissection (LND) extent on overall survival (OS) and cancer-specific survival (CSS) in patients with pN0M0 renal cell carcinoma (RCC) treated with radical nephrectomy (RN). MATERIALS AND METHODS: Data queried for this study include RCC (2010-2014) from the Surveillance, Epidemiology, and End Results (SEER) program. Kaplan-Meier analyses and multivariate Cox regression models tested the effect of number of removed lymph node (NRN) ≥ 50th percentile on OS and CSS. The associations were evaluated using propensity score (PS) matching adjustment. RESULTS: A total of 5532 pN0M0 RCC patients were enrolled in our study. The median NRN was 2 (IQR 1-6), the 50th percentile defined patients with NRN ≥ 2. Following PS adjustment, there were no significant differences in clinicopathologic features between two groups of patients except for age. Multivariate model analysis showed that patients with NRN < 2 had worse OS than those with NRN ≥ 2 in pT3 group (HR 1.442; P = 0.032) but not in pT1 and pT2 groups (HR 0.859 and 1.393, P = 0.443 and P = 0.267, respectively). However, patients with NRN < 2 had better CCS than those with NRN ≥ 2 in pT1 group (HR 0.368; P = 0.016) but not in pT2 and pT3 groups (HR 1.674 and 1.325, P = 0.216 and P = 0.176, respectively). CONCLUSIONS: More extensive LND (NRN ≥ 2) at RN is associated with better OS in pT3N0M0 RCC patients while it exerts negative influence on CCS in pT1N0M0 group. Hence, more extensive LND has therapeutic value in pT3 individuals but not in pT1 group.

Polydatin exerts anti-tumor effects against renal cell carcinoma cells via induction of caspase-dependent apoptosis and inhibition of the PI3K/Akt pathway.

PURPOSE: Polydatin, a stilbenoid glucoside of a resveratrol derivative, has many biological functions, including antitumor effects. However, the antitumor effects of polydatin in renal cell carcinoma (RCC) have not been investigated. MATERIALS AND METHODS: In the current study, MTT assays, transwell invasion assays and wound healing assays were performed to examine cell proliferation, invasion and migration. An apoptosis nucleosome ELISA was used to measure apoptosis. Caspase activity assays were applied to measure the activities of caspase-3/9. A Western blot assay was used to measure the change in protein levels. RESULTS: Our data demonstrated that polydatin inhibited the proliferation of RCC cells but not normal renal epithelial cells in a time- and dose-dependent manner. Polydatin also triggered apoptosis in a caspase-dependent manner. Moreover, polydatin treatment also led to the downregulation of Bcl-2 and Mcl-1 and to activation of Bax. Ectopic expression of Bcl-2 and Mcl-1 or silencing of Bax could repress the apoptosis that was induced by polydatin. Moreover, incubation with polydatin also suppressed the PI3K/Akt signaling pathway in RCC cells. CONCLUSION: Taken together, our data indicated that polydatin may be applied as a potent agent against RCC.

Methamphetamine exposure triggers apoptosis and autophagy in neuronal cells by activating the C/EBPß-related signaling pathway.

Methamphetamine (Meth) is a widely abused psychoactive drug that primarily damages the nervous system, notably causing dopaminergic neuronal apoptosis. CCAAT-enhancer binding protein (C/EBPß) is a transcription factor and an important regulator of cell apoptosis and autophagy. Insulin-like growth factor binding protein (IGFBP5) is a proapoptotic factor that mediates Meth-induced neuronal apoptosis, and Trib3 (tribbles pseudokinase 3) is an endoplasmic reticulum (ER) stress-inducible gene involved in autophagic cell death through the mammalian target of rapamycin (mTOR) signaling pathway. To test the hypothesis that C/EBPß is involved in Meth-induced IGFBP5-mediated neuronal apoptosis and Trib3-mediated neuronal autophagy, we measured the protein expression of C/EBPß after Meth exposure and evaluated the effects of silencing C/EBPß, IGFBP5, or Trib3 on Meth-induced apoptosis and autophagy in neuronal cells and in the rat striatum after intrastriatal Meth injection. We found that, at relatively high doses, Meth exposure increased C/EBPß protein expression, which was accompanied by increased neuronal apoptosis and autophagy; triggered the IGFBP5-mediated, p53-up-regulated modulator of apoptosis (PUMA)-related mitochondrial apoptotic signaling pathway; and stimulated the Trib3-mediated ER stress signaling pathway through the Akt-mTOR signaling axis. We also found that autophagy is an early response to Meth-induced stress upstream of apoptosis and plays a detrimental role in Meth-induced neuronal cell death. These results suggest that Meth exposure induces C/EBPß expression, which plays an essential role in the neuronal apoptosis and autophagy induced by relatively high doses of Meth; however, relatively low concentrations of Meth did not change the expression of C/EBPß in vitro. Further studies are needed to elucidate the role of C/EBPß in low-dose Meth-induced neurotoxicity.-Xu, X., Huang, E., Luo, B., Cai, D., Zhao, X., Luo, Q., Jin, Y., Chen, L., Wang, Q., Liu, C., Lin, Z., Xie, W.-B., Wang, H. Methamphetamine exposure triggers apoptosis and autophagy in neuronal cells by activating the C/EBPß-related signaling pathway.

Baicalin released from Scutellaria baicalensis induces autotoxicity and promotes soilborn pathogens.

Experiments were conducted to determine whether allelochemicals released by the important medicinal plant Scutellaria baicalensis Georgi help to explain why S. baicalensis performs poorly when continuously cropped. Based on high performance liquid chromatography, the concentration of baicalin (the major compound released by S. baicalensis roots) in the soil where S. baicalensi had been grown for 3 years was 0.97 microg x g(-1). Both the crude extracts from S. baicalensis roots and purified baicalin at 0.97 microg x g(-1) increased the mortality of S. baicalensis seedlings in an autotoxicity test. This concentration stimulated the growth of two soilborne pathogens (Pythium ultimum and Rhizoctonia solani) on agar, and their growth and pathogenic activity in sand. Seedling mortality and damping-off caused by both pathogens were greater in sand where S. canadensis had previously grown than in sand where it had not previously grown. Mortality and damping-off of S. baicalensis seedlings also were significantly higher in soil collected from an S. baicalensis field than in soil collected from a Nicotiana tabacum L. field. The results are consistent with the hypothesis that allelochemicals released by S. baicalensis negatively affect S. baicalensis directly by inducing autotoxicity and indirectly by increasing pathogen activity in the soil.