Cotton RSG2 Mediates Plant Resistance against Verticillium dahliae by miR482b Regulation.

Cotton Verticillium wilt, mainly caused by Verticillium dahliae, has a serious impact on the yield and quality of cotton fiber. Many microRNAs (miRNAs) have been identified to participate in plant resistance to V. dahliae infection, but the exploration of miRNA's function mechanism in plant defense is needed. Here, we demonstrate that the ghr-miR482b-GhRSG2 module mediates cotton plant resistance to V. dahliae infection. Based on the mRNA degradation data and GUS fusion experiments, ghr-miR482b directedly bonds to GhRSG2 mRNA to lead to its degradation. The knockdown and overexpression of ghr-miR482b through virus-induced gene silencing strategies enhanced (decreased by 0.39-fold in disease index compared with the control) and weakened (increased by 0.46-fold) the plant resistance to V. dahliae, respectively. In addition, silencing GhRSG2 significantly increased (increased by 0.93-fold in disease index) the plant sensitivity to V. dahliae compared with the control plants treated with empty vector. The expression levels of two SA-related disease genes, GhPR1 and GhPR2, significantly decreased in GhRSG2-silenced plants by 0.71 and 0.67 times, respectively, and in ghr-miR482b-overexpressed (OX) plants by 0.59 and 0.75 times, respectively, compared with the control, whereas the expression levels of GhPR1 and GhPR2 were significantly increased by 1.21 and 2.59 times, respectively, in ghr-miR482b knockdown (KD) plants. In sum, the ghr-miR482b-GhRSG2 module participates in the regulation of plant defense against V. dahliae by inducing the expression of PR1 and PR2 genes.

The cotton miR530-SAP6 module activated by systemic acquired resistance mediates plant defense against Verticillium dahliae.

Many cotton miRNAs in root responding to Verticillium dahliae infection have been identified. Conversely, the miRNAs in leaf distantly responding to this fungal infection from roots via systemic acquired resistance (SAR) remain to be explored. Here, we constructed two groups of leaf sRNA libraries in cotton treated with V. dahliae via root-dipped method at 7- and 10-day post inoculation. Analysis of high-throughput sRNA sequencing identified 75 known and 379 novel miRNAs, of which 41 miRNAs significantly differentially expressed in fungal treatment plant leaves compared to the mock treatment at two time points. Then we characterized the cotton miR530-SAP6 module as a representative in the distant response to V. dahliae infection in roots. Based on degradome data and a luciferase (LUC) fusion reporter analysis, ghr-miR530 directedly cleaved GhSAP6 mRNA during the post-transcriptional process. Silencing of ghr-miR530 increased plant defense to this fungus, while its overexpression attenuated plant resistance. In link with ghr-miR530 function, the knockdown of GhSAP6 also decreased the plant resistance, resulting from down-regulation of SA-relative gene expression including GhNPR1 and GhPR1. In all, these results demonstrated that there are numerous miRNAs in leaf distantly responding to V. dahliae infection in roots mediate plant immunity.

Circ-ADAM9 Promotes High Glucose-Induced Retinal Pigment Epithelial Cell Injury in DR via Regulating miR-338-3p/CARM1 Axis.

BACKGROUND: Circular RNAs (circRNAs) have been reported to be involved in the regulation of retinal pigment epithelial (RPE) cell injury and are closely related to the development of diabetic retinopathy (DR). More research is needed to confirm the role and mechanism of circ-ADAM9 in DR progression. METHODS: High glucose (HG)-induced RPE cells (ARPE-19) were used to mimic the hyperglycemia condition. The expression of circ-ADAM9, microRNA (miR)-338-3p, and coactivator-associated arginine methyltransferase 1 (CARM1) was measured using quantitative real-time PCR. Cell proliferation and apoptosis were determined using MTT assay, EdU assay, and flow cytometry. The protein expression of apoptosis markers and CARM1 was examined by the western blot analysis. Also, MDA level and SOD activity were determined to assess cell oxidative stress. In addition, the interaction between miR-338-3p and circ-ADAM9 or CARM1 was confirmed by dual-luciferase reporter assay and RIP assay. RESULTS: The expression of circ-ADAM9 was upregulated in DR patients and HG-induced ARPE-19 cells. Silenced circ-ADAM9 could promote proliferation and inhibit inflammation, apoptosis, and oxidative stress in HG-induced ARPE9 cells. In terms of mechanism, circ-ADAM9 could sponge miR-338-3p to upregulate CARM1. The inhibitory effect of circ-ADAM9 knockdown on HG-induced ARPE9 cell injury could be reversed by an miR-338-3p inhibitor. As a target of miR-338-3p, CARM1 knockdown could alleviate HG-induced ARPE9 cells' injury, and its overexpression also could reverse the negatively regulation of miR-338-3p on HG-induced ARPE9 cell injury. CONCLUSION: Circ-ADAM9 contributed to HG-induced ARPE9 cell injury by regulating miR-338-3p/CARM1 axis, which provided effective targets for DR treatment.

Anterior segment OCT application in quantifying posterior capsule opacification severity with varied intraocular lens designs.

AIM: To evaluate the application of anterior segment-optical coherence tomography (AS-OCT) in posterior capsule opacification (PCO) severity assessment and analyse the relationship between PCO severity and intraocular lens (IOL) characters. METHODS: PCO patients were prospectively recruited. Cross-sectional images of the anterior segment at horizontal and vertical meridians were acquired with AS-OCT. The area of the IOL-PC (posterior capsular) space and PCO severity (area, thickness, and density at 3 mm and 5 mm IOL optic regions) were measured. The relationship between PCO severity and visual acuity, comparisons of PCO severity and IOL-PC space using varied IOL designs were analysed. RESULTS: One hundred PCO eyes were enrolled. IOL-PC space, PCO thickness and area were positively correlated with axial length. In addition, PCO area and thickness were positively correlated with visual acuity when it was ≤0.52 logMAR. The cut-off level of visual acuity should be 0.52 logMAR. With varied IOL designs, 3-piece C haptic IOL showed a smaller PCO area and thickness than the 1-piece 3 haptic IOL and 1-piece 4 haptic IOL. PCO area and thickness values for an IOL with a diameter ≤11.0 mm was greater than for an IOL with a diameter of 12.5 mm, and the differences were statistically significant. PCO area and thickness increased when IOL haptic angulation increased (from 0 to 12 degrees). CONCLUSION: In PCO eyes, cut-off level of visual acuity is 0.52 logMAR. With more severe PCO, visual acuity maybe not enough to describe the visual function impairment. PCO severity and IOL-PC space are significantly correlated with axial length and IOL design and material.

Posterior capsular opacification comparison between morphology and objective visual function.

BACKGROUND: To compare the influence of posterior capsule opacification (PCO) morphology and severity on intraocular stray light and visual function with different levels of contrast. METHODS: Forty-five patients diagnosed with PCO were included in this prospective consecutive case series. The Optical Quality Analysis System II (OQAS II) was adopted to assess the objective visual function including objective scatter index (OSI) and optical quality analysis system values (OVs) with 100, 20, and 9% contrast. RTVue-100 OCT was used to evaluate the PCO morphology and severity. Comparisons among visual function, morphology, and severity between pear type and fibrosis PCO were performed. The correlations among the PCO morphology, severity, OSI, and OVs were also determined. RESULTS: There was a significant correlation between increased OSI and decreased visual acuity in PCO patients before laser capsulotomy. The changes of OSI were also correlated with the PCO area for the 3 mm IOL optic region (r = 0.43, p = 0.02). The OSI was significantly higher in pear type PCO when compared with fibrosis PCO (Z = - 4.06, p ≤ 0.001). In addition, the increased OSI in pear type PCO was significantly correlated with the 100% OVs and the 20% OVs but not with the 9% OVs. In fibrosis PCO, OSI was only correlated with the 100% OVs and the 20% OVs pre-YAG. CONCLUSIONS: OSI and OVs could objectively indicate the visual function impairment in PCO patients. Effects of PCO on light scattering and on objective visual function might be explained by the variations of morphology and severity.

Application of 25 MHz B-Scan Ultrasonography to Determine the Integrity of the Posterior Capsule in Posterior Polar Cataract.

PURPOSE: To report the application of 25 MHz B-scan ultrasonography (MHzB) to determine the integrity of the posterior capsule (PC) in posterior polar cataract (PPC). METHODS: Patients with whom PPC was clinically diagnosed using slit lamp microscopy who underwent 25 MHzB before phacoemulsification were retrospectively reviewed. The status of the PC was determined by 25 MHzB before phacoemulsification and confirmed during cataract surgery. RESULTS: In total, 21 eyes in 14 clinically diagnosed PPC patients were enrolled in this study. Out of 25 MHzB images, 19 PCs were found to be intact, while 2 showed dehiscence before cataract surgery. During phacoemulsification, 17 PCs were observed to be intact, while 4 PCs showed posterior capsule rupture (PCR). These 4 PCR cases included the above 2 eyes, in which preexisting dehiscence was detected by 25 MHzB. The other 2 PCR cases showed high reflectivity between high echoes in posterior opacities and the PC, indicating synechia between the PPC and PC. CONCLUSION: This is the first report to show that 25 MHzB can be used to clearly visualize the status of the PC in PPC. These results, in turn, could be used to select the appropriate treatment and to thereby avoid further complications during PPC surgery.

Application of Spectral Domain Optical Coherence Tomography to Objectively Evaluate Posterior Capsular Opacity In Vivo.

OBJECTIVES: To objectively evaluate posterior capsular opacification (PCO) with RTVue-100 spectral domain-OCT and assess the agreement with the Pentacam system. METHODS: Sixty-seven eyes diagnosed with PCO were included. RTVue-100 SD-OCT was used to scan the IOL outline and PCO at horizontal and vertical meridians. PCO was also imaged with a Pentacam and slit-lamp photography system. With RTVue-100 SD-OCT, the PCO area, thickness, density, and objective scores were recorded and used to evaluate the severity of PCO at 3 mm and 5 mm diameter ranges of the IOL optic region. We assessed the correlation of visual acuity, PCO characteristics, and PCO scores. PCO scores acquired from RTVue-100 SD-OCT images were also compared with those from the Pentacam. Differences between pear-type and fibrosis-type PCOs were also compared using RTVue-100 SD-OCT cross-sectional images. RESULTS: The cross-sectional images of PCO acquired with RTVue-100 SD-OCT corresponded well to Pentacam and slit-lamp retroillumination images. IOL-posterior capsular space, area, thickness, and density of the proliferated and accumulated LECs could be clearly visualized and quantified with RTVue-100 SD-OCT. PCO scores were correlated with decreased visual acuity, which was in line with the outcomes using the Pentacam. Differences between the pear-type and fibrosis-type PCO were statistically significant; pear-type PCOs showed a wider and thicker opacification region with lower density compared with fibrosis-type PCOs. CONCLUSION: RTVue-100 SD-OCT could be a powerful tool in PCO objective evaluation and classification. OCT could be used to visualize the morphology and outline of PCO. Thus, it could discriminate and quantify differences between different types of PCO. PCO scores seem to be a useful factor that could reliably reflect PCO severity.