The classical Hippo pathway leads to the phosphorylation of downstream effector molecules Hippo-Yes-associated protein (Yap) and transcriptional coactivator PDZ-binding motif (Taz) serine sites through a kinase response, thereby promoting cell proliferation, controlling cell polarity, changing cytoskeleton, it plays an important regulatory role in various pathophysiological processes such as epithelial-mesenchymal transition and inhibition of cell contact. Studies have shown that Yap/Taz can affect the progression of vitreoretinal diseases, opening up new prospects for the pathogenesis and clinical treatment of diabetic retinopathy, proliferative vitreoretinopathy, and retinal ischemia-reperfusion injury. Exploring the molecular mechanism of Yap/Taz provides a possible therapeutic target for future research in the treatment of retinal fibrosis diseases such as diabetic retinopathy and proliferative vitreoretinopathy. At the same time, regulating the activity of local Yap/Taz in the retina will also become an effective therapeutic target for damage-repair in retinal ischemia-reperfusion injury. However, Yap inhibitors have potential retinal toxicity and are still in the preclinical development stage. Further research on the mechanism of action and clinical safety of Yap inhibitors will provide new methods for the treatment of retinal diseases.
Microvesicles (MVs) is small membrane vesicles released from different cell types under different conditions. Studies have shown that MVs may mediate vascular inflammation, angiogenesis, and other pathological processes. MVs may play an important role in the pathogenesis of diabetic retinopathy (DR) by mediating endothelial cell injury, thrombosis and neovascularization. The plasma MV level may be an effective parameter to monitor the development of DR. This article will summarize the research progress of the relationship between MVs and DR in recent years.
ObjectiveTo study how CD73 is shed from the retinal pigment epithelium (RPE) surface.MethodsCD73 shedding was induced by treating RPE with lipopolysaccharides (LPS) and TNF-α. After Phospholipase C (PLC) or pan matrix metalloproteinase (MMP) inhibitors were added, surface amount of CD73 was evaluated by flow cytometry (FACS). Then selective inhibitors or their corresponding siRNAs of MMP-2 and MMP-9 were applied to the treatments of RPE; and their effects on induced CD73 shedding were evaluated by FACS. By site directed mutagenesis, mutations were introduced to Lys547-Phe548 coding sites of CD73 cDNA, which was cloned in a pcDNA mammalian expression vector. Both wt-CD73 and mutated-CD73 were over expressed in CD73-/- RPE and their induced shedding was compared.ResultsLPS and TNF-α induced CD73 shedding from RPE was completely blocked by the addition of pan MMP inhibitor but not PLC inhibitor. Selective MPP-9, but not MMP-2, inhibitor or its siRNA blocked CD73 shedding. In CD73-/- RPE induced CD73 shedding was happened to overexpressed wt-CD73 but not Lys547-Phe548 sites mutant CD73.ConclusionMMP-9 is responsible for shedding CD73 from RPE through hydrolyzing its Lys547 -Phe548 sites.
Rituximab (RTX) is a monoclonal antibody directed against the CD20 antigen expressed on B cells. It has been successfully employed in the treatment of non-Hodgkin's lymphoma and varied systemic autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, and granulomatosis with polyangiitis. Recently its efficacy in the treatment of ocular inflammatory diseases (OID), including refractory scleritis, peripheral ulcerative keratitis, uveitis, and ocular cicatricial pemphigoid, has aroused more concerns. The literature suggests that RTX may be useful for controlling the inflammation and decreasing or stopping the use of corticosteroids and other immunosuppressants in OID, which may contribute a new treatment alternative in patients with the recalcitrant and sight-threatening forms of OID. This article reviews the clinical application status of RTX in the treatment of OID.
ObjectiveTo preliminarily investigate the mechanism of MMP-9 blocking CD73 detachment from RPE cells surface and preventing and treating experimental autoimmune pigment membranitis (EAU).MethodsRPE cells isolated from wild-type C57BL/6 and CD73 gene knockout (CD73-/-) mice were cultured in vitro, and treated with lipopolysaccharide and TNF-α to induce CD73 detachment from RPE surface. According to whether MMP-9 inhibitor CTK8G1150 was added at the same time (the final concentration was 5.0 mol/L) or not, RPE cells cultured in the two types of mice were respectively set as MMP-9 inhibitor intervention group and non-intervention control group. The cells in each group were treated with the intervention of a solvent, 1 μmol/L adenosine monophosphate (AMP), 1 μmol/L AMP, and 3 μmol/L 5' -α,β-methylene adenosine diphosphate (APCP) (AMP+APCP). The stimulating effect of RPE cells in different groups on CD4+ T cell proliferation was detected by tritiated thymidine incorporation. Adoptive immune induced EAU in wild-type B6 mice and CD73-/- mice, respectively. The receptor mice were randomly divided into the MMP-9 inhibitor intervention group and the non-intervention control group, and CTK8G1150 or the solvent were injected into the subretinal cavity 4, 7 and 10 days after adoptive immunity. CD73 mRNA and protein expression in RPE cells of recipient mice were detected by real-time quantitative PCR (RT-PCR) and Western blot. One-way ANOVA was used to analyze all experimental data.ResultsWhen the stimulation mode was AMP, the proliferation of CD4+ T cells in the C57BL/6 MMP-9 inhibitor intervention group decreased significantly compared with the non-intervention group (F=13.28, P＜0.01). When the stimulation mode was solvent and AMP+APCP, there was no statistically significant difference in the proliferation capacity of CD4+ T cells between the two groups (F=7.78, 6.58; P＞0.05). There was no statistically significant difference in the proliferation capacity of CD4+ T cells between the CD73-/- MMP-9 inhibitor intervention group and the non-intervention group (F=5.24, 6.12, 7.04; P＞0.05). RT-PCR results showed that there was no statistically significant difference in the relative expression of CD73 mRNA in RPE cells between the MMP-9 inhibitor group and the non-intervention control group (F=6.54, P>0.05). Western blot results showed that the expression of CD73 protein in RPE cells in the MMP-9 inhibitor group of B6 receptor mice was significantly increased compared with the control group (F=15.24, P＜0.01).ConclusionMMP-9 inhibitor blocks CD73 detachment from RPE cells surface and has a protective effect on EAU.
ObjectiveTo observe and analyze the superficial retinal blood flow density and its related influencing factors in the macular area of adolescents with different types of non-pathological high myopia (HM). MethodsA retrospective clinical study. From March to August 2022, 117 eyes of 117 adolescents who were admitted to Liaocheng Aier Eye Hospital due to myopia were included in the study. According to equivalent spherical degree (SE) and corneal curvature, subjects were divided into mild myopia or emmetropia group (control group), HM group, occult HM (OHM) group, and super HM (SHM) group, with 30 eyes, 28 eyes, 35 eyes, and 24 eyes, respectively. All subjects underwent medical optometry, intraocular pressure, optical coherence tomography (OCT), OCT angiography (OCTA), axial length (AL) and corneal curvature measurements. The diopter was SE. OCTA instrument was used to scan the macular region in the range of 6 mm×6 mm, and the software automatically divided it into three concentric circles centered on the fovea of the macular, namely, the central area with a diameter of 1 mm, the inner ring area with a diameter of 1-3 mm, and the outer ring area with a diameter of 3-6 mm. The superficial retinal vascular density (SRVD), vascular perfusion density (SBPD), the area, perimeter (PERIM), avascular index (AI) of foveal avascular area (FAZ) and retinal thickness were measured in the macular region as a whole and in different regions. One-way analysis of variance was used to compare the data among groups, and the least significant difference t-test was used to compare the data among groups. The correlation of AL, corneal curvature and intraocular pressure with SRVD and SBPD in macula was analyzed by Pearson correlation analysis. ResultsThere were significant differences in SRVD and SBPD in the central, inner and outer regions of macula in control group, HM group, OHM group and SHM group (P＜0.05). There were statistically significant differences in the thickness of the retina above, below and on the temporal side of the central and outer ring regions (P＜0.05). However, no statistically significant difference was in the thickness of the retina on the nasal side (P＞0.05). There was no significant difference in PERIM (P＞0.05). There were significant differences in FAZ area and AI (P＜0.05). Correlation analysis showed that AL was negatively correlated with SRVD and SBPD in macular whole and central, inner and outer ring regions (P＜0.05). Corneal curvature and SE were positively correlated with the SRVD and SBPD of macular whole, central area and outer ring area (P＜0.05). AL was negatively correlated with retinal thickness in the outer ring region (P＜0.05). SE was positively correlated with the thickness of the retina above, below and temporally in the outer ring region (P＜0.05). AL was negatively correlated with FAZ area and AI (P＜0.05). SE was positively correlated with FAZ area and PERIM (P＜0.05). Retinal thickness was positively correlated with SRVD and SBPD (P＜0.05). ConclusionsThe SRVD and SBPD of different types of HM in adolescents decreases to different degrees. The thickness of the retina in the central region is thicker, and the retina in the outer ring region is thinner. With the decrease of SRVD, the retinal thickness gradually is thinner.
ObjectiveTo investigate the changes of choroid thickness in adolescents with different types of non-pathological high myopia (HM). MethodsA retrospective observational study. From January 2021 to April 2022, 179 eyes of 101 adolescents with myopia in Liaocheng Aier Eye Hospital were collected and analyzed. According to the spherical equivalent (SE) and corneal curvature, subjects were divided into mild myopia or emmetical eye group (control group), HM group, occult HM group (OHM group) and super HM group (SHM group). There were 52 eyes in 30 cases, 47 eyes in 26 cases, 42 eyes in 24 cases and 38 eyes in 21 cases, respectively. Medical optometry, intraocular pressure, optical coherence tomography (OCT), axial length (AL) and corneal curvature were measured. The macular foveal choroidal thickness was analyzed by using spectral-domain OCT. The diopter was expressed in SE. The thickness of choroid in the fovea of macular region was measured by enhanced deep imaging with frequency domain OCT. The thickness of choroid was measured in 9 regions within 1 mm, 3 mm from the fovea, including the upper, lower, nasal and temporal regions. Generalized estimating equation was used to compare the data among groups, and the least significant difference t-test was used to compare the data among groups. The correlation between AL, corneal curvature, intraocular pressure and choroidal thickness was analyzed by Pearson correlation. ResultsThe choroidal thickness in the foveal macula and the areas 1 mm and 3 mm away from the fovea were compared among the control group, HM group, OHM group and SHM group, the difference were significant (χ2=76.646, 36.715, 27.660, 35.301, 24.346, 38.093, 36.275, 33.584, 36.050; P＜0.05). Compared with the control group, the choroidal thickness of the fovea and the choroidal thickness in each area within 1 and 3 mm from the fovea in the HM group, the OHM group and the SHM group were significantly thinner than those in the control group, and the difference was statistically significant (P＜0.05). There were statistically significant differences in choroidal thickness in each region between the group and the SHM group, and between the OHM group and the SHM group (P＜0.05). The results of correlation analysis showed that AL was negatively correlated with choroidal thickness in various regions (P＜0.05); SE was positively correlated with choroidal thickness in various regions (P＜0.05); corneal curvature and intraocular pressure had no significant correlation with choroidal thickness in various regions (P＞0.05). ConclusionsThe choroidal thickness of SHM is significantly lower than that of OHM and HM; OHM patients have lower SE. However, the choroidal thickness is significantly thinner. AL and SE are the influencing factors of choroidal thickness.
ObjectiveTo investigate the effects of migration and expression from chemokine receptor 4 (chemokine receptor-4, CXCR4) of rat bone marrow mesenchymal stem cells (BMSCs) which were pretreated by atorvastatin (ATV) in vitro.MethodsIsolated, cultivated, identified the BMSCs, pretreated P4-P6 of BMSCs with different concentrations of ATV for 12 hours. The experimental group was divided into control group, 0.1 nM/L (group 0.1 nM), 1 nM/L (1 nM group), 10 nM/L (10 nM group), 100 nM/L (100 nM group), 1 000 nM/L (1 000 nM group). The mRNA and protein of CXCR4 were determined by real time-polymerase chain reaction and Western blot. Immunofluoreseence assay were used to detect the expression levels of CXCR4. The migration ability of BMSCs were measured by transwell chamber.ResultsImmunofluoreseence assay showed the protein level of CXCR4 of group 1 nM and 10 nM were significantly higher than the other group. RT-PCR and Western blot showed the protein and mRNA level of CXCR4 in 10 nM was higher than that in group 1 nM. The migration ability of group 10 nM was higher than 1 nM and control group.ConclusionsATV can be dose-dependent promote expression levels of CXCR4 of BMSCs cultivated in vitro.
ObjectiveTo observe the changes of macular microvessels in patients with retinal vein occlusion (RVO) and macular edema (ME) after intravitreal injection of aflibercept (IVA), and analyze its correlation with best corrected visual acuity (BCVA).MethodsA retrospective case study. Thirty patients (30 eyes) with monocular RVO with ME (RVO-ME) who were diagnosed in the clinical examination of Tianjin Eye Hospital from April 2019 to February 2020 were included in the study. Among them, there were 12 males (12 eyes) and 18 females(18 eyes); the average age was 54.30±13.17 years. The average course of disease was 3.43±1.97 months. Both eyes were examined by BCVA and optical coherence tomography (OCTA). The on-demand injection was adopted after the first injection in IVA treatment regimen. The macular area 6 mm×6 mm in both eyes was scanned with an OCTA instrument, and the area of the foveal avascular area (FAZ), FAZ circumference (PERIM), and out-of-roundness were measured at baseline and 1, 3, and 6 months after treatment. Index (AI), blood flow density within 300 μm width of FAZ (FD-300), foveal retinal thickness (CMT), superficial retinal capillary plexus (SCP), deep retinal capillary plexus (DCP) blood flow density. The paired t test was used to compare the quantitative parameters of the affected eye and the contralateral healthy eye at baseline; the changes of the quantitative parameters at baseline and 1, 3, and 6 months after treatment were analyzed by repeated measures analysis of variance. Pearson correlation analysis was used to analyze the correlation between BCVA, retinal perfusion, and macular blood supply parameters at 6 months after IVA treatment.ResultsAt baseline, compared with the contralateral healthy eye, the FAZ area (t=−4.091), PERIM (t=−5.098) and AI (t=−9.093) of the RVO-ME eye were enlarged, and FD-300 (t=7.237) and overall SCP and DCP blood flow density (t=8.735, 9.897) decreased, the difference was statistically significant (P＜0.001). Six months after treatment, the BCVA of RVO-ME eyes was significantly increased, CMT decreased, FAZ area expanded, and AI decreased (t=8.566, 16.739, −6.469, 9.719; P＜0.001), the difference was statistically significant. There was no significant change in the blood flow density of FD-300 and overall SCP and DCP, and the difference was not statistically significant (t=1.017, 1.197, 0.987; P＞0.05). Compared with baseline, the FAZ area of RVO-ME eyes gradually expanded at 3 and 6 months after treatment, and the difference was statistically significant (F=21.979, P＜0.001). Correlation analysis results showed that BCVA at 6 months after treatment was positively correlated with the overall SCP and DCP blood flow density at baseline and 6 months after treatment (r=−0.538, −0.484, −0.879, −0.854; P＜0.05). There was a negative correlation with the area of FAZ 6 months after treatment (r=0.544, P=0.001). The number of ME recurrences was negatively correlated with BCVA and overall SCP and DCP blood flow density 6 months after treatment (r=0.604, −0.462, −0.528; P＜0.05), it was positively correlated with FAZ area (r=0.379, P=0.043).ConclusionWithin 6 months of IVA treatment in RVO-ME eyes, ME is significantly reduced and visual acuity is improved; SCP blood flow density decreases, and FAZ area expands.
ObjectiveTo evaluate the changes in foveal avascular zone (FAZ) size and foveal microstructures in patients with rhegmatogenous retinal detachment (RRD) and to analyze the correlation between FAZ size, foveal microstructure changes and visual outcomes.MethodsA retrospective study. From January 2017 to January 2019, 79 RRD patients (79 eyes) diagnosed in Department of Ophthalmology in Baoding First Central Hospital were included in the study. The patients were divided into macula-off group (51 eyes), macula-on group (28 eyes). The unaffected fellow eyes were used as a control group. The retinas were reattachment in all eyes after first standard 23G PPV combined with silicone oil filling. Before and 3 months after surgery, all patients underwent a comprehensive ophthalmologic examination, including the measurement of BCVA and OCT angiography (OCTA) examination. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. The FAZ areas were evaluated with OCTA in the superficial capillary plexus layers by using 3 mm×3 mm images of the macular. The following postoperative foveal microstructures were evaluated:epiretinal membranes, macular edema, subretinal fluids, integrity of ellipsoidal zone and external limiting membrane. Pearson correlation analysis was used to analyze the correlation between FAZ area and BCVA. The correlation between macular microstructures and BCVA was analyzed by Spearman correlation analysis.ResultsAt 3 months after the removal of silicone oil, the FAZ area in the macula-off group, macula-on group and the control group were 0.447±0.041, 0.409±0.034, 0.335±0.027 mm2, respectively. Compared with the control group, the FAZ area in the macula-off group and macula-on group were enlarged, the difference was statistically significant (F=184.148, P＜0.05). The logMAR BCVA in the macula-off group and macula-on group were 0.800±0.180 and 0.386±0.160, respectively. There was significant difference in ellipsoidal zone rupture between the two groups (χ2=7.577, P=0.006). The results of correlation analysis showed that the area of FAZ and the incidence of ellipsoidal zone fracture after surgery were positively correlated with BCVA after surgery (r=0.329, 0.290; P=0.003, 0.010).ConclusionsThe FAZ area of RRD is enlarged after surgery. FAZ area and ellipsoid zone fracture are positively correlated with BCVA after surgery.