Identification of aberrantly expressed circular RNAs in hyperlipidemia-induced retinal vascular dysfunction in mice.

Hyperlipidemia-induced retinal vascular dysfunction is a complex pathological process. circRNAs are important regulators of biological processes and disease progression. However, the expression pattern of circRNAs in hyperlipidemia-induced retinal vascular dysfunction remains unclear. Herein, we used a murine model of hyperlipidemia and identified 317 differentially expressed circRNAs between hyperlipidemic retinas and normolipidemic retinas by circRNA microarrays. GO analysis indicated that the host genes of dysregulated circRNAs were targeted to cell differentiation (ontology: biological process), cytoplasm (ontology: cellular component), and protein binding (ontology: molecular function). Pathway analysis revealed that circRNAs-mediated network was mostly enriched in focal adhesion signaling. Notably, circLDB1 was significantly up-regulated in the serum of coronary artery disease patients and aqueous humor of age-related macular degeneration patients. circLDB1 regulated endothelial cell viability, proliferation, and apoptosis in vitro. Thus, circRNAs are the promising targets for the prediction and diagnosis of hyperlipidemia-induced vascular diseases.

Circular Noncoding RNA HIPK3 Mediates Retinal Vascular Dysfunction in Diabetes Mellitus.

BACKGROUND: The vascular complications of diabetes mellitus are the major causes of morbidity and mortality among people with diabetes. Circular RNAs are a class of endogenous noncoding RNAs that regulate gene expression in eukaryotes. In this study, we investigated the role of circular RNA in retinal vascular dysfunction induced by diabetes mellitus. METHODS: Quantitative polymerase chain reactions, Sanger sequencing, and Northern blots were conducted to detect circular HIPK3 (circHIPK3) expression pattern on diabetes mellitus-related stresses. MTT (3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assays, EdU (5-ethynyl-2'-deoxyuridine) incorporation assays, Transwell migration assays, and Matrigel assays were conducted to detect the role of circHIPK3 in retinal endothelial cell function in vitro. Retinal trypsin digestion, vascular permeability assays, and ELISA assays were conducted to detect the role of circHIPK3 in retinal vascular dysfunction in vivo. Bioinformatics analysis, luciferase activity assays, RNA pull-down assays, and in vitro studies were conducted to reveal the mechanism of circHIPK3-mediated retinal vascular dysfunction. RESULTS: circHIPK3 expression was significantly upregulated in diabetic retinas and retinal endothelial cells following stressors related to diabetes mellitus. circHIPK3 silencing or overexpressing circHIPK3 changed retinal endothelial cell viability, proliferation, migration, and tube formation in vitro. circHIPK3 silencing in vivo alleviated retinal vascular dysfunction, as shown by decreased retinal acellular capillaries, vascular leakage, and inflammation. circHIPK3 acted as an endogenous miR-30a-3p sponge to sequester and inhibit miR-30a-3p activity, which led to increased vascular endothelial growth factor-C, FZD4, and WNT2 expression. Ectopic expression of miR-30a-3p mimicked the effect of circHIPK3 silencing on vascular endothelial phenotypes in vivo and in vitro. CONCLUSIONS: The circular RNA circHIPK3 plays a role in diabetic retinopathy by blocking miR-30a function, leading to increased endothelial proliferation and vascular dysfunction. These data suggest that circular RNA is a potential target to control diabetic proliferative retinopathy.

RNCR3 knockdown inhibits diabetes mellitus-induced retinal reactive gliosis.

Retinal reactive gliosis is an important pathological feature of diabetic retinopathy. Identifying the underlying mechanisms causing reactive gliosis will be important for developing new therapeutic strategies for treating diabetic retinopathy. Herein, we show that long noncoding RNA-RNCR3 knockdown significantly inhibits retinal reactive gliosis. RNCR3 knockdown leads to a marked reduction in the release of several cytokines. RNCR3 knockdown alleviates diabetes mellitus-induced retinal neurodegeneration, as shown by less apoptotic retinal cells and ameliorative visual function. RNCR3 knockdown could also decrease Müller glial cell viability and proliferation, and reduce the expression of glial reactivity-related genes including GFAP and vimentin in vitro. Collectively, this study shows that RNCR3 knockdown may be a promising strategy for the prevention of diabetes mellitus-induced retinal neurodegeneration.

Synthesis and cytotoxic evaluation of eremophilane sesquiterpene 07H239-A derivatives.

Nine new derivatives (6-14) of the eremophilane sesquiterpene 07H239-A (5) were designed and semisynthesized with two types of R-groups by amidation. Most of them were active against five human tumor cell lines, and compounds 6-10 were more potent than the natural product 5. In particular, compounds 6 and 9 exhibited the strongest cytotoxic activity against MDA-MB-435 with IC50 values of 0.91 and 0.96 µM, respectively. Preliminary structure-activity relationships (SARs) analysis indicated that the 14-carboxyl in 5 was an ideal target for chemical modification, and the side chain of 5 might play a necessary role in facilitating their cytotoxic potencies.

[Trabeculotomy on primary congenital glaucoma: a retrospective study of 164 cases (257 eyes)].

OBJECTIVE: To study the relationship of long-term effect of trabeculotomy on primary congenital glaucoma and the related risk factors. METHOD: 164 consecutive patients with primary congenital glaucoma (257 eyes), underwent initial surgery of trabeculotomy between 1996 and 2007. Follow-up was conducted for 30.9 (8.6 - 58.3) months, with a follow-up rate of 89.02%. Multivariate analysis by Logistic regression was conducted to analyze the relation of the factors including age of onset, time between onset and operation, preoperative intraocular pressure, clarity of cornea, and corneal diameter to the failure of surgery. Cox proportional hazards regression modeling was used to analyze the factors related to success of surgery. RESULT: Multivariate logistic regression showed that the preoperative intraocular pressure (IOP) and clarity of cornea were independent risk factors for final outcome (OR(IOP) = 1.408, P = 0.047, and OR(CLA) = 1.691, P = 0.019). Cox regression showed that clarity of cornea was the factor related to the surgery success time (OR(CLA) = 1.632, P = 0.008). CONCLUSION: Clarity of cornea reflexes the condition of primary congenital glaucoma more stably than IOP. It is possible to prognosticate the surgical outcome to combine the clarity of cornea with the IOP value before operation.

[Clinical analysis on sixteen cases of Sturge-Weber syndrome-associated glaucoma].

OBJECTIVE: To investigate the clinical features of Sturge-Weber syndrome-associated glaucoma and its surgical treatment. METHODS: A retrospective case series study. The general clinical data and related ocular manifestations in 16 patients (21 eyes) with Sturge-Weber syndrome-associated glaucoma in our hospital from January 2003 to December 2007 were analyzed retrospectively. RESULTS: Age of the patients ranged between 1 month to 31 years old, and the median age was 11 years. Bilateral facial angiomas were present in 8 patients, and two of them had extensive hemangioma. Eleven cases had unilateral glaucoma and 3 of them had bilateral facial angiomas. Five patients had bilateral glaucoma and all of them had bilateral facial angiomas. Open anterior chamber angle was found in all affected eyes by gonioscopy. B-scan ultrasonography was performed in 21 eyes and diffused occupying lesions in the choroid were found in 8 eyes (38.1%). Posterior bowing of the iris, low echo in ciliary body and shallow ciliary body detachment were found by ultrasound biomicroscopy. Anti-glaucoma surgeries including trabeculotomy, trabeculectomy, non-penetrating deep sclerectomy, valve implantation and cyclocryotherapy were performed in 18 eyes. Eighty percent of the eyes which underwent trabeculectomy developed choroidal detachment after operation. Ten patients (11 eyes) were followed-up for 22 months on average. Intraocular pressure was significantly lower than that before the operation (t = 5.3956, P < 0.01). Intraocular pressure in all followed-up eyes was controlled at < or = 21 mm Hg (1 mm Hg = 0.133 kPa). CONCLUSIONS: The clinical features of Sturge-Weber syndrome-associated glaucoma include facial angiomas, open anterior chamber angle and choroidal hemangioma. Anti-glaucoma surgery can reduce the intraocular pressure effectively.

[Observation on the correlation of visual outcome with the position of posterior chamber intraocular lenses after transscleral fixation].

OBJECTIVE: To observe the location of posterior chamber intraocular lenses (PCIOL) after secondary transscleral fixation, to evaluate its impact on the structures of anterior segment and its correlation with visual outcome. METHODS: It was a retrospective case series, included 32 aphakic patients (32 eyes) with secondary transscleral fixation of PCIOL. Postoperatively, the accurate position of the lens and their relationship with adjacent structures were observed by ultrasound biomicroscopy (UBM). The follow-up time period ranged from 6 to 18 months. RESULTS: Best-corrected visual acuity were improved postoperatively in 30 eyes while the other 2 eyes remained unchanged. No tilt or decentration of the lens was demonstrated by the slit lamp microscopy. While using UBM, optic tilt was found in 9 eyes and decentration in 2 eyes. The decentration and tilt of the lens correlated with visual outcome and was statistically significant (X2 = 4.36, P < 0.05). In 12 eyes (37.5%), both haptics were located in the sulcus. In 20 eyes (62.5%), one haptic was located in the sulcus and the other one was in the back of the iris, pars plica (43.8%), pars plana (9.4%) or vitreous cavity (6.3%). The different haptic locations were no correlated with visual outcome ( X2 = 0.26, P > 0.05). CONCLUSIONS: The current suture technique for transscleral fixated PCIOLs can not ensure the proper anatomical outcome. The decentration and tilt of the lens correlate with the postoperative best corrected visual acuity. In our study, we could not find the correlation between the haptic location and visual outcome.

Circular RNA-ZNF609 regulates retinal neurodegeneration by acting as miR-615 sponge.

Glaucoma is a major cause of visual impairment characterized by progressive retinal neurodegeneration. Circular RNAs are a class of endogenous noncoding RNAs that regulate gene expression in eukaryotes. In this study, we investigated the role of cZNF609 in retinal neurodegeneration induced by glaucoma. Methods: qRT-PCR and Sanger sequencing were conducted to detect cZNF609 expression pattern during retinal neurodegeneration. Immunofluorescence staining was conducted to detect the effect of cZNF609 silencing on retinal neurodegeneration in vivo. MTT assay, Ki67 staining, and PI staining were conducted to detect the effect of cZNF609 silencing on retinal glial cells and RGC function in vitro. Bioinformatics analysis, RNA pull-down assays, and in vitro studies were conducted to reveal the mechanism of cZNF609-mediated retinal neurodegeneration. Results: cZNF609 expression was significantly up-regulated during retinal neurodegeneration. cZNF609 silencing reduced retinal reactive gliosis and glial cell activation, and facilitated RGC survival in vivo. cZNF609 silencing directly regulated Müller cell function but indirectly regulated RGC function in vitro. cZNF609 acted as an endogenous miR-615 sponge to sequester and inhibit miR-615 activity, which led to increased METRN. METRN overexpression could partially rescue cZNF609 silencing-mediated inhibitory effects on retinal glial cell proliferation. Conclusion: Intervention of cZNF609 expression is a promising therapeutic strategy for retinal neurodegeneration.

Targeting circular RNA-ZRANB1 for therapeutic intervention in retinal neurodegeneration.

Glaucoma is a neurodegenerative disease characterized by retinal ganglion cell (RGC) loss, optic disc excavation, and progressive visual field loss. Direct or indirect ameliorating retinal neurodegeneration is a promising therapeutic therapy for glaucoma. Circular RNAs (circRNAs) are a class of covalently closed circular RNA transcripts and have emerged as potential regulators in several neurodegenerative diseases. In this study, we show that cZRANB1 expression is significantly upregulated in retinal neurodegeneration induced by glaucoma. cZRANB1 knockdown decreases retinal reactive gliosis, glial cell activation, and facilitates RGC survival in vivo. cZRANB1 knockdown directly regulates Müller cell function and indirectly regulates RGC function in vitro. cZRANB1 acts as miRNA sponge to regulate Müller cell function through cZRANB1/miR-217/RUNX2 network. Intervention of cZRANB1 expression would become an effective strategy for treating retinal neurodegeneration.

Silencing Of Circular RNA-ZNF609 Ameliorates Vascular Endothelial Dysfunction.

Vascular dysfunction is a hallmark of ischemic, cancer, and inflammatory diseases, contributing to disease progression. Circular RNAs (circRNAs) are endogenous non-coding RNAs, which have been reported to be abnormally expressed in many human diseases. In this study, we used retinal vasculature to determine the role of circular RNA in vascular dysfunction. We revealed that cZNF609 was significantly up-regulated upon high glucose and hypoxia stress in vivo and in vitro. cZNF609 silencing decreased retinal vessel loss and suppressed pathological angiogenesis in vivo. cZNF609 silencing increased endothelial cell migration and tube formation, and protected endothelial cell against oxidative stress and hypoxia stress in vitro. By contrast, transgenic overexpression of cZNF609 showed an opposite effects. cZNF609 acted as an endogenous miR-615-5p sponge to sequester and inhibit miR-615-5p activity, which led to increased MEF2A expression. MEF2A overexpression could rescue cZNF609 silencing-mediated effects on endothelial cell migration, tube formation, and apoptosis. Moreover, dysregulated cZNF609 expression was detected in the clinical samples of the patients with diabetes, hypertension, and coronary artery disease. Intervention of cZNF609 expression is promising therapy for vascular dysfunction.

A proper triage for detecting women with high-risk human papillomavirus genotypes other than HPV16/18.

OBJECTIVE: To evaluate the effect of human papillomavirus (HPV) L1 capsid protein detection in cervical exfoliated cells as a proper triage for women with high-risk human papillomavirus (hrHPV) genotypes other than HPV 16/18. STUDY DESIGN: From January 2013 to June 2015, a total of 513 women aged 30-65 years infected with non-16/18 hrHPV were enrolled into the study. Primary HPV testing, HPV 16/18 genotyping and Papanicolaou (Pap) test were performed in all eligible women. HPV L1 capsid protein was detected by immunocytochemistry in cervical exfoliated cells. All hrHPV positive women underwent colposcopy and further biopsy if indicated. Relationships between HPV L1 capsid protein expression and histologic diagnosis, as well as cytology were analyzed. RESULTS: The positive expression rate of HPV L1 capsid protein in CIN2+ group was significantly lower than that in CIN1 or better group (16.3% vs. 66.7%, P=0.000). Compared with the Pap test, HPV L1 detection achieved higher sensitivity (83.7% vs. 51.2%, P=0.000), higher negative predictive value (NPV) (95.3% vs. 88.6%, P=0.002), and significant lower specificity (66.7% vs.76.1%, P=0.002) while identifying CIN2+. Compared with the Pap test, HPV L1 detection achieved higher sensitivity (89.7% vs. 51.7%, P=0.008), higher NPV (99.0% vs. 96.2%, P=0.043), and significant lower specificity (61.2% vs.73.8%, P=0.000) while identifying CIN3+. CONCLUSION: Because of its higher sensitivity and NPV than that of Pap test, HPV L1 capsid protein detection in cervical exfoliated cells reduces the missed identification of high-grade cervical intraepithelial neoplasia (CIN) in women with hrHPV genotypes other than HPV 16/18. HPV L1 capsid protein detection could be a potential triage for women with non-16/18 hrHPV infection.

The Biogeographic South-North Divide of Polygonatum (Asparagaceae Tribe Polygonateae) within Eastern Asia and Its Recent Dispersals in the Northern Hemisphere.

Eastern Asia (EA) is a key region for the diversification of flowering plants in the Northern Hemisphere, but few studies have focused on the biogeographic history within EA in the context of the other northern continents. Polygonatum is an important medicinal genus widely distributed in the Northern Hemisphere with its highest species richness in EA, and it represents an excellent model for studying the evolution of biogeographic patterns in this region. Divergence time estimation was used to examine the biogeographic history of Polygonatum based on nuclear ITS and four plastid sequences (rbcL, matK, psbA-trnH and trnC-petN) from 30 Polygonatum species and 35 outgroup taxa. The ancestral area of Polygonatum and subsequent dispersal routes were inferred using Bayes-Lagrange. Polygonatum was estimated to have originated in southern EA during the middle Miocene (14.34-13.57 Ma) with subsequent south-to-north expansion in the late Miocene. Multiple intercontinental dispersal events were inferred between EA and Europe or North America, and all of them have occurred recently in the late Miocene to Pliocene. The separation of Polygonatum into the south and north lineages and their subsequent diversifications in the late Miocene supports the existence of a biogeographic divide between the northern and southern parts of EA that also coincides with the retreat and redevelopment of the arid zone in EA in the Neogene. Our results demonstrate the complexity of biogeographic history of Polygonatum in the Northern Hemisphere including early vicariance followed by frequent and recent dispersals in the Neogene.

Long Noncoding RNA-GAS5: A Novel Regulator of Hypertension-Induced Vascular Remodeling.

Vascular remodeling is an important pathological feature of hypertension, leading to increased vascular resistance and reduced compliance. Endothelial cell (EC) and vascular smooth muscle cell (VSMC) dysfunction is involved in vascular remodeling. Long noncoding RNAs are potential regulators of EC and VSMC function. Herein, we determined whether long noncoding RNA-growth arrest-specific 5 (GAS5) is involved in hypertension-related vascular remodeling. We revealed that GAS5 knockdown aggravated hypertension-induced microvascular dysfunction as shown by increased retinal neovascularization and capillary leakage. GAS5 regulated the remodeling of arteries, including caudal arteries, carotid arteries, renal arteries, and thoracic arteries. GAS5 was mainly expressed in ECs and VSMCs, and its expression was significantly downregulated in hypertension. GAS5 knockdown affected endothelial activation, endothelial proliferation, VSMC phenotypic conversion, and EC-VSMC communication in vivo and in vitro. Mechanistically, GAS5 regulated EC and VSMC function through ß-catenin signaling. This study identified GAS5 as a critical regulator in hypertension and demonstrated the potential of gene therapy and drug development for treating hypertension.

Diterpenoids from the roots of Croton crassifolius and their anti-angiogenic activity.

Six diterpenoids [crassifolin J, K, L, M, N and O] along with eleven known ones were isolated from the supercritical fluid extract (SFE) of the roots of Croton crassifolius (Euphorbiaceae). Their structures were elucidated using spectroscopic methods (IR, UV, HRESIMS, 1D and 2D NMR). The structure and stereochemistry of crassifolin J was confirmed by single-crystal X-ray diffraction analysis, and the absolute configurations of crassifolin K-M were determined by CD spectra. Twenty-three diterpenoids from this plant were screened for their anti-angiogenic activity using a wild-type zebrafish in vivo model. Four of the known compounds were active, of which penduliflaworosin possessed the best activity relative to the positive control (SU5416). Further study demonstrated that penduliflaworosin could inhibit vessel formation on Tg(fli1a:EGFP)y1-type zebrafish embryos.

[Black diaphragm intraocular lens for congenital aniridia or traumatic iris deficiency].

OBJECTIVE: To evaluate the efficacy of a black diaphragm intraocular lens (IOL) implantation for congenital aniridia or traumatic iris deficiency and to explore the mechanism of postoperative complications. METHODS: The therapeutic effects and postoperative complications of twenty-five cases (27 eyes) of black diaphragm intraocular lens implantation were analyzed. Ultrasound biomicroscope (UBM) and gonioscopy were used to observe the position of the IOL haptics and anterior chamber angle structure in patient who developed secondary glaucoma postoperatively. RESULTS: After 3-18 months of follow-up, photophobia was reduced or disappear. The best corrected visual acuity (BCVA) Postoperation was improved 2 lines in 20 eyes, no change or within 1 line in 5 eyes, decreased 2 lines or more in 2 eyes. The complications included secondary glaucoma (10 eyes, 37.0%), astigmatism (17 eyes, 63.0%), corneal decompensation (2 eyes, 7.4%), hyphema (2 eyes, 7.4%), vitreous hemorrhage (3 eyes, 11.1%), and retinal detachment (1 eye, 3.7%). Of the 20 haptics in 10 patients, 9 were adequately located in the sulcus region, the others were located in chamber angle, ciliary processes or posterior to the ciliary processes. The changes of chamber angle included angle recession, anterior adhesion of iris and angle closure. CONCLUSIONS: The black diaphragm intraocular lens implantation provides an effective method in the treatment of congenital aniridia or traumatic iris deficiency. Chamber angle impair is major cause of postoperative secondary glaucoma. Abnormality of lens haptics position, hemorrhage and persistent inflammation were also play a role in secondary glaucoma.

[The failure causes of non-penetrating trabecular surgery and reoperation].

OBJECTIVE: To study the failure causes of non-penetrating trabecular surgery (NPTS) with SKGeL (a hyaluronic acid biological gel) implant and the surgical method of reoperation. METHODS: Repeated operation with mitomycin (MMC) through the initial surgical site was performed on 13 failure cases (13 eyes) that had undergone NPTS with SKGeL implant. The blockage of filtration tract was removed and the anterior chamber was intact during the surgery. All of these cases were open-angle glaucoma. Before the repeated surgery ultrasound biomicroscopic (UBM) examination was performed on the primary filtering bleb, and the intraocular pressure (IOP) examination was followed after the repeated operation. RESULTS: The mean follow-up period was (14.0 +/- 5.8) months (6 to 24 months). The examination of UBM showed that the filtering bleb disappeared and there was a liquid chamber under the superficial scleral flap in every failure case. The filtration failure due to the scarring at conjunctiva-Tenon's capsule-superficial scleral flap interface in 9 cases, proliferative membrane formation on the trabecular surface in 3 cases, micro-penetration of the trabecula in 1 case. At the end of follow-up, the IOP of 10 cases was lower than 21 mmHg without medication, the mean IOP level was (14.1 +/- 3.7) mm Hg, the IOP of 1 cases was 15 mmHg with Betagen, another 2 cases failed again 6 months after the repeated surgery and underwent the trabeculectomy at last. The complications included hyphema in one case and micro-penetration of the tabecula in one case. CONCLUSIONS: The scarring at conjunctiva-Tenon's capsule-superficial scleral flap interface was the most important cause of NPTS with SKGeL implant failure. A repeated surgery with MMC through the initial surgical site may be a choice for the failure cases.