FGF1ΔHBS ameliorates retinal inflammation via suppressing TSPO signal in a type 2 diabetes mouse model.

Translocator protein (18 kDa) (TSPO) plays an important role in retinal neuroinflammation in the early stage of diabetic retinopathy (DR). Studies have found that a FGF1 variant (FGF1ΔHBS) with reduced proliferative potency exerts excellent anti-inflammatory effects and potential therapeutic value for diabetic complications. In this study, intravitreal injection of FGF1ΔHBS was administrated every week for one month in db/db mice, which are genetically predisposed to develop type 2 diabetes mellitus and early retinopathy. Changes in retinal function and structure in the animal models were detected by electrophysiology (ERG) and optical tomography coherence (OCT). TSPO expression and retinal inflammation were analyzed by immunofluorescence, Western blot and real-time qPCR. In the retina of T2D (db/db) mice, FGF1 was significantly down-regulated while FGFR1 was up-regulated (both p < 0.05). TSPO and retinal inflammatory factors were all up-regulated. TSPO and FGFR1 were mainly co-stained in the inner retina. After FGF1ΔHBS treatment, ERG showed that the total amplitude of dark-adapted b-wave and oscillating potentials (Ops) was significantly improved, and OCT showed that the thickness of the retina around the optical nerve head was significantly preserved in T2D mice (all p < 0.05). The TSPO signal was significantly suppressed by FGF1ΔHBS. The activation of NF-κB p65 and the expression of inflammatory factors such as TNF-α, IL-1ß, IL-6, COX-2, MIP-1α, and iNOS were all significantly down-regulated (all p < 0.05). Collectively, our current data demonstrated that intravitreal FGF1ΔHBS treatment can effectively inhibit retinal inflammation via suppressing TSPO signal and to preserve retinal function and structure in a T2D mouse model.

Universal and hypoimmunogenic pluripotent stem cells for clinical usage.

It is urgent to prepare and store large numbers of clinical trial grade human pluripotent stem (hPS) cells for off-the-shelf use in stem cell therapies. However, stem cell banks, which store off-the-shelf stem cells, need financial support and large amounts of technicians for daily cell maintenance. Therefore, it is valuable to create "universal" or "hypoimmunogenic" hPS cells with genome editing engineering by knocking in or out immune-related genes. Only a small number of universal or hypoimmunogenic hPS cell lines should be needed to store for off-the-shelf usage and reduce the large amounts of instruments, consumables and technicians. In this article, we consider how to create hypoimmunogenic or universal hPS cells as well as the demerits of the technology. ß2-Microglobulin-knockout hPS cells did not harbor human leukocyte antigen (HLA)-expressing class I cells but led to the activation of natural killer cells. To escape the activities of macrophages and natural killer cells, homozygous hPS cells having a single allele of an HLA class I gene, such as HLA-C, were proposed. Major HLA class Ia molecules were knocked out, and CD47, HLA-G and PD-L1 were knocked in hPS cells utilizing CRISPR/Cas9 genome editing. Finally, some researchers are trying to generate universal hPS cells without genome editing. The cells evaded the activation of not only T cells but also macrophages and natural killer cells. These universal hPS cells have high potential for application in cell therapy.

Next-generation materials for RNA-lipid nanoparticles: lyophilization and targeted transfection.

RNA, including mRNA, siRNA and miRNA, is part of a new class of patient treatments that prevent and treat several diseases. As an alternative to DNA therapy using plasmid DNA, RNA functions in the cellular cytosol, avoiding the potential risks of insertion into patient genomes. RNA drugs, including mRNA vaccines, need carrier materials for delivery into the patient's body. Several delivery carriers of mRNA, such as cationic polymers, lipoplexes, lipid-polymer nanoparticles and lipid nanoparticles (LNPs), have been investigated. For clinical applications, one of the most commonly selected types of RNA delivery carrier is LNPs, which are typically formed with (a) ionizable lipids, which bind to RNA; (b) cholesterol for stabilization; (c) phospholipids to form the LNPs; and (d) polyethylene glycol-conjugated lipids to prevent aggregation and provide stealth characteristics. Most RNA-LNP research has been devoted to achieving highly efficient RNA expression in vitro and in vivo. It is also necessary to study the extended storage of RNA-LNPs under mild conditions. One of the most efficient methods to store RNA-LNPs for a long time is to prepare freeze-dried (lyophilized) RNA-LNPs. Future research should include investigating LNP materials for the development of freeze-dried RNA-LNPs using optimal lipid components and compositions with optimal cryoprotectants. Furthermore, the development of sophisticated RNA-LNP materials for targeted transfection into specific tissues, organs or cells will be a future direction in the development RNA therapeutics. We will discuss the prospects for the development of next-generation RNA-LNP materials.

The relationship between the neutrophil-to-lymphocyte ratio and diabetic retinopathy in adults from the United States: results from the National Health and nutrition examination survey.

BACKGROUND: Diabetic retinopathy (DR) is a common complication of diabetes mellitus (DM). Systemic inflammation is intimately associated with DR. The neutrophil-to-lymphocyte ratio (NLR) index is a relatively new indicator of inflammation. METHODS: This cross-sectional study was carried out among adults with DM based on the National Health and Nutrition Examination Survey (NHANES) from 2009 to 2016. NLR was presented as absolute neutrophil counts/ absolute lymphocyte counts. The relationship of NLR levels to DR was analyzed using multivariable logistic regression. RESULTS: There were 2772 eligible subjects extracted from the NHANES. In the multivariate analysis, NLR was related to the risk of DR after adjustment for potential confounders. The association between NLR levels and DR was nonlinear, with an inflection point of 4.778. Compared with the baseline values, NLR was not statistically significant on the right side of the inflection point (1.000, 0.914 to 1.094, 0.9974) but was positively associated with DR on the left side (1.236, 1.132 to 1.349, < 0.0001). CONCLUSIONS: NLR reflects systemic inflammation that may increase the risk of DR. NLR positively correlates with DR when its value is less than 4.778.

The neutrophil percentage-to-albumin ratio is related to the occurrence of diabetic retinopathy.

BACKGROUND: Among patients with diabetic retinopathy (DR), no proof was available to confirm the prognostic significance of the neutrophil percentage-to-albumin ratio (NPAR). We hypothesized that NPAR plays a role in the incidence of DR in diabetic patients. METHODS: We extracted all diabetes mellitus (DM) data from the National Health and Nutrition Examination Survey (NHANES) database between 1999 and 2018, NPAR was expressed as neutrophil percentage/albumin. Multivariable logistic regression and generalized additive model were utilized for the purpose of examining the correction between NPAR levels and DR. Subgroup analysis of the associations between NPAR and DR was carried out to investigate if the impact of the NPAR varied among different subgroups. RESULTS: An aggregate of 5850 eligible participants were included in the present research. The relationship between NPAR levels and DR was positive linear. In the multivariate analysis, following the adjustment for confounders (gender, white blood cell, age, monocyte percent, red cell distribution width, eosinophils percent, bicarbonate, body mass index, iron, glucose, basophils percent, total bilirubin, creatinine, and chloride), higher NPAR was an independent risk factor for DR compared to lower NPAR (OR, 95% CI: 1.18, 1.00-1.39; 1.24, 1.04-1.48). For the purpose of sensitivity analysis, we found a trend of consistency (p for trend: 0.0190). The results of the subgroup analysis revealed that NPAR did not exert any statistically significant interactions with any of the other DR risk variables. CONCLUSIONS: Elevated NPAR is associated with an elevated risk of occurrence of DR in diabetic patients.

Identification and functional annotation of differentially expressed long noncoding RNAs in retinoblastoma.

Retinoblastoma (RB), the most common intraocular malignancy, typically occurs in pediatric patients under the age of 6 years. The present study aimed to explore the long noncoding RNA (lncRNA) expression profile in RB and identify novel lncRNA biomarkers to facilitate the investigation of molecular mechanisms of RB and improve clinical therapy. Raw microarray data for the comparison of gene expression between three RB and three adjacent normal tissue samples were downloaded from Gene Expression Omnibus (dataset no. GSE111168). After identification of differentially expressed lncRNAs (DELs) and differentially expressed mRNAs (DEMs) in RB, functional enrichment analyses and a DEL-DEM weighted correlation network analysis were performed. A total of 3,915 DELs (1,774 upregulated and 2,141 downregulated) and 3,715 DEMs (1,492 upregulated and 2,223 downregulated) were identified in RB. The DEL-targeted DEMs were highly enriched by genes involved in hexose transport, muscle tissue morphogenesis, the stereocilium membrane, endothelin B receptor binding and γ-filamin/ABP-L, α-actinin and telethonin binding protein of the Z-disc binding. Furthermore, associations of the DELs and DEMs with several pathways were determined, including PI3K/AKT, Hippo and cancer signaling, as well as extracellular matrix-receptor interaction pathways. Coexpression network analysis revealed that the top three DELs, lnc-DAZ1-161, lnc-HDAC7-21 and lnc-OR52A1-55, formed coexpression modules with 181, 156 and 210 DEMs, respectively. In addition, the top three DEMs, namely EIF1AY, GSTM1 and NLRP11, formed coexpression modules with 33, 50 and 41 DELs, respectively. Validation using reverse transcription-quantitative PCR indicated that the expression of representative lncRNAs (lnc-DAZ1-161 and lnc-HDAC7-21) in RB cells in vitro was consistent with that in RB tissues in the database, while the expression of lnc-OR52A1-55 was not consistent with the database. These results suggested that the aberrant lncRNA expression profile in RB is related to the differential regulation of numerous physiological and pathological processes. The lncRNA and mRNA profiles in RB identified may provide novel targets for the investigation of its molecular mechanisms and thus lead to improvements in clinical therapy for RB.

Poly(vinyl alcohol-co-itaconic acid) hydrogels grafted with several designed peptides for human pluripotent stem cell culture and differentiation into cardiomyocytes.

We developed poly(vinyl alcohol-co-itaconic acid) (PV) hydrogels grafted with laminin-derived peptides that had different joint segments and several specific designs, including dual chain motifs. PV hydrogels grafted with a peptide derived from laminin-ß4 (PMQKMRGDVFSP) containing a joint segment, dual chain motif and cationic amino acid insertion could attach human pluripotent stem (hPS) cells and promoted high expansion folds in long-term culture (over 10 passages) with low differentiation rates, whereas hPS cells attached poorly on PV hydrogels grafted with laminin-α5 peptides that had joint segments with and without a cationic amino acid or on PV hydrogels grafted with laminin-ß4 peptides containing the joint segment only. The inclusion of a cationic amino acid in the laminin-ß4 peptide was critical for hPS cell attachment on PV hydrogels, which contributed to the zeta potential shifting to higher values (3-4 mV enhancement). The novel peptide segment-grafted PV hydrogels developed in this study supported hPS cell proliferation, which induced better hPS cell expansion than recombinant vitronectin-coated dishes (gold standard of hPS cell culture dishes) in xeno-free culture conditions. After long-term culture on peptide-grafted hydrogels, hPS cells could be induced to differentiate into specific lineages of cells, such as cardiomyocytes, with high efficiency.

Comparative study on optic disc features of premature infants and full-term newborns.

BACKGROUND: To study optic disc features of premature infants and compare to that of term infants to explore the pattern and features of newborn optic disc development and provide the basis for the diagnosis of newborn optic disc disease. METHODS: This was a prospective clinical research. Newborns underwent newborn fundus disease screening from January 1st, 2016 to October 31st, 2016 in the neonatal ward of Ruian City Maternal and Child Health Hospital were selected. RetCam 3 Version6.1.25.0 Wide-Field Digital Pediatric Retinal Imaging System developed by Clarity Medical Systems, Inc was adopted to conduct fundus examination on both eyes, 130 degree wide-angle lens was used to film the images centering optic disc. RESULTS: For both premature infants and full-term newborns, vertical diameter of the optic disc to lateral diameter of the optic disc ratio was > 1, and the shape of the optic disc was a vertical oval. The difference of each optic disc parameter between premature infants and full-term newborns was not statistically significant (P > 0.05). There's a difference of constitution of sclerotic ring type on optic disc between premature infants and full-term newborns. Among which, the proportion of single ring type and double ring type in premature infants was higher than that in full-term newborns (P < 0.05). The proportion of no ring type in full-term newborns was higher than that in premature infants (P < 0.05). The proportion of mixed type had no significant difference (P > 0.05) between premature infants and full-term newborns. CONCLUSIONS: We found that The proportion of mature types (single ring type and double ring type) in full-term newborns was higher than that in premature infants. While there's no statistical difference of the proportion of mixed types between premature infants and full-term newborns. Double ring type was a normal stage of the development of optic disc.

NOVEL THREE TYPES OF NEOVASCULARIZATION ELSEWHERE DETERMINE THE DIFFERENTIAL CLINICAL FEATURES OF PROLIFERATIVE DIABETIC RETINOPATHY.

PURPOSE: To explore the pathological features and clinical significance of three types of neovascularization elsewhere (NVE) in proliferative diabetic retinopathy. METHODS: Neovascularization elsewhere was classified based on the origins and morphologic features using fluorescein angiography and angiographic and structural optical coherence tomography. The topographical distribution, vitreoretinal interface, and responsiveness to panretinal photocoagulation were compared among three types of NVE. RESULTS: One hundred and twenty-seven NVEs were classified into three types. Type 1 NVE was concentrated along or adjacent to vascular arcades; Type 2 was distributed more peripherally than were Types 1 and 3 NVE. The arch bridge-like vitreoretinal interface accounted for 79% of Type 1 NVE. The flat and flat-forward vitreoretinal interface accounted for 95% and 100% in Type 2 and Type 3 NVE, respectively. At 3 months after panretinal photocoagulation, the regression rates for Types 1, 2, and 3 NVE were 82%, 100%, and 80%, respectively. Type 2 NVE showed best regression rate after panretinal photocoagulation (both P < 0.01). CONCLUSION: Three types of NVE determine the distinctly topographical distributions, vitreoretinal interface features, and differential responsiveness to panretinal photocoagulation treatment. This new concept may have important clinical implications in assessing the treatment and prognosis of proliferative diabetic retinopathy.

Culture and differentiation of purified human adipose-derived stem cells by membrane filtration via nylon mesh filters.

Human adipose-derived stem cells (hASCs) cultured for 5 passages were filtered through nylon (NY) mesh filter membranes coated with and without extracellular matrix proteins to obtain the permeation solution. Subsequently, the culture media were filtered via the membranes to obtain the recovery solution. Then, the membranes were cultured in cell culture medium to obtain the migrated cells from the membranes. The hASCs in the permeation solution, through any type of NY mesh filter membrane having 11 and 20 µm pore sizes, had lower osteogenic differentiation ability than conventional hASCs cultured on tissue culture polystyrene (TCP) dishes for passage 5, whereas the hASCs purified by the membrane migration method through NY mesh filter membranes coated with recombinant vitronectin, which have 11 and 20 µm pore sizes, showed a higher proliferation speed as well as higher osteogenic differentiation potential than the conventional hASCs cultured on TCP dishes for passage 5. The membrane filtration and migration methods would be useful for cell sorting for specific cells, such as hASCs with high proliferation and high osteogenic differentiation ability, which do not need antibody binding or genetic modification of the cells for the specific isolation of the cells.

Comparative Study on the Efficacy and Safety of Tumor Resection in Vitrectomy for Retinal Vasoproliferative Tumors.

PURPOSE: To investigate the efficacy and safety of combined vitrectomy with tumor resection in the treatment of retinal vasoproliferative tumors (RVPT). METHODS: Retrospective study. RVPT patients who underwent vitreous surgery at the Eye Hospital of Wenzhou Medical University from January 2011 to July 2017 were included. The main outcomes included treatment type, tumor activity, and best-corrected visual acuity (BCVA). RESULTS: Altogether, 16 patients with 17 eyes were enrolled with follow-up of no less than 6 months. Eight eyes were in the resection treatment group (Group R) and 9 eyes were in the conservative treatment group (Group C). Female (69%) were more common. The mean age was 50 (49.72 ± 12.92) years. Fifteen patients got unilateral onset and only one patient suffered bilaterally. The common symptoms were decreased visual acuity, floaters, and visual distortion. The preoperative BCVA ranged from hand movement to 20/20, with an average of 0.82 ± 0.75 LogMAR. Patients were all not high myopia, with a mean axial length of 23.27 ± 0.27 mm (21.61 mm to 24.67 mm). Of the retinal diseases, the epiretinal membrane was the most common, followed by vitreous hemorrhage, uveitis, subretinal fluid, and so on. Compared with the baseline BCVA, it improved more at postoperative 6 months and the last visit in Group R than in Group C (P=0.006 and P=0.033). The BCVA-improved 0.2 LogMAR or above in 6 months was 2 eyes in Group C and 7 eyes in Group R. All tumors in Group R were completely resected, whereas three in Group C (33.3%) had definite activity (P=0.008). In all samples, tumors were located on the inner side of the retina and had small vessel wall thickening and hyaline degeneration. The degree of astrocyte proliferation varies widely among different tumors. CONCLUSIONS: RVPT was more likely to occur in nonhigh myopia patients. Epiretinal membrane and vitreous hemorrhage were the main causes for vitreous surgery in RVPT patients. Compared with conservative treatment, surgical resection of the tumor is more beneficial to patients on visual acuity recovery and preventing tumor relapse. It is a safe and effective way to treat RVPT.

The efficacy and safety of Retcam in detecting neonatal retinal hemorrhages.

BACKGROUND: To investigate the ability of characterizing neonatal retinal hemorrhage (RH) using RetCam in healthy newborns and the systemic effects during the procedure. METHODS: This prospective study enrolled 68 healthy newborns aged 2 to 4 days old. The RH was imaged and classified according to the location and numbers of hemorrhages. The heart rate (HR), respiration rate (RR), and oxygen saturation (OS) were recorded at 4 time points before (Phase 1, P1), during (P2 and P3) and after the examination (P4). RESULTS: The median exam time was 151 s. RH was present in 15 infants and 23 eyes. All 23 eyes had hemorrhage in Zone II. Grade II and III hemorrhages were present in 5 and 18 eyes, respectively. The HR increased to 168 beats per minute (bpm) in P3 and recovered to 122.5 bpm in P4. The RR increased to 38 bpm in P3 and recovered to 25 bpm in P4. The OS was reduced to 83% in P2 and recovered to 96% in P4. CONCLUSIONS: RH in healthy newborns, mostly present in Zone II with grade II and III, can be characterized in detail by RetCam. Systemic effects during the process are mild and can be revolved spontaneously.

Characteristics of Neovascularization in Early Stages of Proliferative Diabetic Retinopathy by Optical Coherence Tomography Angiography.

PURPOSE: To classify retinal neovascularization in untreated early stages of proliferative diabetic retinopathy (PDR) based on optical coherence tomography angiography (OCTA). DESIGN: A cross-sectional study. METHODS: Thirty-five eyes were included. They underwent color fundus photography, fluorescein angiography (FA), and OCTA examinations. Neovascularizations elsewhere (NVEs), neovascularizations at the disc (NVDs), and intraretinal microvascular abnormalities (IRMAs) were scanned by OCTA. The origin and morphology of NVE/NVD/IRMA on OCTA were evaluated. Retinal nonperfusion areas (NPAs) were measured using ImageJ software. RESULTS: In 35 eyes successfully imaged, 75 NVEs, 35 NVDs, and 12 IRMAs were captured. Three proposed subtypes of NVE were identified based on the origins and morphologic features. Type 1 (32 of 75, 42.67%) originated from the venous side, in a tree-like shape. Type 2 (30 of 75, 40.00%) originated from capillary networks, with an octopus-like appearance. Type 3 (13 of 75, 17.33%) originated from the IRMAs, having a sea fan shape. NVD originated from the retinal artery, from the retinal vein, or from the choroid, and arose from the bending vessels near the rim of the optic disc. IRMA originated from and drained into retinal venules, extending into the retina. The initial layer and affiliated NPA were significantly different in the 3 subtypes of NVEs (all P < .01). CONCLUSIONS: OCTA allowed identification of the origins and morphologic patterns of neovascularization in PDR. The new classification of retinal neovascularization may be useful to better understand pathophysiological mechanisms and to guide efficient therapeutic strategies.

EFFECT OF BODY POSITION ON INTRAOCULAR PRESSURE IN SILICONE OIL TAMPONADE EYES.

PURPOSE: To investigate the effect of body position on intraocular pressure (IOP) in silicone oil tamponade eyes. METHODS: This prospective study included 18 eyes from 18 silicone oil tamponade patients and 24 eyes from 24 healthy subjects. Intraocular pressures were measured by Accupen Applanation Tonometer sitting with face forward, sitting with face down, supine, nondependent lateral decubitus, dependent lateral decubitus, and prone positions. The IOPs in each position and the magnitudes of IOP change were compared between the silicone oil and normal groups. RESULTS: In both groups, the IOPs in sitting positions were significantly lower than that of each recumbent position. The IOPs were highest in prone among all positions. No significant difference was found between IOPs of each group in each position. Between both groups, the IOP elevations in each position had no statistical difference compared with sitting with face forward. CONCLUSION: The IOP is lowest in the sitting position and highest in the prone position in both silicone oil and normal groups. Between both groups, the amount of IOP elevations is equivalent in each position compared with sitting with face forward. Ophthalmologists should be aware that IOP is higher in the prone position and that it should be monitored accordingly.