Atropine Affects the Outer Retina During Inhibiting Form Deprivation Myopia in Guinea Pigs.

PURPOSE: Atropine has been proven to be effective in retarding myopia progression. However, the underlying mechanism remains unknown. Our purpose was to detect morphological and functional changes caused by atropine during myopic inhibition. METHOD: Twenty 2-week-old guinea pigs were randomly assigned to either the saline group (n = 10) or the atropine group (n = 10). Form-deprived myopia (FDM) and intravitreal injections were applied on the right eyes. The injections were given every 3 days, lasting for 2 weeks. The left eyes served as control. Ocular refraction, axial length, retinal, and choroidal thickness were collected at the start and the end of the experiment. Retinal function was evaluated via full-field electroretinogram (ERG) at the end of treatment. RESULTS: The interocular differences (experimental eye minus control eye) of refraction error (RE), vitreous chamber depth (VCD), and axial length (AL) in the saline group were significantly greater than those in the atropine group (RE, VCD: P < .001, AL: P < .0001). The differences in choroidal thickness between the two groups did not reach statistical significance. However, a decreasing trend of choroidal thickness was observed in the saline group but not in the atropine group. Furthermore, the interocular differences of total retinal and outer retinal thickness in the atropine group were much thicker than in the saline group (P < .001 and P < .01, respectively). The treatment did not affect inner retinal thickness. In photopic ERG, the atropine-treated FDM eyes showed significantly greater a-wave amplitudes compared to the saline group. CONCLUSION: During the process of inhibiting FDM, atropine showed an effect on the outer retina, most likely on the cones, in guinea pigs.

COL10A1 is a novel factor in the development of choroidal neovascularization.

With the dramatic rise in the aging population, researching age-related macular degeneration (AMD), especially the severe form neovascular AMD (nAMD), has become more important than ever. In this study, we found that collagen type X was increased in retina-choroid tissue of mice with laser-induced choroidal neovascularization (CNV) based on immunohistofluorescence. RNA sequencing and bioinformatic analyses were performed to compare the retina-choroid tissue complex of the CNV mouse model to normal controls. Collagen type X alpha 1 chain (Col10a1) was among the most significantly upregulated genes, and the results were validated with an animal model at the mRNA and protein levels by quantitative real-time polymerase chain reaction (qPCR) and western blotting, respectively. COL10A1 was also upregulated in human retinal microvascular endothelial cells (HRMECs), human umbilical vein endothelial cells (HUVECs), RPE19 cells and RF/6A cells under hypoxic conditions. Next, in vitro and in vivo experiments were performed to study the effect of COL10A1 on neovascularization. siRNA knockdown of COL10A1 suppressed the proliferation and tube formation ability of HRMECs under hypoxic conditions. Snail family transcriptional repressor 1 (SNAIL1) and angiopoietin-2 (ANGPT2) were downregulated in COL10A1 knockdown HRMECs under hypoxic conditions and thus were potential downstream genes. Significant decreases in CNV leakage and CNV lesion area, as assessed by fundus fluorescein angiography (FFA) and immunofluorescence of choroidal flat mounts, respectively, were observed in a mouse model intravitreally injected with anti-collagen X monoclonal antibody (mAb) compared to the controls. In conclusion, COL10A1 promotes CNV formation and may represent a new candidate target for the treatment and diagnosis of nAMD and other neovascular diseases.

KATP Opener Attenuates Diabetic-Induced Müller Gliosis and Inflammation by Modulating Kir6.1 in Microglia.

Purpose: This study aimed to determine the effect of pinacidil, a nonselective KATP channel opener, on diabetes-induced retinal gliosis and inflammation. Methods: Primary and immortalized cell lines of retinal microglia and Müller cells were used to set up a coculture model. In the trans-well system, microglia were seeded in the upper chamber and Müller cells in the bottom chamber. Microglia were polarized into proinflammatory (M1, with lipopolysaccharide and INF-γ) with or without different pinacidil concentrations before coculturing with Müller cells. The expression of inflammatory or anti-inflammatory genes and protein in microglia, and the expression of glial fibrillary acidic protein (GFAP), Kir4.1, and AQP4 in Müller cells were examined by real-time polymerase chain reaction and Western blot. Pinacidil was injected intravitreally into streptozotocin-induced diabetic rats. Retinal gliosis and inflammation were examined by immunohistochemistry and Western blot. Results: Intravitreal injection of pinacidil alleviated diabetes-induced Müller cell gliosis and microglial activation and reduced vascular endothelial growth factor expression. In vitro study demonstrated that pinacidil inhibited tumor necrosis factor and interleukin-1ß expression in M1-type microglia and alleviated the M1 microglia-induced GFAP expression in the Müller cells. Furthermore, we found that pinacidil on its own, or in combination with IL-4, can upregulate arginase-1 (Arg-1) and Kir6.1 expression in microglial cells. Conclusions: Our results suggest that potassium channels are critically involved in diabetes-induced gliosis and microglial activation. The KATP opener, pinacidil, can reduce microglial activation by upregulating Kir6.1 expression.

Establishment of non-integrate induced pluripotent stem cell line CSUASOi006-A, from urine-derived cells of a PRPF8-related dominant retinitis pigmentosa patient.

Retinitis pigmentosa (RP) is a group of inherited retinal disorders characterized by the progressive photoreceptors and pigment epithelial cells dysfunction. Here, we report the human induced pluripotent stem cell line (iPSC) CSUASOi006-A, generated from urine-derived cells (UCs) of a 17-year-old male patient with clinically diagnosed RP carrying point mutation (c.C5792T) in the pre-mRNA processing factor 8 gene (PRPF8). The newly derived CSUASOi006-A cell line has the patient's same mutation (c.C5792T) and could provide useful resources for studying the pathogenic mechanism of PRPF8-related RP.

Kidney Stone Composition in Third-World Areas: What Kashgar Tells Us?

OBJECTIVE: To analyze urinary stone compositions in patients from Kashgar, China. MATERIALS AND METHODS: We analyzed the components of urinary stones in 732 consecutive patients with urolithiasis admitted to the First and Second People's Hospital of Kashgar Prefecture, Xinjiang, from July 2014 to November 2016. The patients were divided into two groups by ages: group A, 0 to 18 years and group B, >18 years old. The distributions of various stone compositions were analyzed and correlated with the gender and age. RESULTS: The mean age of group A was 3.90 ± 4.09 years and that of group B was 39.88 ± 16.40 years. The overall gender ratio (male:female) was 2.27:1. Ammonium acid urate (AAU) stone was the most frequent stone, male 35.83% and female 33.48%. Female patients were significantly more common than male patients in calcium apatite stone (p = 0.004). Of all 732 cases, patients younger than 18 years were more than patients older than 18 years (58.47% vs 41.53%). The majority of the patients (77.87%) had the stone located in the upper urinary tract. Two peak ages for both genders were noted in 1 to 3 years and 19 to 40 years group of the patients. In group of 1 to 3 years patients, male were more than female (37.60% vs 24.55%, p = 0.001), whereas in the group of 10 to 18 years patients, female were more than male (10.71% vs 4.13%). AAU was the predominant stone component in group <1 year (70. 5%, p < 0.01, as compared with other groups.). Uric acid stone was more prevalent in group >60 years (66.8%, p < 0.01) than in other groups. Patients in 1 to 3 years were in the peak age group of AAU stones in both the upper and lower urinary tract. CONCLUSION: Most of the patients with urolithiasis diagnosed and treated in Kashgar are <18 years old, especially younger than 3 years old. The most frequent stone component in this area was AAU. More than 50% patients <18 years old had AAU stone. The mechanisms that could trigger the high prevalence of AAU stone in patients <18 years old are worth further investigation.

Bisphenol A disrupts glucose transport and neurophysiological role of IR/IRS/AKT/GSK3ß axis in the brain of male mice.

Bisphenol A (BPA), one of the most prevalent chemicals for daily use, was recently reported to disturb the homeostasis of energy metabolism and insulin signaling pathways, which might contribute to the increasing prevalence rate of mild cognitive impairment (MCI). However, the underlying mechanisms are remained poorly understood. Here we studied the effects of low dose BPA on glucose transport and the IR/IRS/AKT/GSK3ß axis in adult male mice to delineate the association between insulin signaling disruption and neurotoxicity mediated by BPA. Mice were treated with subcutaneous injection of 100µg/kg/d BPA or vehicle for 30 days, then the insulin signaling and glucose transporters in the hippocampus and prefrontal cortex were detected by western blot. Our results showed that mice treated with BPA displayed significant decrease of insulin sensitivity, and in glucose transporter 1, 3 (GLUT1, 3) protein levels in mouse brain. Meanwhile, hyperactivation of IR/IRS/AKT/GSK3ß axis was detected in the brain of BPA treated mice. Noteworthily, significant increases of phosphorylated tau and ß-APP were observed in BPA treated mice. These results strongly suggest that BPA exposure significantly disrupts brain insulin signaling and might be considered as a potential risk factor for neurodegenerative diseases.

Insulin signaling disruption in male mice due to perinatal bisphenol A exposure: Role of insulin signaling in the brain.

Bisphenol A (BPA), an environmental estrogenic endocrine disruptor, is widely used for producing polycarbonate plastics and epoxy resins. Available data have shown that perinatal exposure to BPA contributes to peripheral insulin resistance, while in the present study, we aimed to investigate the effects of perinatal BPA exposure on insulin signaling and glucose transport in the cortex of offspring mice. The pregnant mice were administrated either vehicle or BPA (100 µg/kg/day) at three perinatal stages. Stage I: from day 6 of gestation until parturition (P6-PND0 fetus exposure); Stage II: from lactation until delactation (PND0-PND21 newborn exposure) and Stage III: from day 6 of pregnancy until delactation (P6-PND21 fetus and newborn exposure). At 8 months of age for the offspring mice, the insulin signaling pathways and glucose transporters (GLUTs) were detected. Our data indicated that the insulin signaling including insulin, phosphorylated insulin receptor (IR), phosphorylated protein kinase B (p-AKT), phosphorylated glycogen synthase kinase 3ß (p-GSK3ß) and phosphorylated extracellular signal regulated protein kinase (p-ERK) were significantly decreased in the brain. In parallel, GLUTs (GLUT1/3/4) were obviously decreased as well in BPA-treated group in mice brain. Noteworthily, the phosphorylated tau (p-tau) and amyloid precursor protein (APP) were markedly up-regulated in all BPA-treated groups. These results, taken together, suggest the adverse effects of BPA on insulin signaling and GLUTs, which might subsequently contribute to the increment of p-tau and APP in the brain of adult offspring. Therefore, perinatal BPA exposure might be a risk factor for the long-term neurodegenerative changes in offspring male mice.

Effects of Bisphenol A on glucose homeostasis and brain insulin signaling pathways in male mice.

The potential effects of Bisphenol A (BPA) on peripheral insulin resistance have recently gained more attention, however, its functions on brain insulin resistance are still unknown. The aim of the present study was to investigate the effects of BPA on insulin signaling and glucose transport in mouse brain. The male mice were administrated of 100 µg/kg/day BPA or vehicle for 15 days then challenged with glucose and insulin tolerance tests. The insulin levels were detected with radioimmunoassay (RIA), and the insulin signaling pathways were investigated by Western blot. Our results revealed that BPA significantly increased peripheral plasma insulin levels, and decreased the insulin signals including phosphorylated insulin receptor (p-IR), phosphorylated insulin receptor substrate 1 (p-IRS1), phosphorylated protein kinase B (p-AKT), phosphorylated glycogen synthase kinase 3ß (p-GSK3ß) and phosphorylated extracellular regulated protein kinases (p-ERK1/2) in the brain, though insulin expression in both hippocampus and profrontal cortex was increased. In parallel, BPA exposure might contribute to glucose transport disturbance in the brain since the expression of glucose transporters were markedly decreased. In conclusion, BPA exposure perturbs the insulin signaling and glucose transport in the brain, therefore, it might be a risk factor for brain insulin resistance.