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Cell division cycle 42 (CDC42) modulates the pathogenesis of allergic rhinitis (AR) through regulating immunity, allergic response, and T-helper (Th)1/Th2 imbalance. This study aimed to evaluate the potential of CDC42 to reflect disease risk, symptom scores, and Th1/Th2 axis of AR and the correlation of its vertical change with symptom amelioration after treatment.CDC42, Th1 cells, and Th2 cells in the peripheral blood mononuclear cells (PBMCs) and interferon-γ and interleukin-4 in the serum were determined in 200 AR patients. Simultaneously, PBMC CDC42 was detected in 50 non-atopic obstructive snoring patients [as disease controls (DCs)] and 50 healthy controls (HCs).CDC42 was increased in AR patients compared with DCs and HCs (both p 0.001) but showed no difference between DCs and HCs (p = 0.054). In AR patients, CDC42 was positively linked to rhinorrhea, itching, sneezing, and total nasal symptom scores (TNSS) (all p 0.05), but not congestion score (p = 0.052). Meanwhile, CDC42 showed positive correlations with Th2 cells (p 0.001) and interleukin-4 (p = 0.005), a negative correlation with Th1/Th2 axis (p = 0.001), but no correlation with Th1 cells (p = 0.095) or interferon-γ (p = 0.174). Notably, CDC42 at week 4 after treatment (W4) was reduced compared with that at enrollment (W0) (p 0.001) and positively correlated with TNSS at W4 (p 0.001); from W0 to W4, CDC42 change also positively correlated with TNSS change (p = 0.004).CDC42 is elevated and positively correlates with symptom scores and Th2 cells, whose short-term reduction reflects symptom alleviation in AR patients.

Cytomegalovirus (CMV) retinitis is a rare end-organ disease of CMV infection and is a marker of severe immunosuppression, especially in human immunodeficiency virus (HIV)-positive patients. In multiple myeloma (MM) patients, CMV retinitis has been reported in the post-transplant setting, with an incidence lower than 0.2%, and in patients receiving lenalidomide. Here, we describe the first case of CMV retinitis in myeloma patients following B-cell maturation antigen (BCMA)-targeted chimeric antigen receptor T (BCMA CAR-T) cell therapy. In addition to CMV, the patient developed multiple infections including a mouth ulcer, pneumonia, and fungal enteritis. While the complete remission (CR) status of MM was maintained, he regained a visual acuity of 20/1000 after appropriate ophthalmologic treatment. This single case illustrates the potential of BCMA CAR-T therapy to induce profound humoral immunosuppression, and demonstrates an imperative need for an established standard of monitoring and prophylaxis of post-CAR-T infections.

Background: Therapeutic applications of retinal progenitor cells (RPCs) are hindered by their limited proliferation and differentiation capacity and poor ability to migrate into damaged retinal tissue. Our study aimed to explore the effects of HA-CD44 interactions on the regulation of RPCs migration, proliferation and differentiation, and to investigate the underlying regulation mechanisms.Methods: Mouse RPCs were isolated and amplified. Western blot and flow cytometry analyses were used to investigate the expression of CD44 in RPCs. The effects of HA-CD44 interactions on the RPCs behaviors, including migration, proliferation and differentiation, were investigated by MTT assay, CCK8 assay, vertical collagen gel invasion assay, time-lapse imaging, immunocytochemistry, RT-PCR and western blot assay. Furthermore, the downstream signals of HA-CD44 interactions were investigated.Results: CD44 was expressed in RPCs, and HA-CD44 interaction markedly improved RPCs adhesion and migration. The stimulation of miR-21 expression by HA-CD44 interaction was PKC/Nanog-dependent in RPCs. Treatment of RPCs with PKC- or Nanog-specific ASODN or miR-21 antagomir effectively blocked HA-mediated RPCs adhesion and migration. Moreover, ROK/Gab-1 associated PI3K/AKT signaling activation was required in the HA-CD44 interaction mediated RPCs proliferation and neuronal differentiation.Conclusions: Our findings demonstrated new roles for HA-CD44 interaction in regulating both migration, proliferation and neuronal differentiation of RPCs. HA-CD44 signaling could comprise a novel approach to control RPC fates, which may be instructive for the application of RPCs for future therapeutic application.

KNOTTED1-like homeobox (KNOX) transcription factors (TFs) belonging to the homeobox TF family play important roles in plant growth, development, and responses to abiotic and biotic stress. However, little information is available on KNOX TF in pear (

Background To identify the clinical characteristics, prognostic factors and visual outcomes in posterior segment IOFBs patients managed by PPV in a tertiary hospital. Methods A retrospective chart review was performed for 56 patients, who had PPV for IOFBs removal between November 2013 and November 2015. The mechanisms of injury, the nature of the IOFBs, the BCVA before and after the surgery, the penetrating site and the complications of the surgery were all collected. Univariate analyses were conducted to evaluate the prognostic factors. Results The mean age of the patients was 36.4 years. The nature of IOFBs was mainly metal. Most injuries were commonly caused by hammering the metal. The mean preoperative VA was 2.30 logMAR, and mean final VA was 0.92 logMAR. From univariate analysis, good visual outcome was correlated with the good visual acuity before surgery and poor visual outcome was correlated with the macular break and multiple surgeries. Conclusions In a tertiary hospital of eastern China, most cases of IOFBs were work-related. The prognosis of the patients was really well in the patients with good presenting visual acuity. Nevertheless the prognosis was not good for those patients who had macular injury or underwent several surgeries because of retinal detachment, epiretinal membrane or proliferative vitreous retinopathy. Good facilities for eye protection are urgently in demand for the workers indeed.

Purpose: To establish quantitative profile of the morphologic changes among patients with active myopic choroidal neovascularization (mCNV) before and after anti-vascular endothelial growth factor (VEGF) therapy using optical coherence tomography angiography (OCTA) to assess the therapeutic response.Methods: Patients with active mCNV who received anti-VEGF injections between February 2017 to October 2020 and fit the study criteria were retrospectively reviewed. Quantitative analysis of their OCTA images were carried out to evaluate the morphologic features and vascular changes of mCNV lesions in response to anti-VEGF therapy. For further quantitative profiling, mCNV area, fractal dimension, vessel area, vessel density, vessel diameter, vessel length, vessel junction, junction density, and vessel tortuosity were obtained by means of advanced skeletonization postprocessing analyses.Results: Thirty-one eyes of 29 consecutive patients with OCTA-positive mCNV lesions (mean spherical equivalent: −12.55 ± 3.24 diopters) were included. The 31 cases were divided into two phenotypes at baseline: organized interlacing pattern (83.87%) and disorganized vascular loops pattern (16.13%). The values of mCNV area, fractal dimension, vessel area, vessel length, vessel junction, and junction density decreased remarkably 1 month after the initial anti-VEGF injection (p < 0.001). Although, vessel density, vessel diameter, and vessel tortuosity increased meanwhile, only vessel diameter displayed statistical significance (p = 0.027). Of note, relative ratio analysis showed that vessel junction was the most sensitive biomarker in response to anti-VEGF therapy, reflecting a mean decrease of 50.36%. Sensitivity lowered successively in biomarkers of vessel length, vessel area, junction density, mCNV area, and fractal dimension. In addition, percent change of mCNV area (r = 0.552, p = 0.002), fractal dimension (r = 0.446, p = 0.017), vessel area (r = 0.518, p = 0.005), and vessel length (r = 0.440, p = 0.019) were moderately associated with that of central retinal thickness.Conclusions: The study showed morphological as well as quantitative changes on OCTA responding to anti-VEGF treatment in mCNV patients, among which vessel junctions might be the most predictive biomarker. OCTA-based analysis, providing intuitive images and a large spectrum of quantitative data at the same time, could promote new insights into the therapeutic response assessment in mCNV patients.

Background To compare changes in retinal microvasculature of young and elderly patients with retinal vein occlusion (RVO) after anti-VEGF treatment. Methods RVO patients who underwent anti-VEGF treatment were retrospectively reviewed and categorized into two groups based on age. The OCT angiography images were obtained during each visit. Best corrected visual acuity (BCVA), vessel density (VD) and foveal avascular zone (FAZ) were measured and compared between the two groups. Vision improvements and retinal microvasculature changes were also correlated. Results Twenty patients with 20 eyes were enrolled in the younger group and 46 patients with 46 eyes were enrolled in the older group. Younger patients demonstrated better BCVA, higher VD and smaller FAZ than older patients at 12 months after the first anti-VEGF treatment. The improvement of VD was observed only in the younger group. A positive correlation between vision improvement and VD increase was noted. Conclusions Young patients with RVO can achieve rapid rehabilitation of deep retinal vasculature which lead to a better visual outcome.

Purpose. To describe the clinical features, management, and outcomes of patients with posttraumatic endophthalmitis (PTE) and to determine risk factors for poor visual prognosis. Methods. We retrospectively reviewed the medical records of 42 consecutive patients presenting with PTE who were treated at our institution between 2017 and 2019. Each patient’s data, including demographic characteristics, ocular injury details, surgical records, patient outcomes, and laboratory results, were collected and analyzed. Multivariate analysis was conducted to determine the factors associated with poor visual outcomes. Results. In our series, male (n = 36, 85.7%) and patients below 60 years of age (20–40 years, 23.8%; 40–60 years, 57.14%) comprised most of the total cohort. On presentation, 39 (92.8%) of the 42 PTE patients presented best-corrected visual acuity (BCVA) worse than counting fingers. Pars plana vitrectomy (PPV) was performed in all the patients. 59.5% (n = 25) of the patients’ BCVA improved after surgery and 33.3% (n = 14) achieved BCVA of 20/200 or better. The rate of evisceration was 7.1% (n = 3). Of the 42 specimens, the culture was positive in 10 (23.8%) eyes. By univariate analysis, factors including sex, occupation, systemic disease, source of trauma, lens injury, silicone oil tamponade, usage of intravitreal antibiotics, BCVA at presentation, and culture positive for any organism did not affect the final visual outcome. The features associated with poor BCVA (grouped as P = 0.035 ), corneal-sclera wound (versus sclera wound) ( P = 0.047 ), retained intraocular foreign bodies (IOFBs) ( P = 0.006 ), treatment > 3 days (versus P = 0.033 ), and more times of surgeries ( P = 0.033 ). Conclusions. PTE is a severe complication of penetrating globe injuries associated with irreversible visual loss. Our results highlighted the importance of conducting early therapeutic PPV and IOFB removal to achieve better visual outcomes.

Purpose. Siderosis bulbi may occur as a result of retained intralenticular foreign bodies (ILFBs) that were missed during examination in patients with self-sealing wounds and without a significant decrease in visual acuity. This study aimed to explore the clinical characteristics and visual outcomes of ILFBs with self-sealing corneal penetrating wounds. Methods. Fifteen eyes of 15 patients with ILFBs and self-sealing corneal penetrating wounds, seen between October 2014 and September 2019, were retrospectively analyzed. Data regarding the patient demographics, clinical features, surgical procedure, and initial and final best-corrected visual acuity (BCVA) were analyzed. Results. All patients were male with a mean age of 41 years. The foreign bodies passed through the cornea, sometimes through the iris, through the anterior capsule, and finally localized in the lens. All ILFBs were pointed and metallic objects and were successfully removed with phacoemulsification and posterior chamber intraocular lens (IOL) implantation. Anterior capsule violation was found in three eyes, but no posterior capsule rupture was found. The IOL was placed in a capsule bag in all the cases. The BCVA ranged from 20/200 to 20/25 preoperatively and improved to between 20/32 and 20/20 at the last follow-up visit. The IOLs were well-centered. Apart from posterior capsule opacity in four eyes, no other postoperative complications were found. Conclusions. In patients with a pointed metallic ILFB and self-sealing corneal penetrating wounds (with or without cataracts), early diagnosis and removal of the metallic ILFB combined with lens removal and IOL implantation may avoid late complications and achieve good visual outcomes.

Background: Essential fatty acids (EFAs) n3 alpha-linolenic acid (ALA) and n6 linoleic acid (LA) have been shown to improve lipid metabolism, promote nervous system development, suppress inflammation, and are of benefit in diabetes mellitus and cancer. But the exact mechanism of their action(s) is not clear. In the present study, we evaluated the effects of ALA and LA on glucose homeostasis, circulating lipid profile, inflammatory markers, gut microbiome, and bile acids in streptozotocin (STZ)-induced type 1 diabetes with retinopathy in rats. We also measured vitreal LXA4 levels in patients with diabetic retinopathy since arachidonic acid (AA), the precursor of LXA4, is derived from LA. Methods: The study was performed in male Sprague Dawley rats. Type 1 DM was induced by intraperitoneal injection of STZ. The type 1 DM animals received LA or ALA 100 ug/day via intraperitoneal route on alternate days till the end of the study (21 days after the DM onset). In these animals, glucose homeostasis and lipid profile were evaluated. Plasma and retinal fatty acid profile by gas chromatography (GC), retinal histology by H&E staining, activities of desaturases in the retina and liver tissues and inflammatory markers by qRT-PCR, and composition of the gut microbiota by 16S rDNA sequencing technology of the fecal samples were performed. In addition, short-chain fatty acids (SCFAs) and bile acids (BAs) in the feces and liver of rats were assayed by GC and liquid chromatography coupled with tandem mass spectrometry (UHPLC–MS/MS), respectively. Human vitreal samples obtained from patients with proliferative diabetic retinopathy (PDR) were analyzed for their fatty acids profile and LXA4 levels and compared with those with macular hole (control). Findings: LA and ALA-treated STZ-induced type 1 DM animals showed significant reduction in their plasma glucose levels, pro-inflammatory cytokines (IL-6 and TNF-α), changes in the composition of gut microbiome in the form of an increase in the abundance of Ruminococcaceae (ALA-treated group), Alloprevotella, Prevotellaceae_Ga6A1_group, Ruminococcaceae_UCG_010 and Ruminococcus_1 (in LA-treated group) , an increase in acetic acid and butyric acid, and an increase in the fecal concentrations of CA, CDCA and TUDCA and a decrease in GCA. Concentrations of hepatic CDCA, TCDCA, and TUDCA were significantly increased in ALA treated animals while CDCA were elevated in those with LA treatment. STZ-treated animals showed significant decrease in plasma LA, GLA, and AA and ALA levels with no significant changes in retinal fatty acid composition. All these changes reverted to normal in LA and ALA treated group. The changes in the plasma fatty acid composition were accompanied by significant alterations in the expression of desaturases, COX-2 and 5-LOX and 12-LOX enzymes and pro- and anti-inflammatory cytokines. In addition, retinal LXA4 levels were found to be decreased in patients with PDR. Interpretation: The results of the present study suggest that ALA and LA administration can improve plasma glucose levels, suppress inflammation, and produce significant beneficial alterations in the gut microbiota, plasma lipid profile and EFAs metabolism. Decrease in retinal LXA4 levels in those with PDR agree with the suggestion that EFAs metabolism is altered in T1DM and DR. These results imply a significant role for EFAs and their metabolites in the pathobiology of type 1 DM and diabetic retinopathy. Funding Information: The project was funded by the National Natural Science Foundation of China (No. 82000886), and Natural Science Foundation of Zhejiang Province (No.LQ20H120011). Declaration of Interests: Authors have no conflicts of interest to declare. Ethics Approval Statement: This study was approved by the animal ethics committee of the Second Affiliated Hospital of Zhejiang University school of medicine. All experimental procedures using animals were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. This study was approved by the Clinical Ethical Research Committee of the Second Affiliated Hospital of Zhejiang University school of medicine. The study was performed as per the tenets of Helsinki declaration. Informed consent obtained from all the participating patients.

Background: The exact pathogenesis of idiopathic choroidal neovascularization (ICNV) remains unclear. Cytokine-mediated inflammation has been thought to be involved in the pathophysiology of ICNV. The purpose of this study was to investigate serum cytokine profiles in patients with ICNV and to explore the relationship between serum cytokine levels and ICNV severity. Methods: This case-control study was conducted in 32 ICNV patients and 30 healthy volunteers. Clinical and demographic information was obtained from the medical data platform and the serum was analysed with a multiplex assay to determine the levels of seven cytokines: interleukin (IL)-2, IL-10, IL-15, IL-17, basic fibroblast growth factor (basic FGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and vascular endothelial growth factor (VEGF). Results: Serum levels of IL-2, IL-10, IL-17, basic FGF, and VEGF were elevated in ICNV patients compared to controls. Serum GM-CSF levels were positively related to central retinal thickness, and serum IL-17 levels were positively related to CNV lesion area. Conclusion: Serum inflammatory cytokines were significantly elevated in ICNV patients compared to controls. This suggests that systemic inflammation may play a critical role in the physiopathology of ICNV.

Background: There is an increasing trend toward performing pars plana vitrectomy and simultaneous cataract extraction in the management of patients with intraocular foreign bodies and traumatic cataracts. Herein, we describe a case of good visual rehabilitation in a patient without traumatic cataract surgery after removal of a ferrous intravitreal foreign body (IVFB) using an external approach. Case presentation: A 44-year-old man presented to our ophthalmology department because of decreased visual acuity after a penetrating trauma to the left eye 2 days earlier. Best-corrected visual acuity (BCVA) was 20/200. An examination revealed a midperipheral self-sealing corneal penetrating wound at the 5 o’clock position, in addition to a mild inflammatory reaction in the anterior chamber. Pupil dilation revealed peripheral anterior and posterior capsular violations, with subcapsular opacity involving the visual axis. A small metallic-like foreign body suspended in the vitreous was confirmed by B-scan ultrasonography and orbital computed tomography. Retinal injury and vitreous hemorrhage were excluded by B-scan ultrasonography and binocular indirect ophthalmoscopy. This IVFB was successfully removed by external magnetic extraction through a pars plana incision. At the 2-week follow-up, the traumatic cataract was partially resolved, and BCVA improved to 20/125. At the 3-month follow-up, the traumatic cataract was mostly resolved, the visual axis was clear under a normal pupil, and BCVA improved to 20/20. Peripheral localized lens opacity was found after pupil dilation, and a fundus examination revealed no obvious abnormality. Conclusions: The size and location of an intraocular foreign body and the location and extent of lenticular involvement and associated injuries should be considered in patients with intraocular foreign bodies and traumatic cataracts. In selected patients with a small ferrous IVFB and a localized traumatic cataract, a good visual outcome may be achieved with minimal surgery. Keywords: Intravitreal foreign body, Traumatic cataract, External approach, Case report.

Twelve ingredients obtained from new, non-genetically modified soybean cultivars as well as commercially available sources of soybean meal (SBM) were evaluated in practical feed formulations for the Pacific white shrimp, Litopenaeus vannamei , using both digestibility and growth trials. Apparent digestibility coefficients for energy (ADE) and protein (ADP) were determined using chromic oxide as inert marker and the 70:30 replacement technique. Digestibility coefficients for ADE and ADP ranged from 76.6 to 91.3% and 93.6 to 99.8%, respectively. In general, two distinct groups of high and moderate digestibility values were observed. A separate 8-week growth trial was conducted to evaluate selected SBMs using six replicate tanks per dietary treatment (10 shrimp per tank, initial weight 0.52 ± 0.04 g). An SBM-based reference diet was formulated using commercial SBM (45.3), which was then completely replaced on iso-nitrogenous basis with other experimental SBMs. Significant differences ( P ≤ 0.05) were observed in growth performance but not survival. In general, trends in digestibility values were similar to those of the growth trial. Ingredients (I)-16, I-17, I-18, and I-19 demonstrate relatively higher values in terms of digestibility as well as supporting good growth. Results indicate that genetic selection of soybean for nutritional characteristics of the meal has the potential to increase the nutritional values of SBM for shrimp feeds. Statement of relevance This study provides useful and important information of 12 varieties soybean meal in shrimp to support the aquaculture feed industry development.

Use of distillers dried grain with solubles from sorghum (sDDGS) was studied with respect to processing and physicochemical quality of shrimp feed, followed by growth trials with Litopenaeus vannamei (Pacific white shrimp). Shrimp diets with 0%, 10%, 20%, 30% and 40% sDDGS inclusion, as a replacement for soybean meal, were produced using extrusion and steam pelleting. Bulk density of extruded feed (0.53–0.58 g cm−3) was lower than that of pelleted feed (0.61–0.65 g cm−3), although sDDGS level did not have an impact. Finished diets were 100% sinking, with some exceptions in the case of extruded feed. Pellet durability index (89.4–96.3%) had an increasing trend up to 20% and 30% sDDGS for extruded and pelleted diets respectively. Extruded feed had higher degree of gelatinization than pelleted feed, although proportion of gelatinized starch generally decreased with sDDGS level. Water stability (76.2–91.6%) was higher for extruded feed as compared to pelleted feed, and remained unchanged or decreased with sDDGS level. The extruded and pelleted diets were evaluated in two growth trials with L. vannamei for duration of 9 and 6 weeks in 40 and 60 tanks (initial weight 0.36–0.38 g; 10 shrimps per tank) respectively. Significant differences were not observed in final mean weight and survival with respect to sDDGS level, indicating that up to 40% of this novel protein source can be used in feed formulations without affecting the performance of L. vannamei. Comparison of extruded feed with pelleted feed for impact on mean weight and feed conversion ratio did not yield conclusive results.

Heat shock proteins (HSPs) were originally identified as stress-responsive proteins and serve as molecular chaperones in different intracellular compartments. Translocation of HSPs to the cell surface and release of HSPs into the extracellular space have been observed during the apoptotic process and in response to a variety of cellular stress. Here, we report that UV irradiation and cisplatin treatment rapidly induce the expression of membrane-bound Hsp60, Hsp70, and Hsp90 upstream the phosphatidylserine exposure. Membrane-bound Hsp60, Hsp70 and Hsp90 could promote the release of IL-6 and IL-1β as well as DC maturation by the evaluation of CD80 and CD86 expression. On the other hand, Hsp60, Hsp70 and Hsp90 on cells could facilitate the uptake of dying cells by bone marrow-derived dendritic cells. Lectin-like oxidized LDL receptor-1 (LOX-1), as a common receptor for Hsp60, Hsp70, and Hsp90, is response for their recognition and mediates the uptake of dying cells. Furthermore, membrane-bound Hsp60, Hsp70 and Hsp90 could promote the cross-presentation of OVA antigen from E.G7 cells and inhibition of the uptake of dying cells by LOX-1 decreases the cross-presentation of cellular antigen. Therefore, the rapid exposure of HSPs on dying cells at the early stage allows for the recognition by and confers an activation signal to the immune system.

text, 紀要論文 / Departmental Bulletin Paper

text, 紀要論文 / Departmental Bulletin Paper

The Escherichia coli homolog of GroEL, a 60 kDa heat shock protein (HSP), is a dominant protein produced not only in response to heat stress but also under in vitro growth condition. Beside its traditional cytoplasmic location, the surface exposures of GroEL have been observed in many pathogenic bacteria. To investigate the role of the surface-associated GroEL in the binding of E. coli to macrophages, we constructed a new strain of E. coli displaying GroEL on the outer membrane. We found that surface-associated GroEL increases the clearance ratio of E. coli by macrophages. It has been previously demonstrated that lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is the receptor for Hsp60 from different species. Our present results showed that GroEL on E. coli was recognized by LOX-1 on macrophages, leading to the phagocytosis of pathogen by macrophages. In addition, surface-associated GroEL made mice more susceptible to E. coli-induced peritonitis. These findings add to the research that clarifies the factors mediating bacterial adherence to host cells. Our results suggest that GroEL is a novel therapeutic target for modulating the immune response in infectious and inflammatory conditions.

Idiopathic choroidal neovascularization (ICNV) is a disorder that primarily affecting patients younger than 50 years and can cause severe loss of vision. Choroidal abnormalities, especially choroidal inflammation, have been thought to be involved in the pathophysiology of ICNV. However, the exact pathogenesis of ICNV remains unclear. The aim of our study was investigate the levels of 27 inflammatory cytokines in the aqueous humor of eyes with ICNV and to determine the effect of intravitreal injection of ranibizumab (IVR) on cytokine levels. Significantly higher levels of IL-2, IL-10, IL-15, IL-17, basic FGF and GM-CSF were observed in patients with ICNV compared with controls. However, only IL-17 levels were significantly higher in patients with ICNV compared with controls after adjusting for axial length. Furthermore, there were significant correlations between the levels of IL-10, IL-17, GM-CSF and VEGF and the lesion area. Significant changes in visual acuity and central retinal thickness were observed after IVR. Besides VEGF, IVR also significantly reduced the levels of IL-2, IL-10, basic FGF and IL-12, however, the IL-6 levels were significantly increased. Our results suggest that there may be an involvement of IL-17-related inflammatory processes in the etiology of ICNV.

Microglial activation plays a crucial role in the pathological processes of various retinal and optic nerve diseases. TNF-α is a pro-inflammatory cytokine that is rapidly upregulated and promotes retinal ganglion cells (RGCs) death after optic nerve injury. However, the cellular source of TNF-α after optic nerve injury remains unclear. Thus, we aimed to determine the changes of retinal microglial activation in a rat model of optic nerve transection (ONT) after transcorneal electrical stimulation (TES). Furthermore, we assessed TNF-α expression after ONT and evaluated the effects of TES on TNF-α production. Rats were divided into 2 control groups receiving a sham surgery procedure, 2 ONT+Sham TES groups, and 2 ONT+TES groups. The rats were sacrificed on day 7 or 14 after ONT. RGCs were retrogradely labelled by Fluorogold (FG) 7 days before ONT, one TES group and corresponding controls were stimulated on day 0, 4, and the second were stimulated on day 0, 4, 7, 10. Whole-mount immunohistofluorescence, quantification of RGCs and microglia, and western blot analysis were performed on day 7 and 14 after ONT. TES significantly increased RGCs survival on day 7 and 14 after ONT, which was accompanied by reduced microglia on day 7, but not 14. TNF-α was co-localized with ameboid microglia and significantly increased on day 7 and 14 after ONT. TES significantly reduced TNF-α production on day 7 and 14 after ONT. Our study demonstrated that TES promotes RGCs survival after ONT accompanied by reduced microglial activation and microglia-derived TNF-α production.

Purpose. To investigate the prevalence and characteristics of focal choroidal excavation (FCE) concurrent with central serous chorioretinopathy (CSC) using multimodal imaging.Methods. This was a retrospective single-institution study. Clinical features and multimodal imaging findings were analyzed in eyes with CSC and FCEs, using imaging methods including optical coherence tomography (OCT), OCT angiography (OCTA), fluorescein angiography (FA), indocyanine green angiography (ICGA), fundus autofluorescence (FAF), and multispectral imaging.Results. Seventeen patients (4.8%) with 21 FCEs (19 eyes) were found among 351 consecutive Chinese patients with CSC. Chronic CSC represented 47.1% of those cases. Window defects in 12 lesions identified through FA and hypoautofluorescence in 13 lesions identified through FAF revealed retinal pigment epithelial attenuation. Choroidal hemodynamic disturbances characterized by localized filling defects at the excavation and circumferential hyperperfusion were validated by both ICGA and OCTA, which were similar to the angiographic features of normal chronic CSC. The hyperreflective tissue beneath FCE, observed on B-scan OCT, presented as intensive choroidal flow signals on OCTA.Conclusions. FCE is not uncommon in patients with CSC. Multimodal imaging suggested that the aberrant choroidal circulation might be a contribution factor for leakage from the dysfunctional retinal pigment epithelium at the area of excavation.

Objective and design To determine whether the neurokinin-1 receptor (NK1R) plays a role in the activation of RBL-2H3 mast cells after FcεRΙ aggregation. Materials and methods NK1R expression in RBL-2H3 cells was inhibited by small hairpin RNA (shRNA) against NK1R, and determined by western blotting. For activation, both NK1R knockdown and control RBL-2H3 cells were sensitized by dinitrophenol (DNP)-specific IgE and stimulated with the antigen DNP-bovine serum albumin (BSA). Following the activation of RBL-2H3 cells, monocyte chemoattractant protein (MCP-1) production and intracellular calcium flux were monitored by ELISA and confocal microscopy assay, respectively. For investigation of the signaling mechanism, phosphorylation of mitogen-activated protein kinases (MAPKs) after RBL-2H3 cell activation was assessed by western blotting. Results shRNA-NK1R mediated an effective inhibition of NK1R expression in RBL-2H3 cells. Protein production of MCP-1 was reduced by more than 55 % in NK1R knockdown RBL-2H3 cells compared with control RBL-2H3 cells. In addition, both calcium mobilization and phosphorylation levels of MAPKs (Erk1/2, JNK, and p38) after DNP-BSA stimulation (via FcεRΙ) were decreased due to the inhibition of NK1R expression. Conclusion NK1R is required for the activation of RBL-2H3 cells following FcεRΙ engagement and involved in the regulation of MAPK signaling pathways. Electronic supplementary material The online version of this article (doi:10.1007/s00011-012-0523-x) contains supplementary material, which is available to authorized users.

To study RhoA activation and the effect of the Rho-kinase inhibitor in the development of retinal neovascularization in a mouse model of oxygen-induced retinopathy (OIR).C57BL/6 mice at postnatal day (P7) were exposed to hyper-oxygen for 5 days and returned to room air for 5 days to induce OIR. RhoA-GTP, an active form of RhoA, in retinas at P12, P13 and P17 was detected. Mice received a single intravitreal injection of Y27632 (5 µM or 50 µM), a Rho-kinase inhibitor, in one eye during the transition from oxygen to room air at P12. Contralateral eyes were used as the control. Fluorescein-conjugated dextran angiography of retinal whole mount was prepared to score features of neovascular retinopathy at P17. The preretinal neovascular nuclei quantification was performed in frozen sections as well to evaluate the neovascularization.The retinal RhoA-GTP in OIR mice significantly increased from 0.24 ± 0.06 at P12 to 0.38 ± 0.12 at P13 (p 0.05). The median total retinopathy score of 5.7 was significantly lower in eyes treated with Y27632 than controls (p 0.001). Significant improvement was found in the specific categories of vascular tufts (p 0.01) and extraretinal neovascularization (p 0.05) in treated eyes. Those treated eyes also had a significantly decreased number of neovascular nuclei (p 0.01).These results suggest that Rho/Rho-kinase signaling pathways are involved in the early process of hypoxia-induced retinal neovascularization and Y27632 might have therapeutic potential for the treatment of retinal neovascularization.

PURPOSE To assess the efficacy, duration of effect and safety of one intravitreal injection of bevacizumab in diabetic macular oedema (DMO). METHODS Bevacizumab (1 mg/0.04 ml) was injected intravitreally into eyes with DMO (29 with and nine without previous treatments). Best corrected visual acuity (BCVA), intraocular pressure and central retinal thickness (CRT) were measured; slit-lamp examination, macular biomicroscopy, optical coherence tomography and fluorescein angiography were performed before and at 2-4, 8 and 12 weeks post-injection. Best corrected VA and CRT were analysed in both groups. RESULTS In the non-pretreated group, mean BCVA improved from 0.76 +/- 0.33 (baseline) to 0.57 +/- 0.30 and 0.54 +/- 0.27 at 2-4 weeks and 8 weeks post-injection, respectively (p = 0.02, p = 0.014, paired t-test). Mean CRT decreased from 632.4 +/- 196.0 microm (baseline) to 392.3 +/- 113.6 microm and 370.4 +/- 141.7 microm at the same time-points, respectively (p = 0.01, p = 0.01). There was no difference in BCVA or CRT at 12 weeks. In the pretreated group, mean BCVA improved from 0.62 +/- 0.30 (baseline) to 0.53 +/- 0.33 at 2-4 weeks post-injection (p = 0.01), and mean CRT decreased from 583.9 +/- 180.7 microm (baseline) to 404.1 +/- 197.9 microm at 2-4 weeks post-injection (p < 0.001). Mean BCVA was unchanged at 8 weeks and 12 weeks post-injection, although mean CRT remained lower at 8 weeks (p = 0.004). No ocular or systemic side-effects developed during follow-up. CONCLUSIONS One intravitreal injection of bevacizumab for DMO seems to be effective and safe in both eyes that have been treated previously and eyes that have not. The therapeutic effect is temporary and repeat treatment may be needed.

To determine the effectiveness of staining an epiretinal membrane (ERM) with triamicinolone acedonide (TA) during vitrectomy for a retinal detachment (RD) due to a macular hole (MH) in highly myopic eyes.Thirty-four highly myopic eyes (-6.0 dioptres) of 34 patients with an RD caused by an MH underwent vitrectomy with TA-assisted ERM removal. The excised ERMs from five eyes were examined by transmission electron microscopy (TEM). The main outcome measures were the retinal status and best-corrected visual acuity (BCVA). The anatomic status of the MH was determined by optic coherence tomography (OCT) in 24 successfully retinal-reattached eyes. The follow-up periods ranged from 5 to 20 months.The granules of TA adhered to the ERM and the residual posterior hyaloid, which made them more visible and facilitated their removal. Tissues resembling the internal limiting membrane were not detected in any of the five excised tissues. The retinal reattachment rate was 88% after the first surgery. The mean log MAR BCVA significantly improved from 1.83+/-0.50 (mean+/-SD) before surgery to 1.22+/-0.35 at the final follow-up (P0.001, Wilcoxon signed rank test). OCT showed a normal concave appearance in 11 of 24 eyes (46%), but a central neural epithelial defect was present in 13 of 24 eyes (54%).TA facilitates the complete removal of both the ERM and residual posterior hyaloid, thus ensuring high anatomical and visual success rates in highly myopic eyes with an RD due to MH.

To determine the safety and efficacy of transsclerally placed intrapapillary wire microelectrodes implanted chronically into the optic nerve head of rabbit eyes.Four platinum wire microelectrodes were passed through the sclera and implanted into the optic nerve head of five rabbit eyes for 4-6 months. Color fundus photography, fluorescein angiography, electroretinograms (ERGs), and visually evoked potentials (VEPs) were used to monitor the retina. Electrically evoked potentials (EEPs) were elicited by bipolar electrical stimulation of the optic nerve axons by different combinations of the four electrodes immediately after the implantation and at 1-month intervals thereafter. The effects of the chronic implantation of the electrodes on the morphology of the optic nerve were evaluated by histological and immunohistochemical examinations at 4 and 6 months after the implantation.All of the electrodes remained stable in the implanted sites throughout the post-implantation period, except for one electrode that had pulled out of the optic nerve head at 1 month after implantation. No intraocular infection, inflammation, or vitreoretinal proliferation was observed in any eye. EEPs could be elicited from each pair of electrodes at all testing times. The mean threshold currents (charge densities) to evoke EEPs increased from 19.3 +/- 9.2 microA (6.0 +/- 2.9 microC/cm2) on the implantation day to 78.8 +/- 31.9 microA (24.6 +/- 10.0 microC/cm2) at 1 month after implantation, but did not change significantly thereafter. The implicit time and amplitude of the a- and b-waves of the ERGs and of P1 of the VEPs did not change significantly throughout the post-implantation period. Histological evaluation of the optic nerve head revealed slight tissue encapsulations surrounding the electrode and increased expression of glial fibrillary acidic protein near the surface of the optic nerve.Implantation of transscleral intrapapillary microelectrodes appears to be safe and effective. These findings indicate that the implantation of microelectrodes in the optic nerve head should be considered for an optic nerve-based prosthesis.