Proteomic study of aqueous humour in diabetic patients with cataracts by TMT combined with HPLC-MS/MS.

BACKGROUND: The purpose of this study is to identify the proteomic differences between the aqueous humour of diabetes patients with cataracts and that of non-diabetic sufferers of cataracts in a clinical setting. METHODS: Patients were divided into the diabetic experimental group and the non-diabetic control group. Aqueous humour specimens were obtained via cataract surgery. Sample proteins were treated with a TMT reagent, separated using a cation chromatography column, and analysed using a C18 desalting column. Proteins were identified using HPLC-MS/MS. The differential proteins were identified using both a p value of < 0.05 and a fold change of > 1.2. GO classification enrichment analysis, KEGG pathway enrichment analysis, protein interaction network analysis, and ingenuity pathway analysis were all carried out. The expression level of four differential proteins were verified by Western blot, and GC and TTR expressions were further examined using an expanded sample pool. RESULTS: The postprandial glucose levels between the experimental group (9.40 ± 1.35 mmol/L) and the control group (6.56 ± 0.81 mmol/L) were significantly different, with a p value of 1.16E-06. It is important to note, however, that the baseline levels of the parameters showed no statistical differences. In total, 397 aqueous humour proteins were identified; of these, 137 showed significant differences, with 63 upregulated ones and 74 down-regulated ones. The differential proteins play important roles in numerous biological processes and pathways, such as complement and coagulation cascades (p = 1.71E-09). Some of these differential proteins are associated with diabetic retinal degeneration and other diabetic complications. Differential proteins, such as HP, GC, and TTR, have high node degree in the protein interaction network. Western blot results further confirmed that GC were down-regulated while TTR was up-regulated in aqueous humour under diabetic condition. CONCLUSION: A list of differential proteins in the human aqueous humour of diabetic patients was established. Proteins with high interaction scores as per protein interaction analysis, such as GC and TTR, were further verified and could potentially be used as early diagnostic markers for diabetic eye complications in clinical practice.

Hydroxycamptothecin regulates scar formation of the filtration channel under scleral flap by inhibiting the proliferation of scleral fibroblasts.

BACKGROUND: To investigate the inhibitory effect of a hyaluronic acid hydrogel loaded with hydroxycamptothecin (HCPT) on scar formation after filtration surgery in a rabbit model. METHODS: Scleral fibroblasts were isolated and extracted from rabbits' eyes. After treatment with different concentrations of HCPT, cytotoxicity was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and proliferation and extent of apoptosis were analysed using flow cytometry. Hydrogels loaded with different dosages of HCPT were prepared and placed under the scleral flap after the filtration surgery. One day, one week, and two weeks after surgery, follicular, conjunctival, corneal, and anterior chamber inflammation and iris and lens changes were observed. RESULTS: In vitro, compared with cells not treated with HCPT, cells treated with HCPT had decreased survival rate and proliferation, and the apoptosis level increased with increasing HCPT concentrations (p < 0.05). In vivo, the flattening time of filtering blebs in the three groups treated with different dosages of HCPT hydrogel was delayed. The degrees of oedema, inflammation, and bleeding were similar to those observed in the control group. The HCPT hydrogel effectively downregulated the expression of collagen 1 and 3 and tissue inhibitor of metalloproteinase 2 and upregulated the expression of matrix metalloproteinase 2 in a dose-dependent manner. CONCLUSIONS: HCPT significantly inhibited the growth of rabbits' scleral fibroblasts and effectively inhibited scar formation after filtering surgery by accelerating the degradation of extracellular matrix deposition.

Identification of miRNA-mRNA Regulatory Networks Associated with Diabetic Retinopathy using Bioinformatics Analysis.

INTRODUCTION: Diabetic retinopathy (DR) is a major complication of diabetes and a leading cause of visual loss. This study aimed to explore biomarkers for DR that may provide additional reference to DR pathogenesis and development. METHODS: The differentially expressed genes (DEGs) between the DR and control samples in the GSE53257 dataset were identified. Logistics analyses were performed to identify DR-associated miRNAs and genes, and correlation analysis was performed to determine the correlation between them in GSE160306. RESULTS: A total of 114 DEGs in DR were identified in GSE53257. Three genes, including ATP5A1 (down), DAUFV2 (down), and OXA1L (down), were differentially expressed between DR and control samples in GSE160306. Univariate logistics analysis identified that ATP5A1 (OR=0.007, p = 1.40E-02), NDUFV2 (OR = 0.003, p = 6.40E-03), and OXA1L (OR = 0.093, p = 3.08E-02) were DR-associated genes. ATP5A1 and OXA1L were regulated by multiple miRNAs, of which hsa-let- 7b-5p (OR = 26.071, p = 4.40E-03) and hsa-miR-31-5p (OR = 4.188, p = 5.09E-02) were related to DR. ATP5A1 and OXA1L were closely correlated with each other in DR. CONCLUSION: The hsa-miR-31-5p-ATP5A1 and hsa-let-7b-5p-OXA1L axes might play novel and important roles in the pathogenesis and development of DR.

Modified suture-assisted canaloplasty in Asians with primary open-angle glaucoma: a prospective study with 12-month follow-up.

BACKGROUND: To evaluate the efficacy and safety of modified suture-assisted canaloplasty in Asians with primary open-angle glaucoma (POAG). METHODS: A prospective, consecutive cases study, evaluating a modified canaloplasty by twisted 6/0 suture was performed on Asian POAG patients. Three modifications of this canaloplasty included opening the Schlemm's canal by viscocanalostomy, circumferential probing by a twisted 6/0 suture and loose suture of the superficial scleral flap. The twisted 6/0 suture was selected as a prober based on characteristical analysis of size and contact measurement as well as chemical composition conducted among 5/0, twisted 6/0 polypropylene sutures and the microcatheter. Success criteria were defined as intraocular pressure (IOP) ≤ 21 mmHg, 18 mmHg, 15 mmHg, and ≥ 20% reduction without (complete success) or with medications (qualified success). Efficacy was assessed by the success rate of circumferential catheterization, IOP values, the success rate of the surgery, the number of IOP-lowering medications, best-corrected vision acuity (BCVA), cup-to-disc ratio (C/D), and mean deviation (MD). Safety was evaluated by adverse events. RESULTS: Forty eyes from 40 consecutive patients were included with a mean follow-up of 14.8 ± 3.0 months. Circumferential catheterization was successfully conducted in 36 eyes (90%). Mean IOP decreased from 26.2 ± 6.9 mmHg to 14.5 ± 2.7 mmHg at 12 months postoperatively. While medication numbers were reduced from 3.2 ± 0.6 to 0.5 ± 0.8 at month 12 (both p < 0.001). Qualified success rate was 97.2% [95% confidence interval (CI) 0.92-1.03], 86.1% (95% CI 0.74-0.98) and 66.7% (95% CI 0.51-0.83) at 12 months with three criteria. BCVA, C/D and MD did not show progression at 1-year follow-up (p > 0.05). Age, baseline IOP, and spherical equivalent negatively influenced the success rate significantly (all p < 0.05). Adverse events included hyphema (30.6%), IOP spike > 25 mmHg (8.3%), and peripheral synechia to the trabecular-Descemet's membrane (2.7%). CONCLUSION: Twisted 6/0 suture can be an ideal material for cannulation. Modified suture-assisted canaloplasty is an effective, safe alternative with a cost-efficient feature for patients with POAG, especially in developing countries. TRIAL REGISTRATION: This trial was registered in the Chinese Clinical Trial Registry ( ChiCTR1900028618 , 29/12/2019).

Comparison of viscocanalostomy plus suture-assisted near-360-degree trabeculotomy and viscocanalostomy plus rigid probe trabeculotomy in primary congenital glaucoma.

PURPOSE: To compare the efficacy and safety of viscocanalostomy plus near-360-degree suture trabeculotomy (VST) with viscocanalostomy plus rigid probe trabeculotomy (VT) in treating primary congenital glaucoma (PCG) over a one-year follow-up. METHODS: This consecutive retrospective study included patients with PCG confirmed within 3 years of age from March 2017 to October 2019. Efficacy was evaluated by comparing the postoperative intraocular pressure (IOP) curve and the success rate at one year after surgery. Safety was assessed by comparing the postoperative complications. The number of anti-glaucoma agents, horizontal corneal diameter (HCD) and cup-to-disc ratio (C/D) of the two surgical methods were also compared. RESULTS: Data of 90 eyes from 61 patients were analysed. The baseline parameters of the two groups were similar. The IOP at 12 months after surgery in the VST group was 12.7 ± 4.8 mmHg, while that in the VT group was 15.8 ± 6.5 mmHg. The IOP at 6, 9 and 12 months postoperatively in the VST group was significantly lower than in the VT group (p < 0.05). Viscocanalostomy plus near-360-degree suture trabeculotomy (VST) remained a significant favourable factor for complete one-year success (93.6% versus 74.4%, p = 0.005) but not qualified one-year success (97.9% versus 88.4%, p = 0.06). The number of anti-glaucoma agents, HCD and C/D were reduced in both groups. Postoperative complications were not significantly different between the two groups. CONCLUSIONS: In children with PCG, VST provides a more durable IOP control than VT over the one-year follow-up, with a similar safety profile.

Mutational analysis of mitochondrial tRNA genes in 138 patients with Leber's hereditary optic neuropathy.

INTRODUCTION: Mutations in mitochondrial DNA (mtDNA) are the most important causes for Leber's hereditary optic neuropathy (LHON). Of these, three primary mtDNA mutations account for more than 90% cases of this disease. However, to date, little is known regarding the relationship between mitochondrial tRNA (mt-tRNA) variants and LHON. AIM: In this study, we aimed to investigate the association between mt-tRNA variants and LHON. METHODOLOGY: One hundred thirty-eight LHON patients lacking three primary mutations (ND1 3460G > A, ND4 11778Gxs > A, and ND6 14484 T > C), as well as 266 controls were enrolled in this study. PCR-Sanger sequencing was performed to screen the mt-tRNA variants. Moreover, the phylogenetic analysis, pathogenicity scoring system, as well as mitochondrial functions were performed. RESULTS: We identified 8 possible pathogenic variants: tRNAPhe 593 T > C, tRNALeu(UUR) 3275C > T, tRNAGln 4363 T > C, tRNAMet 4435A > G, tRNAAla 5587 T > C, tRNAGlu 14693A > G, tRNAThr 15927G > A, and 15951A > G, which may change the structural and functional impact on the corresponding tRNAs, and subsequently lead to a failure in tRNA metabolism. Furthermore, significant reductions in mitochondrial ATP and MMP levels and an overproduction of ROS were observed in cybrid cells containing these mt-tRNA variants, suggesting that these variants may lead to mitochondrial dysfunction which was responsible for LHON. CONCLUSION: Our study indicated that mt-tRNA variants were associated with LHON, and screening for mt-tRNA variants were recommended for early detection, diagnosis, and prevention of maternally inherited LHON.

The clinical and genetic findings in a Chinese family with Axenfeld-Rieger syndrome.

Objecive: To describe the clinical and genetic findings of an Axenfeld-Rieger syndrome (ARS) family with a new PITX2 splicing mutation. Methods: A Chinese ARS family with five affected individuals was recruited. Exome sequencing was performed on the proband and the variant (C.253-9C > A) in PITX2 gene was detected as a pathogenic mutation. Sanger sequencing was performed for verification and cosegregation analysis. Real-time polymerase chain reaction (RT- PCR) and Western blotting were performed to verify the expression of the pathogenic gene. Results: All the patients showed abnormalities in the anterior segment of both eyes including posterior embryotoxon, corectopia, iris dysplasia, and iridocorneal tissue adhesions. In addition, they all presented systemic features, including maxillary hypoplasia, underbite, hypodontia, conical teeth. Only III-7 showed obvious umbilical skin. In the PITX2 family, we identified a novel heterozygous splicing mutation (C.253-9C > A) which was confirmed by Sanger sequencing to be completely cosegregated with the ARS phenotype. Real-time quantitative PCR and Western results showed that PITX2 mRNA and protein expression were significantly lower in patients compared with unrelated normal controls. Conclusion: In the ARS pedigree, we summarized the variable phenotype, described a novel PITX2 splicing mutation which expand the genetic spectrum of ARS. We further confirmed the possibility of development of ARS induced by this PITX2 gene deficiency.

Pembrolizumab for choroidal metastases from non-small cell lung cancer: a case report and literature review.

We described a 65-year-old male with choroidal metastases (CM) from non-small cell lung cancer (NSCLC). Pembrolizumab (Keytruda) combined with pemetrexed and capecitabine achieved excellent outcomes. After two cycles of pembrolizumab and chemotherapy, blurred vision and left eye pain were significantly relieved. Imaging and ophthalmologic examinations demonstrated complete resolution of the CM, as well as reduction of pulmonary shadow. CM from NSCLC shows complete and durable response to pembrolizumab and chemotherapy. We suggesting that immunotherapy combined with chemotherapy is a promising treatment for CM from NSCLC.

Comparison of ultrasound cycloplasty and transscleral cyclophotocoagulation for refractory glaucoma in Chinese population.

BACKGROUND: To compare the efficacy and safety of focused ultrasound cycloplasty (UCP) and transscleral cyclophotocoagulation (TSCP) in the treatment of refractory glaucoma in a Chinese population. METHODS: We retrospectively compared twenty-eight eligible patients with refractory glaucoma, who were divided into the UCP group and TSCP group. Patients in these two groups underwent a corresponding procedure from June 2018 to February 2019. The intraocular pressure (IOP), visual acuity, the number of anti-glaucoma agents used and complications were reviewed and compared between groups. Proper statistical methods were selected according to comparison models under IBM SPSS 25 software. RESULTS: After the 12-months follow-up, postoperative IOP and number of anti-glaucoma agents used in the two groups were both reduced than the baseline level, and the differences were statistically significant (P < 0.05). There were no significant differences in IOP, number of anti-glaucoma agents and the best-corrected visual acuity between the two groups at each follow-up time point (P>0.05). In terms of complications, the pain at 1 day after surgery in the UCP group was significantly milder than that in the TSCP group (P < 0.05). And there were no significant differences in other complications between the two groups (P > 0.05). CONCLUSIONS: Both UCP and TSCP are safe and effective methods for the treatment of refractory glaucoma. Nevertheless, pain is less severe after UCP.

Viscocanalostomy combined with nearly 360-degree suture trabeculotomy for the treatment of primary congenital glaucoma: a preliminary report of a novel technique for trabeculotomy.

PURPOSE: The aim of this study was to determine the preliminary efficacy and safety of a novel technique for trabeculotomy for the treatment of primary congenital glaucoma (PCG). METHODS: This retrospective interventional study was conducted on patients with PCG. Pliability test was performed among 5/0 and 6/0 polypropylene sutures and the flexible illuminated microcatheter. Viscocanalostomy was first performed and the Schlemm's canal was identified. Then, suture trabeculotomy was completed except the region for viscocanalostomy. The preoperative and follow-up data were recorded and analyzed. Success criteria were defined as intraocular pressure (IOP) ≤ 21 mmHg without (complete success) or with medications (qualified success). RESULTS: 5/0 polypropylene suture was an appropriate option for cannulation of Schlemm's canal. A total of 33 eyes from 23 consecutive patients were included with a mean follow-up of 9.3 ± 3.6 months (range 6-18 months). Circumferential cannulation by suture was successfully performed in 28 eyes (84.8%) of 18 patients. Mean IOP decreased from 33.9 ± 9.4 mmHg preoperatively to 10.5 ± 3.5 mmHg at 6 months (p < 0.001) and 11.3 ± 4.1 mmHg at 9 months (p < 0.001). Complete success rate was 92.9% [95% confidence interval (CI:0.83-1.03)] and 87.5% (CI:0.69-1.06) at 6 months and 9 months, respectively. Hyphema was found in 5 eyes (17.9%), all of which were absorbed within 1 week. No other complications were observed. CONCLUSIONS: Viscocanalostomy combined with nearly 360-degree suture trabeculotomy as a novel technique controls IOP in patients with PCG without any severe complications. It is suitable for extended applications because of accurate identification of Schlemm's canal and low cost.

Efficacy of Ahmed Glaucoma Valve Implantation on Neovascular Glaucoma.

To evaluate the efficacy of Ahmed glaucoma valve (AGV) implantation in treating neovascular glaucoma (NVG) and analyze the factors influencing the surgical success rate, a retrospective investigation of 59 NVG patients (66 eyes) who underwent AGV implantation was conducted at Jiangsu Province Hospital, China, from January 2014 to June 2018. Intraocular pressure (IOP), visual acuity, surgical success rates, medications, and complications were monitored at post-operative 1 day, 1 week, 1, 3, 6 and 12 months. Surgical success criteria were defined as 6 mm Hg < IOP < 21 mmHg with or without additional medications. Results showed average IOP was statistically significant between pre-operative visit and each follow-up visit (all P<0.05). At 12 months, the success rate was 66.7%. Multiple stepwise regression analysis suggested that age, panretinal photocoagulation (PRP), complications and hyphema were significant factors influencing the surgical success rate (all P<0.05). Thus, we conclude that AGV implantation is effective and safe for treatment of NVG. Surgical success is dependent on age, PRP, complications, and hyphema.

Modified viscocanalostomy in the Chinese population with open angle glaucoma: a 10-year follow-up results.

AIM: To study the long-term efficacy and safety of modified viscocanalostomy in Chinese people with open angle glaucoma (OAG). METHODS: This retrospective study included a total of 100 eyes from 100 Chinese patients with medically uncontrolled OAG. All the patients underwent modified viscocanalostomy with injection of viscoelastic material in the surgically created ostia of Schlemm's canal (SC). The modifications included peeling of the inner wall of SC and the juxtacanalicular meshwork, use of mitomycin C, and loosely suturing the superficial scleral flap. Intraocular pressure (IOP), visual acuity, number of medications, laser goniopuncture data and complications were recorded. The definition of complete (qualified) success was an IOP equal to or lower than 21, 18, 16 mm Hg without (with or without) anti-glaucoma medications. RESULTS: The mean IOP was 33.5±9.9 mm Hg before surgery, 15.2±3.6 mm Hg (mean IOP reduction of 51%) at 5y after surgery, and 15.6±2.8 mm Hg (mean IOP reduction of 49.9%) at 10y after surgery (P<0.001). The number of anti-glaucoma medications dropped from 2.39±0.5 preoperatively to 0.47±0.8 at 5y and 0.67±0.8 at 10y postoperatively (P<0.001). The follow-up period was 104.5±37.0mo. The qualified success rate for an IOP of 21, 18 or 16 mm Hg or less was 84% [95% confidence interval (CI): 0.80-0.88], 73% (95%CI: 0.68-0.78), and 59% (95%CI: 0.52-0.66) after 5y, and 80% (95%CI: 0.76-0.84), 69% (95%CI: 0.64-0.74), 51% (95%CI: 0.44-0.58) after 10y, respectively. There was a relationship between age, preoperative IOP and success rate (P<0.01, P<0.05). A total of 31 eyes (31.3%) in 31 patients underwent laser goniopuncture, decreasing the IOP from 22.9±4.3 mm Hg to 16.3±2.5 mm Hg (P<0.01). Neither blebitis nor endophthalmitis occurred. CONCLUSION: Modified viscocanalostomy could be performed to lower IOP, decrease multiple anti-glaucoma drops use as well. It's a safe procedure with less complications over 10y in Chinese individuals with OAG.

Activation of Nrf2/HO-1 pathway protects retinal ganglion cells from a rat chronic ocular hypertension model of glaucoma.

OBJECTIVE: The objective of this work was to find out the effects of nuclear factor erythroid 2-related factor/heme oxygenase-1 (Nrf2/HO-1) pathway on retinal ganglion cell (RGC) injury in glaucoma. METHODS: The chronic ocular hypertension (COH) rat models of glaucoma were constructed, and intraocular pressure (IOP) and RGC numbers were detected at different time points. Additionally, rats were divided into normal group (normal control rats), model group (COH model rats), and model + tBHQ group (COH model rats treated with Nrf activator, tBHQ). RGC apoptosis was detected by using TUNEL staining, and the expressions of Nrf2/HO-1 were detected by qRT-PCR and western blotting. RESULTS: COH model rats showed significant IOP elevation and the increased mRNA and protein expressions of Nrf2 and HO-1 from 1 to 6 weeks after operation, with the evidently decreased RGC numbers at 4 weeks and 6 weeks after operation (all P < 0.05). Besides, rats in the model group had increased apoptosis index (AI) of RGCs and the elevated mRNA and protein expressions of Nrf2/HO-1 with remarkably reduced RGC numbers when compared with normal control rats, but the model rats treated with tBHQ exhibited an apparent decrease in AI of RGCs, as well as remarkable increases in RGC numbers and the mRNA and protein expression of Nrf2/HO-1 (all P < 0.05). CONCLUSION: Activation of Nrf2/HO-1 pathway significantly reduced the apoptosis and injury of RGCs in rats with chronic ocular hypertension (COH), thereby protecting RGCs in glaucoma, which could be a promising clinical target to prevent RGC degeneration in glaucoma.

AAV-mediated human CNGB3 restores cone function in an all-cone mouse model of CNGB3 achromatopsia.

Complete congenital achromatopsia is a devastating hereditary visual disorder. Mutations in the CNGB3 gene account for more than 50% of all known cases of achromatopsia. This work investigated the efficiency of subretinal (SR) delivered AAV8 (Y447, 733F) vector containing a human PR2.1 promoter and a human CNGB3 cDNA in Cngb3 -/-/ Nrl -/- mice. The Cngb3 -/-/ Nrl -/- mouse was a cone-dominant model with Cngb3 channel deficiency, which partially mimicked the all-cone foveal structure of human achromatopsia with CNGB3 mutations. Following SR delivery of the vector, AAV-mediated CNGB3 expression restored cone function which was assessed by the restoration of the cone-mediated electroretinogram (ERG) and immunohistochemistry. This therapeutic rescue resulted in long-term improvement of retinal function with the restoration of cone ERG amplitude. This study demonstrated an AAV-mediated gene therapy in a cone-dominant mouse model using a human gene construct and provided the potential to be utilized in clinical trials.

NKILA inhibition protects retinal pigment epithelium cells from hypoxia by facilitating NFκB activation.

Sustained retinal hypoxia causes injuries to retinal pigment epithelium (RPE) cells. We studied expression and potential functions of nuclear factor-κB (NFκB) Interacting LncRNA (NKILA) in hypoxia-treated RPE cells. Hypoxia induced NKILA expression, NKILA-IκBα association and NFκB activation in ARPE-19 cells and primary human RPE cells. shRNA-mediated knockdown of NKILA facilitated NFκB activation, inhibiting RPE cell death and apoptosis. Conversely, exogenous overexpression of NKILA blocked hypoxia-induced NFκB activation, thereby exacerbating RPE cell apoptosis. Further studies show that hypoxia downregulated microRNA-103 (miR-103), the anti-NKILA microRNA, in RPE cells. Transfection of miR-103 mimic blocked hypoxia-induced NKILA expression to significantly boost NFκB activation, protecting RPE cells from hypoxia. Collectively, we conclude that hypoxia-induced NKILA expression negatively regulates NFκB to promote RPE cell death. Conversely, NKILA inhibition protects RPE cells from hypoxia by facilitating NFκB activation.

Viscocanalostomy combined with trabeculotomy and mitomycin C in the treatment of primary congenital glaucoma.

AIM: To evaluate the long-term outcome of viscocanalostomy combined with trabeculotomy and mitomycin C in the treatment of primary congenital glaucoma. METHODS: This is a retrospective study. Forty-two eyes of 26 patients with primary congenital glaucoma were enrolled. Intraocular pressure (IOP), corneal diameter (mm) and cup/disc (C/D) were measured before and after the surgery respectively. Follow-up period was 30mo. RESULTS: The mean preoperative IOP was 30.6±7.35 mm Hg. Of the 42 eyes, 2 eyes were required conversion to trabeculectomy for the absence of Schlemm's canal. Of remained 40 eyes, 38 eyes (95%) achieved successful IOP control. The average postoperative IOP was 11.69±4.18 mm Hg at 12mo. The mean reduction was 18.91 mm Hg (P<0.0001). Eighteen (75%) eyes presented a reduction in corneal diameter, and 25 (62.5%) eyes presented a C/D ratio reversal after the surgery. There was no serious complication in any patients over the follow-up period. CONCLUSION: Viscocanalostomy combined with trabeculotomy and mitomycin C is useful in the management of primary congenital glaucoma.

Paeoniflorin Suppressed High Glucose-Induced Retinal Microglia MMP-9 Expression and Inflammatory Response via Inhibition of TLR4/NF-κB Pathway Through Upregulation of SOCS3 in Diabetic Retinopathy.

Diabetic retinopathy (DR) is a serious-threatening complication of diabetes and urgently needed to be treated. Evidence has accumulated indicating that microglia inflammation within the retina plays a critical role in DR. Microglial matrix metalloproteinase 9 (MMP-9) has an important role in the destruction of the integrity of the blood-retinal barrier (BRB) associated with the development of DR. MMP-9 was also considered important for regulating inflammatory responses. Paeoniflorin, a monoterpene glucoside, has a potent immunomodulatory effect on microglia. We hypothesized that paeoniflorin could significantly suppress microglial MMP-9 activation induced by high glucose and further relieve DR. BV2 cells were used to investigate the effects and mechanism of paeoniflorin. The activation of MMP-9 was measured by gelatin zymography. Cell signaling was measured by western blot assay and immunofluorescence assay. High glucose increased the activation of MMP-9 in BV2 cells, which was abolished by HMGB1, TLR4, p38 MAPK, and NF-κB inhibition. Phosphorylation of p38 MAPK induced by high glucose was decreased by TLR4 inhibition in BV2 cells. Paeoniflorin induced suppressor of cytokine signaling 3 (SOCS3) expression and reduced MMP-9 activation in BV2 cells. The effect of paeoniflorin on SOCS3 was abolished by the TLR4 inhibitor. In streptozotocin (STZ)-induced diabetes mice, paeoniflorin induced SOCS3 expression and reduced MMP-9 activation. Paeoniflorin suppressed STZ-induced IBA-1 and IL-1ß expression and decreased STZ-induced high blood glucose level. In conclusion, paeoniflorin suppressed high glucose-induced retinal microglia MMP-9 expression and inflammatory response via inhibition of the TLR4/NF-κB pathway through upregulation of SOCS3 in diabetic retinopathy.

Potentially Important MicroRNAs in Form-Deprivation Myopia Revealed by Bioinformatics Analysis of MicroRNA Profiling.

OBJECTIVE: This study aimed to identify differentially expressed microRNAs (miRNAs) and to explore their potential roles in form-deprivation myopia. METHODS: The microarray data set GSE58124 of miRNAs was downloaded from the Gene Expression Omnibus, and form-deprivation myopia was induced in C57BL/6J mice over the right eye; the contralateral, left eyes were used as controls. Differential expression analysis was done using the LIMMA package. miRDB was used to predict targets for miRNAs. The target genes were put into DAVID to identify significant pathways and biological processes of miRNAs. A functionally collaborative network was constructed using Cytoscape. RESULT: In total, 24 and 20 upregulated miRNAs, respectively, were screened out in retina and whole-eye tissue. However, there was no dramatic expression change of miRNAs in sclera tissue. By taking intersections, 8 common upregulated miRNAs were obtained in both the retina and the whole-eye samples. According to miRDB, 1,805 target genes were screened out for the 8 differentially expressed miRNAs, including MAPK10 (mitogen-activated protein kinase 10). The functionally collaborative network revealed that "regulation of transcription" was significantly enriched. The pathways "Axon guidance" and "TGF-ß signaling pathway" were also enriched. Importantly, miR-466h-5p and miR-466j were significantly enriched in some synaptic transmission-related biological processes. CONCLUSION: This study identified an upregulation of 8 miRNAs, which may function by disturbing their enriched pathways or biological processes in the progression of myopia.

Influence of the hypothalamic arcuate nucleus on intraocular pressure and the role of opioid peptides.

BACKGROUND: An opioid peptide neuron/humoral feedback regulation might be involved in changes of intraocular pressure (IOP). The aims of this study are to investigate the effects of arcuate nucleus (ARC) and opioid peptides on intraocular pressure (IOP). METHODS: Fifty-four healthy purebred New Zealand white rabbits (108 eyes) were randomly divided into 4 groups, including control group, electrical stimulation group, [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO) group, and [D-Pen 2, D-Pen5]- enkephalin (DPDPE) group. Bilateral IOP was measured after unilateral electrical stimulation of the ARC or unilateral microinjection into the ARC of the selective µ-opioid receptor agonist DAMGO or the selective δ opioid receptor agonist DPDPE, both alone and after pre-administration of either the non-selective opioid receptor antagonist naloxone or saline. RESULTS: Both electrical stimulation in ARC and micro-injection either or opioid receptor agonists, DAMGO or DPDPE, respectively, caused a significant bilateral reduction in IOP (P<0.05) which was more pronounced in the ipsilateral than in the contralateral eye. Pretreatment with naloxone prevented some, but not all IOP reductions. CONCLUSION: The ARC takes part in the negative regulation of IOP, an action that may involve opioid neurons.