Aqueous Humor Of Patients Research Articles

Infrared analysis in the aqueous humor of patients with uveitis: Preliminary results

Inflammatory processes affecting the uvea result in a temporary o permanent blurred vision and represent an important cause of visual impairment worldwide. It is often hard to make a precise diagnosis which is dependent on the clinical expertise, diagnostic tests, laboratory investigations in blood and sometimes in the aqueous humor (AH). With the aim of obtaining proof of principle Fourier Transformed Infrared (FT-IR) absorbance spectroscopy was applied to study the molecular composition of 72 AH samples collected in 26 patients with uveitis and in 44 controls. The unsupervised exploration of the internal structure of the dataset by principal component analysis reduced hundreds IR variables to those most representative allowing to obtain the predictive model that distinguished the AH spectra of patients with uveitis from controls. The same result was obtained by unsupervised agglomerative cluster analysis. After labeling the spectra with some clinical information it was observed that most severe uveitis with active processes were grouped separately from chronic and relapsing uveitis and controls. The consistence of prediction models is discussed in the light of supporting etiological diagnosis by machine learning processes. In conclusion, proof of principle has been obtained that the IR spectral pattern of AH may reflect particular uveal diseases.

Aqueous Humor Cytokine Profile in Primary Congenital Glaucoma.

Cytokine profile in patients with primary open-angle glaucoma (POAG) differs from that in healthy controls. Due to the different pathophysiological mechanisms involved in the genesis of primary congenital glaucoma (PCG) and POAG, it is possible that the cytokine profile could also differ. The main objective of this study was to compare the concentrations of cytokines in the aqueous humor of patients with PCG with those of POAG patients and a control group. A cross-sectional study was conducted. Aqueous humor samples were taken from PCG and POAG patients eligible for glaucoma or cataract surgery and from patients undergoing cataract surgery. Twenty-seven cytokines were analyzed using the Human Cytokine 27-Plex Immunoassay Kit (Bio-Rad Laboratories, Hercules, CA, USA). A total of 107 subjects were included: patients with PCG (n = 19), patients with POAG (n = 54), and a control group (CG) of patients undergoing cataract surgery (n = 34). Most cytokines measured in aqueous humor in PCG presented decreased values compared with POAG and controls. A statistically significant difference was observed in IL-1ra, IL-2, IL-5, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17A, Eotaxin, FGF basic, G-CSF, GM-CSF, IFN-γ, MIP-1α, PDGF-bb, MIP-1β, RANTES, TNF-α, and VEGF. PCG patients have a cytokine profile in aqueous humor different from POAG patients and patients without glaucoma, characterized by lower concentrations of multiple cytokines.

Critical role of transcriptome-wide m6A methylation in the aqueous humor of patients with pseudoexfoliation glaucoma

N6-methyladenosine (m6A) modification is one of the most common types of methylation modifications in eukaryotic mRNA. However, its role in the pathogenesis of pseudoexfoliation glaucoma (PXG) has not yet been reported. To enhance understanding in this regard, we assessed the m6A methylome in the aqueous humor of patients with PXG. MeRIP-Seq and RNA-Seq analyses were performed to compare the m6A methylomes and gene expression profiles of the aqueous humor of patients with PXG with those of patients with age-related cataract (ARC). Colorimetric m6A quantification was performed to detect global m6A levels. Quantitative reverse transcription PCR confirmed the expression of m6A-related enzymes and mRNAs in both groups. Results showed significantly higher aqueous humor m6A levels in the PXG group than in the ARC group. Five m6A-related enzymes, including METTL3, YTHDC2, HNRNPA2B1, HNRNPC, and LRPPRC, were significantly up-regulated in PXG specimens. We also observed 9728 m6A-modified peaks related to 6126 gene transcripts in the PXG group, with more than 250 genes containing one m6A peak (hypomethylated or hypermethylated). The distribution of the m6A peaks was enriched in coding sequences and 3′-untranslated regions for both groups. GGAC motif structures were also significantly enriched. Bioinformatics analysis further revealed that m6A plays a critical role in extracellular matrix formation and histone deacetylation. Additionally, MMP14, ADAMTSL1, FN1, and HDAC1 showed significant changes in m6A methylation and mRNA expression in the PXG group. Therefore, m6A methylation may regulate extracellular matrix composition in PXG and METTL3 may be a pivotal regulator of this process. In the future, it would be necessary to investigate MMP14, ADAMTSL1, FN1, and HDAC1, which are potential target genes.

VEGF levels in the aqueous humor of patients with primary open angle glaucoma: A systematic review and a meta-analysis.

To compare the VEGF levels in the aqueous humor of patients with Primary Open Angle Glaucoma (POAG) and non-glaucomatous eyes and reveal any potential statistically significant correlations. We searched PubMed, from inception to December 31, 2021. Key search terms included VEGF and Glaucoma. All relevant studies that evaluated the VEGF levels in patients with POAG and in the control group were included in this systematic review. The Preferred Reporting Items for Systematic Reviews and Meta-Analysis of Diagnostic Test Accuracy Studies (PRISMA-DTA) guidelines were followed. Data were extracted independently by 2 authors. Heterogeneity was statistically quantified by Q, H, and I2 statistics, and a meta-analysis was performed using the random-effects model. Seven cross-sectional studies were included in the meta-analysis. 144 eyes were enrolled in the POAG group and 162 eyes in the control group. The random effect model showed no statistically significant difference between the two groups (SMD =0.284, 95% CI = -0.173 to 0.741; P = 0.223), but we noticed a trend towards elevated VEGF levels in the aqueous humor of POAG patients. Significant heterogeneity was detected (I2 = 74.1%, P = 0.001). This systematic review and meta-analysis indicates a trend towards elevated VEGF-A levels in the aqueous humor of patients with POAG and suggests a potential neuroprotective role of VEGF in patients with POAG. Future studies are required to evaluate the exact role of VEGF in POAG.

Expression of miR-210-3p in the aqueous humor of patients with age-related cataracts and its effect on human lens epithelial cell injury induced by hydrogen peroxide

Purpose: The regulatory effect of microRNA on diseases has been confirmed. This study aimed to evaluate the expression of microRNA-210-3p in age-related cataracts and assess the effect of abnormal miR-210-3p expressions on H2O2-induced SAR01/04 cells. Methods: Reverse-transcription quantitative polymerase chain reaction method was performed to assess the levels of miR-210-3p in aqueous humor samples. Receiver operating characteristic analysis was employed to assess the discrimination ability of miR-210-3p between patients with age-related cataracts and healthy people, and Pearson correlation analysis was used to identify the correlation between miR-210-3p and oxidative stress indices such as superoxide dismutase, glutathione peroxidase, malonaldehyde. Cell counting kit-8 assay and Transwell assay were used to estimate the biological function of H2O2-induced age-related cataract cell model. The levels of oxidative stress indices such as superoxide dismutase, glutathione peroxidase, and malonaldehyde were measured to evaluate the degree of oxidative stress damage in the age-related cataract cell model. The relationship between miR-210-3p and its target gene was verified by luciferase reporter gene analysis. Results: The miR-210-3p expression was elevated in the aqueous humor of patients with age-related cataracts. A high miR-210-3p expression showed a high diagnostic value for age-related cataracts and was significantly associated with the level of oxidative stress markers in patients with age-related cataracts. The inhibition of miR-210-3p can reverse oxidative stress stimulation and adverse effects on H2O2-induced cell function. Conclusions: The results suggested that miR-210-3p could promote cell viability, cell migration, and oxidative stress by targeting autophagy-related gene 7 in in vitro age-related cataract cell model.

The role of oxidative damage in cataract etiopathogenesis.

Cataract usually occurs due to age and diabetes, but the mechanisms of cataract formation have not yet been fully elucidated. In this study, the relationship between cataract and oxidative stress was evaluated by examining the aqueous humor reflecting lens metabolism. In this study, the effect of oxidative stress on the etiopathogenesis of cataract was investigated through the total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI), and arylesterase (ARE) levels in aqueous humor samples of patients with cataract. A prospective cohort study. This study was conducted on patients who were scheduled for cataract surgery between June 2020 and March 2021. The patients were divided into four groups according to their cataract density as grades 1, 2, 3, and 4. TOS, TAS, and ARE levels of aqueous humor samples were measured spectrophotometrically, and comparisons were made between groups. A total of 100 eyes of 100 patients were included in this study. TAS levels were found significantly higher in the grade 2 group compared with the grade 4 group (p = 0.006). In addition, a significant negative correlation was present between cataract grade and TAS level (r = -0.237; p = 0.018). There was no significant difference between diabetic and nondiabetic patients in terms of TAS, TOS, OSI, and ARE levels. The aqueous humor of patients with a high degree of cataract is characterized by low antioxidant capacity. Decreased antioxidant capacity has a role in cataract formation and progression.

Trimethylamine-N-Oxide Promotes High-Glucose-Induced Dysfunction and NLRP3 Inflammasome Activation in Retinal Microvascular Endothelial Cells.

Along with blood glucose levels, diabetic retinopathy (DR) development also involves endogenous risk factors, such as trimethylamine-N-oxide (TMAO), a product of intestinal flora metabolic disorder, which exacerbates diabetic microvascular complications. However, the effect of TMAO on retinal cells under high-glucose conditions remains unclear. Therefore, this study examined the effects of TMAO on high-glucose-induced retinal dysfunction in the context of NLRP3 inflammasome activation, which is involved in DR. TMAO was assessed in the serum and aqueous humor of patients using ELISA. Human retinal microvascular endothelial cells (HRMECs) were treated for 72 h as follows: NG (normal glucose, D-glucose 5.5 mM), NG + TMAO (5 μM), HG (high glucose, D-glucose 30 mM), and HG + TMAO (5 μM). The CCK8 assay was then used to assess cell proliferation; wound healing, cell migration, and tube formation assays were used to verify changes in cell phenotype. ZO-1 expression was determined using immunofluorescence and western blotting. Reactive oxygen species (ROS) formation was assessed using DCFH-DA. NLRP3 inflammasome complex activation was determined using a western blot. The serum and aqueous humor from patients with PDR contained higher levels of TMAO compared to patients with nontype 2 diabetes (Control), non-DR (NDR), and non-PDR (NPDR). TMAO showed significant acceleration of high-glucose-induced cell proliferation, wound healing, cell migration, and tube formation. ZO-1 expression decreased remarkably with the combined action of TMAO and a high glucose compared to either treatment alone. TMAO also promoted high-glucose-activated NLRP3 inflammasome complex. The combination of TMAO and high-glucose results in increased levels of ROS and NLRP3 inflammasome complex activation in HRMECs, leading to exacerbated retinal dysfunction and barrier failure. Thus, TMAO can accelerate PDR occurrence and development, thus indicating the need for early fundus monitoring in diabetic patients with intestinal flora disorders.

Antioxidant and free radicals species in the aqueous humor of patients with age-related cataract

Introduction/Objective. Age-related cataract is a significant cause of visual impairment worldwide. Oxidative damage and the effects of free radical species are considered important in the etiopathogenesis of cataracts. The aim of this study was to evaluate the antioxidative capacity and oxidative stress in the aqueous humor (AH) according to age and cataracts maturity. Methods. Clinical and biochemical researches were carried out in 55 patients with age-related cataract. According to the cataract maturity, patients were classified into incipient (cortical ? group C, 18 patients; nuclear ? group N, 20 patients; mature ? group M, 17 patients). In order to evaluate the impact of age patients within each group were divided into Group I (65?69 years) and Group II (70 ? years). The antioxidant activity of AH was measured by the reduction power method and the activity of glutathione peroxidase (GPx) spectrophotometrically. Changes in the concentrations of hydroxyl and ascorbyl radicals were detected by electron spin resonance spectroscopy. Results. Both reduction power and GPx activity were significantly (p < 0.001) reduced in group N compared to group C and in group M compared to group N. Concentrations of hydroxyl (29.45 ? 1.01% in group C, 38.12 ? 1.29% in group N, and 74.14 ? 2.52% in group M) and ascorbyl radicals (26.12 ? 0.89% in group C, 41.15 ? 1.39% in group N, and 83.56 ? 2.84% in group M) increased significantly (p < 0.05) with progression of age-related cataract. Significant negative correlation (r = -0.759, p < 0.05) was determined between concentrations of HO? and content of GPx. Conclusion. Our research proved that the level of oxidative stress in the AH is significantly affected during aging and cataract progression, The obtained data support the hypothesis that during aging, depending on the maturity of the cataract, the antioxidant capacity in the AH decreases due to an increase in the concentration of reactive HO?.

Correlation of photoreceptor damage with anti-retina antibodies level in aqueous humor in macular edema patients.

This study aimed to investigate the correlation between the severity of photoreceptor damage and the level of anti-retina antibodies (ARAs) in aqueous humor, including recoverin, CA II and enolase-α IgG antibody of macular edema patients. Aqueous humor samples were collected from macular edema patients and from cataract patients. Patients were divided into three groups according to the severity of discontinuity of ellipsoid zone (EZ) shown on optical coherence tomography (OCT) imaging: cataract patients with intact EZ, macular edema patients with mild EZ damage, and macular edema patients with severe EZ damage. The level of ARAs was determined with enzyme-linked immunosorbent assay (ELISA). The correlation between the level of ARAs and the degree of photoreceptor damage was analyzed. The level of ARAs of the intact EZ group was significantly lower than that in the severely damaged group (P < 0.05). The level of recoverin IgG of the intact EZ group was significantly lower than mildly damaged group (P = 0.030). In a subgroup analysis, the level of recoverin IgG of DME patients was correlated with their central retinal thickness (CRT) (r = 0.462, P = 0.035). The level of ARAs in aqueous humor of patients with DME and RVO-ME was correlated with the degree of photoreceptor damage.

VEGF levels in the aqueous humour of patients with primary open angle glaucoma: A systematic review and a meta‐analysis

AbstractPurpose: To compare the VEGF levels in the aqueous humour of patients with Primary Open Angle Glaucoma (POAG) and non‐glaucomatous eyes and reveal any potential statistically significant correlations.Methods: We searched PubMed, from inception to December 31, 2021. Key search terms included VEGF and Glaucoma. All relevant studies that evaluated the VEGF levels in patients with POAG and in the control group were included in this systematic review. The Preferred Reporting Items for Systematic Reviews and Meta‐Analysis of Diagnostic Test Accuracy Studies (PRISMA‐DTA) guidelines were followed. Data were extracted independently by 2 authors. Heterogeneity was statistically quantified by Q, H, and I2 statistics, and a meta‐analysis was performed using the random‐effects model.Results: Seven cross‐sectional studies were included in the meta‐analysis. 144 eyes were enrolled in the POAG group and 162 eyes in the control group. The random effect model showed no statistically significant difference between the two groups (SMD = 0.284, 95% CI = −0.173 to 0.741; p = 0.223), but we noticed a trend towards elevated VEGF levels in the aqueous humour of POAG patients. Significant heterogeneity was detected (I2 = 74.1%, p = 0.001).Conclusions: This systematic review and meta‐analysis indicates a trend towards elevated VEGF‐A levels in the aqueous humour of patients with POAG and suggests a potential neuroprotective role of VEGF in patients with POAG. Future studies are required to evaluate the exact role of VEGF in POAG.

Correlation between inflammatory profile in aqueous humour in patients who underwent cataract surgery using two techniques and cataract degree

AbstractPurpose: Cataract surgery is one of the most usual type of intervention worldwide. Thanks to the improvement of technology and investigation this kind of surgery has reached a very high success rate. Studies have compared laser and conventional techniques in terms of inflammation, but there are also other variables which can influence the final result, such as cataract degree, age or diabetes. The purpose of this study is to establish the influence of cataract degree in the inflammatory profile of the aqueous humour.Methods: The parameters determined in aqueous humour to compare both groups were interleukins (IL) 6 and 8 and cytokine tumour necrosis factor α (TNFα). Samples of aqueous humour were obtained during cataract surgery, before the lens was extracted and were analysed through Enzyme‐Linked Immunosorbent Assay (ELISA). Two study groups were established: femtosecond group (75 samples of aqueous humour from 75 patients) and manual group (54 samples of aqueous humour from 54 patients). Once this classification was made, the non‐parametric statistics of the median, Kolmogorov–Smirnov and the Mann–Whitney test were calculated to verify whether the degree of cataract influenced the inflammatory profile of aqueous humour.Results: No significative differences were noticed between the two types of cataract degrees, early and advanced and their inflammatory reaction to surgery, although there is a slight tendency in IL6 and IL8 to increase their concentrations in advanced cataracts. On the other hand, the opposite happens with the concentration of TNFα that diminishes in advanced cataracts.Conclusions: The inflammatory profile of aqueous humour can be affected by the type of surgery performed (femtosecond or manual surgery) but also by other variables such as age, diabetes or cataract degree. Regarding the last one, cataract itself can modulate ocular inflammation as there was a tendency in IL6 and IL8 to increase in advanced cataracts. Besides that, the decrease in TNFα aqueous humour concentrations in advanced cataracts may be due to a physiological attempt to restore basal cytokine levels.

High Levels of Heat Shock Protein 47 in the Aqueous Humor of Patients with Acute Primary Angle Closure

Introduction: Acute primary angle closure (APAC) is often characterized by acute elevation of intraocular pressure accompanied by severe ocular and systemic symptoms. Excessive collagen accumulation, which can be caused by upregulated heat shock protein 47 (HSP47) expression, can produce scarring in rat conjunctival blebs. Meanwhile, the presence of HSP47 in human aqueous humor and its levels are yet to be determined. Methods: We examined 32 consecutive patients with APAC and 16 age-matched participants without APAC scheduled for cataract surgery who were enrolled as a control group. Aqueous humor samples were collected from all subjects at the time of surgery and compared between the subjects with and without APAC. Results: The levels of HSP47 in the aqueous humor of patients with APAC (1,210.4 ± 450.2 pg/mL) were found to be significantly increased (p = 0.001) compared with those in the control group (863.4 ± 240.0 pg/mL). Notably, the levels of HSP47 negatively correlated with the age of patients with APAC (p = 0.023). Conclusion: HSP47 was upregulated in the aqueous humor of patients with APAC and may play a role in scarring after trabeculectomy for APAC.

Factors Associated with Elevated Tumor Necrosis Factor-α in Aqueous Humor of Patients with Open-Angle Glaucoma.

Tumor necrosis factor-alpha (TNF-α) is an important modulator of neuroinflammation, secreted from activated glial cells in response to intraocular stress. The purpose of this study was to investigate the clinical factors associated with elevated TNF-α and its level in aqueous humor of patients with open-angle glaucoma (OAG). Aqueous humor was collected from 73 OAG eyes, and TNF-α level was analyzed using the singleplex bead immunoassay method. Patients were divided into TNF-α-positive and TNF-α-negative groups according to the TNF-α level of 10 pg/mL, and baseline clinical characteristics were compared. The TNF-α-positive group showed higher baseline IOP, greater IOP fluctuation, and higher systolic blood pressure than the TNF-α-negative group (p = 0.007, p < 0.001, and p = 0.009, respectively). In the multivariate logistic regression analysis, IOP fluctuation (p = 0.037) and systolic blood pressure (p = 0.016) were all independently associated with positive TNF-α level. In normal-tension glaucoma (NTG) patients, presence of central scotoma (p = 0.029) was significantly associated with positive TNF-α level. In conclusion, positive TNF-α level in OAG patients was associated with greater IOP fluctuation and higher systolic blood pressure. In NTG patients, positive TNF-α level was associated with the presence of central scotoma. IOP factors and vascular factors, including blood pressure and presence of central scotoma, may indicate glaucoma pathogenesis related to TNF-α elevation in OAG patients.

Analysis of the Cytokine Expression in the Aqueous Humor of Individuals with BRVO-Associated Macular Edema.

Purpose This study aimed to determine the expression levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in the aqueous humor of patients with macular edema (ME) caused by branch retinal vein occlusion (BRVO), as well as to investigate the relationship between the cytokines as mentioned earlier and best-corrected visual acuity (BCVA), ME, and the degree of ME from the molecular level. Methods In a prospective observational study, fluorescein fundus angiography (FFA) and optical coherence tomography (OCT) were used to classify 58 patients with non-ischemic BRVO-ME into three groups according to the degree of ME: 14-mild, 17-moderate, and 27-severe. The specific concentration of IL-6, VEGF, ICAM-1, and VCAM-1 in the aqueous humor was detected using the BD CSCanto™ II Flow Cytometer (US). Spearman or Pearson correlation analysis was used to test the correlation between the levels of BCVA and severity of ME and the expression levels of IL-6, VEGF, ICAM-1, and VCAM-1 in the aqueous humor. Results According to the obtained data, BCVA did not correlate with the severity of ME, and these four cytokines expression levels in patients' aqueous humor (P > 0.05). Moreover, BCVA did not correlate with mild, moderate, or severe ME as well (P > 0.05). However, the levels of these four cytokines were correlated with the severity of the ME. These underlined cytokines were linked to the mild, moderate, and severe degrees of ME. VEGF was also significantly correlated (r > 0.8, P < 0.0001) with the severity of ME. Conclusions This study suggests that the severity of ME in BRVO-ME patients is significantly correlated with the expression levels of IL-6, VEGF, ICAM-1, and VCAM-1 in the aqueous humor. Lowering the level of disease-associated cytokines may potentially reduce the degree of ME. Therefore, an in-depth study of the levels and the relationship may provide some evidence for the pathogenesis, treatment, and prevention of BRVO-ME.

The Role of Oxidative Status in the Pathogenesis of Primary Open-Angle Glaucoma, Pseudoexfolyation Syndrome and Glaucoma.

To investigate the role of oxidative stress and antioxidant system in the etiopathogenesis of pseudoexfoliation and progression of pseudoexfoliation syndrome to glaucoma. A total of 20 patients with pseudoexfoliation syndrome, 20 with pseudoexfoliation glaucoma, 20 with primary open-angle glaucoma, and 20 without pseudoexfoliation and glaucoma as a control group, who underwent cataract or glaucoma surgery between December 2020 and March 2021 in the Health Sciences University Beyoğlu Eye Training and Research Hospital, 80 patients were included in the study. Best corrected visual acuity (BCVA), intraocular pressure with applanation tonometry, detailed anterior and posterior segment examinations with biomicroscopy were performed in all patients included in the study. Approximately 0.1 cc of anterior chamber fluid was taken from all patients at the beginning of surgery. Total Oxidant Status (TOS) and Total Antioxidant Status (TAS) levels were measured by keeping the samples taken in the deep freezer of the cornea bank at -80 degrees, transferring them with cold chain transport rules, and examining them with automatic measurement method in the laboratories of the Medical Biochemistry Department of the University of Health Sciences. Oxidative Stress Index (OSI) value was calculated to measure the degree of oxidative stress. TAS averages of the control and POAG groups were found to be statistically significantly higher than the PES and PEG groups (p = 0.0001, p = 0.0001). No significant difference was observed between the other groups (p > 0.05). The TOS averages of the control and POAG groups were found to be statistically significantly lower than the PES and PEG groups (p = 0.003, p = 0.0001; p = 0.01, p = 0.001), no statistically significant difference was observed between the other groups (p > 0.05). The OSI mean of the control and POAG groups was found to be statistically significantly lower than the PES and PEG groups (p = 0.001, p = 0.0001; p = 0.002, p = 0.0001), no statistically significant difference was observed between the other groups (p > 0.05). Increased TOS and OSI and decreased TAS levels in the aqueous humour of patients with PES and PEG suggest that increased oxidative stress and decreased antioxidative defense system play a role in the etiopathogenesis of the disease.

Clinical Metagenomic Next-Generation Sequencing for Diagnosis of Secondary Glaucoma in Patients With Cytomegalovirus-Induced Corneal Endotheliitis.

Glaucoma is the second leading cause of blindness globally. Growing scientific evidence indicated that inflammation of the trabecular meshwork induced by corneal endotheliitis could lead to secondary glaucoma. Cytomegalovirus (CMV) has been identified as the most common herpes virus in corneal endotheliitis patients. Early detection is critical in preventing endothelial cell loss, and patient management should vary based on different pathological factors. However, routine culture and real-time polymerase chain reaction (qPCR) have difficult in distinguishing whether CMV, Varicella Zoster Virus (VZV) or Herpes Simplex Virus (HSV) causes endothiliitis. This may result in inappropriate treatment, which may prolong or aggravate the status of disease. We compared the sensitivity and specificity of qPCR and Metagenomic Next-Generation Sequencing (mNGS) in the aqueous humor of patients with suspected CMV endotheliitis in this study. Our results showed that four out of 11 (36.4%) of our patients were positive for CMV by qPCR, whereas mNGS had a 100% detection rate of CMV. Our findings implied that mNGS could be a useful diagnostic tool for CMV-induced endotheliitis.

Silencing of circular RNA‑ZYG11B exerts a neuroprotective effect against retinal neurodegeneration.

Ischemic retinal diseases are the major cause of vision impairment worldwide. Currently, there are no available treatments for ischemia-induced retinal neurodegeneration. Circular RNAs (circRNAs) have emerged as important regulators of several biological processes and human diseases. The present study investigated the role of circRNA-ZYG11B (circ-ZYG11B; hsa_circ_0003739) in retinal neurodegeneration. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) demonstrated that circZYG11B expression was markedly increased during retinal neurodegeneration in vivo and in vitro. Cell Counting Kit-8, TUNEL and caspase-3 activity assays revealed that silencing of circZYG11B was able to protect against oxidative stress- or hypoxic stress-induced retinal ganglion cell (RGC) injury. Furthermore, immunofluorescence staining and hematoxylin and eosin staining revealed that silencing of circZYG11B alleviated ischemia/reperfusion-induced retinal neurodegeneration, as indicated by reduced RGC injury and decreased retinal reactive gliosis. In addition, luciferase reporter, biotin-coupled miRNA capture and RNA immunoprecipitation assays revealed that circZYG11B could regulate RGC function through circZYG11B/microRNA-620/PTEN signaling. Clinically, RT-qPCR assays demonstrated that circZYG11B expression was markedly increased in the aqueous humor of patients with glaucoma. In conclusion, circZYG11B may be considered a promising target for the diagnosis and treatment of retinal ischemic diseases.

Downregulation of angiogenic factors in aqueous humor associated with less intraoperative bleeding in PDR patients with NVG receiving conbercept: a randomized controlled trial

BackgroundTo analyze the level changes of 28 cytokines in aqueous humor of patients with proliferative diabetic retinopathy (PDR) coexisting neovascular glaucoma (NVG) after intravitreal injection of conbercept (IVC), and to investigate whether these cytokines are associated with intraoperative bleeding (IOB).MethodsTotally 34 eyes with NVG secondary to PDR were enrolled. Patients were randomized into two groups, and all of them underwent 25-gauge pars plana vitrectomy (PPV) combined with trabeculectomy. Group I, 18 eyes received IVC 3 days before PPV, and 100 µL aqueous humor was collected at the time of IVC pretreatment and 3 days later at the beginning of PPV respectively. Group II, 16 eyes received IVC after PPV, and 100 µL aqueous humor was collected only at the beginning of PPV. Aqueous humor from 19 eyes with age-matched cataract patients served as controls. Luminex bead-based multiplex array was used to measure the levels of 28 cytokines in aqueous humor. The baseline cytokine levels were compared among the three groups. All NVG patients were divided into IOB and non-bleeding (INB) groups. The cytokine levels of aqueous humor at the beginning of PPV were compared between group I and II, also between IOB and INB groups. IOB in NVG patients was graded according to vitreous bleeding amount. The correlation between cytokine levels and the grades of IOB were analyzed.ResultsCompared with controls, the baseline levels of 18 cytokines associated with inflammation and angiogenesis showed significantly increased in group I and group II (all, P < 0.0167). The IOB rate as well as the levels of IL-4, IL-22, Ang-2, PLGF and VEGF-A in group I were significantly lower than in group II (all, P < 0.05). The levels of IL-4, IL-22, Ang-2, PLGF and VEGF-A were significantly lower in INB group than in IOB group (all, P < 0.05). The levels of IL-4, Ang-2, PLGF and VEGF-A were positively correlated with the grades of IOB in NVG patients (all, rs > 0.4, P < 0.05).ConclusionsIVC 3 days before PPV combined with trabeculectomy reduces IOB in NVG patients, in which the downregulation of IL-4, Ang-2, PLGF and VEGF-A after IVC may be an underlying mechanism.Trial registrationChiCTR2100048118, retrospectively registered on 2 July 2021.

Status of higher TGF-\u03b21 and TGF-\u03b22 levels in the aqueous humour of patients with diabetes and cataracts

BackgroundTransforming growth factor (TGF) is a cytokine that acts on the proliferation, migration, differentiation, and apoptosis of cells and the accumulation of extracellular matrix components. Very few studies have precisely evaluated the concentration of TGF-β in the aqueous humour (AH) of diabetic and cataract (DMC) eyes due to the low expression of proteins in the AH or other reasons. The concentrations of TGF-β1, -β2, and -β3 in the AH of the DMC group were compared with those of the age-related cataract (ARC) group.MethodsWe collected AH and lens epithelium samples from 33 DMC patients and 36 ARC patients. Luminex liquid suspension chip detection was applied to detect the concentration of TGF-β1, -β2, and -β3 in the AH samples. The expression of TGFB1/2/3 in lens epithelium samples was determined by quantitative real-time polymerase chain reaction (qRT-PCR).ResultsThe concentrations of TGF-β1 and TGF-β2 in AH samples of DMC eyes were higher than those of ARC eyes. The differences in TGF-β1 and TGF-β2 between the two groups were statistically significant (P value = 0.001 for TGF-β1, P value = 0.023 for TGF-β2). The difference of the correlation between TGF-β1 and glycosylated haemoglobin was significant (P value = 0.011, and Pearson correlation coefficient = 0.306). The difference of the correlation between TGF-β2 and glycosylated haemoglobin was significant (P value = 0.026, and Pearson correlation coefficient = 0.269). The mRNA expression levels of TGFB1 and TGFB2 were upregulated in DMC epithelium samples compared with ARC epithelium samples. The differences in TGFB1 and TGFB2 between the two groups were statistically significant (P value for TGFB1 = 0.041, P value for TGFB2 = 0.021).ConclusionsThe concentrations of TGF-β1 and TGF-β2 in AH samples were significantly higher in DMC eyes than in ARC eyes. The higher the glycosylated haemoglobin was, the higher the concentrations of TGF-β1 and -β2 were. The mRNA expression of TGFB1 and TGFB2 was significantly upregulated in DMC epithelial samples compared with ARC epithelial samples, suggesting the proinflammatory status of the anterior chamber of DMC eyes.

Levels of IL-8 and IL-12p70 in the aqueous humor of patients with primary acute angle-closure glaucoma and its clinical significance

Levels of IL-8 and IL-12p70 in the aqueous humor of patients with primary acute angle-closure glaucoma and its clinical significance