Increased Risk of Ocular Hypertension in Patients With Cushing's Disease.

PRCIS: An increased risk of ocular hypertension was seen in Cushing's disease. INTRODUCTION: Systemic steroid use is a significant risk factor for increased intraocular pressure (IOP). The incidence of ocular hypertension may rise to 30%-40% of the general population due to topical or systemic glucocorticoid usage. However, the incidence of ocular hypertension in endogenous hypercortisolemia, as well as the ophthalmological outcomes after endocrine remission due to surgical resection, remain unknown. MATERIALS AND METHODS: The IOP, visual field, and peripapillary retinal nerve fiber layer thickness were documented in all patients with Cushing's disease (CD) admitted to a tertiary pituitary center for surgery from January to July 2019. Patients with acromegaly and patients with nonfunctioning pituitary adenoma (NFPA) during the same study period served as controls. We calculated the odds ratio (OR), identified the risk factors of developing ocular hypertension, and presented postoperative trends of the IOP. RESULTS: A total of 52 patients (38.4±12.4 y old) with CD were included. The IOP was higher in patients with CD (left 19.4±5.4 mm Hg and right 20.0±7.1 mm Hg) than in patients with acromegaly (left 17.5±2.3 mm Hg and right 18.6±7.0 mm Hg, P =0.033) and patients with NFPA (left 17.8±2.6 mm Hg and right 17.4±2.4 mm Hg, P =0.005). A total of 21 eyes (20.2%) in patients with CD were diagnosed with ocular hypertension compared with 4 eyes (4.7%) in the acromegaly group and 4 eyes (4.5%) in the NFPA group. The OR of developing ocular hypertension in patients with CD was 5.1 [95% confidence interval (CI), 1.3-25.1, P =0.029] and 6.6 (95% CI, 1.8-30.3, P =0.007) when compared with the 2 control groups. Among patients with CD, those with a higher urine-free cortisol were more likely to develop ocular hypertension (OR=19.4, 95% CI, 1.7-72.6). The IOP decreased at 1 month after surgery in patients with CD, and the change was sustained for 3 months after surgery. CONCLUSIONS: An increased risk of ocular hypertension was seen in CD and suggests that endogenous hypercortisolemia should be considered as part of the glaucoma assessment. This result warrants the discretion of both ophthalmologists and neuroendocrinologists.

Machine Learning Prediction of Visual Outcome after Surgical Decompression of Sellar Region Tumors.

INTRODUCTION: This study aims to develop a machine learning-based model integrating clinical and ophthalmic features to predict visual outcomes after transsphenoidal resection of sellar region tumors. METHODS: Adult patients with optic chiasm compression by a sellar region tumor were examined to develop a model, and an independent retrospective cohort and a prospective cohort were used to validate our model. Predictors included demographic information, and ophthalmic and laboratory test results. We defined "recovery" as more than 5% for a p-value in mean deviation compared with the general population in the follow-up. Seven machine learning classifiers were employed, and the best-performing algorithm was selected. A decision curve analysis was used to assess the clinical usefulness of our model by estimating net benefit. We developed a nomogram based on essential features ranked by the SHAP score. RESULTS: We included 159 patients (57.2% male), and the mean age was 42.3 years old. Among them, 96 patients were craniopharyngiomas and 63 patients were pituitary adenomas. Larger tumors (3.3 cm vs. 2.8 cm in tumor height) and craniopharyngiomas (73.6%) were associated with a worse prognosis (p < 0.001). Eyes with better outcomes were those with better visual field and thicker ganglion cell layer before operation. The ensemble model yielded the highest AUC of 0.911 [95% CI, 0.885-0.938], and the corresponding accuracy was 84.3%, with 0.863 in sensitivity and 0.820 in specificity. The model yielded AUCs of 0.861 and 0.843 in the two validation cohorts. Our model provided greater net benefit than the competing extremes of intervening in all or no patients in the decision curve analysis. A model explanation using SHAP score demonstrated that visual field, ganglion cell layer, tumor height, total thyroxine, and diagnosis were the most important features in predicting visual outcome. CONCLUSION: SHAP score can be a valuable resource for healthcare professionals in identifying patients with a higher risk of persistent visual deficit. The large-scale and prospective application of the proposed model would strengthen its clinical utility and universal applicability in practice.

5-Aza-2'-deoxycytidine induces human Tenon's capsule fibroblasts differentiation and fibrosis by up-regulating TGF-ß type I receptor.

The principle reason of high failure rate of glaucoma filtration surgery is the loss of filtration function caused by postoperative scar formation. We investigated the effects of 5-aza-2'-deoxycytidine (5-Aza-dc), a DNA methyltransferases inhibitor, on human Tenon's capsule fibroblasts (HTFs) differentiation and fibrosis and its mechanism of action, especially in relation to transforming growth factor (TGF)-ß1 signaling. TGF-ß1 was used to induce differentiation of cultured HTFs. 5-Aza-dc suppressed DNA methyltransferases (DNMTs) activity 6 h after treatment with a course corresponding to that of TGF-ß1-induced reduction of DNMT activity without affecting cell viability as measured by Cell Counting Kit-8 assay. 5-Aza-dc also reduced DNMT1 and DNMT3a protein expression from 24 to 48 h. HTFs migration evaluated by scratch-wound assay were significantly increased 24 h after 5-Aza-dc treatment, a time course similar to that of TGF-ß1. Treatment with 5-Aza-dc significantly increased the mRNA and protein levels of α-smooth muscle actin (α-SMA), collagen-1A1 (Col1A1), fibronectin (FN) and TGF-ß type I receptor (TGFßRI). Furthermore, the effects of 5-Aza-dc on DNMT activity suppression, cell migration, and fibrosis were all reversed by a TGFßRI inhibitor- SB-431542. Meanwhile, knockdown of DNMT1 upregulated TGFßRI expression and had the same fibrosis-inducing effect in HTFs, which was also inhibited by SB-431542. Thus, the results indicate that DNA hypomethylation induces HTFs differentiation and fibrosis through up-regulation of TGFßRI. DNA methylation status plays an important role in subconjunctival wound healing.

Overexpression of ALK5 Induces Human Tenon's Capsule Fibroblasts Transdifferentiation and Fibrosis In Vitro.

PURPOSE: To investigate the involvement of activin receptor-like kinase 5 (ALK5) in human Tenon's capsule fibroblasts (HTFs) transdifferentiation and fibrosis. METHODS: (1) Cultured HTFs were treated with transforming growth factor beta 1 (TGF-ß1) at different concentrations for different durations, mRNA expression of ALK5 and plasminogen activator inhibitor-1 (PAI-1) was measured by quantitative polymerase chain reaction (PCR) while protein expression of ALK5, α-smooth muscle actin (α-SMA), and extracellular matrix deposition including fibronectin (FN) and collagen I (Col1) was assessed by western blot. HTFs with or without TGF-ß1 were also treated with an ALK5 activity inhibitor, SB-431542, and fibrosis-related genes were assessed. (2) HTFs were transduced with ALK5 lentivirus (ALK5-OE group) or empty lentivirus (NC-OE) with or without the treatment of SB-431542. Protein expression of ALK5, α-SMA, FN, and Col1 was evaluated. (3) HTFs in the ALK5-OE group and NC-OE group were subjected to a scratch-wound assay and their migratory activities assessed. RESULTS: (1) TGF-ß1, in a concentration-dependent manner, upregulated ALK5 and PAI-1 expressions in the HTFs, which peaked between 24 and 36 h. These changes were associated with increases in protein levels of FN, Col1, and α-SMA. These TGF-ß1 effects were blocked by the ALK5 inhibitor SB-431542. (2) Similarly, overexpression of ALK5 by lentiviral vector significantly increased protein expression of α-SMA, FN, and Col1. Addition of TGF-ß1 to the ALK5-OE cells did not produce additional expression of any of the marker proteins. The upregulation of extracellular matrix and α-SMA can be reduced by SB-431542. (3) In ALK5-OE group, HTFs migration was significantly increased compared with normal control and TGF-ß1 could still promote ALK5-OE cells migration. CONCLUSIONS: Our findings suggest that ALK5 is an important mediator of HTFs fibrosis. ALK5 is a potential therapeutic target to suppress scar formation after filtration surgery.

Angiotensin II as a morphogenic cytokine stimulating fibrogenesis of human tenon's capsule fibroblasts.

PURPOSE: To examine the expression of Angiotensin II (Ang II) and its type I, and type II receptors (AT1R, AT2R) in rabbit Tenon's capsule fibroblasts after trabeculectomy, and to investigate the effects of Ang II on cultured human Tenon's capsule fibroblasts (HTFs) proliferation, migration, phenotype transition, and extracellular matrix (ECM) synthesis. METHODS: In the rabbit, expression of Ang II, AT1R, and AT2R in Tenon's capsule fibroblasts of eyes after trabeculectomy was evaluated by immunohistochemistry. Ang II levels in aqueous humor and plasma were assessed by ELISA. Cultured HTFs, obtained from patients undergoing cataract surgery, were treated with Ang II, TGF-ß1, or vehicle control. Cell proliferation and migration were evaluated by Cell Counting Kit-8 and Transwell assay, and wound scratch assay, respectively. Protein expressions of α-smooth muscle actin (α-SMA) and fibronectin (FN) were measured by Western blot and immunofluorescence. Messenger RNA expressions of α-SMA and FN were measured by real-time PCR. RESULTS: In the rabbit, the expression of Ang II and AT1R increased from 1 day after surgery while AT2R increased from 7 days. In cultured HTFs, Ang II promoted cell proliferation and migration significantly (P < 0.05). Interestingly, the effect of 10(-7) M Ang II was more prominent than higher concentrations (10(-5) M; P < 0.05). Ang II also markedly induced the expression of α-SMA and FN, suggesting a phenotypic transition to myofibroblasts. CONCLUSIONS: Our results show that trabeculectomy alter the levels of Ang II and its receptors in Tenon's capsule fibroblasts, and that Ang II increase HTFs proliferation, migration, and phenotype transition, suggesting that Ang II may play a role in wound healing after trabeculectomy.

Functional characteristics of connective tissue growth factor on human tenon's capsule fibroblast.

PURPOSE OF THE STUDY: To explore the characteristic of connective tissue growth factor (CTGF) on the phenotype transition, extracellular matrix (ECM) synthesis and proliferation of human Tenon's capsule fibroblasts (HTFs). MATERIALS AND METHODS: HTFs were obtained from patients during cataract surgery and induced by CTGF (1 to 100 µg/L). Western blot and immunofluorescence were performed to observe the expression of alpha smooth muscle actin (α-SM-actin) protein. The levels of mRNAs were quantified by real-time PCR. Col I and FN expression at both protein and RNA levels were tested after induction by CTGF and transforming growth factor ß (TGF-ß), respectively. Statistical significance was assumed if p < 0.05. RESULTS: CTGF upregulated the expression of α-SM-actin in cultured HTFs. Its maximum effect at protein level attained under the optimal concentration of 50 µg/L at the peak time of 48 hours, though still weaker than the effect of TGF-ß1 (10 µg/L, p < 0.05). The expression of Col I and FN at both protein and mRNA levels was elevated by the induction of CTGF (50 µg/L) (p < 0.01) and TGF-ß1 (10 µg/L) (p < 0.05), while CTGF (50 µg/L) showed a greater effect than the latter (p < 0.05). CTGF (1 to100 µg/L) increased the proliferation of HTFs significantly (p < 0.05). CONCLUSIONS: CTGF induced the phenotype transition of HTFs individually and significantly promoted their proliferation. Moreover, it promoted ECM synthesis, thus demonstrating its role as a crucial factor in fibrosis. Thus, CTGF could potentially be a safer and more efficient target than TGF-ß at suppressing scar formation after filtering surgery.

Protective effect of pioglitazone on retinal ischemia/reperfusion injury in rats.

PURPOSE: To investigate the protective effect of pioglitazone on the rat retina after ischemia/reperfusion (I/R) injury and to explore its possible mechanisms. METHODS: Retinal ischemia was induced by increasing the intraocular pressure to 110 mm Hg for 60 minutes, and pioglitazone was delivered 3 hours before the I/R. Retinal damage was quantified by measuring the thickness of the retina, the functional changes of visual evoked potential (VEP) and electroretinography (ERG), and the number of retinal ganglion cells (RGCs) at 7 days after I/R injury. Real-time PCR and Western blot analysis were performed to measure the glial fibrillary acidic protein (GFAP) expression. Retinal cell apoptosis was detected by TUNEL assay at 24 hours after reperfusion. Nuclear factor-κB (NF-κB), Bax, and Bcl-2 in the retina were determined by Western blot analysis. RESULTS: The I/R produced a degenerative effect primarily in the ganglion cell layer, inner plexiform layer, and inner nuclear layer. Pioglitazone maintained the retinal thickness, promoted the survival of RGCs, and attenuated the destruction of ERG and VEP caused by I/R. Pioglitazone pretreatment also suppressed NF-κB activation and altered GFAP overexpression. The number of TUNEL-labeled cells significantly decreased in the retinas pretreated with pioglitazone, and the Bax-Bcl-2 ratio was much lower in the retinas pretreated with pioglitazone than in the I/R group. CONCLUSIONS: Pioglitazone could inhibit activation of the glia cells, prevent cell apoptosis, and protect the retina from subsequent cellular damage caused by the retinal I/R. The possible mechanism might involve the NF-κB pathway.

SB-431542 inhibition of scar formation after filtration surgery and its potential mechanism.

PURPOSE: To explore the inhibitive effect of SB-431542 (an ALK5 inhibitor) on scar formation after glaucoma surgery and to identify the potential pharmacologic target(s). METHODS: Twenty-four New Zealand rabbits underwent filtration surgery on the right eye and were divided into a control group and three experimental groups (n=6). Human Tenon's fibroblast monolayer was scraped to generate a single gap, and then the control medium with SB-431542 only or containing 10 microg/L TGF-beta1 and SB-431542 (1-20 microM) was added. The cells were pretreated with SB-431542 or in control medium for 30 minutes before induction with 10 microg/L TGF-beta1 or 1 microg/L TGF-beta2. The expression of alpha-SM-actin, CTGF, and Col I, as well as changes in the Smad, ERK, P38, and AKT signaling pathways were detected. RESULTS: In comparison with the control rabbits, the IOPs in the experimental groups remained at lower levels until day 25 (P<0.05) after the surgery. Histologic profiles showed that there was only a mild deposition of collagen in the subconjunctival space in the experimental groups. The cell growth and migration were inhibited effectively by SB-431542, regardless of whether TGF-beta was present in the culture system. SB-431542 abrogated TGF-beta-induced upregulation of alpha-SM-actin, CTGF, and Col I. It effectively inhibited the phosphorylation of Smad2 stimulated by TGF-beta but not that of the components of the MAPK pathways. CONCLUSIONS: SB-431542 inhibits scar formation after glaucoma filtration surgery. The mechanism may be that SB-431542 interferes in the phosphorylation of Smad2, thus abrogating TGF-beta-induced fibroblast transdifferentiation and then decreasing Col I synthesis.

[Clinical and pathological analysis of 2734 cases of eyelid neoplasms].

OBJECTIVE: To study the histopathological classification and changes of eyelid neoplasms. METHOD: In this retrospective case series, the pathological specimens of 2734 cases with eyelid neoplasms examined between 1993-2005 were claimed and analyzed. RESULTS: There were 1248 eyelid tumors (45.65%) in 2734 cases with eyelid neoplasms, including 875 benign neoplasms (71.11%) and 960 malignant cases (29.89%). The three leading malignant eyelid tumors were basal cell carcinoma, meibomian gland carcinoma and squamous cell carcinoma The mean ages of the occurrence of these three tumors were 64.16, 63.30 and 60.38 years old, respectively. CONCLUSION: Pathologic classification of eyelid neoplasms is helpful for the pathological diagnosis and provides information for the diagnosis and treatment of these diseases.