Predicting visual outcomes in keratoprosthesis surgery with critical flicker fusion frequency, B-scan, visual electrophysiology and endoscopy.

PURPOSE: This study evaluates the predictive power of the critical flicker fusion frequency (CFF) test for visual outcomes in keratoprosthesis (KPro) candidates, comparing its accuracy with B-scan ultrasound, flash visual evoked potentials (fVEP) and endoscopy. METHODS: The study included 42 patients (42 eyes) scheduled for KPro surgery with a median follow-up period of 6 months. The receiver operating characteristic curve identified the cut-off threshold for CFF in the model development study (17 eyes). All patients in the comparison study (25 eyes) underwent preoperative assessments including trichromatic CFF (red, green and yellow), B-scan ultrasound, fVEP and perioperative endoscopy. Results were classified as either favourable or unfavourable predictors of visual outcomes based on predefined criteria. Sensitivity and specificity of each assessment were calculated based on postoperative best-corrected visual acuity (BCVA)≥20/200. The Bland-Altman test assessed the consistency between CFF-predicted BCVA and actual BCVA. RESULTS: Among the trichromatic CFF tests, the yellow-CFF (yCFF) exhibited the highest area under the curve value of 0.97 and a cut-off threshold at 10 Hz for predicting postoperative BCVA≥20/200 (p<0.05). yCFF achieved 90% sensitivity and 80% specificity in predicting satisfactory postoperative outcomes. Endoscopy had 80% sensitivity and 80% specificity, B-scan showed 70% sensitivity and 60% specificity, and fVEP had 75% sensitivity and 40% specificity. yCFF showed a mean bias of 0.091 logarithm of the minimum angle of resolution (logMAR) in postoperative prediction. CONCLUSIONS: The CFF test provides robust visual function evaluation in KPro candidates. It demonstrates superior predictive accuracy for visual prognosis compared with routine ophthalmologic examinations, such as B-scan ultrasonography, fVEP and endoscopy.

Adaptor protein 14-3-3zeta promotes corneal wound healing via regulating cell homeostasis, a potential novel therapy for corneal injury.

Severe corneal injury can lead to blindness even after prompt treatment. 14-3-3zeta, a member of an adaptor protein family, contributes to tissue repair by enhancing cellular viability and inhibiting fibrosis and inflammation in renal disease or arthritis. However, its role in corneal regeneration is less studied. In this study, filter disc of 2-mm diameter soaked in sodium hydroxide with a concentration of 0.5 N was placed at the center of the cornea for 30 s to establish a mouse model of corneal alkali injury. We found that 14-3-3zeta, which is mainly expressed in the epithelial layer, was upregulated following injury. Overexpression of 14-3-3zeta in ocular tissues via adeno-associated virus-mediated subconjunctival delivery promoted corneal wound healing, showing improved corneal structure and transparency. In vitro studies on human corneal epithelial cells showed that 14-3-3zeta was critical for cell proliferation and migration. mRNA-sequencing in conjunction with KEGG analysis and validation experiments revealed that 14-3-3zeta regulated the mRNA levels of ITGB1, PIK3R1, FGF5, PRKAA1 and the phosphorylation level of Akt, suggesting the involvement of the PI3K-Akt pathway in 14-3-3zeta-mediated tissue repair. 14-3-3zeta is a potential novel therapeutic candidate for treating severe corneal injury.

Congenital insensitivity to pain associated with PRDM12 mutation: Two case reports and a literature review.

Background: PRDM12 is a newly discovered gene responsible for congenital insensitivity to pain (CIP). Its clinical manifestations are various and not widely known. Methods: The clinical data of two infants diagnosed with CIP associated with PRDM12 mutation were collected. A literature review was performed, and the clinical characteristics of 20 cases diagnosed with a mutation of PRDM12 were summarized and analyzed. Results: Two patients had pain insensitivity, tongue and lip defects, and corneal ulcers. The genomic analysis results showed that variants of PRDM12 were detected in the two families. The case 1 patient carried heterozygous variations of c.682+1G > A and c.502C > T (p.R168C), which were inherited from her father and mother, respectively. We enrolled 22 patients diagnosed with CIP through a literature review together with our cases. There were 16 male (72.7%) and 6 female (27.3%) patients. The age of onset ranged from 6 months to 57 years. The prevalence of clinic manifestation was 14 cases with insensitivity to pain (63.6%), 19 cases with self-mutilation behaviors (86.4%), 11 cases with tongue and lip defects (50%), 5 cases with mid-facial lesions (22.7%), 6 cases with distal phalanx injury (27.3%), 11 cases of recurrent infection (50%), 3 cases (13.6%) with anhidrosis, and 5 cases (22.7%) with global developmental delay. The prevalence of ocular symptoms was 11 cases (50%) with reduced tear secretion, 6 cases (27.3%) with decreased corneal sensitivity, 7 cases (31.8%) with disappeared corneal reflexes, 5.5 cases (25%, 0.5 indicated a single eye) with corneal opacity, 5 cases (22.7%) with corneal ulceration, and 1 case (4.5%) with a corneal scar. Conclusion: The syndrome caused by PRDM12 mutation is a clinically distinct and diagnosable disease that requires joint multidisciplinary management to control the development of the disease and minimize the occurrence of complications.

Subconjunctival Administration of Mesenchymal Stem Cells Alleviates Ocular Inflammation in a Murine Model of Corneal Alkali Burn.

Corneal alkali burns cause extensive damage not only to the cornea but also to the intraocular tissues. As an anti-inflammatory therapy, subconjunctival administration of mesenchymal stem cells (MSCs) for corneal protection after corneal alkali burn has been explored. Little evidence demonstrates the potential of subconjunctival MSCs delivery in protecting the post-burn intraocular tissues. This study aimed to evaluate the therapeutic efficacy of subconjunctival injection of human placental (hP)-MSCs in protecting against ocular destruction after the burn. hP-MSCs were subconjunctivally administered to C57/BL mice after corneal alkali burn. Western blot of iNOS and CD206 was performed to determine the M1 and M2 macrophage infiltration in the cornea. Infiltration of inflammatory cells in the anterior uvea and retina was analyzed by flow cytometry. The TUNEL assay or Western blot of Bax and Bcl2 was used to evaluate the anti-apoptotic effects of MSCs. MSCs could effectively facilitate cornea repair by suppressing inflammatory cytokines IL-1ß, MCP-1, and MMP9, and polarizing CD206 positive M2 macrophages. Anterior uveal and retinal inflammatory cytokines expression and inflammatory cell infiltration were inhibited in the MSC-treated group. Reduced TUNEL positive staining and Bax/Bcl2 ratio indicated the anti-apoptosis of MSCs. MSC-conditioned medium promoted human corneal epithelial cell proliferation and regulated LPS-stimulated inflammation in RAW 264.7 macrophages, confirming the trophic and immunoregulatory effects of MSCs. Our findings demonstrate that subconjunctival administration of MSCs exerted anti-inflammatory and anti-apoptotic effects in the cornea, anterior uvea, and retina after corneal alkali burn. This strategy may provide a new direction for preventing post-event complications after corneal alkali burn.

[Effect of Bcl-2 antisense phosphorothioate oligodeoxynucleotides on bile duct carcinoma cell line QBC939].

OBJECTIVE: To investigate the effect of Bcl-2 antisense oligodeoxynucleotides (ASODN) on the cell proliferation and Bcl-2 expression in bile duct carcinoma cell line QBC939. METHODS: Bcl-2 ASODN and control sequence were transfected into cell line QBC939 by Lipofectamine 2000. The changes of Bcl-2 protein were detected by Western blot. The survival rate and colony formation rate of QBC939 cells incubating with Bcl-2 ASODN were evaluated by trypan blue staining assay and colony forming test. RESULTS: The densitometric analysis of Gel-photograph showed that the level of Bcl-2 protein expression in the ASODN transfected group was significantly lower than that in the controls (P < 0.01). Both trypan blue staining assay and colony forming test demonstrated that Bcl-2 ASODN could partially inhibit the growth of QBC939 cells. After incubating with Bcl-2 ASODN, the survival rate and colony formation rate of QBC939 cells were significantly lower than those of the controls (P < 0.05). CONCLUSION: Bcl-2 ASODN inhibits the cell proliferation in bile duct carcinoma cell line QBC939 by blocking the expression of bcl-2 gene.