Cyclic guanosine monophosphate-adenosine monophosphate synthetase/stimulator of interferon genes signaling aggravated corneal allograft rejection through neutrophil extracellular traps.

The activation of innate immunity following transplantation has been identified as a crucial factor in allograft inflammation and rejection. However, the role of cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS)/stimulator of interferon genes (STING) signaling-mediated innate immunity in the pathogenesis of allograft rejection remains unclear. Utilizing a well-established murine model of corneal transplantation, we demonstrated increased expression of cGAS and STING in rejected-corneal allografts compared with syngeneic (Syn) and normal (Nor) corneas, along with significant activation of the cGAS/STING pathway, as evidenced by the enhanced phosphorylation of TANK-binding kinase 1and interferon regulatory factor 3. Pharmacological and genetic inhibition of cGAS/STING signaling markedly delayed corneal transplantation rejection, resulting in prolonged survival time and reduced inflammatory infiltration. Furthermore, we observed an increase in the formation of neutrophil extracellular traps (NETs) in rejected allografts, and the inhibition of NET formation through targeting peptidylarginine deiminase 4 and DNase I treatment significantly alleviated immune rejection and reduced cGAS/STING signaling activity. Conversely, subconjunctival injection of NETs accelerated corneal transplantation rejection and enhanced the activation of the cGAS/STING pathway. Collectively, these findings demonstrate that NETs contribute to the exacerbation of allograft rejection via cGAS/STING signaling, highlighting the targeting of the NETs/cGAS/STING signaling pathway as a potential strategy for prolonging allograft survival.

Comparison of therapeutic effects between big-bubble deep anterior lamellar keratoplasty and penetrating keratoplasty for medically unresponsive Acanthamoeba keratitis.

PURPOSE: To compare the outcomes of big-bubble deep anterior lamellar keratoplasty (BB-DALK) and penetrating keratoplasty (PKP) in the management of medically unresponsive Acanthamoeba keratitis (AK). METHODS: This retrospective study included 27 eyes of BB-DALK and 24 eyes of PKP from a tertiary ophthalmology care centre. Glucocorticoid eye drops were subsequently added to the treatment plan 2 months postoperatively based on the evaluation using confocal laser scanning microscopy. The clinical presentations, best-corrected visual acuity (BCVA), postoperative refractive outcomes, graft survival, and Acanthamoeba recurrence were analyzed. RESULTS: The AK patients included in the study were in stage 2 or stage 3, and the percentage of patients in stage 3 was higher in the PKP group (P = 0.003). Clinical presentations were mainly corneal ulcers and ring infiltrates, and endothelial plaques, hypopyon, uveitis and glaucoma were more common in the PKP group (P = 0.007). The BCVA and the graft survival rate showed no statistically significant differences between the two groups at 1 year after surgery. However, 3 years postoperatively, the BCVA of 0.71 ± 0.64 logMAR, the graft survival rate of 89.5%, and the endothelial cell density of 1899 ± 125 cells per square millimeter in the BB-DALK group were significantly better than those of the PKP group (P = 0.010, 0.046, and 0.032, respectively). 3 eyes (11.1%) in the BB-DALK group and 2 eyes (8.3%) in the PKP group experienced Acanthamoeba recurrence, but the rates showed no statistically significant difference between the two groups (P = 1.000). In the PKP group, immune rejection and elevated intraocular pressure were observed in 5 and 6 eyes, respectively. CONCLUSION: Corneal transplantation is recommended for AK patients unresponsive to antiamoebic agents. The visual acuity and graft survival can be maintained after BB-DALK surgery. Acanthamoeba recurrence is not related to the surgical approach performed, whereas complete dissection of the infected corneal stroma and delayed prescribing of glucocorticoid eye drops were important to prevent recurrence.

Effects of femtosecond laser-assisted minimally invasive lamellar keratoplasty (FL-MILK) on mild-to-moderate and advanced keratoconus.

PURPOSE: To compare the outcomes of femtosecond laser-assisted minimally invasive lamellar keratoplasty (FL-MILK) for mild-to-moderate keratoconus (KC) and advanced KC. METHODS: Prospective case series study. Sixty-three eyes of 56 patients with progressive KC underwent FL-MILK were divided into group 1 [mean keratometry (Kmean) ≤ 53D] and group 2 (Kmean > 53D). Best spectacle-corrected visual acuity (BSCVA), Kmean, maximum keratometry (Kmax), anterior central corneal elevation (ACE), stiffness parameter A1 (SP-A1) and deformation amplitude (DA) were evaluated preoperatively and up to 24 months postoperatively. RESULTS: Mean BSCVA improved from 0.34 ± 0.13 logMAR preoperatively to 0.25 ± 0.13 logMAR at 24 months postoperatively in group 1 (F = 10.10, P < .0001), and from 0.54 ± 0.31 logMAR to 0.40 ± 0.26 logMAR (F = 9.06, P = .0002) in group 2. Group 2 showed an average Kmax reduction of 10.9 D and an average Kmean reduction of 3.9 D at 24 months postoperatively (both P < .0001), whereas no significant change was observed in group 1. Average ACE decreased from 19.2 ± 10.0 to 5.2 ± 8.4 at 24 months postoperatively in group 1 (F = 28.5, P < .0001), and from 46.2 ± 16.3 to 19.1 ± 9.0 (F = 49.6, P < .0001) in group 2; SP-A1 increased from 53.8 ± 12.7 mmHg/mm to 95.9 ± 20.2 mmHg/mm in group 1 (F = 70.0, P < .0001), and from 38.6 ± 13.4 mmHg/mm to 89.3 ± 18.2 mmHg/mm (F = 96.9, P < .0001) in group 2; DA decreased from 1.30 ± 0.14 mm to 1.17 ± 0.13 mm in group 1 (F = 14.0, P < .0001), and from 1.40 ± 0.16 mm to 1.18 ± 0.10 mm (F = 27.6, P < .0001) in group 2. CONCLUSIONS: FL-MILK can stabilize progressive KC in mild-to-moderate cases and advanced cases at 24-month follow-up. Steeper corneas are more likely to undergo flattening after FL-MILK. CLINICAL TRIAL: Date of registration: July 16, 2017. The title of the trail: www. CLINICALTRIALS: gov Trial registration number: NCT03229239. The name of the trial registry: ClinicalTrials.gov.

Microbiological Characteristics and Risk Factors Involved in Progression from Fungal Keratitis with Hypopyon to Keratitis-Related Endophthalmitis.

PURPOSE: To investigate the differences in microbiological characteristics, risk factors, drug resistance, and visual outcomes in three infections: fungal keratitis with hypopyon (FKH), keratitis-related fungal endophthalmitis (FKE), and fungal endophthalmitis without keratitis (FE). METHODS: An analytical cross-sectional study. RESULTS: In total, 14.57% of eyes with FKH progressed to endophthalmitis. Hypopyon, pre-existence of lens problems, topical steroid use and sever keratitis were significantly associated with the development of FKE. The risk factors of the FKH and FE group were mainly plant trauma and open globe trauma, respectively. Keratitis-related endophthalmitis (FKE) showed a significantly higher resistance than the other two groups. The FKH group had the best final visual acuity, while the FKE group had the worst. CONCLUSION: Hypopyon height, pre-existing lens problems, topical steroid use and sever keratitis are risk factors for progression to endophthalmitis in eyes with fungal keratitis, and its progression is not affected by a single fungus. The antifungal drugs resistance in patients with endophthalmitis related to keratitis was significantly higher than that associated with other reasons. Timely diagnosis and risk factor assessment are essential for ensuring early treatment of FKE.

Enhanced autophagy alleviated corneal allograft rejection via inhibiting NLRP3 inflammasome activity.

Autophagy has been reported to be involved in many aspects of innate and adaptive immunity. Manipulating autophagy is recognized as a promising therapeutic approach for treating immunological diseases, including allograft rejection, and graft-versus-host disease. However, whether autophagy was closely associated with the pathogenesis of corneal allograft rejection remains largely unknown. Here, we showed that rapamycin (RAPA)-induced autophagy alleviated corneal allograft rejection. By contrast, blocking autophagic activity using 3-methyladeine (3-MA) aggravated corneal transplantation rejection. Mechanistically, we revealed that the enhanced autophagic turnover by RAPA inhibited NLRP3 inflammasome activity through NLRP3 degradation. While blocking the fusion of autophagosomes with lysosomes by bafilomycin A1(BafA1), the reduced NLRP3 inflammasome activity induced by RAPA was significantly restored, with increased protein levels of NLRP3 and cleaved Casp-1(p10), as well as IL-1ß secretion. Moreover, we further revealed that pharmacologically blocking NLRP3 inflammasome signaling prolonged the survival of corneal allografts. Taken together, these findings underscored the critical roles of enhanced autophagy in treating corneal allograft rejection, which provided an alternative intervention strategy to control corneal transplantation rejection.

Risk factors and microbiological characteristics: from bacterial keratitis with hypopyon to keratitis-related endophthalmitis.

PURPOSE: To compare the clinical and microbiological characteristics in patients with bacterial keratitis with hypopyon (BKH), bacterial keratitis-related endophthalmitis (BKE), and bacterial endophthalmitis without keratitis (BE). METHODS: Data from all inpatients who were clinically diagnosed with BKH, BKE, and BE from 2018 to 2020 were collected retrospectively. The demographics, predisposing risk factors, clinical characteristics, microbiological profiles, and antibiotic susceptibility of the patients were evaluated. RESULTS: Approximately 9.46% (28/296) of eyes with BKH progressed to endophthalmitis. The hypopyon (OR = 5.35, 95% CI: 2.17-7.08) and corneal perforation (OR = 2.47, 95% CI: 1.04-4.86) were significantly related to the development of BKE. The odds ratios for hypopyon of less than 1 mm, 1-3 mm, and greater than 3 mm were 1, 2.09 (95% CI: 1.17-3.15), and 4.12 (95% CI:2.59-5.68), respectively. The predominant causative pathogen was Staphylococcus epidermidis (36.43%, 38.89%), followed by Streptococci (14.73%, 16.67%), Staphylococcus aureus (8.53%, 7.79%), and Pseudomonas aeruginosa (9.30%, 7.14%) in eyes with BKH and BE. However, the main pathogens were Pseudomonas aeruginosa (37.50%) and Staphylococcus aureus (31.25%) in eyes with BKE. In the BKH, BKE, and BE groups, almost 100% of Staphylococcus aureus isolates were sensitive to vancomycin (97.70%, 100%, 95.56%), about a half were sensitive to fluoroquinolones (51.85%, 39.90%, 62.34%), and approximately 30% were sensitive to trimethoprim/sulfa (27.77%, 21.56%, 33.56%) and cefazolin (41.47%, 20.31%, 38.81%). The susceptibility of Pseudomonas aeruginosa to fluoroquinolones antibiotics was 55.75%, 66.67%, and 62.58%, respectively, in the three groups. CONCLUSIONS: The height of hypopyon and corneal perforation are risk factors for progression to endophthalmitis in eyes with bacterial keratitis. When Staphylococcus aureus and Pseudomonas aeruginosa are identified, vigilance is required for advanced endophthalmitis.

Topical administration of tacrolimus and corticosteroids in tapering doses is effective in preventing immune rejection in high-risk keratoplasty: a 5-year follow-up study.

BACKGROUND: To evaluate the efficacy of the topical administration of immunosuppressants and corticosteroids in tapering doses in the management of patients with high-risk keratoplasty. METHODS: One hundred and six patients treated with topical immunosuppressants (50 eyes in the FK506 group and 56 eyes in the CsA group) and corticosteroid eye drops in tapering doses were enrolled in the study. The rates of rejection episodes, irreversible rejection, graft survival, and related influential factors were evaluated. RESULTS: The mean follow-up period was 48.1 ± 7.9 months (range, 36-60 months). The rates of rejection episodes and irreversible rejection were 14.0% and 6.00% in the FK506 group and 37.5% and 7.1% in the CsA group, respectively. Kaplan-Meier survival analysis demonstrated a significantly higher graft survival rate in the FK506 group (81.6%±5.3%, 71.1%±6.3%) compared with that in the CsA group (71.1%±6.3%, 57.5%±7.5%) at 3 and 5 years after surgery (P = 0.006). Multivariate logistic regression revealed that preoperative risk score ≥ 3 (P = 0.016) and endothelial immune rejection (P = 0.033) were risk factors associated with graft survival. CONCLUSIONS: Topical administration of tacrolimus and corticosteroids in tapering doses is effective in decreasing the incidence of immune rejection in high-risk keratoplasty. Careful instruction of patients on the reasonable use of topical tacrolimus is critical to avoid immune rejection induced by sudden discontinuation of medication.

Complex impact of DNA methylation on transcriptional dysregulation across 22 human cancer types.

Accumulating evidence has demonstrated that transcriptional regulation is affected by DNA methylation. Understanding the perturbation of DNA methylation-mediated regulation between transcriptional factors (TFs) and targets is crucial for human diseases. However, the global landscape of DNA methylation-mediated transcriptional dysregulation (DMTD) across cancers has not been portrayed. Here, we systematically identified DMTD by integrative analysis of transcriptome, methylome and regulatome across 22 human cancer types. Our results revealed that transcriptional regulation was affected by DNA methylation, involving hundreds of methylation-sensitive TFs (MethTFs). In addition, pan-cancer MethTFs, the regulatory activity of which is generally affected by DNA methylation across cancers, exhibit dominant functional characteristics and regulate several cancer hallmarks. Moreover, pan-cancer MethTFs were found to be affected by DNA methylation in a complex pattern. Finally, we investigated the cooperation among MethTFs and identified a network module that consisted of 43 MethTFs with prognostic potential. In summary, we systematically dissected the transcriptional dysregulation mediated by DNA methylation across cancer types, and our results provide a valuable resource for both epigenetic and transcriptional regulation communities.

Femtosecond laser-assisted minimally invasive lamellar keratoplasty for the treatment of advanced keratoconus.

BACKGROUND: To evaluate the initial safety and efficacy of femtosecond laser-assisted minimal invasive lamellar keratoplasty (FL-MILK) for advanced keratoconus. METHODS: Twenty-two patients (22 eyes) with advanced keratoconus were included in this prospective study. All the involved eyes underwent FL-MILK. The femtosecond laser was used to create an intrastromal pocket with a 2.3 mm incision in the recipient cornea. Then a stromal button with a diameter of 9.0 mm and a depth of 200 µm was gently inserted into the intrastromal pocket through the 2.3 mm incision and flattened. No sutures were applied. Follow-up was conducted for 24 months. RESULTS: Twenty-two patients completed follow-up data for 12 months, 16 patients had 24 months follow-up. No epithelial implantation, infection or allogeneic rejection were observed during the follow-up. Based on baseline values, postoperative 12 months values and postoperative 24 months values, clinical significantly improvement was recorded in corrected distance visual acuity (CDVA) (0.40 ± 0.18 logMAR vs. 0.30 ± 0.12 logMAR and 0.23 ± 0.13 logMAR), the anterior central corneal elevation (29.14 ± 15.33 µm vs. 14.45 ± 13.75µm and 11.38 ± 8.33 µm), and corneal higher-order aberrations (3.536 ± 1.503 vs. 2.761 ± 1.517 and 0.994 ± 0.391). Corneal biomechanical properties in all eyes improved significantly. SP-A1 increased from 48.64 ± 12.87 preoperatively to 87.26 ± 21.01 postoperative 12 months and 88.77 ± 18.26 postoperative 24 months; deformation amplitude (DA) decreased from 1.36 ± 0.15 preoperatively to 1.21 ± 0.12 postoperative 12 months and 1.19 ± 0.19 postoperative 24 months. CONCLUSIONS: Initial experience suggests that this minimally invasive transplantation may be a feasible option for advanced keratoconus. A larger cohort and longer follow-up are required to validate our results and establish long-term safety and efficacy of the procedure.

Clinical Characteristics and Outcomes of Rare Fungal Keratitis Caused by Verticillium dahliae.

PURPOSE: To observe clinical characteristics and treatment outcomes of fungal keratitis caused by Verticillium dahliae. METHODS: Clinical data of 7 patients diagnosed as fungal keratitis cause by Verticillium dahliae were retrospectively analyzed. The clinical manifestations, mycology, in vitro antifungal susceptibility, treatment regimens and prognoses of the patients were evaluated. RESULTS: All 7 patients were farm worker, of which 5 cases were caused by plant trauma. The corneal ulcer had a round shape and a relatively limited range with the diameters mainly in the range of 2-7 mm. The stromal infiltration was mild, and had no pseudopodia, mossiness or endothelial plaques. Intact hyphaes were detected in corneal scrapings and confocal microscopy, isolates were identified by morphology and by sequencing the internal transcribed spacer region of ribosomal DNA. In vitro antifungal susceptibility testing showed that the most sensitive antifungal drug was Amphotericin B. In the 6 patients with an ulcer less than 2/3 of the corneal thickness, the ulcer healed after 18 days of antifungal treatment only in one eye. The other five patients underwent corneal ulcer debridement or conjunctival flap covering surgery. The remaining one patient with ulcer depth more than 2/3 of the corneal thickness underwent lamellar keratoplasty. CONCLUSION: Fungal keratitis caused by Verticillium dahliae has typical signs of a mild inflammatory response, and is not sensitive to antifungal drugs. It is recommended that patients undergo corneal ulcer debridement as soon as possible to promote rapid healing of the ulcers.

Research Progress on the Positive and Negative Regulatory Effects of Rhein on the Kidney: A Review of Its Molecular Targets.

Currently, both acute kidney injury (AKI) and chronic kidney disease (CKD) are considered to be the leading public health problems with gradually increasing incidence rates around the world. Rhein is a monomeric component of anthraquinone isolated from rhubarb, a traditional Chinese medicine. It has anti-inflammation, anti-oxidation, anti-apoptosis, anti-bacterial and other pharmacological activities, as well as a renal protective effects. Rhein exerts its nephroprotective effects mainly through decreasing hypoglycemic and hypolipidemic, playing anti-inflammatory, antioxidant and anti-fibrotic effects and regulating drug-transporters. However, the latest studies show that rhein also has potential kidney toxicity in case of large dosages and long use times. The present review highlights rhein's molecular targets and its different effects on the kidney based on the available literature and clarifies that rhein regulates the function of the kidney in a positive and negative way. It will be helpful to conduct further studies on how to make full use of rhein in the kidney and to avoid kidney damage so as to make it an effective kidney protection drug.

Comparison of complications and visual outcomes between big-bubble deep anterior lamellar keratoplasty and penetrating keratoplasty for fungal keratitis.

BACKGROUND: To compare the postoperative complications and visual outcomes of big-bubble deep anterior lamellar keratoplasty (BB-DALK) and penetrating keratoplasty (PK) for fungal keratitis (FK). METHODS: This retrospective study included 94 cases of BB-DALK for FK and 161 cases of PK for FK from a tertiary ophthalmology care centre. RESULTS: The most common FK pathogens were Fusarium (n = 84, 32.9%) and Aspergillus (n = 67, 26.3%). The recurrence rates after BB-DALK and PK were 3.2 and 5%, respectively (p = 0.723). The follow-up duration was 31.9 ± 15.8 months in the BB-DALK group and 33.9 ± 15.0 months in the PK group. The immune rejection rate was significantly lower in the BB-DALK group than in the PK group (1.1 vs. 18.6%, p < 0.001), as was the incidence of secondary glaucoma (p = 0.018). Endothelial cell density in the BB-DALK group tended to be stable at postoperative month 6, whereas the PK group still attenuated at a hyper-physiological rate. Postoperative best-corrected visual acuity (BCVA) significantly improved in both groups (p < 0.001). No significant difference between-group was observed in BCVA, refractive cylinder, and spherical equivalent postoperatively. CONCLUSION: Big-bubble DALK is a useful and safe alternative to PK for medically uncontrolled FK.

Clinical outcomes of double continuous suture in femtosecond laser-assisted lamellar keratoplasty for keratoconus.

To assess the surgical outcomes in patients who underwent femtosecond laser-assisted lamellar keratoplasty with double continuous suture for keratoconus, 100 patients (102 eyes) with keratoconus in advanced stages undergoing femtosecond laser-assisted lamellar keratoplasty in Shandong Eye Hospital were studied. In the management of keratoconus, 50 patients (52 eyes) received double continuous suture, and 50 patients (50 eyes) underwent interrupted suture. The follow-up duration was 1 year. Best-corrected visual acuity (BCVA), corneal astigmatism, cosmetic outcomes, and surgical complications were measured as outcome indicators. The epithelium healed at 3 ± 2 days and 4 ± 2 days in the double continuous suture group and the interrupted suture groups, respectively (P > 0.05). At 6 months after surgery, the average visual acuity was 20/125 and 20/100 (P > 0.05), and the average BCVA was 20/32 and 20/40 (P > 0.05), respectively. At 1 year after surgery, the average visual acuity was 20/63 and 20/80 (P > 0.05), and the average BCVA was 20/32 and 20/25 (P > 0.05), respectively; the mean curvature was 43.24 ± 5.15 D and 43.31 ± 5.58 D (P > 0.05), the mean astigmatism was 3.21 ± 1.74 D and 5.35 ± 1.37 D (P < 0.05).The looseness of sutures were found in 2 patients and 15 patients in both groups, respectively (P < 0.05). No postoperative infection or immune rejection occurred in either group during the follow-up. Comparing with the interrupted suture, using the continuous suture in femtosecond laser-assisted lamellar keratoplasty for keratoconus markedly limited the looseness of sutures with lesser corneal astigmatism and better visual quality postoperative.

Rare Fungal Keratitis Caused by Coprinellus Radians.

A case of fungal keratitis due to Coprinellus radians is reported. To our knowledge, fungal keratitis caused by this species was rare. Fungal hyphae were detected in corneal scrapings, and isolates were identified by morphology and by sequencing the internal transcribed spacer region of ribosomal DNA. The patient was treated with systemic and local antifungal therapy for 5 days, and lamellar keratoplasty was performed after no obvious improvement in symptoms. The in vitro antifungal susceptibilities of the case strain were tested for six antifungal agents. The results showed that 5-fluorouracil was resistant, fluconazole was moderately sensitive, and the other drugs assayed (amphotericin B, posaconazole, itraconazole and voriconazole) were highly effective against this fungus.

Comparison of the efficacy of different cell sources for transplantation in total limbal stem cell deficiency.

PURPOSE: The purpose of this study was to compare the efficacy of allogeneic cultured limbal epithelial transplantation (ACLET) and cultivated oral mucosal epithelial transplantation (COMET) in treating total limbal stem cell deficiency (LSCD). METHODS: In this retrospective cohort study, 73 patients (76 eyes) with total LSCD, including 41 patients (42 eyes) treated with ACLET and 32 patients (34 eyes) receiving COMET, were evaluated. The age, gender and injury cause of all patients were recorded. RESULTS: The mean follow-up was 23.3 ± 9.9 months in the ACLET group and 16.1 ± 5.8 months in the COMET group. A higher incidence of persistent epithelial defect was observed after COMET (P = 0.023). The overall ocular surface grading scores were all lower in the ACLET group than in the COMET group at 3, 6, and 12 months after surgery and the last follow-up. Kaplan-Meier survival curve analysis demonstrated a significantly higher success rate of ACLET (71.4%), compared with that of COMET (52.9%; P = 0.043). The risk of graft failure was higher in patients with entropion and trichiasis, incomplete eyelid closure and treated with COMET. The graft failure risk rate after COMET was 3.5 times higher than that of ACLET. CONCLUSIONS: For total LSCD patients, ACLET should be prioritized, since limbal epithelial cells have better ability to maintain corneal epithelial integrity and ocular surface stability and benefit the ocular surface when compared with oral mucosal epithelial cells. Preoperative and postoperative eyelid abnormalities should be corrected as early as possible.

Simultaneous determination of phenolic compounds in Equisetum palustre L. by ultra high performance liquid chromatography with tandem mass spectrometry combined with matrix solid-phase dispersion extraction.

A method based on matrix solid-phase dispersion extraction followed by ultra high performance liquid chromatography with tandem mass spectrometry is presented for the extraction and determination of phenolic compounds in Equisetum palustre. This method combines the high efficiency of matrix solid-phase dispersion extraction and the rapidity, sensitivity, and accuracy of ultra high performance liquid chromatography with tandem mass spectrometry. The influential parameters of the matrix solid-phase dispersion extraction were investigated and optimized. The optimized conditions were as follows: silica gel was selected as dispersing sorbent, the ratio of silica gel to sample was selected to be 2:1 (400/200 mg), and 8 mL of 80% methanol was used as elution solvent. Furthermore, a fast and sensitive ultra high performance liquid chromatography with tandem mass spectrometry method was developed for the determination of nine phenolic compounds in E. palustre. This method was carried out within <6 min, and exhibited satisfactory linearity, precision, and recovery. Compared with ultrasound-assisted extraction, the proposed matrix solid-phase dispersion procedure possessed higher extraction efficiency, and was more convenient and time saving with reduced requirements on sample and solvent amounts. All these results suggest that the developed method represents an excellent alternative for the extraction and determination of active components in plant matrices.

Involvement of cGAS/STING Signaling in the Pathogenesis of Candida albicans Keratitis: Insights From Genetic and Pharmacological Approaches.

Purpose: Fungal keratitis (FK) is an invasive corneal infection associated with significant risk to vision. Although the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) signaling pathway has been recognized for its role in defending against viral infections, its involvement in FK still remains largely unclear. This study sought to elucidate the contribution of the cGAS/STING signaling pathway to the pathogenesis of FK. Methods: The expression of cGAS/STING signaling components was assessed in a murine model of Candida albicans keratitis through RNA sequencing, western blot analysis, immunofluorescence staining, and real-time PCR. Both genetic (utilizing Sting1gt/gt mice) and pharmacological (using C176) interventions were employed to inhibit STING activity, allowing for the evaluation of resultant pathogenic alterations in FK using slit-lamp examination, clinical scoring, hematoxylin and eosin (H&E) staining, fungal culture, and RNA sequencing. Subconjunctival administration of the NOD-like receptor protein 3 (NLRP3) inflammasome inhibitor MCC950 was performed to evaluate FK manifestations following STING activity blockade. Furthermore, the impact of the STING agonist diABZI on FK progression was investigated. Results: Compared to uninfected corneas, those infected with C. albicans exhibited increased expression of cGAS/STING signaling components, as well as its elevated activity. Inhibiting cGAS/STING signaling exacerbated the advancement of FK, as evidenced by elevated clinical scores, augmented fungal load, and heightened inflammatory response, including NLRP3 inflammasome activation and pyroptosis. Pharmacological inhibition of the NLRP3 inflammasome effectively mitigated the exacerbated FK by suppressing STING activity. Conversely, pre-activation of STING exacerbated FK progression compared to the PBS control, characterized by increased fungal burden and reinforced inflammatory infiltration. Conclusions: This study demonstrates the essential role of the cGAS/STING signaling pathway in FK pathogenesis and highlights the necessity of its proper activation for the host against FK.

Characteristics of immune rejection after allogeneic cultivated limbal epithelial transplantation.

PURPOSE: To investigate the characteristics and prognosis of immune rejection after allogeneic cultivated limbal epithelial transplantation (CLET). DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Forty-two eyes of 41 patients undergoing allogeneic CLET for total limbal stem cell deficiency who completed a follow-up of at least 12 months. METHODS: Allogeneic cultivated limbal epithelial cells using human amniotic membrane as a carrier were transplanted into the recipient eye. Immune rejection occurred in 10 eyes; the medical records of these patients were reviewed. MAIN OUTCOME MEASURES: Best-corrected visual acuity, corneal opacification and neovascularization, immunofluorescence staining of CD4 and CD8 T cells, and distribution of Langerhans cells (LCs) in the corneal epithelium. RESULTS: The corneal epithelium became edematous with epithelial rejection lines and peripheral epithelial defects in 6 eyes. Circumlimbal vessels were hyperemic and extended up to the corneal stroma, thus resulting in aggravation of corneal neovascularization in 8 eyes. Corneal stromal opacification was observed in 9 eyes. CD4(+) or CD8(+) T cells were detected in 5 of 6 eyes from which impression cytology specimens were obtained. In vivo confocal microscopy examination revealed an accumulation of LCs in the central and peripheral corneal epithelium. All patients responded to antirejection therapy. One eye developed a second episode. CONCLUSIONS: The diagnosis of immune rejection after allogeneic CLET mainly depends on typical clinical manifestations. Delayed recognition can result in worsening corneal opacification and neovascularization. Reasonable use of topical immunosuppressives and a close follow-up within 6 months after allogeneic CLET are critical to improve the prognosis. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Comparison of penetrating keratoplasty and deep lamellar keratoplasty for macular corneal dystrophy and risk factors of recurrence.

PURPOSE: To compare the therapeutic effects of penetrating keratoplasty (PK) and deep anterior lamellar keratoplasty (DALK) on patients with macular corneal dystrophy (MCD) and to analyze the risk factors of postoperative recurrence. DESIGN: Retrospective, interventional, comparative case series. PARTICIPANTS: Fifty-one patients (78 eyes) with MCD treated by PK or DALK at Shandong Eye Institute between January 1992 and December 2010. METHODS: The medical records of the patients were reviewed retrospectively. MAIN OUTCOME MEASURES: Best-corrected visual acuity, corneal endothelial density, complications, recurrence, graft survival, and risk factors for recurrence. RESULTS: Penetrating keratoplasty was performed in 57 eyes, and DALK was performed in 21 eyes. The mean follow-up time was 5.1±4.1 years (range, 1.0-18.0 years). The best-corrected visual acuity of the PK group was much better than that of the DALK group at 1, 2, 3, and 5 years. The corneal endothelial density was reduced to 1000 cells/mm2 or less within 5 years in 21.6% (11/51) of eyes treated by PK and in none of the eyes treated by DALK. The 1-year incidence rate of complications was 21.1% in the PK group, higher than the 4.8% rate in the DALK group. At the last visit, the rate of graft clarity was 87.7% and 85.7% in the 2 groups, respectively. Ten eyes (17.5%) treated by PK had recurrent MCD, with a rate of 0.8%, 7.7%, and 40% at 1, 5, and 10 years, respectively, whereas 9 eyes (42.9%) treated by DALK demonstrated recurrence, with a rate of 14.3% and 49.5% at 1 and 5 years, respectively. The recurrence risk was higher in patients whose age was 18 years or younger at onset or younger than 30 years at surgery. The recurrence risk after DALK was 5.066 times higher than that after PK. CONCLUSIONS: Penetrating keratoplasty more often immediately improves the visual acuity of patients with MCD, but many complications seem to be inevitable, especially continuous loss of corneal endothelium. Despite poor visual acuity and recurrence after surgery, DALK may produce fewer complications overall and more durable stability of the ocular surface compared with PK. The selection of PK or DALK for MCD should depend on the actual need and situation of certain patients. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Characterization and Evaluation of Rapamycin-Loaded Nano-Micelle Ophthalmic Solution.

Rapamycin-loaded nano-micelle ophthalmic solution (RAPA-NM) offers a promising application for preventing corneal allograft rejection; however, RAPA-NM has not yet been fully characterized. This study aimed to evaluate the physicochemical properties, biocompatibility, and underlying mechanism of RAPA-NM in inhibiting corneal allograft rejection. An optimized RAPA-NM was successfully prepared using a polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol (PVCL-PVA-PEG) graft copolymer as the excipient at a PVCL-PVA-PEG/RAPA weight ratio of 18:1. This formulation exhibited high encapsulation efficiency (99.25 ± 0.55%), small micelle size (64.42 ± 1.18 nm), uniform size distribution (polydispersity index = 0.076 ± 0.016), and a zeta potential of 1.67 ± 0.93 mV. The storage stability test showed that RAPA-NM could be stored steadily for 12 weeks. RAPA-NM also displayed satisfactory cytocompatibility and high membrane permeability. Moreover, topical administration of RAPA-NM could effectively prevent corneal allograft rejection. Mechanistically, a transcriptomic analysis revealed that several immune- and inflammation-related Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were significantly enriched in the downregulated genes in the RAPA-NM-treated allografts compared with the rejected allogenic corneal grafts. Taken together, these findings highlight the potential of RAPA-NM in treating corneal allograft rejection and other ocular inflammatory diseases.