Isolation and characterization of rabbit limbal niche cells.

Limbal niche cells (LNCs) are one of the most important supporting cells for corneal epithelial stem cells (CES), however, research on LNCs has been mostly limited to humans and rats previously. To expand the research work into the rabbit animal model, one of the most often used animals in stem cell study, this study was carried out for the in vitro isolation and identification of rabbit LNCs. Rabbit LNCs were isolated by collagenase A digestion method and single cells were obtained, the cells were then seeded on 5% Matrigel-coated plastic surface and cultured in modified embryonic stem cell medium (MESCM). Three biological replicates of the isolating and characterization were recorded from New Zealand White rabbits aged from 2.5 months to 5 months. LNC markers (VIM/CD90/CD105/SCF/PDGFRß) were analyzed using tyramide signal amplification (TSA) staining, immunohistochemical staining (IHC), western blotting (WB), and real-time reverse transcription polymerase chain reaction (qPCR). TSA staining suggested that VIM was highly expressed in rabbit limbus stroma, which was confirmed by WB, and P63α was expressed in the basal limbus epithelium. Pan-CK and CK12 were highly expressed in the central corneal epithelium but lightly expressed in the limbal epithelium. The WB result indicated that PDGFRß and VIM expressions in rabbit-LNCs P4 were higher than in P1 and P7. In addition, rabbit corneal epithelium highly expressed Paired Box 6 (PAX6) and Epidermal growth factor-like domain 6(EGFL6). For the three repeat experiments, the cell expansion activity of rabbit-LNC was highest at P4. Rabbit-LNCs were passaged from P0 to P7, and the number of cell doublings (NCD) of P4 for the three repeat experiments was 2.816, 2.737, and 2.849. qPCR showed that high mRNA expression levels of VIM, CD90, CD105, SCF, and PDGFRß in rabbit-LNCs P4. In conclusion, rabbit-LNCs could be successfully isolated by the collagenase A digestion method as used in human tissue. There were similar characteristics between rabbit and human LNCs (VIM+/CD90+/CD105+/SCF+/PAX6+/PDGFRß+).

Effect of artificial tears on dynamic optical quality in patients with dry eye disease.

BACKGROUND: In clinical practice, fluctuating vision or decreased quality of vision is a common complaint in DED patients. Our study was designed to investigate the change in dynamic optical quality in dry eye patients after the use of artificial tears. METHODS: Fifty-nine patients with dry eye disease (DED) and 31 control subjects were included in this prospective case-control study. There was no significant difference in age and sex between these two groups (P = 0.342, P = 0.847, respectively). Clinical evaluation of the ocular surface included Ocular Surface Disease Index (OSDI), tear film break-up time (TBUT), lipid layer thickness (LLT), and Schirmer I test. DED patients were divided into two groups, mild (31 patients) and severe (28 patients). The optical quality of the tear film was measured with the Optical Quality Analysis System (OQAS) using the mean objective scatter index (mean OSI), standard deviation of objective scatter index (SD-OSI) and modulation transfer function cut-off (MTF cut-off). After baseline examinations, one drop of artificial tears (ATs, carboxymethylcellulose ophthalmic solution, 0.5%) was instilled in both eyes, and optical quality parameters were measured again at 5 and 30 min following application of ATs. RESULTS: At baseline, the mean OSI was higher in the DED group (0.95 ± 0.54) than in controls (0.54 ± 0.23, P < 0.001). The SD-OSI was also significantly increased in DED patients (0.44 ± 0.71) compared to control subjects (0.12 ± 0.06, P = 0.003). Five minutes after AT instillation, mean OSI and SD-OSI decreased significantly in severe DED patients (P = 0.044; P = 0.018), remained unchanged in mild DED patients, and increased in the control group (P = 0.019; P < 0.001). Thirty minutes after AT instillation, no significant difference in optical quality parameters was observed among the three groups. CONCLUSION: The effect of ATs on optical quality in patients with DED may differ according to the severity of the disease. Measurement of optical quality might be a promising tool to evaluate the effects of various ATs and possibly individualize treatment in DED patients.

Basic Fibroblast Growth Factor Supports the Function of Limbal Niche Cells via the Wnt/ß-Catenin Pathway.

Purpose: To test the effects and underlying mechanisms of basic fibroblast growth factor (bFGF) on the limbal niche cell (LNC) function ex vivo. Methods: By using different concentrations of bFGF (0, 4, 8, 12, and 16 ng/mL) and fibroblast growth factor receptor (FGFR) inhibitors, the effects of bFGF on LNC proliferation, expression of stem cell markers, and transcription levels of the ß-catenin were investigated. Single-cell RNA sequencing (scRNA-seq) was used to analyze the action and mechanisms of FGFR subtypes and the Wnt/ß-catenin pathway during LNC culture. An mature corneal epithelial cell (MCEC)/LNC three-dimensional model was constructed to verify whether bFGF activates the Wnt/ß-catenin pathway in LNC by inhibiting FGFR or ß-catenin targets. Results: scRNA-seq showed that FGFR1 is the main receptor in LNC, along with the molecules in the Wnt pathway, including WNT2, FZD7, LRP5, LRP6, and ß-catenin. The 12 ng/mL bFGF treatment group showed higher LNC proliferation rate and transcription levels of OCT4, SOX2, NANOG, and ß-catenin than any other groups (P < 0.001). In the MCEC/LNC co-culture model, MCEC/LNC treated with 12 ng/mL bFGF promoted the aggregation of the spheres than other groups, associated with increased transcription levels of P63α, WNT2, ß-catenin, and a decreased transcription level of CK12 (P < 0.001). Wnt/ß-catenin inhibitor LF3 treatment reversed the abovementioned effect of bFGF. Conclusions: bFGF could maintain and promote the stemness of LNC via the FGFR1/Wnt2/FZD7/LRP6 axis in a concentration-dependent manner.

Assessment of iodine nutritional status and gestational thyroid function reference ranges during the first trimester of pregnancy in Taiwan.

BACKGROUND: Iodine nutrition is critical for fetal neurodevelopment in the first trimester of pregnancy, a period associated with dramatic changes in thyroid function. The aim of this study was to evaluate iodine nutritional status and thyroid function reference ranges in the first trimester in Taiwan. METHODS: Pregnant women aged 20 years and above in the first trimester were recruited in Taipei Veterans General Hospital, Taiwan from March 2019 to July 2022. Each participant provided a spot urine sample for measurement of urinary iodine concentration (UIC) and a blood sample for checkup of thyroid function and thyroid autoantibodies. A simple food frequency questionnaire was also completed. RESULTS: A total of 209 women with a mean age of 32.9 ± 4.4 years were enrolled. The median UIC was 160.9 µg/L (interquartile range [IQR]: 105.0-246.2 µg/L), indicating overall iodine sufficiency. The gestational thyroid function reference ranges were: thyroid stimulating hormone (TSH) (median: 0.93 [0.007-2.9] µIU/mL), free T4 (1.3 [0.93-2.2] ng/dL), free T3 (3.0 [2.3-5.0] ng/dL), total T4 (9.9 [6.4-16.9] ng/dL), and total T3 (135 [88-231] ng/dL). If the nonpregnant reference range of serum TSH was used, eight women (4.8%) would be misclassified as having subclinical hyperthyroidism, and two women (1.2%) with subclinical hypothyroidism would be missed. In multivariate analysis, nulliparous (adjusted odds ratio [OR] from model 1-3: 2.02, 2.05, 2.02; 95% CI, 1.08-3.77, 1.10-3.81, 1.11-3.66; p = 0.027, 0.023, 0.022, respectively) and multivitamin nonusers (adjusted OR from model 1-3: 1.86, 1.85, 1.78; 95% CI, 1.04-3.34, 1.03-3.32, 1.004-3.71; p = 0.038, 0.039, 0.049, respectively) had increased odds of having lower UIC levels <150 µg/L. CONCLUSION: The iodine nutritional status in the first trimester is adequate in Taiwan; however, certain subgroups such as nulliparous and multivitamin nonusers are still at risk for iodine deficiency. Gestational thyroid function reference ranges are needed for correct diagnosis of thyroid dysfunction in pregnancy.

Colletotrichum keratitis: An important fungal infection of nine human eyes.

Colletotrichum is a relatively uncommon cause of human infection. Previous findings on Colletotrichum keratitis were scarce, and most diagnoses were based on morphological distinction, perhaps underestimating the incidence of Colletotrichum species. This research describes the clinical characteristics and treatment outcomes of 9 cases of Colletotrichum keratitis discovered in our hospital using next-generation sequencing (NGS).We reviewed 78 patients with NGS-proven fungal keratitis between September 1, 2021 and May 31, 2023, 9 patients (11.5 %) were verified as infected with Colletotrichum species, and their medical records were reviewed to identify the clinical characteristics. NGS revealed that 3 patients were infected with C. truncatum, 3 patients with C. gloeosporioides, and the other 3 patients with C. fructicola. Seven patients had a history of corneal plant trauma (all three patients with C. fructicola had corneal injury history due to chestnut burrs), one patient was infected by mosquitoes flying into the eye, and one patient had an unknown origin. Seven patients underwent penetrating keratoplasty, while two patients underwent lamellar keratoplasty. Eight patients healed after keratoplasty, but one required evisceration due to recurrence of fungal infection in the anterior chamber and intractable discomfort.In conclusion NGS allows for more precise diagnosis and enhances epidemiological awareness of Colletotrichum keratitis, which is not as rare as previously reported.

Low-Density Lipoprotein Cholesterol Is Not a Major Determinant of High-Sensitivity C-Reactive Protein Levels in People Without Known Diabetes Mellitus.

Background: It has been well established that high-sensitivity C-reactive protein (hs-CRP) is strongly associated with obesity, insulin resistance, high blood pressure, and dyslipidemia. However, the effects of different lipid parameters on hs-CRP levels are less deliberated. The purpose of the study was to compare the relative contribution of triglycerides (TGs), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) to the levels of hs-CRP. Methods: Three hundred seventy-eight subjects without known history of diabetes were recruited for the study. No concomitant antilipid or antidiabetes agents were allowed. Each subject received anthropometric measurements, fasting sampling for lipid profile and hs-CRP, and a 75-gram oral glucose tolerance test for the measurements of insulin resistance (surrogated by insulin sensitivity index ISI0,120). Results: Levels of hs-CRP levels were positively correlated with Log (TG) and negatively correlated with HDL-C in partial correlation after adjustments for confounding variables, but not with LDL-C. The hs-CRP levels in the three groups by tertiles of LDL-C were similar. Subsequently, we found that body mass index (first step), Log (ISI0,120) (second step), and Log (TG) (third step) independently predicted the variance of Log (hs-CRP) in stepwise multiple regression. However, both HDL-C and LDL-C failed to be entered into the models to explain Log (hs-CRP). Conclusions: Our data demonstrated that Log (TG) was a major lipid determinant of hs-CRP levels. The contribution of LDL-C to the levels of hs-CRP might be insignificant.

Analysis of the association between serum antiaging humoral factor klotho and cardiovascular disease potential risk factor apolipoprotein B in general population.

Cardiovascular disease (CVD) is a prevalent health issue, and various risk factors contribute to its development, including blood lipids, blood pressure, diabetes, smoking, and alcohol consumption. Apolipoprotein B (ApoB) is related to CVD. ApoB is present on the surface of low-density lipoprotein (LDL), and its cellular recognition and LDL uptake are mainly achieved through recognition. It plays a crucial role in the diagnosis and treatment of CVD. This study aims to investigate the relationship between Klotho and ApoB in the general population of the United States as the correlation between serum Klotho and apoB is currently unknown. These findings could potentially guide the development of future treatments for CVD. This study utilized data from the National Health and Nutrition Examination Survey (NHANES) collected between 2007 and 2016. A linear regression model and smooth curve fitting were conducted to analyze the relationship between serum Klotho and apoB. The results indicate a negative correlation between serum Klotho concentration and apoB concentration (ß = -71.7; 95% confidence interval [CI]: -120.8, -22.6; P = .005). After adjusting for confounding variables, the negative correlation between apoB concentration and serum Klotho concentration became more significant (ß = -91.8; 95% CI: -151.3, -32.2; P = .004). When apoB concentration was converted from a continuous variable to a categorical variable (tertiles: T1 <0.8 g/L; T2: ≥0.8 g/L to <1.0 g/L; T3: ≥1.0 g/L), the serum klotho level of participants in the highest tertile (≥1.0 g/L) was -44.8 pg/mL (95% CI: -86.3, -3.2; P = .040) lower than that in the lowest tertile (<0.8 g/L). The smooth curve fitting diagram revealed differences in the relationship between serum Klotho concentration and apoB among individuals with different CVD risk factors. This study demonstrates a significant negative correlation between serum Klotho concentration and apoB concentration, even after controlling for confounding factors. The findings suggest that serum Klotho and apoB may be involved in the development of CVD, and targeting these factors could be a potential approach for CVD prevention and treatment.

Urinary acrolein protein conjugates-to-creatinine ratio is positively associated with diabetic peripheral neuropathy in patients with type 2 diabetes mellitus.

Acrolein, an unsaturated aldehyde, plays a pathological role in neurodegenerative diseases. However, less is known about its effects on peripheral neuropathy. The aim of this study was to investigate the association of acrolein and diabetic peripheral neuropathy in patients with type 2 diabetes. We recruited 148 ambulatory patients with type 2 diabetes. Each participant underwent an assessment of the Michigan Neuropathy Screening Instrument Physical Examination. Diabetic peripheral neuropathy was defined as Michigan Neuropathy Screening Instrument Physical Examination score ≥ 2.5. Serum levels and urinary levels of acrolein protein conjugates were measured. Urinary acrolein protein conjugates-to-creatinine ratios were determined. Patients with diabetic peripheral neuropathy had significantly higher urinary acrolein protein conjugates-to-creatinine ratios than those without diabetic peripheral neuropathy (7.91, 95% CI: 5.96-10.50 vs 5.31, 95% CI: 4.21-6.68, P = 0.029). Logarithmic transformation of urinary acrolein protein conjugates-to-creatinine ratios was positively associated with diabetic peripheral neuropathy in univariate logistic analysis, and the association remained significant in multivariate analysis (OR = 2.45, 95% CI: 1.12-5.34, P = 0.025). In conclusion, urinary acrolein protein conjugates-to-creatinine ratio may act as a new biomarker for diabetic peripheral neuropathy in type 2 diabetes. The involvement of acrolein in the pathogenesis of diabetic peripheral neuropathy warrants further investigation.

Research progress of biomimetic materials in oral medicine.

Biomimetic materials are able to mimic the structure and functional properties of native tissues especially natural oral tissues. They have attracted growing attention for their potential to achieve configurable and functional reconstruction in oral medicine. Though tremendous progress has been made regarding biomimetic materials, significant challenges still remain in terms of controversy on the mechanism of tooth tissue regeneration, lack of options for manufacturing such materials and insufficiency of in vivo experimental tests in related fields. In this review, the biomimetic materials used in oral medicine are summarized systematically, including tooth defect, tooth loss, periodontal diseases and maxillofacial bone defect. Various theoretical foundations of biomimetic materials research are reviewed, introducing the current and pertinent results. The benefits and limitations of these materials are summed up at the same time. Finally, challenges and potential of this field are discussed. This review provides the framework and support for further research in addition to giving a generally novel and fundamental basis for the utilization of biomimetic materials in the future.

A unifying criterion of the boiling crisis.

We reveal and justify, both theoretically and experimentally, the existence of a unifying criterion of the boiling crisis. This criterion emerges from an instability in the near-wall interactions of bubbles, which can be described as a percolation process driven by three fundamental boiling parameters: nucleation site density, average bubble footprint radius and product of average bubble growth time and detachment frequency. Our analysis suggests that the boiling crisis occurs on a well-defined critical surface in the multidimensional space of these parameters. Our experiments confirm the existence of this unifying criterion for a wide variety of boiling surface geometries and textures, two boiling regimes (pool and flow boiling) and two fluids (water and liquid nitrogen). This criterion constitutes a simple mechanistic rule to predict the boiling crisis, also providing a guiding principle for designing boiling surfaces that would maximize the nucleate boiling performance.

Isolation and Identification of Limbal Niche Cells.

Here we report a standard procedure for the isolation and identification of limbal niche cells (LNCs). Limbus tissue obtained from an eye bank was used for LNCs isolation. The tissue was divided into 12 pieces under aseptic conditions and digested for 18 h at 37 °C in the cell culture incubator using collagenase A to obtain cell clusters with LNCs and limbal epithelial progenitor cells. The cell clusters were further digested for 15 min at 37 °C using 0.25% trypsin-EDTA to obtain single cells and then cultured in modified embryonic stem cell medium (MESCM) on a plastic surface coated with 5% Matrigel. Cells were passaged upon 70% confluence, and LNCs were identified using immunofluorescence, real-time quantitative PCR (qPCR), and flow cytometry. Primary LNCs were isolated and passaged more than 12 times. The proliferation activity of LNCs from P4 to P6 was the highest. LNCs expressed higher stem cell markers than BMMSCs (SCF, Nestin, Rex1, SSEA4, CD73, CD90, MSX1, P75NTR, and PDGFRß). Furthermore, results showed that P4 LNCs uniformly expressed VIM, CD90, CD105, and PDGFRß, but not Pan-CK, which could be used as a marker for the identification of LNCs. Flow cytometric analysis showed that approximately 95%, 97%, 92%, and 11% of LNCs expressed CD73, CD90, CD105, and SCF respectively, while they were 68%, 99%, 20%, and 3% in BMMSCs. The standard process for LNC isolation and identification could provide a reliable laboratory basis for the widespread use of LNCs.

3D printed zirconia used as dental materials: a critical review.

In view of its high mechanical performance, outstanding aesthetic qualities, and biological stability, zirconia has been widely used in the fields of dentistry. Due to its potential to produce suitable advanced configurations and structures for a number of medical applications, especially personalized created devices, ceramic additive manufacturing (AM) has been attracting a great deal of attention in recent years. AM zirconia hews out infinite possibilities that are otherwise barely possible with traditional processes thanks to its freedom and efficiency. In the review, AM zirconia's physical and adhesive characteristics, accuracy, biocompatibility, as well as their clinical applications have been reviewed. Here, we highlight the accuracy and biocompatibility of 3D printed zirconia. Also, current obstacles and a forecast of AM zirconia for its development and improvement have been covered. In summary, this review offers a description of the basic characteristics of AM zirconia materials intended for oral medicine. Furthermore, it provides a generally novel and fundamental basis for the utilization of 3D printed zirconia in dentistry.

Diagnosis of fungal keratitis caused by Nectria haematococca through next-generation sequencing: review of literature and report of three cases.

OBJECTIVES: Fungal keratitis (FK) is a kind of serious corneal infection and penetrating keratoplasty (PKP) is needed when medical therapy fails. Although Nectria haematococca is found as endophytes in the roots of some plant species, there has been no report of N. haematococca infection in human. METHODS: We reviewed 46 patients who underwent PKP due to FK in our hospital from July 2021 to December 2021, and there were three patients who had relapsed. The next-generation sequencing revealed that all three corneas were infected with N. haematococca. RESULTS: Based on the ocular manifestation and treatment course of three cases, we summarize the characteristics of N. haematococca FK: the scope of corneal infection was widespread with severe hypopyon. The effect of local use of fluconazole and voriconazole was not ideal, and PKP was the main treatment. Even after a large-scale corneal lesion resection, the lesion may recur. The recurrence occurred primarily in the second week after PKP. CONCLUSION: This is the first clinical report of N. haematococca infection in humans. Compared with the other currently known FK caused by the Fusarium solani species complex, N. haematococca keratitis is more severe and more likely to recur.

Application Prospect and Preliminary Exploration of GelMA in Corneal Stroma Regeneration.

Corneal regeneration has become a prominent study area in recent decades. Because the corneal stroma contributes about 90% of the corneal thickness in the corneal structure, corneal stromal regeneration is critical for the treatment of cornea disease. Numerous materials, including deacetylated chitosan, hydrophilic gel, collagen, gelatin methacrylate (GelMA), serine protein, glycerol sebacate, and decellularized extracellular matrix, have been explored for keratocytes regeneration. GelMA is one of the most prominent materials, which is becoming more and more popular because of its outstanding three-dimensional scaffold structure, strong mechanics, good optical transmittance, and biocompatibility. This review discussed recent research on corneal stroma regeneration materials and related GelMA.

Progress of EGFL6 in angiogenesis and tumor development.

The epidermal growth factor (EGF) superfamily includes the protein 6 with an epidermal growth factor-like protein (EGFL6). EGFL6 has a signal peptide domain with an amino terminus and a MAM domain with a carboxy terminus. There are four whole EGF-like repeat regions and one partial EGF-like repeat region. Three of these regions include calcium-binding structures and an arg-gly-asp (RGD) integrin interaction motif. The epidermal growth factor-like (EGFL) and EGF domains have identical amino acid residues. Cell division, differentiation, mortality, cell adhesion, and migration are all affected by EGFL6. EGFL proteins are involved in a broad range of biological activities, making it important in tumor development and angiogenesis. We highlighted the latest development of EGFL6 research on tumor proliferation, invasion, and migration in this review.

Thickness Measurement of Endothelium-Descemet Membrane in Descemt Membrane Detachment Patients Using High-Definition Optical Coherence Tomography.

Purpose: (1) To measure the corneal endothelium-Descemet membrane (EDM) layer thickness in Descemet membrane detachment (DMD) patients in vivo using high-definition optical coherence tomography (HD-OCT), and to investigate its correlation with age. (2) To explore whether the detachment time will affect the EDM thickness. (3) To explore whether the EDM thickness of cornea with DMD was different from that without DMD. Participants: Patients with DMD were divided into three groups. Group 1 included twenty-three patients whose Descemet membrane (DM) was partial or complete detached from the corneal stroma after various ocular surgeries. Group 2 included eight patients from group 1 who underwent twice HD-OCT examination on different days before the DM reattached to the stroma. Group 3 included nine patients from group 1 who had clear grayscale boundary between the DM and stroma in HD-OCT images after DM reattachment. Methods: All patients underwent HD-OCT and EDM thickness was measured using Image -Pro Plus 6.0. In Group 1, regression analyses were used to evaluate the correlation between EDM thickness and age, and the thickness difference between the ≤50-year-old group and the >50-year-old group was analyzed by independent sample t-test. In Group 2, paired samples t-test was used to check whether detachment time would affect EDM thickness. In Group 3, paired samples t-test was used to check whether the EDM thickness of cornea with DMD was different from that without DMD. p < 0.05 was considered significant. Results: In Group 1, the EDM thickness measured on the first post-operative day was 27.8 ± 3.6 µm, and a positive correlation was found between EDM thickness and age (r = 0.619, p < 0.05). The EDM thickness of ≤50-year-old group and >50-year-old group were 23.9 ± 3.2 and 29.2 ± 2.6 µm, and there was a significant difference between the two groups (p = 0.001). In Group 2, the first measurement of EDM thickness was 27.5 ± 4.0 µm, the second measurement was 27.6 ± 4.2 µm, the interval between the two measurements was 2.1 ± 1.6 days, and there was no significant difference between the two measurements (p = 0.328). In Group 3, the EDM thickness with DM detachment was 28.3 ± 3.5 µm, with DM reattachment was 23.4 ± 2.4 µm, there was a significant difference between the two measurements (p = 0.002). Conclusions: The EDM thickness in the state of DMD is thicker than its actual thickness in normal cornea, and EDM thickness of the >50-year-old group is much thicker than that of the ≤50-year-old group.

Risk Factors of Urrets-Zavalia Syndrome after Penetrating Keratoplasty.

Objective: The objective of this study was to analyze the clinical features and risk factors of Urrets-Zavalia syndrome (UZS) after penetrating keratoplasty (PKP). Methods: The medical records of 152 patients who underwent PKP at the Department of Ophthalmology, Tongji Hospital, between January 2014 and December 2016 were retrospectively reviewed. UZS was diagnosed based on pre- and post-operative pupillary findings. The relationships among the primary disease, postoperative intraocular pressure (IOP), and the incidence of UZS were statistically analyzed. The pupillary changes during the follow-up period were studied. Results: Among the 152 included patients, 23 were diagnosed with UZS, with an incidence of 15.13%. The primary diseases of the UZS patients were keratoconus (eight cases, 34.78%), viral keratitis (six cases, 26.08%), leukoma (four cases, 17.39%), fungal corneal ulcer (two cases, 8.70%), corneal endothelial decompensation (two cases, 8.70%), and corneal degeneration (one case, 4.35%). The incidence of UZS in keratoconus patients was higher than that in patients with fungal corneal ulcer (42.11% versus 6.25%, p = 0.003); In addition, the transient postoperative high IOP was not significantly related to the incidence of UZS in keratoconus patients in our study (p = 0.319). Twenty-one patients with UZS were followed up for >6 months, seven of whom (33.33%) recovered spontaneously (within the range of 48 days to 1.5 years). Conclusion: In our study, the incidence of UZS after PKP was 15.13%, and 33.33% of these patients recovered spontaneously. UZS may be more likely to occur in patients with keratoconus. Postoperative transient high IOP may increase the incidence of UZS after PKP for keratoconus.

Enhancement of Boiling with Scalable Sandblasted Surfaces.

Surface engineering has been leveraged by researchers to enhance boiling heat transfer performance, with benefits ranging from improved thermal management to more efficient power generation. While engineered surfaces fabricated using cleanroom processes have shown promising boiling results, scalable methods for surface engineering are still limited despite most real-world industry-scale applications involving large boiling areas. In this work, we investigate the use of sandblasting as a scalable surface engineering technique for the enhancement of pool boiling heat transfer. We vary the size of an abrasive Al2O3 sandblasting medium (25, 50, 100, and 150 µm) and quantify its effects on silicon surface conditions and boiling characteristics. The surface morphology and capillary wicking performance are characterized by optical profilometry and capillary rise tests, respectively. Pool boiling results and surface characterization reveal that surface roughness and volumetric wicking rate increase with the abrasive size, which results in improvements in the critical heat flux and the heat transfer coefficient of up to 192.6 and 434.3% compared to a smooth silicon surface, respectively. The significant enhancement achieved with sandblasted surfaces indicates that sandblasting is a promising option for improving boiling performance in industry-scale applications.

Limbal Niche Cells and Three-Dimensional Matrigel-Induced Dedifferentiation of Mature Corneal Epithelial Cells.

Purpose: To investigate the phenotypic changes of mature corneal epithelial cells (MCECs) that cocultured with limbal niche cells (LNCs) in three-dimensional Matrigel (3D Matrigel) in vitro. Methods: MCECs were isolated from central corneas, and limbal epithelial progenitor cells (LEPCs) were isolated from limbal segments with Dispase II. LNCs were isolated and cultured from limbal niche using the collagenase A digestion method and identified with PCK/VIM/CD90/CD105/SCF/PDGFRß. MCECs were cultured on 3D Matrigel (50%, v/v) with or without LNCs for 10 days. Expression of CK12 and p63α and clone formation test were used to compare the progenitor phenotypic changes for MCECs before and after induction using LEPCs as control. Results: Homogeneous LNCs were isolated and identified as spindle shape and adherent to a plastic surface coated with 5% Matrigel. Double immunostaining of the fourth-passage LNCs was uniformly PCK-/VIM+/CD90+/CD105+/SCF+/PDGFRß+. Reverse transcription and quantitative real-time polymerase chain reaction (RT-qPCR) revealed the decrease of PCK expression from the second passage and elevation of Vim, CD90, CD105, SCF, and PDGFRß transcripts from the third passage, and the transcription level of Vim, CD90, CD105, SCF, and PDGFRß was elevated statistically in the fourth passage compared to the first passage (P < 0.01). Both immunofluorescence (IF) staining for cross section and cytospin cells demonstrated that MCECs expressed higher CK12 while lower p63α than LEPCs (P < 0.01). Sphere growth formation was noticed as early as 24 hours in the MCEC + LNC group, 48 hours in the LEPC group, and 72 hours in the MCEC group. The diameters of the spheres were the biggest in the MCEC + LNC group (182.24 ± 57.91 µm), smaller in the LEPC group (125.71 ± 41.20 µm), and smallest in the MCEC group (109.39 ± 34.85 µm) by the end of the 10-day culture (P < 0.01). Double immunostaining with CK12/p63α showed that cells in the sphere formed from MCECs expressed CK12 but not p63α; in contrast, some cells in the MCEC + LNC group expressed CK12, but most of them expressed p63α. RT-qPCR revealed a significant reduction of CK12 transcript but elevation of p63α, Oct4, Nanog, Sox2, and SSEA4 (P < 0.05). Holoclone composed of cubic epithelial cells could be generated in the MCEC + LNC group but not in the other two groups. Conclusions: The data shows that human MCEC cell phenotype could be induced to the dedifferentiation stage when cocultured with LNCs in 3D Matrigel that simulated the microenvironment of limbal stem cells in vitro.

Role of IL-36γ/IL-36R Signaling in Corneal Innate Defense Against Candida albicans Keratitis.

Purpose: Interleukin (IL)-36 cytokines have been shown to play either beneficial or detrimental roles in the infection of mucosal tissues in a pathogen-dependent manner, but their involvement in fungal keratitis remains elusive. We herein investigated their expression and function in mediating corneal innate immunity against Candida albicans infection. Methods: Gene expression in mouse corneas with or without C. albicans infection was determined by regular RT- and real-time (q)-PCR, Western blot analysis, ELISA or proteome profile assay. The severity of C. albicans keratitis was assessed using clinical scoring, bacterial counting, and myeloperoxidase (MPO) activity as an indicator of neutrophil infiltration. IL36R knockout mice and IL-33-specific siRNA were used to assess the involvement IL-33 signaling in C. albicans-infected corneas. B6 CD11c-DTR mice and clodronate liposomes were used to define the involvement of dendritic cells (DCs) and macrophages in IL-36R signaling and C. albicans keratitis, respectively. Results: IL-36γ were up-regulated in C57BL6 mouse corneas in response to C. albicans infection. IL-36 receptor-deficient mice display increased severity of keratitis, with a higher fungal load, MPO, and IL-1ß levels, and lower soluble sIL-1Ra and calprotectin levels. Exogenous IL-36γ prevented fungal keratitis pathogenesis with lower fungal load and MPO activity, higher expression of sIL-1Ra and calprotectin, and lower expression of IL-1ß, at mRNA or protein levels. Protein array analysis revealed that the expression of IL-33 and REG3G were related to IL-36/IL36R signaling, and siRNA downregulation of IL-33 increased the severity of C. albicans keratitis. Depletion of dendritic cells or macrophages resulted in severe C. albicans keratitis and yet exhibited minimal effects on exogenous IL-36γ-induced protection against C. albicans infection in B6 mouse corneas. Conclusions: IL-36/IL36R signaling plays a protective role in fungal keratitis by promoting AMP expression and by suppressing fungal infection-induced expression of proinflammatory cytokines in a dendritic cell- and macrophage-independent manner.