Guidelines for the application of artificial intelligence in the diagnosis of anterior segment diseases (2023).

The landscape of ophthalmology has observed monumental shifts with the advent of artificial intelligence (AI) technologies. This article is devoted to elaborating on the nuanced application of AI in the diagnostic realm of anterior segment eye diseases, an area ripe with potential yet complex in its imaging characteristics. Historically, AI's entrenchment in ophthalmology was predominantly rooted in the posterior segment. However, the evolution of machine learning paradigms, particularly with the advent of deep learning methodologies, has reframed the focus. When combined with the exponential surge in available electronic image data pertaining to the anterior segment, AI's role in diagnosing corneal, conjunctival, lens, and eyelid pathologies has been solidified and has emerged from the realm of theoretical to practical. In light of this transformative potential, collaborations between the Ophthalmic Imaging and Intelligent Medicine Subcommittee of the China Medical Education Association and the Ophthalmology Committee of the International Translational Medicine Association have been instrumental. These eminent bodies mobilized a consortium of experts to dissect and assimilate advancements from both national and international quarters. Their mandate was not limited to AI's application in anterior segment pathologies like the cornea, conjunctiva, lens, and eyelids, but also ventured into deciphering the existing impediments and envisioning future trajectories. After iterative deliberations, the consensus synthesized herein serves as a touchstone, assisting ophthalmologists in optimally integrating AI into their diagnostic decisions and bolstering clinical research. Through this guideline, we aspire to offer a comprehensive framework, ensuring that clinical decisions are not merely informed but transformed by AI. By building upon existing literature yet maintaining the highest standards of originality, this document stands as a testament to both innovation and academic integrity, in line with the ethos of renowned journals such as Ophthalmology.

Effect of palmitoylethanolamide on degeneration of a human-derived retinal pigment epithelial cell induced by all-trans retinal.

AIM: To study the effect of palmitoylethanolamide (PEA) on apoptosis of retinal pigment epithelial (RPE) cells induced by all-trans retinal (atRAL) and to explore the possible molecular mechanism. METHODS: CellTiter 96® Aqueous One Solution Cell Proliferation Assay (MTS) was used to detect the effect of PEA on human-derived retinal epithelial cells (ARPE-19) viability induced by atRAL. A Leica DMi8 inverted microscope was used to observe cell morphology. Reactive oxygen species (ROS) production was evaluated with 2',7'-dichlorodihydrof-luorescein diacetate (H2DCFDA) staining and fluorescence microscopy. Expression of c-Jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK), c-Jun, phosphorylated c-Jun (p-c-Jun), Bak, cleaved caspase-3, C/EBP homologous protein (CHOP), and binding (Bip) protein levels were tested by Western blot. Abca4 -/- Rdh8 -/- mice, mouse models of atRAL clearance defects which displays some symbolic characteristics of dry age-related macular degeneration (AMD) and Stargardt disease (STGD1). In the animal models, PEA was injected intraperitoneally. The full-field electroretinogram was used to detect visual function under scotopic conditions traced from mice. Optical coherence tomography showed reconstitution or thickening of the retinal pigment epithelium layer. Effect of PEA on fundus injury induced by light in Abca4-/-Rdh8-/- mice was observed by fundus photography. RESULTS: PEA ameliorated ARPE-19 cells apoptosis and inhibited ROS (including mitochondrial ROS) production induced by atRAL. PEA improved the retinal functional, prohibited both RPE and photoreceptor from death, ameliorates light-induced fundus impairment in Abca4 -/- Rdh8 -/- mice. In vitro and in vivo, PEA inhibited JNK, p-JNK, c-Jun, p-c-Jun, Bak, cleaved caspase-3, CHOP, and Bip protein levels induced by all-trans retinal in ARPE-19 cells. CONCLUSION: PEA has effect on treating RPE cells apoptosis in retinopathy caused by atRAL accumulation. PEA is a potential treatment strategy for dry AMD and STGD1. The molecular mechanism is affecting the ROS-JNK-CHOP signaling pathway partly.

Tear dynamics testing and quantitative proteomics analysis in patients with chronic renal failure.

Ocular surface changes may develop in patients with chronic renal failure (CRF) undergoing hemodialysis. In recent years, an association of CRF with dry eye syndrome has been emphasized. However, tear proteomics of CRF patients has not been analyzed. Here, we performed systematic profiling of the tear film proteins in CRF patients through use of isobaric tags for relative and absolute quantitative (iTRAQ) MS/MS, aiming to identify associations between dry eye symptoms and expression of tear proteomic changes in patients with CRF undergoing hemodialysis. Twenty CRF patients and ten healthy subjects underwent a series of ophthalmic examinations. Tear samples from the participants were analyzed by iTRAQ approach. A total of 1139 tear proteins were screened, and 212 differentially expressed proteins were identified. The pattern changes included 77 whose expression levels were upregulated (fold increase >1.2) whereas 135 others that were downregulated (fold decrease <1/1.2). Bioinformatics analysis showed that these proteins were significantly enriched in lipid metabolism, inflammatory, and immune response pathways. Furthermore, APOA1, APOA4, APOB, APOE, S100A8, S100A9, S100A4, HSP90B and other molecules were significantly changed. Our study elucidated the characteristics of tear dynamics and protein markers in CRF patients undergoing hemodialysis. Significance: Despite the association of chronic renal failure (CRF) with dry eye disease, there are no reports describing potentially important differentially expressed tear proteins in CRF patients undergoing hemodialysis. It is still a challenge to obtain a comprehensive description of the pathogenesis of dry eye in CRF patients which hinders establishing a patient specific therapeutic scheme. Our study is the first iTRAQ proteomics analysis of the tears of patients with CRF, which reveals the changes in the protein expression profile in CRF patients afflicted with dry eye disease. The identity was verified of some relevant differentially expressed proteins, and they may be candidate diagnostic markers of dry eye disease in patients with CRF. These tear film protein constituents found in hemodialysis patients can be of important clinical significance in treating this condition. SIGNIFICANCE: Despite the association of chronic renal failure (CRF) with dry eye disease, there are no reports describing potentially important differentially expressed tear proteins in CRF patients undergoing hemodialysis. It is still a challenge to obtain a comprehensive description of the pathogenesis of dry eye in CRF patients which hinders establishing a patient specific therapeutic scheme. Our study is the first iTRAQ proteomics analysis of the tears of patients with CRF, which reveals the changes in the protein expression profile in CRF patients afflicted with dry eye disease. The identity was verified of some relevant differentially expressed proteins, and they may be candidate diagnostic markers of dry eye disease in patients with CRF. These tear film protein constituents found in hemodialysis patients can be of important clinical significance in treating this condition.

Comprehensive strategy improves the genetic diagnosis of different polycystic kidney diseases.

Polycystic kidney disease (PKD) is known to occur in three main forms, namely autosomal dominant PKD (ADPKD), autosomal recessive PKD (ARPKD) and syndromic PKD (SPKD), based on the clinical manifestations and genetic causes, which are diagnosable from the embryo stage to the later stages of life. Selection of the genetic test for the individuals with diagnostic imaging reports of cystic kidneys without a family history of the disease continues to be a challenge in clinical practice. With the objective of maintaining a limit on the time and medical cost of the procedure, a practical strategy for genotyping and targeted validation to resolve cystogene variations was developed in our clinical laboratory, which combined the techniques of whole-exome sequencing (WES), Long-range PCR (LR-PCR), Sanger sequencing and multiplex ligation-dependent probe amplification (MLPA) to work in a stepwise approach. In this context, twenty-six families with renal polycystic disorders were enrolled in the present study. Thirty-two variants involving four ciliary genes (PKD1, PKHD1, TMEM67 and TMEM107) were identified and verified in 23 families (88.5%, 23/26), which expanded the variant spectrum by 16 novel variants. Pathogenic variations in five foetuses of six families diagnosed with PKD were identified using prenatal ultrasound imaging. Constitutional biallelic and digenic variations constituted the pathogenic patterns in these foetuses. The preliminary clinical data highlighted that the WES + LR PCR-based workflow followed in the present study is efficient in detecting divergent variations in PKD. The biallelic and digenic mutations were revealed as the main pathogenic patterns in the foetuses with PKD.

Novel Biallelic DNAH1 Variations Cause Multiple Morphological Abnormalities of the Sperm Flagella.

Sperm motility is vital to human reproduction, and malformed sperm flagella can cause male infertility. Individuals with multiple morphological abnormalities of the flagella mostly have absent, short, coiled, bent, and/or irregular-caliber flagella. In this study, a patient with male infertility underwent a physical examination along with his wife. Genetic testing was performed by whole-exome sequencing of the couple, and Sanger sequencing was performed for validation. Novel biallelic variations in the DNAH1: (NM_015512.4) gene consisting of c.1336G>C (p.E446Q) and c.2912G>A (p.R971H) were identified. In silico structural analysis revealed that the amino acid residues affected by the variation were evolutionarily conserved, and the variant p.R971H influenced the stability of the DNAH1 protein. Morphological studies of the patient's sperm showed defects in its flagella. Results of Papanicolaou staining and scanning electron microscopy demonstrated coiled and short flagella with multiple anomalies. Transmission electron microscopy of the sperm flagella showed that the inner dynein arm and radial spoke were absent, and the dense fiber and microtubule doublets were displaced. Quantitative PCR of the mRNA of the patient's sperm showed that the expression of DNALI1 was dramatically reduced. Collectively, these findings elucidated the genetic cause of the family's infertility and provided insight into the functioning of the DNAH1 gene.

Netrin-1 promotes epithelium repair in corneal injury.

AIM: To explore netrin-1 functions on corneal epithelium in vitro and in vivo. METHODS: In vitro the human corneal epithelial (HCE) cells were treated with serum free DMEM-F12 basic media containing 0, 50, 100, 200, 300, 500, 800, and 1000 ng/mL of netrin-1, respectively. The cells viability was detected by cell counting kit-8 (CCK-8). The wound-healing assay was applied to assess the migration proficiency of HCE cells. Flow cytometry was used to analyze the cell-cycle distribution and apoptosis. In vivo, normal c57 (6wk) mice were demarcated with a trephine in the middle of the cornea to produce a 3-mm circular wound. Mice corneas were inflicted no epithelium with a 3-mm wound displayed, but remained the limbal epithelium intact. A blunt scalpel blade was used to remove the corneal epithelian cells, followed by topical netrin-1 application (200 ng/mL), and the group treated by PBS as control. The treated group was injected netrin-1 into the normal c57 mice inferior subconjunctival 4h before trauma. Mouse corneal inflammation and neovascularization were observed under slit lamp microscope. The apoptosis of corneal cells was determined by TUNEL staining. RESLUTS: A concentration of 200 ng/mL netrin-1 enhanced 25% of the HCE viability. The relative migration rates were 76.3% and 100% in control and netrin-1 treated group after cultured 72h. Treated with netrin-1 (200 ng/mL) decreased the apoptosis of HCE cells, as well as decreased their percentage from 19.3%±0.57% to 12.7%±0.42% of the total. The remaining wound area was 1.22 mm2 in control group but 0.22 mm2 in the netrin-1 treated group. Exogenous Netrin-1 inhibits apoptosis of corneal epithelial cells of c57 mice. TUNEL-positive cells at the epithelial layer of the corneas of the control and netrin-1 treated c57 mice at 24h after wounding were 43.3% and 16.7% respectively. CONCLUSION: Netrin-1 can reduce HCE apoptosis as well as promote its proliferation and migration. Topical application of netrin-1 promotes the injuryed cornea epithelial wound repair and inhibits apoptosis of corneal epithelial cells. These findings may offer potential therapies to repair the defects of corneal epithelial based on netrin-1.

The prognosis and effects of local treatment strategies for orbital embryonal rhabdomyosarcoma: a population-based study.

INTRODUCTION: Orbital embryonal rhabdomyosarcoma is a rare childhood malignancy with a good prognosis, but the optimal treatment remains unclear. Using a population-based cancer registry, we assessed the prognoses and survival outcomes of patients with orbital embryonal rhabdomyosarcoma according to the local treatment strategy. PATIENTS AND METHODS: Patients diagnosed with orbital embryonal rhabdomyosarcoma between 1988 and 2012 as part of the Surveillance Epidemiology and End Results program were included. Univariate and multivariate Cox regression analyses were performed to determine the prognostic factors associated with cause-specific survival (CSS) and overall survival (OS). RESULTS: In total, 102 patients were included; their median age was 6 years, 78.4% were white, and 56.9% were male. The median tumor size was 30 mm. Of 20 patients with an available histologic grade, the tumors of 90% were poorly differentiated/undifferentiated. Of 92 patients with available surgical and radiotherapy (RT) statuses, 50 (54.3%), 36 (39.1%), and 6 (6.5%) received surgery and RT, primary RT, and primary surgery, respectively. Ninety-five patients (93.1%) received chemotherapy. The 5- and 10-year CSSs of the entire cohort were 94.3% and 92.2%, respectively. The 5- and 10-year OSs were 93.3% and 91.3%, respectively. In 95 patients who were followed up for at least 12 months, there were no significant prognostic factors related to CSS and OS. Furthermore, the local treatment strategy did not significantly affect CSS (P=0.29) or OS (P=0.468). CONCLUSION: There is no local treatment of choice for orbital embryonal rhabdomyosarcoma in terms of survival. However, RT is a reasonable alternative treatment to surgery.

Therapeutic effects of topical netrin-4 in a corneal acute inflammatory model.

AIM: To evaluate the therapeutic effect of netrin-4 on the early acute phase of inflammation in the alkali-burned eye. METHODS: Eye drops containing netrin-4 or phosphate buffered saline (PBS) were administered to a alkali-burn-induced corneal acute inflammatory model four times daily. The clinical evaluations, including fluorescein staining and inflammatory index, were performed on day 1, 4 and 7 using slit lamp microscopy. Global specimens were collected on day 7 and processed for immunofluorescent staining. The levels of inflammatory mediators in the corneas were determined by real-time polymerase chain reaction (PCR). RESULTS: Exogenous netrin-4 administered on rat ocular surfaces showed more improvements in decreasing fluorescein staining on day 4 and 7, and resolved alkali burn-induced corneal inflammation index on day 7 (P<0.01). The levels of IL-1ß, IL-6, intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-1 (MIP-1) in corneas were decreased in netrin-4-treated groups (P<0.05). In addition, netrin-4 significantly reduced the expression of leukocyte common antigen 45 (CD45) in the alkali-burn cornea (P<0.001). CONCLUSION: Topical netrin-4 accelerated wound healing and reduced the inflammation on alkali-burn rat model, suggesting a potential as an anti-inflammatory agent in the clinical to treat the acute inflammation.

Differential expression and function of survivin during the progress of pterygium.

PURPOSE: To investigate the expression pattern and function of survivin in the development of pterygium. METHODS: Primary pterygia at quiescent or advanced clinical stage and normal human conjunctival tissues were used in this study. Pterygium epithelial cells (PECs) were cultured in keratinocyte serum-free defined medium and harvested at different growth stages. Tissue sections and cultured cells were detected with survivin, phosphorylated survivin (Thr43), p63, p57, and p21 on protein, and/or mRNA level. Cell Counting Kit (CCK)-8 assay was performed to measure proliferation status of primary cultured PECs. Small interfering (si) RNA specific for survivin was transfected on PECs at subconfluence stage. RESULTS: Survivin was highly expressed in all pterygium tissues, but not in normal human conjunctiva, at mRNA and protein levels. It was mainly present in the epithelial cytoplasm of pterygium at quiescent stage, while present in the nucleus of pterygium at advanced stage. Phosphorylated survivin was upregulated in pterygium at advanced stage. Pterygium epithelial cells cultured under subconfluence stage showed higher expression of survivin and p63, but lower expression of p57 and p21, compared with PECs reached confluence. Both total and phosphorylated survivin was mainly expressed in the nuclei of PECs under subconfluence, and there was cytoplasmic translocation of survivin when PECs reached confluence. The knockdown of survivin by siRNA inhibited proliferation of PECs, accompanied by downregulation of p63, and upregulation of p57 and p21. CONCLUSIONS: Higher subcellular expression and phosphorylation of survivin may play roles in the development of pterygium. Survivin could be targeted for the treatment of pterygium.

Serine Proteinase Inhibitor SERPINA3K Suppresses Corneal Neovascularization via Inhibiting Wnt Signaling and VEGF.

Purpose: To evaluate the anti-neovascularization effects and investigate the possible mechanisms of SERPINA3K, a member of serine proteinase inhibitor family, using a specific rat model of suture-induced corneal neovascularization. Methods: A rat corneal suture model was set up and SERPINA3K was topically administered three times daily for 7 days. The clinical indications were evaluated on day 2, 5 and 7, including area of neovascularization and inflammation index. The eyeballs were collected after day 7 and the following examinations were performed: histological investigation, immunostaining, western blot and quantitative real-time polymerase chain reaction (PCR) assay. Wnt3a, a Wnt pathway ligand, was added to cultured Human Umbilical Vein Endothelial Cells (HUVEC), followed by detecting cell migration and western blot. Meanwhile, an in vitro VEGF165-stimulated HUVEC model was applied and the following measurements were conducted: cell proliferation, cell migration and tube formation. Results: SERPINA3K significantly suppressed corneal neovascularization and inhibited corneal inflammation. SERPINA3K downregulated the levels of ß-catenin, non-pi-ß-catenin and transcription factor 4 (TCF4), but upregulated the level of pi-ß-catenin of the corneas induced by suture. SERPINA3K also decreased the gene expression and protein level of VEGF. Meanwhile, induction of Wnt3a increased the cell migration, activated the Wnt signaling and upregulated VEGF in cultured HUVEC, which were antagonized by SERPINA3K. In addition, SERPINA3K significantly inhibited VEGF165-induced cell proliferation and migration of HUVEC, SERPINA3K also specifically suppressed the VEGF165-induced tube formation of HUVEC. Conclusions: SERPINA3K has therapeutic potential for corneal neovascularization. The underlying mechanism may be through inhibiting Wnt signaling pathway and VEGF.

Therapeutic effects of topical doxycycline in a benzalkonium chloride-induced mouse dry eye model.

PURPOSE: We investigated the therapeutic effects and underlying mechanisms of topical doxycycline in a benzalkonium chloride (BAC)-induced mouse dry eye model. METHODS: Eye drops containing 0.025%, 0.1% doxycycline or solvent were administered to a BAC-induced dry eye model four times daily. The clinical evaluations, including tear break-up time (BUT), fluorescein staining, inflammatory index, and tear volume, were performed on days 0, 1, 4, 7, and 10. Global specimens were collected on day 10 and processed for immunofluorescent staining, TUNEL, and periodic acid-Schiff assay. The levels of inflammatory mediators in the corneas were determined by real-time PCR. The total and phosphorylated nuclear factor-κB (NF-κB) were detected by Western blot. RESULTS: Both 0.025% and 0.1% doxycycline treatments resulted in increased BUT, lower fluorescein staining scores, and inflammatory index on days 4, 7, and 10, while no significant change in tear volume was observed. The 0.1% doxycycline-treated group showed more improvements in decreasing fluorescein staining scores, increasing Ki-67-positive cells, and decreasing TUNEL- and keratin-10-positive cells than other groups. The mucin-filled goblet cells in conjunctivas were increased, and the expression of CD11b and levels of matrix metalloproteinase-9, IL-1ß, IL-6, TNF-α, macrophage inflammatory protein-2, and cytokine-induced neutrophil chemoattractant in corneas were decreased in both doxycycline-treated groups. In addition, doxycycline significantly reduced the phosphorylation of NF-κB activated in the BAC-treated corneas. CONCLUSIONS: Topical doxycycline showed clinical improvements and alleviated ocular surface inflammation on BAC-induced mouse dry eye, suggesting a potential as an anti-inflammatory agent in the clinical treatment of dry eye.

[Abnormal epithelial differentiation and tear film alteration in pterygium].

OBJECTIVE: To investigate the differentiation and proliferation of the conjunctival epithelium and tear film function in pterygia. METHODS: It was a retrospective study. Fifteen patients (15 eyes) who underwent excision for pterygium were enrolled in this study. Immunostaining for K10, K14, K19, MUC5AC, K16, Ki67 and P63 was performed on the pterygial epithelium and normal conjunctival epithelium. Schirmer I test was performed, and the tear film break-up time (BUT) was evaluated just prior to and 6 months after surgery. Multi-factor regression analysis was assessed to observe if there is a correlation between pterygial growth and tear film function. RESULTS: The average absorbency of K19 and MUC5AC immuno-staining all significantly changed (3727.86 ± 2544.73 vs. 25 528.00 ± 12 901.06, 2080.48 ± 2340.17 vs. 7182.51 ± 3069.20, t = 9.261,3.538, P < 0.05), and increased in K10 and K14 in patients with pterygia compared with normal conjunctivae keratin (2017.51 ± 2114.3 vs. 0, 6027.5 ± 1058.32 vs. 2123.28 ± 1249.09, t = -6.151, P < 0.05). Furthermore, pterygial epithelium showed activated proliferation, evidenced by significantly up-regulated expression of K16, P63 and Ki67 compared to normal control. The Schirmer I test did not indicate any significant differences pre- and post-operatively. However, the BUT was significantly prolonged 1 month post surgery compared to pre-surgery (t = -4.222, P < 0.05). CONCLUSION: This study indicates that abnormal epithelial differentiation and proliferation are present in pterygium , which is characterized by squamous metaplasia, accompanied with instability of tear film and normal basic tear secretion.

[The clinical efficiency of calf blood extract gel on moderate to severe dry eye induced by chronic graft versus host diseases after bone marrow transplantation].

OBJECTIVE: To investigate the clinical efficiency of calf blood extract gel on dry eye induced by chronic graft versus host diseases after bone marrow transplantation. METHODS: It was a two-stage cross-over design double-blind controlled study. Twelve patients (twenty-four eyes) diagnosis dry eye induced by chronic graft versus diseases in Ocular Surface Out-patient Clinic of Xiamen University Affiliated Xiamen Eye Center 2009 from 2010 to were divide into two groups: group A accepted the treatment of autologous serum in the first stage and group B accepted the treatment of calf blood extract gel, after one month of elution, group A accepted calf blood extract gel and group B accepted autologous serum. The signs and symptoms with different therapies were recorded at the time of pre and post therapies, which were analyzed by Wilcoxon analysis and two-stage cross-over analysis. RESULTS: Ocular dry eye symptoms including visual tiredness, dry and unsmooth sensation, foreign body sensation, photophobia, pain, redness and visual acuity had been improved in both autologous serum therapy (U = 22.5, 43.2, 27.0, 17.4, 21.5, 38.5, 23.0, P < 0.05) and calf blood extract gel therapy (U = 333.0, 24.5, 29.0, 40.5, 26.0, 36.0, 51.0, P < 0.05) after two-week treatment. Corneal FL had significantly been improved (2.00 ± 1.00, 3.00 ± 1.50) (Group A U = 273.0, Group B U = 135.0, P < 0.01). Ocular dry eye signs and symptoms including visual tiredness, dry and unsmooth sensation, burning sensation, photophobia, pain, tearing, redness, visual acuity, corneal FL, TFBUT and S It hadn't significantly improved between two kinds of therapies (F = 1.45, 2.43, 2.14, 1.91, 1.63, 0.51, 1.19, 0.68, 2.75, 0.77, 1.23, P > 0.05) or between two kinds of offering drug orders (F = 3.17, 2.62, 0.91, 1.42, 0.89, 2.17, 0.95, 1.54, 3.21, 6.72, 1.37, P > 0.05) in the two-stage cross-over design. Only foreign body sensation had significantly statistical difference between two kinds of drug (F = 11.38, P < 0.05), while without significant statistical difference between two kinds of offering drug orders (F = 2.62, P > 0.05). CONCLUSION: Calf blood extract gel can be consider as a alternative for the treatment of dry eye induced by cGVHDs, because of its functions on releasing ocular dry eye symptoms and promoting corneal epithelial cells repair.

[Emphasizing the clinical investigation of dry eye].

In the past twenty years, with the rapid development of dry eye research in China, significant progresses have been achieved and there has been a remarkable improvement in diagnostic rate and clinical outcomes. Publication and popularization of the clinical diagnosis and treatment recommendations for dry eye from consensus of expert advice provides criteria for clinical work of dry eye. There has still been a significant gap between our research and world's advanced level in dry eye, therefore clinical investigations of high level will be intensively promoted to reveal the epidemiologic and clinical characteristics of dry eye populations in China, and establish the diagnostic criteria for Chinese patients, develop the clinical research on dry eye drugs and home-made clinical devices. Based on evidence-based research evidences, criteria for diagnosis and treatment of dry eye will be established and the clinical investigation of dry eye will be developed to reach the world's advanced level.

[Abnormal of tear lipid layer and recent advances in clinical study of dry eye].

Dry eye is a common disease in the ophthalmological clinic, which is related to the dysfunction of tear film. The tear film is composed of lipid layer, aqueous layer and mucin layer (or lipid layer, aqueous/mucin layer). The lipid of the outmost layer derived from Meibomian gland and distributed on the tear film after blinking can decrease the evaporation and stabilize the tear film. The thickness, quality, and distribution of lipid layer are impaired in many dry eye patients, hence restoring the physiological function of lipid layer may be crucial for the treatment of this kind of dry eye. The lipid artificial tears manifest great effects on increasing lipid layer thickness, stabilizing tear film, improving Meibomian gland dysfunction, and promoting tear film distribution.

[Therapeutic effects of Pyranoprofen on the mouse dry eye induced by topical medication of Benzalkonium Chloride].

OBJECTIVE: Randomized controlled experimental study to investigate the therapeutic effect and the possible mechanism of Pranoprofen on the recovery of dry eye induced by topical medication of Benzalkonium Chloride (BAC) in mouse. METHODS: It was an experimental study. Seventy BALB/c mice were treated with topical administration of 0.25% BAC to establish the dry eye condition. Based on the consistency of break-up time of tear-film (BUT), corneal fluorescein staining scores and inflammation index, the eyes were re-selected and randomly divided into four groups on day (D) 21 after the BAC treatment. Group A was set up as blank control, while group B, C and D were treated respectively with 0.1% sodium hyaluronate eye drops, 0.1% fluorometholone eye drops plus 0.1% sodium hyaluronate eye drops, 0.1% pyranoprofen eye drops plus 0.1% sodium hyaluronate eye drops. BUTs, tear volumes, corneal fluorescein staining scores and inflammation index were evaluated in each group on D0, 1, 3 and 5 after the therapeutic treatment. Global specimens were collected on D6. Sections were stained with hematoxylin and eosin (HE) or by periodic acid-schiff (PAS) assay, and labeled with cytokeratin 10 (K10) antibody. The expression of tumor necrosis factor-α (TNF-α) in the cornea and conjunctiva was quantified by western blot. RESULTS: 72 eyes were included in the sequential experiment, 18 eyes for each group. On D0, 1 and 3, no clinical differences were observed among the groups. On D5, the BUT was (2.933 ± 0.320), (2.900 ± 0.280), (3.464 ± 0.498) and (3.643 ± 0.413) s in group A, B, C and D respectively; the BUTs in group C and D were significant longer than those of group A and B (F = 13.774, P = 0.000). The corneal fluorescein staining score was (11.640 ± 1.008), (11.790 ± 1.188), (10.330 ± 1.371) and (10.270 ± 1.104)s in group A, B, C and D respectively; the scores in group C and D were significant lower than those of group A and B (F = 6.145, P = 0.001). The corneal inflammatory index was (0.232 ± 0.059), (0.229 ± 0.078), (0.151 ± 0.055) and (0.154 ± 0.056) in group A, B, C and D respectively; the index in group C and D were significant lower than those of group A and B (F = 6.703, P = 0.001). No significant difference was found in tear volume among groups. No significant difference was found between Pyranoprofen and Fluorometholone treatment in BUT, corneal fluorescein score or inflammatory index. Corneal morphology showed the feature of thicker corneal epithelial layer in group A and uniformity in group C and D. PAS assay revealed similar goblet cell numbers in group C and D, but less goblet cells in group A and B. Cytokeratin 10 was almost negatively expressed in Pranoprofen or Fluorometholone treated groups, and remained positive in the corneal epithelium with other treatments. The level of TNF-α in the cornea was down-regulated in Pranoprofen or Fluorometholone treated groups. CONCLUSIONS: Pranoprofen or Fluorometholone combined with sodium hyaluronate treatment presented similar therapeutic effects on BAC-induced mouse dry eye, with the more stable tear film, the better regularity of epithelium recovery, the down-regulation of inflammatory TNF-α, the increased number of goblet cells, and the elimination of squamous metaplasia, when compared with the treatment of sodium hyaluronate eye drops only. Our results showed the great potentialities of Pranoprofen in the clinical treatment of ocular surface inflammation in the mild and severity dry eye.

Menin expression is regulated by transforming growth factor beta signaling in leukemia cells.

BACKGROUND: Menin is a ubiquitously expressed protein encoded by the multiple endocrine neoplasia type 1 (MEN1) gene. Besides its importance in endocrine organs, menin has been shown to interact with the mixed lineage leukemia (MLL) protein, a histone H3 lysine 4 methyltransferase, and plays a critical role in hematopoiesis and leukemogenesis. Previous studies have shown that menin promotes transforming growth factor beta (TGF-ß) signaling in endocrine cells. However, little is known regarding the impact of TGF-ß pathway on menin in hematopoietic system. Here, with leukemia cell lines generated from conditional MEN1 or TGF-ß receptor (TßRII) knockout mouse models, we investigated the possible cross-talk of these two pathways in leukemia cells. METHODS: MEN1 or TßRII conditional knockout mice were bred and the bone marrow cells were transduced with retroviruses expressing oncogeneic MLL-AF9 (a mixed lineage leukemia fusion protein) to generate two leukemia cell lines. Cell proliferation assays were performed to investigate the effect of TGF-ß treatment on MLL-AF9 transformed leukemia cells with/without MEN1 or TßRII excision. Menin protein was detected with Western blotting and mRNA levels of cell proliferation-related genes Cyclin A(2) and Cyclin E(2) were examined with real-time RT-PCR for each treated sample. In vivo effect of TGF-ß signal on menin expression was also investigated in mouse liver tissue after TßRII excision. RESULTS: TGF-ß not only inhibited the proliferation of wild type MLL-AF9 transformed mouse bone marrow cells, but also up-regulated menin expression in these cells. Moreover, TGF-ß failed to further inhibit the proliferation of Men1-null cells as compared to Men1-expressing control cells. Furthermore, excision of TßRII, a vital component in TGF-ß signaling pathway, down-regulated menin expression in MLL-AF9 transformed mouse bone marrow cells. In vivo data also confirmed that menin expression was decreased in liver samples of conditional TßRII knockout mice after TßRII excision. CONCLUSION: These results provided the first piece of evidence of cross-talk between menin and TGF-ß signaling pathways in regulating proliferation of leukemia cells, suggesting that manipulating the cross-talk of the two pathways may lead to a novel therapy for leukemia.

[The morphology and thickness of cornea in patients with Marfan syndrome].

OBJECTIVE: To search for the characteristics of MFS in corneal morphology and thickness. METHODS: Twenty-four patients (48 eyes) with MFS and 24 healthy age- and gender-matched volunteers (48 eyes) were recruited in this clinical prospective, and comparative series study. Firstly, biomicroscopic examination and Type-A ultrasonometry was conducted to search for ectopia lentis and axis length. Secondly, the corneal morphologic parameter [including the height of anterior and posterior surface, the centre corneal curvature, the mean astigmatism in the 3.0-mm central zone (Mean A), the mean simulated astigmatism (Sim A), the mean keratometry in the 3.0-mm central zone (Mean K), the mean simulated keratometry (Sim K), the 3.0-mm zone irregularity (3.0ZI), the 5.0-mm zone irregularity (5.0ZI), corneal thickness index (CTI)] and thickness (at the central location and at eight midperipheral locations) were obtained by the the autorefractometer and the Orbscan II Z corneal topography. Last, the statistics method including Crosstabs, One-way ANOVA, student-t test and discriminant analysis were applied and the correlations were established. RESULTS: There is no statistically significance between MFS group and control group in ages (38 ± 7) and (37 ± 8) years, gender (8/16) and (9/15), and axis length (23.12 ± 1.06) mm and (24.26 ± 2.96) mm (age χ(2) = 0.091, P = 0.763;gender t = 0.324, axis length t = 1.976, P > 0.05). Flat cornea ratio (66.7% and 12.5%) and topography of the oval (25.0% and 16.7%), irregular bow-shaped (41.7% and 37.5%) and irregular-shaped (12.5% and 8.3%) were increased significantly in patients with MFS. The corneal topography (MFS/control) showed that there are statistically significance in the thinnest thickness of cornea (489.8 ± 42.9)µm and (544.8 ± 25.7)µm, Mean K (40.60 ± 1.30) D and (42.80 ± 1.40) D, Sim K (40.50 ± 1.30) D and (42.80 ± 1.20) D, Sim A (1.08 ± 0.86)D and (0.91 ± 0.46) D, CTI 1.57 ± 0.24 and 1.21 ± 0.14, 3.0ZI (1.76 ± 0.96) D and (1.54 ± 0.82) D, and 5.0ZI (1.91 ± 1.26) D and (0.92 ± 0.68) D (thinnest thickness t = 6.996, Mean K t = 2.554, Sim K t = 3.326, Sim A t = 2.324, CTI t = 3.116, 3.0ZI t = 2.686, 5.0ZI t = 3.768, P < 0.05), while no statistically significance in the Mean A between the MFS (1.11 ± 0.89) D and control group (0.99 ± 0.49) D (Mean A t = 1.898, P = 0.08); except for temple inferior, the significant decrease of pachymetry (including the center and the seven midperipheral locations) appeared in the MFS group compared with the control group. CONCLUSION: The characteristic of MFS in corneal topography is that corneal axial refractive power descends and corneal thickness decreases.

[Focus on the challenge of basic research on corneal tissue engineering].

Great progress has been made on the corneal tissue engineering in the past two decades. Much knowledge has been gained on the seed cells, carrier material, and strategies of corneal tissue reconstruction. However, there are still great challenges regarding the basic research of corneal tissue engineering, such as selection of appropriate carrier and cells, optimization and standardization of the construction method, and evaluation of the clinic outcome. Future studies may address these questions and bring tissue engineered cornea into clinic application.

[Preliminary investigation on tear film alterations in latent herpes stromal keratitis].

OBJECTIVE: To investigate tear film alterations in patients with latent herpes stromal keratitis (HSK). METHODS: Prospective comparative case series study. Twenty-four patients with latent HSK in one eye and 28 age and gender matched healthy individuals were recruited. All subjects were evaluated by subjective symptoms of dry eye, tear film break-up time (BUT) and Schirmer I test (SIT). Laser in vivo confocal microscopic investigation was performed in 12 patients with severe tear film instability (BUT ≤ 5 s). Data distribution and homogeneity of variance was analyzed. Statistical comparisons of the mean values between different groups were performed using Kruskal-Wallis test, Mann-Whitney test or student t-test. RESULTS: Most of latent HSK patients (n = 22/24, 91.7%) had symptoms as dryness, burning sensation, redness and foreign body sensation. Both eyes of patients with latent HSK had hyposecretion (SIT, control eyes (16.2 ± 3.2) mm/5 min; affected eyes (10.4 ± 7.8) mm/5 min; lateral eyes (11.2 ± 8.8) mm/5 min; control and affected, U = 135.0, P < 0.001; control and lateral, U = 155.0, P = 0.001) and decreased tear film stability [BUT, control eyes (12.1 ± 0.7) s, affected eyes (4.3 ± 3.3) s, lateral eyes (9.2 ± 4.4) s; control and affected, U = 28.0, P < 0.001; control and lateral, U = 114.0, P < 0.001] as compared to control group (Kruskal-Wallis nonparametric test). The value of BUT showed significant difference between affected eyes and healthy eyes (U = 90.0, P < 0.001), whereas no difference of the value of SIT was found (U = 273.0, P = 0.757). Abnormal SIT (≤ 10 mm/min) and BUT (≤ 10 s) was presented in 14 (58.3%) and 23 (95.8%) affected eyes, as well as in 14 (58.3%) and 17 (70.8%) lateral eyes, respectively. Laser in vivo confocal microscopy investigation in 12 affected corneas with abnormal tear film showed morphological alterations as corneal epithelial metaplasia with polymorphism and enlarged cells, reflective nuclei, and decreased nucleus/cytoplasm ratio; decreased nerve density in subepithelial plexus and obvious branching and beading, which is similar to those changes caused by dry eye. CONCLUSIONS: Most of latent HSK patients had abnormal tear film. Dry eye related alterations could be found in affected corneas with abnormal tear film by in vivo confocal microscopy.