Thalidomide, a glutamate derivative with teratogenicity, possesses anti-inflammatory, immunomodulatory, and anti-angiogenic properties that enable its use in treating refractory diseases unresponsive to conventional therapies. Dry age-related macular degeneration (AMD), characterized by retinal pigment epithelium (RPE) degeneration and lacking effective therapies, represents a significant unmet medical need. Our findings demonstrated that thalidomide significantly restores mitochondrial function, alleviates G2/M phase cell cycle arrest, and suppresses sustained endoplasmic reticulum (ER) stress in oxidatively injured RPE cells. Mechanistically, these effects are coordinated through E2F2 activation, which subsequently regulates FBXO5 expression. Moreover, thalidomide was able to ameliorate oxidative stress-induced retinal structural disorders and RPE degeneration, and improve visual function in mice. In summary, this study elucidates that thalidomide synergistically regulates cell cycle progression and endoplasmic reticulum homeostasis through the E2F2-FBXO5 signaling pathway, providing a new drug candidate and therapeutic target for the prevention and treatment of dry AMD.

Simultaneously restricting bacterial proliferation, mitigating inflammatory reactions and excessive reactive oxygen species (ROS)-mediated corneal damage during sleep represents a crucial therapeutic strategy for managing bacterial keratitis (BK).

OBJECTIVE:To investigate the protective effects and molecular mechanisms of apigenin (API) on lipopolysaccharide (LPS)-induced corneal inflammation. METHODS:Immortalized human corneal epithelial cell line (HCECs) and primary human corneal epithelial cells (PHCECs) were used to establish LPS-induced inflammation models in vitro. IL-6 and IL-8 mRNA and protein levels were assessed by qRT-PCR and ELISA. Transcriptome sequencing and KEGG/GO enrichment analyses were performed to identify key pathways. Western blotting evaluated the activation of MAPK (ERK, JNK, P38) and NF-κB (P65, IκBα) pathway proteins, and immunofluorescence was used to examine P65 nuclear translocation. Furthermore, following the establishment of an inflammation model via intrastromal LPS injection in mice, API was administered to assess its impact on ocular inflammation and pro-inflammatory cytokine expression. RESULTS:API significantly suppressed the expression and secretion of key pro-inflammatory mediators in LPS-stimulated corneal epithelial cells including IL-6, IL-8, and COX2. Transcriptomic analysis confirmed the MAPK and NF-κB pathways as critical components in API'a anti-inflammatory action. API inhibited the phosphorylation of key proteins in the MAPK-JNK/ERK and NF-κB signaling pathways, blocked the nuclear translocation of P65. In mice, API alleviated corneal edema and inflammatory cell infiltration and decreased IL-6 and TNF-α levels in corneal tissue. CONCLUSION:API effectively attenuates LPS-induced corneal inflammation by inhibiting JNK/ERK and NF-κB signaling pathways and downstream effectors, thereby reducing the production of pro-inflammatory cytokines such as IL-6, IL-8, COX2, and TNF-α.

Immune-mediated graft rejection (IMGR) remains a leading cause of corneal transplant (CT) failure in high-risk cases. While transcorneal delivery of tacrolimus (FK506) is a promising strategy, the cornea’s barrier properties severely limit FK506′s bioavailability and therapeutic function. Here, we developed a transcorneal delivery system by loading FK506 into zeolitic imidazolate framework-8 (ZIF-8) to form the FK506@ZIF-8 nanoparticles (<100 nm) with 4.5 % drug-loading capacity, followed by mixing with 2-hydroxypropyltrimethyl ammonium chloride chitosan (HTCC) solution to form a homogeneous sol. In vitro studies demonstrated sustained FK506 release over 24 h of the FK506@ZIF-8 in a simulated ocular microenvironment via ZIF-8 framework hydrolysis (Weibull model) and confirmed its biosafety in human corneal epithelial cells (HCECs) at concentrations ≤ 20 μg/mL. The Draize test validated minimal ocular irritation of the FK506@ZIF-8/HTCC eye drop in rabbits. The nano-size of the FK506@ZIF-8 combined with the HTCC-mediated corneal adhesion (>240 min retention) synergistically enhanced transcorneal delivery of FK506. In a rabbit IMGR model, the FK506@ZIF-8/HTCC eye drop significantly prolonged graft survival, reduced inflammatory infiltration, and outperformed conventional FK506 eye drops, highlighting its therapeutic potential for overcoming corneal drug delivery barriers. In summary, the FK506@ZIF-8/HTCC sol hold great promise for preventing high-risk keratoplasty rejection in clinic in the future.

Objectives: To characterise the bacterial and fungal spectrum of infectious keratitis (IK) in southern China and to evaluate changes in the bacterial profiles and antimicrobial resistance (AMR) over an 8-year period (2017-2024). Methods: This retrospective study included patients with culture-positive IK treated between 2017 and 2024. Corneal scrapings were obtained for microbiological culture and pathogen identification. Antimicrobial susceptibility testing was performed for all bacterial isolates. Microbial distribution and in vitro antibiotic susceptibility were analysed. Results: A total of 2785 microbial isolates were recovered from 2741 patients. Overall, fungal isolates predominated (59.6%), exhibiting a distinct seasonal distribution, with Fusarium (40.2%) and Aspergillus (14.3%) being the most common genera. Among bacterial isolates, Gram-positive organisms were predominant (63.6%). The most frequently identified Gram-positive organisms were coagulase-negative staphylococci (CNS; 34.9%), while Pseudomonas (18.9%) was the most common Gram-negative pathogen. Over the study period, an increase in the proportions of CNS (p < 0.001) and Serratia was observed (p = 0.017), alongside a decline in Pseudomonas (p = 0.009) and Kocuria (p < 0.001). Resistance among Gram-positive isolates increased for penicillin (from 62.3% to 74.4%; p = 0.002) and levofloxacin (from 26.4% to 46.9%; p < 0.001), whereas Gram-negative resistance generally declined. Conclusions: IK in southern China is characterised by persistent fungal predominance and evolving bacterial composition. AMR patterns differ between Gram-positive and Gram-negative organisms, reflecting both shifts in pathogen distribution and species-specific resistance changes, highlighting the importance of continued regional surveillance to guide empirical therapy.

Currently, research on optic nerve injury predominantly focuses on the retina and optic nerve, but emerging evidence suggests that optic nerve injury also affects advanced visual structures like the superior colliculus (SC) and primary visual cortex (V1 region). However, the exact mechanisms have not been fully explored. This study aims to investigate the characteristics and mechanisms of pathology in the SC and V1 region after optic nerve crush (ONC) to deepen our understanding of the central mechanism of visual injury. After unilateral ONC, visual acuity in the injured eye declined, along with thinning of the retinal nerve fiber layer, and the latency and amplitude of FVEPs decreased. Furthermore, neuronal loss and degeneration were observed in the contralateral SC and V1 region, accompanied by astrocytic activation. Additionally, protein markers C3, and Serping1 for A1 astrocytes, which had neurotoxic effects and S100A10, and PTX3 for A2 astrocytes, which promoted tissue repair, were increased in the two regions. A1 astrocytes were mainly present in the early stages of observation, while A2 astrocytes were mainly increased later. Notably, NLRC4, GSDMD-N, cleaved caspase-1 expression, and IL-1β, IL-18 secretion increased in the contralateral SC and V1 region. Collectively, our findings reveal that A1 (neurotoxic) and A2 astrocytes (neuroprotective), NLRC4-mediated neuronal pyroptosis are enhanced in SC and V1 region contralateral to the ONC eye. The primary visual cortex responds to injury later than the superior colliculus after ONC, with less pronounced damage changes. Reactive astrocytes and NLRC4 inflammasome may act as promising targets for the prevention and treatment of optic nerve injury.

OBJECTIVE:Glaucoma filtration failure may result from an overabundance of human Tenon's capsule fibroblasts (HTFs) forming a filtration tract scar. Conversely, the Yes-associated protein (YAP), a transcriptional activator of the Hippo signaling pathway, is a crucial matrix stiffness regulator of matrix production and fibroblast activation. With superior biocompatibility and biodegradability, RGD peptide hydrogels imitate the structure of real tissues' extracellular matrix (ECM). The purpose of this research was to determine whether down-regulating YAP expression via RGD peptide hydrogels may prevent HTFs activation and ECM protein secretion. Transforming growth factor-β2 (TGF-β2) was used to induce the activation of HTFs in a cellular model of scarring following glaucoma filtration surgery. Utilizing SD rats, a murine model of subconjunctival injury was established. The shape of collagen fibers was observed through Masson staining, and the expression of YAP and α-smooth muscle actin (α-SMA) was identified through immunohistochemistry. RGD peptide hydrogel was discovered to have anti-scarring properties in a mouse eye injury model, as well as the ability to lessen HTFs activation, YAP expression, cytosolic nucleus accumulation, and the expression of connective tissue growth factor (CTGF) and ECM proteins. The best concentration was found to be 1.0 weight percent among them. This concentration not only makes it easier to inject a drug subconjunctivally in vivo and maintain the filtration vesicle space in the conjunctiva, but it also inhibits the activation of fibroblasts into myofibroblasts and down-regulates the expression of the Hippo-YAP signaling pathway in Tenon's capsule fibroblasts. STATEMENT OF SIGNIFICANCE:1. The homogenous reticular three-dimensional nanostructure that made up the interior structure of the 1.0 weight percent gel had good drug delivery characteristics for long-lasting controlled drug release. 2. RGD peptide hydrogel had a certain matrix hardness, which could mimic the normal connective tissue hardness under the conjunctiva. 3. RGD peptide hydrogels could prevented the development of rat conjunctival fibrosis. 4. RGD peptide hydrogel could inhibit the expression of YAP and its target gene CTGF, as well as α-SMA, ECM proteins in HTFs. 5. RGD peptide hydrogel has good biocompatibility, biodegradability, and stable mechanical properties, and can also be used as a promising carrier for the controlled release of drugs.

This study aimed to investigate whether Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) could identify Herpes simplex virus type 1 (HSV1) infection in samples in vitro and in vivo. MS spectra of supernatants and suspensions from infected human cornea epithelial (HCE) cell culture samples and infected samples of BALB/c mouse corneas were obtained by a VITEK® mass spectrometer. The discriminating peaks between infected and non-infected samples were used to establish discriminating superspectra (DSPc for cells and DSPm for corneas) by SARAMIS™ software. Another infected cells with two viral titers and infected cornea samples were used for blind testing against two DSPs. The results showed that automatic matching by the SARAMIS system revealed 28 discriminating peaks in HSV1-infected cells and 17 discriminating peaks in HSV1 keratitis, generating two discriminating superspectra (DSPs). Blind testing of virus-infected samples demonstrated a high positive identification rate for both in vitro and in vivo DSPs. The positive identification rate varied with viral titers, with cell suspensions exhibiting significantly higher rates compared to supernatants. Cluster analysis based on MS spectra revealed that there were more obvious differences between in vivo and in vitro samples compared to the differences between infected and non-infected samples. These findings suggest that MALDI-TOF MS can directly identify HSV1 in vitro or in vivo infected specimens, with higher positivity rates achieved when using cellular suspensions directly. This is an attempt on the method of virus detection, which shows potential for using MS to detect HSV1 infection or other virus infection in humans.

Herpes simplex keratitis (HSK) is a recurrent inflammatory disease of cornea primarily initiated by type I herpes simplex virus infection of corneal epithelium. However, early diagnosis of HSK remains challenging due to the lack of specific biomarkers. This study aims to identify biomarkers for HSK through tear metabolomics analysis between HSK and healthy individuals. We conducted a cross-sectional study enrolling 33 participants. Tear samples were collected from one eye of 18 HSK patients and 15 healthy volunteers using Schirmer-strips. Tear metabolomic profiling was performed using high-performance liquid chromatography tandem mass spectrometry (LC–MS/MS). Metabolites were quantified and matched against entries in the human metabolome database (HMDB) and small molecule pathway database (SMPDB) to identify metabolites and metabolic pathways, respectively. Metabolic differences between HSK and control group were determined using multivariate statistical analysis. A total of 329 metabolites were identified, of which 18 were significantly altered in HSK patients. Notably, 12 metabolites were significantly increased, and 6 were significantly decreased in HSK patients. The changed metabolites were enriched in these pathways: arginine and proline metabolism, phospholipid biosynthesis, alpha linolenic acid and linoleic acid metabolism, retinol metabolism. To assess the potential utility of tear biomarkers, a predictive model was developed combining 4 metabolites (AUC = 0.998 [95

Quercetin is a natural compound with potent antiviral effects; however, its role in the treatment of herpes simplex keratitis (HSK) remains underexplored. Here, we investigated the antiviral effects of quercetin against herpes simplex virus 1 (HSV-1). By examining different phases of viral infection in human corneal epithelial cells (HCECs), we found that 30 μmol/L quercetin inhibits HSV-1 replication primarily by disrupting viral attachment. RNA-sequencing and subsequent analyses revealed that the nuclear factor E2-related factor 2 (Nrf2) was upregulated by quercetin in a dose-dependent manner. Knocking down Nrf2 partially compromised quercetin's antiviral effect. Importantly, topical application of 100 μmol/L quercetin alleviated HSK severity in mice, reduced viral titers in tears, and inhibited VP16 expression in the cornea and trigeminal ganglia. These findings demonstrate the antiviral effect of quercetin against HSV-1 and provide a foundation for mechanistic studies to elucidate its therapeutic potential in HSK.

Purpose:The purpose of this study was to investigate the characteristics of sustained drug release systems established by polylactic acid (PLLA)/RGD/mitomycin C (MMC) electrospun nanofiber membrane in vitro, and confirm its anti-scarring effect in a rabbit model of glaucoma filtration surgery (GFS). Methods:In vitro experiments: (1) PLLA/RGD/MMC nanofiber membrane drug delivery system was prepared by electrospinning technology. (2) Characterization of nanofiber membrane. (3) Biocompatibility detection of nanofiber membrane. In vivo experiment: construct a surgical model for rabbit eye GFS, divided into four groups: the control group, the PLLA/RGD membrane group, the 0.4 mg/mL MMC group, and the PLLA/RGD/MMC membrane group. The morphology of filtering blebs and wound healing were observed on the 7th, 14th, and 28th days after the operation. At 28 days after the operation, histological and immunohistochemical staining were performed to observe the scar formation. Finally, the results were statistically analyzed. Results:The composition, structure, hydrophily, thermal behaviors, degradability, and mechanical properties of the membrane were investigated in detail. In vitro cell culture assay indicated that the PLLA/RGD/MMC2 membrane could release MMC in a sustained manner for over 25 days. In vitro cell culture assay proved the superior cytocompatibility of the membrane with Human embryonic Tenon's capsule fibroblasts (HFTFs). Histology and immunohistochemistry indicated that the membrane could efficiently inhibit scaring formation after GFS and showed significant advantage over the conventional MMC cotton pad. Conclusions:The as-prepared PLLA/RGD/MMC membrane could be a potential candidate for inhibiting scaring formation after GFS in the future. Translational Relevance:This work shed some light to the developments and applications of MMC loaded electrospinning membrane for inhibiting postoperative fibrosis in GFS.

Purpose:Superior limbic keratoconjunctivitis (SLK) and dry eye disease (DED) exhibit similar clinical manifestations, complicating diagnosis. We aimed to compare tear metabolomic profiles and secretory phospholipase A2 (sPLA2) levels in patients with SLK versus patients with DED. Methods:We established a cross-sectional study involving 56 subjects: 20 patients with SLK, 21 patients with DED, and 15 matched healthy controls (solely for measuring sPLA2). Tear fluids collected via Schirmer strips underwent liquid chromatography with tandem mass spectrometry metabolomic analysis. Metabolites were quantitatively analyzed and matched to metabolic pathways and biomarkers. Metabolic differences between patients with SLK and patients with DED were identified through multivariate statistical analysis. Western blot measured the content of sPLA2 in tears of the three groups. Results:Tear analysis annotated 274 metabolites. Twenty-three metabolites showed significant differences (P < 0.05) between the 2 groups, comprising 20 increased and 3 decreased in SLK. Top differential metabolic pathways were glycerophospholipid, alpha-linolenic acid, and linoleic acid metabolism. Receiver operating characteristic (ROC) analysis showed that five metabolites could be considered as potential biomarkers for distinguishing SLK from DED. Tears PLA2 was significantly higher in SLK than healthy controls (p < 0.05), but without difference between SLK and DED. Conclusions:The study revealed distinct metabolomic profiles in SLK versus DED tears, with notable changes in glycerophospholipid metabolism and elevated sPLA2 levels in SLK. Translational Relevance:Tear metabolomics distinguishes SLK from DED at the molecular level through dysregulated glycerophospholipid metabolism. In SLK, compared with controls, this dysregulation coincides with elevated sPLA2, an enzyme hydrolyzing glycerophospholipids, indicating lipid-driven inflammation.

AIM: To explore whether plasma proteins serve as potential therapeutic targets for primary open angle glaucoma (POAG) based on a Mendelian randomization (MR) study. METHODS: Large-scale protein quantitative trait loci (pQTLs) data from the Icelandic deCODE database and two large POAG Genome-Wide Association Study (GWAS) summary datasets were used in this study. Causal associations between plasma proteins and POAG were identified using summary-data-based MR (SMR) analysis and the heterogeneity in dependent instruments (HEIDI) test. Colocalization analysis was then conducted to assess the genetic associations between these two factors. Phenotype-wide MR analysis was performed to validate protein targets as potential drug targets and to evaluate potential side effects. Finally, protein-protein interactions (PPI) were studied, and the Drug-Gene Interaction Database (DGIDb) was used to identify associations between drugs and the identified proteins. RESULTS: Four proteins (SVEP1, TMEM190, ROBO1, and ENPP5) were identified as potential drug targets in this study. Phenome-wide MR analysis showed that SVEP1, ROBO1, and ENPP5 were not associated with adverse effects, while TMEM190 was linked to nerve root and plexus disorders, as well as subarachnoid hemorrhage. Ticagrelor was suggested as a potential new drug for the treatment of glaucoma by regulating SVEP1. CONCLUSION: Four plasma proteins—SVEP1, TMEM190, ROBO1, and ENPP5—are identified as potential therapeutic targets for POAG through an MR approach. Phenome-wide MR analysis reveals that SVEP1, ROBO1, and ENPP5 are not associated with adverse effects, while TMEM190 is linked to nerve root and plexus disorders, as well as subarachnoid hemorrhage. Ticagrelor is proposed as a potential therapeutic drug for glaucoma by regulating SVEP1. These findings highlight the potential of plasma proteins as drug targets for POAG and provide valuable insights for further research.

[Objective:] To understand the infection status of four blood-borne pathogens among ophthalmology patients in South China and the distribution of positive patients across different eye diseases. [Methods:] A retrospective analysis of data from outpatient and inpatient or surgical ophthalmic patients who underwent screening for hepatitis B surface antigen (HBsAg), hepatitis C virus antibody (anti-HCV), treponema pallidum antibody (anti-TP) and human immunodeficiency virus antibody (anti-HIV) at Zhongshan Ophthalmic Center of Sun Yat-sen University from January 2017 to December 2023. The positivity rates of these markers were recorded and analyzed. [Results:] A total of 253246 patients were included in the study, with positivity rates for HBsAg, anti-HCV, anti-TP, and anti-HIV at 9.00%, 0.45%, 1.55% and 0.13%, respectively. Among these, 686 patients tested positive for two or more markers, with the highest co-positivity observed for HBsAg and anti-HCV (57.29%), followed by anti-HIV and anti-TP (15.74%). The positivity rate of HBsAg showed a yearly decline, while the rates for anti-HCV, anti-HIV, and anti-TP remained relatively stable from 2017 to 2023. Males had significantly higher positivity rates for HBsAg, anti-HCV, anti-TP, and anti-HIV compared to females (P < 0.001). Significant differences in positivity rates for these markers were also observed across different age groups (P < 0.001). In patients with positive HBsAg, anti-HCV, and anti-TP, the proportion of cases with lens diseases were the highest, at 36.72%, 39.12%, and 46.97%, respectively. Vitreoretinal diseases followed, with proportions of 22.52%, 19.65%, and 19.69%, respectively. Among patients with positive anti-HIV, the proportion of cases with vitreoretinal diseases was the highest, at 42.99%, followed by lens diseases at 26.17%, and uveitis at 8.72%. [Conclusions:] This study reveals the infection status of four blood-borne pathogens in ophthalmology patients and the distribution of infected patients across different eye diseases, which is significant for assessing the risks of ophthalmic surgery and formulating infection control measures.

Bacterial keratitis is the serious corneal infection involving inflammation, but the proinflammatory effects of Gram-positive bacterial cell wall components on human corneal epithelial cells (HCECs) remain implicit. A20 serves as a crucial regulator in many systemic or topical inflammation. Herein, we firstly explore the effects on corneal epithelial inflammation and A20 expression of different cell wall components. Immortalized HCECs were treated with various concentrations of Staphylococcus aureus cell wall components (lipoteichoic acid [LTA], peptidoglycan, and staphylococcal protein A), with lipopolysaccharide (LPS) from Pseudomonas aeruginosa as a positive control. Only LTA and LPS significantly induced the phosphorylation of p65, p38, JNK, and ERK, along with increased IL-6, IL-8, and A20 expression. LTA (10 μg/mL) elicited the proinflammatory effect similar to LPS. Thus, we further investigated the role of A20 in the LTA-mediated inflammatory injury in HCECs. The A20 expression exhibited both time- and dose-dependent manner, and peaked at 4 h after LTA treatment. Reduced A20 expression exacerbated IL-6 and IL-8 levels induced by LTA, while increased A20 augmented their expression. Pretreatment with p65, p38, JNK, and ERK antagonists downregulated A20 expression, indicating NF-κB and MAPK pathways are required for A20 expression. A20 also suppressed NF-κB and MAPK pathways activation, confirmed by both A20-knockdown and A20-overexpressed HCECs following LTA induction. These findings suggest that LTA induces the A20 expression in HCECs due to its strong immunogenicity. A20 expression is pivotal in mitigating inflammatory response to Gram-positive bacterial infections through NF-κB and MAPK pathways, and reduced IL-6 and IL-8 levels.

Cardiac fibrosis characterized by aberrant activation of cardiac fibroblasts impairs cardiac contractile and diastolic functions, inducing the progression of the disease towards its terminal phase, resulting in the onset of heart failure. Therefore, the inhibition of cardiac fibrosis has become a promising treatment for cardiac diseases. The ovarian follicle-stimulating hormone folliculin (FLCN) plays a significant role in various biological processes, such as lysosome function, mitochondrial synthesis, angiogenesis, ciliogenesis and autophagy. Severe heart failure was observed in FLCN knockout mice. In this study, we investigated the role of FLCN in cardiac fibrosis and its potential mechanisms. The mice were subjected to transverse aortic constriction (TAC) surgery. Myocardial fibrosis developed in the mice 8 weeks after surgery. We showed that the protein and mRNA expression levels of FLCN were significantly decreased in TAC mice. Similar results were observed in primary mouse cardiac fibroblasts treated with Ang-II, an in vitro cardiac fibrosis model, suggesting that FLCN is involved in the pathological process of cardiac fibrosis. We demonstrated that overexpression of FLCN inhibited lysosome function in cardiac fibroblasts. Furthermore, overexpression of FLCN protected the heart from TAC-induced pathological cardiac fibrosis. We revealed that FLCN bound to the cPLA2 protein, increased its activity, regulated lysosomal function, and promoted membrane permeabilisation in cardiac fibroblasts during cardiac fibrosis. Knockdown of cPLA2 blocked the antifibrotic effect of FLCN in cardiac fibrosis. In addition, we found that the reduced expression of FLCN in cardiac fibrosis resulted from the modulation of YTHDF3-regulated m6A methylation of FLCN mRNA. The overexpression of YTHDF3 alleviated the production of collagens and improved cardiac structure and function in TAC mice. YTHDF3 inhibited proliferation and differentiation and regulated lysosomal function in mouse cardiac fibroblasts, whereas these effects were abolished by FLCN knockdown. We conclude that FLCN undergoes YTHDF3-regulated m6A modification and interacts with cPLA2 to improve lysosomal function in cardiac fibroblasts, highlighting its role in myocardial fibrosis therapy. These results suggest that FLCN and YTHDF3 could serve as potential therapeutic targets for cardiac fibroblast treatment.

[This corrects the article DOI: 10.3389/fnmol.2023.1083850.].

Aims To explore the possibility of implementing Choosing Wisely on ocular patients in China by investigating the prevalence of abnormalities in routine preoperative blood tests (RPBTs) and its turnaround time (TAT). Methods Data from 102 542 ocular patients between January 2016 and December 2018, at Zhongshan Ophthalmic Center, were pooled from the laboratory information system. The test results were divided into normal and abnormal, including critical values. Ocular diseases were stratified into 11 subtypes based on the primary diagnosis. The TAT of 243 350 blood tests from January 2017 to December 2018 was categorised into transportation time and intralaboratory time. Results RPBT was grouped into complete blood count (CBC), blood biochemistry (BBC), blood coagulation (BCG) and blood-borne pathogens (BBP), completed for 97.22%, 87.66%, 94.41% and 95.35% of the recruited patients (male, 52 549 (51.25%); median(IQR) age, 54 (29–67) years), respectively. Stratified by the test items, 9.19% (95% CI 9.07% to 9.31%) were abnormal results, and 0.020% (95% CI 0.019% to 0.022%) were critical; most abnormalities were on the CBC, while glucose was the most common critical item. Classified by the patients’ primary diagnosis, 76.97% (95% CI 76.71% to 77.23%) had at least one abnormal result, and 0.28% (95% CI 0.25% to 0.32%) were critical; abnormal findings were reported in 45.29% (95% CI 44.98% to 45.60%), 54.97% (95% CI 54.65% to 55.30%), 30.29% (95% CI 30.00% to 30.58%) and 11.32% (95% CI 11.12% to 11.52%) for the CBC, BBC, BCG and BBP tests, respectively. The median transportation time and intralaboratory TAT of the samples were 12 min and 78 min respectively. Conclusion Blood abnormalities are common in ocular patients. With acceptable timelines, RPBT is still indispensable in China for patient safety.

This study aimed to identify and quantify free fatty acids (FFAs), secretory phospholipase A2 group IIa (sPLA2-IIa) and cytosolic phospholipase A2 (cPLA2) in serum of superior limbic keratoconjunctivitis (SLK) patients and explored the association between FFAs, sPLA2-IIa and cPLA2 variations and SLK. Targeted metabolomic analysis of FFAs in serum was performed by gas chromatography tandem mass spectrometry (GC-MS/MS) analysis on 16 SLK patients (43.88±7.88 years; female: 62.50%) and 25 healthy controls (43.12±7.88 years; female: 64.00%). Qualitative and absolute quantitative results of FFAs were obtained and classified according to gender and thyroid tests. Differential lipid metabolites, metabolomic pathways and biomarkers were further evaluated. The serum sPLA2-IIa and cPLA2 were determined by enzyme linked immunosorbent assay (ELISA).Among 40 FFAs identified, 6 FFAs showed significant changes (P<0.05) in SLK patients, including 4 decreased and 2 increased. They were mainly related to unsaturated fatty acid biosynthesis, α-linolenic acid and linoleic acid metabolism, and fatty acid biosynthesis. When dividing the data by gender or abnormal thyroid tests, some comparable FFAs alterations displayed in SLK patients. The ROC analysis revealed that the AUC values of linoleic acid, γ-linolenic acid, cis-8,11,14-eicosatrienoic acid, stearic acid, and palmitic acid, were all greater than 0.8. The serum concentrations of sPLA2-IIa and cPLA2 in patients with SLK were significantly higher than that in healthy controls. Lipidomics disturbance might be the potential mechanism of SLK. Serum FFA biomarkers associated with SLK have potential for the diagnosis and treatment of the disease.