Activation of the SST-SSTR5 signaling pathway enhances corneal wound healing in diabetic mice.

Corneal wound healing in diabetic patients is usually delayed and accompanied by excessive inflammation. However, the underlying cellular and molecular mechanisms remain poorly understood. Here, we found that somatostatin (SST), an immunosuppressive peptide produced by corneal nerve fibers, was significantly reduced in streptozotocin-induced diabetic mice. In addition, we discovered that topical administration of exogenous SST significantly improved re-epithelialization and nerve regeneration following diabetic corneal epithelial abrasion. Further analysis showed that topical SST significantly reduced the expression of injury inflammation-related genes, inhibited neutrophil infiltration, and shifted macrophage polarization from pro-inflammatory M1 to anti-inflammatory M2 in diabetic corneas' healing. Moreover, the application of L-817,818, an agonist of the SST receptor type 5 subtype, significantly reduced the inflammatory response following epithelial injury and markedly improved the process of re-epithelialization and nerve regeneration in mice. Taken together, these data suggest that activation of the SST-SST receptor type 5 pathway significantly ameliorates diabetes-induced abnormalities in corneal wound repair in mice. Targeting this pathway may provide a novel strategy to restore impaired corneal wound closure and nerve regeneration in diabetic patients.

YAP-mediated GPER signaling impedes proliferation and survival of prostate epithelium in benign prostatic hyperplasia.

Benign prostatic hyperplasia (BPH) occurs when there is an imbalance between the proliferation and death of prostate cells, which is regulated tightly by estrogen signaling. However, the role of G protein-coupled estrogen receptor (GPER) in prostate cell survival remains ambiguous. In this study, we observed that prostates with epithelial hyperplasia showed increased yes-associated protein 1 (YAP) expression and decreased levels of estrogen and GPER. Blocking YAP through genetic or drug interventions led to reduced proliferation and increased apoptosis in the prostate epithelial cells. Interestingly, GPER agonists produced similar effects. GPER activation enhanced the phosphorylation and degradation of YAP, which was crucial for suppressing cell proliferation and survival. The Gαs/cAMP/PKA/LATS pathway, downstream of GPER, transmitted signals that facilitated YAP inhibition. This study investigated the interaction between GPER and YAP in the prostate epithelial cells and its contribution to BPH development. It lays the groundwork for future research on developing BPH treatments.

Exploring Immune Cell Diversity in the Lacrimal Glands of Healthy Mice: A Single-Cell RNA-Sequencing Atlas.

The lacrimal gland is responsible for maintaining the health of the ocular surface through the production of tears. However, our understanding of the immune system within the lacrimal gland is currently limited. Therefore, in this study, we utilized single-cell RNA sequencing and bioinformatic analysis to identify and analyze immune cells and molecules present in the lacrimal glands of normal mice. A total of 34,891 cells were obtained from the lacrimal glands of mice and classified into 18 distinct cell clusters using Seurat clustering. Within these cell populations, 26 different immune cell subpopulations were identified, including T cells, innate lymphocytes, macrophages, mast cells, dendritic cells, and B cells. Network analysis revealed complex cell-cell interactions between these immune cells, with particularly significant interactions observed among T cells, macrophages, plasma cells, and dendritic cells. Interestingly, T cells were found to be the main source of ligands for the Thy1 signaling pathway, while M2 macrophages were identified as the primary target of this pathway. Moreover, some of these immune cells were validated using immunohistological techniques. Collectively, these findings highlight the abundance and interactions of immune cells and provide valuable insights into the complexity of the lacrimal gland immune system and its relevance to associated diseases.

Regulation of Interferon-ß-Modified Human Umbilical Cord Mesenchymal Stem Cell-Derived Exosomes in Proliferation and Apoptosis of Prostate Cancer Cells.

Prostate cancer (PCa) poses a serious burden to men. Interferon-ß (IFN-ß) is implicated in cancer cell growth. This study hence explored the regulation of IFN-ß-modified human umbilical cord mesenchymal stem cell-derived exosomes (hUCMSC-Exos) in PCa cells. In vitro-cultured hUCMSCs were transfected with pcDNA3.1-IFN-ß plasmid or IFN-ß siRNA. hUCMSC-Exos were extracted by ultracentrifugation and identified. PCa cells (PC3 and LNCap) were treated with Exos. Cellular internalization of Exos by cells was detected by uptake assay. Cell proliferation, cycle, and apoptosis were evaluated by CCK-8, EdU staining, and flow cytometry. Levels of cell cycle-related proteins (cyclin D/cyclin E) were determined by Western blot. The effect of IFN-ß-modified hUCMSC-Exos in vivo was analyzed. IFN-ß-modified hUCMSC-Exos (Exooe-IFN-ß or Exosi-IFN-ß) were successfully isolated. IFN-ß was encapsulated in Exos, and PCa cells could uptake Exos. After treating with Exooe-IFN-ß, PCa cell proliferation was impeded, the percentage of cells in the G0/G1 phase, cyclin D/cyclin E levels, and cell apoptotic rate were elevated, while cells treated with Exooe-IFN-ß exhibited contrary trends. IFN-ß-modified hUCMSC-Exos reduced PCa tumor size and weight in vivo. Conjointly, IFN-ß-modified hUCMSC-Exos suppress PCa cell proliferation and facilitate apoptosis.

The association between fasting blood glucose and prognosis in gastrointestinal stromal tumor patients after curable resection.

Blood glucose has been demonstrated to serve as prognostic indicators in various malignancies. This study aimed to explore the relationship between fasting blood glucose (FBG) levels and the prognosis in patients with gastrointestinal stromal tumor (GIST) undergoing complete resection. Data were retrospectively collected from 256 patients with primary GIST underwent complete surgical resection or endoscopic excision. Patients were stratified into euglycemic group and hyperglycemic group. Patients' characteristics between groups were compared. Cox regression model was conducted to identify independent prognostic factors of disease-free survival (DFS). Both univariate analysis and multivariate analyses revealed that FBG ≥ 100 mg/dl was associated with poor outcomes. Patients with FBG ≥ 100 mg/dl tended to have more adverse features, more likely to suffer recurrence and a worse 5-year DFS than patients with FBG < 100 mg/dl. Moreover, FBG levels helped distinguishing between patients with different survival outcomes in different risk categories defined by modified NIH systems. Our data provided the evidence that FBG is a useful prediction marker prognosis in patients with GIST undergoing curative surgery.

GLIDR promotes the aggressiveness progression of prostate cancer cells by sponging miR-128-3p.

Glioblastoma downregulated RNA (GLIDR) is a newly discovered long non-coding RNA (lncRNA) that its increased expression indicates a poor prognosis of prostate cancer (PCa). However, the effect of GLIDR on PCa cells is not clear. Our study investigated the role and molecular mechanism of GLIDR in PCa cells. The results showed that GLIDR expression levels were higher in PCa samples and cells than in control. GLIDR could regulate the invasive potential, epithelial-to-mesenchymal transition (EMT) and proliferation in PC-3 and LnCaP cells. Besides, GLIDR could weaken the inhibitory effects of miR-128-3p on invasion, EMT and proliferation in PCa cells. Western blotting proved that miR-128-3p affected the expression of EMT markers, such as E-cadherin, Snail and N-cadherin, and GLIDR could reversed the effects of miR-128-3p on the expression levels of EMT markers in PCa cells. In addition, knockdown of miR-128-3p stimulated the invasion, EMT, and proliferation in PCa cells, whereas these effects were reversed when GLIDR expression was knocked down. GLIDR knockdown inhibited the invasion, EMT, and proliferation in PCa cells, and GLIDR was shown to sponge miR-128-3p. Together, these results highlight GLIDR as a potential therapeutic target for the PCa treatment.

A validated tool of combining risk factors with fecal immunochemical test for selecting colonoscopy in asymptomatic Chinese population.

Fecal immunochemical test (FIT) is often used for preselection for colonoscopy, but FIT has nonoptimal sensitivity. Selection of study participant for colonoscopy based on the result of combining FIT with risk factors could improve the sensitivity of a screening program. We aimed to develop a risk prediction system of colorectal neoplasia among asymptomatic Chinese subjects. A total of 6265 asymptomatic participants with age between 50 and 70 years were invited to undergo colonoscopy screening. They were also asked to take FIT and complete a questionnaire for collecting information on risk factors. Independent risk factors were identified by binary logistic regression for colorectal neoplasia. A risk score model was developed by using the odds ratios of significant risk factors. The scoring system was divided into two groups of risk: negative risk and positive risk. The performance of the risk score model in terms of predicting colorectal neoplasia was evaluated. Of the 1786 colonoscopy screening participants, 1546 completed FIT and questionnaires. A total of 462 cases of neoplasia were detected. Based on the scoring stratification, 966 (62.5%) participants were in negative risk tier and 580 (37.5%) were in positive risk tier. The incidence of colorectal neoplasia in negative risk and positive risk groups was 18.4 and 49.0%, respectively. Risk stratification model has better ability to discriminate those with or without colorectal neoplasia than FIT-only model. Classification improved significantly with risk stratification-based screening (net reclassification improvement = 0.064, P = 0.032). Risk stratification system increases the predictive value of FIT-based screening and is useful for preselection for colonoscopy in colorectal cancer screening program.

Recent advances in graves ophthalmopathy medical therapy: a comprehensive literature review.

Graves ophthalmopathy (GO), which occurs in autoimmune thyroid disease, can reduce patients' quality of life due to its impact on visual function, physical appearance, and emotional health. Corticosteroids have been the first-line treatment for GO. More recently, the pathogenesis of GO has made significant progress. Various targeting biological agents and immunosuppressive agents make GO management more promising. Fully understanding GO pathogenesis and precise clinical management are beneficial for the prognosis of patients. Therefore, we conducted a comprehensive review of the medical management of GO and summarized research developments to highlight future research issues.

Reproducibility of Three-Dimensional Volumetric Measurement of Periocular Tumor Models.

INTRODUCTION: In this study, we measured the volume of customized tumor models in the periocular area using three-dimensional (3D) stereophotogrammetry and evaluated the reproducibility of these measurements. METHODS: Five tumor models of different colors and sizes were placed in different periocular positions, and 3D facial images were obtained from 68 healthy adult volunteers. Subsequently, the volumes of the tumor models were measured, and the intra- and interrater reproducibility was assessed. RESULTS: The gray 6 mm model revealed the highest reliable measurements in both Caucasians (intra- and interrater intraclass correlation coefficients of 0.981 and 0.899, mean absolute difference of 1.446 and 3.327 mm3, relative error measurement of 3.497% and 8.120%, technical error of measurement of 1.450 and 3.105 mm3, and relative technical error of measurement of 3.506% and 7.580%) and Asians (0.968 and 0.844, 1.974 and 4.067 mm3, 4.772% and 9.526%, 2.100 and 4.302 mm3, and 5.076% and 10.076%, respectively). The highest reliability of measurements in the lateral upper eyelid (0.88 and 0.95, 4.042 and 3.626 mm3, 9.730% and 9.020%, 5.714 and 3.358 mm3, and 9.730% and 8.350%, respectively) and medial upper eyelid (0.81 and 0.89, 4.313 and 4.226 mm3, 9.730% and 9.020%, 6.098 and 4.069 mm3, and 9.730% and 8.350%, respectively) with eyes closed was evident in Caucasians, while the same trend (0.841 and 0.815, 2.828 and 3.757 mm3, 9.860% and 9.840%, 4.052 and 4.308 mm3, and 9.860% and 9.740%, respectively) was observed in Asians in the medial canthus with eyes closed. CONCLUSIONS: This study confirms, for the first time, the high reliability of periocular tumor volume measurements using 3D stereophotogrammetry, suggesting its feasibility for eyelid tumor measurement. Further trials are required to investigate its clinical use for documentation and follow-up of different eyelid tumors.

Age-related changes of the periocular morphology: a two- and three-dimensional anthropometry study in Caucasians.

PURPOSE: To determine age-and sex-related changes in periocular morphology in Caucasians using a standardized protocol. METHODS: Healthy Caucasian volunteers aged 18-35 and 60-90 years old were recruited from the Department of Ophthalmology, Faculty of Medicine and University Hospital, Cologne, between October 2018 and May 2020. Volunteers with facial asymmetry, facial deformities, history of facial trauma, facial surgery, botox injection, eyelid ptosis, strabismus, or nystagmus, were excluded. Standardized three-dimensional facial photos of 68 young volunteers and 73 old volunteers were taken in this clinical practice. Position changes of endocanthion, pupil center, and exocanthion were analyzed in different age and gender groups, including palpebral fissure width (PFW): distance between endocanthions (En-En), pupil centers (Pu-Pu), exocanthions (Ex-Ex), endocanthion and nasion (En-Na), pupil center and nasion (Pu-Na), exocanthion and nasion (Ex-Na), endocanthion and pupil center (Pu-En), exocanthion and pupil center (Pu-Ex), and palpebral fissure inclination (PFI); angle of endocanthions to nasion (En-Na-En), pupils to nasion (Pu-Na-Pu), exocanthions to nasion (Ex-Na-Ex); endocanthion inclination (EnI), and exocanthion inclination (ExI). RESULTS: PFW, En-En, Ex-Na, Pu-Ex, PFI, ExI, and Ex-Na-Ex were significantly different between the young and old groups (p ≤ 0.004). There were sex-related differences in PFW, Ex-Ex, En-Na, Pu-Na, Ex-Na, Pu-En, PFI, and EnI between both groups (p ≤ 0.041). CONCLUSION: The position change of the pupil is minimal relative to age; it is preferred to establish the reference plane to describe periocular changes. The endocanthion tends to move temporally and inferiorly, while the exocanthion tends to shift nasally and inferiorly with age.

A modified 3D stereophotogrammetry-based distraction test for assessing lower eyelid tension.

AIM: To investigate the reliability of a modified three-dimensional distraction test (3D-DT) and three-dimensional pinch test (3D-PT) for assessing lower eyelid tension (LET). METHODS: A cross-sectional study was conducted among 97 volunteers including 97 eyelids with no history of trauma, tumor, or reconstructive surgeries. Six three-dimensional photographs were acquired for each participant, including two photographs obtained in a neutral position (NP), two using a modified 3D-DT with a 15.9-grammes stainless steel eyelid hook performed, and two using 3D-PT. RESULTS: The mean absolute differences between NP, 3D-DT, and 3D-PT measurements varied between 0.07 and 7.42, 0.10 and 13.10, and 0.07 and 15.97, respectively; technical error of measurement varied between 0.05 and 7.81, 0.09 and 10.19, and 0.07 and 12.47, respectively; and relative error measurements varied between 0.10% and 11.50%, 0.16% and 30.51%, and 0.11% and 38.75%, respectively. For intra-rater reliability, the intraclass correlation coefficients (ICCs) were more than 0.80 in seven out of eight measurements obtained in the NP and 3D-DT, whereas those obtained in the 3D-PT were as low as less than 0.30 by rater 1; the ICCs of all the measurements obtained in all the positions (NP, 3D-DT, and 3D-PT) were more than 0.80 by rater 2. For inter-rater reliability six out of eight NP and 3D-DT measurements had an ICC greater than 0.80, whereas those of 3D-PT measurements were less than 0.30. For intra-method reliability, the ICCs of all the NP measurements were more than 0.87, whereas those of the six 3D-DT measurements and four 3D-PT measurements were more than 0.80. CONCLUSION: Our study results prove that the modified 3D-DT, which involves the use of an eyelid hook, can be a highly reliable method for evaluating LET. Furthermore, this novel and simple method may be utilized as the basis for further investigation and routine pre- and postoperative clinical evaluation.

Vector analysis of astigmatic correction after single-step transepithelial photorefractive keratectomy and femtosecond-assisted laser in-situ keratomileusis for low to moderate myopic astigmatism.

Purpose: This study aimed to evaluate the outcomes of astigmatic correction by single-step transepithelial photorefractive keratectomy (TransPRK) and femtosecond-assisted laser in-situ keratomileusis (Femto-LASIK) surgeries. Methods: A total of 218 subjects received TransPRK or Femto-LASIK surgery for the treatment of myopia and astigmatism (-2.25 to -0.25 D). Refraction errors and uncorrected (UDVA) and corrected distance visual acuity (CDVA) were examined before and at 3 months after surgery. Astigmatism changes were assessed by vector analysis. Results: Preoperative parameters of the TransPRK group were similar to the Femto-LASIK group. UDVA and CDVA at 3 months were similar between both groups. Manifest refraction (MR) spherical equivalent in the TransPRK group (0 ± 0.20 D) was slightly lower compared with the Femto-LASIK group at 3 months (0.11 ± 0.25 D, P = 0.001). MR cylinder was -0.06 ± 0.19 D in the TransPRK group and -0.02 ± 0.15 D in the Femto-LASIK group at 3 months (P = 0.135). The index of success (IS) was 0.15 ± 0.36 in the TransPRK group and 0.06 ± 0.17 in the Femto-LASIK group (P = 0.125). The correction index (CI) was 1.03 ± 0.19 in the TransPRK group and 1.01 ± 0.11 in the Femto-LASIK group (P = 0.815). Conclusion: For low to moderate myopic astigmatism, TransPRK provided a comparable astigmatic treatment effect as Femto-LASIK. Myopic astigmatism was both slightly overcorrected after TransPRK and Femto-LASIK surgeries.

Validation of the Portable Next-Generation VECTRA H2 3D Imaging System for Periocular Anthropometry.

Purpose: Portable three-dimensional imaging systems are becoming increasingly common for facial measurement analysis. However, the reliability of portable devices may be affected by the necessity to take three pictures at three time points. The purpose of this study was to evaluate the effectiveness of portable devices for assessing the periocular region. Methods: In 60 Caucasian volunteers (120 eyes), four facial scans (twice for each instrument) using the portable VECTRA H2 and static VECTRA M3 devices were performed; patients' heads were kept straight, looking ahead, with a neutral facial expression. One assessor set 52 periocular landmarks in the periocular area of each image and subsequently assessed intra- and inter-device reliability by comparing two within-device measurements and one between-device measurement, respectively. Results: The mean absolute difference (MAD) (0.13 and 0.12 units), relative error of measurement (REM) (0.61 and 0.68%), technical error of measurement (TEM)(1.02 and 0.80 units), relative TEM (rTEM) (5.51 and 4.43%), and intraclass correlation coefficient (ICC) (0.89, 0.89) showed good intra-device reliability for M3 and H2; MAD (0.63, 0.62 units), REM (2.83, 2.69%), TEM (1.31, 1.10 units), rTEM (7.62, 5.57%), and ICC (0.79, 0.83) indicated that inter-device reliability deteriorated compared to intra-device reliability and that the inter-device reliability of the first scan (moderate) was lower than that of the average of the two scans (good). Conclusions: The portable VECTRA H2 device proved reliable in assessing most periocular linear distances, curve distances, and angles; some improvement in inter-device reliability can be achieved by using the average of two scans.

Split stoma with delayed anastomosis may be preferred for 2-stage surgical resection in high-risk patients with Crohn's disease.

BACKGROUND: Fecal diversion after bowel resection is a safe and effective procedure in high-risk patients with Crohn's disease, but the better approach between primary anastomosis with protective stoma and split stoma with delayed anastomosis has not yet been investigated. This study aimed to compare the outcomes of these approaches in high-risk patients with Crohn's disease. METHODS: A retrospective investigation on consecutive high-risk patients with Crohn's disease was conducted at a tertiary referral hospital from August 2009 to March 2019. The primary outcomes were the overall early postoperative complications and overall anastomosis-related adverse events in an intention-to-treat approach. RESULTS: A total of 118 consecutive patients who underwent 121 surgeries (35 procedures with a protective stoma and 86 procedures with a split stoma) were enrolled. After a median follow-up period of 659 days and 728 days, respectively, 25 patients underwent a stoma-reversal procedure in the protective-stoma group, and 54 patients underwent delayed anastomosis in the split stoma group. Overall, early 30-day surgical morbidity and anastomosis-related adverse events were observed in more patients in the protective-stoma group than in the split-stoma group (51.4% [18/35] vs 30.2% [26/86]; P = .028 and 37.1% [13/35] vs 2.3% [2/86]; P < .001, respectively; intention-to-treat analysis). Similar results were found in the per-protocol analysis (44.0% [11/25] vs 20.4% [11/54]; P = .029 and 36.0% [12/25] vs 3.7% [2/54]; P < .001, respectively.) CONCLUSION: Split stoma with delayed anastomosis is associated with a reduction in anastomotic adverse events and overall early surgical complications and thus may be a better surgical option for high-risk patients with Crohn's disease.

Mapping research trends of uveal melanoma: a bibliometric analysis.

BACKGROUND: To explore the research trends for uveal melanoma with bibliometric methods using Web of Science Core Collection (WoSCC) and PubMed (PM). METHODS: To find UM-related studies, "uveal melanoma" was used as search term in the WoSCC and PM for the period time from 2000 to 2020. Bibliographic coupling analysis was used to investigate the journals with the highest number of UM-related publications. VOSviewer (VV) was used for mapping the knowledge domain and visualizing the co-occurrence of terms, authors, organizations, countries, co-citation literature, and keywords. The knowledge map based on WoSCC and PM was compared. RESULTS: In the WoSCC 3,748 articles were found, while in PM the search resulted in 3,403 articles. The number of original articles has steadily grown in general in the past two decades. The top ten authors were contributing to 23% (n = 856) of all publications, while the top 10 institutions published 41% (n = 1524) of all articles. The top 3 journals with the highest number of publications for UM-related research included Investigative ophthalmology & visual science, Ophthalmology, and British Journal of Ophthalmology. Co-occurrence analysis based on author keywords showed 6 clusters. The most frequent keywords included are metastasis, prognosis, and brachytherapy. The latest research hotspots focused on BAP1, immunotherapy and GNAQ. CONCLUSIONS: Genetics and immunology are the latest research frontiers in uveal melanoma. There is a clear need for interdisciplinary, molecular and clinical research approaches to improve the fatal prognosis of uveal melanoma patients.

Transcriptional regulation and ubiquitination-dependent regulation of HnRNPK oncogenic function in prostate tumorigenesis.

BACKGROUND: Heterogeneous nuclear ribonucleoprotein K (HnRNPK) is a nucleic acid-binding protein that regulates diverse biological events. Pathologically, HnRNPK proteins are frequently overexpressed and clinically correlated with poor prognosis in various types of human cancers and are therefore pursued as attractive therapeutic targets for select patients. However, both the transcriptional regulation and degradation of HnRNPK in prostate cancer remain poorly understood. METHODS: qRT-PCR was used to detect the expression of HnRNPK mRNA and miRNA; Immunoblots and immunohistochemical assays were used to determine the levels of HnRNPK and other proteins. Flow cytometry was used to investigate cell cycle stage. MTS and clonogenic assays were used to investigate cell proliferation. Immunoprecipitation was used to analyse the interaction between SPOP and HnRNPK. A prostate carcinoma xenograft mouse model was used to detect the in vivo effects of HnRNPK and miRNA. RESULTS: In the present study, we noted that HnRNPK emerged as an important player in the carcinogenesis process of prostate cancer. miR-206 and miR-613 suppressed HnRNPK expression by targeting its 3'-UTR in PrCa cell lines in which HnRNPK is overexpressed. To explore the potential biological function, proliferation and colony formation of PrCa cells in vitro and tumor growth in vivo were also dramatically suppressed upon reintroduction of miR-206/miR-613. We have further provided evidence that Cullin 3 SPOP is a novel upstream E3 ubiquitin ligase complex that governs HnRNPK protein stability and oncogenic functions by promoting the degradation of HnRNPK in polyubiquitination-dependent proteolysis in the prostate cancer setting. Moreover, prostate cancer-associated SPOP mutants fail to interact with and promote the destruction of HnRNPK proteins. CONCLUSION: Our findings reveal new posttranscriptional and posttranslational modification mechanisms of HnRNPK regulation via miR-206/miR-613 and SPOP, respectively. More importantly, given the critical oncogenic role of HnRNPK and the high frequency of SPOP mutations in prostate cancer, our results provide a molecular rationale for the clinical investigation of novel strategies to combat prostate cancer based on SPOP genetic status.

A novel standardized distraction test to evaluate lower eyelid tension using three-dimensional stereophotogrammetry.

BACKGROUND: Standardized pre-operative assessment of the lower eyelid tension is essential to determine the optimal surgical technique. However, quantitative analysis using the conventional distraction test is inaccurate and user-dependent. Our purpose was to introduce a novel, standardized three-dimensional distraction test for measuring lower eyelid tension and to determine its standard values in a Caucasian population. METHODS: In 94 participants (50 men and 44 women; age 21-85 years), a 15.9-g weighted eyelid hook was used to pull down the lower eyelid. Two three-dimensional images were acquired with a VECTRA M3 stereophotogrammetry device-one in the neutral position without a hook and the other in the distracted position with the eyelid hook. The images of all participants in both positions were measured twice by a single observer. RESULTS: There was no clinical (>1 mm) or statistically significant difference between the two repeated measurements of all the inter-landmark linear distances in both positions (P≥0.05, respectively). The mean distracted displacement between the neutral and distracted position for margin reflex distance was 5.50±1.53 mm, without any age-specific difference (P=0.08); however, a significant gender-specific difference was observed as men had significantly greater displacement than women (P<0.001). CONCLUSIONS: Our proposed standardized three-dimensional distraction test for assessing lower eyelid tension using an eyelid hook and a simple landmark-based system seems to provide high reliability. This novel and simple method might be helpful for the preoperative planning of eyelid surgeries.