Protocol to examine immune subpopulations in murine conjunctiva and lacrimal gland using flow cytometry.

Here, we present a protocol for the examination of immune cells in the murine conjunctiva and lacrimal gland using flow cytometry. We describe steps for dissection, preparation of high-quality single-cell suspensions, utilization of comprehensive staining panels, and optimization of flow cytometry voltage. We then detail procedures for compensation adjustments and the implementation of effective gating strategies. For complete details on the use and execution of this protocol, please refer to Ma et al.1.

Seeking information about waste-to-energy incineration projects: The role of objective knowledge and benefit perceptions in an extended PRISM.

Although waste-to-energy (WtE) incineration projects have boosted the economic effectiveness of the waste management system, locals frequently view them with suspicion, opposition, or even outright rejection because of potential environmental and health risks. In this study, by incorporating two additional variables, namely, objective knowledge and benefit perceptions, the planned risk information seeking model (PRISM) was extended in the context of the WtE incineration project. A total of 1726 respondents were interviewed in person. Results from the structural equation modeling indicated that the extended PRISM achieved excellent fits, suggesting the generalizability of the model in these contexts. Objective knowledge and subjective knowledge have a positive and direct impact on perceived knowledge insufficiency. The impact of benefit and risk perceptions on worry is the opposite, with risk perceptions having a greater direct effect on worry. However, compared with risk perceptions, the direct effect that benefit perceptions have on perceived knowledge insufficiency is stronger. Additionally, perceived knowledge insufficiency partially or completely mediates the relationships between all antecedents, except perceived seeking control, and information-seeking intention. This study suggests that risk managers carefully consider how to establish powerful communication tactics to shape these antecedents when introducing a "not in my backyard" project to the local community.

Analysis of Clinical Characteristics and Neuropeptides in Patients with Dry Eye with and without Chronic Ocular Pain after FS-LASIK.

INTRODUCTION: Chronic ocular pain, particularly prevalent in patients with dry eye disease and post-femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK) surgery, presents with unclear clinical characteristics and an undefined pathogenesis. In this study, we aimed to compare clinical characteristics and tear neuropeptide concentrations in patients with dry eye disease (DED) with and without chronic ocular pain following FS-LASIK, and investigate correlations between ocular pain, clinical characteristics, and tear neuropeptide levels. METHODS: Thirty-eight post-FS-LASIK patients with DED were assigned to two groups: those with chronic ocular pain and those without chronic ocular pain. Dry eye, ocular pain, and mental health-related parameters were evaluated using specific questionnaires and tests. The morphology of corneal nerves and dendritic cells (DCs) was evaluated by in vivo confocal microscopy. Function of corneal innervation was evaluated by corneal sensitivity. Concentrations of tear cytokines (interleukin [IL]-6, IL-23, IL-17A, and interferon-γ) and neuropeptides (α-melanocyte-stimulating hormone, neurotensin, ß-endorphin, oxytocin, and substance P [SP]) were measured using the Luminex assay. RESULTS: Most patients with chronic ocular pain experienced mild to moderate pain; the most common types included stimulated pain (provoked by wind and light), burning pain, and pressure sensation. More severe dry eye (P < 0.001), anxiety symptoms (P = 0.026), lower Schirmer I test values (P = 0.035), lower corneal nerve density (P = 0.043), and more activated DCs (P = 0.041) were observed in patients with ocular pain. Tear concentrations of SP and oxytocin were significantly higher in patients with ocular pain (P = 0.001, P = 0.021, respectively). Furthermore, significant correlations were observed among ocular pain severity, SP, and anxiety levels. CONCLUSIONS: Patients with DED after FS-LASIK who have chronic ocular pain show more severe ocular and psychological discomfort and higher tear levels of neuropeptides. Furthermore, ocular pain severity is correlated with tear SP levels. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT05600985.

Efficacy of topical 0.05% cyclosporine A and 0.1% sodium hyaluronate in post-refractive surgery chronic dry eye patients with ocular pain.

BACKGROUND: The management of post-refractive surgery dry eye disease (DED) can be challenging in clinical practice, and patients usually show an incomplete response to traditional artificial tears, especially when it is complicated with ocular pain. Therefore, we aim to investigate the efficacy of combined topical 0.05% cyclosporine A and 0.1% sodium hyaluronate treatment in post-refractive surgery DED patients with ocular pain unresponsive to traditional artificial tears. METHODS: We enrolled 30 patients with post-refractive surgery DED with ocular pain who were unresponsive to traditional artificial tears. Topical 0.05% cyclosporine A and 0.1% sodium hyaluronate were used for 3 months. They were evaluated at baseline and 1 and 3 months for dry eye and ocular pain symptoms and objective parameters, including Numerical Rating Scale (NRS), Neuropathic Pain Symptom Inventory modified for the Eye (NPSI-Eye), tear break-up time (TBUT), Schirmer I test (SIt), corneal fluorescein staining (CFS), corneal sensitivity, and corneal nerve morphology. In addition, tear levels of inflammatory cytokines and neuropeptides were measured using the Luminex assay. RESULTS: After 3 months of treatment, patients showed a statistically significant improvement in the ocular surface disease index (OSDI), TBUT, SIt, CFS, and corneal sensitivity (all P < 0.01) using linear mixed models. As for ocular pain parameters, the NRS and NPSI-Eye scores were significantly reduced (both P < 0.05) and positively correlated with the OSDI and CFS scores. Additionally, tear IL-1ß, IL-6, and TNF-α levels were improved better than pre-treatment (P = 0.01, 0.03, 0.02, respectively). CONCLUSION: In patients with post-refractive surgery DED with ocular pain, combined topical 0.05% cyclosporine A and 0.1% sodium hyaluronate treatment improved tear film stability, dry eye discomfort, and ocular pain, effectively controlling ocular inflammation. TRIAL REGISTRATION: Registration number: NCT06043908.

Evaluation of left ventricular stiffness with echocardiography.

Half of patients with heart failure are presented with preserved ejection fraction (HFpEF). The pathophysiology of these patients is complex, but increased left ventricular (LV) stiffness has been proven to play a key role. However, the application of this parameter is limited due to the requirement for invasive catheterization for its measurement. With advances in ultrasound technology, significant progress has been made in the noninvasive assessment of LV chamber or myocardial stiffness using echocardiography. Therefore, this review aims to summarize the pathophysiological mechanisms, correlations with invasive LV stiffness constants, applications in different populations, as well as the limitations of echocardiography-derived indices for the assessment of both LV chamber and myocardial stiffness. Indices of LV chamber stiffness, such as the ratio of E/e' divided by left ventricular end-diastolic volume (E/e'/LVEDV), the ratio of E/SRe (early diastolic strain rates)/LVEDV, and diastolic pressure-volume quotient (DPVQ), are derived from the relationship between echocardiographic parameters of LV filling pressure (LVFP) and LV size. However, these methods are surrogate and lumped measurements, relying on E/e' or E/SRe for evaluating LVFP. The limitations of E/e' or E/SRe in the assessment of LVFP may contribute to the moderate correlation between E/e'/LVEDV or E/SRe/LVEDV and LV stiffness constants. Even the most validated measurement (DPVQ) is considered unreliable in individual patients. In comparison to E/e'/LVEDV and E/SRe/LVEDV, indices like time-velocity integral (TVI) measurements of pulmonary venous and transmitral flows may demonstrate better performance in assessing LV chamber stiffness, as evidenced by their higher correlation with LV stiffness constants. However, only one study has been conducted on the exploration and application of TVI in the literature, and the accuracy of assessing LV chamber stiffness remains to be confirmed. Regarding echocardiographic indices for LV myocardial stiffness evaluation, parameters such as epicardial movement index (EMI)/ diastolic wall strain (DWS), intrinsic velocity propagation of myocardial stretch (iVP), and shear wave imaging (SWI) have been proposed. While the alteration of DWS and its predictive value for adverse outcomes in various populations have been widely validated, it has been found that DWS may be better considered as an overall marker of cardiac function performance rather than pure myocardial stiffness. Although the effectiveness of iVP and SWI in assessing left ventricular myocardial stiffness has been demonstrated in animal models and clinical studies, both indices have their limitations. Overall, it seems that currently no echocardiography-derived indices can reliably and accurately assess LV stiffness, despite the development of several parameters. Therefore, a comprehensive evaluation of LV stiffness using all available parameters may be more accurate and enable earlier detection of alterations in LV stiffness.

Transmembrane Protein CMTM6 Alleviates Ocular Inflammatory Response and Improves Corneal Epithelial Barrier Function in Experimental Dry Eye.

Purpose: To investigate the impact of transmembrane protein CMTM6 on the pathogenesis of dry eye disease (DED) and elucidate its potential mechanisms. Methods: CMTM6 expression was confirmed by database analysis, real-time polymerase chain reaction (RT-PCR), western blot, and immunohistochemistry. Tear secretion was measured using the phenol red thread test. Immune cell infiltration was assessed through flow cytometry. Barrier function was evaluated by fluorescein sodium staining, immunofluorescence staining of zonula occludens 1 (ZO-1), and electric cell-substrate impedance sensing (ECIS) assessment. For silencing CMTM6 expression, siRNA and shRNA were employed, along with lentiviral vector-mediated overexpression of CMTM6. Proinflammatory cytokine levels were analyzed by RT-PCR and cytometric bead array (CBA) analysis. Results: CMTM6 showed high expression in healthy human and mouse corneal and conjunctival epithelium but was notably reduced in DED. Notably, this downregulation was correlated with disease severity. Cmtm6-/- dry eye (DE) mice displayed reduced tear secretion, severe corneal epithelial defects, decreased conjunctival goblet cell density, and upregulated inflammatory response. Additionally, Cmtm6-/- DE mice and CMTM6 knockdown human corneal epithelial cell-transformed (HCE-T) cells showed more severe barrier disruption and reduced expression of ZO-1. Knockdown of CMTM6 in HCE-T cells increased inflammatory responses induced by hyperosmotic stress, which was significantly mitigated by CMTM6 overexpression. Moreover, the level of phospho-p65 in hyperosmolarity-stimulated HCE-T cells increased after silencing CMTM6. Nuclear factor kappa B (NF-κB) p65 inhibition (JSH-23) reversed the excessive inflammatory responses caused by hyperosmolarity in CMTM6 knockdown HCE-T cells. Conclusions: The reduction in CMTM6 expression on the ocular surface contributes to the pathogenesis of DED. The CMTM6-NF-κB p65 signaling pathway may serve as a promising therapeutic target for DED.

Clinical and genetic profiles of chinese pediatric patients with catecholaminergic polymorphic ventricular tachycardia.

BACKGROUNDS: Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a rare but lethal cardiac ion channelopathy. Delayed diagnosis and misdiagnosis remain a matter of concern due to its rarity and insufficient recognition of this disorder, particularly in developing countries like China. AIMS AND METHODS: We reported six catecholaminergic polymorphic ventricular tachycardia (CPVT) children diagnosed in our center along with a comprehensive review of Chinese pediatric CPVT patients reported in domestic and overseas literature between January 2013 and December 2021 to provide an essential reference for physicians to deepen their understanding of pediatric CPVT. RESULTS: A total of 95 children with CPVT, including our six patients from 21 medical centers were identified. The median age of symptom onset is 8.7 ± 3.0 years. Diagnosis occurred at a median age of 12.9 ± 6.8 years with a delay of 4.3 ± 6.6 years. Selective beta-blockers (Metoprolol and Bisoprolol) were prescribed for 38 patients (56.7%) and 29 (43.3%) patients received non-selective beta-blocker (Propranolol and Nadolol) treatment. Six patients accepted LCSD and seven received ICD implantation at the subsequent therapy. A total of 13 patients died during the disease course. Of the 67 patients with positive gene test results, variants in RYR2 were 47 (70.1%), CASQ2 were 11 (16.4%), and RYR2 accompanied SCN5A were 7 (10.4%). Patients with CASQ2 gene mutations presented with younger symptom onset age, higher positive family history rate and better prognosis than those with RYR2 mutations. CONCLUSION: Chinese pediatric patients with CPVT had a poorer prognosis than other cohorts, probably due to delayed/missed diagnosis, non-standard usage of beta-blockers, unavailability of flecainide, and a lower rate of LCSD and ICD implantation.

Assessing the efficacy of four methods established by four parameters in ICL size selection and relevant influencing factors: a prospective cohort study.

PURPOSE: To compare the efficacy and relevant influencing factors of four ICL size selection methods established by four different parameters. METHODS: This prospective study included 60 patients (120 eyes) who underwent bilateral ICL implantation. Patients were equally divided into four groups, and each group used the Parkhurst nomogram based on sulcus-to-sulcus (STS), the manufacturer's Online Calculation & Ordering System (OCOS) nomogram based on white-to-white (WTW), the KS formula based on angle-to-angle (ATA) and the NK formula based on anterior chamber width (ACW) to determine the ICL size. Recorded the vault one month after operation and compared the consistency between STS and WTW, ATA and ACW and their effects on the vault of different groups. RESULTS: The Parkhurst nomogram, OCOS nomogram, KS formula and NK formula determined 86.7%, 70.0%, 83.3% and 66.7% of properly sized ICL, respectively. STS and ATA were correlated (P < 0.05). The mean difference between the STS and WTW, ATA and ACW was -0.37 ± 0.62 mm, -0.42 ± 0.53 mm and -0.44 ± 0.52 mm, respectively. The vault in the OCOS group was negatively correlated with △STS-WTW, and the vault in the NK group was negatively correlated with △STS-WTW, △STS-ATA and △STA-ACW. The vault in the Parkhurst group and KS group was not affected by anterior segment biometry variables. CONCLUSION: ATA can be served as an alternative parameter to STS, and STS-based Parkhurst nomogram and ATA-based KS formula determined the most appropriate ICL size. When using OCOS nomogram and NK formula to select ICL size, postoperative abnormal vault was associated with a larger difference between STS and other anterior segment parameters.

Clinical Characteristics and Tear Film Biomarkers in Patients With Chronic Dry Eye Disease After Femtosecond Laser-Assisted Laser in Situ Keratomileusis.

PURPOSE: To investigate clinical characteristics and tear film biomarkers of patients with chronic dry eye disease (DED) following femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK). METHODS: Patients were divided into the chronic DED after FS-LASIK (n = 36), DED without FS-LASIK (n = 39), and normal control (without FS-LASIK; n = 34) groups. Dry eye, pain, and psychological-related symptoms were evaluated using the Ocular Surface Disease Index (OSDI), Numerical Rating Scale (NRS), Neuropathic Pain Symptom Inventory Modified for the Eye (NPSI-Eye), and Hamilton Anxiety Rating Scale (HAMA) questionnaires. Ocular surface parameters, tear cytokines, and neuropeptide concentrations were evaluated with specific tests. RESULTS: The DED after FS-LASIK group showed higher corneal fluorescein staining scores, but lower OSDI and NPSI-Eye scores than the DED without FS-LASIK group (all P < .05). Corneal sensitivity and nerve density decreased in the DED after FS-LASIK group (all P < .01). Granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-17A, IL-23, alpha-melanocyte stimulating hormone (α-MSH), oxytocin, and substance P levels were highest in the DED after FS-LASIK group, followed by the DED without FS-LASIK and normal control groups (all P < .05). Interferon-γ and neurotensin levels were only significantly higher in the DED after FS-LASIK group (all P < .05). CONCLUSIONS: Patients with chronic DED after FS-LASIK showed milder ocular symptoms, greater epithelial damage, and higher levels of tear inflammatory cytokines and neuropeptides than patients with DED without FS-LASIK, indicating that the nervous and immune systems may play significant roles in FS-LASIK-related chronic DED development. [J Refract Surg. 2023;39(8):556-563.].

Evaluation of the Performance of Two Nomograms and Four Vault Prediction Formulas for Implantable Collamer Lens Size Selection.

PURPOSE: To evaluate the performance of different nomograms and vault prediction formulas in predicting the optimal Implantable Collamer Lens (ICL; STAAR Surgical) size and vault. METHODS: This retrospective study included 108 participants (214 eyes) who underwent ICL implantation. The efficacy of the manufacturer's nomogram, Parkhurst nomogram, NK formula, KS formula, Zhu formula, and ZZ formula was assessed by comparing the indicated ICL sizes to the postoperative vault measurements. Additionally, Bland-Altman plots and the Friedman test were used to assess the agreement and absolute error between the actual vault and predicted vault. RESULTS: The proportions of the ideal ICL category recommended by the manufacturer's nomogram, Parkhurst nomogram, NK formula, KS formula, Zhu formula, and ZZ formula were 50.5%, 45.3%, 46.7%, 42.5%, 50.0%, and 28.5%, respectively. The mean differences between the actual vault and the predicted vault using the NK, KS, Zhu, and ZZ formulas were 144.1 ± 261.1, -19.3 ± 179.6, 70.8 ± 284.2, and 182.6 ± 361.5 µm, respectively. The predicted ICL vault tended to overestimate the actual ICL vault, particularly when choosing a larger ICL size. The KS formula shows the smallest deviation in prediction error and is least affected by variation in ICL size. CONCLUSIONS: White-to-white distance from the Pentacam (Oculus Optikgeräte GmbH) coupled with the manufacturer's nomogram performed well for ICL size selection. Four vault prediction formulas tended to overestimate the actual ICL vault, particularly when selecting a larger ICL size. The KS formula appeared to have the least bias of the formulas. Simultaneously, vault prediction formulas need to be modified according to the ICL size. [J Refract Surg. 2023;39(7):456-461.].

Impaired Cardiomyocyte Maturation Leading to DCM: A Case Report and Literature Review.

Background: The maturation of cardiomyocytes is a rapidly evolving area of research within the field of cardiovascular medicine. Understanding the molecular mechanisms underlying cardiomyocyte maturation is essential to advancing our knowledge of the underlying causes of cardiovascular disease. Impaired maturation can lead to the development of cardiomyopathy, particularly dilated cardiomyopathy (DCM). Recent studies have confirmed the involvement of the ACTN2 and RYR2 genes in the maturation process, facilitating the functional maturation of the sarcomere and calcium handling. Defective sarcomere and electrophysiological maturation have been linked to severe forms of cardiomyopathy. This report presents a rare case of DCM with myocardial non-compaction, probably resulting from allelic collapse of both the ACTN2 and RYR2 genes. Case Presentation: The proband in this case was a four-year-old male child who presented with a recurrent and aggressive reduction in activity tolerance, decreased ingestion volume, and profuse sweating. Electrocardiography revealed significant ST-T segment depression (II, III, aVF V3-V6 ST segment depression >0.05 mV with inverted T-waves). Echocardiography showed an enlarged left ventricle and marked myocardial non-compaction. Cardiac magnetic resonance imaging revealed increased left ventricular trabeculae, an enlarged left ventricle, and a reduced ejection fraction. Whole exome sequencing revealed a restricted genomic depletion in the 1q43 region (chr1:236,686,454-237,833,988/Hg38), encompassing the coding genes ACTN2, MTR, and RYR2. The identified variant resulted in heterozygous variations in these three genes, with the ACTN2 g.236,686,454-236,764,631_del and RYR2 g.237,402,134-237,833,988_del variants being the dominant contributors to the induction of cardiomyopathy. The patient was finally diagnosed with DCM and left ventricular myocardial non-compaction. Conclusions: This study reports a rare case of DCM with myocardial non-compaction caused by the allelic collapse of the ACTN2 and RYR2 genes. This case provides the first human validation of the critical role of cardiomyocyte maturation in maintaining cardiac function and stability and confirms the key findings of previous experimental research conducted by our group. This report emphasizes the connection between genes involved in regulating the maturation of cardiomyocytes and the development of cardiomyopathy.

Misdiagnosed myocarditis in arrhythmogenic cardiomyopathy induced by a homozygous variant of DSG2: a case report.

Background: Arrhythmogenic cardiomyopathy (ACM) is an inherited cardiomyopathy that is rarely diagnosed in infants or young children. However, some significant homozygous or compound heterozygous variants contribute to more severe clinical manifestations. In addition, inflammation of the myocardium and ventricular arrhythmia might lead to misdiagnosis with myocarditis. Here, we describe an 8-year-old patient who had been misdiagnosed with myocarditis. Timely genetic sequencing helped to identify this case as ACM induced by a homozygous variant of DSG2. Case presentation: The proband of this case was an 8-year-old boy who initially presented with chest pain with an increased level of cardiac Troponin I. In addition, the electrocardiogram revealed multiple premature ventricular beats. Cardiac magnetic resonance revealed myocardial edema in the lateral ventricular wall and apex, indicating localized injuries of the myocardium. The patient was primarily suspected to have acute coronary syndrome or viral myocarditis. Whole-exome sequencing confirmed that the proband had a homozygous variation, c.1592T > G, of the DSG2 gene. This mutation site was regulated by DNA modification, which induced amino acid sequence changes, protein structure effects, and splice site changes. According to MutationTaster and PolyPhen-2 analyses, the variant was considered a disease-causing mutation. Next, we used SWISS-MODEL to illustrate the mutation site of p.F531C. The ensemble variance of p.F531C indicated the free energy changes after the amino acid change. Conclusion: In summary, we reported a rare pediatric case initially presenting as myocarditis that transitioned into ACM during follow-up. A homozygous genetic variant of DSG2 was inherited in the proband. This study expanded the clinical feature spectrum of DSG2-associated ACM at an early age. Additionally, the presentation of this case emphasized the difference between homozygous and heterozygous variants of desmosomal genes in disease progression. Genetic sequencing screening could be helpful in distinguishing unexplained myocarditis in children.

Evaluation of biometric indicators of anterior segment parameters after ICL implantation by swept-source optical coherence tomography.

BACKGROUND: To evaluate anterior segment structural alterations after implantable collamer lens (ICL) implantation in myopic patients using swept-source quantitative optical coherence tomography (SS-OCT). METHODS: This prospective study included 47 eyes in 24 patients with preoperative spherical equivalent ≥ -3.00 D. Patients underwent ICL implantation at Department of Ophthalmology, Peking University Third Hospital, from May 2021 to December 2022. SS-OCT was used to measure anterior chamber width (ACW), angle opening distance (AOD), angle recess area (ARA), trabecular-iris area (TISA), trabecular-iris angle (TIA), iridotrabecular contact (ITC) area, and ITC Index before ICL implantation surgery and at 1 month follow-up. The correlations among the ITC index, vault, and angle parameters were analysed. Receiver operating characteristic (ROC) analysis was used to explore the ability of the vault to identify eyes with suspected angle-closure. RESULTS: At one month following ICL implantation, the ITC area was 0.396 ± 0.37 mm2, and the ITC index is 8.143 ± 5.439%. All angle parameters, except ACW, showed a statistically significant reduction on SS-OCT (P < 0.05). Mean AOD500, AOD750, ARA500, ARA750, TISA500, TISA750, TIA500, and TIA750 values at one month postoperatively decreased by 60.0%, 60.4%, 58.1%, 59.2%, 57.3%, 58.7%, 48.8%, and 50.7%, respectively. The vault was positively correlated with the ITC index and percent change in anterior chamber angle parameters. A vault of > 0.659 mm was found to be optimal for angle-closure suspect with a sensitivity of 85.2% and a specificity of 53.9%. CONCLUSIONS: Anterior chamber angle parameters decreased one month after ICL implantation, and their percentage changes and ITC index correlated with the vault. When the vault is larger than 0.659 mm, it is necessary to be alert to possible closed angle suspicion.

The impact of Water Ecological Civilization City Pilot on urban green innovation: the case of China.

As an essential part of ecological civilization, water ecological civilization has significant influence on the green and sustainable development of cities. Under the background of China's Water Ecological Civilization City Pilot (WECCP), based on data from 275 cities in China from 2007 to 2019 by using the difference-in-differences (DID) model, we empirically analyzed the influence of the WECCP establishment on urban green innovation and explored the impact mechanism in depth using a mediating effect model, which aimed to verify whether the "Porter hypothesis" holds true in China. The result indicated that the WECCP had made a remarkable contribution to enhancing urban green innovation in the pilot cities. Further research found that the input mechanism played an important mediating role. In addition, the heterogeneity test indicated that cities in the central region, at low administrative levels, and in the first batch of pilots gained more from the policy establishment. This paper has theoretical implications for understanding the derived innovation benefits of the environmental policy, practical implications for identifying new drivers of urban innovation, and provides related experience for the country to further promote and expand water ecological civilization construction and useful policy inspiration for other developing countries to formulate ecological and environmental policies.

Case report: cerebral venous sinus thrombosis and pulmonary embolism as the initial presentation in a child with asymptomatic primary nephrotic syndrome.

Background: Cerebral venous sinus thrombosis (CVST) is rare, but potentially life-threatening. The clinical course definitely become more unpredictable and fatal in patients complicated by pulmonary embolism (PE). Nephrotic syndrome (NS) is an uncommon etiology of CVST. Concurrence of CVST and PE at the initial onset of NS is extremely unusual and rarely reported. Considering that edema might be absent in NS individuals, thromboembolic events probably become unrecognized, thereby causing a missed or delayed diagnosis and poor outcome. Herein, we described an extraordinary case of an adolescent boy presenting with both CVST and PE initially just within 5 days of disease onset, who was ultimately diagnosed with asymptomatic NS, aiming to emphasize a high index of suspicion of these diseases in patients with conditions of hypercoagulability. Case presentation: A 13-year-old male child presented acutely with dizziness, fever and dyspnea, with signs of shock but undetected edema. Initial laboratory investigations revealed hypoalbuminemia, typical images of pneumonia, and normal radiographic findings on non-enhanced computed tomography of head. Despite evidence of hypoalbuminemia and neurological symptoms, the child was still misdiagnosed as pneumonia. His dyspnea and period of headache deteriorated even if hemodynamic stability and undetected fever after initial therapy. The delayed urinalysis and 24-h urine examination both showed massive proteinuria. A computed tomography angiography of chest along with cranial magnetic resonance imaging/magnetic resonance venography were subsequently performed, consistent with the imaging features of PE and CVST, respectively. The diagnosis of asymptomatic primary NS complicated by PE and CVST was ultimately confirmed. The patient received corticosteroids and antithrombotic therapy with satisfactory results. Conclusion: A persistent clinical suspicion of CVST should be borne in mind in patients with a sudden, new or worsening headache, specifically among those with prothrombotic conditions. NS should always be considered in the differential diagnosis of risk factors for CVST, even in absence of edema. Since CVST and PE can be present simultaneously at extraordinary early-onset of NS, early radiological diagnosis is clinically substantial to proper management and satisfactory long-term outcomes.

Comparison of mucin levels at the ocular surface of visual display terminal users with and without dry eye disease.

BACKGROUND: The long-term use of visual display terminals (VDT) is linked to an increased risk of dry eye disease (DED). Numerous studies have indicated that ocular mucins play a vital role in the pathogenesis of DED. Therefore, we aimed to evaluate (1) whether mRNA levels of membrane-associated mucins (MAMs), including MUC1, MUC4, MUC16, and MUC20, as well as MUC5AC are altered in conjunctival cells of VDT users with and without DED and (2) the relationship between mucin levels and subjective and objective tests of DED in VDT users. METHODS: Seventy-nine VDT users were enrolled and divided into DED (n = 53) and control (n = 26) groups. All participants were evaluated for parameters of DED using the Ocular Surface Disease Index (OSDI) questionnaire, tear breakup time (TBUT), corneal fluorescein staining (CFS), lissamine green (LG) staining, and tear meniscus height (TMH). Based on the conjunctival impression cytology (CIC) method, differences in MUC1, MUC4, MUC16, MUC20, and MUC5AC mRNA expression levels were observed between the DED and control groups, and between symptomatic and asymptomatic participants. RESULTS: The DED group showed significantly decreased MUC1, MUC16, and MUC20 expressions (all P < 0.05) compared to the control group. In addition, these mucin levels were lower in subjects with frequent ocular symptoms (foreign body sensation, blurred vision and painful or sore eyes) than in asymptomatic participants (all P < 0.05). Correlation analysis revealed that MUC1, MUC16, and MUC20 levels in VDT users were positively correlated with TBUT or TMH, or both. However, no significant relationship was found between MUC4 and MUC5AC levels and the DED parameters. CONCLUSION: VDT users with an increased frequency of ocular discomfort or a diagnosis of DED had a decreased MUC1, MUC16 and MUC20 mRNA expression in their conjunctival cells. MAMs deficiency in the conjunctival epithelium may be one of the mechanisms leading to tear film instability and DED in VDT users.

Elevated Neuropeptides in Dry Eye Disease and Their Clinical Correlations.

PURPOSE: The goal of this study was to assess neuropeptide levels in patients with dry eye disease (DED) and investigate their correlations with clinical characteristics. METHODS: This study included 38 eyes of 38 patients diagnosed with DED (DED group) and 38 eyes of 38 healthy volunteers without DED (control group). Ocular surface evaluation was performed. The severity of dry eye symptoms and signs in the DED group was graded. Neuropeptides [substance P (SP), alpha-melanocyte-stimulating hormone (α-MSH), ß-endorphin, neurotensin, and oxytocin] and inflammatory cytokines levels were measured in basal tears. The link between neuropeptides and clinical parameters was investigated using Spearman rank correlation. RESULTS: Overall, 76.3% of patients in the DED group showed dry eye symptoms and signs that were inconsistent in severity. Compared with the control group, the DED group showed higher levels of SP, α-MSH, and oxytocin in tears (P = 0.012, P = 0.030, and P = 0.006, respectively), but similar levels of ß-endorphin and neurotensin (P = 0.269 and P = 0.052). The levels of SP, α-MSH, and oxytocin were elevated in DED patients with higher grading of symptoms than clinical signs (all P < 0.05). SP, α-MSH, and oxytocin levels in tears were positively correlated with Ocular Surface Disease Index scores, frequency of sensitivity to light, and frequency of blurred vision (all P < 0.05). CONCLUSIONS: The increased tear levels of SP, α-MSH, and oxytocin may be linked to ocular discomfort in DED. Neuropeptides may play a key role in the development of DED, especially in DED patients with more severe symptoms than clinical signs.

Mapping Resident Immune Cells in the Murine Ocular Surface and Lacrimal Gland by Flow Cytometry.

BACKGROUND: The ocular surface and lacrimal gland have a frontline position in mucosal immunology. However, there have been few updates to the immune cell atlas of these tissues in recent years. PURPOSE: To map the immune cells in murine ocular surface tissues and lacrimal gland. METHODS: Central and peripheral corneas, conjunctiva, and lacrimal gland were dissociated into single cell suspensions, followed by flow cytometry. Discrepancy of immune cells between the central and peripheral corneas was compared. In the conjunctiva and lacrimal gland, myeloid cells were clustered by tSNE and FlowSOM based on the expression of F4/80, Ly6C, Ly6G, and MHC II. ILCs, type 1 immune cells, and type 3 immune cells were analyzed. RESULTS: The number of immune cells in peripheral corneas was about 16 folds of that in central corneas. B cells accounted for 8.74% of immune cells in murine peripheral corneas. In the conjunctiva and lacrimal gland, most myeloid cells tended out to be monocytes, macrophages, and classical dendritic cells (cDCs). ILC3 were 6.28% and 3.63% of ILCs in the conjunctiva and lacrimal gland, respectively. Th1, Tc1, and NK cells were predominant type 1 immune cells. γδ T17 cells and ILC3 outnumbered Th17 cells among type 3 T cells. CONCLUSION: B cells resident in murine corneas were reported for the first time. Additionally, we proposed a strategy of clustering myeloid cells to better understand their heterogeneity in the conjunctiva and lacrimal gland based on tSNE and FlowSOM. Furthermore, we identified the ILC3 in the conjunctiva and lacrimal gland for the first time. Compositions of type 1 and type 3 immune cells were summarized. Our study provides a fundamental reference and novel insights for ocular surface immune homeostasis and diseases.

Characterization of the Retinal Microvasculature and FAZ Changes in Ischemic Stroke and Its Different Types.

Purpose: This study aimed to assess morphological changes in the retinal microvasculature and foveal avascular zone (FAZ) in patients with ischemic stroke and its different subtypes. Methods: Thirty-three patients with ischemic stroke (14 with nonlacunar infarction and 19 with lacunar infarction) and 27 control participants were enrolled in this study. Based on optical coherence tomography angiography (OCTA), three vascular parameters, including vascular area density, vascular fractal dimension (VFD), and vascular orientation distribution (VOD), and four FAZ-related parameters, including FAZ area, FAZ axis ratio (FAR), FAZ circularity (FC), and FAZ roundness, in the superficial capillary plexus (SCP) and deep capillary plexus (DCP) were extracted and analyzed. Results: Logistic regression results showed that worse best-corrected visual acuity (odds ratio [OR], 0.21), higher FAR (OR, 2.77) and lower FC (OR, 0.36) of the DCP were associated with ischemic stroke. Furthermore, lower VOD of the SCP was significantly associated with lacunar infarction compared with nonlacunar infarction. Conclusions: Our study shows that the FAR and FC of the DCP may be potential biomarkers of ischemic stroke. Moreover, we demonstrated that OCT showed specific damage patterns in retinal microvascular and macular morphology in different subtypes of ischemic stroke. Translational Relevance: This work lays the foundation for the pathophysiological characteristics of cerebrovascular diseases assisted by retinal imaging and artificial intelligence.

Progress of Bulbar Conjunctival Microcirculation Alterations in the Diagnosis of Ocular Diseases.

Bulbar conjunctival microcirculation is a microvascular system distributed in the translucent bulbar conjunctiva near the corneal limbus. Multiple ocular diseases lead to bulbar conjunctival microcirculation alterations, which means that bulbar conjunctival microcirculation alterations would be potential screening and diagnostic indicators for these ocular diseases. In recent years, with the emergence and application of a variety of noninvasive observation devices for bulbar conjunctiva microcirculation and new image processing technologies, studies that explored the potential of bulbar conjunctival microcirculation alterations in the diagnosis of ocular diseases have been emerging. However, the potential of bulbar conjunctival microcirculation alterations as indicators for ocular diseases has not been exploited to full advantage. The observation devices, image processing methods, and algorithms are not unified. And large-scale research is needed to concrete bulbar conjunctival microcirculation alterations as indicators for ocular diseases. In this paper, we provide an update on the progress of bulbar conjunctival microcirculation alterations in the diagnosis of ocular diseases in recent five years (from January 2017 to March 2022). Relevant ocular diseases include contact lens wearing, dry eye, conjunctival malignant melanoma, conjunctival nevus, and diabetic retinopathy.