BackgroundRetinoblastoma (RB) is the most common intraocular malignant tumor in children. It not only seriously threatens patients’ vision but also endangers their lives if not treated in time. Our objective is to analyze research trends in the RB field and compare contributions from different countries, institutions and authors.MethodsWe extracted all RB-related publications published from 1980 to 2023 from the Web of Science database and applied VOSviewer, R software, Bibliometrix software, Origin 2024 and CiteSpace to review the publication data, analyze the publication trends, and visualize the relevant data. In this study, the research papers on RB published in the past 44 years were classified by year, country/region, institution/university, journal, author and keywords to reveal the research hotspots and development trends in this field.ResultsA total of 4156 papers on RB were identified from 1980 to 2023. In 1980, only 13 papers were published, yet by 2023, 237 papers had been published. These publications were contributed by 351 research institutes from 68 countries/regions. The United States ranked first with 1662 papers, accounting for 39.99% of the total number of publications on RB research. A total of 539 RB research papers were published in China, ranking second. India, Canada and Germany ranked third, fourth and fifth, with 377, 277 and 221 publications, respectively. Thomas Jefferson University published the most research papers on RB, with 166 published papers, accounting for 3.99% of all publications. The top three journals contributing to this field were Invest Ophth Vis Sci, the British Journal of Ophthalmology and Ophthalmology. Liquid biopsy, intra-arterial chemotherapy and intravitreal chemotherapy are the most frequently used keywords in the field.ConclusionOver the past 44 years, the United States, China, India, Canada and Germany have led the field of research on RB. Many renowned research institutions and ophthalmologists have made important contributions to RB research and will continue to lead this research direction. Liquid biopsy, intra-arterial chemotherapy and intravitreal chemotherapy are potential hotspots for RB research in the future.

Extended Boundary Condition Method (EBCM) is a highly efficient technique utilized for the analysis of electromagnetic (EM) scattering from periodic or quasi-periodic surfaces. This article investigates the validity of EBCM when it comes to solving the EM scattering from random rough surfaces. Two advanced EBCM techniques were evaluated on three distinct types of rough surfaces. Simulations were performed to ascertain the valid range of EBCM to rough surfaces with varied attributes. An indicator for ill-conditioned matrices is introduced, and the conservation of energy is scrutinized under a variety of conditions. Ultimately, it has been empirically verified that the validity of the EBCM is confined to surfaces with specific characteristics.

Purpose: This study aimed to explore the effects of elevated KDM4D expression and potential therapeutic effects of Lycium barbarum polysaccharide (LBP) on pterygium.Methods: The expression levels of KDM4D in the primary pterygium (n = 29) and normal conjunctiva (n = 14) were detected by immunohistochemistry. The effects of KDM4D on pterygium fibroblasts were detected by the CCK-8 assay, liquid chromatography-mass spectrometry assay, flow cytometry, and scratch wound healing assay. The relative expression of KDM4D in pterygium fibroblasts stimulated by interleukin (IL)-1 beta, IL-6, IL-8, and LBP was detected by quantitative real-time PCR and Western blot. The effects of LBP on pterygium fibroblasts were detected using flow cytometry and scratch wound healing assays.Results: The expression level of KDM4D in pterygium was higher than that in normal conjunctiva. KDM4D increased the cell viability of pterygium fibroblasts. The differentially expressed genes identified in the LM-MS assay enriched in "actin filament organization" and "apoptosis." KDM4D promoted migration and inhibited apoptosis of pterygium fibroblasts in vitro. Inflammatory cytokines, including IL-1 beta, IL-6, and IL-8, enhanced the expression of KDM4D in pterygium fibroblasts. LBP inhibited the expression of KDM4D in pterygium fibroblasts and decreased their cell viability. Moreover, LBP attenuated the KDM4D effects on migration and apoptosis of pterygium fibroblasts.Conclusions: Elevated KDM4D expression is a risk factor for pterygium formation. LBP inhibits the expression of KDM4D in pterygium fibroblasts and may be a potential drug for delaying pterygium development.

Purpose: Corneal injury (CI) resulting in corneal opacity remains a clinical challenge. Exosomes (Exos) derived from bone marrow mesenchymal stem cells (BMSCs) have been proven effective in repairing various tissue injuries and are also considered excellent drug carriers due to their biological properties. Recently, microRNA-29b (miR-29b) was found to play an important role in the autophagy regulation which correlates with cell inflammation and fibrosis. However, the effects of miR-29b and autophagy on CI remain unclear. To find better treatments for CI, we used Exos to carry miR-29b and investigated its effects in the treatment of CI. Methods: BMSCs were transfected with miR-29b-3p agomir/antagomir and negative controls (NCs) to obtain Exos-29b-ago, Exos-29b-anta, and Exos-NC. C57BL/6J mice that underwent CI surgeries were treated with Exos-29b-ago, Exos-29b-anta, Exos-NC, or PBS. The autophagy, inflammation, and fibrosis of the cornea were estimated by slit-lamp, hematoxylin and eosin (H&E) staining, immunofluorescence, RT‒qPCR, and Western blot. The effects of miR-29b-3p on autophagy and inflammation in immortalized human corneal epithelial cells (iHCECs) were also investigated. Results: Compared to PBS, Exos-29b-ago, Exos-29b-anta, and Exos-NC all could ameliorate corneal inflammation and fibrosis. However, Exos-29b-ago, which accumulated a large amount of miR-29b-3p, exerted excellent potency via autophagy activation by inhibiting the PI3K/AKT/mTOR pathway and further inhibited corneal inflammation via the mTOR/NF-κB/IL-1β pathway. After Exos-29b-ago treatment, the expressions of collagen type III, α-smooth muscle actin, fibronectin, and vimentin were significantly decreased than in other groups. In addition, overexpression of miR-29b-3p prevented iHCECs from autophagy impairment and inflammatory injury. Conclusions: Exos from BMSCs carrying miR-29b-3p can significantly improve the therapeutic effect on CI via activating autophagy and further inhibiting corneal inflammation and fibrosis.

PDF file 373K, Supplementary Figure 1: Staining of additional Ewing sarcoma cell lines. Supplementary Figure 2: MRI detection of experimental metastases

Deposition of misfolded α-synuclein (α-Syn) aggregates in the human brain is one of the major hallmarks of synucleinopathies. Positron Emission Tomography imaging (PET) of α-synuclein in Parkinson’s disease (PD) patients is highly desirable but remains elusive. An extensive PET radioligand discovery campaign was undertaken. Here, we present the in-vitro and preclinical in-vivo characterization of [ 3 H]- and [ 11 C]Compound S, representing an example of a promising PET radioligand for α-Syn. Competition binding studies and autoradiographic studies were performed in cortexes of PD patients, healthy control, and Alzheimer’s disease (AD) patients brain tissue, and mid-brain region of the aged A30P mouse using [ 3 H]S. Brain PET experiments using [ 11 C]S were carried out in a healthy non-human primate (NHP) and the aged homozygote A30P mice. Compound S is a sub-nanomolar affinity ligand to human and A30P mouse α-Syn (Kd = 0.15 nM and 0.5 nM resp.). Lower affinity was observed in AD and healthy control brain tissues. [ 3 H]S showed displaceable binding in PD brain cortex and A30P mid-brain and brainstem, and little binding in AD and healthy control. In the NHP, [ 11 C]S crosses the brain-blood barrier and the uptake phase of [ 11 C]S is followed by a washout. [ 11 C]S PET studies in A30P mice showed higher uptake in the mid brain and brainstem regions, with standard uptake value ratios in those regions higher than 2.0. Compound S is potent for α-Syn in vitro, and PET imaging in the A30P mice showed an elevated signal in regions known to accumulate α-Syn. This dataset suggests that the Compound S has promise towards imaging α-Syn in PD patients .

Purpose: To identify the molecular background of eyelid sebaceous gland carcinomas (SCs), we conducted the integrated whole-exome sequencing and transcriptome sequencing for eyelid SCs in this study. Methods: The genetic alterations were studied by whole-exome sequencing, and the messenger RNA expression was studied using Oxford Nanopore Technologies (ONT) in five paired fresh eyelid SC tissues and adjacent normal tissues. Integrated analysis of exome and transcriptomic information was conducted for filtering candidate driver genes. Protein–protein interaction (PPI) network of filtered candidate genes was analyzed by STRING. The protein expression was verified by immunohistochemistry in 29 eyelid SCs and 17 compared normal sebaceous gland tissues. Results: The average numbers of pathogenic somatic single-nucleotide variants (SNVs) and indels in eyelid SCs were 75 and 28, respectively. Tumor protein p53 (TP53), zinc finger protein 750 (ZNF750), filaggrin 2 (FLG2), valosin-containing protein (VCP), and zinc finger protein 717 (ZNF717) were recurrent mutated genes. A mean of 844 differentially expressed genes (DEGs) were upregulated, and 1401 DEGs were downregulated in SC samples. The intersection of DEG-based pathways and mutation-based pathways was mainly involved in microbial infection and inflammation, immunodeficiency, cancer, lipid metabolism, and the other pathways. The intersection of DEGs and mutated genes consisted of 55 genes, of which 15 genes formed a PPI network with 4 clusters. The PPI cluster composed of scavenger receptor class B member 1 (SCARB1), peroxisome proliferator–activated receptor γ (PPARG), peroxisome proliferator–activated receptor γ coactivator 1α (PPARGC1A) was involved in cholesterol metabolism. The expression of SCARB1 protein was found to be increased, whereas that of PPARG protein was decreased in eyelid SCs compared to that in the normal sebaceous glands. Conclusions: Increased SCARB1 and decreased PPARG indicated that dysregulation of cholesterol metabolism might be involved in carcinogenesis of eyelid SCs. Translational Relevance: The malfunction in cholesterol metabolism might advance our knowledge of the carcinogenesis of eyelid SCs.

PDF file 17K, Supplementary Figure 1: Staining of additional Ewing sarcoma cell lines. All Ewing sarcoma cell lines tested stain equivalently positive for DN16. In all panels, unstained and isotype matched IgG are controls. Supplementary Figure 2: MRI detection of experimental metastases. Experimental liver metastases were induced by IV injection of TC32 cells. Imaging with a 7T MRI demonstrates 1-2 mm metastatic lesions in the liver (white arrows) -

Inflammation is a key factor in the pathogenesis of dry eye disease (DED). We aimed to investigate the role of microRNA-146a (miR-146a) in regulating corneal inflammation in a mouse model of benzalkonium chloride (BAC)-induced dry eye and the TNF-α-induced NF-κB signaling pathway in human corneal epithelial cells (HCECs). A mouse model of dry eye was established by administering with BAC to BALB/c mice, and the expression of TNF-α, IL-1β, IL-6, IL-8, cyclooxygenase 2 (COX2), interleukin-1 receptor-associated kinase 1 (IRAK1) and TNF receptor-associated factor 6 (TRAF6) in the corneas of dry eye model mice was significantly increased; this was accompanied by the upregulation of miR-146a and activation of the NF-κB pathway. In vitro, TNF-α induced miR-146a expression in HCECs, while the NF-κB inhibitor SC-514 reduced the expression of miR-146a. Overexpression of miR-146a decreased the expression of IRAK1 and TRAF6, which have been identified as targets of miR-146a. Furthermore, overexpression of miR-146a suppressed NF-κB p65 translocation from the cytoplasm to the nucleus. Moreover, overexpression of miR-146a attenuated the TNF-α-induced expression of IL-6, IL-8, COX2 and intercellular adhesion molecule 1 (ICAM1), while inhibition of miR-146a exerted the opposite effect. Our results suggest that miR-146a mediates the inflammatory response in DED. MiR-146a negatively regulates inflammation in HCECs through the IRAK1/TRAF6/NF-κB pathway, and this may serve as a potential therapeutic approach for the treatment of DED.

Positron emission tomography (PET) ligands play an important role in the development of therapeutics by serving as target engagement or pharmacodynamic biomarkers. Here, we describe the discovery and translation of the PET tracer [11C]MK-6884 from rhesus monkeys to patients with Alzheimer’s disease (AD). [3H]MK-6884/[11C]MK-6884 binds with high binding affinity and good selectivity to an allosteric site on M4 muscarinic cholinergic receptors (M4Rs) in vitro and shows a regional distribution in the brain consistent with M4R localization in vivo. The tracer demonstrates target engagement of positive allosteric modulators of the M4R (M4 PAMs) through competitive binding interactions. [11C]MK-6884 binding is enhanced in vitro by the orthosteric M4R agonist carbachol and indirectly in vivo by the acetylcholinesterase inhibitor donepezil in rhesus monkeys and healthy volunteers, consistent with its pharmacology as a highly cooperative M4 PAM. PET imaging of [11C]MK-6884 in patients with AD identified substantial regional differences quantified as nondisplaceable binding potential (BPND) of [11C]MK-6884. These results suggest that [11C]MK-6884 is a useful target engagement biomarker for M4 PAMs but may also act as a sensitive probe of neuropathological changes in the brains of patients with AD.

To report on the first group of AD patients studied with a new M4 muscarinic cholinergic receptor PET tracer, [11C]MK-6884.

Purpose Programmed cell death-1 receptor (PD-1) and its ligand (PD-L1) are the targets for immunotherapy in many cancer types. Although PD-1 blockade has therapeutic effects, the efficacy differs between patients. Factors contributing to this variability are PD-L1 expression levels and immune cells present in tumors. However, it is not well understood how PD-1 expression in the tumor microenvironment impacts immunotherapy response. Thus, imaging of PD-1-expressing immune cells is of interest. This study aims to evaluate the biodistribution of Zirconium-89 ( 89 Zr)-labeled pembrolizumab, a humanized IgG4 kappa monoclonal antibody targeting PD-1, in healthy cynomolgus monkeys as a translational model of tracking PD-1-positive immune cells. Procedures Pembrolizumab was conjugated with the tetrafluorophenol-N-succinyl desferal-Fe(III) ester (TFP-N-sucDf) and subsequently radiolabeled with 89 Zr. Four cynomolgus monkeys with no previous exposure to humanized monoclonal antibodies received tracer only or tracer co-injected with pembrolizumab intravenously over 5 min. Thereafter, a static whole-body positron emission tomography (PET) scan was acquired with 10 min per bed position on days 0, 2, 5, and 7. Image-derived standardized uptake values (SUV mean ) were quantified by region of interest (ROI) analysis. Results 89 Zr-N-sucDf-pembrolizumab was synthesized with high radiochemical purity (> 99 %) and acceptable molar activity (> 7 MBq/nmol). In animals dosed with tracer only, 89 Zr-N-sucDf-pembrolizumab distribution in lymphoid tissues such as mesenteric lymph nodes, spleen, and tonsils increased over time. Except for the liver, low radiotracer distribution was observed in all non-lymphoid tissue including the lung, muscle, brain, heart, and kidney. When a large excess of pembrolizumab was co-administered with a radiotracer, accumulation in the lymph nodes, spleen, and tonsils was reduced, suggestive of target-mediated accumulation. Conclusions 89 Zr-N-sucDf-pembrolizumab shows preferential uptake in the lymphoid tissues including the lymph nodes, spleen, and tonsils. 89 Zr-N-sucDf-pembrolizumab may be useful in tracking the distribution of a subset of immune cells in non-human primates and humans. Trial Registration ClinicalTrials.gov Identifier: NCT02760225

PurposeTo investigate the expression of extracellular high mobility group box 1 (HMGB1) and the effect of its inhibitor glycyrrhizin (GL) in corneal wound healing.MethodsWe treated C57BL/6J mice with GL or PBS before and after establishing a corneal injury model. Fluorescein staining, Ki-67 expression, haze grade, and haematoxylin/eosin (H&E) staining were used to assess treatment efficacy. The expression of HMGB1, NF-κB-p65, the NLRP3 inflammasome, IL-1β, CCL2, CXCL2, TGF-β1, α-SMA, fibronectin, and collagen III and neutrophil influx were examined by immunohistochemical staining, western blot, and RT-qPCR at various time points after corneal injury.ResultsAfter corneal injury, HMGB1 transferred from the nucleus to the cytoplasm and was passively released or actively secreted into the corneal stroma from epithelial cells and inflammatory cells; however, this increase was attenuated by GL treatment. Furthermore, GL indirectly attenuated the expression of IL-1β by directly inhibiting extracellular HMGB1 functions, which activated the NF-κB-p65/NLRP3/IL-1β signalling pathway. Moreover, application of GL alleviated the neutrophil infiltration that delays wound healing, accompanied by the downregulation of expression of the chemokines CCL2 and CXCL2. More interestingly, application of GL reduced the degree of haze grade through inactivating extracellular HMGB1 functions that induced TGF-β1 release and myofibroblast differentiation. In addition, fluorescein and H&E staining and Ki-67 levels suggest that GL promotes regeneration of corneal epithelium.ConclusionsAfter corneal injury, extracellular HMGB1 can be an essential driver to trigger a neutrophil- and cytokine-mediated inflammatory injury amplification loop. The application of GL promotes the cornea to restore transparency and integrity, which may be related to the attenuation of extracellular HMGB1 levels and function.

Cholinergic neuron loss has long been known to be a hallmark of AD. The largest class of therapeutic agents targets acetylcholinesterase in order to increase acetylcholine availability. In order to evaluate regional cholinergic tone in individual patients and measure target engagement for potential therapies, it would be helpful to image muscarinic receptors in vivo. Several imaging tracers have been developed over the years to image cholinergic receptors, but the findings with these agents have been either negative or too subtle to be of use. Recently, a M4 muscarinic cholinergic receptor PET tracer has been developed that shows good imaging characteristics. We report on the first group of AD patients to be studied with this tracer, [11C]MK-6884. Nine patients with Alzheimer's disease (MMSE=15.2±5.2), aged from 60 to 76 years old (mean=68.9±5.5 years, five women), underwent [11C]MK-6884 M4 receptor and [18F]FDG metabolism PET. Immediately after a bolus injection of approximately 370 MBq of [11C]MK-6884, images were acquired during 90 minutes. Standardized uptake value ratios (SUVRs) were calculated between 5 to 15 minutes using the cerebellum grey matter as the reference region. [18F]FDG SUVRs were calculated following standard procedures, using the cerebellum grey matter as the reference region between 40 to 60 min. As expected, [11C]MK-6884 showed good uptake in the basal ganglia, with low values in thalamus. Cortical uptake was reduced mostly in parieto-temporal association cortex, corresponding well with the clinical syndrome: patients with a predominant language problem had greater loss in the left hemisphere and the opposite was true for those with visuospatial skills impairment. Frontal lobes were also affected, but not as much as the posterior structures. There was a concordance in cortical [11C]MK-6884 and [18F]FDG PET values, indicating that the [11C]MK-6884 binding was disease specific, but also discordant areas, highlighting the potential specificity of [11C]MK-6884. The initial experience with [11C]MK-6884 suggests that this tracer may prove useful to image muscarinic receptors in AD.

OBJECTIVE:To detect pathogenic gene variants in two Chinese families with cone-rod dystrophy(CORD).METHODS:After the informed consent and comprehensive ophthalmic examinations for the patients, 3 mL peripheral blood was taken from the patients' blood vessel and DNA was extracted. The DNA was sequenced by whole-exome sequencing technology and variants were analyzed.RESULTS:Two novel compound heterozygous AIPL1 variants were detected in two patients, which were c.923T to C (p.L308P) and c.421C to T (p.Q141X) variants in Family 1, c.572T to C (p.L191P) and c.421C to T (p.Q141X) in Family 2.CONCLUSION:The results supported that AIPL1 gene variants are the main cause of the two CORD families. Whole-exome sequencing technology is a useful tool in the clinical differentiated diagnosis and genetic counseling for CORD patients.