Engineered Extracellular Vesicles to Enhance Antigen Presentation for Boosting Light-Driven Tumor Immunotherapy.

Extracellular vesicles (EVs) have emerged as potential tools for tumor-target therapy accompanied with activating anticancer immune responses by serving as an integrated platform, but usually suffered from the limited cross presentation of tumor-associated antigen by dendritic cells (DCs). Here, a straightforward engineering strategy to construct heat shock proteins 70 (HSP70) highly expressed EVs incapsulated with Te nanoparticles (Te@EVsHSP70 ) for tumor photothermal therapy triggering improved immunotherapy is proposed. Tumor cells are firstly used as bioreactors for intracellular synthesis of Te nanoparticles, and NIR irradiation is subsequently introduced to upregulate the expression of HSP70 to give engineered Te@EVsHSP70 through exocytosis. Te@EVsHSP70 exhibits excellent photothermal performance and enhanced tumor antigen capture capability, which induces significant immunogenic death of tumor cells and improves DCs maturation both in vitro and in vivo. Thus, the engineered EVs demonstrate superior antitumor efficacy through photothermal effect and following provoked antitumor immune responses. This work provides a facile method to fabricate multifunctional EVs-based drug delivery system for improving photothermal-triggered tumor immunotherapy.

Functional and Morphological Brain Alterations in Dysthyroid Optic Neuropathy: A Combined Resting-State fMRI and Voxel-Based Morphometry Study.

BACKGROUND: Increasing evidence has indicated that the entire visual pathway from retina to visual cortex may be involved in dysthyroid optic neuropathy (DON) pathological mechanisms. PURPOSE: To explore the functional and morphological brain characteristics in DON and their relationship with ophthalmologic performance. STUDY TYPE: Retrospective. POPULATION: A total of 30 DON patients, 40 thyroid-associated ophthalmopathy (TAO) without DON patients and 21 healthy-controls (HCs). FIELD STRENGTH/SEQUENCE: A 3.0 T, 3D T1-weighted spoiled gradient-recalled echo and gradient-recalled echo-planar imaging. ASSESSMENT: Functional and structural alterations in brain regions were evaluated with fractional amplitude of low-frequency fluctuations, degree centrality (DC), and gray matter volume (GMV). Clinical activity score (CAS) is assessed across patients. STATISTICAL TEST: One-way analysis of variance with post hoc two sample t-tests (GRF-corrected, voxel level: P < 0.005, cluster level: P < 0.05) and correlation analysis (significance level: P < 0.05). RESULTS: Compared to HCs, DON patients had significantly decreased DC values in the bilateral BA17 and BA18 regions. Compared to the TAO group, DON patients had decreased GMV in the left anterior cingulate cortex, left middle frontal gyrus, left lingual gyrus, left parietal gyrus, right Rolandic operculum, left supplementary motor area, and right middle temporal gyrus. In addition, GMV in the right Rolandic operculum was significantly positively correlated with CAS (correlation coefficient: r = 0.448). DATA CONCLUSION: This study showed significant morphological and functional alterations in visual cortex and morphological alterations in partial default mode network regions of DON patients, which may provide insights into the mechanism of vision loss and may facilitate the diagnosis and treatment of DON. EVIDENCE LEVEL: 3. TECHNICAL EFFICACY: Stage 3.

Preliminary Diffusion-Tensor Imaging Evidence for Trans-Synaptic Axonal Degeneration in Dysthyroid Optic Neuropathy Due to Thyroid-Associated Ophthalmopathy.

BACKGROUND: The mechanism driving dysthyroid optic neuropathy (DON) is unclear. Diffusion-tensor imaging (DTI) allows for noninvasively assessing the microstructure of the entire visual pathway and may facilitate a better understanding of the mechanism of DON. PURPOSE: To assess microstructural changes of the whole visual pathway and to investigate the potential mechanism of trans-synaptic damage（TSD） pathogenesis in DON with DTI. STUDY TYPE: Cross-sectional. POPULATION: Sixty-four patients with bilateral thyroid-associated ophthalmopathy (TAO), 30 with and 34 without DON, and 30 age- and sex-matched healthy controls (HCs). FIELD STRENGTH/SEQUENCE: 3 T/DTI (A single-shot diffusion-weighted echo-planar imaging sequence). ASSESSMENT: Differences in DTI parameters including fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) in each segment (optic nerve, tract, and radiation) of the entire visual pathway among the groups were compared. The parameters of visual evoked potentials (VEPs), visual field tests, and mean retinal nerve fiber layer (mRNFL) thickness on optical coherence tomography were also compared across patients. STATISTICAL TESTS: Student's t-test, chi-square test; ANOVA with post-hoc testing, interclass correlation coefficient, and correlation analysis. Significance level: P < 0.05. RESULTS: TAO patients with DON showed significantly reduced mRNFL thickness and abnormal VEPs. There was a tendency for gradually reduced FA and AD, and increased RD and MD from HCs, with non-DON to with DON in optic nerve and tract, statistically. For radiation, the RD and MD showed statistical increase, the AD and FA just showed numerical decrease (P = 0.119 and 0.059, respectively). For DON, the FA and MD of visual pathway segments showed correlations with abnormal VEPs. DATA CONCLUSION: DTI may be a useful tool for detecting microstructural changes in the entire visual pathway in DON. The changes in RNFL thickness and DTI parameters suggested TSD as a potential pathogenic mechanism of DON. EVIDENCE LEVEL: 4 Technical Efficacy: Stage 5.

Aberrant spontaneous brain activity in patients with thyroid-associated ophthalmopathy with and without optic neuropathy: a resting-state functional MRI study.

OBJECTIVES: To investigate the brain functional alterations in dysthyroid optic neuropathy (DON) by evaluating spontaneous neural activity, using functional magnetic resonance imaging (fMRI) with regional homogeneity (ReHo), and its relationship with ophthalmologic performance. METHODS: Forty-seven patients with thyroid-associated ophthalmopathy (TAO; 20 with DON, 27 with non-DON) and 33 age-, sex-, and education-matched healthy controls (HCs) underwent fMRI. ReHo values were compared using one-way analysis of variance (ANOVA) with post hoc pairwise comparisons (voxel-level p < 0.01, Gaussian random field correction, cluster-level p < 0.05). Correlations between ReHo values and ophthalmological metrics were assessed for DONs, with Bonferroni correction for multiple comparisons (p < 0.004). ROC curves were applied to evaluate the diagnostic performance of ReHo metrics. RESULTS: ReHo values were significantly lower in the left insula and right superior temporal gyrus, and higher in the left posterior cingulate cortex (LPCC), of DON than of non-DON patients. ReHo values were also significantly lower in the right middle temporal, left insula, and left precentral gyrus in DON than in HCs. Meanwhile, ReHo values were higher in LPCC in non-DON than in HCs. ReHo values correlated with ophthalmic examinations to varying degrees in DON. For distinguishing DON, the ReHo values in LPCC showed optimal individually (AUC = 0.843), the combination of the ReHo in both the left insula and LPCC performed better (AUC = 0.915). CONCLUSION: Spontaneous brain activity differed between TAO with and without DON, which may reflect the underlying pathological mechanism of DON. The ReHo index can be considered a diagnostic biomarker. CLINICAL RELEVANCE STATEMENT: Spontaneous brain activity in DON differed from that in TAO without DON, which may reflect the underlying pathological mechanism of DON. The ReHo index can be considered a diagnostic biomarker for early detection of DON. KEY POINTS: • Dysthyroid optic neuropathy (DON) affects brain activity, which contributes in the understanding of its visual dysfunction. • Regional homogeneity values differ between thyroid-associated ophthalmopathy with and without DON in various brain regions. • Regional homogeneity values can be used as a biomarker in the differential diagnosis of DON.

Utility of isolated-check visual evoked potential technique in dysthyroid optic neuropathy.

PURPOSE: To analyze the utility of isolated-check visual evoked potential (icVEP) for discriminating between eyes with dysthyroid optic neuropathy (DON) and eyes with thyroid-associated ophthalmopathy (TAO) but not DON. METHODS: Forty-three eyes with TAO but not DON (as non-DON), fifty-three eyes with DON, and sixty healthy eyes (as controls) were included. Comprehensive ophthalmic examinations, including best-corrected visual acuity, refraction, color vision test, intraocular pressure measurement, slit-lamp biomicroscopy, ophthalmoscopy, RAPD, exophthalmometry measurements, pVEP test, icVEP test, standard automated perimetry, and clinical activity score classification of TAO, as well as demographic information, were collected and analyzed. RESULTS: In the DON group, the signal-to-noise ratio (SNR) value of icVEPs decreased significantly compared with that of the non-DON group as well as control (p < 0.05). The SNR values under 8%, 16% and 32% depth of modulation (DOM) were significantly negatively correlated with BCVA (p < 0.05, r = - 0.9 ~ - 0.6), papilledema (Y/N) (p < 0.05, r = - 0.8 ~ 0.4) and DON (Y/N) (p < 0.001, r = - 0.7 ~ - 0.5). The 8% DOM of icVEP had the largest area under the receiver operating characteristic curve (AUC) (0.842) for discriminating DON from non-DONs. Meanwhile, decision curve analysis (DCA) showed that patients clinically benefit most from 8% DOM of icVEP. Furthermore, the 8% DOM of icVEP combing with papilledema (Y/N) and BCVA (Model 1) has significantly larger AUC than the 8% DOM of icVEP (p = 0.0364), and has better clinical benefit in DCA analysis. CONCLUSIONS: The SNR of 8% DOM from icVEP may represent a significant ancillary diagnostic method for DON detection. Furthermore, icVEP combined with papilledema (Y/N) and BCVA should be considered as a diagnostic model in future clinical practice.

Changes in the gut microbiota of patients with Graves' orbitopathy according to severity grade.

BACKGROUND: To explore the changes of gut microbiota in Graves' orbitopathy (GO) patients of different severity grades and to identify the pathogenic bacteria of GO and the associated mechanism. METHODS: A total of 18 healthy controls and 62 GO patients were recruited. The baseline information and faecal samples of all subjects were collected for gut microbiota analysis and metabolic function prediction analysis. 16SrDNA sequencing was used for microbial diversity detection. The operational taxonomic unit (OTU) was divided using the Mothur software, and the dominant microbiota was analysed. OTU number, Chao1 index, ACE index, and Shannon index of microbiota in faecal samples were analysed using the QIIME1.9.0 software. The relative abundance of microbiota in faecal samples was analysed through principal component analysis (PCA) using the Canoco Software 5.0. The metabolic function of microbiota in faecal samples was predicted using PICRUSt 2.0. RESULTS: There was no remarkable difference in gut microbiota diversity between groups; however, the gut microbial community and dominant microbiota significantly differed among groups. Klebsiella_pneumoniae was deemed the potentially pathogenic bacteria of GO, and its abundance was positively correlated with disease severity. The metabolic prediction results revealed that inorganic nutrition metabolism, fatty acid and lipid degradation, electron transfer, aromatic compound degradation, and alcohol degradation were notably different between groups with high and low abundance of Klebsiella_pneumoniae and among groups with different GO severity grades, thereby showing a positive correlation with GO clinical risks. CONCLUSIONS: Klebsiella_pneumoniae was a potential GO-related pathogen, which may regulate the metabolic pathways to affect GO progression.

Botox-A improve the thyroid-associated ophthalmopathy (TAO) orbital fibroblast activation through inhibiting the TGF-ß/Smad signaling.

The activation of orbital fibroblasts can result in fibrosis, finally contributing to thyroid-associated ophthalmopathy (TAO) progression. Although the effect of BTX-A on the treatment of TAO-related strabismus and upper eyelid retraction has long been recognized in clinical work, the underlying mechanism of BTX-A improving TAO-related strabismus and upper eyelid retraction has not been uncovered yet. In the present study, we successfully isolated and authenticated normal and TAO orbital fibroblasts. Compared with PBS, BTX-A and TACA exerted similar inhibitory effects on TAO orbital fibroblast proliferation and ECM production. TGF-ß stimulation induced the proliferation and ECM production by TAO orbital fibroblast, which was significantly inhibited by BTX-A or TACA treatment. Under TGF-ß stimulation, the inhibitory effects of BTX-A or TACA treatment on TAO orbital fibroblast proliferation and ECM production were reversed by TGF-ß/Smad signaling agonist SRI-011381. Collectively, BTX-A inhibited TGF-ß-induced TAO orbital fibroblast activation through inhibiting the TGF-ß/Smad signaling. Considering that TACA shows no satisfactory curative effects on symptoms closely related to the function of extraocular muscles, such as eye movement and diplopia, BTX-A might be a promising agent in TAO treatment.

Tauopathy induces degeneration and impairs regeneration of sensory nerves in the cornea.

The cornea is transparent and innervated by a dense collection of sensory nerves originating from the ocular branch of the trigeminal nerve. This study was designed to comprehensively analyze alterations of corneal sub-basal nerve plexus in a mouse model of tauopathy (P301L transgenic mice) to test the possibility of using corneal nerves as a biomarker for tauopathy. Corneal sensitivity, thickness and epithelial wound healing were measured non-invasively by aeshesiometer, optical coherence tomography and fluorescein staining, respectively. Tau, corneal nerves and immune cells were examined by immunohistochemistry or Western blot. At the early stage of tauopathy, although corneal sensitivity, thickness and nerve fiber density were not greatly altered, corneal nerve abnormalities were observed in the peripheral region of young P301L mice. With aging, the density of abnormal nerves increased, while corneal sensitivity, epithelial thickness, nerve fiber density and length decreased in middle-aged P301L mice compared with WT mice. After corneal epithelial injury in young mice, no difference in reepithelialization was observed between two groups of mice, however, the regeneration of corneal nerves in P301L mice lagged behind WT mice, which was reflected by delayed recovery of corneal sensitivity, decreased corneal nerve density and length and density of CD45+ dendriform cells in P301L mice. In conclusion, our data provide compelling evidence that corneal nerves were changed in a mouse model of tauopathy in an age-dependent manner. Moreover, tau overexpression impairs corneal nerve regeneration. These results suggest that cornea may serve as a promising ocular site for the early diagnosis of tauopathy.

CXCR3 deletion aggravates corneal neovascularization in a corneal alkali-burn model.

Corneal neovascularization can cause devastating consequences including vision impairment and even blindness. Corneal inflammation is a crucial factor for the induction of corneal neovascularization. Current anti-inflammatory approaches are of limited value with poor therapeutic effects. Therefore, there is an urgent need to develop new therapies that specifically modulate inflammatory pathways and inhibit neovascularization in the cornea. The interaction of chemokines and their receptors plays a key role in regulating leukocyte migration during inflammatory response. CXCR3 is essential for mediating the recruitment of activated T cells and microglia/macrophages, but the role of CXCR3 in the initiation and promotion of corneal neovascularization remains unclear. Here, we showed that the expression of CXCL10 and CXCR3 was significantly increased in the cornea after alkali burn. Compared with WT mice, CXCR3-/- mice exhibited significantly increased corneal hemangiogenesis and lymphangiogenesis after alkali burn. In addition, exaggerated leukocyte infiltration and leukostasis, and elevated expression of inflammatory cytokines and angiogenic factor were also found in the corneas of CXCR3-/- mice subjected to alkali burn. With bone marrow (BM) transplantation, we further demonstrated that the deletion of CXCR3 in BM-derived leukocytes plays a key role in the acceleration of alkali burn-induced corneal neovascularization. Taken together, our results suggest that upregulation of CXCR3 does not exhibit its conventional action as a proinflammatory cytokine but instead serves as a self-protective mechanism for the modulation of inflammation and maintenance of corneal avascularity after corneal alkali burn.

Clinical relevance of serum immunoglobulin G4 in glucocorticoid therapy of Graves' ophthalmopathy.

OBJECTIVE: Previous study suggested IgG4 levels were associated with the development of Graves' ophthalmopathy (GO). The aims of the present study were to investigate the role of IgG4 levels in glucocorticoid (GC) treatment in GO patients. DESIGN: 69 GO patients were enrolled. Serum thyroid hormones, thyroid antibodies, IgG, IgG4, ophthalmological examinations and orbital MRI were performed. Furthermore, the clinical outcomes (a composite response endpoint including the clinical activity score (CAS), proptosis, vision, intraocular pressure, diplopia and lid width) after high-dose intravenous GC treatment in 32 active moderate-to-severe GO patients were compared. PATIENTS: 69 consecutive patients with GO were asked to participate in the study. 32 of 69 GO patients were treated with high-dose intravenous GCs. MEASUREMENTS: Measurement of serum IgG and IgG4, serum thyroid hormones and thyroid autoantibodies. An overall ophthalmic assessment was performed pretherapy (week 0) and post-therapy (week 12). RESULTS: 33.3% of GO patients (23/69) had elevated IgG4 levels. IgG4 levels were positively correlated with the severity and activity of GO. After GC therapy, IgG4, IgG4/IgG, vision and CAS were significantly improved in GO patients. Patients with high IgG4 levels had a significantly reduced extraocular muscle area (EOMs) and better clinical outcomes than patients with normal IgG4 levels. CONCLUSIONS: Our results suggest a possible subgroup of elevated IgG4 GO patients, with more severe ophthalmopathy and better response to GCs treatment compare with normal IgG4 GO patients.

The diagnostic value of the IDEAL-T2WI sequence in dysthyroid optic neuropathy: a quantitative analysis of the optic nerve and cerebrospinal fluid in the optic nerve sheath.

OBJECTIVES: To evaluate the optic nerve and CSF in the optic nerve sheath as imaging markers of dysthyroid optic neuropathy (DON). METHODS: In this single-centre retrospective study, orbital images of 30 consecutive participants (54 orbits) with DON, 30 patients (60 orbits) with thyroid-associated ophthalmopathy (TAO) without DON, and 19 healthy controls (HCs; 38 orbits) were analysed. The diameter and cross-sectional area of the optic nerve and its sheath, water fraction of the optic nerve, and volume of the fluid in the optic nerve sheath were measured and compared. The associations between MR parameters and clinical measures were assessed using correlation analysis. RESULTS: The diameter and water fraction of the optic nerve (3 mm and 6 mm behind the eyeball), optic nerve subarachnoid space (ONSS) (3 mm and 6 mm behind the eyeball), and subarachnoid fluid volume in the optic nerve sheath were significantly greater in the DON group than in the TAO group (p < 0.01) or HC group (p < 0.01). ROC analysis showed that ONSS 3 mm behind the eyeball (ONSS3) was a robust predictor of DON (AUC = 0.957, sensitivity = 0.907, specificity = 0.9). Water fraction of the optic nerve 3 mm behind the eyeball (water fraction3) had the best specificity (0.967). Water fraction3, fluid volume in the optic nerve sheath, and optic nerve diameter (3 mm behind the eyeball) were correlated with clinical measures (i.e. clinical activity score, mean defect, and pattern standard deviation). CONCLUSIONS: Increased water fraction of the optic nerve and ONSS3 are promising and easily accessible radiological markers for diagnosing DON. KEY POINTS: • The water fraction of the optic nerve and optic nerve subarachnoid space (ONSS) are greater in patients with dysthyroid optic neuropathy (DON) than in patients with thyroid-associated ophthalmopathy (TAO) without DON. • The optic nerve and the cerebrospinal fluid in the optic nerve sheath measures are associated with visual dysfunction. • The water fraction of the optic nerve and ONSS may be promising imaging markers for diagnosing DON.

Imaging and anatomical parameters of the lacrimal punctum and vertical canaliculus using optical coherence tomography.

Purpose: The anatomical parameters of normal lacrimal puncta and vertical canaliculus using optical coherence tomography (OCT) and the OCT imaging features of punctal lesions were analyzed to provide a basis for clinical diagnosis and treatment. Methods: From June to September 2019, 40 volunteers (80 eyes) from Tongji Hospital were enrolled. The external punctal diameter (ELP) was measured using slit-lamp microscopy and OCT. The internal lacrimal punctal diameter (ILP) at 100 µm, vertical canalicular length (VCL), and tear meniscus depth were measured by OCT with open eyes. Twenty-eight volunteers (56 eyes) underwent the same examinations with their eyes closed. The OCT imaging features of 26 patients (27 eyes) with lacrimal lesions were examined. Results: The ELP of the right and left healthy eyes under slit-lamp microscopy were 564.40 and 555.40 µm respectively. Under OCT, the ELP, ILP, and VCL of the right and left eyes were 628.20 um and 616.85 µm, 343.40 µm and 346.95 µm, 731.95 um and 709.20 µm respectively. The ELP was larger when measured by OCT than slit-lamp microscopy (p<0.05). Twenty-eight volunteers (56 eyes) had measurements taken under different conditions. The ELP, ILP, and VCL of the open and closed right eyes were 667.54 and 567.21 µm, 369.18 and 303.18 µm, 715.00 and 417.14 µm, respectively. The ELP, ILP, and VCL of the open and closed left eyes were 655.86 um and 551.68 µm, 369.25 um and 313.54 µm, 719.96 um and 433.89 µm respectively. The anatomical parameters of the open eyes were greater than those of the closed eyes (p<0.05). Thus, we identified the imaging features of lacrimal stenosis, punctal obstruction, punctal tear, lacrimal atresia, and lacrimal mass using OCT. Conclusions: OCT can be used to measure the anatomical parameters of lacrimal puncta and vertical canaliculus in vivo. In addition, OCT can detect punctal lesions in vivo and provide an objective basis for the clinical diagnosis and treatment of punctal lesions.

Detection & analysis of inflammatory cytokines in tears of patients with lacrimal duct obstruction.

Background & objectives: Tear proteomic changes can be a candidate etiopathogenesis of lacrimal duct obstruction diseases (LDODs). Studies on proteomics have focused primarily on nasolacrimal duct obstruction, and some specific inflammatory cytokines such as interferon (IFN)-α2a, interleukin (IL)-8 and IL-10, have not been investigated. In addition, differences in inflammatory cytokines in tears according to the LDOD subtype have not been reported. This study aimed to quantitatively compare inflammatory cytokines in tears from patients with LDOD and investigate tear-cytokine differences among different LDOD subtypes. Methods: Tear samples were collected from both eyes of 30 patients with unilateral LDOD: five patients with prelacrimal obstruction, five with acute dacryocystitis and 20 with chronic dacryocystitis. The contralateral eyes were used as controls. IFN-α2a, IFN-ß, IFN-γ, IL-17A, IL-6, IL-8, tumour necrosis factor-alpha (TNF-α), vascular endothelial growth factor (VEGF)-A, induced protein-10 (IP-10) and monocyte chemotactic protein-1 (MCP-1) were quantified in all samples. Results: The expression of eight cytokines (except for IP-10 and MCP-1) were significantly increased in the affected eyes compared with those in the control eyes. The levels of nine inflammatory cytokines (except for IP-10) in the affected eyes of patients with chronic dacryocystitis were higher than those in the affected eyes of patients with prelacrimal obstruction. In addition, patients with chronic dacryocystitis presented significantly higher IFN-γ level than those with prelacrimal obstruction or acute dacryocystitis. Interpretation & conclusions: Specific pro-inflammatory cytokines were increased in tears of patients with LDOD compared with those in the controls. The specific cytokine profiles observed in the tears of individuals with different LDOD subtypes may be associated with the unique aetiopathogenesis of these conditions.

The microbiologic spectrum of dacryocystitis.

BACKGROUND: To investigate the microbiologic spectrum of dacryocystitis in adult and pediatric groups, specifically the microbiologic differences between chronic dacryocystitis with nasolacrimal duct obstruction (NLDO) and acute dacryocystitis in pediatric group. METHODS: This retrospective study was reviewed for demographic and microbiologic profile of dacryocystitis. The culture results were reported. RESULTS: Sixty-four adults and one hundred and five pediatrics with dacryocystitis were included in this study. Of all adults, only chronic dacryocystitis with NLDO was observed. Of all pediatric patients, 89 had chronic dacryocystitis with NLDO and 16 had acute dacryocystitis. Gram positive and negative isolates were numerically equal in adult group (both 36(48.65%)), while gram positive isolates were the major organism in pediatric group (71(58.68%)). Streptococcus pneumonia was the most common isolate in both adult (11(14.86%)) and pediatric (30(24.79%)) dacryocystitis. For both pediatric subgroups, gram positive isolates were the major organism (59(57.84%) for chronic dacryocystitis with NLDO and 12 (63.16%) for acute dacryocystitis). However, the leading isolates in those two subgroups were distinct, with Streptococcus pneumonia (29(28.43%)) being most common in chronic dacryocystitis with NLDO and Staphylococcus aureus (8(42.11%)) being most common in acute dacryocystitis. CONCLUSIONS: In adult group, gram negative isolates were more common in dacryocystitis than before. In pediatric group, gram positive isolates were still the major infection pathogen. Moreover, the more virulent organisms were more common in acute dacryocystitis than chronic dacryocystitis with NLDO.

Utility of 80-MHz Ultrasound Biomicroscopy and Lacrimal Endoscopy in Chronic Lacrimal Canaliculitis.

Lacrimal canaliculitis (LC) is a rare infection of lacrimal passage, which is usually late diagnosed or misdiagnosed. Traditional lacrimal system tests barely provide a clear and definite understanding of the pathological changes in lacrimal passage. We presented three patients with asymptomatic and atypical symptoms who were misdiagnosed and were eventually diagnosed with chronic LC with assistance of 80-MHz ultrasound biomicroscopy (80-MHz UBM) and lacrimal endoscopy. To our knowledge, the mutual assistance of above two techniques diagnosing LC has never been reported, it can provide better images and observations of the canaliculus from the inside out and can guide the differential diagnosis.

Characteristics of lacrimal passage diseases by 80-MHz ultrasound biomicroscopy: an observational study.

PURPOSE: To investigate the microstructure of the lacrimal canaliculus and the characteristics of lacrimal canalicular diseases by 80-MHz ultrasound biomicroscopy (UBM). METHODS: This study included 33 participants: 20 normal subjects (40 eyes), 2 patients with chronic lacrimal canaliculitis (4 eyes), 10 patients with chronic dacryocystitis (16 eyes), and 1 patient with lacrimal punctum atresia (2 eyes). All participants underwent 80-MHz UBM; disease-specific features were noted. RESULTS: On 80-MHz UBM of the lacrimal canaliculi (vertical section) in normal subjects, low echo of the lacrimal canalicular lumen and high echo of the lacrimal canalicular wall were observed. The uniform low echo near the wall was the mucosal epithelium. The outermost layer of medium-to-high echo was the subepithelial elastic fibrous layer. In the horizontal section, the lumen was continuous. Two linear high echoes parallel to the canalicular wall could be observed at the center of the lacrimal canaliculus, which were sometimes attached and sometimes separated. When separated, the center of the lacrimal canaliculus was a low echo area (lumen). Lacrimal canaliculitis (vertical section) showed obvious ectasia of the lacrimal canalicular lumen, with a high echo mass shadow, which might have been calculi, and uneven thickness of the mucosal epithelium with a slightly high echo shadow. In the horizontal section, the lumen varied in size with clear boundaries of medium and high echoes. The central linear high echoes of the lumen were absent, and the echoes of the mucosal epithelium were discontinuous. In chronic dacryocystitis, the lacrimal canalicular lumen was extensively enlarged, with continuous echoes and uniform thickness of the mucosal epithelium and homogeneous patches of slightly higher echoes. Lacrimal punctum atresia indicated that the lacrimal canaliculus existed in both eyes and its structure was normal. CONCLUSIONS: The 80-MHz UBM is a new non-invasive technique that can be used for clear visualization of the fine structure of the lacrimal canaliculus, including the mucosal epithelium and subepithelial elastic fiber layer. The use of this approach will improve understanding of the hierarchical structure of the lacrimal canaliculi and provide a comprehensive basis for diagnosis, differential diagnosis, and treatment plan in patients with lacrimal passage diseases.

Pathological changes of the nasolacrimal duct in rabbit models of chronic dacryocystitis: correlation with lacrimal endoscopic findings.

PURPOSE: The aim of this study was to explore the pathological changes of the nasolacrimal duct in rabbits with experimentally induced obstructive dacryocystitis in correlation with lacrimal endoscopic findings. METHODS: The rabbit model of obstructive dacryocystitis was created by injecting 0.15 ml of self-curing resin into the lacrimal duct. The control group received 0.15 ml of normal saline. Within 16 weeks after the obstructive, lacrimal endoscopy and pathological examination of the nasolacrimal duct were conducted at different time points of 1, 2, 4, 8, and 16 weeks. RESULTS: In the control group, lacrimal endoscopy revealed pink and smooth mucosa; and the pathological analysis revealed an epithelial layer that was composed of superficial columnar cells and a deep basal epithelial layer. The experimental rabbits showed clinical manifestations of obstructive dacryocystitis a week after the injection of self-curing resin. At weeks 1 and 2, the lacrimal endoscopy showed mucosal hyperemia and hemorrhagic spots on the nasolacrimal duct; and the pathological features included epithelial cell swelling and inflammatory cell infiltration. At weeks 4 and 8, the experimental group showed alternatively red and white mucosa under the lacrimal endoscopy, and the pathological features included proliferative epithelium accompanied by papillary hyperplasia. At week 16, the experimental group showed pale and coarse mucosa and white membrane-like layer covering the mucosal surface, and the pathological features included epithelial necrosis, squamous metaplasia, and sub-epithelial fibrosis. CONCLUSION: The mucosa of the nasolacrimal duct showed different pathological features at different time points after lacrimal duct obstruction, which was well correlated with the endoscopic findings. It is possible to predict the pathological stages by the endoscopic observation in NLOD patients.

Genetically engineered probiotics for an optical imaging-guided tumor photothermal therapy/immunotherapy.

Bacteria-based cancer therapy (BCT) has been extensively investigated because of the tumor targeting and antitumor immunity activating abilities of bacteria over traditional nanodrugs, but their potential systemic toxicity poses a challenge. Therefore, it is important to visualize the precise localization and real-time distribution of bacteria in vivo to guide the treatment. Herein, biogenetically engineered Escherichia coli Nissle 1917 (EcN) were constructed to highly express tyrosinase to intracellularly generate cyanine 5-labeled melanin-like polymers (Cy5-Mel), thus endowing them with a bright fluorescence and an excellent photothermal performance upon NIR laser irradiation, thereby inducing the intense immunogenic death of tumor cells and release of tumor-associated antigens. Acting as adjuvants, bacteria can greatly stimulate the maturation of dendritic (DC) cells. The in vivo behaviors of these bacteria was monitored via noninvasive optical imaging when they were intravenously administrated to tumor-bearing mice. From this, NIR exposure on tumor sites was carried out at an appropriate time point to induce the damage to tumor cells and for the modulation of tumor immune microenvironments. Thus, via a simple bioengineering strategy, a promising bacteria-based theranostic platform was constructed for tumor treatment.