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.

Correlation Analysis between Intraocular Pressure and Extraocular Muscles Based on Orbital Magnetic Resonance T2 Mapping in Thyroid-Associated Ophthalmopathy Patients.

BACKGROUND: The correlation between intraocular pressure (IOP) and the magnetic resonance imaging (MRI) parameters in thyroid-associated ophthalmopathy (TAO) patients was explored. METHODS: This study included 82 eyes in 41 TAO patients who had a large difference in the IOP between each eye. We measured the T2 relaxation time (T2RT) of the extraocular muscles (EOMs), the orbital fat, and the area of the EOMs. RESULTS: There was a positive correlation between IOP and exophthalmos, the clinical activity score (CAS), the T2RT (of the medial rectus (MR)), the area of the MR, inferior rectus (IR) and lateral rectus, and the mean area. We established a regression model with IOP as the dependent variable, and the area of the IR was statistically significant. CONCLUSIONS: High IOP in TAO patients was positively correlated with the degree of exophthalmos and EOM inflammation (especially the inferior rectus). The state of the EOMs in an orbital MRI may partially explain high IOP and provide the necessary clinical information for subsequent high IOP treatment.

Ocular Manifestations and Clinical Implications of Serum Immunoglobulin G4 Levels in Graves' Ophthalmopathy Patients.

PURPOSE: To investigate the association between serum immunoglobulin G4 (IgG4) levels and laboratorial parameters. Moreover, the ocular manifestations were compared in Graves' ophthalmopathy (GO) patients. METHODS: Fifty-seven GO patients and fifty sex- and age-matched euthyroid subjects were enrolled. Serum IgG4 and immunoglobulin G (IgG) levels, thyroid hormones and thyroid autoantibodies were measured in all participants. Ophthalmologic examinations were performed to GO patients. RESULTS: Serum IgG4 levels were increased in GO patients compared with euthyroid subjects (p<0.001). Multivariate analysis showed that TRAb levels were correlated with IgG4 levels in GO patients (ß=1.45, p = 0.024). There were eight GO patients with IgG4 levels ≥ 135 mg/dL. They all had both eyelids swelling and the differences of HER were less than 2mm. Seven patients (7/8) showed bilateral symmetrical thickening of the extraocular muscles. CONCLUSIONS: Bilateral symmetry of ocular manifestation was observed in GO patients with IgG4 levels ≥135 mg/dL. IgG4 levels may be helpful in monitoring GO and diagnosing a new subtype of GO.

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.

Correlations Between Single-Photon Emission Computed Tomography Parameters of Salivary Glands and Dry Eye Tests in Patients With Sjögren's Syndrome.

OBJECTIVE: To evaluate the correlations between Single-Photon Emission Computed Tomography (SPECT) parameters of salivary glands and dry eye parameters in patients with Sjögren's syndrome (SS). METHODS: A total of 28 patients with SS participated in this prospective study. Dry eye assessments include tear film break-up time (TBUT), corneal fluorescein staining scoring (CFS), Schirmer's I test (SIT) examination and SPECT of salivary gland. The following quantitative parameters were derived from SPECT imaging for salivary glands: Uptake index (UI), the time needed to achieve the minimum counts after Vit C stimulation (Ts), and excretion fraction (EF). The relation between the aforementioned parameters and TBUT, CFS and SIT were analyzed with SPSS 22.0 software. RESULTS: All the 28 eyes of the 28 subjects were examined. The mean SIT was 6.04 ± 4.64 mm/5 min (0-18 mm/5 min); the mean CFS was 3.07 ± 2.65 (0-10) and the mean BUT was 2.11 ± 1.97 s (0-9 s). The mean EF value was 0.52 ± 0.12 (0.26-0.75) in parotid glands and 0.45 ± 0.10 (0.30-0.67) in submandibular glands, respectively. The mean UI value was 9.33 ± 1.68 (6.03-13.20) in parotid glands and 9.92 ± 1.48 (7.08-12.60) in submandibular glands, respectively. The mean Ts (min) was 5.32 ± 3.01 (2.00-12.00) in parotid glands and 11.09 ± 7.40 (2.00- 29.00 min) in submandibular glands, respectively. It was found that EF positively correlates with SIT in patients with SS (r = 0.499 and 0.426 in parotid glands and submandibular glands, with P < 0.05), while no significant correlation was found between the UI, Ts and CFS, TBUT (P > 0.05). CONCLUSIONS: The EF was positively correlated with SIT in patients with SS, it could reflex the dysfunction of salivary glands in SS patients. So, EF may be a valuable parameter for the diagnosis of SS patients with lacrimal gland secretion dysfunction.

Comparing isolated-check visual evoked potential, pattern visual evoked potential, and standard automated perimetry in dysthyroid optic neuropathy eyes.

OBJECTIVE: To determine the diagnostic ability of isolated-check visual evoked potential (icVEP), pattern visual evoked potential (pVEP), and standard automated perimetry (SAP) between dysthyroid optic neuropathy (DON) and thyroid-associated ophthalmopathy (TAO) without DON (non-DON). METHODS: This is a case-control study, 49 bilateral patients (26 DON and 23 non-DON) were included. icVEP, pVEP, and SAP were conducted in all the subjects, icVEP parameters compared were signal-to-noise ratios (SNRs) under 8, 16, and 32% depth of modulation (DOM). pVEP parameters compared were amplitude and latency. SAP parameters were mean deviation (MD) and pattern standard deviation (PSD). The area under the receiver operating characteristic (ROC) curve (AUC), net reclassification index (NRI), integrated discrimination index (IDI), and decision curve analysis (DCA) were applied for analysis. RESULTS: In icVEP, values of SNR in DON were significantly smaller than non-DON (p < 0.05). In pVEP, P100 latent time in DON was significantly larger than non-DON (p = 0.0026). In SAP, value of PSD in DON was larger than non-DON (p = 0.0006), and value of MD in DON was smaller (p = 0.0007). AUC, NRI, and IDI among the three tests were not significantly different. DCA showed that SNR of icVEP under 8% DOM was the farthest from the two extreme curves. CONCLUSIONS: icVEP, pVEP, and SAP have equal diagnostic capabilities to discern between DON and non-DON. In addition, icVEP may represent a significant ancillary diagnostic approach to DON detection, with more clinical benefit.

Multiple cavernous hemangiomas in the orbit: A case report and review of the literature.

RATIONAL: Cavernous hemangiomas are one of the most common benign primary orbital lesions. These tumors are insidious in onset, slowly progressive and present more often in middle aged women. Multiple orbital cavernous hemangiomas are extremely rare, and only a few cases have been reported in the published literature. PATIENT CONCERNS: Here, we report the diagnosis and treatment of multiple cavernous hemangiomas in the right orbit of a female patient with impaired visual acuity and proptosis of the eye for more than 10 years. DIAGNOSIS: Magnetic resonance imaging of the orbit showed a giant and irregular soft mass filling the intraconal and extraconal space of the right orbit, compressing the right optic nerve. After tumor resection, histopathological examination confirmed the diagnosis of cavernous hemangioma. INTERVENTIONS: A lateral orbitotomy was performed and a total of 13 tumors were excised, with the largest tumor measuring approximately 2.5 × 3.0 cm. OUTCOMES: The visual acuity of the patient was preserved, with only a slightly dilated pupil of the right eye. The follow-up period was 6 months with no signs of recurrence. LESSONS: Multiple cavernous hemangiomas in the orbit is rare and should be excised surgically as soon as possible.

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.

A New Isolation Method of Human Lacrimal Canaliculus Epithelial Stem Cells by Maintaining Close Association with Their Niche Cells.

Purpose: To investigate whether lacrimal canaliculus epithelial stem cells (LCESC) could be isolated and expanded in vitro. Methods: The lacrimal canaliculus epithelium of 6 patients with limbal stem cell deficiency (LSCD) caused by alkali burn or Stevens Johnson Syndrome were examined by lacrimal endoscope. Cadaveric eyelids were fixed and prepared for cross section and stained with HE and antibodies against PCK, Vim, p63α, SCF and c-Kit. Canaliculus tissue was separated under an operating microscope using a lacrimal probe as an indicator and digested with collagenase A. The clusters of epithelial cells with closely associated stroma were further digested with Trypsin/EDTA to obtain single cells for culture on Matrigel-coated plastic plates in MESCM media. The expression of SCF, c-Kit and p63α was determined by immunostaining. The colony-forming efficiency on 3T3 feeder layers was also measured by calculating the percentage of the clone number divided by the total number cells seeded. Results: The epithelial layers of five out of six inferior lacrimal canaliculi and all the six superior lacrimal canaliculi were visually normal in appearance. Five to fifteen layers of the epithelium in the human lacrimal canaliculi were present with a small, tightly compacted basal layer of cells expressing PCK, p63α, SCF and c-Kit. LCESC were isolated by collagenase A and obtained clonal growth in MESCM. The colony-forming efficiency of LCESC holoclones on a 3T3 feeder layer was 3.2%, compared to 1.9% for those of limbal stem cells (LSC). Conclusions: Herein, we first report that LCESCs can be isolated and have stem cell characteristics, similar to those of LSCs. Such a discovery raises a promising substrate resource of stem cells for LSC reconstruction in LSCD patients.

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.

Three dimensional orbital magnetic resonance T2-mapping in the evaluation of patients with Graves' ophthalmopathy.

The clinical application of orbital magnetic resonance (MR) T2-mapping imaging in detecting the disease activity of Graves' ophthalmopathy (GO), and the predictive values of therapy response to intravenous glucocorticoid (ivGC) were investigated. Approved by the local institutional review board (IRB), 106 consecutive patients with GO were included in this prospective study. All subjects were divided into two groups according to the patients' clinical activity score (CAS): the CAS positive group (CAS ≥3) or the CAS negative group (CAS <3). T2 relaxation time of extraocular muscles (T2RT; ms) and the areas of four extra-ocular muscles (AEOMs; mm2) were measured by 3D T2-mapping MR sequence before and after methylprednisolone treatment, so as the CAS and some ophthalmic examinations including visual acuity, intra-ocular pressure, eyeball movement, diplopia and proptosis. In addition, 24 healthy volunteers were recruited as the control group. The mean T2RT and AEOMs in CAS positive group were higher than those in CAS negative group. Both CAS positive and negative groups had significantly higher mean T2RT and AEOMs than the control group (P<0.01). There was a positive correlation between T2RT and AEOMs values in GO patients, both of them had a positive correlation with CAS and the ophthalmic examinations. It was concluded that to evaluate the activity of GO, CAS was mostly related to inflammation symptoms of ocular surface, more than that, T2RT and AEOMs were also related to abnormal findings of the ophthalmic examinations including high ocular pressure, impaired eyeball movement, diplopia and proptosis. T2RT and AEOMs can reflex the inflammation state of ocular muscles better. CAS combined with 3D T2-mapping MR imaging could improve the sensitivity of detection of active GO so as the prediction and evaluation of the response to methylprednisolone treatment.

Modeling Chronic Dacryocystitis in Rabbits by Nasolacrimal Duct Obstruction with Self-Curing Resin.

We established a chronic dacryocystitis model by injecting of 0.05, 0.1, and 0.15 ml self-curing resin via the lacrimal punctum in rabbits. Animals were randomized into four groups (n = 11 animals/group). The control group received 0.15 ml normal saline. Within three months postinjection, epiphora and eye discharge were observed. At the 90th day postlacrimal passage irrigation, CT dacryocystography was performed to find changes in the lacrimal image, and hematoxylin and eosin staining was made to identify pathological changes of the lacrimal sac. Three months postinjection, the rabbits in control group and those who received 0.05 and 0.1 ml self-curing resin failed to develop chronic dacryocystitis. However, 8/11 (72.7%) rabbits those received 0.15 ml self-curing resin were symptomatic and showed complete reflux in lacrimal passage irrigation, indicating the obstruction of the nasolacrimal duct. CT dacryocystography showed that the obstruction was present only in the animals with chronic dacryocystitis. Pathological examinations of chronic dacryocystitis also revealed significantly inflammatory changes, such as mucus epithelium thickening, irregular papillary proliferation, and submucosal fibrous deposition. Local injection of 0.15 ml self-curing resin can induce permanent obstruction of the nasolacrimal duct in rabbits and establish a model of chronic dacryocystitis.

Intervention using a novel biodegradable hollow stent containing polylactic acid-polyprolactone-polyethylene glycol complexes against lacrimal duct obstruction disease.

Lacrimal duct obstruction disease (LDOD) is a common ophthalmologic disease. Stent implantation surgery is one of the most effective therapies. In this study, we intended to find out the satisfactory biodegradable stents containing poly-L-lactic acid-polycaprolactone-polyethylene glycol (PLLA- PCL- PEG) complexes for therapeutic application in LDOD. Stents made of PLLA- PCL- PEG complexes in various ratios, were prepared and used in vitro to determine stents with appropriate mechanical properties and shorter range of bio-degradation for study in vivo. Thirty-two rabbits were randomized into eight groups of four eyes each in advance for test in vivo. The selected stents were implanted into the left lacrimal ducts of 16 rabbits and silica gel stents as the control for the other 16 rabbits. At four points in time (1, 4, 10 and 16 weeks after the implantation), weight loss rate (WLR) of the stents was measured and analysed. To access the change of lacrimal duct, fluorescein excretion test, lacrimal duct endoscopy and histopathological testing were conducted. The stent containing PLLA: PCL6: 4+ 15%PEG was selected for study in vivo. Analysis of weight loss rate (WLR), fluorescein excretion test, lacrimal duct endoscopy and histopathological testing indicated that the selected stent was biodegradable and caused minimal stimulation and earlier tissue restoration in the lacrimal epithelium compared with the silica gel stent used as the control. The study results suggest that the PLLA: PCL6: 4+ 15% PEG stent is a satisfactory biodegradable stent as a promising alternative for therapeutic application in LDOD, which showed tissue compatibility, biodegradation and adequate mechanical intensity.

Lacrimal Duct Occlusion Is Associated with Infectious Keratitis.

Background: To explore the prevalence of lacrimal duct obstruction in patients with infectious keratitis, and the necessity of lacrimal duct dredge in the treatment of human infectious keratitis. Methodology/Principle Findings: The design is prospective, non-control case series. Thirty-one eyes from twenty-eight continuous patients with infectious keratitis were included in this study. The presence/absence of lacrimal duct obstruction was determined by the lacrimal duct irrigation test. The diagnosis of infectious keratitis was made based on clinical manifestations, cornea scraping microscopic examination and bacterial/fungus culture. Diagnosis of viral keratitis was set up based on the recurrent history, deep neovascularization and typical outlook of the cornea scar. The treatment of keratitis included drugs, eye drops or surgery, while treatment of chronic dacryocystitis was lacrimal duct dredging with supporting tube implantation surgery. In the thirty-one eyes with infectious keratitis, fifteen suffered from fungal keratitis (48%), two bacterial keratitis (6%), and fourteen viral keratitis (45%). Eleven eyes (35%) from ten patients with infectious keratitis also suffered from lacrimal duct obstruction. In those cases, six eyes also suffered from lower canalicular obstruction, three nasolacrimal duct obstruction and chronic dacryocystitis, one a combination of upper and lower canalicular obstruction, one upper canalicular obstruction. After local and systemic applications of anti-bacterial, anti-viral, anti-fungal and anti-inflammatory drugs, twenty-eight eyes (90%) recovered within three weeks, while the ulceration of three patients required the lacrimal duct dredging and supporting tube implantation surgery for the healing. Conclusions: Herein, we first report that the prevalence of infectious keratitis is closely correlated to the occurrence of lacrimal duct obstruction. When both confirmed, simultaneous treatment of keratitis and lacrimal duct obstruction promptly is required. Further evaluation of mechanism, prevention and control of the diseases are warranted.

Efficacy and Safety of rAAV2-ND4 Treatment for Leber's Hereditary Optic Neuropathy.

Leber's hereditary optic neuropathy (LHON) is a mitochondrially inherited disease leading to blindness. A mitochondrial DNA point mutation at the 11778 nucleotide site of the NADH dehydrogenase subunit 4 (ND4) gene is the most common cause. The aim of this study was to evaluate the efficacy and safety of a recombinant adeno-associated virus 2 (AAV2) carrying ND4 (rAAV2-ND4) in LHON patients carrying the G11778A mutation. Nine patients were administered rAAV2-ND4 by intravitreal injection to one eye and then followed for 9 months. Ophthalmologic examinations of visual acuity, visual field, and optical coherence tomography were performed. Physical examinations included routine blood and urine. The visual acuity of the injected eyes of six patients improved by at least 0.3 log MAR after 9 months of follow-up. In these six patients, the visual field was enlarged but the retinal nerve fibre layer remained relatively stable. No other outcome measure was significantly changed. None of the nine patients had local or systemic adverse events related to the vector during the 9-month follow-up period. These findings support the feasible use of gene therapy for LHON.

Galectin-9 ameliorates respiratory syncytial virus-induced pulmonary immunopathology through regulating the balance between Th17 and regulatory T cells.

Respiratory syncytial virus (RSV) infections are characterized by lung inflammation, mucus hypersecretion, and hyperresponsiveness. CD4+ T cells play a pivotal role in the development of RSV-induced lung pathology. Thus targeting the activation of CD4+ T cell subsets and enhancing regulatory functions of CD4+ T cells could be an effectively therapeutic approach. In the present study, we showed that RSV-induced lung inflammation can be suppressed by lectin family member Galectin-9 (Gal-9), which is identified as a T-cell immunoglobulin- and mucindomain-containing molecule-3 (Tim-3) ligand (L) and the Gal-9/Tim-3 interaction acts as a specific inhibitor of T helper(Th)1 and Th17 immune responses. Tim-3 expression was up-regulated in RSV-infected mice compared to non-infected controls. Therefore, we constructed a recombinant adenoviral (rAAV) 9-Gal-9 adenoviral plasmid, and administered it intranasally into RSV-infected mice for five times at every other day until day 8 post-infection. We found that Gal-9 administration significantly decreased viral load, inhibited mucus production, and diminished severity of lung pathology which were all induced by RSV infection. Complicated mechanisms were involved in these inhibitory effects, including inhibition of Th17 cell production, induction of regulatory cell expansion, as well as alteration of CD8 T-cell apoptosis. Our findings suggest that regulating the function of the Gal-9/Tim-3 pathway will be an effective and safe approach to treat RSV infection in lungs.

Promotion of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on titanium.

AIM: To investigate the influence of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on titanium (Ti) surface. METHODS: The chimeric peptide RKLPDAPRGDN (minTBP-1-PRGDN) was synthesized by connecting RKLPDA (minTBP-1) to the N-terminal of PRGDN, the influence of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on Ti surface were tested using PRGDN and minTBP-1as controls. The keratocytes attached to the surface of Ti were either stained with FITC-labeled phalloidin and viewed with fluorescence microscope or quantified with alamar Blue method. The proliferation of keratocytes on Ti were quantified with 3-(4,5-dim- ethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide up-taking methods. The secretion of type I collagen were determined using an ELISA kit. RESULTS: The results showed that minTBP-1-PRGDN at a concentration of 100ng/mL was the most potent peptide to enhance the attachment of human keratocytes to the surface of Ti (1.40±0.03 folds, P=0.003), to promote the proliferation (1.26±0.05 folds, P=0.014) and the synthesis of type I collagen (1.530±0.128, P=0.008). MinTBP-1 at the same concentration could only promote the attachment (1.13±0.04 folds, P=0.020) and proliferation(1.15±0.06 folds, P=0.021), while PRGDN had no significant influence (P>0.05). CONCLUSION: Our data shows that the novel chimeric peptide minTBP-1-PRGDN could promote the attachment, proliferation and type I collagen synthesis of human keratocytes on the surface of Ti.

Dexamethasone reduces IL-17 and Tim-3 expression in BALF of asthmatic mice.

This study investigated the expression of interleukin-17 (IL-17) and T cell immunoglobulin mucin and domain-containing molecule-3 (Tim-3) in bronchoalveolar lavage fluid (BALF) of asthmatic mice and the effect of dexamethasone (DEX) on these factors. Thirty-six mice were randomly divided into three groups: normal group, asthmatic group and DEX group. The mouse model of asthma was established by sensitization with ovalbumin in both the asthmatic and DEX groups. The levels of IL-6, IL-10, IL-17 and TGF-ß were measured in BALF by enzyme-linked immunesorbent assay (ELISA). The mRNA expression level of Tim-3 was detected by reverse transcription polymerase chain reaction (RT-PCR). The ratio of Tim-3+CD4+ cells to total CD4+ cells in BALF was determined by flow cytometry. Differential inflammatory cells in BALF were detected. The correlations among IL-17, IL-6, IL-10, Tim-3 and inflammatory cells were analyzed. The results showed that the levels of IL-17, IL-6 and Tim-3 were substantially increased and the IL-10 level decreased in BALF in the asthmatic mice, which was significantly reversed by DEX treatment. IL-17 expression was positively correlated with IL-6 and Tim-3 expression and the number of inflammatory cells but negatively with IL-10 expression. These results indicate that the increased expression of IL-17 and Tim-3 in BALF may be implicated in the occurrence and development of asthmatic inflammation; the mechanism by which DEX suppresses asthmatic airway inflammation involves down-regulation of IL-17 and Tim-3 levels.

Small interfering RNA targeting T-cell Ig mucin-3 decreases allergic airway inflammation and hyperresponsiveness.

Since CD4+ T cells play a pivotal role in the development of airway inflammation and hyperresponsiveness, targeting activated CD4+ T cell subsets and increasing the cells with regulatory function would be a logical therapeutic approach. We showed that this outcome can be achieved by local therapy with Tim-3, which is a negative regulator of CD4+ T cells. Tim-3 expression was up-regulated by ovalbumin (OVA) induction. Attenuating Tim-3 expression by RNA interference suppressed allergen-induced immune responses. Intranasal application of Tim-3 shRNA diminished airway inflammation and hyperresponsiveness. Multiple mechanisms were involved in the inhibitory effects, including regulation the imbalance of Th1/Th17 and increasing Treg cell expression. Our results indicate that the Tim-3 pathway is highly involved in the regulation of asthma. Targeting Tim-3 by siRNA may hold therapeutic potential in preventing the development of allergic asthma.