Tea Polyphenols Reduce Inflammation of Orbital Fibroblasts in Graves’Ophthalmopathy via the NF-κB/NLRP3 Pathway

Objective:This study aimed to explore the effects of tea polyphenols(TP)on inflammation of orbital fibroblasts in Graves’ophthalmopathy(GO)and to provide new ideas for GO treatment.Methods:Primary orbital fibroblasts were extracted from orbital adipose/connective tissues of patients with and without GO.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of interleukin(IL)-6,IL-1β,and monocyte chemotactic protein(MCP)-1 in non-GO and GO orbital fibroblasts.The CCK-8 assay was used to determine the appropriate concentration of TP for subsequent experiments.RT-qPCR and enzyme-linked immunosorbent assay(ELISA)were performed to investigate the effects of TP on lipopolysaccharide(LPS)-induced production of inflammatory cytokines.Nuclear factor-κB(NF-κB)expression was measured using Western blotting analysis.NOD-like receptor 3(NLRP3)expression was detected using both Western blotting analysis and immunofluorescence staining.Results:The mRNA levels of IL-6,IL-1β,and MCP-1 in GO orbital fibroblasts were significantly higher than those in non-GO cells.TP treatment significantly inhibited LPS-induced production of inflammatory factors,including IL-6,IL-1β,and MCP-1.TP also inhibited the expression levels of NF-κB and NLRP3.Inflammation in the GO orbital fibroblasts was higher than that in non-GO cells.TP inhibited the production of inflammatory cytokines in GO orbital fibroblasts in vitro through the NF-κB/NLRP3 pathway.Conclusion:These findings suggest that TP may have a potential role in GO treatment.

Association of Graves’ disease and Graves’ ophthalmopathy with the polymorphisms in promoter and exon 1 of cytotoxic T lymphocyte associated antigen-4 gene

Objective: To investigate the association of Graves’ disease and Graves’ ophthalmopathy with the C/T transition polymorphism at position –318 of promoter and the A/G transition polymorphism at position 49 of exon 1 within cytotoxic T lymphocyte associated antigen-4 (CTLA-4) gene. Methods: Thirty-three patients with ophthalmopathy of Graves’ disease, fifty-six Graves’ patients without ophthalmopathy and sixty normal subjects as control were involved in the present case-control study. The polymorphisms were evaluated by polymerase chain reaction fragment length polymorphism (PCR-RFLP). Com-parisons were made of gene frequencies and allele frequencies between the groups. Results: The gene frequencies of CT and allele frequencies of T were much higher in Graves’ patients with ophthalmopathy than that in the group without ophthalmopathy (P=0.020, P=0.019). The gene frequencies of GG and allele frequencies of G in patients with Graves’ disease were significantly increased as compared with control group (P=0.008, P=0.007). The data suggest that smokers with Graves’ disease seemed to be more predisposed to ophthalmopathy than non-smokers (P=0.018). Conclusion: Our results suggest that an allele of T at position –318 of promoter is associated with genetic susceptibility to Graves’ ophthalmopathy while an allele of G at position 49 of exon 1 is associated with genetic susceptibility to Graves’ disease instead. Smoking is believed to be a major risk factor for ophthalmo-pathy.

Orbital decompression surgery for the treatment of Graves＇ ophthalmopathy：comparison of different techniques and long-term results

AIM： To evaluate the long-term results of different orbital decompression techniques performed in patients with Graves＇ ophthalmopathy（GO）.METHODS： Totally 170 cases with GO underwent orbital decompression between 1994 and 2014. Patients were divided into 4 groups as medial-inferior, mediallateral（balanced）, medial-lateral-inferior, and lateral only according to the applied surgical technique. Surgical indications, regression degrees on Hertel exophthalmometer, new-onset diplopia in the primary gaze and new-onset gaze-evoked diplopia after surgery and visual acuity in cases with dysthyroid optic neuropathy（DON） were compared between different surgical techniques.RESULTS： The study included 248 eyes of 149 patients. The mean age for surgery was 42.3±13.2 y. DON was the surgical indication in 36.6% of cases, and three-wall decompression was the most preferred technique in these cases. All types of surgery significantly decrease the Hertel values（P〈0.005）. Balanced medial-lateral, and only lateral wall decompression caused the lowest rate of postoperative new-onset diplopia in primary gaze. The improvement of visual acuity in patients with DON did not significantly differ between the groups（P=0.181）. CONCLUSION： The study show that orbital decompression surgery has safe and effective long term results for functional and cosmetic rehabilitation of GO. It significantly reduces Hertel measurements in disfiguring proptosis and improves visual functions especially in DON cases.

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（iv GC） 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.

Ocular surface changes in Graves’ophthalmopathy

Many patients with Graves’ophthalmopathy(GO)suffer from dry eye syndrome(DES),and this is one of the most common reasons of eye discomfort in patients with GO.The prevalence of DES in patients with GO is significantly higher than normal subjects.The ocular surface changes involving changes in tears,cornea,conjunctiva and glands occur in GO patients.However,the mechanism of how DES occurs in GO still remains unclear.In this review,the ocular surface changes were illustrated and analyzed the reasons for high prevalence of DES in GO patients.

Association of Polymorphisms of rs179247 and rs12101255 in Thyroid Stimulating Hormone Receptor Intron 1 with an Increased Risk of Graves' Disease:A Meta-analysis

The polymorphisms of thyroid stimulating hormone receptor（TSHR） intron 1 rs179247 and rs12101255 have been found to be associated with Graves＇ disease（GD） in genetic studies. In the present study, we conducted a meta-analysis to examine this association. Two reviewers systematically searched eligible studies in Pub Med, Web of Science, Embase and China Biomedical Literature Database（CBM）. A meta-analysis on the association between GD and TSHR intron 1 rs179247 or rs12101255 was performed. The odd ratios（OR） were estimated with 95% confidence interval（CI）. Meta package in R was used for the analyses. Seven articles（13 studies） published between 2009 and 2014, involving 5754 GD patients and 5768 controls, were analyzed. The polymorphism of rs179247 was found to be associated with an increased GD risk in the allele analysis（A vs. G： OR=1.40, 95% CI=1.33–1.48） and all genetic models（AA vs. GG： OR=1.94, 95% CI=1.73–2.19; AA＋AG vs. GG： OR=1.57, 95% CI=1.41–1.74; AA vs. AG＋GG： OR=1.54, 95% CI=1.43–1.66）. The site rs12101255 also conferred a risk of GD in the allele analysis（T vs. C： OR=1.50, 95% CI=1.40–1.60） and all genetic models（TT vs. CC： OR=2.22, 95% CI=1.92–2.57; TT＋TC vs. CC： OR=1.66, 95% CI=1.50–1.83; TT vs. TC＋CC： OR=1.74, 95% CI=1.53–1.98）. Analysis of the relationship between rs179247 and Graves＇ ophthalmopathy（GO） showed no statistically significant correlation（A vs. G： OR=1.02, 95% CI=0.97–1.07）. Publication bias was not significant. In conclusion, GD is associated with polymorphisms of TSHR intron 1 rs179247 and rs12101255. There is no association between rs179247 SNPs and GO.

Analysis of Graves＇ ophthalmopathy patients＇ tear protein spectrum

Background Graves＇ ophthalmopathy/orbitopathy （GO） patients often suffer ocular surface damages and tear fluid proteins play a significant role in maintaining healthy ocular surfaces,while changes in tear protein components reflect the changes ocular surface abnormalities.In this study proteomics techniques were used to investigate tear protein compositions in GO patients.Methods We carried out a case-control study by comparing tear fluid contents of GO patients with that of healthy subjects.In the first step the tears were subjected to SDS-PAGE electrophoresis and then single protein bands were analyzed by to in-gel trypsin digestion and nano-flow liquid mass spectrometry （LC-MS/MS） using a MS software.Results In tear samples of GO subjects,the protein fractions of inflammation-related protein immunoglobulin kappa chain C region （IgKC） and serum albumin were essentially reduced,whereas a novel isoform of complement component 3 （C3）,which we detected in control subjects,was completely absent in the GO patients＇ tears.Conclusions Reduced protein concentrations of particularly IgKC and complement C3 as well as albumin in the tears of GO patients may contribute to changes in their ocular surfaces via diminished reactive oxygen species （ROS） depletion and adaptive immune responses.The completely absent of C3 in the GO patients＇ tears,may imply that an important inflammatory signaling pathway is affected,which needs further investigation.

Epstein-Barr virus-encoded small RNAs in idiopathic orbital inflammatory pseudotumor tissues: a comparative case series

AIM： To investigate the positive rate and types of cells that express Epstein-Barr virus-encoded small RNAs （EBERs） and to determine the distribution of EBER-expressing cells in idiopathic orbital inflammatory pseudotumor （IOIP） tissues. METHODS： We retrospectively examined 40 archived paraffin specimens from two teaching hospitals in Southern China between January 2007 and January 2015 that were pathologically determined to exhibit IOIP. Eleven concurrent paraffin specimens of thyroid-associated ophthalmopathy （TAO） composed the control group. In situ hybridization was performed to detect EBERs. Immunohistochemistry was employed to detect CD3, CD20, Vimentin, and smooth muscle actin （SMA）, and the positive rate, types of positive cells, and distribution and location of EBERs were evaluated. RESULTS： The positive expression rate of EBERs was 47.5% （19/40） in the IOIP group, which was significantly higher than that in the TAO group [0 （0/11）, P=-0.011]. in the IOIP group, the lymphocyte infiltrative subtype, fibrotic subtype, and mixed subtype exhibited EBER-positive rates of 57.1% （12121）, 12.5% （118）, and 54.5% （6/11）, respectively, and no significant differences were found between these subtypes （P=0.085）. Positive signals of EBERs were mainly present in medium-small lymphocytes between or around follicles and in the nuclei of activated immunoblasts （14/19）. CONCLUSION： The positive rate, types, and distribution of EBER-expressing cells in IOIP have been documented.These findings are conducive for a better understanding of the underlying mechanisms of Epstein-Barr virus infection in IOIP pathogenesis.

An overview of thyroid eye disease

Thyroid eye disease(also known as Graves’ophthalmopathy)is a complex orbital inflammatory disease,which can be sight threatening,debilitating and disfiguring.This overview discusses the epidemiology,risk factors,pathogenesis,presentation,ophthalmic clinical features,investigations and treatment of thyroid eye disease.