The application of wide-field laser ophthalmoscopy in fundus examination before myopic refractive surgery.

BACKGROUND: To evaluate wide-field laser ophthalmoscopy (Optomap 200Tx) for screening retinal lesions before myopic refractive surgery. METHODS: Seventy-eight eyes of 78 consecutive refractive surgery candidates were included in this study. All subjects underwent Optomap 200Tx, mydriatic slit-lamp lens examination and the Goldmann three-mirror contact lens examination, which was considered as the reference method for determining retinal lesions. RESULTS: Forty of 78 eyes had retinal lesions (51.28%) and three eyes had retinal breaks (3.85%), which were diagnosed by the Goldmann three-mirror contact lens examination. Compared to the Goldmann three-mirror contact lens examination, the detection rate with the Optomap 200Tx was 91.73%% for retinal lesions, while the detection rate of mydriatic slit-lamp lens exams was 81.20%. There were no statistically significant differences among the three methods used for the diagnoses of myopic conus, tessellation and retinal breaks(all p > 0.05). For peripheral retinal lesions, the detection rate of the Optomap 200Tx examinations were similar to the Goldmann three-mirror contact lens exams (all p > 0.05), but were higher than the results of slit-lamp lens examinations (all p < 0.05). Regarding the vitreoretinal adhesions, the Goldmann three-mirror contact lens examinations had higher detection rates than did the Optomap 200Tx examinations (p = 0.031). CONCLUSIONS: The Optomap 200Tx examinations is a convenient and feasible method to determine fundus pathological changes in myopic patients, especially for patients who can not endure pupil dilation. In order to avoid misdiagnosis of peripheral retinal lesions, Goldmann three-mirror contact lens examination is needed.

Vitreous proteomic analysis of idiopathic epiretinal membranes.

To understand the molecular mechanisms of idiopathic epiretinal membranes (iERMs), the vitreous proteomes of patients with iERMs were investigated. The vitreous proteome in patients with iERMs (n = 8) and donor samples (n = 8) was analysed using reversed phase high-performance liquid chromatography (RP-HPLC) coupled with electrospray ionization tandem mass spectrometry (ESI-MS/MS) and GeneGo Metacore™. This research followed the tenets of the Declaration of Helsinki for the use of human subjects. In this current study, 226 significant changes in protein abundance (abundance ratio >2, p < 0.01) were identified in the vitreous proteome of iERM patients compared to normal control vitreous, including 122 proteins that were present at lower levels and 104 proteins that were present at higher levels. In the iERM vitreous samples, complement components, inflammation-related proteins and matrix metalloproteinase were present at higher levels, while normal cytoskeleton proteins were present at lower levels. The top GeneGo pathway was "immune response", the top process network was "inflammation", and the top KEGG pathway was "coagulation cascades". The essential 2-node proteins of the network were estrogen receptor 1 (ESR1) and p300. Among those found at higher levels, ubiquitin-conjugating enzyme E2O (UBE2O) and complement C4A (C4A) were the most abundant proteins, and could be detected in each of the iERM vitreous samples. It can be concluded that iERMs are a complicated pathological process involving inflammation, immune response, and cytoskeleton remolding. UBE2O and C4A may be candidate biomarkers for iERMs.

Differentiating vitreous proteomes in proliferative diabetic retinopathy using high-performance liquid chromatography coupled to tandem mass spectrometry.

Proliferative diabetic retinopathy (PDR) is a serious microangiopathic complication of diabetes mellitus and a major cause of blindness in working-age adults. Diabetes-induced alterations in the vitreous protein composition in diabetic patients with PDR may be responsible for the presence of PDR. Therefore, we performed a comprehensive proteomic analysis and compared the protein profiles of vitreous humor from type 2 diabetic patients with PDR (n = 8) and that from normal human eyes donated for corneal transplant (n = 8). Using reversed phase high-performance liquid chromatography (RP-HPLC) coupled to electrospray Ionization tandem mass spectrometry (ESI-MS/MS), we identified 96 significant differentially expressed proteins (abundance ratio > 1.5, p < 0.05), including 37 and 59 proteins up- and downregulated in PDR vitreous compared with the control, respectively. Biological pathway analysis revealed 44 proteins involved in 56 biological pathways; among them, the most remarkable pathways differentially represented between PDR and normal vitreous were the glycolysis/gluconeogenesis, complement and coagulation cascades, gap junction, and phagosome pathways. The differential expressions of angiopoietin-related protein 6, apolipoprotein A-I, estrogen receptor alpha, and tubulin were confirmed by western blot analysis. These data provide insight into the molecular events possibly involved in the pathogenesis of PDR and widen the scope of potential avenues for new therapies for PDR.

Berberine inhibits the invasion and metastasis of nasopharyngeal carcinoma cells through Ezrin phosphorylation.

OBJECTIVE: To determine the molecular mechanism of berberine (BBR) inhibiting the metastasis and invasion of nasopharyngeal carcinoma 5-8F cells, and identify whether berberine suppresses the tumor-invasive action through inhibiting Ezrin or phosphate-Ezrin. METHODS: The non-cytotoxic concentration of berberine was detected by MTT assay. Filopodia formation of 5-8F cells was observed by electron microscope. The invasion and motility of 5-8F cells with berberine treatment were measured with Trans-well assay. Western blot was used to investigate the Ezrin and phos-Ezrin expression in 5-8F cells treated by berberine. pcDNA3.1-Ezrin and pcDNA3.1-Ezrin M were transfected into 6-10B cells. The inhibitory effect of berberine on the motility and invasion of 6-10B-pcDNA3.1-Ezrin and 6-10B-pcDNA3.1-Ezrin M was detected, respectively. RESULTS: Berberine non-cytotoxic concentration was 0-40 µmol/L. After being treated by berberine, filopodia of 5-8F cells obviously reduced, and the permeating artificial basement membrane cells largely decreased in both time- and concentration-dependent manner. There was significant difference compared with the control group (P<0.05). Berberine suppressed the phos-Ezrin expression of 5-8F cells in both time- and concentration-dependent manner (P<0.05), but the effect of berberine was weaker on 6-10B-pcDNA3.1-Ezrin M than on 6-10B-pcDNA3.1-Ezrin. CONCLUSION: Berberine inhibits nasopharyngeal carcinoma cell invasion through inhibiting phos-Ezrin expression and filopodia formation.

Novel potential markers of nasopharyngeal carcinoma for diagnosis and therapy.

OBJECTIVE: To search for markers of nasopharyngeal carcinoma (NPC) for diagnosis. DESIGN AND METHODS: Using gas chromatography and mass spectrometry, we evaluated 51 serum metabolites in 49 NPC, 37 throat cancer patients and 40 healthy controls. High metabolites were selected and confirmed in NPC tissues. Sensitivity and specificity were appraised for 53 NPC diagnoses. RESULTS: Metabolic profiling revealed that kynurenine, N-acetylglucosaminylamine, N-acetylglucosamine and hydroxyphenylpyruvate increased in NPC patient sera. Their sensitivity and specificity were respectively 79% and 71%, 78% and 69%, 83% and 68%, 84% and 73% for NPC diagnosis. These increases were confirmed in NPC cells. Four metabolites gradually increased from stage I to stage III. After radiotherapy, four metabolites decreased gradually, and tended to a normal level, and were associated with rate of tumor reduction. CONCLUSION: The results reveal that kynurenine, N-acetylglucosaminylamine, N-acetylglucosamine and hydroxyphenylpyruvate are potentially markers of NPC for diagnosis and therapy.

Berberine inhibits metastasis of nasopharyngeal carcinoma 5-8F cells by targeting Rho kinase-mediated Ezrin phosphorylation at threonine 567.

Ezrin is highly expressed in metastatic tumors and is involved in filopodia formation as well as promotion of tumor metastasis. Thus, Ezrin may serve as a potential target for anti-metastatic therapy. This study demonstrates that berberine reduces filopodia formation of a nasopharyngeal carcinoma (NPC) cell line, 5-8F, at non-cytotoxic concentrations. Furthermore, invasion and motility of 5-8F cells are decreased in a dose- and time-dependent manner, resulting in 73.0% invasion and 67.0% motility inhibition at 20 mum. The inhibitory effects of berberine on 5-8F cell metastasis were further confirmed in a mouse model of metastasis. Berberine treatment in vivo resulted in a 51.1% inhibition of tumor metastasis to the lymph nodes and decreased Ezrin phosphorylation at threonine 567 in metastatic samples. Berberine suppressed the presence of phosphorylated Ezrin (phospho-Ezrin) in a dose- and time-dependent manner but had no effect on total Ezrin protein expression at non-cytotoxic concentrations. Furthermore, the inhibitory effects of berberine on phospho-Ezrin were dependent on the suppression of Rho kinase activity. Reduction of Ezrin phosphorylation at Thr(567) by berberine was associated with its inhibitory effect on filopodia formation in 5-8F cells. However, berberine did not effectively inhibit the motility and invasion of NPC cells containing Ezrin Thr(567) mutants. These results confirm that berberine inhibits Ezrin phosphorylation at Thr(567). Nonetheless, berberine reduces motility and invasion of cells and inhibits tumor metastasis. The reduction of Rho kinase-mediated Ezrin phosphorylation mediated by berberine may be a novel anti-metastatic pathway in NPC 5-8F cells.

HSP70 and mucin 5B: novel protein targets of N,N'-dinitrosopiperazine-induced nasopharyngeal tumorigenesis.

N,N'-Dinitrosopiperazine (DNP) induces nasopharyngeal carcinoma (NPC) and shows organ specificity to the nasopharyngeal epithelium. To investigate its mechanism, the rat NPC model was induced using DNP. Rat NPC and normal nasopharyngeal cells were obtained from the NPC model using laser capture. The total proteins from these cell samples were separated with two-dimension polyacrylamide gel electrophoresis techniques, and highly expressed proteins (> five-fold) were analyzed using matrix-assisted laser desorption/ionization time of flight and bioinformatics. The results showed that HSP70 and mucin 5B expression increased not only in rat NPC but also in atypical hyperplasia nasopharyngeal tissues, a precancer stage of NPC. High-expression of heat shock protein 70 (HSP70) and mucin 5B was further supported by western blot analysis. The immunofluorescence and western-blotting studies further showed that DNP induced the expression of HSP70 and mucin 5B in a dosage-dependent manner in normal nasopharyngeal epithelia cells. Our data indicate that DNP triggers over-expression of HSP70 and mucin 5B, and is involved in nasopharyngeal tumorigenesis. HSP70 and mucin 5B may be important targets in nasopharyngeal tumorigenesis induced by DNP.