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BACKGROUND: Macular edema (ME) is a major complication of retinal disease with multiple mechanisms involved in its development. This study aimed to investigate the metabolite profile of aqueous humor (AH) in patients with ME of different etiologies and identify potential metabolite biomarkers for early diagnosis of ME. METHODS: Samples of AH were collected from 60 patients with ME and 20 age- and sex-matched controls and analyzed by liquid chromatography-mass spectrometry (LC/MS)-based metabolomics. A series of univariate and multivariate statistical analyses were performed to identify differential metabolites and enriched metabolite pathways. RESULTS: The metabolic profile of AH differed significantly between ME patients and healthy controls, and differentially expressed metabolites were identified. Pathway analysis revealed that these differentially expressed metabolites are mainly involved in lipid metabolism and amino acid metabolism. Moreover, significant differences were identified in the metabolic composition of AH from patients with ME due to different retinal diseases including age-related macular degeneration (AMD-ME), diabetic retinopathy (DME) and branch retinal vein occlusion (BRVO-ME). In total, 39 and 79 etiology-specific altered metabolites were identified for AMD-ME and DME, respectively. Finally, an AH-derived machine learning-based diagnostic model was developed and successfully validated in the test cohort with an area under the receiver operating characteristic (ROC) curve of 0.79 for AMD-ME, 0.94 for DME and 0.77 for BRVO-ME. CONCLUSIONS: Our study illustrates the potential underlying metabolic basis of AH of different etiologies across ME populations. We also identify AH-derived metabolite biomarkers that may improve the differential diagnosis and treatment stratification of ME patients with different etiologies.
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BACKGROUND: The impact and potential molecular mechanisms of SST in the occurrence and development of GC have not been determined. MATERIALS AND METHODS: Two pairs of sgRNA and reporter were designed according to targeting sequence of SST gene for double-nicking. Plasmids were transfected into 293T for selecting sgRNA with higher cutting efficiency. The subline which has knocked-out SST gene were selected by FACS and verified by sequencing and expression level. Moreover, the migration and invasion ability was evaluated by wound healing and transwell after knocking out SST. Besides, the protein expression of SEMA5A and KLF2 were observed by Western blotting and LSCM. Last, we detected the expression levels of SST, SEMA5A, and KLF2 in GC tissues by Western blotting. RESULTS: The results revealed that the new subline 1E9, which had knocked out SST gene, was established by CRISPR/Cas9. In addition, the knockout of SST in GC cells markedly increased migration and invasion ability. The results also demonstrated that the knockout of SST increased the expression of SEMA5A and KLF2. The expression level of SST was decreased in GC tissues, and its decrease was associated with overexpression of SEMA5A and KLF2. CONCLUSION: SST plays an inhibitory role in the migration and invasion of GC cell BGC823. The protein expression levels of SEMA5A and KLF2 were enhanced in GC cells and tissues lacking SST expression.
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UNLABELLED: Humic acid (HA), a fluorescent allomelanin, has been implicated as an etiological agent of Blackfoot disease (BFD), a peripheral vascular disease prevailing in the southwest of Taiwan. Clinical and pathological studies reveal that it is similar to atherosclerosis. In this report, the effect of HA on human neutrophils is studied because prolonged and enhanced activation of neutrophils adhered on endothelium may damage the endothelium and initiate the process of thrombosis and vasculitis. METHODS: Neutrophils, treated with various concentrations of HA, were added to culture plates, cultured human umbilical vein endothelial cells (HUVECs), or human umbilical vein endothelium tissue culture for 15 or 30 min. The adhesion of neutrophils was measured qualitatively and quantitatively. The mechanism of neutrophil activation was studied with free radical production and various kinase measurements and their activities' assays. RESULTS: HA was shown to enhance, in a dose-dependent manner, the adhesion of neutrophils on the culture plates, cultured human umbilical vein endothelial cells, and human umbilical vein endothelium tissue culture. The adhesion-enhancing ability of HA is elicited through activation of ERK, P38 mitogen-activated kinase (P38MAPK), and phosphoinositide 3 kinase (PI3K) in neutrophils. HA also induces the NF-kappaB activation in neutrophils. CONCLUSION: HA treatment markedly enhanced adhesion and superoxide radical production of neutrophils, the characteristics of activated neutrophils; and all these stimulation effects were blocked by several kinase inhibitors, reflecting the involvement of the ERK, P38MAPK, and PI3K on the activation of neutrophils. The induction of NF-kappaB implied that the consequence of neutrophil activation by HA were similar to other stimulants. The prolonged neutrophil activation will further damage endothelium cell and cause thrombosis, vaculitis, as well as arteriosclerosis. This may partially explain why HA consumption will cause BFD.
