Objective To investigate the changes of macular microcirculation and aqueous humor cytokine expression in patients with diabetic macular edema(DME)after anti-vascular endothelial growth factor(VEGF)treatment,and analyze the relationship with efficacy.Methods A total of 62 patients(91 eyes)with DME who were treated in the First Hospital of Nanchang from October 2021 to August 2023 were selected and treated with intravitreal injection of conbercept.According to the reduction of central macular thickness(CMT),they were divided into efficacy significant group(CMT reduction≥100μm,59 eyes)and non-efficacy significant group(CMT reduction＜100μm or increase,32 eyes).The changes of CMT,vessel density(VD)of superficial capillary plexus(SCP),fovea avascular area(FAZ),VEGF,interleuki(IL)-6,IL-8,and IL-10 after anti-VEGF treatment were analyzed.Receiver operating characteristic(ROC)curve was used to evaluate the predictive value of each index.Results Before treatment,the levels of VEGF and IL-10 in aqueous humor in efficacy significant group were significantly higher than those in non-efficacy significant group,and the level of IL-8 was significantly lower than that in non-efficacy significant group(P＜0.05).After treatment,levels of VEGF,IL-6,IL-8 and IL-10 in aqueous humor in both groups were significantly lower than before treatment(P＜0.05).The levels of VEGF,IL-6 and IL-8 in aqueous humor in efficacy significant group were significantly lower than those in non-efficacy significant group,and the level of IL-10 was significantly higher than that in non-efficacy significant group(P＜0.05).Before and after anti-VEGF treatment,there were no significant changes in FAZ area and SCP-VD in both groups(P＞0.05).Correlation analysis showed that VEGF(r=0.571,P＜0.001)and IL-10(r=0.382,P=0.008)in aqueous humor at baseline were positively correlated with CMT reduction,IL-8 was negatively correlated with CMT reduction(r=-0.689,P＜0.001).IL-6,FAZ area and SCP-VD were not correlated with CMT reduction(P＞0.05).Cytokine levels were not correlated with FAZ area and SCP-VD(P＞0.05).ROC curve results showed that area under the curve of IL-8,VEGF and IL-10 at baseline predicting anti-VEGF efficacy were 0.825,0.813 and 0.676,respectively.Conclusion The levels of VEGF,IL-8,and IL-10 in aqueous humor at baseline in DME patients were correlated with anti-VEGF efficacy and could predict the efficacy of anti-VEGF.

Objective : To investigate the effects of fukutin ( FKTN) which is a member of ribitol⁃5⁃phosphate transferases on HeLa cell cycle, apoptosis, migration and the secretion of α⁃dystroglycan(α⁃DG) . Methods : The plasmid pcDNA3. 1⁃α⁃DAG⁃HA was constructed. HeLa cells were transfected with plasmid pcDNA3. 1⁃FKTN ⁃ 3xFlag, then the total protein was extracted for Western blot to determine the expression level of FKTN. Cell cycle and apoptosis were measured by flow cytometry after the overexpression of FKTN. Following the overexpression of FKTN, wound⁃healing assay was performed to detect the cell migration rate, as the same time wheat germ agglutinin⁃agarose (WGA⁃agarose) was used to enrich α⁃DG in the cell cultures, then α⁃DG was detected by Western blot. Results : The eukaryotic expression plasmid pcDNA3. 1⁃α⁃DAG⁃HA was constructed successfully. FKTN could be overexpressed in HeLa cells. After the overexpression of FKTN,the percentage of S phase of cell cycle in the experimental group decreased (P < 0. 001) and apoptosis rate unchanged (P > 0. 05) when compared with the control group. There was no change in the cell migration rate of experimental group (P > 0. 05), but after the overexpression of FKTN, secretion of α⁃DG decreased when compared with control group. Conclusion Overexpressing FKTN arrests cell cycle and inhibits the secretion of α⁃DG in HeLa cells. Apoptosis and cell migration of HeLa cells are not affected by the overexpression of FKTN.

Objective : To investigate the interaction between intracellular chloride ion protein 1(CLIC1) and activated protein kinase C receptor 1 ( RACK1) . Methods : Plasmids pcDNA3. 1⁃RACK1⁃HA and/or pcDNA3. 1 ⁃CLIC1⁃FLAG were transfected into HEK 293T cells, and pcDNA3. 1⁃RACK1⁃HA and/or pcDNA3. 1⁃CLIC1⁃FLAG were transfected into COS7. GST⁃pulldown and immunoprecipitation assays were performed to determine the interaction between CLIC1 and RACK1 in vivo and in vitro. The co⁃localization of CLIC1 and RACK1 was observed by indirect immunofluorescence assay. Results : Western blot confirmed that CLIC1 and RACK1 could be highly expressed in HEK 293T cells. GST⁃pulldown showed that RACK1 bound CLIC1 in vitro, and immunoprecipitation showed that CLIC1 and RACK1 interacted in vivo. Furthermore, indirect immunofluorescence assay showed CLIC1 co⁃localized with RACK1 . Conclusion Human CLIC1 protein interacts with RACK1 in vitro and in vivo.