双色上转换纳米荧光探针在套细胞淋巴瘤细胞靶向成像中的应用

目的构建上转换纳米靶向探针,并探究其在特异性标记套细胞淋巴瘤细胞株中的应用价值,为体内套细胞淋巴瘤的靶向诊断提供理论依据和指导。方法采用H_2O_2对Na YF_4:Er^(3+)和Na YF_4:Yb^(3+),Tm^(3+)上转换纳米粒子表面进行氧化处理,使该粒子由最初的疏水性转变为亲水性;在NHS、EDC的作用下,亲水性的Na YF_4:Er^(3+)上转换纳米粒子与CD20单克隆抗体共价结合,Na YF_4:Yb^(3+),Tm^(3+)上转换纳米粒子与CD5单克隆抗体共价结合;CD20抗体-Na YF_4:Er^(3+)纳米粒子轭合物和CD5抗体-Na YF_4:Yb^(3+),Tm^(3+)纳米粒子轭合物的混合悬液与套细胞淋巴瘤细胞株室温下孵育2 h;CD20抗体-Na YF_4:Er^(3+)纳米粒子轭合物和Na YF_4:Yb^(3+),Tm^(3+)纳米粒子的混合悬液与套细胞淋巴瘤细胞株室温下孵育2 h;Na YF_4:Er^(3+)纳米粒子和CD5抗体-Na YF_4:Yb^(3+),Tm^(3+)纳米粒子轭合物的混合悬液与套细胞淋巴瘤细胞株室温下孵育2 h;未偶联有任何抗体的Na YF_4:Er^(3+)和Na YF_4:Yb^(3+),Tm^(3+)上转换纳米粒子的混合液与套细胞淋巴瘤细胞株室温下孵育2 h;使用配备有980 nm近红外激光的尼康Eclipse Ti-S倒置荧光显微镜对各组细胞进行上转换成像,观察细胞表面上转换荧光的强度。结果实验组细胞表面释放出蓝、绿双色的上转换纳米荧光,但通过观察3组对照实验,细胞表面却仅分别释放出单一的绿色荧光、蓝色荧光以及未见任何荧光。结论上转换纳米粒子与CD20或CD5抗体成功共价偶联,可以对套细胞淋巴瘤细胞株进行特异性标记、成像。

Influencing factors for peripheral and posterior lesions in mild non-proliferative diabetic retinopathy--the Kailuan Eye Study

AIM:To explore the influencing factors of diabetes type 2 patients with mild non-proliferative diabetic retinopathy(NPDR)in the Kailuan area of Tangshan,Hebei Province,China.METHODS:In this non-interventional,retrospective study,683 patients with type 2 diabetes were included in the Kailuan Diabetic Retinopathy Study involving participants with diabetes in the community-based longitudinal Kailuan Study.Based on the undilated ultrawide field(200°;UWF)images and partial dilated digital fundus images,the diabetic retinopathy(DR)of the surveyed population was graded.Interobserver agreement was estimated by using Cohen’s Kappa statistics.The main outcome indicators included gender,age,weight,height,body mass index,blood pressure,circumferences of neck,waist and hip,current smoking,levels of fasting plasma glucose(FPG),hypersensitive C-reactive protein,creatinine,and cholesterol,etc.According to different lesions’locations of patients with mild NPDR,logistic regression models were used to estimate the odds ratios(ORs)and their 95%Cls of each risk factor.RESULTS:The study group of 683 patients included 570 males and 113 females.The mean age of the patients was62.18±9.41 y.Compared with dilated fundus examinations,there was fair agreement with the level of DR identified on UWF images in 63.91%of eyes(k=0.369,95%CI,0.00-0.00).Detected by UWF images,there were 98 patients with mild NPDR having peripheral retinal lesions,35 patients with mild NPDR having posterior lesions,44 patients with mild NPDR whose lesions were detected both in and out the standard two fields area,and 336 patients with non obvious DR.Parameters that conferred a statistically significant increased risks for mild NPDR with having peripheral retinal lesions were neck circumstance(OR,1.124;95%CI,1.044-1.211),and with posterior lesions were FPG(OR,1.052;95%CI,1.007-1.099).CONCLUSION:UWF is an effectiveness means of DR screening.Moreover,it is necessary to evaluate peripheral diabetic retinal lesions which can help to estimate the severity of DR.The phenomenon that nonuniform and inhomogeneous distribution of DR lesions has been found.And the influencing factors in mild NPDR are differing by different lesions’locations.

Elevated VEGF-A & PLGF concentration in aqueous humor of patients with uveal melanoma following Iodine-125 plaque radiotherapy

AIM: To measure the concentration of vascular endothelial growth factor-A(VEGF-A), and placental growth factor(PLGF) in aqueous humor of uveal melanoma patients before and after Iodine-125 plaque therapy(IPT), determine the postoperative fluctuation and evaluate associated factors in vivo.METHODS: Participants were 18 Chinese patients with uveal melanoma who were elected to IPT. Undiluted aqueous humor samples were collected at Iodine plaque implant and removal time, then stored immediately at-80℃ until assayed. The concentration of VEGF-A, PLGF and other 7 cytokines comprising interleukin-2(IL-2), IL-8, IL-10, interferon(IFN)-γ, programmed death(PD)-1, transforming growth factor(TGF)-β1 and insulin-like growth factor(IGF)-1 in aqueous humor was measured using Raybiotech immunoassay kit, a high throughput strategy. The VEGF-A and PLGF levels were compared across preoperation and postoperation subgroups, as well as those of other 7 interleukins. Correlation and grouped analyses were conducted to determine the independent effects of clinical parameters and other cytokines on VEGF-A and PLGF concentration or fluctuation. This study set a self-control design.RESULTS: VEGF-A(P=0.038) and PLGF(P=0.026) were the only two increased cytokines after IPT. Preoperative and postoperative level of VEGF-A and PLGF(r=0.575, P=0.013;r=0.987, P<0.001) correlated with each other significantly. Level of VEGF-A(r=0.626, P=0.005;r=0.588, P=0.01) and PLGF(r=0.616, P=0.007;r=0.588, P=0.01) had positive correlation with tumor thickness consistently. Elevated VEGF-A or PLGF level were strong predictive factors of each other(P=0.007, OR=60.0). The elevated VEGF-A group showed a higher postoperative level of IFN-γ(P=0.005), IL-2(P<0.001) and IL-10(P=0.004) in aqueous humor. When the elevated PLGF group got similar results that a higher postoperative level of IFN-γ(P=0.007), IL-2(P<0.001) and IL-10(P=0.013) in aqueous humor. CONCLUSION: This study reveals that VEGF-A and PLGF in aqueous humor significantly increased with tumor thickness and radiation process in uveal melanoma patients. VEGF-A and PLGF may be crucial in uveal melanoma genesis and radiotherapy reactions. Immune mediators comprised IFN-γ, IL-2 and IL-10 could play roles in the link between inflammation and angiogenesis in uveal melanoma when exposed to radiotherapy.

Machine learning models for outcome prediction of Chinese uveal melanoma patients: A 15-year follow-up study

Dear Editor Uveal melanoma(UM)is the most frequent primary malig-nant intraocular tumor in adults with an estimated inci-dence of 4-5 per million per year in western countries[1].About 50%of UM patients eventually develop metas-tasis.In a previous study,the prognosis of Chinese UM was mainly correlated with visual clinical features and gene sequencing results.Models designed to predict UM prognosis have been previously described[2],but these studies were based on Caucasians,not Chinese.