Accurate collection and preservation of vitreous and retina-related tissue specimens is the basis for clinical diagnosis and rigorous basic research. The clinical uses of vitreous specimens include microbial culture, cytological detection, detection of degenerative diseases, PCR analysis, and cytological detection of cell morphology. The experimental research uses include DNA gene analysis, protein quantitative analysis, metabolite examination, RNA content quantitative analysis, cytokine determination and so on. Retinal specimens collecting was mainly used for PCR analysis of retinal proliferative membrane, immunohistochemical staining, immunofluorescence examination, microvascular density evaluation, cell isolation and culture, etc. Understanding the collection of vitreoretinal surgical specimens and the application of relevant detection techniques and materials can provide a more comprehensive idea for the diagnosis of vitreoretinal diseases and a broader reference for the related basic research.
ObjectiveTo observe the changes in refractive status of eyes with idiopathic macular hole (IMH) after vitrectomy and phacoemulsification and IOL implantation (combined surgery).MethodsA retrospective clinical study. From January 2016 to June 2019, 51patients (56 eyes) of IMH who underwent combined surgery at the Tianjin Medical University Eye Hospital. were included in the study. Among them, there were 17 males and 34 females with the average age of 66.79±4.33 years. All the affected eyes underwent BCVA, retinoscopy and axial length (AL) measurement. The IOL power was calculated according to the SRK-T formula and the refractive power (predicted value) was predicted. The average BCVA of the affected eye was 0.20±0.13. The average anterior chamber depth was 2.89±0.28 mm. The average △corneal astigmatism was 0.73±0.43 D, the average AL was 22.92±0.70 mm, the average predicted refractive power was 0.10±0.66 D. All the affected eyes underwent standard transciliary flat part three-channel 25G combined surgery. Six months after the operation, the actual value (actual value) of the diopter after the operation was measured with the same equipment and method before the operation. Paired t test was used to compare the difference between the predicted value and the actual value.ResultsSix months after the operation, the actual value of the refractive power was -0.19±0.64 D. Compared with the pre-operative refractive power, the difference was not statistically significant (t=1.665, P=0.102). The difference between the actual value and the predicted value was －0.33±0.81 D.ConclusionsThe refractive status of the IMH eye undergoes myopia drift after combined surgery. The preoperative IOL power budget can be appropriately reserved for +0.3 D hyperopia.
ObjectiveTo observe the changes of follistatin-like protein 1 (FSTL1) in serum of patients with proliferative diabetic retinopathy (PDR).MethodsTwenty PDR patients confirmed by clinical examination and 20 normal people were included in the study. Human retinal vascular endothelial cells (HRCEC) were divided into HRCEC blank control group, 3 h hypoxia group, 6 h hypoxia group. Human umbilical vein endothelial cell (HUVEC) were divided into HUVEC blank control group, 3h hypoxia group, 6h hypoxia group. Real-time quantitative PCR (RT-PCR) and ELISA were used to determine the expression of FSTL1, TGF-β, VEGF, connective tissue growth factor (CTGF) mRNA and protein in peripheral blood and cells of all groups from all subjects.ResultsThe expressions of FSTL1, TGF-β1, CTGF, VEGF mRNA in blood samples of patients with PDR were 1.79±0.58, 0.97±0.21, 1.85±0.69 and 1.38±0.44. The expressions of FSTL1, TGF-β1 protein were 1.19±0.50, 0.71±0.24 ng/ml and 734.03±116.45, 649.36±44.23 ng/L. Compared with normal people, the differences were statistically significant (tmRNA=0.90, 0.21, 2.85, 1.77; P=0.00, 0.00, 0.04, 0.02. tprotein=1.88, 7.68; P=0.00, 0.02). The cell viability of HRCEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.66±0.05 and 0.64±0.04, respectively. Compared with the blank control group, the difference was statistically significant (F=13.02, P=0.00). The cell viability of HUVEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.63±0.06 and 0.68±0.06, respectively. Compared with the blank control group, the difference was statistically significant (F=26.52, P=0.00). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HRCEC blank control group and 3 h hypoxia group, the differences were statistically significant (F=14.75, 44.93, 85.54, 6.23; P=0.01, 0.00, 0.00, 0.03). Compared with the HRCEC blank control and 3 h hypoxia group, the expressions of FSTL1 and TGF-β1 protein were statistically significant (P＜0.05). There was a statistically significant difference in TGF-β1 protein expression in the hypoxic 6 h group (P=0.03) and no significant difference in FSTL1 protein expression (P=0.68). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HUVEC blank control group and 3h hypoxia group, the differences were statistically significant (F=19.08, 25.12, 22.89, 13.07; P=0.00, 0.00, 0.00, 0.01). Immunofluorescence staining results showed that FSTL1, TGF-β1, CTGF, and VEGF proteins were positively expressed in cells in the 3h hypoxia and 6h hypoxia groups.ConclusionThe expression of FSTL1 gene and protein in serum of PDR patients was significantly higher than that of normal people.