Single cell RNA sequencing technique provides a strong technical support for the analysis of cell heterogeneity in biological tissues, and has been widely used in biomedical research. In recent years, considerable scRNA-seq data have been accumulated in the research of ocular fundus diseases. The ocular fundus is abundant for the network of vessel and neuron, which leads to the complicated pathogenesis of fundus diseases. Through single cell RNA sequencing technique, the expression of thousands of genes of certain cell types or even subtypes can be obtained in the disease environment. Single cell RNA sequencing technique accurately reveals the pathogenic cell types and pathogenic mechanisms of ocular fundus diseases such as neovascular retinopathy, which provides a theoretical basis for the birth of new diagnosis and treatment targets. The construction of multi-omics single-cell database of ocular fundus diseases will enable high-quality data to be further explored and provide an analysis platform for ophthalmic researchers.

Objective:To evaluate the risk factors for the complexity and surgical prognosis in patients with proliferative diabetic retinopathy (PDR).Methods:A historical cohort study of the CONCEPT trial, including 97 patients (97 eyes) who were diagnosed with PDR and requiring three-channel 23-gauge transconjunctival pars plana vitrectomy (PPV) from June 2017 to January 2018 at the First Affiliated Hospital of Nanjing Medical University.All patients received preoperative intravitreal injection of 0.5 mg conbercpet.Based on the PDR complexity score, patients were divided into >3 group or ≤3 group, and the systematic risk factors were compared between the two groups.The influence of sex, age, hypertension, renal insufficiency, duration of diabetes mellitus, and hemoglobin A1c level on the PDR complexity score was evaluated by multivariate logistic regression analysis.Based on age, patients were divided into <45 years group, 45-<60 years group, and ≥60 years group, and the differences in mean operative time, incidence of intraoperative hemorrhage, surgically induced lacrimation and silicone oil filling, and incidence of hemorrhage on color fundus photos and macular edema by optical coherence tomography at postoperative months 1 and 6 were analyzed among different age groups.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of The First Affiliated Hospital of Nanjing Medical University (No.2017-SR-283).Written informed consent was obtained from each subject.Results:The age of patients with PDR complexity score >3 was 46.5(36.0, 51.8) years, which was less than 54.0(45.5, 61.5) years for PDR complexity score ≤3, and the difference was statistically significant ( Z=1.835, P=0.002).Among the factors predicting PDR complexity score >3, logistic regression analysis indicated that only age was statistically significant ( P=0.005).For each 1-year increase in age, the risk of PDR complexity score >3 would increase by 7.4%( OR: 0.929, 95% CI: 0.883-0.977).Among the systemic factors, there were significant differences in age, history of diabetes, proportion of patients with hypertension and renal insufficiency among the three age groups (all at P<0.05).Among the ocular factors, there were significant differences in the proportion of patients with history of retinal laser treatment, fibrovascular membrane and complexity score >3 among the three groups (all at P<0.05).The proportion of patients with fibrovascular membrane and complexity score >3 in the <45 years group was significantly higher than that in the 45-<60 and ≥60 years groups (all at P<0.05).There were significant differences in the proportion of patients with intraoperative bleeding and silicone oil filling in the three age groups (all at P<0.017).The proportion of intraoperative bleeding and silicone oil filling in <45 years group was significantly higher than that in 45-<60 and ≥60 years groups (all at P<0.05).The macular edema on postoperative month 1 in the <45 years old group was significantly higher than that in the 45-<60 and ≥60 years groups (both at P<0.05). Conclusions:Among systemic factors, age has a significant impact on the increased PDR complexity and contributes to the poor prognosis of patients.There is a higher percentage of intraoperative complications and early postoperative macular edema in patients in the younger age group compared to the older age group.

Objective:To apply the multi-modal deep learning model to automatically classify the ultra-widefield fluorescein angiography (UWFA) images of diabetic retinopathy (DR).Methods:A retrospective study. From 2015 to 2020, 798 images of 297 DR patients with 399 eyes who were admitted to Eye Center of Renmin Hospital of Wuhan University and were examined by UWFA were used as the training set and test set of the model. Among them, 119, 171, and 109 eyes had no retinopathy, non-proliferative DR (NPDR), and proliferative DR (PDR), respectively. Localization and assessment of fluorescein leakage and non-perfusion regions in early and late orthotopic images of UWFA in DR-affected eyes by jointly optimizing CycleGAN and a convolutional neural network (CNN) classifier, an image-level supervised deep learning model. The abnormal images with lesions were converted into normal images with lesions removed using the improved CycleGAN, and the difference images containing the lesion areas were obtained; the difference images were classified by the CNN classifier to obtain the prediction results. A five-fold cross-test was used to evaluate the classification accuracy of the model. Quantitative analysis of the marker area displayed by the differential images was performed to observe the correlation between the ischemia index and leakage index and the severity of DR.Results:The generated fake normal image basically removed all the lesion areas while retaining the normal vascular structure; the difference images intuitively revealed the distribution of biomarkers; the heat icon showed the leakage area, and the location was basically the same as the lesion area in the original image. The results of the five-fold cross-check showed that the average classification accuracy of the model was 0.983. Further quantitative analysis of the marker area showed that the ischemia index and leakage index were significantly positively correlated with the severity of DR ( β=6.088, 10.850; P<0.001). Conclusion:The constructed multimodal joint optimization model can accurately classify NPDR and PDR and precisely locate potential biomarkers.

Objective:To analyze the changes of subfoveal choroidal thickness (SFCT) and choroidal volume in type 2 diabetes patients with diabetic retinopathy (DR) through automated choroid segmentation.Methods:A cross-sectional study was conducted.Sixty-four patients (88 eyes) with type 2 diabetes diagnosed in The First Affiliated Hospital with Nanjing Medical University from May 2016 to May 2017 were enrolled, including 27 males (32 eyes) and 37 females (56 eyes), with an average age of (62.7±7.4) years and an average diabetes duration of (13.7±7.2) years.Best corrected visual acuity, slit lamp ophthalmoscopy, indirect ophthalmoscopy and spectral domain optical coherence tomography (SD-OCT) examination were carried out among all the patients.The patients were divided into non-DR (NDR) group (28 eyes), nonproliferative DR (NPDR) group (36 eyes) and proliferative DR (PDR) group (24 eyes) according to the Early Treatment Diabetic Retinopathy Study.Nineteen age-matched normal subjects (32 eyes) were enrolled as normal control group.SFCT and choroidal volume were measured through automated choroid segmentation based on SD-OCT with enhanced depth imaging.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of The First Affiliated Hospital with Nanjing Medical University (No.2017-SR-176).Results:Mean SFCT and choroidal volume of normal control group, NDR group, NPDR group and PDR group were (277.8±45.3)μm and (8.8±1.0)mm 3, (220.9±38.7)μm and (7.6±1.3)mm 3, (231.8±26.7)μm and (8.2±0.9)mm 3, (252.0±42.2)μm and (8.7±1.0)mm 3, respectively, with significant overall differences among the groups ( F=14.615, P<0.001; F=7.711, P<0.001). Mean SFCT and choroidal volume were significantly smaller in NDR and NPDR group than those in normal control group (both at P<0.05). Mean SFCT and choroidal volume of PDR group was greater than those of NDR group (both at P<0.05). Conclusions:SFCT and choroidal volume decrease during the early course of diabetes and increase significantly as DR worsens from NDR to PDR.

Objective To compare the transparency efficiency of six different optical clearing method on the rat brain tissues. Methods Brain tissue blocks of 14 SD rats were processed with iDISCO, SeeDB, CUBIC, SCALEVIEW-A2,CLARITY-CUBIC, Passive-CLARITY clearing method, respectively. Results The gray value of PBS group was 13.031 ± 0.586,that of iDISCO,SeeDB,CUBIC,SCALEVIEW-A2,CLARITY-CUBIC,passive-CLARITY clearing were 6.447 ± 0.574,11.690 ± 0.909,2.318 ± 0.986,8.118 ± 1.026,8.591 ± 0.384,4.198 ± 0.182, respectively. Except the SeeDB group(P=0.185),the rest groups showed significant differences compared with the PBS group(P< 0.01), and there were significant differences between CUBIC and other groups(P < 0.01). After the clearing treatment, the changes of tissue area ratio in the iDISCO, SeeDB, CUBIC, SCALEVIEW-A2, CLARITY-CUBIC, Passive-CLARITY method were(-30.02 ± 2.39)%,(19.74 ± 4.09)%,(14.7 ± 3.92)%,(10.7 ± 5.55)%,(23.01 ± 4.19)%,(66.51 ± 5.68)%,respectively. Each group showed a significant difference compared with the groups iDISCO and the Passive-CLARITY,P< 0.01. Conclusions Except the SeeDB method,all the clearing methods can achieve a transparent effect, while CUBIC is better than the other groups applied for rat brain tissues. The tissue block volume is shrunken after iDISCO clearing,and expanded after Passive-CLARITY processing.

Replacement of diseased retinal pigment epithelium (RPE) cells with healthy RPE cells by transplantation is one option to treat several retinal degenerative diseases including age-related macular degeneration,which are caused by RPE loss and dysfunction.A cellular scaffold as a carrier for transplanted cells,may hold immense promise for facilitating cell migration and promoting the integration of RPE ceils into the host environment.Scaffolds can be prepared from a variety of natural and synthetic materials.Strategies,such as surface modification and structure adjustment,can improve the biomimetic properties of the scaffolds,optimize cell attachment and cellular function following transplantation and lay a foundation of clinical application in the future.

Objective To investigate the difference of vacuum sealing drainage (VSD) on the effect of one-stage skin graft and second-stage sugery after scar release.Methods A total of 42 patients who wanted to undergo scar release and skin graft was randomly divided to control group (n =21) and VSD treatment group (n =21).The control group implemented skin graft immdiately after scar release while VSD treatment group were treated with VSD for 3 days after scar release and then implemented skin graft.The rate of subcutaneous blood stasis and the survival rate of skin graft were observed at 7 days after skin graft.The condition of grafted skin contracture and hyperplasia after half a year was also observed.Results The incidcnce of subcutaneous blood stasis was significantly lower in the VSD group than that in the control group (P ＜ 0.05).The survival rate of skin grafts was significantly higher in the VSD group than that of the control group (P ＜ 0.05).The score of Vancouver scar was significantly lower in the VSD group than that in the control group (P ＜ 0.05).Conclusions VSD treatment of second-stage sugery after scar release can reduce the occurrence of subcutaneous blood stasis,promotc skin graft survival,reduce postoperative skin graft contracture and improve the prognosis of patients compared to one-stage skin graft.

Objective To investigate the influence of abnormal lipid metabolism on prognosis and quality of life in patients with severe COPD. Methods 134 patients with severe COPD in our hospital from September 2013 to January 2015 were selected as research objects, and their blood lipid levels were detected. The patients were divided into the normal group (n=61) and the low level group (n=73) according to the test results of lipid levels. 1-year mortality rate was statistically analyzed in both groups , and the WHOQOL-BREF health survey profile was applied to assess quality of life before and after treatment. Results 1-year survival rate in the normal group was 90.16%(55/61), which was higher than 73.97%(54/73) in the low level group (χ2=5.741, P=0.017). As compared with the normal group, the scores on physiological function, environment, social relationship, mental state, spirit / religious / belief, and total quality of lifewere markedly lower in the low level group after treatment (P < 0.05). The correlation analysis showed that plasma levels of TC, TG, and HDL-C were negatively correlated with 1-year mortality rate in patients with severe COPD (P < 0.05) but positively related with all dimensions in quality of life; plasma HDL-C level was positively related with 1-year mortality rate (P < 0.05), but negatively related with all dimensions in quality of life in patients with severe COPD (P < 0.05). Conclusions For patients with severe COPD, abnormal lipid metabolism is closely related with prognosis and quality of life. We should pay attention to nutritional support and control of abnormal lipid metabolism in those patients , leading to improvement of prognosis and quality of life.

Based on the pathogenic mechanisms of age-related macular degeneration (AMD),tremendous preclinical and clinical trials have demonstrated that cell transplantation which aim to replace impaired retinal pigment epithelium (RPE) with healthy RPE cells is a promising approach to treat AMD.So far,choices of cell sources mainly are autologous RPE,iris pigment epithelium,fetal RPE,human embryonic stem cell-derived RPE and human induced pluripotent stem cell-derived RPE,and some of them are undergoing clinical researches.Grafting manners in cell-based therapies are various including RPE sheet or RPE-choroid complex transplantation,RPE cell suspension injection,and RPE sheet transplantation with scaffolds.This review is limited to cell-based therapies for RPE that damaged first in the progress of AMD and focus on recent advances in cell sources,transplantation methods,preclinical and clinical trials,and the obstacles that must be overcome.

Objective To improve the diagnosis and therapy of uroseptic shock by percutaneous nephrolithotomy. Methods Five cases clinical data of uroseptic shock by percutaneous nephrolithotomy were analyzed retrospectively. Results Incidence rate of uroseptic shock by percutaneous nephrolithotomy was 0.5 % (5/1120), total deaths rate was 0.2 % (2/1120), death rate of uroseptic shock was 40%(2/5). Conclusions Uroseptic shock by percutaneous nephrolithotomy is extremely dangerous complication and primary death cause. Its characteristic such as: better masking, urgent onset,quick progress and high death rate. It is benefit to decrease incidence rate and mortality by prehension characteristic,control risk factor, early diagnosis and therapy.