1.Study on the mechanism of themolecular biology of hepatocyte growth factor and connective tissue factor in proliferative vitreoretinopathy
Chinese Journal of Primary Medicine and Pharmacy 2011;18(13):1733-1734,后插3
Objective To determine the role of CTGF in PVR, we successfully down - regulated the expression of CTGF by RNA interference( RNAi) technology in ARPE-19 cell line. Methods Design and synthesis three CTGF siRNA, screen effective targeted points in Western blot, combine Oligo DNA of effective targeted point, anneal to form double chain DNA, connect with linearing pGC-LV-GFP carrier, sequence DNA in PCR screening positive clones. Results The shRNA-expression vector of CTGF was constructed successfully. And RT-PCR and Western Blot results showed that CTGF expression was significantly inhibited by siRNA transfectants in ARPE-19 cells at mRNA and protein levels. Conclusion We successfully construct shRNA-expression vector of CTGF which paves a way for CTGF-targeted gene therapy of proliferative vitreoretinopathy.
2.Research progress in retinal vascular changes for cardiovascular disease assessment and prediction
Hongzhe ZHANG ; Weihong YU ; Youxin CHEN
Journal of Chinese Physician 2021;23(5):641-644
In recent years, the prevalence of cardiovascular disease (CVD) in China has been increasing year by year, and it has become a major public health concern. The prevention and control of chronic CVD is inseparable from effective early screening. However, due to the imbalance in the level of economic and social development in various parts of China, the universality and accessibility of screening in some areas still need to be improved. The fundus retinal blood vessel is the only vessel in the whole body that can be directly observed. Studies have found that its morphological and functional changes are related to the occurrence and development of many CVD, and it has the potential to become a tool for assessment and prediction. This review intends to introduce the research progress of retinal blood vessels in the evaluation and prediction of cardiovascular disease in recent years.
3.Clinical analysis of the synthesized treatment for elderly patients with serious burning
Delin HU ; Linsen FANG ; Youxin YU ; Changrong WANG ; Yongjie WANG
Chinese Journal of Geriatrics 2001;0(05):-
Objective To summarize the experience of early management in elderly patients with serious burning in order to raise cure rate. Methods Elderly patients (63 cases) with serious burning (burned area exceeding 30% or Ⅲ? area exceeding 10% ) admitted from 1990 to 2002 were retrospectively studied. Patients were divided into two groups according to the admission date, before and after January 1, 1996 since new management was extensively applied after 1996. These measures consisted of rapid and adequate fluid resuscitation, early enteral feeding and autograft after eschar excision. Results The incidence of sepsis and MODS was 38.7% and 19.4% respectively after 1996. It was evidently lower than the group before 1996(65.6% vs 43.8%, P
5.Application of artificial intelligence reading label system in diabetic retinopathy grading training of junior ophthalmologists and medical students
Ruoan HAN ; Weihong YU ; Huan CHEN ; Mingyue LUO ; Youxin CHEN
Journal of Chinese Physician 2021;23(5):650-653
Objective:To evaluate the efficiency of using artificial intelligence reading label system in diabetic retinopathy (DR) grading training among junior ophthalmologists and medical students.Methods:520 diabetic fundus images were randomly divided into 8 groups with 65 images in each group. 13 junior ophthalmologists and medical students were selected as the research objects. Each of them read 8 groups of pictures and evaluated the DR grading of each fundus image. The sensitivity, specificity and diagnostic test consistency (Q-kappa value) of grading results were analyzed with the DR grading given by 3 senior ophthalmologists as the gold standard. The average Q-kappa values of 13 subjects were compared between the first four times and the last four times.Results:Through 8 round reading, the average Q-kappa was elevated from 0.67 to 0.81. Average Q-kappa of round 1 to 4 was 0.77, and average Q-kappa of round 5 to 8 was 0.81. The participants were divided into two groups. Participants in group 1 were junior ophthalmologists and participants in group 2 were medical students. Average Q-kappa of group 1 was elevated from 0.71 to 0.76. Average Q-kappa of group 2 was elevated from 0.63 to 0.84.Conclusions:The artificial intelligence reading label system was a useful tool in training junior ophthalmologists and medical students in doing diabetic retinopathy grading.
6.Use of diagnostic vitrectomy in eyes with uveitis of unknown etiology
Haiyan LIN ; Rongping DAI ; Yannan LIN ; Fangtian DONG ; Meifen ZHANG ; Youxin CHEN ; Hanyi MIN ; Junjie YE ; Weihong YU ; Xiao ZHANG
Chinese Journal of Ocular Fundus Diseases 2017;33(4):404-406
Objective To analyze the results of diagnostic pars plana vitrectomy (PPV) in patients with uveitis of unknown cause.Methods This is a retrospective case series study.Sixty-five patients (67 eyes) with uveitis of unknown cause were enrolled in this study.There were 31 males (32 eyes) and 34 females (35 eyes).The ages were from 6 to 84 years,with the mean age of (55.00± 18.56) years.All eyes were received PPV.Examination of vitreous samples consisted of microbial stains and culture,microbial DNA and antibody detection,cytokine measurement,cytology,flow cytometry and gene rearrangement detection.Results Vitreous analysis was positive in 40 of 67 eyes (59.7%).Positive results indicated bacterial endophthalmitis in 20 of 40 eyes (50.0%),lymphoma in 11 eyes (27.5%),viral IgM and IgG increased significantly in 3 eyes (7.5%),fungal endophthalmitis in 3 eyes (7.5%),IgG of toxocara increased significantly in 2 eyes (5.0%),IgG of toxoplasma Gondii increased significantly in 1 eye (2.5%).Conclusion The diagnostic yield of vitreous samples in uveitis eyes of unknown cause is 59.7%.
7.Activity identification of anti-caspase-3 mRNA hammerhead ribozyme in both cell-free condition and BRL-3A cells
Renhuan XU ; Jing LIU ; Xiaqiu ZHOU ; Qing XIE ; Hong YU ; Youxin JIN ; Dan LIAO
Chinese Medical Journal 2001;114(6):606-611
Objective To study the transcription effects and cleavage activities of rat caspase-3 specific hammerhead ribozyme (Rz107) in both cell-free conditions and BRL-3A cells. Methods Rat caspase-3 gene fragment was cloned into the pGEM-T EASY vector under the T7 promoter control. The 32 P-labeled caspase-3 transcript was the target-RNA. Rz107 genes designed against caspase-3 mRNA were cloned into vector p1.5 between 5'-cis-Rz and 3'-cis-Rz. 32 P-labeled ribozyme transcripts were incubated with target-RNAs at different conditions and autoradiographed after denaturing gel-electrophoresis. Rz107 was electroporated into BRL-3A cells and the Rz107 expression was analyzed by RT-PCR. Results In cell-free conditions, Rz107 was active at 37℃. The optimal temperature was 50℃. The Km and kcat were 14.13?nmol/L and 2.31*min-1 respectively. Intracellular cleavage efficiency of Rz107 was 37%, as analyzed by RT-PCR. This indicated that the design of Rz107 was correct, and Rz107 had the activity of common enzymes. Conclusions Rz107 in cell-free conditions possessed perfect specific catalytic cleavage activity, and it can also cleave the target RNA successfully in cells. The results illustrate the feasibility of ribozyme therapy as a potential alternative approach for treating liver disease caused by apoptosis.
8.METTL3-mediated m6A methylation regulates lipopolysaccharide-induced endothelial cell permeability changes
Jianfeng WANG ; Huilin YU ; Youxin YU ; Junhui SONG ; Chengying MENG ; Wei JIANG ; Delin HU
Acta Universitatis Medicinalis Anhui 2024;59(6):1023-1028
Objective To explore the molecular mechanism of N6-methyladenosine(m6A)methylation mediated by methyltransferase 3(METTL3)in regulating lipopolysaccharide(LPS)-induced endothelial cell permeability changes.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro.HUVECs were treated with LPS 50,125,250,500,1 000,2 000 ng/ml for 24 h.METTL3 mRNA expression was detected by Real-time PCR.After HUVECs were intervened with 500 ng/ml for 24 h,the methylation level of m6A was detec-ted,and cell permeability was measured by cell permeability test.Real-time PCR and Western blot were used to detect mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-caherin).METTL3 overexpressed stable cell lines were constructed to measure the changes of m6A methylation level and per-meability of endothelial cells during METTL3 overexpression.Results Compared to the control group,LPS inhibi-ted the expression of HUVECs METTL3 mRNA,decreased the methylation of m6A,increased the cell permeabili-ty,and decreased the mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-Caherin).When METTL3 was overexpressed,the m6A methylation levels of endothelial cells were enhanced,and the increase of endothelial cell permeability induced by LPS was reversed.Conclusion METTL3-mediated m6A methylation can improve the permeability of endothelial cells induced by sepsis.
9.Validation and application of an artificial intelligence robot assisted diagnosis system for diabetic retinopathy
Shaohui GAO ; Xuemin JIN ; Zhaoxia ZHAO ; Weihong YU ; Youxin CHEN ; Yuhui SUN ; Dayong DING
Chinese Journal of Experimental Ophthalmology 2019;37(8):669-673
Objective To evaluate the performance of an artificial intelligence ( AI ) assisted diagnosis system for diabetic retinopathy ( DR) based on deep learning theory. Methods Diagnostic performance of a robot assisted diagnosis system called SongYue for DR was trained by using 25297 retinal images tagged by fundus doctors from multiple hospitals in China. Four types of DR detection model consisting of abnormal DR,referable DR,severe non-proliferative and proliferative DR as well as proliferative DR according to fundus leisions identification were established. The ability of the system to distinguish DR was determined by using receiver operator characteristic (ROC) analysis,sensitivity and specificity of the system. Results SongYue system achieved an area under the ROC curve ( AUC) of 0. 920 for successfully distinguishing normal images from those DR with a sensitivity of 96. 0%at a specificity of 87. 9%. The AUC of SongYue for referable DR was 0. 925,sensitivity was 90. 4%,and specificity was 95. 2%. For severe non-proliferative and proliferative DR,AUC was 0. 845,sensitivity was 72. 7%,and specificity was 96. 2%. For proliferative DR, AUC was 0. 855, sensitivity was 73. 5%, and specificity was 97. 3%. Conclusions SongYue robot assisted diagnosis system has high AUC,sensitivity and specificity for identifying DR, showing good clinical applicable benefits.
10.Microvascular changes of macular edema secondary to branch retinal vein occlusion treated with intravitreal conbercept injection
Zhikun YANG ; Weihong YU ; Youxin CHEN
Chinese Journal of Ocular Fundus Diseases 2021;37(9):675-680
Objective:To observe the changes of macular microvascular structure in eyes with macular edema secondary to branch retinal vein occlusion (BRVO-ME) after intravitreal injection of conbercept and analyze its relationship with visual function and central retinal thickness (CRT).Methods:A prospective clinical study. From July 2018 to June 2019, 21 eyes of 21 patients with unilateral temporal BRVO-ME diagnosed in the Department of Ophthalmology of Peking Union Medical College Hospital were included in the study. Among them, there were 14 eyes of 14 males and 7 eyes of 7 females; the average age was 58.0±8.3 years. There were 13 eyes and 8 eyes with occlusion of the superior temporal and inferior temporal branches of the retinal vein, respectively. The affected area was defined as the side of the venous obstruction. All the affected eyes underwent best-corrected visual acuity (BCVA) and optical coherence tomography angiography (OCTA) examination. The BCVA was tested using the international standard logarithmic visual acuity chart, which was converted into the logarithmic minimum angle of resolution (logMAR) visual acuity during statistical analysis. All the eyes were treated with intravitreal injection of conbercept once a month for 3 months, and then treated as needed. A 3 mm × 3 mm scan centered on fovea was obtained and the vascular density of superficial capillary plexus (SCP) and deep capillary plexus (DCP), fovea avascular zone (FAZ) area, perimeter of FAZ (PERIM), acircularity index (AI), foveal vascular density in a 300 μm wide region around FAZ (FD-300) and central retinal thickness (CRT) were measured. The follow-up time after treatment was 6 months. The vascular density and FAZ parameters were compared before and after treatment by paired t test. The correlations of BCVA, CRT and vascular density, FAZ area and the other parameters at 6 months after treatment were analyzed by linear regression analysis. Results:Before treatment, the logMAR BCVA of the eyes was 0.506±0.159, and the CRT was 375.4±81.3 μm; 6 months after treatment, the logMAR BCVA of the eyes was 0.294±0.097, and the CRT was 266.3±46.7 μm. There was a statistically significant difference of logMAR BCVA and CRT between the eyes before and after treatment ( t=8.503, 9.843; P<0.05). There was no statistically significant difference in the overall vascular density of SCP and DCP before and 6 months after treatment ( t=-0.091, -0.320; P>0.05). The foveal vascular density decreased, and the difference was statistically significant ( t=8.801, 3.936; P<0.05). The vascular density of DCP of the affected area increased, and the difference was statistically significant ( t=-2.198, P<0.05). Compared with those before treatment, the FAZ area and PERIM of the affected eyes had an increasing trend, while AI and FD-300 had a decreasing trend, and the differences were statistically significant ( t=-18.071, -12.835, 2.555, 8.610; P<0.05). The linear regression analysis showed that BCVA and FAZ area 6 months after treatment have significant correlation ( t=2.532, P=0.024). Conclusion:CRT decreased and BCVA increased after intravitreal injection of conbercept in BRVO-ME eyes. After treatment, the foveal vascular density of SCP and DCP decreased while the vascular density of DCP of the affected area increased. The FAZ increased and the PERIM and AI decreased during follow-up. The BCVA was significantly correlated with the FAZ area 6 months after treatment.