1.Blue-light-induced apoptosis of cultured human retinal pigment epithelial cells in vitro
Shanjun CAI ; Mi YAN ; Junjun ZHANG
Chinese Journal of Ocular Fundus Diseases 2003;0(06):-
Objective To observe the effect of blue light on apoptosis of cultured human retinal pigment epithelial (RPE) cells in vitro. Methods Human RPE cells were exposed to blue light, and the cells were divided into 3 groups: group A, with various intensity of illumination; group B: with same intensity but different time of illumination; group C: with same intensity and time of illumination but different finish time of the culture. The apoptosis of RPE cells was observed by TdT-dUTP terminal nick-end labeling (TUNEL) and annexin V-fluoresein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry, and transmission electron microscopy. Results The positive cells stained by TUNEL shrinked and turned round, whose nuclei concentrated and congregated like the crescent or hat. Cracked nuclei and membrane bleb were found. Swollen mitochondrial, disappeared inner limiting membrane of mitochondria, and dilation of the rough endoplasmic reticulum with metabolite were observed by transmission electronmicroscopy. In group A, mild damage of RPE cells was found when the threshold value of the intensity of illumination was less than (500?100)lx, and the apoptosis and necrosis of RPE cells aggravated as the intensity of illumination increased; in group B, as the time of illumination extended, the number of apoptotic RPE cells didn′t increase while the necrosis increased; in group C, 6 and 12 hours after illumination, apoptosis of cells was the main injury, while apoptosis with necrosis was found and necrotic cells increased as the time of illumination was prolonged. Conclusions Illumination with blue light may cause damages of human RPE cells in vitro, with the modalities of apoptosis, apoptotic necrosis and necrosis. The extent of injury is dependent on intensity and duration of the illumination.
2.The clinical features of polypoidal choroidal vasculopathy
Shanjun CAI ; Jian TANG ; Yingjie ZHAO
Chinese Journal of Ocular Fundus Diseases 2003;0(05):-
Objective To investigate the clinical features of polypoidal choroidal vasculopathy (PCV) in Chinese patients. Methods The clinical data of 71 continuous patients (142 eyes) who were diagnosed with PCV by fundus photography, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA) and optical coherence tomography (OCT) were analyzed retrospectively. Results Eleven patients (11 eyes) of 71 patients (142 eyes) were diagnosed with PCV to make up 15.49% and 7 75% of the numbers of patients and affected eyes respectivery. The patients included 7 males (63 6%) and 4 females (36 4%). The predominant location for these lesions was the macular region in 10 eyes (90 9%). Fundus examination demonstrated the reddish-orange nodular elevations in 6 eyes. ICGA revealed umbrellalike or twiglike branching vascular networks and polypoidal dilations at the vascular terminals of the branches in all patients, and feeder vassels within choroidal vascular networks were found in 8 eyes. OCT images of retinochoroidal structures showed prominent anterior protrusion of the orange subretinal mass corresponding to the polypoidal structure in the indocyanine green angiogram. An apparent discontinuity was observed in the highly reflective layer which delineates the polypoidal structure. Conclusions PCV mainly affects the male over 50 years and the eyes involved were mostly unilateral. Most of polypoidal vascular lesions are present in the macular area.
3.Inhibition of apoptosis induced by galactose by c-myc antisense oligodeoxynucleotide in lens epithelial cells
Fang, FAN ; Honghua, LIN ; Zhenglong GE ; Huaqing, LIU ; Xiaoping ZENG ; Shanjun, CAI ; Haixiang LI
Chinese Ophthalmic Research 2010;28(3):203-206
Background Various studies demonstrated that the apoptosis of lens epithelial cells(LECs) is associated with the overexpression of the c-myc gene in LECs induced by galactose.Inhibiting the abnormal expression of the c-myc gene in LECs is an effective approach to mitigate the pathogenesis and development of cataract.Objective The goal of this study is to investigate the inhibitory effects of c-myc antisense oligodeoxynucleotide(c-myc ASODN) on the apoptosis of LECs in the eye with galactose-induced cataract.Methods Galactose-induced cataract models were established by the retrobulbar injection of 0.2 mL of 20% galactose once per day.Lipo-antisense oligodeoxynucleotide(Lipo-ASODN,0.2 mL) was retrobulbarly injected 4 hours after the injection of galactose at one-day intervals.The animals were sacrificed and lenses were obtained to evaluate the apoptosis of LECs and the effect of c-myc ASODN on LECs apoptosis induced by galactose was examined by TUNEL assay after 7,14 and 24 days.The ultrastructural changes of LECs were examined under the transmission electron microscopy(TEM).Results A significant difference in the apoptotic rate of LECs was found among the 7 day,14 day and 24 day groups(F_(7 days)=3 418.495,P<0.01;F_(14 days)=1137.555,P<0.01;F_(24 days)=2198.871,P<0.01).The apoptotic rate of LECs in the galactose group was markedly higher than that in the normal saline solution group 7 days,14 days and 24 days after the experiment(P<0.01).The apoptotic rate of LECs in the galactose+lipo+ ASODN group significantly declined in comparison to the galactose group after 7 days,14 days and 24 days(P<0.05).TUNEL assay showed the condensation,breakage and irregularity of the nuclei of apoptotic cells in the galactose group.The destruction of the ultrastructure of the cells and organelles were observed under the transmission electron microscope.Conclusion Galactose induces apoptosis of LECs in cataractogenesis.C-myc ASODN inhibits apoptosis of LECs induced by galactose.
4.Mitochondrial pathway of retinal pigment epithelial cell apoptosis induced by blue light in vitro
Hong, LI ; Jianping, LYU ; Shanjun, CAI ; Gang, SU ; Zhipeng, WU ; Xin, GONG
Chinese Journal of Experimental Ophthalmology 2015;33(1):16-20
Background Studies determined that blue light exposure causes apoptosis of human retinal pigment epithelial (RPE) cells,but its mechanism is still below understood.Objective The aim of this study was to investigate whether or how mitochondrial apoptotic pathway is involved in blue-light induced apoptosis of human RPE cells in vitro.Methods Human RPE cells were isolated from fresh donor eyes and primarily cultured and passaged.The cells were identified with keratin antibody by immunochemistry.Then the cells were the non-light exposed group,simple light-exposed group,light-exposed+nifedipine group,light-exposed+calphostin C group and the light-exposed+phorbol myristate acetate (PMA) group.Human RPE cells in light-exposed group were consequently cultured for 24 hours following the exposure of (2 000±500)lx blue-light for 6 hours,and then the expression levels of bax,bcl-2,bcl-xl in the cells were detected by Western blot to evaluate the effect of blue light on the apoptosis.The cells in the light-exposed+nifedipine group,light-exposed+calphostin C group and the light-exposed+PMA group were treated with the corresponding drugs 1 hour prior to light irradiation and sequently received 6-hour light irradiation and 48-hour culture.The expression of caspase-9 protein in the cells were assayed with Western blot to assess the influence of Ca2+ channel and protein kinase C (PKC) pathway on mitochondria of RPE cells.Results Cultured cells grew well with visible pigment in cytoplasm.The cells showed the positive response for keratin and presented a cobblestone-like appearance.The expression bands of bax,bcl-2 and bcl-xl proteins were clearly visible at the molecular weight of 23 000,26 000 and 30 000 in both non-light exposed group and the simple light-exposed group,and the absorbance values of the cells to bax were elevated,while the absorbance values to bcl-2 and bcl-xl were declined in the simple light-exposed group compared with the non-light exposed group (t =-4.409,P =0.012 ;t =7.575,P =0.002 ; t =6.068,P =0.004).Compared with the non-light exposed group,the absorbance values of caspase-9 were significantly raised in the simple light-exposed group,light-exposed+calphostin C group and the lightexposed+PMA group (P=0.005,0.002,0.000),but no significant difference between the non-light exposed group and light-exposed+nifedipine group (P=0.191).Compared with the simple light-exposed group,the expression level was considerably higher in the light-exposed + PMA group (P =0.005) ; while that in the light-exposed + nifedipine group or light-exposed+calphostin C group was not significantly different (P=0.057,0.643).Conclusions Blue light exposure induces apoptosis of RPE cells by up-regulating the expressions of bax and caspase-9 proteins and down-regulating the expressions of bcl-2 and bcl-xl.The mitochondrial apoptosis pathway and PKC pathway participate in blue-light induced apoptosis of human RPE cells in vitro.
5.Lymph node metastasis and prognostic analysis of early gastric cancer patients
Yingchao WU ; Miao XIE ; Yunlong CAI ; Tao WU ; Shanjun HUANG ; Xin WANG
Chinese Journal of General Surgery 2019;34(7):561-564
Objective To investigate the correlation between clinicopathological features and lymph node metastasis in early gastric cancer (EGC) and evaluate the influence of lymph node metastasis on the overall survival.Methods The clinicopathological data of 178 EGC patients undergoing radical gastrectomy with lymphadenectomy in Peking University First Hospital between Jan 2006 and Dec 2015 were retrospectively reviewed.The impact of lymph node metastasis on the overall survival was analyzed.Results Lymph node metastasis was detected in 19 (10.7%) of 178 patients.Univariate analysis showed a positive relationship between tumor size (x2 =4.804,P =0.028),depth of invasion (x2 =8.176,P =0.003),histological type (x2 =4.333,P =0.037),vascular tumor thrombus (x2 =9.992,P =0.002) and lymph node metastasis in EGC.Multivariate analysis revealed that depth of invasion (Wald =4.954,P =0.026)and intra-vascular tumor thrombus (Wald =3.966,P =0.046) were independent relative factors of lymph node metastasis in EGC patients.The 5-year survival rates of EGC patients with lymph node metastasis were 78.9%,much lower than that without lymph node metastasis (94.3%,x2 =8.310,P=0.004).Conclusions Lymph node metastasis in EGC is mainly correlated with depth of invasion and intra-vascular tumor thrombus.The prognosis of patients with lymph node metastasis is significantly poorer than those without lymph node metastasis.
6.Research status and progress of endoplasmic reticulum stress in scleral remodeling
Zhao LIU ; Bing XIE ; Shanjun CAI
Chinese Journal of Ocular Fundus Diseases 2023;39(10):873-878
The occurrence and development of myopia is closely related to scleral remodeling. Therefore, in order to effectively prevent and cure myopia, it is very important to clarify the mechanism of scleral remodeling. In recent years, Chinese scholars have found that endoplasmic reticulum stress can regulate the expression of apoptotic proteins through the inositol demand protein-1/X box binding protein-1 pathway in the unfolded protein response, thus it is involved in regulating the state of scleral fibroblasts under hypoxia and regulating the occurrence and development of scleral remodeling. At the same time, some studies have found that inhibiting and knocking out protein kinase RNA-like endoplasmic reticulum kinase and activated transcription factor 6 in endoplasmic reticulum stress can effectively inhibit the growth of ocular axis. This proves that endoplasmic reticulum stress plays an important role in the occurrence and development of scleral remodeling. However, the comprehensive analysis of endoplasmic reticulum stress and scleral remodeling has not been reported at home and abroad. In-depth analysis of the relationship between endoplasmic reticulum and scleral remodeling is of great significance for the follow-up analysis and study of the mechanism of scleral remodeling.
7.Elevated blood pressure increases risk of proliferative diabetic retinopathy: a Mendelian randomization study
Gang SU ; Zhao LIU ; Shanjun CAI
Chinese Journal of Ocular Fundus Diseases 2024;40(2):142-147
Objective:Mendelian randomization (MR) was used to analyze the potential relationship between blood pressure and proliferative diabetic retinopathy (PDR).Methods:Two-sample MR analysis was performed using summary statistics from genome-wide association studies. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were selected as the exposure, PDR as the outcome. The instrumental variable of SBP and DBP came from the publicly available data of the the UK Medical Research Council Comprehensive Epidemiology Unit and Neale Laboratory; the outcome data (8 681 cases in the case group, 204 208 cases in the control group, European population) are from the FinnGen database. Inverse variance weighting (IVW) and weighted median (WM) were used to analyze the potential relationships between SBP, DBP and PDR.Results:MR analysis showed that IVW [SBP: odds ratio ( OR)=1.36, 95% confidence interval ( CI) 1.17-1.57, P= 4.22E-05; DBP: OR=1.29, 95% CI 1.11-1.51, P=8.6E-04], WM (SBP: OR=1.33, 95% CI 1.07-1.66, P=0.009; DBP: OR=1.28, 95% CI=1.03-1.59, P=0.002). The results showed that elevated SBP and DBP increased the risk of PDR. Conclusion:Blood pressure (SBP, DBP) change is positively correlated with the risk of PDR.
8.Long-term follow-up observation after vitrectomy in a family with vitreous amyloidosis due to transthyretin gene Gly83Arg mutation
Hong LI ; Xingwang CHEN ; Gang SU ; Huixuan REN ; Yue GOU ; Mo JIANG ; Xiaomei NIE ; Bin XIE ; Shanjun CAI
Chinese Journal of Ocular Fundus Diseases 2021;37(6):418-422
Objective:To investigate the causes of secondary glaucoma after vitrectomy for familial vitreous amyloidosis associated with transthyretin (TTR) gene Gly83Arg mutation.Methods:A retrospective case study. From January 2008 to January 2020, 13 cases (23 eyes) with hereditary vitreous amyloidosis and treated by vitrectomy in the Affiliated Hospital of Zunyi Medical University were collected. Among them, there were 7 males with 12 eyes and 6 females with 11 eyes. The average age was 43.0±4.8 years. All the affected eyes underwent standard three-channel vitrectomy through the flat part of the ciliary body. According to whether complete vitreous detachment (PVD) was formed during the operation, it was divided into complete PVD group and incomplete PVD group; according to the occurrence time of secondary glaucoma and vitreous amyloidosis after surgery, it was divided into 1-12 months group and 13-36 months group, >37 months group. The average follow-up time after surgery was 36.7±6.0 months. The incidence of secondary glaucoma and the recurrence rate of vitreous amyloidosis between groups were compared by χ2 test; the correlation between recurrence of vitreous amyloidosis and secondary glaucoma after surgery was analyzed by Spearman rank correlation analysis. Results:Among the 23 eyes, there were 8 eyes in the complete PVD group and 15 eyes in the incomplete PVD group, respectively. Vitreous amyloidosis recurred in 15 eyes (65.22%, 15/23) after surgery. There were 14 (93.30%, 14/15) and 1 (6.70%, 1/15) eyes in the incomplete PVD group and the complete PVD group, respectively; the comparison of the recurrence rate of vitreous amyloidosis between the two groups was statistically significant ( χ2=11.676, P<0.01). 1-12 months group, 13-36 months group, >37 months group included 1 (4.35%, 1/23), 12 (52.17%, 12/23), 2 (8.70%, 2/23) Only eye. The recurrence rate in the 13-36 months group was significantly higher than that in the 1-12 months group and >37 month group. Secondary glaucoma occurred in 11 eyes (47.80%, 11/23) after surgery. 1-12 months group, 13-36 months group, above 37 months group were 1 (4.35%, 1/23), 8 (34.78%, 8/23), 2 (8.70%, 2/23) eyes. The incidence of secondary glaucoma in the 13-36 months group was higher than that in the 1-12 months group and >37 months group. Among 11 eyes with secondary glaucoma, 10 eyes had recurrence of vitreous amyloidosis after surgery, and 1 eye had no recurrence. The results of Spearman rank correlation analysis showed that there was a positive correlation between the recurrence of vitreous amyloidosis and the occurrence of secondary glaucoma ( rs=0.516, P=0.012). Conclusion:The incidence of secondary glaucoma after vitrectomy in a family with vitreous amyloidosis caused by the Gly83Arg mutation of TTR gene is higher, and its occurrence is significantly positively correlated with the recurrence of vitreous amyloidosis.
9.Analysis of refractive prediction errors after flanged intrascleral intraocular lens implantation
Recent Advances in Ophthalmology 2024;44(4):333-336
Flanged intrascleral intraocular lens implantation has become one of the mainstream treatment solutions for aphakia in clinical practice due to its ability to avoid dislocation of the intraocular lens,or subluxation caused by suture degradation or breakage,as well as the ability to eliminate complications such as inflammation or infection caused by su-tures,and the significant improvement in postoperative vision for patients.However,there has been no systematic analysis of the factors that may lead to postoperative refractive prediction errors in China.This article analyzes the possible prob-lems of flanged intrascleral intraocular lens implantation and the reasons for postoperative refractive prediction errors,in order to provide reference for clinical work.
10.Causality between body mass index and central retinal artery occlusion:a Mendelian randomized study
Zhao LIU ; Li ZHOU ; Qian XIE ; Shanjun CAI
Recent Advances in Ophthalmology 2024;44(3):193-196
Objective To analyze the causal relationship between obesity and central retinal artery occlusion(CRAO)based on body mass index(BMI),which is commonly used to measure obesity,and Mendelian randomization(MR)method.Methods Two-sample MR analysis was performed using summary statistics from genome-wide associa-tion studies.Exposed genetic tools were divided into the men group(n=60 586,SNP=2 736 876,European men)and women group(n=171 977,SNP=2 494 613,European women);CRAO was selected as the outcome.The instrumental variable for BMI came from the Integrative Epidemiology Unit(IEU)publicly available data;the outcome CRAO data came from the FinnGen database.Inverse variance weighting(IVW)and weighted median(WM)were used to analyze the poten-tial relationship between BMI and CRAO.Results IVW showed that the increased BMI level significantly increased the risk of CRAO in men[OR=4.57,95%CI:1.32-15.82,P=0.016]and the risk of CRAO in women[OR=3.48,95%CI:1.40-8.63,P=0.007].Meanwhile,the WM and MR-Egger analysis results supported the above conclusions.In addition,there was no heterogeneity or pleiotropy in the results of this study.Conclusion The increase in BMI is positively corre-lated with the occurrence of CRAO.This study provides an effective CRAO prevention strategy for asymptomatic patients with elevated BMI levels.