Background Choroidal neovascularization (CNV) is one of the causes of blindness in multiple eye diseases.Researches showed that complement system participates in the pathogenesis of CNV.Objective This study was to construct the recombinant of complement factor B-small interference RNA (CFB-siRNA) expression vector and to observe its inhibitory effect on human umbilical vein endothelial cells (ECV-304).Methods CFB gene primers were designed based on human CFB gene,and an expression vector of CFB-siRNA was constructed by inserting CFB-siRNA into pRNAT-U6.1/Neo plasmid.Recombinant plasmids were confirmed by the digestion analysis of restriction endonuclease,and all inserted sequences were verified by DNA sequencing.The recombinant pRNAT-U6.1/CFB-siRNA plasmid and the blank plasmid were transfected into ECV-304 cells in the CFB-siRNA group and blank plasmid group by electroblot,respectively,and non-transfected cells served as the normal control group.The cells were observed under the fluorescence microscope 48 hours after transfection,and the transfective efficiency was calculated.The relative expression of CFB mRNA in the cells of different groups was detected by semi-quantitative reverse transcription PCR (RT-PCR).MTT was employed to calculated the growth inhibitory rates of the cells 24,48 and 72 hours after transfection.The percentages of the cells in different cell cycles were detected by flow cytometry.Results The sequence of the target vector was identical to the designed sequence.The green fluorescence protein (GFP) was seen in both the CFB-siRNA group and the blank plasmid group.The relative expression levels of CFB mRNA were 0.07 ±0.04,0.14 ±0.02 and 0.14 ±0.03 in the CFB-siRNA group,the blank plasmid group and the normal control group,respectively,a significant difference was obtained among the three groups (F=233.05,P =0.00);the expression level of CFB mRNA in the CFB-siRNA group was significantly declined in comparison with the blank plasmid group and the normal control group (both at P＜0.05).The growth inhibitory rates of the cells were (23.45 ±0.01) ％,(33.48 ±0.02) ％ and (45.49±0.01) ％ at 24,48 and 72 hours after transfection,respectively,a significant difference was obtained among the three groups (Fgroup =212.99,P =0.00);the growth inhibitory rates in CFB-siRNA group were significantly higher than that in the blank plasmid group and normal control group (all at P＜ 0.05).The percentages of G1 phase cells were (44.4 ±0.5) ％,(25.8 ±0.4) ％ and (27.9 ± 0.6) ％ in the CFB-siRNA group,the blank plasmid group and the normal control group respectively,a significant difference was obtained among the three groups (F=58.98,P=0.00).The percentages of G1 phase and G2 phase cells in the CFB-siRNA group were significantly higher than those in the blank plasmid group and the normal control group (all at P＜0.05).Conclusions Recombinant pRNAT-U6.1/CFB siRNA inhibits the proliferation of ECV-304 cells effectively by arresting the cells in G1 intermediate phase of the growth cycle.

Objective To investigate the risk factors associated with neovascular glaucoma (NVG) after pars plana vitrectomy (PPV) in eyes with proliferative diabetic retinopathy (PDR).Methods Retrospective study.One hundred and thirty-seven patients (137 eyes) with PDR who underwent PPV were recruited.There were 85 males and 52 females.The average age was (60.1 ± 8.8) years old.The duration of diabetes was (10.2 ± 3.6) years.There were 49 patients with ipsilateral carotid artery stenosis.Fifty-three eyes underwent intravitreal ranibizumab or conbercept injection before PPV.All eyes were treated with 23G standard three-port PPV.The average follow-up time after PPV was 11.5 months.Fundus fluorescein angiography (FFA) was conducted in postoperative 4-6 weeks to observe non-perfused retinal areas.Risk factors,such as ipsilateral carotid artery stenosis,the presence of non-perfusion in retina after PPV and the application of anti-vascular endothelial growth factor (VEGF) drugs before PPV,were identified by logistic regression.Results Twenty of 137 patients (14.6％) developed postoperative NVG after PPV.Ipsilateral carotid artery stenosis [odds ratio (OR) =5.048,95％ confidence interval (CI) 2.057-12.389,P=0.000] and the presence of non-perfusion in retina after PPV (OR=4.274,95％CI 1.426-12.809,P=0.009) were significant risk factors for postoperative NVG,while the application of anti-VEGF drugs was not (OR=1.426,95％CI 0.463-4.395,P=0.536).But the time from PPV to the onset of NVG varies significantly between the two groups of injection of anti-VEGF drugs or not (t=-4.370,P=0.000).Conclusions Risk factors associated with NVG after PPV in eyes with PDR included ipsilateral carotid artery stenosis and the presence of non-perfusion in retina after PPV.The application of anti-VEGF drugs before PPV can delay the onset of NVG in PDR eyes after vitrectomy.

Objective To observe the efficacy ofvitrectomy combined with internal limiting membrane (ILM) peeling and scleral shortening for myopic foveoschisis (MF).Methods Prospective and non-randomized concurrent control study.A total of 35 MF patients (35 eyes) were enrolled in this study.The patients were divided into 2 groups according to surgery,including group A (18 eyes) and group B (17 eyes),all received vitrectomy combined with ILM peeling,but group A also received scleral shortening.The best corrective visual acuity (BCVA) examination using the Snellen vision chart was converted to the minimum resolution logarithm (logMAR).Ocular axis length (AL) was measured by Zeiss IOL-Master or A-scan ultrasound (Quantel Medical,France).The maximal value of retinal foveoschisis (MxFT) was measured by frequency-domain optical coherence tomography (Heidelberg,Germany).Multifocal electroretinogram (mfERG) responses were obtained with the RETIscan system (Roland Consult,Gemany).There was no statistically significant difference between the two groups (P＞0.05) in age (t=0.460),AL (t=1.520),diopter (t=0.020),logMAR BCVA (t=-2.280),MxFT (Z=-4.179) and b-wave ERG amplitude (Z=-0.198).The changes of BCVA,AL,MxFT and b wave amplitude were followed-up for 3-12 months.Results At the last follow-up,the height of MF was decreased in 18 eyes of group A,and MF was completely disappeared in 4 eyes.The logMAR BCVA (t=7.272,5.951),MxFT (Z=-3.724,-3.622) and b-wave ERG amplitude (Z=-3.223,-3.243) in both groups A and B were statistically improved (P=0.000,0.000,0.000,0.000,0.001,0.001) compared to pre-operational results.There was significant difference of logMAR BCVA (t=-2.280) and MxFT (Z=-4.179) between the two groups (P=0.029,0.000).But there was no significant difference in the amplitude of b-wave (Z=-0.198,P=0.843).The AL in group A was shortened after surgery,the difference was statistically significant (t=10.017,P=0.000).During the follow-up,there was no ocular hemorrhage,endophthalmitis and other complications.Conclusion PPV combined with ILM peeling and scleral shortening can shorten AL significantly for MF patients,and gain relative normal anatomical structure of the fovea,thus improve the vision.

Objective To investigate the effects of liposome mediated intraocular gene transfection of endostatin on the inhibition of the development of choroidal neovascularization (CNV) in a rat model. Methods Experimental CNV model in Brown Norway rats was induced by laser photocagulation. The recombinant eukaryotic expression plasmid pSecTagA hEndostatin or control plasmid pSecTagA and liposome complexes were injected into the subretinal space of the model rats. In situ hybridization and immunohistochemical observation confirmed the presence of endostatin mRNA and protein expression two weeks after injection. Intraocular and serum levels of endostatin were measured by enzyme linked immunosorbent assay. The intensity of fluorescein leakage from the photocoagulated lesions was studied at 13 d after photocoagulation. The area of CNV was measured using high molecular weight FITC dextran (MW2?10 6) for high resolution angiography in RPE choroid sclera flat mounts. In addition, sections of CNV lesions were studied by light microscopy and endoglin (CD105) immunohistochemical evaluation. Results The retina, RPE, choroidal were infected by subretinal delivery of the pSecTag hEndostatin and expressed the endostatin. Two weeks after intraocular injection, the level of endostatin in the whole eye homogenates were (50 14?3 43) ng/eye and (31 5?2 21) ng/eye, respectively. Fluorescein leakage from the CNV lesions decreased significantly as compared with that in the control groups. The average area of CNV at the sites of the Bruch's membrane rupture showed significant difference in eyes injected with endostatin as compared with that in the control eyes. Endothelial cells demonstrated strong immunoreactivity of CD105 in CNV lesions in the control eyes. Conclusion Liposome mediated endostatin gene transfection can significantly inhibit the development of CNV.

Objective:To observe the changes of the structure and visual function of the retina in patients with or without the ectopic inner foveal layers (EIFL) and to explore the factors influencing the recovery of visual function in patients with idiopathic epimacular membrane (IMEM).Methods:A retrospective clinical study. From March 2015 to June 2019, 90 patients with MEM who were diagnosed by Ophthalmic Center of the Second Hospital of Hebei Medical University were enrolled in the study. All patients were examined by best corrected visual acuity (BCVA) and frequency domain optical coherence scan. BCVA was recorded by Snellen vision table, and it was converted into the minimum resolution angle logarithm (logMAR) vision. Among 90 eyes, IMEM grade 2-4 was 68 (75.6%, 68/90), 18 (20.0%, 18/90), 4 (4.4%, 4/90), respectively. According to this, the grade 2 was set as group A, and the grade 3 and grade 4 were combined to group B. There was no significant difference in age ( t=0.015), sex composition ratio of patients between two groups ( χ2=0.060) and the average of central macular thickness (CMT) ( F=2.277) ( P=0.904, 0.809, 0.141). The difference of average logMAR and BCVA was statistically significant ( F=35.913, P=0.000). All patients underwent 25G pars plana three channel vitrectomy with simultaneous removal of epiretinal membrane and internal limiting membrane. BCVA, CMT and improvement of IMEM grading were observed at 1, 3, 6 and 12 months after operation. BCVA, EIFL thickness and CMT were compared before and after operation by single factor repeated variance analysis; Fisher exact probability method was used to compare the changes of the anatomical structure of the eyes in the two groups at 12 months after operation. Results:1, 3, 6, 12 months after operation, the average eyes of logMAR BCVA in group A were 0.50±0.13, 0.38±0.12, 0.27±0.12, 0.19±0.10. The patients in group B were 0.66±0.14, 0.60±0.13, 0.54±0.14, 0.52±0.14. CMT in group A were 364.82±81.29, 281.65±72.45, 228.55±55.34, 182.84±56.13 μm. The patients in group B were 455.88±69.60, 440.18±68.65, 383.76±65.38, 371.39±66.60 μm. The difference was statistically significant in the two groups (BCVA: F=37.913, 11.479, 24.250, 39.013; P=0.000, 0.002, 0.000, 0.000. CMT: F=10.987, 39.610, 55.789, 79.987; P=0.002, 0.000, 0.000, 0.000). In group A, IMEM was improved to 57 eyes of grade 1 on 12 months after operation. Among the 18 eyes in group B, IMEM was improved to 1 and 3 eyes in level 1 and level 2, respectively, and no improvement was found in 4 eyes in grade 4. The difference was statistically significant ( P=0.000) in the improvement of the number of eyes in the two groups. Conclusions:The patients with IMEM without EIFL have better visual prognosis and reversible anatomical changes. EIFL is an important factor affecting the visual function and anatomical structure recovery after operation.

This study was aimed to search anti-platelet aggregation effectors from Gardenia jasminoides extract with the employment of platelet affinity extraction method coupled with HPLC, in order to provide pharmacological experi-mental evidences of the selected effectors to verify its feasibility. Under physiological conditions, washed rat platelets were added into G. jasminoides extract and then a mixture was gained. Consequently, some components from G. jas-minoides extract were combined to the platelets in the mixture while some were not owing to their special chemical structures and properties. Firstly, the uncombined components were washed off from the mixture. Secondly, the com-bined components in the leftover was washed down and collected, respectively, right after destroying the occupied platelets' structures. Thirdly, different collected eluents were analyzed, respectively, by HPLC established in the pre-vious work to search the effectors. Fourthly, pharmacological experiments were implemented for confirmation. The re-sults showed that dominant effective components from G. jasminoides extract acting on anti-platelet aggregation were identified as geniposide. Further evident was provided as well by pharmacological experiment that geniposide exhibit-ed significant inhibitory effect on anti-platelet aggregation in rats induced by ADP, rat tail collagen and thrombin(P< 0.01). It was concluded that the platelet affinity extraction-HPLC method proposed in this paper can be utilized to analyze the correlation of effectors from G. jasminoides extract and its pharmacological effects. Moreover, there are some correlations between screened chemical substances and their pharmacological effects.

Objective:To summarize the clinical manifestations and color Doppler ultrasonography of benign lymphoepithelial disease (BLL) in lacrimal gland, so as to improve the diagnostic accuracy of lacrimal BLL.Methods:Clinical manifestations and color Doppler flow imaging (CDFI) features of lacrimal BLL in 16 patients (2 males and 14 females, with 31 lesions) who visited the Second Hospital of Hebei Medical University from November 2014 to August 2019 were retrospectively analyzed.Results:The performance for lesions in patients with duration less than 3 months was scattered in irregular low echo, lesion blood flow signals within the rich (Adler Ⅱ level), pathologic examination results showed more lymphocytes were seen in the lacrimal matrix, and the myoepithelium of the lacrimal duct grew to form the epithelium-myocutaneous island. The ultrasonic feasures in patients with course of 3-6 months were in multiple categories such as circular low echo, "honeycomb" change, CDFI showed lesions with a lot of blood flow signals (Adler Ⅲ level), the pathological examination results indicated that there were a large number of lymphocytes and epithelial-musculocutaneous islands in the lacrimal matrix, and the lymphocytes were significantly increased compared with patients with the course of disease less than 3 months. The ultrasound results in patients with a course of more than 6 months showed lesions in multiple categories such as circular low echo, and large low echo, greater than 3 mm in diameter, CDFI showed lesions within the same large amounts of blood flow signals (Adler Ⅲ level), the pathological examination results were consistent with the course of 3 to 6 months. One patient had positive tuberculin test and 11 had higher IgG4 than normal.The number of lymphocytes increased with the prolongation of disease course.Conclusions:With the expert knowledge of color Doppler ultrasonographic characteristics of lacrimal gland BLL and with the serum IgG4 level helps to improve the accuracy of ultrasonic diagnosis.

Objective To observe the expression ofRapl,guanosine triphosphate-Rapl (GTP-Rapl),vascular endothelial growth factor (VEGF) and β-catenin in experimental choroidal neovascularization (CNV).Methods Forty-two brown Norwegian rats were randomly divided into a blank control group (7 rats) and a model group (35 rats).Both eyes were enrolled.The CNV model was established by holmium ion laser photocoagulation in the model group.At 3,7,14,21,and 28 days after photocoagulation,fluorescein fundus angiography (FFA) and choroidal vascular smear were performed to observe the degree of fluorescein leakage and CNV area in rats;Western blot and real-time quantitative polymerase chain reaction (RT-PCR) were used to detect the expression ofRap1,GTP-Rap1,VEGF,β-catenin and mRNA in CNV.Results The results of FFA examination showed that a large disc-shaped fluorescein leaked in the photo-condensation spot 14 days after photocoagulation.Laser confocal microscopy showed that compared with 7 days after photocoagulation,CNV area increased at 14,21,28 days after photocoagulation,and the difference were statistically significant (t=3.725,5.532,3.605;P＜0.05).Western blot showed that there was no significant difference in the relative expression of Rap1 protein in CNV at different time points after photocoagulation between the two groups (P=0.156).Compared with the blank control group,the relative expression of GTP-Rap1 protein was significantlydecreased,the relative expression of VEGF and β-catenin protein were significantly increased in the model group (P=0.000).The results of RT-PCR showed that there was no significant difference in the relative expression of Rap 1 mRNA at different time points after photocoagulation between the two groups (P=0.645),but there were significant difference in the relative expression of β-catenin mRNA (P=0.000).At 7,14,21 and 28 days after photocoagulation,there were significant difference in the relative expression of GTP-Rap 1 and VEGF mRNA between the two groups (P=0.000).Conclusions The expression of GTP-Rap1 in experimental CNV is significantly lower than that in normal rats.

Perifoveal exudative vascular anomalous complex (PEVAC) are class of diseases characterized by isolated aneurysmal lesions of the perifovea with hemorrhage and hard exudates, which are found in recent years. Its pathogenesis and pathological process are not yet fully elucidated. The current consensus through multimodal imaging studies is that PEVAC is an idiopathic perifoveal retinal vascular abnormality with cystoid macular edema (CME) and hard exudate. It is not easily distinguished clinically from retinal microaneurysm due to diabetic retinopathy or retinal vein occlusion, aneurysmal telangiectasia, and type 3 macular neovascularization (stage Ⅰ). PEVAC is a partially self-healing property, and some aneurysms and CME may resolve on their own during follow-up period. Laser photocoagulation is an effective measure for the treatment of PEVAC. In the future, with the rapid development of imaging technology, the progress of research on pathogenesis and treatment strategies and the updating of theories are expected to provide more effective treatments for PEVAC.

Objective:To investigate the efficacy and prognostic factors of pars plana vitrectomy (PPV) in the treatment of proliferative diabetic retinopathy (PDR) with chronic renal failure (CRF).Methods:A retrospective study. From January 2016 to June 2021, a total of 82 eyes of 58 patients diagnosed with PDR combined with CRF and treated with PPV in Department of Ophthalmology, The Second Hospital of Hebei Medical University were included in the study. There were 32 cases in males and 26 cases in females. The mean age was (48.45±10.41) years. The course of renal failure was (4.15±3.23) years, and the course of diabetes was (14.45±6.71) years. All patients undergo best-corrected visual acuity (BCVA). The BCVA examination was performed using the international standard Snellen visual acuity chart, which was converted to logarithm of the minimum angle of resolution (logMAR) visual acuity for recording. The mean number of logMAR BCVA was 2.04±0.82 (0.7-2.8). The duration of vitreous hemorrhage averaged (2.65±1.55) months. There were 38 eyes (46.3%, 38/82) with traction retinal detachment; 32 eyes had a history of panretinal photocoagulation (PRP) treatment (39.0%, 32/82). All eyes were treated with 25G PPV. Patients with traction retinal detachment were treated with intravitreal injection of anti-vascular endothelial growth factor (VEGF) 3 days before surgery. Opacification of the lens affected the operation operator combined with phacoemulsification. Biochemical indexes such as hemoglobin, glycosylated hemoglobin, albumin, creatinine, uric acid, and alternative treatment (non-dialysis/hemodialysis/peritoneal dialysis) were collected. Postoperative follow-up time was ≥6 months. χ2 test or Fisher's exact test were used for comparison between groups. A logistic regression model was used for multivariate analysis, and Spearman correlation analysis was used to evaluate the correlations between variables. Results:At 6 months after surgery, the mean logMAR BCVA was 1.16±0.57. Compared with logMAR BCVA before surgery, the difference was statistically significant ( t=-0.837, P<0.001); 44 eyes had BCVA ≥0.1 and 38 eyes had BCVA <0.1. Postoperative vitreous hemorrhage (PVH) was observed in 17 eyes after surgery (20.7%, 17/82). PVH occurred in 15 (46.9%, 15/32), 1 (2.3%, 1/44), and 1 (16.7%, 1/6) eyes in patients without dialysis, hemodialysis and peritoneal dialysis, respectively. There was significant difference between those without dialysis and those on hemodialysis ( χ2=26.506, P<0.05). There was no significant difference between peritoneal dialysis patients and those without dialysis and hemodialysis patients ( χ2=2.694, 2.849; P>0.05). PVH occurred in 3 (10.0%, 3/30) and 14 (27.0%, 14/52) eyes of vitreous cavity filled with silicone oil and perfusion fluid, respectively. The difference was statistically significant ( χ2=3.315, P<0.05); 1 (33.3%, 1/3) and 10 (71.4%, 10/14) eyes were treated with PPV again, respectively, and the difference was statistically significant ( P<0.05). Neovascular glaucoma (NVG) occurred in 12 eyes (14.6%, 12/82). Logistic regression analysis showed that age [odds ratio ( OR) =0.911, P<0.05], diabetic retinopathy (DR) stage ( OR=7.229, P<0.05), renal failure duration ( OR=0.850, P<0.05), operation time ( OR=1.135, P<0.05) was an independent risk factor for poor vision prognosis. Diabetes duration ( OR=1.158, P<0.05), renal failure duration ( OR=1.172, P<0.05) and alternative therapy were independent factors affecting the occurrence of PVH. Diabetes duration ( OR=1.138, P<0.05) and renal failure duration ( OR=1.157, P<0.05) were independent risk factors for postoperative NVG. Spearman correlation analysis showed that PVH was strongly correlated with post-operative NVG ( r=0.469, P<0.01). There was no significant correlation between blood glucose, hemoglobin, creatinine and blood urea nitrogen and prognosis of postoperative vision, PVH and NVG occurrence ( P>0.05). Conclusions:In PDR patients with CRF, DR Stage, age, renal failure course and operation duration are correlated with vision prognosis. Compared with those who do not receive alternative therapy, hemodialysis treatment can reduce the occurrence of PVH and NVG after surgery.