Background Proliferative vitreoretinopathy (PVR) is a common cause of vision loss clinically,and retinal pigment epithelium (RPE) cells play a major part in this disease.Studying the effect of traditional Chinese medicine on RPE cells are of great importance to reveal the pathogenesis and prevention of PVR,which were rarely reported.Objective This study was to study and compare the inhibition effect among curcumin,salvia miltiorrhiza and matrine on IL-1β-induced proliferation of rabbit RPE cells.Methods RPE cells at passages 3-4 were enrolled for the research and identified by transmission electron microscope.The proliferation effect of IL-1 β (2.5,5.0,10.0,20.0 μg/L) and inhibitory effect of curcumin (5,10,20 μg/ml),salvia miltiorrhiza (5,10,20 μg/ml)or matrine (100,200,400 μg/ml) on RPE cells 24,48 and 72 hours after cultivation were studied by MTT assay.The 50％ inhibitory dose (IC50) of the three medicines were analyzed by regression analysis.The use and feeding of the experimental animals were followed by the ARVO Statement.Results RPE cells isolated from the rabbit eye were in round shape and abundant in melanin;The melanin significantly decreased in the fourth generations of RPE cells.Immunohistochemistry showed that the RPE cells was positive for keratin (AE1/AE3).The proliferation rates of RPE cells were statistically different among different concentrations of IL-1β 24,48 and 72 hours after cultivation (Ftime =30.33,P =0.00;Fconcentration =9.37,P =0.00);The proliferation rates of RPE were significantly different among different time points or different concentrations of IL-1β (all at P ＜ 0.05).And the proliferation rate run up to maximum at 10 μg/L after 72 hours of cultivation.The inhibitory rates of the three medicines were statistically different among different time points or different concentrations (curcumin:Ftime =128.75,P =0.00;Fconcentration =334.05,P=0.00.salvia miltiorrhiza:Ftime =39.32,P=0.00;Fconcentration =165.57,P=0.00.matrine:Ftime =267.76,P =0.00;Fconcentration =912.34,P =0.00).The three medicines dose-dependently and time-dependently inhibit IL-1β-induced proliferation of RPE cells,with significant differences between the adjacent time points and concentrations (all at P＜0.05).The IC50 were 26.77,19.01 and 9.45 μg/ml for curcumin;33.72,23.47 and 12.56 μg/ml for salvia miltiorrhiza,570.96,352.25 and 97.50μg/ml for matrine 24,48 and 72 hours after cultivation.Conclusions The proliferation of RPE cells can be stimulated by IL-1β,and the maximal proliferation occurred with a concentration of 10.0 μg/L IL-1β.Curcumin,salvia miltiorrhiza and matrine dose-dependently and time-dependently inhibit proliferation of RPE cells induced by IL-1β.Curcumin is the best medicine to inhibit the proliferation of RPE cells.

Background Interleukin-1β (IL-1β) is an important inflammation-related factor in the initial stage of proliferative vitreoretinopathy (PVR).The previous research showed that curcumin can inhibit IL-1 β-induced proliferation of rabbit retinal pigment epithelium (RPE) cells,but the anti-inflammatory mechanism and effect of curcumin are still undefined.Objective This study was to observe the migration of IL-1β-induced rabbit RPE cells,and evaluate the function and mechanism of inhibition of curcumin on IL-1β-induced inflammation of RPE cells.Methods Cultured rabbit RPE cells of generation 4 were used in this experiment.The cells were cultured in serum-free DMEM and 0,0.1,1.0 and 10.0 μg/L IL-1β were separately added in the medium for 24 hours.The expressions of cyclooxygenase-2 (COX-2) protein and mRNA in the cells were detected by Western blot and reverse transcription PCR to determine the optimal concentration of IL-1β.The cells were divided into IL-1β group and curcumin+IL-1β group,and 1.0 μg/L IL-1 or 1.0 μμg/L IL-1 β combined with 10 μg/ml curcumin was respectively added into the medium for 24,48 and 72 hours.The cells cultured by only serum-free medium served as the control group.Hematoxylin and eosin staining was conducted for the cells to count the number of cells migrating into the injured area under the optical microscope.The relative expression levels of COX-2 protein and mRNA in the cells were detected by Western blot and reverse transcription PCR,and the relative expression levels of nuclear factor (NF)-κBp65 and inhibitor of NF-κB-α (IκB-α) protein were also detected by Western blot assay.The expression intensity and location of NF-κBp65,IκB-α and COX-2 in the cells were detected by immunochemistry.Results RPE cells just isolated from the rabbit eyes were in round shape and abundant in melanin.The melanin significantly decreased in the fourth generations of RPE cells.The shape of cells became long and narrow,and net shaped distribution.Immunochemistry demonstrated the strong positive response of RPE cells for keratin (AE1/AE3).There were (31.93 ±1.21),(36.27±2.50) and (38.33±2.40) migratory cells in the control group after 24,48 and 72 hours respectively.The number of migratory cells increased to 45.73 ± 2.30,71.13 ± 1.92 and 80.60 ± 1.71 in the IL-13 group,but obviously decreased to 13.13 ± 2.20,14.93 ± 1.10 and 12.60 ± 1.51 in the curcumin + IL-1β group.A Significant increase in the migrating cell number was found in the IL-1 β group compared with the control group and the curcumin+IL-1β group in various time points (all at P＜0.05).The relative expression levels of COX-2 protein and mRNA peaked in the 1.0 μg/L IL-1β group,so 1.0 μg/L of IL-1β was determined as the optimal concentration in the experiment.In 24,48 and 72 hours after culture,the expression levels of COX-2 protein and mRNA in the cells were significantly lower in the curcumin + IL-1β group than those in the control group (all at P＜0.05).The relative expression level reached peak in NF-κBp65 protein and lowed bottom in IκB-α proteins at 48 hours after cultured in the IL-1β group,and the reverse trend was seen in the curcumin+IL-1β group,with the significant differences between the two groups (both at P＜0.05).Immunochemistry showed that NF-κBp65 was expressed strongly in the cell nuclei and cytoplasm in the IL-1 β group and presented the weaker expression in the control group and the curcumin+IL-1 β group.Compared with the control group,the expression was weaker in IκB-α and stronger in COX-2 in the IL-1β group.In addition,the expression of IκB-α was enhanced and that of COX-2 was attenuated in the curcumin+IL-1β group in comparison with the IL-1β group.Conclusions Curcumin inhibits the movement of rabbit RPE cells induced by IL-1β.IL-1β up-regulates the expression of COX-2 by activating NF-κB signal pathway,and curcumin plays an anti-inflammatory role by blocking this pathway.

Objective: To observe the changes of choroidal capillary flow area (CBFA) and density in patients with idiopathic macular hole (IMH) before and after surgery. Methods: A prospective clinical study. Thirty patients (60 eyes) with unilateral IMH diagnosed in Department of Ophthalmology of The Second Hospital of Hebei Medical University from February 2017 to March 2018 and 30 age-and sex- matched normal controls were included in this study. All eyes were divided into group A (30 affected eyes), group B (30 fellow eyes) and group C (30 normal eyes of controls). Among 30 eyes in group A, there were 5, 14, 11 eyes with hole in stage Ⅱ, Ⅲ, Ⅳ, respecitvely. The eyes in group A were divided into two subgroups according to the diameter of the holes: group D: hole diameter less than or equal to 500 μm (11 eyes), group E: hole diameter over than 500 μm (19 eyes). All eyes in group A underwent vitrectomy. Spectral-domain OCT was used to observe the diameter size of macular hole and the closure of the hole after vitrectomy in eyes of group A. The CBFA and blood flow density of superficial choroidal capillaries were measured by OCT angiography before and 3 months after vitrectomy in groups A, B and C. Univariate analysis of variance and t test were used for statistical analysis. Results: Before surgery, in group A, B and C, the CBFA were 2.84±0.35, 3.19±0.23, 3.26±0.24 mm², the blood flow density were (20.74±8.26)%, (35.18±5.20)%, (35.20±6.49)%, respectively. The CBFA and blood flow density in group A were significantly lower than those in group B and C (F=19.768, 45.583; P = 0.000, 0.000), but there was no significant difference between group B and C (F=19.768, 45.583; P=0.332, 0.994). The CBFA and blood flow density in group D were higher than those in group E (t=2.230, 2.202; P=0.034, 0.036). The diameter of macular hole was negatively correlated with CBFA and blood flow density (r=-0.377, -0.477; P=0.044, 0.009). Three months after surgery, the macular holes in group A were closed; CBFA and blood flow density in macular area were significantly higher than before surgery (t=-4.126, -4.912; P=0.000, 0.000). Conclusions CBFA and blood flow density decreased in the macular area of IMH. CBFA and blood flow density can be recovered after vitrectomy.

Objective To investigate the hemodynamics of ocular vessels of the bilateral eyes in patients with unilateral non-arteritic anterior ischemic optic neuropathy(NAION) . Methods Clinical data of 28 patients with unilateral NAION were collected . The peak systolic velocity ( PSV ) ,end diastolic velocity (EDV) ,resistance index (RI) of ophthalmic artery (OA) ,central retinal artery (CRA) ,nasal short posterior ciliary arteries ( NPCA ) and temporal posterior ciliary arteries ( TPCA ) were comparatively analyzed by color Doppler ultrasonography . Results The hemodynamic parameters of OA in NAION were not significantly different from those in the contralateral eyes ( P ＞ 0 .05 ).PSV of CRA was lower in NAION than that in contralateral eyes ,as well as EDV ( P ＜ 0 .05 ) ,while the RI of CRA was not significantly different between two groups ( P ＞0 .05).PSV of NPCA in NAION was lower than that in the contralateral eyes ( P ＜ 0 .05) ,while EDV and RI of NPCA were not quite different from those in contralateral eyes ( P ＞0 .05).The hemodynamic parameters of TPCA in NAION were not significantly different from those in the contralateral eyes ( P ＞0 .05).Conclusions Color Doppler ultrasonography can non-invasively detect and monitor the hemodynamic changes of ocular vessels in NAION ,which will be of great help in illuminating the pathogenesis of NAION and providing groundbreaking evidence for the diagnosis and prognosis of NAION .

Objective:To observe the abnormal clinical manifestations of retinal blood vessels and the characteristic image characteristics of optical coherence tomography (OCT) in young myopia.Methods:A case observation study. From July to December 2020, 523 young patients with different myopia refractive powers who were treated in Department of Ophthalmology of The Second Hospital of Hebei Medical University were included in the study. Among them, 277 were males and 246 were females; the median age was 19.0 (5.0) years. All the affected eyes underwent best corrected visual acuity (BCVA), frequency domain OCT (SD-OCT) examination and axial length (AL) measurement. The BCVA examination was performed using the Snellen eye chart. The median myopia refractive power of the affected eye was 5.00 (3.25) D. Among them, low myopia, moderate myopia, and high myopia were 227, 405, and 414 eyes, respectively. The average AL of the affected eye was 25.6±2.8 mm. The frequency domain OCT instrument was used to scan the temporal side of the retina, the upper and lower nasal vascular arches and the macular fovea radially. The images of retinal vascular cysts, microfolds, and lamellar hole were acquired and stored. The prevalence, composition ratio, distribution rule and OCT imaging characteristics of retinal paravascular abnormalities were observed and analyzed. The distribution of paravascular abnormalities in the retina was compared by the χ2 test; the age, refractive power, and AL of different paravascular abnormalities were compared by the K-W rank sum test. Results:Of the 1046 different diopters of myopic eyes, there were 227 eyes in mild myopia, 405 eyes in moderate myopia and 414 eyes in high myopia. Retinal paravascular abnormalities were detected by SD-OCT in 40 eyes (3.8%,40/1046). The prevalence of retinal paravascular abnormalities in moderate myopia was 0.7% (3/405) and high myopia was 8.9% (37/414). No retinal paravascular abnormalities were observed in mild myopia.Retinal paravascular cysts in 40 eyes (3.8%, 40/1046), retinal paravascular microfolds in 28 eyes (2.7%, 28/1046) and retinal paravascular lamellar holes in 13 eyes (1.2%, 13/1046). Of 40 eyes with retinal paravascular abnormalities, retinal paravascular cysts in all 40 eyes (100.0%, 40/40), retinal paravascular microfolds in 28 eyes (70.0%, 28/40) and retinal paravascular lamellar holes in 13 eyes (32.5%, 13/40). Twelve eyes with simple cyst cavity (30.0%, 12/40); 15 eyes were with cyst cavity with micro-wrinkles (37.5%, 15/40); 13 eyes were with cyst cavity, micro-wrinkles and lamellar holes (32.5%, 13/40). The temporal vascular arch retinal paravascular cysts ( χ2=25.664), microfolds ( χ2=14.973), and lamellar holes ( χ2=13.499) were significantly more than those on the nasal side, and the difference was statistically significant ( P <0.001). Conclusions:The total prevalence of retinal paravascular abnormalities in young myopia is 3.8%; it can occur in both moderate and high myopia. The paravascular cyst may be the earliest pathology of paravascular abnormalities in the retina. The three paravascular abnormalities are mostly distributed along the temporal arch of the retina.

Objective:To analyze the characteristics of eyes with congenital optic disc pits (ODPs) through multimodal imaging.Methods:A cross-sectional study was conducted.Thirty-eight patients (38 eyes) diagnosed with congenital ODPs in the Second Hospital of Hebei Medical University from January 2009 to January 2020 were enrolled.A comprehensive summary analysis of the imaging results including fundus photography, spectral domain-optical coherence tomography (SD-OCT), infrared imaging, fundus autofluorescence (FAF), fluorescein fundus angiography (FFA) and indocyanine green angiography (ICGA) was performed.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of The Second Hospital of Hebei Medical University (No.2021-P011). Written informed consent was obtained from each subject prior to any medical examination.Results:Among the 38 eyes, there were 32 eyes with ODPs located in or below the temporal side of optic disc, 4 eyes with ODPs located above the temporal side of optic disc, and 2 eyes with ODPs located at the center of optic disc, which were round or quasi-circular pale depression, and dark red eminences with clear or unclear boundaries between milk spots were found in 29 eyes with optical-disc macular degeneration (ODP-M) by fundus photography.SD-OCT examination showed that the structure of lamina cribrosa in the lesion area in all ODPs patients was incomplete, which presented a dark area with no tissue reflection, and the fissure led to the deep optic nerve.Fluid was found in the outer nuclear layer in all ODP-M patients, and there were 27 eyes with fluid in the inner nuclear layer, 13 eyes in the ganglion cell layer, and 4 eyes under the inner limiting membrane.Among the 29 eyes with ODP-M, there were 21 eyes with retinoschisis in outer layer, 27 eyes with neuroepithelial detachment.In the 27 eyes with neuroepithelial detachment, spot-like high reflection and reduced or disappeared ellipsoid band reflectance were seen above the neuroepithelium in 18 eyes.In infrared images, there were circular or quasi-circular low-reflection areas in the temporal side of the optic disc, and the lesion of ODP-M eyes presented low-reflection areas.FAF examination showed that in 27 eyes with ODP-M, there was a hypofluorescent region at the posterior pole consistent with the lesion range, among which, there was a granular or sheet-like hyperfluorescence at the center of the hypofluorescent region in 18 eyes.FFA showed that the optic disc depression in the arterial phase of patients was in a localized hypofluorescence state.During the venous phase, fluorescein dye extravasation along the temporal side of the optic disc could be found.A strong fluorescent arc with unclear boundaries at the temporal edge of the optic disc was formed in the late stage of angiography.Among the 29 eyes with ODP-M, the area of the macular lesion showed hyperfluorescence during the late stage of angiography in 27 eyes with neuroepithelial detachment, and no extension of dye toward the macula was found.ICGA showed that the optic disc depression of ODPs patients presented a localized hypofluorescence, and the lesion showed hyperfluorescence in 27 of the 29 ODP-M eyes with neuroepithelial detachment.Conclusions:Multimodal imaging can be helpful to realize the early diagnosis, etiology analysis of ODPs and make treatment plan.

Objective:To evaluate the efficacy and safety of oral anisodine hydrobromide tablets in the treatment of nonarteritic anterior ischemic optic neuropathy (NAION).Methods:A multicenter nonrandomized controlled trial was conducted.A total of 282 acute NAION patients (282 eyes) were recruited from 16 hospitals in China from July 2020 to May 2021.Patients were divided into two groups according to treatment methods, which were control group (124 cases, 124 eyes) receiving regular treatment including citicoline sodium plus Ginkgo biloba leaf liquid extract or Ginkgo biloba leaf extract tablets plus mecobalamin, and experimental group (158 cases, 158 eyes) receiving treatment in control group plus oral anisodine hydrobromide tablets 1 mg, twice daily for 2 to 3 months.Best corrected visual acuity (BCVA), visual field index (VFI), peripapillary retinal nerve fiber layer (pRNFL) and radial peripapillary capillary vessel density (RPC) were assessed at 1, 2, 3, and 6 months after enrollment using the standard decimal visual acuity chart, 750i Humphery visual field analyzer, Cirrus HD-OCT 4000/Cirrus HD-OCT 5000, RTVue-XR optical coherence tomography respectively.The primary outcomes were BCVA and VFI, and the secondary outcomes were pRNFL, RPC, and the side effects during the follow-up.The study adhered to the Declaration of Helsinki.All patients were fully informed about the treatment and purpose of this study and voluntarily signed the informed consent form.The study protocol was approved by Chinese PLA General Hospital (No.S2020-021-01). Results:In all, 242 patients (242 eyes) completed the follow-up of BCVA, and 98 patients (98 eyes) completed the VFI follow-up.In terms of visual function, BCVA and VFI improved significantly over time in the two groups, and BCVA and VFI were better in experimental group than in control group at various follow-up time points (all at P<0.05). In terms of structure, pRNFL gradually decreased in both groups with the extension of treatment, and pRNFL was significanthy thinner in experimental group than in control group at various follow-up time points (all at P<0.05). There was no significant difference in RPC between the two groups at the last follow-up ( P>0.05). There were two cases with side effects and one case was discontinued due to side effects 25 days after enrollment. Conclusions:Oral anisodine hydrobromide can improve visual acuity and visual field in NAION and accelerate the regression of optic disc edema, with good safety.