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Objective:To analyze the occurrence of early hypotony after the intravitreal injection of anti-vascular endothelial growth factor (VEGF) and its risk factors.Methods:A case-control study was performed.One hundred and twenty-seven eyes of 127 patients with fundus vascular disease who received intravitreal injections of anti-VEGF drugs were enrolled in Henan Provincial People's Hospital from January 2020 to January 2022.Of the 127 patients, there were 71 males and 56 females, with an average age of (61.85±11.53) years and a mean intraocular pressure of (15.28±3.71)mmHg (1 mmHg=0.133 kPa). All subjects were intravitreally injected with 0.05 ml of anti-VEGF drugs, including 56 cases receiving ranibizumab, 38 cases receiving conbercept and 33 cases receiving aflibercept.The intraocular pressure was measured with a non-contact tonometer at 30 minutes, 1 hour and 2 hours after the injection.The cases were grouped as hypotony group or non-hypotony group according to the intraocular pressure of subjects was less than 10 mmHg or not.The differences in sex, age, distribution of left eye and right eye, disease type, intraocular pressure before injection, injection frequency, lens status, drug type, injection timing, injection site, with or without high myopia, with or without a history of glaucoma or ocular hypertension, and with or without a history of vitreoretinal surgery were analyzed to investigate the factors with a P-value <0.05, which were used as the independent variable and the occurrence of hypotony as the dependent variable in logistic regression analysis to explore the risk factors for hypotony.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEEC-2022-42). Results:Hopotony occurred in 8 eyes within 2 hours after the injection.There were significant differences in intraocular pressure at different time points before and after injection between the hypotony and non-hypotony groups ( Fgroup=62.177, P<0.001; Ftime=25.128, P<0.001). The intraocular pressure of the hypotony group at 30 minutes, 1 hour and 2 hours after injection were lower than before injection, and the intraocular pressure of the non-hypotony group was higher at 30 minutes after injection than before injection (all at P<0.05). The average reduction of intraocular pressure of the hypotony group was 7.88, 7.63 and 7.23 mmHg at 30 minutes, 1 hour and 2 hours after the injection, and the intraocular pressure returned to baseline level at 1 day after injection.There was no significant difference in sex, distribution of left and right eyes, disease type, pre-injection intraocular pressure, injection frequency, lens status, drug type, injection timing, injection site, with or without a history of high myopia and with or without a history of glaucoma or ocular hypertension between the two groups.There were significant differences in age and with or without a history of vitreoretinal surgery between the two groups ( t=8.265, P<0.001; χ2=6.907, P=0.035). Multivariate logistic regression analysis showed younger patients and having a history of vitreoretinal surgery were the risk factors for early hypotony after anti-VEGF intravitreal injection (odds ratio=88.563, P<0.001; odds ratio=20.991, P=0.009). Conclusions:Patients with younger age and having a history of vitreoretinal surgery are susceptible to early hypotony after anti-VEGF intravitreal injection.
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Berberine(BBR)is an isoquinoline alkaloid extracted from Coptis chinensis that improves diabetes,hyperlipidemia and inflammation.Due to the low oral bioavailability of BBR,its mechanism of action is closely related to the gut microbiota.This study focused on the CYP51 enzyme of intestinal bacteria to elucidate a new mechanism of BBR transformation by demethylation in the gut microbiota through multiple analytical techniques.First,the docking of BBR and CYP51 was performed;then,the pharma-cokinetics of BBR was determined in ICR mice in vivo,and the metabolism of BBR in the liver,kidney,gut microbiota and single bacterial strains was examined in vitro.Moreover,16S rRNA analysis of ICR mouse feces indicated the relationship between BBR and the gut microbiota.Finally,recombinant E.coli con-taining cyp51 gene was constructed and the CYP51 enzyme lysate was induced to express.The metabolic characteristics of BBR were analyzed in the CYP51 enzyme lysate system.The results showed that CYP51 in the gut microbiota could bind stably with BBR,and the addition of voriconazole(a specific inhibitor of CYP51)slowed down the metabolism of BBR,which prevented the production of the demethylated metabolites thalifendine and berberrubine.This study demonstrated that CYP51 promoted the deme-thylation of BBR and enhanced its intestinal absorption,providing a new method for studying the metabolic transformation mechanism of isoquinoline alkaloids in vivo.
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Objective:Mutations in the OPN1LW gene located in X chromosome usually lead to blue cone monochromacy.Variations in OPN1LW gene usually occur in the exon region, but was rare in the intron region.This study was to report a Chinese family with X-linked rod-cone dystrophy associated with a novel OPN1LW gene hemizygotic splicing variation and analyze the clinical phenotype and gene mutation characteristics Methods:A pedigree investigation was performed.The family members clinically diagnosed as rod-cone dystrophy with myopia were enrolled in Henan Provincial People's Hospital on January 9, 2020.Detailed ophthalmological examination was carried out, and the periphery venous blood was collected for DNA extraction.The target gene sequencing panel PS400 developed by Henan Clinical Research Center for Eye Diseases and whole exon sequencing were used to detect pathogenic mutations.Sanger sequencing and pedigree co-segregation were used to verify variations.The pathogenicity of the novel variation was analyzed based on American College of Medical Genetics (ACMG) Guidelines and online tools SIFT, Polyphen2, Mutation Taster.This study adhered to the Declaration of Helsinki, and the study protocol was approved by the Medical Ethics Committee of Henan Eye Hospital (HNEECKY-2019 [15]). Written informed consent was obtained from each family member before any medical examination.Results:The proband was a 5-year-old boy with poor vision, red-green blindness and nystagmus in both eyes.No obvious abnormality in ocular anterior segment was found.The boundary of optic disc was clear and the color was reddish, and the reflection of macular fovea was clearly visible.OCT image showed indistinct reflection of some ellipsoids in macular area of both eyes.The amplitudes of a and b waves of full-field ERG were not recorded in scotopic 0.01 scale and significantly reduced in scotopic 3.0 and photopic 3.0 ERG.The uncle of the proband had a more severe clinical phenotype.Wide-angle fundus photography showed high myopia findings, peripheral retinal atrophy and sporadic black lesions, and autofluorescence examination showed attenuated fluorescence in peripheral retina.No obvious abnormality was found in the middle-peripheral retinal region.The results of two kinds next generation sequencing showed a novel hemizygotic splicing variation c. 112+ 2T>G in the intron of OPN1LW gene and an unreported heterozygous variation c. 1913A>C (p.Y638S) in the SEMA4A gene.The c. 112+ 2T>G mutation further leaded to the sequence change of classic splice donor of intron 1.According to the ACMG guidelines, the pathogenicity score was PVS1+ PM2+ PP1, which was considered as a pathogenic level. Conclusions:This is the first report of X-linked rod-cone dystrophy associated with OPN1LW gene variation, and this novel variant c. 112+ 2T>G locates in the intron region.This result is different from past knowledge that variations of OPN1LW gene primarily occur in exon.This study expands the mutational spectrum of OPN1LW gene inducing retinal degeneration and the spectrum of clinical phenotype.
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Objective To investigate the risk factors of postoperative vitreous hemorrhage after minimal vitrectomy without endotamponade for proliferative diabetic retinopathy (PDR).Methods From June 2015 to June 2017,103 eyes of 103 patients with PDR diagnosed and underwent minimalvitrectomy in Henan Provincial People's Hospital were enrolled in the study.There were 58 males and 45 females,with the average age of 58.37± 10.14 years and diabetes duration of 8.7± 7.2 years.Baseline systemic parameters including sex,age,diabetes duration,hypertension,HbA1c,creatinine,whether received anticoagulants,ocular parameters including whether combined with vitreous hemorrhage,whether finished panretinal photocoagulation (PRP),whether received treatment of anti-VEGF,whether combined with iris neovascularization (NVI),lens status preoperatively,whether hypotony postoperatively and intraoperative parameters including whether disc neovascularization (NVD) bleeding,whether fibrovascular membrane (FVM) residual,laser points,whether combined with cataract phacoemulsification were identified by multivariate logistic regression analysis.Results Twenty-nine of 103 eyes (28.15％) developed PVH in 1 day to 6 months after surgery,with self absorption of 18 eyes and reoperation of 11 eyes.Univariate analysis showed there were significant differences in age (t=2.124,P=0.036),anti-VEGF(x2=7.105,P=0.008),NVD bleeding (x2=10.158,P=0.001) and FVM residual(x2=8.445,P=0.004) between patients with and without postoperative vitreous hemorrhage.Sex (x2=0.021,P=0.884),diabetes duration (t=0.87,P=0.386),hypertension (x2=2.004,P=0.157),HbA1c (t=1.211,P=0.229),creatinine (t=0.851,P=0.397),preoperative oral anticoagulants (x2=0.985,P=0.321),preoperative vitreous hemorrhage (x2=0.369,P=0.544),PRP (X2=1.122,P=0.727),NVI (x2=2.635,P=0.105),lens status (x2=0.172,P=0.679),hypotony postoperatively (x2=1.503,P=0.220),laser points (x2=1.391,P=0.238) and combined phacoemulsification surgery (x2=0.458,P=0.499) were not associated with PVH.Multivariate logistic regression analysis revealed the more PVH appeared in younger (OR=1.065,P=0.009) and NVD bleeding (OR=6.048,P=0.001) patients.Conclusion Younger age and NVD bleeding are the important risk factors for PVH after minimal vitrectomy without endotamponade in PDR.
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Objective To observe the clinical effect of minimally invasive vitreoretinal (MIV) surgery combined with a modified suprachoroidal drainage surgery for retinal detachment associated with choroidal detachment (RRDCD).Methods A prospective clinical study.A total of 27 patients (27eyes) diagnosed as RRDCD were recruited in this study.There were 16 males and 11 females,with an average of (53.67± 14.82) years.The mean intraocular pressure (IOP) was (8.2± 2.1) mmHg (1 mmHg=0.133 kPa) and best corrected visual acuity (BCVA) of minimum resolution angle logarithm (logMAR) was 1.87±0.58.All subjects underwent 23G MIV combined a modified suprachoroidal drainage surgery,which 23G stab knife and 1 ml syringe needle were used for surgery.The visual outcome,IOP,rate of retinal reattachment and complications were comparatively analyzed preoperatively and postoperatively.Results At 1 day,10 days,1 month and 3 months after surgery,the average of logMAR BCVA were 1.62 ± 0.67,1.51 ± 0.63,1.39 ± 0.54,1.32± 0.56 and the mean of IOP were (13.47 ± 5.06),(14.43 ± 4.09),(14.89 ± 4.30),(15.38 ± 3.37) mmHg,respectively.There were significant differences of logMAR BCVA and IOP between before and after surgery (F=6.19,15.21;P＜0.05).Retinal reattachments were achieved in 27 eyes (100％) at 1 day and 10 days after surgery.At 1 month and 3 months after surgery,the rate of retinal reattachment were 88.89％ (24 eyes) and 85.19％ (23 eyes),respectively.No severe complications such as endophthalmitis and choroidal hemorrhage were found at follow-up visits.Conclusion MIV combined with a modified suprachoroidal drainage surgery is an effective and safe treatment for RRDCD,which can promote retina tear closure,improve visual acuity.
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Objective To evaluate ocular surface changes following minimal vitreoretinal surgery in postmenopausal women patients with proliferative diabetic retinopathy (PDR).Methods Sixty-one women PDR patients (61 eyes) underwent vitreous microsurgery were recruited in this prospective study,including 31 postmenopausal women (PMW group) and 30 non-postmenopausal women (non-PMW group).The contralateral eyes were considered as the control group.Corneal fluorescein (FL) staining,tear break-up time (TBUT),Schirmer I test (SIT),central corneal sensitivity and ocular surface disease index (OSDI) were estimated.All tests were carried out 1 day preoperatively and 1 day,10 days,1 month and 3 months postoperatively.The student's t test or Mann-Whitney U and ANOVA for repeat measurements test were used.Results Preoperatively,TBUT of surgery and non-surgery eyes in PMW were shorter than non-PMW (t=-2.115,-2.035;P＜0.05),but higher OSDI scores were found in PMW (t=2.482,2.208;P＜ 0.05).TBUT reduction rate (Z=-2.771,-1.993;P＜0.05) and OSDI rising rate (Z=2.539,2.157;P＜0.05) of surgery eyes in PMW were higher than non-PMW 1 day and 10 days postoperatively.The lower SIT of surgery eyes in PMW were observed at 1 day and 10 days (t=-2.403,-2.029;P＜0.05) after surgery.At 10 days after surgery,FL and OSDI scores of surgery eyes in non-PMW returned to preoperative level (Z=-0.447,-0.513;P＞0.05),but in PMW,the recovery process experienced 1 month (Z=-1.500,-0.853;P＞0.05).TBUT and SIT of surgery eyes in two groups both reached preoperative level at 1 month following surgery (Z=-0.715,-1.266,-1.531,-0.522;P＞ 0.05).Conclnsions PMW with PDR had ocular surface dysfunction,which resulted in aggravated dry eye after minimal vitreoretinal surgery.
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Background Leber hereditary optic neuropathy (LHON) is mitochondrial DNA (mtDNA) disease and mainly leads to optical nerve degeneration.Its primary mechanism is synthesis disorder of DN4 protein due to variation of mtDNA 11778 locus.So to construct a vector with exogenous normal ND4 and transfect into mitochondria is a key of gene therapy for LHON.Objective This study was to investigate the in vitro transfection of adeno-associated virus (AAV)-ND4 gene into mitochondria.Methods Human renal epithelial cell lines transfected adenovirus E1A (293 cells) were regularly cultured and divided into two groups.Framework plasmids of recombinant AAV-ND4 or simple AAV2 were added to the cell medium respectively.The expression of ND4 in cells were located 12,24,36 and 48 hours after transfected by Y03 dual fluorescent quantum dots staining.The positive response for ND4 showed the green fluorescence.Results Cultured 293 cells grew well with 80％ confluence.Abundant green fluorescence particles were seen in cytoplasm in the AAV-ND4 transfected group,but only red fluorescence from mitochondrial protein was seen in the simple AAV transfected group under the fluorescence microscope.Conclusions Exogenous ND4 protein can been successfully transfected into mitochondria using the ND4 gene constructed AAV.This result provides experimental evidence for the further study on gene therapy of LHON.
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Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.
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Background The molecular biological mechanisms of diabetic retinopathy(DR)is unclear up to now.Researches have proved that endoplasmic reticulum stress(ER Stress)-associated factors are elevated in peripheral blood in patients with diabetic retinopathy.and P58 IPK can inhibit those factors.So the relationship between P58 IPK and DR is worth to investigate. Objective The aim of this study was to detect the dynamic expression of P58 IPK in the retina of diabetic rats. Methods The diabetic animal models were established in 18 clean male SD rats by intraperitoneal injection of stilptozotiein(STZ)at a dose of 60 mg/kg.The rats were sacrificed in 1,3,6 months after injection.The expression change of P58 IPK mRNA in the rats retina was detected by quantitative real-time RT-PCR.Other 6 matched normal rats were used as control groups.This experiment followed the Regulations for the Administration of Affair Concerning experimental Animals by State Science and Technology Commission. Results The rats showed more drinking,more food and more urine after STZ injection with the blood glucose level≥ 16.5 mmol/L.The success rate of diabetic models was 100%.The A value of P58 IPK mRNA/β-actin in rat retina was 0.800±0.005 and 0.975±0.008 after injection of STZ.and that of control rats was 0.725±0.006,showing statistically significant difference between them(t=22.589,t=62.784,P＜0.05).In 6 months after injection of STZ,the expression of P58 IPK mRNA in experimental diabetic rat retina was evidently lower than the eontrol rats(0.671±0.004 versus 0.725±0.006,t=-17.984,P＜0.05).Conclusion P58 IPK has a close relation to the pathogenesis of DR,and it plays a retarding role for DR.