Background Hypoxia can increase the secretion of vascular endothelial growth factor-A (VEGF A) by retinal pigment epithelium (RPE) cells and initiate choroidal neovascularization (CNV) consequently.Two VEGF-A isoforms,the angiogenic VEGF-A family (VEGFxxx) and the anti-angiogenic VEGF-A family (VEGFxxxb),are found and formed by alternative splicing.However,the expressing changes and its effect in human RPE cells under the normoxia and hypoxia are unclear.Objective This study was to investigate the expression of VEGFxxx b in human RPE cells under hypoxia.Methods ARPE-19,a human RPE cell line,were cultured.CoCl2 150 μmol/L was added into the medium to mimic the hypoxia environment for 0 hour (normoxia group),3,12 and 24 hours to induce the hypoxia cells.The cells or suspension was harvested at various time points.The expressions of VEGFxxx mRNA and VEGFxxxb mRNA in the cells were detected,and the expressions of the total VEGF-A protein and VEGFxxxb protein in the cells were assayed by Western blot.ELISA was used to determine the contents of total VEGF-A and VEGFxxxb proteins in suspension.Results VEGFxxx mRNA and VEGFxxxb mRNA were expressed in the ARPE-19 cells in both normoxia and hypoxia,and multiple VEGFxxx mRNA isoforms appeared,including VEGF121 mRNA,VEGF165 mRNA and extremely faint VEGF189 mRNA,but only VEGF165b band was seen in VEGFxxxb mRNA.With the prolong of hypoxia culture,the expression of VEGFxxx mRNA showed gradual increase,while VEGFxxx b mRNA appeared gradual decrease.Western blot exhibited that VEGFxxxb was expressed in the cells cultured by normoxia and hypoxia with the strongest expression in VEGF165b.The VEGFxxxb content in suspension of medium was (166.82± 2.55) pg/ml under the normoxia environment and (125.35 ±2.10) pg/ml in 24 hours under the hypoxia,and the total VEGF-A protein elevated from (294.27 ± 11.97) pg/ml under the normoxia environment to (582.26 ± 12.98) pg/ml in 24 hours under the hypoxia.In addition,the proportion of VEGFxxxb to total VEGF-A in the cells lowed from (56.71 ± 1.02) ％ under the normoxia environment to (21.53 ±0.08) ％ in 24 hours under the hypoxia.Conclusions VEGFxxxb isoforms are expressed in both normoxia and hypoxia ARPE-19 cells.VEGFxxxb isoform is a predominant isoform in normoxia ARPE-19 cells.In hypoxia ARPE-19 cells,however,the expression of VEGF A is significantly stronger than that of VEGFxxxb.

Objective To study the clinical manifestations and histopathological characteristics of acute ocular injuries caused by the unsymmetrical dimethylhydrazine propellant in rabbits,and to investigate the mechanism of ocular injuries caused by unsymmetrical dimethylhydrazine.Methods A rabbit eye tainting model was established by placing propellant-grade liquid on the corneal surface.The rabbit eyes were divided into the normal control group and the 2 h,1 d,3 d and 7 d post-toxicity groups.The corneal clouding in each group was scored,the integrity of the corneal epithelium was observed,the percentage of corneal epithelial defects was calculated,fundus photographs were taken of the rabbits in each group,and the changes in the posterior segment of the retina of the rabbits were observed.Histopathological slides of rabbit eyes were prepared at each time point.Pathological sections were taken from the rabbit eyes at each time point to observe the histo-pathological changes of the cornea,lens,iris,ciliary body and retina.Immunohistochemistry was used to analyze the expres-sions of CD31,TNF-α and MMP-2 in the tissues of eyes of each group.Results The score of corneal opacity degree in each group was 0,but was 2.00(1.25,2.50),3.00(2.75,3.00),2.00(2.00,2.50),1.00(0.75,1.00)after exposure,so the difference was statistically significant(P<0.001).The percentage of the size of the epithelial defect was(100.00±0.00)%2 hours after exposure to the toxin,but was subsequently(99.17±6.36)%,(17.38±5.66)%,and(9.71±3.00)%,respectively.The difference between the groups was statistically significant(P<0.001).Rabbits in each groups had normal fundi.Histopathological examination found that the corneal injury to the rabbits after exposure to the poison was manifested as loss of epithelia,edema and thickening of the stroma layer,cystic degeneration of epithelial cells,vasodilation and congestion of the iris and ciliary body,mild edema of the stroma layer,swelling of muscle fibers,all of which recovered within 3 days.The lens structure of the rabbits in each group remained unchanged.The expressions of MMP-2 in the cytoplasm of the corneal stroma,roots of theiris and ciliary body,and in endothelial cells in the corneal limbus vascular wall were significantly enhanced 1 day after exposure,so was the expression of CD31 in the corneal stroma and limbus.No positive expression of TNF-αwas found in anyeye tissuein these groups.Conclusion The unsymmetrical dimethylhydra-zine propellant can damage eye tissues under a simulated working environment,but the damage is limited.The damage of unsymmetrical dimethylhydrazine to the cornea is superficial and confined to the corneal epithelium.The damage to the anterior segment is mild,and inflammation has a quick onset and recovery.The damage to the cornea,iris and ciliary body tends to recover on its own.

Objective:To explore the retinopathy knowledge, attitude and behavior of inpatients with diabetes mellitus and their correlation factors and analyzed the influencing factors.Methods:This study was cross-sectional. From December 2018 to June 2019, a total of 392 inpatients with diabetes mellitus at a Class Ⅲ hospital were selected by convenience sampling and their knowledge, attitude, behavior and retinopathy were investigated with the diabetic retinopathy knowledge, attitude and behavior questionnaire and the color fundus photograph.Results:Among 392 diabetic inpatients, 267 diabetic inpatients with retinopathy and 123 of them needed treatment. There were 20.9% (82/392) of patients with high-level of retinopathy knowledge, 10.5% (41/392) of them with the positive attitude and 7.9% (31/392) of them with good behavior. Multiple regression analysis showed that the related factors of the total scores of retinopathy knowledge, attitude and behavior were the retinopathy stage and diabetic course ( P<0.01) ; knowledge related factors were the genders and retinopathy stage ( P<0.01) ; attitude related factors were the treatment methods of diabetes mellitus, retinopathy stage and diabetic course ( P<0.01) ; behavior related factor was the retinopathy stage ( P<0.01) . Conclusions:Inpatients with diabetes mellitus have the low levels of retinopathy knowledge, attitude and behavior which points out that medical staff should pay more attention to the improvement of the retinopathy knowledge, attitude and behavior, especially in the patients with the low levels of diabetic knowledge, family incomes monthly, short diabetic courses and without severe retinopathy. Medical staff should also make diabetic knowledge popular and improve the patients' attitudes with regular fundus examination for diabetic retinopathy to promote the good fundus examination behavior.

A portable washing device for eyes in on-site treatment was designed to be used in emergent event,so as to effectively respond to the needs of injury condition of on-site eye contamination,emergency rescue and other cure.The portable washing device for eyes designed in this study consists of an ophthalmic first-aid washing kit,an ocular electronic cleaning instrument and a washing needle for puncture in anterior chamber.The first-aid washing kit is equipped with a case and a protective cover,and the case is capable of storing spare first-aid medicines and instruments.It is possible to quickly check whether the medicines and instruments are all in readiness in the case through the transparently protective cover of the first-aid washing kit.The simple and convenient operation of this device can realize self-rescue operation of washing both eyes'conjunctival sacs at the same time by single person.This washing device not only can wash conjunctival sac on the contaminated ocular surface,but also can complete cleaning function for poison in the anterior chamber,and can real-timely monitor the acid-base value of the anterior chamber by using the measuring valve in the cavity of the ophthalmic electronic cleaning instrument.This washing device designed in this study is a highly practical toolkit specially designed for dealing with first-aid situations of eyes,which has many advantages include meeting the basic functions of emergency medical rescue for required nuclear and chemical injuries in daily training and support,emergency rescue and disaster relief,as well as defense and service support in wartime,and raising the emphasis of people to first-aid for eyes,and providing timely rescue for eyes.This device has a favorable value of conversion and promotion.

To explore the differentially expressed genes (DEGs) related to biosynthesis of active ingredients in wolfberry fruits of different varieties of Lycium barbarum L. and reveal the molecular mechanism of the differences of active ingredients, we utilized Illumina NovaSeq 6000 high-throughput sequencing technology to conduct transcriptome sequencing on the fruits of 'Ningqi No.1' and 'Ningqi No.7' during the green fruit stage, color turning stage and maturity stage. Subsequently, we compared the profiles of related gene expression in the fruits of the two varieties at different development stages. The results showed that a total of 811 818 178 clean reads were obtained, resulting in 121.76 Gb of valid data. There were 2 827, 2 552 and 2 311 DEGs obtained during the green fruit stage, color turning stage and maturity stage of 'Ningqi No. 1' and 'Ningqi No. 7', respectively, among which 2 153, 2 050 and 1 825 genes were annotated in six databases, including gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and clusters of orthologous groups of proteins (KOG). In GO database, 1 307, 865 and 624 DEGs of green fruit stage, color turning stage and maturity stage were found to be enriched in biological processes, cell components and molecular functions, respectively. In the KEGG database, the DEGs at three developmental stages were mainly concentrated in metabolic pathways, biosynthesis of secondary metabolites and plant-pathogen interaction. In KOG database, 1 775, 1 751 and 1 541 DEGs were annotated at three developmental stages, respectively. Searching the annotated genes against the PubMed database revealed 18, 26 and 24 DEGs related to the synthesis of active ingredients were mined at the green fruit stage, color turning stage and maturity stage, respectively. These genes are involved in carotenoid, flavonoid, terpenoid, alkaloid, vitamin metabolic pathways, etc. Seven DEGs were verified by RT-qPCR, which showed consistent results with transcriptome sequencing. This study provides preliminary evidences for the differences in the content of active ingredients in different Lycium barbarum L. varieties from the transcriptional level. These evidences may facilitate further exploring the key genes for active ingredients biosynthesis in Lycium barbarum L. and analyzing their expression regulation mechanism.
Flavonoids/metabolism* ; Fruit/genetics* ; Gene Expression Profiling/methods* ; Gene Expression Regulation, Plant ; Lycium/metabolism* ; Metabolic Networks and Pathways ; Transcriptome