ABSTRACT
AIM: To explore the mechanism of fructus lycii in treating dry eye based on network pharmacology and experimental verification.METHODS: Taking “fructus lycii” as key words, the active ingredients and target of fructus lycii were searched by using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Gene targets related to dry eye(DE)were searched by GeneCards and OMIM databases. The target genes of fructus lycii and DE were imported into Venn software to obtain the intersection target map of them. After that, the data were imported into the String database to obtain the PPI protein-protein interaction network diagram. Using Cytoscape3.7.2 software, the PPI protein-protein interaction network diagram was constructed for active ingredients, target sites and related diseases of fructus lycii. The Bioconductor platform and R language were used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis. And the key targets in the pathogenesis of DE were verified by experiments.RESULTS: Through TCMSP, 45 types of effective chemical components of fructus lycii, 174 target genes corresponding to active components and 131 common target genes with DE were screenedout. In accordance with the network topology of “drug-composition-disease-target”, 27 main effective components of fructus lycii were found in the treatment of DE. The PPI network was analyzed according to the high degree value, which is the key targets of fructus lycii for DE treatment, mainly including AKT1, VEGFA, CASP3, IL1B, JUN, PTGS2, CXCL8, etc. According to GO enrichment analysis, 166 biological functions and processes of fructus lycii for DE treatment were obtained. KEGG enrichment analysis showed that 31 signaling pathways were involved. Additionally, experimental verification displayed that the protein expressions of AKT1, interleukin-6(IL-6), tumor necrosis factor(TNF-α)and IL-17 in conjunctiva tissue of the DE model group were significantly increased.CONCLUSIONS: Through network pharmacology, this study confirmed that the treatment of DE by fructus lycii is a complex process involving multi-components, multi-targets and multi-pathways, and that the treatment of DE by fructus lycii is mainly regulated by anti-inflammatory and apoptosis-related molecules.
ABSTRACT
AIM: To study the effects of the specific simulated luminous environment on the visual performance of people with different vision, so as to provide an experimental basis for revising pilots' vision standards. METHODS: A controlled randomized trial was conducted. Twenty-four volunteers were recruited and divided into four groups(1.0/1.0, 0.8/0.8, 0.6/0.6 and 0.4/0.4, decimal vision)according to right/left eye visual acuity, with six subjects in each group. Each subject was tested for static distant vision, kinetic visual acuity, color vision, depth perception error and visual search time under the simulated luminous environments of sunlight, twilight, and on-cloud, respectively, to compare changes in the impact of distinctive luminous surroundings on the visual performance indicators of human beings with different vision.RESULTS: There were main effect differences in static distant vision, kinetic visual acuity, color error, depth perception error and visual search time under different light environments(all P<0.01). The binocular static distant visual acuity, abilities of color discrimination, depth perception and visual search in simulated sunlight environment were higher than those in simulated twilight and on-cloud environments. In the 0.4/0.4 vision group, kinetic vision in simulated twilight and on-cloud environments were significantly lower than that in simulated sunlight environment(P<0.01). There were main effect differences in binocular static distant vision, kinetic visual acuity, depth perception error and visual search time among subjects with different vision(all P<0.05). Compared with 1.0/1.0 vision group, those with 0.6/0.6 and 0.4/0.4 vision had significant decrease in kinetic visual acuity, depth perception ability and visual search ability(all P<0.05). CONCLUSION: Different luminous environments have a great impact on the visual performance of people with low vision, which poses a potential threat to flight safety.
ABSTRACT
AIM:To investigate the natural course and adverse event of branch retinal vein occlusion (BRVO) rat model induced by laser photochemical method. METHODS: Thirty SD (Sprague Dawley) rats were administrated Bangladesh via tail vein. Then 532nm laser (80mW, 100μ m and 100ms) was performed on retinal vein secondary bifurcation of bitamporal optic disk for 50 spots. Electroretinogram (ERG), fundus fluorescein angiography ( FFA), optical coherence tomography (OCT) and fundus (fluorescein) photograph were applied on 1, 3, 5, 7, 10, 14 and 21d after BRVO model constructed. Two rats were sacrificed, respectively, on 1, 5 and 21d after photocoagulation to carry on HE (Hematoxylin - Eosin stain) and VEGF - α (vascular endothelial growth factor - α) immumohistochemical staining. RESULTS: There were three rats died, three rats with severe retinal detachment for excessive bleeding,one rat with retinal sunken, and one rat with cataract. FFA and fundus ( fluorescein) photograph showed that the successful BRVO rat model was 73% (22/30). It was found that the near-end photocoagulation vein became coarse, far - end became diminution on 1d and the photocoagulation vein total recanalization was on 3-7d. ERG showed the amplification of b wave (dark -adaptation 3.0 response) decreased to 0.694士0.042 of control eyes and on 5-7d decreased to rock bottom about 0.487士0.064 of control eyes. Then it increased Aii the time to 0.708士0.0465 of control eyes on 21d. OCT and HE staining found that retinal ganglion cells and outer nuclear layer became edema on 1d in vivo and in vitro.It was observed that the thickness of retina on photocoagulation vein (0μ m or 250μ m) decreased from 5d and there were 3-4 layer cells in ONL on 21d. The expression of VEGF-α at injured site were significantly more than control eyes on 1d and there were no significant difference on 5d;But the expression of VEGF-α were slightly less than control eyes on 21d. CONCLUSION: Photochemical method was a feasible method to establish BRVO rat model. The evolution and development of the BRVO model could partly mimic human BRVO phenomenon. At the same time, it should be improved to increase the successful model rate.
ABSTRACT
?AIM:To establish a suitable normative reference value for electroretinogram ( ERG ) testing using electroencephalogram ( EEG) skin electrodes.?METHODS: The ERG was recorded in 51 eyes ( 30 people ) who were normal after ophthalmologic examination in our department from March to September 2015 using skin electrodes and contact lens electrodes. The recorded result was reviewed and analyzed, and all the testings were recorded by the routine program.?RESULTS: The 95% confidential interval, mean or median of values was defined in amplitudes and latencies of various responses. All the amplitudes results of skin electrodes were significantly lower than those of the contact lens electrodes and the ratio ( amplitudes of skin electrodes to those of the contact lens electrodes ) was 20% to 30%. The latencies results of skin electrodes were significantly shorter than those of contact lens electrodes and the ratio was 95% to 96%.?CONCLUSION:The EEG skin electrode used for patients with low compliance may provide valuable information of retinal function.
ABSTRACT
Background Light leads to the damage of retinal function and structure by promoting the reproduction of radicals and lipid peroxides when retina is exposed to an intense light environment for a long time.It is necessary to study how to protect the retina against light-induced injury in ophthalmology.Objective The aim of this study was to evaluate the effect of anthocyanidin extracts in preventing retinal photic damage.Methods Eighteen clean male Sprague-Dawley (SD) rats were randomly assigned to normal saline group,anthocyanidin group and mixed (hydrogen-rich saline (HRS) + anthocyanidin,1:1 in volume) group and 6 rats for each.The electroretinogram (ERG) with international standardized 5 items was recorded from all the rats before experiment.Normal saline,anthocyanidin extracts or mixed solution of 5 ml/kg were intraperitoneally injected in the three groups,respectively,for consecutive 5 days.Then the diffused light with the luminance intensity of (5000±300)lx was used to irradiate the right eyes of the rats for consecutive 3 hours during 19:00 through 7:00 in a device made by our laboratory,and the left eyes were covered at the same time.The ERG was repeatedly recorded 5 days after light irradiation.The rats were sacrificed at the end of the experiment and retinal sections were prepared for the histopathological examination.The functional and structural changes of retinas were compared among the different groups.The use of the rats followed the Statement of ARVO.Results The differences of rat body weight were not statistically significant among the three groups (before experiment:F =0.472,P =0.841 ; after experiment:F =0.658,P=0.762).No any significant difference was found in scotopic 0.01 ERG b wave,scotopic 3.0 ERG a and b waves,scotopic 3.0 oscillatory potentials,photopic 3.0 ERG b wave and 3.0 flicker P1wave between the left eyes and the right eyes in the three groups before experiment (P＞0.05).The amplitudes of various waves of ERG were significantly declined in the right eyes compared with the left eyes (P＜0.05).The mean differential values of scotopic 0.01 ERG b wave,scotopic 3.0 ERG a and b waves,scotopic 3.0 oscillatory potentials were significantly different(F =4.594,P=0.029; F=3.834,P=0.037; F=12.823,P=0.000; F=3.976,P=0.032),but those in photopic 3.0 ERG b wave were not statistically significant (F =1.488,P =0.259).Compared with the normal saline group,the differential values of scotopic 0.01 ERG b wave,scotopic 3.0 ERG a and b waves,scotopic 3.0 oscillatory potentials were all reduced in the anthocyanidin group and mixed group (P ＜ 0.05).The cells of outer nuclear layer were decreased in the right eyes in the three groups compared with the left eyes,especially around the optic nerve head and the upside of the retina,with significant differences between them (P＜0.05),and those in the anthocyanidin group and mixed group were significantly less than normal saline group (P＜0.05).Conclusions Anthocyanidin has a protective effect on light-induced retinal damage of SD rats.The protective effect of anthocyanidin extracts is similar to the integrated effect of the mixed group.
ABSTRACT
Background Pupillary light reflex has been widely used in the diagnosis and evaluation of visual system and nervous system diseases.However,in animal experiments,functional evaluation of the visual system and nervous system needs more advanced technology and are affected by many factors.Objective This study was to explore the use of the dynamic pupillometer in evaluating pupillary light reflex and to discuss the influence of brightness of stimulate on relevant curve parameters in C57BL/6 mouse.Methods Ten healthy SPF male C57BL/6 mice were collected in this experiment.White light of five luminance levels (2,8,32,128,256 cd/m2) was used to stimulate the mice following a 2-hour dark adaptation.The stimulation was given at the 60-second intervals,for a duration of 100 ms at every stimulation.An infrared camera and video capture card were used to capture digital images during the measuring process in a scotopic environment,at a speed of 60 frames per second.Measuring outcome was saved in the*.AVI format.A software that was developed by our group was used to determine pupil diameter and output pupillary light reflex curve offline.Pupil initial diameter (R1),constriction amplitude (CA),constriction velocity (CV),latency (T1),time for maximum velocity (T2),time for maximum constriction (T3),time for maximun con-striction to 10.1％ R1 re-dilation (RT)and re-dilation velocity (RV)were assessed,and the correlations between luminosity and measuring parameters were analyzed using the Spearman rank correlation.The use of animals followed the Regulations for thd Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results R1 values showed no statistically significant difference among the 5 different luminosity groups(F=1.117,P=0.361).A positive linear correlation was found between stimulating luminosity and CA(r=0.508,P＜ 0.01),but negative correlations were seen between stimulating luminosity and CV or RV (r=-0.625,-0.609,P＜0.01).T1 and T2 values in the 5 different luminosity groups were not statistically significant (F =0.202,P =0.936 ; F =1.584,P =0.195).The different levels of stimulating luminosity showed positive linear correlations with T3 and RT values (r =0.791,0.609,P＜ 0.01).Conclusions The dynamic pupillometer can quantitatively measure the pupillary light reflex of C57BL/6 mice.The pupillary light reflex dynamic curve parameters of mouse were affected by stimulus luminosity levels.These outcomes offer a basis for the application of the dynamic pupillometer system for measuring pupillary light reflex in animal models.
ABSTRACT
BackgroundOscillatory potentials (OPs) has been used extensively in experimental research and clinical diagnosis,but it is well known that OPs are the summating potentials originated from retinal rod and cone.To separate the rod and cone OPs is helpful for us to diagnose some retinal diseases.ObjectiveThis study was to analyze the characteristics of cone-and rod-driven OPs. Methods The retinal cone degeneration ( RCD ) and congenital stationary night blindness(CSNB) rats were used in this study and SD rats served as control,and 6 rats for each kinds of animals.Scotopic and photopic OPs were recorded in each rat under the dark adaptation for 12 hours and light adaptation for 10 minutes at the stimulate light intensities of -35,-25,-15,-5,0,5 db respectively with RETIscan Visual Electrophysical System.The scotopic and photopic OPs were extracted from flash electroretinogram (FERG) with Maflab7.0 Butterworth filtering waves,and the frequency spectrum of the OPs was analyzed by fast Fourier transform.The characteristics of OPs from RCD rats and CSNB rats were assessed and compared.Results The a wave and b wave of ERG were showed under the dark adaptation condition in both SD and RCD rats,but the b wave was absent in CSNB rat.Under the light adaptation condition,b wave was seen in both SD and CSNB rats,but a wave and b wave of RCD rat were absent.Two peaks were exhibited in both SD and RCD rats under the darkadaptation condition and high intensity of stimulate light at the lower frequency( domain frequency) of 75-110 Hz,90-120 Hz and high frequency ( minor frequency) 90- 120 Hz,110- 135 Hz respectively.In various intensities of stimulate light,CSNB rats appeared a peak at 70-100 Hz.But in light-adaptation and various intensities of stimulate light,the frequency peaks were seen at 75-95 Hz and 70-85 Hz from both SD and CSNB rats respectively.However,under the light adaptation condition,only one peak was seen in SD and CSNB rats at the 75-95 Hz and 70-85 Hz respectively.Compared with SD rats,the mean implied time of b wave was delayed and the amplitude was lowed under the light adaptation (P＜0.05 ),however,no significant differences were found in the implied time and amplitude of b wave of scotopic ERG between SD rats and RCD rats( P＞0.05 ).The scotopic OPs showed the prolong implied time and depressed amplitudes in RCD rats and CSNB rats compared with SD rats( P＜0.05 ),and the photopic OPs presented the prolong implied time and lowed amplitude in CSNB rat in comparison with SD rats (P ＜ O.05 ).ConclusionsCone- and rod-driven OPs appear very different characteristics.The results of this study imply that rod pathway gives more contribution to OPs than cone pathway.Analysis of these results will be helpful for the exploration of the origin of OPs and the diagnosis of the related disease.
ABSTRACT
Some studies suggest that the calcium channels and rennin-angiotensin system (RAS) play pivotal roles in the region-specific vascular adaptation due to simulated weightlessness. This study was designed to clarify if angiotensin II (Ang II) was involved in the adaptational change of the L-type calcium channel (Ca(L)) in the cerebral arterial vascular smooth muscle cells (VSMCs) under simulated weightlessness. Tail suspension (SUS) for 3 d was used to simulate immediate early cardiovascular changes to weightlessness. Then VSMCs in cerebral basilar artery were enzymatically isolated using papain, and Ca(L) current (barium instead of calcium as current carrier) in VSMCs was measured by whole-cell patch-clamp techniques. The results showed that 3-day simulated weightlessness significantly increased current density of Ca(L). However, I-V relationships of normalized peak current densities and steady-state activation curves of Ca(L) were not affected by simulated weightlessness. Although Ang II significantly increased current densities of Ca(L) in both SUS and control rats, the increase of Ca(L) current density in SUS rats was much more than that in control rats. These results suggest that 3-day simulated weightlessness induces the adaptational change of Ca(L) in cerebral VSMCs including increased response to Ang II, indicating that Ang II may play an important role in the adaptational change of cerebral arteries under microgravity.
Subject(s)
Animals , Male , Rats , Adaptation, Physiological , Angiotensin II , Physiology , Calcium Channels, L-Type , Physiology , Cerebral Arteries , Cell Biology , Physiology , Hindlimb Suspension , Muscle, Smooth, Vascular , Cell Biology , Physiology , Myocytes, Smooth Muscle , Metabolism , Physiology , Rats, Sprague-Dawley , Time Factors , Weightlessness SimulationABSTRACT
Background Light-induced retinal damage models vary as many influence factors,herein the modeling method is difficult to copy.It is necessary to establish the grading standardization of retinal damage after retinal light exposure.Objective This study was to improve the modeling method and establish a grading standardization for light-induced retinal damage in rat.Methods Twenty-four SPF 8-10 week-old male SD rats were randomly divided into 4 groups and 6 eyes for each group.The rats were exposed to light intense of 5000 lx for 1,2,3 hours respectively in 3 groups,and other 6 rats served as the normal group.Full-field light exposure experiment was performed for each individual rat separately,and an annular illumination box was used tO ensure the experimental rat moving in a single direction and exposing the right eye in 5000 lx light surrounding during experimental duration.Ganzfeid electroretinogram(ERG)was recorded from the experimental rats at the fifth day after light exposure,and the animals were then sacrificed for histopathology observation to evaluate the retinal thickness change.All procedures which involved animals adhered to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.Results After exposing to intensity light for 1,2,3 hours,the b-wave amplitudes of rod response,maximal mixed response,oscillatory potential in scotopic ERG as well as cone response,20 Hz flicker response of photopic ERG were significant declined as lapse of light exposure time(F=71.690,P=0.000；F=56.250,P=0.000；F=23.610,P=0.000：F=27.130,P=0.000；F=27.030,P=0.000)and lowed by 26.2％,52.5％,70.7％,24.4％,39.3％,58.1％respectively at the end of experiment.Meanwhile,the b-wave latencies of rod response,maximal mixed response in scotopic ERG as well as cone response of photopic ERG were evidently different among different groups (F=1.370,P=0.282；F：0.800,P=0.508；F=11.840,P=0.000；F=2.080,P=0.136).Light induced retinal damage located mainly at the temporal retina area.After intensity light exposure for 1,2,3 hours,the thickness of outer nuclear layer at the superior temporal retina attenuated by 11.3％,25.6％and 72.5％,respectively(P<0.05).A significant difference was seen in mean thickness of outer nuclear layer at superior temporal retina among different groups(F=410.27,P=0.000). Conclusion A standardized grading method for light-induced retinal damage is recommended.The continuous illumination in a intensity of 5000 Ix for 1,2,3 hours can induce the mild,moderate or severe retinal damage respectively at temporal retina.