Objective To research the choroidal neovascularization (CNV)in TgAPPswePS1 transgenic mice after intensive light exposure injury.Methods Twenty TgAPPswe/PS1 transgenic mice at the age of 6 months were grouped for experiments.The treated groups of 12 mice were treated by a source of 10 000 lux cool full spectrum light for 6 months,12 hours per day;While the control groups of 8 mice were kept in normal conditions.The mice eyes of the experimental group and control group were examined with HE/Toluidin blue staining,the retinal structure was observed,and the number of CNV was counted.The expression of VEGF and Aβ were examined with immunofluorescence on the retinal pigment epithelial (RPE) flat mount.All of the results were quantified and statistically analyzed.Results After treated by 6 months of intensive light exposure in the experimental group,histopathological analysis has found significant loss of outer nuclear layer/photoreceptor out segment and outer plexiform layer as compared with the control group;At the same time,abnormal hypo-and hyper-pigmentation,vacuoles and disruption in the RPE layer,remarkable CNV were found in the experiment group by Toluidin blue staining,and the incidence of CNV was 18.75％.The VEGF expression domenstrated.a diffusive and deposition pattern along the neovessels which showed a significant increase of (6.59 ± 1.14) fold changes as compared with the control group.The difference was statistical significant (P ＜ 0.05).Then the Aβ deposits were positive expressed in the RPE layers after intensive light exposure treatment,and pathological deposition of Aβ in the RPE showed plaque like displayed by confocal Z-stack microscopy,and the drusenoid Aβ deposits were found alone with the neovessels on the RPE flat mount.The deposition of Aβ protein increased with (6.45 ± 2.93) fold changes as compared with the control group,and the difference was statistical significant.Conclusion CNV with degenerative changes in the outer retina can be induced by intensive light exposure in the APPswe/PS1 transgenic mouse.These results suggest that an Alzheimer's transgenic animal model might be an alternative animal model for CNV if combined with intensive light exposure.

Objective To research the functional and structural change in TgAP-PswePS1 transgenic mice after intensive light exposure insult.Methods APPswe/PS1 transgenic mice at the age of 6 months old were grouped for experiments,the transgenic mice were replaced by light insult device for 6 months,while the control mice were kept in normal conditions.After 6 months light exposure,the eyes of control and experimental mice were examined with electroretinography (ERG).The retinal morphology change was investigated with H&E staining.All of the results were quantified and statistically analyzed.Results In the control group,the amplitudes of a and b wave in the rod response were (18.33 ±3.53) μV and (107.58 ± 14.72) μV,while (64.80 ±7.57) μV and (178.76 ± 14.47) μV for the amplitudes of a and b wave in the maximum response;After treated by 6 months of intensive light exposure,in experimental group mice,the amplitudes of a and b wave in the rod response were (17.92 ±4.89) μV and (21.83 ± 5.51) μV;While in the maximum response a striking decrease was detected with a wave (18.23 ±4.44) μV and b wave (24.50 ± 4.49) μV,by compared with control group,the difference were statistical significant (all P ＜ 0.05).Histopathological analysis found significant loss of outer nuclear layer,photoreceptor out segment,whereas controls remained little change in the retina.And the retinal thickness decreased significantly from (181.32 ± 13.47) μm in control group to (102.34 ±9.38) μm after light insults in experimental group,the difference was statistical significant (P =0.017).Conclusion Intensive light exposure can cause the retinal structural and functional disorder in the AP-Pswe/PS1 transgenic mouse.

Objective:To investigate the immune effects of CIK cells induced by Toll like receptor 7 agonist (Tlr7a) instead of IFN-γon killing lymphoma cells in vitro .Methods: Mononuclear cells were isolated from healthy human peripheral blood .CIK were induced by Tlr7a in vitro instead of IFN-γ.Two groups were divided as follows:CIK group,Tlr7a-CIK group.Then the main investigation on immune effects included immune phenotype was detected respectively , and cytotoxicity of the effectors was analyzed .Results: In Tlr7a-CIK group,the amount of CD56+cells was more than CIK group (P<0.05),and the cytotoxicity was also stronger (P<0.05). Conclusion:Tlr7a instead of IFN-γcould promote the immune effects of CIK cells on killing tumor cells in vitro .

BrdU (5-Bromo-2'-deoxyuridine) is usually used to label the mitotic cells as well as to trace reagent in cell transplation. However, BrdU could also exert some side effect on cellular biological characteristics upon inappropriate use. To explore the appropriate concentration of BrdU for labelling retinal progenitor cells (RPCs), we co-cultured Embryonic day (E) 17. 5 RPCs with different concentrations of BrdU, which were 0.2, 1, 5 and 10 micromol/L, respectively. After 48 hours, the RPCs were proliferation- or differentiation-cultured. Immunofluorescence was used to detect the BrdU-positive ratio and differentiation potential. Cell count was used to evaluate proliferation ability, and lactate dehydrogenase (LDH) release assay was used to monitor cytotoxicity. The results showed that 0.2 micromol/ L BrdU could not label RPCs clearly, while BrdU of 1, 5 or 10 micromol/L could label the RPCs with similar ratios. 1 micromol/L BrdU displayed no obvious cytotoxicity and showed no obvious effect on the proliferation and differentiation ability. However, 5 micromol/L or 10 micromol/L BrdU could evidently inhibit RPCs proliferation, partly due to the cytotoxicity effect. Furthermore, 10 micromol/L BrdU could inhibit the differentiation of RPCs towards MAP2-positive nerve cells, but showed no influence on the differentiation of RPCs towards GFAP- and glutamine synthetase positive glial cells. This study suggested that 1 micromol/L BrdU could be an appropriate concentration for RPCs labelling and could efficiently label RPCs without obvious side effect.
Animals ; Bromodeoxyuridine ; chemistry ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Proliferation ; Embryo, Mammalian ; Immunohistochemistry ; Mice ; Retina ; cytology ; Staining and Labeling ; Stem Cells ; cytology

Objective To summarize the clinical features in idiopathic hypotony maculopathy.Methods A retrospective case series study.Eighteen eyes of 18 patients who were diagnosed with idiopathic hypotony maculopathy were enrolled in the Second Affiliated Hospital of Wenzhou Medical University from August 2012 to December 2017.There were 8 males (8 eyes) and 10 females (10 eyes).All patients underwent examinations including BCVA,optometry,slit lamp microscope,fundus color photography,UBM,B-mode ultrasound,OCT,FFA and axial length (AL).BCVA was recorded with logMAR acuity.The results of affected eyes and contralateral healthy eyes were compared.Paired t test was performed to compare the intraocular pressure (IOP),diopter and AL of the affected eyes and contralateral healthy eyes.Results Among 18 eyes,there were 6 eyes with logMAR BCVA＜ 1.0,10 eyes with logMAR BCVA 1.0-2.0,2 eyes with light perception.The average diopter was +2.32± 1.78 D.The average IOP was 5.18± 1.38 mmHg (1 mmHg=0.133 kPa).The average AL was 20.92± 1.61 mm.The differences of IOP (t=21.6,P＜ 0.000),diopter (t=5.9,P=0.002) and AL (t=9.13,P＜0.000) between the affected eyes and contralateral healthy eyes were significant.The inflammatory reaction in the anterior segment was observed in 13 eyes (72.22％).In the posterior segment,all the eyes were documented with chorioretinal folds,optic disc swelling and retinal phlebectasia were also demonstrated in 14 eyes,while with macular uplift in 7 eyes.In the UBM and gonioscope examination,the angle chamber was open in all patients with ciliary body cyst in 6 eyes,while no ciliary body detachment was detected.All eyes had been examined with B-scan ultrasound and found the increasing thickness of eye ball in all eyes,and nodular changes in the optic papilla in 5 eyes.The chorioretinal folds were further confirmed by OCT with the appearance of the gear shape,much more obviously in the chomid than that in retina.In the FFA,fluorescein leakage was found around the optic disc in 13 eyes at the late stage,while there was no obvious abnormal leakage in the macular or poster part of retina.Conclusions Idiopathic hypotony maculopathy could present with various clinical manifestations.The choroiretinal folds is typical clinical sign.

Objective To summarize the clinical features and visual outcome of posterior scleritis presented with symptoms involving affected optic nerve.Methods Retrospective case series study.Twelve eyes of 12 female patients with posterior scleritis were included in this study.The average age was 35.2± 14.31 years old.The patients got diagnosed with an average of 24.75 ± 22.91 days.Ocular pain was complained in all patients,and blurred vision in 11 patients.The best corrected visual acuity (BCVA),intraocular pressure (IOP),slit lamp microscope examination,B-scan ultrasound,optical coherence tomography (OCT),fundus photography,fundus fluorescein angiography (FFA) and ocular wall thickness measurement were performed in all patients.Nine eyes received visual field examination.All patients received systemic corticosteroid and steroidal eye drops for 3 months.Clinical features and outcome were retrospectively studied.Results Before treatment,the BCVA was from ＜0.1 to ＞0.8.There were 3 eyes with scleral hyperemia,3 eyes with anterior chamber flares,12 eyes with papilledema and different degrees of retinal vein dilatation,3 eyes with star-shaped macular exudates and 2 eyes with macular retinal pigment epithelium detachment.B-scan ultrasound demonstrated that the ocular walls were thickening in all eyes with typical T-sign,and the average thickness was 2.76± 0.68 mm.OCT demonstrated optic disc swelling,and the macular retinal detachment in 2 eyes.In the FFA examination,the fluorescein leakage of the disc was enhanced with time.In the Humphrey test,the value of mean deviation (MD) was 12.56± 5.73 dB and pattern standard deviation (PSD) was 8.15±4.23 dB in 9 eyes before the treatment.After treatment for 3 months,the symptoms were attenuated and the visual acuity was obviously improved with BCVA＞0.1 in all eyes.Scleral hyperemia and anterior chamber flares were only found in 1 eye.The optic disc edema gradually faded away.The ocular wall thickness in the poster part of the eyeball decreased,and the T-sign disappeared in all eyes,the average thickness was 1.53±0.41 mm.Compared with parameters before the treatment,the difference was statistically significant (t=0.003 5,P＜0.05).OCT demonstrated the recovery of the macular retinal detachment.There was no abnormal leakage evidenced in FFA in the optic disc and macular.After treatment,the value of MD and PSD was 5.19±4.82 dB and (4.33 ±3.76) dB,respectively.The difference of MD value between before and after the treatment was significant (t=0.026,P＜0.05).Conclusions Posterior scleritis with an initial symptom of optic nerve was tend to affect middle-aged patients,with clinical manifestations of anterior segment signs in some patients and optic disc swelling with retinal vein dilatation in all patients.B ultrasound examination showed typical T sign.Systemic corticosteroid treatment always obtained remission of the ocular inflammatory activity,and could achieve favorable visual outcome.