Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are...Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are limited by the opacity of brain tissue.In this study,we used an innovative tissue cleaning technique combined with retrograde trans-synaptic viral tracing to observe changes in retinorecipient regions connected to retinal ganglion cells in mice after optic nerve injury.Specifically,we performed light-sheet imaging of whole brain tissue after a clearing process.We found that pseudorabies virus 724(PRV724)mostly infected retinal ganglion cells,and that we could use it to retrogradely trace the retinorecipient regions in whole tissue-cleared brains.Unexpectedly,PRV724-traced neurons were more widely distributed compared with data from previous studies.We found that optic nerve injury could selectively modify projections from retinal ganglion cells in the hypothalamic paraventricular nucleus,intergeniculate leaflet,ventral lateral geniculate nucleus,central amygdala,basolateral amygdala,Edinger-Westphal nucleus,and oculomotor nucleus,but not the superior vestibular nucleus,red nucleus,locus coeruleus,gigantocellular reticular nucleus,or facial nerve nucleus.Our findings demonstrate that the tissue clearing technique,combined with retrograde trans-synaptic viral tracing,can be used to objectively and comprehensively evaluate changes in mouse retinorecipient regions that receive projections from retinal ganglion cells after optic nerve injury.Thus,our approach may be useful for future estimations of optic nerve injury and regeneration.展开更多
AIM:To quantitatively evaluate the effect of the combined use of 577-nm subthreshold micropulse macular laser(SML)and multi-point mode pan retinal laser photocoagulation(PRP)on severe non-proliferative diabetic retino...AIM:To quantitatively evaluate the effect of the combined use of 577-nm subthreshold micropulse macular laser(SML)and multi-point mode pan retinal laser photocoagulation(PRP)on severe non-proliferative diabetic retinopathy(NPDR)with central-involved diabetic macular edema(CIDME)using optical coherence tomography angiography(OCTA).METHODS:In this observational clinical study,86 eyes of 86 NPDR patients with CIDME who underwent SML and PRP treatment were included.Images were obtained 1 d before laser and post-laser(1 d,1 wk,1,3,and 6 mo)using AngioV ue software 2.0.Best corrected visual acuity(BCVA,LogM AR),foveal avascular zone area(FAZ),choriocapillary flow area(Ch F),parafoveal vessel density(PVD),capillary density inside disc(CDD),peripapillary capillary density(PCD),macular ganglion cell complex thickness(m GCCT),central macular thickness(CMT),and subfoveal choroidal thickness(ChT)were compared between pre-and post-laser treatment.RESULTS:BCVA remained stable during 6 mo postlaser therapy(pre-laser vs 6 mo post-laser:0.53±0.21 vs 0.5±0.15,P>0.05).PVD,ChF,ChT,CMT,and mGCCT significantly increased 1 d post-laser therapy[pre-laser vs 1 d post-laser:superficial PVD(%),40.51±3.42 vs 42.43±4.68;deep PVD(%),42.66±3.67 vs 44.78±4.52;ChF,1.72±0.21 vs 1.9±0.12 mm^2;ChT,302.45±69.74 vs 319.38±70.93μm;CMT,301.65±110.78 vs 320.86±105.62μm;m GCCT,105.71±10.72 vs 115.46±9.64μm;P<0.05].However,PVD,ChF and ChT decreased to less than baseline level at 6 mo postlaser therapy(pre-laser vs 6 mo post-laser:superficial PVD(%),40.51±3.42 vs 36.32±4.19;deep PVD(%),42.66±3.67 vs 38.76±3.74;Ch F,1.72±0.21 vs 1.62±0.09 mm^2;Ch T,302.45±69.74 vs 289.61±67.55μm;P<0.05),whereas CMT and mG CCT decreased to baseline level at 6 mo postlaser therapy(CMT,301.65±110.78 vs 297.77±90.23μm;m GCCT,105.71±10.72 vs 107.05±11.81μm;P>0.05).Moreover,FAZ continuously increased while CDD and PCD continuously decreased in 6 mo after laser therapy.CMT and ChT had a significant positive correlation with ChF and PVD in most post-laser stages.CONCLUSION:During a 6-month follow-up period after combined use of SML and PRP therapy,BCVA remained stable and there was a decreased trend in macular edema.Blood flow increased at 1 d post-laser therapy and reduced at 6 mo post-laser therapy.展开更多
AIM:To investigate the effects of curcumin(Cur)nanoparticles loaded with chitosan derivatives grafted by deoxycholic acid(Chit-DC)on human retinal pigment epithelial(h RPE)cell proliferation and vascular endothelial g...AIM:To investigate the effects of curcumin(Cur)nanoparticles loaded with chitosan derivatives grafted by deoxycholic acid(Chit-DC)on human retinal pigment epithelial(h RPE)cell proliferation and vascular endothelial growth factor(VEGF)m RNA expression.METHODS:Cur nanoparticles were synthesized with Chit-DC as the carrier and Cur as the supported drug.Cell counting kit-8(CCK-8)method was used to detect the effects of different concentrations of Cur/Chit-DC,Chit-DC,and Cur on the proliferation of h RPE cells for different times.The changes of Cur/Chit-DC and Cur on h RPE cell cycle were determined by flow cytometry.Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the m RNA expression levels of VEGF in h RPE cells treated with Cur,Chit-DC and Cur/Chit-DC at 10μg/m L for 24 h.RESULTS:Different concentrations of Chit-DC nanoparticle treated h RPE cells had no significant difference in terms of optical density(OD)values compared with the control group at 24 h and 48 h.Moreover,there was no change in the cell morphology under a light microscope.After 24 h treatment with Cur/Chit-DC and Cur,the percentage of G0-G1 phase cells increased and the percentage of S phase cells decreased in all concentration groups.Cur/Chit-DC and Cur in all concentration groups inhibited the proliferation of h RPE cells in a time and dose dependent manner,and reduced the expression level of VEGF m RNA.CONCLUSION:The Cur/Chit-DC nanoparticles can release Cur continuously and have sustained release function.Both Cur/Chit-DC nanoparticles and Cur could inhibit h RPE cells cultured in vitro,and could reduce the expression level of VEGF m RNA in h RPE cells.展开更多
基金supported by the National Natural Science Foundation of China,No.81870655(to MBY)。
文摘Successful establishment of reconnection between retinal ganglion cells and retinorecipient regions in the brain is critical to optic nerve regeneration.However,morphological assessments of retinorecipient regions are limited by the opacity of brain tissue.In this study,we used an innovative tissue cleaning technique combined with retrograde trans-synaptic viral tracing to observe changes in retinorecipient regions connected to retinal ganglion cells in mice after optic nerve injury.Specifically,we performed light-sheet imaging of whole brain tissue after a clearing process.We found that pseudorabies virus 724(PRV724)mostly infected retinal ganglion cells,and that we could use it to retrogradely trace the retinorecipient regions in whole tissue-cleared brains.Unexpectedly,PRV724-traced neurons were more widely distributed compared with data from previous studies.We found that optic nerve injury could selectively modify projections from retinal ganglion cells in the hypothalamic paraventricular nucleus,intergeniculate leaflet,ventral lateral geniculate nucleus,central amygdala,basolateral amygdala,Edinger-Westphal nucleus,and oculomotor nucleus,but not the superior vestibular nucleus,red nucleus,locus coeruleus,gigantocellular reticular nucleus,or facial nerve nucleus.Our findings demonstrate that the tissue clearing technique,combined with retrograde trans-synaptic viral tracing,can be used to objectively and comprehensively evaluate changes in mouse retinorecipient regions that receive projections from retinal ganglion cells after optic nerve injury.Thus,our approach may be useful for future estimations of optic nerve injury and regeneration.
基金Supported by the Natural Science Foundation of Guangdong Province(No.2015A030313019)the Sun Yat-sen Clinical Research Cultivation Project(No.SYS-C-201705)。
文摘AIM:To quantitatively evaluate the effect of the combined use of 577-nm subthreshold micropulse macular laser(SML)and multi-point mode pan retinal laser photocoagulation(PRP)on severe non-proliferative diabetic retinopathy(NPDR)with central-involved diabetic macular edema(CIDME)using optical coherence tomography angiography(OCTA).METHODS:In this observational clinical study,86 eyes of 86 NPDR patients with CIDME who underwent SML and PRP treatment were included.Images were obtained 1 d before laser and post-laser(1 d,1 wk,1,3,and 6 mo)using AngioV ue software 2.0.Best corrected visual acuity(BCVA,LogM AR),foveal avascular zone area(FAZ),choriocapillary flow area(Ch F),parafoveal vessel density(PVD),capillary density inside disc(CDD),peripapillary capillary density(PCD),macular ganglion cell complex thickness(m GCCT),central macular thickness(CMT),and subfoveal choroidal thickness(ChT)were compared between pre-and post-laser treatment.RESULTS:BCVA remained stable during 6 mo postlaser therapy(pre-laser vs 6 mo post-laser:0.53±0.21 vs 0.5±0.15,P>0.05).PVD,ChF,ChT,CMT,and mGCCT significantly increased 1 d post-laser therapy[pre-laser vs 1 d post-laser:superficial PVD(%),40.51±3.42 vs 42.43±4.68;deep PVD(%),42.66±3.67 vs 44.78±4.52;ChF,1.72±0.21 vs 1.9±0.12 mm^2;ChT,302.45±69.74 vs 319.38±70.93μm;CMT,301.65±110.78 vs 320.86±105.62μm;m GCCT,105.71±10.72 vs 115.46±9.64μm;P<0.05].However,PVD,ChF and ChT decreased to less than baseline level at 6 mo postlaser therapy(pre-laser vs 6 mo post-laser:superficial PVD(%),40.51±3.42 vs 36.32±4.19;deep PVD(%),42.66±3.67 vs 38.76±3.74;Ch F,1.72±0.21 vs 1.62±0.09 mm^2;Ch T,302.45±69.74 vs 289.61±67.55μm;P<0.05),whereas CMT and mG CCT decreased to baseline level at 6 mo postlaser therapy(CMT,301.65±110.78 vs 297.77±90.23μm;m GCCT,105.71±10.72 vs 107.05±11.81μm;P>0.05).Moreover,FAZ continuously increased while CDD and PCD continuously decreased in 6 mo after laser therapy.CMT and ChT had a significant positive correlation with ChF and PVD in most post-laser stages.CONCLUSION:During a 6-month follow-up period after combined use of SML and PRP therapy,BCVA remained stable and there was a decreased trend in macular edema.Blood flow increased at 1 d post-laser therapy and reduced at 6 mo post-laser therapy.
基金Supported by Hainan Provincial Natural Science Foundation of China(No.819MS133)。
文摘AIM:To investigate the effects of curcumin(Cur)nanoparticles loaded with chitosan derivatives grafted by deoxycholic acid(Chit-DC)on human retinal pigment epithelial(h RPE)cell proliferation and vascular endothelial growth factor(VEGF)m RNA expression.METHODS:Cur nanoparticles were synthesized with Chit-DC as the carrier and Cur as the supported drug.Cell counting kit-8(CCK-8)method was used to detect the effects of different concentrations of Cur/Chit-DC,Chit-DC,and Cur on the proliferation of h RPE cells for different times.The changes of Cur/Chit-DC and Cur on h RPE cell cycle were determined by flow cytometry.Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the m RNA expression levels of VEGF in h RPE cells treated with Cur,Chit-DC and Cur/Chit-DC at 10μg/m L for 24 h.RESULTS:Different concentrations of Chit-DC nanoparticle treated h RPE cells had no significant difference in terms of optical density(OD)values compared with the control group at 24 h and 48 h.Moreover,there was no change in the cell morphology under a light microscope.After 24 h treatment with Cur/Chit-DC and Cur,the percentage of G0-G1 phase cells increased and the percentage of S phase cells decreased in all concentration groups.Cur/Chit-DC and Cur in all concentration groups inhibited the proliferation of h RPE cells in a time and dose dependent manner,and reduced the expression level of VEGF m RNA.CONCLUSION:The Cur/Chit-DC nanoparticles can release Cur continuously and have sustained release function.Both Cur/Chit-DC nanoparticles and Cur could inhibit h RPE cells cultured in vitro,and could reduce the expression level of VEGF m RNA in h RPE cells.
