Preliminary Study on in Vitro Induced Differentiation of Embryonic Stem Cells into Neurons

Purpose:To study preliminarily in vitro induced differentiation of embryonic stem cells into neurons for further investigation of an alternative for the treatment of glaumatous neuropathy.Materials and methods:Supernatant of cultured Buffalo rat liver cells(buffalorat liver cell-conditioned medium,BRL-CM)was used for culturing embryonic stem cells(ES-D3 cell line),Morphological features of undifferentiated EScells were studied by HE staining and electron microscopy.Based on the methods used by Bain et al,we modified the methods and used retinoic acid(RA)as an inducer to differentiate ES-D3 cells and cytosine arabinoside(Ara-C)as inhibitor of proliferative cells.The growth of the cells was observed under phase contrast microscope.Results:ES-D3cells cultured by BRL-CMgrew in aggregates and remained undifferentiated.Electromicroscopy showed large nucleus and a large amount of mitochondria in undifferentiated ES cells and many processes on the surfaces.In the first day after the adding of retinoic acid,some neuron-like cells with one,two or more processes were present.In the second day after adding RAand the first day after the plus of 10μm Ara-C,a large amount of neuron-like cells appeared,with the formation of neuron-like networks.Con clusions:Combined use of RA and Ara-Ccan induce ES cells to differentiate into neuron-like cells.Our present preliminary study might provide insights into an alternative for the treatment of glaucomatous neuropathy by the transplantation of embryonic stem cells.Eye Science2000;16:1-6.

Characteristics of Optic Nerve Damage Induced by Chronic Intraocular Hypertension in Rat

Purpose:To set up the Sharma’s chronic intraocular hypertension model and investigate the intraocular pressure (IOP) as well as the optic nerve damage of this model in rat. Methods:The operations of the chronic intraocular hypertension model were performed as described by Sharma in 60 male Lewis albino rats. IOP was measured using the Tono-Pen XL immediately after surgery and then at 5 day, 2 week or 4 week intervals. Cresyl violet staining of whole-mounted retinas was used to label retinal ganglion cells (RGCs), then RGCs were counted. Paraphenylenediamine (PPD) staining was performed in the semi-thin cross sections of optic nerve of rat, in order to know whether the axons of optic nerve were degenerated or not. Results:There were 47 rats with higher IOP after the episcleral veins cauterized in 60 rats. The ratio of elevated IOP was 78.3%. The IOPs were stable in 4 weeks. After cresyl violet staining, the RGCs loss was 11.0% and 11.3% was found in the central and peripheral retina respectively after 2 weeks of increased IOP. After 4 weeks of increased IOP, the loss of RGCs was 17% for the central retina and 24.6% for the peripheral retina. In the retinas without higher IOP, there was no loss of RGCs. PPD staining showed that optic nerve of rat with about 5.3% damage of axons located at the superior temporal region. Region of affected optic nerve 1 mm posterior to the globe by light microscope showed evidence of damaged axons with axonal swelling and myelin debris. Conclusion:Sharma’s chronic intraocular hypertension model is a reproducible and effective glaucoma model, which mimics human glaucoma with chronically elevation IOP and induced RGCs loss and damage of optic nerve. Eye Science 2004;20:25-29.

New Management of Angle-closure Glaucoma by Phacoemulsification with Foldable Posterior Chamber Intraocular Lens Implantation

Objective:To investigate the management of angle-closure glaucoma by phacoemulsification with foldable posterior chamber intraocular lens(PC-IOL)implantation.Design:Retrospective,noncontrolled interventional case series.Participants:In36eyes with angle-closure glaucoma(ACG),there were18eyes with primary acute angle-closure glaucoma(PACG),14eyes with primary chronic angle-closure glaucoma(PCCG),3eyes with secondary acute angle-closure glaucoma(SACG)and 1eye with secondary chronic angle-closure glaucoma(SCCG).Intervention:Phacoemulsification with posterior chamber intraocular lens implantation.Main Outcome Measures:Postoperative visual acuity,IOP,axial anterior chamber depth.Results:After a mean postoperative follow-up time of 8.81±7.45months,intraocular pressure was reduced from a preoperative mean of 23.81±17.84mmHg to a postoperative mean of 12.54±4.73mmHg(P=0.001).Mean anterior hamber depth was1.75±0.48mm preoperatively and2.29±0.38mm postoperatively(P=0.000).Best spectacle-corrected visual acuith in 36eyes ranged from0.01to0.7(20/200to20/30)postoperatively,which was better than preoperative VA ranging from hand movement to 0.4(20/50)(P=0.000).Conclusion:Phacoemulsification with posterior chamber foldable intraocular lens implantation can be a good alternative in treating angle-closure glaucoma,Eye Science 2000;16:22-28.

Preliminary Study on in Vitro Induced Differentiation of Embryonic Stem Cells into Neurons

Purpose;To study preliminarily in vitro induced differentiation of embryonic stem cells into neurons for further investigation of an alternative for the treatment of glaumatous neuropathy.Materials and methods:Supernatant of cultured Buffalo rat liver cells (buffalorat liver cell-conditioned medium,BRL-CM)was used for culturing embryonic stem cells(ES-D3 cell line)..Morphological features of undifferentiated ES cells were studied by HE staining and electron microscopy.Based on the methods used by Bain et al,we modified the methods and used retinoic,acid(RA) as an inducer to differentiate ES-D3 cells and cytosine arabinoside(Ara-C) as inhibitor of proliferative cells.The growth of the cells was observed under phase contrast microscope.Reuslts:ES-D3 cells cultured by BRL-CM grew in aggregates and remained undifferentiated.Electromicroscopy showed large nucleus and a large amount of mitochondria in undifferentiated ES cells and many processes on the surfaces.In the first day after the adding of retinoic acid,some neuron-like cells with one,two or more processes were present.In the second day after adding RA and the first day after the plus of 10μm Ara-C,a large amount of neuron-like cells appeared,with the formation of neuron-like networks.Conclusions:Combined use of RA and Ara-C can induce ES cells to different into neuron-like cells.Our present preliminary study might provide insights into an alternative for the treatment of glaucomatous neuropathy by the transplantation of embryonic stem cells.Eye Science 2000;16:1-6.

Bifunctional Effect of Human IFN-γ on Cultured Human Fibroblasts from Tenon's Capsule

Purpose:To study the effect of human IFN-γon in vitro cultured human fibroblasts form Tenon's capsule.Materials and methods:The effect of different concentrations of human INF-γand mitomycin-C(MMC),5-fluorouracil(5-Fu）on cultured human Tenon's capsule fibroblasts(HTCF)was measured using a MTT[3-(4,5-dimethylthiazo-2-yl)]-2,5-diphenyltetrazolium bromide;Thiazolyl blue)colorimetric assay,The results were analyzed using ANOVA of the statistical package for social sciences(SPSS)9.0 version.The difference was considered to be significant if P<0.05.Results:The effects of MMC and 5-Fu on the growth of HTCF were negative,while the effects of IFN-γon the growth of HTCF were both negative(10^2-10^4units/ml in two experiments)and positive(10^6,10^5,10units/ml in two experiments).The inhibition rate of MMC ranged from5.73%to46.9%,which was similar to the inhibition rate of 5-Fu ranged from12.49%to38.92%(P=0.351),The inhibition rate of IFN-γin two experiments was smaller than MMC and 5-Fu(P<0.05).Conclusion:IFN-γhas bifunctional effect(both enhancement and inhibition)on proliferation of cultured HTCF,The antiproliferative effect of IFN-γwas weatker than MMCand 5-FU,Further study has to bd carride out to document the inhibition of scar formation of filtration bleb by IFN-γand the molecular mechanisms of its bifunctional effect on HTCF proliferation.Eye Science2000;16:43-47.

The Changes of Pattern Reversal Visual Evoked Potentials in Normal Infants

Purpose:To study pattern reversal visual evoked potential(PVEPs)and deter-mine the developmental character and mature time of visual function in normal in-fants t different months of age.Methods:PVEPs were recorded from115normal infants at3,6,9,12moths age.P1latency for different checks(1°40′，25′，6′）was analyzed and compared to those of normal adults,Changes of N1,N2latency of PVEPs were also exam-ioned.Results:P1 latency for all checks(1°40′,25′，6′）was significantly longer at 3months than at 6months of age(P<0.05),but no significant differences can be seen after 6months of age for largen(1°40′）and intermediate(25′）checks(P>0.05).P1latency for larger checks(1°40′）reached adult level after 3months of age,but not for the intermediate check(25′）,while P1latency for small check(6′）presented the character of fluctuation.Conclusion:The visual system continued to develop after birth and appeared a certain regularity,Our results showed thatP1latency for larger check(1°40′）reached adult levels after 3months of age.ButP1latency for intermediate check still has not reached adult levels after 3months of age.To deterine the age at which adult levels are finally reached,infants of 12months and older must be tested.The reason why P1latency for smaller check(6′）presented the character of fluctuation should be the temporal tuning function developing much more slowly.Eye Science1995;11:161-164.