Objective To establish a method for primary culture of iris pigment epithelial cells(IPE). MethodsEnzyme-Assisted microdissection was used to isolate and cultivate the IPE cells.An identification was made with microscopic and immunohistochemical observations.Results IPE were successfully sultured and showed on differences with RPE in primary culture and subculture.ConclusionEnzyme-Assisted microdissection is a reliable and quick method for the isolation of IPE.
Objective
To observe the morphological changes of dendrite and soma in retinal ganglion cells (RGCs) which subsisted in early diabetic rats.
Methods
The RGCs of 3-months-course diabetic rats and coeval normal rats were marked by gene gun techniques. To collect RGCs photographs by Leica microscope with Z axis and CCD camera；to observe the changes of diameter, variance of structural features in dendritic field and somata after classification which according to the size and morphology. Thy-1 antibody marks on the retinal RGCs, taking a photograph under fluorescent microscope, counting the changes of retinal RGCs density in early diabetic rat.
Results
In three-month diabetic rats,the density of retinal RGCs was decreased obviously. Morphological changes of RGCs in the dendritic fields were observed with gene gun technique. There was no severe variation in all kinds of the bole of cell dendrite, in which some only showed crispation partially and sparseness also twisting in the dendritic ramus. The mean diameter of dendritic field and soma in class A of diabetic rats was (401plusmn;86) mu;m, the mean diameter of dendritic field in control group was (315plusmn;72) mu;m,compared with each other, there is statistically significant differences (t=21.249，Plt;0.001); the mean diameter of soma in class A of diabetic rats was (24plusmn;6) mu;m, the mean diameter of soma in control group was (22plusmn;5) mu;m, compared with each other, there is no statistically significant differences (t=0.927，Pgt;0.05); the mean diameter of dendritic field and soma in class B of diabetic rats were (170plusmn;36)、(14plusmn;2) mu;m respectively, in control group were (165plusmn;36)、(16plusmn;2) mu;m, the mean diameter of dendritic field and soma in class C of diabetic group were(265plusmn;78)、(17plusmn;5) mu;m respectively, in control group were (251plusmn;57)、(17plusmn;4) mu;m , compared with each other, there are on statistically significant differences(t=1.357，0.798，0.835，1.104，Pgt;0.05).
Conclusions
In short-term diabetes, the survived RGCs show good plasticity in adult diabetic rats, especially in class A. The changes of dendrites were more sensitive than the soma, which could be the leading index of the morphologic changes of RGCs in the early stage. The good plasticity showed by the RGCs and the time window from changing in dendrite to cell death provide us many evidences not only for the research but also for the nerve protection in clinic.
(Chin J Ocul Fundus Dis,2008,24:249-254)
ObjectiveTo investigate the medium and long-term influence of silicon oil versus heavy silicone oil on rabbit retinas.
Methods28 health standard rabbits were randomly divided into A, B and C groups, with 12, 12 and 4 rabbits respectively. All rabbits received routine vitrectomy and tamponade with silicone oil (group A), or heavy silicone oil (group B) or balanced salt solution (group C). After 4, 8, 12 and 24 weeks, the retinal b-wave amplitude was measured by ERG, posterior retinal thickness was measured by optical coherence tomography (OCT). Retinal ultrastructure and tissue morphology were observed by transmission electron microscopy and optical microscopy.
ResultsCompare to group C, the b-wave amplitude decreased at 4 weeks after surgery, and decreased at 8 weeks after surgery for group B, and decreased at 8 weeks after surgery, and decreased at 24 weeks after surgery for group A. The decreases were greater in group B than group A at 8, 12, 24 weeks after surgery, the difference was statistically significant (P < 0.05). The posterior retinal thickness of group A and B was thinner than group C at 24 weeks after surgery (P < 0.05). The decreases were greater in group B than group A, the difference was statistically significant (P < 0.05). Transmission electron microscopy and optical microscopy revealed severe pathological changes of retinal ultrastructure and morphology in group A and B rabbit eyes, at 12 weeks and 8 weeks after surgery respectively. The changes were more severe in group B and group A, including edema and necrosis in cone/rod cells, in disk membranes, mitochondria, cytoplasm, nucleus and other organelles. The morphological changes were also more severe in group B and group A, including degenerations of ganglion cell layer, inner nuclear layer changes. Those changes became more severe when the tamponade time extended.
ConclusionThe heavy silicone influence on visual function, ultrastructures, histomorphology of rabbit retinas is much worse than the silicon oil, and the effect is more significant with its time prolong.
Objective
To observe the enzymic histochemical and ultrastructral changes of cryopreserved human retina.
Methods
To compare the activity of lactate dehydrogenase (LDH)， succinate dehydrogenase (SDH) and ATPase in cryopreserved retina with those in fresh retina and to observe the histological and ultrastructural changes of cryopreserved retina.
Results
There was no statistical difference between the activity of LDH，SDH and ATPase in fresh and in cryopreserved retina. Histologically, in the cryopreserved retina, fluid in neural fiber and outer plexiform layers, as well as in cone and rod layer, was sligthly more than normal. The ultrastructure is normal except that the mitochondria was swollen in different degree.
Conclusion
Cryopreservation may be an effective method for keeping the retinal cells alive for a long period and might free the transplantation from dependance on aviability of fresh dornor tissue.
(Chin J Ocul Fundus Dis，2000，16：139-212)
PURPOSE:Investigating on histopathologic changes of the photoreceptors in retinitis pigmentosa.
METHODS:Observation of the photoreceptors of retinitis pigmentosa in 11 eyes among 9 cases using light and electron microscope.
RESULTS: The pathologic changes of the photoreceptors were found to be mostly marded at the equatorial area and less at the periphery,posterior pole and macular region of the retina. In relatively early cases,degeneration and shortening of outer segments,reduction or loss of connecting cilia,stubby inner segments and swollen mitochondria Were the predominant findings. In advanced cases,the inner and outer segments and connecting cilia were diminished with reduction of nuclei in number and disarangement,cellular degeneration and disorganization. The outer limiting membrane adhered to RPE or Bruch membrane. The spaces left over by the above pathologic
changes were replaced by the displaced Muuml;ller cells and their hypertrophic processes. Also there were degeneration of the RPE cells,and some of them might migrate into the retina.
CONCLUSION:Obvious invasions of pathologic processes in photoreceptors of the retina did present in patients with retinitis pigmentosa.
（Chin J Ocul Fundus Dis,1996,12:160-162）
Objective To investingate the ultrastructural changes of retinal pigment epithelium(RPE) and its permeability in spontaneously hypertensive rats(SHR)and explore the relation between these changes and hypertensive retinopathy.MethodsThe ultrastructure of RPE cells in the SHR aged five,six,seven months wasobserved with transmission electronmicroscope and compared to its normotensive control strain(WKY) with the same age.Then,lanthanum tracer procedures were carried out to investigate pathological changes of the blood-retinal barrier.Results (1)In SHR the main pathological changes involved swelling of mitochondria,enlargement of endoplasmic reticula,decrease of RPE cell infolding,and sparseness of microvilli.These degenerations were more serious in older rats with higher blood pressure.(2)The breakdown of outer blood-retinal barrier with permeation of lanthanum tracers were evident in SHR aged six or seven month,however,in WKY and five-month SHR the traces were prevented from passing by tight junctions.ConclusionThe degeneration of RPE owing to ischemia and anoxia arises in early periosd of hypertensive retinopathy.The pathological changes of ultrastructure and permeability might interact with the damage of visual cells and play a main role in the hypertensive retinopathy.
ObjectiveTo investigate the histological characteristics of autogenous hamstring grafts after anterior cruciate ligament (ACL) reconstruction.MethodsThe patients who underwent arthroscopic single-bundle ACL reconstruction with autogenous hamstring tendons and were followed up at least 4 years and also underwent second-look arthroscopy between March 2017 and December 2017 and met the selection criteria were considered for enrollment. Graft quality under arthroscopy was evaluated as good remodeling group (GRG, the total scores were 4-6) and poor remodeling group (PRG, the total scores were 1-3) according to synovial and vascular coverage, the apparent tension of the grafts, the thickness and retear of the grafts. During the second-look arthroscopic procedures, ACL graft biopsies were performed. Normal ACL tissues harvested from the patients under 60 years old who underwent total knee arthroplasty were designated as normal controls. Graft vascularity, cellular morphology, cellular metabolism, and collagen fibril distribution were analyzed.ResultsThe 18 specimens (11 cases of GRG group and 7 cases of PRG group) and 9 native ACL biopsied tissue sample were enrolled into the study. Arthroscopy scores were 2-6 (mean, 4.7). The biology under light microscopy of GRG group was similar to that of native ACL in control group. There was no significant difference in the scores of graft vascularity and cellular morphology between GRG group and control group (P>0.05), while PRG group was significantly lower than the other two groups (P<0.05). Transmission electron microscope evaluation showed that GRG group and control group had better collagen fibril distribution and lower levels of cellular metabolism than PRG group (P<0.05). There was no significant difference in cellular metabolism between GRG and control groups (P>0.05), while collagen fibril distribution score of GRG group was significantly lower than that of control group (P<0.05).ConclusionWhile good remodeling grafts under arthroscopy in histological maturation period was proved to be more similar to normal ACL on ultrastructure properties under light and electron microscope, ultra structural differences regarding collagen fibril distribution still persist.
Objective
To study on the ultrastructural characteristic of segments of photoreceptors from neonatal retinas for supporting donor retina choice of retinal transplantation．
Methods
Photoreceptors from neonatal calf and adult calf were analysed by scanning electron microscopy and transmission electron microscopy．
Results
Segments of photoreceptors from neonatal calf appeared the mushroom pattern, in which, distal end of outer segment which was ball-shaped formed the head with mushrooms appearance, and the inner segments along with some of outer segments formed the body with mushrooms appearance． Within the outer segment, plasma membranes of adjacent evaginations form a disk subsequently． The a rray of most disks were vertical to the entire length of segments, but some were parallel and slope to．Owing to the incomplete formation, some rim of disk near distal end of outer segment revealed step-shaped appearance． The distal end of outer segment displays some processes consisted of membranous discs, much vesicular material and mitochondria, much rough endoplasmic reticulum (RER) and numerous polyso mes．Segments of photoreceptor connected with outer nuclear layer via the external limiting membrane．
Conclusion
The typical morphol ogical structures of outer segments suggest the immature and b gowth ability of photoreceptors of the retina of neonatal calf, and therefore the competence for donor material of retinal transplantation．
(Chin J Ocul Fundus Dis, 2001,227-229)
Objective:To observe the intervention effect of the tetra methylpyraz ine on the rds mice with retinitis pigmentosa.
Methods:A total of 84 rds mice were randomly divided into 2 groups, with 42 mice in each group. The mice in the experimental group underwent intraperitoneal cavity injection with hydrochlor i c tetramethylpyrazine (80 mg/kg, twice per day) at the date of birth and till 35 days after birth, whereas the normal saline was injected into the intraperito n eal cavity of rats in the control group. The mice were sacrificed 0, 3, 7, 14, 2 1, 28, 35 days after birth, and the eyeballs were enucleated for the routine pat hologic examination with the light microscope. The apoptosis of photoreceptor ce ll nuclei was detected by terminal deoxynucleotidyl transferasemediated dUTP n i ck endlabeling (TUNEL) technigue and the expression of bcl2 in retina was de tect by immunohistochemistry method.
Results:The results of li ght microscopy s howed that the layer number of retinal photoreceptor cell nuclei with tetramethy lpyrazine treatment was increased 14, 21, 28, 35 days after the treatment compar ed with that in the control group（P＜0.01）. The results of electron-micro scope suggested that tetramethylpyrazine might reduce lesions in the photoreceptor cells and the destruction of the disc member, mitochondrion，and outer limiting me mbrane in the photoreceptor outer segment in rds mice. The apoptosis of the phot oreceptor cell nuclei reduced in rds mice 3, 7, 14, 21, 28 and 35 days after the treatment compared with that in the control group （P＜0.01）. The express ion of bcl-2 in the matrix of retinal photoreceptor cell nuclei and its inner and o u ter segments increased significantly in rds mice 3，7, 14, 21, 28 and 35 days af ter the treatment （P＜0.05）.
Conclusions:Tetramethylpyra zine might reduce ret inal photoreceptor apoptosis by upregulating the expression of bcl-2 in the m at rix of retinal photoreceptor cell nuclei or its inner and outer segments in rds mice.
Objective
To study the global and histological changes of myopia and explore its pathogenic mechanism.
Methods
Chicks were reared with monocular suture of eyelid. When myopia had been confirmed by optometry, eyeballs were removed and subjected subsequently to measurement and light and electron microscopies.
Results
Three dimensions in the eyeballs of suture group were all enlarged markedly and the mean diopter was -15.00D. Under the light microscope, rod outer segment elongated and connected With PREC in suture group. With micrometer measure, cartilaginous sclera thickened and retina became thinner. Under electron microscope, rod outer segment elongated and membrane disc was intact. In the cytoplasm of RPEC, the phagosomes containing fractions of the membrane disc of outer segment were remarkably decreased.
Conclusion
Early form deprivation may affect the drop of membrane disc and cause eyeball enlargement; thus, myopia forms.
(Chin J Ocul Fundus Dis,1999,15:20-23)
