Cilia are hair-like protuberance on cells of the human body that play a vital role in organs generation and maintenance. Abnormalities of ciliary structure and function affect almost every system of the body, such as the brain, eyes, liver, kidney, bone, reproductive system and so on. Retinal photoreceptor cells are one of sensory neurons which convert light stimuli into neurological responses. This process, called phototransduction, takes place in the outer segments (OS) of rod and cone photoreceptors. OS are specialized sensory cilia, and disruptions in cilia genes, which are causative in a growing number of non-syndromic retinal dystrophies, such as retinitis pigmentosa, Leber’s congenital amaurosis. These syndromes are genetically heterogeneous, involving mutations in a large number of genes. They show considerable clinical and genetic overlap. At present, there are few researches on retinal ciliopathies and clinical treatment strategy. This review shows a comprehensive overview of ciliary dysfunction and visual development related diseases, which contributes to understand the characteristics of these diseases and take early intervention in clinic.
Objective To investigate the cellular phenotype involved in experimental autoimmune uveoretinitis (EAU) and apoptosis of infiltrating cells in this inflammation． Methods Immunohistochemical staining and in situ apoptosis staining were performed using monoclonal antibodies to monocytes and macrophages (EDI),MHC calss -II antigen (OX6),T lymphocytes (R73) and TACS 1 Klenow kit on both ocular sections and wholemounts of 16 Lewis rats after immunization with interphotoreceptor retinod-binding protein(IRBP). Results EAU was induced in 12 of 16 Lewis rats with a clinical inflammation score being 1.29plusmn;0 .7.Influx of monocytes,lymphocytes and MHC class II+ cells into the uvea and retina was noted after immunization with IRBP.Apoptosis of infiltrating cells was observed in the uvea and retina and more apoptotic cells were present in the iris and ciliary body compared with the choroid and retina. Conclusion A number of cells including monocytes,macrophages,lymphocytes and MHC class II+ cells are involved in EAU induced by IRBP.Apoptosis of infiltrating cells occurs at early stage of EAU,which may greatly contribute to the rapid regression of the inflammation induced by IRBP. (Chin J Ocul Fundus Dis，2000，16：1-70)
Objective To investigate the expression and significance of inducible co-stimulator (ICOS) in experimental autoimmune uveoretinitis (EAU). Methods EAU was induced in 24 Lewis rats (immune group) by immunization with retinal S-antigen (50 mu;g) and complete Freundprime;s adjuvant, and another 4 rats were in the control group. Anterior segment of the ratsprime; eyes were observed by split microscope every day. Immunohistochemical staining was performed using polyclonal antibodies to ICOS on the sections of the spleen which were obtained from the rats in immune group at the 7th, 12th, 15th and 21st days after immunisation respectively. Western blotting was performed to investigate the dynamic expression of ICOS protein in the spleen. The same procedures were made at the corresponding time points in the rats in control group. Results A few ICOS positive cells were observed in the normal spleen. The number of ICOS positive cells in immune group increased obviously at the 7th and 12th days after immunization, reached the peak at the 15th day, and decreased at the 21st day which was still higher than that in the control group. The result of Western blotting showed that the dynamic changes of ICOS protein was identical with the changes of positive-cell number detected by immunohistochemistry. Conclusions The enhanced expression of ICOS happens before EAU occurs, which increases when the inflammation occurs and deteriorates, and decreases at the alleviative stage of EAU. It suggests that ICOS participates in the formation, development and disappearance of EAU and plays an important role in the incidence of EAU. (Chin J Ocul Fundus Dis, 2005,21:114-117)
Objective To observe the expression and investigate the significance of suppressor of cytokine signaling (SOCS) in peripheral blood mononuclear cells (PBMC) of experimental autoimmune uveitis (EAU). Methods 100 Lewis rats were immunized with interphotoreceptor retinoid-binding protein (IRBP) to induce EAU animal model, and they were divided into control group and treatment group randomly. The treatment group was administered cyclosporine A 20mg/（kgmiddot;d）after 1 to 28 days of immunization; the control group received saline buffer at equal quantity. All eyes were evaluated by slit-lamp microscopy before and after 7, 14, 21, 28 days of immunization; IL-4，IL-12，IFN-gamma; in the serum were measured by enzyme linked immunosorbent assay(ELISA); the SOCS mRNA and protein level in PBMC were measured by quantitative polymerase chain reaction (q-PCR) and western blot. Results The inflammation was most obvious at 14 days after immunization. The control group showed obvious iridocyclitis; the treatment group showed mild anterior chamber inflammation but no posterior synechia and hypopyon. The highest level of IL-12 and IFN-gamma; were observed at 14 days after immunization, followed by decline to the baseline at 28 days after immunization in control group; the highest level of IL-12 and IFN-gamma; were found at 14 days after immunization in treatment group, but the level was lower than control group obviously. Compared with the level before immunization, there are no differences at other time-point. The concentration of IL-4 decreased indistinctly in control group but increased in treatment group. SOCS1、Both of SOCS1 and SOCS5 increased to the highest level at 14 days after immunization, as 4.05 and 383 times of preimmunization in control group respectively, as 1.15 and 1.16 times in treatment group respectively. The CIS and SOCS3 mRNA increased lightly in two groups and treatment group milder than control group. Marked increased expression of SOCS1 and SOCS5 protein was detected at 7, 14, 21days than preimmunization, both of CIS and SOCS3 protein were significantly increased on 14, 21 days in control group; only SOCS1 protein was significantly increased on 14 days in treatment group and there are no differences at other time-point compared to pre-immunization. Conclusion Up-regulation of SOCS1 and SOCS5 expression maybe related to intensive response of Th1 in the development of EAU. Mild up-regulation of CIS and SOCS3 maybe associated with intensive response of Th2 which against the reaction of Th1 to carry out the dynamic immune balance.
Objective To observe the effect of Minocycline on RP process of retinal pigmentary degeneration rd mice［C3H/HeN (Pde6brd-/rd-)］. Methods 40 rd mice were divided into ten groups randomly: 5 experimental groups and 5 control groups, 4 rd mice in each group. The experimental group received intraperitoneal injection of minocycline 22.5mg/kg while the control group received saline 10ml/kg every day from the postnatal day 1 (P1) . Mice were sacrificed at P1, P7, P14, P21 and P28 respectively. Eyeballs were enucleated to carry out histology observation and apoptosis cell detection. Meanwhile, to statistically analyze the number of retinal photoreceptor cells,the thickness of outer nuclear layer (ONL)and the number of apoptosis cells. Results （1）Photoreceptor cell began to apoptosis on P7, peaked on P14, and totally disappeared on P28. （2）No statistically significant differences were found of the number of photoreceptor cells and the thickness of ONL on P7 between the experimental group and the control group. (3) The number of photoreceptor cells and the thickness of ONL in the experimental were more than that in the control group at P14, P21, P28 respectively, the differences are statistically significant（Plt;0.05）. (4) The apoptotic cells on ONL were less in the experimental group than that in the control group on P7 and P14 respectively, the difference are statistically significant（Plt;0.05）. Conclusions Minocycline appears to protect photoreceptor cell from apoptosis in the early stage of the retinal degeneration mice, but it may not completely prevent RP from occurrence.
Objective To observe the efficacy of the anti-tumor necrosis factor-alpha; monoclonal antibody (TNF-alpha; MCAb) in the treatment of experimental autoimmune uveoretinitis (EAU). Methods EAU animal models were induced by interphotoreceptor retinoid-binding protein (IRBP) R16 peptide with immunization. The rats were divided into 2 groups according to the injection times. TNF-alpha; MCAb was administered intravenously on day 6 or 4, 6 and 8 post-immunization respectively, and then to observe the clinical expression by slit-lamp microscope. Meanwhile, take the rats which did not accept TNF-alpha; MCAb as control group. Delayed type hypersensitivity (DTH) responses were measured on day 13 post-immunization of IRBP R16; the rats were killed on day 14 post-immunization of IRBP R16, and then enucleated the eyes for histopathological examination. To detect the cytokine level of IFN-gamma;, IL-4 in serum and IFN-gamma; in aqueous humor by enzyme-liked immunosorbent assay (ELISA) on day 14 post-injection. The hyperplasia responses of antigen specific lymphocyte of draining lymph node cells were detected. Results The TNF-alpha; MCAb group had mitigated ocular inflammation and decreased pathological grades compared with the control group; the IFN-gamma; concentrations in aqueous humor and serum were decreased, IL-4 was increased in serum; DTH responses were decreased; the hyperplasia responses of draining lymphocytes to IRBP R16 peptide were decreased, all the differences were statistically significant (P＜0.01). The rats accepted TNF-alpha; MCAb thrice had much better curative effect than the rats injected once (P＜0.05). Conclusions Injection of TNF-alpha; MCAb can inhibit ocular inflammation and specific immune cells of EAU remarkably and change the Th1/Th2 balance. Many times injections of TNF-alpha; MCAb were more effective than once.
Objective To detect and analyse the mutations in rhodopsin gene of members in a family affected by autosomal dominant retinitis pigmentosa (ADRP). Methods Using the polymerase chain reaction (PCR), we amplified exon 1-5 of rhodopsin gene in patients with ADRP,and analyzed it with direct sequence measuement. Results The Gly-182-Asp mutation in the rhodopsin gene was detected in most of affected members of this ADRP family, but no mutation was detected in two affected members and the control ones. Conclusion We cannot regard the Gly-182-Asp mutation in the rhodopsin gene as the pathagenic factor of the ADRP family. It is likely there is a new gene next to the rhodopsin gene. (Chin J Ocul Fundus Dis, 2002, 18: 256-258)
Objective To observe the expression of cyclin dependent kinase 5 (Cdk5) and p25 in the pathogenesis of retinitis pigmentosa (RP) in Royal College of Surgeon (RCS) rats and its relationships with apoptosis. To explore the mechanism of Cdk5 and p25 induced photoreceptor apoptosis in the pathogenesis of RP. Methods Retinas of RCS and RCS-rdy+ rats were obtained at the ages of postnatal day 17, 25, 35, 60. The retinal structure and thickness of outer nuclear layer were measured by optical microscopy. The expression of Cdk5, p25, cleave-caspase 3 in the retina was evaluated by immunohistochemistry. The protein expression of cleave-caspase 3 in the retina was determined by Western blot. The apoptosis of retinal cells was examined by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The mean absorbance value of apoptotic cells was analyzed by SPSS 17.0 software. Results The retinal thickness of the RCS rats was significantly reduced in comparison to the RCS-rdy+ rats as the postnatal days progressed, particularly in the layer of rods and cones and the outer nuclear layer. The expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increased from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above indexes in RCS-rdy+ rats. The protein expression of cleave-caspase 3 in the RCS rats was significantly increased with progression of postnatal days to postnatal 35; but there was no obvious similar change in RCS-rdy+ rats. The results of TUNEL showed that the apoptotic cells significantly increased in the outer nuclear layer of RCS rats from postnatal 17 days to postnatal 35 days, while decreased on postnatal 60 days; but there was no obvious change of above index in RCS-rdy+ rats. This study showed that there were significant correlations between the following variables: Cdk5 expression and p25 expression, Cdk5 expression and cleave-caspase 3 expression, Cdk5 expression and apoptotic cells, p25 expression and cleave-caspase 3 expression, p25 expression and apoptotic cells, cleave-caspase 3 expression and apoptotic cells. The partial correlation coefficients were 0.949, 0.808, 0.959, 0.887, 0.979, 0.852, respectively and the P value was 0.000. Conclusions The apoptotic cells significantly increases and the expression level of Cdk5, p25, cleave-caspase 3 of RCS rats increases from postnatal 17 days to postnatal 35 days. The tendency of apoptotic cells to increase is consistent with the change of Cdk5, p25, cleave-caspase 3 expression. The apoptosis of photoreceptor cells is related to increasing expression of Cdk5 and p25 in RCS rats. Cdk5 may be involved in the development of RP in RCS rats.
Objective To evaluate the therapeutic effects of vitrectomy on chronic and refractory Behcet′s diseases. Methods The clinical data of 8 patients (10 eyes) with Behcet′s diseases from 0.5 to 3 years after vitrectomy were retrospectively analyzed. Pre-and post-operative visual acuity, control of inflammation after the surgery, and the recurrence were observed and analyzed. Both of the eyes were involved in all of the patients, including complete Behcet′s diseases in 5 and incomplete in 3. Results Vitreous opacity and liquefaction were found in all of the involved eyes, dark retina and thin retinal vessels were seen in most of the eyes, and vascular obstruction or sheath-like changes in different degrees were detected in some eyes. Improvement of visual acuity was observed in all of the eyes which had undergone the treatment after the operation. There was significant difference between the visual acuity before the surgery and at the 1st and 2nd week and the 1st and 6th month after the surgery. During the follow-up period, recurrence of the inflammation was found in 3 eyes within 1 month and in 6 eyes within 6 months; no recurrence was found in 4 eyes after the operation. No acute recurrence of inflammation was found after operation in the involved eyes. Conclusion Vitrectomy for chronic and refractory Behcet′s diseases may improve the visual acuity of the involved eyes, and the surgery is safe and effective. (Chin J Ocul Fundus Dis, 2005, 21: 357-359)
Objective To observe the clinical characteristics of idiopathic retinal vasculitis, aneurysms, and neuroretinitis (IRVAN) syndrome. Methods The clinical data of 3 patients with IRVAN syndrome which were diagnosed by systemic examination, fundus photography and fundus fluorescein angiography (FFA) were retrospectively analyzed. Results Idiopathic retinal vasculitis, which was induced by retinal arterial inflammation, multiple macroaneurysms of optic disc and retinal vessels, edema of optic disc, and exudation around the optic disc, was found in all of the 3 patients, multiple arteriolar aneurysms of optic disc and retinal vascular and exudative neuroretinitis. Two patients had peripheral retinal vascular nonperfusion area, which belonged to typical IRVAN syndrome. Conclusions The clinical characteristics of IRVAN syndrome include idiopathic retinal vasculitis which only involved in artery, multiple retinal macroaneurysms which located on the dissepiment of optic disc and retinal artery, and the neuroretinitis induced by exudation of retina and optic disc because of vasculitis and aneurysms. (Chin J Ocul Fundus Dis, 2007, 23: 180-183)