Müller cells are glial cells of the retina, whose major processes cross the internal and external limiting membranes of the retina, maintaining the function and metabolism of retinal photoreceptors and neurons. Their structure and function are closely related to the development of macular hole (MH). Müller cells are involved in the formation and recovery of MH from the aspect of traction and protein, and their morphology and biological function also influence the regression of MH. The current treatment modality for MH is vitrectomy combined with internal limiting membrane (ILM) peeling, in which Müller cells play a dual role after ILM peeling in different stages of MH. And its potential to re-acquire a progenitor-like state following retinal injury with the ability to proliferate and generate new neurons making it a current research hot topic, which can be a reference and inspiration for clinical treatment.
【摘要】 目的 探讨放射性核素骨显像和血清前列腺特异抗原（PSA），碱性磷酸酶（ALP），骨特异性碱性磷酸酶（BAP）测定在前列腺癌骨转移诊断中的价值。 方法 回顾性分析2006年10月-2009年10月50例前列腺癌（PCa）患者骨显像结果及PSA、ALP、BAP测定结果。 结果 50例Pca患者骨显像阳性率为70.0%。35例Pca骨转移患者分布在PSAgt;20.0 ng/mL时占97.1%，BAPgt;20.1 μg/L时占88.6%，ALPgt;130.0 μg/L时占94.3%。血清PSA、ALP、BAP水平随着放射性核素骨显像分级的增高而逐步增高，呈高度正相关。 结论 放射性核素骨显像仍然是目前诊断PCa骨转移的主要方法；PSA、ALP、BAP亦是重要的辅助诊断指标；PSAgt;20.0 ng/mL时，患者应常规行全身骨显像检查。【Abstract】 Objective To explore the clinical value of radionuclide bone scintigraphy and measurements of serum prostate-specific antigen (PSA), alkaline phosphatase (ALP) and bone-specific alkaline phosphatase (BAP) in the diagnosis of bone metastasis in prostate cancer (PCa) patients from October 2006 to October 2009. Methods The results of bone scintigraphy, serum PSA, ALP and BAP were analyzed retrospectively in 50 PCa patients. Results The positive rate of bone scintigraphy was 70.0% in 50 PCa patients. In 35 patients with PCa bone metastasis, 97.1% of them were PSAgt;20.0 ng/mL, 88.6% were BAPgt;20.1 μg/L, and 94.3% were ALPgt;130.0 μg/L. The serum levels of PSA, ALP and BAP were increased step by step along with the advancement of bone metastatic grading from M0 to M3. They were significantly positively correlated. Conclusion Radionuclide bone scintigraphy is a major method in the diagnosis of bone metastasis in PCa patients currently. PSA, ALP and BAP are also important auxiliary diagnostic markers. Patients with the level of PSAgt;20.0 ng/mL should take a routine whole-body examination of bone scintigraphy.
Objective To investigate the inhibitory effects and possible related mechanism of OTX008 [a selective inhibitor of galectin-1 (Galectin-1)] on retinal neovascularization (RNV) in mouse model of oxygen-induced retinopathy (OIR). Methods 7-day-old (P7) C57BL/6J mice were randomly (according to random number table) divided into 4 groups including normal group, OIR group, OIR-OTX008 group and OIR-phosphate buffered saline (PBS) group. To establish the OIR mouse model, mice from all groups except normal group were expose to (75±2)% oxygen for 5 days and then to room air. OIR-OTX008 group received an intravitreal injection of 1 μl (0.25 μg/μl) OTX008 at P12, OIR-PBS group received the equal volume (1 μl) of PBS injection. Mice from 4 groups were euthanized at P17, and retinas were collected for molecular biological analysis and morphological study. RNV was evaluated by counting the number of pre-retinal neovascular nuclei and the whole-mount immunofluorescent staining of mouse retina. Cyrosections of retinas were imaged via confocal microscopy to observe the enrichment of staining of Galectin-1. Protein levels of Galectin-1, Neuropilin-1 and phosphorylation of vascular endothelial growth factor receptor 2 (pVEGFR2) were determined with Western blot. Results At P17, Galectin-1 expressed higher in retinal ganglion cell layer, inner plexiform layer and inner nuclear layer from OIR group and OIR-PBS group than normal group. Galectin-1 expressed less in cryosection retinas from OIR-OTX008 group than OIR group and OIR-PBS group. The numbers of pre-retinal neovascular cell nuclei from OIR group and OIR-PBS group were obviously more than that from normal group (t=9.314,P＜0.05). The number of pre-retinal neovascular cell nuclei from OIR-OTX008 group were obviously lower than those from OIR group and OIR-PBS group (t=8.038, 7.774;P＜0.05). The RNV tufts area (t=13.250, 12.570), non-perfusion area (t=15.590, 12.430) and hypoxic area (t=9.542, 9.928) from OIR-OTX008 group were significantly smaller than those in OIR group and OIR-PBS group (P＜0.05). Protein levels of Galectin-1 (t=24.800, 23.060), Neuropilin-1 (t=4.120, 3.530) and pVEGFR2 (t=25.880, 15.480) in the OIR-OTX008 group were significantly down-regulated than those from OIR group and OIR-PBS group (P＜0.05). Conclusion Intravitreal injection of OTX008 inhibits RNV and ameliorates retinal hypoxia in mice model of OIR possibly through down-regulating Galectin-1, Neurolinpin-1 and pVEGFR2.
ObjectiveTo investigate the effect of DJ-1 encoded by Park7 gene on retinal ganglion cells (RGC) and visual function after optic nerve crush injury (ONC) in mice.MethodsThirty-seven and 116 healthy male C57BL/6J mice were randomly divided into group normal, group ONC 2d, group ONC 5d, group ONC 7d and group control, group Park7, group Park7-ONC, group ONC and group green fluorescent protein (GFP)-ONC. Group ONC 2d, group ONC 5d and group ONC 7d were sacrificed on the 2nd, 5th and 7th day after the establishment of ONC model, and the follow-up experiments were carried out. The mice in group Park7 and group Park7-ONC were injected 1 μ recombinant adeno-associated virus (rAAV) with knocking down Park7 gene into vitreous cavity, and 1 μ l rAAV with only GFP was injected into vitreous cavity of mice in group GFP- ONC, and virus transfection was observed 4 weeks after injection. The injury of ONC was perfomed at 23 days after vitreous injection in group ONC, group Park7-ONC and group GFP-ONC, and the samples were taken for follow-up experiment 5 days after modeling. The average density of RGC was observed by immunofluorescence staining, the latencies and amplitudes of a-wave, b-wave and photopic negative response (phNR) and the amplitude of oscillatory potential (OPs)were detected by full-field flash electroretinogram,and the visual acuity of mice was measured by optomotor response (OMR). The relative expression levels of DJ-1, Bax and B lymphoblastoma / leukemia-2 (Bcl-2) protein in the retina of mice in each group were detected by Western blot. One-way ANOVA was used to compare the data between groups, and t-test was used for pairwise comparison between groups.ResultsCompared with the normal group, the relative expression of DJ-1 protein in the retina of the ONC 2 d group and ONC 5 d group increased significantly, and the difference was statistically significant (t=16.610, 5.628, P＜0.01,＜0.05). Four weeks after virus transfection, strong GFP expression was seen in the RGC layer and inner plexiform layer of the retina of mice in the Park7 group. Compared with the control group, the RGC density of the retina in the ONC group decreased significantly, and the difference was statistically significant (t=16.520, P＜0.000); compared with the ONC group, the RGC density of the retina in the Park7-ONC group decreased significantly, and the difference was statistically significant (t=6.074, P＜0.01). With the increase of stimulus light intensity, the dark adaptation a wave and b wave latency of the mice in the control group gradually shortened, and the amplitude gradually increased. The stimulus light intensity was 3 cd·s/m2. There was no statistically significant difference in the dark adaptation a wave and b wave latency and amplitude of the control group, Park7 group, Park7-ONC group, ONC group, and GFP-ONC group (Incubation period: F=0.503, 2.592; P=0.734, 0.068. Amplitude: F=0.439, 1.451; P=0.779, 0.247). Compared with the control group, the Ops and PhNR amplitudes of the ONC group mice were significantly decreased (t=15.07, 12.80; P＜0.000,＜0.001). Compared with the ONC group, the Ops and PhNR amplitudes of the mice in the Park7-ONC group were significantly decreased (t=4.042, 5.062; P＜0.05,＜0.01); there was no statistically significant difference in the PhNR latency of the mice in each group (F=1.327, P=0.287). Compared with the control group, the visual acuity of the mice in the ONC group was significantly decreased, and the difference was statistically significant (t=23.020, P＜0.000); compared with the ONC group, the visual acuity of the mice in the Park7-ONC group was significantly decreased, and the difference was statistically significant (t=3.669, P＜0.05). Compared with the control group, Park7-ONC group and ONC group, the relative expression of DJ-1 protein in the mouse retina was significantly down-regulated, and the difference was statistically significant (t=47.140, 26.920; P＜0.000,＜0.000). There was no significant difference between ONC group and GFP-ONC group (t=0.739, P=0.983). Compared with the ONC group, the relative expression of Bax protein in the mouse retina of the Park7-ONC group was significantly increased, and the relative expression of Bcl-2 protein was significantly reduced. The differences were statistically significant (t=5.960, 9.710; P＜0.05,＜0.05); the relative expression ratio of Bcl-2/Bax in the Park7-ONC group was significantly lower than that in the ONC group, and the difference was statistically significant (t=13.620, P＜0.01).ConclusionThe expression of DJ-1 encoded by Park7 gene is down-regulated after Park7 gene was knocked down, which aggravates the RGC damage and the decrease of retinal electrophysiological response and visual function in ONC injury mice.
Ocular neovascularization is a pathological change in various ocular diseases such as diabetic retinopathy, retinopathy of prematurity, central retinal vein occlusion and age-related macular degeneration, which seriously affects patient's vision. β receptors are expressed in conjunctiva, corneal epithelial cells, corneal endothelial cells, extraocular muscles, trabecular meshwork, ciliary muscle, lens and retina. β adrenergic receptor antagonists bind to β receptors to exert anti-angiogenic effects by inhibiting the expression of vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1, interleukin-6 and other angiogenic cytokines; reducing macrophage-related inflammatory response; increasing the expression of anti-angiogenic factors. In the treatment of corneal neovascularization, choroidal neovascularization, and retinopathy of prematurity, it can significantly reduce the area of neovascularization and delay disease progression. Co-administration of anti-VEGF drugs can reduce the frequency of administration of anti-VEGF drugs. At effective therapeutic concentrations, β-adrenergic receptor antagonists are well tolerated; they have broader targets than anti-VEGF drugs, which offers new treatment strategies for ocular neovascularization such as corneal, choroidal and retinal neovascularization.
ObjectiveTo observe the effects of overexpression of S100A4 protein on retinal capillary cells and retinal ganglion cells (RGC) after retinal ischemia-reperfusion injury (RIRI). MethodsOne hundred healthy adult male C57BL/6 mice were randomly divided into normal control group (group C), RIRI group, adeno-associated virus (AAV2)-S100A4 green fluorescent protein (GFP) intravitreal injection group (group S), RIRI+AAV2-GFP intravitreal injection group (group GIR), and RIRI+AAV2-S100A4-GFP intravitreal injection group (group SIR), with 20 mice in each group. The RIRI model was established using the high intraocular pressure anterior chamber method in the RIRI, GIR and SIR groups of mice. Eyes were enucleated 3 days after modelling by over anaesthesia. The number of retinal capillary endothelial cells and pericytes in the retinal capillaries of mice in each group was observed by retinal trypsinised sections and hematoxylin-eosin and periodic acid-Schiff staining; immunofluorescence staining was used to observe endothelial cell, pericyte coverage and RGC survival; The relative expression of Toll-like receptor 4 (TLR4), p38 MAPK and nuclear factor erythroid 2-related factor 2 (NRF2) in retinal tissues was measured by Western blot. One-way analysis of variance was used to compare data between groups. ResultsThree days after modeling, the endothelial cell to pericyte ratio in group C was compared with group S and SIR, and the difference was not statistically significant (F=106.30, P＞0.05); the SIR group was compared with group RIRI and GIR, and the difference was statistically significant (F=106.30, P＜0.000 1). Comparison of endothelial cell coverage in each group, the difference was not statistically significant (F=3.44, P＞0.05); compared with the pericyte coverage in group C, the RIRI group and the GIR group were significantly lower, and the difference was statistically significant (F=62.69, P＜0.001). Compared with the RGC survival rate in group C, it was significantly lower in RIRI and GIR groups, and the difference was statistically significant (F=171.60, P＜0.000 1); compared with RIRI and GIR groups, the RGC survival rate in SIR group was significantly higher, and the difference was statistically significant (F=171.60, P＜0.000 1). The relative expression levels of TLR4, p38 and NRF2 proteins were statistically significant among all groups (F=42.65, 20.78, 11.55; P＜0.05). ConclusionsPericytes are more sensitive to ischemia than endothelial cells after retinal RIRI in mice, and early vascular cell loss is dominated by pericytes rather than endothelial cells. The overexpression of S100A4 protein protects against loss of pericytes and RGC after RIRI by inhibiting the TLR4/p38/NRF2 signaling pathway.
ObjectiveTo observe the changes of macular vascular density of hypertensive patients without obvious hypertensive retinopathy (HRP).MethodsTwenty-three patients (hypertension group) diagnosed as grade 2 or grade 3 essential hypertension in Cardiology Department of Renmin Hospital of Wuhan University from January to April 2019 were enrolled in the study. Among them, there were 13 males and 10 females. The mean age was 61.6 ± 5.6 years, and the mean BCVA was 0.74 ± 0.16. The course of hypertension was more than 7 years; Keith Wagener (K-W) grade was 0 or 1. Fifteen age-matched people without hypertension were selected as the control group, among which included 8 males and 7 females. Their average age was 59.7 ± 4.4 years and the average BCVA was 0.79 ± 0.17, the K-W grades were 0. There was no significant difference (P=0.265, 1.000, 0.563) between the two groups in age (t=1.739), sex ratio (χ2=0.036) or BCVA (t=0.585). All subjects were examined by BCVA, fundus photography and OCTA. OCTA scanned the macular area in the range of 3 mm × 3 mm. The software automatically divided the image into two concentric circles with the macular fovea as the center, which are the inner ring with a diameter of 1 mm (foveal area) and the outer ring with a diameter of 1-3 mm. The blood flow density of the whole, temporal, upper, nasal and lower capillary layers within 3 mm of the macular area, and foveal avascular zone (FAZ) area, central foveal retinal thickness (CFT) were measured.ResultsSignificant differences were observed in the vascular densities of total, temporal, nasal and inferior area of maculas (t=2.188, 2.472, 5.105, 2.734; P=0.037, 0.020, 0.000, 0.010) between the two groups, while no significant differences were evidenced in foveal vascular densities and superior vascular densities (t=0.575, 0.140; P=0.570, 0.889). There was no significant difference in FAZ area or CFT between the two groups (t=0.367, 0.753; P=0.714, 0.457). Macular arches were intact in all hypertension patients.ConclusionsThe vascular densities of total, temporal, nasal and inferior area of maculas in the hypertension patients without HRP decreased. The area of FAZ did not expand, and the structures of macular arch ring were normal.
ObjectiveTo investigate the effects of Krüppel-like factor 7 (KLF7) on the survival of retinal ganglion cells (RGCs) and electroretinogram (ERG) after retinal ischemia-reperfusion (RIR) injury in mice.MethodsA total of 126 male C57BL/6J mice were randomly divided into normal group, RIR group, normal-KLF7 group, normal-green fluorescent protein (GFP) group, RIR-KLF7 group and RIR-GFP group. At the age of 8 weeks, mice of normal-KLF7 group and RIR-KLF7 group were intravitreally injected 1ul of 1.0×1012 vg/ml adeno-associated virus overexpressing KLF7 (AAV2-KLF7-GFP). Mice of normal-GFP group and RIR-GFP group were injected adeno-associated virus of AAV2-GFP with the same titer. At the age of 11 weeks, RIR injury was induced in mice of RIR group, RIR-KLF7 group and RIR-GFP group, and intraocular pressure was measured. Retinal cryosections were used to access the efficacy of virus transfection 4 weeks after AAV2-KLF7-GFP transfer. 7 days after RIR injury, RGCs’ survival rate was observed and quantified by immunofluorescent staining. ERG was performed to observe the differences in amplitudes and incubation period of scotopic ERG a-, b-wave, oscillatory potentials (Ops), photopic negative responses (PhNR). Optomotor response was performed to observe the differences of visual acuity. Expression of KLF7 was detected by western blot 4 weeks after AAV2-KLF7-GFP transfer.ResultsCompared with normal group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR group were decreased, and the differences were statistically significant (t=12.860, 7.157, 5.735, 8.953, 4.744, 9.887; P＜0.05). With the increase of light intensity, the amplitudes of scotopic ERG a- and b-wave were gradually increased while the incubation period was gradually shortened. Compared with RIR group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR-KLF7 group were increased, and the differences were statistically significant (t=6.350, 3.253, 3.695, 5.825, 5.325, 4.591; P＜0.05). Protein level of KLF7 was up-regulated in normal-KLF7 group than those in normal group, and the difference was statistically significant (t=4.105, P＜0.01).ConclusionOverexpression of KLF7 can improve RGCs’ survival rates and preserve the electrophysiological function.
ObjectiveTo compare the efficacy of internal limiting membrane (ILM) flip coverage with ILM multilayer tamponade in the treatment of highly myopic macular hole-associated retinal detachment (MHRD). MethodsA retrospective clinical study. From November 2019 to June 2022, 53 cases and 53 eyes of MHRD patients who were examined and diagnosed at the Eye Centre of Renmin Hospital of Wuhan University were included in the study. Among them, 21 cases and 21 eyes were male and 32 cases and 32 eyes were female. The age was (55.28±11.40) years. The patients were categorized into two groups: the ILM coverage group (from November 2019 to September 2020) and the ILM multilayer tamponade group (from October 2020 to June 2022) based on their surgical procedures. The ILM coverage group comprised of 11 cases involving 11 eyes, while the ILM multilayer tamponade group comprised of 42 cases involving 42 eyes. Best-corrected visual acuity (BCVA) and optical coherence tomography were conducted. BCVA was measured using standardized international visual acuity charts and transformed to logarithmic minimum angle of resolution (logMAR) visual acuity for statistical analysis. The affected eyes were all treated with standard transciliary flattening three-channel 23-gauge vitrectomy. The inverted ILM flap technique was combined with flap coverage in the inverted group, while the ILM multilayer tamponade group used circular ILM stripping to preserve the ILM in the macular area and ILM flap around the macular hole with multilayer ILM tamponade. Postoperative follow-up was carried out for a minimum of 6 months. Relevant examinations were conducted during the follow-up using the same equipment and methods as those used before surgery. The BCVA, as well as the closure of macular hole, resurfacing of the retina, and development of macular hyperplasia, were observed. ResultsIn the ILM-covered group, the macular hole was closed in 7 out of 11 eyes after 1 week of surgery. At 1 month after surgery, the macular hole was closed in all treated eyes. At 6 months after surgery, the macular hole was closed in 9 eyes, while 2 eyes were reopened. In 42 eyes from the ILM-multilayer tamponade group, the macular hole closed after surgery in 41 eyes. At 6 months postoperatively, best corrected visual acuity (BCVA) of eyes in ILM-covered and ILM-multilayer tamponade groups was 0.91±0.29 and 1.05±0.39, respectively, with no statistically significant difference between the two groups (t=1.140, P=0.260). The BCVA of the eyes in both groups showed a significant improvement compared to the preoperative period with a statistically significant difference (t=8.490, 13.840; P＜0.000 1); 6 months after surgery, 10 out of 11 eyes in the ILM coverage group had a restored retina with no detectable macular hyperplasia; 42 eyes in the ILM multilayer tamponade group had a restored retina, but 19 of these eyes had detectable macular hyperplasia. ConclusionsEither ILM flap coverage or ILM multilayer tamponade contributes to high myopic MHRD closure and improved visual acuity. Compared to ILM flap coverage, ILM multilayer tamponade results in higher and earlier rates of macular hole closure and lower rates of macular hole reopening. However, ILM multilayer tamponade may lead to a higher proportion of macular hyperplasia formation without affecting visual acuity recovery at 6 months after surgery.