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Retinal stem cells transplantation combined with copolymer-1 immunization reduces interferon-gamma levels in an experimental model of glaucoma

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International Journal of Ophthalmology(English edition) 2011年第6期4卷 594-598页

作者： xia Zhou xiao-bo xia Department of Ophthalmology Xiangya Hospital of Central South University Changsha 410006 Hunan Province China

AIM: To explore the effect of immunization with copolymer-1 (COP-1) and retinal stem cells (RSCs) transplantation on interferon-gamma (IFN-gamma) levels in a rat experimental glaucoma model. METHODS: An experimental glaucoma was induced by argon laser photocoagulation of the episcleral veins and limbal plexus in the right eye of rats. Immediately following glaucoma induction, rats were immunized with COP-1. RSCs were cultured and transplanted intravitreally into the eyes of glaucoma model animals 1 week post-laser treatment. Six experimental groups were used: COP-1/RSC, PBS/RSC, COP-1/PBS, PBS/PBS, glaucoma model group, and a normal control group. The concentration of IFN-gamma in aqueous humor (AH) and serum was measured by enzyme-linked immunosorbent assay (ELISA) in each of the six groups. Retinal ganglion cell (RGC) survival was assessed by quantifying apoptosis using Hoechst staining. RESULTS: Concentrations of IFN-gamma in AH and serum of rats that had undergone glaucoma induction were higher than those of non-induced control rats. The concentrations of IFN-gamma in AH and serum of the COP-1/RSCs treated group were determined to be 2371.9ng/L and 710.9ng/L, respectively, which were significantly lower than those in the other treated groups (P<0.05). In fact, IFN-gamma levels in the dual treated group were reduced to background levels. The COP-1/RSC group had lower number of apoptotic RGCs than the other three experimental groups (P<0.05). CONCLUSION: The reduced levels of IFN-gamma in AH and serum of the COP-1/RSC group may be related to synergistic effects between RSCs transplantation and COP-1 immune modulation. It is likely that the lower levels of IFN-gamma prevented RGCs glaucomatous apoptasis.

关键词： glaucoma interferon-gamma RSC transplantation COP-1 immunization

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Small interfering RNA targeting PGC-1α inhibits VEGF expression and tube formation in human retinal vascular endothelial cells

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International Journal of Ophthalmology(English edition) 2015年第5期8卷 877-883页

作者： Jian Jiang Lu Zhang xiao-bo xia Department of Ophthalmology Xiangya HospitalCentral South University

AIMTo determine whether small interfering RNA (siRNA) of PGC-1α could inhibit vascular endothelial growth factor (VEGF) expression and tube formation in human retinal vascular endothelial cells (hRVECs).METHODShRVECs transfected with peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) siRNA were incubated for 24h and then placed into a normoxic (20%, O2) or hypoxic (1%, O2) environment for another 16h. PGC-1α mRNA and protein levels were detected by real-time PCR and Western blot. VEGF mRNA and protein levels were detected by real-time PCR and ELISA. Cell proliferation was evaluated by BrdU incorporation assay. Forty-eight hours after siRNA transfection, hRVECs were planted into Matrigel-coated plates and cultured under normoxic (20%, O2) or hypoxic (1%, O2) conditions for another 48h. The tube formation of hRVECs was observed under an optical microscope and quantified by counting the number of branch points and calculating the total tube ***-1α mRNA and protein levels were significantly reduced by PGC-1α siRNA, and VEGF mRNA and protein levels also decreased significantly. The percentage of BrdU-labeled cells in siPGC-1α groups were significantly decreased compared with control siRNA groups under normoxia and hypoxia in cell proliferation assay. In the tube formation assay, PGC-1α siRNA treated cells formed significantly fewer *** PGC-1α expression can inhibit VEGF expression in hRVECs and inhibit their ability to form tubes under both normoxic and hypoxic conditions.

关键词： peroxisome proliferator-activated receptor-γ coactivator-1α vascular endothelial growth factor small interfering RNA retinal vascular endothelial cell tube formation

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Effect of pyridone agent on blood-retinal barrier in diabetic mice

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International Journal of Ophthalmology(English edition) 2017年第6期10卷 890-895页

作者： Si-Qi Xiong Hai-bo Jiang Hui-Zhuo Xu xiao-bo xia Department of Ophthalmology Xiangya Hospital Central South University Changsha 410008 Hunan Province China

AIM： To evaluate the therapeutic effect of fluorofenidone on disrupted blood-retinal barrier in the diabetic mice and uncover its underlying mechanism. METHODS： db/db mice were randomly chosen for treatment with daily doses of fluorofenidone or placebo at 5-week-old, treatment continued until mice reach 24-week- old. Then, expression of transcriptiona factor insulin gene enhancer binding protein-1 （Islet-I） and vascular endothelial growth factor （VEGF） in murine retinas were evaluated. Retinal vascular permeability was assessed by examining the level of albumin in db/db murine retinas. Furthermore, the retinal vessel tight junction was estimated by checking the level of occludin in the murine retinal tissues. RESULTS： After occurrence of diabetic retinopthy in db/ db mice, expressions of transcritpional factor Islet-1 was found to be upregulated in db/db murine retinas compared with non-diabetic controls. Similar to expression pattern of Islet-l, VEGF were also demonstrated to be increased in retinas of db/db mice, which was accompanied by increased retinal vascular leakage and decreased tight junction protein level. Systemetic administration of fluorofenidone repaired broken retinal vascular tight junction by restoring occludin expression in db/db retinal tissue. Consequently, retinal vascular premeability were indicated to be reduced by examining the transudative albumin level in diabetic retinal tissues. both Islet-1 and VEGF expression were inhibited in the retinas of db/db mice after treatment with fluorofenidone. CONCLUSION： Fluorofenidone significantly protectes retinal tight junction and reduces retinal vascular leakage. The phenomenon can be partially attributed to reducing overexpression of Islet-1 and VEGF in diabetic retinal tissues.

关键词： pyridone agent diabetic retinopathy blood-retinal barrier

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Expression of FLT4 in hypoxia-induced neovascular models in vitro and in vivo

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International Journal of Ophthalmology(English edition) 2011年第1期4卷 26-29页

作者： Jiao-Lian Liu xiao-bo xia and Hui-Zhuo Xu Department of Ophthalmology Xiangya Hospital Central South University Changsha 410008 Hunan Province China

AIM: To investigate the expression of FLT4 in retina with oxygen induced retinopathy (OIR) and in brain endothelial cell lines (bEnd3) under hypoxia conditions in mice. METHODS: Fifty-two one-week-old C57BL/6J mice were divided into control group and hypoxia group. The mice of hypoxia group were exposed to 75% oxygen for 5 days and then returned to the room air to induce retinal neovascularization. Mice in control group were raised in the environment of room air at the same time. The expressions of FLT4 mRNA and protein were checked with RT-PCR and Western Blot analysis at postnatal day 14, 17 and 21 (P14, P17 and P21) respectively. 125mmol/L CoCl(2) were added to the culture medium of bEnd3 cell, proteins were extracted in 12, 24, 48 and 72 hours and FLT4 levels were examined by Western Blot analysis. RESULTS: The mRNA and protein level of FLT4 Expressed in P14 and P17 OIR mice retina statistically up-regulated as compared with those in control group, but there was no statistical difference between OIR group and control group at P21. FLT4 levels increased significantly in 12, 24 and 48 hours hypoxia intervened bEnd3 cells, its levels in 72 hours raised mildly but showed no significance. CONCLUSION: FLT4 levels increase in OIR mice retinas and bEnd3 cells in hypoxia. It may play an important role in endothelial cells proliferation in hypoxia and retinal neovascularization in OIR mice.

关键词： FLT4 OIR mice retinal neovascularization

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Congenital fibrovascular pupillary membranes: case series with pathological correlation and surgical treatment

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International Journal of Ophthalmology(English edition) 2022年第4期15卷 669-671页

作者： Yi Zhou Cong Fan xiao-bo xia Jian Jiang Eye Center of xiangya Hospital Central South University Changsha 410008Hunan ProvinceChina

Dear Editor,We present three cases of congenital fibrovascular pupillary membranes(CFPM) with pathological correlation and surgical treatment. CFPM was first described by Cibis et al;as a new entity of anterior segmen... 详细信息

关键词： membranes congenital surgical

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Analyses on the misdiagnoses of 25 patients with unilateral optic nerve sheath meningioma

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International Journal of Ophthalmology(English edition) 2016年第9期9卷 1315-1319页

作者： Jun-Feng Mao xiao-bo xia xiang-bo Tang Xue-Yong Zhang Dan Wen Department of Ophthalmology Xiangya HospitalCentral South University

AIM：To investigate clinical features of optic nerve sheath meningioma（ONSM） that was misdiagnosed,and to find methods to reduce the ***：Retrospective series ***-five misdisgnosed patients with unilateral ONSM were collected from Jan.2008 to Jan.2015 and the clinical records ***：Patients were misdiagnosed with acute papillitis most frequently（P=17）,immediately followed by optic atrophy（P=8）,ischemic optic neuropathy（P=5）,acute retrobulbar optic neuritis（P=5）,optic disc vasculitis（P =3）.For each patient,the minimum frequency of misdiagnoses was once and the maximum was 4 *** for the lasting time of being misdiagnosed,the shortest was 1.5mo and the longest was 45 *** cases（84%） were once treated with glucocorticoids,and its side effects was found in seventeen *** patients（80%） complained with varying degree of vision *** a definite diagnosis was made,sixteen cases（64%） showed slight exophthalmos and eighteen cases（72%） had the tubular ***：ONSM without loss obvious exophthalmos is easily misdiagnosed in clinic,and for most of these ONSMs are tubular.

关键词： optic nerve sheath meningioma misdiagnosis clinical feature

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Promotion on the differentiation of retinal Müller cells into retinal ganglion cells by Brn-3b

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International Journal of Ophthalmology(English edition) 2016年第7期9卷 948-954页

作者： Zhen-Kai Wu Lan Cao Xue-Yong Zhang Wei-Tao Song xiao-bo xia Department of Ophthalmology Xiangya HospitalCentral South UniversityChangsha 410008 Hunan ProvinceChina

AIM： To investigate the role of Brn-3b in differentiation process of stem cells derived from retinal Muller cells into the ganglion cell. METHODS： The passage culture method of Muller cells from retina of newborn Sprague Dawley rats was carried out by repeated incomplete pancreatic enzyme digestion method. The cells were detected by fluorescence- activated ceil sorter （FACS）, immunohistochemistry technology and reverse transcription-polymerase chain reaction （RT-PCR） to determine the purity. The third passage of cells was induced in the serum-free dedifferentiation medium. The expression of the specific markers Ki-67 and nestin of retinal stem cells was measured by RT-PCR and Western blot. The cell proliferation of retinal stem cells was detected by 5- Ethynyl-2＇-deoxyuridine （Edu） staining. The cells were randomly divided into 5 groups as follows： group A： Brn-3bsiRNA group; group B： Brn-3b control siRNA group; group C： pGC-Brn-3b-green fluorescent protein （GFP） group; group D： pGC-GFP group; group E： control group （without any handling）. The purified Muller cells were cultured for 3-7d, then, the percentage of ganglion cells was counted by immunofluorescence staining. RESULTS： FACS demonstrated the purity of retinal Muller cells was more 97.44%. A few spherical cell spheres appeared. Immunofluorescence staining showed that stem cells within the spheres were positive for retinal stem cell-specific markers nestin （red fluorescence, 92.94%±6.48%） and Ki-67 （green fluorescence, 85.96%± 6.04% ）. Meanwhile, RT-PCR analysis showed cell spheres in the culture to have expressed a battery of transcripts characteristic of stem cells such as nestin and Ki-67, which were absent in the Muller cells. Western blot analysis further confirmed the expression of nestin and Ki-67 in the cell spheres but not in the Muller cells. Edu staining showed most of the nuclei within the cell spheres were stained red （82.80%±6.65%）, suggesting the new cell spheres had the capacity for effectiv

关键词： Mtiller cells retinal ganglion cells Brn-3b stem cells differentiation

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Therapeutic effect analysis on the treatment of congenital glaucoma through modified combined trabeculotomy-trabeculectomy

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International Journal of Ophthalmology(English edition) 2016年第2期9卷 243-248页

作者： Zhen-Kai Wu Jing Wu Qian Tan Jian Jiang Wei-Tao Song xiao-bo xia Department of Ophthalmology Xiangya Hospital Central South University Department of Ophthalmology First People's Hospital of Changde City

AIM： To evaluate the therapeutic effect and the safety of the treatment of congenital glaucoma through modified combined trabeculotomy-trabeculectomy. METHODS： The clinical data of 27 cases （altogether 42 eyes）, which included 7 cases of infants （10 eyes） and 20 cases of teenagers （32 eyes）, of congenital glaucoma undertook modified combined trabeculotomy trabeculectomy were analyzed retrospectively. The parameters evaluated included the post operation visual acuity, the anterior chamber, the filtering bleb, the intraocular pressure, the C/D ratio, visual field, the retinal nerve fiber layer changes and the complications. RESULTS： The follow-up period was 1 to 29mo, averaging 13.3 ±7.7mo. Upon the last visit after the operation, functional filtering blebs developed in all the involved eyes. The intraocular pressure was controlled under 21 mm Hg, which was decreased by 60% when compared with that before the operation, without using any medication. There were no significant changes in the post operation visual acuity and the retinal nerve fiber layer thickness before and after the operation in teenager group （P〉0.05）, and both the post operation C/D ratio and the visual field mean defect （MD） were reduced compared with those before the operation （P〈0.05）. There were no severe complications in any of the patients. CONCLUSION： The modified combined trabeculotomy- trabeculectomy can effectively reduce the intraocular pressure and control the development of glaucoma in cases of congenital glaucoma. It is a safe and effective operative method for the treatment of congenital glaucoma

关键词： congenital glaucoma trabeculotomy trabeculectomy

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Differentiation of human embryonic stem cells derived mesenchymal stem cells into corneal epithelial cells after being seeded on decellularized SMILE-derived lenticules

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International Journal of Ophthalmology(English edition) 2019年第5期12卷 717-724页

作者： Yao Chen Ye-Wei Yin Yang Zhao xiao-Ying Wu Kelly Young Wei-Tao Song xiao-bo xia Dan Wen Department of Ophthalmology Xiangya HospitalCentral South University Department of Veterans Affairs Miami VA Healthcare System

AIM: To evaluate the feasibility of mesenchymal stem cells(MSCs) to differentiate into corneal epithelial cells after being seeded on the decellularized small incision lenticule extraction(SMILE)-derived lenticules. METHODS: The fresh lenticules procured from patients undergoing SMILE for the correction of myopia were decellularized. The MSCs were subsequently cultivated on those denuded lenticules. The MSCs without lenticules were used as a control. The proliferation activity of the MSCs after seeding 24 h was quantitatively determined with the Cell Counting Kit-8(CCK-8) assay. Immunofluorescence staining and quantitative reverse transcription polymerase chain reaction(qRT-PCR) were used to assess the marker expression in differentiated MSCs. RESULTS: The data showed that both fresh and decellularized lenticules could significantly promote the proliferation of MSCs, compared to that in control(P=0.02 for fresh lenticules, P=0.001 for decellularize ones, respectively). The MSCs seeded on both lenticules were positive for cytokeratin 3(CK3) staining. The expression of CK3 increased 5-fold in MSCs seeded on fresh lenticules and 18-fold on decellularized ones, compared to that in control. There was a significant difference in the expression of CK3 in MSCs seeded on fresh and decellularized lenticules(P0.05), while the expression of these markers was decreased in MSCs seeded on decellularized ones. CONCLUSION: These results suggest that the decellularized lenticules might be more suitable for MSCs to differentiate into corneal epithelial cells, which offersthe prospect of a novel therapeutic modality of SMILEderived lenticules in regenerative corneal engineering.

关键词： mesenchymal stem cells small incision lenticule extraction lenticules decellularization corneal epithelial cells

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Treatment of myopic foveoschisis via macular buckling and vitrectomy

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International Journal of Ophthalmology(English edition) 2017年第5期10卷 815-818页

作者： Si-Qi Xiong Hai-bo Jiang Fang-Ling Li Yan-Xiu Li Jie Yang xiao-bo xia Hui-Zhuo Xu Department of Ophthalmology Xiangya Hospital Central South University Changsha 410008 Hunan Province China

The aim of the present study was to evaluate the efficacy and safety of the treatment of myopic foveoschisis patients using the macular buckling with L-shaped titanium plate and silicon sponge combined with vitrectomy. The data of the patients who underwent macular buckling combined with vitrectomy was collected. The study recorded the following parameters：best corrected visual acuity（BCVA）, axial length, intraocular pressure, central macular thickness, and the position of the titanium plate. Following the surgery, the BCVA of the included patients were improved, whereas the axial lengths were reduced followed by resolution of the foveoschisis compared with that noted prior to the operations. All patients had orbital CT examination and the results indicated that the titanium plates were appropriately placed and were not in contact with the optic nerve. Therefore, it is effective to treat myopic foveaschisis by macular buckling using the L-shaped titanium plate and silicon sponge in the presence of vitrectomy.

关键词： myopic foveoschisis macular buckling vitrectomy

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