Objective:To analyze reasons of re-hospitalization and treatment methods of re-discharge in patients with penetrating keratoplasty(PKP).Methods:This was a cross-sectional study. From Jan.2017 to Dec.2021, 63 eyes of 56 patients who underwent PKP in the Second Affiliated Hospital of Guangzhou Medical University were included. Their primary diseases, direct causes of re-admission and treatment methods were analyzed.Results:Among the primary causes, 19 cases were ocular trauma (33.93%), 12 cases were corneal infection (21.43%), 9 cases were corneal leucoma (16.07%), 6 cases were corneal endothelium decompensation (10.71%), 3 cases were keratoconus (5.36%), 5 cases were corneal dystrophy (8.93%), and 2 cases were immune corneal ulcer (3.57%). Twenty-nine patients (51.79%) underwent re-hospitalization after operation. The reasons for re-hospitalization were as follows: corneal graft rejection in 10 cases (34.48%), recurrence of primary disease in 5 cases (17.24%), and infection after transplantation in 4 cases (13.79%), glaucoma in 3 cases (10.34%), chronic failure of corneal graft in 3 cases (10.34%), eyeball atrophy in 2 cases (6.90%), corneal suture removal in 1 case (3.45%), and incision leakage (hypotony) in 1 case (3.45%). The treatment methods of re-hospitalized patients were: corneal transplantation in 8 cases (27.59%), drug therapy in 14 cases (48.28%), glaucoma surgery in 2 cases (6.90%), enucleation in 1 case (3.45%), artificial vitreous implantation in 2 cases (6.90%), corneal suture in 1 case (3.45%) and corneal suture removal in 1 case (3.45%).Conclusion:The primary causes of PKP are ocular trauma and corneal infections. Rejection after corneal transplantation is the first cause of hospitalization, followed by recurrence of primary disease and postoperative infection. More than half of the re-hospitalized patients need reoperation.

Microglial cells are the main immune cells of the retina. The primary culture of the retinal microglia is critically important in investigating the cells' properties and behaviors in neurodegenerative and inflammatory retinal disease. Here, we described a modified protocol of a microglial cell culture from the neonatal rat retina. In our culture protocol, the retina was isolated from the neonatal rat eye from postnatal day 1 to day 3 and trypsinized into a single-cell suspension. The cells were seeded into a T75 flask, which was pre-coated with poly-D-lysine (PDL) and cultured with dulbecco's modified eagle medium-F12 (DMEM/F12) that contained 10% fetal bovine serum (FBS) with different concentrations. Small bright rounded cells were observed on the top of mixed glial cells on the seventh day, and attained the maximum cell number on the 14th day. Then, the isolation was performed by a shaking method and isolated cells were identified with microglia markers ionized calcium-binding adaptor molecule 1 (IBA1), transmembrane protein 119 (TMEM119), cluster of differentiation 11b (CD11b), as well as astrocyte marker glial fibrillary acidic protein (GFAP) by immunofluorescence staining. Additionally, the initial plating ratio of the mixed glial cell, culture period of isolation, procedures of the isolation, as well as the purification procedure, were optimized for our primary microglial cell culture. The morphological changes and phagocytic function were performed after lipopolysaccharide (LPS) stimulation. Moreover, the release of pro-inflammatory cytokines at different time points of LPS activation were measured. In the present study, we found that the concentration of one retina/T75 flask could harvest the largest number of microglial cells. Besides, we continuously cultured the mixed glial cells as long as one month and isolated the mixed glial cells as much as three times. In our study, we used an isolation-shaking rate of 200 rpm for 2h, which guaranteed the steady rate and resulted in high purification of the primary retinal-microglial cells, with no need of an additional purification procedure. In conclusion, we provided a high-producing protocol for the primary culture of purified rat retinal-microglial cells.

PURPOSE:Human amniotic epithelial cells (HAECs) have regenerative properties and low immunogenicity, which have enabled their use without immune rejection in regenerative medicine applications, such as wound repair, corneal surgery and burn repair. The aim of this study was to explore the potential role of HAECs in the proliferation of human corneal endothelial cells (HCEnCs) and the possible mechanism of regulation.METHODS:HAECs and HCEnCs were isolated from donated tissue samples and were cultured; the collected HAEC culture medium (HAEC-Me) was added to the human corneal endothelium medium (CEM) to establish the HAEC-CM system. HCEnCs were cultured in CEM, 20%HAEC-Me, 20% HAEC-CM, 20% HAEC-CM supplemented with a GSK-3β inhibitor TWS119 or CEM supplemented with TWS119. Then, cell proliferation, apoptosis, cell cycle progression, telomerase activity, and Wnt/β-catenin pathway-related protein levels were assessed.RESULTS:We found that the HCEnCs cultured in the 20% HAEC-CM had increased proliferative capacity, telomerase activity and β-catenin and Tcf4 expression levels, and they had a decrease in the rate of apoptosis and α-SMA expression when they were compared with the HCEnCs cultured in the 20% HAEC-Me. After GSK-3β was inhibited by TWS119, HCEnCs cultured in CEM or 20% HAEC-CM had an increased proliferative capacity, telomerase activity, β-catenin/Tcf4 expression and a decreased α-SMA expression, and they had a decreased apoptotic rate.CONCLUSIONS:These data indicate that the human amniotic epithelial cells microenvironment can promote the proliferation of human corneal endothelial cells, which may be related to regulating telomerase activity and epithelial-to-mesenchymal transition (EMT) via the Wnt/β-catenin pathway.

玻璃体积血的治疗因积血量的不同而不尽相同,少量或中等量的玻璃体积血可在3～6个月内自行吸收,而大量的玻璃体积血则需行玻璃体切除术治疗[1].不同病因导致的玻璃体大量积血,其手术方式的选择及预后有所区别,故术前明确玻璃体大量积血的病因尤为重要.但由于玻璃体严重混浊、术前无法详细检查眼底,很难判断玻璃体大量积血的病因,因此,对玻璃体大量积血的病因进行总结与分析,将有助于术前对病因的初步判断并为个性化治疗方案的制定提供依据.引起玻璃体大量积血的病因很多,可简单分为外伤性和非外伤性两大类.

Objective To evaluate the efficacy of silicone oil removal with 23-gauge transconjunctival trocar-cannulas and gas-fluid exchange. Methods The data of 39 patients received vitrectomy and silicone oil tamponade were analyzed retrospectively. Silicone oil was removed through the 23G cannula more than 2 months later. The residual oil was removed by gas-liquid exchange. The vitreous cavity was filled with air partly. Results The time for draining out the silicone oil was 3. 5 -6. 5 min ,with average of (4. 63 ±0. 98) min and the time of complete operation was 15 -35 min, with average of (20. 1 ±6. 2) min,respectively. The postoperative BCVA improved in 29 cases (74. 4％), unchanged in 5 cases (12. 8％), and experienced mild deterioration in 5 cases (12. 8％). Three cases with residual oil bubbles were observed. No vitreous hemorrhage, retinal redetachment or choroid detachment occurred during the follow-up. IOP was 6 mmHg in 1 case (1 mmHg =0. 133 kPa), and the water leakage of the puncture site was sewed up. Conclusion The surgery of silicone oil removal with 23G transconjunctival sutureless trocarcannulas and gas-fluid exchange is effective, safe and quick.

Exothermic dimethyl ether(DME)combustion can be thermally coupled with endothermic steam re-forming to decrease reactor size and increase energy efficiency.In this study,Pt-based catalysts(with different carriers,Pt loadings,and co-catalysts)were prepared,and the dimethyl ether light-off characterirstics at low temperature were investigated.The experimental results showed that the catalyst supports of HZSM-5 exhibited better light-off performance than others,and the co-catalysts of Ce can improve the catalytic activity by reducing the ignition temperature by 58℃ compared with Pt/HZSM-5 catalyst.In addition,the experiment on DME cata-lytic combustion over Pt-Ce/HZSM-5 demonstrated that higher combustion efficiency can be achieved at the excess air ratio of 1.1,and the catalyst Pt-Ce/HZSM-5 maintained good activity and stability after 100h in the stability test.

Background Researching the pathological characteristics and components of cells in internal limiting membrane of idiopathic macular hole (IMH) has an important clinical significance for the prevention of IMH.However,the study results are still disputable.Objective This study was to investigate the histopathological features of internal limiting membrane of IMH and the types of cells inside it, and explore the pathomechanism of IMH.Methods Seven specimens of internal limiting membrane were obtained during the vitrectomy with IMH patients in the Second Affiliated Hospital of Guangzhou Medical University from February 2012 to August 2013 under the informed consent of patients.The histopathological examination was performed for the structural observation and cellular identification of internal limiting membrane.The expression and location of glial fibrillary acidic protein (GFAP), CD45 and CD44 in internal limiting membrane were examined by using immunochemistry and immunofluorescence technology.Results All the seven specimens showed continuous undulating membrane with red staining.Two specimens appeared to be uniform in thickness and few cells were distributed in the specimens.The internal limiting membranes were uneven in thickness in the other specimens with retinal pigment epithelial cells,neuroglia cells,fibrocytes,macrophages and lymphocytes in them.Immunochemistry showed the positive expression of GFAP in the outer layer of the specimens.CD45 positive cells were detected in the internal limiting membranes, and CD44 was detected in the inner layer of the specimens.Conclusions Few cells exist in the internal limiting memranes of IMH.However,neuroglia cells and CD45 positive cells emerge in the internal limiting memranes of stage 3 or above IMH eyes, indicating the proliferation of cells and immuno-inflammation response exist during the IMH development.The up-regulation of CD44 expression promotes inflammatory response of internal limiting memranes.

Objective To compare the efficacy of scleral buckling surgery and vitrectomy for macula-off rhegmatogenous retinal detachment (RRD).Methods Forty-five eyes of 45 macula-off RRD patients with mild-to-moderate proliferative vitreoretinopathy (PVR) were randomly divided into two groups according to the random number chart.Twenty-seven eyes of 27 patients in group A were treated with vitrectomy and 18 eyes of 18 patients in group B were treated with scleral buckling surgery.The condition of retinal reattachment,vision,intraocular pressure (IOP),complications and subretinal fluids absorption condition were observed.Results There were not statistically significant differences in anatomic success rate and visual acuity improvement between the two groups (Fisher' s exact probability test,P =0.155;x2 =0.417,P =0.519).The incidence of postoperative complicated cataract of group A was higher than that of group B and the statistically significant difference was found (x2 =5.561,P =0.018).There were not statistically significant differences in the incidence of transient high IOP,epiretinal membrane or metamorphopsia (x2 =1.831,0.011,0.234;P =0.176,0.915,0.628).The incidence of subretinal fluids one month after the surgery of group B was higher than that of group A and the difference was statistically significant (x2 =15.162,P =O.000).Conclusion Both scleral buckling surgery and vitrectomy are effective methods for macula-off RRD patients with mild-to-moderate PVR.Though the trend is toward PPV benefited from transconjunctival sutureless vitrectomy (TSV),the scleral buckling surgery can still be considered as an effective treatment choice.

AIM:To investigate whether the abnormal differentiation of the pterygium epithelium is related to the extracellular signal-regulated kinase (ERK) signaling pathway in vitro.METHODS:The expression levels of phosphorylated ERK (P-ERK), keratin family members including K19 and K10 and the ocular master control gene Pax-6 were measured in 16 surgically excised pterygium tissues and 12 eye bank conjunctiva. In colony-forming cell assays, the differences in clone morphology and in K10, K19, P-ERK and Pax-6 expression between the head and body were investigated. When cocultured with the ERK signaling pathway inhibitor PD98059, the changes in clone morphology, colony-forming efficiency, differentiated marker K10, K19 and Pax-6 expression and P-ERK protein expression level were examined by immunoreactivity and Western blot analysis.RESULTS:The expression of K19 and Pax-6 decreased in the pterygium, especially in the head. No staining of K10 was found in the normal conjunctiva epithelium, but it was found to be expressed in the superficial cells in the head of the pterygium. Characteristic upregulation of P-ERK was observed by immunohistochemistry. The clone from the head with more differentiated cells in the center expressed more K10, and the clone from the body expressed more K19. The P-ERK protein level increased in the pterygium epithelium compared with conjunctiva and decreased when cocultured with PD98059. The same medium with the ERK inhibitor PD98059 was more effective in promoting clonal growth than conventional medium with 3T3 murine feeder layers. It was observed that the epithelium clone co-cultured with the inhibitor had decreased K10 expression and increased K19 and Pax-6 expression.CONCLUSION:We suggest ERK signaling pathway activation might play a role in the pterygium epithelium abnormal differentiation.

The CuZnAl/HZSM-5, CuZnAlCr/HZSM-5, CuZnAlZr/HZSM-5, CuZnAlCo/HZSM-5, and CuZnAlCe/HZSM-5 catalysts that were prepared by a co-precipitation method was used for hydrogen production from steam reforming of dimethyl ether (SRD) in a metal foam microreactor. These catalysts were characterized by means of XRD, TPR, SEM and BET surface areas. The results showed that promoter Cr can reduce the average pore diameter and reduction temperature of catalyst. The conversion of dimethyl ether and hydrogen yield reaches 99% and 95% respectively over CuZnAlCr/HZSM-5 catalyst under a relatively lower reaction temperature. The obtained hydrogen-riched gas is easy to purify and meet the need of polymer electrolyte membrane fuel cell. The effects of reaction temperature, space velocity and steam to DME ratio on SRD were investigated in a metal foam micro-reactor. At the conditions of T = 250 degrees C, the space velocity of 3884 ml/(g h), steam to DME = 5, DME conversion of >97% were obtained over the CuZnAlCr/HZSM-5 catalyst without obvious deactivation during 50 h. Copyright (C) 2014, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.

Background Recent studies indicated that human amnion mesenchymal stem cells (hAMSCs) can be induced to differentiate into multiple types of cells in vitro,but whether the hAMSCs can differentiate into ocular surface cells has not been reported yet.Objective This study was to investigate the feasibility of inducing differentiation of hAMSCs into ocular surface cells by co-culturing with human bulbar conjunctiva fibroblasts (hBCFs).Methods This study was approved by Ethic Committee of Affiliated Second Hospital of Guangzhou Medical University.HAMSCs were isolated from placenta under the informed consent of healthy delivery women.hAMSCs were cultured,passaged and identified by detecting the expressions of CD44,CD45,CD73,CD90 in the cells with flow cytometer,osteogenesis and adipogenic differentiation experiments.Human conjunctival tissue was obtained during the eye operation under the informed consent of patients and hBCFs were isolated and cultured with explant culture.The cells were divided into the hAMSCs culture group and the hAMSCs and hBCFs co-culture group and cultivated in Transwell chambers for 7 days.The expressions of cytokeratin 19 (CK19) and α-smooth muscle actin (α-SMA) in the cells were assayed by immnofluorescence technique.Results Cultured hAMSCs showed the slender shape and cell body enlarged with passage.CD44,CD73 and CD90 were expressed in the cells,and the expression of CD45 was absent.After 3-4 weeks of osteogenesis and adipogenic induce,the cells showed red staining for alizarin and oil red O.In the co-culture group of hAMSCs and hBCFs,hAMSCs presented the epithelioid cell-like in shape and showed the positive response for CK19 and weaker response for α-SMA.However,in the hAMSCs culture group,the cells showed the positive response for α-SMA and absent response for CK19.Conclusions The hAMSCs can differentiate into ocular surface cells after being induced by hBCFs.And the differentiation mechanism is possibly relevant to mesenchymal cells epithelium.

To enhance the heat and mass transfer during dimethyl ether (DME) steam reforming, a micro-reactor with catalyst coated on nickel foam support was designed and fabricated. A two-dimensional numerical model with SIMPLE algorithm and finite volume method was used to investigate 1) the fluid flow, 2) the heat transfer and 3) chemical reactions consist of DME hydrolysis, methanol steam reforming, methanol decomposition and water gas shift reactions. Both the numerical and the experimental results showed that the DME conversion in the micro-reactor is higher than that in the fixed bed reactor. The numerical study also showed that the velocity and the temperature distribution were more uniform in the micro-reactor. Wall temperature, porosity and steam/DME ratio have been investigated in order to optimize the process in the micro-reactor. The wall temperature of 270 °C and the steam/DME feed ratio of 5 were recommended. Meanwhile the results indicate that a larger porosity will give a higher DME conversion and CO concentration.

目的 评价Avastin(贝伐单抗)在体外对翼状胬肉成纤维细胞的增殖抑制作用以及细胞毒性.方法 通过不同浓度Avastin(0.25 g·L-1、0.50 g·L-1、1.00 g·L-1、1.50 g·L-1、2.50 g·L-1)处理人翼状胬肉成纤维细胞12h、24 h、48 h、72 h,观察成纤维细胞的增殖抑制、药物细胞毒性以及凋亡蛋白的表达. WST-1试剂检测细胞增殖,LDH试剂盒检测细胞毒性,Western blot方法检测增殖蛋白以及凋亡蛋白的表达.结果 48 h及72 h Avastin组与正常组相比光密度(0D)值差异有统计学意义(P =0.034、P =0.016).与12 h相比,0.25 g·L-1Avastin组及0.50 g·L-1Avastin组作用24 h、48 h、72 h后细胞增殖抑制率均无明显提高(均为P＞0.05),1.50 g·L-1Avastin组及2.50 g·L-1Avastin组作用48 h、72 h后细胞增殖抑制率均明显提高(均为P ＜0.05).与0.25 g·L-1Avastin组相比,0.50g·L-1Avastin组在各个时间点(除12 h外)细胞增殖抑制率均无明显提高(均为P＞0.05),1.50g·L-1Avastin组、2.50 g·L-1Avastin组在各个时间点抑制率均明显提高(均为P＜0.05).Avastin各浓度组与LDH最大释放组(加入裂解酶)对翼状胬肉成纤维细胞LDH释放率差异均有统计学意义(均为P＜0.05),各浓度组之间差异均无统计学意义(均为P＞0.05).Avastin可抑制p-CyclinD1表达,促进Caspase3的表达,与细胞对照组相比差异均具有统计学意义(均为P ＜0.05).结论 Avastin可以通过抑制p-CyclinD1以及促进Caspase3的表达抑制翼状胬肉成纤维细胞的增殖,但无细胞毒性,进一步证实了该药物的安全有效性.

Purpose: Epithelial-mesenchymal transition (EMT) of normal conjunctival tissues is a major reason of pterygium generation; alpha-smooth muscle actin (alpha-SMA) is a maker in EMT. In this study, we investigated if human amniotic mesenchymal stem cells (hAMSCs) can inhibit alpha-SMA expression and migration of human pterygium fibroblasts (hPFs) in vitro.Materials and methods: Coculture of hAMSCs and hPFs was completed by using a Transwell coculture system. The alpha-SMA expression of hPFs was detected by immunocytochemistry and western blot analysis. The migration ability of hPFs was measured by means of a migration assay. Immunoreactivity for alpha-SMA was detected in all pterygium fibroblasts examined.Results: The expression of alpha-SMA in cocultured hPFs was significantly lower than in hPFs. Similar result was demonstrated in western blot analysis. In addition, migration assay showed that hAMSCs reduce the migration of hPFs.Conclusions: These results suggested that hAMSCs have the potential to inhibit the generation and invasiveness of pterygium in vitro.

Purpose:.To evaluate the effect of Avastin on human pterygium fibroblast migration and invasiveness.Methods:.VEGF secretion was compared between human pterygium fibroblasts and conjunctival fibroblasts by measuring VEGF-A by ELISA. The influence of Avastin on HPF migration and invasiveness was observed by wound scratch and Transwell migration assays..The expression of p-ERK1 / 2 and p-FAK was analyzed by western blotting.Results:(1)VEGF was secreted in higher amounts by human pterygium fibroblasts than by conjunctival fibroblasts.(2)Avastin treatment decreased HPF migration and invasion.(3)Avastin significantly decreased the expression of p-ERK1 / 2and p-FAK in human pterygium fibroblasts.Conclusion:.Avastin can inhibit migration and invasion of HPFs by decreasing the expression of p-ERK1 / 2 and p-FAK.

Objective To investigate ocular surface in patients after Phacoemulsification with different direction of clear cornea incisions. Methods A total of 62 patients (78 eyes) were divided into two groups:one group was through a clear corneal incision at margin level,the other was through a clear corneal incision above margin.In each group,the values such as subjective dry eye symptoms (Sx),tear break-up time (tBUT),Schirmer Ⅰ test (ST-Ⅰ),and cornea incisions fluorescein staining (FL) were measured before Phacoemulsification and 1 week,1 month and 3 months after surgery. Results All the operations were achieved in the surface anesthesia.Two groups was no statistical significance in vision,age,conjunctival congestion and aqueous flare (P ＞0.05).Two groups was statistical significance in tear break-up time (tBUT),Schirmer Ⅰ test (ST-Ⅰ) (P ＜0.05),but in cornea incisions fluorescein staining (FL) had no statistical significance (P ＞0.05).The three values were obvious changes in each group after surgery 1 week compared with the preoperative.The group in a clear comeal incision above margin,at 3 months of post-operation,the fluorescein staining (FL) and tear break-up time (tBUT) was basically equal to the level of pre-operation.But the group in a clear corneal incision margin level was also less than preoperation. Conclusions Phacoemulsification through a clear corneal incision margin level is more likely to have similar dry eye symptoms than through a clear corneal incision above margin,and also those related values abnormal appear in the early time on 1 week after operation.