Gender-specific lncRNA-miRNA-mRNA regulatory network to reveal potential genes for primary open-angle glaucoma.

BACKGROUND: Investigation of biomarkers may facilitate understanding the mechanisms of primary open-angle glaucoma (POAG) and developing therapeutic targets. This study aimed to identify potential genes based on competing endogenous RNA (ceRNA) network for POAG. METHODS: Based on long noncoding RNAs (lncRNAs), microRNAs (miRNAs) and messenger RNAs (mRNAs) from the Gene Expression Omnibus (GEO) database, we identified differential expressed lncRNAs (DELs), differential expressed miRNAs (DEMis) and differential expressed mRNAs (DEMs) and then constructed a ceRNA network. Through weighted gene co-expression network analysis (WGCNA), we identified gender-specific genes for gender-associated ceRNA network construction, followed by the protein-protein interaction (PPI) network and functional enrichment analysis to screen hub genes and reveal their functions. The expression levels of hub genes were measured in steroid-induced ocular hypertension (SIOH) mice. RESULTS: A total of 175 DELs, 727 DEMs and 45 DEMis were screened between control and POAG samples. Seven modules were identified through WGCNA and one module was associated with gender of POAG patients. We discovered 41 gender-specific genes for gender-associated ceRNA construction and then identified 8 genes (NAV3, C1QB, RXRB, P2RY4, ADAM15, VAV3, ZNF207 and TOP1), which were enriched in cell cycle-related pathways and immune-related pathways. C1QB, RXRB, Top1 and ZNF207 were highly interacted with other proteins. The expression levels of NAV3 and C1QB were downregulated in SIOH, while the levels of RXRB, P2RY4, ADAM15, VAV3, ZNF207 and TOP1 were upregulated in SIOH. CONCLUSION: This study identifies hub genes associated with the pathogenesis of gender-specific POAG and provides potential biomarkers for POAG.

Chronic hypotony management using endoscopy-assisted vitrectomy after severe ocular trauma or vitrectomy.

AIM: To report outcomes of endoscopy-assisted vitrectomy (EAV) in patients with chronic hypotony following severe ocular trauma or vitrectomy. METHODS: This was a retrospective, noncomparative case series. Ciliary bodies were evaluated using ultrasound biomicroscopy pre-operatively and direct visualisation intraoperatively. All selected individuals (seven patients/seven eyes) underwent EAV. Removal of ciliary membrane and traction, gas/silicone oil tamponade (GT/SOT), and scleral buckling (SB) were performed in selected eyes. Outcome measurements mainly included intraocular pressure (IOP) and best-corrected visual acuity (BCVA). RESULTS: Seven eyes from 7 male aphakic patients with a mean age of 45 (range, 20-68)y were included in this study; the average follow-up time was 12 (9-15)mo. GT was performed in 2 eyes; membrane peeling (MP) and SOT in 2 eyes; and MP, SOT, and SB in 3 eyes. The mean pre- and post-operative IOP were 4.5 (range, 4.0±0.11 to 4.8±0.2) mm Hg and 9.9 (range, 5.6±0.17 to 12.1±0.2) mm Hg at 52wk (12mo), respectively. BCVA improved in six eyes; one eye still showed light perception, and no bulbi phthisis was observed. CONCLUSION: Endoscopy offers improved judgment and recognition and has an improved prognosis for chronic hypotony. Therefore, endoscopy can be an effective and promising operative technique for chronic traumatic hypotony management.

Preparation of amidated pectins through enzymatic method: Structures, hydrogel properties and its application potential in fat substitutes.

In this paper, α-alanine and ß-alanine were used as modifiers to prepare α-alanine amidated pectin (α-AP) and ß-alanine amidated pectin (ß-AP) through enzymatic method. The effects of alanine and its isomer on the amidation degree and physicochemical properties of pectin were studied. Fourier transform infrared spectrum, proton nuclear magnetic resonance analysis, X-ray photoelectron spectrum and elemental analysis indicated that the amino groups from α-C and ß-C could be aminated with the carboxyl group of pectin to form the corresponding aminated pectin. The alanine grafting ratio of α-AP and ß-AP were 21.99% and 18.92%, respectively. The results showed that the dynamic viscosity of ß-AP was significantly higher than that of α-AP due to the influence of amino acid side chain. However, due to the higher alanine grafting ratio of α-AP, the strength of hydrogel prepared by α-AP was higher than that of ß-AP, and it also has the highest elastic modulus and swelling capacity. In addition, pectin, as a carbohydrate-based fat substitute, has been widely used in the field of food to simulate the smooth and delicate taste of fat. Compared with High methoxy pectin (HMP) and ß-AP, α-AP had better colloid stability and smaller hydrogel particles in the composite system. The results provide references for the application of amidated pectin in low-fat food.

Fabrication of phenylalanine amidated pectin using ultra-low temperature enzymatic method and its hydrogel properties in drug sustained release application.

In this study, pectin was modified with phenylalanine by ultra-low temperature enzymatic method to improve its gel properties. The grafting ratio of phenylalanine amidated pectin was studied under different reaction conditions. The highest value (29.21 %) was reached a reaction temperature of -5 °C and time of 12 h. Further analysis indicated that phenylalanine and high methoxyl pectin combined at the solid-liquid two phase interface under the catalysis of papain to form phenylalanine amidated pectin. Moreover, the physicochemical properties of pectin hydrogel and its feasibility as a sustained-release drug carrier were discussed. The results showed that phenylalanine amidated pectin can form hydrogel with a certain strength under acidic conditions, and there is no need to add a lot of soluble solids and divalent cations. Besides, the phenylalanine amidated pectin hydrogel as a sustained release carrier of drugs showed more sustained and complete drug release.

Utilization of the antagonistic yeast, Wickerhamomyces anomalus, combined with UV-C to manage postharvest rot of potato tubers caused by Alternaria tenuissima.

Postharvest rot of potato tubers caused by fungal pathogens is the main cause of significant economic losses, while also raising potential food safety issues. Integrated disease management, utilizing bio-safe and eco-friendly methods, represents a sustainable strategy for reducing postharvest losses in crops, including potato. In the current study, the application of the antagonistic yeast, Wickerhamomyces anomalus, combined with a UV-C treatment was evaluated for the management of postharvest Alternaria rot of potato tubers, caused by Alternaria tenuissima. Both W. anomalus and UV-C as individual treatments reduced the size of A. tenuissima infections on potato tubers, relative to the control, while the combined treatment of W. anomalus and UV-C exhibited the highest level of inhibition. W. anomalus or UV-C alone, and especially when used in combination, induced the expression of defense-related genes, including polyphenol oxidase, peroxidase, and ß-1,3-glucanase, and also increased the level of flavonoids and lignin in potato tubers. Our findings indicate that the mechanism of action by which UV-C enhances the biocontrol effect of W. anomalus against postharvest Alternaria rot includes the activation of defense-related response in potato tubers. The integration of biocontrol agents and physical treatments (e.g., UV-C) represents an effective, eco-friendly hurdle technology for managing postharvest rot in potato.

Associations of Urinary Phytoestrogen Concentrations with Nonalcoholic Fatty Liver Disease among Adults.

Phytoestrogens can alleviate some pathological processes related to nonalcoholic fatty liver disease (NAFLD). However, there are limited and contradictory studies on the relationships between phytoestrogens (especially single phytoestrogen) and NAFLD. The purpose of this study was to explore the relationships between urinary phytoestrogen concentrations and NAFLD in American adults. This cross-sectional study used the data of the National Health and Nutrition Examination Survey from 1999 to 2010, and 2294 adults were finally enrolled in this study. The concentrations of phytoestrogens were measured in urine samples, and urinary phytoestrogens were divided into tertiles according to the concentration distributions. The diagnosis of NAFLD was determined by the United States fatty liver index. The main analysis used a multivariate logistic regression model. The fully adjusted models included gender, age, race, education, marriage, poverty, body mass index, waist circumference, smoking, diabetes, hypertension, total cholesterol, high-density lipoprotein cholesterol, triglycerides, and other five phytoestrogens. In the fully adjusted model, the urinary enterolactone (ENL) concentration was negatively correlated with NAFLD (OR of Tertile 3 : 0.48, 95% CI 0.25-0.94). When stratified by age and gender, the urinary ENL concentration was negatively correlated with NAFLD in males aged 40-59 years (OR of Tertile 3 : 0.08, 95% CI 0.01-0.82), while the urinary equol concentration was positively correlated with NAFLD in such population (OR of Tertile 3 : 4.27, 95% CI 1.02-17.85). In addition, a negative correlation between enterodiol (END) concentration and NAFLD was observed in males aged 60 years or over (OR of Tertile 2 : 0.18, 95% CI 0.05-0.69). Collectively, in middle-aged males, urinary ENL may be associated with a lower risk of NAFLD, while urinary equol may be related to a higher risk. In addition, urinary END has a possible relationship with a reduced risk of NAFLD in elder males. Definitely, clinical randomized controlled trials are needed to further verify the conclusions.

Pre- and postharvest measures used to control decay and mycotoxigenic fungi in potato (Solanum tuberosum L.) during storage.

Potato (Solanum tuberosum L.), a worldwide, staple food crop, is susceptible to postharvest rots caused by a variety of fungal pathogens, including Fusarium spp., Alternaria spp., Phytophthora infestans, Helminthosporium solani, Rhizoctonia solani, and Colletotrichum coccodes. Rots resulting from infections by these pathogens cause a significant reduction in potato quality and marketable yield. Importantly, some of these decay fungi also produce mycotoxins that represent a potential risk to human health. In the present review, an overview and discussion are provided on the epidemiology and pathogenesis of decay fungi, especially Fusarium spp., that include recent data derived from genomic and phylogenetic analyses. The biosynthesis and functional role of fungitoxic metabolites such as trichothecene mycotoxins and fusaric acid, produced in rotted potatoes are also reviewed. Advances in pre- and postharvest measures for rot management, especially eco-friendly methods including physical control, biological control, the use of natural compounds, and other agricultural management practices are also reviewed. Lastly, novel approaches to control potato dry rot such as the use of mycoviruses and CRISPR technology are highlighted.

Protective effects of lipoic acid-niacin dimers against blue light-induced oxidative damage to retinal pigment epithelium cells.

AIM: To evaluate the protective effects of lipoic acid-niacin (N2L) dimers against blue light (BL)-induced oxidative damage to human retinal pigment epithelium (hRPE) cells in vitro. METHODS: hRPE cells were divided into a control group (CG), a BL group, an N2L plus BL irradiation group, an α-lipoic acid (ALA) plus BL group, an ALA-only group, and an N2L-only group. hRPE cellular viability was detected by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) bromide assays, and apoptosis was evaluated by annexin-V-PE/7-AAD staining followed by flow cytometry. Ultrastructural changes in subcellular organelles were observed by transmission electron microscopy. Reactive oxygen species formation was assayed by flow cytometry. The expression levels of the apoptosis-related proteins BCL-2 associated X protein (BAX), B-cell leukmia/lymphoma 2 (BCL-2), and caspase-3 were quantified by Western blot analysis. RESULTS: BL exposure with a light density of 4±0.5 mW/cm2 exceeding 6h caused hRPE toxicity, whereas treatment with a high dose of N2L (100 mol/L) or ALA (150 mol/L) maintained cell viability at control levels. BL exposure caused vacuole-like degeneration, mitochondrial swelling, and reduced microvillus formation; however, a high dose of N2L or ALA maintained the ultrastructure of hRPE cells and their organelles. High doses of N2L and ALA also protected hRPE cells from BL-induced apoptosis, which was confirmed by Western blot analysis: BCL-2 expression significantly increased, while BAX and caspase-3 expression slightly decreased compared to the CG. CONCLUSION: High-dose N2L treatment (>100 mol/L) can reduce oxidative damage in degenerating hRPE cells exposed to BL with an efficacy similar to ALA.

Protection of tight junction between RPE cells with tissue factor targeting peptide.

AIM: To investigate the effect of tissue factor targeting peptide (TF-TP) on retinal pigment epithelium (RPE) cells tight junctions. METHODS: Cell counting kit-8 (CCK-8) was used to measure the proliferation of ARPE-19 cells. Expression of tight junction, ZO-1 in ARPE-19 cells was measured by Western blot and immunofluorescent staining. Western blot was also used to detect the expression of tissue factor (TF). CEC Transmigration Assay was used to measure the migration of ARPE-19 cells. The transport of fluorescent markers [fluorescein isothiocyanate dextrans of 4, 10, 20 (FD4, FD10, FD20)] and the transepithelial electrical resistance (TEER) were used to measure in ARPE-19 cell. RESULTS: CCK-8 assay showed that 5 µmol/L TF-TP can inhibit ARPE-19 cells abnormally proliferation stimulated by lipopolysaccharide (LPS; P<0.05). LPS increased the transport of fluorescent markers (FD4, FD10, FD20) and decreased TEER levels in ARPE-19 cells, respectively, which were prevented by 5 µmol/L TF-TP pretreatment (P<0.05). Furthermore, LPS significantly up-regulated the expression of TF and downregulated the expression of ZO-1 (P<0.05) in ARPE-19 cell which was inhibited by the TF-TP (P<0.05). In addition, TF-TP inhibited the abnormal migration induced by LPS in ARPE-19 cell (P<0.05). CONCLUSION: Our findings suggest that TF-TP suppressed proliferation and migration of ARPE-19 cells induced by LPS, and maintained the RPE tight junctions through inhibition of TF expression and increased expression of ZO-1.

Endoscopy-assisted vitrectomy in the anterior vitreous.

Endoscopy-assisted ocular surgery is a relatively old technique that is increasingly being recognized for its application in cases of vitreoretinal disease. This technique is especially useful when both the vitreous and retina are difficult to access because of media opacity, a small pupil, or a microcornea. In this context, the anterior vitreous is often difficult to dissect because of its complex pathological changes. This article reviews the common anatomical features and pathologies that are observed in the anterior vitreous, as well as the applications and indications of endoscopy-assisted vitrectomy in the anterior vitreous.

[Association between hepatocyte growth factor in tears and corneal haze in rabbits early after epipolis laser in situ keratomileusis].

OBJECTIVE: To study the dynamic changes of levels of hepatocyte growth factor (HGF) in tears and their association with corneal haze in rabbits early after epipolis laser in situ keratomileusis (Epi-LASIK). METHODS: Twenty-four New Zealand rabbits received Epi-LASIK with an ablation depth of 100 µm in one eye and of 150 µm in the other eye. Before and at 3, 7, 14, and 30 days after the surgery, the level of HGF in tears collected from the rabbits was measured using enzyme-linked immunosorbent assay (ELISA), and corneal haze was graded after surgery. RESULTS: In all the rabbits, corneal epithelium healing occurred in 3 to 5 days after Epi-LASIK. Corneal haze appeared 3 days postoperatively in the rabbits accompanied by increased levels of HGF in tears. At 3, 7, 14, and 30 days after the surgery, the rabbits with an ablation depth of 150 µm showed more obvious corneal haze (P<0.05) and significantly higher levels of HGF in tears than those with an ablation depth of 100 µm (P<0.05). CONCLUSION: In rabbits receiving Epi-LASIK, HGF levels in tears and the grade of corneal haze show a positive correlation early after the surgery and are both related with the depth of ablation.

Serum concentrations of cytokines in infants with retinopathy of prematurity.

Retinopathy of prematurity (ROP) is the leading cause of blindness in preterm infants. In this study, we investigated the cytokine levels in cord blood of normal preterm neonates and preterm infants developed ROP. Serum levels of 10 cytokines in umbilical cord blood were measured by multiplex protein arrays from 62 healthy preterm neonates and 30 preterm neonate cases who developed ROP at later stage. Results showed that serum levels of cytokines including interleukin 7 (IL-7), monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein 1 alpha (MIP-1α), and macrophage inflammatory protein 1 beta (MIP-1ß) were significantly increased in cases who developed ROP than in healthy preterm neonates (3.5-fold, 3.2-fold, 3.4-fold, and 2.1-fold, respectively), whereas levels of these four cytokines did not reveal any significant differences between healthy preterm infants and normal infants. When comparing the expression of cytokines in ROP patients with different clinical parameters, ROP cases whose gestational age at delivery earlier than 29.0 weeks demonstrated increased levels of MCP-1 and MIP-1ß than those later than 29.0 weeks (p < 0.05). Also, ROP cases with birth weight less than 1.28 kg revealed significantly higher level of MIP-1ß than those who were heavier than 1.28 kg (p < 0.05). These data indicated that levels of IL-7, MCP-1, MIP-1α, and MIP-1ß were associated with increased risk of ROP, in which MIP-1ß may be further correlated with the severity of ROP.

Effect of soluble inducible costimulator level and its polymorphisms on age-related macular degeneration.

Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly population. Evidence has shown that the human immune system may play critical roles in this disease. Inducible costimulator (ICOS) promotes T-cell activation, differentiation, and T:B-cell interactions. The aim of the study was to understand the effect of ICOS on the development of AMD from genetic polymorphism perspective and serum level perspective. Two ICOS polymorphisms, rs10183087A/C and rs10932037C/T, were tested in 223 AMD cases and 262 healthy controls. The serum level of soluble ICOS (sICOS) was compared among subjects with different genotypes, as well as between AMD patients and controls. Data showed that prevalence of rs10183087CC genotype was significantly increased in AMD than in controls (p=0.001). Function analysis revealed that subjects carrying rs10183087CC genotype had higher serum levels of sICOS than those with AA or AC genotypes (p<0.05). When we compared serum levels of sICOS between cases and controls, results showed that AMD patients had significantly increased sICOS levels than healthy donors (p<0.05). Also, wet type cases were observed to have higher sICOS levels than cases with dry type (p<0.05). These data suggested ICOS polymorphism could affect the susceptibility to AMD by elevating protein expression, and serum levels of sICOS may be closed correlated with the development and progression of this disease.

Endoscope-assisted goniosynechialysis combined with phacoemulsification and intraocular lens implantation to manage primary angle-closure glaucoma.

AIM: To describe and evaluate a new ophthalmic endoscope surgical technique combined with phacoemulsification and intraocular lens (IOL) implantation to treat goniosynechialysis and manage primary angle-closure glaucoma (PACG). METHODS: Endoscope-assisted goniosynechialysis combined with phacoemulsification and IOL implantation was performed in 32 eyes of 29 patients with PACG. Regular follow-up was performed 1 week and 1 month, 3, and 6 months after surgery to assess complications, intraocular pressure (IOP), anterior chamber depth, visual acuity, and anterior chamber angle. RESULTS: Preoperative mean IOP was 24.88±7.22mmHg with pharmacological treatment, and was 13.70±4.02, 13.06±3.74, 14.29±4.70, and 14.33±5.01mmHg 1 week, 1 month, 3 months, and 6 months after surgery, respectively. The postoperative decrease in IOP was significant (P<0.05). The rate for all eyes with IOP of 21mmHg or less was 93.8% (30 eyes) at the final visit without ocular hypotensive agents. The average preoperative anterior chamber depth was 1.60±0.64mm, and this value significantly increased to 2.72±0.62, 2.76±0.70, 2.73±0.68, and 2.74±0.71mm at 1 week, 1 month, 3 months, and 6 months, respectively. Visual acuity was improved in 28 eyes (87.5%) at 6 months postoperatively. The anterior chamber angle had increased in 25 eyes (78.1%) at the final visit; it was adhesive 90°-180° in 6 eyes, 180°-270° in only 1 eye. Two eyes exhibited minimal hyphema in the early postoperative period, but it could gradually be absorbed. Fibrinous reaction was observed in five eyes and spontaneously disappeared within 7 days. No shallow anterior chamber, iridodialysis, choroidal detachment, or malignant glaucoma was found in any eyes. CONCLUSION: Endoscope-assisted goniosynechialysis combined with phacoemulsification and IOL implantation to manage PACG has several advantages, including optimized visualization, greater accuracy, and improved safety. Our results suggest that it has certain curative effects and clinical application value.

Transforming growth factor-ß1 level in tears and corneal haze formation following flap-on or flap-off epi-LASIK.

OBJECTIVE: To observe the influence of flap-on or flap-off Epipolis laser in situ keratomileusis (epi-LASIK) on the release of transforming growth factor-ß1 (TGF-ß1) in tear fluid and corneal haze formation. METHODS: Thirty patients (60 eyes) with myopia underwent epi-LASIK surgery with epithelial flap repositioning (flap-on) in the right eyes and epithelial flap removal (flap-off) in the left eyes. The level of TGF-ß1 in tears was measured preoperatively and on days 1, 3, and 7 postoperatively. Corneal haze was graded at 1, 3 and 6 months after surgery. RESULTS: The mean preoperative spherical equivalent refraction was -4.98∓2.28 D (-2.50 to -7.25 D) in flap-on group and -5.20∓4.02 D (-1.75 to -7.00 D) in flap-off group, showing no significant difference between the two groups (P=0.80). TGF-ß1 levels in the tear fluid were similar in the two groups preoperatively (P=0.11) and at 1, 3, and 7 days postoperatively (P=0.55, 0.45, 0.19, respectively). TGF-ß1 levels in tears gradually decreased after the first postoperative day in both groups, but were still higher than the preoperative value till the 7th postoperative day. Corneal haze scores in the two groups were similar at 1 month (P=0.98), 3 months (P=0.52), and 6 months (P=0.72) after the operation. CONCLUSION: Flap-on and flap-off epi-LASIK surgeries do not differ significantly in postoperative TGF-ß1 levels in the tear fluid or in the postoperative haze scores. TGF-ß1 may play a role in corneal wound healing.

Effects of lysyl oxidase genetic variants on the susceptibility to rhegmatogenous retinal detachment and proliferative vitreoretinopathy.

Proliferative vitreoretinopathy (PVR), the most common cause of failure of rhegmatogenous retinal detachment (RD) surgery, is an anomalous scarring process related to ocular inflammation. Lysyl oxidase (LOX) is a copper-dependent amine oxidase that may play important roles in ocular tissue integrity. The aim of this study was to investigate whether polymorphisms in the LOX gene were associated with susceptibility to RD and PVR. We screened the promoter region of LOX gene and tested two previously reported polymorphisms (-22 G/C and 473 G/A) in RD patients with or without PVR and healthy controls. Data showed that prevalence of the -22CC genotype and -22C allele were significantly higher in the RD cases than in the control group after adjustment for sex and age (p < 0.001 and p < 0.001, respectively). Similarly, a significant difference was observed regarding LOX 473GA genotype and 473A allele between RD patients and healthy donors after adjustment for sex and age (p = 0.005 and p = 0.012, respectively). Also, when compared to RD cases without PVR, patients who developed PVR had significantly higher numbers of -22CC genotype and -22C allele (p = 0.048 and p = 0.003, respectively). These results indicated that LOX polymorphisms were associated with increased susceptibility to RD and PVR and suggest a potential correlation between LOX and ocular inflammation.

[Endoscope-assisted goniosynechialysis with phacoemulsification and intraocular lens implantation for treatment of primary angle-closure glaucoma].

OBJECTIVE: To evaluate the early therapeutic effect of endoscope-assisted goniosynechialysis combined with phacoemulsification and intraocular lens implantation for management of primary angle-closure glaucoma. METHODS: Endoscope-assisted goniosynechialysis combined with phacoemulsification and intraocular lens implantation was performed in 31 patients (31 eyes) with primary angle-closure glaucoma, including 17 eyes with acute angle closure glaucoma and 14 eyes with chronic angle closure glaucoma. The patients were regularly followed up to observe the postoperative complications and changes in the intraocular pressure, visual acuity, anterior chamber depth and anterior chamber angle. RESULTS: Hyphema occurred in 3 eyes after the operation and was absorbed gradually. The intraocular pressure decreased significantly in all the patients after the operation (P<0.05) but showed no significant differences between the acute and chronic cases (P>0.05). The intraocular pressure was over 21 mmHg in only one eye in the chronic group, and could be controlled by ocular hypotensive agents. Visual acuity improved and the anterior chamber depth increased in all the patients, showing no significant differences between the two groups (P>0.05). The anterior chamber angle all opened after the operation in 16 eyes (94.1%) in acute group, a rate significantly higher than that in the chronic group (9 eyes, 64.3%) (P<0.05). Anterior chamber angle was adhesive in 1 eye in the acute group and in 5 eyes in the chronic group. No further extension of goniosynechiae was found in these cases during the follow-up period. CONCLUSIONS: Endoscope-assisted goniosynechialysis has such advantages as optimized visualization, accuracy and safety in the management of primary angle-closure glaucoma.

Long-Term in vitro expansion of retinal progenitor cells by culturing intact neurospheres in monolayer.

This study developed a modified monolayer culture technique for expansion of retinal progenitor cells in vitro. Using this method, spheres collected by low centrifugation from free floating cultures were re-seeded onto poly-D-lysine pre-coated flasks, and cultured in the retinal progenitor cells medium (including fibrous growth factor, epidermal growth factor and leukemia inhibitor factor) plus 5% FBS for the first 24 h, then FBS was removed. These cultured cells were proliferative, and in addition to expressing the neuroectodermal marker nestin they were multipotential. By immunocytochemistry and FACS analysis, we demonstrated that the percentage of nestin-expressing cells in passage 2 and passage 8 was 92.5 and 85.5%, respectively, which was greater than the percentage of their counterparts, 81.9 and 76.2%, in simple adhesive cultures (p < 0.05). Moreover, these cells preferentially differentiated into retinal neurons rather than glial cells.

Topical anesthesia in transconjunctival sutureless 25-gauge vitrectomy for macular-based disorders.

PURPOSE: To evaluate the safety and feasibility of topical anesthesia in vitrectomy using the transconjunctival sutureless 25-gauge system (TSV 25G). METHODS: TSV 25G vitrectomy was performed prospectively in 46 eyes of 46 patients with macular-based disorders, including macular holes (n = 31), idiopathic epiretinal membranes (n = 11), and vitreoretinal traction syndrome (n = 4). Topical anesthesia was administered in each patient by instilling 2% Alcaine at 5 min intervals, three times before surgery. Additional Alcaine drops were given as a supplementation if needed during the operation. The levels of intraoperative analgesia were graded from 1 (adequate) to 3 (inadequate). RESULTS: Thirty-one patients (67.4%) tolerated the procedure well, with no additional anesthetic agent. Thirteen patients (28.3%) required additional topical anesthesia and 2 patients (4.3%) required a systemic sedative. Intraoperative pain occurred when the sclera was pierced by the microcannula and when sclera indentation was performed. No patient required additional retrobulbar or peribulbar anesthesia. There was no intraoperative complication which was specifically related to topical anesthesia. CONCLUSION: Topical anesthesia permits the successful management of some surgically less complex vitreoretinal disorders when the TSV 25G is used.

[The preliminary study of photolysis for cataract surgery].

OBJECTIVE: To evaluate the safety and efficiency of Dodick-ARC photolysis machine (a kind of Nd:YAG laser system) in the using of cataract surgery. METHODS: 52 eyes of cataract (fifty-two patients) were undergone photolysis with Dodick-ARC laser system and foldable intraocular lenses (IOLs) implantation. The hardness of nuclei was graded as N(1.0) to N(4.0) of LOCS III classification system. Before and after operations, the intraocular conditions, visual acuity, corneal thickness and operative complications were recorded. The time of follow-ups was more than 6 months. RESULTS: The operations were accomplished in 28 eyes and were changed into Phacoemulsification in 24 eyes. Posterior capsule rupture occurred in 2 cases (3.8%). Severe corneal edema was found in 3 cases (5.8%) that disappeared in 1 to 2 weeks. Preoperative visual acuity was < 0.05 in 19 cases, 0.05 - 0.3 in 28 cases and 0.4 - 0.5 in 5 cases. Postoperative visual acuity was < or = 0.3 in 1 case, 0.4 - 0.5 in 12 cases, 0.6 - 0.9 in 25 cases and > or = 1.0 in 14 cases. The mean values of pachymetry was 0.59 mm preoperatively and 0.60 mm in 3 months postoperatively (P = 0.097). CONCLUSION: Photolysis using Dodick-ARC Nd:YAG laser system is a safe and effective method for cataract surgery. The further improvement of equipment and surgery techniques is needed.