Hypertriglyceridemic waist phenotype: Association with initial neurological severity and etiologic subtypes in patients with acute ischemic stroke.

Objective: To explore the relationship of hypertriglyceridemic waist phenotype (HTWP) with initial neurological severity and etiologic subtypes in patients with acute ischemic stroke. Methods: The data for this study were collected from hospitalized patients within 72 h of acute ischemic stroke onset at the Department of Neurology of the Affiliated Hospital of Beihua University from 1 July 2020 to 30 June 2022. The initial neurological severity was assessed by the National Institute of Health Stroke Scale (NIHSS) on the day of admission: NIHSS <6 was defined as mild stroke, and NIHSS ≥6 as moderate to severe stroke. HTWP was defined by fasting serum triglycerides ≥1.7 mmol/L and waist circumference ≥90 cm in men and ≥80 cm in women. Differentiation of etiologic subtypes was based on the method reported in the Trial of Org 10 172 in Acute Stroke Treatment. Multivariate logistic regression analysis was used to analyze the association of HTWP with initial neurological severity and etiologic subtypes. Results: The study included 431 patients. Compared with the normal waist-normal blood triglyceride group, patients with HTWP had reduced risks of moderate to severe stroke [odds ratio (OR): 0.384, 95% confidence interval (CI): 0.170-0.869; P = 0.022]. In addition, the risk of small-artery occlusion stroke was 2.318 times higher in the HTWP group than in the normal triglyceride-normal waist (NWNT) group (OR: 2.318, 95% CI: 1.244-4.319; P = 0.008). Conclusion: Initial neurological severity was less severe in patients with HTWP, and HTWP was associated with an increased risk of small-artery occlusion stroke.

Discriminating Eggs from Two Local Breeds Based on Fatty Acid Profile and Flavor Characteristics Combined with Classification Algorithms.

This study discriminated fatty acid profile and flavor characteristics of Beijing You Chicken (BYC) as a precious local breed and Dwarf Beijing You Chicken (DBYC) eggs. Fatty acid profile and flavor characteristics were analyzed to identify differences between BYC and DBYC eggs. Four classification algorithms were used to build classification models. Arachidic acid, oleic acid (OA), eicosatrienoic acid, docosapentaenoic acid (DPA), hexadecenoic acid, monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), unsaturated fatty acids (UFA) and 35 volatile compounds had significant differences in fatty acids and volatile compounds by gas chromatography-mass spectrometry (GC-MS) (p<0.05). For fatty acid data, k-nearest neighbor (KNN) and support vector machine (SVM) got 91.7% classification accuracy. SPME-GC-MS data failed in classification models. For electronic nose data, classification accuracy of KNN, linear discriminant analysis (LDA), SVM and decision tree was all 100%. The overall results indicated that BYC and DBYC eggs could be discriminated based on electronic nose with suitable classification algorithms. This research compared the differentiation of the fatty acid profile and volatile compounds of various egg yolks. The results could be applied to evaluate egg nutrition and distinguish avian eggs.

Surgical management of fungal endophthalmitis resulting from fungal keratitis.

AIM: To report the fungal organisms, clinical features, surgical treatment strategies, and outcomes of patients with culture-proven exogenous fungal endophthalmitis (EFE) secondary to keratitis, and evaluate the role of surgery in the treatment. METHODS: The clinical records of 27 patients (27 eyes) with culture-proven EFE resulting from fungal keratitis treated at Shandong Eye Institute from January 2007 to January 2015 were retrospectively reviewed. Information about fungal culture results, clinical features, surgical procedures, and final visual acuity was obtained. RESULTS: There were 39 positive culture results from samples of cornea, hypopyon, vitreous and lens capsule, accounting for 56%, 26%, 15% and 2.5%, respectively. Fusarium was identified in 44% (12/27) of the eyes, followed by Aspergillus in 22% (6/27). Posterior segment infection was involved in 78% (21/27) of the patients. The corneal infection was larger than 3 mm ×3 mm in 89% (24/27) of the patients, and 22% (6/27) of them had the entire cornea, and even the sclera involved. Three eyes had silicone oil tamponade, and two eyes had retinal detachment. Twenty-two eyes (81.5%) underwent penetrating keratoplasty (PKP), and over half of them (54.5%) were operated within 3d from the onset of antifungal therapy. Fourteen eyes (52%) underwent intracameral antifungal drug injection, and three of them required repeated injections. Fifteen eyes (55.6%) underwent pars plana vitrectomy (PPV). The rate of the eyes undergoing PPV as the initial surgical procedure was 60% (9/15), lower than 77% in PKP. Intravitreal injection was given in 59% of the eyes (16/27), and 75% of them required repeated injections. The final visual acuity was 20/100 or better in 37% of the eyes, and better than counting fingers in 55.6% of the eyes. Five eyes (18.5%) were eviscerated. In the two eyes with concurrent retinal detachment, one achieved retinal reattachment, and the other was eviscerated. In the three eyes with silicone oil tamponade, two eyes received silicone oil removal, and the other one was eviscerated. CONCLUSION: Fusarium and Aspergillus are the dominant pathogens in EFE resulting from keratitis. Aggressive antifungal surgeries including multiple intravitreal injections, PKP and core vitrectomy (especially in the initial surgery) are helpful procedures to improve prognosis of severe EFE secondary to keratitis.

[The expression and function of annexin A2 in the course of retinal angiogenesis of mouse].

OBJECTIVE: To study the production and the role of annexin A2 (ANXA2) in the process of retinal neovascularization of mouse. METHODS: Experimental study. C57BL/6J mice were classified into four groups:normal group (80 mice), oxygen induced retinopathy (OIR) mock group (80 mice) , siANXA2 group (transfected with siRNA target ANXA2) (50 mice) , and siANXA2_M group (50 mice) .Stretched preparation of retina after angiography was used to observe the morphology change of retinal neovascularization from 12 to 30 days after birth in normal group and OIR mock group, and real-time PCR was used to test the expression of ANXA2 in these days. On 17 days old, the mRNA and protein production of vascular endothelial growth factor (VEGF)-α, matrix metalloproteinase (MMP)-2, MMP-9, and tissue inhibitor of metalloproteinase (TIMP)-2 in 4 groups were assessed by real-time PCR and Western blot. The results of all factors among four groups were analyzed by one way ANOVA and SNK-q test. RESULTS: The retinal neovascularization of siANXA2 group in 17 days old was more regular than that in OIR mock group. The production of ANXA2 in mouse retina was associated with the stage of retinal neovascularization. The expression of ANXA2 was in high level when neovessels grew and in low level when neovessels stopped growing. The mRNA expressions of ANXA2,VEGF-α, MMP-2, MMP-9 and TIMP-2 showed statistical difference among 4 groups (F = 8.122-74.009, P < 0.05) . Significant statistics difference was found in multiple comparison:the expressions of VEGF-α (0.22 ± 0.04), MMP-2 (11.08 ± 1.28), MMP-9 (4.64 ± 0.38) in OIR mock group were significantly higher than that in normal group (0.16 ± 0.02, 2.18 ± 1.39, 1.17 ± 0.25) (SNK-q test: P < 0.01).In siANXA2 group, the productions of VEGF-α (0.02 ± 0.01), MMP-2 (2.21 ± 0.42) , MMP-9 (1.33 ± 0.10) were significantly lower than that in OIR mock group (SNK-q test: P < 0.01). The mRNA expression of TIMP-2 (0.59 ± 0.15) in OIR mock group was significantly lower than that in normal group (1.35 ± 0.01) (SNK-q test: P < 0.05). In siANXA2 group, the production of TIMP-2 was higher than that in OIR mock group (SNK-q test: P < 0.05). The results of Western blot were similar to that in real-time PCR. CONCLUSIONS: ANXA2 is overexpressed in oxygen-induced retinal neovascularization in a mouse model. The overexpression of ANXA2 may affect the expression of proangiogenic factors. ANXA2 may involve in the development of the retinal neovascularization. The production of ANXA2 may be inhibited by siRNA. ANXA2 maybe a new target for inhibition of retinal neovascularization.

Black diaphragm intraocular lens implantation and penetrating keratoplasty in aphakic eyes with traumatic aniridia.

AIM: To evaluate the long-term outcome of implantation of black diaphragm intraocular (BDI) lens combined with penetrating keratoplasty (PKP) for managing aphakic eyes with traumatic aniridia and corneal damage. METHODS: Six aphakic eyes of six patients with traumatic aniridia and corneal damage had BDI lens implantation at Qingdao Eye Hospital, Shandong Eye Institute from June 2008 to November 2011. Medical records of the patients were reviewed. Three patients received PKP and after 12-18months were implanted with BDI lens. The other three patients completed PKP and BDI lens implantation at the same time. The corrected visual acuity, intraocular pressure and number of corneal endothelial cells were monitored. RESULTS: The patients were followed up for an average of 24.3±12.1months (range 14-48 months). All BDI lenses were located well. The best corrected visual acuity got improved in 5 patients (0.1-1.0) and decreased in 1 patient from 0.4 to 0.2. Three patients had normal intraocular pressure (IOP) after implantation. Two patients required antiglaucoma medications to control IOP within the normal range and 1 patient implanted Ahmed glaucoma valve to control IOP. The corneal grafts kept transparent in all eyes and the corneal endothelial counting >1 000/mm(2), although two patients experienced acute graft rejection and loss more than 30% corneal endothelial cells. CONCLUSION: Implantation of BDI lens combined with PKP is an effective option for managing aphakic eyes with traumatic aniridia and corneal damage. Although the results in our study are encouraging, additional studies of the long-term safety and efficacy are required. A larger study population and longer follow-up may be beneficial.

Corneal endothelial cell density and morphology in Chinese patients with pseudoexfoliation syndrome.

AIM: To evaluate the corneal endothelial cell density and morphology in Chinese patients with pseudoexfoliation syndrome (PEX). METHODS: Medical records of 16 patients (20 eyes) with PEX who presented to our institution between July 2008 and June 2010 were retrospectively reviewed. Thirteen eyes had combined glaucoma. The information of five apparently normal fellow eyes in these patients was also recorded. Left eyes of 20 patients with bilateral senile cataracts but no other eye disease were included as controls. Specular microscopy was performed in all eyes to analyze for corneal endothelial cell density and morphology. Cell density, coefficient of variation in cell size, and percentage of hexagonal cells in corneal endothelium were evaluated. RESULTS: The mean corneal endothelial cell density in the PEX eyes was 2298±239 cells/mm(2), significantly lower than that in the cataract eyes (2652±18 cells/mm(2), P=0.026), but there were no significant differences in coefficient of variation of cell size and frequency of hexagonality between these two groups. No significant differences in the three parameters were found between the apparently normal fellow eyes and the PEX eyes or the cataract eyes, or between the PEX eyes with and without glaucoma. CONCLUSION: Corneal endothelial cell density may decrease in Chinese patients with PEX. The development of glaucoma in PEX eyes does not seem to be related with the change in corneal endothelial cell density or morphology.

[Experimental study of (Pro)renin receptor siRNA inhibiting retinal neovascularization].

OBJECTIVE: To observe the inhibit effect of (pro)renin receptor siRNA on mice retinal neovascularization, and investigate its possible mechanism. METHODS: Experimental study. 72 new born C57BL/6J mouse were randomly divided into six groups: group A to group E were exposed to hyperoxia, and then returned to normoxia to induce retinal neovascularization. Group A were treated with PRR siRNA plasmid, group B with control plasmid, group C with PRR siRNA and Losartan, and group D with Losartan. Group E were not treated. Group F was control group. Mouse were sacrificed at postnatal day 17, retinal perfusion stretched preparation and HE dyeing method were used to observe the status of retinal neovascularization. PRR expression and the activation of extracellular regulated protein kinases 1/2 (ERK1/2) were detected by Western Blot. And Real Time PCR was used to detect the expression of transforming growth factor-ß1 (TGF-ß1) in group A, B, E and F. Analysis of one way variance (LSD) was used and statistical difference was considered significant at a P value less than 0.05. RESULTS: The mouse treated with PRR siRNA, Losartan and combined therapy could significantly reduce retina neovascularization and vessel leakage compared with oxygen-induced retinopathy group and control plasmid group. Average counts of vascular endothelial cells which break through the inner limiting membrane performed at postnatal day 17 in PRR siRNA group (4.47 ± 1.96), Losartan group (5.94 ± 2.54) and combined therapy group (4.49 ± 2.53) were significantly lower than oxygen-induced retinopathy group (32.73 ± 6.38) (P < 0.05) and control plasmid group (21.04 ± 5.39). Western Blot showed that PRR protein express in hyperoxia induced group was significantly higher than normal (P = 0.007). After treated with PRR siRNA or combined therapy, the expression of PRR protein was significantly lower than hyperoxia induced group (P < 0.05). There are no significantly differences between control plasmid group, Losartan group and hyperoxia induced group (P > 0.05). The activated ERK1/2 lever in hyperoxia group was significantly higher than normal (P = 0.003). After treated with PRRsiRNA, Losartan or combined therapy, activated ERK1/2 lever was significantly lower than hyperoxia induced group (P < 0.05). And the effect of PRR siRNA group and combined therapy group seems more obviously, compared with Losartan group, the difference was significantly (P < 0.05). Real Time PCR showed that the lever of TGF-ß1 in hyperoxia group was significantly higher than normal (P = 0.001). After treated with PRR siRNA, the TGF-ß1 was significantly reduced (P = 0.004), and there was no significantly difference between control plasma group and hyperoxia induced group (P = 0.222). CONCLUSIONS: PRR combined with prorenin or renin could activate ERK1/2 signal transduction passageway, and promote cell proliferation, differentiation and migration, thus promote retinal neovascularization. PRR siRNA could obviously reduce PRR expression, inhibit ERK1/2 signal transduction passageway activation, and diminish retinal neovascularization.

[Experimental study of Twist siRNA to inhibit the formation of retinal neovascularization in mice].

OBJECTIVE: The purpose of this research is to find the law of neovascular endothelial cell migration and transition through repressing the expression of Twist in mouse's retinal neovascularization with RNAi, and get a new target of inhibit retinal neovascularization. METHODS: Oxygen-induced retinopathy (OIR) was produced in new born C57BL/6J mice by exposing postnatal day 7 (P7) pups to 75% oxygen for 5 days. P12 pups were injected 1 µl pTwist. siRNA plasmid solution or 1 µl negative siRNA plasmid solution into vitreous cavity. Eyeballs were enucleated for Evans blue angiography, histopathologic examination, neovascular endothelial cell counting, immunohistochemistry and Real-Time PCR. RESULTS: Observed by light microscopy retinal neovascularization, the number of vascular endothelial cells per eye were 0.34 ± 0.11, 32.73 ± 6.38, 4.56 ± 2.02 and 20.17 ± 6.49 in the normal control group, hyperoxia group, Twist plasmid group and the control plasmid group. Mouse retinal Evans blue perfusion and HE staining of paraffin sections showed that retinal vascular leakage, tortuous and angiogenesis significantly reduced in Twist plasmid group compared with hyperoxia group. Endothelial cell count was significantly decrease in Twist plasmid group. Both immunohistochemistry and real time PCR proved that Twist and vimentin expression in hyperoxia group were significantly higher than that of Twist plasmid group (F = 27.214, 31.211;P < 0.05). CONCLUSION: As mice retinal neovascular growth, Twist may play important roles as a cell transition regulatory factor. Repressing Twist with RNAi, we can repress cell transition and inhibit retinal neovascular.

[Progress on studies of exfoliation glaucoma].

Exfoliation glaucoma is a specific type of glaucoma secondary to exfoliation syndrome. Exfoliation syndrome is one of the most common identifiable causes of secondary open-angle glaucoma worldwide. In recent years, great progress has been made in exfoliation syndrome and exfoliation glaucoma. The study of exfoliation glaucoma in the aspects of prevalence, inheritance, formation of exfoliative material, gene analysis and treatment are reviewed in this article.

[Severe fungal corneal ulcer resulting in infectious endophthalmitis].

OBJECTIVE: To analyze causes of serious fungal corneal ulcer resulting in infectious endophthalmitis and explore clinical strategies of avoiding the failure of antifungal therapy. METHODS: Etiological factors, pre-hospital treatments, clinical features and laboratory findings of 47 inpatients with fungal corneal ulcer resulting in endophthalmitis from January 1999 to December 2008 in Qingdao eye hospital were retrospectively reviewed. RESULTS: Rural residents (95.7%) dominated in 47 cases with a mean age of (49.8 ± 10.1) years. Ocular trauma was the leading cause of fungal corneal ulcer (66.0%). Three patients were ever treated with hormone drugs after the fungal infection. Primary, secondary and tertiary hospital accounted for 68.1%, 17.0% and 14.9% among first medical consultation sites. Diagnostic accuracies of fungal corneal ulcer in three grade hospitals were 31.3%, 62.5% and 71.4% respectively. The average interval from the onset of disease to the admission into our hospital was (29 ± 23) days. The dominating pathogen was genus Fusarium (91.5%) with F. solani (48.9%), F. oxysporum (31.9%) and F. moniliforme (8.5%). Antifungal drug sensitivity tests were performed in 21 patients. The first three sensitive drugs were natamycin (88.9%), voriconazole (78.6%) and amphotericin B (61.9%). The first three drug-resistant ones were miconazole (90.5%), fluconazole (66.7%) and itraconazole (61.9%). CONCLUSION: Main causes of fungal corneal ulcer resulting in infectious endophthalmitis included lower diagnostic accuracies of first medical consultation in primary hospitals, abuses of non-sensitive drug and delayed treatment of patients. Improving clinical capabilities of doctors in primary hospitals, emphasizing antifungal drug susceptibility tests, and consummating the social security system and the referral system could be effective measures to avoid therapeutic failures.

[Expression of mouse telomerase reverse transcription in a mouse model of oxygen-induced retinopathy].

OBJECTIVE: To establish oxygen-induced retinal neovascularization in mice and to detect the expression of mTERT in mice. METHODS: It was an experimental study Establishment of oxygen-induced retinal neovascularization in mice. Thirty-two 7-day-old C57BL/6J mice were divided into oxygen-induced retinopathy group and control group without restriction of gender. In oxygen-induced retinopathy group, 16 mice were exposed to 75% +/- 2% oxygen for 5 days and then to room air; In control group, 16 mice were raised in room air. Observation of the retinal neovascularization. On the postnatal day 19, The mice's vena caudalis were perfused with 2% Evens blue solution. Eyeballs were enucleated and fixed in 4% paraformaldehyde for half an hour. Then the retina was separated and flat-mounted on the slide. The morphologic changes of retinal vessel were observed and captured under fluorescence microscope. Histological observation and vascular endothelial cells counting. The eyeballs were enucleated and then fixed. After paraffin imbedding, 4 microm serial slices, hematoxylin-eosin staining, select one section every 60 microm to count the endothelial cell nucleus that break through the inner limiting membrane. Expression of mTERT mRNA were confirmed by reverse-transcription polymerase chain reaction (RT-PCR). In the each group, the retina were all carefully dissected on the postnatal day 19. The total RNA was isolated and cDNA was synthesized before RT-PCR was performed. The PCR products were separated by 2% agarose gel electrophoresis and photographed. Expression of mTERT mRNA were confirmed by Real-time PCR The total RNA was isolated and cDNA was synthesized (The same procedure as RT-PCR). Fluorescent real-time quantitative polymerase chain reaction system (total 20 microl) was made. The Fluorescent signals were detected at 60 degrees C. The expression of mTERT were confirmed by immunohistochemistry. At P19, 4 microm cross sections were made in the hyperoxia-exposed and normal retinas. Sections were incubated with rabbit anti-Human/Mouse/Rat Telomerase 60 minutes at 37 degrees C. Anti-rabbit immunoglobulin G, depending on the primary antibody, was used as a secondary antibody for 30 min. Peroxidase activity was detected with the substrate diaminobenzidine. Permanent slides were covered with a 1.5 mm thick cover slip, examined using a light microscope and photographed. RESULTS: The central retina was nonperfused region at P12. The most of the central retina showed almost no perfusion and the radial vessels appeared tortuous and dilated at P14. Retinal neovascularization occurred at maximum between postnatal day 17 and postnatal day 19. Paraffin tissue slice with hematoxylin-eosin staining showed that in the control group the average counts of vascular endothelial cells which break through the inner limiting membrane were hardly seen, but in hyperoxia group were noticeably more than in the control group. Reverse-transcription polymerase chain reaction (RT-PCR) results: the mRNA of mTERT and bFGF in the retinopathy group were higher than in the control group (P < 0.05). Real-time PCR results: the expression of mTERT mRNA in the retinopathy group was noticeably higher than in the control group (F = 173.104, P < 0.05). Immunohistochemical staining showed that mTERT protein were positive in the retinal neovascularization of the hyperoxia group, but were negative in the retinal vessel of the control group. CONCLUSIONS: Telomerase reverse transcription and angiogenic correlation factors were up-regulated in a mouse model of oxygen-induced retinopathy, which may have therapeutic potential in the treatment with the neovascularization in retinopathy.

[Long term complications of black diaphragm aniridia intraocular lens implant in traumatic aniridia].

OBJECTIVE: To analyze long-term complications of black diaphragm aniridia intraocular lens (IOL) implant in traumatic aniridia and to investigate the causes and precautionary measures. METHODS: This is a retrospective consecutive case study. Five traumatic aniridia cases undertaken black diaphragm aniridia IOL implantation in Shandong Eye Institute and Hospital and developed severe complications during long-term follow-up were analyzed, including 4 males and 1 female, averaged 26.8 years old. The follow-up time varied from 42 months to 108 months. Two cases had implantation of a secondary black diaphragm IOL after pars plana vitrectomy. Two cases had implantation of a black diaphragm IOL together with cataract extraction. One case implanted a black diaphragm IOL only. RESULTS: All patients felt well within a short period after the surgery, symptoms of glare and photophobia were improved. A better visual acuity was obtained in a short-term period. However, severely secondary glaucoma and bullous keratopathy occurred in the long-term follow up. Visual acuity decreased to counting finger or hand motion. All cases received penetrating keratoplasty and IOL explantation. CONCLUSIONS: Black diaphragm aniridia intraocular lens implantation may induce severe long-term complications. The indications should be selected seriously and closely follow-up is important.

[TERT-siRNA inhibits oxygen-induced retinal neovascularization in mice].

OBJECTIVE: To investigate the inhibitory effect of small interfering RNA (siRNA) targeting TERT on murine retinal neovascularization and explore the feasibility of potential therapeutic approach in retinal vascular disease. METHODS: Two recombinant plasmids TERT siRNA (pSIREN-mTERT-1) and negative plasmid (pSIREN-mTERT-N) were constructed and 80 seven-day-old C57BL/6J mice were divided randomly into therapeutic group (A), negative plasmid group (B), oxygen-induced retinopathy group (C) and normal control group (D), 20 mice in each group. Group A, B and C were exposed to 75% +/- 2% oxygen for 5 days and then to room air, which induced mice retinal neovascularization. Groups A and B were injected two kinds of the above recombinant plasmid into the murine vitreous on the 12th day. The mice of group D were raised in normal oxygen circumstance. On the 19th day, 2% Evens blue angiography was used to observe the pattern of the retinal vascular. Expression of TERT mRNA were confirmed by reverse-transcription polymerase chain reaction (RT-PCR) and Real-time PCR. Histological observation and vascular endothelial cells counting were used to examine the effects of siRNA on the retinal neovascularization. RESULTS: Retinal flat after Evans blue angiography indicated that the vessels of group A formed a fine radial branching pattern, which was similar to normal mice. In group A, the retinal neovascularization reduced and the structure of retina were more regular than group B and C. At the same time the large vessels were distorted, neovascular clusters proliferated and fluorescence leaked in the middle and periphery area in group B and C. RT-PCR and Real-time PCR showed the expression of TERT mRNA was downregulated in group A compared with groups B and C (P < 0.05). Paraffin tissue slice with hematoxylin-eosin staining showed that the average counts of vascular endothelial cells which break through the inner limiting membrane in group A were less than groups B and C, the differences were significant (P < 0.05). CONCLUSION: Pathologic retinal neovascularization can be inhibited by specific TERT siRNA in vivo, which may be a novel efficient strategy against proliferative vasculopathies.

[The mechanism of twist gene regulation during the retina angiogenesis].

OBJECTIVE: To study the mechanisms of the Twist gene and cell transition during the angiogenesis in a mouse model of oxygen induced retinopathy. METHODS: It was a experimental study. 40 new born C57/BL mice were divided into two groups; the control group, which were fed in the room air, and the study group, which were fed 7 days in the normal environment, followed by exposure to hyperoxia (75% O2) for 5 days. The mice were sacrificed at post born 12 day and post born 17 day. The Twist and VE-cadherin expression were detected with immunohistochemistry. The total retinal RNA was extracted and the expression level of the VE-cadherin, vimentin and twist were examined with RT-PCR method. RESULTS: Immunohistochemistry proved there was no significant difference at post born 12 day. At post born 17 day, compared with the control group (75.36 +/- 7.04),VE-cadherin expression of hyperoxia group (65.19 +/- 8.39) decreased (F =8.616, P =0.009). Compared with the control group (82.14 +/- 6.32), Vimentin expression of hyperoxia group (95.09 +/- 14.13) increased, compared with the control group (93.30 +/- 6.37), Twist expression of hyperoxia group (119.48 +/- 7.90) increased (F = 66.557, P = 0.000) significantly. RT-PCR examination revealed there exist the interaction between Vimentin, Twist expression and the detection time (F = 5.508, P = 0.032; F = 17.760, P = 0.001; respectively). At post born 12 day, compared with the control group (0.77 +/- 0.10), VE-cadherin expression of hyperoxia group (0.64 +/- 0.09) decreased (P = 0.047). Compared with the control group (0.24 +/- 0.05), Vimentin expression of hyperoxia group (0.39 +/- 0.09) increased (P = 0.033). At post born 17 day, compared with the control group (0.75 +/- 0.12), VE-cadherin expression of hyperoxia group (0.51 +/- 0.07) decreased more obviously (P = 0.002), compared with the control group (0.36 +/- 0.06), Vimentin expression of hyperoxia group (0.70 +/- 0.14) increased (P = 0.000). Compared with the control group(0.89 +/- 0.11), Twist expression of hyperoxia group (1.24 +/- 0.15) increased (P = 0.003). CONCLUSION: Twist, as a cell transition gene, participated in the angiogenesis of the oxygen induced retinopathy, and that the Twist induced endothelium-mesenchymal transition may be one of the main reasons.

[Virulence of extracellular phospholipase B of Candida albicans in rabbit experimental keratomycosis].

OBJECTIVE: To determine the virulence of extracellular phospholipase B (PLB) of Candida albicans in experimental keratomycosis. METHODS: It was an experimental study. The PLB-deficient mutant strain of Candida albicans and its isogenic parental strain were used in this study. The effects of these two strains on the model of experimental keratomycosis in 48 New Zealand albino rabbits covered with contact lens was compared by observing the dynamic changes clinically and histopathologically. In vitro, these two strains were incubated with the corneal stromal cells separately (37 degrees C, 5% CO2). The influence of these two strains on monolayer keratocytes were detected by scanning electron microscopy (SEM), enzyme linked immunosorbent assay (ELISA), and flow cytometry with Annexin V/propidium iodide. RESULTS: The hyphae of these two strains grew perpendicularly to the corneal stromal lamellae. The difference of the hyphal invasion inoculated for 2 days by these two isogenic strains was statistically significant (P = 0.002), but at other intervals, no significant difference was found. The severity of the inflammation in parental keratomycosis was the same as that in PLB null strain at any time points (P > 0.05). Under SEM, the morphogenesis and the number of adherent germ tubes of these two isogenic strains appeared similarly (P > 0.05), but the number of germ tubes penetrating cell monolayer was significantly different (P = 0.009). Obviously more prostaglandin E2 (PGE2) was detected in the culture supernatants of parental strain group (65,466 +/- 5773) pg/ml than that of the null strain group (18,025 +/- 5232) pg/ml. The percentages of the cells with damaged cellular membrane in the parental group, the PLB null group and the control group, were 3.02%, 2.04% and 0.12%, respectively. The percentages of apoptosis cells in these three groups were 33.17%, 27.56% and 1.46%, respectively. The percentages of living cells were 63.81%, 70.40% and 98.41%, respectively. CONCLUSIONS: PLB shows virulent effects in triggering fungal invasion in cornea immediately following fungal adherence by decomposing membrane phospholipids and leading to cell lysis. However, its virulent effect does not appear to be critical as in the hematogenous model of disseminated candidiasis.

[The progress of epithelial-mesenchymal transition in ophthalmology].

Epithelial-mesenchymal transition (EMT), a mechanism of cell transdifferentiation, plays a key role in embryonic development, tumor metastasis and tissue reparative process. The main manifestations of EMT are disappearance of epithelium polarity, lose of cell-cell junction, increased expression of mesenchymal phenotype and activated migratory ability of epithelium. EMT existed in the cornea epithelium regeneration, fibrosis, and wound healing. EMT also participates in the pathological development of lens trauma, cataract and proliferative vitreous retinopathy. This article outlined the recent progress of EMT in ophthalmology.

[The roles of gelatinases in pathological changes of fungal keratitis in experimental rabbits].

OBJECTIVE: To study the roles of gelatinases, including matrix metalloproteinase (MMP)-2 and MMP-9, in pathological changes of fungal keratitis in rabbits. METHODS: Eighty New Zealand albino rabbits were randomly and equally divided into 4 groups, 3 of them were test groups, with Fusarium solani, Aspergillus fumigatus, and Candida albicans inoculated onto the right corneas, respectively. In the other group, sterile saline was injected onto the right corneas and was used as the control group. The source and activity of gelatinases were examined by immunohistochemistry and gelatin zymography, respectively. Hematoxylin-eosin stains was applied for observing infiltration of inflammatory cells and degradation of corneal extracellular matrixes (ECMs). Periodic acid-Schiff stain was used for studying hyphal growth patterns and the invasive depth in the cornea. RESULTS: MMP-2 was mainly produced by the keratocytes. Active MMP-2 was detected from day 5 after inoculation and increased greatly on day 8. MMP-9 was mostly produced by neutrophils, and active MMP-9 was detected from day 1 and increased to day 3, then decreased gradually. In A. fumigatus and C. albicans keratitis, the corneal ECMs were degraded obviously, the hyphae grew vertically, and the neutrophils were much more than those in F. solani keratitis whose hyphae grew horizontally and ECMs were degraded slightly. On day 8, the hyphae and neutrophils in F. solani and C. albicans keratitis decreased greatly compared with day 3, but did not change significantly in A. fumigatus keratitis. CONCLUSIONS: There is significant difference in gelatinase activities in the rabbits' corneas infected by F. solani, A. fumigatus, and C. albicans. Gelatinases play important roles in the degradation of corneal ECMs. Hyphal growth pattern and invasive depth are depended on the difference of degradations of ECMs and show difference in various fungal keratitis.

[A pharmacodynamics study of an intravitreal amphotericin B drug delivery system for the treatment of experimental Aspergillus fumigatus endophthalmitis].

OBJECTIVE: To observe a new amphotericin B drug delivery system (AmB-DDS), and investigate the therapeutic effects of AmB-DDS on an experimental Aspergillus fumigatus endophthalmitis. METHODS: (1) In order to observe the effects of AmB-DDS, thirty-four New Zealand albino rabbits were intravitreal injected Aspergillus fumigatus suspension (10(3) colony forming unit, CFU) in applanation of vitreous body before therapy 48 hours. All models were randomly divided into five groups. Group A was the empty control group, treated nothing after Aspergillus fumigatus injection, group B was the empty DDS implantation combined with vitrectomy, no treatment after DDS implanted, group C: AmB 5 microg-injection combined with vitrectomy, the injection was repeated two week later, group D: 250 microg AmB-DDS intravitreal implantation combined with vitrectomy, Group E: 500 microg AmB-DDS intravitreal implantation combined with vitrectomy. Aqueous flare, cells, anterior vitreous cells and vitreous opacity were graded, and vitreous humor smear and culture were performed at different time points after operation in 8 weeks. Two months after operation, light microscopy was used histology evaluation. (2) To observe the release of AmB-DDS in Group H (6 eyes), 500 microg AmB-DDS were implanted in the eye of the rabbits after vitrectomy, vitreous humor was aspirated and the concentrations of amphotericin B were determined by high performance liquid chromatography (HPLC). RESULTS: The inflammation response was lower in groups C, D, E than groups A, B. There was no significant statistical difference between group A and group B (P > 0.05), but differences among C, D, E and groups A, B were significant (P < or = 0.005). The inflammation grade was lower in group E than group C (P < or = 0.005). There was significant statistical difference between the cure effect of group E and group D (chi(2) = 10.494, P = 0.003). All of vitreous humor smears was positive in 1.5 months after surgery, but the culture was only positive in group A, and B. Pathological examination indicated that normal structure was disappeared in the eyes with Aspergillus endophthalmitis. At the first day after surgery, AmB were observed by analysis of HPLC, there was sustained AmB release in the group of AmB-DDS application during the observation periods. CONCLUSIONS: The degradable AmB-DDS can effectively suppress the inflammation of the rabbit model of Aspergillus fumigatus endophthalmitis. As an alternative to the current routine therapy, it can be used for the treatment of Aspergillus fumigatus endophthalmitis.

[Dexamethasone intensify the tight junctions in rat retinal vascular endothelial cells in vitro].

OBJECTIVE: To investigate the effect of dexamethasone on the expression, distribution and function of tight junctions in rat retinal vascular endothelial cells in vitro. Try to explain the mechanism of glucocorticoid in the treatment of macular edema from a new viewpoint. METHODS: Rat retinal vascular endothelial cells were isolated and purified by magnetic beads coated with anti-CD31. Cells were identified by vW factor indirect immunofluorescence staining. The fourth-passage cells were used to investigate the effect of dexamethasone on the tight junctions in rat retinal vascular endothelial cells. Cells were separated into two groups, one was treated with 500 nmol/L dexamethasone and the other was used as the control. Transepithelial electrical resistance (TER) was measured to estimate the changes in the treated group. Indirect immunofluorescent stain and RT-PCR were used to observe the difference of tight junction protein distribution and mRNA expression level between these two groups. RESULTS: Rat retinal vascular endothelial cell monolayer showed positive immunofluorescent staining for vW factor. The dexamethasone treated group showed greater TER than that of the control (P < 0.01). Tight junction protein in the dexamethasone treated group localized closer to the borders of retinal vascular endothelial cells than that of the control. Claudin-1 mRNA level of the dexamethasone treated cells were greater than that of the control. CONCLUSIONS: Dexamethasone intensifies the tight junctions in rat retinal vascular endothelial cells. Therefore this is one of the mechanisms of treatment of macular edema by glucocorticoid.