To investigate the efficiency of the transfection of PEGFP-IL-1ra plasmid via cation polymer mediation (poly-ethylenimine, PEI) by injection into the corneal stroma.Plasmid PEGFP-hIL-1ra recombinants were constructed and transferred into corneal endothelial cells via cation polymer mediation. Plasmid PEGFP-hIL-1ra recombinants and/or PEI-in-vivo were injected into the corneal stroma of Wistar rats. Corneas were harvested at different time points (days 3, 6, 14 and 21) after injection. The expression of IL-1ra after transfection was studied by fluorescence microscopy, transmission electron microscopy, real time polymerase chain reaction (RT-PCR) and immunohistochemistry.Plasmid PEGFP-hIL-1ra recombinants were constructed successfully. After injection of pEGFP-hIL-1ra plasmid into the cornea, IL-1ra mRNA expression was detected in the corneal stroma and reached a peak on day 6. IL-1ra-GFP granules could be observed in every layer of the cornea in the PEGFP-hIL-1ra recombinant group by transmission electron microscopy, but not in the negative control (PEI-in-vivo) group. P63 immunocytochemical staining in the corneal epithelium was positive in both groups. There was no impairment in the ultrastructure of cells in both groups.By direct injection of PEGFP-hIL-1ra into the corneal stroma and mediation by the cation polymer, the IL-1ra gene could be transferred and expressed in corneal tissue efficiently. This may be a novel technique for gene transfection to the cornea in situ.
It is reported that retinal abnormities are related to Alzheimer's disease (AD) in patients and animal models. However, it is unclear whether the retinal abnormities appear in the mouse model of sporadic Alzheimer's disease (sAD) induced by acrolein. We investigated the alterations of retinal function and structure, the levels of β-amyloid (Aβ) and phosphorylated Tau (p-Tau) in the retina, and the changes in the retinal vascular system in this mouse model. We demonstrated that the levels of Aβ and p-Tau were increased in the retinas of mice from the acrolein groups. Subsequently, a decreased amplitudes of b-waves in the scotopic and photopic electroretinogram (ERG), decreased thicknesses of the retinal nerve fiber layer (RNFL) in the retina, and slight retinal venous beading were found in the mice induced by acrolein. We propose that sAD mice induced by acrolein showed abnormalities in the retina, which may provide a valuable reference for the study of the retina in sAD.
Thirteen new Euphorbia diterpenoids, euphylonanes A–M (1–13), and eight known ones were isolated from the whole plants of Euphorbia hylonoma. Compounds 1 and 2 are two rearranged ingenanes bearing a rare 6/6/7/3-fused ring system. Compound 3 represents the first example of a 9,10-epoxy tigliane, while 4–21 are typical ingenanes varying with substituents. Structures were elucidated using a combination of spectroscopic, computational, and chemical methods. Most ingenanes exerted a significant antiadipogenic effect in 3T3-L1 adipocytes, among which 4 was the most active with an EC50 value of 0.60 ± 0.27 μM. Mechanistic study revealed that 4 inhibited the adipogenesis and lipogenesis in adipocytes via activation of the AMPK signaling pathway.
Objective To explore the indications, efficacy, and complications from different types of lamellar keratoplasty (LKP) for Mooren's corneal ulcers.Methods It was a retrospective case series study.Twenty-six patients (31 eyes) with Mooren's corneal ulcers underwent crescent LKP (14 eyes),D-shaped LKP (7 eyes),ring-shaped LKP (3 eyes),or total LKP (7 eyes) at Zhongshan Ophthalmic Center between January 2008 and June 2011.The selection of surgical procedures was based on the size and shape of the corneal ulceration.Best corrected visual acuity (BCVA),corneal graft clarity, astigmatism, and complications after the operation were studied during the follow-up period of 6-36 months.Results Patients with the BCVA between 0.3-0.5 after operation including,crescent LKP (3 eyes),D-shaped LKP (4 eyes),total LKP (5 eyes).Patients with the BCVA between 0.6-1.0 after operation including,crescent LKP (ll eyes),D-shaped LKP (3 eyes),ring-shaped LKP (3 eyes),total LKP (5 eyes).Visual acuity in 17eyes (54％) was improved after LKP,12 eyes (40％)remained the same,while 2 eyes (6％) decreased after operation.The mean corneal astigmatism in the ring-shaped graft group was 2.30±0.40 D,and 3.70±1.03 D in the D-shaped graft group 6 months after the operation.Postoperative complications included corneal epithelium defect in 4 eyes (13％),recurrence of Mooren's ulcer in 2 eyes (6％),suture-related corneal abscess in 1 eye (3％),and graft rejection in 4 eyes (13％).Conclusion LKP is effective in restoring the integrity of eyes with Mooren'sulcer.Designing the appropriate type of LKP based on the different shapes of corneal ulceration allows the removal of the corneal lesion and spares the maximum amount of normal tissue.Furthermore,a well-matched graft configuration can reduce surgically related astigmatism, and help to improve vision in most cases.
Key words:
Mooren's ulcer; Lamellar keratoplasty; Postoperative complication; Astigmatism
To investigate the possibility of ocular surface reconstruction with amniotic membrane in the acute stage of burn injury, to compare the results using fresh and preserved amniotic membranes and to evaluate the surgical methods and their effects.Consecutive patients of whole corneal burn above degree III with complete destruction of the limbus were divided into two groups to receive amniotic membrane transplantation (8 eyes of 8 patients with fresh amnion, 12 eyes of 11 patients with preserved one) or lamellar keratoplasty (24 eyes of 22 patients). The follow-up period was 12 to 26 months with an average of (15 +/- 2) months.The ocular surface became stabilized after the transplantation of amniotic membrane. In eyes treated with fresh amniotic membrane, the corneal surface was epithelized immediately. In eyes treated with preserved amniotic membrane, the corneal surface was epithelized only after 2 - 3 weeks. Lamellar keratoplasty was performed in 3 of 12 eyes with preserved amnion transplantation because the amnion was dissolved due to persistent epithelial defects. Amnions were absorbed with time and superficial neovascularization followed in the corneal surfaces. Seven of eight transplanted fresh amnions were absorbed within 2 to 8 months [mean time (4.3 +/- 0.8) months], and 10 of 12 preserved amnions were absorbed within 1 to 3 months [mean time (2.0 +/- 0.3) months]. The difference was statistically significant (t = 4.22, P < 0.01). The conjunctival surface was successfully reconstructed with amniotic membrane at the acute stage of burn injury. Moderate symblepharon occurred in one case only. Corneal dissolution never occurred in all patients who received lamellar keratoplasty, but recurrent erosion of corneal epithelium occurred in the grafts and corneal neovasularization developed eventually. Corneal graft had to be performed again on four eyes and symblapharonplasty had to be performed on seven eyes. Mild to medium symblepharon was observed in 5 of 24 eyes received lamellar keratoplasty. Visual acuity could be maintained at hand movement in eyes treated with amniotic membrane transplantation without secondary glaucoma and cataract. Visual acuity was figure counting in most cases with lamellar keratoplasty.Amniotic membrane transplantation (especially using a fresh membrane) can effectively reduce the inflammation of the cornea at the acute stage of burn injury, can prevent corneal ulcer and perforation and can make the stabilization process faster. It can also decrease corneal neovascularization as well as establish better conditions for successful keratoplasty.
To investigate the stability of FK506 eye suspension and its pharmacokinetics in rabbit aqueous humor, as well as its distribution in eye tissues.Sedimentation rate, flocculation value, redispersion time, rheological study, and accelerated experiment were determined for evaluating the stability of FK506 suspension. In a single-dose pharmacokinetic study, six rabbits were instilled a 25-microL drop of 0.05% FK506 suspension and aqueous humor samples were collected at different intervals after administration. In a multiple-dose pharmacokinetic study, a 25-microL drop of FK506 suspension was instilled into the right eye of six rabbits four times a day for 7 days. On the eighth day, aqueous humor samples were collected before the administration of the first, second, third dose, and at different checkpoints after the third dose. For tissue distribution study, six eyes per time points (18 rabbits in total) were treated with single dose of FK506 suspension, and the eyes were enucleated at 60, 100, and 240 min after treatment, then eye tissues were collected. The concentrations of FK506 in all samples were determined by LC-MS/MS.The preliminary results indicated that the stability of FK506 suspension was in accord with the standards of Chinese pharmacopoeia. The maximum concentrations of aqueous humor after single dose and multiple dose administrations were 31.40 +/- 9.32 ng/mL and 37.73 +/- 11.25 ng/mL, respectively. The concentration of FK506 in cornea at 60, 100, and 240 min after a single dose were 402.0 +/- 96.8 ng/g, 363.8 +/- 84.5 ng/g, and 220 +/- 62.3 ng/g, respectively. Determination of pharmacokinetic parameters of single-dose and multiple-dose administration, as well as the FK506 concentrations in eye tissues, showed that the FK506 formulation and the dosing regimen ensured the therapeutic concentration of FK506 for treating corneal allograft rejection.Based on the stability, single-dose and multiple-dose pharmacokinetics, and tissue distribution, FK506 suspension eyedrops may be a suitable candidate for clinical application in ophthalmology.
Herpes simplex keratitis (HSK) is a recurrent inflammatory disease of cornea primarily initiated by type I herpes simplex virus infection of corneal epithelium. However, early diagnosis of HSK remains challenging due to the lack of specific biomarkers. This study aims to identify biomarkers for HSK through tear metabolomics analysis between HSK and healthy individuals. We conducted a cross-sectional study enrolling 33 participants. Tear samples were collected from one eye of 18 HSK patients and 15 healthy volunteers using Schirmer-strips. Tear metabolomic profiling was performed using high-performance liquid chromatography tandem mass spectrometry (LC–MS/MS). Metabolites were quantified and matched against entries in the human metabolome database (HMDB) and small molecule pathway database (SMPDB) to identify metabolites and metabolic pathways, respectively. Metabolic differences between HSK and control group were determined using multivariate statistical analysis. A total of 329 metabolites were identified, of which 18 were significantly altered in HSK patients. Notably, 12 metabolites were significantly increased, and 6 were significantly decreased in HSK patients. The changed metabolites were enriched in these pathways: arginine and proline metabolism, phospholipid biosynthesis, alpha linolenic acid and linoleic acid metabolism, retinol metabolism. To assess the potential utility of tear biomarkers, a predictive model was developed combining 4 metabolites (AUC = 0.998 [95%CI: 0.975, 1]): D-proline, linoelaidic acid, plantagonine, and phosphorylcholine. Our study establishes that HSK has a distinctive metabolomic profile, with 4 key elements maybe emerging as potential biomarkers for diagnostic purposes. These findings may provide novel insights into early and rapid diagnosis of HSK.
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