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Objective:To observe the transparency and tissue structure changes of human corneal stromal lenticules after long-term cryopreservation and explore a simple and feasible method for long-term effective preservation of corneal stromal lenticules.Methods:Two hundred samples of intact human corneal stromal lenticules from 200 eyes were obtained during femtosecond laser small-incision lenticule extraction (SMILE) in Hainan Eye Hospital, Zhongshan Ophthalmic Center from 2013 to 2020.The samples were divided into 1-month, 24-month, 60-month and 80-month group and were stored in an ultra-low temperature freezer for 1, 24, 60 and 84 months respectively at -80 ℃ according to grouping, with 50 samples in each group.Transmittance of the corneal lenticules at wavelength of 300-800 nm was measured with an ultra-micro spectrophotometer and every lenticule was measured for 10 times with a 50 nm interval.The histomorphology and collagen fiber structure of the corneal lenticules were examined by hematoxylin-eosin staining and Masson staining, respectively.The arrangement of collagen fibers and ultrastructure changes of keratocytes in the samples were inspected with a transmission electron microscope.The apoptosis rate of keratocytes was determined by TUNEL staining.The study protocol was approved by an Ethics Committee of Hainan Eye Hospital at Zhongshan Ophthalmic Center (No.2013-003). This study complied with the Declaration of Helsinki.Written informed consent was obtained from each subject before surgery.Results:The corneal lenticules were clear and intact in all groups and no significant difference in the transmittance within 450-800 nm wavelength was seen among the 4 groups (all at P>0.05). Masson staining revealed that the collagen fibers in the lenticules were neatly arranged and tightly packed in the 1-month group.In the 24-month group, interfibrous vacuoles were found in some collagen fibers.The arrangement of the collagen fibers was loose and more vacuoles were displayed in the 60-month group, and the loss of some collagen fibers appeared and the lenticules were thinned in the 84-month group.It was found through hematoxylin-eosin staining that the morphological changes of corneal stromal lenticules corresponded to the alterations of collagen fibers.Transmission electron microscopy showed that in the 1-month group, the collagen fibers of the corneal stroma lenticules were neatly arranged and regular, and the corneal stromal cells were elongated and spindle-shaped, and the nuclear membrane was intact and the cytoplasm was abundant.In the 24-month group, the collagen fibers showed slightly loose arrangement, and the corneal stromal cells were deformed with incomplete nuclear membrane.In the 60-month group, the collagen fibers were in loose and irregular arrangement, and the nuclei were atrophied and deformed.The 84-month group showed disorganized arrangement of collagen fibers, wrinkled and atrophied corneal stromal cells, discontinuous nucleus membrane and nucleoplasmic lysis.TUNEL staining showed that the percentage of apoptotic corneal cells in lenticules was (87.80±1.17)%, (89.50±1.05)%, (89.30±1.51)% and (90.20±1.47)% in the 1-month, 24-month, 60-month and 84-month groups, respectively, with no statistically significant difference found in overall comparison ( F=4.525, P=0.053). Conclusions:The disorder of collagen fibers and apoptosis of keratocytes occur in the human corneal stromal lenticules till 84 months after cryopreservation, however, the transparency and integrity remain excellent.The ultra-low temperature preservation technique provides an effective and simple solution for long-term storage of human corneal stromal lenticule.
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Objective:To investigate the in vitro interaction between antifungals and tacrolimus against Fusarium solani. Methods:According to Clinical and Laboratory Standards Institute(CLSI) M27-Ed4 and M38-A3, 24 strains of Fusarium solani isolated from clinic were used to test the minimal inhibitory concentration (MIC) of the four antifungals.Chessboard microdilution method was used to evaluate the interaction of tacrolimus (FK506) combined with four antifungals in vitro.The study protocol was approved by the Ethics Committee of Hainan Eye Hospital (No.2017-006). Results:The MIC ranges of fluconazole, voriconazole, natamycin and amphotericin B against 24 strains of Fusarium were 64-256 μg/ml, 1-8 μg/ml, 2-8 μg/ml and 1-8 μg/ml, respectively.Tacrolimus combined with fluconazole, voriconazole, natamycin and amphotericin B had synergistic effects on 12.5% (3/24), 4.2% (1/24), 16.7% (4/24) and 29.2% (7/24) Fusarium, respectively.There were no antagonistic effect in this study. Conclusions:Fusarium is sensitive to natamycin in vitro and partly sensitive to amphotericin B and voriconazole.Synergistic effects exist in few strains of Fusarium but no antagonistic effect.
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Objective:To compare the difference of the ocular surface and efficacy of orthokeratology lenses wearing after using sodium hyaluronate eye drops, rewetting drops and saline solution.Methods:A randomized single-blind controlled trial was conducted.A total of 90 right eyes from 90 myopic patients were recruited from June 2017 to April 2018 in Hainan Eye Hospital.All the subjects received vision correction by orthokeratology lens, and were randomly divided into three groups by random number table method, with 30 eyes in each group.The sodium hyaluronate group was given preservative-free sodium hyaluronate eye drops (1 g/L), the rewetting drops group was given rewetting drops, and the saline solution group was given nonpreserved 0.9% NaCl.The vision, subjective refraction, corneal topography, non-invasive first tear film break-up time(NIKf-BUT) and fluorescein stain of corneal epithelium (FL) were detected before and 1 day, 1 week, 1 month, 3 months after the lens wearing.This study adhered to the Declaration of Helsinki and was approved by the institutional Ethical Committee Review Board of Hainan Eye Hospital.Results:The NIKf-BUT in the sodium hyaluronate group at 1 day, 1 week, 1 month and 3 months after wearing orthokeratology lens was (7.13±1.71), (6.61±1.49), (6.37±1.82) and (7.01±1.63) seconds, respectively, which was obviously higher than (6.32±1.71), (5.53±1.42), (5.57±1.50) and (6.17±1.63) seconds in the rewetting drops group at the corresponding time points and (6.36±1.58), (5.81±1.49), (5.74±1.77) and (6.28±1.53) seconds in the saline solution group at the corresponding time points (all at P<0.05). There was no statistically significant difference in FL score between the rewetting drops group and saline solution group at 1 day, 1 week, 1 month and 3 months after wearing the orthokeratology lens (all at P>0.05). Lower FL scores were observed in the sodium hyaluronate group compared to the the rewetting drops group and saline solution group after wearing the orthokeratology lens (all at P<0.05). No differences were found in uncorrected visual acuity, spherical equivalent and average K value among the groups (all at P>0.05). Conclusions:Compared with the rewetting drops and saline solution, the preservative-free sodium hyaluronate eye drops can stabilize the tear film and have less effect on corneal epithelium at the early stage of orthokeratology lens wearing.However, there is no significant difference in the improvement of visual acuity and the rate of corneal shaping.
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Objective To investigate changes in the intraocular pressure(IOP) and anterior chamber angle in patients with chronic renal failure before and after hemodialysis. Methods Fifty?eight patients(116 eyes) with chronic renal failure were measured with Goldmann applanation tonometer and anterior segment optical coherence tomograph. The patients were divided into three groups based on gonioscopy results:the narrow angle group(22 eyes),the Iris neovascular group(3 eyes)and the open angle group(91 eyes). IOP was measured by Goldmann applanation tonometer in patients in the three groups before and after hemodialysis. Anterior chamber angle opening distance (AOD) was detected by anterior segment optical coherence tomograph before and after hemodialysis. The blood urea nitrogen,creatinine,albumin were also determined before and after hemodialysis. All changes in the parameters were analyzed with a paired t test. Results The IOP in narrow angle eyes and in neovascular eyes increased significantly after hemodialysis ,while the IOP in open angle eyes showed no significant changes. The AOD decreased significantly after hemodialysis. The blood urea nitrogen and creatinine decreased significantly in 58 patients,while albumin increased after hemodialysis. Conclusion The IOP in narrow angle eyes and in neovascular eyes increased after hemodialysis ,correlating with the resistance increase of aqueous outflow pathway and the change of plasma osmotic pressure. As a result ,it is recommended that the condition of eye of patients with chronic renal failure should be checked ,and patients should receive appropriate controlling measure or treatment before hemodialysis.
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Objective To observe the clinical efficacy of rigid gaspermeable contact lens (RGPCL) for correcting keratoconus and analyze the predictors for the base curve of RGPCL.Methods Data of 86 patients (143 eyes) with keratoconus fitted with RGPCL were retrospectively analyzed.All eyes were divided into the mild (n =18 eyes),moderate (n =73 eyes),advanced (n =34 eyes) and severe group (n =18 eyes).After evaluating the lens fitting,best corrected visual acuity of eyes wearing spectacle lenses and RGPCL among the 4 groups was statistically compared.And the correlation between the fitting guidance index and HL-K RGPCL base curve in each group was analyzed according to keratometry.Results All patients wearing RGPCL gained significantly better corrected vision in the mild (t =-3.336,P =0.004),moderate (t =-11.213,P ＜0.001),advanced (t =-13.959,P ＜0.001) and severe groups(t =-11.047,P ＜0.001) than those with spectacles.Equivalent spherical degree of spectacle lenses was lower than the diopter of HL-K RGPCL in the moderate (P =0.043) and severe group (P =0.006).The mean base curve of RGPCL was (50.52 ±5.64) diopter,and it was correlated with corneal curvature parameters.In mild group,the RGPCL base curve was correlated with the steep keratometry (P ＜ 0.001).The average K and relative steep K of 0.2 mm (Rm-0.2) were highly correlated with RGPCL base curve in moderate group (r =0.798,P ＜ 0.001;r =0.798,P ＜ 0.001),in advanced group (r =0.745,P ＜ 0.001;r =0.745,P ＜ 0.001) and in severe group (r =0.616,P =0.007;r =0.617,P =0.006).Conclusion Patients with keratoconus wearing HL-K RGPCL can obtain ideal corrected vision.Determination of RGPCL base curve by adjusting the corresponding indicators is conducive to improving the long-term wearing safety of the lens and enhancing the efficiency of lens fitting.
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Background Hainan Island is a tropical and subtropical area in China,with a suitable climate for fungal growth and breeding.Keratomycosis occurred frequently in this area.It is essential to analyze the causative genus and investigate the sensitive drugs prior to the clinical treatment of fungal keratitis.Objective This study attempted to analyze the fungal population of fungal corneal ulcer patients in Hainan Island of China and examine the susceptibility of antifungal drugs.Methods A series cases observational study was carried out.One hundred and seventy-four eyes of 174 fungal corneal ulcer patients were enrolled in Hainan Eye Hospital from February 2014 to May 2016 based on the diagnosis criteria in Ophthalmology.The specimens at corneal lesions were collected under the aseptic conditions,and fungus was examined by KOH based smear.Mycological culture was performed with potatodextrose agar and Sabourand's agar.The 82 cultured causative strains were identified to characterize the spectrum of the species associated with keratitis infections in Hainan Province.Susceptibilities of these causative strains to ten antifungals were determined in vitro by slip diffusion method.Results By smear microscopy,fungal hyphae and spores were found in 34 specimens of 126 patients,with the positive rate 27.0％.In 174 specimens of fungal culture,82 strains of fungi were found,with the positive rate 47.1％,and typing results revealed that the isolates were distributed among species in the fusarium (47.5％),non-spore fungus (12.2％),aspergillus (9.8％) and phoma (9.8％).Nystatin was proved to be the most effective drug to 77 strains of antifungal drug susceptibility,followed by amphotericin B and terbinafine,with the susceptibility of 88.3 ％,72.7 ％,and 71.4％,respectively.Conclusions Fusarium species,non-spore fungus,aspergillus,and phoma are common pathogenic fungi of keratomycosis around Hainan Island in China,and nystatin,amphotericin B and terbinafine are efficient in the treatment of the fungal corneal ulcer.
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<p><b>OBJECTIVE</b>To investigate the characteristics of multifocal electroretinograms (mf-ERG) of different phases in diabetic retinopathy (DR) and its clinical significance.</p><p><b>METHODS</b>Multifocal electroretinograms in patients with DR (I - II stage) were tested with VERIS IV system.</p><p><b>RESULTS</b>In I - II stage, the absolute values of N1, P1 and N2 response densities, and the N1-P1 and P1-N2 response densities were attenuated is a field of about 45 degrees in diameter.</p><p><b>CONCLUSION</b>As a new objective and quantitative examination for spatial visual function, multifocal electroretinograms may be valuable in the diagnosis of diabetic retinopathy.</p>
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Aged , Female , Humans , Male , Middle Aged , Diabetic Retinopathy , Pathology , Electroretinography , Methods , Retina , Pathology , Time FactorsABSTRACT
Objectives To investigate the expression of pax-6 in retina of infant monkeys with myopia induced by optical defocus, and to determine the role of pax-6 would play or not in onset and development of myopia and emmetropization. Methods Nine healthy infant rhesus monkeys, aged from 1 to 3 months, were selected and wore spectacle lenses or underwent photorefractive keratectomy (PRK).Transcription polymerase chain reaction method and quantitative analysis were used to determine the expression of pax-6 in the retina with myopia induced by optical defocus in different time, and the result was compared with that in retina without myopia. Results The myopia caused by hyperopic defocus was found. The expression of pax-6 in the retina with myopia induced by optical defocus was significantly higher than that in the retina without myopia(t=3.480,P=0.004). Conclusions The expression of pax-6 is enhanced by hyperopic defocus in the infant monkey retina, which suggests that pax-6 may be involved in vision-dependent eye growth and emmetropization.