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Determination of coenzyme Q10 level in human retina by the high-performance liquid chromatography

Jinfeng, QU ; Xiaoxin, LI ; Washington ILYAS

Chinese Ophthalmic Research.2010;28(3):253-256. doi:10.3969/j.issn.1003-0808.2010.03.017

Background Researches have demonstrated that age-related macular degeneration (AMD) is associated with the oxidative stress injury of retina.Coenzyme Q10 (CoQ10) is an important antioxidant agent.CoQ10 level in blood plasma is a primary index of reflecting the oxidative stress ability of human.However,the study on CoQ10 content in retina has not been seen yet.ObjectiveThe aim of this study is to establish a method of detecting CoQ10 content in retina by the high-performance liquid chromatography (HPLC). Methods The retinas were isolated from 10 healthy eyes of donors aged 20-28 years.The donor eyes were obtained from National Development and Research Institute,Inc.USA.Isolated retina tissue was prepared into homogenate then lyophilized and deproteinized with methanol.Samples were extracted with heptane prior to the HPLC analysis with the chromatographic conditions as follows:RP-18 column,a mobile phase consisted of methanol-hexane-acetic acid-isopropanol (V/V=55:9:1:1) and 0.42% sodium acetate,ultraviolet rays (UV) detector at 275 nm.Results CoQ10 was effectively isolated from human retina.The limit of detection of CoQ10 was 0.14mg/L.The peak area and concentration of CoQ10 showed a good linear correlation within the concentration range of 0.2-395.00mg/L (R~2＝0.9943).Repeatability study showed that the relative standard deviations for CoQ10 at the concentration of 0.86mg/L,2.59mg/L and 3.45mg/L were 2.7%,0.1% and 3.3%,respectively.The within- and inter-day standard deviations for the analysis of CoQ10 were 1.6% and 3.7%,respectively.The recovery was 101%-113% for the human retina samples.The concentration of CoQ10 in 10 retinas from human donors was 0.51±0.20μg/eye in average.Conclusion A HPLC method for the quantified analysis of CoQ10 in human retina is developed.

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The development of the applications of ocular response analyzer in excimer laser surgery

Ying, YANG ; Xingtao, ZHOU

Chinese Ophthalmic Research.2010;28(3):285-288. doi:10.3969/j.issn.1003-0808.2010.03.026

Research on the application of ocular response analyzer has become one of the main issues in ophthalmology.The ocular response analyzer has been used in the measurement of ocular bioparameters of glaucoma,cataract,heretability and refractive surgery.As a newly invented non-contact tonometric device,the ocular response analyzer is helpful not only in the measurement of intraocular pressure (IOP),but also in the determination and calculation of two corneal biomechanical properties:corneal hysteresis (CH) and corneal resistance factor (CRF).It offers the possibility of measuring corneal biomechanical properties in vivo.This article primarily reviewed the application of the ocular response analyzer in terms of pre-operative examination and post-operative follow-up in excimer laser surgery.

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The anterior chamber depth measurement of primary angle-closure glaucoma with UBM and pentacam

Liang, JUAN ; Liu, WEI ; Ji, JIAN

Chinese Ophthalmic Research.2010;28(1):75-78. doi:10.3969/j.issn.1003-0808.2010.01.020

Background Nowadays,glaucoma is the major cause of irreversible blindness worldwide and the second cause of global visual loss.To accurately obtain the anterior chamber depth (ACD)is of helpful for screening primary angle-closure glaucoma (PACG).ObjectiveThe aim of this study is to explore the difference and agreement in the measurement of ACD between Scheimplug Imaging System (Pentacam)and ultra-sonographic biomicroscope (UBM).MethodsSeventy-seven eyes of 77 PACG patients aged (69.96±7.87)were divided into two groups.Thirty-seven eyes with acute primary angle-closure glaucoma (APACG)were assigned to group 1,and forty eyes with chronic primary angle-closure glaucoma (CPACG)were assigned to group 2.Central ACD was measured with Pentacam and UBM,respectively.The Bland-Altman method was used to evaluate the agreement of the two methods.ResultsThe ACD values measured by Pentacam and UBM were (1.6467±0.2687)mm and (1.5601±0.2677)mm respectively in APACG,indicating a significant difference (t=-7.259,P<0.01)and a positive linear correlation between the results of these two methods (r=0.939,P<0.01).The ACD values measured by Pentacam and UBM were (2.0622±0.2317)mm and (1.9648±0.2176)mm respectively in CPACG eyes with a significant difference between them(t=-10.433,P<0.01)and a significant linear correlation between these two outcomes (r=0.967,P<0.01).The Bland-Altman method showed that the two modalities had comparable results for ACD.ConclusionPentacam,as a new 3-dimensional mathematical model of the anterior segment,presents some different results from UBM in the measurement of ACD,but it is not clinically significant.Combination of Pentacam and UBM may be available for the clinical measurement of ACD.

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Study on association of the central cornea histology change with age in adults by in vivo confocal microscopy

Liping, QIAO ; Huimin, SUN ; Shaozhen ZHAO ; Jiaqin, YUAN

Chinese Ophthalmic Research.2010;28(1):66-70. doi:10.3969/j.issn.1003-0808.2010.01.018

Background Corneal confocal microscopy can denamically offer the morphological image of corneal cells at cellular level.To find out the dynamic change of corneal thickness and morphology of corneal cells with aging is very important for the design of corneal refractive surgery.ObjectivePresent study is to find out the important relevance of dynamic change of corneal thickness and morphology of corneal cells with aging and the design of corneal refractive surgery and investigate the influence of age on central corneal tissue and characterize precisely the anatomy of cornea by in vivo confocal microscopy.Methods122 eyes of 122 normal subjects were enrolled in this study with the age from 18 through 80 years old.The subjects were diagnosed as emmetropia in the Center of Tianjin Medical University from August 2003 to December 2007.All eyes were examined in vivo by confocal microscopy.The cell morphology,cell density and corneal thickness were measured by confocal microscopy.The relationship of central corneal tissue change and age was evaluated.ResultsThe density of corneal superficial basement epithelial cells showed a significantly negative correlation with age (r=-0.355,P=0.017).The keratocyte density in the anterior and posterior stroma indicated a significantly negative correlation with age (r_1=-0.462,P=0.001;r_2=-0.403,P=0.016).The thickness of corneal epithelium cells had a significantly negative correlation with age (P=0.02).The mean value of total corneal thickness and stromal thickness in high myopic eyes was (523.2±26.20)μm and (468.4±20.72)μm respectively,presenting a significantly negative correlation with age (r_1=-0.552,P=0.014;r_2=-0.545,P=0.035).A significantly negative correlation was also found in endothelial cell areas and density with age (r_1=0.417,P=0.004;r_2=-0.598,P=0.002).The percentage of polymorphological corneal endothelial cells was considerably increased(r=0.417,P=0.004)but that of pleomorphism cells was significantly decreased in >60-year-old population compared with younger subjects (r=-0.598,P=0.002).The morphology of corneal basement cell,anterior stroma cytocyte and endothelial cell were normal in younger subjects,and enlargement of the cells in size,decreases of number and density of the cells were found in >60-year-old subject.ConclusionThe anatomy and histology of the central cornea in young subjects exist discrepancy from older ones.The corneal hisological changes with aging include thinner thickness of epithelium,thinner thickness of cornea,decreased density of keratocyte and endothelial cells and increased size of endothelial cells.Corneal thickness is asymmetric and seems to undergo age-related anatomic changes.

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Clinical application of superficial temporal artery island flap in the reconstruction of eyelid and periorbital defect

Zhengkang LI ; Xianqun, FAN

Chinese Ophthalmic Research.2010;28(1):55-57. doi:10.3969/j.issn.1003-0808.2010.01.015

Serious defect of eyelid and periorbital tissue affects the appearance of patients and closure-eye function.Application of superficial temporal artery island flap to repair the defected tissue is considered to be an available method to reconstruct eyelid and periorbital tissue.ObjectiveThis study aims to investigate the indication,safety and effectivity of reconstructing the serious defect of eyelid and periorbital tissue by forward flow flap.MethodsEighteen eyes of 18 cases with serious eyelid and periorbital tissue defect were reconstructed by covering the defect zone with superficial temporal artery to form forward flow flap.The flap was made along the distribution of superficial temporal artery with the size over the tissue detect area and transferred to lesion via subcutaneous tunnel.The follow-up time was 6 months.Survival status of flap,plastic condition of lesion and surgical treatment were discussed.ResultsAmong 18 cases with eyelid and periorbital defect,the flaps were survived in 16 cases with the satisfactory repairing appearance,showing nice color,soft texture and normal sensation after plastic surgery.Reconstructed eyelids closed well.One case showed the temporary obstruction of venous refluence in early stage after operation and remained distal epidermal necrosis.Reoperation was performed in 6 patients because of flap hypertrophy and showed a successful outcome.One patients occurred the failure of plastic flap due to hypertrophy and underwent free-flap transplantation in 3 months following the first surgery and obtained a ultimately flap survival.ConclusionThe superficial temporal artery forward flow flap offers a stable and reliable blood supplying and provides adequate tissue of excellent color and texture match.It is an ideal and available flap for the reconstruction of serious eyelid and periorbital tissue defect.

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Evaluation of the relationship between donor graft position and recipient position after endothelium keratoplasty by AS-OCT

Xin, WANG ; Jing, HONG ; Feng, LIU ; Yanfang, YANG ; Aihua, DING ; Jing, REN ; Danyang, ZAN

Chinese Ophthalmic Research.2010;28(1):50-54. doi:10.3969/j.issn.1003-0808.2010.01.013

Background The advantages of endothelium keratoplasty(EK)include without corneal surface incision and sutures,remaining normal corneal topography,minimizing astigmatism and providing a healthy donor endothelial cell count and function.Using anterior segment optical coherence tomography(AS-OCT) to examine the morphology of the corneal graft after EK is helpful for the evaluation of the outcome of EK.ObjectiveThis study applies AS-OCT to observe the relationship of recipient cornea and donor graft and offer a basis for the evaluation of operative outcome.MethodsThe clinical data from 44 eyes of 42 patient who received DSAEK in Peking University Eye Center from September 2007 through April 2009 was reviewed retrospectively.These patients included 30 male and 12 female with the age from 15 to 83 years.Structure of cornea and relationship between recipient cornea and graft disc after surgery were clinically examined and recorded by slit-lamp digital camera and AS-OCT.This study was approved by the Ethics Committee of Peking University.Written informed consent was obtained from all of the patients before the operation.ResultsThe donor graft attached to posterior central surface of recipient in 28 eyes under the slit-lamp microscopy and AS-OCT.The thickness of donor graft was decreased,and graft sticked more closely to recipient cornea and was more transparent with the lapse of time.Complete dislocation of the donor graft in early stage was found in 5 eyes(11.36%) and partial dislocation in 2 eyes(4.55%).Curliness of graft edge occurred in 1 eye(2.27%) and eccentricity of donor graft in 8 eyes(18.18%).ConclusionAS-OCT can display the morphology and structure of cornea as well as the relationship between donor graft and recipient after DSAEK.It is helpful for evaluating the clinical effectiveness of DSAEK.

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Expression of AQP-1 in rat cornea neovascularization after alkali burn

Zhi, HE ; Mingchang ZHANG ; Ying, ZHANG

Chinese Ophthalmic Research.2010;28(1):25-28. doi:10.3969/j.issn.1003-0808.2010.01.008

Background Corneal alkali burn is a common cause of corneal neovascularization(CNV).Researches demonstrated that aquaporin 1(AQP1) plays key role in neovascularization of tumor tissue.So it is speculated that AQP1 participates in pathogenesis and development of CNV.ObjectivePresent study aims to investigate the expression of aquaporin 1(AQP1) in cornea after alkali burn and explore its role in corneal neovascularization.MethodsCornea neovascularization (CNV) models were established by putting a filter paper (the size of 3 mm in diameter immersed 1mol/L NaOH solution) on cornea for 20 seconds in the left eyes of 25 adult Sprague Dawley rats.The fellow eyes were treated using the filter paper with normal saline solution as normal control group.Corneal new blood vessel was evaluated under the slim lamp biomicroscopy once per day,and area of new vessel was calculated.The animals were sacrificed and corneal samples were obtained in day 1,4,7,14,21 after corneal alkali burn.The expression of AQP1 and VEGF in corneas were detected by immunochemistry,and the relative expression level of AQP1 mRNA in rat cornea was determined using RT-PCR.ResultsThe corneal neovascularization grew at the second day and reached the largest area at the fourteenth day after corneal alkali burn.The expression of VEGF protein (gray value) in corneal sample was significantly elevated from the first day through the fourteenth day after corneal alkali burn in comparison with control group(t_(1d)=12.410,t_(4d)=29.428,t_(7d)=16.030,t_(14d)=13.321,P＜0.05),and then the expression level declined till twenty-one day without obvious difference in comparison with control group(t_(21d)=1.587,P＞0.05).The expression of AQP1 protein showed the same tendency with that of VEGF protein(t_(1d)=7.623,t_(4d)=15.293,t_(7d)=18.291,t_(14d)=14.483,t_(21d)=6.381,P＜0.05).The dynamic change expressions of AQP1 and VEGF showed a significant positive correlation(r=0.834,P＜0.05).The expression of AQP1 mRNA (A value)was weaker in control group but considerably increased after alkali burn with the peak value in the seventh day,showing a significantly difference between day 1,4,7,14 and control group(t_(1d)=3.491,t_(4d)=10.690,t_(7d)=12.936,t_(14d)=10.767,t_(21d)=8.594,P＜0.05).ConclusionThe level of AQP1 expression is markedly related to inflammatory CNV in rat cornea after alkali burn.

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The influence of negative suction during LASIK on content of retinal amino acids in rabbit

Haixia, ZHAO ; Zhiying, GUO ; Chunmei, NIU ; Wenying, GUAN

Chinese Ophthalmic Research.2010;28(1):5-9. doi:10.3969/j.issn.1003-0808.2010.01.002

Background Whether negative suction during excimer laser in situ keratomileusis(LASIK) affect the structure and function of retina or not is in controversy,but it seems that temporary hypertention induced by negative suction is a key factor of impairment of retina in LASIK.ObjectiveThis study attempts to study the influence of transient high intraocular pressure (IOP) during LASIK on the contents of retinal amino acid.MethodsThe both eyes of 45 New Zealand white rabbits were suctioned for different periods (20s,45s,3min) with negative pressure generator during the LASIK to make the instantaneous high IOP models,and LASIK without negative suction was performed in the both eyes of 15 rabbits in the control group.The changes of the contents of retinal amino acids were evaluated with High Performance Liquid Chromatography(HPLC) at 0,7,10,14 and 28 days postoperatively and compared with those of control group.ResultsThere were no statistically significant differences in the contents of retinal amino acids among different time points after operation in negative suction for 20s group and 45s group,respectively(P＞0.05).At negative suction for 3minuts,the content of glutamic acid in retina was significantly increased in comparison with control group in 7,10,14 and 28 days (P＜0.05),and no statistically significant difference was seen in the contents of glutamic acid at postoperative instant group compared with control group(P＞0.05).A statistically significant difference in the contents of glutamine,tryptophan,phenylalanine was revealed among postoperative 10 days,14 days and 28 days groups comparison with control group (P＜0.05).ConclusionThe acute IOP elevation caused by negative suction during LASIK results in the reversible increase of retinal amino acids.The duration of negative suction time influent the reconstruction of retinal structure.

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Application of ultrasound biomicroscopy in open-globe injury

Chaojun, WANG ; Lingjie, ZHANG ; Qinhui, XIN

Chinese Ophthalmic Research.2010;28(3):278-280. doi:10.3969/j.issn.1003-0808.2010.03.024

Background Open-globe injury of zone Ⅱ is a severe eyeball wound in the sclera area near 5mm from the corneal limbus.To accurately evaluate the local tissue structure after the wounding is very important for lessening the complications of eye injury.Ultrasound biomicroscope (UBM) has been proven useful in examining eyeball wall and vitreous at the wound in order to offer the useful information about whether secondary operation is needed for the injured eyes and when is the optimal timing for surgery.Objective The present study is to explore the clinical application of UBM in open-globe injury of zone Ⅱ. Methods This is an observational study of clinical cases.UBM examination was performed on 38 eyes of 38 cases received emergency repair surgery of open-scleral injury.The restore status of sclera,ciliary body and vitreous of injuried zone (zone Ⅱ) were evaluated under the UBM in 3-5 days after the operation.The comprehensive results of UBM,B-scan and ophthalmoscope examinations were analyzed to determine the further treatment.The oral informed consent was obtained from each subject before this trial.Results Vitreous surgery were performed on 15 cases,and all wounds were found by UBM and were proven in the surgery.The other 23 cases were followed up in out-patient department and no obvious complications were found.Conclusion UBM is a valuable and atraumatic examination for open-globe injury of zone Ⅱ.It offers important and appraisal information about ocular injury.

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Measurement of optic disc parameter of Chinese normal people with Heideberg retina tomography Ⅱ

Jing, WANG ; Chenming ZHANG ; Bailing, GUO

Chinese Ophthalmic Research.2010;28(3):275-277. doi:10.3969/j.issn.1003-0808.2010.03.023

Background Heideberg retina tomography Ⅱ(HRTⅡ) can offer the quantitative description of optic disc topography and optic nerve fiber layer thickness.The normal optic disc parameter by HRT Ⅱ from Chinese has been reported,but the comparisons between males and females or the left and right eyes are lack.ObjectiveThe goal of this study is to measure the optic disc topography of healthy people with HRT-Ⅱ.Methods Four hundreds eyes from 108 healthy male subjects and 92 healthy female subjects aged 10-72 years were measured with HRT Ⅱ in this study.The parameters of optic disc including disc area,cup area,rim area,cup/disc area ratio,rim/disc area ratio,cup volume,rim volume,cup depth,maximal cup depth,height variation,mean RNFL thickness,and RNFL cross-sectional area were analyzed and compared between the gender or left and right eyes.Oral informed consent was obtained from all of the subjects before this procedure.Results There were no significant differences in measuring parameters by HRTⅡ between female and male subjects (P＞0.05) or between right eye and left eye (P＞0.05).Conclusion The results can be used as a normal reference of optic disc parameters of HRT Ⅱ.

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