ABSTRACT
Background Blepharitis is a common ocular surface disease.It is associated with the disorder of lipid secretion of meibomian gland.The change of tear film stability can cause dry eye symptoms,so blepharitis is thought to be one of the factors causing dry eye,but the relation between them is in study. Objective This study was to observe the morphology of meibomian gland in blepharitis patients and to investigate the correlation of morphology of meibomian gland with dry eye. Methods A series of case-observational study was designed in this study.A total of 83 eyes of consecutive 83 blepharitis patients were enrolled in Henan Eye Institute from October 2010 to April 2011.Blepharitis was diagnosed based on American Preferred Practice Pattern Guidelines.Some relevant ocular examinations were performed under the informed consent of the subjects,including the anterior segment manifestation by the slit lamp,such as meibography,lid margin abnormality,and the dry eye-relevant examinations,such as tear film break-up time (BUT),Schirmer test Ⅰ and corneal fluorescein staining also been carried out.Tear film shape was examined by film interference images and scored.Absent degree of meibomian gland was graded under a Noncontact Infrared Meibography.The correlations of absent degree of meibomian gland with ocular syndrome score,dry eye examination results were evaluated using Spearman rank correlation coefficients.Informed consent was obtained prior to this trail. Results No significant difference in the frequencies of blepharitis was found between male and female among different ages (x2 =2.69,P =0.75 ).Absent grading of the meibomian glands was positively correlated with age of blepharitis patients ( r =0.58,P =0.00 ),lid margin abnormality scores ( r =0.64,P =0.00 ),conjuntival hyperemia score ( r=0.50,P =0.00),tear film interference imaging grade ( r =0.23,P =0.04 ),corneal fluorescein staining score( r =0.50,P =0.00 ) but was negatively correlated with BUT ( r =-0.32,P =0.00 ).No significant correlation was found between meibography grading and gender( r =-0.09 ; P =0.99 ) or Schirmer test Ⅰ ( r =-0.05;P =0.69 ).No significant difference was found in meibography grading between male and female in different age groups(Z=-0.09,P=0.93). Conclusions Blepharitis can irriter dry eye symptom because of overevaporation of tear fluid and abnormality of secreting function of meibomian glands.The missing of the meibomain glands increases with age in the patients with blepharitis.Noncontact Meibography System is an assistant tool to the diagnosis of blepharitis.
ABSTRACT
Background Infective keratopathy is a key cause of corneal blindness in China,and fungal keratitis is proved to have a higher incidence and bigger threats in infective keratitis.Researches showed that topical immunology plays an important effect during the development of fungal keratitis,but its mechanism is still studying.Objective This experiment was to explore the critical immunocyte during the process of fungal keratitis.Methods Forty-eight SPF 12-week-old male C57BL/6J mice were included and randomized into the control group and model group.The fungal keratitis model closely mimicking human cornea infections was established in the mouse using scratch followed by incubation of fusarium solani on the cornea,and the mice in the control group scratched on the cornea only.Cornea was examined under the slit lamp at 0,6,9,12,24,72 and 120 hours after operation.The severity of keratomycosis was clinically scored based on the literature criteria.The inflammatory cells were identified using immnofluorescence label,and the number of the inflammatory cells was calculated and compared among different groups and time points.This study complied with the Statement of ARVO in the use of experimental animal.Both Experimental Animal Ethic Commission in Zhengzhou University and Life Science Management Commission approved this study proposal.Results After inoculation of fusarium solani,typical fungul keratitis signs were seen on the cornea.Severe corneal opacifieation occurred within 24 hours and peaked at 72 hours.However,only mild edema of cornea was exhibited and gradually recovered normal in the control group within 24 hours.The clinical score of inflammation was higher in the model group in various time points than that in the control group,and it was seen that 24-72 hours after operation,the score attached peak in the model group with a significant difference in comparison with the control group(P＜0.01).In 9,12,24,72 and 120 hours after operation,the number of neutrophil cells was significantly increased in the model group compared with control group (P＜0.05),and that in 12,24,72 hours after operation was significantly higher than the 6 hours(P=0.004,0.000,0.001).However,no significant differences were seen in the number of neutrophil cells between 9 or 120 hours and 6 hours after operation(P=0.772,0.323).The number of T lymphocytes in cornea was significantly increased in 72 and 120 hours in comparison with 6 hours in the model group(P=0.000,0.000),and from 72 to 120 hours after operation,the number of T lymphocytes was significantly higher than that of the contral group (P＜0.01).The neutrophil cell number was positive correlated with the inflammatory score in the early phase (r =0.593,P =0.000).T limphocyte emerged in late phase but no significant correlation with the clinical score (r＝0.315,P=0.062).Conclusions Neutrophil cells play a critical role in the development of fungal keratitis in early stage.
ABSTRACT
Background Human limbal allograft transplantation or limbal autograft transplantation are the primary approaches to the severe corneal-blindness,but their application in clinic were limited because of the defects of donor material.With the development of tissue engineering technology,transplantation of in vitro cultured limbal epithelial stem cells is being an advanced management.Objective The aim of this work was to expand human limbal epithelial stem cells ex vivo under the guidance of confocal microscope and to lay the foundation for fabricating ex vivo cultured cell sheets.Methods Ten eyes of ten patients were examined with the Heidelberg Retina Tomography Ⅲ Rostock Cornea Module(HRT3-RCM)to elucidate the structure of the human corneoscleral limbus and to correlate limbal epithelial dimensions.According to the analysis of the images of limbal epithelia,the limbal tissues provided by Eye Bank of Henan Eye Institute were cut into suitable explants.Then,this study was conducted to expand limbal epithelial stem cells ex vivo on denuded amniotic membrane.The phenotypes of primary cultured cells were evaluated by morphology and immunofluorescent staining with antibodies for limbal epithelial stem cell markers (p63,cytokeratinl9)and differentiation markers(keratin 3,involucrin).This experimental procedure was approved by the Ethic Committee of Henan Provincial People's Hospital.The written informed consent was obtained from subjects before initiation of any examination.Results The palisade morphology of human limbus was imaged clearly on the laser scanning in vivo confocal microscopy and many hyperreflective cells were observed in palisade basal cells.The cell-island phenomenon was seen in the basement membrane under the laser scanning in vivo confocal microscopy.The oblique sections of limbus showed many papilla-like epithelial columns below the superficial limbal epithelia.Throughout the experiment duration,the epithelial cells grew well with the migration rates from limbal tissue (68.62± 16.94)％ and the migration time(5.83 ±2.04)days,which depended on the tissue freshness.Compared with the second and forth batch of tissue,the migration rates of the third and sixth batch of tissues were significantly higher(P＜0.05),and the migration time was evidently longer in the forth and sixth batch of tissue compared with the first,second,third and fifth batch(P＜0.05).The positively expressing rates in the cultured corneal stem cells were 4.05％ and 36.52％ for p63,26.07％ and 40.55％ for CK19,57.88％ and 40.81％ for K3,64.66％ and 59.19％ for involucrin.Conclusion Human limbal epithelial stem cells can be successfully and purposefully obtained from the limbal tissue based on the guidance confocal miscroscope.The cultured corneal stem cells can grow well on the denuded amniotic membrane