INTRODUCTIONThe proteins found in tears play an important role in maintaining the ocular surface and changes in tear protein components may reflect changes in the health of the ocular surface. Proteomics provides a comprehensive approach for cataloguing all the proteins of the tear proteome, which will help to elucidate disease pathogenesis, make clinical diagnoses and evaluate the influence of medications on the structure, composition and secretion of tear proteins. In this study, an alternative proteomic strategy was investigated to explore the human tear proteome.

MATERIALS AND METHODSTear samples were obtained from patients who had pterygium and were collected on the first day and third day after pterygium surgery. Tears pooled from 6 patients were used in the analysis. Reverse-phase high-pressure liquid chromatograph (RPHPLC) was used as the first step to separate intact proteins into 21 peaks. Each fraction was then tryptic-digested and analysed by nanoLC-nano-ESI-MS/MS to characterise the protein components in each fraction.

RESULTSIn total, 60 tear proteins were identified with high confidence, including well-known abundant tear proteins, and tear-specific proteins such as lacritin and proline-rich proteins. Among them, proline-rich protein 5 was found for the first time in tear fluid. A large number of plasma proteins were also observed in tear fluid.

CONCLUSIONSThe results showed that the proteomic strategy used in this study was successfully applied to analyse tear proteome.

Eye Proteins ; analysis ; Humans ; Mass Spectrometry ; methods ; Proteome ; Tears ; chemistry

Measuring the tear meniscus height (TMH) is easy after fluorescein installation, but the TMH after fluorescein instillation is higher than the TMH before fluorescein instillation. Therefore, we measured the time that the TMH after fluorescein instillation became the same with the TMH, we studied the difference in the TMH between normal eyes and dry eyes. The TMH 0.19 +/- 0.05 mm in the normal eyes and 0.10 +/- 0.04 mm in the dry eyes, and there was a significant difference between the 2 groups. The time that the TMH after fluorescein instillation became the same with the TMH before fluorescein instillation was 2.19 +/- 0.81 min. in the normal eyes and 2.29 +/- 0.73 min. in the dry eyes. Within 4 min. after fluorescein instillation, the TMH became the same with the TMH before fluorescein instillation in all cases and the height persisted until 7 min. after fluorescein instillation. Therefore, measuring the TMH will be easy if it is measured at 4-7 min. after fluorescein instillation.
Dry Eye Syndromes/*diagnosis ; Fluorescein ; Fluoresceins/*diagnostic use ; Humans ; Tears/*chemistry

Dry Eye is one of the most common eye disorders. Although this occurs in both men and women at any age, women in their menopausic years are often afflicted with this condition. This may be due to the loss of hormonal support after menopause. The purpose of this study is to determine the tear function of menopausic women and also to determine whether there is a correlation between advancing age and tear function. 62 menopausic women between the ages of 55-78 years underwent four different types of tear function tests (Schirmer Test, Tear Breakup Time, Vital Staining with Fluorescein Dye and the Ferning Test). These women were not under any type of medication. Results showed that in both the Schirmer Test and the Tear Breakup Time Test, there was a significant decrease in the tear function as age progressed. Although there were no statistically significant changes in the corneal uptake with increasing age, it was noted that the number of subjects with significant fluorescein dye uptake increased in the older age group. Likewise, there was no significant change in the Ferning test although a decreasing trend in the ferning pattern was observed as the age progressed. Based on the data collected, menopause and aging play a vital role in the development of dry eyes. Although the actual process remains uncertain, this can be attributed to the changes in the sex hormonal levels in menopausic women. (Author)
Human ; Female ; Aged ; Middle Aged ; HUMANS ; FEMALE ; POSTMENOPAUSE ; TEARS/CHEMISTRY ; LACRIMAL APPARATUS ; DRY EYE SYNDROME

OBJECTIVETo investigate the dynamic changes of matrix metalloproteinase-9 (MMP-9) level in tear fluid within 12 months after laser in situ keratomileusis (LASIK).

METHODSTwenty-two myopic patients undergoing uneventful LASIK were enrolled in this study. Tear fluid samples were collected from the patients for measurements of MMP-9 level using Western blotting preoperatively, at 7 and 14 days, and at 1, 2, 3, 6, and 12 months after the surgery.

RESULTSMMP-9 concentrations in the tear fluid of post-LASIK patients showed a time-dependent variation pattern. MMP-9 reached its peak level in the tear fluid at 14 days postoperatively, which was 2.70 times the preoperative level; it gradually decreased thereafter but was still 1.38 times the preoperative level at 12 months after the surgery.

CONCLUSIONSMMP-9 concentrations in the tear fluid of post-LASIK patients show a time-dependent variation pattern and remains higher than the preoperative level even at 12 months after the surgery, suggesting that corneal wound healing after LASIK lasts for more than 12 months.

Cornea ; Humans ; Keratomileusis, Laser In Situ ; Matrix Metalloproteinase 9 ; chemistry ; Myopia ; surgery ; Postoperative Period ; Prospective Studies ; Tears ; chemistry ; Wound Healing

OBJECTIVETo prepare eye drops of puerarin liposomes and investigate its lacrimal pharmacokinetics in rabbit tears.

METHODPuerarin liposomes were prepared by reverse phase evaporation technique. The liposomes and free puerarin were separated by SephadexG-50 chromatography and then encapsulation ratio of liposomes was determined in detail. Micromorphology of liposome particles was observed by electronic transmission microscope and the size distribution of the liposomes was analyzed by laser particle size analyzer. The concentration of puerarin in rabbit's tears was determined by HPLC after ocular administration of 50 microL puerarin liposomes while puerarin eye drops was chosen as control with the same puerarin concentration. The pharmacokinetic parameters were calculated by software program 3P97 according to one-compartment mode.

RESULTGlobal liposome nanoparticles with diameter of about 195.7 nm were prepared successfully. The encapsulation ratio of puerarin in the liposomes was 48.3%. The mean residence time (MRT) value and the area under concentration (AUC) of puerarin in liposome were 3.89 and 3.06 times more than those of puerarin eye drops.

CONCLUSIONLiposomes as a drug carrier can greatly increase the concentration of puerarin in tears, enhance the pre-ocular retention time than that of eye drops.

Animals ; Chromatography, High Pressure Liquid ; Isoflavones ; chemistry ; pharmacokinetics ; Liposomes ; chemistry ; pharmacokinetics ; ultrastructure ; Microscopy, Electron, Transmission ; Ophthalmic Solutions ; chemistry ; pharmacokinetics ; Rabbits ; Tears ; chemistry

The study is to design chitosan-coated pilocarpine nitrate submicro emulsion (CS-PN/SE) for the development of a novel mucoadhesive submicro emulsion, aiming to prolong the precorneal retention time and improve the ocular absorption. CS-PN/SE was fabricated in two steps: firstly, pilocarpine nitrate submicro emulsion (PN/SE) was prepared by high-speed shear with medium chain triglycerides (MCT) as oil phase and Tween 80 as the main emulsifier, and then incubated with chitosan (CS) acetic solution. The preparation process was optimized by central composite design-response surface methodology. Besides the particle size, zeta potential, entrapment efficiency and micromorphology were investigated, CS-PN/SE's precorneal residence properties and miotic effect were especially studied using New Zealand rabbits as the animal model. When CS-PN/SE was administered topically to rabbit eyes, the ocular clearance and the mean resident time (MRT) of pilocarpine nitrate were found to be dramatically improved (P < 0.05) compared with PN/SE and pilocarpine nitrate solution (PNs), since the K(CS-PN/SE) was declined to 0.006 4 +/- 0.000 3 min(-1) while MRT was prolonged up to 155.4 min. Pharmacodynamics results showed that the maximum miosis of CS-PN/SE was as high as 46.3%, while the miotic response lasted 480 min which is 255 min and 105 min longer than that of PNs and PN/SE, respectively. A larger area under the miotic percentage vs time curve (AUC) of CS-PN/SE was exhibited which is 1.6 folds and 1.2 folds as much as that of PNs and PN/SE, respectively (P < 0.05). Therefore, CS-PN/SE could enhance the duration of action and ocular bioavailability by improving the precorneal residence and ocular absorption significantly.
Absorption ; Animals ; Area Under Curve ; Biological Availability ; Chitosan ; chemistry ; Cornea ; metabolism ; Emulsions ; Microscopy, Electron, Transmission ; Miotics ; administration & dosage ; chemistry ; pharmacokinetics ; Ophthalmic Solutions ; Particle Size ; Pilocarpine ; administration & dosage ; chemistry ; pharmacokinetics ; Rabbits ; Random Allocation ; Solubility ; Tears ; metabolism

PURPOSE: To evaluate changes in clinical outcomes, inflammatory cytokine levels, and tear osmolarity in the tears of patients with moderate to severe dry eye syndrome before and after the application of topical 1% methylprednisolone. MATERIALS AND METHODS: Thirty-two patients with moderate to severe dry eye unresponsive to previous aqueous enhancement therapy were enrolled. Five patients were lost to follow up, and twenty-seven patients were eligible for analysis. Patients were instructed to apply topical 1% methylprednisolone four times per day, as well as to continue applying their current therapy of preservative-free 0.1% sodium hyaluronate four times per day. Corneal and conjunctival staining scores, tear film breakup time (TFBUT), Schirmer test, and tear osmolarity were assessed at baseline, 4 weeks, and 8 weeks. Tear samples were collected at every visit for cytokine analysis. RESULTS: Corneal and conjunctival staining scores and TFBUT showed significant improvement at 4 (p<0.001, <0.001, <0.001 respectively) and 8 (p<0.001, <0.001, <0.001 respectively) weeks. Tear osmolarity decreased significantly at 8 weeks (p=0.008). Interleukin (IL)-1beta, IL-8, and monocyte chemoattractant protein-1 were significantly decreased at 8 weeks compared with those at baseline (p=0.041, 0.001, 0.008 respectively). CONCLUSION: Short-term treatment with topical 1% methylprednisolone not only improved clinical outcomes, but also decreased tear osmolarity and cytokine levels. By measuring the changes in cytokine levels and tear osmolarity, we could objectively evaluate the anti-inflammatory effects of topical methylprednisolone applied in the treatment of patients with moderate to severe dry eye syndrome.
Adult ; Aged ; Cytokines/*metabolism ; Dry Eye Syndromes/*drug therapy/metabolism ; Female ; Humans ; Male ; Methylprednisolone/administration & dosage/*therapeutic use ; Middle Aged ; Osmolar Concentration ; Prospective Studies ; Tears/*chemistry/*metabolism

OBJECTIVETo investigate the efficiency of Spanishneedles Herb eye drops in treating perimenopausal xerophthalmia in rabbits.

METHODTotally 36 rabbits (36 right eyes) were ovariectomized, and 2 months later divided into three groups: the experimental group (group A, n = 12) given Spanishneedles Herb eye drops, the control group (group B, n = 12) given PBS and the model group (group C, n = 12) given no drug. The Schirmer I test (SIT), fluorescent (FL), total tear protein, diastase activity, lactoferrin and lysozyme contents and confocal scanning microscopy were performed at before the treatment and at 1 w, 2 w, 1 mo, 2 mo after the treatment.

RESULTBefore the treatment, There was no significant difference in SIT, FL, total tear protein, lysozyme, lactoferrin and amylase activity between two groups. Two months later after the treatment, both the group B and the group A showed differences degrees of changes in SIT, FL, total tear protein, lysozyme, lactoferrin and amylase activity compared with that before the treatment, with statistical differences (P < 0.05); At each time point, both groups revealed statistical differences in SIT, FL, total tear protein, lysozyme, lactoferrin and amylase activity (1 < 0.05). Two months later alter the treatment, densities of basal epithelial cells and inflammatory cells in the group A were (4 122 ±416) cells/mm2 and (339 ± 131) cells/mm2, while that in the group B were (3 343 ± 424) cells/mm2 and (49 ± 17) cells/mm2, with statistical differences between them (P < 0.05).

CONCLUSIONSpanishneedles Herb eye drops could effectively treat perimenopausal xerophthalmia in rabbit caused by sex hormones decline.

Animals ; Asteraceae ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Ophthalmic Solutions ; administration & dosage ; Perimenopause ; drug effects ; metabolism ; Rabbits ; Tears ; secretion ; Xerophthalmia ; drug therapy ; metabolism

PURPOSE: To investigate the properties of angiogenin (ANG) as a potential tool for the diagnosis and grading of dry eye syndrome (DES) by analyzing tear protein profiles. METHODS: Tear samples were collected with capillary tubes from 52 DES patients and 29 normal individuals as controls. Tear protein profiles were analyzed with an immunodot blot assay as a screening test. To confirm that the tear ANG levels were in inverse proportion to the disease severity grade, the ANG and lactoferrin (LF) tear contents of normal controls and DES patients were compared in an enzyme-linked immunosorbent assay. RESULTS: In the immunodot blot assay, the ANG area was lower in patients with grades 3 and 4 DES than in normal controls. The areas of basic fibroblast growth factor, transforming growth factor β2, and interleukin 10 were significantly greater than those of normal controls only in grade 4 DES patients, but these proteins were not linearly correlated with dry eye severity. Upon enzyme-linked immunosorbent assay analysis, the mean concentrations of ANG and LF decreased significantly as dry eye severity increased, except between grades 1 and 2. In addition, the ratios of ANG and LF to total tear proteins were correlated significantly with DES severity. CONCLUSIONS: ANG level was significantly lower in DES patients than in normal controls, and was significantly correlated with the worsening severity of DES, except between grades 1 and 2, as was LF. Therefore, ANG may be a useful measure of DES severity through proteomic analysis.
Adult ; Aged ; Angiogenesis Inducing Agents/pharmacology ; Dry Eye Syndromes/*diagnosis/metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Follow-Up Studies ; Humans ; Immunoblotting ; Male ; Middle Aged ; Proteomics/methods ; Ribonuclease, Pancreatic/*pharmacology ; Severity of Illness Index ; Tears/chemistry ; Young Adult

This study was performed to evaluate the changes of tear film and ocular surface caused by smoking. Symptom scoring, tear film break-up time (BUT), basal tear secretion test, corneal sensitivity test, keratoepitheliopathy scoring, and conjunctival impression cytology were performed in 29 smokers (58 eyes) and 26 non-smokers (52 eyes). Tear film BUT, basal tear secretion, corneal sensitivity, and squamous metaplasia were 7.71 +/- 2.66 sec, 6.29 +/- 2.85 mm, 53.69 +/- 5.69 mm, and 2.45 +/- 1.26 in smokers and 9.62 +/- 3.14 sec, 10.04 +/- 3.87 mm, 56.46 +/- 4.79 mm, and 1.12 +/- 0.83 in non-smokers, respectively (p< 0.05). Symptom score, keratoepitheliopathy score, and goblet cell density were not significantly different between the two groups. Tear film BUT was shorter, basal tear secretion and corneal sensitivity were lower, and squamous metaplasia was higher in heavy smokers than in light smokers. In conclusion, smoking deteriorates the tear film and ocular surface with decreased quantity and quality of tear film, decreased corneal sensitivity, and squamous metaplasia, and this deterioration is related to the amount of smoking.
Adult ; Aged ; Cell Count ; Conjunctiva/*metabolism/pathology ; Cornea/*metabolism/pathology ; Epithelial Cells/pathology ; Goblet Cells/metabolism/pathology ; Humans ; Lacrimal Apparatus/*metabolism/physiopathology ; Male ; Metaplasia ; Middle Aged ; Smoking/*metabolism/physiopathology ; Tears/chemistry/*secretion