Hypoxia-ischemia leads to serious neuronal damage in some brain regions and is a strong risk factor for stroke. The aim of this study was to investigate the neuroprotective effect of tanshinone I (TsI) derived from Danshen (Radix Salvia miltiorrhiza root extract) against neuronal damage using a mouse model of cerebral hypoxia-ischemia. Brain infarction and neuronal damage were examined using 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin histochemistry, and Fluoro-Jade B histofluorescence. Pre-treatment with TsI (10 mg/kg) was associated with a significant reduction in infarct volume 1 day after hypoxia-ischemia was induced. In addition, TsI protected against hypoxia-ischemia-induced neuronal death in the ipsilateral region. Our present findings suggest that TsI has strong potential for neuroprotection against hypoxic-ischemic damage. These results may be used in research into new anti-stroke medications.
Animals ; Brain ; Brain Infarction ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; Eosine Yellowish-(YS) ; Fluoresceins ; Hematoxylin ; Hypoxia-Ischemia, Brain ; Mice ; Neurons ; Neuroprotective Agents ; Risk Factors ; Salvia miltiorrhiza ; Stroke ; Tetrazolium Salts

BACKGROUND: Though elastic fibers are as important as collagen fibers in interpretation of the histopathologic findings, it is impossible to observe them on the hematoxylin & eosin (H&E) stained specimen. OBJECTIVE: Characterizing eosin fluorescence emitted by elastic fibers in H&E stained specimens. METHODS: Normal skin tissue sections were stained in 4 different ways (unstained, hematoxylin only, eosin only, H&E) and observed under a fluorescence microscope using a FITC filter set. Fluorescent findings of 30 H&E-stained specimens showing abnormal dermal findings were compared with bright field findings of Miller's elastic stained specimen. RESULTS: Strong eosin fluorescence was related to the differential binding property of eosin with elastic fibers. Hematoxylin stain quenched excessive eosin fluorescence from other tissue components and contributed to better contrast. Fluorescence microscopy of H&E-stained sections was found to be especially useful in observing mature elastic fibers in the reticular dermis. In 74% of the specimens, eosin fluorescence findings of elastic fibers in reticular dermis matched well with that of specimens with elastic fiber special stain. CONCLUSION: Analysis of skin elastic fibers by fluorescence microscopy is a useful and complementary method to reveal hidden elastic fibers in H&E-stained specimens.
Collagen ; Dermis ; Elastic Tissue ; Enzyme Multiplied Immunoassay Technique ; Eosine Yellowish-(YS) ; Fluorescein-5-isothiocyanate ; Fluorescence ; Hematoxylin ; Microscopy, Fluorescence ; Skin

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

PURPOSE: The use of an autogenous fat graft has become a common procedure in plastic surgery. However, questions remain concerning on the viability of fat cells and preservation method of aspirated fat. The purpose of this study is to examine the viability of fat cells stored at -20degrees C in the freezer for 1 year after harvest from abdominal liposuction. METHODS: Eighteen adults(aged from 24 to 65 years, 16 female and 2 male) were selected for this study. Harvested aspirated fat tissues were obtained by suction-assisted lipectomy and frozen at -20degrees C commercial refrigerator for one year(average 12.5 months). The viability of fat cells in specimens were measured after thawing. The numbers of viable cells were measured on a fluorescence microscope after staining with fluorescein diacetate and propidium iodide. GPDH(Glycerol-3-phosphate dehydrogenase) activity was measured. Cell culture was done for 3 weeks. RESULTS: There were no viable cells under the fluorescence microscope, no detectable GPDH activity, and no cultured cells. CONCLUSION: These findings suggest that aspirated fat after frozen storage for one year at -20degrees C freezer is inadequate to reuse.
Adipocytes ; Cell Culture Techniques ; Cryopreservation ; Female ; Fluorescein ; Fluoresceins ; Fluorescence ; Humans ; Lipectomy ; Propidium ; Surgery, Plastic ; Transplants

PURPOSE: This study was performed to evaluate the parameters of tear function and ocular surface in patients with pinguecula and pterygium. METHODS: The corneal sensitivity test (CST), tear break-up time (BUT), basal tear secretion test, fluorescein staining, rose bengal staining, tear clearance test, and conjunctival impression cytology were evaluated in patients with unilateral pinguecula and pterygium. The results were also evaluated according to the severity of pterygium. RESULTS In patients with pinguecula, BUT (P=0.03) and goblet cell density (P<0.01) were decreased, while the rose bengal staining score (P=0.01) was increased significantly. In patients with pterygium, CST (P=0.01), BUT (P<0.01), and goblet cell density (P<0.01) decreased, and the fluorescein staining score (P<0.01), rose bengal staining score (P<0.01) and grade of conjunctival metaplasia (P<0.01) increased significantly. In comparison with mild pterygium, moderate pterygium demonstrated decreased CST (P=0.01) and BUT (P=0.01), and an increased rose bengal staining score (P=0.04). CONCLUSIONS: The tear films and ocular surface changes in patients with pinguecula and pterygium are worse than in control group. Also, there is close correlation between the severity of pterygium and dry eye condition.
Fluorescein ; Goblet Cells ; Humans ; Metaplasia ; Pinguecula* ; Pterygium* ; Rose Bengal ; Tears*

PURPOSE: This study was performed to evaluate the parameters of tear function and ocular surface in patients with pinguecula and pterygium. METHODS: The corneal sensitivity test (CST), tear break-up time (BUT), basal tear secretion test, fluorescein staining, rose bengal staining, tear clearance test, and conjunctival impression cytology were evaluated in patients with unilateral pinguecula and pterygium. The results were also evaluated according to the severity of pterygium. RESULTS In patients with pinguecula, BUT (P=0.03) and goblet cell density (P<0.01) were decreased, while the rose bengal staining score (P=0.01) was increased significantly. In patients with pterygium, CST (P=0.01), BUT (P<0.01), and goblet cell density (P<0.01) decreased, and the fluorescein staining score (P<0.01), rose bengal staining score (P<0.01) and grade of conjunctival metaplasia (P<0.01) increased significantly. In comparison with mild pterygium, moderate pterygium demonstrated decreased CST (P=0.01) and BUT (P=0.01), and an increased rose bengal staining score (P=0.04). CONCLUSIONS: The tear films and ocular surface changes in patients with pinguecula and pterygium are worse than in control group. Also, there is close correlation between the severity of pterygium and dry eye condition.
Fluorescein ; Goblet Cells ; Humans ; Metaplasia ; Pinguecula* ; Pterygium* ; Rose Bengal ; Tears*

OBJECTIVE: To evaluate the effect of intense pulsed light (IPL) in the treatment of chronic hordeolum. METHODS: Patients with chronic hordeolum who underwent IPL treatment were enrolled in this study. According to the severity of hordeolum, the patients were treated with IPL 3 to 5 times. Patients' satisfaction and visual analog scale scores for ocular discomfort symptoms before and after treatment were collected. The number, congestion, long diameter, short diameter and area of nodules were also recorded and measured. Finally, eyelid margin signs, meibum quality, meibomian gland expressibility, meibomian gland dropout, tear meniscus height, and corneal fluorescein staining were scored. RESULTS: 20 patients were enrolled in this study. The eyelid margins were congestive and swollen, with blunt rounding or irregularity. The meibum was cloudy or toothpaste-like. The meibomian gland expressibility, meibomian gland dropout and tear meniscus height were reduced. The cornea showed scattered fluorescein staining. After treatment, score of visual analog scale, congestion and size of nodules were significantly reduced. Eyelid margin signs, meibum quality, meibomian gland expressibility, tear meniscus height and corneal fluorescein staining scores were improved. Meibomian gland dropout had no significant change. No side effects occurred during treatment. CONCLUSIONS IPL is beneficial for the treatment of chronic hordeolum.
Humans ; Hordeolum ; Meibomian Glands ; Tears ; Fluoresceins

Cyclodialysis and ciliochoroidal detachment were performed in three eyes of three rabbits and in three eyes of another three rabbits, respectively. After aspiration of the aqueous humor, 0.1 ml of 10% sodium fluorescein was injected intracamerally, and the eyeball was enucleated between 30 minutes and one hour after injection and prepared for fluorescence microscopy. Sodium fluorescein concentrations in the supraciliary space were much greater in the group with cyclodialysis or ciliochoroidal detachment than in the normal control group. These results suggest that (1) in the eye with cyclodialysis, the aqueous humor may freely gain access to the supraciliary space through the cleft between the anterior chamber and the supraciliary space and then be removed rapidly and (2) in the eye with ciliochoroidal detachment, the aqueous humor may pass through the uveoscleral outflow pathway.
Animals ; Anterior Chamber/metabolism ; Aqueous Humor/*secretion ; Choroid/metabolism/*surgery ; Ciliary Body/metabolism/*surgery ; Fluorescein ; Fluoresceins/diagnostic use ; Microscopy, Fluorescence ; Rabbits

PURPOSE: To identify significant fluorescein angiographic (FA) characteristics associated with visual acuity (VA) in Behcet retinal vasculitis. MATERIALS AND METHODS: Retrospective review of 86 eyes of 48 patients (age: 35.6+/-10.2 years) with Behcet retinal vasculitis were performed. VA and FA findings as well as correlation between them were assessed. RESULTS: The mean initial VA of eyes with posterior pole-involved vasculitis (63 eyes; 73.3%) was significantly worse than that of those with peripheral vasculitis (23 eye; 26.7%) (logarithm of the minimum angle of resolution VA: 0.554+/-0.572 vs. 0.078+/-0.148; p<0.0001). Subgroup analysis revealed a more severe and diffuse pattern of vascular leakage in posterior pole-involved vasculitis compared to peripheral vasculitis (p<0.0001). Retinal vascular leakage (beta=0.345; p<0.0001), optic disc hyperfluorescence (beta=0.147; p=0.032), and macular leakage (beta=0.107; p=0.047) were significantly associated with worse initial VA. During the follow up (mean: 33.3+/-17.9 months), the change of leakage showed no significant correlation with change of VA in posterior pole-involved vasculitis (tau=0.199, p=0.092). CONCLUSION: Posterior pole involvement, the degree of retinal vascular leakage, optic disc hyperfluorescence, and macular leakage are significantly associated with VA in Behcet retinal vasculitis.
Adult ; Aged ; Behcet Syndrome/*diagnosis ; Female ; *Fluorescein Angiography ; Fluoresceins ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Regression Analysis ; Retinal Vasculitis/*diagnosis/physiopathology ; Retrospective Studies ; Visual Acuity/*physiology

PURPOSE: To evaluate the changes of tear film and ocular surface in keratoconus patients. METHODS: Twenty-eight eyes of 18 patients with keratoconus and 32 eyes of 16 normal subjects were included. We performed corneal sensitivity test, tear film break-up time (BUT), Schirmer test, fluorescein and rose bengal staining, and conjunctival impression cytology, and compared the results between the two groups. According to the central corneal power, we classified the patients into 3 groups and compared these parameters between groups. RESULTS: Corneal sensitivity was 5.56 +/- 0.34 mm and 5.88 +/- 0.25 mm (p<0.001), BUT was 9.64 +/- 2.51 sec and 10.69 +/- 2.89 sec (p=0.008), basal tear secretion was 9.00 +/- 3.06 mm and 10.88 +/- 3.95 mm (p=0.025), fluorescein staining score was 1.43 +/- 1.87 and 0.69 +/- 1.20 (p=0.105), and rose bengal staining score was 0.86 +/- 0.97 and 0.22 +/- 0.55 (p=0.003) in the patients and control subjects, respectively. The degree of conjunctival squamous metaplasia was 1.04 +/- 0.51 and 0.62 +/- 0.43 (p=0.039), and goblet cell density was 762.65 +/- 76.32 cells/mm2 and 880.21 +/- 82.06 cells/mm2 (p=0.041) in the patients and control subjects respectively. All parameters except basal tear secretion and fluorescein staining score were related to the severity of keratoconus. CONCLUSIONS: Tear film and ocular surface changes in patients with keratoconus are composed of decreased corneal sensitivity, decreased quantity and quality of tear film, and damages to corneal and conjunctival epitheliums, which are similar to the findings in keratoconjunctivitis sicca.
Epithelium ; Fluorescein ; Goblet Cells ; Humans ; Keratoconjunctivitis Sicca ; Keratoconus* ; Metaplasia ; Rose Bengal ; Tears*