Clinical Trial: The Pathogenesis of Superior Limbic Keratoconjunctivitis
Study Status: Completed
Recruit Status: Unknown status
Study Type: Observational
Official Title:
Brief Summary: Superior limbic keratoconjunctivitis (SLK) was first described in detail as a clinical entity by Frederick Theodore in 1963. The clinical picture of SLK is well documented, but the etiology is still unknown. This project will be conducted into through two parts: one is to investigate the presentation of chemokine receptors on mast cell and matrix metalloproteinases on fibroblasts by immunohistochemistry method from the pathological specimens of SLK patients who received conjunctiva resection as the treatment. The other part is to investigate the mRNA level of those chemokine receptors via reverse transcription - polymerase chain reaction from the conjunctiva collecting form SLK patients.
Detailed Summary:
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Specimens collection:
We will collect the specimens in two different parts. One part is to collect all the paraffinized block of the SLK patients to get fifteen copies of 5-μm section of each paraffinized block from the Department of Pathology. All the slides will be deparaffinized and rehydration for further immunohistochemistry (IHC) stain . Primary antibodies for IHC stain include chemokine receptors (CXCR1, CXCR2, CXCR3, CXCR4, CCR1. CCR3, CCR4, CCR5, CD30L) and matrix metalloproteinases (MMP-1, -2, -3, -9).
Another part of this study is to collect the conjunctiva of SLK patients and normal control patients. This part of the clinical study is sent for IRB approval concomitantly. We will collect the conjunctiva of SLK patients when they receive conjunctival resection as the treatment in the following one year. And the normal control conjunctiva will be obtained while the patients come to our hospital for cataract surgery or retinal surgery with redundant conjunctiva noted after peritomy. The fresh conjunctiva will be stored in -80℃ before RNA isolation. All the tissue will be processed into RNA and reverse transcripted into cDNA. Polymerase chain reaction will be done with the primers, including CXCR1, CXCR2, CXCR3, CXCR4, CCR1, CCR3, CCR4, CCR5, and CD30L.
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Preparation of RNA and cDNA:
Total RNA will be extracted from the conjunctiva of SLK and control groups with Trizol reagent (Life, Gaithersburg, MD, USA). One microgram of total RNA from each sample will be annealed for 5 min at 70℃ with 500 ng oligo(dT) (Fermentas, Hanover, MD, USA) and reverse transcribed to cDNA by 200u RevertAid™
Sponsor: National Taiwan University Hospital
Current Primary Outcome:
Original Primary Outcome:
Current Secondary Outcome:
Original Secondary Outcome:
Information By: National Taiwan University Hospital
Dates:
Date Received: September 12, 2005
Date Started: July 2005
Date Completion: June 2006
Last Updated: November 25, 2005
Last Verified: June 2005
