OBJECTIVE: betaig-h3 is a 68kDa extracellular matrix protein which is overexpressed in synovial tissues of rheumatoid arthritis (RA). Previous results proved that betaig-h3 fragments are relevant to adhesion and migration of synovial fibroblast and angiogenesis through interaction with alphavbeta 3 integrin. We designed a recombinant betaig-h3 protein consisting of a fas-1 domain and RGD motif and evaluated the therapeutic efficacy in RA. METHODS: Inhibitory effect of adhesion and migration of NIH3T3 cell line was evaluated in 96 well microtiter and transwell plates coated with betaig-h3. Clinical arthritis index was evaluated after treating CIA mice with MFK12. Immunohistochemical staining in synovial tissues were performed. Expression of transcripts and proteins of inflammatory mediators were analyzed by semi-quantitative RT-PCR and immunoblotting. RESULTS: Recombinant protein consisted of 4th fas-1 domain truncated for H1 and H2 sequences and RGD peptide (MFK12), had M.W. of 10.4kDa. betaig-h3 mediated adhesion and migration of NIH3T3 cell line were significantly inhibited in a dose-dependent manner. Arthritis severity and incidence were efficiently reduced when CIA mice were treated with MFK12 at 30 mg/kg/day compared with the control. Immunohistochemical staining of joint tissues in MFK12 treated mice exhibited reduced angiogenesis. In treated mice, expression of transcripts regarding inflammatory mediators was markedly suppressed and immunoblotting of ICAM-1 and RANKL from whole extract of hind paws also showed a significant reduction. CONCLUSION: This study shows that MFK12 is effective in treating RA, although further study is warranted to improve the therapeutic efficacy.
Animals ; Arthritis ; Arthritis, Experimental ; Arthritis, Rheumatoid ; Cell Line ; Extracellular Matrix ; Extracellular Matrix Proteins ; Fibroblasts ; Immunoblotting ; Incidence ; Inflammation ; Intercellular Adhesion Molecule-1 ; Joints ; Mice ; Oligopeptides ; Proteins ; Transforming Growth Factor beta

BACKGROUND: Midazolam is often used as an anxiolytic premedication before surgery. But preoperatively administered midazolam may contribute to postopertive sedation and delayed recovery from general anesthesia. This study was undertaken to evaluate the effect of midazolam premedication on postoperative recovery and discharge-readiness after brief outpatient surgery. METHODS: Sixty healthy ASA physical status I women scheduled for outpatient diagnostic laparoscopic surgery were considered for the study. They were randomly allocated to one of two groups. Group one received normal saline (N/S) 5 ml intravenously (IV), while group two received IV midazolam 0.04 mg/kg. The study drug was prepared in 5 ml of saline and administered 10 minutes before the induction of general anesthesia. General anesthesia was induced with fentanyl, propofol and vecuronium and was maintained with N2O and enflurane. Postanesthetic recovery (PAR) scores were recorded after the arrival of the patients in the postanesthetic recovery room. Sedation was quantified before and after premedication and 60, 120 minutes after arriving in the postanesthetic recovery room, using the symbol-digit-modalities test (SDMT) and trail-making test (TMT). RESULTS: There were no significant differences between the two groups with respect to age, weight and anesthesia time. There were no significant differences in PAR scores or PAR-stay time between two groups. SDMT and TMT scores were significantly different 5 minutes after the study's drug administration, and 60 minutes after arrival in the postanesthetic recovery room between the two groups. The incidence of side effects was similar in both groups. CONCLUSIONS: Midazolam premedication proved effective in sedation and anxiolysis without prolonging postanesthetic recovery and discharge times for outpatient general anesthesia.
Ambulatory Surgical Procedures* ; Anesthesia ; Anesthesia, General ; Enflurane ; Female ; Fentanyl ; Humans ; Incidence ; Laparoscopy ; Midazolam* ; Outpatients* ; Premedication* ; Propofol ; Recovery Room* ; Vecuronium Bromide

OBJECTIVE: beta ig-h3 is an extracellular matrix protein, which is overexpressed in synovial tissues of rheumatoid arthritis (RA) similar to adhesive glycoproteins. We sought to evaluate the compensatory role of beta ig-h3 with adhesive glycoproteins in mediating the adhesion of fibroblast- like synoviocytes (FLS) and to confirm the inhibitory effect of YH18 peptide of the 2nd fas-1 domain in beta ig-h3-mediated adhesion. METHODS: The adhesion of FLS isolated from synovial tissues of RA, was evaluated in 96 well microtiter plate coated with matrix proteins. Inhibitory effect of YH18 peptides from the 2nd and 4th fas-1 domains was estimated in beta ig-h3-mediated adhesion of FLS. RESULTS: The adhesion of FLS on beta ig-h3 was weaker than that of fibronectin and vitronectin. The beta ig-h3-mediated adhesion was enhanced by the stimulation with phorbol myristate acetate (PMA), but not by cytokines and growth factors. Combination of fibronectin with beta ig-h3 synergistically enhanced the adhesion of FLS, in contrast to the additive effect of vitronectin combined with beta ig-h3. YH18 peptide of the 2nd fas-1 domain did not block the beta ig-h3-mediated adhesion of FLS. CONCLUSION: Our results reveal that beta ig-h3 may regulate the adhesion of FLS through the interaction with adhesive glycoproteins and confirm that the essential motifs mediating adhesion on beta ig-h3 are different according to the type of cells.