Fusarium solani Activates Dectin-1 in Experimentally Induced Keratomycosis.

In this study, the response of Dectin-1 on macrophages to Fusarium solani (F. solani) and the expression patterns of Dectin-1 in experimentally F. solani-induced keratomycosis were investigated. Peritoneal macrophages isolated after intraperitoneal injection of sodium thioglycollate were co-cultured with laminarin and spores of F. solani for 12 h. The expression levels of Dectin-1 and CARD9 were detected by immunofluorescence and real-time quantitative polymerase chain reaction. A mouse model of fungal keratitis was established by substromal inoculation with spores of F. solani. Corneal lesions and inflammatory responses were observed by slit-lamp and histopathology at 1, 2, 3, 5, and 7 day(s) after infection. Dectin-1 expression was significantly upregulated on macrophages stimulated by spores of F. solani. Dectin-1 was not detected in normal corneas of C57BL/6 mice, but detected in infected corneas from the first day after inoculation, with high mRNA levels observed on days 2 and 3. CARD9, a key transducer of Dectin-1 signaling, was also upregulated in infected corneas. In conclusion, Dectin-1 is an important recognition receptor in F. solani-induced keratitis, but the molecular mechanisms warrant further investigation.

[Expression and its promoter methylation of chemokine CXC ligand 14 in peripheral blood mononuclear cells from patients with lupus].

OBJECTIVE: To analyze the expression and its promoter methylation of chemokine CXC ligand 14 (CXCL14) in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE). METHODS: The RNAs of PBMCs from 28 SLE patients and 20 healthy controls were isolated and reversely transcribed into cDNAs. Using GAPDH as the internal reference, the levels of CXCL14 ex-pression were detected by real-time polymerase chain reaction (PCR). The correlation between CXCL14 expression and the clinic pathological fe atures of SLE were further analyzed. DNA methylation was analyzed by bisulfite sequencing PCR (BSP). RESULTS: Our data indicated that the level of CXCL14 in the PBMC of SLE patients was statistically lower than that in healthy controls (P < 0.05). Further analysis showed that CXCL14 expression was negatively correlated with anti-Sj gren syndrome B antibody(anti-SSB antibody, P < 0.01) and albuminuria(P < 0.05). However, CXCL14 expression was not significantly correlated with the indexes of SLE activity, renal damage, the level of anti-ds-DNA antibodies, complement C3 and C-reactive protein. In addition, we further demonstrated that the CXCL14 promoter hypermethylation expres-sion was significant higher than healthy controls. CONCLUSIONS: Down-regulated of CXCL14 expression in PBMC maybe involved in the occur-rence or development of SLE disease. The loss of CXCL14 expression was regulated by promoter hypermethylation.

Amniotic membrane covering promotes healing of cornea epithelium and improves visual acuity after debridement for fungal keratitis.

AIM: To investigate the effect of amniotic membrane covering (AMC) on the healing of cornea epithelium and visual acuity for fungal keratitis after debridement. METHODS: Twenty fungal keratitis patients were divided into two groups randomly, the AMC group and the control group, ten patients each group. Both debridement of the infected cornea tissue and standard anti-fungus drugs treatments were given to every patients, monolayer amniotic membrane were sutured to the surface of the entire cornea and bulbar conjunctiva with 10-0 nylon suture for patients in the AMC group. The diameter of the ulcer was determined with slit lamp microscope and the depth of the infiltration was determined with anterior segment optical coherence tomography. Uncorrected visual acuity (UCVA) was tested before surgery and three month after healing of the epithelial layer. The healing time of the cornea epithelium, visual acuity (VA) was compared between the two groups using t-test. RESULTS: There was no statistical difference of the diameter of the ulcer, depth of the infiltration, height of the hypopyon and VA between the two groups before surgery (P>0.05). The average healing time of the AMC group was 6.89±2.98d, which was statistically shorter than that of the control group (10.23±2.78d) (P<0.05). The average UCVA of the AMC group was 0.138±0.083, which was statistically better than that of the control group (0.053±0.068) (P<0.05). CONCLUSION: AMC surgery could promote healing of cornea epithelium after debridement for fungal keratitis and lead to better VA outcome.

Promotion of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on titanium.

AIM: To investigate the influence of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on titanium (Ti) surface. METHODS: The chimeric peptide RKLPDAPRGDN (minTBP-1-PRGDN) was synthesized by connecting RKLPDA (minTBP-1) to the N-terminal of PRGDN, the influence of minTBP-1-PRGDN on the attachment, proliferation and collagen I synthesis of human keratocyte on Ti surface were tested using PRGDN and minTBP-1as controls. The keratocytes attached to the surface of Ti were either stained with FITC-labeled phalloidin and viewed with fluorescence microscope or quantified with alamar Blue method. The proliferation of keratocytes on Ti were quantified with 3-(4,5-dim- ethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide up-taking methods. The secretion of type I collagen were determined using an ELISA kit. RESULTS: The results showed that minTBP-1-PRGDN at a concentration of 100ng/mL was the most potent peptide to enhance the attachment of human keratocytes to the surface of Ti (1.40±0.03 folds, P=0.003), to promote the proliferation (1.26±0.05 folds, P=0.014) and the synthesis of type I collagen (1.530±0.128, P=0.008). MinTBP-1 at the same concentration could only promote the attachment (1.13±0.04 folds, P=0.020) and proliferation(1.15±0.06 folds, P=0.021), while PRGDN had no significant influence (P>0.05). CONCLUSION: Our data shows that the novel chimeric peptide minTBP-1-PRGDN could promote the attachment, proliferation and type I collagen synthesis of human keratocytes on the surface of Ti.

Hypopyon in patients with fungal keratitis.

BACKGROUND: Hypopyon is common in eyes with fungal keratitis. The evaluation of the clinical features, culture results and the risk factors for hypopyon and of the possible correlation between hypopyon and the treatment outcome could be helpful for making treatment decisions. METHODS: The medical records of 1066 inpatients (1069 eyes) with fungal keratitis seen at the Shandong Eye Institute from January 2000 to December 2009 were reviewed retrospectively for demographic features, risk factors, clinical characteristics, laboratory findings and treatment outcomes. The incidence of hypopyon, the fungal culture positivity for hypopyon, risk factors for hypopyon and the effect of hypopyon on the treatment and prognosis were determined. RESULTS: We identified 1069 eyes with fungal keratitis. Of the 850 fungal culture-positive eyes, the Fusarium species was the most frequent (73.6%), followed by Alternaria (10.0%) and Aspergillus (9.0%). Upon admission, 562 (52.6%) eyes with hypopyon were identified. The hypopyon of 66 eyes was evaluated via fungal culturing, and 31 eyes (47.0%) were positive. A total of 194 eyes had ocular hypertension, and 172 (88.7%) of these eyes had hypopyon (P < 0.001). Risk factors for incident hypopyon included long duration of symptoms (P < 0.001), large lesion size (P < 0.001) and infection caused by the Fusarium and Aspergillus species (P < 0.001). The positivity of fungal culture for hypopyon was associated with duration of symptoms and lesion size. Surgical intervention was more common in cases with hypopyon (P < 0.001). Hypopyon was a risk factor for the recurrence of fungal keratitis after corneal transplantation (P = 0.002). CONCLUSIONS: Hypopyon is common in patients with severe fungal keratitis and can cause ocular hypertension. About half of the hypopyon cases were positive based on fungal culture. Long duration of symptoms, large lesion size and infection with the Fusarium and Aspergillus species were risk factors for hypopyon. The presence of hypopyon increases the likelihood of surgical intervention.

Effects of endothelin-1 on the cytoskeleton protein F-actin of human trabecular meshwork cells in vitro.

AIM: To observe the effect of endothelin-1 (ET-1) on the cytoskeleton protein F-actin of cultured human trabecular meshwork (HTM) cells. METHODS: CULTURED HTM CELLS WERE RANDOMLY DIVIDED INTO FOUR GROUPS: control group, low-dose ET-1 (10(-9) mol/L) treatment group, middle-dose ET-1 (10(-8) mol/L) treatment group, and high-dose ET-1(10(-7) mol/L) treatment group. After treated with ET-1, the expression of cytoskeleton protein F-actin in trabecular meshwork was analyzed with Western-blot and the distribution of F-actin was detected with FITC-Phalloidin probe. RESULTS: ET-1 dose-dependently and significantly increased F-actin in trabecular meshwork cells (P<0.05). The F-actin stress fiber and periphery actin fiber highly increased and manifested mild reorganization after treated with ET-1; and there were much more cell-to-cell and cell-to-extracellular matrix attachments formation in ET-1 treated HTM cells than that in the untreated HTM cells. CONCLUSION: ET-1 promotes the expression of cytoske- leton protein F-actin and induced the trabecular meshwork actin cytoskeleton reorganization.