Resveratrol has neuroprotective effects and plays an anti-inflammatory role through Dectin-1/p38 pathway in Aspergillus fumigatus keratitis.

PURPOSE: To determine the antifungal, anti-inflammatory and neuroprotective effects of resveratrol (RES) in Aspergillus fumigatus (A. fumigatus) keratitis. METHODS: Cytotoxicity assay and Draize eye assay were performed to assess the toxicity of RES. The antifungal effect of RES was assessed by minimal inhibitory concentration, scanning or transmission electron microscopy, propidium iodide uptake assay, and Calcofluor white staining. Phosphorylation of p38 MAPK, mRNA and protein levels of Dectin-1 and related inflammatory factors were measured by qRT-PCR, ELISA and Western blot in vitro and in vivo. Clinical score, HE staining, plate count, and myeloperoxidase test were used to observe the progress of fungal keratitis. IF staining, qRT-PCR, and the Von Frey test were selected to assess the neuroprotective effects of RES. RESULTS: RES suppressed A. fumigatus hyphae growth and altered hyphae morphology in vitro. RES decreased the expression of Dectin-1, IL-1ß and TNF-α, as well as p38 MAPK phosphorylation expression, and also decreased clinical scores, reduced inflammatory cell infiltration and neutrophil activity, and decreased fungal load. RES also protected corneal basal nerve fibers, down-regulated mechanosensitivity thresholds, and increased the mRNA levels of CGRP and TRPV-1.. CONCLUSION: These evidences revealed that RES could exert antifungal effects on A. fumigatus and ameliorate FK through suppressing the Dectin-1/p38 MAPK pathway to down-regulate IL-1ß, IL-6, etc. expression and play protective effect on corneal nerves.

Benzyl isothiocyanate improves the prognosis of Aspergillus fumigatus keratitis by reducing fungal load and inhibiting Mincle signal pathway.

Aspergillus fumigatus keratitis is a potential blinding disease associated with A. fumigatus invasion and excessive inflammatory response. Benzyl isothiocyanate (BITC) is a secondary metabolite with broad antibacterial and anti-inflammatory activity extracted from cruciferous species. However, the role of BITC in A. fumigatus keratitis has not been discovered yet. This study aims to explore the antifungal and anti-inflammatory effects and mechanisms of BITC in A. fumigatus keratitis. Our results provided evidences that BITC exerted antifungal effects against A. fumigatus by damaging cell membranes, mitochondria, adhesion, and biofilms in a concentration-dependent manner. In vivo, fungal load and inflammatory response including inflammatory cell infiltration and pro-inflammatory cytokine expression were reduced in BITC-treated A. fumigatus keratitis. Additionally, BITC significantly decreased Mincle, IL-1ß, TNF-α, and IL-6 expression in RAW264.7 cells that stimulated by A. fumigatus or Mincle ligand trehalose-6,6-dibehenate. In summary, BITC possessed fungicidal activities and could improve the prognosis of A. fumigatus keratitis by reducing fungal load and inhibiting the inflammatory response mediated by Mincle.

Pentoxifylline treats Aspergillus fumigatus keratitis by reducing fungal burden and suppressing corneal inflammation.

Fungal keratitis (FK) is a blinding ocular disease, which mainly results from fungal damage and excessive inflammation. Pentoxifylline, a kind of methylxanthine, has been discovered to have anti-inflammatory properties in various infectious diseases, hinting a potential therapeutic effect on treating corneal fungal infection. Whereas, the therapeutic impact of pentoxifylline on fungal keratitis is still uncertain. This study investigated the antifungal capability against Aspergillus fumigatus and the anti-inflammatory role of pentoxifylline by activating nuclear factor, erythroid 2 like 2 (Nrf2)/heme oxygenase1 (HO1) pathway in the process of FK. In our research, we demonstrated that pentoxifylline could effectively inhibit fungal growth and inflammatory reaction. Pentoxifylline reduced the production of pro-inflammatory factors by stimulating the Nrf2/HO1 pathway. Although there was no statistical difference between the curative efficacy of pentoxifylline and natamycin application to FK, pentoxifylline could promote corneal epithelial repair and was less toxicity to the ocular surface than natamycin. In conclusion, pentoxifylline performs antifungal and anti-inflammatory effects by lessening the fungus burden and activating the Nrf2/HO1 pathway, hinting that it has the potential to be a new therapeutic medication for Aspergillus fumigatus keratitis.

The therapeutic role and mechanism of 4-Methoxycinnamic acid in fungal keratitis.

Fungal keratitis is an infectious vision-threatening disease that has a poor prognosis, and the clinical therapeutic drugs have multiple limitations, such as epithelial toxicity and low bioavailability. Therefore, new antifungal treatment strategies must be developed. 4-Methoxycinnamic acid (MCA) is a widely occurring natural phenolic acid that has been proven to have multiple effects, such as antibacterial, antifungal, anti-inflammatory, neuroprotective, and inhibiting cancer. In this research, we explored the effects and underlying mechanisms of MCA on A. fumigatus keratitis and the antifungal effects of the combination of MCA and natamycin (NATA) on A. fumigatus. We found that MCA exerts antifungal effects by inhibiting the synthesis of the fungal cell wall, changing the permeability of fungal cell membranes. Moreover, the MCA-NATA combination exhibited synergy for A. fumigatus. In addition, MCA exerted an anti-inflammatory effect by downregulating the inflammatory factors (IL-1ß, TNF-α, IL-6, and iNOS) in C57BL/6 mice and RAW264.7 cells. The anti-inflammatory mechanism of MCA was associated with the Mincle signal pathway. In summary, MCA acts as a potential therapeutic drug for fungal keratitis and a potential antifungal sensitizer for natamycin. MCA inhibits fungal cell wall synthesis, destroys the permeability of fungal cell membranes, and mediates the anti-inflammatory, immune response of the host.

[Applied anatomy of the path of retroperitoneal laparoscopy surgery].

OBJECTIVE: To discover the anatomical basis of retroperitoneal laparoscopic surgery. METHODS: Twenty Chinese adult cadavers including 12 and 8 woman were anatomized. The relationship between nerves, vessels of the lateral abdominal wall, retroperitoneal structures and the path of retroperitoneal laparoscopic surgery were measured. RESULTS: In male cadavers, the distance between the iliac crest and lumbar trigonum was 4.02 +/- 1.26 cm on the left, and 3.83 +/- 0.90 cm an the right. The distance between the iliac crest and ilio-hypogastric nerve was 0.82 +/- 0.13 cm on the left and 0.84 +/- 0.08 cm on the right. The distance between the subcostal nerve and XII rib in the posterior axillary line and midaxilla line was 0.79 +/- 0.26 cm, 1.65 +/- 1.12 cm on the left and 0.78 +/- 0.30 cm, 1.59 +/- 1.07 cm on the right respectively. The distance between the ureter of inferior pole of kidney and extra-border of psoas was 2.24 +/- 0.67 cm on the left and 2.19 +/- 0.73 cm on the right. The distance between crossing of the ureter and iliac vessels and extra-border of psoas was 3.91 +/- 0.59 cm on the left and 3.76 +/- 0.53 cm on the right. In female cadavers, the distance between the iliac crest and trigonum lumbale was 3.90 +/- 0.37 cm on the left and 3.87 +/- 0.37 cm on the right. The distance between the iliac crest and ilio-hypogastric nerve was 0.94 +/- 0.06 cm on the left and 0.93 +/- 0.10 cm on the right. The distance between the subcostal nerve and XII rib in the posterior axillary line and midaxilla line was 0.61 +/- 0.14 cm and 1.37 +/- 0.89 cm on the left and 0.64 +/- 0.13 cm, 1.32 +/- 0.95 cm on the right respectively. The distance between the ureter of inferior pole of kidney and extra-border of psoas was 2.00 +/- 0.23 cm on the left and 1.91 +/- 0.13 cm on the right. The distance between crossing of the ureter and iliac vessels and extra-border of psoas was 3.43 +/- 0.31 cm on the left and 3.85 +/- 0.43 cm on the right. CONCLUSIONS: The top of the lumbar trigonum which is 2 cm above the iliac crest selected as the first point and 2 cm inferior to the XII rib in the posterior axillary line selected as the second point is the anatomical basis for retroperitoneal laparoscopic surgery. It can reduce the opportunity to injury nerves, vessels and retroperitoneal structures while making retroperitoneal laparoscopic surgery easy.