Clinical evaluation of rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy / 中华实验眼科杂志

Background The rapid diagnosis can win more treating opportunities for patients with fungal keratitis.Even though the fungal culture is the gold standard for the diagnosis of fungal keratitis,it is difficult in early diagnosis due to the long duration of cultivation and false-negative rate.Objective This trial was to explore the clinical value in the rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy.Methods Corneal scraping and laser scanning confocal microscopy were separately performed in 167 eyes of 167 patients with fungal keratitis.All the eyes were examined by the slit lamp,followed by laser scanning confocal microscope,and then the 10％ KOH corneal smear was examined under the optical microscope.Results The positive rate of diagnosis was 75％ (125/167) by corneal scraping,and that by laser scanning confocal microscopy was 91％ (152/167).The positive rate of examining outcome was significantly higher in laser scanning confocal microscopy than that of corneal scraping (x2 =14.88,P =0.00).The positive results were 114 cases and negative results were 4 cases by two methods,with the concordance rate 70.7％ (118/167).The hyphae or spore were seen in 32 cases by laser scanning confocal microscopy in 42 negative cases by corneal scraping,and in 15 negative cases by confocal laser scanning microscopy,11 positive outcomes were offered by corneal scraping.Conclusions The combined application of corneal scraping with confocal laser scanning microscopy can improve and speed up the diagnosis positive rate of fungal keratitis.

Cloning, expression and functional analyses of human platelet-derived growth factor-B chain peptide for wound repair of cat corneal endothelial cells / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the biological function of platelet-derived growth factor B (PDGF-B) on the survival and proliferation of cat corneal endothelial cells so as to provide bases for further studies of its role in wound repair and its clinical application.</p><p><b>METHODS</b>Total RNA was extracted from the placenta tissues of healthy pregnant women undergoing hysterotokotomy and PDGF cDNA was obtained with reverse transcription-polymerase chain reaction (RT-PCR). The prokaryotic expression vector pET-PDGF-B was constructed and expressed the recombinant PDGF-B in Escherichia coli (E. coli) BL21 (DE3). After purification and refolding on Ni2+-chelation affinity chromatography (NTA) column, it was used to culture cat corneal endothelial cells. Cell proliferation was tested by modified tertrazolium salt (MTT) and flow cytometer. And the morphologic change and the ultrastructure were observed under an inverted phase contrast microscope, a scanning electron microscope and a transmission electon microscope, respectively.</p><p><b>RESULTS</b>PDGF-B chain peptide (PDGF-BB) gene was successfully inserted into the prokaryotic expression vector, pET-28a (+). The purified recombined protein pET-PDGF-B showed a single band on sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) with the molecular weight of about 27 u, which was in agreement with the deduced value. MTT and flow cytometry showed that PDGF-BB promoted the survival and proliferation of cat corneal endothelial cells.</p><p><b>CONCLUSIONS</b>The construction of recombinant prokaryotic expression vector pET-PDGF-B and the preparation of PDGF-BB protein provide a foundation for further study of the function of PDGF-BB and producing biological PDGF-BB protein. The expressed PDGF-BB promotes the proliferation of cultured cat corneal endothelial cells.</p>

Animal study on expression of laminin and fibronectin in cornea during wound healing following alkali burn / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To observe the expression of laminin and fibronectin in alkali-burned corneas in rats.</p><p><b>METHODS</b>A total of 18 normal Wistar rats were randomly divided into 6 groups (n = 3 in each group). For each rat, one eye was injured by alkali burn, the other one was taken as the normal control. Then all the corneas were surgically removed and the expression of laminin and fibronectin was observed with immunohistochemistry respectively at 7 hours, 1 day, 3 days, 7 days, 14 days and 28 days after alkali burn.</p><p><b>RESULTS</b>Compared with that of the normal controls, the expression of laminin and fibronectin of the burned eyes was dramatically higher at 7 hours, reached peak at 14 days and decreased to the normal level at 28 days after alkali burn.</p><p><b>CONCLUSIONS</b>In the process of wound healing after alkali burn, the expression of laminin and fibronectin increases dramatically, which suggests that laminin and fibronectin may participate in the process of corneal wound healing.</p>