Objective To compare the diagnostic value of fluorescent staining versus KOH wet-mount microscopy in detecting superficial fungal infection.Methods Totally,600 specimens from cases of clinically diagnosed superficial fungal infections and 102 from cases of clinically diagnosed Malassezia infection (including 54 cases of pityriasis versicolor and 48 cases of Malassezia folliculitis) were collected from the dermatology clinic of Tenth People's Hospital of Tongji University between July 2017 and February 2018.These specimens were subjected to fluorescent staining and KOH wet mount separately followed by direct microscopy,and the positive rate and average time for slide reading were compared between the two methods.Culture served as the gold standard method,and the missed diagnosis rate was compared between the two methods.Statistical analysis was carried out using chi-square test or Fisher's exact test for comparing enumeration data,and paired t test for comparing emeasurement data.Results Of the 600 specimens from clinically diagnosed superficial fungal infection cases,fungi were detected in 546 (91.00％) and 489 (81.50％) by fluorescent staining and KOH wet-mount microscopy respectively (x2 =22.83,P ＜ 0.05).Fluorescent staining showed significantly shorter average reading time (73.67 ± 13.56 s)compared with KOH wet-mount microscopy (87.12 ± 15.83 s,t =14.60,P ＜ 0.05).Among the 54 specimens from pityriasis versicolor cases,fluorescent staining and KOH wet-mount microscopy positive results in 51 (94.44％) and 50 (92.59％) specimens respectively (adjusted x2 =0,P ＞ 0.05),with the average reading time being 38.36 ± 8.79 s and 41.25 ± 15.67 s respectively (t =1.14,P ＞ 0.05).Of the 48 specimens from Malassezia infection cases,43 (89.58％) and 11 (22.92％) specimens were detected to be positive for fungi by fluorescent staining and KOH wet-mount microscopy respectively (x2 =43.34,P ＜ 0.05),and fluorescent staining showed shorter average reading time (42.14 ± 12.61 s) compared with KOH wet-mount microscopy (103.56 ± 9.48 s,t =17.83,P ＜ 0.05).Among the 600 specimens from superficial fungal infection cases,culture yielded fungi in 479.Moreover,476 specimens were found positive by fluorescent staining,and 3 were found negative (0.63％),while KOH wet-mount microscopy showed 465 positive results and 14 negative results (2.92％).There was a significant difference in the missed diagnosis rate between the two methods (x2 =7.25,P ＜ 0.05).Conclusion Compared with KOH wet-mount microscopy,fluorescent staining can increase the detection rate,reduce missed diagnosis rate and shorten reading time.

OBJECTIVETo test whether splicing overlapping extension(SOE) method can be a tool for obtaining rare fusion gene's transcripts and to study the tumorigenic capacity of a novel fusion gene AML1-MTG16.

METHODSSOE method was used to obtain AML1- MTG16 fusion gene's transcripts. MTG16, AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 segment were inserted into pEGFP- C1,pDsRed-N1 vector respectively,then transfected NIH3T3 cell line by lipofection. Forty-eight hours later, the transfected cells were examined by laser-scanning confocal microscopy. Stable transfected cells were obtained by G418 500ug/ul selection for one month. Growth curve, soft agar colonies formation tumorigenesis in nude mice were done to compare the difference between stable transfected cells.

RESULTSRecombined AML1-MTG16 by SOE contained its CDS. NIH3T3 expressing AML1-MTG16 had a faster proliferation in medium, colony growth in soft agar. AML1-MTG16 expression cells also induced tumors formation following injection into nude mouse. MTG16,AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 were colocalized in the nucleus of cotransfected NIH3T3 cells under the examination of laser-scanning confocal microscope.

CONCLUSIONSOE is an effective method to get rare fusion gene's transcripts. AML1-MTG16 plays an important role in leukemogenesis. MTG16 may also have a carcinogenic property within the AML1-MTG16 fusion gene. Carcinogenic property of AML1-MTG16 is restricted to its localization in the nuclear matrix. N terminal of MTG16 may play an important part in the carcinogenic activity of AML1-MTG16.

3T3 Cells ; transplantation ; Animals ; Cell Division ; genetics ; Cell Transformation, Neoplastic ; genetics ; Cell Transplantation ; Core Binding Factor Alpha 2 Subunit ; Green Fluorescent Proteins ; Luminescent Proteins ; genetics ; metabolism ; Mice ; Mice, Nude ; Microscopy, Confocal ; Neoplasms, Experimental ; genetics ; pathology ; Oncogene Proteins, Fusion ; genetics ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Transfection