Facial cutaneous phaeohyphomycosis caused by Microsphaeropsis arundinis: a case report / 中华皮肤科杂志

A 55-year-old male patient presented with plaques on the face for more than 20 years,and no immunodeficiency diseases were diagnosed.Skin examination showed large areas of pink plaques on the nose,bilateral cheeks and upper oral lips with slight desquamation,verrucous hyperplasia on the dorsal area of the nose,and a bean-sized verrucous protuberance on the tip of the nose.Histopathological examination of the skin lesions revealed pseudoepitheliomatous hyperplasia in the epidermis and hyphae-like structures in the stratum corneum.Moreover,there was diffuse infiltration of inflammatory cells in the dermis,which mainly included neutrophils,lymphocytes,histiocytes and multinucleated giant cells.Periodic acid-Schiff (PAS)-positive spore-like structures were observed in the multinucleated giant cells.Culture of the lesional tissues on Sabouraud dextrose agar (SDA) medium showed grey-brown villous colonies.Microculture on the potato dextrose agar (PDA) medium yielded dark septate hyphae and pycnidia filled with a large number of spores.Microsphaeropsis arundinis was identified by fungal molecular biological techniques.The patient was diagnosed with cutaneous phaeohyphomycosis caused by Microsphaeropsis arundinis.The patient was treated with CO2 laser for the removal of verrucous protuberance on the tip of the nose,and oral itraconazole capsules at a dose of 200 mg twice a day.After 3-month treatment,the skin lesions subsided and the drug was withdrew.During 6-month follow-up,no relapse occurred.

Dynamic distribution of calmodulin labeled with red fluorescent protein in the polar growth of Aspergillus fumigatus / 中华皮肤科杂志

Objective To construct a native promoter-regulated Aspergillus fumigatus strain containing red fluorescent protein-labeled calmodulin (CaM-RFP),and to observe the dynamic distribution of calmodulin during the growth of Aspergillus fumigatus.Methods Bilateral flanking sequences of Aspergillus fumigatus calmodulin gene were designed,and plasmids containing the two flanking sequences and mRFP-Aspergillus fumigatus pyrG gene (mRFP-AfpyrG) were amplified separately.The final linear PCR product for transformation was generated from the above three PCR products by fusion PCR.Then,the above linear fragment was transferred into the Aspergillus fumigatus strain by protoplast transformation,so as to construct the CaM-RFP Aspergillus fumigatus strain.The monoclonal colony was picked from the screening medium and subjected to culture.Then,the stablest fluorescent monoxenic strain of Aspergillus fumigatus was selected,and the transformant was verified by PCR.The recombinant strain and wild-type stain were cultured on solid nutrient media separately,and the morphology of these strains was observed by fluorescence microscopy at different time points.Additionally,the above 2 strains were cultured in liquid media separately,and XTT assay was performed to evaluate the growth activity of strains.Microscopy was also conducted to dynamically observe the CaM-RFP Aspergillusfumigatus strain,and analyze the spatial and temporal distribution of calmodulin during the growth and development of Aspergillus fumigatus.Results The fluorescent phenotype and PCR identification results both indicated the successful construction of the CaM-RFP Aspergillus fumigatus strain.The growth activity at 24 hours did not differ between the recombinant strain and wild-type stain (A490:0.689 ± 0.081 vs.0.678 ± 0.054,t =1.32,P ＞0.05),so did the morphology.During the polarized growth of Aspergillus fumigatus,calmodulin was always at the top of the hyphae,germination site of the hyphal branch and the top of new branches.Conclusion Calmodulin may be involved in the regulation of spore germination and polar hyphal growth of Aspergillus fumigatus.

An albino strain of Aspergillus fumigatus causes pulmonary aspergilloma in an individual with normal immune ;system / 中华皮肤科杂志

A 76?year?old female patient complained of right chest pain for three months. CT scan showed a clump?like high?density shadow measuring 4.8 cm × 3.0 cm in size in the dorsal portion of the right lower lobe of the lung. Aspiration biopsy was performed, and biopsy samples were subjected to fungal culture and histopathological examination. Histopathological examination showed chronic granulomatous inflammation with hyaline septate hyphae. After 4?day culture, white villous dense colonies were formed on the Sabouraud′s agar medium. The center of the colonies was slightly elevated with wrinkles or radiating striae on the surface, and the bottom of the colonies was faint yellow in color. Microculture yielded abundant septate branched hyphae, and very few colorless hyaline quasi?circular spores. DNA sequencing of rDNA internal transcribed spacer (ITS) regions and β?tubulin genes was performed to identify the isolate, and antifungal susceptibility testing was carried out in vitro. The MEGA7.0 software was used to build phylogenetic trees of Aspergillus fumigatus complex and its closely related species. The isolate was identified as Aspergillus fumigatus by molecular biologic sequencing. The patient was diagnosed with pulmonary aspergilloma. After administration of itraconazole oral solution and vorionazole tablets, the condition got better obviously.

A case of cutaneous Arthrinium phaeospermum infection after lower limb amputation / 中华皮肤科杂志

A 59?year?old female patient, who received bilateral lower limb amputation 39 years ago, presented with eczematoid changes in both lower limbs for over 20 years, and with chronic granuloma?like lesions complicated by verrucous hyperplasia for more than 10 years. There were large areas of infiltrative and proliferative lesions with exudation and peripheral erythema at the amputation sites in both knee joints. The lesions were hard with tenderness on palpation. Microscopic examination of lesional scales with 10%KOH showed negative results for fungi. However, three times of culture on the Sabouraud dextrose agar(SDA)medium all grew the same kind of fungus, and the front side and reverse side of its filamentous colony were white and orange yellow respectively. Microculture showed that linear hyaline conidiophores came out from lageniform mother cells with conidia ascending alongside. The conidia looked like dark brown eye lens, with an equatorial germ slit. Based on these findings, this fungus was identified as Arthrinium phaeospermum. Periodic acid?Schiff (PAS) staining showed scattered hyphae in the stratum corneum. The internal transcribe spacer(ITS)sequence of the isolated fungus showed 99%consistency with that of Arthrinium phaeospermum. The patient was diagnosed with cutaneous Arthrinium phaeospermum infection, and treated with oral itraconazole capsules 200 mg/d for 16 days. One month later, follow?up showed satisfactory outcomes.