This epidemiological survey was a retrospective study on three nodes during the past three decades on fungal infections representing the China, including Taiwan. Owing to rare publications reporting on dynamic epidemiological trends in the pathogen epidemiology in China, we surveyed the isolation rates and pathogenic fungi from 8 representative districts in China using uniform identification with uniform methodology. The pathogenic fungi isolation rates and species obtained from 1986 (n=9,096), 1996 (n=19,009), and 2006 (n=33,022) suggested that Trichophyton rubrum was the commonest organism cultured in 1980s (45.4%) and 1990s (34.5%), but Candida albicans increased significantly and reaching to its peak (26.9%) in 2006s' survey, and has become the most common isolate of fungal infections in China currently. In addition, Candida glabrata became the most common non-albicans species of Candida in 2006s' survey. At the same time, the incidence of molds also gradually increased. According to comparative analysis of the results of these three surveys, we found apparent differences in the isolation rates of different pathogenic fungi and the forefront 10 species in China varied significantly, and the dermatophytes decreased markedly, while yeasts, especially the Candida species and the molds, increased gradually during the past three decades. Less dermatophytic infections may suggest better access to healthcare or increase in Candida species indicated higher incidence of hospital acquired infections.
A urease-negative serotype A strain of Cryptococcus neoformans (B-4587) was isolated from the cerebrospinal fluid of an immunocompetent patient with a central nervous system infection. The URE1 gene encoding urease failed to complement the mutant phenotype. Urease-positive clones of B-4587 obtained by complementing with a genomic library of strain H99 harboured an episomal plasmid containing DNA inserts with homology to the sudA gene of Aspergillus nidulans. The gene harboured by these plasmids was named URE2 since it enabled the transformants to grow on media containing urea as the sole nitrogen source while the transformants with an empty vector failed to grow. Transformation of strain B-4587 with a plasmid construct containing a truncated version of the URE2 gene failed to complement the urease-negative phenotype. Disruption of the native URE2 gene in a wild-type serotype A strain H99 and a serotype D strain LP1 of C. neoformans resulted in the inability of the strains to grow on media containing urea as the sole nitrogen source, suggesting that the URE2 gene product is involved in the utilization of urea by the organism. Virulence in mice of the urease-negative isolate B-4587, the urease-positive transformants containing the wild-type copy of the URE2 gene, and the urease-negative vector-only transformants was comparable to that of the H99 strain of C. neoformans regardless of the infection route. Virulence of the URE2 disruption stain of H99 was slightly reduced compared to the wildtype strain in the intravenous model but was significantly attenuated in the inhalation model. These results indicate that the importance of urease activity in pathogenicity varies depending on the strains of C. neoformans used and/or the route of infection. Furthermore, this study shows that complementation cloning can serve as a useful tool to functionally identify genes such as URE2 that have otherwise been annotated as hypothetical proteins in genomic databases.
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