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Objective To establish a rat model of Cryptococcus neoformans meningitis. Methods Sprague-Dawley rats were divided into four groups (group A to D). The load of Cryptococcus neoformans was inoculated by intracisternal injection to the animals in Group A (1×105 cfu), Group B (1×106 cfu), Group C (1×107 cfu), and 0.9％ NaCl solution to Group D. The rats were observed after inoculation for their clinical symptoms. On day 14 and day 21 after inoculation, cerebrospinal fluid was sampled and cultured for counting of bacterial colonies. The 28-day mortality of the animals was calculated. Results All the animals in group A (1×105 cfu) survived without any apparent clinical symptoms, and associated with decreasing bacteria load. The animals in group B (1×106 cfu) had mild symptoms associated with low mortality rate and slightly increased bacterial load. The animals in group C (1×107 cfu) showed a lot of symptoms and associated with high mortality rate and significantly increased bacteria load. All the animals in group D (0.9％ NaCl solution) survived with normal activity. No bacterial colony was cultured from the cerebrospinal fluid. Conclusions Intracisternal injection of 1×107 cfu Cryptococcus neoformans to rats could cause apparent clinical symptoms of meningitis. The 28-day mortality rate of such a rat model of Cryptococcus neoformans meningitis is greater than 80％. An ideal rat model of Cryptococcus neoformans meningitis is established successfully.
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Objective To analyze the differential expression of microRNA-146a (miR-146a) in monocyte-macrophage cell line (THP-1 cells) after induction by Cryptococcus neoformans (C.neoformans,reference strain WM148) or Cryptococcus gattii (C.gattii,reference strain R265),and investigate the mechanism of miR-146a in regulating the inflammatory response of cryptococcal meningitis.Methods The cultured THP-1 cells were divided into two groups to be induced by C.neoformans or C.gattii,respectively.THP-1 cells were induced with inactivated WN148 (or R265) strains at multiplicity of infection (MOI) of 5 in all experiments.The supematant and the cell pellet were collected separately after incubation.The expression of miR-146a was measured by real-time quantitative PCR (qRT-PCR) technique.The levels of TNF-u and IL-6 release were assayed by ELISA.Results The expression of miR-146a increased significantly in the C.neoformans induction group compared to 0 h.It reached peak at 3 h (P＜0.01),and then declined gradually.The level of TNF-α increased in supematant and reached peak at 12 h.The expression of IL-6 did not change significantly at each time point.The expression of miR-146a and TNF-α increased gradually and reached peak at 12 h in the C.gattii induction group (P ＜0.01),but the change did not reach statistical significance at 3 h,6 h time points.The expression of IL-6 gradually increased,and reached peak at 12 h time point.Conclusions Following stimulation with C.neoformans or C.gattii,the expression ofmiR-146a in THP-1 cells showed different patterns over time.The expression levels of TNF-α and IL-6 showed different patterns.These findings suggest that there may be different regulatory mechanisms in the THP-1 cells-associated inflammatory response after stimulation by inactivated C.neoformans and C.gattii strains.
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Objective To review the clinical features and diagnosis of syphilitic aortic aneurysm .Methods A case of syphilitic aortic aneurysm was reported and the relevant literatures were reviewed .Results The patient showed progressive hoarse and heart murmurs and was diagnosed by computed tomographic angiography of aorta and syphilis serology test .In the era of antibiotics ,syphilitic aortic aneurysm is a very rare and dangerous case with covert onset ,various manifestations and high mortality .Conclusions The prevalence of late cardiovascular syphilis is increasing with the increase of early-onset syphilis and HIV epidemic .Syphilis serologic test should be available for the young patients with aortic aneurysm even though the risk factors of coronary heart disease are not present .Medical and surgical treatments for syphilis should be provided for the patients who are clinically suspected of syphilitic aortic aneurysm .
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Objective To determine the genetic subtypes and susceptible population of Cryptoccocus neoformans in China. Methods We analyzed 129 strains of Cryptococcus neoformans isolated during the period from 1980 through 2006 in 18 Prov-inces and municipalities of central and eastern China using epidemiological method,PCR fingerprint,and multilocus sequence typing (MLST).Results About 71.5% (91/129)of the clinical strains were isolated from the patients without any recognizable immunodeficiency.Only 8.5% (11/129)of the strains were from AIDS patients.The remaining 20.9% (27/129)were from patients with underlying diseases other than HIV infection.Furthermore,PCR fingerprinting and MLST showed that serotype A strains exhibited a unique VNⅠsubtype,which was distinguishable from the reference VNⅠmolecular types.For convenience we named this unique genotype as VNⅠc.Conclusions People without any recognizable immunodeficiency are the main susceptible population of the 129 clinical strains of Cryptococcus neoformans ,which contrasts with the reports from other countries.The different genetic subtypes of Cryptococcus neoformans between China and other countries may contribute to such a difference.
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ObjectiveTo compare the efficacy and tolerability of 1-week 1％ terbinafine hydrochloride cream, 1- and 4-week 2％ miconazole nitrate cream in the treatment of interdigital tinea pedis, and to observe the relapse in patients treated with these regimens. MethodsA multi-center, randomized, double-blind and parallel group study was conducted. By using a stratified randomization protocol, patients were divided into 3 groups to apply terbinafine cream twice daily for 1 week and inert cream(placebo) for the next 3 weeks (1week terbinafine group), miconazole cream twice daily for 1 week and inert cream(placebo) for the next 3 weeks (1-week miconazole group), and miconazole cream twice daily for 4 weeks (4-week miconazole group),respectively. Clinical and mycological assessment was made on week 1, 3, 4, 6, 9 and 12 after the initiation of treatment. ResultsA total of 152 patients with positive baseline mycological culture were eligible for the efficacy analysis. After 4-week treatment, the mycological cure rates were 94.7％, 87.8％ and 82.6％, global effective rates 89.5％, 81.6％ and 63.0％, respectively for the 1-week terbinafine group, 4-week miconazole group and 1-week miconazole group. On week 12, the mycological relapse rates in 1-week terbinafine, 4-week miconazole and 1-week miconazole group were 13％, 14％ and 21％ respectively, and the incidence of adverse reaction was 2.38％, 2.38％ and 3.57％, respectively. ConclusionsAs far as the efficacy and recurrence in patients are concerned, the 1-week terbinafine cream regimen is similar to the 4-week miconazole cream regimen for the treatment of interdigital tinea pedis.
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Objective To clarify the quorum sensing and identify the quorum sensing molecular of Cryptococcus neoformans. Methods The growth of Cryptococcus neoformans TUP1 △ strain at different inoculum size was observed. The culture filtrate of high cell density TUP1△ strain was collected and plated on medium with low density TUP1△ cell, and the growth of low cell density TUP1△ was recorded. Finally, the active molecule in culture filtrate was purified and identified.Results The growth of Cryptococcus neoformans TUP1△ strain was density dependent and the colony could be normally formed only above a specific density. The filtrate from a high cell density TUP1△culture could promote the growth of low cell density TUP1△, and the active molecule in this culture filtrate was identified to be an oligopeptide composed of 11 amino acids pepride, named quorum sensing peptide 1(QSP1). QSP1 was artificially synthesized, which could also promote the colony forming of TUP1△ strain inoculated at low cell density. Conclusion Quorum sensing exists in Cryptococcus neoformans, and the quorum sensing molecular is QSP1.
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Objective To examine mycological profile of eryptococcal meningitis in patients with non-acquired immune deficiency syndrome (AIDS) during treatment and follow-up so that to support clinical therapy. Methods Data of 28 cuhure-confirmed cryptoeoccal meningitis patients with non-AIDS were retrospectively analyzed. Fungat smear, count, culture and latex agglutination test of cerebrospinal fluid (CSF) were done during treatment and follow-up. Initial treatment included intravenous amphotericin B plus oral flucytosine or f;uconazole for at least 6 weeks, and consolidation treatment included oral fluconazole and (or) itraeonazole for at least 2 months. All 28 patients were cured. The data were analyzed by rank-sum test. Results The positive rate of CSF fungal smear was 92.9% before treatment and gradually decreased, and the fungal count was significantly reduced over time after treatment. While fungal smears of some patients were still positive after initial treatment. Fungal growth time in culture was gradually extended, and fungal culture turned to be negative in all patients after 2 weeks of treatment. The positive rate of latex agglutination test of CSF was 100%. Cryptococcal antigen titer decreased steadily after treatment, which was not correlated with the decrease of fungal count. Conclusion Mycological tests of patients with eryptococcal meningitis should be interpreted comprehensively during treatment, and result of each test should be specifically analyzed.
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0.05).PZs of clinical and environmental isolates were0.501?0.049and0.565?0.131,respectively(P
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Objectives：To study the molecular mechanism of Cryptococcus neoformans mutation in the production of melanin.Methods：By comparing wild types(Mel+) and mutants(Mel-),we detected C.neoformans mutant laccase (CNLAC1) gene by PCR.We also cloned gene of the CNLAC1 containing about 450bp in length.E.coli DH5α was the host strain for the plasmid containing the DNA clone.Results：No deletion was found in wild type.However,CNLAC1 deletion (about 450 bp) was present (between 1 715 bp and 2 617 bp).CNLAC1 containing about 450 bp in length was cloned successfully.Conclusions：The present study was helpful in the biochemical and molecular study of CNLAC1 mutation.It might also provide us a target gene for the prognosis and gene therapy of Cryptocosis.
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Objective To explore the mechanism of Cryptococcus neoformans mutant in CNLAC1and investigate the difference of CNLAC1between wild type(Mel+ )and the mutant (Mel- ).Methods The ab-normal PCR DNA product of Cryptococcus neoformans was cloned into sequencing vector PG EM-T for se-quencing.An expression vector was c onstructed for efficient expressio n in E.coli DH 5? .The gene was sequenced.Results The cloned DNA sequence was found to b e different between wild type(Mel+)and the mutant (Mel-)(between 1715bp and 2617bp).The differences were not only in len gth,but also in base sequence.Con-clusion The mechanism of the mutant in CNLAC1might be a change of sequence(between 1715bp and 2617bp),and not a DNA deletion.This finding might provide us a target gene for the prognosis and treatment of cryptococcosis.
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Objective To investigate and report a group of patients with cutaneous nocardiosis occurring in China.Methods The skin lesion of twenty-three patients were examined and the pathogen was identified.The possible cause of this disease was also investigated.Results All the twenty-three patients belon g to a same village in a province.Skin lesions on buttoc ks manifested as nodules and abscesses whereas some other lesions as ulcers and draining sinuses,occurring 2-10mo nths after intramuscular injection.The pathogens were identified as Nocardia asteroides.Conclusion Twenty-three patients of cutaneous nocardiosis were related to intramu scular injection.
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Objective:To study the role of RapIDYeast Plus (RYP) system in identifying common clinical yeasts.Methods: The target strains were cultured and passaged twice on Sabouraud dextrose agar,and were fed to RYP system after 24-48 h incubation at 30℃.Results: One hundred and thirty-nine of the 150 target strains were identified to the level of species correctly,and 8 undetermined strains were confirmed by additional tests.It was found that 3 strains had been incorrectly identified by RYP system.The accuracy of RYP system was calculated as 92% without additional tests and 98% with additional tests.Conclusion: RYP system is suitable for routine tests in clinical microbiological laboratory;it can accurately identify more than 40 kinds of yeasts and yeast-like bacteria in clinical practice.
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The quorum sensing commonly exists in procaryote kingdom,regulating various biologic functions.Recently similar phenomenon was also found in fungi world,which affecting both biofilm formation and dimorphism.In this article,we focus on the recent progress on quorum sensing of pathogenic fungi and discuss the possibility of taking quorum sensing molecule as a potential target for antifungl therapy.