Establishment of a rat model of Cryptococcus neoformans meningitis / 中国感染与化疗杂志

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.

Mechanistic study on the role of microRNA-146a in THP-1 cells-associated inflammatory response induced by Cryptococcus neoformans and Cryptococcus gattii / 中国感染与化疗杂志

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.

Syphilitic aortic aneurysm:one case report and literature review / 中国感染与化疗杂志

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 .

Genotype and clinical analysis of 129 strains of Cryptoccocus neoformans / 中国感染与化疗杂志

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.

Cryptococcosis in China (1985-2010): review of cases from Chinese database.

BACKGROUND: Cryptococcosis is a potential fatal disease, especially in immunocompromised patients. In China, the profile of cryptococcosis is unclear. Therefore, we summarize the epidemiology and therapy of cryptococcosis in china. METHODS: All cases reports about cryptococcosis in China were collected from CBMdisk database (China Biology and Medicine data disc) with key words of cryptococcosis, or cryptococcal infection, or cryptococcus, and case. The features of the cryptococcosis were retrospectively analyzed. RESULTS: There were 1,032 reports about cryptococcosis, including 8,769 cases. Among them, there were 16% patient with AIDS/HIV, and 17% ones without underlying diseases. There were 2,371 cases of CNS infection. Among them of 2,068 cases, the treatment protocols and outcome were clearly described. The percentages of patients who received intrathecal treatment of amphotericin B(AmB), AmB + 5-FC(5-fluorocytosine), AmB + FCZ(fluconazole), and AmB + 5-FU + FCZ in each medication group were 10, 43, 53, and 33%, respectively. The mortalities were significantly lower in the AmB, Amb + 5-FC, AmB + FCZ intrathecal treatment groups compared with their non-intrathecal treatment controls (6% vs. 23%, 25% vs. 35%, 20% vs. 30%, respectively, P < 0.05), but not in the intrathecal AmB + 5-FU + FCZ group (35% vs. 26%, P > 0.05). CONCLUSION: The Chinese cryptococcosis had its own special clinical features, such as more patients without identifiable underlying diseases. Intrathecal injection of amphotericin B was effective treatment method for cryptococcal CNS infection in China.

Cryptococcal meningitis in immunocompetent children.

To describe clinical characteristics, treatment and outcome of cryptococcal meningitis in immunocompetent children. Immunocompetent children with cryptococcal meningitis who attended Changzheng Hospital between 1998 and 2007 were retrospectively reviewed. During the 10 years reviewed, 11 children with cryptococcal meningitis were admitted to Changzheng hospital and identified as immunocompetent. The 11 children had a median age of 7.25 years. Headache (100%), fever (81.8%), nausea or vomiting (63.6%) and visual or hearing damage or loss (36.4%) were the most common symptoms before treatment. There is no evidence for other site infection of cryptococcus although all the cryptococcal antigen titre is high in blood. All the patients received amphotericin B or AmB liposome with 5-flucytosine for at least 6 weeks followed by fluconazole or itraconazole as consolidation treatment for at least 12 weeks. Nine patients were cured mycologically; however, sequela of visual damage was showed in one patient. Cryptococcal meningitis seems to be uncharacteristic of symptoms, and central nervous system may be the only common site for infection. Amphotericin B with 5-flucytosine should be the choice of induction treatment in this group of patients.

Lumbar puncture drainage with intrathecal injection of amphotericin B for control of cryptococcal meningitis.

Cryptococcal meningitis is a disease with high mortality and refractory to intravenous antifungal treatments with agents such as amphotericin B and fluconazole. We investigated lumbar puncture catheter drainage with an intrathecal injection of amphotericin B as a treatment for cryptococcal meningitis. All of the 14 patients enrolled in the treatment group survived with no evidence of relapse during 1-year follow-up. Complications included lumbosacral nerve root irritation in seven patients and urinary retention in seven patients. This study demonstrated that the technique used was effective in controlling the symptoms. The major complications disappeared after discontinuation of intrathecal injection of amphotericin B or with low-dose therapy. Therefore, this technique could be an effective and safe method for the treatment of cryptococcal meningitis.

Comparison of 1-week terbinafime hydrochloride cream, 1- and 4-week miconazole nitrate cream in the treatment of interdigital tinea pedis: a multi-center, randomized and double-blind study / 中华皮肤科杂志

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.

Aspergillosis of biliary tract after liver transplantation: a case report.

We reported a case of aspergillosis presented as cholangitis in a patient after liver transplantation, even with prophylactic use of fluconazole. The patient had multiple predisposing factors, such as leukocytopenia, immunosuppressive drug therapy. He died 2 days after an exploratory laparotomy was carried out. The histopathologic finding of biopsy specimen from biliary tract was positive for Aspergillus. And the pathogen was identified as Aspergillus flavus by mycological culture and PCR. The patient was confirmed as a case of Aspergillosis of biliary tract that was responded poorly to fluconazole. This indicated that azole should have been switched to more effective antifungal agents at earlier stage when the patients responded poorly to the original treatment.

Identification of quorum sensing molecular in Cryptococcus neoformans TUP1△ strain / 中华传染病杂志

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.

Primary cutaneous aspergillosis in a patient with cutaneous T-cell lymphoma.

We present a case of primary cutaneous aspergillosis caused by Aspergillus terreus in a patient with cutaneous T-cell lymphoma. A 41-year-old woman bruised her left shin and presented with a 1 x 2 cm ecchymosis on the upper third of the left shin for over 6 months. Before mycological examination and sequential biopsies, the patient was erroneously treated with antibiotics and local debridement. After final diagnosis of cutaneous aspergillosis caused by A. terreus cutaneous T-cell lymphoma on the basis of mycological, histopathological and immunohistochemical findings, the patient responded well to oral itraconazole and chemotherapy for over 1 month.

Cryptococcal meningitis in an immunocompetent 8-year-old girl.

An immunocompetent 8-year-old girl was presented with cryptococcal meningitis. She was cured clinically and mycologically after 6 weeks of amphotericin B plus flucytocin and 2 months of fluconazole, and no relapse and sequelae were observed. Mycological and clinical profile of the patient during treatment was recorded.

Mycological profile of cryptococcal meningitis in patients with non-acquired immune deficiency syndrome during treatment and follow-up / 中华传染病杂志

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.

Recurrent pulmonary cryptococcosis in a patient with idiopathic CD4 lymphocytopenia.

A case of recurrent cryptococcosis with idiopathic CD4 lymphocytopenia is reported in this article. After an initial cryptococcal infection in the lung, the patient experienced one episode of cryptococcal meningitis and two more episodes of cryptococcal pneumonia within a period of 12 years. Genetic studies revealed that all isolated microbes were identical, indicating that all subsequent episodes were recurrence instead of re-infection.

The Effect of Phospholipase on the Virulence of Cryptococcus neoformans / 中华皮肤科杂志

0.05).PZs of clinical and environmental isolates were0.501?0.049and0.565?0.131,respectively(P

A preliminary study of the molecular mechanism of Cryptococcus neoformans mutation in the production of melanin / 医学研究生学报

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.

CNLAC1Gene Mutation of Cryptococcus neoformans / 中华皮肤科杂志

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.

Primary Cutaneous Nocardiosis Associated with Intramuscular Injection / 中华皮肤科杂志

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.

Clinical application of RapIDYeast Plus system in identifying 150 clinically common yeasts / 第二军医大学学报

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.

The development of quorum sensing phenomenon in pathogenic fungi / 第二军医大学学报

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.