下肢深静脉血栓形成继发溃疡性皮肤曲霉菌病1例

报道1例下肢深静脉血栓形成患者继发黄曲霉引起的溃疡性皮肤曲霉病。患者男性,41岁,左小腿胫前外伤后溃烂、流脓、结痂5个月。组织病理检查见较多分隔菌丝,菌丝呈Y型分支。结合真菌镜检、培养,扫描电镜及DNA序列分析鉴定为黄曲霉。给予口服伊曲康唑胶囊及外用0．2％两性霉素B溶液治疗13周,患者皮损明显好转。     

Cutaneous and pulmonary aspergillosis in rabbits.

Cutaneous and pulmonary aspergillosis was diagnosed in rabbits. Dichotomous branching, septate hyphae typical of Aspergillus spp were observed in cystic hair follicles. Atypical, short, knobby mycelia with radiating projections were seen in nodular lesions in the lung.     

声带曲菌病6例报告

目的探讨声带曲菌病的实验诊断和治疗方法。方法分析我院门诊2007年3月～2008年9月间6例声带曲菌病患者的临床资料,总结经验教训。结果本文6例门诊患者声带活检,标本分别作真菌检验和病理检验。真菌直接镜检阳性1例,培养有4例烟曲菌生长和2例黄曲菌生长;病理检验均显示声带坏死组织中大量中性粒细胞、浆细胞及淋巴细胞浸润,4例可见大量菌丝,呈45。分支透明有隔菌丝。嘱患者禁声3d,停用抗生素。予伊曲康唑胶囊0．2g,口服2次／d,14d后减量为0．1g,口服1次／d,7d后停药。患者服药3～4d后能够发声,10d后声音嘶哑症状消失。1—3个月后复查电子喉镜均显示：假膜消失,声带光滑,闭合正常。随访1个月-1年半,6例声音完全恢复正常,声带光滑,声带无充血现象,无复发。结论治疗声带曲菌病应要求患者禁声并停用抗生素,同时予抗真菌药物治疗至关重要。     

Value of antigen and antibody detection,and blood evaluation parameters in diagnosis of avian invasive aspergillosis

The applicability of ELISA detection of circulating Aspergillus spp. antigen (Ag) and systemic antibody (Ab) of IgG class, and the blood parameter values were evaluated for diagnosis of invasive aspergillosis in Aspergillus spp.-challenged Peking ducks (Anas platyrhynchos). The protective role of Aspergillus spp. IgG was evaluated in Cape shelducks (Tadorna cana) immunized with Aspergillus spp. Ag. Challenged but non-immunized A. platyrhynchos developed invasive aspergillosis on day 21 as demonstrated histopathologically by the presence of fungal microgranuloma in air sacs and lung tissue, with serum antigenemia fluctuating from 65 to 270 ng of 55-kD basic protein Ag per ml. Immunized A. platyrhynchos did not demonstrate Aspergillus spp. serum antigenemia but did show rare histopathological changes in some air sacs associated with fungal inflammation. Although the differences between immunized and non-immunized T. cana in blood evaluation parameters did not differ significantly, immunized birds mounted high Aspergillus spp.-specific IgG titer. There was no correlation between the blood parameter values and post-immunization timepoints in T. cana and in A. platyrhynchos. Intramuscular immunization with Aspergillus spp. mycelial phase cultures Ag provided protection against the pathogens. The lack of relations between blood parameter values and increasing Aspergillus spp. IgG titers (in T. cana and A. platyrhynchos) indicate low applicability of these parameters in evaluation of a bird Aspergillus spp. status. Detection of circulating 55-kDa Aspergillus spp. Ag has high early predictive values for invasive aspergillosis in birds. This revised version was published online in August 2006 with corrections to the Cover Date.     

我国首例肌曲霉病及其试验研究

目的报道国内首例肌曲霉病,对所分离的黄曲霉进行形态、药敏、rDNA序列分析研究。方法对患者病变肌组织行B超和病理检查;将分离菌株接种于察氏培养基观察形态学特征;微量法测定MIC值;酚氯仿法提取DNA,扩增ITS间区序列并进行多序列分析。结果B超示左腓肠肌不规则回声,病理示肌细胞变性、坏死,肌组织内见分枝分隔菌丝。表型及基因型特征为黄曲霉;分离菌株对多种抗真菌药物耐药;多基因序列分析显示所分离菌株与标准株及自然和临床分离株具有100%的相似性。给予特比萘芬治疗3个月后痊愈。结论黄曲霉可致肌肉感染;药敏试验对临床治疗具有指导意义;环境中普遍存在的真菌可引起免疫低下患者致死性感染。     

A nonredundant role for plasmacytoid dendritic cells in host defense against the human fungal pathogen Aspergillus fumigatus

While plasmacytoid dendritic cells (pDCs), a natural type I interferon (IFN)-producing cell type, are regarded as critical for innate immunity to viruses, their role in defense against fungal infections remains unknown. We examined the interactions of pDCs with hyphae of the invasive human fungal pathogen Aspergillus fumigatus. Human pDCs spread over hyphae and inhibited their growth. Antifungal activity was retained in pDC lysates, did not require direct fungal contact, and was partially reversed by zinc. Incubation with hyphae resulted in pDC cytotoxicity, partly due to fungal gliotoxin secretion. Following hyphal stimulation, pDCs released proinflammatory cytokines via a TLR9-independent mechanism. Pulmonary challenge of mice with A. fumigatus resulted in a substantial influx of pDCs into lungs, and pDC-depleted mice were hypersusceptible to invasive aspergillosis. These data demonstrate the antifungal activity of pDCs against A. fumigatus and establish their nonredundant role in host defenses against invasive aspergillosis in vivo.     

Granulomatous Invasive Aspergillosis of Paranasal Sinuses Masquerading as Actinomycosis and Review of Published Literature

Cutaneous aspergillosis is a common systemic mycosis affecting immunosuppressed patients. Here, we describe a novel morphological type of cutaneous aspergillosis in a young immunocompetent woman who presented with a chronic history of multiple nodules and discharging sinuses over left side of the face, mimicking cervicofacial actinomycosis. Skin biopsy showed granulomatous inflammation, and of septate fungal hyphae with acute-angled branching, morphologically resembling Aspergillus. This was confirmed on fungal culture as Aspergillus flavus.     

Disseminated Aspergillus terreus infection in a caged pigeon

Disseminated aspergillosis due to Aspergillus terreus was diagnosed in a young pigeon kept by a bird fancier. The fungus was isolated in heavy growth from the infected tissues on Sabouraud medium at 37 °C. Microscopic examination of the squeeze preparation of the air sacs revealed the presence of typical conidiophores of an Aspergillus sp. Histologically, branched, septate hyphae morphologically indistinguishable from Aspergillus were detected in the air sacs and lungs. The clinical, mycological and pathological findings are discussed. The role of A. terreus as an opportunistic pathogen should be studied in various clinical conditions of mammals and avians.     

Use of Mycological, nested PCR, and Real-time PCR Methods on BAL Fluids for Detection of Aspergillus fumigatus and A. flavus in Solid Organ Transplant Recipients

Invasive aspergillosis continues to be a significant cause of morbidity and mortality in solid organ transplant (SOT) recipients. A reliable and early diagnostic method is needed to improve survival. In this study, four methods direct microscopy, culture, nested PCR on internal transcribed spacer region, and TaqMan real-time PCR targeted β-tubulin gene were examined for the detection of Aspergillus fumigatus and A. flavus in sixty-four bronchoalveolar lavage (BAL) fluids that were obtained from SOT recipients. Direct examination with 20 % KOH (potassium hydroxide) and culture on mycological media were also performed. Of the 64 samples, seven (10.9 %) were positive in direct examination (five with septate hyphae and two with aseptate hyphae), and 15 (23 %) had positive culture including five A. flavus, four A. niger, two Penicillium spp., two Rhizopus spp., one Fusarium spp. and one mixed A. flavus/A. niger. Twenty five (39 %) samples had positive nested PCR with A. flavus and 6 (9.4 %) with A. fumigatus-specific primers. Only eight (12.5 %) had positive real-time PCR for A. flavus and nine (14 %) for A. fumigatus. The incidence of aspergillosis in these patients included proven invasive pulmonary aspergillosis (IPA) in two (3 %), probable IPA in 14 (22 %), possible IPA in 38 (59 %), and not IPA in 10 (16 %). A. flavus was the most common cause of pulmonary aspergillosis (PA) in the study. The results suggest that because nested PCR is too sensitive it may increase the number of false-positive results and is not recommended for BAL samples for diagnosis of PA. Although further studies with significant number of proved positive/negative standard BAL samples are necessary for better evaluation, the novel multiplex real-time PCR developed in the study could be promising as a valid diagnostic method for IPA.     

肝移植术后曲霉的气道定植与侵袭性曲霉病的关系

目的评价肝移植术后曲霉气道的定植及其发生侵袭性感染的风险。方法回顾2003—2004年南京八一医院和上海长征医院56例肝移植患者进行的连续组织学检查和曲霉培养结果。结果24例患者被分离出曲霉,2例发生侵袭性烟曲霉感染。这2例患者在移植后6个月内都有烟曲霉的定植,并且都死亡,占所有移植后死亡的29％。无曲霉定植者都未发生侵袭性曲霉感染。结论肝移植术后的侵袭性曲霉感染较为少见,但常致死,曲霉定植常见但为一过性,由于气管侵袭性曲霉感染只发生在有烟曲霉定植的移植后6个月内的患者,所以在此期间进行预防性的治疗能否有效降低侵袭性曲霉感染值得研究。     

Complement induction and complement evasion in patients with cerebral aspergillosis

Cerebral aspergillosis is a mostly lethal infection of the central nervous system. Former results identified low cerebral complement levels as one cause for insufficient immune reaction. Therefore we studied cerebral complement expression after fungal invasion and investigated putative mechanisms of Aspergillus spp to cope with the complement-induced selection pressure. Brain tissue derived from patients with cerebral aspergillosis or non-infected individuals was analyzed immunohistochemically for complement synthesis. Correlations between expression levels were determined statistically. Increased complement synthesis, a prerequisite for strengthened antifungal potency, was visible in resident astrocytes, neurons, oligodendrocytes as well as in infiltrating macrophages after fungal infection. Surprisingly, microglia, although regarded as major immune cells, only marginally participated in synthesis of most complement proteins. Several evasion mechanisms were found that help the fungus to establish a cerebral infection even in the presence of complement: Fungal hyphae limit the surface deposition of C3 and thus interfere with complement-dependent phagocytosis. Furthermore, the "sealing off" in brain abscesses not only inhibits fungal spreading but also forms protection shields against complement attack. Complement indeed seems to represent an important selection pressure and evokes the development of fungal evasion mechanisms. Counteractions for these evasion processes might represent interesting therapeutic approaches.     

Invasive aspergillosis promotes tumor growth and severity in a tumor-bearing mouse model

Invasive aspergillosis increases in chronic immunosuppressive diseases such as cancer. There is little information about the mechanisms by which Aspergillus infection affects the immune regulation and microenvironment of cancer cells. Hence, this study was aimed at investigating the effect of invasive aspergillosis on immunosurveillance, metastasis, and prognosis of cancer in tumor-bearing mice. After implantation of mouse mammary tumor in BALB/c mice, they were infected with Aspergillus conidia intravenously. For comparison, groups of mice were experimentally infected with Aspergillus conidia or implanted with tumor cells separately. Seven days after Aspergillus infection, the serum levels of tissue inhibitor of metalloproteinase-1 (TIMP-1) were measured by ELISA, and subsequently regulatory T lymphocytes were analyzed by flow cytometry. The survival of animals and mean tumor size were then determined. Our results indicated that tumor sizes in mice increased significantly after infection with Aspergillus conidia. Moreover, invasive aspergillosis enhanced the population of regulatory lymphocytes and level of TIMP-1. This study supports the idea that massive Aspergillus infection could stimulate tumor growth and increases the possibility of a bad prognosis. As a result, treatment of Aspergillus infection could be considered an important issue for efficient cancer therapy.     

Effects of non-24-hour days on reproductive efficacy and embryonic development in mice

ICR female mice were exposed to either 22 (L11, D11) or 26 hour day (L13, D13) light/dark cycles for at least 2 weeks before mating and/or during pregnancy. The mating rates of these animals decreased considerably. When pregnant females were examined at gestation days 12.0 or 17.5, resorption rates were increased, the embryos weighed less, and development was retarded in the experimental groups with preconceptional exposure to non-24-hour days. We speculate that in mice maternal and paternal pre- and periconceptional environment of daily light/dark cycles is important for normal reproductive efficacy and normal embryonic development during pregnancy.     

Polarized response of endothelial cells to invasion by Aspergillus fumigatus

Hyphal invasion of blood vessels is a prominent feature of invasive aspergillosis. During invasive pulmonary aspergillosis, Aspergillus fumigatus hyphae invade the abluminal endothelial cell surface, whereas they invade the luminal endothelial cell surface during haematogenous dissemination. We investigated the endothelial cell response to abluminal and luminal infection with A. fumigatus hyphae in vitro . We found that these hyphae invaded the abluminal endothelial cell surface without inducing the formation of endothelial cell pseudopods. Also, the internalized hyphae were surrounded by a loose network of microfilaments. In contrast, A. fumigatus hyphae invaded the luminal endothelial cell surface by inducing by the formation of endothelial cell pseudopods. These endocytosed hyphae were surrounded by a tight network of microfilaments. Abluminal infection induced greater E-selectin, IL-8, tissue factor and TNF-α gene expression, but less endothelial cell damage than did luminal infection. Endothelial cell stimulation by infection of either surface was mediated by endothelial cell-derived TNF-α, and was not influenced by gliotoxin secreted by A. fumigatus . These differences in the endothelial cell response to abluminal versus luminal infection may contribute to differences in the pathogenesis of invasive versus haematogenously disseminated aspergillosis.     

Native valve Aspergillus fumigatus endocarditis with blood culture positive and negative for galactomannan antigen. Case report and literature review

Native valve endocarditis caused by Aspergillus spp. is an uncommon disease with a high mortality rate. Generally, Aspergillus is isolated from affected valve in post-mortem or biopsy specimens. However, its isolation from blood cultures is exceedingly rare. We report a case of fungal endocarditis in a native mitral valve with the isolation of Aspergillus fumigatus both in valve vegetation and in blood culture bottles. The patient underwent valve replacement and antifungal treatment with voriconazole and caspofungin, but he died on post-operative day 45 with disseminated aspergillosis confirmed by necropsy. Paradoxically, galactomannan antigen detection in serum was negative. This is the third case of Aspergillus endocarditis with positive blood culture reported in the literature.     

Comparative studies of prostaglandins F2α and E2 in late cyclic and early pregnant sheep: synthesis by endometrium and conceptus effects of indomethacin treatment on establishment of pregnancy

Endometrial concentrations of prostaglandins F₂α (PGF₂α) and E₂ (PGE₂) were measured by specific radioimmunoassay in sheep, on day 14 of estrous cycle or pregnancy, during luteolysis (Day 16 of the cycle), and after implantation (Day 23 of pregnancy) : concentrations observed on day 14 of cycle and pregnancy were similar. During luteolysis, on day 16 of cycle, a consistent drop was noticed. If luteal regression did not occur, as a consequence of the presence of an embryo, endometrial concentrations of PGF₂α on day 23, were twice those of day 14, and PGE₂ remained unchanged. 2 hour incubations of endometrial caruncular tissue from 14 days cyclic or pregnant ewes resulted in de novo synthesis of PG which could be increased by Arachidonic Acid and inhibited by Indomethacin; during the first 30 min of incubation, the PGF₂α synthesis was comparable for both endometrial tissues, whereas PGE₂ synthesis was twice as great in pregnant endometrium. Fourteen and 23 day conceptuses had high PGF₂α and PGE₂ concentrations which were not due to maternal PG sequestration : PG synthesis which could be inhibited by Indomethacin was observed in incubated 14 day old embryos. Treatment of pregnant ewes from day 7 to day 22 after mating, either with Indomethacin (300 mg s.c. daily) or with Acetylsalicylic Acid (1 g I.V. daily) resulted in a sharp diminution of endometrial PG concentration and release, with no apparent effect on the establishment of pregnancy. These results tend to ascribe a less important role to PG during early pregnancy in sheep as compared with rodents, in terms of embryonic growth and implantation.     

Comparative studies of prostaglandins F2α and E2 in late cyclic and early pregnant sheep: In vitro synthesis by endometrium and conceptus effects of in vivo indomethacin treatment on establishment of pregnancy

Endometrial concentrations of prostaglandins F₂α (PGF₂α) and E₂ (PGE₂) were measured by specific radioimmunoassay in sheep, on day 14 of estrous cycle or pregnancy, during luteolysis (Day 16 of the cycle), and after implantation (Day 23 of pregnancy) : concentrations observed on day 14 of cycle and pregnancy were similar. During luteolysis, on day 16 of cycle, a consistent drop was noticed. If luteal regression did not occur, as a consequence of the presence of an embryo, endometrial concentrations of PGF₂α on day 23, were twice those of day 14, and PGE₂ remained unchanged. $\text{In vitro}$ 2 hour incubations of endometrial caruncular tissue from 14 days cyclic or pregnant ewes resulted in de novo synthesis of PG which could be increased by Arachidonic Acid and inhibited by Indomethacin; during the first 30 min of incubation, the PGF₂α synthesis was comparable for both endometrial tissues, whereas PGE₂ synthesis was twice as great in pregnant endometrium. Fourteen and 23 day conceptuses had high PGF₂α and PGE₂ concentrations which were not due to maternal PG sequestration : $\text{de novo}$ PG synthesis which could be inhibited by Indomethacin was observed in incubated 14 day old embryos. Treatment of pregnant ewes from day 7 to day 22 after mating, either with Indomethacin (300 mg s.c. daily) or with Acetylsalicylic Acid (1 g I.V. daily) resulted in a sharp diminution of endometrial PG concentration and release, with no apparent effect on the establishment of pregnancy. These results tend to ascribe a less important role to PG during early pregnancy in sheep as compared with rodents, in terms of embryonic growth and implantation.     

Taxonomy and identification of the species involved in nosocomial aspergillosis

Although Aspergillus fumigatus is the most common etiological agent of invasive aspergillosis, other Aspergillus spp. such as A . flavus, A. niger, A. terreus and A. nidulans (Emericella nidulans) among others, have been also implicated. In this article, the taxonomy of the genus Aspergillus and the characteristics of the species most frequently isolated from patients with nosocomial aspergillosis are reviewed.     

Selective expression of a major allergen and cytotoxin, Asp f I, in Aspergillus fumigatus. Implications for the immunopathogenesis of Aspergillus-related diseases.

Asp f I is a major 18-kDa Aspergillus fumigatus allergen and a member of the mitogillin family of cytotoxins. The nucleotide sequence of the Asp f I gene was determined by sequencing polymerase chain reaction products amplified from A. fumigatus spore DNA. The entire 678-bp DNA includes an 81-bp leader sequence, preceding the N-terminal alanine codon, a 52-bp intron, and a 444-bp open reading frame, encoding a 149-amino acid protein (M(r) 16,899), which is 99% homologous to mitogillin from Aspergillus restrictus. A mAb-based ELISA was used to compare Asp f I levels in spores, mycelia, and culture filtrate, and to determine the kinetics of allergen production. Disrupted hyphae or spore extracts had a 1000-fold lower level of Asp f I than culture filtrate, suggesting that germination of spores and growth of the fungus are essential for allergen production. Asp f I levels in A. fumigatus and A. restrictus peaked at day 3 (0.87 to 12.1 micrograms/ml), however, the allergen was not detected in Aspergillus flavus, Aspergillus niger, Aspergillus terreus, and Aspergillus nidulans cultures (< 1.5 ng/ml) on either days 3 or 8. Northern analysis confirmed that Asp f I mRNA was detected only in A. fumigatus and A. restrictus, but not in the other four Aspergillus spp. Asp f I-specific DNA was generated after polymerase chain reaction amplification of genomic mycelial DNA obtained from A. fumigatus and A. restrictus, but not from the other Aspergillus spp. The results show that Asp f I is selectively expressed in A. fumigatus, and suggest that this cytotoxin could be a specific virulence factor for A. fumigatus.     

Early pregnancy diagnosis by the ultrasonographical device and observation of fatal growth in cynomolgus monkeys (Macaca fascicularis)

Usefulness of the ultrasonographical device (USD) for early diagnosis of pregnancy and for observation of fetal growth was examined in the cynomolgus monkey (Macaca fascicularis). Firstly, 50 females, each of which was mated for 24 hours in an individual cage, were used. They were diagnosed for pregnancy by the use of USD 15 to 29 days and 35 days after mating. Then, 20 of them were judged to be pregnant. The early pregnancy diagnosis was based on the detection of gestational sac (GS) in the uterus. The GS was detected 20.0 +/- 1.8 days after mating. The duration from the day on which GS was first detected to the day on which the heart beat of embryo (HBE) was detected averaged 5.6 +/- 1.5 days. In 17 of these 20 pregnancies, GS was detected 17 to 23 days after mating. As regards the remaining three pregnant cases, GS and HBE were simultaneously detected on the 28th day of pregnancy in two cases, and on the 35th day in one case. Secondly, 225 females who were mated by the routine 3 day-mating system were used. Pregnancy diagnosis by USD was conducted on the 21st, 28th and 35th day after mating. Ninety-six of the 225 females were judged to be pregnant. In 33 (34.1%) of the 96 pregnant cases, GS was detected on the 21st day of pregnancy. In 57 females (59.4%), GS was detected on the 35th day. Furthermore, it was able to judge conditions of fetal growth by measuring the size of GS and biparietal diameter.(ABSTRACT TRUNCATED AT 250 WORDS)