Two new 12-membered macrolides from the endophytic fungal strain Cladosprium colocasiae A801 of Callistemon viminalis

Two new polyketide metabolites, the 12-membered macrolides 4-hydroxy-12-methyloxacyclododecane-2,5,6-trione (1) and 12-methyloxacyclododecane-2,5,6-trione (2), were isolated from the endophytic fungal strain Cladosprium colocasiae A801 of the plant Callistemon viminalis, together with five known derivatives. Their structures were fully characterized by means of detailed spectroscopic analysis for new structures, and in comparison with published data for known compounds. The antibacterial, cytotoxic, and α-glucosidase inhibitory activities of the new compounds 1 and 2 were evaluated.     

痔体黏膜发现白色念珠菌寄生

为探讨痔的病因,将26例患者痔体送病理组织检查,PAS染色发现9例有紫红色菌体寄生,据其形态判断是白色念珠菌,试管沙氏培养基培养见奶白色酵母样菌落,转种于科玛嘉培养基培养见菌落呈翠绿色,诊断为白色念珠菌。由此推断痔上皮增生、角化,黏膜大量炎细胞浸润,间质水肿、出血,黏膜下静脉强烈炎性变,是因白色念珠菌感染引起的。     

Effects of Fungi and Eosinophils on Mucin Gene Expression in Rhinovirus-Infected Nasal Epithelial Cells

Purpose

Fungi, rhinoviruses (RVs), and eosinophils are associated with upper respiratory diseases. We evaluated the effects of fungal stimulation and eosinophil co-culture on the expression of mucin genes in RV-infected nasal polyp epithelial cells.

Methods

Nasal polyp epithelial cells were obtained from chronic rhinosinusitis patients. Cultured epithelial cells were stimulated with Alternaria and Aspergillus with or without RV-16 infection. The epithelial cells were co-cultured with eosinophils for 16 h. MUC4, MUC5AC, MUC5B, and MUC8 mRNA expressions in the epithelial cells were quantified using real-time RT-PCR. To determine the underlying mechanism, nuclear factor-κB (NF-κB), activator protein-1 (AP-1), and mitogen-activated protein kinase (MAPK) inhibitors were used to inhibit mucin gene expression.

Results

Fungi and RV-16 induced mucin gene expression in nasal polyp epithelial cells. However, there was no synergistic increase in mucin gene expression, with the exception of MUC4 mRNA expression stimulated by 25 µg/mL Aspergillus. When RV-16-infected epithelial cells were stimulated with fungi and then co-cultured with eosinophils, MUC4, MUC5B, and MUC8 mRNA expressions increased. Mucin gene expression was inhibited by NF-κB inhibitors.

Conclusions

RV-16, airborne fungi, and eosinophils may exacerbate the inflammatory process in nasal mucosal diseases by enhancing mucin gene expression.     

丹东近海真菌Aspergillus pseudoglaucus次级代谢产物的研究

目的 研究海洋来源真菌假灰绿曲霉Aspergillus pseudog1aucus次级代谢产物的化学成分及其抗氧化活性.方法 摇床发酵后,采用硅胶柱层析、LH20凝胶色谱和半制备高效液相色谱等方法,对假灰绿曲霉发酵总浸膏进行分离纯化;利用核磁共振(NMR)、质谱(MS)、旋光和单晶X-ray衍射技术结合文献报道数据确定...     

198例酵母样真菌感染及其耐药性分析

目的了解院内患者的真菌感染情况,对其耐药性进行分析,以指导临床合理用药。方法用念珠菌显色平板进行初步鉴定,再使用ID32C和ATB FUNGUS3分别进行鉴定和药敏试验,在全自动鉴定与药敏测试仪（ATB ExPression）上读板。结果送栓1045例标本,检出酵母样真菌198株,院内感染真菌分离率为18．9％（198／1045）。其中白色念珠菌140株,热带念珠菌25株,清酒念珠菌13株,光滑念珠菌8株,克柔念珠菌4株,其他念珠菌8株,分别占检出真菌的70．7％、12．6％、6．6％、4．0％、2．0％、4．0％。结论以白色念珠菌感染为主,热带念珠菌和清酒念珠菌其次,3种念珠菌占总分离真菌的89．9％（178／198）。5种抗真菌药物的耐药率以5－氟胞嘧啶和两性霉素B较低,氟康唑、伊曲康唑和伏立康唑较高。     

毛花猕猴桃可培养内生真菌菌群结构、多样性及生物活性分析

目的 探讨毛花猕猴桃Actinidia eriantha可培养内生真菌的菌群结构、多样性、分布规律及其活性,为内生真菌资源利用提供依据。方法 对毛花猕猴桃3个部位（根、茎、叶）内生真菌进行分离、纯化与ITS分子鉴定,分析其菌群结构、多样性与分布规律;采用拟南芥共培养、病原菌平板对峙的方法,对内生真菌进行促生与病原菌拮抗活性筛选。结果 共分离得到2 446株内生真菌,分为156个形态型,ITS分子鉴定为49个分类单位,归属于真菌界3门5纲10目14科18属,其中子囊菌门真菌数量最多,占比94.89%;根、茎、叶共有菌30种,其中Trichoderma spirale是根的特有种,Bionectria ochroleuca是茎的特有种;Shannon-Wiener（H’）以茎多样性指数最高,Simpson指数（D）以叶多样性指数最高;筛选得到显著促拟南芥生长活性的内生真菌AE16,鉴定为Clonostachys sp.,其共生拟南芥鲜质量增长率为（37.13±10.35）%;具有显著抑菌活性的内生真菌11株（木霉属10株,白僵菌属1株）,其中AE175（Trichodermasp.）抑制效果最好,相对抑菌率为（87.59±9.36）%。结论 畲药毛花猕猴桃内生真菌资源丰富,筛选得到的活性菌株为毛花猕猴桃内生真菌资源开发和利用提供依据。     

婴儿院内肺部真菌感染的危险因素

目的 探讨新生儿科住院患儿院内肺部真菌感染(NIPFI)的流行病学及其危险因素.方法 选取2006年7月1日-2008年6月30日在广东省人民医院新生儿科住院的患儿中符合院内深部真菌感染(NIFI)诊断标准的婴儿[日龄(77±61) d]共68例.收集患儿一般资料、感染前诊疗经过、标本类型等,并对NIPFI及其他NIFI患儿各项数据进行比较.应用SPSS 13.0软件进行统计学分析.结果 住院患儿NIFI发病率为3.31%(68/2 052例),其中NIPFI发生率为1.36%(28/2 052例).NIPFI患儿总住院时间中位数为15.50 d;心血管疾病患病率为94.6%,感染真菌前广谱抗生素的使用率为96.4%,抗生素的联合应用占64.3%;78.6%的患儿接受了心脏手术;机械通气率为82.1%;中心静脉置管为67.9%;留置胃管率为96.4%;激素使用者占78.6%.NIPFI患儿机械通气率较其他NIFI患儿高(χ2=7.32,P=0.010);余因素在2组患儿间比较差异均无统计学意义.多元回归分析显示机械通气与NIPFI有显著相关性(OR=0.229,95%CI 0.072～0.725,P=0.012).结论 在NIFI的各种危险因素中,机械通气是NIPFI的危险因素.     

Use of <Emphasis Type="Italic">MET3</Emphasis> promoters for regulated gene expression in <Emphasis Type="Italic">Ashbya gossypii</Emphasis>

A central tool for gene function analysis is the construction mutant strains. This can be done conveniently in A. gossypii using PCR-based tools. The deletion of essential genes can be performed since initial transformants are sheltered in a heterokaryotic mycelium, which contains nuclei with both wild type and mutant alleles. The analysis of mutant phenotypes in A. gossypii is regularly started by germinating spores, which contain only one nucleus. Thus, selection can be used to identify mutant germ cells and germlings. However, such an analysis yields only mutant mycelia if the deleted gene is not essential. We describe the use of the regulatable Saccharomyces cerevisiae and A. gossypii MET3 promoters as novel tools to regulate gene expression in A. gossypii. Conditional expression was tested using GFP and lacZ-reporter genes. Regulation of MET3 promoters was found to be dependent on methionine but not on cysteine and down-regulation to about 1/10 of the initial expression levels was achieved. We used the A. gossypii WAL1 and CYK1 genes as models to demonstrate that MET3 promoters could regulate the expression of these genes and reveal their mutant phenotypes depending on the presence or absence of methionine. Finally, we show that the AgMET3 promoter contains two Cpf1-binding sites and that AgCPF1 can complement the S. cerevisiae cpf1 methionine auxotrophy.