Dectin-1和TLR2/TLR3/TLR4受体在免疫缺陷小鼠侵袭性肺曲霉菌病的表达

目的本研究利用小鼠动物模型,探讨烟曲霉感染免疫缺陷小鼠TLR2/TLR3/TLR4对树突状细胞植物血凝素-1(Dectin-1)表达的影响。通过检测TLRs和CLRs受体在不同免疫状态下的表达,揭示介导侵袭性肺曲霉菌病(IPA)肺损伤的关键受体。方法小鼠随机分成四组,A组为正常对照组;B组为环磷酰胺免疫抑制+未接种烟曲霉菌;C组正常状态小鼠+烟曲霉菌接种;D组为IPA感染组,免疫抑制+烟曲霉菌接种。采用real-time PCR方法进行小鼠肺组织各个时相点的TLR2、TLR3、TLR4、Dectin-1和β-actin m RNA的表达,检测受体间的相互表达调控。结果正常状态感染组(C组N+AF),TLR4/TLR2、Dectin-1 m RNA比正常对照组表达上调;IPA模型组(D组CP+AF),Dectin-1和TLR4、TLR2 m RNA比正常状态感染组表达下调;但TLR3 m RNA 48 h、72 h表达没有明显不同(P>0.05)。病理切片模型组小鼠72 h可见较严重的出血和充血;96 h时肺内有菌丝,肺泡间隔增宽,支气管壁破坏。正常状态感染烟曲霉小鼠72 h可见充血,肺泡弹性纤维破坏。结论成功建立了小鼠IPA动物模型,Dectin-1受体表达下调可能是环磷酰胺免疫抑制引起IPA的机制之一。Dectin-1的表达可能是建立在TLR2/TLR4对烟曲霉早期识别的基础上,TLR3可能起协同作用。     

侵袭性肺曲霉菌病病人的护理

侵袭性肺曲霉菌病（IPA）是怖曲霉菌病中最严重的类型,诊断困难,治疗棘手,除肺部病变外,尚可合并曲霉菌败血症和其他器官受累。近年来由于广谱抗菌药物、糖皮质激素和免疫抑制剂的大量应用、器官移植和肿瘤放、化疗的广泛开展及艾滋病的流行造成IPA的危险因素增加[1],糖尿病造成免疫妥协易继发多种感染,曲霉菌病也不例外,糖尿病应作为IPA的危险因素引起医师的重视[2]。     

侵袭性肺曲霉菌病病人的护理

侵袭性肺曲霉菌病(IPA)是肺曲霉菌病中最严重的类型,诊断困难,治疗棘手,除肺部病变外,尚可合并曲霉菌败血症和其他器官受累.近年来由于广谱抗菌药物、糖皮质激素和免疫抑制剂的大量应用、器官移植和肿瘤放、化疗的广泛开展及艾滋病的流行造成IPA的危险因素增加[1],糖尿病造成免疫妥协易继发多种感染,曲霉菌病也不例外,糖尿病应作为IPA的危险因素引起医师的重视[2].IPA多发生在有严重基础疾病的病人,预后差,病死率达50％～100％[3].这对有效治疗和护理提出了更高要求.我院呼吸科2007年10月-2010年10月收治IPA住院病人11例,经过系统的治疗、采取有效的护理措施,取得满意的疗效.现报道如下.     

侵袭性肺曲霉菌病11例临床分析

目的探讨侵袭性肺曲霉菌病(IPA)的危险因素、临床表现、诊断和治疗特点。方法回顾性分析2006年1月至2010年10月收治的11例IPA的基础疾病、危险因素、临床表现、实验室及影像学特征、治疗和预后情况。结果 11例IPA患者确诊2例,临床诊断9例。3例患者无特殊病史,其余8例均有基础疾病。主要症状为发热、咳嗽、咯痰、咯血、胸闷和气喘。痰涂片3例、痰培养1例、肺泡灌洗液1例、胸水1例均查见曲霉菌。1,3-β-D-葡聚糖试验(G试验)及半乳甘露聚糖试验(GM试验)阳性6例。1例支气管镜活检、1例经皮肺活检病理形态学及特殊染色结果符合曲霉菌感染。影像学表现呈多样性。11例患者均接受抗真菌药物治疗,目前仍在治疗中2例,7例随诊,失访2例。6例选用伏立康唑、3例选用伊曲康唑、2例选用卡泊芬净作为初始治疗,均有疗效。结论 IPA与基础疾病及长期使用大剂量广谱抗生素、大剂量糖皮质激素等因素密切相关,主要临床表现不典型,病情进展迅速,病死率极高。诊断有赖于病理学及病原微生学结果,但随着高分辨率CT及G试验与GM实验在临床中的开展,使早期拟诊成为可能,早期予以经验性抗真菌治疗,可明显提高IPA的治愈率。     

侵袭性肺曲霉菌病诊断方法的研究进展

侵袭性曲霉菌病(invasive aspergillosis,IA)是免疫功能低下患者最常见的侵袭性真菌感染,致死率高。由于曲霉菌孢子飘浮于空气中而易被吸入,IA中以侵袭性肺曲霉菌病(invasive pulmonary aspergillosis,IPA)最常见。然而,由于临床表现不典型,诊断金标准即病理学依据获得困难,IPA的诊断仍具挑战性,尤其是在疾病早期阶段。因此,寻找简便、快速、准确性高的诊断方法具有重要意义。本文就目前IPA的诊断进展作一综述。     

侵袭性曲霉菌病的诊断

自然界中曲霉菌超过185种，据统计大约20种曲霉菌可以引起人体机遇性感染。烟曲霉菌是最常见的一种致病曲霉菌，其次是黄曲霉、黑曲霉，而土曲霉、构巢曲霉、棒曲霉、灰绿曲霉、花斑曲霉等偶有报道引起侵袭性曲霉菌病（Invasive Aspergillosis，IA）。     

肝移植术后并发侵袭性肺曲霉菌病的原因分析及护理

目的探讨原位肝移植术后并发肺部侵袭性曲霉菌感染患者的术后护理。方法应用Meld评分、Child-Pugh评分对3例发生侵袭性肺曲霉菌病患者进行原发疾病评估。结果本组3例患者经证实发生了曲霉菌的肺部感染,均死亡。结论可将MELD评分分值高,Child-Pugh分级C级,合并糖尿病,术前存在肺部感染的患者确定为高危人群,对其进行术前评估,并给予及时、有效的综合性预防措施,可降低肝移植术后侵袭性肺曲霉菌病的发生率。     

伏立康唑治疗侵袭性肺曲霉菌病的临床观察与护理

对8例侵袭性肺曲霉菌患者实行积极的抗真菌治疗和护理,结果除1例患者死亡外,7例患者治愈或好转出院。提示治疗肺曲霉菌感染,伏立康唑为新出现的首选药,其效果可靠,不良反应少。在治疗过程中严密观察用药反应和护理尤为重要。     

侵袭性肺曲霉菌病的临床分析

近年来，随着免疫缺陷患者的增加，皮质类固醇，免疫抑制剂的应用以及抗肿瘤药物的应用，深部真菌的发病率逐年增加，曲霉菌是仅次于念珠菌的常见病原菌。侵袭性肺曲菌病（invasive pulmonary aspergillosis，IPA）为院内感染所致的深部真菌感染的一个重要原因。作者对本院自2005年1月至2007年10月收住的12例侵袭性肺曲霉菌感染患者，分析其临床特点，报道如下。     

慢性阻塞性肺疾病合并侵袭性肺曲霉菌病48例临床观察

目的 探讨慢性阻塞性肺疾病(COPD)合并侵袭性肺曲霉菌病(IPA)患者的临床特点、高危因素、治疗及转归,为COPD合并IPA的早期诊断和治疗提供依据。方法 对48例COPD合并IPA患者的临床资料进行回顾性分析。结果 48例COPD合并IPA患者主要表现为呼吸困难、肺部啰音、胸痛、咳嗽、咳痰、发热。影像学表现为肺部炎性渗出、实变、结节、空洞等。治疗有效16例,无效32例。结论 COPD合并IPA早期诊断、治疗是预后的关键。     

实验大鼠侵袭性肺曲霉病早期诊断的研究

目的 建立高效液相色谱技术（HPLC）检测（1-3）-β-D-葡聚糖的方法，并与巢式聚合酶链反应（PCR）方法进行比较，评价其在实验大鼠侵袭性肺曲霉病（IPA）早期诊断中的意义。方法 将烟曲霉孢子注入实验大鼠左肺以制作IPA模型；采集各组实验大鼠血样及脏器标本，进行巢式PCR、HPLC及培养方法的检测。结果 HPLC检测（1-3）-β-D-葡聚糖最低检测限度为1～2pg／ml，临界值为15pg／ml。模型组检测值和阳性率均较对照组高，差异有统计学意义（P〈0．01）；随感染时间延长检测值和阳性率逐渐上升，各组差异有统计学意义（P〈0．01）；感染死亡大鼠血液标本的检测值与死亡时间呈负相关关系，其相关系数r为-0．949（P〈0．01）。1周内HPLC方法敏感度（77．8％）和特异度（91．7％）均较巢式PCR高，但差异无统计学意义（P〉0．05），两者敏感度均高于血培养，差异均有统计学意义（P〈0．01）。结论HPLC检测大鼠血中（1-3）-β-D-葡聚糖浓度，在早期预测大鼠肺曲霉感染的发生、发展及预后判断方面明显优于传统的血培养，也要优于巢式PCR方法。     

Efficacy of posaconazole and amphotericin B in experimental invasive pulmonary aspergillosis in dexamethasone immunosuppressed rats

OBJECTIVES: Invasive pulmonary aspergillosis is associated with high mortality. To assess new antifungal therapy options, animal models have to be developed to assess, in an appropriate setting, the activity of new drugs. METHODS: Male albino CD rats (125-150 g) were fed with a protein-free diet and received dexamethasone thrice weekly subcutaneously during the whole experiment. After 2 weeks, an inoculum of 10(6) conidia of Aspergillus fumigatus (H11-20) was injected intratracheally. Antifungal treatment was initiated and continued for a total of 7 days. Animals were grouped in numbers of 10. One group of animals served as untreated control, whereas the others were treated with amphotericin B intraperitoneally (2 and 4 mg/kg) and posaconazole via gavage (2, 4, 10 and 20 mg/kg). Survival and log(10) cfu/g of the lungs were the endpoints. The strain H11-20 was tested for susceptibility in vitro to amphotericin B and posaconazole, respectively. Fungal burden of the lungs was expressed as log(10) cfu/g. Survival analysis was performed by the Kaplan-Meier method. Differences in fungal burden were assessed by the Mann-Whitney test. RESULTS: All untreated animals died within a week. Amphotericin B and posaconazole at 2 mg/kg demonstrated survival benefits over control (P = 0.01 and P = 0.04). Dosages of 4 mg/kg were superior to 2 mg/kg for amphotericin B (P = 0.02) and posaconazole (P < 0.05), respectively. No further survival benefits were demonstrated beyond dosages of 10 mg/kg. Rats treated with 20 mg/kg posaconazole, however, had a lower fungal burden than all the other treatment groups (P = 0.0002). CONCLUSIONS: Posaconazole and amphotericin B are effective in a dosage-dependent manner in this pulmonary aspergillosis model in immunocompromised rats.     

Novel inhalational murine model of invasive pulmonary aspergillosis

We developed a novel model of invasive aspergillosis (IA) that recapitulates human disease. Mice were immunosuppressed with cyclophosphamide and cortisone acetate and then infected in an aerosol chamber. This procedure reproducibly delivered 1 x 10(3) to 3 x 10(3) conidia to the lungs. Lethal pulmonary IA developed over 2 weeks and was prevented by amphotericin B.     

中性粒细胞减少小鼠侵袭性肺曲霉病模型的构建

目的构建中性粒细胞减少合并侵袭性肺曲霉病（IPA）的动物模型。方法40只小鼠随机分为4组：A组为模型组，环磷酰胺150mg／kg于接种前4d及接种前1d腹腔注射，接种当天给予1％戊巴比妥45mg／kg腹腔注射麻醉后，1×10^8个／mL烟曲霉孢子悬液40μL滴鼻。B组为非免疫抑制接种对照组，除以生理盐水代替环磷酰胺外，其余操作同模型组。C组为免疫抑制对照组，除以生理盐水代替烟曲霉孢子悬液滴鼻外，其余操作同模型组。D组为空白对照组，以生理盐水代替环磷酰胺腹腔注射并以生理盐水代替烟曲霉孢子滴鼻，其余操作同模型组。模型建立成功的判断标准：肺病理切片见坏死灶、肺泡内出血，PAS染色见45。角分叉的分隔菌丝或孢子；组织培养烟曲霉阳性。结果肺大体观察、病理切片H—E染色及PAS染色和肺组织培养结果表明，A组10只小鼠在接种烟曲霉3d后均发生了IPA，B、C、D组小鼠均无IPA发生。结论本法可成功构建中性粒细胞减少小鼠IPA模型。     

Efficacy of intravenous itraconazole against invasive pulmonary aspergillosis in neutropenic mice

The efficacy of itraconazole (ITZ) solubilized in hydroxypropyl-β-cyclodextrin (ITZ-IV) was examined in a murine model of invasive pulmonary aspergillosis (IPA). Immunosuppressed mice were infected by the intratracheal inoculation of Aspergillus fumigatus conidia (2 × 10⁶ conidia/mouse). Their body weight rapidly decreased and they died within 6 days after infection. Intravenous administration of various doses of ITZ-IV was started 24 h after infection and was continued once a day for 4 days. ITZ-IV at daily doses of 10, 20, or 40 mg/kg was as effective as the intraperitoneal administration of amphotericin B (AMPH) at a dosage of 1 mg/kg daily in improving survival. ITZ-IV (20 mg/kg per day), as well as AMPH (1 mg/kg per day) significantly lowered the fungal burden in the pulmonary tissues. Histological improvement was seen within 2 days after the beginning of administration of ITZ-IV (20 mg/kg per day). In mice intravenously given a single dose of ITZ-IV (20 mg/kg), the blood level and pulmonary tissue level of ITZ plus its active metabolites, mainly hydroxyitraconazole (OH-ITZ), decreased gradually after the injection, but after 4 h their concentration was still between 1.4 μg/ml (ITZ) and 1.9 μg/ml (OH-ITZ), concentrations that were approximately 10 to 20 times greater than the minimum inhibitory concentration (MIC) of ITZ for challenging the strain of A. fumigatus (0.16 μg/ml). These results support the clinical usefulness of ITZ-IV for the treatment of IPA in immunocompromised patients.     

Standardization of an experimental murine model of invasive pulmonary aspergillosis

Evaluating new therapeutic agents for invasive aspergillosis requires animal models that are reproducible among different laboratories. We therefore evaluated a murine model of aerosol infection in two independent laboratories and found a high level of both intra- and interlaboratory reproducibility of survival, fungal burden over time, and the efficacy of liposomal amphotericin B.     

侵袭性烟曲霉菌病大鼠模型的建立及分析

目的:建立侵袭性肺烟曲霉菌病大鼠模型,并对模型研究价值进行相关分析,为侵袭性肺曲霉菌病的实验研究提供参考.方法:腹腔注射环磷酰胺,动态观察大鼠血常规变化,建立免疫抑制大鼠模型.自制气管插管接种烟曲霉菌孢子悬液引起侵袭性肺烟曲霉菌病,分别于接种后1、3、5、7 d处死大鼠后取肺组织进行病理切片观察及组织匀浆培养.结果:分次不同剂量环磷酰胺腹腔注射后大鼠表现为精神萎靡、体重减轻,血常规示白细胞及分类细胞数量明显减少,中性粒细胞、淋巴细胞不能分类.病理切片可见免疫抑制大鼠感染烟曲霉菌后随着时间延长肺组织病变逐渐加重.肺组织出血、纤维素渗出;烟曲霉菌孢子聚集并长出菌丝,引起肺泡结构破坏、弥漫性肺水肿.结论:成功建立了免疫抑制大鼠侵袭性肺烟曲霉菌病动物模型,为研究免疫抑制宿主侵袭性肺曲霉菌病的发病机制、病理生理、药物疗效提供参照.     

Efficacy of voriconazole against invasive pulmonary aspergillosis in a guinea-pig model

We compared the efficacies of amphotericin B and voriconazole against invasive pulmonary aspergillosis in a guinea-pig model. A susceptible isolate of Aspergillus fumigatus was used to produce the infection. Voriconazole-treated animals had significantly better survival and decreased fungal burden in the lungs as compared with controls. Although no statistical difference was seen between the efficacies of voriconazole and amphotericin B, a trend favouring voriconazole was noted. Thus, voriconazole, with its cidal activity, may be an attractive alternative to potentially toxic amphotericin B in the treatment of invasive pulmonary aspergillosis.