目的:对引起中枢神经系统感染的一株罕见粘液型溶血葡萄球菌(S.haemolyticus)表型及基因组特征进行分析.方法:收集一株导致中枢神经系统感染粘液型溶血葡萄球菌,通过生物膜形成实验、药敏实验分析生物膜形成和耐药表型,全基因组测序分析其耐药等基因特征.结果:本例溶血葡萄球菌呈粘液型,可见拉丝现象,属于苯唑西林耐药凝固酶阴性葡萄球菌(MRCoNS),同时对红霉素、克林霉素、左氧氟沙星耐药,对万古霉素、利奈唑胺敏感,比普通非粘液型溶血葡萄球菌更易形成生物膜,MLST显示该菌为ST1型,携带mecA、aph(3')-Ⅲa、aac(6')-Ie、aph(2")-Ia等8种耐药基因,mecA基因由类似Tn6191的转座子所携带.结论:呈粘液型菌落特征的溶血葡萄球菌在临床上极为罕见,该变异体比通常的非粘液型菌落更易形成生物膜且呈现出多重耐药现象.临床上应关注此类葡萄球菌的粘液变异体,这有助于其早期识别和感染防控.

The first patient complained of red eye, photophobia, and difficulty in eye-opening of the right eye for 5 months. The second patient complained of grinding pain and difficulty in eye-opening of the left eye for 1 week. The third patient complained of repeated red eye and blurred vision in the left eye for half a year. After detailed examination, a blue suture thread approximately 2 mm in length with a knot was found in the mucosal fold of the first patient, a bee sting was embedded in the middle of the meibomian gland duct in the upper eyelid of the second patient, a glassy foreign body was found in the chamber angle at the 6 o'clock position of the third patient. The corneal and ocular surface injury gradually healed with no recurrence after the timely removal of the occult foreign bodies.

目的:了解山东第一医科大学第一附属医院血流感染肺炎克雷伯菌的临床分布及耐药特征,为临床抗感染治疗提供理论依据.方法:收集山东第一医科大学第一附属医院2017年～2019年引发血流感染的肺炎克雷伯菌菌株,使用MALDI-TOF1MS进行菌种鉴定,应用纸片法进行药敏试验.结果:共分离到196株肺炎克雷伯菌,ICU病区及60岁以上的老年患者是主要的感染人群,男性患者人数要高于女性患者.除氨苄西林、 头孢唑林及头孢噻肟外,其余抗生素的耐药率均在50％以下.但不同抗生素在不同年份表现出不同的变化规律.结论:ICU病区、 老年患者及男性患者应该成为血流感染肺炎克雷伯菌的重点关注人群.鉴于本院血流感染肺炎克雷伯菌中多重耐药菌尤其是CRE菌株具有较高的分离率且不同抗生素的耐药率呈现不同变化趋势,临床上需要根据药敏结果合理用药.

Objective::To analyze the pathogenesis, clinical manifestations and prognosis of candida keratitis.Methods::In this retrospective case series study, a total of 29 patients (29 eyes) who were diagnosed with candida keratitis from January 2013 to December 2019 were recruited. There were 18 males and 11 females, all with monocular disease. One patient was a 7-month-old girl, and the other 28 patients were between 29 and 91(59.0±13.5) years old. The onset time ranged from 2 to 90 days, with a median of 12 days. The incidence trend, the condition before infection, clinical manifestations, image characteristics of confocal microscopy, treatment and curative effect were analyzed.Results::A total of 29 cases were positive for corneal candida culture (10 cases of candida albicans, 6 cases of candida parapsilosis, and 13 other cases), accounting for 2% of all patients with fungal keratitis during the same period. The most common the condition before infection, were ocular surgery in 26 patients (90%), followed by a history of topical glucocorticoid use in 24 patients (83%), systemic disease in 9 patients (31%), and ocular trauma in 2 patients (7%). Twenty-five patients (86%) had two or more risk factors. The conjunctival congestion of patients with candida keratitis was generally mild, without the typical signs of pseudopodia, hyphae moss, or immune ring structure. Four patients with early keratopathy presented with single or multiple isolated, rounded lesions, which were cured by corneal lesion resection combined with drug therapy. After the lesion progression, the lesions showed various manifestations. The most common was scattered multi-focal infiltration (10 cases, 35%), and the final 23 cases (80%) required surgical treatment.Conclusions::A history of ocular surgery and topical glucocorticoid use are common conditions. The early stage of the disease is characterized by isolated, rounded lesions and there is good prognosis if patients are treated with corneal lesion resection combined with drug therapy. Most patients at the late stage require keratoplasty.

Objective:To analyze the molecular characteristics and antibiotic susceptibility of two strains of Nocardia farcinica isolated from patients with joint infection using whole genome sequencing. Methods:Two strains of Nocardia farcinica causing knee-joint infections in two elderly patients were collected in January 2020. Whole genome sequencing was used to determine the nocardia species. Drug sensitivity test was performed using the micro-broth dilution and E-test method according to CLSI M24 guideline. ABRicate was used to analyze drug resistance and virulence genes. Snippy and other bioinformatic tools were used for genomic comparison, and to construct SNP homologous tree. Results:The clinical isolates in this study were both Nocardia farcinica. Antimicrobial susceptibility test showed the isolates were resistant to ceftriaxone, cefepime, cefotaxime and trimethoprim/sulfamethoxazole (TMP/SMX). Imipenem, linezolid and amoxicillin-clavulanic acid showed good activity. Four antibiotic resistance genes including class A β-lactamase gene far-1, RNA polymerase binding protein gene RbpA, multi-drug resistance efflux pump transcription activator gene MtrA and regulatory transcription factor gene vanR-O were identified in the Nocardia farcinica genomes, which conferred resistance to beta-lactams, rifampicin, macrolides and vancomycin respectively. No acquired TMP/SMX resistance genes were identified. There are multiple missense mutations in the dihydrofolate reductase family genes. Four virulence genes of icl, mbtH, phoP,and relAthat are homologous to Mycobacterium tuberculosis were found. SNP homologous tree analysis showed the two Nocardia strains were closely related, and there were only ten SNP sites, six compound substitutions and one deletion mutation between them. Conclusions:Whole genome sequencing technology is helpful to explore the molecular characteristics and resistance mechanisms of Nocardia species. Nocardia farcinica has a trend of spreading in China. Resistance to TMP/SMX is worthy of attention. The mutation of genes involved in the metabolic pathway of dihydrofolate might be one of multiple TMP/SMX resistance mechanisms.

目的:了解2019年山东千佛山医院临床细菌分离株的分布及耐药性,为临床合理使用抗生素提供依据.方法:通过Bruker MALDI-TOF、Walkaway 96 plus或VITEK-2 Compact自动化细菌鉴定仪进行细菌鉴定,纸片扩散法 、Walkaway 96 plus或VITEK-2 Compact自动化药敏系统进行药物敏感性测定.药敏按照2019年CLSI M100中规定的折点.使用WHONET 5.6软件分析数据.结果:2019年1月～12月共检出7634株临床分离株,其中革兰氏阴性菌和革兰阳性菌分别占70.5％和27.4％.前五种病原体分别是大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌、金黄色葡萄球菌以及鲍曼不动杆菌.甲氧西林耐药金黄色葡萄球菌(MRSA)和凝固酶阴性葡萄球菌(MRCNS)占比分别为22.1％和31％.没有检测到对万古霉素,替考拉宁或利奈唑胺耐药的葡萄球菌.0.7％的屎肠球菌菌株对万古霉素耐药.分离出239株耐碳青霉烯的肠杆菌科细菌,占所有肠杆菌科分离株的7.6％.不动杆菌属对亚胺培南和美罗培南耐药率分别为72.2％和71.9％.铜绿假单胞菌对亚胺培南和美罗培南的耐药率分别为17.3％和14.3％.结论:我院的多重耐药细菌较前两年呈下降趋势,但形势仍较严峻,医院应做好抗菌药物临床使用管理,加强感染控制措施的落实,同时保持良好的细菌耐药性监测.

Objective::To use in vivo laser confocal microscopy (IVCM) to observe the in vitro culture and living characteristics of candida spores and pseudohyphaes, and to provide abasis for the early diagnosis of candida keratitis.Methods::The data of 26 patients (26 eyes) who were diagnosed with candida keratitis in the Eye Hospital of Shandong First Medical University from January 2016 to December 2019 was collected. At the same time, candida albicans and candida parapsilosis were taken for laboratory fungal culture, and the typical morphology of candida spores and pseudohyphae in vitro and in vivo were observed by IVCM.Results::Candidaspores in vitro and in vivo presented uniformly in the shape of round spots with a diameter of 2-5 μm and a clear boundary. Pseudohyphae have a highly reflective linear structure. The reflection of spores was significantly enhanced in vivo. Eighteen eyes showed a scattered arrangement of spot-like, strongly reflective structure, and 11 eyes showed a dense fog structure. The morphology of pseudomycelia varied greatly. The mycelia in 2 eyes were slender, the spore structure was present in 5 eyes, and branches were visible. In 1 eye, an irregular thin and long rod shape with high contrast was observed, and in another eye, an irregular thin and long rod shape and spore structure could be observed simultaneously. Candida albicans pseudohyphae were relatively slender, with a length of 20-150 μm and a diameter of 2-10 μm. Pseudohyphae of candida parapsilosis were short and thick, with a length of about 30-100 μm and a diameter of 2-5 μm. Part of the branch spore structure was highlighted.Conclusions::Candida keratitis showstypical spore and pseudohyphae structures with IVCM. The morphology of candida keratitis spores in vitro and in vivo are consistent with observations under IVCM, but the morphology of pseudohyphaes are relatively different. IVCM can be used for the early diagnosis of candida keratitis.

目的:了解2017年山东千佛山医院临床细菌分离株的分布及耐药性,为临床合理使用抗生素提供依据.方法:通过Bruker MALDI-TOF、Walkaway 96 plus或VITEK-2 Compact自动化细菌鉴定仪进行细菌鉴定,纸片扩散法、Walkaway 96 plus或VITEK-2 Compact自动化药敏系统进行药物敏感性测定.药敏按照2016年CLSIM100中规定的折点,使用WHONET 5.6软件分析数据.结果:2017年1月～12月共检出6511株临床分离株,其中革兰氏阴性菌和革兰阳性球菌分别占74.5％和25.5％.前五种病原体分别是大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌、鲍曼不动杆菌和金黄色葡萄球菌.对甲氧西林耐药的金黄色葡萄球菌(MRSA)和凝固酶阴性葡萄球菌(MRCNS)分离率分别为25.5％和77.6％.没有检测到对万古霉素,替考拉宁或利奈唑胺耐药的葡萄球菌.4.3％的屎肠球菌菌株对万古霉素耐药.分离出239株耐碳青霉烯的肠杆菌科细菌,占所有肠杆菌科分离株的8.5％.不动杆菌属对亚胺培南和美罗培南耐药率分别为81.7％和83％.铜绿假单胞菌对亚胺培南和美罗培南的耐药率分别为34.2％和32.4％.结论:我院的多重耐药细菌形势严峻,应加强医院感染控制和抗菌药物临床使用管理,保持良好的细菌耐药性监测.

肠杆菌科细菌是临床上较常见致病菌, 通过产生超广谱β-内酰胺酶(ESBLs) 及AmpC酶等达到多重耐药的效果, 目前亚胺培南、 厄他培南等碳青霉烯类抗生素属于治疗该类细菌较为有效的抗菌药物, 也被称为最后一道防线[1]. 耐碳青霉烯类肠杆菌科细菌(carbapenem-resistant Enterobacteriaceae,CRE)的出现突破了最后防线, 延长了患者住院时间, 提高了死亡风险, 严重威胁着人类的生命健康[2-3]. 为指导临床进行防范及合理化用药, 我们回顾性分析了2016年1月～2017年6月期间我院所有耐碳青霉烯类肠杆菌科细菌及其耐药情况. 具体情况如下.