碳青霉烯类耐药肺炎克雷伯菌基因型检测研究

目的 分析碳青霉烯类耐药肺炎克雷伯菌(CRKP)不同基因型耐药性及临床特征差异，为临床预防及治疗CRKP感染提供参考。方法 回顾性分析我院2018年9月—2021年8月收治的56例分离出CRKP菌株患者的临床资料，检测其耐药基因，并分析不同基因型CRKP的耐药性和临床特征。结果 56株CRKP中检出47株KPC型耐药基因(83.93%)、9株NDM-1型耐药基因(16.07%),未检出IMP、VIM以及OXA-48耐药基因；耐药性分析结果发现，KPC和NMD-1型耐药基因对氨曲南耐药性差异有统计学意义(P<0.05);KPC和NMD-1型耐药基因对应患者年龄、性别、收治科室、标本来源分布情况、合并基础疾病、抗菌药物使用时间、3种及以上抗菌药物联用史、住院时间以及侵入性操作比较差异均无统计学意义(P>0.05)。结论 CRKP基因型主要以KPC型最常见，不同基因型CRKP临床特征无明显差异，不同基因型CRKP对常用抗菌药物耐药性存在异质性，临床抗感染治疗需结合基因检测及耐药性分析制定合理方案。     

医院内鲍曼不动杆菌感染分布特征及其耐药机制

目的 了解天津市某医院临床分离鲍曼不动杆菌(Ab)的分布特征及耐药性,并探讨Ⅰ类整合子与重症监护病房(ICU)来源Ab耐药的关系.方法 回顾性分析2010年7月～2011年6月该院住院患者的Ab标本来源、科室分布及耐药性,PCR结合基因序列分析检测同期该院ICU来源的57株Ab的Ⅰ类整合子.结果 共分离616株Ab,来自ICU 412株(66.9％),来自呼吸道标本512株(83.1％).Ab对多粘菌素B、头孢哌酮/舒巴坦和左氧氟沙星的耐药率分别为0、13.8％、45.8％,对其他抗菌药物耐药率均在55.0％以上,多重耐药Ab达76.6％.57株来自ICU的Ab中,Ⅰ类整合酶阳性43株;39株可变区阳性菌基因序列分析显示,携带aacA4-catB8-aadA1、aacCl-orfA-orfB-aadA1两种形式的耐药基因盒.结论 该院Ab耐药情况严重,尤其是ICU来源的Ab更应引起足够重视;Ⅰ类整合子与ICU分离Ab的耐药性有关.     

2006～2009年103株嗜麦芽窄食单胞菌医院内感染危险因素及药敏分析

目的 了解嗜麦芽窄食单胞菌的医院感染现状及耐药状况,为临床合理使用抗菌药物提供参考.方法 分析我院2006年1月～2009年12月间临床患者分离的103株嗜麦芽窄食单胞菌的感染情况及耐药状况.结果 103株嗜麦芽窄食单胞菌多见于呼吸道感染(81.55％).科室分布中,ICU占40.79％,呼吸内科占17.48％.嗜麦芽窄食单胞菌对多种抗菌药物耐药,对复方新诺明耐药率最低(28.2％),左氧氟沙星次之(34.0％).结论 嗜麦芽窄食单胞菌对常用药物呈现多重耐药,要合理使用抗菌药物,控制医院感染的发生.     

我院2009～2010年鲍曼不动杆菌的临床分布及耐药性分析

目的了解鲍曼不动杆菌在我院各临床科室的分布情况以及耐药性,为临床合理使用抗菌药物提供依据。方法对我院2009年1月～2010年12月住院患者标本进行鲍曼不动杆菌分离、鉴定及耐药性分析。结果①共分离出非重复鲍曼不动杆菌243株,主要分离自痰标本,其次是血和创面分泌物;病区分布依次是ICU、呼吸内科和神经外科。②鲍曼不动杆菌对阿米卡星耐药率最低,为14.5%;对其他12种抗菌药物的耐药率均>70.0%;已出现大量多重耐药菌株及少数泛耐药菌株。结论我院鲍曼不动杆菌感染主要见于ICU、呼吸内科病房等,且对抗菌药物已产生明显多药耐药性;应警惕并重视该菌感染及耐药性监测,预防医院感染的发生。     

某综合医院严格实施多重耐药菌管理综合干预措施1年效果观察

目的观察预防控制措施对降低多重耐药菌医院感染率的效果,为制定各项预防控制措施提供依据。方法我院于2014年12月开始严格实施多重耐药菌管理综合干预措施。选取2014年1月1日—2015年12月31日期间的住院患者,分别统计住院患者的抗菌药物使用率、抗菌药物使用强度及微生物标本送检率,对检出多重耐药菌感染或定植的住院患者,分别统计病房预防控制措施落实情况、多重耐药菌检出率及多重耐药菌医院感染率。结果与2014年数据相比,2015年我院抗菌药物使用率下降,治疗性使用抗菌药物的微生物标本的送检率均有上升;耐碳青霉烯类铜绿假单胞菌(carbapenem-resistant Pseudomonas aeruginosa,CRPA)、耐碳青霉烯类肺炎克雷伯菌(carbapenem-resistant Klebsiella pneumoniae,CRKP)的检出率明显下降,产超广谱β-内酰胺酶(extended spectrumβ-lactamases,ESBLs)肺炎克雷伯菌的检出率有所上升;隔离标识放置率、手卫生设施落实率、患者生活垃圾正确处置率均有上升;CRKP和产ESBLs肺炎克雷伯菌医院感染发生率下降,差异有统计学意义(P均0.05);耐碳青霉烯类鲍曼不动杆菌、CRPA医院感染发生率下降,耐甲氧西林金黄色葡萄球菌、产ESBLs大肠埃希菌医院感染发生率上升,但差异无统计学意义(P均0.05)。结论合理使用抗菌药物,降低抗菌药物使用率,落实病房预防控制措施可以有效的降低多重耐药菌医院感染率。     

136株住院患者感染铜绿假单胞菌的耐药状况及机制研究

目的分析铜绿假单胞菌的耐药状况及耐药基因携带情况,为探究菌株耐药机制、控制耐药性发展提供科学指导。方法收集本院各科室患者的送检标本,采用全自动细菌鉴定仪鉴定并保存待用。采用K-B法进行药敏性实验,并根据2015版CLSI标准判定菌株耐药程度。然后,采用PCR扩增法检测铜绿假单胞菌的耐药基因。结果共分离136株铜绿假单胞菌,其中ICU 44株,呼吸内科29株,神经外科19株,泌尿外科12株,普外科10株,消化内科7株,其他科室15株,构成比分别为32.35%、21.32%、13.97%、8.82%、7.35%、5.15%、11.03%。ICU病房、呼吸内科、神经内科是铜绿假单胞菌的主要来源。痰液标本来源102株,伤口分泌物10株,脓液9株,尿液5株,血液标本3株,其他标本来源7株,菌株构成比分别为75.00%、7.35%、6.62%、3.68%、2.21%和5.15%。分离菌株对哌拉西林、头孢噻肟、头孢他啶、庆大霉素、氨曲南、头孢吡肟、亚胺培南、妥布霉素、阿米卡星的耐药率分别为100.00%、53.68%、33.09%、27.94%、25.74%、21.32%、19.12%、16.91%和13.24%。铜绿假单胞菌分离株各耐药基因检出率:TEM基因67.65%,VEB基因21.32%,IMP基因11.03%,VIM基因13.24%,aac(6′)-Ib基因16.91%,aac(6′)-II基因24.26%,ant(2′)-I基因25.74%,oprD2基因13.97%。结论铜绿假单胞菌主要分离自医院ICU及患者的痰液标本,临床抗感染治疗时应优先考虑阿米卡星类抗菌药物。TEM基因耐药基因可能与菌株对β-内酰胺类抗菌药物的耐药性有关。     

下呼吸道铜绿假单胞菌感染的耐药性变迁分析

目的了解本院ICU下呼吸道感染铜绿假单胞菌(PAE)的耐药性变化趋势。方法分析2010年1月~2013年12月ICU下呼吸道痰标本分离到的276株PAE药敏结果。结果亚胺培南、美洛培南耐药率呈上升趋势;庆大霉素、阿米卡星耐药率呈下降趋势;其余11种耐药率变化趋势没有显著差异。2012年比2011年的多重耐药株(MDRPAE)及泛耐药株(PDRPAE)检出率呈升高趋势。结论部分抗菌药物对PAE耐药性呈上升趋势,部分年份MDRPAE及PDRPAE有升高的趋势,应重视对PAE的耐药性监测,并采取措施减少耐药菌株的产生。     

医院感染鲍氏不动杆菌的临床分布及耐药性分析

目的:了解2008-01-20120-12临床分离的221株鲍氏不动杆菌(ABA)的标本来源、感染科室分布及耐药状况,为临床预防和治疗感染性疾病提供参考依据。方法:采用临床微生物学检验的常规方法对感染性标本进行细菌培养、鉴定和药敏试验,调查221株ABA的标本来源和感染科室,统计ABA对12种常用抗菌药物的耐药率。结果:221株ABA中有182株来自痰液,占82.4%;其次是脓液和手术切口引流物、血液等。医院感染ABA主要分布在重症监护病房(ICU),占51.6%;其次是呼吸内科24.4%和神经外科19.5%。药敏结果显示:ABA对临床常用抗生素的耐药性逐年增长,ABA耐药率最低的抗生素是头孢哌酮/舒巴坦,其次是亚胺培南、美洛培南。对青霉素类、氨基糖苷类、氟喹诺酮类、氨曲南、哌拉西林/他唑巴坦等呈高度耐药(90%),且表现出多药耐药性。结论:ABA医院获得性感染率逐年增加,并对多种抗菌药物耐药,且多药耐药性明显,应引起临床尤其是ICU病房和呼吸内科高度重视,应严格做好各种侵入性操作的无菌技术,合理使用抗生素,以降低医院感染率、提高治愈率,延缓细菌耐药性的产生。     

250株鲍曼不动杆菌临床分离株耐药性分析

目的了解鲍曼不动杆菌临床分布及其耐药性,为防治院内感染提供依据。方法对临床分离的250株鲍曼不动杆菌进行回顾性分析,计算对常用的15种抗生素的耐药率。结果 250株鲍曼不动杆菌中,2008年74株(占29.6%),2009年105株(占42.0%),2010年71株(占28.4%)。科室分布以ICU(77株)、呼吸内科(59株)和神经外科(40株)为主;标本分布以痰标本为主(216株),其次为脑脊液标本(10株);药敏试验除对头孢哌酮/舒巴坦耐药率较低外,对其余14种抗生素耐药率较高,表现为多重耐药,且呈逐年升高趋势;与ICU、呼吸内科、神经内科和心胸外科分离菌株相比,老年病区及其他散在病区分离菌株的耐药率相对较低。结论鲍曼不动杆菌多重耐药现象严重,应加强耐药性的监测,了解耐药变迁情况,以指导临床合理用药。     

基层医院呼吸内科肺炎克雷伯菌感染现况和耐药性分析

目的：了解基层医院呼吸内科肺炎克雷伯菌的耐药现况和评价《抗菌药物临床应用专项整治活动方案》的实施效果,为临床医师合理应用抗菌药物提供科学依据。方法用 VITEK2微生物分析系统对本院2012年1月至2013年12月呼吸内科标本中分离的肺炎克雷伯菌进行药敏分析,比较分析《抗菌药物临床应用专项整治活动方案》干预前(2012年)和干预后(2013年)呼吸内科肺炎克雷伯菌的耐药性。结果2012年(干预前)和2013年(干预后)从本院呼吸内科标本分离的肺炎克雷伯菌分别为73株和58株,其主要来源于呼吸系统感染,分别占91.8%和82.8%;该项方案干预后,产超广谱β-内酰胺酶肺炎克雷伯菌检出率由27.4%降为20.7%,其对常用β-内酰胺类药物的耐药率低于干预前,尤其是氨曲南、氨苄西林/舒巴坦、头孢唑啉、头孢呋辛、头孢曲松、头孢他啶和头孢吡肟的耐药率显著降低(P <0.05)。结论该项方案干预后,呼吸内科标本分离的肺炎克雷伯菌对常用13种抗菌药物耐药率低于干预前,其中有10种抗菌药物的耐药率显著降低。     

住院患者849例抗菌药物应用调查

目的了解医院抗菌药物临床应用的现状及合理性，找出相应的管理措施。方法根据设计的调查表，按照统一方法和标准对12个科室849例外科出院患者回顾性调查。结果抗菌药物总使用率89．40％，其中预防性应用占总使用率的70．00％，青霉素、头孢类药物使用率最高，其次为喹诺酮类，一联用药474例，二联用药236例，三联及以上49例，应用疗程≤7d的529例。结论应加大对抗菌药物使用的管理力度，加强对医务人员用药知识的培训，达到抗菌药物合理应用。     

Therapeutic potential of Takeda‐G‐protein‐receptor‐5 (TGR5) agonists. Hope or hype?

The gastrointestinal tract regulates glucose and energy metabolism, and there is increasing recognition that bile acids function as key signalling molecules in these processes. For example, bile acid changes that occur after bariatric surgery have been implicated in the effects on satiety, lipid and cholesterol regulation, glucose and energy metabolism, and the gut microbiome. In recent years, Takeda‐G‐protein‐receptor‐5 (TGR5), a bile acid receptor found in widely dispersed tissues, has been the target of significant drug discovery efforts in the hope of identifying effective treatments for metabolic diseases including type 2 diabetes, obesity, atherosclerosis, fatty liver disease and cancer. Although the benefits of targeting the TGR5 receptor are potentially great, drug development work to date has identified risks that include histopathological changes, tumorigenesis, gender differences, and questions about the translation of animal data to humans. The present article reviews the noteworthy challenges that must be addressed along the path of development of a safe and effective TGR5 agonist therapy.     

Changes in medication use from age 26 to 32 in a representative birth cohort

Background: To date, longitudinal studies of medications have been confined to older adults or clinical samples, with no data from prospective studies of younger adults. The aim of the study was to examine changes in medication usage between ages 26 and 32 in a prospective study of a representative birth cohort. Methods: Medication use during the previous 2 weeks was investigated among 960 individuals at ages 26 and 32. Results: Nearly two‐thirds took at least one medication at each age, with medication prevalence higher among women than among men. Three‐quarters of those taking at least one at age 26 were doing so at 32. Over‐the‐counter medication prevalence increased from 35 to 43% between 26 and 32 years of age. Although the prevalence of prescribed medications decreased (from just under half to just over one‐third, and from two‐thirds to below half among women), there was no significant difference between the ages once hormonal contraceptives were accounted for. By 32, reduced usage of hormonal contraceptives was apparent, with one‐third of age‐26 users still taking these at 32. Other categories showing major changes were analgesics (increased), anti‐asthma drugs (decreased), antidepressants (increased) and antiulcer drugs (increased). At 32, 82% of those taking analgesics, 85% of those taking nutrient supplements, 71% of those taking antihistamines and 33% of those taking antiulcer drugs had self‐prescribed them. Conclusion: A considerable proportion of the sample used medications by age 32, and there was considerable change between 26 and 32. The changes are likely to have been due to a mix of ageing and period effects.     

心血管内局部定位药物缓释体系的实验研究

本研究探讨了载药纳米微球(NP)作为血管内靶向定位药物控释载体的可行性,通过最佳配方和导入条件的筛选为临床研究提供了基础.用聚乳酸-乙醇酸共聚物(PLGA)制备了包载抗细胞增生药物细胞松驰素B的生物降解性纳米微球.用狗为实验动物研究了正常血液循环条件下,纳米微球在血管内的吸收和定位的可能性和最佳条件.结果表明载药纳米微球可穿透结缔组织并被靶部位的血管壁吸收,可以用介入方法将NP导入血管内病灶部位,并使其在血管局部组织内缓慢释放药物,从而维持长期局部有效药物浓度,可达到有效地治疗心血管再狭窄及其他血管疾病的目的.     

Thrombocytopenic purpura in HIV-seronegative users of intravenous cocaine

Severe thrombocytopenic purpura occurred in seven patients who had histories of recent or current intravenous cocaine use. All of the patients denied heroin use and all were seronegative for antibodies to the human immunodeficiency virus (HIV). The seven patients had normal or increased numbers of megakaryocytes in their bone marrows. While receiving corticosteroid therapy their platelet counts rose to 10(11)/L over periods ranging from 5 to 11 days. In view of the widespread use of cocaine, severe thrombocytopenia in the HIV-seronegative patient should suggest the possibility of current or recent intravenous use of cocaine.     

Preemployment drug screening in a large metropolitan medical center

To assess the prevalence of illicit drug use among job applicants, a large metropolitan medical center conducted preemployment drug screening of all applicants during January 1988. Urine samples from 172 preinformed applicants were screened using Enzyme Multiplied Immunoassay Technique (Emit® d.a.u.?) followed by confirmatory gas chromatography/mass spectrophotometry. 4.1% of tests were positive for marijuana and/or cocaine and none was positive for beroin. Positive findings increased with decreasing socioeconomic status. The findings suggest that applicants for jobs in large medical centers in metropolitan areas are no different from those in other sectors of the economy with respect to illicit drug use.     

Development of covalent inhibitors that can overcome resistance to first-generation FGFR kinase inhibitors

The human FGF receptors (FGFRs) play critical roles in various human cancers, and several FGFR inhibitors are currently under clinical investigation. Resistance usually results from selection for mutant kinases that are impervious to the action of the drug or from up-regulation of compensatory signaling pathways. Preclinical studies have demonstrated that resistance to FGFR inhibitors can be acquired through mutations in the FGFR gatekeeper residue, as clinically observed for FGFR4 in embryonal rhabdomyosarcoma and neuroendocrine breast carcinomas. Here we report on the use of a structure-based drug design to develop two selective, next-generation covalent FGFR inhibitors, the FGFR irreversible inhibitors 2 (FIIN-2) and 3 (FIIN-3). To our knowledge, FIIN-2 and FIIN-3 are the first inhibitors that can potently inhibit the proliferation of cells dependent upon the gatekeeper mutants of FGFR1 or FGFR2, which confer resistance to first-generation clinical FGFR inhibitors such as NVP-BGJ398 and AZD4547. Because of the conformational flexibility of the reactive acrylamide substituent, FIIN-3 has the unprecedented ability to inhibit both the EGF receptor (EGFR) and FGFR covalently by targeting two distinct cysteine residues. We report the cocrystal structure of FGFR4 with FIIN-2, which unexpectedly exhibits a “DFG-out” covalent binding mode. The structural basis for dual FGFR and EGFR targeting by FIIN3 also is illustrated by crystal structures of FIIN-3 bound with FGFR4 V550L and EGFR L858R. These results have important implications for the design of covalent FGFR inhibitors that can overcome clinical resistance and provide the first example, to our knowledge, of a kinase inhibitor that covalently targets cysteines located in different positions within the ATP-binding pocket.Receptor tyrosine kinases (RTKs) serve as critical sensors of extracellular cues that activate a myriad of intracellular signaling pathways to regulate cell state. There are 58 receptor tyrosine kinases in the human genome, and many have been demonstrated to be constitutively activated through amplification or mutation in particular cancers. The signals emanating from these RTKs, such as epidermal growth factor receptor (EGFR), FGF receptor (FGFR), platelet-derived growth factor receptor (PDGFR), protein kinase Kit (KIT), and protein kinase c-Met (MET), have been pharmacologically proven to be essential to the survival of cancers expressing mutant forms of these proteins. However, rapid resistance to monotherapy with first-generation RTK inhibitors has been universally observed. Resistance typically arises from the emergence of cancer cells expressing mutant forms of RTKs that are impervious to the action of first-generation drugs or from the activation of by-pass signaling mechanisms. Resistance can be overcome by developing new inhibitors that target the mutant RTK directly or target bypass signaling mechanisms. Indeed this approach has been deployed successfully in the case of resistance to first-generation inhibitors of EGFR in nonsmall cell lung cancer (NSCLC) and of Abelson tyrosine-protein kinase (ABL) in chronic myelogenous leukemia (CML) (1–4).Human FGFRs are a family of four RTKs (FGFR1–4) which are sensors of a diverse family of 18 FGF ligands. FGFRs are key regulators of fibrogenesis, embryogenesis, angiogenesis, metabolism, and many other processes of proliferation and differentiation (5, 6). The fundamental importance of FGFR to development is well proven by gain-of-function mutations that result in dwarfism in model organisms and in humans (7–10). Deregulation of FGFR signaling through mutation, chromosomal translocation, and gene amplification or overexpression has been documented abundantly in numerous cancers (11). Activation of FGFR-dependent signaling pathways can stimulate tumor initiation, progression, and resistance to therapy. Translocation events implicating the FGFR1 gene and various fusions of FGFR1 are found in myeloproliferative syndromes (12); chromosomal translocations of FGFR1 or FGFR3 and the transforming acidic coiled-coil genes (TACC1 or TACC3) are oncogenic in glioblastoma multiforme, bladder cancer, head and neck cancer, and lung cancer (13–16); oncogenic mutations of FGFR2 and FGFR3 are observed in lung squamous cell carcinoma; FGFR2 N549K is observed in 25% of endometrial cancers; FGFR3 t(4;14) alterations are reported in 15–20% of multiple myeloma (17–19); FGFR4 Y367C mutation in the transmembrane domain drives constitutive activation and enhanced tumorigenic phenotypes in a breast carcinoma cell line (20–22); and K535 and E550 mutants are reported to activate FGFR4 in rhabdomyosarcoma (23). FGFR amplification is reported in various cancers (24, 25): FGFR1 is amplified in colorectal, lung, and renal cell cancers (26, 27); FGFR2 is amplified in gastric cancer and colorectal cancer (28, 29); FGFR3 is commonly amplified in bladder cancer and also is reported for cervical, oral, and hematological cancers (30–32); and FGFR4 is amplified in hepatocellular carcinoma, gastric cancer, pancreatic cancer, and ovarian cancer (33–37). FGFR also is involved in autocrine activation of STAT3 as a positive feedback in many drug-treated cancer cells which are driven by diverse oncogenes such as EGFR, ALK, MET, and KRAS (38).Currently known inhibitors of kinases can target a variety of conformational states and binding pockets and can be either reversible or covalent. Several potent and selective ATP-competitive, small-molecule FGFR inhibitors have been reported, with BGJ398 and AZD4547 being the clinically most advanced compounds (Fig. 1A) (39–42). We previously reported the first (to our knowledge) covalent FGFR irreversible inhibitor (FIIN-1), which targets a cysteine residue conserved in all four FGFR kinases and which inhibits the proliferation of Ba/F3 cells engineered to be dependent on FGFR1, FGFR2, or FGFR3 with EC₅₀s in the 10-nM range, a potency comparable to that exhibited by BGJ398 and AZD4547 (43). All FGFR kinases have a valine at the gatekeeper position, in contrast to ABL, EGFR, KIT, and PDGFR, which all possess a threonine gatekeeper in which resistance can be conferred by mutation of the threonine to a larger hydrophobic valine, isoleucine, or methionine residue in response to first-generation inhibitors of these kinases (44–46). The FGFR V561M mutation was reported to induce strong resistance to PD173074 and FIIN-1 (43, 47); later the gatekeeper mutant FGFR3 V555M emerged as a mechanism of resistance to AZ8010 in KMS-11 myeloma cells and also was demonstrated to confer resistance to other FGFR inhibitors, including PD173074 and AZD4547 (48). The FGFR2 V564I gatekeeper mutant was isolated as a resistant clone in a FGFR2 Ba/F3 screen of dovitinib and also was reported to confer resistance to the multitargeted drug ponatinib (19). In humans the FGFR4 V550L gatekeeper mutation was detected in 9% (4/43) of embryonal rhabdomyosarcoma tumors (49), and the FGFR4 V550M mutation was detected in 13% (2/15) of neuroendocrine breast carcinomas (50). To overcome gatekeeper mutations found in primary FGFR-driven cancers and those that likely will arise in FGFR inhibitor-treated tumors in the future, we developed next-generation covalent FGFR inhibitors. Here we describe the identification and characterization of the covalent FGFR inhibitors FIIN-2 and FIIN-3, which to our knowledge are the first FGFR inhibitors that are capable of potently inhibiting the gatekeeper mutants of FGFRs. We also demonstrate that FIIN-3 is capable of covalently inhibiting both FGFR and EGFR by using distinct binding modes to target different cysteine residues.Open in a separate windowFig. 1.(A) Chemical structures of clinical-stage FGFR inhibitors. (B) Evolution of FIIN-2 and FIIN-3 from FIIN-1. Structures of the reversible counterparts FRIN-2 and FRIN-3 are shown also.