Lower respiratory tract infection is one of the major causes of disease and death worldwide. Streptococcus pneumoniae, Mycoplasma pneumoniae, and Haemophilus influenzae are important pathogens responsible for lower respiratory tract infection. Here, we established a multiplex droplet digital polymerase chain reaction (ddPCR) method for the simultaneous detection of S. pneumoniae, M. pneumoniae and H. influenzae DNA. Specific primers and probes were designed for ddPCR. The sensitivity and specificity of the ddPCR assay were evaluated using standard strains, positive samples and 26 common pathogenic bacteria. One hundred and sixty-seven clinical samples were collected and tested via ddPCR, qPCR, bacterial culture and microfluidic chip technology. The limits of detection (LoDs) of ddPCR were 2.5, 2.8 and 2.0 copies/μL for S. pneumoniae, M. pneumoniae and H. influenzae, respectively, which were approximately tenfold lower than the LoDs of qPCR. For 167 clinical samples, the positivity rates of ddPCR and microfluidic chip for S. pneumoniae and M. pneumoniae were 27.5% and 22.8%, respectively, which were higher than those of qPCR 25.7% and 21.6%. The positive rate of H. influenzae detection via ddPCR and microfluidic chip method was 29.9%, which was higher than that of qPCR (28.7%). The clinical sensitivity for S. pneumoniae, M. pneumoniae and H. influenzae improved from 97.4%, 94.7% and 95.1% for qPCR to 100% for ddPCR. Moreover, ddPCR showed less inhibition by the inhibitor in respiratory specimens than qPCR. The multiplex ddPCR assay established in this study can accurately detect S. pneumoniae, M. pneumoniae and H. influenzae DNA and can be used as an auxiliary tool for the clinical identification of pathogens and guidance of antibiotic therapy.

Timely and accurate diagnosis is essential for the effective treatment of Haemophilus influenzae (HI) infection. Notably, a rapid point-of-care testing (POCT) method for the diagnosis of HI DNA is still lacking. The aim of this study was to establish a one-pot detection device for HI DNA detection using CRISPR/Cas13a that would be applicable for POCT. We established a two-step polymerase chain reaction (PCR)-CRISPR assay, a two-step recombinase-aided amplification (RAA)-CRISPR assay, and a one-pot RAA-CRISPR assay. Additionally, reagent lyophilization, rapid lysis technology, and a one-pot detection device were integrated to construct an extraction-free one-pot detection device, named EFORCA (extraction-free one-pot RAA-CRISPR/Cas13a assay). Validation was performed on 90 simulated samples and 77 clinical samples. The two-step PCR/RAA-CRISPR assay for HI DNA detection was established and optimized, with a detection limit of 1 copy/μL and 100% specificity. The sensitivity and specificity of the two-step PCR-CRISPR assay for the 90 simulated clinical samples were 96.7% and 95%, respectively, and for the 77 clinical samples, 97.5% and 100%, respectively. The one-pot RAA-CRISPR assay and the EFORCA detected 50 and 25 CFU/mL HI in bacterial suspensions within 30 min, respectively. The valuation results for 77 clinical samples demonstrated that the sensitivity of both methods was 95%, which was greater than the sensitivity of quantitative real-time PCR (qPCR) (92.5%) with the same specificity of 100%. We developed an extraction-free one-pot RAA-CRISPR/Cas13a assay for HI detection, which effectively performs a POCT test and is a valuable tool for the early detection and monitoring of HI infection.IMPORTANCETimely and accurate diagnosis is essential for the effective treatment of Haemophilus influenzae (HI) infection. Notably, a rapid point-of-care testing (POCT) method for the diagnosis of HI DNA is still lacking. In this study, sensitive two-step polymerase chain reaction (PCR)-CRISPR and recombinase-aided amplification (RAA)-CRISPR assays were developed, followed by the establishment of the one-pot RAA-CRISPR assay through integration of RAA, CRISPR, and rapid lysis for HI detection. Additionally, reagent lyophilization, rapid lysis technology, and a one-pot detection device were integrated to construct the extraction-free one-pot RAA-CRISPR/Cas13a assay (EFORCA). With its high sensitivity, specificity, rapid turnaround time, and operational simplicity, this assay shows great potential as a practical diagnostic tool for HI infection in small laboratory settings.

Epsilon toxin (ETX), a potential agent of biological and toxic warfare, causes the death of many ruminants and threatens human health. It is crucial to understand the toxic mechanism of such a highly lethal and rapid course toxin. In this study, we detected the effects of ETX on the proteome and phosphoproteome of MDCK cells after 10 min and 30 min. A total of 44 differentially expressed proteins (DEPs) and 588 differentially phosphorylated proteins (DPPs) were screened in the 10 min group, while 73 DEPs and 489 DPPs were screened in the 30 min group. ETX-induced proteins and phosphorylated proteins were mainly located in the nucleus, cytoplasm, and mitochondria, and their enrichment pathways were related to transcription and translation, virus infection, and intercellular junction. Meanwhile, the protein–protein interaction network screened out several hub proteins, including SRSF1/2/6/7/11, SF3B1/2, NOP14/56, ANLN, GTPBP4, THOC2, and RRP1B. Almost all of these proteins were present in the spliceosome pathway, indicating that the spliceosome pathway is involved in ETX-induced cell death. Next, we used RNAi lentiviruses and inhibitors of several key proteins to verify whether these proteins play a critical role. The results confirmed that SRSF1, SF3B2, and THOC2 were the key proteins involved in the cytotoxic effect of ETX. In addition, we found that the common upstream kinase of these key proteins was SRPK1, and a reduction in the level of SRPK1 could also reduce ETX-induced cell death. This result was consistent with the phosphorylated proteomics analysis. In summary, our study demonstrated that ETX induces phosphorylation of SRSF1, SF3B2, THOC2, and SRPK1 proteins on the spliceosome pathway, which inhibits normal splicing of mRNA and leads to cell death.

Disease X stands as a symbol for a subset of emerging infectious diseases rising to prominence as a significant challenge to global health security. This comprehensive review delves into the historical context, precise definition, and illustrative case studies of disease X, including notable examples, such as COVID-19, SARS, and Ebola. The discussion then transitions to an examination of the potential origins of disease X, with a particular focus on zoonotic pathways and the influence of environmental factors. The review concludes with a suite of proposed strategies aimed at the prevention and containment of disease X, emphasizing the critical role of vigilant surveillance, swift response mechanisms, and the necessity of fostering international cooperation.

OBJECTIVES:Indwelling urinary catheter is closely associated with the occurrence of urinary tract infection (UTI). Herein, we further explored the correlation of urinary catheter indwelling time and UTI. METHODS:Retrospectively, the medical data of nosocomial patients (n = 681) were collected during two quarters of April 2023 to June 2023 (the second quarter, 23.4-23.6, n = 330) and July 2023 to September 2023 (the third quarter, 23.7-23.9, n = 351). The baseline data and incidence of catheter-related UTI were analysed. The total hospitalisation days and indwelling urinary catheter days of patients in five departments were assessed, namely, coronary care unit (CCU), respiratory intensive care unit (RICU), surgical intensive care unit (SICU), neurology intensive care unit (NICU) and cardiac surgical intensive care unit (CSICU) departments. The correlation between hospitalisation days/indwelling urinary catheter days and the occurrence of UTI was evaluated by Spearman correlation analysis. RESULTS:In the CCU, RICU, SICU, NICU and CSICU departments, the number of patients was 463, 83, 29, 91 and 15, respectively. During 23.4-23.6, the incidence of catheter-associated UTI (CAUTI) was 0, 2.85, 6.12, 0 and 12.99 per 1000 urinary catheter days in CCU, RICU, SICU, NICU and CSICU, respectively. During 23.7-23.9, the incidence of CAUTI was 2.98, 6.13, 8.66, 0 and 0 per 1000 urinary catheter days in CCU, RICU, SICU, NICU and CSICU, respectively. Notably, hospitalisation days/indwelling urinary catheter days were positively correlated with the occurrence of CAUTI in each quarter (p < 0.05). CONCLUSIONS:There was a positive correlation between urinary catheter indwelling time and the occurrence of UTI.

N95 respirators are the core equipment used by healthcare workers to prevent the spread of respiratory diseases. The protective effect of N95 against infection spread depends on the fit of the N95 to the wearer, which is related to the wearer's facial dimensions. The purpose of this cross-sectional study was to assess the relationship between the fit of three types of N95 and facial dimensions. A total of 305 healthcare workers from ten hospitals in Beijing were recruited for this study. Facial dimensions of workers were measured using Intel RealSense Depth Camera D435. Fit testing was conducted on three types of N95 using the TSI-8038 Porta Count Pro + Respirator Fit Tester. Possible associations between the fit test results and facial dimension data were examined. A Porta Count reading of 100 was used as the criterion for an acceptable fit. The fit of the folding respirators was positively correlated with nose length (r = 0.13, p = 0.02), nose height (r = 0.14, p = 0.02), and face width (r = 0.12, p = 0.03), whereas that of flat respirators was correlated with nose width (r = 0.16, p < 0.01), chin length (r = 0.18, p < 0.01), and pro-face width (r = 0.13, p = 0.02), and that of arched respirators was correlated with the nose length (r = 0.13, p = 0.03). The fit of N95 for wearers depends on their facial features. The results of this study can provide advice for medical workers to choose the appropriate N95. Medical staff should fully consider their facial dimensions when choosing an appropriate N95 to improve the protective efficacy of respirators and to reduce the risk of infection by respiratory diseases.

BACKGROUND/PURPOSE:Carbapenem-resistant Klebsiella pneumoniae (CRKP) is capable of causing serious community and hospital-acquired infections. However, currently, the identification of CRKP is complex and inefficient. Hence, this study aimed to develop methods for the early and effective identification of CRKP to allow reasonable antimicrobial therapy in a timely manner. METHODS:K. pneumoniae (KP)-, K. pneumoniae carbapenemase (KPC)- and New Delhi metallo-β-lactamase (NDM)- specific CRISPR RNAs (crRNAs), polymerase chain reaction (PCR) primers and recombinase-aided amplification (RAA) primers were designed and screened in conserved sequence regions. We established fluorescence and lateral flow strip assays based on CRISPR/Cas13a combined with PCR and RAA, respectively, to assist in the detection of CRKP. Sixty-one clinical strains (including 51 CRKP strains and 10 carbapenem-sensitive strains) were collected for clinical validation. RESULTS:Using the PCR-CRISPR assay, the limit of detection (LOD) for KP and the blaKPC and blaNDM genes reached 1 copy/μL with the fluorescence signal readout. Using the RAA-CRISPR assay, the LOD could reach 101 copies/μL with both the fluorescence signal readout and the lateral flow strip readout. Additionally, the positivity rates of CRKP-positive samples detected by the PCR/RAA-CRISPR fluorescence and RAA-CRISPR lateral flow strip methods was 92.16% (47/51). The sensitivity and specificity reached 100% for KP and blaKPC and blaNDM gene detection. For detection in a simulated environmental sample, 1 CFU/cm2 KP could be detected. CONCLUSION:We established PCR/RAA-CRISPR assays for the detection of blaKPC and blaNDM carbapenemase genes, as well as KP, to facilitate the detection of CRKP.

目的 系统评价正念瑜伽团体心理干预改善孕期焦虑及分娩恐惧的疗效.方法 计算机检索Cochrane Library、PubMed、Embase、CBM、CNKI、VIP、万方数据库中公开发表的孕期进行正念瑜伽团体心理治疗的随机对照研究.对符合纳入标准的文献,用RevMan 5.3软件进行Meta分析,对比两组的疗效.结果 共纳入5项研究,共计422人.运用状态-特质焦虑量表(state-trait anxiety inventory,STAI)对干预和对照组进行了评估,正念瑜伽团体心理干预后干预组患者的STAI-S及STAI-T分数均较对照组显著降低[WMD =-7.81,95％CI(-12.92,-2.70),P = 0.003]、WMD =-5.77,95％CI(-11.84,-0.30),P= 0.06],焦虑自评量表评分较对照组显著降低[WMD =-15.68,95％CI(-17.40,-13.96),P＜0.001],分娩恐惧量表评分较对照组显著降低[WMD=-12.08,95％CI(-14.82,-9.34),P＜0.001].结论 正念瑜伽团体心理干预可缓解孕期的焦虑水平,降低孕妇分娩恐惧.

目的 了解北京市西城区各区属医院入院患者多重耐药菌(multidrug-resistant organisms,MDROs)筛查情况.方法 将北京市西城区十家区属医院,在2015年6月至2016年9月期间的入院患者,依患者来源,分为社区入院组(来自医院周边社区)和医院转入组(由区域内三甲医院出院后转入).入院后48 h内采集鼻拭子和肛拭子,利用产色鉴定培养基技术,进行几种常见MDROs快速筛查,了解入院患者MDROs的筛查阳性比例、菌种分布,以及比较不同来源患者间的差异.结果 研究期间共纳入644例患者,其中医院转入组304例,占47.2％.医院转入组中至少一个部位一种细菌筛查阳性的患者比例,显著高于社区入院组(72.4％vs 51.5％,P<0.001).鼻拭子筛查的阳性率,医院转入组同样高于社区入院组(11.8％vs 3.2％,P<0.001),以耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)阳性率最高.两组患者肛拭子的阳性率均高于鼻拭子,而肛拭子2种及以上细菌筛查阳性的患者,医院转入组也高于社区入院组(18.9％vs 12.8％,P=0.046),以大肠埃希菌(Escherichia coli,ECO)、克雷伯菌-肠杆菌-沙雷菌-柠檬酸杆菌(Klebsiella-Enterobacter-Serratia-Citrobacter,KESC)阳性率最高.既往脑血管疾病、高血压、入院前4周内手术或介入治疗史、抗菌药物暴露史及入院时存在SIRS,是入院患者MDROs筛查阳性的危险因素.结论 北京市西城区各区属医院入院患者MDROs筛查阳性率高,以区域内医院转入患者为著.应引起院感管理部门的高度重视.

Objective:To learn the application of nosocomial infection prevention and control measures as stipulated in COVID-19 emergency plans by medical institutions at all levels in the region, for the purpose of strengthening epidemic prevention and control.Methods:During March 12-13, 2020, customized questionnaires were used to learn from 186 hospitals and medical institutions regarding the basics of their nosocomial prevention management departments, emergency plan application and revisions made. Comparison of the ratios or constituent ratios were tested with χ2 test, while the continuous variables analysis between groups was verified with one-way ANOVA. Results:77.53% of the medical institutions had set up independent nosocomial infection management departments, and 87.30% of the institutions were qualified. 80% of the medical institutions had in place emergency plans for respiratory infectious diseases, but 98.05% of them had revised their plans during the pandemic, with an average of 10.85 newly added and revised provisions. Only 30.11% of emergency planed provide for clearly graded early warning.Conclusions:Efforts should be upgraded to develop an emergency prevention and control system for infection prevention and control in epidemics, and improve technical support for infection prevention and control in the system; to strengthen the clearly-graded early warning and graded responses in a scientific manner; and conduct regular drills, revise plan to ensure its applicability.

OBJECTIVES:Most Chinese hospitals have customized environmental cleaning policies and systems, with limited data availability based on evidence-based medicine. This study investigated the relationship between multidrug-resistant organism (MDRO) colonization in intensive care unit (ICU) patients and ICU surface bacterial contamination status.METHODS:This cross-sectional study comprised MDRO screening in ICU patients using bacterial cultivation by chromogenic medium; samples were collected before (control group) and after implementation of enhanced cleaning (cleaning group). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used to identify and analyze microorganisms; the relationships of MDRO colonization with infection and environmental bacteria were analyzed.RESULTS:In total, 196 patients were enrolled in the study (104 and 92 in control and cleaning groups, respectively); 1042 MDROs were subjected to screening before and after cleaning. After cleaning, the rate of MDRO detection on surfaces of frequently touched objects in ICUs decreased from 31.77% to 13.32%. There were fewer MDRO homologues in the cleaning group than in the control group. Moreover, the cleaning group had a shorter ICU stay and significantly lower mortality rate.CONCLUSIONS:Enhanced environmental cleaning and disinfection could reduce environmental MDRO accumulation and suppress MDRO colonization in ICUs, thereby reducing nosocomial infections and improving adverse patient outcomes.

目的:通过对医院后勤工作范围、工作特点进行分析,探讨新冠肺炎疫情期间加强医院后勤工作管理、降低医院感染风险的措施.方法:对医院后勤各部门存在的感染风险进行评估,制定规范的医院后勤工作管理制度与新冠肺炎防控工作流程,按照人员暴露风险实行分级防护,加强对后勤人员的培训和对后勤工作的监督检查.结果:新冠肺炎疫情期间,规范医院后勤工作制度和工作流程,做好后勤人员个人防护,对后勤工作人员加强院感知识培训和院感防控工作监督指导,能有效预防和控制医院感染的发生.结论:新冠肺炎疫情期间,通过实施各种管理措施,加强医院后勤工作感染风险管控,并持续改进,有助于降低医院感染风险.

Objective:To understand the problems in using the personal protective equipment (PPE) by medical staffs working in the ward of coronavirus disease 2019 (COVID-19), so as to provide the reference on optimizing PPE wearing and taking-off process and accelerating technical innovations and standard formulations.Methods:A unified questionnaire was designed by Wenjuanxing Platform. The survey applied "web-based survey + on-site survey" method to investigate PPE usage by 222 health care workers working in the ward of COVID-19.Results:Among the medical staffs, 61.26% (136/222) had no doubt about PPE wearing process. Three most problematic protective equipments were medical respirator (48, 21.62%), goggles or face shield (29, 13.06%) and shoe/boot covers (26,11.71%). There were no statistically significant differences in gender, time of service or occupation on the doubts of protective equipments ( P>0.05). The problems in PPE wearing and taking-off process were mainly leakage checking in wearing medical respirator (102, 45.95%), taking off integrated protective clothing while rolling, difficulties in taking off the clothing from shoulders (81, 36.49%), difficulties in taking off both the clothing and outer gloves together (76, 34.23%) and contamination of face while taking off medical respirator (34, 15.32%), and so on. Conclusions:PPE should be chosen and used scientifically. Training on PPE wearing and taking-off process should be strengthened on the basis of identifying the key elements in the processes. Technical innovation and standards for PPE should be accelerated to reduce the risk of infection effectively for the medical workers.

新型冠状病毒肺炎(COVID-19)是一种由新型冠状病毒(SARS-CoV-2)引起的急性传染病,其出现及其传播给全球公共卫生安全带来了极大的挑战.COVID-19定点收治医疗机构是收治确诊患者,特别是重症和危重症患者的前沿阵地.如何快速建立针对疫情的医院感染防控体系,降低医疗机构内传播的风险,保证一线医务人员、其他员工与来院就诊的其他患者及家属免遭感染,同时保证新型冠状病毒感染确诊患者诊疗救治工作的顺利开展,是很多医疗机构都会直接面对的问题.首都医科大学附属北京佑安医院作为北京市级定点收治新型冠状病毒肺炎患者医疗机构之一,通过总结和归纳其疫情期间的具体对策,以探索医院感染防控策略.

目的 探讨快速设置针对新型冠状病毒肺炎(coronavirus disease 2019,COVID-19)疫情的医院感染防控体系,在开展诊疗救治工作的同时有效阻断新型冠状病毒在院内的感染风险.方法 从传染病管理三要素角度,以及收治COVID-19患者期间执行的医院感染防控策略,包括人员、物资、流程布局的管理等方面进行总结分析.结果 急诊行手术的患者经过有效的筛查及合理的分级、分区管理,严格执行防控动线及消毒隔离措施并进行核查.在完成诊疗救治工作的同时,未发生医护人员及住院患者的院内感染事件.结论 科学有效的执行防控策略,可以在COVID-19流行期间,在承担COVID-19患者收治任务同时,正常收治其他患者,并预防其在院内的传播流行.

Objective:To compare the efficacy of a new compound peracetic acid disinfectant with glutaraldehyde in killing residue viruses and bacteria of digestive endoscopes used by chronic hepatitis B, hepatitis C and AIDS patients with high viral load.Methods:Twenty-four patients with HBV-DNA ≥10 5 IU/ml, HCV-RNA ≥10 5 IU/ml or human immunodeficiency virus (HIV)-RNA ≥10 4 copies/ml were randomly divided into the experimental group (the new compound peracetic acid disinfection group) or the control group (glutaraldehyde disinfection group) according to the diseases. HBV-DNA, HCV-RNA, HIV-RNA, bacterial colony and adenosine triphosphate (ATP) bioluminescence on the digestive endoscopes they used were measured in four different point of time. Results:No HBV-DNA, HCV-RNA and HIV-RNA was detected on the inner and outer surface of digestive endoscopes after disinfection in the experimental group and the control group. There was no significant difference in bacterial colony between the experimental group and the control group ( Z=0.718, P=0.473). The qualified rate of disinfection was 100%. There was no significant difference in ATP contents before and after disinfection between the experimental group and the control group ( Z=1.334, P=0.182). The adverse reactions of the cleaning personnel to the disinfectant in the experimental group were less than those in the control group. Conclusions:The new compound peracetic acid disinfectant and glutaraldehyde can reach the requirements of the national standard of disinfection. The time of treatment with the new compound peracetic acid disinfectant was shorter with less adverse effect on the cleaning personnel.

OBJECTIVE: To rapidly establish a prevention and control system in response to the epidemic so as to reduce the risk of infection in medical staff during diagnosis and treatment of patients with confirmed COVID-19, other patients and their family members METHODS: The strategies for prevention and control of nosocomial infection that were carried out during the treatment of the COVID-19 patients, including personnel management, material management, process management and execution supervision, were summarized and analyzed RESULTS: The working group on prevention and control of the epidemic ran efficiently, the classification of the personnel in medical institutions, classification and division management, scientific assessment of usage amount of protective equipment, reasonable allocation and surveillance of epidemiological history and symptoms covered all the patients and working staff of the entire hospital, the construction was combined with the working process, the control line was effectively implemented, and the status of implementation of the measures was rigidly checked out CONCLUSION: The scientific implementation of the prevention and control strategies maintains orderly diagnosis and treatment of the patients with COVID-19 and effectively prevent hospital-acquired COVID-19

目的 评价一种新型纳米长效消毒液对耐碳青霉烯病原菌的抑菌效果.方法 选取耐碳青霉烯类抗菌药物的肺炎克雷伯菌(carbapenem resistant Klebsiella pneumonia,CR-KPN)、鲍曼不动杆菌(carbapenem resistant Acinetobacter baumannii,CRAB)、铜绿假单胞菌(carbapenem resistant Pseudomonas aeruginosa,CRPA)各5株,试验组使用新型纳米长效消毒液,对照组分别使用主要成分为复合双链季胺盐的消毒液和不同浓度的复方含氯消毒液,制备消毒剂纸片并贴于接种细菌的平皿表面,37℃培养24 h后测量抑菌圈平均直径,判断抑菌效果及抑菌时效.结果 试验组在24 h内对CR-KPN和CRAB的抑制效果优于各对照组,抑菌圈平均直径分别为(17.88±0.95)和(17.48±0.61)mm,与各对照组相比差异均有统计学意义,且抑制作用至少持续3d;试验组与对照组对CRPA均无抑制作用.结论 新型纳米消毒液对CR-PA无抑制作用,对CR-KPN和CRAB的抑制效果较好,但随着时间的延续,抑制效果呈下降趋势.

目的 分析2014-2018年金黄色葡萄球菌(Staphylococcus aureus,S.A)的临床分布特征和耐药情况,为临床干预与研究提供依据.方法 收集2014年1月1日至2018年12月31日首都医科大学附属北京佑安医院的S.A临床分离株,采用自动化仪器法或纸片扩散法按统一方案进行细菌药物敏感性试验,应用WHONET 5.6和SPSS 19.0软件进行统计分析.结果 5年共检出S.A 387株,占全部检出菌的4.2％,耐甲氧西林金黄色葡萄球菌(methicillin-resistant staphylococcus aureus,MRSA)平均检出率为31.7％.分离出S.A菌株最多的标本来自呼吸道(118株)、血液(102株)和腹水(61株),占全部标本分离菌株的72.7％.S.A对青霉素的耐药率最高,为89.9％,其次是红霉素57.2％、苯唑西林29.4％、克林霉素27.0％和四环素23.9％,仅在2018年检出1株利奈唑胺耐药菌株,尚未检出对万古霉素、替考拉宁耐药的菌株.结论 2014-2018年我院S.A分离率、MRSA检出率和耐药性均低于全国平均水平,但呈增长趋势,应予以重视,进一步采取有效控制措施控制,避免医院感染.