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Mengjie Hu,1,* Songling Chai,2,* Wenyi Lei,1 Kexin Liao,1 Rongxin Zhang1 1Department of Dermatology, The Second Hospital of Dalian Medical University, Dalian, Liaoning, People’s Republic of China; 2Department of Oral and Maxillofacial Surgery, Stomatology College of Dalian Medical University, Dalian, Liaoning, People’s Republic of China*These authors contributed equally to this workCorrespondence: Rongxin ZhangDepartment of Dermatology, The Second Hospital of Dalian Medical University, No. 467 Zhongshan Road, Dalian, 116027, Liaoning, People’s Republic of ChinaTel +8617709873080Fax +86-411-84672130Email zrx_1980@126.comAbstract: Cutaneous cryptococcosis, an infectious disease resulting from Cryptococcus neoformans, primarily affects immunodeficient individuals. Here, we report a case of mediastinal small cell carcinoma (MSCC) complicated with multiple skin and soft tissue infections mimicking erysipelas and cellulitis. Antibiotics for bacteria were ineffective and a culture of pus from the infected areas revealed Cryptococcus neoformans in this patient. The absence of any evidence indicative of systemic cryptococcal infection leads to a final diagnosis of primary cutaneous cryptococcosis (PCC). Following two weeks of fluconazole at 400 mg/day and 200 mg/day for the subsequent three months, combined with incision, irrigation and drainage, the wound gradually healed. An analysis and discussion of the clinical features of this patient are presented. This case alerts clinicians as to the possibility of Cryptococcus neoformans in patients with advanced malignant tumors complicated with multiple skin and soft tissue infections. While a timely diagnosis and treatment of PCC in this patient resulted in a favorable outcome, the patient succumbed to the malignant tumor at six months post-discharge.Keywords: mediastinal small cell carcinoma, primary cutaneous cryptococcosis, Cryptococcus neoformans, immunodeficiency, therapy

Qi Wang,1 Zhanwei Wang,1 Feifei Zhang,1 Chunjiang Zhao,1 Bin Yang,2 Ziyong Sun,3 Yaning Mei,4 Feng Zhao,5 Kang Liao,6 Dawen Guo,7 Xiuli Xu,8 Hongli Sun,9 Zhidong Hu,10 Yunzhuo Chu,11 Yi Li,12 Ping Ji,13 Hui Wang1 1Department of Clinical Laboratory, Peking University People’s Hospital, Beijing 100044, People’s Republic of China; 2Department of Clinical Laboratory, The First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, People’s Republic of China; 3Department of Clinical Laboratory, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, People’s Republic of China; 4Department of Clinical Laboratory, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, People’s Republic of China; 5Department of Infectious Diseases, Sir Run Shaw Hospital (SRRSH), Affiliated with the Zhejiang University School of Medicine, Hangzhou 310016, People’s Republic of China; 6Department of Laboratory Medicine, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, People’s Republic of China; 7Department of Clinical Laboratory, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, People’s Republic of China; 8Department of Clinical Laboratory, Xijing Hospital of Air Force Military Medical University, Xi’an 100191, People’s Republic of China; 9Department of Clinical Laboratory, Peking Union Medical College Hospital, Beijing 100730, People’s Republic of China; 10Department of Clinical Laboratory, General Hospital of Tianjin Medical University, Tianjin 300052, People’s Republic of China; 11Department of Clinical Laboratory, The First Affiliated Hospital of China Medical University, Shenyang 110001, People’s Republic of China; 12Department of Clinical Laboratory, Henan Provincial People’s Hospital, Zhengzhou 450003, People’s Republic of China; 13Department of Clinical Laboratory, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, People’s Republic of ChinaCorrespondence: Hui WangDepartment of Clinical Laboratory, Peking University People’s Hospital, Beijing 100044, People’s Republic of ChinaTel/ Fax +86-10-88326300Email whuibj@163.comPurpose: The Chinese Meropenem Surveillance Study (CMSS) was conducted every 2 years from 2010 to 2018 to monitor the antimicrobial activity of commonly used antimicrobial agents against nosocomial gram-negative bacilli in China.Methods: From 2010 to 2018, 6,537 gram-negative bacilli were collected from 14 teaching hospitals. The minimum inhibitory concentrations (MICs) of meropenem and other antimicrobial agents were determined using the agar dilution and broth microdilution methods.Results: Continuous surveillance indicated that, except for Klebsiella pneumoniae, the susceptibility of Enterobacterales to carbapenems was relatively stable over time. Carbapenems had the highest activity against the tested isolates, with MIC90 values (MIC for 90% of organisms) ranging from 0.032 mg/L to 8 mg/L. More than 90% of bacteria were susceptible to either meropenem or imipenem; more than 80% were susceptible to ertapenem. The prevalence of extended-spectrum beta-lactamase (ESBL)-producing E. coli, K. pneumoniae, and P. mirabilis each year was 50.4– 64.3%, 18– 41.2%, and 1.9– 33.8%, respectively. The prevalence of carbapenem-resistant K. pneumoniae (CRKP) and carbapenem-resistant Acinetobacter baumannii (CRAB) continued to increase significantly over time, from 7.6% to 21.2% and 64.6% to 69.3%, respectively. The prevalence of CRKP was higher from urinary tract infections (25.4%) than from bloodstream infections (14.2%), intra-abdominal infections (14.5%), and respiratory infections (14.4%). In total, 129 CRKP isolates were evaluated by PCR; of these, 92 (71.3%) carried the blaKPC-2 gene. Colistin maintained very high in vitro antimicrobial activity against P. aeruginosa and A. baumannii (more than 95% of isolates exhibited susceptibility at all timepoints).Conclusion: The results indicate an increase in K. pneumoniae resistance to carbapenems over time, mainly owing to KPC-type carbapenemase production. A. baumannii was severely resistant to carbapenems in China. Ongoing MIC-based resistance surveillance, like CMSS, provides additional data for clinical anti-infective treatment.Keywords: CMSS, gram-negative bacilli, antimicrobial susceptibility surveillance, carbapenem-resistant

Lan-Lan Zhong,1,2,* Qian Zhou,3,* Cui-Yan Tan,3,* Adam P Roberts,4,5 Mohamed Abd El-Gawad El-Sayed Ahmed,1–2,6 Guanping Chen,1,2 Min Dai,7 Fan Yang,8 Yong Xia,9 Kang Liao,10 Yingjian Liang,3 Yongqiang Yang,1,11 Siyuan Feng,1,2 Xiaobin Zheng,3,* Guo-Bao Tian1,2,*1Program in Pathobiology, The Fifth Affiliated Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangdong 519000, People’s Republic of China; 2Ministry of Education, Key Laboratory of Tropical Diseases Control, Sun Yat-sen University, Guangzhou 510080, People’s Republic of China; 3Department of Respiratory Medicine, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, People’s Republic of China; 4Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, L3 5QA, UK; 5Centre for Drugs and Diagnostics, Liverpool School of Tropical Medicine, Liverpool L3 5QA, UK; 6Department of Microbiology and Immunology, Faculty of Pharmaceutical Sciences and Drug Manufacturing, Misr University for Science and Technology (MUST), Cairo, Egypt; 7School of Laboratory Medicine, Chengdu Medical College, Chengdu 610500, People’s Republic of China; 8Department of Microbiology, School of Basic Medical Science, Xinxiang Medical University, Xinxiang 453003, People’s Republic of China; 9Department of Clinical Laboratory Medicine, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, People’s Republic of China; 10Department of Clinical Laboratory, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, People’s Republic of China; 11School of Pharmaceutical Sciences (Shenzhen), Sun Yat-sen University, Guangzhou 510006, People’s Republic of China*These authors contributed equally to this workPurpose: The discovery of the plasmid-mediated colistin resistance genes, mcr, revealed a mechanism of transmission of colistin resistance, which is a major, global public health concern especially among individuals infected with carbapenem-resistant Gram-negative bacteria. To monitor the spread and epidemiology of mcr genes, a convenient and reliable method to detect mcr genes in clinical isolates is needed, especially in the primary care institutions. This study aimed to establish a restriction endonuclease-based multiplex loop-mediated isothermal amplification (multi-LAMP) assay to detect mcr genes (mcr-1 to mcr-5) harbored by colistin-resistant bacteria.Methods: A triple-LAMP assay for mcr-1, mcr-3, and mcr-4 and a double-LAMP assay for mcr-2 and mcr-5 were established. The sensitivity and specificity of the LAMP reactions were determined via electrophoresis and visual detection.Results: The sensitivity of the LAMP assay was 10-fold greater than that of PCR, with high specificity among the screened primers. Specific mcr genes were distinguished in accordance with band numbers and the fragment length of the digested LAMP amplification products. Furthermore, the LAMP assay was confirmed as a rapid and reliable diagnostic technique upon application for clinical samples, and the results were consistent with those of conventional PCR assay.Conclusion: The multi-LAMP assay is a potentially promising method to detect mcr genes and will, if implemented, help prevent infections by drug-resistant bacteria in primary-care hospitals due to rapid and reliable surveillance. To our knowledge, this is the first study to report the application of LAMP to detect mcr-2 to mcr-5 genes and the first time that multi-LAMP has been applied to detect mcr genes.Keywords: mcr genes, colistin resistance, multi-LAMP, rapid detection, enzyme digestion

Fan Yang,1,* Cong Shen,2,3,* Xiaobin Zheng,4 Yan Liu,5 Mohamed Abd El-Gawad El-Sayed Ahmed,2,3,6 Zihan Zhao,2,3 Kang Liao,7 Yaling Shi,8 Xin Guo,2,3 Ruoxuan Zhong,2,3 Zhimin Xu,2,3 Guo-Bao Tian2,3 1Department of Microbiology, School of Basic Medical Science, Xinxiang Medical University, Xinxiang, China; 2Department of Microbiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China; 3Key Laboratory of Tropical Diseases Control (Sun Yat-sen University), Ministry of Education, Guangzhou, China; 4Department of Respiratory Medicine, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, China; 5Department of Laboratory, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, China; 6Department of Microbiology and Immunology, Faculty of Pharmacy, Misr University for Science and Technology (MUST), Cairo, 6th of October City, Egypt; 7Department of Clinical Laboratory, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China; 8Department of Clinical Laboratory, Guangzhou Eighth People’s Hospital, Guangzhou Medical University, Guangzhou, China *These authors contributed equally to this work Background: As a result of the growing prevalence of the plasmid-mediated mobile colistin resistance gene mcr-1 among Gram-negative bacteria, the surveillance of mcr-1 has been globally applied. In our study, we aimed to shed light on the possibility of transmission of mcr-1-resistant isolates through market retail fruits. Methods and results: Herein, 133 different fruit surface samples were collected and screened for the different MCR variants (mcr-1 to mcr-8) using PCR and confirmed with sequencing. We identify for the first time mcr-1-carrying Escherichia coli and Klebsiella pneumoniae from market retail fruits in Guangzhou, China. Minimum inhibitory concentrations were detected by the broth microdilution method. Liquid mating was performed to check the transferability of the mcr-1 gene. Pulsed field gel electrophoresis analysis of S1 nuclease-digested DNA and Southern blotting were performed to check the location of the mcr-1 gene. Then, whole-genome sequencing and in silico multilocus sequence typing analysis were performed. Conclusion: We showed that E. coli GB110 can mediate the spreading of antibiotic resistance genes through the food chain, while K. pneumoniae GB015 was considered to be the progenitor of the most successful multidrug-resistant clone. Since fruits are usually consumed fresh, this may serve as a direct source of mcr-1-producing bacteria in humans that requires prompt surveillance and intervention to limit the spread of resistance. Keywords: colistin, mcr-1, Escherichia coli, Klebsiella pneumoniae, fruit

Yujian Liang,1,&ast; Chenfeng Zhao,2,&ast; Yuhang Lu,3,&ast; Kang Liao,2 Yannan Kong,4 Mengzhi Hong,2 Liubing Li,2 Yili Chen2 1Department of Pediatric Intensive Care Unit, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, 510080, People’s Republic of China; 2Department of Laboratory Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, 510080, People’s Republic of China; 3Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong, 510080, People’s Republic of China; 4Department of Laboratory Medicine, the Nanhai’s Fifth People’s Hospital of Foshan, Foshan, Guangdong, 528231, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yili Chen; Liubing Li, Department of Laboratory Medicine, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, 510080, People’s Republic of China, Email chenyli3@mail.sysu.edu.cn; lilb8@mail.sysu.edu.cnBackground: Carbapenem-resistant Enterobacteriaceae (CRE) is rapidly becoming a major threat to hospitalized children worldwide. The purpose of this study was to summarize etiological characteristics and identify risk factors relevant to CRE bloodstream infection (BSI) and short-term mortality among pediatric patients in China.Methods: In this study, we included 370 inpatients ≤ 17 years old with BSI caused by Enterobacteriaceae in China from January 2013 to December 2022. By collecting data on demographics, etiological features, and clinical outcomes, we conducted an in-depth analysis.Results: Among the 370 BSI patients with infections caused by Enterobacteriaceae, 35 patients (9.46%) were caused by CRE. Among these CRE strains, Klebsiella pneumoniae (49.46%) was the most important pathogen of BSI in pediatric patients, followed by Escherichia coli (31.62%) and Enterobacter cloacae (5.95%). The most frequent carbapenemase was NDM (23/35, 65.71%), followed by KPC (8/35, 22.86%). The overall 28-day mortality rate of children with an Enterobacteriaceae BSI episode was 1.89% (7/370), of which CRE BSI patients (3/35, 8.57%) were significantly higher than CSE patients (4/335, 1.19%, P < 0.001). Congenital malformation (OR: 8.162, 95% CI: 3.859– 16.680, P < 0.001) and catheter-related (OR: 6.645, 95% CI: 3.159– 13.28, P: < 0.001) were associated with the development of CRE BSI in pediatric patients. A multivariate analysis showed that the infection of CRE (OR 7.758, 95% CI 1.869– 29.62, P = 0.021) were independent risk factors for 28-day mortality of Enterobacteriaceae BSI. When the MIC of any carbapenems was ≥ 8 μg/mL, the mortality rate in the ICU was higher (P < 0.05).Conclusion: Congenital malformation, previous cephalosporin/carbapenems administration, and catheter-related conditions were closely related to the development of CRE BSI. The mortality rate of CRE BSI was higher. NDM was the predominant carbapenemase-producing mechanism in children.Keywords: carbapenem-resistant Enterobacteriaceae, bloodstream infection

Xiuxin Tang,1,&ast; Zhiyun Liu,2,&ast; Danni Li,3 Yan Xiong,3 Kang Liao,1 Yili Chen1 1Department of Laboratory Medicine, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong, People’s Republic of China; 2Department of Clinical Laboratory, Huizhou Central People ‘s Hospital, Huizhou, Guangdong, 516001, People’s Republic of China; 3Department of Emergency Medicine, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yili Chen, Department of Laboratory Medicine, The First Affiliated Hospital of Sun Yat-sen University, No. 58 Zhongshan Er Road Guangzhou, Guangzhou, Guangdong Province, 510080, People’s Republic of China, Tel +8620-87330808-8461, Fax +8620-87330808, Email chenyli3@mail.sysu.edu.cnBackground: Elizabethkingia anophelis, an opportunistic pathogen that can cause infections in multiple parts of the human body, has multiple drug resistance and a high mortality rate. However, there have been few reports of infective endocarditis (IE) caused by Elizabethkingia anophelis, which means that diagnosis and treatment face challenges that cannot be ignored. Rapid and accurate identification and drug sensitivity results are needed to make timely treatment adjustments.Case Presentation: An 81-year-old man presented with recurrent fever and increased infection index for more than a month. Based on his clinical symptoms, infection index, reduplicative blood cultures, and results of transesophageal echocardiography, he was ultimately diagnosed with infective endocarditis caused by Elizabethkingia anophelis. The patient had a favorable outcome with a 6-week course of intravenous antibiotic therapy.Conclusion: This is a rare and successfully cured case of IE caused by the pathogen of Elizabethkingia anophelis, which is difficult not only in diagnosis but also in treatment. This case provides a certain referential significance to the treatment of Elizabethkingia anophelis-caused IE in clinical practice.Keywords: Elizabethkingia anophelis, infective endocarditis, drug resistance, treatment