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Linfa Chen,1,2,* Keqi Liao,1,3,* Yutian Zhang,1,3,* Shutao Zheng,1,4 Jiawen He,1,3 Henglei Tang,1,4 Hailing Wu,1,3 Wangtao Zhong,4 Shengnan Li,1,3 You Li1,3 1Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, People’s Republic of China; 2Department of Neurology, Huizhou Third People’s Hospital, Guangzhou Medical University, Huizhou, People’s Republic of China; 3Institute of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, People’s Republic of China; 4Department of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, People’s Republic of China*These authors contributed equally to this workCorrespondence: You Li; Shengnan Li, Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, People’s Republic of China, Email youli805@163.com; 15625102893@163.comBackground: Accumulating evidence suggests that matrix metalloproteinase (MMP) 12 plays a detrimental role in cerebro-cardiovascular diseases, including ischemic stroke (IS). Previous genome-wide association studies (GWAS) correlated the MMP12 rs660599 variant to IS risk in Europeans. However, this association is yet to be elucidated in the Chinese population. This study aims to assess the genetic predisposition of the MMP12 rs660599 G > A variant with regard to IS risk and short-term outcomes in individuals from Southern China.Methods: The Multiplex SNaPshot assay was used to genotype rs660599 in 1035 IS patients and 1061 age-matched healthy controls. Multivariate logistic regression analyses evaluated the effect of the rs660599 G > A polymorphism on IS susceptibility and short-term outcomes.Results: No significant association was found between the rs660599 G > A polymorphism and IS risk, even in dominant and recessive models. However, a relationship between rs660599 genotypes and diabetic status revealed that carriers of the A allele and the GA/AA genotype were more likely to develop IS. The presence of diabetes exacerbated the larger infarct volumes and elevated serum MMP12 levels seen in IS patients with the rs660599 A allele. The A allele of rs660599 and the GA/AA genotype were both correlated to moderate and severe stroke with poor short-term outcomes.Conclusion: The MMP12 rs660599 polymorphism is associated with a higher incidence of IS in people with diabetes and can serve as a biomarker for assessing the severity of IS and its short-term consequences.Keywords: Ischemic Stroke, Matrix Metalloproteinase 12, polymorphism, risk, short-term outcome

Yu-Xin Li,1 Jun Xiong,2 Zheng Zhang,2 Kai Liao,2 Xiao-Hong Zhou,2 Jun Li,2 Jie Xiang,2 Ling-Ling Xu2 1Graduate School, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, People’s Republic of China; 2College of Acupuncture and Massage, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, People’s Republic of ChinaCorrespondence: Jun Xiong, College of Acupuncture and Massage, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, 330004, People’s Republic of China, Email xiongjun196071@163.comBackground: Acupuncture and moxibustion has been applied worldwide in the treatment of various pain diseases including lumbar disc herniation (LDH) and other pain, However, there has been no bibliometric analysis on this aspect in the past five years. Therefore, this study was carried out for finding research trends and fronts in this field using Citespace and VOSviewer.Methods: Publications about acupuncture therapy for LDH were extracted from the Web of Science database and PubMed with an unlimited time frame. A bibliometric analysis and visualization of results was conducted using CiteSpace 6.1.R3 and VOSviewer 1.6.18 on the information of the annual publication, countries, journals, institutions, authors, references, and keywords.Results: A total of 127 publications were included, and the number of publications had increased noticeably over the past 30 years and reached a peak in the past 3 years. The most productive country with the most publications was China, whose Medical University was the institution with the highest volume of publications. The most productive author was Chen Rixin, while the most-cited author was Kreiner DS. Chinese Acupuncture and Moxibustion was the journal with the most publications, and Spine Journal was the most frequently cited journal. In cited references, an article published in The New England Journal of Medicine by Deyo RA had the most citations and the highest centrality. Of the keywords, the five most frequently used keywords include lumbar disc herniation, acupuncture, low back pain, intervertebral disc displacement, and management.Conclusion: Acupuncture and moxibustion can help to relieve symptoms in patients. However, this field is in the early stages of development and requires more high-quality research studies and international collaborations. In addition, exploring the effectiveness and mechanism of acupuncture for LDH is the hot trend in the future.Keywords: acupuncture, lumbar disc herniation, bibliometric analysis, Traditional Chinese Medicine

Rongxin Zhang,1,2 Zhao Hou,2 Kexin Liao,2 Chao Yu,2 Rongrong Jing,1,2 Caixia Tu1,2 1Institute of Integrative Medicine, Dalian Medical University, Dalian, 116044, People’s Republic of China; 2Department of Dermatology, The Second Hospital of Dalian Medical University, Dalian, 116027, People’s Republic of ChinaCorrespondence: Caixia Tu, Department of Dermatology, The Second Hospital of Dalian Medical University, No. 467 Zhongshan Road, Dalian, 116027, People’s Republic of China, Tel +8617709872288, Fax +86 411 84672130, Email tucx2010@sina.comPurpose: Circular RNAs (circRNAs) are abundant, stable, and evolutionarily conserved noncoding RNAs with impacts on cell proliferation, differentiation, invasion, apoptosis, and immunity by acting as an miRNA sponge. This study aimed to investigate the expression of circRNAs in vitiligo and analyze the differentially expressed circRNAs (DEcircRNAs) bioinformatically.Patients and Methods: Biopsies of five lesional and five nonlesional skins of patients with vitiligo and five healthy skins (control) were harvested in this study. The expression profiles of circRNAs and DEcircRNAs were determined by microarray analysis and qRT-PCR. Bioinformatics analysis was used to predict target genes of DEcircRNAs binding to miRNAs and their underlying functions. Meanwhile, a competing endogenous RNA (ceRNA) network was constructed using Cytoscape.Results: A total of 817 and 508 DEcircRNAs were identified in lesional and nonlesional skins of patients with vitiligo, respectively. The results of hsa_circRNA_000957 and hsa_circRNA_101798 validation were consistent with our microarray analysis. Furthermore, 32 miRNA response elements (MREs) and related target genes of DEcircRNAs were identified, whose main functions were involved in the pathogenesis of vitiligo. Hsa_circRNA_000957 and hsa_circRNA_101798 might be candidate biomarkers for vitiligo.Conclusion: This study provides scientific clues for understanding the mechanism of vitiligo.Keywords: vitiligo, circular RNAs, microRNAs, miRNA response element, ceRNA

Zhang W, Liao K, Liu D. Cancer Manag Res. 2020;12:8137–8147. The Editor and Publisher of Cancer Management and Research are retracting the published article. Concerns were raised regarding the duplication of cell apoptosis images from Figure 2D with the same images from Chen W, Lu Q, Li S, Zhang X, Xue X. microRNA-1298 inhibits the malignant behaviors of breast cancer cells via targeting ADAM9. Bioscience Reports. 2020;40(12):BSR20201215. https://doi.org/10.1042/BSR20201215 and Zhong Y, Lin H, Li Q, Liu C, Zhong L. Downregulation of long non‑coding RNA GACAT1 suppresses proliferation and induces apoptosis of NSCLC cells by sponging microRNA‑422a. International Journal of Molecular Medicine. 2021;47(2):659–667. https://doi.org/10.3892/ijmm.2020.4826. Specifically, Figure 2D, PI and Annexin V-FITC staining cell apoptosis images, have been duplicated with the same images from Figure 2D, PI and Annexin V-FITC staining cell apoptosis images, from Chen et al (2020) and Figure 2E, PI and Annexin V-FITC staining cell apoptosis images, from Zhong et al (2021). The authors responded to our queries but were unable to provide an explanation for the duplicated images nor were they able to provide satisfactory original data from their study. As the findings could not be verified the Editor and Publisher made the decision to retract the article and the authors agreed with this decision. We have been informed in our decision-making by our editorial policies and the COPE guidelines. The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as “Retracted”.

Zhijie Wang,1,2 Yinhua Huang,1,2 Feixue Chu,3 Shangli Ji,2 Kai Liao,1,2 Zekai Cui,2 Jiansu Chen,1,2,4,5 Shibo Tang1,2,6 1Aier School of Ophthalmology, Central South University, Changsha, People’s Republic of China; 2Aier Eye Institute, Aier Eye Hospital Group, Changsha, People’s Republic of China; 3Department of Ophthalmology, Hangzhou Xihu Zhijiang Eye Hospital, Hangzhou, People’s Republic of China; 4Key Laboratory for Regenerative Medicine, Jinan University, Guangzhou, People’s Republic of China; 5Institute of Ophthalmology, Jinan University, Guangzhou, People’s Republic of China; 6CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, People’s Republic of ChinaCorrespondence: Shibo Tang; Jiansu Chen Email tangshibo@vip.163.com; chenjiansu2000@163.comPurpose: Retinal inflammation is involved in the pathogenesis of several retinal diseases. As one of the core clock genes, Nr1d1 has been reported to suppress inflammation in many diseases. We investigated whether pharmacological activation of Nr1d1 can inhibit retinal inflammation and delineated the mechanisms of Nr1d1 in alleviating microglia activation.Methods: Lipopolysaccharide (LPS) induced mice models were used to examine the effects of SR9009 (agonist of NR1D1) treatment on inflammatory phenotypes in vivo. Anti-inflammatory effects of Nr1d1 and associated mechanisms were investigated in the BV2 microglia cell line, and in primary retinal microglia in vitro.Results: SR9009 treatment alleviated LPS-induced inflammatory cell infiltration, elevated cytokine levels and morphological changes of the microglia in mice models. In LPS-stimulated BV2 cells and primary retinal microglia, SR9009 suppressed cytokine expressions by inhibiting the NF-κB signaling pathway. Moreover, SR9009 treatment increased the levels of the M2 phenotype marker (CD206) and the proportions of ramified microglia. Suppression of Nr1d1 with siRNA reversed the inhibitory effects of SR9009 on cytokine production in BV2 cells. RNA-seq analysis showed that genes that were upregulated following Nr1d1 knockdown were enriched in inflammatory-associated biological processes. Subsequently, ChIP-seq of NR1D1 in BV2 was performed, and the results were integrated with RNA-seq results using the Binding and Expression Target Analysis (BETA) tool. Luciferase assays, electrophoretic mobility shift assay (EMSA), qPCR and Western blotting assays revealed that NR1D1 binds the promoter of Hmga2 to suppress its transcription. Notably, overexpressed Hmga2 in activated microglia could partly abolish the anti-inflammatory effects of Nr1d1.Conclusion: The clock gene Nr1d1 protects against retinal inflammation and microglia activation in part by suppressing Hmga2 transcription.Keywords: clock genes, Rev-erbα, Nr1d1, retinal inflammation, Hmga2

Lifang Cheng,1,2,&ast; Kangla Liao,2,&ast; Yingying Wang,2 Fajin Lv,1 Xingming Guo,3 Yineng Zheng,1 Jian Qin2 1Department of Radiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of China; 2Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of China; 3Key Laboratory of Biorheology Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yineng Zheng; Jian Qin Tel/Fax +86-23-89012558Email yinengzheng@cqmu.edu.cn; hjqj2001@163.comBackground: This study aimed to investigate the correlation between the ratio of diastolic to systolic durations (D/S) and echocardiographic parameters of patients with chronic heart failure (CHF) and evaluate whether the D/S can be used as a supplementary biomarker for the classification of heart failure (HF) phenotypes.Methods: In total, 122 CHF patients with a left ventricular ejection fraction (LVEF) < 40%, 40%≤LVEF< 50%, or ≥ 50% were categorized as having HF with a reduced ejection fraction (HFrEF) (N=32), HF with a mid-range ejection fraction (HFmrEF) (N=21) or HF with a preserved ejection fraction (HFpEF) (N=69), respectively. All patients underwent echocardiography for assessment of nineteen structural and functional echocardiographic parameters and digital phonocardiography for the measurement of D/S. Spearman correlation was used to analyse the associations between the D/S and echocardiographic parameters. Multivariate logistic regression analysis was performed to examine the associations between the D/S and HF phenotypes, and receiver operating characteristic (ROC) curve analysis was employed to evaluate the predictive value of the D/S in the classification of HF phenotypes.Results: The D/S values of patients with HFrEF, HFmrEF and HFpEF were 1.32± 0.06, 1.44± 0.11 and 1.54± 0.08, respectively, which were significantly different (All P< 0.05). A close correlation between the D/S and LVEF was found (r=0.777, P< 0.001). The multivariate analysis indicated that the D/S was an independent risk factor for CHF phenotypes (OR=4.927, 95% CI 2.532– 9.587; P< 0.001). The area under the ROC curve for distinguishing between HFmrEF and HFpEF using the D/S was 0.764 (95% CI 0.707– 0.845; P < 0.001) and that for distinguishing between HFmrEF and HFrEF using the D/S was 0.821 (95% CI 0.755– 0.882; P < 0.001).Conclusion: The D/S was significantly associated with LVEF, and as LVEF decreased, the D/S tended to decrease, which could also serve as a noninvasive supplementary indicator for detecting systolic and diastolic dysfunction.Keywords: phonocardiogram, heart sounds, ratio of diastolic to systolic durations, D/S, chronic heart failure, CHF

Mengjie Hu,1,&ast; Songling Chai,2,&ast; 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&ast;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

Wei Zhang,1 Kai Liao,1 Dongning Liu2 1Department of General Surgery, People’s Hospital of Yichun City, Yichun, Jiangxi 336000, People’s Republic of China; 2Department of General Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province 336000, People’s Republic of ChinaCorrespondence: Dongning LiuDepartment of General Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province 330006, People’s Republic of ChinaEmail quedongd22525@126.comBackground: Increasing evidence suggests that microRNAs (miRNAs) play critical roles in cancer progression. Therefore, investigating the function of miRNAs that are aberrantly expressed in gastric cancer (GC) and characterizing the involved underlying mechanism are essential for the treatment of gastric cancer. MiR-138-5p was found to be down-regulated in multiple cancers, which acted as a tumor suppressor in cancer progression; however, whether and how miR-138-5p regulates the malignant behaviors of GC has not been fully understood.Methods: The level of miR-138-5p in GC tissues and cell lines was detected by RT-qPCR. The effects of miR-138-5p on the growth of GC cells were evaluated by the in vitro Cell Counting Kit-8 (CCK-8) assay, cell apoptosis, cell cycle analysis, wound-healing assay, and in vivo xenograft mice model. The targets of miR-138-5p were predicted using the miRDB online tool, confirmed by luciferase report assay and Western blot.Results: MiR-138-5p was frequently decreased in GC tissues and cell lines. Decreased expression of miR-138-5p was significantly associated with the lymph node metastasis of GC patients. Overexpression of miR-138-5p suppressed GC cell proliferation, migration, increased cell apoptosis as well as inhibited the tumor growth in vivo. DEK oncogene was predicted as a potential target of miR-138-5p. MiR-138-5p bound the 3ʹ-UTR of DEK and inhibited the level of DEK in GC cells. Restoration of DEK abrogated miR-138-5p overexpression-mediated suppression of GC cell proliferation and cell cycle arrest.Conclusion: Our results demonstrated the anti-cancer role of miR-138-5p in GC by targeting DEK, which suggested miR-138-5p as a potential therapeutic target for the treatment of patient with GC.Keywords: gastric cancer, miR-138-5p, DEK

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

A preliminary investigation was carried out by the US Environmental Protection Agency (EPA) on the impact of regional ozone concentrations on climate change in the country. The main objective of the investigation was to enhance the ability of air quality managers to consider global change in their decisions through improved characterization of the potential effects of global change on air quality, including O3, PM, and Hg. An integrated assessment framework was designed that supported the research and development efforts within the EPA and the academic research professionals. This design recognized the challenges of bridging spatial scales, temporal scales, and disciplines that characterize the global change and regional air quality problem. A major component of the assessment approach involved the development and application of global to regional climate and air quality modeling systems.