Introduction: The macrolide-resistant Bordetella pertussis (MRBp) has appeared in Asia and has even been prevalent in China. Since the antibiotic sensitivity test is not carried out in the clinical setting, macrolide is still the first choice of antibiotic in MRBp infection. Further, the macrolide therapy for pertussis needs to be revised. Macrolide has always shown a positive effect on other macrolide-resistant bacterium infections in clinical applications. However, the mechanism of macrolide on MRBp remains unclear. Objective: The objective of this study was to investigate the effect of virulence of MRBp under the sub-MIC erythromycin. Methods: This study evaluated a representative isolate BP19147 (ptxP1/fhaB3-MRBp) under a series of sub-inhibitory concentrations of erythromycin. We measured the growth curve, biofilm formation, and autoaggregation assay under Stainer and Scholte (SS) broth. The relative gene expression was detected by RT-qPCR. Results: The proteomics was detected by label-fee DIA. The growth ability and virulence factors of MR isolate BP19147 were inhibited by sub-MIC of erythromycin and had a concentration- dependent effect. From the proteomics results, the pertussis toxin, filamentous haemagglutinin, and pertactin did not show a statistical difference (p >0.05). Other virulence factors (including dermonecrotic toxin, Invasive Adenylate cyclase/haemolysin. etc) showed a statistical difference (p <0.05). In the KEGG enrichment, the BvgAS system, biofilm formation, and some adaptive systems were inhibited by erythromycin. Conclusion: The sub-MIC of erythromycin may reduce the virulence of MRBp, which will provide a theoretical basis for the rational use of erythromycin for MRBp infection and help the development of new antibiotics.

Introduction Asthma is a chronic inflammatory disease characterized by complex immune dysregulation. This study aims to identify key m6A-related genes, particularly METTL3, as potential biomarkers and therapeutic targets in asthma. Methods Transcriptomic data from the GEO (GSE134544) dataset were analyzed to identify differentially expressed m6A-related genes. Functional enrichment analysis was performed using clusterProfiler, immune infiltration profiling was conducted with CIBERSORT, and a competing endogenous RNA (ceRNA, including miRNA and lncRNA) network was constructed. Drug enrichment analysis was carried out using DSigDB, and molecular docking was utilized to assess the interaction between Dabigatran and the METTL3 protein. Results From 192 differentially expressed genes, four m6A-related genes (METTL3, HNRNPC, IGFBP2, and RBMX) were identified as hub genes. Gene Ontology (GO) analysis revealed significant enrichment in biological processes related to RNA metabolic processes and post-transcriptional regulation, while KEGG analysis identified important pathways such as spliceosome and p53 signaling pathways. METTL3 and HNRNPC were central in the ceRNA network, interacting with miRNAs such as hsa-miR-93-3p and lncRNAs like LINC01529. Drug enrichment analysis identified Dabigatran as a potential METTL3 inhibitor, with molecular docking confirming a stable binding affinity (-5.9 kcal/mol). Conclusion This study emphasizes the critical role of m6A-related genes, particularly METTL3 and HNRNPC, as biological macromolecules in asthma pathophysiology, and provides insights into their potential as biomarkers and therapeutic targets for asthma treatment.

The strikingly rapidly mutating nature of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome has been a constant challenge during the coronavirus disease 2019 (COVID-19) pandemic. In this study, various techniques, including reverse transcription-quantitative polymerase chain reaction, antigen-detection rapid diagnostic tests, and high-throughput sequencing were analyzed under different scenarios and spectra for the etiological diagnosis of COVID-19 at the population scale. This study aimed to summarize the latest research progress and provide up-to-date understanding of the methodology used for the evaluation of the immunoprotection conditions against future variants of SARS-CoV-2. Our novel work reviewed the current methods for the evaluation of the immunoprotection status of a specific population (endogenous antibodies) before and after vaccine inoculation (administered with biopharmaceutical antibody products). The present knowledge of the immunoprotection status regarding the COVID-19 complications was also discussed. Knowledge on the immunoprotection status of specific populations can help guide the design of pharmaceutical antibody products, inform practice guidelines, and develop national regulations with respect to the timing of and need for extra rounds of vaccine boosters.

The macrolides-resistant Bordetella pertussis (MR-Bp) isolates in China evolved from the ptxP1/fhaB3 allele and rapidly became predominant, suggestive of an adaptive transmission ability. This was different from the global prevalent ptxP3 strains, in which MR-Bp was rarely reported. The study aimed to determine the underlying mechanism responsible for fitness and resistance in these two strains. We identify proteomic differences between ptxP1/fhaB3 and ptxP3/fhaB1 strains using tandem mass tag (TMT)-based proteomics. We then performed in-depth bioinformatic analysis to determine differentially expressed genes (DEGs), followed by gene ontology (GO), and protein-protein interaction (PPI) network analysis. Further parallel reaction monitoring (PRM) analysis confirmed the expression of four target proteins. Finally, the crystal violet method was used to determine biofilm-forming ability. The results showed that the main significantly different proteins between the two represent isolates were related to biofilm formation. Furthermore, we have confirmed that ptxP1/fhaB3 showed hyperbiofilm formation in comparison with ptxP3/fhaB1. It is suggested that the resistance and adaptability of ptxP1/fhaB3 strains may be related to the formation of biofilm through proteomics. In a word, we determined the significantly different proteins between the ptxP1/fhaB3 and ptxP3/fhaB1 strains through whole-cell proteome, which were related to biofilm formation.

Background: Microbiome is an important internal ecosystem closely related to host health. Most of the bacteria existed in the internal ecosystem cannot be isolated with laboratory bacteriological culture methods, while 16S rDNA sequencing is considered and used for the bacterial identification by through the high-throughput platforms. The aim of this study was to compare the microbiota analysis result using two next-generation sequencing platforms and bioinformatics pipelines. Results: 56 maternal-neonate fecal samples were sequenced and analyzed by 16S rRNA amplicon sequencing both by Ion Torrent S5-xl and Illumina Hiseq 2500 with standard protocols at same lab. For the richness and diversity of microbiota, index of chao1, observed_specise, PD_whole_tree, simpson and good_converage varied significantly except Shannon index at two platforms (P<0.05). The relative abundance of bacteria at different taxonomy levels is checked from phylum to species level, the more species of bacteria sequenced and annotated, the lower the correlation of the relative abundance of the bacteria founded between two platforms. The sequencing results are consistent between two platforms. Principal component analysis (PCA) results showed that more than 87% of samples were concentrated. According to principal coordinate analysis (PCoA), 56 samples of the two platforms were divided into two clusters, and the compliance rate of the two platforms is 71.43%. The differences between microbial community structures generated from two platforms were tested by multi-response permutation procedure (MRPP), which showed significant differences at family and genus levels separately (A=0.094, P=0.001; A=0.085, P=0.002). When maternal and neonate samples were considered, at family level, there was no difference in microbiota composition between two platform for maternal group (A=0.006, P=0.149), while in the neonate group, it showed significant differences (A=0.035, P=0.006). At the genus level, there existed significant differences in microbiota both in maternal and neonate group (A=0.0216, P=0.004; A=0.098, P=0.001). Conclusion: Although the relative abundance of microbiota sequenced at two different platforms is basically similar, the diversity and correlation coefficient are still quite different. To increase reproducibility and reliability in cohort studies, it is important to use the same sequencing platforms and the corresponding pipeline to reduce the systematic error in microbiome analysis.

Probiotics may offer an attractive alternative for standard antibiotic therapy to treat Clostridium difficile infections (CDI). In this study, the antibacterial mechanism in vitro of newly isolated B. amyloliquefaciens C-1 against C. difficile was investigated. The lipopeptides surfactin, iturin, and fengycin produced by C-1 strongly inhibited C. difficile growth and viability. Systematic research of the bacteriostatic mechanism showed that the C-1 lipopeptides damage the integrity of the C. difficile cell wall and cell membrane. In addition, the lipopeptide binds to C. difficile genomic DNA, leading to cell death. Genome resequencing revealed many important antimicrobial compound-encoding clusters, including six nonribosomal peptides (surfactins (srfABCD), iturins (ituABCD), fengycins (fenABCDE), bacillibactin (bmyABC), teichuronic, and bacilysin) and three polyketides (bacillaene (baeEDLMNJRS), difficidin (difABCDEFGHIJ), and macrolactin (mlnABCDEFGHI)). In addition, there were other beneficial genes, such as phospholipase and seven siderophore biosynthesis gene clusters, which may contribute synergistically to the antibacterial activity of B . amyloliquefaciens C-1. We suggest that proper application of antimicrobial peptides may be effective in C . difficile control.

目的 调查西安地区泌尿道感染老年患者的病原菌分布及耐药特点.方法 回顾性分析2012年11月至2017年3月在西安市某三甲医院就诊的2475例65岁及以上泌尿道感染患者的尿标本病原菌信息及其药敏试验数据.结果 从1465例女性标本和1010例男性标本中共分离出2475株非重复病原菌,其中革兰阴性杆菌1594株(64.40％),以大肠埃希菌为主,为1230株(49.70％);革兰阳性球菌452株(18.26％),以屎肠球菌为主,为294株(11.88％).大肠埃希菌是导致老年女性泌尿道感染的主要病原菌,其感染率(61.98％)明显高于男性(31.88％),差异有统计学意义(χ2=216.643,P<0.001).病原菌对抗菌药物的耐药率:氨苄西林为82.76％,环丙沙星为69.68％,左氧氟沙星为65.52％,呋喃妥因为20.33％.女性分离株对氨苄西林的耐药率(84.08％)高于男性(80.45％),差异有统计学意义(χ2=4.210,P=0.040);男性分离株对呋喃妥因的耐药率(28.07％)明显高于女性(15.92％),差异有统计学意义(χ2=41.550,P<0.001).女性产超广谱β-内酰胺酶的大肠埃希菌的耐药率(52.75％)低于男性(79.19％),差异有统计学意义(χ2=69.051,P<0.001);女性耐碳青霉烯类肠杆菌科细菌的耐药率(0.22％)低于男性(0.62％),但差异无统计学意义(χ2=1.178,P=0.278).结论 该地区老年泌尿道感染患者以女性为主,病原菌除大肠埃希菌外,应重视对抗菌药物耐药程度较高的屎肠球菌.老年男性泌尿道感染患者,应避免过量使用氨苄西林 、环丙沙星和左氧氟沙星进行经验性治疗.

目的 建立粪便中乙酸、丙酸、正丁酸、异丁酸、正戊酸和异戊酸等六种短链脂肪酸(SCFAs)的快速提取及气相色谱定量检测方法.方法 粪便样本分别采用纯水和盐酸溶液(pH=2)提取,静置、离心后,上清液中的SCFAs采用气相色谱法测定.以正丁醇为内标,保留时间定性,峰面积定量.结果 本法线性关系良好,水提取和酸提取的加标回收率分别为78.8％～ 115％和85.7％～130％,且无统计学差异(P＞0.05).比较人和鼠的粪便水提法所得SCFAs含量与参考文献检测结果吻合.结论 本法高效、稳定,可应用于大批粪便样本6种短链脂肪酸的快速测定.

The aim of this study was to evaluate the probiotic characteristics and safety of seven Enterococcus faecalis isolates from fecal samples of healthy Chinese infants. We evaluated the isolates' tolerance to low pH, survival in bile salts and NaCl, adhesion ability, biofilm formation, antimicrobial activity, toxin gene distribution, hemolysis, gelatinase activity, antibiotic resistance, and virulence to Galleria mellonella. All strains survived at pH 5.0, in 7.0% NaCl, and in 3% bile salt. Adhesion to Caco-2 cells was above 10%. Strain A3-1 had higher adhesion ability toward mucin, collagen, and BSA in vitro, better antibacterial activity, and the strongest biofilm production. We detected seven virulence genes with a distribution of asa1 (100%), cylA (71.4%), esp (85.7%), hyl (14.3%), gelE (85.7%), ace (42.9%), and agg (71.4%). Although all strains were γ-hemolytic, none showed gelatinase activity based on physiological activity detection. All isolates were susceptible to benzylpenicillin, ampicillin, ciprofloxacin, levofloxacin, moxifloxacin, tigecycline, nitrofurantoin, linezolid, and vancomycin; they were not susceptible to erythromycin, quinupristin/dalofopine, and clindamycin. The virulence test of G. mellonella showed that, except for strains 106-1 and 113-1, the other strains had toxicity lower than 10%. Strain A3-1 may have the greatest potential to be developed as a probiotic.