Modified Ganlu Xiaodu decoction attenuates airway barrier injury in Mycoplasma pneumoniae pneumonia by inhibiting ZBP1-mediated PANoptosis.

Comparison of lower and upper respiratory tract specimens for the detection of respiratory viruses and atypical pneumonia pathogens by multiplex real-time polymerase chain reaction.

Respiratory pathogens jeopardize population health, particularly high-risk groups. CRISPR-Cas systems, as novel nucleic acid detection platforms, offer timely identification and have become a major research focus. This study presents a novel diagnostic workflow that combines recombinase polymerase amplification (RPA) for pre-amplification of pathogen nucleic acids with CRISPR-based detection. By combining microfluidic technology and portable imaging devices, this study developed a multiplex assay capable of simultaneously detecting seven clinically relevant pathogens in a single sample, including influenza A virus (FluA), influenza B virus (FluB), respiratory syncytial virus (HRSV) A and B, mycoplasma pneumoniae (MP), adenovirus (HAdv), and parainfluenza virus (HPIVs). Utilizing the POCT-CRISPR platform, simultaneous detection of seven respiratory pathogens can be achieved within approximately 30 min, achieving detection limits of 0.1-1 fM. This method streamlines the detection process, significantly reducing both the complexity of operations and the overall detection time. Clinical cohort validation demonstrated a detection efficiency of 99.63% sensitivity and 100% specificity. These results confirm the effectiveness and reliability of the detection method. Additionally, the 7-virus panel is estimated at approximately $32 per sample, a cost competitive with commercial multiplex qPCR detection kits ($15-$110 per sample) and substantially more economical than integrated cartridge-based syndromic platforms. The platform features simple operation, cost-effectiveness, short turnaround time, and reliable detection performance, making it highly suitable for point-of-care testing (POCT) at the grassroots level.

PurposeWe analyzed the epidemiological and etiological characteristics of six prevalent pathogens responsible for respiratory infections in pediatric patients.Patients and MethodsThis retrospective cross-sectional study involved 6134 children with respiratory infections from January 2024 to December 2024. Nasopharyngeal swabs were collected and analyzed using multiplex PCR to detect Influenza A (IFVA) and B viruses (IFVB), Respiratory Syncytial Virus (RSV), Adenovirus (ADV), Human Rhinovirus (HRV), and Mycoplasma pneumoniae (MP). Pathogen distribution was assessed.ResultsAmong the six pathogens tested, HRV and ADV were the most frequently detected, followed by MP, RSV, IFVB, and IFVA. Single-pathogen infections accounted for 62.3% of cases, while co-infections of two, three, and four pathogens comprised 13.5%, 2.2%, and 0.08%, respectively. The most common co-infection pattern was HRV+ADV. Age-specific analysis revealed distinct shifts: RSV predominated in infants under 1 year, HRV and ADV were most common in preschool children aged 3–<7 years, and MP dominated in school-aged children aged 7–14 years (all P < 0.001 across age groups). Seasonally, RSV peaked in spring and winter, ADV in summer, HRV in winter, and IFVB showed an unusual spring peak (all P < 0.001). No significant sex differences were observed except for ADV, which was slightly higher in males (P = 0.009).ConclusionThis first comprehensive analysis of pediatric respiratory infections in Xinjiang reveals epidemiological patterns distinct from other Chinese regions, characterized by atypical seasonal peaks (spring IFVB, summer ADV, winter HRV) and age-specific pathogen shifts (RSV in infants, MP in school-aged children), with HRV+ADV as the dominant co-infection. These findings highlight the necessity of region-tailored prevention strategies and provide a baseline for future surveillance in understudied continental climate populations.

This study aimed to assess macrolide resistance and the genotype distribution of Mycoplasma pneumoniae (MP) in children and identify the risk factors for refractory MP pneumonia (RMPP) during the initial post-COVID-19 outbreak in Wuhu. This retrospective study enrolled children with MPP in Wuhu (November 2023–May 2024). Using preserved nasopharyngeal swabs, macrolide resistance mutations were identified by sequencing the 23S rRNA domain V, and genotyping was performed by PCR. Clinical and laboratory parameters were analyzed to identify the risk factors associated with RMPP. An analysis of 296 pediatric MPP patients revealed universal cough (100%), with high rates of fever (94.93%) and sputum (71.62%). Common findings included moist rales (53.04%) and lobar pneumonia (37.5%). Genotype I (87.84%) predominated and showed a higher A2063G resistance mutation rate than genotype II (74.23% vs 55.56%, P = .019). This mutation was associated with longer hospitalization, more vomiting, and higher C-reactive protein (CRP). RMPP (69.26%) was notable for producing more severe symptoms, higher inflammatory markers (CRP, lactate dehydrogenase [LDH], procalcitonin [PCT], D-D dimer), and a greater need for bronchoscopy and steroid use. Multivariate logistic regression identified prolonged hospitalization, the presence of pulmonary infiltrates, CRP > 10 mg/L, LDH > 246 U/L, and D-D dimer > 0.50 μg/mL as independent risk factors for RMPP. Subsequent receiver operating characteristic curve analysis demonstrated the predictive utility of D-D dimer and LDH levels. The present study established genotype I MP with A2063G-mediated macrolide resistance as the predominant epidemic variant in Wuhu. Independent risk factors for RMPP included a prolonged hospital stay and elevated CRP, LDH and D-D dimer. LDH combined with D-D dimer served as a valuable co-marker for early risk stratification and prediction of refractory disease.

Background:This full-scale confirmatory study aimed to measure serum neutrophil extracellular traps (NETs) levels in children with severe Mycoplasma pneumoniae pneumonia (SMPP) and non-severe MPP, and to compare the predictive performance of serum NETs, C-reactive protein (CRP), and lactate dehydrogenase (LDH) for SMPP.Methods:A total of 200 children hospitalized with MPP between January 2023 and August 2024 were consecutively enrolled and divided into SMPP (n = 100) and non-severe MPP (n = 100) groups. Baseline demographic and laboratory indices were compared between groups. Receiver operating characteristic (ROC) curves were used to assess predictive value, and univariate and multivariate logistic regression were performed to identify independent predictors of SMPP.Results:Age and sex distributions were comparable between groups (all P > .05). Serum white blood cell count, neutrophil ratio, CRP, D-dimer, LDH, and NETs were significantly higher in the SMPP group (all P < .001). ROC analysis showed serum NETs had the highest area under the curve (AUC = 0.83, 95% confidence interval [CI]: 0.762–0.896), with a cutoff value of 19.2, sensitivity 0.818, and specificity 0.835. Multivariate logistic regression confirmed serum NETs (odds ratio [OR] = 4.32, 95% CI: 3.45–7.25, P = .023), CRP (OR = 3.62, 95% CI: 1.85–9.25, P = .010), and LDH (OR = 1.63, 95% CI: 1.18–2.26, P = .001) as independent predictors of SMPP.Conclusion:Serum NETs are strongly elevated in children with SMPP and show better predictive performance than conventional CRP and LDH. Combined measurement of NETs, CRP, and LDH provides a reliable tool for the early identification of pediatric SMPP. This study extends and validates our preliminary exploratory findings with a larger sample and complete statistical analysis.

To characterize co-infections in children with respiratory syncytial virus (RSV) and evaluate their association with disease severity. We retrospectively analyzed 5, 586 children hospitalized with RSV infection at Tianjin Children's Hospital between January 2018 and December 2023. Patients were grouped by co-infection status: RSV alone, RSV with viral co-infection, RSV with bacterial co-infection, RSV with Mycoplasma pneumoniae (MP) co-infection, and RSV with multiple co-infections. Demographic characteristics, clinical features, and disease severity were compared across groups. Of the 5, 586 patients, 3, 835 had RSV alone, 476 had viral co-infections, 715 had bacterial co-infections, 312 had MP co-infections, and 248 had multiple co-infections. Children with multiple co-infections were older (median 3 vs. 0.5 years; P < 0.05), had longer fever and illness duration, and showed higher rates of severe pneumonia (all P < 0.05). Viral co-infections were more often associated with wheezing and chest retractions, while bacterial and MP co-infections were characterized by fever and cough. Co-infections in RSV-infected children were associated with distinct clinical patterns and differences in disease severity. Multiple co-infections were associated with a higher proportion of severe pneumonia and prolonged illness. Identifying co-infection patterns may help detect high-risk patients early and guide individualized management. Not applicable.

Increased detection rate of macrolide-resistant Mycoplasma pneumoniae during the Mycoplasma pneumoniae epidemic in children: A multi-campus hospital retrospective study in Guangzhou, 2023-2024.

ABSTRACT Mycoplasma pneumonia (MP), as a global infectious disease in sheep, seriously affects the production performance of sheep and economic benefits of sheep industry. However, current research on sheep resistance to MP remains limited. To address this gap and explore the potential epigenetic regulation of sheep MP resistant, this study employed high-throughput sequencing techniques (ATAC-Seq and CUT&Tag) to analyze epigenetic modifications in lung tissue from healthy and MP-affected sheep and reveal differential epigenetic landscapes associated with disease resistance. Integrating transcriptome analysis related to MP and genome-wide association studies (GWAS), FOXF1 was identified as a candidate gene for MP-resistance in sheep. We established a Mycoplasma ovipneumoniae (MO)-infected sheep alveolar epithelial cell model and regulated FOXF1 expression in cells through interference and overexpression techniques to study MO’s adhesion and damage. The results showed that activation promoters or enhancer elements in FOXF1 introns of healthy lungs may enhance its transcription. FOXF1 overexpression reduced MO-mediated adhesion damage to cells, while knock-down increased it. Our work has enriched the gene pool for anti-pneumonia and studied the role of the FOXF1 gene in MO-infected cells, accumulating reliable genetic resources for sheep MP disease resistant breeding.

Background/Objectives: Mycoplasma pneumoniae (MP) is a frequent cause of community-acquired pneumonia, but it is increasingly recognized for extrapulmonary complications, specifically Mycoplasma pneumoniae-induced rash and mucositis (MIRM). This systematic review aims to comprehensively assess the frequency of clinical features, diagnostic criteria and outcomes of oral mucositis in patients with confirmed MP infection. Methods: A systematic review was conducted following PRISMA guidelines across PubMed, Web of Science and Scopus, covering the period 2015–2025. Inclusion criteria encompassed in vivo studies, case reports, and case series in English focusing on MP-associated mucositis. Methodological quality was assessed using JBI checklists for case-based evidence and the Newcastle–Ottawa Scale for cohort studies. Two clinical cases were reported. Results: Out of 242 identified records, 42 studies were included, involving 140 patients with a notable male predominance (62%). Oral involvement was reported in 92.9% of cases, often characterized by severe ulcerations, hemorrhagic crusting, and debilitating pain. Intensive Care Unit admission was required in 21.5% of cases due to severe systemic or mucosal disease, with 14.3% necessitating parenteral nutrition. Quality assessment indicated moderate-to-high methodological rigor across most included studies. Conclusions: MIRM represents a significant clinical entity where oral mucositis is a dominant feature, often preceding or overshadowing respiratory symptoms. Early recognition by oral health professionals is crucial to avoid misdiagnosis, ensure appropriate multidisciplinary care, and implement supportive or immunomodulatory therapies that reduce morbidity and hospitalization length.

Objective: To explore the differences in clinical characteristics, laboratory indicators, and outcomes of Mycoplasma pneumoniae (MP) necrotizing pneumonia (NP) in children with co-infection of bacteria or viruses. Methods: A retrospective cohort study was conducted. Clinical data of 109 children with MPNP hospitalized in the Department of Infectious Diseases and Pediatric Intensive Care Unit of the Children's Hospital, Zhejiang University School of Medicine from January 2019 to March 2024 were included. Based on pathogen results, the children were divided into a single MP infection group, an MP co-infection with bacteria group, and an MP co-infection with viruses group. Demographic data, clinical manifestations, laboratory indicators, bronchoscopic findings, and clinical outcomes were compared among the three groups. Inter-group comparisons were performed using independent samples t-test, Kruskal-Wallis H test, Chi-square test, and Fisher's exact test. Length of hospital stay among the three groups was analyzed using Kaplan-Meier curves. Results: Among the 109 MPNP children, there were 50 males and 59 females, with an onset age of 7.1 (5.5, 8.3) years. There were 68 cases in the single MP infection group, 30 cases in the MP co-infection with viruses group, and 11 cases in the MP co-infection with bacteria group. Statistically significant differences were observed among the three groups in terms of post-admission fever duration, rate of closed drainage of thoracic cavity, and serum interleukin (IL)-6 levels (all P<0.05). Among the 65 children who underwent a second bronchoalveolar lavage, there was a statistically significant difference in the incidence of persistent mucus plugs among the three groups (42.9% (18/43) vs. 12/15 vs. 3/7, P<0.05). Conclusions: In children with MPNP co-infected with bacteria, clinicians should be aware of the possibility of longer fever duration, higher rates of thoracic drainage, and higher levels of IL-6. Co-infection with viruses is associated with a higher proportion of persistent mucus plugs, which may necessitate more aggressive bronchoscopic intervention.

Verification of the JRS adult pneumonia clinical practice guidelines 2024: Sensitivity and specificity of the mycoplasma pneumonia diagnostic prediction score.

To explore the risk factors and predictive value of Mycoplasma pneumoniae (MP) infection in children. A total of 2042 children with suspected Mycoplasma pneumoniae infection who were treated for the first time at Civil Aviation General Hospital from October 2023 to December 2023 were selected as the study subjects. Among them, 1637 cases were confirmed as Mycoplasma pneumonia-infected and were included in the pneumonia group, while the remaining 405 cases were non-Mycoplasma pneumonia-infected and were included in the non-Mycoplasma pneumonia group. The clinical data of the 2 groups of children (including gender, age, initial symptoms, laboratory indicators, etc) and the risk factors of MP infection in children were compared, and the receiver operating characteristic curve was analyzed. This study showed that the percentage of neutrophils in the non-MP infection group was significantly lower than that in the MP infection group, and the difference was statistically significant (P < .001). When comparing the percentages of lymphocyte percentage (LY) and hemoglobin in the 2 groups of children, the Mycoplasma pneumonia-infected group was lower than the non-Mycoplasma pneumonia-infected group, and the differences were both statistically significant (P < .05). Logistic regression analysis revealed that white blood cell and neutrophil-to-lymphocyte ratio (NLR) might be valuable markers for predicting MP infection. The Spearman correlation indicated that LY was collinear with the occurrence of MP infection, and Least Absolute Shrinkage and Selection Operator (LASSO) regression analysis demonstrated that both LY and NLR might be valuable markers for predicting MP infection (P < .05). Receiver operating characteristic curve analysis revealed that the area under the curve of the NLR for diagnosing MP infection was 0.624, with a cutoff value of 1.36 (sensitivity of 0.798 and specificity of 0.558). In the diagnosis of MP infection, the consistency between the RNA method and the immunogold colloidal method was poor (Kappa = 0.108, P < .05). The consistency between the RNA method and the immunogold colloidal method in the diagnosis of MP infection is poor. Both the white blood cell and NLR are valuable markers for MP infection.

Background. Mycoplasma pneumoniae (MP) is a common causative pathogen of community-acquired pneumonia in children, with clinical presentations ranging in severity. Early stratification and timely intervention are essential for improving patient outcomes. However, a major clinical challenge lies in the limited ability to accurately distinguish between mild and severe cases based solely on early clinical indicators.Methods. This prospective real-world study investigated the differences in microbiome and metabolomics between mild and severe MP pneumonia (MPP) in children. Bronchoalveolar lavage fluid samples were collected from 153 children and subjected to metagenomic sequencing and non-targeted metabolomic analysis. Meanwhile, to enhance early diagnostic accuracy, this study developed a machine learning classification model and validated it using a third-party validation set.Results. The results revealed significant alterations in the abundance of specific bacterial communities in the severe group, most notably the coexistence of MP and Alphainfluenzavirus influenzae, which may contribute to disease exacerbation through synergistic pathogenic mechanisms. Furthermore, the macrolide resistant rate of MP in the severe group exceeded 80%, emphasizing the importance of appropriate antibiotic selection. Metabolomic analysis showed a significant enrichment of metabolites related to cellular energy metabolism and immune regulation in severe cases. The model demonstrated exceptional predictive performance, achieving an area under the curve ranging from 0.909 to 0.991, which significantly outperformed conventional clinical stratification methods.Conclusions. These findings elucidate the distinct pathophysiological mechanisms underlying both mild and severe MP infections and provide a promising framework for improving early diagnosis and personalized treatment strategies in paediatric MPP.

To compare clinical and imaging features in children with macrolide-susceptible (MSMP) and macrolide-resistant Mycoplasma pneumoniae pneumonia (MRMP) in a real-world clinical setting. We retrospectively analysed paediatric M. pneumoniae (MP) cases at Hunan People's Hospital from June to December 2023, utilizing polymerase chain reaction (PCR) to categorize infections into MRMP and MSMP groups based on 23S rRNA mutations. We compared symptoms, laboratory findings, imaging results and treatment outcomes between MRMP and MSMP groups. Additionally, univariate and multivariate logistic regression analyses were performed to identify factors associated with the switch to doxycycline in MRMP patients. Of 199 children with MP, 151 were categorized as MRMP due to 23S rRNA mutations, while 48 were classified as MSMP. The MRMP group included more preschool- and school-aged children, with more severe symptoms (persistent high fever and extensive lung consolidation). MRMP patients received longer antibiotic courses, with imaging showing segmental or lobar infiltrates. Consequently, treatment was switched from azithromycin to doxycycline in 66.2% of MRMP patients. Multivariate logistic regression analysis revealed that segmental or lobar parenchymal infiltration was independently associated with an increased likelihood of receiving doxycycline (OR = 3.942, p < 0.001). These real-world data suggest that MRMP may present with more severe clinical manifestations and radiological features than MSMP among hospitalized children during an atypical post-COVID resurgence of MP. Doxycycline could be considered as the second-line therapy for MRMP patients exhibiting severe symptoms or lung consolidation when macrolides fail to achieve clinical improvement.

Pneumonia, a respiratory infection that primarily affects the lungs, is a significant health concern in the pediatric population, often leading to hospitalization and, in severe cases, mortality. The present study was performed to investigate the therapeutic effects of the flavokawain A against mycoplasma pneumonia in mice. BALB/c mice were subjected to an exposure with 100 µL of Mycoplasma pneumoniae (Mp) via nasal drips for a duration of 2 days, and subsequently administered 50 mg/kg of flavokawain A for a duration of 3 days. The pulmonary index (PI), survival percentage, and survival time was assessed in the experimental mice. The inflammatory biomarker concentrations (MPO and NO), oxidative stress biomarkers (MDA, SOD, and GSH), C-reactive protein (CRP), Mp-specific Ig-M, and inflammatory cytokines were analyzed with the appropriate assay kits. The Mp-DNA concentration were assessed in the experimental mice. The concentrations of JNK, ERK, and NF-κB in the lung tissues were assessed using kits. The histopathological examination was conducted on lung tissues to evaluate the histological alterations. The flavokawain A treatment significantly reduced the lethality and increased survival time in Mp-induced mice. It also diminished the PI, NO, and MPO concentrations in Mp-induced mice. The flavokawain A administration resulted in a reduction of MDA levels and an enhancement of antioxidants SOD and GSH levels. Flavokawain A treatment significantly diminished the levels of inflammatory cytokines, CRP, Mp-specific Ig-M, and Mp-DNA content in the Mp-infected mice. The histological results demonstrated a significant reduction in inflammatory signs and alveolar injury in the Mp-induced mice. The current findings confirm the salutary properties of flavokawain A against Mp-infected mice. Consequently, flavokawain A may serve as a talented therapeutic agent to treat pneumonia.

Rapid and accurate detection of respiratory pathogens using a field-deployable POCT system based on multiplex real-time PCR.

Enhancing effects of Mycoplasma pneumoniae antigens on Th2-attracting chemokine responses by murine splenocytes.

BackgroundRefractory and severe evolution complicate pediatric Mycoplasma pneumoniae pneumonia (MPP), yet early risk stratification still relies largely on single-time admission biomarkers. We tested whether a prespecified 48-hour pre-admission fever-burden index (FBI48) predicts in-hospital outcomes and improves model performance beyond guideline-consistent clinical and laboratory predictors.MethodsWe conducted a retrospective single-center cohort study of hospitalized children ≤14 years with laboratory-confirmed MPP. FBI48 was defined as the area under the temperature–time curve above 38.0 °C over −48 to 0 h (°C·h). Thresholds of 38.5 °C and 39.0 °C were evaluated in sensitivity analyses. Prespecified covariates, chosen based on prior RMPP/SMPP studies and pediatric CAP/MPP guidelines, included age, illness days, SpO2, imaging severity, prior macrolide exposure, antipyretic and steroid use, LDH, log2-transformed NLR, CRP, and PCT. Penalized logistic regression generated predicted risks, while unpenalized models provided adjusted odds ratios. Model performance (AUC, Brier score, calibration) and decision-curve analysis (DCA) net benefit were assessed for base (clinical + laboratory) and augmented (base + FBI48) models across 10–30% risk thresholds.ResultsOf 720 eligible hospitalizations, 648 were analyzed. The composite endpoint (RMPP and/or incident SMPP) occurred in 176/648 (27.2%; RMPP 24.7%; SMPP 8.0%). FBI48 was independently associated with the composite endpoint (adjusted odds ratio [aOR] 1.45, 95% CI 1.25–1.68 per 1 SD ≈22 °C·h) and with SMPP alone (aOR 1.58, 95% CI 1.21–2.06). Adding FBI48 to the base model improved AUC from 0.78 to 0.83 (ΔAUC 0.05, p<0.001), reduced the Brier score (0.176 to 0.164, p=0.006), and increased net benefit compared with the base model and a treat-none strategy across 10–30% thresholds. Alternative fever thresholds (38.5 °C/39.0 °C) yielded similar effect sizes.ConclusionsA simple 48-hour pre-admission fever-burden metric provides independent and incremental prognostic information on the risk of refractory or severe evolution in pediatric MPP, complementing guideline-based clinical and laboratory predictors and supporting admission-time risk stratification. External validation and prospective evaluation of dynamic, in-hospital updating are warranted.

In recent years, Mycoplasma pneumoniae (MP) has developed widespread resistance to macrolide antibiotics, and second-line anti-Mycoplasma drugs such as levofloxacin have begun to be used in children. However, there is currently insufficient data on their efficacy and safety. To explore the application effect of levofloxacin in children with refractory Mycoplasma pneumoniae pneumonia (RMPP) and evaluate its safety. This study is an exploratory research, selecting 82 RMPP patients treated with levofloxacin who were admitted to the Department of Pediatric Respiratory Medicine at Weifang People's Hospital between November 2023 and December 2024 as the research subjects. Compared the blood routine and biochemical indicators before and after the application of levofloxacin, and observed the clinical efficacy and adverse reactions of levofloxacin. The average fever reduction time after the application of levofloxacin was (1.96 ± 1.52) days. After the application of levofloxacin, there was no change in white blood cell (WBC), neutrophil (N), platelet (PLT) and other parameters in the blood routine (P > 0.05). The lymphocyte count (L) has increased but remains within the normal range. There was no difference in alanine aminotransferase (ALT) before and after treatment, but aspartate aminotransferase (AST) decreased compared to before. Lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CKMB) decreased after medication (P < 0.05), while creatine kinase (CK) remained unchanged. Blood creatinine (CREA) and Blood urea (Urea) both decreased (P < 0.05), indicating no damage to liver and kidney function. There were a total of 9 adverse drug reactions related to levofloxacin, including 5 cases of rash, 2 cases of gastrointestinal reactions, 1 case of neurological reaction, and 1 case of lower limb pain. No other adverse reactions or sequelae were observed during the 12-month clinical follow-up. Levofloxacin has shown good efficacy in treating RMPP in children, with no serious adverse reactions detected in a short period of time.