Frontiers in Cellular and Infection Microbiology

Introduction Typhoid fever is an infectious disease primarily caused by Salmonella enterica sv. Typhi ( S. Typhi), a bacterium that causes as many as 20 million infections and 600,000 deaths annually. Asymptomatic chronic carriers of S. Typhi play a major role in the transmission of typhoid fever, as they intermittently shed the bacteria and can unknowingly infect humans in close proximity. An estimated 90% of chronic carriers have gallstones; biofilm formation on gallstones is a primary factor in the establishment and maintenance of gallbladder carriage. CsgD is a central biofilm regulator in Salmonella , but the S. Typhi csgD gene has a mutation that introduces an early stop codon, resulting in a protein truncated by 8 amino acids at the C-terminus. In this study, we investigate the role of role of CsgD in S. Typhi. Methods We introduced a fully functional copy of the csgD gene from S. Typhimurium into S. Typhi under both a native and a constitutive promoter and tested for red, dry, and rough (Rdar) colony morphology, curli fimbriae, cellulose, and biofilm formation. Results and discussion We demonstrate that although CsgD-regulated curli and cellulose production were partially restored, the introduction of the S. Typhimurium csgD did not induce the Rdar colony morphology. Interestingly, we show that CsgD does not have a significant role in S. Typhi biofilm formation, as biofilm-forming capacities depend more on the isolate than the CsgD regulator. This data suggests the presence of an alternative biofilm regulatory process in this human-restricted pathogen.

Apiospora is an important genus in the Apiosporaceae family with a worldwide distribution. They exhibit different lifestyles including pathogenic, saprophytic, and endophytic. In this study, we aimed to explore the Apiospora associated with bamboo and collected 14 apiospora-like taxa from the forests of Yunnan Province, China. Morphological and phylogenetic analyses (combined ITS, LSU, tef 1- α , and tub 2 sequence data) confirmed that these collections belong to Apiospora s. str. and reports five new species (viz., Ap. dehongensis, Ap. jinghongensis, Ap. shangrilaensis, Ap. zhaotongensis , and Ap. zhenxiongensis ). New sexual morphs of asexually typified Ap. globose and Ap. guangdongensis species, and a new geographical record of Ap. subglobosa are also reported. The findings of this study not only enhance the diversity of bambusicolous fungi in the region of Yunnan, but also geographical distribution of some known Apiospora species.

Among the diverse fungal opportunistic pathogens, Candida albicans garners significant attention due to its wide range of infections and high frequency of occurrence. The emergence of resistance and the limited number of antifungals drives the need to develop novel antifungal drugs. Although the natural product baicalin has been shown to trigger apoptosis in C. albicans in previous experiments, its influence on cell wall (CW) structure along with immune recognition remains elusive. In this work, baicalin showed a significant killing effect against C. albicans SC5314. Moreover, CW destruction, characterized by β-1,3-glucan unmasking and chitin deposition, was observed as a consequence of the treatment with baicalin. The RNA sequencing analysis revealed that treatment with baicalin resulted in eight hundred forty-two differentially expressed genes (DEGs). Sixty-five genes, such as GSC1, ENG1, CHS3, GWT1, and MKC1, were associated with CW organization or biogenesis. Baicalin-pretreated C. albicans SC5314 was phagocytosed more efficiently by RAW264.7 macrophages, accompanied by increased TNF-α and IL-1β production. Accordingly, it is hypothesized that baicalin could stimulate β-1,3-glucan unmasking by governing CW-associated gene expression in C. albicans SC5314, which contributes to macrophage recognition and clearance.

Introduction This study, conducted in China prior to RotaTeq’s launch, examined the epidemiological, molecular, and evolutionary features of the G1P[8] genotype RVA in children admitted with diarrhea, to aid in evaluating its efficacy and impact on G1P[8] RVA in China. Methods Data from the Chinese viral diarrhea surveillance network were collected from January 2016 to December 2018. RVA strains identified as the G1P[8] genotype were subjected to whole-genome sequencing. Neutralizing epitope, amino acid selection pressure, and evolution dynamics analyses on VP7 and VP4 were performed using BioEdit v.7.0.9.0 and PyMOL v.2.5.2, four algorithms (MEME, SLAC, FEL, and FUBAR) in the Datamonkey online software, and the MCMC model in BEAST v. 1.10.4, respectively. Phylogenetic and identity features of 11 genes were assessed by DNAStar and MEGA v.7. Results Results showed that the detection rate of G1P[8] in China from 2016 to 2018 was generally low with significant seasonality. The whole genome of G1P[8] of four 2016 childhood diarrhea specimens was successfully sequenced. Phylogenetic and neutralizing epitope analysis showed that Rotavin-M1 might have better protection on G1P[8] prevalent in China than Rotarix and RotaTeq. Two conserved N-glycosylation sites on VP7 of Chinese G1P[8] might affect the protective effect of the vaccine. Evolution rate and selection pressure analysis identified the possibility of rapidly evolving and adapting to the new environment introduced by vaccines of G1P[8], whereas positive selection specific to VP4 indicated the potential tendency to select for dominant traits. Identity and phylogeny analysis showed that Chinese G1P[8] from before 2018 was generally stable with possible genetic recombination among local strains. Discussion These findings not only are of great significance for predicting the prevalence of G1P [8] in China, but also provide data reference for evaluating rotavirus vaccine efficacy.

Objectives To investigate the effect of gut microbiota from postmenopausal osteoporosis patients on bone mass in mice. Methods Fecal samples were collected from postmenopausal women with normal bone mass (Con, n=5) and postmenopausal women with osteoporosis (Op, n=5). Microbial composition was identified by shallow shotgun sequencing. Then fecal samples were transplanted into pseudo-sterile mice previously treated with antibiotics for 4 weeks. These mice were categorized into two groups: the Vehicle group (n=7) received fecal samples from individuals with normal bone mass, and the FMT group (n=7) received fecal samples from individuals with osteoporosis. After 8 weeks, bone mass, intestinal microbial composition, intestinal permeability and inflammation were assessed, followed by a correlation analysis. Results The bone mass was significantly reduced in the FMT group. Microbiota sequencing showed that Shannon index (p < 0.05) and Simpson index (p < 0.05) were significantly increased in Op groups, and β diversity showed significant differences. the recipient mice were similar. linear discriminant analysis effect size (LEfSe) analysis of mice showed that Halobiforma , Enterorhabdus , Alistipes , and Butyricimonas were significantly enriched in the FMT group. Lachnospiraceae and Oscillibacter were significantly enriched in the Vehicle group. H&E staining of intestinal tissues showed obvious intestinal mucosal injury in mice. Intestinal immunohistochemistry showed that the expression of Claudin and ZO-1 in the intestinal tissue of the FMT group mice was decreased. The FITC-Dextran (FD-4) absorption rate and serum soluble CD14 (sCD14) content were increased in FMT mice. Correlation analysis showed that these dominant genera were significantly associated with bone metabolism and intestinal permeability, and were associated with the enrichment of specific enzymes. Serum and bone tissue inflammatory cytokines detection showed that the expression of TNF-α and IL-17A in the FMT group were significantly increased. Conclusion Overall, our findings suggested gut microbiota from postmenopausal osteoporosis patients accelerate bone mass loss in mice. Aberrant gut microbiota might play a causal role in the process of bone mass loss mediated by inflammation after the destruction of the intestinal barrier.

The study of extracellular vesicles has become an incredibly important field of study, but the inherent heterogeneity of these vesicles continues to make their study challenging. The genetic variability and well-documented protocols for the growth and vesicle isolation from Leishmania parasites provide a unique opportunity to compare the heterogeneity of different populations secreted by Leishmania clones. Leishmania mexicana was cultured on solid SDM agar plates and 8 clonal colonies were selected. The EVs collected from the liquid cultures of these 8 clones were assessed by NTA, TEM, and proteomic analysis. We found that all 8 clonal L. mexicana cultures were visually indistinguishable from each other and had similar growth rate, and these physical similarities extended to their EVs. However, proteomic analysis reveals that the EVs collected have unique protein profiles compared to each other and EVs isolated from a heterogeneous liquid culture of L. mexicana . We selected 3 clonal EVs for further mouse infection experiments and found that EVs from CL7 L. mexicana consistently caused reduced footpad swelling in C57BL6 mice footpads compared to EVs from CL1, CL8, and heterogenous L. mexicana . This trend was not observed when infecting Balb/C mice and C57BL6 with the parasites alone, with only CL1 L. mexicana causing significantly increased infection in Balb/c mice. Our results together show that EVs isolated from different clonal colonies of L. mexicana have distinct differences in protein cargo which can lead to varying outcomes on Leishmania infection. Further evaluation will be needed to determine the underlying mechanisms behind this and verify that differences observed in infectivity are directly caused by variations between our L. mexicana clones, especially genetic sequencing and immunoblotting to validate our results.

Enteropathogenic Escherichia coli (EPEC) strains are subdivided into typical (tEPEC) and atypical (aEPEC) according to the presence or absence of a virulence-associated plasmid called pEAF. Our research group has previously demonstrated that two aEPEC strains, 0421-1 and 3991-1, induce an increase in mucus production in a rabbit ileal loop model in vivo . This phenomenon was not observed with a tEPEC prototype strain. Few studies on aEPEC strains evaluating their capacity to induce intestinal mucus hypersecretion were done. This study aimed to investigate aEPEC strains regarding their genotypic and phenotypic characteristics, their ability to alter mucus production in an in vivo intestinal infection model, and their potential mucinolytic activity. To investigate the relationship between strains 0421-1 and 3991-1 and 11 other aEPEC strains, their serotypes, sequence types (ST), and virulence factors (VF), several sequencing and genomic analyses were carried out. The study also involved researching the reproduction of mucus hypersecretion in rabbits in vivo . We found that the two mucus-inducing strains and two other strains (1582-4 and 2531-13) shared the same phylogroup (A), ST (378), serotype (O101/O162:H33), and intimin subtype (ι2), were phylogenetically related, and induced mucus hypersecretion in vivo . A wide diversity of VFs was found among the strains, confirming their genomic heterogeneity. However, among the genes studied, no unique virulence factor or gene set was identified exclusively in the mucus-inducing strains, suggesting the multifactorial nature of this phenomenon. The two strains (1582-4 and 2531-13) closely related to the two aEPEC strains that induced mucus production in vivo also induced the phenomenon. The investigation of the mucinolytic activity revealed that all aEPEC strains used mucins as their carbon sources. Ten of the 13 aEPEC strains could cross a mucin layer, and only four adhered better to agar containing mucin than to agar without mucin. The present study paves the way for subsequent investigations into the molecular mechanisms regarding cellular interactions and responses, as well as the correlation between virulence factors and the induction of mucus production/expression during aEPEC infections.

Background Advanced age is a primary risk factor for adverse COVID-19 outcomes, potentially attributed to immunosenescence and dysregulated inflammatory responses. In the post-pandemic era, with containment measures lifted, the elderly remain particularly susceptible, highlighting the need for intensified focus on immune health management. Methods A total of 281 elderly patients were enrolled in this study and categorized based on their clinical status at the time of admission into three groups: non-severe (n = 212), severe survivors (n = 49), and severe non-survivors (n = 20). Binary logistic regression analysis was employed to identify independent risk factors associated with disease severity and in-hospital outcomes. The diagnostic performance of risk factors was assessed using the receiver operating characteristic (ROC) curves. Kaplan-Meier survival analysis and log-rank test were utilized to compare the 30-day survival rates. Furthermore, the transcriptomic data of CD4 ⁺ T cells were extracted from Gene Expression Omnibus (GEO) database. Gene Set Enrichment Analysis (GSEA) was applied to reveal biological processes and pathways involved. Results In the comparison between severe and non-severe COVID-19 cases, significant elevations were observed in the neutrophil-to-lymphocyte ratio (NLR), C-reactive protein (CRP), and Serum Amyloid A (SAA) levels, concurrent with a notable reduction in CD8 ⁺ T cells, CD4 ⁺ T cells, natural killer (NK) cells, and monocytes (all p < 0.05). CD4 ⁺ T cells (OR: 0.997 [0.995-1.000], p <0.05) and NLR (OR: 1.03 [1.001-1.060], p <0.05) were independent risk factors affecting disease severity. The diagnostic accuracy for COVID-19 severity, as measured by the area under the curve (AUC) for CD4 ⁺ T cells and NLR, was 0.715 (95% CI: 0.645-0.784) and 0.741 (95% CI: 0.675-0.807), respectively. Moreover, patients with elevated NLR or IL-6 levels at admission exhibited significantly shorter survival times. Gene Set Enrichment Analysis (GSEA) revealed several biological pathways that are implicated in the regulation of immune responses and metabolic processes. Conclusions Lymphocytopenia and the cytokine storm onset are significant predictors of an unfavorable prognosis in elderly patients. The decrease in CD4 ⁺ T cells among elderly patients is detrimental to disease recovery, and the biological pathways regulated by these cells could potentially heighten vulnerability to SARS-CoV-2 infection, thereby exacerbating the development of associated complications.

Introduction The early prediction of sepsis based on machine learning or deep learning has achieved good results.Most of the methods use structured data stored in electronic medical records, but the pathological characteristics of sepsis involve complex interactions between multiple physiological systems and signaling pathways, resulting in mixed structured data. Some researchers will introduce unstructured data when also introduce confounders. These confounders mask the direct causality of sepsis, leading the model to learn misleading correlations. Finally, it affects the generalization ability, robustness, and interpretability of the model. Methods To address this challenge, we propose an early sepsis prediction approach based on causal inference which can remove confounding effects and capture causal relationships. First, we analyze the relationship between each type of observation, confounder, and label to create a causal structure diagram. To eliminate the effects of different confounders separately, the methods of back-door adjustment and instrumental variable are used. Specifically, we learn the confounder and an instrumental variable based on mutual information from various observed data and eliminate the influence of the confounder by optimizing mutual information. We use back-door adjustment to eliminate the influence of confounders in clinical notes and static indicators on the true causal effect. Results Our method, named CISepsis, was validated on the MIMIC-IV dataset. Compared to existing state-of-the-art early sepsis prediction models such as XGBoost, LSTM, and MGP-AttTCN, our method demonstrated a significant improvement in AUC. Specifically, our model achieved AUC values of 0.921, 0.920, 0.919, 0.923, 0.924, 0.926, and 0.926 at the 6, 5, 4, 3, 2, 1, and 0 time points, respectively. Furthermore, the effectiveness of our method was confirmed through ablation experiments. Discussion Our method, based on causal inference, effectively removes the influence of confounding factors, significantly improving the predictive accuracy of the model. Compared to traditional methods, this adjustment allows for a more accurate capture of the true causal effects of sepsis, thereby enhancing the model's generalizability, robustness, and interpretability. Future research will explore the impact of specific indicators or treatment interventions on sepsis using counterfactual adjustments in causal inference, as well as investigate the potential clinical application of our method.

Smart innovative nanocomposites based on active ingredients and metallic nanoparticles with effective wound healing and antifungal properties are efficient in overcoming the limitations of traditional therapeutic products. Open wounds provide an ideal niche for colonization by Candida albicans ( C. albicans ) which poses substantial global health issues owing to delayed wound healing and disordered healing mechanisms. Therefore, proficient innovative therapies that control C. albicans infection and promote wound healing are of imperative importance for the management of wounds and prevention of infection and possible complications. This study aims to design a novel nanocarrier platform based on a hydrogel loaded with silver nanoparticles (AgNPs) and doped with pomegranate peel extract (PPE) and hyaluronic acid (HA), offering an unprecedented opportunity to achieve skin repair and manage C. albicans colonization with an efficient wound healing process. Sprague-Dawley rats (n=100) were assigned to 5 groups and infected with C. albicans and distributed as follows: control positive (untreated) and four cutaneous wound-healing model groups treated topically with commercial cream and PPE-HA-AgNPs at full, 50%, and 25% concentrations for 15 days, respectively. Our findings revealed that the severity of clinical signs, C. albicans burden, and the expression of biofilm-related genes ALS1 , HYR1 , and PLB1 were diminished following treatment with PPE-HA-AgNPsIII. Notably, the formulated nanocomposite was very effective in extending the release of PPE-HA-AgNPs in infected wounds with retention percentages of 65.4% for PPE-HA-AgNPsIII. Topical administration of PPE-HA-AgNPsIII successfully alleviated the extensive inflammatory response and healed wounded skin via downregulation of tumor necrosis factor-alpha (TNF-α), interleukin-6 and IL-1 beta, and nitric oxide synthase (NOS) levels as shown by enzyme-linked immunosorbent (ELISA) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays. Interestingly, PPE-HA-AgNPsIII modulated angiogenic and wound healing markers as evidenced by the downregulation of MMP-9 and the upregulation of angiopoietin-1 (Ang-1), vascular endothelial growth factor (VEGF) (up to 10 days post-treatment), transforming growth factor-beta 1 (TGF-β1), bFGF , EGF, Ki- 67, and collagen I and III with efficient wound closure capability. This was evidenced by the lessening of histopathological severity, which accelerated the healing of the infected skin wounds post-treatment with PPE-HA-AgNPs. Overall, our formulated PPE-HA-AgNPs provide an effective innovative therapeutic strategy for the treatment of cutaneous wounds infected with C. albicans with maximized wound healing efficacy, indicating their potential in clinical practice.

We present this critical review with the aim of highlighting the current status of periodontal diseases, focusing on the relevance of host modulating agents and immune pathways, in addition to new complementary therapeutic approaches for the treatment of these pathologies. Periodontal diseases are prevalent pathologies worldwide and the main cause of edentulism in the adult population. Their pathogenesis seems to be based on a dysbiosis of the oral microbiota that interacts with the host’s immune defenses and is responsible for the inflammatory/immune response, which would be modified by a number of conditions such as individual susceptibility, environmental and sociodemographic factors, certain systemic pathologies and the individual’s genetic condition, among others. Numerous studies have reported on the complex web of inflammatory mediators in periodontal disease and their role in tissue destruction as well as in homeostatic imbalance. Precisely, the role of epigenetics as a modifier of the host genetic condition has captured research attention in recent years. Therefore, this mini-review first discusses an updated etiological hypothesis of periodontal disease and the roles of certain cytokines in the immune response. In addition, the latest therapeutic trends with new developments and future perspectives are summarized.

Introduction Viral infection usually stimulates a variety of host cell factors to modulate the life cycle of the virus. PIM1, a serine/threonine protein kinase widely involved in cell proliferation, survival, differentiation and apoptosis, was recently reported to be upregulated by Zika virus (ZIKV) infection. However, how ZIKV-PIM1 interactions affect the viral life cycle are not fully understood. Methods and results Here, we demonstrated that ZIKV replication was suppressed by the PIM1 kinase inhibitor SGI-1776 in both wt and Ifnar1 -/- murine peritoneal macrophages, indicating that PIM1 functions independently of type I IFN signaling. Co-immunoprecipitation and GST pull-down assays revealed that the ZIKV structural protein precursor membrane (prM) interacted with PIM1. Moreover, we found that prM protein stability was enhanced by PIM1, which was attributed to its kinase activity. Mechanistically, we revealed that prM can undergo ubiquitin‒mediated proteolysis and the E3 ubiquitin ligase AMFR can target prM for degradation. Importantly, PIM1 catalyzed phosphorylation of prM at Ser101 and Thr107, and this phosphorylation prevented the proteasome-dependent degradation of prM by impairing its association with AMFR. Therefore, the S101/T107-D phosphorylation mimic mutant of prM was more resistant to PIM1-induced increases in cellular abundance. Discussion These findings revealed PIM1 as a critical host factor that is advantageous to ZIKV and revealed that targeting the PIM1‒prM axis is a conducive strategy for controlling ZIKV infection.

Antimicrobial resistance is among the greatest threats to public health globally, and drug repurposing strategies may be advantageous to addressing this problem. Mefloquine, a drug traditionally used to treat malaria, has emerged as a promising antibiotic adjuvant, due to its ability to enhance the effectiveness of conventional antibiotics against resistant bacterial strains. In this paper, we first outline the enhancement properties of mefloquine and its mechanisms of action as an adjuvant antibiotic against multidrug-resistant bacteria. Mefloquine exhibits synergistic bacteriostatic effects when combined with colistin, β-lactams, antituberculosis drugs, quinolones, and linezolid. Potential mechanisms underlying its synergistic effects include inhibition of antibiotic efflux, disruption of bacterial cell membrane integrity, and disturbance of biofilm formation. In addition, we explore the bacteriostatic effects of several mefloquine derivatives against Mycobacterium tuberculosis and some fungi. Further, we summarize the findings of recent studies on other aspects of mefloquine activity, including its antiviral and antitumor effects. Finally, the advantages and challenges of mefloquine use as an antibiotic adjuvant in combination with antibiotics for bacterial inhibition are discussed. Overall, mefloquine shows excellent potential as an antibiotic adjuvant therapy against multidrug-resistant bacteria and is a promising candidate for combination therapy; however, further studies are needed to fully elucidate its mechanism of action and address the challenges associated with its clinical application.

Introduction Coronavirus disease 2019 (COVID-19) alters the gut microbiome. This study aimed to assess the association between the disease severity of COVID-19 and changes in stool microbes through a seven-month follow-up of stool collection. Methods We conducted a multicentre, prospective longitudinal study of 58 COVID-19 patients and 116 uninfected controls. Differences in the gut microbiota were analysed using 16S ribosomal RNA sequencing. The first stool samples were collected at an early convalescent phase of COVID-19, and the second sample was collected at least seven months after COVID-19 infection. Results and discussion At the order level, Eubacteriales and Bifidobacteriales decreased, while Bacteroidales and Burkholderiales increased in the COVID-19 group compared to the controls. Alpha diversity also decreased in COVID-19 patients compared to controls, with imperfect recovery of the gut microbiome after seven months. The compositional change in the gut microbiome between the early and late convalescent phases was largest in the moderate and severe groups. The severity of COVID-19 was the most influential clinical variable for microbiome composition (Sum of Sqs = 0.686, P = 0.006), and its effect persisted even after partialling out other effects such as antibiotic use and age. Thus, our study indicates a possible interaction between respiratory viral infection and the composition of the gut microbiota community, warranting future mechanistic and prospective longitudinal studies. Additionally, we were able to detect microbiome changes in patients who were re-infected with SARS-CoV-2. Notably, the dominant bacteria in the re-infected group were Lachnospiraceae and Faecalimonas umbilicata, compared to the one-time infected group.

Objective This study aimed to assess the efficacy of innovative Chemiluminescence Immunoassay (CLIA) in testing Cryptococcal Antigen (CrAg) across two medical centers, employing the FDA-approved CrAg Lateral Flow Assay (LFA) by IMMY as a reference standard. Methods The study encompassed patients diagnosed with cryptococcosis at West China Hospital of Sichuan University (HX) between July 2022 and May 2023, and Suzhou Fifth People’s Hospital (SZ) from September 2020 to September 2023. All specimens underwent simultaneous detection using the LFA (IMMY, Norman, USA) and CLIA (Chuanglan, Suzhou, China). Results A total of 628 patients were enrolled, revealing a remarkable 99.20% concordance between LFA and CLIA (623/628, 99.20%). The LFA exhibited a sensitivity of 96.83% (244/252) and specificity of 98.35% (179/182). Among the 42 patients with unaltered CrAg titers, the changes of Signal-to-Cut-Off ratio (ΔS/CO) results exhibited a noteworthy discrepancy, with 71.43% (30/42) demonstrating a decreasing trend in ΔS/CO of at least 10%. Conclusions The CLIA method demonstrated commendable specificity and sensitivity, exhibiting a high level of agreement with the FDA-approved LFA method. Additionally, CLIA demonstrated superior utility for treatment monitoring compared to LFA, offering continuous insight into the fluctuation of CrAg concentrations.

Influenza A virus poses significant public health challenges due to its high mutation rate and zoonotic potential. Whole-genome sequencing (WGS) is crucial for monitoring and characterizing these viruses. Oxford Nanopore Technologies (ONT) and Illumina next-generation sequencing platforms are commonly used, with ONT being advantageous for its long-read capabilities, portability, and unique ability to access raw data in real-time during sequencing, making it suitable for rapid outbreak responses. This study optimizes the ONT Ligation Sequencing Influenza A Whole Genome protocol by refining RT-PCR kits, primers, and purification methods, and evaluating automation for high-throughput processing. The alternative RT-PCR kits, combined with alternative primers, significantly improved read depth coverage and reduced short, untargeted reads compared to the original ONT protocol. The improvement was particularly evident in the minimum read depth coverage of polymerase segments, which often face challenges with achieving uniform coverage, displaying higher coverage at the 5’ and 3’ termini, and lower coverage in the central regions. This optimized protocol for targeted influenza A WGS not only enhances sequencing quality and efficiency, but is applicable to all NGS platforms, making it highly valuable for studying influenza adaptation and improving surveillance. Additionally, this protocol can be further refined and adapted for the sequencing of other pathogens, broadening its utility in various pathogen monitoring and response efforts.

Introduction Chronic endometritis (CE) is a prolonged, mild inflammation of the endometrial lining. This study investigated the impact of the impact of antibiotic treatment combined with vaginal Lactobacillus on pregnancy outcomes in infertile patients with CE during frozen embryo transfer (FET) cycles. Methods A retrospective analysis was performed on the clinical data of 7,385 patients who underwent FET. After applying the inclusion and exclusion criteria, 254 patients diagnosed with CE were eligible for inclusion. Of these, 119 patients received antibiotics alone, whereas 135 were treated with a combination of doxycycline and vaginal Lactobacillus. All patients underwent embryo transfer within 6 months following treatment. The general characteristics and pregnancy outcomes of the first FET cycle post-treatment were compared between the two groups. Results There were no statistically significant differences between the two groups in terms of general characteristics, clinical pregnancy rate, early miscarriage rate, and ectopic pregnancy rate. Patients who received a combination of doxycycline and Lactobacillus showed a higher biochemical pregnancy rate compared to those who received doxycycline alone, though this difference was not statistically significant (70.37% vs. 64.71%, P=0.313). Furthermore, the incidence rate of premature rupture of membranes was lower in the doxycycline-Lactobacillus group than in the doxycycline group (50.00% vs 33.33%, P=0.037). Conclusions Although this study observed the potential benefits of the antibiotic and vaginal probiotic treatment regimen in increasing the biochemical pregnancy rate and reducing the incidence of premature rupture of membranes, the current findings are insufficient to recommend the combined use of antibiotics and vaginal Lactobacillus as an intervention to improve reproductive outcomes in infertile patients with CE.

This retrospective multicenter study was conducted across 83 intensive care units (ICUs) in 16 cities in Sichuan, China. Critically ill patients diagnosed with heatstroke and lung infections were included in the study. Specimens from the lower respiratory tract were collected for microbiological testing, and the characteristics of the pathogens were described. A total of 462 patients diagnosed with heatstroke-related pulmonary infections were included, 134 patients (29.0%) tested positive for respiratory pathogens. The most frequently isolated strain was Klebsiella pneumoniae (34.3%), followed by Escherichia coli (28.4%), Staphylococcus aureus (20.9%). The results revealed that in the hyperthermic resistance group, there was a higher proportion of Pseudomonas aeruginosa [14(34.1%) vs 11(11.8%), p=0.002] and Stenotrophomonas maltophilia [4(9.8%) vs 1(1.1%), p=0.030] compared to the hyperthermic control group. And a higher proportion of Staphylococcus aureus [27(29.7%) vs 1(2.3%), p<0.001], were obtained during the earlier stages with elevated temperatures. Patients with Klebsiella pneumoniae (38.3 ± 1.9°C), Staphylococcus aureus (38.5 ± 2.2°C), and Pseudomonas aeruginosa (38.7 ± 1.9°C) exhibited a higher temperature environment. Our study provides crucial insights into the lower respiratory tract pathogenesis of heatstroke patients, identifying key pathogens and their temperature-dependent characteristics, thus providing a foundation for future empirical treatment strategies in heatstroke.

Background The COVID-19 pandemic and the resulting non-pharmaceutical interventions (NPIs) have led to changes in the epidemiology of other respiratory pathogens. This study was conducted to explore the epidemiological characteristics of 13 respiratory pathogens, including 11 respiratory viruses and 2 non-classical microorganisms, in hospitalised patients with acute respiratory tract infections (ARTIs) and to compare the prevalence of respiratory pathogens during and after the COVID-19 pandemic. Methods We conducted a single-centre retrospective study involving 8979 patients with ARTIs in Shijiazhuang City from December 2019 to December 2023. The GeXP analysis platform and multiple reverse transcription–PCR (mRT–PCR) technology were used to simultaneously detect 13 respiratory pathogens. The ARIMA model was constructed to predict the pathogen detection rate in each quarter of Shijiazhuang City in the next 2 y. Results Among the 8979 patients, 4169 (46.43%) tested positive for respiratory pathogens. The total pathogen detection rate rebounded in the year after the COVID-19 pandemic. After the COVID-19 pandemic, the positive rates in men were slightly higher than those in women and the positive rates in spring and winter were significantly higher than those in summer. The dominant pathogens during the COVID-19 pandemic were Influenza A viru (InfA; 24.08%) and Human Rhinovirus (HRV; 21.77%), and after the COVID-19 pandemic were InfA (27.92%) and H3 (21.17%). During the COVID-19 pandemic, InfA and HRV frequently occurred in all age groups. After the COVID-19 pandemic, InfA and Seasonal Influenza virus H3N2 (H3) frequently occurred in all age groups. Conclusions A series of NPIs introduced by the Chinese government during the COVID-19 pandemic had a significant impact on acute upper respiratory pathogenic infections. After the withdrawal of the NPIs, the spectrum of respiratory pathogens changed.

Background The rise of antibiotic resistance imposes the search for novel antimicrobial strategies as natural products or its combination with antibiotics. This study investigates the synergistic effects of terpenoids from Cissampelos oppositifolia (C. oppositifolia) essential oil in combination with antibiotics against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The aims were to evaluate the antimicrobial efficacy, analyze functional group modifications and assess molecular interaction. Methods Essential oil was extracted from C. oppositifolia by hydro-distillation. The EO was analyzed for terpenoid content via Thin Layer Chromatography (TLC). Antimicrobial activity was assessed using the disc diffusion method and Minimum Inhibitory Concentration determinations (MIC) by broth dilution followed by bactericidal essay (Time-killing). FTIR and UV spectroscopy were employed to detect functional group modifications in terpenoid-antibiotic combinations. Molecular docking studies assessed interaction energies between terpenoids and antibiotics. Results TLC identified α-pinene, δ-carene, and caryophyllene in the EO. δ-Carene exhibited the highest synergy with antibiotics, showing the lowest MIC of 0.04 mg/mL against S. aureus ATCC-43300 and 0.05 mg/mL against E. coli MTCC-739. Time-kill assays demonstrated that α-pinene, δ-carene, and caryophyllene achieved complete bacterial eradication by 4 hours in combination with amoxicillin against E. coli, and by 2 hours against S. aureus in combination with erythromycin. FTIR analysis revealed peak shifts at 1599, 1774, and 2259 cm⁻¹ for amoxicillin + α-pinene, and new peaks at 1648 and 1287 cm⁻¹ for δ-carene + erythromycin. UV spectra indicated potential complex formations. Docking studies showed δ-carene’s strong interaction with erythromycin and amoxicillin, with interaction energies of -96.10 and -87.75 kcal/mol, respectively. Conclusion Terpenoids from C. oppositifolia enhance the antimicrobial efficacy of antibiotics. Functional group modifications and complex formations suggest that these interactions may contribute to synergistic effects. These findings support the potential use of terpenoid-antibiotic combinations in overcoming antibiotic resistance and warrant further investigation into their mechanisms of action.

Background Systemic inflammatory indicators such as neutrophil-to-lymphocyte ratio (NLR) can effectively predict the prognosis of various inflammatory diseases. However, its prognostic effect on patients with carbapenem-resistant Klebsiella pneumoniae (CRKP) infection is little known. The objective of this study was to investigate the risk factors for mortality associated with CRKP infection and the clinical value of NLR in predicting prognosis in these patients. Methods A total of 190 inpatients with CRKP infection from 1 January 2023 to 31 December 2023 were enrolled in this study, namely, 73 fatal cases and 117 survival cases in hospital. The medical data and examination results of these patients were collected. A logistic regression analysis was performed to assess the association between the NLR on the day of CRKP infection onset and all-cause mortality in hospital. Results The overall mortality rate of patients with CRKP infection was 38.42% (73/190). Of the 190 patients, 91 were co-infected with carbapenem-resistant Acinetobacter baumannii/carbapenem-resistant Pseudomonas aeruginosa (CRAB/CRPA). Multifactor regression analysis confirmed that carbapenem exposure in the past 14 days, central line insertion, and chronic Foley catheter requirement were independent risk factors for carbapenem-resistant bacteria co-infection. The multivariate analysis shows that admission to an ICU, co-infection with CRAB/CRPA, and higher NLR were independent risk factors for the mortality in hospital, while appropriate treatment within 3 days was an independent protective factor. The area under the curve (AUC) of the NLR was 0.696, and the cutoff value of the NLR was 10.73. Conclusions The NLR on the day of CRKP infection onset, admission to an ICU, and co-infection with CRAB/CRPA were identified as independent risk factors for all-cause mortality of patients with CRKP infection, while appropriate treatment within 3 days was recognized as an independent protective factor. The NLR serves as a conveniently accessible and independent prognostic biomarker for patients with CRKP infection.

Background This study aims to develop a risk prediction model for multidrug-resistant bacterial and fungal infections in patients with gastrointestinal fistulas during the perioperative period. Methods A retrospective cohort study was conducted at Anhui No. 2 Provincial People’s Hospital from January 2022 to July 2024. We analyzed the distribution, resistance patterns, and mechanisms of multidrug resistance. Univariate and multivariate logistic regression analyses were performed to identify independent risk factors. A nomogram was constructed based on these risk factors, and its performance was evaluated using calibration curves, receiver operating characteristic (ROC) curves, and decision curve analysis (DCA). Results A total of 266 patients were included, with 157 (59.02%) testing positive for multidrug-resistant infections. We isolated 329 pathogenic strains: 84 Gram-positive (25.53%), 215 Gram-negative (65.35%), and 30 fungal strains (9.11%). The most common isolate was Klebsiella pneumoniae (57 strains, 17.33%). Patients were divided into a training cohort (n = 177) and a validation cohort (n = 89). Multivariate analysis identified six key indicators: secondary surgery, length of hospital stay, preoperative white blood cell (WBC) count, preoperative neutrophil count, postoperative WBC count, and postoperative C-reactive protein (CRP) levels. The nomogram demonstrated excellent predictive ability, with an area under the curve (AUC) of 0.905 in the training cohort and 0.793 in the validation cohort. Calibration curves indicated high consistency between predicted probabilities and observed values. DCA confirmed the clinical utility of the nomogram. Conclusion Our study shows that multidrug-resistant infections in patients with gastrointestinal fistulas are predominantly caused by Gram-negative bacilli, especially carbapenem-resistant Enterobacteriaceae. Key risk factors include secondary surgery and various blood count parameters. The developed nomogram provides robust predictive accuracy, aiding healthcare providers in implementing targeted infection prevention strategies.

Introduction Traditional Chinese Medicine (TCM), specifically Gui-Shen-Wan, has shown promise in restoring ovarian function among reproductive-age women who had impaired ovarian functions, yet the underlying mechanisms remain elusive. Recent studies highlight the pivotal role of the gut microbiome (GM) in mediating the therapeutic effects of TCM. However, it is unclear whether the GM contributes to Gui-Shen-Wan’s therapeutic restoration of ovarian functions. Methods This study employed a mouse model with cyclophosphamide-induced decreased ovarian function (P_T and P_AT groups) and a control group without modeling. The P_AT group received a 7-day course of oral antibiotics to deplete the GM prior to a 20-day Gui-Shen-Wan treatment regimen. Results Both P_T and P_AT mice exhibited prolonged metestrus/diestrus phases compared to controls (p<0.05), indicating menstrual disruption post-modeling. Following 20 days of Gui-Shen-Wan treatment, P_T mice showed a shorter metestrus/diestrus phase (4 days) compared to P_AT mice (5 days) (p<0.05). Notably, P_T mice had a higher number of normal follicles(primitive/primary/secondary/antral follicles) in their ovaries post-treatment (median 15) compared to P_AT mice (median 8.5). Proteome analysis revealed that ovarian proteins enriched in P_T mice were primarily associated with oxidative phosphorylation and DNA replication pathways, suggesting GM-mediated enhancement of these processes. Discussion This study underscores the pivotal role of the GM in the therapeutic benefits of Gui-Shen-Wan, highlighting the potential for microbiome-targeted interventions in promoting beneficial effects of Gui-Shen-Wan on the restoration of decreased ovarian functions.