Although China has successfully eliminated neonatal tetanus, non-neonatal tetanus, particularly among older adults, remains a persistent clinical and public health challenge. This narrative review synthesizes the current landscape of tetanus in China—covering epidemiology, pathophysiology, prevention, diagnosis, and treatment—and compares it with World Health Organization (WHO) guidance and practices in other countries to identify gaps and optimization strategies. A comprehensive narrative review was conducted across five core domains: epidemiology, pathogen and pathophysiology, prevention strategies, diagnostic advances, and therapeutic interventions. Data were integrated from the WHO, the Chinese Center for Disease Control and Prevention, national guidelines, and relevant clinical studies, with comparative analysis against global best practices. Epidemiological trends show a clear shift in disease burden from neonates to older adults, predominantly in rural areas, with high case fatality rates (10–40

This study presents a case of Ralstonia mannitolilytica isolated from a blood culture of a 50-year-old male patient with hepatocellular carcinoma. The patient underwent ultrasound-guided radiofrequency ablation for liver lesions associated with hepatocellular carcinoma and subsequently experienced recurrent fever, chills, and rigors. R. mannitolilytica was identified from blood culture. This bacterium, known to be an opportunistic pathogen, has not been extensively studied for its whole-genome characteristics and resistance profiles. Through whole-genome sequencing and bioinformatics analysis, we identified 32 distinct drug resistance genes associated with various antibiotic classes, including beta-lactams, aminoglycosides, and fluoroquinolones. Our findings reveal the potential for multidrug resistance in R. mannitolilytica, which exhibited numerous resistance genes. This underscores the clinical challenge of treating infections caused by this bacterium and highlights the importance of whole-genome analysis in understanding emerging pathogens. The identification of these resistance genes is crucial for devising effective clinical strategies to combat infections, particularly in immunocompromised patients, in whom the bacterium may pose a significant risk.

Purpose:This study aims to investigate the antibiotic resistance and virulence factors of Paenibacillus campinasensis, an understudied opportunistic pathogen, through whole-genome sequencing and resistance genes. The findings will contribute to the development of effective clinical strategies for infection control. Patients and Methods:A 1-year and 3-month-old boy presenting with fever and rash was admitted to the hospital and clinically diagnosed with hand-foot-mouth disease. A blood culture was obtained, and the pathogen was preliminarily identified as belonging to the genus Paenibacillus using MALDI-TOF-MS. Subsequent 16S rDNA PCR product Sanger sequencing confirmed the isolate as Paenibacillus campinasensis. Whole-genome sequencing and bioinformatics analysis were performed to evaluate the antibiotic resistance and virulence factors of the strain. Results:Whole-genome sequencing and bioinformatics analysis identified 18 resistance genes in the Paenibacillus campinasensis strain, spanning multiple antibiotic categories. These findings suggest the potential for multidrug resistance in this bacterium, despite its isolation from blood without associated clinical infection in the patient. Conclusion:The identification of resistance genes in Paenibacillus campinasensis underscores the challenges in clinical treatment and emphasizes the critical role of whole-genome analysis in pathogen detection and characterization. These findings are vital for informing effective clinical strategies to address infections, particularly given the potential for antibiotic resistance to hinder treatment outcomes, even in the absence of direct disease causation by the bacterium.

Acinetobacter baumannii is a notable opportunistic pathogen responsible for severe hospital-acquired infections, with multidrug-resistant strains posing significant treatment challenges. Phage therapy, which employs bacteriophages as natural bacterial antagonists, has gained renewed attention as a promising solution to combat antibiotic-resistant infections. In this study, we isolated and characterized a novel virulent phage, vB_AbaS_qsb1, which specifically lyses A.baumannii. Phylogenetic and genomic analyses indicate that vB_AbaS_qsb1 is the founding member of a previously unreported genus, which we propose to name Acinibactriovirus, with Acinibactriovirus lysinus as the type species. vB_AbaS_qsb1 demonstrated robust stability across diverse temperature and pH ranges, a short latent period, and no known virulence or antibiotic resistance genes within its 54,713 bp dsDNA genome. Safety assessments showed that high-dose vB_AbaS_qsb1 induced no adverse effects in mice, with histopathology confirming its safety profile. Therapeutic experiments further indicated that vB_AbaS_qsb1 provided at least 50% protection against A.baumannii-induced pneumonia, significantly reducing bacterial loads and inflammation markers, while maintaining high phage titers in lung tissue.This study introduces vB_AbaS_qsb1 as a promising candidate for phage therapy against A.baumannii, offering both innovative insights and a valuable framework for future isolation, genomic characterization, and efficacy evaluation of phages targeting antibiotic-resistant bacteria.

Acute respiratory infections, caused by RNA viruses like respiratory syncytial virus, influenza, rhinovirus, and coronavirus, are major global health threats. Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) is the gold standard for detecting these viruses but is time-consuming, complex, and requires specialized equipment. There is a need for rapid, convenient, and multi-target detection methods to improve disease prevention and control. This study developed a multi-target immunochromatographic detection method using LbuCas13a protein and "band elimination" test strips for detecting SARS-CoV-2 and influenza virus. The method's performance was evaluated by testing known 5 positive and 4 negative samples for SARS-CoV-2 and comparing results with fluorescent PCR and colloidal gold methods. Detection sensitivity was quantified using digital PCR and qPCR. The immunochromatographic test strips showed 100% concordance with fluorescent PCR and colloidal gold methods in initial clinical SARS-CoV-2 detection. Subsequently, we used dual-target immunochromatographic test strips to detect 9 SARS-CoV-2 positive samples and 9 H3N2 positive samples. However, false negatives were observed in dual-target detection of SARS-CoV-2 and H3N2 samples, likely due to low sample concentration or sample degradation. The method had a minimum detection limit of 381.75 copies/µL, as determined by digital PCR and qPCR. The developed multi-target immunochromatographic detection method offers a rapid, low-cost, and simple approach for detecting both SARS-CoV-2 and influenza viruses. With high sensitivity, specificity, and reliability, this method holds promise as a practical tool for RNA virus diagnosis and improving public health response to respiratory infections.

Carotid and femoral plaques exhibit varying degrees of stability; however, the relationships of different genes/cell types with plaque embolism are poorly understood. We evaluated differential gene/cell expression and investigated the cells/genes associated with carotid and femoral artery plaque embolism. sc-RNA-seq and bulk RNA data were obtained to identify differentially expressed genes (DEGs). Seven machine learning models were trained, and the top 10 DEGs across all models were selected. The most disturbed cells in carotid and femoral artery plaques were identified using Augur, while the genes and cells in the carotid plaque associated with embolism were analyzed through scPagwas. The differences in most disturbed cells and embolism-related cells were further analyzed. Compared with femoral plaques, carotid plaques had 80 downregulated and 90 upregulated genes. Machine learning identified the key DEGs between carotid and femoral plaques were predominantly from the HOX gene family. Natural Killer (NK) cells were the most significantly disturbed cells between carotid and femoral plaques, and they may be most strongly associated with plaque embolism. Among the differential genes in NK cells, CD2 was most associated with embolism. Our research may offer new insights into atherosclerosis at different locations.

BACKGROUND:Pulmonary infections are prevalent opportunistic complications among HIV/AIDS patients, necessitating precise pathogen identification for targeted treatment. The traditional culture methods used for diagnosing these infections have inherent limitations when it comes to detecting the diverse microbial flora present in HIV/AIDS patients. METHODS:In this study, sputum samples were collected from 37 HIV/AIDS patients admitted to Mengchao Hepatobiliary Hospital of Fujian Medical University. A total of 45 samples were subjected to analysis using three different methods: traditional culture methods, third-generation long-read sequencing (PacBio), and droplet digital PCR (ddPCR). The study aimed to compare the diagnostic capabilities and accuracy of microorganism identification among these methods. RESULTS:Out of 45 sputum samples, the traditional culture method identified only 5 positive cases, whereas PacBio sequencing detected all samples positively. Among the 7 samples (including 5 traditional culture positive and 2 negative samples), PacBio sequencing was able to identify 51 species (77 strains), while ddPCR detected 15 species (64 strains). In contrast, traditional culture methods were only able to identify 3 bacterial species (6 strains). In samples that tested positive by culture, both PacBio sequencing and ddPCR were consistent in identifying the primary microorganisms, indicating the presence of multiple concurrent infections. In samples that were negative by culture, both PacBio and ddPCR were able to detect a broader range of microorganisms, showing a higher sensitivity compared to traditional methods. PacBio sequencing provided a comprehensive view of the microbial flora, whereas ddPCR enabled rapid and precise detection of common microorganisms and antibiotic resistance genes, compensating for PacBio's shortcomings in absolute quantification. CONCLUSIONS:The study demonstrates that PacBio sequencing and ddPCR offer significant advantages over traditional culture methods in diagnosing pulmonary infections in HIV/AIDS patients. These advanced techniques not only detect a greater variety of pathogens but also provide a more detailed understanding of the microbial flora and resistance profiles, which is crucial for effective clinical management. The combination of PacBio's comprehensive profiling and ddPCR's rapid detection capabilities presents a powerful approach to overcoming the limitations of traditional diagnostic methods in this patient population.

Background Nasopharyngeal carcinoma (NPC) treatment is largely based on a ‘one-drug-fits-all’ strategy in patients with similar pathological characteristics. However, given its biological heterogeneity, patients at the same clinical stage or similar therapies exhibit significant clinical differences. Thus, novel molecular subgroups based on these characteristics may better therapeutic outcomes. Methods Herein, 192 treatment-naïve NPC samples with corresponding clinicopathological information were obtained from Fujian Cancer Hospital between January 2015 and January 2018. The gene expression profiles of the samples were obtained by RNA sequencing. Molecular subtypes were identified by consensus clustering. External NPC cohorts were used as the validation sets. Results Patients with NPC were classified into immune, metabolic, and proliferative molecular subtypes with distinct clinical features. Additionally, this classification was repeatable and predictable as validated by the external NPC cohorts. Metabolomics has shown that arachidonic acid metabolites were associated with NPC malignancy. We also identified several key genes in each subtype using a weighted correlation network analysis. Furthermore, a prognostic risk model based on these key genes was developed and was significantly associated with disease-free survival (hazard ratio, 1.11; 95% CI, 1.07–1.16; P < 0.0001), which was further validated by an external NPC cohort (hazard ratio, 7.71; 95% CI, 1.39–42.73; P < 0.0001). Moreover, the 1-, 3-, and 5-year areas under the curve were 0.84 (95% CI, 0.74–0.94), 0.81 (95% CI, 0.73–0.89), and 0.82 (95% CI, 0.73–0.90), respectively, demonstrating a high predictive value. Conclusions Overall, we defined a novel classification of nasopharyngeal carcinoma (immune, metabolism, and proliferation subtypes). Among these subtypes, metabolism and proliferation subtypes were associated with advanced stage and poor prognosis of NPC patients, whereas the immune subtype was linked to early stage and favorable prognosis.

Trauma is a significant health issue that not only leads to immediate death in many cases but also causes severe complications, such as sepsis, thrombosis, haemorrhage, acute respiratory distress syndrome and traumatic brain injury, among trauma patients. Target protein identification technology is a vital technique in the field of biomedical research, enabling the study of biomolecular interactions, drug discovery and disease treatment. It plays a crucial role in identifying key protein targets associated with specific diseases or biological processes, facilitating further research, drug design and the development of treatment strategies. The application of target protein technology in biomarker detection enables the timely identification of newly emerging infections and complications in trauma patients, facilitating expeditious medical interventions and leading to reduced post-trauma mortality rates and improved patient prognoses. This review provides an overview of the current applications of target protein identification technology in trauma-related complications and provides a brief overview of the current target protein identification technology, with the aim of reducing post-trauma mortality, improving diagnostic efficiency and prognostic outcomes for patients.

Glioma stem cells (GSCs) are thought to be responsible for the initiation and progression of glioblastoma (GBM). GBM presents highly invasive growth with a very high recurrence rate, so it has become a clinical problem to be solved urgently. RNAseq demonstrates that thrombospondin 1 (THBS1) acts not only in the angiogenic core of glioma but also with a high degree of invasiveness and infiltration. Nevertheless, defects in the signaling pathway research lead to a poor prognosis in glioma patients. To investigate the relevant molecular mechanism and signal pathway of glioma stem cell behavior mediated by THBS1, U251 astroglioma cells and GSCs were taken as model cells for in vitro experiments. The biological effects of THBS1 on glioma proliferation, migration, and adhesion were evaluated using Cell Counting Kit-8(CCK8) assays, EdU incorporation assays, migration assays, Transwell assays, Western blotting, and RNAseq. We found that the knockout of the THBS1 gene by CRISPR/Cas9 promoted proliferation and migration in U251 cells and GSCs, as well as influencing cell cycle progression by regulating the TNF/MAPK/NF-κB and TGF-β/Smad signaling pathways. Moreover, U251 cells and GSCs showed different responses to THBS1 knockout, suggesting specific and potential targets for GSCs in signaling pathways mediated by THBS1.

Breast cancer (BC) is a heterogeneous disease characterized by significant differences in prognosis and therapy response. Numerous prognostic tools have been developed for breast cancer. Usually these tools are based on bulk RNA-sequencing (RNA-Seq) and ignore tumor heterogeneity. Consequently, the goal of this study was to construct a single-cell level tool for predicting the prognosis of BC patients. In this study, we constructed a stemness-risk gene score (SGS) model based on single-sample gene set enrichment analysis (ssGSEA). Patients were divided into two groups based on the median SGS. Patients with a high SGS scores had a significantly worse prognosis than those with a low SGS, and these groups exhibited differences in several tumor characteristics, such as immune infiltration, gene mutations, and copy number variants. Our results indicate that the SGS is a reliable tool for predicting prognosis and response to immunotherapy in BC patients.

Both the Nuclear factor-erythroid 2 p45-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) antioxidant pathway and Nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) pathway are considered essential for the development of acute lung injury (ALI)/ARDS induced by sepsis. Our aim was to study the role of Nrf2/HO-1 pathway on activation of the NLRP3 in the protective effect of marrow mesenchymal stem cells (BMSCs) on LPS–induced ALI. We found that BMSCs ameliorated ALI as evidenced by 1) decreased histopathological injury, wet/dry ratio, and protein permeability index in lung; 2) decreased reactive oxygen species (ROS), malondialdehyde (MDA), and protein carbonyl content and restored the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in lung tissue; 3) reduced LPS–induced increase in inflammatory cell count and promotion of tumor necrosis factor (TNF)-α, interleukin (IL)–1β, and IL-6 levels in bronchoalveolar lavage fluid (BALF); 4) improvement in the four-day survival rate of animals; and 5) enhanced expression of Nrf2 and HO-1 and decreased expression of NOD-like receptor protein 3(NLRP3) and caspase-1 (p20) in lung tissue. Of note, Nrf2 transcription factor inhibitor brusatol and HO-1 inhibitor tin protoporphyrin IX (SnppIX) reversed BMSCs induced down-expression of NLRP3 and caspase-1 (p20), and inhibited the protective effects of BMSCs. These findings demonstrated that the Nrf2-mediated HO-1 signaling pathway plays a critical role in the protective effects of BMSCs on LPS-induced ALI. BMSCs may play an anti-inflammatory effect partly through the Nrf2/HO-1-dependent NLRP3 pathway.

EvolvR系统是一种CRISPR介导的新型定向进化技术,本研究探索其在酶基因序列定向进化中的适用性以及突变效率,同时在此基础上扩大EvolvR的突变窗口长度.研究成功将4个能高效表达的单个sgRNA串联,并检测到靶向同一基因的3个不同靶位点.证明EvolvR具有在目标基因区域大范围制造突变的潜力,具有很高的应用价值.为利用EvolvR系统对目标酶基因实现定向进化的研究奠定基础.

There is a lack of evidence about the relationship between microorganisms and non-carious cervical lesions (NCCLs) due to limited technologies. A group of 78 patients was enrolled for microbial 16S rRNA sequencing of dental plaques on normal and defective cervical surfaces. Parallel data from 39 patients were analysed with paired t tests, and Fusobacteriales exhibited significantly less distribution on NCCLs than on normal surfaces. As a result, Fusobacterium nucleatum, the most common oral bacterial strain belonging to the order Fusobacteriales, was selected for further research. From a scanning electron microscopy (SEM) scan, the tooth surface with Fusobacterium nucleatum and Streptococcus mutans culture was more intact than that without Fusobacterium nucleatum. Furthermore, the calcium contents in groups with Fusobacterium nucleatum were significantly higher than that without it. In further mechanistic research, Fusobacterium nucleatum was demonstrated to adhere to and disturb other organisms as well as producing alkaline secretions to neutralize the deleterious acidic environment, protecting the tooth structure. In conclusion, microorganisms and NCCLs were confirmed directly related through adherent bacterial interactions and pH regulation. The research provides a new perspective and experimental evidence for the relation between microorganisms and NCCLs, which guides clinical treatment and preventive dentistry in the future.

Banknotes have long been suspected to be biologically “dirty” due to their frequent human contact, which may transmit human microbial pathogens. Still, it is an unsettled issue whether the microbes on banknotes pose a real threat to human health. In several previous studies, metagenomic sequencing was used to reveal the diversities of microbes on banknotes but live microorganism culture and functional verification were lacking. In this study, we collected banknotes of RMB in China as well as dollar bills in the United States and analyzed the microbial biodiversity and drug resistance genes carried by the identified microbes by metagenomic sequencing and in vitro culture methods. We identified eight major genera of drug-resistant bacteria through screening of 30 antibiotics, and the blood agar plate culture uncovered six pathogenic fungal species. Numerous phage and six dangerous viral sequences were also found. These results should substantiate our concern about the potential risk of banknotes to human health.

Directed evolution is a cyclic process that alternates between constructing different genes and screening functional gene variants. It has been widely used in optimization and analysis of DNA sequence, gene function and protein structure. It includes random gene libraries construction, gene expression in suitable hosts and mutant libraries screening. The key to construct gene library is the storage capacity and mutation diversity, to screen is high sensitivity and high throughput. This review discusses the latest advances in directed evolution. These new technologies greatly accelerate and simplify the traditional directional evolution process and promote the development of directed evolution.

Traditional pathogenic diagnosis presents defects such as a low positivity rate, inability to identify uncultured microorganisms, and time-consuming nature. Clinical metagenomics next-generation sequencing can be used to detect any pathogen, compensating for the shortcomings of traditional pathogenic diagnosis. We report third-generation long-read sequencing results and second-generation short-read sequencing results for ascitic fluid from a patient with liver ascites and compared the two types of sequencing results with the results of traditional clinical microbial culture. The distribution of pathogenic microbial species revealed by the two types of sequencing results was quite different, and the third-generation sequencing results were consistent with the results of traditional microbial culture, which can effectively guide subsequent treatment. Short reads, the lack of amplification, and enrichment to amplify signals from trace pathogens, and host background noise may be the reasons for the high error in the second-generation short-read sequencing results. Therefore, we propose that long-read-based rRNA analysis technology is superior to the short-read shotgun-based metagenomics method in the identification of pathogenic bacteria.

As an important global medical problem, hepatocellular carcinoma (HCC) has been recognized as the most frequent primary liver cancer and a leading cause of death among patients with cirrhosis. Surveillance of HCC using serum markers aims to reduce the disease-related mortality of HCC. MUC16 (mucin 16, also known as carbohydrate antigen 125, CA125) has been predicted as a tumor biomarker for many cancer types. Based on the high frequency mutation rate in a database from the Cancer Genome Atlas (TCGA), we investigated the effects of MUC16 knockdown and the regulatory profile of MUC16 in HepG2 and Huh7 cell lines. Knockdown of MUC16 was conducted via siRNA transfection, and the proliferation of cells was not affected by CCK8 assay results. Moreover, decreasing the expression of MUC16 enhanced the migration and invasion of cells, as shown by wound healing and transwell assays. Furthermore, RNA-seq was used to investigate the effect of MUC16 knockdown on the gene expression profile of HepG2 and Huh7 cells. Our study demonstrated the significant role of MUC16 in the inhibition of the migration and invasion of HepG2 and Huh7 cells.

Diatom test is the most commonly used method to diagnose drowning in forensic laboratories. However, microscopic examination and identification of diatom frustules is time-consuming and requires taxonomic expertise. At present, the identification of drowning is still a challenge in forensic casework. In this study, we developed a novel diatom microarray based on the detection of specific 18S rRNA gene fragments of diatom species. The array covers 169 diatom species which were documented as commonly found in a wide range of fresh waters in China. Diatom arrays were prepared from species specific oligonucleotide probes targeting to variable regions of the 18S rRNA gene. We also developed an auxiliary sample preparation method for isolation of diatom DNA from tissues, which enabled detection of diatom species in real forensic samples as well as environmental waters. We applied the diatom arrays to analyze six drowned cases and eight environmental samples. The diatom arrays showed much better sensitivity and more consistent results than those of the conventional SEM methods. We discovered major discrepancies between results generated by the diatom arrays and the routinely used SEM based diatom tests. We verified the results of our diatom arrays by species specific PCR and Sanger sequencing and found that the currently used SEM diatom test method has a serious deficiency in sensitivity due to high loss rate of frustules in the sample preparation procedure. We anticipate that the application of diatom arrays will transform current forensic practice of diagnosing drowning deaths.

Circular RNAs (circRNAs), a new category of noncoding RNA, have emerged in recent years as novel biomolecules with important biological functions. Increasing evidence and reports have revealed that circRNAs play an important role in human carcinogenesis and tumor progression. Gastric cancer (GC) is one of the most prevalent life-threatening malignancies worldwide, and in the present study, a novel circRNA molecule (circRIMS) was shown to be associated GC metastasis using next-generation sequencing. CircRIMS remarkably promoted GC cell metastasis in vitro, functioning as a sponge for hsa-miR-148a-5p and hsa-miR-218-5p. In addition, the results of rescue experiments showed that hsa-miR-148a-5p and hsa-miR-218-5p mimics could reverse the tumor-promoting roles of circRIMS in GC. Thus, circRIMS has potential as an early biomarker for use in predicting invasive metastasis in GC and to guide clinical diagnosis and treatment for precision medicine.