This study aims to investigate the disinfection and sterilization effects of common disinfectants on common pathogens on smooth and porous surfaces in the environment, providing a basis for blocking the transmission routes of pathogens in the environment and formulating practical and effective disinfection technology pathways. In this experiment, the suspension quantitative sterilization method was used to detect the sterilization effects of 75% ethanol, 1% hydrogen peroxide, and 500 mg/L chlorine-containing disinfectant on common laboratory environmental bacteria (Escherichia coli and Staphylococcus aureus) in different scenarios such as smooth and porous surfaces. In smooth scenarios, 1% hydrogen peroxide disinfectant showed significantly better disinfection effects than 75% ethanol and chlorine-containing disinfectant. When the action time was 1～5 minutes, its logarithmic reduction value was greater than 5, meeting the disinfection qualification requirements. On porous surfaces, when the action time of chlorine-containing disinfectant was 1～5 minutes, it showed high logarithmic reduction values (≥4) against different pathogens. Field experiments also found that the disinfection effect of chlorine-containing disinfectant was better than that of 1% hydrogen peroxide and 75% ethanol. Chlorine-containing disinfectant has a good disinfection effect and acts quickly, making it suitable for rapid disinfection of different surface structures in various scenarios. However, it is not environmentally friendly. When choosing a rapid disinfectant, a comprehensive assessment must be made based on its harmfulness to the environment, among other factors.

This study aimed to analyze the distribution and drug sensitivity of pathogenic bacteria of bloodstream infections in patients hospitalized in Xinchang County Hospital of Traditional Chinese Medicine, providing a basis for the diagnosis and treatment of clinical bloodstream infections. All blood culture samples from Xinchang County Hospital of Traditional Chinese Medicine were collected from January 2019 to June 2023, and the pathogen species and drug sensitivity in vitro were analyzed statistically. A total of 1692 positive blood culture samples were obtained, primarily from the intensive care unit (ICU). The pathogens detected included 1637 aerobic (96.74%), 19 anaerobic (1.12%) and 36 fungal (2.13%) strains. The gram-negative bacteria (36.63%) were mainly Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis, and the gram-positive bacteria (63.12%) were mainly Staphylococcus hominis, Staphylococcus epidermidis, and Staphylococcus aureus. Drug sensitivity results indicated that Escherichia coli and Klebsiella pneumoniae exhibited resistance rates exceeding 70% to aztreonam but sensitivity rates exceeding 70% to cephalosporins. In contrast, Staphylococcus epidermidis, Staphylococcus aureus and Staphylococcus hominis showed resistance rates exceeding 70% to penicillin, but high sensitivity (greater than 90%) to antimicrobial agents such as linezolid and tigecycline. The positive blood culture samples are predominantly from wards housing immunocompromised patients. The primary pathogenic bacteria identified were Escherichia coli, Klebsiella pneumoniae, Staphylococcus hominis and Staphylococcus epidermidis, which are sensitive to some antibacterial drugs. However, the widespread use of antibiotics in treatment may lead to changes in drug resistance patterns. Therefore, clinical practices should emphasize the detection of pathogenic bacteria and their drug susceptibility profiles to guide the diagnosis and treatment of bloodstream infections.

This paper reports a case of pulmonary infection caused by Corynebacterium striatum in an elderly patient. The patient, male, 93 years old, was treated with piperacillin-tazobactam, imipenem-cilastatin and levofloxacin due to “aspiration pneumonia” two months ago. The patient’s pulmonary infection was difficult to control, and meropenem was upgraded for anti-infection, but the effect was not good. After empiric antifungal treatment with fluconazole, the pulmonary infection continued to persist and the symptoms worsened. Sputum culture result suggested Corynebacterium striatum (＋＋＋＋), and linezolid showed a large diameter inhibitory zone against the bacteria. Considering the previous treatment plan was not satisfactory, fluconazole was discontinued and the regimen was adjusted to meropenem combined with linezolid for anti-infection. Ten days after the adjustment of the antimicrobial agents, the patient’s symptoms of cough and sputum production gradually improved. Subsequently, meropenem was de-escalated to levofloxacin combined with linezolid for anti-infection. After 2 days of de-escalation therapy, the patient’s pulmonary infection improved, and the antimicrobial agents were discontinued, resulting in successful treatment of the pulmonary infection.

In order to understand the T cell immune status of active tuberculosis (ATB) patients, the study compared the expression levels of T cell immune molecules and the proportion of T cell functional subsets between ATB patients and healthy controls (HC). A total of 21 ATB patients and 10 HC were enrolled from December 2020 to May 2021. Peripheral blood samples were collected from the participants, and flow cytometry was performed for surface and intracellular staining to investigate the proportion and functional status of peripheral blood T cells. Fisher’s exact probability test and non-parametric Mann-Whitney U test were used to analyze the data. The result showed that the proportion of Tfh in CD4^＋ T cells was significantly lower in ATB patients than that in HC group (P<0.05), and the proportions of naive T cells, progenitor exhausted T cells in CD8^＋ T cells were lower in ATB patients compared to HC, while terminal exhausted T cells were higher (P<0.05). The expression of CD62L on CD4^＋ T cells was significantly lower in the ATB group (P＝0.01). The expression of TIM-3 and CD127 on CD8^＋ T cells as well as T-bet and TCF1 in CD8^＋ T cells in the ATB group were also significantly lower than those in the HC group, while KLRG1, PD1, TIGIT, and CD69 were higher (P<0.05). In conclusion, T cells from active tuberculosis patients exhibit a unique immune phenotype characterized by reduced pro-inflammatory capacity and increased anti-inflammatory capacity, meanwhile exacerbated differentiation towards an exhausted phenotype in CD8^＋ T cells.

Hepatitis B virus (HBV) infection is one of the major global public health problems, with nearly 887 000 deaths from HBV-associated disease each year. Due to the persistent presence of covalently closed circular DNA (cccDNA) in the nuclei of liver cells, patients infected with HBV are prone to chronic infection, which increases the risk of developing cirrhosis and hepatocellular carcinoma. Therefore, closely monitoring cccDNA levels during the treatment of HBV infection plays an important role in assessing the condition of patients with chronic hepatitis B. In contrast, cccDNA testing requires invasive liver puncture to obtain, which is difficult to be widely performed in the clinical work. Existing studies have shown that serum HBV RNA can reflect the levels of cccDNA in patients. This article provides a review of the advances in the application of serum HBV RNA detection during the antiviral treatment process in patients with chronic hepatitis B.

This study constructed a full-length infectious clone of tick-borne encephalitis virus (TBEV), providing an essential tool for in-depth investigation of TBEV replication and pathogenesis. Firstly, prokaryotic promoters within the full-length TBEV sequence were predicted, and synonymous mutations were introduced to eliminate the activity of these promoters, thereby reducing the bacterial toxicity of the cDNA. Subsequently, considering sequence complexity and cloning efficiency, the TBEV cDNA sequence was divided into two fragments for synthesis, which were then cloned into the pFK vector using homologous recombination technology. Viral RNA generated by in vitro transcription was transfected into cells. Viral RNA replication was successfully detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) using TBEV E protein-specific primers. Meanwhile, Western blot analysis with a TBEV NS1 protein-specific antibody confirmed viral protein expression. 50% tissue culture infectious dose (TCID50) assays demonstrated a significant increase in viral titers at 48 h post-transfection. Finally, plaque assays in BHK-21 cells at 96 h post-infection validated the infectivity of second-generation viruses. Results showed that plaque numbers markedly increased with the decreasing viral dilution, and both viral RNA levels and protein expression escalated over time, indicating the infectivity and intracellular replication capacity of second-generation viruses. These results provide a novel tool for TBEV research, and a general strategy for constructing infectious clones.

This study analyzed the clinical characteristics of severe pneumonia in children caused by Whipple disease to enhance pediatricians’ understanding and treatment of this condition. A retrospective analysis was conducted on the clinical data of two cases of severe Whipple disease-related pneumonia admitted to Liuzhou People’s Hospital from 2020 to 2021, along with a review of relevant domestic and international literature. Patient 1 was a female aged 41 months, Patient 2 was a male aged 5 months. Both patients presented primarily with fever, cough, and shortness of breath. C-reactive protein and procalcitonin levels were elevated in both cases, and computed tomography scans indicated varying degrees of patchy shadows and bronchial wall thickening in the lungs. Metagenomic next-generation sequencing was performed on the bronchoalveolar lavage fluid of the patients. In sample 1, Staphylococcus aureus and Tropheryma whipple were detected, while sample 2 revealed Tropheryma whipple and Acinetobacter baumannii. Both patients received assisted ventilation via a ventilator and were treated with a combination of meropenem and sulfamethoxazole-trimethoprim for infection. Both patients eventually improved and were discharged. The results indicate that Whipple disease can cause severe pneumonia in infants and young children, primarily presenting with fever, cough, and shortness of breath, and may be associated with other bacterial infections. It is crucial to employ metagenomic next-generation sequencing technology for pathogen detection early on to confirm the diagnosis and provide timely targeted anti-infection treatment.

Acinetobacter baumannii (AB), a highly pathogenic Gram-negative bacterium, exhibits remarkable environmental adaptability and prolonged survival. It can infect multiple sites in the human body and has become one of the most common pathogens in hospital-acquired infections due to its extensive acquired drug resistance, making the study of its pathogenic mechanisms urgent. Outer membrane proteins (OMPs) are among the key virulence factors of AB, contributing to adhesion and invasion, biofilm formation, immune evasion, and induction of apoptosis. In recent years, with the wide application of advanced technologies such as proteomics, transcriptomics and bioinformatics, significant progress has been obtained in the research on AB OMPs regarding structural biology, vaccine development and target identification, virulence mechanisms, and antimicrobial resistance. This review focuses on the pathogenic mechanisms of AB OMPs and the latest research advancements in this field.

Tuberculous meningitis (TBM) is the most common and severe form of central nervous system tuberculosis, characterized by high mortality and morbidity rates. However, its diagnosis and treatment remain challenging, posing a significant threat to global health. Clinically, the high fatality and disability rates associated with TBM are attributed to multiple factors, including nonspecific clinical manifestations, lack of rapid and sensitive diagnostic tests for early detection, frequent misdiagnosis due to coinfection with other pathogens, the emergence of multidrug-resistant tuberculosis (MDR-TB), and limited blood-brain barrier penetration of antitubercular drugs. This article summarized a case of TBM incidentally diagnosed during COVID-19 screening. A patient presenting with coma underwent cerebrospinal fluid (CSF) smear examination and related laboratory tests. A positive result of acid-fast staining of CSF prompted a rapid clinical diagnosis of TBM. Although molecular diagnostic techniques for TBM demonstrate superior sensitivity and specificity, traditional CSF acid-fast smear microscopy retains unique advantages, such as simplicity, rapid turnaround time and low cost, making it a critical diagnostic tool in resource-limited settings. This study recommends integrating smear microscopy, molecular testing, and culture techniques in clinical practice to establish a stepwise diagnostic framework for TBM.

The envelope proteins of hepatitis B virus (HBV) include large (L), middle (M) and small (S) envelope proteins, with S protein being the most abundant viral transmembrane protein. Functional cure is currently regarded as the therapeutic goal for chronic hepatitis B. One of the key indicators of functional cure is the sustained clearance of circulating hepatitis B surface antigen (HBsAg). However, the mechanisms whereby intracellular S protein is degraded remain elusive. The previous study of this research group demonstrated that G12-CAR, a chimeric antigen receptor (CAR) specifically targeting hepatitis B virus envelope proteins, inhibited S protein secretion and promoted its intracellular degradation. In this study, Huh-7 hepatoma cells transfected by different vectors expressing S protein were treated with bafilomycin A1 and bortezomib, respectively inhibiting lysosomal autophagy and proteasomal degradation pathways. Subsequently, intracellular and extracellular S proteins were detected by Western blot and enzyme-linked immunosorbent assay (ELISA). The results showed that S protein can be degraded via both lysosomal and proteasomal pathways, while other HBV components also can regulate S degradation. Both L protein and 3′-untranslated region (3′-UTR) sequences of the viral S mRNA were identified to affect intracellular S degradation. Hence, these findings may offer novel therapeutic targets for promoting HBsAg clearance.

A total of 82 strains of Klebsiella pneumoniae (KPN) clinically isolated from East Hospital Affiliated to Tongji University in Shanghai from October to December 2022 were collected in this study. Real-time fluorescent polymerase chain reaction (PCR)and enzyme inhibitor enhanced test were used to detect carbapenemase-producing genotypes of the strains. Simultaneously, the minimum inhibitory concentration (MIC) and inhibitory zone diameter of ceftazidime-avibactam (CZA) against these strains of Klebsiella pneumoniae were detected by microbroth dilution method and Kirby-Bauer disk diffusion method, respectively, to verify whether disk diffusion method could serve as a reliable alternative to the microbroth dilution method for CZA susceptibility testing. The results demonstrated complete concordance between carbapenemase genotypes (real-time PCR) and phenotypes (enzyme inhibitor enhancement test). Comparative analysis of the two susceptibility testing methods revealed high agreement between disk diffusion and microbroth dilution. The results of the two disk diffusion methods and the micro-broth dilution method revealed high agreement with categorical agreement (CA) rates of 98.3% and 99.7% and no major errors (ME ＝ 0). This study confirmed that the disk diffusion method can be used to detect the drug susceptibility of CZA to KPN. However, it still needs to be checked by broth dilution method if the inhibition zone range is within 20-22 mm.

Abstract：Predicting the risk of progression to severe/critical coronavirus disease 2019(COVID-19) is crucial for optimizing clinical interventions and healthcare resource allocation. This retrospective study analyzed clinical data from 305 hospitalized COVID-19 patients admitted to Huashan Hospital between December 2022 and January 2023, aiming to identify risk factors and evaluate the predictive performance of a multifactorial model for classifying patients into severe/critical categories. Demographic characteristics, comorbidities, and laboratory parameters (including inflammatory markers, coagulation profiles, and cardiac biomarkers) were collected, with disease severity classified according to the Diagnosis and Treatment Protocol for Novel Coronavirus Infection (Trial Version 10). Multivariable logistic regression revealed that advanced age, decreased lymphocyte count, reduced hemoglobin levels, and elevated levels of C-reactive protein (CRP), interleukin-6 (IL-6), D-dimer, and brain natriuretic peptide (BNP) were independent risk factors for severe/critical classification. A combined predictive model incorporating these indicators achieved an area under the curve (AUC) of 0.748, which significantly outperformed individual predictors (AUC range: 0.625-0.681). The findings suggest that integrating age, inflammatory response, coagulation dysfunction, and cardiac injury biomarkers can effectively identify high-risk COVID-19 patients, providing a scientific foundation for early stratified management, precision treatment, and public health policymaking. This study also contributes evidence-based insights for addressing future emerging infectious disease outbreaks.

The infection and drug sensitivity of Mycoplasma hominis (Mh) and Ureaplasma urealyticum (Uu) in outpatient and inpatient female patients in the Wusong area of Baoshan, Shanghai were studied to provide a scientific evidence for clinical diagnosis and treatment. Mycoplasma culture identification and count drug sensitivity kits were used to carry out Mycoplasma culture, identification and drug susceptible test. A total of 413 cases underwent Mycoplasma culture and drug sensitivity test, of which 178 cases were positive for mycoplasma culture (43.10%), 17 cases with simple Mh infection (4.12%), 152 cases with simple Uu infection (36.80%), and 9 cases with mixed infection (2.18%). In simple Mh infection, the top five high-sensitivity rates from high to low are: doxytetramycin/minocycline, josamycin, clindamycin and gatifloxacin. There is no significant difference in the sensitivity rates of the first three drugs (P>0.05). There is a significant difference between doxytetramycin/minocycline and clindamycin or gatifloxacin (P<0.05, P<0.01). The top five drug resistance rates from low to high are: doxytetramycin/minocycline, josamycin, gatifloxacin, thiamphenicol, and there is no significant difference in the resistance rate of the five drugs (P>0.05). In simple Uu infection, the top five high-sensitivity rates from high to low are: doxytetramycin/minocycline, josamycin, azithromycin and clarithromycin. The difference between the first three drugs has no statistical significance (P>0.05). The difference between doxytetramycin/minocycline and azithromycin or clarithromycin is statistically significant (P<0.01). The drug resistance rates from low to high are: doxytetramycin/minocycline, josamycin, gatifloxacin, azithromycin/erythromycin/levofloxacin, and the resistance rate of doxytetramycin/minocycline is not statistically different from the latter two drugs (P>0.05), and is statistically different from azithromycin/erythromycin/levofloxacin (P<0.05). When treating mycoplasma infections of the reproductive tract, the clinicians should try to choose minocycline, doxytetramycin or josamycin. The alternative drugs for simple Mh and Uu infections are clindamycin and azithromycin respectively.

This study evaluated the diagnostic utility of ultra-multiplex polymerase chain reaction(PCR)-based targeted next-generation sequencing (tNGS) in invasive pulmonary aspergillosis (IPA). A cohort of 209 patients clinically suspected of IPA during May and July 2023 underwent bronchoalveolar lavage fluid (BALF) collection via fiberoptic bronchoscopy. Samples were analyzed using tNGS, galactomannan (GM) assay, and Gram-stained smear. Among 209 suspected IPA cases, tNGS identified Aspergillus in 183 samples, achieving an 87.6% positivity rate. Aspergillus fumigatus was the predominant species (69.4%), followed by Aspergillus flavus (23.5%), Aspergillus terreus (3.83%), and Aspergillus niger (3.3%). GM assay, using a BALF GM index cutoff of ≥1.0, yielded a significantly lower positivity rate of 70.3% (P<0.001), but both outperformed Gram-stained smear (20.1%). Co-detection of other pathogens occurred in 168 tNGS-positive samples (91.8%). Notably, tNGS-derived Aspergillus read counts were significantly higher in GM-positive versus GM-negative patients (P<0.001). Among 15 583 BALF samples analyzed in 2023, tNGS identified Aspergillus in 2 131 (13.7%) samples. Co-pathogens were detected in 91.1% of Aspergillus-positive cases, with influenza virus (25.9%), Klebsiella pneumoniae (14.2%), Acinetobacter baumannii (12.6%), SARS-CoV-2 (12.1%), and Pseudomonas aeruginosa (9.53%) being the predominant other pathogens. This study demonstrates that tNGS surpasses the GM assay in both sensitivity and specificity for IPA diagnosis, enables precise Aspergillus species identification, and excels in detecting fungal co-infections.

This study aimed to develop a new disinfection device for the internal tubing of ventilators and evaluate its efficacy. The tandem device was designed to disinfect the internal tubing of ventilators with hydrogen peroxide (H₂O₂) dry mist that was produced by jet technique. Forty ventilators utilized for mechanical ventilation in confirmed infectious disease cases, each with ≥72 h of continuous operation, were included in the assessment. Colony-forming units (CFU) in air intake, air outlet, and internal tubing, were determined at the following time point, before disinfection, 1 h after disinfection, and 4 h after disinfection. The eligibility criteria were CFU≤20 per sample with no detectable pathogenic bacteria. All data were subjected to statistical analysis. The qualified rate was 0% before disinfection by H2O2, while it was 100% after disinfection for 1 h and 4 h, and CFU in each detection site was significantly lower after disinfection (p＜0.05), and no pathogenic bacterial growth was detected. The tandem H₂O₂ dry mist-disinfecting device is reliable and effective, and the expected effect could be achieved by 1 h disinfection.

The aim of this study was to explore the distribution of infection pathogens and drug resistance of main pathogens in neonates in neonatal intensive care unit (NICU), and to analyze the risk factors of nosocomial infection. A retrospective analysis was performed on the clinical data of 2 812 neonates in NICU of Nanjing Maternal and Child Health Hospital from January 2022 to December 2023. According to the presence or absence of nosocomial infection, neonates were divided into infection group (453 cases) and non-infection group (2 359 cases). The distribution of pathogens and drug resistance of main pathogens in infection group were analyzed. The risk factors of nosocomial infection were analyzed by Logistic regression analysis. Among 2 812 neonates in NICU, there were 453 cases (16.11%) with nosocomial infection. A total of 475 strains of pathogens were detected, mainly Gram-positive bacteria (58.32%). Among Gram-positive bacteria, coagulase-negative staphylococci had high resistance rates to penicillin C (93.57%), erythromycin (76.61%) and oxacillin (71.93%), but were sensitive to rifampin (8.19%), nitrofurantoin (1.75%), vancomycin and linezolid. Staphylococcus aureus was completely resistant to penicillin C (100%), but had no resistance to rifampicin and vancomycin. Among the gram-negative bacteria, Escherichia coli was significantly resistant to ceftriaxone (47.95%) and cefazolin (41.10%), while Klebsiella pneumoniae was highly resistant to ceftazidime (44.64%) and ceftriaxone (39.29%), but both of them were completely sensitive to imipenem and amikacin. Compared with the non-infection group, the infection group had a significantly higher proportion of infants with gestational age <37 weeks, birth weight <2 500 g, cesarean section, Apgar score ≤7 at 5 min after birth, asphyxia rescue, mechanical ventilation, parenteral nutrition, types of antibiotics ≥2, duration of antibiotic use >7 days, and length of ICU stay >7 days (P<0.05). The main pathogens of nosocomial infection are Gram-positive bacteria in NICU neonates, with high resistance rates to penicillin and erythromycin, but still sensitive to vancomycin and linezolid. Gestational age <37 weeks, low birth weight, mechanical ventilation, combined use of antibiotics and stay time in ICU >7 d are independent risk factors of nosocomial infection. It is necessary to standardize invasive operation and antibiotics management, and optimize prevention and control strategies for high-risk neonates.

Salmonella Poona is a rare serotype of Salmonella. This study was conducted on a Salmonella strain isolated from the fecal sample of a patient reported by the active surveillance project of foodborne diseases in Changchun Children’ Hospital. Bacteriological identification, serotyping, drug sensitivity testing, and whole genome sequencing were employed to investigate its biological and molecular profiles. Results demonstrated that this strain was biochemically confirmed as Salmonella and identified as Salmonella Poona by the serological typing. The isolate exhibited resistance to most antibiotics and harbored a diverse array of resistance and virulence genes. This research confirms the first isolation of a multidrug-resistant and highly virulent Salmonella Poona strain in Changchun City, highlighting the need for heightened attention to this pathogen. This study aimed to analyze the biological characteristics and molecular features of Salmonella Poona to provide scientific evidence for clinical treatment and prevention.

This article presented a case of pulmonary infection caused by Actinobacillus ureae (A. ureae) in a patient with malignant tumor, along with a comprehensive literature review. During the patient’s hospitalization for chemotherapy, an opportunistic infection with A. ureae occurred, leading to typical pulmonary inflammation. The laboratory maintained consistent communication with clinicians throughout the diagnosis and treatment process, promptly conducting in vitro antimicrobial susceptibility testing on the pathogen and guiding standardized medication for the patient. Ultimately, following treatment with intravenous cefoperazone-sulbactam combined with levofloxacin for anti-infection symptomatic therapy, the patient’s pulmonary inflammation significantly improved. A literature review revealed that the infection rate of A. ureae was significantly higher than its actual detection level, suggesting that it should be considered one of the potential pathogens in susceptible patients in clinical practice. By reporting this case of pulmonary inflammation caused by an opportunistic infection of A. ureae, this article aims to raise awareness among laboratory staff about this pathogen and provide a reference for clinicians to timely diagnose and treat infections caused by rare pathogens.