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.

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.

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 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 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.

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.

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.

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.