Original Article Metabonomic analysis of cerebrospinal fluid in epilepsy Di Niu, Pin Sun, Fenghua Zhang, Fan Song

Objective. This study retrospectively analyzed the clinical diagnosis, treatment process, and laboratory test data of patients with pulmonary cryptococcosis to improve the understanding and diagnosis and treatment ability of the disease. Methods. Patients with pulmonary cryptococcosis diagnosed in the First Affiliated Hospital of Dalian Medical University from October 2003 to July 2021 were selected, and their medical records were consulted. The general data, clinical manifestations, laboratory examinations, imaging characteristics, diagnosis, and treatment methods were studied. The software SPSS 22 was used for statistical analysis. Results. A total of 50 patients with pulmonary cryptococcosis were included in the study. The ratio of male to female was 1 : 1. The average age was years with a range of 27-82 years. Grouping the patients by age, with 10 years as an age group, we found that 40-60 years was the high-incidence age group. Two patients (4%) had a history of bird contact, and 18 patients (36%) had at least one underlying conditions. Hypertension and cough were the most common underlying condition and clinical manifestation, respectively. The main admission diagnoses were lung shadow (19/50, 38%) and chest/lung mass (15/50, 30%). In the imaging findings, the most common type of lesions was nodule/nodule shadow (29/69, 42.03%). Lesion distribution in the lower lobe, single lobe, and right lung was more frequent than that in the upper lobe, multilobes, and left lung, respectively. Burr sign (12/43, 27.91%) was the most common concomitant sign. Pulmonary ventilatory defect was found in 7 cases. Laboratory test results were largely nonspecific. The pathological examination showed granuloma, with 47 cases (94%) confirmed by postoperative biopsy. Two cases (4%) were confirmed by serology. One case (2%) was diagnosed with Cryptococcus smear. 43 cases (86%) were treated with simple surgical resection, 6 cases (12%) were treated with antifungal drugs, and 1 case (2%) was transferred to another hospital for suspicion of pulmonary tuberculosis. Conclusions. Pulmonary cryptococcosis is more common in the middle-aged and elderly, and the clinical specificity is low. It can occur in people with normal or impaired immune function. The main clinical and imaging manifestation is cough and pulmonary nodules, which are very easy 5to be misdiagnosed. Surgical resection is the primary treatment.

Objective: In this study, Cryptococcus neoformans BNCC225501 was used as research objects to explore the antifungal activity of magnolol. The molecular mechanisms underlying magnolol against Cryptococcus neoformans BNCC225501 were explored using metabolomics and transcriptomics. The results offer ideas and directions for developing new antifungal drugs based on magnolol. Method: The minimum inhibitory concentration (MIC) and the method of drawing the growth curve were used to determine whether magnolol had fungistatic and fungicidal effects on Cryptococcus neoformans . Cryptococcus neoformans was cultured on urea basal medium and Niger seed medium, and different concentrations of magnolol were added to these media to detect the effects of magnolol on the urease and melanin of Cryptococcus neoformans . Network pharmacology was used to screen for potential target genes of magnolol and fungal infection, obtaining common targets. Further molecular docking was performed between magnolol and the proteins corresponding to the common targets. Results: The MIC of the individual tested agents against Cryptococcus neoformans strain was 8 μg/mL for magnolol. The combination of magnolol and fluconazole revealed good synergistic effects against Cryptococcus neoformans (FIC ≤ 0.5). At a magnolol concentration of 8 μg/mL, the expression of urease was inhibited. Compared to the control group, in the experimental group treated with magnolol, among the differential metabolites, lipids and lipid‐like molecules accounted for the highest percentage of differential metabolites, except for unclassified metabolites. A total of 30% of the metabolites belonged to lipids and lipid‐like molecules, followed by amino acids, peptides, and analogues (22.63%), organoheterocyclic compounds (7.91%), alkaloids (6.17%), sugars (6.01%), nucleosides, nucleotides and analogues (3.80%), organic oxygen compounds (2.22%), aromatic compounds (1.90%), organic acids (1.90%), and other categories (16.93%). KEGG pathway enrichment analysis results showed that the metabolites were mainly enriched in glyoxylate and dicarboxylate metabolism, arginine biosynthesis, valine, leucine, and isoleucine degradation; arginine and proline metabolism; cysteine and methionine metabolism; pentose and glucuronate interconversions; sphingolipid metabolism; histidine metabolism; and the citrate cycle. Compared to the control group, in the experimental group treated with magnolol, a total of 928 genes were detected to be altered. The GO enrichment results showed that the altered genes were enriched in the component of membrane and oxidation‐reduction process. KEGG enrichment analysis showed that the differential genes were mainly enriched in the DNA replication, NOD‐like receptor signaling pathway, and fatty acid biosynthesis. Combined analysis of metabolomics and transcriptomics demonstrated that the glycerophospholipid metabolism pathway and glutathione metabolism pathway were significantly changed after treatment with magnolol. Screening identified 31 target genes for magnolol and 969 fungal genes, with three intersecting targets, DPP4, MAPK14, and CCNA2. Molecular docking studies showed that magnolol and the target protein had a good binding ability, a docking binding energy of −7.3 kcal/mol. Conclusions: Magnolol has fungistatic and fungicidal ability for Cryptococcus neoformans and can inhibit the urease production capacity of Cryptococcus neoformans . The antifungal mechanism of magnolol may be that it disrupts redox homeostasis and inhibits detoxification by affecting Glutathione metabolism.

Background: In recent years, the outbreak of COVID-19 caused by SARS-CoV-2 has been witnessed globally. However, the impact of SARS-CoV-2 infection on thyroid dysfunction and subclinical thyroid dysfunction remains unclear. Therefore, this meta-analysis aimed to assess the effects of SARS-CoV-2 infection on thyroid dysfunction and its relationship with the severity of COVID-19.Methods: We systematically searched databases including PubMed, Willey Library, Embase, Web of Science, CNKI, Wanfang, and VIP. We focused on randomized controlled trials, case-control studies, and cohort studies published between December 2019 and August 2023, examining the association between SARS-CoV-2 infection and hypothyroidism, with a specific emphasis on the severity of the infection. The quality of the research was assessed using the Newcastle-Ottawa Scale (NOS), while statistical analysis was conducted using the meta and metafor packages in R 4.2.1 software.Results: For the meta-analysis, a total of eight articles were identified based on strict inclusion and exclusion criteria. For the association between SARS-CoV-2 infection and hypothyroidism, three studies (266 samples) comparing TSH levels of COVID-19 and control groups showed no difference in TSH levels [SMD=-0.04,95%CI(-1.22,1.15),P=0.95]. Additionally, two studies examining TT3 (a sample of 176 cases) and two studies examining TT4 (a sample of 176 cases) also showed no difference in TT3 and TT4 between the COVID-19 group and the control group, respectively. However, when evaluating the severity of COVID-19, six studies (565 samples) showed that TSH in the severe group was significantly lower than in the mild group [SMD = -0.55, 95% CI (-0.96, -0.14)], while FT3 was also lower in the severe group [SMD = -0.96, 95% CI (-1.24, -0.67)]. No noticeable differences were observed between the severe and mild groups in their TT3, FT4, and TT4 levels.Conclusion: SARS-CoV-2 infection may have detrimental effects on thyroid function in individuals with severe symptoms. More research is needed to confirm and explore this relationship.Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO, identifier CRD42023486042.

Introduction: Patients with chronic kidney disease (CKD) are at high risk of morbidity and mortality from SARS-CoV-2 infection (COVID-19). However, their immune response to vaccination may vary among individuals. The purpose of this review was to identify characteristics of alterations in humoral and cellular immune responses to the vaccination, and to provide insights into their immune dysfunctions for a better care of acute COVID-19 and prevention of long COVID-19. Methods: PubMed, Embase, Scopus, Web of science and Cochrane Central were systematically searched. Eligible publications included clinical studies reporting immune response to COVID-19 vaccination in CKD patients without dialysis or KT, CKD patients undergoing dialysis, as well as CKD patients with KT. Demographics, measurements and results of their humoral and cellular response were evaluated, and the quality of studies were assessed using the Joanna Briggs Institute (JBI) critical appraisal tool and the Newcastle-Ottawa quality assessment scale (NOS). Results: A total of 31 eligible studies were identified. A decreased proportion of patients with KT showed anti-S IgG positivity after the 2 ^nd (67%) and 3 ^rd (56.6%) dose of vaccination. Similarly, a decreased proportion of these patients presented S-specific T-cell response after the 2 ^nd (17.7%) and 3 ^rd (12.9%) dose. Though lower anti-S IgG titers in patients with CKD or on dialysis, as well as T-cell response in patients on dialysis were reported to be lower after the 2 ^nd or 3 ^rd dose of vaccination, conflicting results were reported by other studies. Limited studies on correlated change between humoral and cellular immune response revealed a low rate of co-presence of the two in patients with dialysis, though antibody level was correlated with rate of cellular response, while no such correlation was revealed in patients with KT. Conclusion: The study provides crucial information on features of humoral and cellular immune responses to COVID-19 vaccinations in CKD patients, and suggests possible directions for strategy of management such as antibody monitoring, additional booster dose or immunomodulatory therapies not only for acute COVID-19 but also for long COVID-19.

Objective: To analyze the phenotypic and genomic characteristics of a non-typhoidal Salmonella bile isolate from a patient with chronic diarrhoea secondary to acute cholecystitis. Methods: The patient presented with chronic diarrhoea lasting for two weeks, which was secondary to acute cholecystitis. The non-typhoidal Salmonella bile isolate was identified using biochemical tests, mass spectrometry, serum agglutination tests, and antimicrobial susceptibility testing. Subsequently, whole-genome sequencing (WGS) was performed to predict and annotate the serotype, multilocus sequence typing (MLST), antimicrobial resistance genes (AMR), mobile genetic elements (MGEs), and virulence genes of the isolate. Results: The isolate was identified as Salmonella spp. by biochemical tests and mass spectrometry. The serotype of the isolate identified by serum agglutination tests was consistent with WGS prediction and was identified as Salmonella Stanley (1,4,5,12: d:1,2, ST29). It was resistant to six antimicrobial agents, including ampicillin, ciprofloxacin, cefotaxime, azithromycin, trimethoprim/sulfamethoxazole, and chloramphenicol. Five major classes of antimicrobial agents, comprising a total of 14 resistance genes were screened, including β-lactam resistance genes: bla_TEM-1B, bla_OXA-1, and bla_DHA-1; quinolone resistance gene: qnrB4 and aac(6′)-Ib-cr; macrolide resistance genes: mph(A), mph(E), and msr(E), and folate pathway inhibitor resistance genes: sul1, sul2, and dfrA14; chloramphenicol resistance genes: floR, catA2 and catB3. T57S mutation in the parC gene was detected in the quinolone resistance-determining region (QRDR). Three plasmid replicons (IncHI2/IncHI2A, IncR, IncN) and three insertion MGEs were predicted. Forty-two virulence genes were predicted, of which 25 were secretion and transporter genes, and ten were fimbrial adherence genes. Only one invasive protein-regulated gene(inv) was found in Salmonella Pathogenicity Islands 1(SPI-1), and no Typhoid toxin genes were predicted. Conclusion: The case of Salmonella Stanley ST29 isolated from a patient with diarrhoea lasting 55 days secondary to cholecystitis aligns with the characteristics of high drug resistance and relatively low virulence. This highlights the need for increased vigilance in clinical practice regarding invasive cases and the significant disease burden associated with the intestinal migration of antimicrobial-resistant Salmonella isolates.