Objective To investigate the application value of bronchoalveolar lavage fluid(BALF)met-agenomic next-generation sequencing(mNGS)in elderly patients with severe pneumonia.Methods A total of 48 elderly patients with severe pneumonia admitted to the intensive care unit from January to August 2024 were selected and randomly divided into an observation group and a control group,with 24 patients in each group.BALF samples were collected within 24 hours of ICU admission from both groups,with lavage per-formed at the most radiologically evident lesion site.The control group underwent traditional culture at the hospital,while the observation group had one specimen sent for mNGS testing and another specimen subjected to traditional culture at the hospital.Comparison of detection results between mNGS and traditional culture in the observation group,pathogen distribution,infection markers(on days 4 and 7 of ICU admission),and prog-nostic outcomes were compared between the groups.Results The observation group showed higher positive detection rates and higher rates of detecting ≥2 pathogens by mNGS compared to both traditional culture in the same group and the control group(P＜0.05).Excluding Acinetobacter baumannii,Candida albicans,Can-dida glabrata,and Raoultella planticola,the observation group detected higher quantities of fungi,viruses,and rare pathogens than the control group.On days 4 and 7 of ICU admission,the observation group had signifi-cantly lower body temperature,white blood cell count,C reactive protein,and procalcitonin levels compared to the control group(P＜0.05).The observation group also demonstrated shorter ICU stays,reduced mechanical ventilation duration,and total lower hospitalization costs than the control group(P＜0.05).Conclusion BALF mNGS facilitates early identification of mixed and rare pathogens,improves detection rates,broadens microbial coverage,and shortens testing time.

Background::The calcineurin inhibitor (CNI)-based immune maintenance regimen that is commonly used after renal transplantation has greatly improved early graft survival after transplantation; however, the long-term prognosis of grafts has not been significantly improved. The nephrotoxicity of CNI drugs is one of the main risk factors for the poor long-term prognosis of grafts. Sirolimus (SRL) has been employed as an immunosuppressant in clinical practice for over 20 years and has been found to have no nephrotoxic effects on grafts. Presently, the regimen and timing of SRL application after renal transplantation vary, and clinical data are scarce. Multicenter prospective randomized controlled studies are particularly rare. This study aims to investigate the effects of early conversion to a low-dose CNI combined with SRL on the long-term prognosis of renal transplantation.Methods::Patients who receive four weeks of a standard regimen with CNI + mycophenolic acid (MPA) + glucocorticoid after renal transplantation in multiple transplant centers across China will be included in this study. At week 5, after the operation, patients in the experimental group will receive an additional administration of SRL, a reduction in the CNI drug doses, withdrawal of MPA medication, and maintenance of glucocorticoids. In addition, patients in the control group will receive the maintained standard of care. The patients’ vital signs, routine blood tests, routine urine tests, blood biochemistry, serum creatinine, BK virus (BKV)/cytomegalovirus (CMV), and trough concentrations of CNI drugs and SRL at the baseline and weeks 12, 24, 36, 48, 72, and 104 after conversion will be recorded. Patient survival, graft survival, and estimated glomerular filtration rate will be calculated, and concomitant medications and adverse events will also be recorded.Conclusion::The study data will be utilized to evaluate the efficacy and safety of early conversion to low-dose CNIs combined with SRL in renal transplant patients.Trial registration::Chinese Clinical Trial Registry, ChiCTR1800017277.

Ulcerative Colitis (UC) has been reported to be related to Porphyromonas gingivalis (P. gingivalis). Porphyromonas gingivalis peptidylarginine deiminase (PPAD), a virulence factor released by P. gingivalis, is known to induce inflammatory responses. To explore the pathological relationships between PPAD and UC, we used homologous recombination technology to construct a P. gingivalis strain in which the PPAD gene was deleted (Δppad) and a Δppad strain in which the PPAD gene was restored (comΔppad). C57BL/6 mice were orally gavaged with saline, P. gingivalis, Δppad, or comΔppad twice a week for the entire 40 days (days 0-40), and then, UC was induced by dextran sodium sulfate (DSS) solution for 10 days (days 31-40). P. gingivalis and comΔppad exacerbated DDS-induced colitis, which was determined by assessing the parameters of colon length, disease activity index, and histological activity index, but Δppad failed to exacerbate DDS-induced colitis. Flow cytometry and ELISA revealed that compared with Δppad, P. gingivalis, and comΔppad increased T helper 17 (Th17) cell numbers and interleukin (IL)-17 production but decreased regulatory T cells (Tregs) numbers and IL-10 production in the spleens of mice with UC. We also cocultured P. gingivalis, Δppad, or comΔppad with T lymphocytes in vitro and found that P. gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers. Immunofluorescence staining of colon tissue paraffin sections also confirmed these results. The results suggested that P. gingivalis exacerbated the severity of UC in part via PPAD.
Animals ; Colitis, Ulcerative/microbiology* ; Mice ; Mice, Inbred C57BL ; Porphyromonas gingivalis/pathogenicity* ; Protein-Arginine Deiminases ; Virulence Factors

Objective:To determine the correlations of single nucleotide polymorphisms (SNPs) with atrial fibrillation (AF) in the Chinese Han population from the central plains.Methods:A total of 168 hospitalized patients,including 56 AF and 112 controls,were recruited in this case-control study.The clinical data were obtained from the medical records.All 5 SNPs,rs337711 in KCNN2,rs11264280 near KCNN3,rs17042171 near PITX2,rs6771157 and rs6795970 in SCN10A,were genotyped using amplification refractory mutation system-polymerase chain reaction or direct sequencing.The x2 test was used to compare categorical variables and preliminarily examine correlations between the genotype frequencies and AF.Subsequently,a logistic regression model was constructed to determine the associations between the SNPs and AF based on the above screened results.Odds ratios (ORs) and 95％ confidence interval (CI) were calculated to assess the strength of the correlations.Moreover,we downloaded the genotype data from the HapMap Project for linkage disequilibrium analysis ofrs17042171.Results:AF patients were likely to be of older age and longer left atrial diameter and had more coronary artery disease and higher hypertension compared with the control group (P＜0.05).Among the 5 SNPs,the frequency distribution of genotype AA for rs17042171 was significantly different between the AF and control groups (P＜0.05).After adjusting for several covariates,there was still a high risk ratio in patients with the AA genotype compared with the AC+CC genotype (OR:5.591,95％CI 2.176 to 14.365,P-B＜0.008).Similarly,stratification analysis on the AA genotype demonstrated significant differences between rs17042171 and persistent AF.However,there were not significant correlations between AF and the control groups for the other 4 SNPs (P＜0.05).Conclusion:Rs17042171,near PITX2 on chromosome 4q25,is associated with AF susceptibility in the Chinese Han population from the central plains,suggesting that this SNP can provide a new strategy for clinical diagnosis in AF patients.

BACKGROUND:The in vivo degradation process of chitosan/hydroxyapatite composite porous scaffolds is not very clear. Research on the effects of rat osteoblasts and degradation products is less.
OBJECTIVE:To analyze the biocompatiblity of rat osteoblasts with degradation products of chitosan/hydroxyapatite composite porous scaffolds.
METHODS:The second generation of cultured rat osteoblasts were respectively cultured in the extract of degradation products of chitosan/hydroxyapatite composite scaffolds (experimental group) and Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum (control group). At 2, 4, 6, 8, 10 days of culture, cel counting was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Alkaline phosphatase activity was measured by the recommended method of determination of the Federation, and total protein was determined by BCA method.
RESULTS AND CONCLUSION:The proliferation speed, alkaline phosphatase activity, total cel ular protein synthesis and ratio of alkaline phosphatase to total protein in rat osteoblasts cultured in the experimental group were significantly higher than those in the control group (P<0.05). This experiment showed that the degradation products of chitosan/hydroxyapatite composite porous scaffolds cannot only promote rat osteoblast adhesion, growth and proliferation, but also enhance its ossification function, with good biocompatibility.