Etiological diagnosis value of metagenomic next generation sequencing in the patients with spinal infection / 中华传染病杂志

Objective:To investigate the efficacy of metagenomic next generation sequencing (mNGS) in the etiological diagnosis of patients with spinal infection, so as to provide reference for timely diagnosis and treatment.Methods:A total of 40 patients with suspected spinal infection admitted to the Department of Infectious Diseases in Henan Provincial People′s Hospital from January 2020 to July 2022 were included. The results of tissue culture, histopathological examination and tissue mNGS detection were analyzed retrospectively. According to the clinical diagnose, the patients were divided into the spinal infection group (28 cases) and the non-spinal infection group (12 cases). The positive rate, sensitivity and specificity of mNGS and tissue culture in the pathogen detection of patients with spinal infection were compared. McNemar test was used for statistical analysis.Results:There were 23 males and 17 females in 40 patients. The positive rate of mNGS was higher than that of tissue culture (75.0%(30/40) vs 12.5%(5/40)), and the difference was statistically significant ( χ2=0.08, P<0.001). Based on clinical diagnostic criteria, the sensitivity of mNGS in the diagnosis of spinal infection was higher than that of tissue culture (82.1% vs 17.9%), with a statistically significant difference ( χ2=0.02, P<0.001), while the specificity compared to the tissue culture (33.3% vs 100.0%), the difference was not statistically significant ( P>0.05). Conclusions:mNGS has a high pathogen detection rate and sensitivity in the etiological diagnosis of patients with spinal infection, which could provide clinical guidance for the diagnosis and treatment of patients with spinal infection.

Downregulation of miR-196b in regulating the growth and apoptosis of hepatoma cells by targeting nuclear apoptosis-inducing factor 1 / 临床肝胆病杂志

ObjectiveTo investigate the mechanism of action of miR-196b in regulating the growth and apoptosis of hepatoma cells by targeting nuclear apoptosis-inducing factor 1 (NAIF1). MethodsReal-time PCR was used to measure the expression of miR-196b in hepatoma HuH-7, SNU-449, HepG2, and SMCC7721 cells versus normal human HL7702 hepatocytes. The hepatoma HepG2 cells were collected and divided into Control group (blank control), Anti-NC group (transfected with inhibitor control), Anti-miR-196b group (transfected with miR-196b inhibitor), si-NC group (transfected with siRNA control), si-NAIF1 group (transfected with NAIF1 siRNA), Anti-miR-196b+si-NAIF1 group (co-transfected with miR-196b inhibitor and NAIF1 siRNA), and Anti-miR-196b+si-NC group (co-transfected with miR-196b inhibitor and siRNA control). MTT assay was used to measure the change in proliferation, plate colony formation assay was used to measure colony formation ability, flow cytometry was used to measure cell apoptosis, and Western blot was used to measure the protein expression of Bax and C-caspase-3. Target gene prediction software predicted that NAIF1 might be a target gene of miR-196b, and the luciferase reporting system was used to identify the targeting relationship. The t-test was used for comparison of continuous data between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the SNK-q test was used for further comparison between two groups. ResultsThere was a significant difference in the expression level of miR-196b between hepatoma HuH-7, SNU-449, HepG2, and SMCC7721 cells and normal human HL7702 hepatocytes (1.85±0.16/1.63±012/2.36±0.25/1.92±0.13 vs 1.00±0.09, F=29.05, P＜0.001). Compared with the Anti-NC group, the Anti-miR-196b group had significant reductions in the expression level of miR-196b (0.42±0.03 vs 1.02±0.10, P＜0.05), cell proliferation (0.20±0.02 vs 0.30±0.05, P＜0.05), and colony formation ability (64.35±6.97 vs 119.54±11.82, P＜0.05) and significant increases in apoptosis rate (22.30%±2.09% vs 4.26%±0.35%, P＜0.05) and relative protein expression of Bax (0.69±0.08 vs 0.30±0.05, P＜0.05) and C-caspase-3 (0.63±0.05 vs 0.21±0.04, P＜0.05). Compared with the si-NC group, the si-NAIF1 group had significant increases in proliferation ability (0.46±0.05 vs 0.31±0.04, P＜0.05) and colony formation ability (138.92±9.66 vs 118.47±838, P＜0.05) and significant reductions in apoptosis rate (4.12%±0.40% vs 1.23%±0.12%, P＜0.05), NAIF1 (0.10±0.01 vs 0.17±0.02, P＜0.05), and protein expression of Bax (0.18±0.02 vs 0.29±0.03, P＜0.05) and C-caspase-3 (0.12±0.01 vs 020±0.03, P＜0.05). Compared with the Anti-miR-196b+si-NC group, the Anti-miR-196b+si-NAIF1 group had significant increases in proliferation ability (0.28±0.02 vs 0.21±0.03, P＜0.05) and colony formation ability (97.12±8.23 vs 66.35±5.20, P＜0.05) and significant reductions in apoptosis rate (9.60%±1.11% vs 21.14%±1.32%, P＜0.05), NAIF1 (0.30±0.04 vs 0.52±0.06, P＜0.05), and protein expression of Bax (0.28±0.03 vs 0.67±0.06, P＜0.05) and C-caspase-3 (0.22±0.05 vs 0.60±004, P＜0.05). ConclusionDownregulation of miR-196b can inhibit the growth and induce the apoptosis of hepatoma cells via negative regulation of NAIF1.