Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Concentration-dependent change in the humoral inflammatory response following incubation with Escherichia coli ( E. coli ) in the ex vivo whole blood model. a Absolute plasma concentrations of IL-6, IL-8, and MMP9 determined by enzyme-linked immunosorbent assay. b Normalized values and EC 50 curve fit by BuC=0% and 50 000 CFU/ml E. coli= 100%, respectively, for IL-6, IL-8, and MMP9 as indicated by EC 50 (%) on the respective Y-axis. BuC indicates buffer control after 60 min incubation; numbers on the X-axis indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation; LPS indicates lipopolysaccharide (LPS) 100 ng/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, comparing all shown concentrations of E. coli bacteria and 100 ng/ml LPS with BuC. P -values are indicated above the respective data points. ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Concentration Assay, Incubation, Ex Vivo, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Bacteria

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Diagnostic performance for the detection of bacteremia, analyzing the neutrophil phenotype by determining the median fluorescence intensity (MFI) and the cellular response capacity (CRC) in comparison with traditional markers of humoral inflammation (IL-6, IL-8, MMP9). a Comparison of receiver operating characteristic (ROC) at 10,000 CFU/ml Escherichia coli ( E. coli ) with the respective 95% confidence interval (CI) and P -value, and half-maximal effective concentration (EC 50 ) as a function of the E. coli concentration. b Detailed comparison of the EC 50 as a function of the E. coli concentration. c Exemplary comparison of EC 50 curve fit after normalization as indicated by EC 50 (%) on the respective Y-axis to BuC=100% and 50 000 CFU/ml E. coli =0% for the humoral marker IL-6 (the IL-6 values were multiplied by −1 before EC 50 calculation to facilitate comparability with the CRC) and the change in neutrophil phenotype represented by CD11b CRC. BuC indicates buffer control after 60 min incubation; numbers on the X-axis of c indicate E. coli bacteria in concentrations of 2000 to 50 000 CFU/ml after 60 min incubation. Values are shown as median and interquartile range. n =8. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post-hoc test, evaluating the EC 50 of IL-8, MMP9, the MFI, and CRC of CD10, CD11b, and CD62L in comparison to the EC 50 of IL-6. P -values are indicated above the respective data points. ⁎ P <0.05. CFU. Colony-forming units; IL. Interleukin; MMP9. Matrix metallopeptidase 9.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Diagnostic Assay, Fluorescence, Comparison, Concentration Assay, Marker, Control, Incubation, Bacteria

Journal: Military Medical Research

Article Title: The cellular response capacity (CRC) as a novel immunomonitoring approach in sepsis

doi: 10.1016/j.mmr.2026.100010

Figure Lengend Snippet: Clinical specifications and parameters over all time points of the sepsis cohort. a Suspected infection cause of sepsis. b Distribution of the individual score points of the Sequential Organ Failure Assessment (SOFA) score. c Total SOFA score. d-h Traditional and humoral markers of inflammation: leukocytes and neutrophil-lymphocyte ratio ( d ), C-reactive protein (CRP) and procalcitonin (PCT) ( e ), interleukin-6 (IL-6) and interleukin-8 (IL-8) ( f ), serum amyloid A (SAA) and calprotectin ( g ), matrix metallopeptidase 9 (MMP9) and myeloperoxidase (MPO) ( h ). Values are shown as median and interquartile range. n =14. CNS. Central nervous system; HV. Healthy volunteers.

Article Snippet: For the samples of the ex vivo whole blood model, the plasma concentrations of matrix metallopeptidase 9 (MMP9, #DY911, R&D Systems, Minneapolis, USA), IL-6 (#555220, BD Biosciences, San Jose, USA), and IL-8 (#DY208, R&D Systems) were measured in citrate-anticoagulated plasma using enzyme-linked immunosorbent assay according to the respective manufacturer’s instructions.

Techniques: Infection

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: MMP-9 , a factor that promotes Vasculogenic mimicry, is highly expressed in CC and is associated with poor prognosis. (A) CC database of TCGA was used to analyze key factors associated with VM. (B) Association of Sox2 expression with overall survival in CC (log-rank test). (C) Association of MMP-9 expression with overall survival in CC (log-rank test). (D) Panoramic scans after immunohistochemical detection of MMP-9 and H&E staining in samples from cancerous and paracancerous tissues from subjects with CC. Scale bar, 50 µm. Original magnification, ×20. (E) Protein levels of MMP-9 in 20 paired samples, with the MMP-9 level in CC tissue expressed compared with that in the paired normal tissue. (F) Expression levels of MMP-9 mRNA in 44 paired CC and paracancerous tissues, with MMP-9 expression in CC tissue expressed compared with that in the paired normal tissue. (G) Comparison of the average expression levels of MMP-9 mRNA in CC tissues compared with paracancerous tissues. (H) HeLa and SiHa cells were incubated under hypoxia (0.1% O 2 ) and proteins collected at 24, 48 and 72 h for western blotting of ALDH1, EPHA2, MMP-9 and GAPDH. ImageJ was used to semi-quantify western blotting signals from HeLa (I) and SiHa (J) cells. GAPDH served as an internal reference. *P<0.05, **P<0.01 and ***P<0.001. MMP-9, matrix metalloproteinase 9; VM, vasculogenic mimicry; ALDH1, aldehyde dehydrogenase 1; EPHA2, ephrin type-A receptor 2; TCGA, The Cancer Genome Atlas; Sox2, SRY-box transcription factor 2; CC, cervical cancer; CESC, cervical squamous cell carcinoma.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Immunohistochemical staining, Staining, Comparison, Incubation, Western Blot

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 promotes Vasculogenic mimicry, invasion and migration of CC cells under hypoxic conditions. (A) Expression levels of NSUN2 in CC cells lines (HeLa, SiHa, CaSki, C33A and HT-3) and in a normal cervical cell line (HaCaT). (B) RT-qPCR was used to determine relative expression levels of NSUN2 mRNA in HeLa and SiHa cells after transfection of shRNAs and incubation under hypoxia for 24 h. (C) Relative expression levels of MMP-9 mRNA in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 24 h. (D) Dot blot assay analysis of m 5 C expression levels in HeLa and SiHa cells after 48 h of hypoxia culture following transfection with NSUN2 knockdown plasmids. (E) Semi-quantitative analysis of dot blot results in HeLa cells. (F) Semi-quantitative analysis of dot blot results in SiHa cells. (G) Western blotting was used to investigate NSUN2 and MMP-9 protein levels in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 48 h. ImageJ was used to semi-quantify western blotting bands for (H) NSUN2 and (I) MMP-9 protein levels. (J) 2D tube-forming assays of HeLa and SiHa cells transfected with an NSUN2 -interfering plasmid and incubated under hypoxia. Scale bar, 100 µm. Original magnification, ×4. (K) Assay displayed in panel (J) was quantified using Image Pro. (L) Invasion assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (M) Assay displayed in panel was quantified using Image Pro. (N) Migration assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (O) Assay displayed in panel (N) was quantified using Image Pro. *P<0.05, **P<0.01 and ***P<0.001. VM, Vasculogenic mimicry; CC, cervical cancer; IHC, immunohistochemistry; NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; RT-qPCR, reverse transcription-quantitative PCR; IHC, immunohistochemistry; CaSki, human cervical cancer cell line with intestinal metastasis; C33A, human cervical cancer cell line; HaCaT, human skin keratinocytes cell line; HeLa, human cervical cancer cell line; HT-3, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Migration, Expressing, Quantitative RT-PCR, Transfection, Incubation, Plasmid Preparation, Dot Blot, Knockdown, Quantitative Dot Blot, Western Blot, Invasion Assay, Control, Immunohistochemistry, Reverse Transcription, Real-time Polymerase Chain Reaction, Cloning, shRNA, Negative Control

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 increases the stability of MMP-9 mRNA. (A) A positive correlation was observed between NSUN2 and MMP-9 mRNA expression levels in 44 pairs of samples from subjects with CC. Enrichment of the m 5 C modification of MMP-9 mRNA in HeLa (B) and SiHa (C) Cells were measured with anti-m 5 C methylated-RNA IP assays. Interaction of NSUN2 with MMP-9 mRNA in (D) HeLa and (E) SiHa cells was measured with anti- NSUN2 RNA IP assays. Stability of MMP-9 mRNA after interference with and overexpression of NSUN2 was measured in (F) HeLa and (G) SiHa cells. (H) A model illustrating the proposed mechanism by which NSUN2-mediated stabilization of MMP-9 mRNA promotes Vasculogenic mimicry in CC. NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; HeLa, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; IP, immunoprecipitation; CC, cervical cancer; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Modification, Methylation, Over Expression, Plasmid Preparation, Cloning, shRNA, Immunoprecipitation, Negative Control