Journal: Journal of Advanced Research

Article Title: The pro-cancer and immunosuppressive activity of macrophage-transformed cancer-associated fibroblasts in oral squamous cell carcinoma

doi: 10.1016/j.jare.2025.07.027

Figure Lengend Snippet: MMTs recruit a large number of MDSCs through CXCL10 pathway. A Significantly upregulated genes in MMTs versus non-MMT fibroblasts from OSCC database (GSA-Human: HRA007439). B Differences in CXCL10 expression between non-MMT fibroblasts and MMTs were demonstrated in HNSCC (figure S1). C CXCL10 expression and release were detected by PCR and ELISA. D-H Exogenous CXCL10 was added to BMDMs, while the CXCL10 inhibitor AMG487 was administered to MMTs, followed by co-culturing with MDSCs. The migration of MDSCs was evaluated using transwell chambers and crystal violet staining ( D ). Flow cytometry was used to detect cell apoptosis ( E ), macrophage markers (F4/80 + CD11b + ) ( F ), as well as CFSE for assessing T cell proliferation inhibition ( G ). PCR and ELISA were employed to measure the expression of immunosuppressive factors IL10 and iNOS ( H ). I, J Tumor tissue photographs ( I ) and the tumor growth curve were taken and subjected to statistical analysis regarding volume and weight ( J ). K-M Flow cytometry was used to detect the proportion of MDSCs in the spleen ( K ) and tumors ( L ) of each group, while immunofluorescence staining ( M ) revealed the densest field of MDSCs within the tumor tissues across all groups. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs experimental group. “ns” represents no statistical significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: According to the manufacturer's instructions, the cytokine concentrations of TGFβ1, iNOS, IL-10, and CXCL10 in the supernatant were determined using ELISA kits (Boster, California, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Migration, Staining, Flow Cytometry, Inhibition, Immunofluorescence

Journal: Gut microbes

Article Title: Lacticaseibacillus paracasei sh2020 induced antitumor immunity and synergized with anti-programmed cell death 1 to reduce tumor burden in mice.

doi: 10.1080/19490976.2022.2046246

Figure Lengend Snippet: Figure 7. L. paracasei sh2020 promoted the expression and secretion of CXCL10 in vivo and in vitro. (a) The expression of CXCL9, CXCL10 and CXCL11 in tumor tissues from control and L. paracasei sh2020-treated mice was detected by qRT-PCR. (b-c) Representative images (b) and quantification (c) of IHC staining of CXCL10 in the tumor tissues from control and L. paracasei sh2020-treated tumors. (d) Tumor growth in each group. (e) The levels of CXCL10 in the conditioned medium. (f-g) Tumor growth in the tumor-bearing mice with intratumoral injection of L. paracasei sh2020 (n = 5–6). (h-i) Representative images (h) and quantification (i) of IHC staining of CXCL10 and CD8 in each group (n = 4–5). (j) The serum levels of CXCL10 were examined by ELISA. ns, no significant difference, *P < .05, **P < .01, ***P < .001.

Article Snippet: Depletion of T cell subsets and CXCL10 in vivo In the in vivo T cell subsets depletion study, InVivoMAb anti-mouse CD4 (YTS191, BE0119, BioXCell), CD8α (YTS169.4, BE0117 BioXcell) antibodies were intraperitoneally injected at a dose of 200 μg per mouse.

Techniques: Expressing, In Vivo, In Vitro, Control, Quantitative RT-PCR, Immunohistochemistry, Injection, Enzyme-linked Immunosorbent Assay

Journal: Gut microbes

Article Title: Lacticaseibacillus paracasei sh2020 induced antitumor immunity and synergized with anti-programmed cell death 1 to reduce tumor burden in mice.

doi: 10.1080/19490976.2022.2046246

Figure Lengend Snippet: Figure 8. CXCL10 controlled CD8+ T cell migration and the effect of L. paracasei sh2020 in vivo. (a-b) Representative images of IHC staining of CD8 and CXCL10 (a), and quantification (b) for the control and L. paracasei sh2020-treated tumors (n = 6–7). (c) IHC analysis of CD8 in tumors, which were divided into two groups according to CXCL10 high and low expression. (d) Experimental design: C57BL/6 mice were implanted subcutaneously with 5.0 × 105 MC38 cells and was treated with control vehicle or anti-CXCL10 antibody by intraperitoneal injection, every 3 days starting on D3, in total three times. The mice were given L. paracasei sh2020 with a dose of 1.0 × 109 CFU by gavage starting from D0 to D13. (e) Tumor growth in tumor-bearing mice in d. (f) Quantification of IHC staining of CXCL10 and CD8 in the tumors after neutralizing CXCL10 in vivo (n = 4–5). ns, no significant difference, *P < .05, **P < .01, ***P < .001.

Article Snippet: Depletion of T cell subsets and CXCL10 in vivo In the in vivo T cell subsets depletion study, InVivoMAb anti-mouse CD4 (YTS191, BE0119, BioXCell), CD8α (YTS169.4, BE0117 BioXcell) antibodies were intraperitoneally injected at a dose of 200 μg per mouse.

Techniques: Migration, In Vivo, Immunohistochemistry, Control, Expressing, Injection

Journal: Medicine

Article Title: Determination of diagnostic and predictive parameters for vertical mandibular invasion in patients with lower gingival squamous cell carcinoma: A retrospective study

doi: 10.1097/MD.0000000000032206

Figure Lengend Snippet: Immunohistochemical staining.

Article Snippet: IL-6 , Rockland Inc. , 1:2000 , Rabbit polyclonal antibody , • Inflammatory cytokines released by cancer cells. • Since stimulation by IL-6 and TNF-α induces bone-resorbing multinucleated giant cells, it is possible that some mechanisms of the bone-resorptive effects of IL-6 are not mediated by the RANK/RANKL pathway..

Techniques: Immunohistochemical staining, Staining, Binding Assay, Activity Assay, Expressing

Journal: EMBO Molecular Medicine

Article Title: Adaptively evolved human oral actinomyces‐sourced defensins show therapeutic potential

doi: 10.15252/emmm.202114499

Figure Lengend Snippet: An inhibition‐zone‐based assay for evaluating the stability of AMSIN in H 2 O or normal mouse serum. The bacterium used was BM and the peptide dose was 0.2 nmol/well. Mean ± SD of three biological replicates is displayed. P values were obtained by Mann–Whitney U ‐test (* P < 0.05, ns: no significance; exact P values are listed in Table ). Hemolysis by AMSIN. Meucin‐18 was used as a positive control. Mean ± SD of three technical replicates is displayed. Comparison of cytotoxic effects of AMSIN and LL‐37 on HL‐60 cells. Mean ± SD of three biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by a red asterisk; * P < 0.05, *** P < 0.001, ns: no significance; exact P values are listed in Table ). An inhibition‐zone assay showing the inability of AMSIN to bind DNA. The bacterium used was MRSA P1374 and the peptide dose was 1.0 nmol/well. Mean ± SD of three biological replicates is displayed. P values were obtained by Mann–Whitney U ‐test (ns: no significance; exact P values are listed in Table ). The effect of AMSIN on human ion channels expressed in the CNS and heart. The asterisks mark steady‐state current traces after administering 100 μM AMSIN. Peptide‐induced IL‐8 release. A549 cells were treated with AMSIN or LL‐37 for 24 h. IL‐8 levels in culture supernatants from the FBS‐containing medium ( left ) or the serum‐free medium ( right ) were measured with an ELISA, as described in Materials and Methods. Mean ± SD of three biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by a red asterisk or ns; * P < 0.05, ** P < 0.01, *** P < 0.001, ns: no significance; exact P values are listed in Table ). Peptide‐induced MCP‐1 release. HL‐60 cells were treated with AMSIN or LL‐37 for 24 h. MCP‐1 levels in culture supernatants from the FBS‐containing medium ( left ) or the serum‐free medium ( right ) were measured with an ELISA. Mean ± SD of three biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by a red asterisk or ns; * P < 0.05, ** P < 0.01, ns: no significance; exact P values are listed in Table ). Surface plasmon resonance (SPR)‐based assay detecting anti‐AMSIN antibodies in mouse serum using Biacore T100. Sera were taken from three AMSIN‐treated mice (Samples 1–3) and one control mouse only injected with 0.9% NaCl. The sensorgrams present data of subtracting the background SPR signals from the control. Note: the abnormal peaks at 30 and 90 s yielded by the equipment due to sample injections were artificially removed. Insect, schematic diagram of SPR experiment in which 1:100 diluted serum was used as analyte to flow over the AMSIN‐immobilized CM5 chip. Treatment of pneumococcal pneumonia. Five mice per sampling point were inoculated with SP D39 through the nasopharynx and 24 h later the animals received AMSIN or penicillin (as a positive control) at different doses (AMSIN, at the doses of 8.35, 16.7, and 33 mg per kg; penicillin at 8.35, 16.7, and 30 mg per kg) via intramuscular injection (i.m.). One day after treatment, the animals were necropsied and their lungs were removed and homogenized for the determination of the level of SP , as described in the Materials and Methods section. Each point represents the determination from a single animal and the line shows the mean log value. Mean ± SD of five to ten biological replicates is displayed. P values were obtained by Student’s t ‐test or Mann–Whitney U ‐test (the latter indicated by red asterisks; ** P < 0.01, *** P < 0.001 denotes significant reduction in CFU compared with the saline group; exact P values are listed in Table ). Survival after peritoneal infection with MRSA P1374. The survival fraction of the vehicle‐treated control group was zero out of 11 mice and the treatment group contained six mice and all survived. Source data are available online for this figure.

Article Snippet: Medium was aspirated and replaced with fresh complete DMEM supplemented with 10% FBS or serum‐free medium, both containing different concentrations of polypeptides for 24 h. Then, the supernatants were harvested for quantification of the IL‐8 amount in A549 by the Human CXCL8/IL‐8 Quantikine ELISA Kit (R&D Systems Inc., Minneapolis, MN, USA) and the MCP‐1 amount in HL‐60 by the Human MCP‐1 ELISA Kit (Boster Biological Technology Co., Ltd, Wuhan, China) as per the manufactures’ instructions.

Techniques: Inhibition, MANN-WHITNEY, Positive Control, Comparison, Enzyme-linked Immunosorbent Assay, SPR Assay, Control, Injection, Sampling, Saline, Infection