Journal: Theranostics

Article Title: Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis

doi: 10.7150/thno.72269

Figure Lengend Snippet: SH2 superbinder exhibited excellent security by targeting lung fibroblasts based on pY levels in vivo. A , Schematic showing progression of SH2 superbinder treatment in control mice. B , Percent survival during last 7 days after SH2 superbinder injection (n = 8). C , HE staining after SH2 superbinder challenge. Images showing the panoramic and partial view of lungs. D , Neutrophil accumulation measured by MPO assay in lung tissues after GST, GST-SH2 WT, GST-SH2 TrM or SH2-TrM treatment in saline groups (n = 6). E , Total cell counting in BALF (n = 6). F-G , Changes in different kinds of cells by Giemsa and Diff-quick staining in BALF (n = 6). H , Inflammatory cytokines in BALF measured by ELISA (n = 6). I , The determination of liver function index in serum (n = 6). J , Schematic showing progression of SH2 superbinder treatment in BLM-treated mice. K , Western blot of GST tag of different tissues in BLM group treated with GST-SH2 TrM. L , The process of FACS for mice lungs. M , Western blot of pY levels in different lung cells which were isolated by FACS of BLM treated 14 days mice. N , The fluorescence images of SH2 superbinder entering into activated fibroblasts, epithelial cells (EpCAM + ), leukocytes (CD45 + ) and endothelial cells (CD31 + ). O-R , Representative images of SH2 superbinder in myofibroblast (α-SMA + ) ( O ), epithelial cells (EpCAM + ) ( P ), leukocytes (CD45 + ) ( Q ) and endothelial cells (CD31 + ) ( R ) of BLM-treated mice.

Article Snippet: The amounts of IL-1β, IL-6, IL-10 and TNF-α in BALF of mice were measured by using mouse ELISA kits from R&D Systems (Minneapolis, MN, USA).

Techniques: In Vivo, Control, Injection, Staining, MPO Assay, Saline, Cell Counting, Diff-Quik, Enzyme-linked Immunosorbent Assay, Western Blot, Isolation, Fluorescence

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: TNFα is correlated with human oral cancer pain scores. ( a ) TNFα protein concentration is higher in cancer tissues compared to anatomically matched contralateral healthy tissues from the same patient (n = 10, * P < 0.05, paired t-test). ( b ) Patients were asked to answer the Oral Cancer Pain Questionnaire before surgery. The mean pain score from patients correlated positively with percentage change in TNFα concentration between cancer and matched contralateral normal tissues ( r = 0.7, P < 0.05).

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Protein Concentration, Concentration Assay

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: Blocking TNFα or JNK inhibits nociception in mice with cancer. ( a ) After 16 weeks of 4NQO treatment, mice exhibited significant increase in gnaw-time from its respective baseline (pre-injection). Propylene glycol (PG) treatment did not affect gnaw-time. In 4NQO tongue cancer mice, C-87 (12.5 mg/kg) IP injection significantly reduced percentage of gnaw-time change from baseline (n = 8) 1 h post-injection than the vehicle (10% DMSO) treated cancer mice (n = 5). C-87 (n = 6) or vehicle (n = 4) had no effect in non-cancer mice treated with PG alone. ( b ) Mice with paw SCC developed cancer pain at PID7. C-87 and the JNK inhibitor SP600125 treatment significantly reduced mechanical nociception compared to vehicle at 1, 3, and 6 h after treatment compared to the control group. 24 h after the treatment the analgesic effect of C-87 was gone (n = 5 per group, Two-way ANOVA). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Blocking Assay, Injection, Control

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: Blocking TNFα inhibits cancer cell growth, migration, and cytokine release. ( a ) Growth rate, measured with the RTCA, following different doses of C-87 treatment in HSC-3 cell culture. C-87 inhibited oral cancer cell growth in a dose dependent manner. One-way ANOVA with Tukey's post hoc analysis. ( b ) Mice with C-87 treatment (n = 7) exhibited a significant decrease in the paw volume compared to the vehicle control mice (n = 6) at PID14, 18, and 21 (two-way ANOVA). Arrow indicates C-87 injection. ( c ) C-87 treated paw cancer mice (n = 6) had smaller tumor area relative to the total paw area compared to vehicle treated paw cancer mice (n = 4). Tumor areas and total paw areas were quantified using H&E stained paw sections. Mann–Whitney U-test. ( d ) Representative H&E stained pictures showing a normal mouse paw, a cancer mouse paw, and a cancer paw treated with C-87 (10 × inset). Scale bar: 100 μm. Images were taken and quantified using Nikon imaging software NIS-Elements F Ver4.60.00. ( e ) C-87 treatment reduced the concentration of TNFα, NGF, IL1β, IL4, MIP3α, IL28β, and IL33 in the paw tumor. Data were presented as fold change of cytokines/chemokines measured from tumor paws over normal paws. n = 6 per group. Mann–Whitney U test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Blocking Assay, Migration, Cell Culture, Control, Injection, Staining, MANN-WHITNEY, Imaging, Software, Concentration Assay

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: TNFα mediates Schwann cell proliferation and migration in vitro. ( a ) The presence of either DOK or HSC-3 cells in cell inserts increased Schwann cell proliferation 48 h following co-culture in the MTS assay. Increased Schwann cell proliferation induced by the presence of HSC-3 cells was inhibited by adding C-87 into inserts. Representative images of cells with Hoechst stain were shown under each culture condition. OD: optical density. ( b ) Schwann cells are more migratory in the presence of HSC-3 cells compared to the DMEM control while DOK reduced Schwann cell migration. Adding C-87 into the HSC-3 culture reduced Schwann cell migration. ( c ) Adding TNFα to the media at the bottom chamber increased Schwann cells migration compared to the DMEM control. Neutralizing TNFα with C-87 decreased Schwann cell migration. ( d ) Schwann cells induced increased HSC-3 cell migration compared to the DMEM control; adding C-87 (20 µM) into the Schwann cell culture in the bottom chamber blocked this increase. ( b – d ), images shown are representative diff-quick stained migrated cells. a-d, one-way ANOVA with Tukey's post hoc analysis. SCs: Schwann cells. Scale bar: 100 μm. * P < 0.05; ** P < 0.01; *** P < 0.001. Images were taken using Nikon imaging software NIS-Elements F Ver4.60.00.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Migration, In Vitro, Co-Culture Assay, MTS Assay, Staining, Control, Cell Culture, Diff-Quik, Imaging, Software

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: The effect TNFα on the expression of Schwann cell activation markers in vitro. TNFα treatment increased c-Jun ( a , b ), GFAP ( c , d ), and p75 ( e – f ) immunofluorescence intensity and protein expression in cultured Schwann cells compared to the DMEM control. TNFα treatment decreased MBP immunofluorescence intensity and protein expression in cultured Schwann cells compared to the DMEM control ( g – h ). Full-length gel blots were provided in the Supplemental Fig. online. Scale bar: 100 μm. Student’s t-test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Expressing, Activation Assay, In Vitro, Immunofluorescence, Cell Culture, Control

Journal: Scientific Reports

Article Title: TNFα promotes oral cancer growth, pain, and Schwann cell activation

doi: 10.1038/s41598-021-81500-4

Figure Lengend Snippet: Activated Schwann cells release increased NGF and TNFα. ( a ) Schwann cells co-cultured with HSC-3 cells overexpressed c-Jun, GFAP, p75 but downregulated MBP compared to control Schwann cells (media alone). Full-length gel blots were provided in Supplemental Fig. online. ( b ) Both DOK and HSC-3 co-cultures increased TNFα mRNA expression in Schwann cells compared to control Schwann cells. ( c ) TNFα protein concentration in Schwann cells co-cultured with either DOK or HSC-3 cells compared to control Schwann cells. ( d ) HSC-3 cell or DRK co-culture increased NGF release in Schwann cells compared with control Schwann cells. ( e ) Adding TNFα in cell culture media stimulated increased NGF release compared with control Schwann cells. One-way ANOVA. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Human NGF and TNFα Quantikine ELISA kits were purchased from R&D systems.

Techniques: Cell Culture, Control, Expressing, Protein Concentration, Co-Culture Assay