Journal: Frontiers in Pain Research

Article Title: The impact of tumor immunogenicity on cancer pain phenotype using syngeneic oral cancer mouse models

doi: 10.3389/fpain.2022.991725

Figure Lengend Snippet: Oral cancer-induced changes in pain-related gene expression. Quantitative PCR was performed to assess changes in Trpv1, Calca , and Atf3 in whole ganglia from (A) sham and MOC1 tumor-bearing mice at PID40 as well as (B) sham and MOC2 tumor-bearing mice at PID14. Gapdh was used as an internal control. Significance was determined by comparing delta CT values between treatment groups within sexes ( n = 5–6/sex/group, Independent t-test, ** p < 0.01). To assess changes in target genes at the single cell level, retrogradely labeled tongue-innervating TG neurons were manually picked to perform single-cell PCR ( n = 36 neurons from 3 mice/sex/treatment). Gene expression was calculated for relative mRNA expression in each cell for each gene. GusB was used as an internal control. The percentage of neurons expressing the target genes was calculated for (C) sham and MOC1 tumor-bearing mice at PID40 as well as (D) sham and MOC2 tumor-bearing mice at PID14. (E–J) Relative mRNA expression values from single-cell PCR for each gene are presented as max to min with all points shown. Expression intensities for each gene between groups was compared using non-parametric Kruskal-Wallis with Bonferroni post hoc test. * p < 0.05, ** p < 0.01.

Article Snippet: Assays from Life Technologies were used to probe for the following genes: Atf3 (Mm00476032_m1), Trpv1 (Mm01246302_m1), Calca (Mm00801463_g1).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control, Labeling, Expressing

Journal: Frontiers in Pain Research

Article Title: The impact of tumor immunogenicity on cancer pain phenotype using syngeneic oral cancer mouse models

doi: 10.3389/fpain.2022.991725

Figure Lengend Snippet: Oral cancer-induced changes in pain-related protein expression. Each TG tissue sections from each retrogradely labeled mouse was stained with antibodies specific for either TRPV1, CGRP or ATF3. Representative immunostaining of all three markers in retrogradely labeled TG tissue from (A) sham and MOC1 mice at PID40 as well as (B) sham and MOC2 mice at PID14. (C,D) The percentage of DiI-labeled neurons expressing each marker was quantified from 2–4 animals per treatment group. Data are presented as mean +/- SEM of percentage of neurons expressing each marker. Two-way ANOVA, Treatment effect: * p < 0.05, ** p < 0.01, sex effect: # p < 0.05.

Article Snippet: Assays from Life Technologies were used to probe for the following genes: Atf3 (Mm00476032_m1), Trpv1 (Mm01246302_m1), Calca (Mm00801463_g1).

Techniques: Expressing, Labeling, Staining, Immunostaining, Marker

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 3. Responsive activation of TRPV1-mediated intracellular Ca2+ concentration. A) CLSM images of 143B and HOS cells in various groups after using the Fluo-4 probe to monitor Ca2+ influx. The scale bar is 20 μm. B,C) FC quantitative value of Ca2+ uptake in 143B and HOS cells after different treatments. D) The protein expression level of TRPV1 was determined by western blot analysis. E) Statistical analyses of TRPV1 (the data are presented as the mean ± SD, n = 3, ***p < 0.001).

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Activation Assay, Concentration Assay, Expressing, Western Blot

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 4. Evaluation of ferroptosis. The production of Lipid-ROS in A) 143B and B) HOS cells after different interventions as measured by CLSM. The scale bars represent 20 μm. C) FC analysis of Lipid-ROS after different interventions in 143B and HOS cells. D,E) Confocal images and FC analyses of ROS generation in 143B and HOS cells treated with different treatments and stained with DCFH-DA (scale bar = 20 μm). F,G) Quantitative value of intracellular C11-BODIPY and DCFH-DA FL intensity (the data are presented as the mean ± SD, n = 3, ***p < 0.001). H) The morphology of mitochondria after various treatments as observed by TEM. The scale bars represent 2 μm. I) The expression level of Gpx4 protein was measured by western blot analysis. J) The expression of TRPV1 and Gpx4 by IF observation after different treatments in 143B cells (scale bar = 100 μm).

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Staining, Expressing, Western Blot

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 5. A) In MG63, HOS, and 143B cells, the effects of CAP on the intracellular oxygen level after different treatments were observed and analyzed by confocal microscopy (scale bar = 20 μm) and FC. B) Quantitative value of intracellular Image-iT Green Hypoxia Reagent FL intensity (the data are presented as the mean ± SD, n = 3, ***p < 0.001). C,D) The expression level of HIF-1𝛼protein was analyzed by western blot (the data are presented as the mean ± SD, n = 3, ***p < 0.001). E,F) The expression of TRPV1 and HIF-1𝛼by IF observation after different treatments in 143B and HOS cells (scale bar = 100 μm).

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Confocal Microscopy, Expressing, Western Blot

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 8. Evaluation and biosafety of different treatments. A) H&E staining and images of PCNA, Ki67, TRPV1, Gpx4, and HIF-1𝛼. IHC staining in tumor specimens after various treatments (scale bar = 100 μm). B–F) IOD values in various groups were analyzed by processing IHC staining through ImagePro Plus. G) Triple IF staining of TRPV1 (red), Gpx4 (orange), and HIF-1𝛼(green) in tumor specimens after various treatments (scale bar = 50 μm). H) TUNEL staining of tumor sections after various treatments (scale bar = 50 μm). I) H&E staining of major organs of mice after different treatments (scale bar = 100 μm). J) Hematological and blood biochemical tests of mice after various treatments.

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Staining, Immunohistochemistry, TUNEL Assay

Journal: Translational Andrology and Urology

Article Title: Evaluation of transurethral GreenLight laser-selective vaporization for refractory overactive bladder in women

doi: 10.21037/tau-24-67

Figure Lengend Snippet: Expressions and comparisons of inflammatory factors and sensory receptors in bladder mucosa tissues before and after treatment. (A) The expressions of TRPV1, P2X3, TNF-α, and IL-6 in patients’ bladder mucosal tissues were stained by immunohistochemical staining before and at 12 weeks after treatment. Representative histological images are shown at 400× magnification. (B) Average optical density value of inflammatory factors and sensory receptors before and after treatment were presented in the bar chart. TNF, tumor necrosis factor; IL-6, interleukin 6; AOD, average optical density.

Article Snippet: Immunohistochemical staining was performed using the avidin-biotin-peroxidase method (ab64212, Abcam, Cambridge, MA, USA) with TRPV1 antibody (CSB-PA822774LA01HU, CUSABIO, Houston, TX, USA), P2X3 antibody (17843-1-AP, Proteintech, Rosemont, IL, USA), tumor necrosis factor alpha (TNF-α) antibody (17590-1-AP, Proteintech, Rosemont, IL, USA), and interleukin 6 (IL-6) antibody (21865-1-AP, Proteintech, Rosemont, IL, USA).

Techniques: Staining, Immunohistochemical staining