Journal: Investigative and Clinical Urology

Article Title: Improving the diagnosis of high grade and stage bladder cancer by detecting increased urinary calprotectin expression in tumor tissue and tumor-associated inflammatory response

doi: 10.4111/icu.2019.60.5.343

Figure Lengend Snippet: Comparison of urinary and tissue calprotectin levels of patient and control groups

Article Snippet: The level of urinary calprotectin was measured by the enzyme-linked immunosorbent assay (ELISA) method (Human Calprotectin ELISA kit; MyBiosource, San Diego, CA, USA) after centrifuging the urine samples at 3,000 rpm for 20 minutes.

Techniques: Comparison, Control

Journal: Investigative and Clinical Urology

Article Title: Improving the diagnosis of high grade and stage bladder cancer by detecting increased urinary calprotectin expression in tumor tissue and tumor-associated inflammatory response

doi: 10.4111/icu.2019.60.5.343

Figure Lengend Snippet: Comparison of urinary and tissue calprotectin levels of patient and control groups (female patients excluded)

Article Snippet: The level of urinary calprotectin was measured by the enzyme-linked immunosorbent assay (ELISA) method (Human Calprotectin ELISA kit; MyBiosource, San Diego, CA, USA) after centrifuging the urine samples at 3,000 rpm for 20 minutes.

Techniques: Comparison, Control

Journal: Investigative and Clinical Urology

Article Title: Improving the diagnosis of high grade and stage bladder cancer by detecting increased urinary calprotectin expression in tumor tissue and tumor-associated inflammatory response

doi: 10.4111/icu.2019.60.5.343

Figure Lengend Snippet: Urinary and tissue calprotectin levels of papillary, solid, and mixed tumors

Article Snippet: The level of urinary calprotectin was measured by the enzyme-linked immunosorbent assay (ELISA) method (Human Calprotectin ELISA kit; MyBiosource, San Diego, CA, USA) after centrifuging the urine samples at 3,000 rpm for 20 minutes.

Techniques:

Journal: Investigative and Clinical Urology

Article Title: Improving the diagnosis of high grade and stage bladder cancer by detecting increased urinary calprotectin expression in tumor tissue and tumor-associated inflammatory response

doi: 10.4111/icu.2019.60.5.343

Figure Lengend Snippet: Comparison of tumor grade and urinary and tissue calprotectin levels

Article Snippet: The level of urinary calprotectin was measured by the enzyme-linked immunosorbent assay (ELISA) method (Human Calprotectin ELISA kit; MyBiosource, San Diego, CA, USA) after centrifuging the urine samples at 3,000 rpm for 20 minutes.

Techniques: Comparison

Journal: Investigative and Clinical Urology

Article Title: Improving the diagnosis of high grade and stage bladder cancer by detecting increased urinary calprotectin expression in tumor tissue and tumor-associated inflammatory response

doi: 10.4111/icu.2019.60.5.343

Figure Lengend Snippet: Comparison of tumor stage with urinary and tissue calprotectin levels

Article Snippet: The level of urinary calprotectin was measured by the enzyme-linked immunosorbent assay (ELISA) method (Human Calprotectin ELISA kit; MyBiosource, San Diego, CA, USA) after centrifuging the urine samples at 3,000 rpm for 20 minutes.

Techniques: Comparison

Journal: Investigative and Clinical Urology

Article Title: Improving the diagnosis of high grade and stage bladder cancer by detecting increased urinary calprotectin expression in tumor tissue and tumor-associated inflammatory response

doi: 10.4111/icu.2019.60.5.343

Figure Lengend Snippet: Receiver operating characteristic (ROC) curve analysis of urinary and tissue calprotectin levels in the diagnosis of primary bladder cancer (area under the curve [AUC]=0.976, 95% confidence interval [CI], 0.929–1.023, p<0.0001; AUC=0.713, 95% CI, 0.579–0.847; respectively, for urinary and tissue calprotectin levels).

Article Snippet: The level of urinary calprotectin was measured by the enzyme-linked immunosorbent assay (ELISA) method (Human Calprotectin ELISA kit; MyBiosource, San Diego, CA, USA) after centrifuging the urine samples at 3,000 rpm for 20 minutes.

Techniques: Biomarker Discovery

Journal: Investigative and Clinical Urology

Article Title: Improving the diagnosis of high grade and stage bladder cancer by detecting increased urinary calprotectin expression in tumor tissue and tumor-associated inflammatory response

doi: 10.4111/icu.2019.60.5.343

Figure Lengend Snippet: Receiver operating characteristic (ROC) curve analyses of urinary calprotectin level in the discrimination of high-grade from lower grade tumors and non-muscle-invasive from muscle-invasive tumors (area under the curve [AUC]=0.92, 95% confidence interval [CI], 0.671–0.913, p<0.0001; and AUC=0.85, 95% CI, 0.69–1.00, p=0.01; respectively).

Article Snippet: The level of urinary calprotectin was measured by the enzyme-linked immunosorbent assay (ELISA) method (Human Calprotectin ELISA kit; MyBiosource, San Diego, CA, USA) after centrifuging the urine samples at 3,000 rpm for 20 minutes.

Techniques:

Journal: Journal of cancer science & therapy

Article Title: Blocking PD1/PDL1 Interactions Together with MLN4924 Therapy is a Potential Strategy for Glioma Treatment.

doi: 10.4172/1948-5956.1000543

Figure Lengend Snippet: Figure 1: PDL1 expression in clinical brain tumor samples, glioma cell lines and CSF samples from the immunocompetent glioma mice model. A) Graph (left) represents PDL1 mRNA expression in brain tumors and melanomas (cBioPortal (http://www.cbioportal.org)). Note that the average PDL1 mRNA level in GBM is similar to the average PDL1 mRNA level in melanomas. The PDL1 values in GBM and low-grade gliomas which exceed the average PDL1 mRNA level in melanomas are highlighted in the red box. Kaplan-Meier plot (right) illustrates survival rates of glioma patients with high and low PDL1 expression levels; 9% versus 24% of 2 year survival for high and low PDL1 expression; the difference is significant, P=0.02 (data has been obtained from the Human Pathology Atlas). B) Immunohistochemical detection of PDL1 in the tissue microarray of normal and brain tumor samples. The images were taken at 40x magnification. C) Western blot illustrates PDL1 and Actin protein levels in control and brain tumor clinical samples. Graph represents PDL1 to Actin ratios for corresponding protein samples; 0.87 ± 0.32 (n=7) versus 0.19 ± 0.1 (n=7) for tumor and normal samples, respectively, the difference is significant, P=0.003. Two samples (marked by light grey color) of patients with hemorrhage have been excluded from the average. D) Western blots illustrate PDL1 protein levels in established and PDGx glioma cell lines (left) and in the extracellular media collected from these cell lines (right). The protein content of the collected media was concentrated six folds before an analysis (see method). The graph (right) provides PDL1 concentrations in CSF samples from control and tumor-bearing mice (see method). The immunocompetent glioma mice model with GL261 cells was utilized for this experiment. E) Images illustrate interactions of tumor neuro spheres from XD456 (a) and from two parental U251-IDH1-R132H cell lines with different PDL1 expression levels (b) with primary T-cells. Note that tumor neuro spheres with high PDL1 levels keep their integrity after 48 hours of interaction with primary T-cells (illustrated in the insert). The graph represents the average percent of neuro spheres after 48 hours of interaction with T-cells, 89 ± 7% (n=4) and 14 ± 5% (n=4) for cell lines with high and low PDL1 levels, respectively. The difference is significant, P=0.0002. Primary T-cells were loaded with calcein, AM (green) before experiments for visualization.

Article Snippet: The PDL1 level in CSF samples was analyzed by using the Mouse PDL1 ELISA Kit (Boster, Pleasanton, CA, USA).

Techniques: Expressing, Immunohistochemical staining, Microarray, Western Blot, Control

Journal: Journal of cancer science & therapy

Article Title: Blocking PD1/PDL1 Interactions Together with MLN4924 Therapy is a Potential Strategy for Glioma Treatment.

doi: 10.4172/1948-5956.1000543

Figure Lengend Snippet: Figure 2: HIF1A accumulation evokes PDL1 up-regulation in glioma cell lines. A) HIF1A accumulations induced by CoCl2 treatment of U251 and U87 cell lines evoke PDL1 up-regulation in protein and mRNA levels. Western blot illustrates HIF1A and PDL1 levels in nuclear and cytoplasmic fractions, respectively, in control and after treatment with CoCl2 (85uM). Lamin A/C and alpha Tubulin were utilized to verify nuclear and cytoplasmic fractions, respectively. Graphs illustrate normalized PDL1/ GAPDH mRNA ratios after cell treatment with CoCl2 (85uM) at different time points. In each experiment, data has been normalized to the corresponding ratios in untreated cells, results are presented as mean ± S.D. B) Transcription initiation complexes encoded by pAC154-dual-dCas9VP160 plasmids, guided by sgRNAs to the HIF1A binding domains in the first intron of the PDL1 gene, evoke PDL1 overexpression. Western blot illustrates PDL1 and Actin protein levels in U251 cells after transfection with plasmids encoding control (scrambled sequence) sgRNA or HIF1A sgRNAs. Note that transcription initiation complexes guided by sgRNAs to the HIF1A binding domains (A), (B), and (A) with (B) simultaneously increased PDL1 expression by 1.5, 3.9 and 1.8 folds, respectively, compared to the control (PDL1 expression in the presence of transcription initiation complexes guided by scrambled sgRNA). In each experiment, PDL1 expression was normalized to the Actin expression.

Article Snippet: The PDL1 level in CSF samples was analyzed by using the Mouse PDL1 ELISA Kit (Boster, Pleasanton, CA, USA).

Techniques: Western Blot, Control, Binding Assay, Over Expression, Transfection, Sequencing, Expressing

Journal: Journal of cancer science & therapy

Article Title: Blocking PD1/PDL1 Interactions Together with MLN4924 Therapy is a Potential Strategy for Glioma Treatment.

doi: 10.4172/1948-5956.1000543

Figure Lengend Snippet: Figure 3: MLN4924 treatment induces up-regulation of HIF1A and PDL1 in glioma cell lines. A) The inhibitory dose- response curves for MLN4924 in established, PDGx and PDGX-stem glioma cell lines. The IC50s are 0.3 ± 0.2 uM (n=4), 2.7 ± 1 uM (n=6), 3 ± 2 uM (n=3), 3 ± 1 uM (n=4), 2.9 ± 0.5 uM (n=4), 0.8 ± 0.2 uM (n =4), 0.2 ± 0.1uM (n=4) for LN221, U251, U87, XD456, JX10, XD456-stem, X14P-stem cell lines, respectively, after treatment with MLN4924 for 5 days. B) Western blots illustrate HIF1A and PDL1 protein levels in nuclear and cytoplasmic fractions in the control and after treatment with MLN4924 (1 uM, 5 days). LaminA/C and alpha Tubulin were utilized to verify nuclear and cytoplasmic fractions, respectively. C) The graph illustrates normalized PDL1/18S mRNA ratios after treatment with MLN4924 (1uM, 5 days) for different cell lines. Note the significant enhancement of the PDL1/18 mRNA ratio for all cell lines after MLN4924 treatment: 8 ± 3, 25 ± 5, 5 ± 1, 8 ± 3, 4.5 ± fold increase compared to the corresponding control values for U251, Ln229, U87, XD456, JX6 cell lines, respectively, P<0.05, n=3.

Article Snippet: The PDL1 level in CSF samples was analyzed by using the Mouse PDL1 ELISA Kit (Boster, Pleasanton, CA, USA).

Techniques: Western Blot, Control

Journal: Journal of cancer science & therapy

Article Title: Blocking PD1/PDL1 Interactions Together with MLN4924 Therapy is a Potential Strategy for Glioma Treatment.

doi: 10.4172/1948-5956.1000543

Figure Lengend Snippet: Figure 4: Glioma cells treated with MLN4924 decrease T-cell proliferation via utilization of PD1/PDL1 signaling pathway. A) The graph illustrates an inhibition of T-cell proliferation after T-cell encounter with U251 cells (1); with U251 cells in the presence of an inhibitor of PD1/PDL1 interaction (2); with U251 cells treated with MLN4924 (1uM, for 4 days) (3); with U251 cells treated with MLN4924 (1uM, for 4 days) and in the presence of an inhibitor of PD1/PDL1 interaction (4). After MLN4924 treatment, glioma cells were washed and placed in the media with/and without an inhibitor of PD1/PDL1 interaction (4 uM). Note, that glioma cells treated with MLN4924 induce a stronger decrease of T-cell proliferation compared to untreated cells (55 ± 8% (n=4) versus 30 ± 8% (n=4), respectively, the difference is significant with P=0.0005). The reduction of T-cell proliferation induced by glioma cells treated with MLN4924 is inhibited in the presence of an inhibitor of PD1/PDL1 interaction (P=0.0009, n=4). B) The graph illustrates an inhibition of T-cell proliferation after T-cell encounter with XD456 cells (1); with UXD456 cells in the presence of an inhibitor of PD1/PDL1 interaction (2); with XD456 cells treated with MLN4924 (1uM, for 4 days) (3); with XD456 cells treated with MLN4924 (1uM, for 4 days) and in the presence of an inhibitor of PD1/PDL1 interaction (4). After MLN4924 treatment, glioma cells were washed and placed in media with/and without an inhibitor of PD1/PDL1 interaction. Note, that glioma cells treated with MLN4924 induce a stronger decrease of T-cell proliferation compared to untreated cells (74 ± 8% (n=4) versus 41 ± 7% (n=4), respectively, the difference is significant with P=0.0003). The reduction of T-cell proliferation induced by glioma cells treated with MLN4924 is inhibited in the presence of an inhibitor of PD1/PDL1 interaction (P=0.007, n=4).

Article Snippet: The PDL1 level in CSF samples was analyzed by using the Mouse PDL1 ELISA Kit (Boster, Pleasanton, CA, USA).

Techniques: Inhibition

Journal: The American Journal of Clinical Nutrition

Article Title: Effects of supplementation with nondigestible carbohydrates on fecal calprotectin and on epigenetic regulation of SFRP1 expression in the large-bowel mucosa of healthy individuals ^{1 2}

doi: 10.3945/ajcn.116.135657

Figure Lengend Snippet: Effects of supplementation with RS and PD on fecal calprotectin concentration ¹

Article Snippet: Fecal calprotectin was quantified by using the CALPROLAB Calprotectin ELISA (ALP) kit (Calpro AS) according to the manufacturer’s instructions by using fecal extracts diluted 1:20 in sample dilution buffer.

Techniques: Concentration Assay

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Schematic illustration of US-triggered deep-tissue activation of SPINs to release immunomodulators. b Schematic illustration of sonodynamic activation of SPINs to debulk tumor, enhance tumor immunogenicity, and release immunomodulators in situ as well as synergetic action of IDO inhibition and PD-L1 blocking on enhancing antitumor immunity with alleviated irAEs relative to free-drug administration.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Activation Assay, In Situ, Inhibition, Blocking Assay

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Chemical structures of amphiphilic semiconducting polymeric modulators and schematic illustration of their self-assembly and surface modification to form SPINs. b The molar ratios of each component in different SPINs. c Zeta potentials and hydrodynamic sizes of different SPINs in 1× PBS buffer (pH = 7.4) ( n = 4). d Photographs of erythrocytes after incubation with 1× PBS buffer (negative control), 1% Triton X-100 (positive control), and 1× PBS buffer containing SPINs at the concentration of 100 µg/mL for 2 h, followed by centrifugation. e Hemolysis percentages of erythrocytes after incubation with SPINs at different concentrations for 2 h ( n = 4). f Schematic illustration of US irradiation of SPIN D2 solutions covered with a pork tissue. g ESR spectra of 1 O 2 for SPIN D2 (20 µg/mL) after US irradiation (1.2 W/cm 2 , 3 min) without or with coverage of pork tissues at different thicknesses. h Release profiles of aPD-L1 and NLG919 from SPIN D2 (40 µg/mL) after US irradiation for different time ( n = 4). i PD-L1/PD-1 binding activity assay after treatment with free aPD-L1 or SPIN D2 (40 µg/mL) with or without US irradiation ( n = 4). SPIN D2 – US versus SPIN D2 + US: P < 0.0001. Statistical significance was calculated via a two-tailed Student’s t test. *** P < 0.001. In ( g – i ), the power intensity of US irradiation was 1.2 W/cm 2 (1.0 MHz, 50% duty cycle). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Modification, Incubation, Negative Control, Positive Control, Concentration Assay, Centrifugation, Irradiation, Binding Assay, Activity Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Schedule for the establishment of primary and distant tumors, triple systemic injection of SPINs (0.2 mL, 0.6 mg/mL) via tail vein, US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min), and analysis of immune responses. b , c Relative tumor volumes of primary ( b ) and distant ( c ) tumors of Panc02 tumor-bearing C57BL/6 mice ( n = 6) after systemic injection of saline, free-drug mixture (on day 0, 3, and 6, 4 mg/kg body weight for NLG919 and aPD-L1), or SPIN D2 (0.2 mL, 0.6 mg/mL) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min). SPIN D2 + US versus drug + US: P < 0.0001 for primary tumors ( b ); SPIN D2 + US versus drug: P < 0.0001 for distant tumors ( c ). d Survival curves of Panc02 tumor-bearing C57BL/6 mice ( n = 10) receiving different treatments as indicated. e Schematic illustration of treatment of rechallenged tumor mouse models using SPINs. f Growths of rechallenged tumors in Panc02 tumor-bearing mice after injection of saline or SPIN D2 (0.2 mL, 0.6 mg/mL) with US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min) ( n = 5). Saline versus SPIN D2 + US: P < 0.0001. g The survival curves of Panc02 tumor-bearing C57BL/6 mice after different treatments followed by tumor rechallenge ( n = 10). h Flow cytometry analysis of populations of effector memory T cells in the spleen of Panc02 tumor-bearing C57BL/6 mice after different treatments followed by tumor rechallenge ( n = 4). Saline versus SPIN D2 + US: P = 0.0059. i Differentially expressed gene numbers in tumor tissues of mice after different treatments. j Relative expression of Carl , Hmgb1-ps1 , Hmgb1-ps2 , Cd80 , Cd86 , Cd40 , Pdcd1 , Cd3e , Cd8a , Ifng , Gzmb , Cxcl1 , Cxcl2 , Cxcl9 , Cxcl10 , Cxcl11 , Ccl4 , Ccl5 , Il1b , Il2 , Il6 , Il7 , Il15 , Ido1 , and Cd274 in tumors of Panc02 tumor-bearing mice after different treatments (the experiment was repeated independently five times with similar results). k Unsupervised hierarchical clustering of relative gene expression in tumors of Panc02 tumor-bearing C57BL/6 mice after different treatments ( n = 5). Data are presented as mean values ± SD. Statistical significance was calculated via two-tailed Student’s t test; ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Injection, Irradiation, Flow Cytometry, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Schematic of sono-immunotherapy of subcutaneous pancreatic mouse tumors covered with 5-cm tissue. b , c Relative tumor volumes of primary ( b ) and distant ( c ) tumors of Panc02 tumor-bearing C57BL/6 mice ( n = 5) after systemic injection of saline, free-drug mixture (on day 0, 3, and 6, 4 mg/kg body weight for NLG919 and aPD-L1), SPIN 0 or SPIN D2 (0.2 mL, 0.6 mg/mL) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min). The primary tumors were covered with 5-cm tissue under US irradiation. SPIN D2 + US versus SPIN 0 + US: P < 0.0001 for primary tumors ( b ); SPIN D2 + US versus SPIN 0 + US: P < 0.0001 for distant tumors ( c ). d Survival curves of Panc02 tumor-bearing C57BL/6 mice ( n = 10) after different treatments for 60 days. e Schematic of US-mediated deep-tissue sonodynamic therapy of orthotopic pancreatic rabbit tumors. f Radiolabeling stability of 131 I-SPIN 0 after storage in saline or 50% serum at 37 °C for different time ( n = 3). g , h SPECT imaging ( g ) and signal intensity ( h ) of orthotopic pancreatic rabbit tumors after systemic injection of 131 I-SPIN 0 (1.0 mL, 1.5 mg/mL) for different time ( n = 4). The white dotted circle indicated tumors. i Computed tomography (CT) imaging of orthotopic pancreatic rabbit tumors after systemic injection of saline or SPIN 0 (1.0 mL, 1.5 mg/mL) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 30 min). The white dotted circle indicated tumors. j Tumor volume of orthotopic pancreatic rabbit tumors ( n = 3) after treatments as indicated for different days. Saline + US versus SPIN 0 + US: P = 0.0108. k H&E staining images of orthotopic pancreatic rabbit tumors after different treatments. The experiment was repeated independently three times with similar results. l Survival curves of orthotopic pancreatic tumor-bearing rabbits ( n = 4) after different treatments for 20 days. Data are presented as mean values ± SD. Statistical significance was calculated via two-tailed Student’s t test; * P < 0.05, *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Injection, Irradiation, Radioactivity, Single Photon Emission Computed Tomography, Imaging, Computed Tomography, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a , b Flow cytometry analysis of percentages of CD3 + CD4 + Th cells ( a ), and CD3 + CD8 + CTLs ( b ) in blood of mice ( n = 4) at 30 day after systemic administrations of saline, SPIN 0 , SPIN D2 (0.2 mL, 1.2 mg/mL) or free-drug mixture (8 mg/kg body weight for NLG919 and aPD-L1) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min). Saline – US versus drug − US: P = 0.0023; saline − US versus drug + US: P = 0.0006; drug + US versus SPIN D2 + US: P = 0.0071 for CD3 + CD4 + Th cells ( a ); saline − US versus drug − US: P = 0.0004; saline − US versus drug + US: P = 0.0001; drug + US versus SPIN D2 + US: P = 0.0093 for CD3 + CD8 + CTLs ( b ). c , d Flow cytometry analysis of percentages of CD3 + CD4 + Th cells ( c ), and CD3 + CD8 + CTLs ( d ) in spleen of mice ( n = 4) after different treatments for 30 days. Saline − US versus drug − US: P = 0.0008; saline − US versus drug + US: P = 0.0005; drug + US versus SPIN D2 + US: P = 0.0015 for CD3 + CD4 + Th cells ( c ); saline − US versus drug − US: P = 0.0001; saline − US versus drug + US: P = 0.0002; drug + US versus SPIN D2 + US: P = 0.0049 for CD3 + CD8 + CTLs ( d ). e Representative H&E staining images of liver after 30 days of treatments in different groups (white arrows indicate the infiltrated lymphocytes). The experiments were repeated independently three times with similar results. f Heatmap to show relative fold of cytokine levels in serum of mice after different treatments for 30 days relative to those in saline control group. g , h Serum levels of ALT ( g ) and AST ( h ) in mice ( n = 5) after different treatments for 30 days. Saline − US versus drug − US: P = 0.0010; saline − US versus drug + US: P = 0.0020; drug + US versus SPIN D2 + US: P = 0.0054 for ALT ( g ); saline − US versus drug − US: P = 0.0001; saline − US versus drug + US: P < 0.0001; drug + US versus SPIN D2 + US: P = 0.0013 for AST ( h ). i Summary comparison of the antitumor immunity and irAEs between SPIN D2 -mediated sono-immunotherapy and free-drug treatment. Data are presented as mean values ± SD. Statistical significance was calculated via two-tailed Student’s t test; ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Flow Cytometry, Irradiation, Staining, Two Tailed Test

Journal: Journal of Cancer

Article Title: Inositol hexaphosphate sensitizes hepatocellular carcinoma to oxaliplatin relating inhibition of CCN2-LRP6-β-catenin-ABCG1 signaling pathway

doi: 10.7150/jca.62141

Figure Lengend Snippet: IP6 directly inhibited the ability of proliferation and migration in HCC. (A) Cell viability assays of MHCC97L and Hep3B cells treated with oxaliplatin and/or IP6 exhibited synergistically enhanced anti-tumor efficacy. (B) Adherent colony formation assays of MHCC97L and Hep3B cells treated with IP6 exhibited inhibited proliferation ability. (C) IP6 significantly inhibited sphere formation in MHCC97L and Hep3B cells. (D) The transwell assay demonstrated that MHCC97L and Hep3B cells migrated through the basement membrane less efficiently when treated with IP6. (E) IP6 attenuate ability of HCC proliferation with reduced subcutaneous tumor weights in IP6 group when compared to control group. (F) IP6 attenuate HCC pulmonary metastatic potential with reduced number of pulmonary nodules in IP6 group when compared to control group.

Article Snippet: HCC cells were seeded into 96-well plates and treated with oxaliplatin at increasing concentrations (0, 0.5, 1, 2, 4, 8, 16, 32, 64, 128 μM) for 24, 48, 72, and 96 h. Cell proliferation assays were carried out with the Cell Counting Kit 8 (CCK8; Dojindo Molecular Technologies, Inc.).

Techniques: Migration, Transwell Assay, Membrane, Control

Journal: Journal of Cancer

Article Title: Inositol hexaphosphate sensitizes hepatocellular carcinoma to oxaliplatin relating inhibition of CCN2-LRP6-β-catenin-ABCG1 signaling pathway

doi: 10.7150/jca.62141

Figure Lengend Snippet: IP6 exhibited a synergistic and sequential anti-proliferative effect with oxaliplatin. (A) The response of HCC cells to combined therapy with different concentrations of oxaliplatin as assessed via measurement of half maximal inhibitory concentration (IC50) values, and IP6 also exhibited synergistically enhanced anti-tumor efficacy with lower IC50 values. (B) IP6-pretreated HCC cells exhibited increased intercellular adhesion and epithelioid cell morphology, which is contrary to the cell morphology from oxaliplatin-pretreated HCC cells. (C) IP6 pretreated HCC cells exhibited decreased oxaliplatin IC 50 values. (D) The oxaliplatin-pretreated HCC cell lines were then treated with IP6, yielding significantly reduced resistance to oxaliplatin with decreased IC 50 values in both MHCC97L-OXA and Hep3B-OXA.

Article Snippet: HCC cells were seeded into 96-well plates and treated with oxaliplatin at increasing concentrations (0, 0.5, 1, 2, 4, 8, 16, 32, 64, 128 μM) for 24, 48, 72, and 96 h. Cell proliferation assays were carried out with the Cell Counting Kit 8 (CCK8; Dojindo Molecular Technologies, Inc.).

Techniques: Concentration Assay

Journal: Journal of Cancer

Article Title: Inositol hexaphosphate sensitizes hepatocellular carcinoma to oxaliplatin relating inhibition of CCN2-LRP6-β-catenin-ABCG1 signaling pathway

doi: 10.7150/jca.62141

Figure Lengend Snippet: ABCG1 and CCN2 expression was upregulated by oxaliplatin and significantly reversed by IP6. (A) Immunoblot analysis verified that the expression of ABCG1 and CCN2 was significantly upregulated by oxaliplatin in a time- and concentration-dependent manner. (B) HCC cells treated with IP6 exhibited downregulated ABCG1 and CCN2 expression levels in a time- and concentration-dependent manner. (C) Tumor tissues from subcutaneous xenografts by immunohistochemistry exhibited a significantly decreased proportion of ABCG1 and CCN2 in IP6 treatment group.

Article Snippet: HCC cells were seeded into 96-well plates and treated with oxaliplatin at increasing concentrations (0, 0.5, 1, 2, 4, 8, 16, 32, 64, 128 μM) for 24, 48, 72, and 96 h. Cell proliferation assays were carried out with the Cell Counting Kit 8 (CCK8; Dojindo Molecular Technologies, Inc.).

Techniques: Expressing, Western Blot, Concentration Assay, Immunohistochemistry

Journal: Journal of Cancer

Article Title: Inositol hexaphosphate sensitizes hepatocellular carcinoma to oxaliplatin relating inhibition of CCN2-LRP6-β-catenin-ABCG1 signaling pathway

doi: 10.7150/jca.62141

Figure Lengend Snippet: Inhibition of ABCG1 reversed oxaliplatin resistance in HCC cells. (A) Oxaliplatin IC50 values decreased both in MHCC97L-ABCG1-sh1 and MHCC97L-OXA-ABCG1-Sh1 compared to the corresponding mock cells, and the same trend was confirmed in Hep3B-ABCG1-sh1 and Hep3B-OXA-ABCG1-Sh1. (B) Oxaliplatin IC 50 values still decreased in MHCC97L-OXA-ABCG1-Sh1 synergistically treated with IP6 compared to the control group.

Article Snippet: HCC cells were seeded into 96-well plates and treated with oxaliplatin at increasing concentrations (0, 0.5, 1, 2, 4, 8, 16, 32, 64, 128 μM) for 24, 48, 72, and 96 h. Cell proliferation assays were carried out with the Cell Counting Kit 8 (CCK8; Dojindo Molecular Technologies, Inc.).

Techniques: Inhibition, Control