1. Volume 128, Issue 7, Pages 1965-1983 (June 2005)
Synergistic Inhibitory Effects of Gastrin and Histamine Receptor Antagonists on Helicobacter-Induced Gastric Cancer 
Shigeo Takaishi, Guanglin Cui, Dana M. Frederick, Jane E. Carlson, JeanMarie Houghton, Andrea Varro, Graham J. Dockray, Zhongming Ge, Mark T. Whary, Arlin B. Rogers, James G. Fox, Timothy C. Wang 
Gastroenterology 
Volume 128, Issue 7, Pages (June 2005)
DOI: /j.gastro
Copyright © 2005 American Gastroenterological Association Terms and Conditions

2. Figure 1
YF476 and/or loxtidine treatment for 3 months strongly inhibited gastric acid output in H felis-infected INS-GAS mice. Gastric acid output measured by the pyloric ligation method; acid secretion over 4 hours was determined by the pyloric ligation method and expressed as microequivalent protons. H felis-infected INS-GAS mice treated with YF476 and/or loxtidine for 3 months showed a significant decrease of gastric acid output compared with mice receiving no drug. (*P < .05; n = 4 per each group). LOX, loxtidine; YF + LOX, YF476 plus loxtidine; FVB, untreated FVB/N control mice.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

3. Figure 2
YF476 and/or loxtidine treatment for 3 months synergistically inhibited gastric atrophy, hyperplasia, and dysplasia in H felis-infected INS-GAS mice. Representative H&E stains are shown (original magnification 60×; scale bar = 250 μm). (A) No drug, (B) YF476, (C) loxtidine, and (D) YF476 plus loxtidine. Treatment with (B) YF476 alone or (C) loxtidine alone for 3 months resulted in partial inhibition of gastric atrophy and foveolar hyperplasia noted in H felis-infected INS-GAS mice receiving (A) no drug. In addition, the mice treated with (D) YF476 plus loxtidine for 3 months showed almost complete inhibition of gastric atrophy, hyperplasia, and dysplasia.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

4. Figure 3
Serum amidated gastrin levels of H felis-infected INS-GAS mice treated with YF476 and/or loxtidine for 6 months. Serum amidated gastrin levels of H felis-infected INS-GAS mice treated with YF476 alone or YF476 plus loxtidine for 6 months were significantly higher than mice receiving no drug (*P < .05; **P < .01), whereas those of mice treated with loxtidine alone were not significant, but the values tended to be higher when compared with serum amidated gastrin levels of mice receiving no drug (#P = .054). LOX, loxtidine; YF + LOX, YF476 plus loxtidine (n = 10 for each group).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

5. Figure 4
YF476 and/or loxtidine treatment for 6 months synergistically inhibited gastric tumors in H felis-infected INS-GAS mice. (A–E) Gross presentation of the stomachs of H felis-infected INS-GAS mice treated with YF476 and/or loxtidine for 6 months. (A) No drug, (B) YF476, (C) loxtidine, (D) YF476 plus loxtidine, and (E) untreated FVB/N control mice. The stomachs of H felis-infected INS-GAS mice treated with (B) YF476 alone or (C) loxtidine alone for 6 months were smaller than those of mice receiving (A) no drug, and the size of the stomachs of mice treated with (D) YF476 plus loxtidine was almost same as those of (E) untreated FVB/N control mice. Each distance between 2 adjoining bars in the scale represents 1 mm. (F–H) Stomach wet weight, body weight, and the ratio of stomach wet weight over body weight of H felis-infected INS-GAS mice treated with YF476 and/or loxtidine for 6 months. (F) Stomach wet weight and (H) the ratio of stomach wet weight over body weight of mice treated with YF476 and/or loxtidine for 6 months were significantly smaller than those of mice receiving no drug (*P < .01; n = 6 per each group), whereas (G) body weight of those mice was significantly larger than that of mice receiving no drug. (*P < .01; n = 6 per each group). LOX, loxtidine; YF + LOX, YF476 plus loxtidine; FVB, untreated FVB/N control mice.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

6. Figure 5
YF476 and/or loxtidine treatment for 6 months synergistically inhibited gastric carcinogenesis in H felis-infected INS-GAS mice. Representative H&E stains are shown (original magnification 60×; scale bar = 250 μm). (A) No drug, (B) YF476, (C) loxtidine, and (D) YF476 plus loxtidine. Treatment with (B) YF476 alone or (C) loxtidine alone for 6 months resulted in a significant decrease in overall mucosal thickness, a partial inhibition of neoplasia, and the elimination of submucosal invasion observed in H felis-infected INS-GAS mice receiving (A) no drug. In addition, treatment with (D) YF476 plus loxtidine for 6 months resulted in nearly complete inhibition of neoplasia and normalization of histology, with only mild inflammation and edema as noted in mice treated with (B) YF476 alone or (C) loxtidine alone.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

7. Figure 6
Immunohistochemical studies of ECL cells and parietal cells in H felis-infected INS-GAS mice treated with YF476 and/or loxtidine for 6 months. (A and B) Immunohistochemical staining of ECL cells: (A) no drug and (B) YF476 plus loxtidine treatment for 6 months (anti-chromogranin A antibody; original magnification 100×). (C and D) Immunohistochemical staining of parietal cells: (C) no drug and (D) YF476 plus loxtidine treatment for 6 months (anti—H+,K+-adenosine triphosphatase β subunit antibody; original magnification 100×).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

8. Figure 7
H felis infection status in the stomachs of YF476- and/or loxtidine-treated H felis-infected INS-GAS mice. (A) Warthin-Starry silver staining of H felis in the antrum of stomach of H felis-infected INS-GAS mice treated with YF476 plus loxtidine for 6 months; H felis were detected as spiral-formed bacteria (original magnification 1000×). (B) ELISA assay for mice measuring serum H felis-specific IgG antibody. H felis IgG titers indicated minor changes among mice treated with YF476 and/or loxtidine for 6 months and mice receiving no drug (n = 6 per each group). (C) Quantitative real-time PCR analysis for H felis DNA from the gastric corpus DNA of H felis-infected INS-GAS mice. Increased H felis DNA copy numbers per gastric corpus DNA (copy per microgram) were not significant but were slightly increased in mice treated with YF476 and/or loxtidine for 6 months compared with mice receiving no drug (n = 6 per each group). LOX, loxtidine; YF + LOX, YF476 plus loxtidine; FVB, uninfected FVB/N control mice.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

9. Figure 8
Growth factor expression analysis in YF476- and/or loxtidine-treated H felis-infected INS-GAS mice. Quantitative real-time RT-PCR analysis of growth factor expression level: (A) Reg I, (B) amphiregulin, (C) HB-EGF, and (D) TGF-α. All 3 groups of mice treated with YF476 and/or loxtidine for 6 months showed a significantly lower expression level of (A) Reg I and (B) amphiregulin than mice receiving no drug, whereas the expression level of (C) HB-EGF was not significant but showed a tendency for lower values. (D) TGF-α expression did not show a significant change in response to YF476 and/or loxtidine treatment (*P < .05, **P < .01, n = 6 for each group). LOX, loxtidine; YF + LOX, YF476 plus loxtidine; INSGAS, INS-GAS mice without H felis infection; FVB, untreated FVB/N control mice.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

10. Figure 9
Loxtidine treatment of H felis-infected INS-GAS mice showed mild shifts of cytokine expression profiles from Th1 to Th2 polarization. (A–D) Quantitative real-time RT-PCR analysis of Th1, Th2 cytokines and somatostatin expression level: (A) IFN-γ, (B) TNF-α, (C) IL-4, and (D) somatostatin (*P < .05; **P < .01; N.D., not detected; n = 6 for each group). Mice treated with loxtidine alone or YF476 plus loxtidine for 6 months showed a significantly lower expression level of (A) IFN-γ and (B) TNF-α compared with mice receiving no drug (*P < .05), whereas mice treated with YF476 alone for 6 months did not show a significant change. Similarly, mice treated with loxtidine alone or YF476 plus loxtidine for 6 months showed significantly higher expression levels of (C) IL-4 and (D) somatostatin compared with mice receiving no drug (*P < .05), whereas mice treated with YF476 alone for 6 months did not show a significant change. (E) Serum H felis IgG1/IgG2a ratio (n = 6 for each group). Serum H felis IgG1/IgG2a ratio of mice treated with YF476 alone or loxtidine alone for 6 months was significantly lower than of mice receiving no drug (*P < .05, **P < .01), whereas mice treated with YF476 plus loxtidine for 6 months did not show a significant change, although there was a tendency for higher values (#P = .054). LOX, loxtidine; YF + LOX, YF476 plus loxtidine; INSGAS, INS-GAS mice without H felis infection; FVB, untreated FVB/N control mice.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

11. Figure 10
Omeprazole treatment for 3 months resulted in the mild progression of gastric hyperplasia and dysplasia in H felis-infected INS-GAS mice. Representative H&E stains are shown (original magnification 60×; scale bar = 250 μm). (A) No drug, (B) omeprazole alone, (C) omeprazole plus YF476, and (D) omeprazole plus loxtidine. Treatment with (B) omeprazole alone for 3 months did not show a reduction in atrophy but instead appeared to manifest a more rapid progression of gastric foveolar hyperplasia and dysplasia than H felis-infected INS-GAS mice receiving (A) no drug, whereas the combination of omeprazole with (C) YF476 or (D) loxtidine for 3 months resulted in a significant suppression of gastric hyperplasia and dysplasia compared with mice treated with omeprazole alone.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions

12. Figure 11
Serum gastrin levels and immunohistochemical studies of ECL cells and parietal cells in H felis-infected INS-GAS mice treated with omeprazole alone or in combination with YF476 or loxtidine for 3 months. (A) Serum amidated gastrin levels of H felis-infected INS-GAS mice treated with omeprazole alone or in combination with YF476 and/or loxtidine for 3 months. Serum amidated gastrin levels of H felis-infected INS-GAS mice treated with omeprazole alone or in combination with YF476 or loxtidine for 3 months were significantly higher than mice receiving no drug (*P < .05). OMP, omeprazole alone; OMP + YF, omeprazole plus YF476; OMP + LOX, omeprazole plus loxtidine; n = 5 for each group. (B and C) Immunohistochemical staining of ECL cells: (B) no drug and (C) treated with omeprazole alone for 3 months (anti-chromogranin A antibody; original magnification 100×). (D and E) Immunohistochemical staining of parietal cells: (D) no drug and (E) treated with omeprazole alone for 3 months (anti-H+,K+-adenosine triphosphatase β subunit antibody; original magnification 100×).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2005 American Gastroenterological Association Terms and Conditions