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Fig 1.

Representative photographs of classification of mode of invasion (H&E staining, immunohistochemical pancytokeratin staining, x40 magnification).

(1A,1B) grade 1, (1C,1D) grade 2, (1E,1F) grade 3, (1G,1H) grade 4C, and (1I,1J) grade 4D.

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Fig 2.

Representative photographs of classification of worst pattern of invasion (H&E staining, immunohistochemical pancytokeratin staining, x200 magnification).

(2A,2B) Worst pattern of invasion (WPOI)-1, (2C,2D) WPOI-2, (2E,2F) WPOI-3, (2G,2H) WPOI-4, and (2I,2J, x40 magnification) WPOI-5.

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Fig 3.

Representative photographs of intensity of tumor budding (immunohistochemical pancytokeratin staining, x200 magnification).

(3A) Low intensity of tumor budding (<5 buds), (3B) intermediate intensity of tumor budding (≥5~<10 buds), and (3C) high intensity of tumor budding (≥10 buds).

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Fig 3 Expand

Fig 4.

Photographs of immunohistochemical pancytokeratin staining show representative examples of grade 4D mode of invasion and worst pattern of invasion (WPOI)-4 at x40 magnification.

The region in the rectangle is shown at x200 magnification in the lower panel, which shows 12 buds.

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Fig 4 Expand

Fig 5.

Photographs of immunohistochemical pancytokeratin staining show representative examples of grade 3 mode of invasion and WPOI-5 at x12.5 magnification.

The yellow line measures a distance >1 mm between the main tumor and the next focus of dispersed islands. The region in the rectangle is shown at x200 magnification in the lower panel, which shows 1 bud.

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Fig 6.

Kaplan-Meier survival curves by mode of invasion (A), worst pattern of invasion (WPOI) (B), tumor budding (C), lymphovascular invasion (D), and perineural invasion (E).

5-year disease-free survivals were 58.0% for grade 1+2+3 mode of invasion, 36.4% for grade 4C mode of invasion, 0.0% for grade 4D mode of invasion (A), 58.4% for WPOI-1+2+3+4, 33.3% for WPOI-5, 65.0% for low-intensity tumor budding (<5 buds), 37.5% for intermediate-intensity tumor budding (≥5 buds-<10 buds), 23.1% for high-intensity tumor budding (≥10 buds), 58.1% for absence of lymphovascular invasion, 33.3% for presence of lymphovascular invasion, 56.1% for absence of perineural invasion, and 20.0% for presence of perineural invsion in patients with clinically node-negative T1 and T2 oral squamous cell carcinoma.

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Fig 7.

Kaplan-Meier curves of disease-free survival (DFS) in patients in combined groups with the intensity of tumor budding and mode of invasion (A) or worst pattern of invasion (WPOI) (B).

In group A, the 5-year DFSs were 70.4% with low tumor budding and grade 1+2+3 mode of invasion ①, 66.7% with low tumor budding and grade 4C mode of invasion ②, 50.0% with intermediate tumor budding and grade 1+2+3 mode of invasion ③, 0% with intermediate tumor budding and grade 4C mode of invasion ④, 20.0% with high tumor budding and grade 1+2+3 mode of invasion ⑤, 33.3% with high tumor budding and grade 4C mode of invasion ⑥, and 0% with high tumor budding and grade 4D mode of invasion ⑦. In group B, the 5-year DFSs were 72.8% with low tumor budding and WPOI-1+2+3+4 ①, 44.4% with low tumor budding and WPOI-5 ②, 36.4% with intermediate tumor budding and WPOI-1+2+3+4 ③, 60.0% with intermediate tumor budding and WPOI-5 ④, 33.3% with high tumor budding and WPOI-1+2+3+4 ⑤, and 0% for high tumor budding and WPOI-5 ⑥.

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