Trypsin

Candida utilis was purchased from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (No. DSM 2361). Total superoxide dismutase (SOD), total antioxidant capacity, aminotransferase aspartate, alanine aminotransferase, glutathione peroxidase, glucose, total cholesterol, total protein, trypsin, amylase, and lipase were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Rabbit-actin, occludin, ZO-1, β-defensin-2, and goat-rabbit all purchased from Cell Signaling Technology (Danvers, America). Seal fluid and developer were purchased from Millipore. AMV First Strand cDNA Synthesis Kit was purchased from Shanghai Bioengineering (Shanghai, China). SYBR^® Green PCR Kit and PowerFecal DNA Isolation Kit was purchased from Germany Qiagen reagent company (Dusseldorf, Germany).

The intestines of five fish fries were pooled to make one sample, and a total four samples were collected at each stage to determine digestive enzymatic activity. All the samples were homogenized in 2 mL of chilled PBS buffer on ice, and then centrifuged at 3,000 × g for 10 min at 4°C to obtain a clear supernatant (3–18 K, Sigma^®, Germany). Finally, the clear supernatant was used for quantitative assays of trypsin, lipase, and amylase activities, and total protein. The enzyme activities and total protein were measured by detection kits purchased from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China), which included trypsin, lipase, amylase, and total protein assay kits (Product codes: A080-2-2, A054-1-1, C016-1-1, and A045-3-2, respectively).

Paclitaxel was ordered from Xi’an Natural Field Bio-Technique Co, Ltd. (Xi’an, China) with >99% purity. Folic acid, N-hydroxysuccinimide (NHS), dicyclohexyl carbodiimide (DCC), dimethyl sulfoxide (DMSO), sodium dodecyl sulfate (SDS), ethanol, methanol, paraformaldehyde, and Pluronic 127 (F127) were purchased from Aladdin Chemical Reagent Co. (Shanghai, China). Thiazolyl blue tetrazolium bromide (MTT), propidium iodide (PI), Annexin V-FITC apoptosis detection kit, trypsin, Triton X-100, and bovine serum albumin were supplied by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). SMMC-7721 cell lines were obtained from the National Experimental Cell Resource Platform (Beijing, China). RPMI 1640 medium and fetal bovine serum (FBS) were purchased from Beijing Dingguo Changsheng Biotech Co. Ltd. (Beijing, China). Phenylmethanesulfonyl fluoride (PMSF), 4′,6-diamidino-2-phenylindole (DAPI), mitochondrial membrane potential assay kit (JC-1), calcein-AM/PI were provided by Beyotime Biotechnology Co. (Shanghai, China). Bcl-2, Bax, and cleaved caspase-3 were obtained from Abcam (Cambridge, UK). N-Cadherin, galectin-3, EpCAM, CD147, and EGFR were purchased from Wanleibio (Shenyang, China).

Doxorubicin (DOX, Mw = 543.52, purity ≥ 98%) was provided by Hefei Bomei Biological
Technology Co., Ltd. (Hefei, China). [2-(Acryloyoxyethyl)ethyl]trimethylammonium chloride
(DAC, 80 wt%) was purchased from Jinan Wanduoxin Chemical Co., Ltd. (Jinan, China). Cetyl
trimethyl ammonium bromide (CTAB) was obtained from Tianjin Guangfu Fine Chemical Co.,
Ltd. (Tianjin, China). 2,2′-Azodiazine(2-methylpropiamide)dihydrochloride (AIBA) was
obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Thiazolyl blue
tetrazolium bromide (MTT), Annexin V-FITC Apoptosis Detection Kit, trypsin, Triton x-100,
and bovine serum albumin were supplied by Nanjing Jiancheng Bioengineering Institute
(Nanjing, China). Folate, N-hydroxysuccinimide (NHS), dicyclohexyl carbodiimide (DCC),
dimethylsulfoxide (DMSO), ammonia, ethanol, methanol, and paraformaldehyde were purchased
from Sinopharm Holding Chemical Reagent Company, Ltd. (Shanghai, China). SMMC-7721 and H22
cell lines were obtained from the national experimental cell resource platform (Beijing,
China). Polyethylenimine (PEI, Mw = 25,000) was obtained from Aladdin Chemical Inc.
(Shanghai, China). Other reagents were of analytical/chromatographic grade, and distilled
water was used in all experiments.

Antibodies for Western blotting were purchased from Cell Signaling Technology (Shanghai, China). Gastrin (Gastrin), malondialdehyde (MDA), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), catalase (CAT), nitric oxide (NO), prostaglandin E2, myeloperoxidase (MPO), α-amylase, α-glucosidase, trypsin, a chymotrypsin kit, and an ELISA kit for interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).