Objective. To analyze the active compounds, potential targets, and diseases of JianPi Fu Recipe (JPFR) based on network pharmacology and bioinformatics and verify the potential biological function and mechanism of JPFR in vitro and in vivo. Methods. Network pharmacology databases including TCMSP, TCM-PTD, TCMID, and DrugBank were used to screen the active compounds and potential drug targets of JPFR. Cytoscape 3.7 software was applied to construct the interaction network between active compounds and potential targets. The DAVID online database analysis was performed to investigate the potential effective diseases and involved signaling pathways according to the results of the GO function and KEGG pathways enrichment analysis. To ensure standardization and maintain interbatch reliability of JPFR, High Performance Liquid Chromatography (HPLC) was used to establish a “chemical fingerprint.” For biological function validation, the effect of JPFR on the proliferation and migration of CRC cells in vitro was investigated by CCK-8 and transwell and wound healing assay, and the effect of JPFR on the growth and metastasis of CRC cells in vivo was detected by building a lung metastasis model in nude mice and in vivo imaging. For the potential mechanism validation, the expressions of MALAT1, PTBP-2, and β-catenin in CRC cells and transplanted CRC tumors were detected by real-time PCR, western blot, and immunohistochemical staining analysis. Results. According to the rules of oral bioavailability (OB) > 30% and drug-likeness (DL) > 0.18, 244 effective compounds in JPFR were screened out, as well as the corresponding 132 potential drug targets. By the analysis of DAVID database, all these key targets were associated closely with the cancer diseases such as prostate cancer, colorectal cancer, bladder cancer, small cell lung cancer, pancreatic cancer, and hepatocellular carcinoma. In addition, multiple signaling pathways were closely related to JPFR, including p53, Wnt, PI3K-Akt, IL-17, HIF-1, p38-MAPK, NF-κB, PD-L1 expression and PD-1 checkpoint pathway, VEGF, JAK-STAT, and Hippo. The systematical analysis showed that various active compounds of JPFR were closely connected with Wnt/β-catenin, EGFR, HIF-1, TGFβ/Smads, and IL6-STAT3 signaling pathway, including kaempferol, isorhamnetin, calycosin, quercetin, medicarpin, phaseol, spinasterol, hederagenin, beta-sitosterol, wighteone, luteolin, and isotrifoliol. For in vitro experiments, the migration and growth of human CRC cells were inhibited by the JPFR extract in a dose-dependent way, and the expression of MALAT1, PTBP-2, β-catenin, MMP7, c-Myc, and Cyclin D1 in CRC cells were downregulated by the JPFR extract in a dose-dependent way. For in vivo metastasis experiments, the numbers of lung metastasis were found to be decreased by the JPFR extract in a dose-dependent manner, and the expressions of metastasis-associated genes including MALAT1, PTBP-2, β-catenin, and MMP7 in the lung metastases were downregulated dose dependently by the JPFR extract. For the orthotopic transplanted tumor experiments, the JPFR extract could inhibit the growth of orthotopic transplanted tumors and downregulate the expression of c-Myc and Cyclin D1 in a dose-dependent manner. Moreover, the JPFR extract could prolong the survival time of tumor-bearing mice in a dose-dependent manner. Conclusions. Through effective network pharmacology analysis, we found that JPFR contains many effective compounds which may directly target cancer-associated signaling pathways. The in vitro and in vivo experiments further confirmed that JPFR could inhibit the growth and metastasis of CRC cells by regulating β-catenin signaling-associated genes or proteins.

The objective of this study was to evaluate the effect of exogenous ghrelin on the behaviors, weight gain, and feed intakes of weanling piglets. A total of 25 pairs of Duroc×Landrace×Large White piglets weaned at 21 days of age were used in this experiment which finished on day 36. Each pair of healthy piglets from the same litter with similar body weight and of the same gender were selected and randomly arranged to ghrelin or control groups. Thus, there were 50 piglets (ghrelin 25; control 25) kept in 10 pens (ghrelin 5; control 5) and 5 piglets per pen. Initial body weight of the pigs did not differ between the control and ghrelin treatment (7.43±0.17 kg; p = 0.81). Experimental pigs were infused with ghrelin (1 μg/d pig) via the marginal ear vein between 0750 and 0800 h at 22, 23, 24 days of age. Control pigs were infused with 0.9% saline. Feed consumption was measured on days 23, 24, 25, 29 and 36. Body weight was measured on days 22, 23, 24, 25, 29 and 36. Behavior data of individual piglets were collected by real-time observation from 0800-1500 h through remote supervisory equipment at 22, 23 and 24 days of age. The results indicated that ghrelin infusion could increase drinking (p<0.05) and lying behaviors (p<0.01) and decrease mounting behaviors (p<0.05). No significant influence of ghrelin was found on average daily weight gain (ADG) and average daily feed intake (ADFI) in this experiment (p>0.05). In conclusion, exogenous ghrelin by the method above and at the dosage of 1 μg/d pig could cause a variety of behavioral effects, but not improve performance of weanling piglets.

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ObjectiveBufalin, the main active anti-tumor monomer of toad venom, is crucial in cancer treatment. However, intrinsic issues, such as poor solubility and systematic toxicity, have considerably mitigated its anticancer functions and caused unwanted side effects. It is essential to develop innovative targeting systems to precisely and efficiently deliver anticancer drugs to achieve satisfying therapeutic efficiency.MethodsThis work established a novel and more efficient system for simultaneously detecting and killing colorectal cancer cells. The proposed method designed two allosteric probes, a report probe and a recognize probe. The method exhibited high sensitivity towards cell detection via the recognizing probe identifying target cancer cells and the report probe's signal report. Combining bufalin and fluorouracil endowed better tumor cell inhibition.ResultsWe observed significantly enhanced fluorescence dots surrounding the HCT-116 cell membranes. No fluorescence increments in the other three cells were identified, indicating that the established liposome complex could specifically bind with target cells. In addition, the best ratio of bufalin to fluorouracil was 0.15 and 0.5, respectively. This improved the anti-tumor effects and achieved more than 60% tumor cell inhibition.ConclusionThis method will provide new opportunities for intracellular biomolecule detection and targeted cancer cell therapy.