Aim To investigate the inhibitory effect of the protein kinase D(PKD)-specific inhibitor CRT0066101 on hepatocellular carcinoma(HCC)and its underlying molecular mechanisms,providing new theoretical insights and therapeutic strategies for targe-ted HCC treatment.Methods HCC cell lines were treated with varying concentrations of CRT0066101.The inhibitory effects on cell proliferation were assessed using the CCK-8 assay,colony formation assay,and EdU staining.The impact on cell migration and inva-sion was evaluated through wound-healing assays and Transwell migration and invasion assays.was employed toThe effects of CRT0066101 on the phosphorylation levels of PKD and key proteins in the downstream PI3K/AKT signaling pathway were analyzed using Western blot.Additionally,the drug's regulatory effects on apoptosis and autophagy in HCC cells were examined using Western blot,flow cytometry,and the mRFP-GFP-LC3 dual-fluorescence reporter system.Results CRT0066101 significantly inhibited the pro-liferation,migration and invasion of HCC cells.West-ern blotting results demonstrated that CRT0066101 dose-dependently suppressed the phosphorylation of PKD family proteins and downregulated the activation of the PI3K/AKT signaling pathway.Furthermore,CRT0066101 treatment upregulated the expression of the pro-apoptotic protein Bax while downregulating the anti-apoptotic protein Bcl-2.It also markedly increased the expression levels of autophagy marker proteins Bec-lin-1 and LC3B-Ⅱ,suggesting that the drug simulta-neously induced apoptosis and autophagy in HCC cells.Conclusions CRT0066101 specifically inhibits PKD activity,blocks the PI3K/AKT signaling path-way,suppresses HCC cell proliferation and metastasis,and induces apoptosis and autophagy.These findings indicate that CRT0066101 is a promising small-mole-cule inhibitor for targeted HCC therapy with potential clinical applications.

Aim To investigate the inhibitory effect of the protein kinase D(PKD)-specific inhibitor CRT0066101 on hepatocellular carcinoma(HCC)and its underlying molecular mechanisms,providing new theoretical insights and therapeutic strategies for targe-ted HCC treatment.Methods HCC cell lines were treated with varying concentrations of CRT0066101.The inhibitory effects on cell proliferation were assessed using the CCK-8 assay,colony formation assay,and EdU staining.The impact on cell migration and inva-sion was evaluated through wound-healing assays and Transwell migration and invasion assays.was employed toThe effects of CRT0066101 on the phosphorylation levels of PKD and key proteins in the downstream PI3K/AKT signaling pathway were analyzed using Western blot.Additionally,the drug's regulatory effects on apoptosis and autophagy in HCC cells were examined using Western blot,flow cytometry,and the mRFP-GFP-LC3 dual-fluorescence reporter system.Results CRT0066101 significantly inhibited the pro-liferation,migration and invasion of HCC cells.West-ern blotting results demonstrated that CRT0066101 dose-dependently suppressed the phosphorylation of PKD family proteins and downregulated the activation of the PI3K/AKT signaling pathway.Furthermore,CRT0066101 treatment upregulated the expression of the pro-apoptotic protein Bax while downregulating the anti-apoptotic protein Bcl-2.It also markedly increased the expression levels of autophagy marker proteins Bec-lin-1 and LC3B-Ⅱ,suggesting that the drug simulta-neously induced apoptosis and autophagy in HCC cells.Conclusions CRT0066101 specifically inhibits PKD activity,blocks the PI3K/AKT signaling path-way,suppresses HCC cell proliferation and metastasis,and induces apoptosis and autophagy.These findings indicate that CRT0066101 is a promising small-mole-cule inhibitor for targeted HCC therapy with potential clinical applications.

OBJECTIVE: To investigate the mechanism of Radix Kansui (RK) stir-fried with vinegar (VRK) decreased hepatotoxicity in mice. METHODS: According to a random number table, 40 mice were randomly divided into negative control group (0.5% carboxymethylcellulose sodium, 20 mL/kg), positive control group (0.1% mixture of carbon tetrachloride in soybean oil, 20 mL/kg), RK group (the ethyl acetate extracts of RK, 250 g crude drug/kg) and VRK group (the ethyl acetate extracts of VRK, 250 g crude drug/kg) with 10 mice per group. All mice were administered orally by gavage daily for 7 continuous days. The morphology of liver tissues was examined to assess the liver injury by a transmission electron microscope. Hepatocyte apoptosis in vivo was determined by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nickend labeling (TUNEL) assay. Immunohistochemical technique was adopted to detect the expression of particular antiapoptotic and proapoptotic proteins in the mitochondrial pathways, including B-cell lymphoma (Bcl-2) and caspase-3, as well as the expression of inflammatory mediators, including nuclear factor kappa B (NF- κ B) and intercellular adhesion molecule-1 (ICAM-1). RESULTS: Liver injury and hepatocyte apoptosis were observed in RK mice, and the liver injury were significantly reduced in VRK-treated mice. In immunohistochemistry study, compared with the negative control group, RK inhibited dramatically the Bcl-2 protein expression and significantly increased the expression of caspase-3, NF- κ B and ICAM-1 (all P<0.01). Compared with the RK group, VRK group induced significant increase on Bcl-2 protein expression, and decreased the caspase-3, NF- κ B and ICAM-1 protein expression (P<0.05 or P<0.01). CONCLUSION The mechanism of reduced hepatotoxicity of VRK may be associated with the reduced inflammation, regulation of antiapoptotic and proapoptotic mediators in the mitochondrial pathway.

Objective:To determine the current status of multiple antibiotic residues in meat and meat products in Shanghai based on a 5-year surveillance， and perform the health risk assessment. Methods:We performed the examination in accordance with the Manual for National Food Contamination and Harmful Factor Risk Monitoring， and conduct health risk assessment according to the national limit standards on the monitoring data of 2016‒2020. Results:The total detection rate of multiple antibiotics in meat and meat products in Shanghai was determined to be 16.03%， in which the total unqualified rate was 1.97%. Moreover， the detection rate of quinolones was 2.78% and its unqualified rate was 0.83%. The unqualified rate of loxacin in cooked meat products was 2.12%. The detection rate of tetracyclines was 17.06% and its unqualified rate was 0.34%， in which the highest detection rate was identified in doxycycline （11.64%）. The detection rate of sulfonamides was 3.16%， in which the highest detection rate was in sulfamethazine （1.05%）. The detection rate of florfenicol was 5.15% and its unqualified rate was 0.12%. The difference of ofloxacin residues between diverse food categories （χ²=17.44， P<0.05） and processing links （χ² =14.10， P<0.05） was statistically significant. In addition， the sum amount of ofloxacin， enrofloxacin and ciprofloxacin in cooked meat products was higher than other food categories； the unqualified rate and residual amount of ofloxacin available in online stores and catering links were both higher. The residual amount of doxycycline and the unqualified rate in the online store link were significantly higher than those in other links. Based on preliminary assessment， the high exposure values in the 97.5 percentile of meat and meat products accounted for a very low proportion of the corresponding acceptable daily intake （ADI） and posed a low health risk to the population. Conclusion:The total detection rate of tetracyclines in meat and meat products is relatively high， which obviously accumulates in the offals of livestock and poultry. In addition， some antibiotics， such as ofloxacin and doxycycline， are relatively high in catering and online stores. It is recommended to strengthen the supervision of quinolones in cooked meat products， especially ofloxacin， enrofloxacin， and ciprofloxacin， and improve the supervision of doxycycline in meat and meat products in online stores.