Ovatodiolide inhibited hepatocellular carcinoma stemness through SP1/MTDH/STAT3 signaling pathway.

Hepatocellular carcinoma (HCC) is characterized with high recurrence and mortality, and the clinical treatments for HCC are very limited. Hepatocellular carcinoma stem cells are the root of HCC progress, recurrence, and multidrug resistance. Ovatodiolide (OVA) is a bioactive diterpenoid served as an inflammatory and immunotherapeutic responses modulator. In this research, we found OVA inhibited HCC stemness through inhibiting MTDH gene transcription. Moreover, we firstly discovered transcription factor SP1 bound to the promoter region of MTDH to transcriptionally regulate MTDH level. Mechanically, we demonstrated OVA decreased SP1 protein stability to transcriptionally inhibit MTDH gene, and inhibited the nuclear translocation of p65, and then diminished IL-6 level to suppress JAK/STAT3 signaling pathway, eventually decreases CD133 level and the stemness of HCC. Furthermore, we demonstrated ACT004, OVA derivative with high metabolic stability towards cytochrome P450 enzymes, showed no genotoxicity and no accumulative or delayed toxicities after long-term administration in rats. And the in vivo efficacy experiments indicated ACT004 inhibited tumor growth of hepatocellular carcinoma. In conclusion, we revealed the mechanism of OVA in regulating HCC stemness, detected the toxicity of OVA derivative and evaluated the in vivo efficacy which lays a foundation for further discovery of anti-HCC stem cell agents and provide a new strategy for the application of OVA in clinical treatment.

Preclinical safety evaluation of a bivalent inactivated EV71-CA16 vaccine in mice immunized intradermally.

Hand, foot and mouth disease is a common acute viral infectious disease that poses a serious threat to the life and health of young children. With the development of an effective inactivated EV71 vaccine, CA16 has become the main pathogen causing HFMD. Effective and safe vaccines against this disease are urgently needed. In our previous study, a bivalent inactivated vaccine was shown to have good immunogenicity and to induce neutralizing antibodies in mice and monkeys. Repeated administration toxicity is a critical safety test in the preclinical evaluation of vaccines. In this study, BALB/c mice were used to evaluate the toxicity of the bivalent vaccine after multiple intradermal administrations. Clinical observation was performed daily, and body weight, food intake, hematological characteristics, serum biochemical parameters, antinuclear antibodies, CD4+/CD8a+ T-cell proportions, bone marrow smear results and pathology results were recorded. The results showed that there was no significant change at the injection site and no adverse reactions related to the vaccine. The bivalent inactivated EV71-CA16 vaccine exhibits good safety in mice, and these results provide a sufficient basis for further clinical trials.

Consistency analysis of high-sensitivity cardiac troponin I in peripheral blood and venous blood by quantum dot immunofluorescence assay and clinical application in acute myocardial infarction.

Background: Troponin is an important marker for the diagnosis of acute myocardial infarction (AMI). The detection of troponin in peripheral blood is simpler and more convenient than that in venous blood, which has attracted more and more clinical attention. The purpose of this study is to establish a novel method for the rapid detection of high-sensitivity troponin I (hs-cTnI) in peripheral blood by quantum dot fluorescence immunoassay and evaluated the clinical accuracy of the method. Methods: A total of 90 patients with chest pain admitted to Wuxi Second People's Hospital of Nanjing Medical University had peripheral blood and venous blood samples collected for detection of hs-cTnI by rapid quantum dot fluorescence immunoassay. The differences between the two methods were evaluated, as well as the analytical performance and clinical diagnostic efficacy of hs-cTnI detection by quantum dot fluorescence immunoassay. The final diagnosis was determined by two independent cardiologists. Results: This study verified the precision, linear range and sensitivity of the novel detection method. There was good correlation between the results of hs-cTnI quantum dot fluorescence immunoassay for peripheral blood and the results for venous blood (regression equation Y=1.026x+0.521, R2=0.9337); 94.4% (85/90) of the data were within the conformance limit. In addition, in the analysis of 52 patients with confirmed AMI, the clinical specificity of the quantum dot fluorescence immunoassay in peripheral blood was the same as that in venous blood samples (89.5%:89.5%). Finally, the area under the receiver operating characteristic (ROC) curve of the peripheral blood quantum dot fluorescence immunoassay was 0.9352, the 95% confidence interval (CI) was 0.8829 to 0.9876, the cut-off value was 1.598, and the sensitivity was 82.69%, which was not significantly different from the venous blood method (P value =0.089). Conclusions: Rapid detection of hs-cTnI by quantum dot fluorescence immunoassay in peripheral blood is feasible. It has a high correlation and consistency with the venous blood method, as well as a high clinical diagnostic value for AMI and is more convenient and easier to detect.

Preclinical safety assessment of a combined vaccine against Hepatitis a virus and enterovirus 71.

Since 2007, Hepatitis A (HAV) vaccination has been a part of the National Immunization Program of China. Recognizing enterovirus 71 (EV71) as the most important pathogen in severe hand, foot and mouth disease, an inactivated EV71 vaccine was successfully marketed in 2015. Based on the concept of one vaccine preventing two diseases and owing to similarities in vaccine preparation and the overlap of the eligible population, a combination of the inactivated HAV vaccine and inactivated EV71 vaccine is theoretically feasible and desirable. However, the optimal vaccinationschedule for this combination vaccine has yet to be optimized. Use of this combined vaccine would not only decrease the number of vaccinations, but also lower associated cost. This study aimed to investigate the toxicity and adverse reactions of the combined HAV-EV71 vaccine under Good Laboratory Practice conditions to provide a reference for clinical studies/applications in the future. CD®(Sprague Dawley) IGS rats were employed for single-dose toxicity testing using a high dose, and repeated-dose toxicity testing using high, as well as low doses. Animals that received only a single dose showed no obvious clinical symptoms nor abnormal body weight, and no significant gross pathological change at the experimental endpoint at necropsy. In the rats injected with three doses, phagocytosis of basophilic granules by macrophages was observed in the inguinal, mesenteric, and local lymph nodes, besides irritation at the administration site. At 56 days after the last dose, no significant histopathological change was observed in the lymph nodes, and local irritation gradually faded. Further, systematic allergy testing was performed in guinea pigs. After systemic sensitization and challenge with the HAV-EV71 vaccine, animals showed normal weight gain and no allergic reactions. This study, therefore, confirmed a good safety profile of the inactivated HAV and EV71 combined vaccine.

Self-assembling peptides improve the stability of glucagon-like peptide-1 by forming a stable and sustained complex.

The multiple physiological characterizations of glucagon-like peptide-1 (GLP-1) make it a promising drug candidate for the treatment of T2DM. However, the short half-life of GLP-1 limits its clinical utility. Self-assembling peptides are presumed to wrap GLP-1 peptide, and this helps to prolong the stability of GLP-1 consequently. The aim of this study was to investigate whether self-assembling peptides could be applied to prolong the half-life of GLP-1 as sustained release preparations. In this study, five different self-assembling peptides were employed. The formation of the complexes was monitored using gel filtration and mass spectrometry and was simulated by Molecular Dynamics. Stabilization, insulin secretion stimulation, and glucose tolerance tests were performed to investigate the physiological characteristics retained by GLP-1 following complex formation with self-assembling peptides. Our findings revealed that, among the five different self-assembling peptides tested, complex of Pep-1 and GLP-1 exhibited a remarkable extension in the half-life of GLP-1. In addition, the experimental animals treated with a GLP-1/Pep-1 complex exhibited better blood glucose clearance activity over a greater duration of time than the animals treated with GLP-1 alone. Based on our results, an adjustment of the Pep-1 and GLP-1 ratios is presumed to be able to control the half-life of GLP-1 (e.g., medium-acting and long-acting). Collectively, the findings in this study suggest that the self-assembling peptide Pep-1 could serve as a powerful drug preparation tool to extend the short half-life of therapeutic peptides.