Incorporating Network Pharmacology and Experimental Validation to Identify Bioactive Compounds and Potential Mechanisms of Digitalis in Treating Anaplastic Thyroid Cancer.

Anaplastic thyroid cancer (ATC) is one of the most lethal malignant tumors for which there is no effective treatment. There are an increasing number of studies on herbal medicine for treating malignant tumors, and the classic botanical medicine Digitalis and its active ingredients for treating heart failure and arrhythmias have been revealed to have significant antitumor efficacy against a wide range of malignant tumors. However, the main components of Digitalis and the molecular mechanisms of its anti-ATC effects have not been extensively studied. Here, we screened the main components and core targets of Digitalis and verified the relationship between the active components and targets through network pharmacology, molecular docking, and experimental validation. These experiments showed that the active ingredients of Digitalis inhibit ATC cell activity and lead to ATC cell death through the apoptotic pathway.

Alantolactone induces concurrent apoptosis and GSDME-dependent pyroptosis of anaplastic thyroid cancer through ROS mitochondria-dependent caspase pathway.

BACKGROUND: Anaplastic thyroid cancer (ATC) is one of the fatal cancers and has not effective treatments. Alantolactone (ATL), a terpenoid extracted from traditional Chinese medicinal herb Inula helenium L., confers significant anti-inflammatory, antibacterial and antitumor activity. However, the activity and mechanisms of ATL in ATC remain unclear. PURPOSE: To investigate the potential anti-ATC effects in vitro and in vivo and the mechanisms involved. METHODS: The anti-proliferative activity of Alantolactone (ATL) against ATC cells was analyzed through CCK-8 and colony formation assays. Flow cytometry assay was performed to assess the cell cycle, cell apoptosis, ROS, and mitochondrial membrane potential (ΔΨm), whereas the cellular localization of cytochrome c and calreticulin were determined using cellular immunofluorescence assays. The lactate dehydrogenase (LDH) enzyme activity in the cell culture medium was measured using a commercial LDH kit, whereas ELISA was conducted to assess the secretory function of IL-1ß. Western blot assays were conducted to determine the expression or regulation of proteins associated with apoptosis and pyroptosis. Subcutaneous tumor model of nude mice was established to evaluate the anticancer activity of ATL in vivo. The expression of Ki67, cyclin B1, cleaved-PARP, cleaved-caspase 3, and IL-1ß in the animal tumor tissues was profiled using immunohistochemistry analyses. RESULTS: Our data showed that ATL significantly inhibited the proliferation and colony formation activity of ATC cells. ATL induced ATC cell cycle arrest at G2/M phase, and downregulated the expression of cyclin B1 and CDC2. Furthermore, ATL induced concurrent apoptosis and pyroptosis in the ATC cells, and the cleavage of PARP and GSDME. It also significantly increased the release of LDH and IL-1ß. Mechanically, ATL-mediated increase in ROS suppressed the Bcl-2/Bax ratio, downregulated the mitochondrial membrane potential and increased the release of cytochrome c, leading to caspase 9 and caspase 3 cleavage. We also found that ATL induced the translocation of an immunogenic cell death marker (calreticulin) to the cell membrane. In addition, it inhibited the growth of the ATC subcutaneous xenograft model, and activated proteins associated with apoptosis and pyroptosis, with a high safety profile. CONCLUSION: Taken together, these results firstly demonstrated that ATL exerted an anti-ATC activity by inducing concurrent apoptosis and GSDME-dependent pyroptosis through ROS-mediated mitochondria-dependent caspase activation. Meanwhile, these cell deaths exhibited obvious characteristics of immunogenic cell death, which may synergistically increase the potential of cancer immunotherapy in ATC. Further studies are needed to explore deeper mechanisms for the anti- ATC activity of ATL.

Therapeutic efficacy of nutritional support by percutaneous endoscopic gastrostomy in critically ill patients: A self-control clinical trial.

BACKGROUND & OBJECTIVE: Percutaneous endoscopic gastrostomy (PEG) is a procedure to provide enteral nutrition for critically ill patients. It is commonly used in clinical practice; however, the widespread use of PEG is controversial. Our objective was to evaluate the therapeutic effect of nutritional support by PEG in these critically ill patients. METHODS: A total of 64 critically ill patients including 41 males and 23 females (aged 23-84) were identified by the Acute Physiology and Chronic Health Evaluation (APACHE) II scoring system during September 2004 to June 2012. The nutritional status before and after PEG was mainly assessed by the tricep skinfold thickness and serum albumin level. The nutritional status and pathological condition were assessed at 4, 8 and 12 weeks before and after PEG feeding. The assessment was according to the classical method of the human nutritional status. Follow-up was performed at one month, three months and 1.5 year after gastrostomy. Statistical analysis was performed by SPSS 11.5 software. The incidence of inhalation pneumonia and gastroesophageal regurgitation was compared by chi square (χ2) test. P<0.05 were considered statistically significant. RESULTS: Among the 64 patients, 9 patients died of their former diseases or related symptoms. Postoperative follow-up showed that both nutritional status and complications were improved after PEG in 55 patients (P<0.05). The serum albumin and tricep skinfold thickness levels were significantly increased. The incidence of hypoglycemia, hypocalcemia, hypokalemia and hyponatremia were lower than pre-operation. The frequencies of complications were significantly reduced. No severe complications occurred in any patient. CONCLUSIONS: Our study confirmed that PEG was a good long-term route of nutritional supply with no serious complications for critically ill patients.