Background: Cetuximab is extensively used in the treatment of metastatic colorectal cancer (mCRC). However, resistance poses a significant challenge to successful therapy. Recently, paraptosis, a non-classical programmed cell death, has garnered increased attention for its potential application value in antitumor treatments. We aimed to identify the essential pathways and signaling molecules involved in paraptosis inhibition and select them as therapeutic targets in cetuximab resistance. Additionally, engineered exosome technology is used as a drug delivery system with both targeted and effector properties. Results: By comparing the differential expression of paraptosis-related genes between drug-resistant colon cancer cells and sensitive cells, it was observed that the paraptosis level induced by cetuximab was significantly downregulated in drug-resistant cells. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified the focal adhesion kinase (FAK) signaling pathway as a key pathway involved in the suppression of paraptosis. The biological function of FAK in cetuximab-resistant cells was investigated through cell morphology observation, CCK-8 assay, colony formation assay, RT-qPCR, Western Blot, and loss-of-function experiments. The results showed that the FAK signaling pathway was significantly upregulated in cetuximab-resistant colon cancer cells, and siRNA interference targeting FAK could notably inhibit cell proliferation while upregulating the paraptosis level. Based on this, engineered colon cancer cells targeted and FAK siRNA loaded exosomes (CT-Exo-siFAK1) were constructed. In vitro experiments, CT-Exo-siFAK1 could effectively activate paraptosis and inhibit the proliferation of drug-resistant colon cancer cells. In vivo experiments also confirmed that CT-Exo-siFAK1 significantly suppressed tumor growth and metastasis while upregulating the paraptosis level. Conclusion: This study suggests that FAK signaling pathway-mediated inhibition of paraptosis levels is crucial in the sensitivity of cetuximab targeted therapy in colon cancer, and the use of engineered exosomes to deliver FAK siRNA may be an effective strategy to reverse cetuximab resistance.

Danhong injection (DHI) has been widely prescribed to patients with acute ischemic stroke (AIS). However, due to methodological deficiencies, previous research has not yet provided rigorous evidence to support the use of DHI in the treatment of AIS. Therefore, we designed this multicenter, randomized, controlled, and double-blind trial to evaluate the efficacy and safety of DHI for AIS. It is a randomized, multicenter, double-blind, placebo-controlled, adaptive clinical trial. A total of 864 eligible patients will be randomized into either the DHI or placebo group in a 2:1 ratio. All patients will be given the standard medical care as recommended by guidelines. Participants will undergo a 2-week treatment regimen and 76-day follow-up period. The primary outcome is the proportion of patients with a favorable outcome, defined as a score of 0–1 on the modified Rankin scale at day 90. Secondary outcomes include a change in the total score of the Chinese medicine symptom scales of “Xueyu Zheng” (blood stasis syndrome), the proportion of patients with a Barthel Index score of ≥90, the proportion of patients with an improvement in NIHSS score of ≥4 or NIHSS score of 0–1, quality of life measured by the EQ-5D scale, etc. Safety outcomes such as global disability (mRS ≥3) at day 90 will also be assessed. The changes in mRNA and microRNA profiles in 96 patients selected from certain centers will also be assessed. As this is an adaptive design, two interim analyses are prospectively planned, which will be carried out after one-third and two-thirds of patients have completed the trial, respectively. Based on the results of the interim analyses, the Data Monitoring Committee (DMC) will decide how to modify the study. This trial will provide high-quality evidence for DHI in treatment of AIS. Clinical Trials.gov NCT01677208 (Date of registration 22 December 2012).

Background: Bladder cancer (BC) is one of the most common cancers of the urinary system. Negative regulation of apoptotic pathways is of the most significant biological process in cancer. More accurate tumor characterization and stratification of BC patients for selection of more appropriate treatments are required. Methods: The data for this study are from the National Center for Biotechnology Information (NCBI)’s Online Mendelian Inheritance in Man (OMIM) database. Disease-associated genes were performed via multiple text-based Searching in Agilent Literature Search software version 3.2.2. MCODE version 1.32 was used for computational analysis of network for the gene complex detection. Genes with common biological processes or pathways were divided into the same module. DAVID was used for Gene ontology (GO) and pathway analysis. The OS time of hub gene expression was analyzed by GEPIA. The study used Pearson Correlation Coefficient for correlated calculation of the hub genes in the same module (Bladder Urothelial Carcinoma samples compared with normal samples). We enriched the modules and predict the regulated miRNAs by Cluepedia. Interactions within each pathway can be investigated and new potential associations are revealed through gene/miRNA enrichments. Results: A total of 187 BC-associated genes were got from OMIM and used for network construction. A total of seventy-five modules were found in the network. EGFR, AR, MET, RELA, TP53, TSG101 are hub genes (edges above 10) of the largest 3 modules. The results demonstrate that BC patients with low-expressed TSG101 have longer OS, and are associated with TP53. Low-expressed RELA and over-expressed AR patients have a higher survival time. Low-expressed TSG101 patients have a longer survival time. Conclusions: In our study, we found that miRNA17, miRNA20a, miRNA15a, has-let-7b and miRNA16 were miRNAs regulating the top 3 modules.