CCDC43 as a potential therapeutic target of Tian Yang Wan for the treatment of hepatocellular carcinoma by activating the hippo pathway.

Introduction: Hepatocellular carcinoma (HCC) prevalence is rising annually, but the existing treatment strategies are limited; therefore, it is crucial to explore new therapeutic approaches. Methods: Here, we investigate the potential anti-cancer mechanism of an herbal medicine called Tian Yang Wan (TYW) in the treatment of HCC. The relationship of CCDC43 with immunity and cell death was analyzed by bioinformatics. Confirming the tumor suppressor effect of TYW on HCC cells by proliferation, invasion, migration and apoptosis assays. Results: First, we analyzed by proteomics that CCDC43 expression was downregulated after TYW administration and promoted the hippo pathway. Then, a large sample's transcriptome study demonstrated that elevated CCDC43 expression was strongly correlated with clinical traits and a bad prognosis in HCC patients. Next, we observed through multiple advanced algorithms that CCDC43 is involved in a variety of oncology and immunology related pathways. Notably, we found higher tumor immune microenvironment with high CCDC43 expression. Furthermore, we demonstrated that CCDC43 is associated with immune checkpoints and found that it is a sensitive indicator of a large number of chemotherapeutic agents. Subsequently, we conducted experimental investigations to demonstrate the capacity of TYW to impede proliferation and migration, while inducing apoptosis in human HCC cell lines. Finally, we performed analysis of two cell death patterns which showed CCDC43 to be strongly correlated with multiple ferroptosis factors and cuproptosis factors. Discusion: In conclusion, our study comprehensively examined the prognostic, immunological, and therapeutic implications of CCDC43 in HCC, thereby elucidating the therapeutic mechanism of action in TYW.

An in silico-in vitro pipeline for drug cardiotoxicity screening identifies ionic pro-arrhythmia mechanisms.

BACKGROUND AND PURPOSE: Before advancing to clinical trials, new drugs are screened for their pro-arrhythmic potential using a method that is overly conservative and provides limited mechanistic insight. The shortcomings of this approach can lead to the mis-classification of beneficial drugs as pro-arrhythmic. EXPERIMENTAL APPROACH: An in silico-in vitro pipeline was developed to circumvent these shortcomings. A computational human induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM) model was used as part of a genetic algorithm to design experiments, specifically electrophysiological voltage clamp (VC) protocols, to identify which of several cardiac ion channels were blocked during in vitro drug studies. Such VC data, along with dynamically clamped action potentials (AP), were acquired from iPSC-CMs before and after treatment with a control solution or a low- (verapamil), intermediate- (cisapride or quinine) or high-risk (quinidine) drug. KEY RESULTS: Significant AP prolongation (a pro-arrhythmia marker) was seen in response to quinidine and quinine. The VC protocol identified block of IKr (a source of arrhythmias) by all strong IKr blockers, including cisapride, quinidine and quinine. The protocol also detected block of ICaL by verapamil and Ito by quinidine. Further demonstrating the power of the approach, the VC data uncovered a previously unidentified If block by quinine, which was confirmed with experiments using a HEK-293 expression system and automated patch-clamp. CONCLUSION AND IMPLICATIONS: We developed an in silico-in vitro pipeline that simultaneously identifies pro-arrhythmia risk and mechanism of ion channel-blocking drugs. The approach offers a new tool for evaluating cardiotoxicity during preclinical drug screening.

Comparison of structural characteristics and bioactivities of polysaccharides from loquat leaves prepared by different drying techniques.

In the present study, freeze drying, hot-air drying, vacuum drying, and microwave drying at the microwave powers of 400, 600, and 800 W, respectively, were utilized to dry loquat leaves for evaluating the effects of different drying techniques on the physicochemical structures and bioactivities of polysaccharides extracted from loquat leaves (LLPs). Results demonstrated that the physicochemical structures and bioactivities of LLPs significantly affected by different drying techniques. The degrees of esterification, molar ratios of constituent monosaccharides, contents of uronic acids, apparent viscosities, and molecular weights of LLPs were varied by different drying techniques. Additionally, LLPs, particularly LLP-M4 which extracted from loquat leaves prepared by microwave drying at the power of 400 W, exerted remarkable in vitro binding capacities, strong inhibitory effects on α-amylase and α-glucosidase, and obvious antioxidant activities. Results indicated that the microwave drying could be an efficient drying technique before extraction of bioactive LLPs, and LLPs had great potential applications in the functional food and pharmaceutical industries.

Structural characterization, antioxidant activity, and immunomodulatory activity of non-starch polysaccharides from Chuanminshen violaceum collected from different regions.

Chuanminshen violaceum has been used as an important traditional Chinese medicine and a popular tonic food in China. Polysaccharides are considered the major bioactive components in C. violaceum. In this study, in order well understand the chemical structures and bioactivities of non-starch polysaccharides in C. violaceum (CVPs), the physicochemical structures, antioxidant activities, and immunomodulatory activities of CVPs in C. violaceum collected from different regions of China were investigated and compared. Results showed that the constituent monosaccharides and Fourier transform infrared spectra of CVPs in C. violaceum collected from different regions were similar. However, their molar ratios of constituent monosaccharides, molecular weights, and contents of uronic acids were different. Furthermore, CVPs exerted remarkable antioxidant activities (ABTS and nitric oxide radical scavenging capacities) and immunomodulatory activities (promoted production of nitric oxide, IL-6, and TNF-α from RAW 264.7 macrophages in vitro). Meanwhile, the antioxidant and immunomodulatory activities of CVPs extracted from C. violaceum also varied by cultivated regions. Moreover, results indicated that the antioxidant and immunomodulatory activities of CVPs were closely correlated to their α-1,4-d-galactosiduronic linkages. Results are helpful for better understanding of the structure-bioactivity relationships of CVPs, and beneficial for the improvement of their applications in pharmaceutical and functional food industries.