Cell viability and cytotoxicity assays play a crucial role in drug screening and evaluating the cytotoxic effects of various chemicals. The quantification of cell viability and proliferation serves as the cornerstone for numerous in vitro assays that assess cellular responses to external factors. In the last decade, several studies have developed guidelines for defining and interpreting cell viability and cytotoxicity based on morphological, biochemical, and functional perspectives. As this domain continues to experience ongoing growth, revealing new mechanisms orchestrating diverse cell cytotoxicity pathways, we suggest a revised classification for multiple assays employed in evaluating cell viability and cell death. This classification is rooted in the cellular compartment and/or biochemical element involved, with a specific focus on mechanistic and essential aspects of the process. The assays are founded on diverse cell functions, encompassing metabolic activity, enzyme activity, cell membrane permeability and integrity, ATP content, cell adherence, reduction equivalents, dye inclusion or exclusion, constitutive protease activity, colony formation, DNA fragmentation and nuclear splitting. These assays present straightforward, reliable, sensitive, reproducible, cost-effective, and high-throughput approaches for appraising the effects of newly formulated chemotherapeutic biomolecules on the cell survival during the drug development process.

Conventional treatment methods are not effective enough to fight the rapid increase in cancer cases. The interest is increasing in the investigation of herbal sources for the development of new anticancer therapeutics. Particularly, much attention is given to finding combined phytochemical/chemotherapeutic treatment models to overcome drug resistance and decrease side effects. The aim was to investigate the antitumor capacity of Hypericum alpestre herb extract in vitro and in vivo, either alone or combined with the inhibitors of the L-arginine/polyamine/nitric oxide pathway and characterize its active phytochemicals using advanced chromatographic techniques. The antioxidant capacity of H. alpestre extract was assessed through chemical spectrophotometric tests (DPPH and ABTS) and in biological systems using Cellular Antioxidant Activity assay. The inhibitory effect of H. alpestre extract on the growth of human colorectal (HT29) and breast cancer (MCF-7) cell cultures was explored by the MTT test. The genotoxicity of the tested extract was studied using a comet assay. In vivo, the antitumor properties of H. alpestre and its combinations were explored in a rat mammary gland carcinogenesis model induced by subcutaneous injection of 7,12-dimethylbenz[a]anthracene. The polyphenolic substances present in H. alpestre extract have been characterized using the LC-Q-Orbitrap HRMS system. The H. alpestre extract expressed promising antiproliferative effects on MCF-7 and HT29 cells. The extract did not exhibit genotoxic activity nor possessed antigenotoxic properties. The in vivo rat mammary carcinogenesis model data showed that the H. alpestre extract stimulated the activity of antioxidant enzymes in the liver, brain, and tumors of rats in the experimental groups, demonstrating its antioxidant protective effects. The herb alone and in combination with N ω-OH-nor-L-arginine and N ω-nitro-L-arginine methyl ester exhibited pro-/antioxidant, antiproliferative, anti-angiogenic, and cytotoxic effects.

Langerhans cells(LCs) play a critical role in skin immune responses and the development of psoriasis. Yinxieling(YXL) is a representative herbal medicine formula for the treatment of psoriasis in South China. It was confirmed to improve psoriasis without obvious side effects in the clinic. Here we attempted to clarify whether and how YXL regulates the differentiation and functions of LCs in Imiquimod(IMQ)-induced psoriasis-like C57BL/6 mouse model in vivo, and induced LCs in vitro. The Psoriasis Area Severity Index (PASI) score was used to evaluate efficacy of YXL for IMQ-induced psoriasis-like mice. Flow cytometry was utilized to analyze the effects of YXL to regulate the differentiation, migration, mature and antigen presentation of LCs. The results show that YXL significantly alleviated skin inflammation, as reduced in PASI score and classic psoriatic characteristics in pathological sections. Although there was not any effect on the proportion of total DCs in the skin draining lymph nodes, the expression of epidermal LCs and its transcription factor PU.1 was significantly inhibited, and its migration to draining lymph nodes and maturation were also inhibited. Further data showed that the number of antigen-carrying LC in the epidermis increased, indicating that YXL could effectively inhibit the antigen presentation of LCs. YXL also significantly inhibited the differentiation of LC in vitro. Further data showed that YXL decreaded the relatve expression of TGFβmRNA and IL-23 mRNA. Thus, YXL allievates psoriasis by regualting differentiation, migration, maturation and antigen presentation via TGFβ/ PU.1/IL-23 signal axis.

Non-coding RNA species play important roles in biological mechanisms that regulate glioma initiation and progression. Recently, evidence suggests that ZNFX1 antisense RNA 1 ( ZFAS1) has the ability to act as an oncogene or tumour suppressor, and so plays critical regulatory functions in the development and progression of many types of cancers such as lung, renal and hepatocarcinoma. The roles of ZFAS1 in glioma cancer are still unclear, and there are numerous potential pathways to explore. The current work provides a scoping review of studies on ZFAS1’s functions and underlying molecular mechanisms in the initiation and progression of glioma, as well as a possible field of research to be examined further. A literature search was carried out using Scopus, PubMed, and Web of Science (WoS) using a specified search string, and the data gathered was discussed and reported. This scoping review comprised five original research papers that study ZFAS1 and its roles in gliomagenesis. ZFAS1 was found to be highly upregulated in glioma. Tumour-node-metastasis (TNM) stage, lymph node metastases, and overall survival were revealed to be significantly associated with ZFAS1 status and regulated via several pathways and interactions, such as miRNA signalling, Epithelial-to-Mesenchymal Transition (EMT) and Notch signalling pathway. Furthermore, ZFAS1 knockdown decreased cell proliferation, migration, and invasion while promoting cell death, implying that ZFAS1 is involved in the glioma cancer progression. The evaluation of their diagnostic importance and therapeutic potential may aid in the development of novel therapies for glioma cancer.

As the field of stem cell-based therapies continues to evolve, the ability to monitor the regenerative capacity of stem cell following delivery to target tissues has become increasingly relevant. While Bromodeoxyuridine (BrdU) and green-fluorescent protein (GFP) labeling techniques are often used, limited knowledge exists regarding their effects on the regeneration capacity of stem cells. In order to further understand the impact of these techniques in a clinical scenario, porcine adipose stem cells (ASCs) were derived, cultured and subsequently labeled with BrdU or GFP using transfection (GFP-t) or infection (GFP-i) by lentivirus, followed by investigation of downstream labeling effects on ASCs proliferation, differentiation, colony formation, secretion of regenerative cytokines, growth factors and healing ability. Neither BrdU nor GFP labeling led to gross morphological changes in adipose stem cells. Both groups of GFP labeled cells maintained a high percentage of signal intensity for at least 28 days, with a decrease in cell proliferation ability in the GFP-i group. In contrast, BrdU labelling began displaying weaker signal strength after 7-days of experiments. Both tracers negatively affected osteogenic and adipogenic differentiation of ASCs, causing decrease in the secretion of the regeneration markers VEGF and TNF-α. In contrast, in-vitro wound healing ability of labeled ASCs and secretion of IL-8, IL-10, and TGF-β was not affected by either method. Our results demonstrate that BrdU labelling may be more effective for short-term in-vitro studies due to the relative ease of preparation and shorter lifespan of the tracer signal. Contrarily, for studies with a longer duration, GFP labeling of cells by transfection provides more durable and stable signal. While further studies are needed, our results indicate that labelling mode can alter regeneration capacity of stem cells and should, therefore, be optimized prior to clinical translation experiments.