Background: Intrahepatic cholangiocarcinoma (ICC) is an aggressive hepatobiliary malignancy characterized by a complex pathogenesis and poor prognosis. Telomere dysfunction and cellular senescence are involved in the pathogenesis of various types of cancers. However, the prognostic significance of telomere- and aging-related genes in ICC remains unclear. This study aimed to identify such genes with prognostic significance, and construct a prognostic risk model for ICC.
Methods: We screened prognostic genes associated with telomere-aging in patients using The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. We then integrated 10 base machine learning algorithms, and generated 101 model configurations via parameter optimization and algorithm combination strategies, to construct and validate a prognostic risk model. The clinical prognostic value was analyzed using a nomogram. Immune infiltration, drug sensitivity, immune responses, and subgroups were analyzed based on telomere-aging-related prognostic genes. We also validated core gene expression in cell lines and tissue samples using the quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and enrolled 76 patients with ICC to identify the prognostic value of key genes.
Results: We constructed a prognostic model using the telomere-aging-related prognostic genes BET1L, RAD50, ANXA1, and AURKA. Survival analysis revealed a significant difference in overall survival between high- and low-risk groups. The expression of these genes was significantly increased in ICC cell lines and tissues. High BET1L expression was significantly associated with lymph node metastasis, tumor-node-metastasis (TNM) stage, tumor differentiation, and a poor prognosis for patients with ICC. Knocking down BET1L significantly reduced the proliferative ability of HUCCT1 cells.
Conclusions: We established a risk model comprising the telomere-aging-related prognostic genes BET1L, RAD50, ANXA1, and AURKA to predict the prognosis of patients with ICC. Elevated BET1L expression indicated a poor prognosis for patients with ICC, and low BET1L expression inhibited HUCCT1 cell proliferation.
Colorimetric assays offer a low-cost, accessible means of diagnostic testing but often suffer from subjective interpretation and variability caused by inconsistent imaging conditions. To address these challenges, we present HueTools, a comprehensive image processing software that enables quantitative development of paper-based colorimetric assays using smartphone imaging. HueTools integrates a mobile app, and a web platform to standardize image capture, apply precise color correction, and predictive modeling of analyte concentration using interpretable machine learning. The system supports a complete workflow: from image acquisition and color calibration to region-of-interest (ROI) selection, signal extraction, and statistical analysis. The system enables perceptually grounded color analysis using the CIELAB space and provides quantitative metrics, including LoB, LoD, and LoQ via interpretable ensemble-based customization models. HueTools was validated using a lateral flow dipstick luteinizing hormone (LH) and vertical-flow alanine transaminase (ALT) assays. The results demonstrate HueTools' ability to reduce human error, improve assay reproducibility, and provide feedback for optimizing assay design. It also supports seamless transitions between field testing and lab analysis, allowing researchers to capture images on-site and perform in-depth analysis remotely. HueTools offers a hardware-independent, cloud-based solution for assay developers, streamlining workflows while minimizing costs associated with dedicated readers. Its accessibility, automation, and cross-platform compatibility make it well-suited for research and development of colorimetric point-of-care diagnostics, especially in resource-limited settings.
Background: This study developed and systematically optimized an ion-pair hydrophilic interaction liquid chromatography (IP-HILIC) system for oligonucleotide medicine analysis, aiming to the issues of insufficient resolution and prolonged retention times commonly encountered in traditional hydrophilic interaction liquid chromatography (HILIC) and ion-pair reversed-phase liquid chromatography (IP-RPLC). Therefore, this study aimed to optimize and validate a TEAA-based IP-HILIC analytical system for both the effective separation and quantitative analysis of oligonucleotides, including evaluations of linearity, sensitivity, precision, and robustness.
Methods: By synthesizing an amino/hexadecyl-bifunctionalized silica matrix and systematically optimizing mobile phase parameters (including ion-pair reagent type and concentration, organic solvent, and buffer pH), a highly efficient separation system was established. To validate the separation efficiency, a 21-mer oligonucleotide Fomivirsen and its truncated impurities (5'-21-1 and 3'-21-1) were synthesized, along with Food and Drug Administration(FDA)-approved Kynamro (20-mer), Macugen (28-mer), and Exondys 51 (30-mer) for method evaluation.
Results: Experimental results demonstrated that under pH 8.0 conditions, a mobile phase system composed of 50 mM triethylamine acetate (TEAA) and acetonitrile enabled baseline separation of 20-30-mer oligonucleotides with a resolution ≥1.5. Using Fomivirsen impurities (e.g., 5'-21-1 and 5'-21-3) as examples, the retention times were optimized to between 3.76 and 6.76 minutes, achieving a resolution of 3.82. Acetonitrile shortened the analysis time by 34% compared with methanol, while 50 mM TEAA maintained resolution in the range of 2.58-3.71 and avoided the excessive retention seen at 100 mM, where the peak time extended to 7.57 minutes.Method validation demonstrated that the method exhibited good linearity (R²>0.999) over the concentration range of 0.05-10 μg/mL, with a limit of detection (LOD) of 0.2 μM and a limit of quantitation (LOQ) of 0.6 μM. The method also showed satisfactory precision (intra-day RSD < 1.5%, inter-day RSD < 2.1%) and was robust against minor variations in mobile phase composition, column temperature, and flow rate. Compared to traditional methods, IP-HILIC demonstrated significantly superior resolution (1.50-2.09) for long-chain oligonucleotides (e.g., 30-mer Exondys 51) over IP-RPLC (1.30) and HILIC (1.20), with better peak symmetry (tailing factor <1.2).
Conclusion: This study provides an systematically optimized and validated analytical platform for the quality control of oligonucleotide medicines, particularly showing significant advantages in the separation of key impurities (e.g., n-1 sequences) for long-chain oligonucleotides (20-30 mer), laying a technical foundation for the precise analysis of hydrophilic biopolymers.
In the automation of life science experiments, verifying and recording the types, positions, and states of labware are essential for quality assurance and traceability of experimental results. To achieve this, researchers often record static images or videos within lab automation instruments and workstations. However, the absence of specialized open-source software for labware detection significantly hinders the efficient utilization of these images and videos in automated life science experiments. To address this issue, we developed OT2Eye, a labware detection tool tailored for the images and videos of the Opentrons OT-2, an affordable liquid-handling robot. OT2Eye demonstrated high accuracy in identifying various labware types, including microwell plates and tip racks, and precisely locating them within the OT-2 workspace. Additionally, OT2Eye accurately detected the presence or absence of tips on tip racks. OT2Eye renders labware states machine-readable, streamlining labware status management and logging, and enabling integration with AI-driven laboratory automation systems using cost-effective automated pipetting robots.
Different biochemical assays yield different rates of false positives than others either due to the nature of the enzyme, the technology associated with the bioassay, or properties of the compounds being screened. Ensuring that the right counter-screens are in place to identify false positives without wasting time and resources on them is of great importance. Herein we describe the results of a high throughput screen (HTS) against non-structural protein 3 (nsp3) protease PLPro, which resulted exclusively in false-positive hits. By triaging hit compounds through purification of metal chelating resin, we identified contamination by either copper or palladium as the most likely source of false positives from the library screening campaign. We then performed a systematic assessment of the vulnerability of nsp3 protease screening to metal contamination and evaluated common additives to combat the inhibitory effects of different metal salts. We further conducted a thorough survey of the literature reports of nsp3 HTS campaigns with a focus on the presence of additives and what metal susceptibility was likely, given the results of our work. We conclude that the majority of reported nsp3 screens are susceptible to copper contamination with a smaller proportion also potentially susceptible to palladium contamination.
Solvent evaporation, a ubiquitous yet often limiting step in chemical synthesis, routinely creates critical bottlenecks in research workflows. We introduce a novel benchtop solvent evaporation platform engineered to dramatically accelerate this process within a standard fume hood. Designed with flexible hardware and software architecture, this system is primed for autonomous control via Python scripting, transitioning from its current manual operation to fully automated capabilities. The platform offers two versatile configurations: a single-container evaporator for individual sample precision and a batch evaporator for high-throughput, simultaneous processing of multiple vials. Both variants feature a custom-designed, chemically inert 3D-printed adapter, equipped with precise compressed nitrogen and air inlet ports, alongside an efficient exhaust. Rapid and uniform solvent removal is achieved through a synergistic approach: integrated heating pads maintain optimal temperature in the vial block, while pressurized nitrogen is directed through uniquely helical channels within the adapter, generating a powerful vortex that significantly accelerates evaporation. Rigorous studies confirm the platform's robust and uniform solvent removal performance, conclusively demonstrating its potential to fundamentally alleviate key bottlenecks in chemical synthesis, thereby enhancing productivity and accelerating discovery. This innovative system represents a vital step towards fully automated and efficient chemical laboratories.

