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Malignant pleural effusion (MPE), the presence of malignant cells in pleural fluid, is often the first sign of many cancers and occurs in patients with metastatic malignancies. Accurate detection of tumor cells in pleural fluid is crucial because the presence of MPE denotes an advanced stage of disease and directs a switch in clinical managements. Cytology, as a traditional diagnostic tool, has limited sensitivity especially when tumor cells are not abundant, and may be confounded by reactive mesothelial cells in the pleural fluid. We describe a highly sensitive approach for rapid detection of metabolically active tumor cells in MPE via exploiting the altered glucose metabolism of tumor cells relative to benign cells. Metabolically active tumor cells with high glucose uptake, as evaluated by a fluorescent glucose analog (2-NBDG), are identified by high-throughput fluorescence screening within a chip containing 200,000 addressable microwells and collected for malignancy confirmation via single-cell sequencing. We demonstrate the utility of this approach through analyzing MPE from a cohort of lung cancer patients. Most candidate tumor cells identified are confirmed to harbor the same driver oncogenes as their primary lesions. In some patients, emergence of secondary mutations that mediate acquired resistance to ongoing targeted therapies is also detected before resistance is manifested in the clinical imaging. The detection scheme can be extended to analyze peripheral blood samples. Our approach may serve as a valuable complement to cytology in MPE diagnosis, helping identify the driver oncogenes and resistance-leading mutations for targeted therapies.

The sweet-mellow taste sensation is a unique and typical feature of premium congou black tea infusions. To explore the key taste-active compounds that influence the sweet-mellow taste, a sensory and molecular characterization was performed on thirty-three congou black tea infusions presenting different taste qualities, including the sweet-mellow, mellow-pure, or less-mellow taste. An integrated application of quantitative analysis of 48 taste-active compounds, taste contribution analysis, and further validation by taste supplementation experiments, combined with human sensory evaluation revealed that caffeine, γ-aminobutyric acid, rutin, succinic acid, citric acid, and gallic acid negatively affect the sweet-mellow taste, whereas glucose, sucrose, and ornithine positively contribute to the sweet-mellow taste of congou black tea infusions. Particularly, rutin, γ-aminobutyric acid, gallic acid, and caffeine, which impart the major inhibitory effect to the manifestation of the sweet-mellow taste, were identified as the key influencing components through stepwise screening and validation experiments. A modest level of these compounds was found to be favorable for the development and manifestation of the sweet-mellow taste. These compounds might potentially serve as the regulatory targets for oriented-manufacturing of high-quality sweet-mellow congou black tea.

Genetic and transcriptional profiling, as well as surface marker identification of single circulating tumor cells (CTCs) have been demonstrated. However, quantitatively profiling of functional proteins at single CTC resolution has not yet been achieved, owing to the limited purity of the isolated CTC populations and a lack of single-cell proteomic approaches to handle and analyze rare CTCs. Here, we develop an integrated microfluidic system specifically designed for streamlining isolation, purification and single-cell secretomic profiling of CTCs from whole blood. Key to this platform is the use of photocleavable ssDNA-encoded antibody conjugates to enable a highly purified CTC population with <75 ‘contaminated’ blood cells. An enhanced poly-L-lysine barcode pattern is created on the single-cell barcode chip for efficient capture rare CTC cells in microchambers for subsequent secreted protein profiling. This system was extensively evaluated and optimized with EpCAM-positive HCT116 cells seeded into whole blood. Patient blood samples were employed to assess the utility of the system for isolation, purification and single-cell secretion profiling of CTCs. The CTCs present in patient blood samples exhibit highly heterogeneous secretion profile of IL-8 and VEGF. The numbers of secreting CTCs are found not in accordance with CTC enumeration based on immunostaining in the parallel experiments.

Aroma is an indispensable factor that substantially impacts the quality assessment of black tea. This study aims to uncover the dynamic alterations in the sweet and floral aroma black tea (SFABT) throughout various manufacturing stages using a comprehensive analytical approach integrating gas chromatography electronic nose, gas chromatography–ion mobility spectrometry (GC-IMS), and gas chromatography–mass spectrometry (GC-MS). Notable alterations in volatile components were discerned during processing, predominantly during the rolling stage. A total of 59 typical volatile compounds were identified through GC-IMS, whereas 106 volatile components were recognized via GC-MS throughout the entire manufacturing process. Among them, 14 volatile compounds, such as linalool, β-ionone, dimethyl sulfide, and 1-octen-3-ol, stood out as characteristic components responsible for SFABT with relative odor activity values exceeding one. This study serves as an invaluable theoretical platform for strategic controllable processing of superior-quality black tea.

Background Persistent infection with high-risk human papillomavirus (HR-HPV) is the primary cause of cervical cancer. Understanding genotype distribution and evaluating screening strategies are essential for effective prevention. Methods We retrospectively analyzed cervical cancer screening data from 97,686 women aged 35–64 years in Nanjing (2021–2023). Among these, 30,147 underwent combined cytology and HPV testing and 67,539 TCT alone. HPV genotyping was further performed in 3,362 histologically confirmed cervical cancer cases (3,014 squamous cell carcinomas [SCC] and 348 adenocarcinomas [ADC]) from multiple regions in China. Results Combined screening achieved a significantly higher detection rate of abnormalities than TCT alone (13.70% vs. 1.79%, p  < 0.001). Overall HPV positivity was 11.19%, increasing with age and peaking at 16.85% in women aged 60–64. The most frequent genotypes were HPV52, HPV58, and HPV16. In cervical cancer cases, HPV was detected in 92.73% of SCC and 59.77% of ADC. The proportion of HPV-negative cancers increased with age, particularly in ADC. Conclusions Combined TCT and HPV testing improves detection of cervical lesions compared with cytology alone. The observed age-specific and histology-specific differences in HPV prevalence and genotype distribution emphasize the need for tailored screening strategies, particularly for older women. These findings provide region-specific evidence to support the refinement of cervical cancer prevention and control strategies in China, particularly in contexts with similar demographic and epidemiological characteristics to the study population.

Background Cervical cancer (CC) is the second leading cause of cancer death among women worldwide. Epigenetic regulation of gene expression through DNA methylation and hydroxymethylation plays a pivotal role during tumorigenesis. In this study, to analyze the epigenomic landscape and identify potential biomarkers for CCs, we selected a series of samples from normal to cervical intra‐epithelial neoplasia (CINs) to CCs and performed an integrative analysis of whole‐genome bisulfite sequencing (WGBS‐seq), oxidative WGBS, RNA‐seq, and external histone modifications profiling data. Results In the development and progression of CC, there were genome‐wide hypo‐methylation and hypo‐hydroxymethylation, accompanied by local hyper‐methylation and hyper‐hydroxymethylation. Hydroxymethylation prefers to distribute in the CpG islands and CpG shores, as displayed a trend of gradual decline from health to CIN2, while a trend of increase from CIN3 to CC. The differentially methylated and hydroxymethylated region‐associated genes both enriched in Hippo and other cancer‐related signaling pathways that drive cervical carcinogenesis. Furthermore, we identified eight novel differentially methylated/hydroxymethylated‐associated genes (DES, MAL, MTIF2, PIP5K1A, RPS6KA6, ANGEL2, MPP, and PAPSS2) significantly correlated with the overall survival of CC. In addition, no any correlation was observed between methylation or hydroxymethylation levels and somatic copy number variations in CINs and CCs. Conclusion Our current study systematically delineates the map of methylome and hydroxymethylome from CINs to CC, and some differentially methylated/hydroxymethylated‐associated genes can be used as the potential epigenetic biomarkers in CC prognosis. The differentially DMR/DhMR occurred earlier and much more than that of CNVs which would be more suitable for a marker to detect the disease progression. The differentially DMR/DhMR‐associated genes enrich in Hippo and other cancer‐related signaling pathways. Eight DMR/DhMR‐associated genes significantly correlate with the overall survival of CC.

We report an easily setup, reliable and automatic microfluidic sample transfer and introduction system. Two different function liquid detection modules were developed to separately perform rapidly removing of a large approximate volume of air off chip and a low-speed high precision small volume of air purging process on chip incorporating liquid-on-chip handling module. As a proof of concept, we demonstrated that a small volume of radioactive sample as low as 5 μL could be successfully transferred and introduced from vials to the desired location in the microfluidic chip with minimal loss (2.1 ± 0.4 %, n  = 3). The total time of the sample transfer and introduction was less than 1 min. The complete automation would facilitate the safe handling of the dangerous and toxic materials, such as radioactive compound.

SLC7A11, a plasma membrane protein, has been implicated as an oncogene in various cancers, including hepatocellular carcinoma (HCC). Its role in HCC pathogenesis, particularly in relation to ferroptosis, is not well understood. This study aims to investigate the function of SLC7A11 with ferroptosis and its interaction in development of HCC. Clinical HCC tissue samples were used to analyze the expression of SLC7A11 by RT-PCR. The impact of SLC7A11 on HCC cell viability, proliferation, and migration was assessed by CCK-8, AlamarBlue and Transwell. Protein-protein interactions were explored using co-immunoprecipitation and immunofluorescence. The effect of SLC7A11 on ferroptosis was evaluated by iron levels, ROS, and GSH. The impact of sorafenib and doxorubicin (DOX) on HCC cells was analyzed using cell viability assay. SLC7A11 was found to be highly expressed in HCC tissues and was correlated with tumor size and poor prognosis. Overexpression of SLC7A11 in HCC cells promoted cell viability, proliferation, and migration. Additionally, SLC7A11 overexpression mitigated erastin-induced ferroptosis, as evidenced by decreased ROS levels and increased GSH levels. We also discovered that SLC7A11 interacted with HSPB1. HSPB1 inhibited erastin-induced ferroptosis. Furthermore, a portion of the cell death induced by sorafenib and DOX is attributed to ferroptosis, with HSPB1 and SLC7A11 inhibiting the death induced by the two drugs, respectively. SLC7A11 plays a significant role in HCC progression by inhibiting ferroptosis, and its interaction with HSPB1 is a critical pathway in this process. Targeting the SLC7A11-HSPB1 axis may provide a novel therapeutic strategy for HCC treatment, highlighting the importance of understanding the mechanisms of ferroptosis in cancer cells.

Persistent infection with high-risk human papillomavirus (HR-HPV) is the primary cause of cervical cancer. Understanding genotype distribution and evaluating screening strategies are essential for effective prevention. We retrospectively analyzed cervical cancer screening data from 97,686 women aged 35-64 years in Nanjing (2021-2023). Among these, 30,147 underwent combined cytology and HPV testing and 67,539 TCT alone. HPV genotyping was further performed in 3,362 histologically confirmed cervical cancer cases (3,014 squamous cell carcinomas [SCC] and 348 adenocarcinomas [ADC]) from multiple regions in China. Combined screening achieved a significantly higher detection rate of abnormalities than TCT alone (13.70% vs. 1.79%, p < 0.001). Overall HPV positivity was 11.19%, increasing with age and peaking at 16.85% in women aged 60-64. The most frequent genotypes were HPV52, HPV58, and HPV16. In cervical cancer cases, HPV was detected in 92.73% of SCC and 59.77% of ADC. The proportion of HPV-negative cancers increased with age, particularly in ADC. Combined TCT and HPV testing improves detection of cervical lesions compared with cytology alone. The observed age-specific and histology-specific differences in HPV prevalence and genotype distribution emphasize the need for tailored screening strategies, particularly for older women. These findings provide region-specific evidence to support the refinement of cervical cancer prevention and control strategies in China, particularly in contexts with similar demographic and epidemiological characteristics to the study population.