A novel analytical method was developed in this study by combining online solid phase extraction with ultra performance liquid chromatography-tandem mass spectrometry(Online SPE-UPLC-MS/MS)for simultaneous determination of 50 kinds of steroid hormones in surface water.Specifically,after high-speed centrifugation of 4 mL water samples,the supernatant was directly injected into an Oasis HLB online SPE column for enrichment and purification.Subsequently,the target compounds were transferred to the analytical column via valve switching for separation and analysis.The chromatographic separation was performed on a Thermo Acclaim RSLC C18 column(100 mm×2.1 mm,2.2 μm),using a mobile phase composed of 5 mmol/L ammonium fluoride aqueous solution and acetonitrile.Mass spectrometric detection was conducted in positive ion mode,utilizing multiple reaction monitoring(MRM)with quantification achieved by the internal standard method.The method validation demonstrated that the limits of detection(LOD)for the 50 kinds of steroid hormones ranged from 0.02 to 0.50 ng/L,while the limits of quantification(LOQ)were between 0.08 and 1.67 ng/L.The average recoveries in surface water samples at spiked concentrations of 5,20 and 200 ng/L were between 74.1％and 119％,with relative standard deviations(RSDs)of 0.2％to 9.9％.This method was applied to analyze 11 surface water samples collected from sites surrounding a pharmaceutical and chemical industrial park.A total of 44 kinds of steroid hormones were detected,with concentrations ranging from 0.11 to 88.6 ng/L,revealing the presence of hormone contamination in the environmental waters surrounding industrial areas.Compared with the traditional offline SPE methods,the proposed online SPE technique significantly reduced sample volume requirements and pretreatment time,while minimizing the loss of target compounds during the pretreatment process.Moreover,compared to reported online SPE techniques,this method achieved high-throughput analysis of multiple classes of steroid hormones,with lower detection limits and higher recoveries.Overall,this method provided rapid sample preparation,high sensitivity,and excellent stability,making it suitable for the direct analysis of trace steroid hormones in surface water.

Protein tyrosine phosphatase nonreceptor type 2 (PTPN2) is a promising target for sensitizing solid tumors to immune checkpoint blockades. However, the highly polar active sites of PTPN2 hinder drug discovery efforts. Leveraging small interfering RNA (siRNA) technology, we developed a novel glutathione-responsive nano-platform HPssPT (HA/PEIss@siPtpn2) to silence PTPN2 and enhance immunotherapy efficacy in hepatocellular carcinoma (HCC). HPssPT showed potent transfection and favorable safety profiles. PTPN2 deficiency induced by HPssPT amplified the interferon γ signaling in HCC cells by increasing the phosphorylation of Janus-activated kinase 1 and signal transducer and activator of transcription 1, resulting in enhanced antigen presentation and T cell activation. The nano-platform was also able to promote the M1-like polarization of macrophages in vitro. The unique tropism of HPssPT towards tumor-associated macrophages, facilitated by hyaluronic acid coating and CD44 receptor targeting, allowed for simultaneous reprogramming of both tumor cells and tumor-associated macrophages, thereby synergistically reshaping tumor microenvironment to an immunostimulatory state. In HCC, colorectal cancer, and melanoma animal models, HPssPT monotherapy provoked robust antitumor immunity, thereby sensitizing tumors to PD-1 blockade, which provided new inspiration for siRNA-based drug discovery and tumor immunotherapy.

OBJECTIVE: To identify the key features of facial and tongue images associated with anemia in female populations, establish anemia risk-screening models, and evaluate their performance. METHODS: A total of 533 female participants (anemic and healthy) were recruited from Shuguang Hospital. Facial and tongue images were collected using the TFDA-1 tongue and face diagnosis instrument. Color and texture features from various parts of facial and tongue images were extracted using Face Diagnosis Analysis System (FDAS) and Tongue Diagnosis Analysis System version 2.0 (TDAS v2.0). Least Absolute Shrinkage and Selection Operator (LASSO) regression was used for feature selection. Ten machine learning models and one deep learning model (ResNet50V2 + Conv1D) were developed and evaluated. RESULTS: Anemic women showed lower a-values, higher L- and b-values across all age groups. Texture features analysis showed that women aged 30-39 with anemia had higher angular second moment (ASM)and lower entropy (ENT) values in facial images, while those aged 40-49 had lower contrast (CON), ENT, and MEAN values in tongue images but higher ASM. Anemic women exhibited age-related trends similar to healthy women, with decreasing L-values and increasing a-, b-, and ASM-values. LASSO identified 19 key features from 62. Among classifiers, the Artificial Neural Network (ANN) model achieved the best performance [area under the curve (AUC): 0.849, accuracy: 0.781]. The ResNet50V2 model achieved comparable results [AUC: 0.846, accuracy: 0.818]. CONCLUSION Differences in facial and tongue images suggest that color and texture features can serve as potential TCM phenotype and auxiliary diagnostic indicators for female anemia.
Humans ; Female ; Tongue/diagnostic imaging* ; Adult ; Anemia/diagnosis* ; Middle Aged ; Face/diagnostic imaging* ; Young Adult ; Machine Learning

The study aims to examine the effects and potential mechanisms of disulfiram (DSF) on cardiac hypertrophic injury, focusing on the role of transforming growth factor-β-activated kinase 1 (TAK1)-mediated pan-apoptosis (PANoptosis). H9C2 cardiomyocytes were treated with angiotensin II (Ang II, 1 µmol/L) to establish an in vitro model of myocardial hypertrophy. DSF (40 µmol/L) was used to treat cardiomyocyte hypertrophic injury models, either along or in combination with the TAK1 inhibitor, 5z-7-oxozeaenol (5z-7, 0.1 µmol/L). We assessed cell damage using propidium iodide (PI) staining, measured cell viability with CCK8 assay, quantified inflammatory factor levels in cell culture media via ELISA, detected TAK1 and RIPK1 binding rates using immunoprecipitation, and analyzed the protein expression levels of key proteins in the TAK1-mediated PANoptosis pathway using Western blot. In addition, the surface area of cardiomyocytes was measured with Phalloidin staining. The results showed that Ang II significantly reduced the cellular viability of H9C2 cardiomyocytes and the binding rate of TAK1 and RIPK1, significantly increased the surface area of H9C2 cardiomyocytes, PI staining positive rate, levels of inflammatory factors [interleukin-1β (IL-1β), IL-18, and tumor necrosis factor α (TNF-α)] in cell culture media and p-TAK1/TAK1 ratio, and significantly up-regulated key proteins in the PANoptosis pathway [pyroptosis-related proteins NLRP3, Caspase-1 (p20), and GSDMD-N (p30), apoptosis-related proteins Caspase-3 (p17), Caspase-7 (p20), and Caspase-8 (p18), as well as necroptosis-related proteins p-MLKL, RIPK1, and RIPK3]. DSF significantly reversed the above changes induced by Ang II. Both 5z-7 and exogenous IL-1β weakened these cardioprotective effects of DSF. These results suggest that DSF may alleviate cardiac hypertrophic injury by inhibiting TAK1-mediated PANoptosis.
Animals ; MAP Kinase Kinase Kinases/physiology* ; Rats ; Myocytes, Cardiac/pathology* ; Disulfiram/pharmacology* ; Cardiomegaly ; Apoptosis/drug effects* ; Cell Line ; Angiotensin II ; Necroptosis/drug effects* ; Interleukin-1beta/metabolism* ; Receptor-Interacting Protein Serine-Threonine Kinases/metabolism* ; Lactones ; Resorcinols ; Zearalenone/administration & dosage*

In recent years, an increasing number of emerging environmental pollutants have been identified, garnering widespread attention. Many of these pollutants are characterized by their environmental persistence and bioaccumulation, which pose significant threats to both the ecological environment and human health. However, the molecular mechanisms underlying their effects remain unclear, limiting our ability to assess their adverse impacts and develop effective protective measures. The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor traditionally known to be activated by dioxins and polycyclic aromatic hydrocarbons (PAHs) and is involved in the metabolism of exogenous chemicals. Recent research has shown that the AHR can be activated by a diverse range of exogenous and endogenous chemicals and participates in various biological processes. Studies have demonstrated that AHR mediates the toxic effects of emerging environmental pollutants such as perfluorooctane sulfonamide (PFOSA) and N-(1,3-dimethylbutyl)-N’-phenyl-p-phenylenediamine quinone (6PPDQ). This paper provided an overview of the AHR activation and the toxic effects induced by emerging environmental pollutants, with a focus on how the AHR activation interacts with multiple signaling pathways. The significance of these interactions in environmental risk assessment and toxicological research was also discussed. We aim to provide a scientific basis for environmental protection and risk assessment.

Objective:To evaluate the effect of umbilical cord milking (UCM) on neonatal outcomes after cesarean section.Methods:Chinese and English databases (including CNKI, Wanfang, China Biology Medicine Disc, VIP, Yiigle, PubMed, Embase, CINAHL, Web of Science, the Cochrane Library, and Google Scholar) and ClinicalTrials.gov were retrieved from the inception to July 2023. Randomized controlled trials regarding UCM in neonates from different races who were born by cesarean section were included. The outcomes were postnatal hemoglobin level, hematocrit value, peak serum bilirubin level, phototherapy, cord blood pH value, intraventricular hemorrhage, death, polycythemia, neonatal necrotizing enterocolitis, and Apgar score. The risk of bias among the included studies was confined to low or possible risk according to the Cochrane Risk of Bias Assessment Tool 2.0. RevMan5.3 was used for meta-analysis, and subgroup analysis was performed among neonates with different gestational ages. The certainty of evidence was evaluated using the grades of recommendations assessment, development, and evaluation (GRADE) framework.Results:A total of 11 articles involving 2 347 neonates (1 322 full-term and 1 025 preterm infants) were included. Meta-analysis results showed that: (1) Compared with the immediate cord clamping, UCM increased the hemoglobin level within 24 h and 48-72 h after birth ( MD=1.40, 95% CI: 1.11-1.70, Z=9.32; MD=0.86, 95% CI: 0.69-1.02, Z=10.02, both P<0.01), hematocrit value within 24 h and 48-72 h after birth ( MD=2.73, 95% CI: 0.18-5.29, Z=2.09, P=0.04; MD=3.57, 95% CI: 2.29-4.85, Z=5.46, P<0.01). However, no significant differences were found in the peak bilirubin level, phototherapy, cord blood pH, and Apgar score at 1 and 5 min (all P>0.05). (2) Compared with delayed cord clamping, UCM increased the hemoglobin level ( MD=0.83, 95% CI: 0.75-0.91, Z=20.11, P<0.01) and hematocrit value ( MD=2.34, 95% CI: 1.25-3.43, Z=4.20, P<0.01) within 24 h after birth, but not in the hematocrit value at 48-72 h after birth ( MD=－0.38, 95% CI:－2.27-1.52, Z=0.39, P=0.70) or the peak bilirubin level ( MD=－0.65, 95% CI:－2.16-1.04, Z=0.69, P=0.49). Sensitivity analysis showed that for full-term neonates born by cesarean section, the peak bilirubin level in the UCM group was significantly lower than that in the delayed cord clamping group ( MD=－1.30, 95% CI:－2.26-0.34, Z=2.66, P<0.01). Still, the incidence of phototherapy, intraventricular hemorrhage (grade Ⅰ-Ⅳ), death, polycythemia, neonatal necrotizing enterocolitis, and Apgar score at 1 min and 5 min showed no statistical differences (all P>0.05). Conclusions:UCM could increase the short-term postnatal hemoglobin and hematocrit levels in neonates born by cesarean section, which might prevent neonatal anemia in the short term without increasing the adverse neonatal outcomes. Little effects were observed on the peak bilirubin level, phototherapy, polycythemia, etc. More high-quality and large-sample randomized controlled trials are needed in the future.

Objective To study the ability of radial basis function neural network(RBFNN)with different implementations for electrical impedance tomography(EIT)under real brain shapes,to evaluate the advantages and disadvantages of different approaches,and to provide a reference for the selection of practical imaging methods.Methods COMSOL Multiphysics was used to establish a multilayer 2D model with real structure based on brain CT and an EIT simulation dataset.The effects of the exact RBFNN,the orthogonal least squares-based RBFNN(OLS RBFNN)and the K-Means-based BRFNN(K-Means RBFNN)on the image reconstruction result were explored with the dataset constructed.The root mean square error(RMSE)and image correlation coefficient(ICC)were adopted to evaluate the imaging results.Results EIT could be completed with all the three RBFNNs without noise,and the exact RBFNN had the best results with average ICC and RMSE of 0.784 and 0.467,respectively,in the test set.The OLS RBFNN had the best imaging results at a hidden node of 50,with an average ICC and RMSE of 0.788 and 0.462,respectively.The K-Means RBFNN achieved the best imaging results at noise levels of 30,40,50,60,70 and 80 dB with stable ICC and RMSE and high robustness.Conclusion All the three RBFNNs can be used for brain EIT image reconstruction with their own advantages and disadvantages,and the RBFNN has to be selected for EIT reconstruc-tion based on considerations on actual conditions.[Chinese Medical Equipment Journal,2024,45(10):1-6]

Objective To investigate the relationship between gene mutation characteristics and immunological markers in patients with small cell lung cancer.Methods From January 2019 to 2020,155 patients with small cell lung cancer were admitted.Gene mutations were detected in these patients using target gene capture and sequencing method.Moreover,the tumor mutational burden(TMB)and expression of PD-L1 in some patients were detected.Results A total of 326 gene mutations were detected in the patients,the most significant of which were TP53 and RB1.Among the 8 genes with mutation frequency higher than 15％,patients with KMT2D,KMT2C,FAT1 and NOTCH1 mutations had higher TMB than those without mutation,and the difference was statistically significant(P＜0.05).Conclusion The most common mutated genes in patients with small cell lung cancer are TP53 and RB1.KMT2D,KMT2C,FAT1 and NOTCH1 may be potential markers for the efficacy of immunotherapy in small cell lung cancer.

OBJECTIVE Aiming at the current vacancy of large language models(LLMs)in TCM evaluation,a TCM benchmark dataset is designed and constructed to comprehensively and objectively evaluate the mastery and reasoning performance of LLMs in TCM knowledge,providing scientific and reliable basis for optimizing the performance of LLMs in the field of TCM.METHODS This benchmark includes 29 506 questions across 13 subjects,with data collected from standardized TCM exams and textbooks.Three gen-eral-purpose models(GPT-3.5,ChatGLM3,Baichuan)and five Chinese medical models(PULSE,BenTsao,HuatuoGPT2,Bian-Que2,ShenNong)were evaluated with answer prediction and answer reasoning tasks.The evaluation results were quantitatively as-sessed using metrics including accuracy,F1 score,BLEU,and Rouge.RESULTS For the answer prediction task,Baichuan had the highest accuracy of 36.07%in single-choice questions,while ChatGLM3 achieved the highest accuracy of 18.96%and F1 score of 76.31%in multiple-choice questions.For the answer reasoning experiment,Baichuan scored highest on BLEU-1 with 24.71,while ChatGLM3 achieved the highest Rouge-1 score of 44.64.CONCLUSION In this study,general LLMs performed slightly better than Chinese medical LLMs.Meanwhile,all models'accuracy on choice questions remained below 60%,reflecting the significant challen-ges and room for improvement that LLMs still face in the field of TCM.

Knowledge about the neuronal dynamics and the projectome are both essential for understanding how the neuronal network functions in concert. However, it remains challenging to obtain the neural activity and the brain-wide projectome for the same neurons, especially for neurons in subcortical brain regions. Here, by combining in vivo microscopy and high-definition fluorescence micro-optical sectioning tomography, we have developed strategies for mapping the brain-wide projectome of functionally relevant neurons in the somatosensory cortex, the dorsal hippocampus, and the substantia nigra pars compacta. More importantly, we also developed a strategy to achieve acquiring the neural dynamic and brain-wide projectome of the molecularly defined neuronal subtype. The strategies developed in this study solved the essential problem of linking brain-wide projectome to neuronal dynamics for neurons in subcortical structures and provided valuable approaches for understanding how the brain is functionally organized via intricate connectivity patterns.
Animals ; Neurons/physiology* ; Mice ; Brain/physiology* ; Mice, Inbred C57BL ; Somatosensory Cortex/physiology* ; Neural Pathways/physiology* ; Hippocampus/physiology* ; Mice, Transgenic ; Male ; Brain Mapping ; Nerve Net/physiology* ; Substantia Nigra/physiology* ; Tomography, Optical/methods*