Evaluation and clinical application of a bracketing calibration-based isotope dilution liquid chromatography-tandem mass spectrometry candidate reference measurement procedure for serum theophylline.

A candidate reference measurement procedure (RMP) for serum theophylline via isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. With a single-step precipitation pretreatment and a 6-min gradient elution, the method achieved baseline separation of theophylline and its analogs on a C18-packed column. A bracketing calibration method was used to ensure repeatable signal intensity and high measurement precision. The intra-assay and inter-assay imprecisions were 1.06%, 0.84%, 0.72% and 0.47%, 0.41%, 0.25% at concentrations of 4.22 µg/mL (23.40 µmol/L), 8.45 µg/mL (46.90 µmol/L), and 15.21 µg/mL (84.43 µmol/L), respectively. Recoveries ranged from 99.35 to 102.34%. The limit of detection (LoD) was 2 ng/mL, and the lowest limit of quantification (LLoQ) was 5 ng/mL. The linearity range extended from 0.47 to 60 µg/mL (2.61-333.04 µmol/L). No ion suppression and carry-over (< 0.68%) were observed. The relative bias for this candidate RMP that participated in 2023 External Quality Control for Reference Laboratories (RELA) conducted by the International Federation of Clinical Chemistry (IFCC) was within a range of 0.17 to 0.93%. Furthermore, two clinical immunoassay systems were compared with this candidate RMP, demonstrating good correlations. The results of the Trueness Verification Plan indicate significant differences among routine systems, highlighting the need for standardization efforts. The developed candidate RMP for serum theophylline serves as a precise reference baseline for standardizing clinical systems and assigning values to reference materials.

Spatial-temporal variation and source analysis of heavy metals in different land use types in Beilun District (2015 and 2022).

The soil environment plays an important role in urban ecosystems. To study the heavy metal contamination of soil in Beilun District, Ningbo, we collected soil samples from 60 points in urban and peri-urban areas of Beilun District and analyzed the spatiotemporal variation and sources of heavy metal pollution in various land-use types. The results shown that the heavy metal contents in 2015 and 2022 were higher than the background soil values of Ningbo city, and there was an accumulation of heavy metals over these 7 years. The contents of heavy metals in green belts and woodland in 2022 were higher than those in 2015, while there was no significant change in agricultural land. The heavy metal contents in both years were mainly in the order green belts > agricultural land > woodland. The spatiotemporal distribution of heavy metal content showed that heavy metal pollution in Beilun District was concentrated in five industrial areas, and there was a trend toward the disappearance of highly polluted points. But the single-factor pollution index, pollution load index (PLI), and geoaccumulation index (Igeo) indicated that there was no significant heavy metal pollution in Beilun District, and individual elements at specific points showed slight pollution. The source analysis results showed that the main source of Hg is chemical, As is mainly derived from agricultural, Cr, Ni and Cu are mainly derived from natural, the main sources of Zn and Cd are electroplating and machinery activities, and the main source of Pb is traffic. These results specify a reference for future investigation on urban soil heavy metals, and the source apportionment results provide a scientific foundation for subsequent soil heavy metal pollution treatment.

Combination of Capture-SELEX and Post-SELEX for procymidone-specific aptamer selection and broad-specificity aptamer discovery, and development of aptamer-based lateral flow assay.

BACKGROUND: Due to its wide application, procymidone has become one of the pesticides with high detection rates in supervision and sampling. Therefore, it is necessary to establish a rapid and efficient method for the detection of procymidone. However, an important bottleneck restricting the development of rapid detection methods of procymidone is that its specific recognition elements are rarely reported. In this work, Capture-SELEX and post-SELEX were used in aptamer screening, and the obtained aptamers were used to construct an aptamer-based lateral flow assay (LFA). RESULTS: Firstly, a specific aptamer Seq15 was obtained for procymidone by Capture-SELEX, and its dissociation constant (Kd) was 24.22 nM. Secondly, post-SELEX was used to analyze and modify Seq15 to improve its performance, and the Kd of the truncated sequence Seq15-2 was 21.28 nM. In addition to this, the broad-specificity aptamer Seq17-1 was obtained via post-SELEX. Seq17-1 could broadly recognize dicarboximide fungicides (procymidone, iprodione, chlozolinate, dimethachlon and vinclozolin) and their metabolic derivative (3,5-dichloroaniline). Finally, the specific aptamer-based LFA of procymidone was constructed, and the limit of detection (LOD) was 0.79 ng/mL. Meanwhile, the LODs of dicarboximide fungicides and their metabolic derivative were 0.62, 0.64, 0.71, 0.69, 0.64 and 0.66 ng/mL, respectively. The above LFAs were highly specific and stable, and had been successfully used for the detection of vegetable samples. SIGNIFICANCE: Under the combination of Capture-SELEX and Post-SELEX, this study not only provides specific recognition elements for rapid detection of procymidone, but also provides new ideas for the discovery of broad-specificity aptamers. Combining broad-specificity primary detection and single-specificity quantification, a composite aptamer-based LFA detection platform has been developed, which significantly improves detection efficiency.

A personalized prognostic model for long-term survival in patients with intrahepatic cholangiocarcinoma: a retrospective cohort study.

Purpose: This study aimed to determine the optimal cutoff points for age and tumor size of patients with intrahepatic cholangiocarcinoma (ICC) and to establish and verify a predictive nomogram of overall survival at 1, 3, and 5 years. Methods: From the SEER (Surveillance, Epidemiology, and End Results) database, 1,325 ICC patients were selected and randomly divided into training and testing cohorts at a 7:3 ratio. Using the X-tile software, age and tumor size were classified into 3 subgroups: ≤61, 62-74, and ≥75 years and ≤35, 36-55, and ≥56 mm. Subsequently, univariate and multivariate Cox regression analyses were performed using the R software in the training cohort to determine independent risk factors, compile the prediction nomogram, and verify it with the testing cohort findings. Results: The C-indexes of the new prediction nomograms in the training and testing cohorts were 0.738 (95% confidence interval [CI], 0.718-0.758) and 0.750 (95% CI, 0.72-0.78), respectively. Furthermore, the areas under the 1-, 3-, and 5-year receiver operating characteristic (ROC) curves based on the nomogram were 0.792, 0.853, and 0.838, respectively, higher than the ROC based on the 7th and 8th editions of the American Joint Cancer Commission (AJCC) staging system. Conclusion: This study established and verified a prognostic nomogram that improved the accuracy of the 1-, 3-, and 5-year survival predictions for ICC patients, compared with that based on the 7th and 8th editions of the AJCC staging system, and can help clinicians make personalized survival predictions.

Multifunctional nanoenzyme lateral flow immunoassay strip for rapid and ultrasensitive detection of carbofuran in vegetables.

A lateral flow immunoassay strip (LFIAS) is one of the most frequently rapid test technologies for carbofuran (CAR). Nevertheless, the LFIAS has a poor quantitative capability and low sensitivity. And, it also requires often complex sample handling steps, making testing time longer. In this study, Fe3O4 nanoparticles were successively modified with MIL-100(Fe)-based metal-organic framework (MOF) and chloroplatinic acid hexahydrate to obtain a core-shell complex of Fe3O4-MOF-Pt. The complex had a peroxidase-mimicking activity catalytic function that enabled signal amplification and sensitivity enhancement. Upon coupling with carbofuran monoclonal antibody (CAR-mAb), the magnetic separation properties of the probe enabled target-specific enrichment. The LFIAS based on Fe3O4-MOF-Pt nanocomposites could detect CAR in the range of 0.25-50 ng mL-1 with a limit of detection (LOD) of 0.15 ng mL-1, enabling colorimetric and catalytic analysis. In addition, the method showed high specificity and stability for detecting CAR in various vegetables, and recovery rates of the spiked samples were 91.40%-102.40%. In conclusion, this study provided one-stop detection of "target enrichment-visual inspection". While lowering the LOD, it reduced the detection time and improved the detection efficiency. The multifunctional Fe3O4-MOF-Pt nanocomposite provides an idea for the construction of novel multifunctional probes to improve the detection performance of conventional LFIAS.

Ilizarov technique for treating elbow stiffness caused by myositis ossificans: A case report.

BACKGROUND: Myositis ossificans (MO) is a rare disease involving the formation of bone outside the musculoskeletal system. While surgical intervention is the main treatment approach, preventing recurrence and standardized rehabilitation are also crucial. Here, we present a surgical strategy to prevent the recurrence of MO. CASE SUMMARY: A 28-year-old female patient was admitted for the first time for a comminuted fracture of the left olecranon. However, incorrect postoperative rehabilitation resulted in the development of elbow joint stiffness with ectopic ossification, causing a loss of normal range of motion. The patient was diagnosed with MO based on physical examination, X-ray findings, and clinical presentation. We devised a surgical strategy to remove MO, followed by fixation with an Ilizarov frame, and implemented a scientifically reasonable rehabilitation plan. The surgery lasted for 3 h with an estimated blood loss of 45 mL. A drainage tube was placed after surgery, and fluid was aspirated through ultrasound-guided puncture. The patient experienced a significant reduction in joint stiffness after surgery. In the final follow-up at 9 mouths, there was evident improvement in the range of motion of the elbow joint, and no other symptoms were reported. CONCLUSION: The Ilizarov frame is an advantageous surgical technique for facilitating rehabilitation after MO removal. It offers benefits such as passive recovery, individualized treatment, and prompt recovery.

Direct Construction of 1,4-Dihydropyridazines and Pyrazoles via Annulation of Alkyl 2-Aroyl-1-chlorocyclopropanecarboxylates.

An efficient and chemodivergent synthesis of highly functionalized 1,4-dihydropyridazines and pyrazoles has been accomplished via base-promoted annulation between hydrazones and alkyl 2-aroyl-1-chlorocyclopropanecarboxylates, respectively. This transition-metal-free domino reaction proceeded rapidly under mild basic conditions, affording potentially bioactive 1,4-dihydropyridazine and pyrazole derivatives in moderate yields. The conversion of 1,4-dihydropyridazine to pyrazole was confirmed by adjusting the quantity of the base.

Establishment and clinical application of a candidate reference measurement procedure for quantification of urinary vanillylmandelic acid and homovanillic acid using ID-LC-MS/MS method.

Neuroblastoma (NB), an embryonic tumor of the autonomous nervous system, poses a significant threat to the health and lives of children. Accurate measurement of vanillylmandelic acid (VMA) and homovanillic acid (HVA) in human urine is crucial for screening and diagnosis of NB. Although various laboratories have developed liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect VMA and HVA, the comparability between the results obtained from different laboratories and methods was poor. The absence of reference method for VMA and HVA hinders the standardization of their measurements. Therefore, a candidate reference measurement procedure (cRMP) based on isotope dilution LC-MS/MS (ID-LC-MS/MS) for the detection of VMA and HVA in human urine was established. Urine samples were spiked with VMA-d3 and HVA-d5 as internal standards and extracted using a protein precipitation method. The cRMP exhibited desirable precision with the total imprecision below 5 %. The accuracy of this cRMP was demonstrated by the high analytical recovery (98.64 % - 102.22 % and 98.41 % - 100.97 % for VMA and HVA, respectively), and comparability between different reference systems. The limit of detection for HVA and VMA were 15.625 ng/mL and 3.906 ng/mL, respectively; the quantification limits were 62.5 ng/mL and 7.813 ng/mL, respectively, which can meet the clinical detection requirements. The linear range was from 78.125 ng/mL to 20 µg/mL. Specificity evaluations showed no corresponding interference from structurally similar analogs. In conclusion, we have established a cRMP based on ID-LC-MS/MS for the measurement of VMA and HVA in urine samples, demonstrating well-defined method performance including accuracy, precision, and specificity. This newly established cRMP is suitable for routine assay standardization and evaluation of clinical samples. Furthermore, this method has the potential to significantly enhance the diagnostic accuracy for neuroblastoma.

Three-dimensional ranging system based on Fresnel incoherent correlation holography.

We proposed a three-dimensional (3D) ranging system based on Fresnel incoherent correlation holography (FINCH). Distinct from the displacement measurement based on coherent digital holography (DH), our system simultaneously achieves a 3D range measurement using incoherent illumination. The observation range is obtained by the holographic reconstruction, while the in-plane range is determined using the two-dimensional digital imaging correlation (2D-DIC) technique. Experimental results on the resolution target demonstrate precise 3D ranging determination and improved measurement accuracy.

Synthesis of Polyethylene Terephthalate (PET) with High Crystallization and Mechanical Properties via Functionalized Graphene Oxide as Nucleation Agent.

In this work, a novel functionalized graphene oxide nucleating agent (GITP) was successfully synthesized using a silane coupling agent (IPTES), and polymer block (ITP) to efficiently improve the crystallization and mechanical performance of PET. To comprehensively investigate the effect of functionalized GO on PET properties, PET/GITP nanocomposites were prepared by introducing GITP into the PET matrix using the melt blending method. The results indicate that PET/GITP exhibits better thermal stability and crystallization properties compared with pure PET, increasing the melting temperature from 244.1 °C to 257.1 °C as well as reducing its crystallization half-time from 595 s to 201 s. Moreover, the crystallization temperature of PET/GITP nanocomposites was increased from 185.1 °C to 207.5 °C and the tensile strength was increased from 50.69 MPa to 66.8 MPa. This study provides an effective strategy for functionalized GO as a nucleating agent with which to improve the crystalline and mechanical properties of PET polyester.

Controlled attenuation parameter (CAP): the clinical value based on MRI-PDFF in children with obesity.

OBJECTIVES: Controlled attenuation parameter (CAP) is a noninvasive and quantitative method to evaluate hepatic steatosis, which is not well evaluated in children. The aim of this study was to examine the diagnostic value of CAP for hepatic steatosis in children with obesity based on MR proton density fat fraction (PDFF). METHODS: About 108 pediatric patients with nonalcoholic fatty liver disease (NAFLD) who were assessed for PDFF, CAP, and other laboratory results were enrolled. In this study, pediatric patients were separated for the obese group (n=80) and the severe obese group (n=28). Hepatic steatosis grades (0-3) were classified according to PDFF using cutoff values of 6.4 , 17.4, and 22.1 %. RESULTS: There are significant differences in CAP between the obese and severe obese groups (p<0.05). CAP showed a good correlation with PDFF in pediatric patients with NAFLD for diagnosing hepatic steatosis using a cutoff value of 265 dB/m (p<0.001). Meanwhile, ALT significantly outperforms CAP in receiver-operating curve (ROC) analysis for diagnosing hepatic steatosis grades. The diagnostic accuracy of CAP for steatosis is 77.8 %, and the diagnostic accuracy of ALT for steatosis is 83.3 %. CONCLUSIONS: While CAP holds promise as a diagnostic tool for pediatric NAFLD, its diagnostic performance warrants some caution. The potential of CAP is evident; however, ALT emerges as a simpler and more accurate measure for detecting hepatic steatosis in children. Further research is essential to determine the optimal role of CAP in pediatric NAFLD diagnosis and management.

Mortality outcomes in diabetic metabolic dysfunction-associated fatty liver disease: non-obese versus obese individuals.

The difference in the survival of obese patients and normal-weight/lean patients with diabetic MAFLD remains unclear. Therefore, we aimed to describe the long-term survival of individuals with diabetic MAFLD and overweight/obesity (OT2M), diabetic MAFLD with lean/normal weight (LT2M), MAFLD with overweight/obesity and without T2DM (OM), and MAFLD with lean/normal weight and without T2DM (LM). Using the NHANESIII database, participants with MAFLD were divided into four groups. Hazard ratios (HRs) and 95% confidence intervals (CIs) for all-cause, cardiovascular disease (CVD)-related, and cancer-related mortalities for different MAFLD subtypes were evaluated using Cox proportional hazards models. Of the 3539 participants, 1618 participants (42.61%) died during a mean follow-up period of 274.41 ± 2.35 months. LT2M and OT2M had higher risks of all-cause mortality (adjusted HR, 2.14; 95% CI 1.82-2.51; p < 0.0001; adjusted HR, 2.24; 95% CI 1.32-3.81; p = 0.003) and CVD-related mortality (adjusted HR, 3.25; 95% CI 1.72-6.14; p < 0.0001; adjusted HR, 3.36; 95% CI 2.52-4.47; p < 0.0001) than did OM. All-cause and CVD mortality rates in LT2M and OT2M patients were higher than those in OM patients. Patients with concurrent T2DM and MAFLD should be screened, regardless of the presence of obesity.

Distinguishable short-term effects of tea and water drinking on human saliva redox.

Food consumption can alter the biochemistry and redox status of human saliva, and the serving temperature of food may also play a role. The study aimed to explore the immediate (3 min) and delayed (30 min) effects of hot tea (57 ± 0.5 °C) ingestion and cold tea (8 ± 0.5 °C) ingestion on the salivary flow rate and salivary redox-relevant attributes. The saliva was collected from 20 healthy adults before, 3-min after and 30-min after the tea ingestion. The hot or cold deionised water at the same temperatures were used as control. The salivary flow rate and redox markers in hot tea (HBT), cold tea (CBT), hot water (HW) and cold water (CW) group were analysed and compared. The results demonstrated that neither the black tea nor the water altered the salivary flow rate; the black tea immediately increased the salivary thiol (SH) and malondialdehyde (MDA) content while reduced salivary uric acid (UA) significantly. The tea ingestion showed a tendency to elevate the ferric reducing antioxidant power (FRAP) in saliva, although not significantly. The water ingestion decreased the MDA content immediately and increased the UA level significantly. Cold water was found to induce a greater delayed increase in total salivary total protein (TPC) than the hot water. In conclusion, the black tea ingestion affects the redox attributes of human saliva acutely and significantly, while the temperature of drink makes the secondary contribution.

Whole-transcriptome sequencing reveals a melanin-related ceRNA regulatory network in the breast muscle of Xichuan black-bone chicken.

The economic losses incurred due to reduced muscle pigmentation highlight the crucial role of melanin-based coloration in the meat of black-bone chickens. Melanogenesis in the breast muscle of black-bone chickens is currently poorly understood in terms of molecular mechanisms. This study employed whole-transcriptome sequencing to analyze black and white breast muscle samples from black-bone chickens, leading to the identification of 367 differentially expressed (DE) mRNAs, 48 DElncRNAs, 104 DEcircRNAs, and 112 DEmiRNAs involved in melanin deposition. Based on these findings, a competitive endogenous RNA (ceRNA) network was developed to better understand the complex mechanisms of melanin deposition. Furthermore, our analysis revealed key DEmRNAs (TYR, DCT, EDNRB, MLPH and OCA2) regulated by DEmiRNAs (gga-miR-140-5p, gga-miR-1682, gga-miR-3529, gga-miR-499-3p, novel-m0012-3p, gga-miR-200b-5p, gga-miR-203a, gga-miR-6651-5p, gga-miR-7455-3p, gga-miR-31-5p, miR-140-x, miR-455-x, novel-m0065-3p, gga-miR-29b-1-5p, miR-455-y, novel-m0085-3p, and gga-miR-196-1-3p). These DEmiRNAs competitively interacted with DElncRNAs including MSTRG.2609.2, MSTRG.4185.1, LOC112530666, LOC112533366, LOC771030, LOC107054724, LOC121107411, LOC100859072, LOC101750037, LOC121108550, LOC121109224, LOC121110876, and LOC101749016, as well as DEcircRNAs, such as novel_circ_000158, novel_circ_000623, novel_001518, and novel_circ_003596. The findings from this study provide insight into the mechanisms that regulate lncRNA, circRNA, miRNA, and mRNA expression in chicken melanin deposition.

Edge enhancement in three-dimensional vortex imaging based on FINCH by Bessel-like spiral phase modulation.

Edge enhancement, as an important part of image processing, has played an essential role in amplitude-contrast and phase-contrast object imaging. The edge enhancement of three-dimensional (3D) vortex imaging has been successfully implemented by Fresnel incoherent correlation holography (FINCH), but the background noise and image contrast effects are still not satisfactory. To solve these issues, the edge enhancement of FINCH by employing Bessel-like spiral phase modulation is proposed and demonstrated. Compared with the conventional spiral phase modulated FINCH, the proposed technique can achieve high-quality edge enhancement 3D vortex imaging with lower background noise, higher contrast and resolution. The significantly improved imaging quality is mainly attributed to the effective sidelobes' suppression in the generated optical vortices with the Bessel-like modulation technique. Experimental results of the small circular aperture, resolution target, and the Drosophila melanogaster verify its excellent imaging performance. Moreover, we also proposed a new method for selective edge enhancement of 3D vortex imaging by breaking the symmetry of the spiral phase in the algorithmic model of isotropic edge enhancement. The reconstructed images of the circular aperture show that the proposed method is able to enhance the edges of the given objects selectively in any desired direction.

[Multi-modal physiological time-frequency feature extraction network for accurate sleep stage classification].

Sleep stage classification is essential for clinical disease diagnosis and sleep quality assessment. Most of the existing methods for sleep stage classification are based on single-channel or single-modal signal, and extract features using a single-branch, deep convolutional network, which not only hinders the capture of the diversity features related to sleep and increase the computational cost, but also has a certain impact on the accuracy of sleep stage classification. To solve this problem, this paper proposes an end-to-end multi-modal physiological time-frequency feature extraction network (MTFF-Net) for accurate sleep stage classification. First, multi-modal physiological signal containing electroencephalogram (EEG), electrocardiogram (ECG), electrooculogram (EOG) and electromyogram (EMG) are converted into two-dimensional time-frequency images containing time-frequency features by using short time Fourier transform (STFT). Then, the time-frequency feature extraction network combining multi-scale EEG compact convolution network (Ms-EEGNet) and bidirectional gated recurrent units (Bi-GRU) network is used to obtain multi-scale spectral features related to sleep feature waveforms and time series features related to sleep stage transition. According to the American Academy of Sleep Medicine (AASM) EEG sleep stage classification criterion, the model achieved 84.3% accuracy in the five-classification task on the third subgroup of the Institute of Systems and Robotics of the University of Coimbra Sleep Dataset (ISRUC-S3), with 83.1% macro F1 score value and 79.8% Cohen's Kappa coefficient. The experimental results show that the proposed model achieves higher classification accuracy and promotes the application of deep learning algorithms in assisting clinical decision-making.

Tanshinone IIA modulates cancer cell morphology and movement via Rho GTPases-mediated actin cytoskeleton remodeling.

Actin filaments form unique structures with robust actin bundles and cytoskeletal networks affixed to the extracellular matrix and interact with neighboring cells, which are crucial structures for cancer cells to acquire a motile phenotype. This study aims to investigate a novel antitumor mechanism by which Tanshinone IIA (Tan IIA) modulates the morphology and migration of liver cancer cells via actin cytoskeleton regulation. 97H and Huh7 exhibited numerous tentacle-like protrusions that interacted with neighboring cells. Following treatment with Tan IIA, 97H and Huh7 showed a complete absence of cytoplasmic protrusion and adherens junctions, thereby effectively impeding their migration capability. The fluorescence staining of F-actin and microtubules indicated that these tentacle-like protrusions and cell-cell networks were actin-based structures that led to morphological changes after Tan IIA treatment by retracting and reorganizing beneath the membrane. Tan IIA can reverse the actin depolymerization and cell morphology alterations induced by latrunculin A. Tan IIA down-regulated actin and Rho GTPases expression significantly, as opposed to inducing Rho signaling activation. Preventing the activity of proteasomes and lysosomes had no discernible impact on the modifications in cellular structure and protein expression induced by Tan IIA. However, as demonstrated by the puromycin labeling technique, the newly synthesized proteins were significantly inhibited by Tan IIA. In conclusion, Tan IIA can induce dramatic actin cytoskeleton remodeling by inhibiting the protein synthesis of actin and Rho GTPases, resulting in the suppression of tumor growth and migration. Targeting the actin cytoskeleton of Tan IIA is a promising strategy for HCC treatment.