Dual-Mode Fluorescent/Intelligent Lateral Flow Immunoassay Based on Machine Learning Algorithm for Ultrasensitive Analysis of Chloroacetamide Herbicides.

Given the harmful effect of pesticide residues, it is essential to develop portable and accurate biosensors for the analysis of pesticides in agricultural products. In this paper, we demonstrated a dual-mode fluorescent/intelligent (DM-f/DM-i) lateral flow immunoassay (LFIA) for chloroacetamide herbicides, which utilized horseradish peroxidase-IgG conjugated time-resolved fluorescent nanoparticle probes as both a signal label and amplification tool. With the newly developed LFIA in the DM-f mode, the limits of detection (LODs) were 0.08 ng/mL of acetochlor, 0.29 ng/mL of metolachlor, 0.51 ng/mL of Propisochlor, and 0.13 ng/mL of their mixture. In the DM-i mode, machine learning (ML) algorithms were used for image segmentation, feature extraction, and correlation analysis to obtain multivariate fitted equations, which had high reliability in the regression model with R2 of 0.95 in the range of 2 × 102-2 × 105 pg/mL. Importantly, the practical applicability was successfully validated by determining chloroacetamide herbicides in the corn sample with good recovery rates (85.4 to 109.3%) that correlate well with the regression model. The newly developed dual-mode LFIA with reduced detection time (12 min) holds great potential for pesticide monitoring in equipment-limited environments using a portable test strip reader and laboratory conditions using ML algorithms.

Microstructure-Driven Self-Transport and Convection of Water on Membrane Surface for Ultra-Fast, Highly Sensitive, Low-Cost Lateral-Flow Assays.

Lateral-flow assay (LFA) is one of the most commonly used detection technologies, in which the chromatographic membranes are currently used as the lateral-flow membrane (e.g., nitrocellulose membrane, NC Mem). However, several disadvantages of existing chromatographic membranes limit the performance of LFA, including relatively low flow velocity of sample solution and relatively more residuals of sample on membrane, which increase detection time and detection noise. Herein, a surface structure membrane (SS Mem) is proposed, which enables fast self-transport of water with a convection manner and realizes low residuals of sample on membrane surface after the flow. On SS Mem, the flow velocity of water is 7.1-fold higher, and the residuals of sample are decreased by 60-67%, comparing those in NC Mem. SS Mem is used as lateral-flow membrane to prepare lateral-flow strips of nanogold LFA and fluorescence LFA for rapid detection of SARS CoV-2 nucleocapsid protein. These LFAs require 210 s per detection, with limits of detection of 3.98 pg mL-1 and 53.3 fg mL-1, sensitivity of 96.5%, and specificity of 90%. The results suggest that SS Mem enables ultrafast, highly sensitive lateral-flow immunoassays and shows great potential as a new type of lateral-flow membrane to broaden the application of LFA.

Are children with IgA nephropathy different from adult patients?

BACKGROUND: Previously, several studies have indicated that pediatric IgA nephropathy (IgAN) might be different from adult IgAN, and treatment strategies might be also different between pediatric IgAN and adult IgAN. METHODS: We analyzed two prospective cohorts established by pediatric and adult nephrologists, respectively. A comprehensive analysis was performed investigating the difference in clinical and pathological characteristics, treatment, and prognosis between children and adults with IgAN. RESULTS: A total of 1015 children and 1911 adults with IgAN were eligible for analysis. More frequent gross hematuria (88% vs. 20%, p < 0.0001) and higher proteinuria (1.8 vs. 1.3 g/d, p < 0.0001) were seen in children compared to adults. In comparison, the estimated glomerular filtration rate (eGFR) was lower in adults (80.4 vs. 163 ml/min/1.73 m2, p < 0.0001). Hypertension was more prevalent in adult patients. Pathologically, a higher proportion of M1 was revealed (62% vs. 39%, p < 0.0001) in children than in adults. S1 (62% vs. 28%, p < 0.0001) and T1-2 (34% vs. 8%, p < 0.0001) were more frequent in adults. Adjusted by proteinuria, eGFR, and hypertension, children were more likely to be treated with glucocorticoids than adults (87% vs. 45%, p < 0.0001). After propensity score matching, in IgAN with proteinuria > 1 g/d, children treated with steroids were 1.87 (95% CI 1.16-3.02, p = 0.01) times more likely to reach complete remission of proteinuria compared with adults treated with steroids. CONCLUSIONS: Children present significantly differently from adults with IgAN in clinical and pathological manifestations and disease progression. Steroid response might be better in children.

A low-voltage alternant direct current electroporation chip for ultrafast releasing the genome DNA of Helicobacter pylori bacterium.

Nucleic acid detection, as an important molecular diagnostic method, is widely used in bacterial identification, disease diagnosis. For detecting the nucleic acid of bacteria, the prerequisite is to release nucleic acids inside the bacteria. The common means to release nucleic acids is the chemical method, which involves complex processes, is time-consuming, and remains chemical inhibitors. Compared with chemical methods, electroporation as a physical method has the advantages of easy operation, short-time consumption, and chemical reagents free. However, the current works using electroporation often necessitates high-frequency or high-voltage conditions, entailing bulky power devices. Herein, we propose a low-voltage alternant direct current (LADC) electroporation chip and the corresponding miniature device for ultrafast releasing the genome DNA from Helicobacter pylori (H. pylori) for detection. We connected a micrometer-interdigital electrode in the chip with a 20 V portable battery to make the miniature device. Using this low-voltage device, our chip released genome DNA of H. pylori within only 5 ms, achieving a cell lysis rate of 99.5%. We further combined this chip with a colorimetric loop-mediated isothermal amplification assay to visually detect H. pylori within ~ 25 min at 10 CFU/µL. We detected 11 clinical samples using the chip, and the detection results were consistent with those of the clinical standard. The results indicate that the LADC electroporation chip is useful for ultrafast release of genome DNA from bacteria and is expected to promote the development of nucleic acid detection in POCT and other scenarios.

Intelligent, Flexible Artificial Throats with Sound Emitting, Detecting, and Recognizing Abilities.

In recent years, there has been a notable rise in the number of patients afflicted with laryngeal diseases, including cancer, trauma, and other ailments leading to voice loss. Currently, the market is witnessing a pressing demand for medical and healthcare products designed to assist individuals with voice defects, prompting the invention of the artificial throat (AT). This user-friendly device eliminates the need for complex procedures like phonation reconstruction surgery. Therefore, in this review, we will initially give a careful introduction to the intelligent AT, which can act not only as a sound sensor but also as a thin-film sound emitter. Then, the sensing principle to detect sound will be discussed carefully, including capacitive, piezoelectric, electromagnetic, and piezoresistive components employed in the realm of sound sensing. Following this, the development of thermoacoustic theory and different materials made of sound emitters will also be analyzed. After that, various algorithms utilized by the intelligent AT for speech pattern recognition will be reviewed, including some classical algorithms and neural network algorithms. Finally, the outlook, challenge, and conclusion of the intelligent AT will be stated. The intelligent AT presents clear advantages for patients with voice impairments, demonstrating significant social values.

Focus on Mitochondrial Respiratory Chain: Potential Therapeutic Target for Chronic Renal Failure.

The function of the respiratory chain is closely associated with kidney function, and the dysfunction of the respiratory chain is a primary pathophysiological change in chronic kidney failure. The incidence of chronic kidney failure caused by defects in respiratory-chain-related genes has frequently been overlooked. Correcting abnormal metabolic reprogramming, rescuing the "toxic respiratory chain", and targeting the clearance of mitochondrial reactive oxygen species are potential therapies for treating chronic kidney failure. These treatments have shown promising results in slowing fibrosis and inflammation progression and improving kidney function in various animal models of chronic kidney failure and patients with chronic kidney disease (CKD). The mitochondrial respiratory chain is a key target worthy of attention in the treatment of chronic kidney failure. This review integrated research related to the mitochondrial respiratory chain and chronic kidney failure, primarily elucidating the pathological status of the mitochondrial respiratory chain in chronic kidney failure and potential therapeutic drugs. It provided new ideas for the treatment of kidney failure and promoted the development of drugs targeting the mitochondrial respiratory chain.

The Reciprocal Relations between Parental Psychological Control and Social Anxiety and the Mediating Role of Self-Concept Clarity Among Chinese Early Adolescents.

Although parental psychological control has been well-documented as a significant predictor of social anxiety among adolescents, few studies examine how changes in parental psychological control and adolescent social anxiety are reciprocally related at the within-person level, especially in Chinese culture. This longitudinal study examined reciprocal relations between parental psychological control and social anxiety, and the potential mediating role of self-concept clarity, by disentangling between- and within-person effects. A total of 4731 students (44.9% girls; Mage = 10.91 years, SD = 0.72) participated in a four-wave longitudinal study with 6-month intervals. Results from random intercept cross-lagged panel modeling indicated that parental psychological control directly predicted social anxiety, and vice versa. Parental psychological control indirectly predicted social anxiety via self-concept clarity, and social anxiety also indirectly predicted parental psychological control via self-concept clarity. These findings reveal a vicious cycle of mutual influence between parental psychological control and adolescent social anxiety in Chinese youth, and highlight the crucial role of self-concept clarity in the interplay between parenting and adolescent social functioning.

Maternal Childhood Maltreatment and Adolescent Internalizing Problems: The Mediating Role of Maternal Depressive Symptoms and Offspring Childhood Maltreatment.

Research has demonstrated the predictive effect of maternal childhood maltreatment on adolescent internalizing problems. However, few studies have explored the mediating mechanisms of how mothers' experiences of childhood maltreatment are transmitted to their offspring's internalizing problems over time. The present multi-informant study investigated the potential mediating effects of maternal depressive symptoms and offspring's childhood maltreatment experiences on the relation between maternal childhood maltreatment and adolescent internalizing problems. A total of 823 Chinese youth (43.4% girls; Mage = 10.26 years, SD = 0.94) and their mothers participated in a two-wave longitudinal study with one-year intervals. Mothers reported their experiences of childhood maltreatment and depressive symptoms, while youth reported their childhood maltreatment experiences and internalizing problems. Findings of path analysis indicated that maternal emotional abuse at T1 could significantly predict adolescent internalizing problems at T2, after controlling for a baseline of adolescent internalizing problems. Maternal emotional abuse, emotional neglect, and physical neglect at T1 can influence adolescent internalizing problems at T2 through maternal depressive symptoms at T1 to adolescent internalizing problems at T1. Maternal emotional abuse at T1 displayed statistically significant indirect effects on adolescent internalizing problems at T2 successively through the pathway from adolescent emotional abuse at T1 to adolescent internalizing problems at T1. The findings supported the cycle of maltreatment hypothesis. The present study highlights the intergenerational link between maternal childhood maltreatment and adolescent internalizing problems, as well as reveals the mediating mechanisms in this relation.

Social Anxiety and Peer Victimization and Aggression: Examining Reciprocal Trait-State Effects among Early Adolescents.

As peer relationships become paramount during early adolescence, there's a normative rise in social anxiety, coinciding with a peak in peer victimization and aggression. Although previous studies have suggested reciprocal associations between changes in social anxiety and adolescent peer victimization and aggression, the mechanics of these associations at the personal trait and time-varying state levels remains unclear. This study examined the longitudinal relations between social anxiety and adolescent peer victimization and aggression by disentangling between-person trait differences from within-person state processes. A total of 4731 Chinese early adolescents (44.9% girls; M age = 10.91 years, SD = 0.72) participated in a four-wave longitudinal study with 6-month intervals. Random-intercept cross-lagged panel model (RI-CLPM) was applied. The results revealed higher levels of social anxiety are associated with more peer victimization and aggression at the between-person trait level. At the within-person state level, adolescent social anxiety, and adolescent physical victimization and physical aggression, reciprocally predicted each other. Relational victimization significantly predicted an increase of social anxiety, but not vice versa. Social anxiety positively predicted relational aggression over time, whereas the effect of relational aggression on social anxiety was only observed at the initial stage of early adolescence. These findings highlight that various types of victimization and aggression might exhibit unique reciprocal associations with social anxiety. Distinguishing between the within-person state and between-person trait effects is crucial in research that informs the co-development of adolescent peer victimization, aggression, and social anxiety.

SR-Unet: A Super-Resolution Algorithm for Ion Trap Mass Spectrometers Based on the Deep Neural Network.

The mass spectrometer is an important tool for modern chemical analysis and detection. Especially, the emergence of miniature mass spectrometers has provided new tools for field analysis and detection. The resolution of a mass spectrometer reflects the ability of the instrument to discriminate between adjacent mass-to-charge ratio ions, and the higher the resolution, the better the discrimination of complex mixtures. Quadrupole ion traps are generally considered as a low-resolution mass spectrometry method, but they have gained wide attention and development in recent years because of their suitability for miniaturization and high qualitative capability. For an ion trap mass spectrometer, the mass sensitivity and resolution can be mutually constrained and need to be balanced by setting an appropriate scanning speed. In this study, a super-resolution U-net algorithm (SR-Unet) is proposed for ion trap mass spectrometry, which can estimate the possible ions from the overlapping ion peaks of low-resolution spectra and improve the equivalent resolution while ensuring sufficient sensitivity and analysis speed of the instrument. By determining the mass spectra of a linear ion trap mass spectrometer (LTQ XL) in Turbo and Normal scan modes, the same unit mass resolution as that at a scan speed of 16,667 Da/s was successfully obtained at 125,000 Da/s. Also, the experiments demonstrated that the algorithm is capable of the mass-to-charge ratio and instrument migration. SR-Unet can be migrated and applied to a miniature mass spectrometer for cruise detection of volatile organic compounds (VOCs), and the identification of VOC species in Photochemical Assessment Monitoring Stations (PAMS) was improved from 31 to 50 species with the same monitoring and analysis speed requirement. Further, super-unit mass resolution peptide detection was achieved on a miniature mass spectrometer with the help of the SR-Unet algorithm, which reduced the full width at half-maxima (FWHM) of bradykinin divalent ions (m/z 531) from 0.35 to 0.15 Da at a scan speed of 375 Da/s and improved the equivalent resolution to 3540. The proposed method provides a new idea to enhance the field mixture detection capability of miniature ion trap mass spectrometers.

PddCas: A Polydisperse Droplet Digital CRISPR/Cas-Based Assay for the Rapid and Ultrasensitive Amplification-Free Detection of Viral DNA/RNA.

Clustered regularly interspaced short palindromic repeats (CRISPR)-based assays have been an emerging diagnostic technology for pathogen diagnosis. In this work, we developed a polydisperse droplet digital CRISPR-Cas-based assay (PddCas) for the rapid and ultrasensitive amplification-free detection of viral DNA/RNA with minimum instruments. LbaCas12a and LbuCas13a were used for the direct detection of viral DNA and RNA, respectively. The reaction mixtures were partitioned with a common vortex mixer to generate picoliter-scale polydisperse droplets in several seconds. The limit of detection (LoD) for the target DNA and RNA is approximately 100 aM and 10 aM, respectively, which is about 3 × 104-105 fold more sensitive than corresponding bulk CRISPR assays. We applied the PddCas to successfully detect severe acute respiratory syndrome coronavirus (SARS-CoV-2) and human papillomavirus type 18 (HPV 18) in clinical samples. For the 23 HPV 18-suspected cervical epithelial cell samples and 32 nasopharyngeal swabs for SARS-CoV-2, 100% sensitivity and 100% specificity were demonstrated. The dual-gene virus detection with PddCas was also established and verified. Therefore, PddCas has potential for point-of-care application and is envisioned to be readily deployed for frequent testing as part of an integrated public health surveillance program.

Contrast-enhanced US with Perfluorobutane to Diagnose Small Lateral Cervical Lymph Node Metastases of Papillary Thyroid Carcinoma.

Background Although metastasis of lateral cervical lymph nodes (LNs) is common in papillary thyroid carcinoma (PTC), correctly diagnosing small metastatic LNs with US remains challenging. The use of contrast-enhanced US (CEUS), especially the postvascular phase of CEUS with perfluorobutane contrast material, might contribute to improved diagnosis of metastatic LNs in PTC. Purpose To assess the diagnostic value of the postvascular phase of CEUS with perfluorobutane in suspicious small lateral cervical LNs (short-axis diameter ≤8 mm) in patients with PTC. Materials and Methods This single-center study prospectively enrolled consecutive participants with confirmed PTC and suspicious LNs at US from October 2020 to October 2021. All participants underwent CEUS, 1 week before biopsy or surgery, with intravenous perfluorobutane contrast material to visualize the LNs in the vascular phase (5-60 seconds after injection) and postvascular phase (10-30 minutes after injection). The reference standard was cytologic and surgical histologic assessment of the LNs. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of sonographic features were calculated, and the diagnostic performance of US, CEUS, and the combination of postvascular phase and US features was assessed using multivariable logistic regression models. Results A total of 135 participants (median age, 36 years [IQR, 30-46 years]; 100 women) with 161 suspicious LNs at US were evaluated, including 67 metastatic and 94 benign LNs. The specificity of perfusion defect as a sonographic feature in the vascular phase reached 96% (90 of 94 LNs), and the negative predictive value of non-isoenhancement (ie, hypoenhancement, partial enhancement, and no enhancement) in the postvascular phase reached 100% (83 of 83 LNs). The area under the receiver operating characteristic curve (AUC) of the combination of postvascular phase and US features was 0.94 (95% CI: 0.89, 0.97), significantly higher than that of US features alone (AUC, 0.73; 95% CI: 0.65, 0.79; P < .001). Conclusion The postvascular phase of CEUS with perfluorobutane demonstrated excellent performance for diagnosing suspicious small lateral cervical LNs in participants with PTC. Published under a CC BY 4.0 license Supplemental material is available for this article. See also the editorial by Gunabushanam in this issue.

Contrast-enhanced US with Sulfur Hexafluoride and Perfluorobutane: LI-RADS for Diagnosing Hepatocellular Carcinoma.

Background Liver Imaging Reporting and Data System (LI-RADS) was designed for contrast-enhanced US (CEUS) with pure blood pool agents to diagnose hepatocellularfcarcinoma (HCC), such as sulfur hexafluoride (SHF), but Kupffer-cell agents, such as perfluorobutane (PFB), allow additional lesion characterization in the Kupffer phase yet remain unaddressed. Purpose To compare the diagnostic performance of three algorithms for HCC diagnosis: two algorithms based on CEUS LI-RADS version 2017 for both SHF and PFB and a modified algorithm incorporating Kupffer-phase findings for PFB. Materials and Methods This multicenter prospective study enrolled high-risk patients for HCC from June 2021 to December 2021. Each participant underwent same-day SHF-enhanced US followed by PFB-enhanced US. Each liver observation was assigned three LI-RADS categories according to each algorithm: LI-RADS SHF, LI-RADS PFB, and modified PFB. For modified PFB, observations at least 10 mm with nonrim arterial phase hyperenhancement were upgraded LR-4 to LR-5 if there was no washout with a Kupffer defect and were reassigned LR-M to LR-5 if there was early washout with mild Kupffer defect. The reference standard was pathologic confirmation or composite (typical CT or MRI features, or 1-year size stability and/or reduction). Diagnostic metrics of LR-5 for HCC using the three algorithms were calculated and compared using the McNemar test. Results Overall, 375 patients (mean age, 56 years ± 11 [SD]; 318 male patients, 57 female patients) with 424 observations (345 HCCs, 40 non-HCC malignancies, 39 benign lesions) were enrolled. PFB and SHF both using LI-RADS showed no significant difference in sensitivity (60% vs 58%; P = .41) and specificity (96% vs 95%; P > .99). The modified algorithm with PFB had increased sensitivity (80% vs 58%; P < .001) and a nonsignificant decrease in specificity (92% vs 95%; P = .73) compared with LI-RADS SHF. Conclusion Based on CEUS LI-RADS version 2017, both SHF and PFB achieved high specificity and relatively low sensitivity for HCC diagnosis. When incorporating Kupffer-phase findings, PFB had higher sensitivity without loss of specificity. Chinese Clinical Trial Registry no. ChiCTR2100047035 © RSNA, 2023 Supplemental material is available for this article. See also the editorial by Kim in this issue.

Long-Term Retention Microbubbles with Three-Layer Structure for Floating Intravesical Instillation Delivery.

Intravesical instillation is an effective treatment for bladder cancer. However, clinical anticancer agents always suffer rapid excretion by periodic urination, leading to low therapeutic efficacy. Prolonging the retention time of drugs in the bladder is the key challenge for intravesical instillation treatment. Herein, a facile and powerful surface cross-linking-freeze drying strategy is proposed to generate ultra-stable albumin bovine air microbubbles (BSA-MBs) that can float and adhere to the bladder wall to overcome the excretion of urination and exhibit a remarkable property of long-term retention in the bladder. More noteworthy, BSA-MBs are endowed with a specific three-layer structure, namely, the outer membrane, middle drug loading layer and inner air core, which makes them have a low density to easily float and possess a high drug loading capacity. Based on their unique superiorities, the therapeutic potential of doxorubicin (DOX)-loaded BSA-MBs (DOX-MBs) is exemplified by intravesical instillation for bladder cancer. After injection into the bladder, DOX-MBs can remain in the bladder for a long time and sustain the release of DOX in urine, exhibiting potent anticancer efficacy. Consequently, the prolonged retention of BSA-MBs in the bladder renders them as an effective floating drug delivery system for intravesical instillation therapy.

Peste Des Petits Ruminants Virus N Protein Is a Critical Proinflammation Factor That Promotes MyD88 and NLRP3 Complex Assembly.

Inflammatory responses play a central role in host defense against invading pathogens. Peste des petits ruminants virus (PPRV) causes highly contagious acute or subacute disease of small ruminants. However, the precise mechanism by which PPRV regulates inflammatory responses remains unknown. Here, we revealed a novel mechanism by which PPRV induces inflammation. Our study showed that PPRV induced the secretion of interleukin 1ß (IL-1ß) by activating the NF-κB signaling pathway and the NLRP3 inflammasome. Moreover, PPRV replication and protein synthesis were essential for NLRP3 inflammasome activation. Importantly, PPRV N protein promoted NF-κB signaling pathway and NLRP3 inflammasome via direct binding of MyD88 and NLPR3, respectively, and induced caspase-1 cleavage and IL-1ß maturation. Biochemically, N protein interacted with MyD88 to potentiate the assembly of MyD88 complex and interacted with NLPR3 to facilitate NLRP3 inflammasome complex assembly by forming an N-NLRP3-ASC ring-like structure, leading to IL-1ß secretion. These findings demonstrate a new function of PPRV N protein as an important proinflammation factor and identify a novel underlying mechanism modulating inflammasome assembly and function induced by PPRV. IMPORTANCE An important part of the innate immune response is the activation of NF-κB signaling pathway and NLPR3 inflammasome, which is induced upon exposure to pathogens. Peste des petits ruminants virus (PPRV) is a highly contagious virus causing fever, stomatitis, and pneumoenteritis in goats by inducing many proinflammatory cytokines. Although the NF-κB signaling pathway and NLRP3 inflammasome play an important role in regulating host immunity and viral infection, the precise mechanism by which PPRV regulates inflammatory responses remains unknown. This study demonstrates that PPRV induces inflammatory responses. Mechanistically, PPRV N protein facilitates the MyD88 complex assembly by directly binding to MyD88 and promotes the NLRP3 inflammasome complex assembly by directly binding to NLRP3 to form ring-like structures of N-NLRP3-ASC. These findings provide insights into the prevention and treatment of PPRV infection.

Using ultrasound radiomics analysis to diagnose cervical lymph node metastasis in patients with nasopharyngeal carcinoma.

OBJECTIVE: This study aimed to explore the clinical value of ultrasound radiomics analysis in the diagnosis of cervical lymph node metastasis (CLNM) in patients with nasopharyngeal carcinoma (NPC). METHODS: A total of 205 cases of NPC CLNM and 284 cases of benign lymphadenopathy with pathologic diagnosis were retrospectively included. Grayscale ultrasound (US) images of the largest section of every lymph node underwent feature extraction. Feature selection was done by maximum relevance minimum redundancy (mRMR) algorithm and multivariate logistic least absolute shrinkage and selection operator (LASSO) regression. Logistic regression models were developed based on clinical features, radiomics features, and the combination of those features. The AUCs of models were analyzed by DeLong's test. RESULTS: In the clinical model, lymph nodes in the upper neck, larger long axis, and unclear hilus were significant factors for CLNM (p < 0.001). MRMR and LASSO regression selected 7 significant features for the radiomics model from the 386 radiomics features extracted. In the validation dataset, the AUC value was 0.838 (0.776-0.901) in the clinical model, 0.810 (0.739-0.881) in the radiomics model, and 0.880 (0.826-0.933) in the combined model. There was not a significant difference between the AUCs of clinical models and radiomics models in both datasets. DeLong's test revealed a significantly larger AUC in the combined model than in the clinical model in both training (p = 0.049) and validation datasets (p = 0.027). CONCLUSION: Ultrasound radiomics analysis has potential value in screening meaningful ultrasound features and improving the diagnostic efficiency of ultrasound in CLNM of patients with NPC. KEY POINTS: • Radiomics analysis of gray-scale ultrasound images can be used to develop an effective radiomics model for the diagnosis of cervical lymph node metastasis in nasopharyngeal carcinoma patients. • Radiomics model combined with general ultrasound features performed better than the clinical model in differentiating cervical lymph node metastases from benign lymphadenopathy.

Predicting symptomatic post-hepatectomy liver failure in patients with hepatocellular carcinoma: development and validation of a preoperative nomogram.

OBJECTIVE: To develop and validate a nomogram based on liver stiffness (LS) for predicting symptomatic post-hepatectomy (PHLF) in patients with hepatocellular carcinoma (HCC). METHODS: A total of 266 patients with HCC were enrolled prospectively from three tertiary referral hospitals from August 2018 to April 2021. All patients underwent preoperative laboratory examination to obtain parameters of liver function. Two-dimensional shear wave elastography (2D-SWE) was performed to measure LS. Three-dimensional virtual resection obtained the different volumes including future liver remnant (FLR). A nomogram was developed by using logistic regression and determined by receiver operating characteristic (ROC) curve analysis and calibration curve analysis, which was validated internally and externally. RESULTS: A nomogram was constructed with the following variables: FLR ratio (FLR of total liver volume), LS greater than 9.5 kPa, Child-Pugh grade, and the presence of clinically significant portal hypertension (CSPH). This nomogram enabled differentiation of symptomatic PHLF in the derivation cohort (area under curve [AUC], 0.915), internal fivefold cross-validation (mean AUC, 0.918), internal validation cohort (AUC, 0.876) and external validation cohort (AUC, 0.845). The nomogram also showed good calibration in the derivation, internal validation, and external validation cohorts (Hosmer-Lemeshow goodness-of-fit test, p = 0.641, p = 0.06, and p = 0.127, respectively). Accordingly, the safe limit of the FLR ratio was stratified using the nomogram. CONCLUSION: An elevated level of LS was associated with the occurrence of symptomatic PHLF in HCC. A preoperative nomogram integrating LS, clinical and volumetric features was useful in predicting postoperative outcomes in patients with HCC, which might help surgeons in the management of HCC resection. CLINICAL RELEVANCE STATEMENT: A serial of the safe limit of the future liver remnant was proposed by a preoperative nomogram for hepatocellular carcinoma, which might help surgeons in 'how much remnant is enough in liver resection'. KEY POINTS: • An elevated liver stiffness with the best cutoff value of 9.5 kPa was associated with the occurrence of symptomatic post-hepatectomy liver failure in hepatocellular carcinoma. • A nomogram based on both quality (Child-Pugh grade, liver stiffness, and portal hypertension) and quantity of future liver remnant was developed to predict symptomatic post-hepatectomy liver failure for HCC, which enabled good discrimination and calibration in both derivation and validation cohorts. • The safe limit of future liver remnant volume was stratified using the proposed nomogram, which might help surgeons in the management of HCC resection.

Intraindividual Comparison of Contrast-Enhanced Ultrasound Using Perfluorobutane With Modified Criteria Versus CT/MRI LI-RADS Version 2018 for Diagnosing HCC in High-Risk Patients.

BACKGROUND. Previously proposed modifications to LI-RADS criteria for contrast-enhanced ultrasound (CEUS) performed using perfluorobutane contrast agent yielded increased sensitivity for hepatocellular carcinoma (HCC) without a significant decrease in specificity. OBJECTIVE. The purpose of our study was to compare the diagnostic performance of CEUS with perfluorobutane using modified LI-RADS criteria versus contrast-enhanced CT or MRI using LI-RADS version 2018 (v2018) for characterizing lesions as HCC in high-risk patients. METHODS. This retrospective study included 171 patients (140 men, 31 women; mean age, 54 ± 12 [SD] years) at high-risk for HCC with a pathologically confirmed liver observation evaluated by both CEUS using perfluorobutane and contrast-enhanced CT or MRI between March 2020 and May 2021. A matching algorithm was used to select two patients with HCC for each patient with a non-HCC lesion. Two readers evaluated observations using previously proposed modifications to CEUS LI-RADS version 2017 that classify certain observations as LR-5 rather than as LR-4 or LR-M on the basis of the presence of Kupffer phase defect after perfluorobutane administration; two different readers evaluated observations using CT/MRI LI-RADS v2018. Each reader pair reached consensus. Diagnostic performance was evaluated. RESULTS. A total of 114 patients had HCC, 43 had a non-HCC malignancy, and 14 had a benign lesion. Modified CEUS criteria using perfluorobutane and CT/MRI LI-RADS v2018 showed no significant difference (p > .05) in sensitivity (92.1% vs 89.5%), specificity (87.7% vs 84.2%), or accuracy (90.6% vs 87.7%) of LR-5 for diagnosis of HCC. Of six observations assessed as LR-4 only by CT/MRI LI-RADS v2018, modified CEUS criteria using perfluorobutane assessed one as LR-3 (benign lesion) and five as LR-5 (all HCC). Of seven observations assessed as LR-M only by CT/MRI LI-RADS v2018, modified CEUS criteria using perfluorobutane assessed one as LR-3 (non-HCC malignancy) and six as LR-5 (all HCC). Eight of 12 observations assessed as LR-5 only by CT/MRI LI-RADS v2018 and 11 of 13 observations assessed as LR-5 only by modified CEUS criteria using perfluorobutane were HCC. CONCLUSION. The diagnostic performance of LR-5 for HCC diagnosis was not significantly different between modified CEUS criteria using perfluorobutane and CT/MRI LI-RADS v2018. CLINICAL IMPACT. The findings support the application of modified CEUS criteria using perfluorobutane for diagnosing HCC in high-risk patients.

The oral bacterial microbiota facilitates the stratification for ulcerative colitis patients with oral ulcers.

BACKGROUND: Clinically, a large part of inflammatory bowel disease (IBD) patients is complicated by oral lesions. Although previous studies proved oral microbial dysbiosis in IBD patients, the bacterial community in the gastrointestinal (GI) tract of those IBD patients combined with oral ulcers has not been profiled yet. METHODS: In this study, we enrolled four groups of subjects, including healthy controls (CON), oral ulcer patients (OU), and ulcerative colitis patients with (UC_OU) and without (UC) oral ulcers. Bio-samples from three GI niches containing salivary, buccal, and fecal samples, were collected for 16S rRNA V3-V4 region sequencing. Bacterial abundance and related bio-functions were compared, and data showed that the fecal microbiota was more potent than salivary and buccal microbes in shaping the host immune system. ~ 22 UC and 10 UC_OU 5-aminosalicylate (5-ASA) routine treated patients were followed-up for six months; according to their treatment response (a decrease in the endoscopic Mayo score), they were further sub-grouped as responding and non-responding patients. RESULTS: We found those UC patients complicated with oral ulcers presented weaker treatment response, and three oral bacterial genera, i.e., Fusobacterium, Oribacterium, and Campylobacter, might be connected with treatment responding. Additionally, the salivary microbiome could be an indicator of treatment responding in 5-ASA routine treatment rather than buccal or fecal ones. CONCLUSIONS: The fecal microbiota had a strong effect on the host's immune indices, while the oral bacterial microbiota could help stratification for ulcerative colitis patients with oral ulcers. Additionally, the oral microbiota had the potential role in reflecting the treatment response of UC patients. Three oral bacteria genera (Fusobacterium, Oribacterium, and Campylobacter) might be involved in UC patients with oral ulcers lacking treatment responses, and monitoring oral microbiota may be meaningful in assessing the therapeutic response in UC patients.

CKM and TERT dual promoters drive CRISPR-dCas9 to specifically inhibit the malignant behavior of osteosarcoma cells.

Improvements in treatment and chemotherapy have increased the survival rate of osteosarcoma, but overall efficacy remains low, highlighting the need for new gene therapy methods. Clustered regularly interspaced short palindromic repeats-deactivated Cas9 (CRISPR-dCas9) technology offers a promising strategy, but targeting osteosarcoma cells precisely is a challenge. We designed a system to achieve specific expression of CRISPR-dCas9-KRAB in osteosarcoma cells by using the creatine kinase muscle (CKM) promoter to drive dCas9-KRAB and the telomerase reverse transcriptase (TERT) promoter to drive single guide (sg)RNA expression. We inhibited the MDM2 proto-oncogene using this system in vitro, which efficiently inhibited the malignant behavior of osteosarcoma cells and induced apoptosis without affecting normal cells. In vivo experiments demonstrated that this system effectively inhibited the growth of subcutaneously transplanted tumors in nude mice. These findings provide a new method for precise identification and intervention of osteosarcoma with significant implications for the development of gene therapy methods for other cancers. Future research should focus on optimizing this system for clinical translation.