The deubiquitinase USP28 maintains the expression of the transcription factor MYCN and is essential in neuroblastoma cells.

Neuroblastoma (NB) is one of the most common extracranial solid tumors in children. MYCN gene amplification is highly associated with poor prognosis in high-risk NB patients. In non-MYCN-amplified high-risk NB patients, the expression of c-MYC (MYCC) and its target genes is highly elevated. USP28 as a deubiquitinase is known to regulate the stability of MYCC. We show here USP28 also regulates the stability of MYCN. Genetic depletion or pharmacologic inhibition of the deubiquitinase strongly destabilizes MYCN and stops the growth of NB cells that overexpress MYCN. In addition, MYCC could be similarly destabilized in non-MYCN NB cells by compromising USP28 function. Our results strongly suggest USP28 as a therapeutic target for NB with or without MYCN amplification/overexpression.

Artificial intelligence applications in pediatric oncology diagnosis.

Artificial intelligence (AI) algorithms have been applied in abundant medical tasks with high accuracy and efficiency. Physicians can improve their diagnostic efficiency with the assistance of AI techniques for improving the subsequent personalized treatment and surveillance. AI algorithms fundamentally capture data, identify underlying patterns, achieve preset endpoints, and provide decisions and predictions about real-world events with working principles of machine learning and deep learning. AI algorithms with sufficient graphic processing unit power have been demonstrated to provide timely diagnostic references based on preliminary training of large amounts of clinical and imaging data. The sample size issue is an inevitable challenge for pediatric oncology considering its low morbidity and individual heterogeneity. However, this problem may be solved in the near future considering the exponential advancements of AI algorithms technically to decrease the dependence of AI operation on the amount of data sets and the efficiency of computing power. For instance, it could be a feasible solution by shifting convolutional neural networks (CNNs) from adults and sharing CNN algorithms across multiple institutions besides original data. The present review provides important insights into emerging AI applications for the diagnosis of pediatric oncology by systematically overviewing of up-to-date literature.

Contrast computed tomography-based radiomics is correlation with COG risk stratification of neuroblastoma.

PURPOSE: Although a risk stratification strategy for neuroblastoma (NB) has been proposed, precise and convenient clinical risk estimation remains challenging. This study aimed to investigate the correlation of contrast computed tomography (CT)-based radiomics with NB risk stratification. METHODS: Patients with NB (n = 289) from two centers (244 and 45 patients in the training/testing and external validation cohorts, respectively) were divided into nonhigh- and high-risk groups. A total of 1648 radiomics features were extracted from the arterial phase, and the radiomics signature was constructed using rad scores, whereas the clinical model was established based on clinical factors. Further, a combined nomogram was developed based on the clinical factors and radiomics signatures. Finally, receiver operating characteristic curve and decision curve analyses (DCA) were used to assess the performance of the established models. RESULTS: Seventeen radiomics features were used to construct the radiomics signature. A significant difference was observed in the rad score between the two groups in the training (0.540 vs. 0.704, P < 0.001) and testing (0.563 vs. 0.969, P < 0.001) cohorts. The nomogram showed a higher area under the curve (AUC) in the training (AUC = 0.87), testing (AUC = 0.83), and external validation (AUC = 0.84) cohorts than other models. The Hosmer-Lemeshow test and calibration curves indicated that the nomogram fit perfectly. DCA demonstrated that the clinical-radiomics nomogram was more beneficial. CONCLUSIONS: Contrast CT-based radiomics shows correlation with COG risk stratification of NB. Radiomics features combined with clinical factors showed the best performance, which may improve the management of patients with NB.

A Novel Model for Predicting the Clearance of Jaundice in Patients With Biliary Atresia After Kasai Procedure.

BACKGROUND: The failed clearance of jaundice (CJ) in patients with biliary atresia (BA) after the Kasai procedure (KP) often leads to a shorter native liver survival (NLS) time and earlier liver transplantation. We aimed to investigate risk factors of failed CJ and establish a novel nomogram model to predict the status of CJ. METHODS: We retrospectively reviewed institutional medical records from January 2015 to April 2020 and enrolled BA patients post-KP, randomly divided into training and testing cohorts at a ratio of 7:3, and further subdivided into cleared and uncleared jaundice groups. Univariate and multiple logistic regression analyses were used to select risk factors to establish the nomogram in the training cohort. The performance of the nomogram was evaluated by calculating the areas under the receiver operating curve (AUC) in both cohorts. RESULTS: This study included 175 BA patients post-KP. After univariate and multiple logistic regression analyses, Cytomegalovirus IgM +ve associated BA (OR = 3.38; 95% CI 1.01-11.32; P = 0.04), ln γ-glutamyl transpeptidase (GGT) (OR = 0.41; 95% CI 0.22-0.80; P = 0.009), thickness of the fibrous portal plate (OR = 0.45; 95% CI 0.27-0.76; P = 0.003), liver stiffness measurement (LSM) (OR = 1.19; 95% CI 1.06-1.34; P = 0.002), and multiple episodes of cholangitis (OR = 1.65; 95% CI 1.13-2.41; P = 0.01) were identified as independent risk factors of unsuccessful CJ to construct the nomogram. The receiver operating characteristic curve (ROC) analysis suggested good nomogram performance in both the training (AUC = 0.96) and testing cohorts (AUC = 0.91). CONCLUSION: Our nomogram model including several risk factors effectively predicts CJ in patients post-KP, which could aid in clinical decision-making.

Sensory and Volatile Compounds Characteristics of the Sauce in Bean Paste Fish Treated with Ultra-High-Pressure and Representative Thermal Sterilization.

This study investigated the differences between three sterilized samples to reveal the unique aroma characteristics of the sauce in bean paste fish by multiple analysis methodologies. Samples were subjected to pasteurized (PS), high-temperature sterilization (HTS), and ultra-high-pressure treatment (UHP) tests. The UHP had a higher sensory evaluation and could better maintain the original flavor of the sample. A total of 92, 83, 85, and 76 volatile compounds were detected via comprehensive two-dimensional gas chromatography-mass spectrometry (GC×GC-MS) techniques in the control (CT), PS, HTS, and UHP groups, respectively. According to the analysis of gas chromatograph-olfactometry and odor activity value, 7 compounds were considered to have an aromatic influence on the sauces, in which four compounds (1,8-Cineole, Linalool, Hexanal, and Dimethyl trisulfide) exhibited a positive contribution to the aroma of the sauces. PLS-DA results showed that the UHP group positively correlated with volatiles (Isoamylol and 1-Octen-3-ol), color, and gloss. In general, the UHP treatment could retain the original state and flavor of the sauce, showing a high similarity to the control group. The HTS significantly altered the flavor and status of the samples.

Thoracoscopic lobectomy through the pulmonary hilum approach for the treatment of congenital lung malformation.

BACKGROUND: Thoracoscopic lobectomy is widely accepted for the treatment of congenital lung malformations (CLM), owing to its advantages. However, severe incomplete interlobar fissure may lead to a high rate of conversion to thoracotomy and postoperational complications. Thoracoscopic lobectomy utilizing the pulmonary hilum approach may be an effective and safe method to resolve these problems. This retrospective study was conducted to estimate the safety and efficiency of this approach. METHODS: A retrospective review of medical records was performed in our institution, from January 2014 to December 2019, and 432 patients with CLM who underwent thoracoscopic lobectomy through the pulmonary hilum approach were included in this study. Patients were divided into the incomplete fissure (IF) group and complete fissure (CF) group according to the level of fissure, which was proposed by an anatomical classification of pulmonary fissures. RESULTS: Patients in the IF and CF groups were 131 and 301, respectively. In univariate analysis, there were statistical significances between the two groups in terms of intraoperative blood loss (P = 0.04), surgical time (P = 0.01), the number of chest tube drainages (P < 0.01), and the total length of hospital stay (P = 0.03). However, no patients experienced bronchopleural fistula, postoperative pneumonia, or conversion to thoracotomy in either group. Five patients experienced prolonged air leakage in the IF group, and no prolonged air leakage occurred in the CF group. CONCLUSION: An IF would certainly increase the difficulty of CLM surgery, and thoracoscopic lobectomy using the pulmonary hilum approach is an effective and safe method for CLM patients.

A Report of 21 Cases of Laparoscopic Liver Resection in Children.

Objective: To summarize and analyze the technique and experience of laparoscopic liver resection (LLR) in children. Methods: From Dec 2015 to Dec 2018, surgical outcomes were evaluated in 30 pediatric patients with liver tumors. LLR was performed in 21 patients, including 15 males and 6 females with a mean age of 7.6 ± 3.8 years (range, 2-13 years) and a mean liver tumor diameter of 7.2 ± 2.0 cm (range, 4-10 cm). During the same period, 9 cases of traditional open liver resection were selected as controls, including 6 males and 3 females with a mean age of 7.0 ± 3.7 years (range, 2-13 years) and a mean liver tumor diameter of 7.5 ± 2.2 cm (range, 4.4-10.0 cm). Results: The mean operation time and mean hepatic inflow occlusion time were longer in the laparoscopic group, with values of 182.3 ± 66.1 minutes and 83.2 ± 38.2 minutes, respectively, than in the open group, with values of 120.0 ± 54.3 minutes and 49.6 ± 26.5 minutes, respectively. There were no significant differences between the two groups in blood loss (164.4 ± 107.4 mL versus 133.8 ± 87.1 mL), blood transfusion volume (102.2 ± 100.2 mL versus 69.2 ± 81.8 mL), and alanine aminotransferase levels 24 hours after surgery (212.0 ± 101.4 IU/L versus 173.3 ± 97.2 IU/L) and 72 hours after surgery (74.0 ± 32.5 IU/L versus 81.7 ± 57.3 IU/L). The average hospital stay in the laparoscopic group (6.2 ± 1.6 days) was significantly shorter than that in the open group (8.2 ± 2.0 days). The children in both groups recovered well after surgery without serious complications. Conclusion: LLR contributes to improved recovery of patients after surgery without increasing the incidence of complication, therefore, LLR is a safe option for liver tumors in children.

Risk factors and associated outcomes of early acute kidney injury in pediatric liver transplant recipients: A retrospective study.

BACKGROUND: Acute kidney injury (AKI) may contribute to high mortality rates after liver transplantation. Few studies have investigated AKI in pediatric liver transplantation. This retrospective study was conducted to investigate the risk factors for and associated outcomes of AKI in pediatric liver transplant recipients. METHODS: Eighty pediatric liver transplant patients were included. The occurrence of AKI was defined by the KDIGO Clinical Practice Guidelines for Acute Kidney Injury. A multivariate regression analysis model was used to investigate risk factors for AKI in the pediatric liver recipients. RESULTS: The final multivariable regression model showed that biliary atresia (odds ratio [OR] = 0.097, p = 0.03), increased time of the anhepatic phase (OR = 0.871, p = 0.005) and lower postoperative jaundice clearance (OR = 13.936, p = 0.02) were independently associated with the development of AKI in pediatric patients. Additionally, cumulative 3-year patient (p = 0.15) and graft (p = 0.26) survival rates between the non-acute kidney injury (NAKI) and AKI groups were 95.2% vs 86.8% and 90.5% vs 84.2%, respectively. CONCLUSION: Pediatric liver transplant recipients with a presence of biliary atresia, increased time of anhepatic phase, and a lower postoperative jaundice clearance had an increased risk of AKI. The long-term outcomes of patients who developed AKI appears to be worse compared with those having NAKI. TYPE OF STUDY: Prognosis study. LEVEL OF EVIDENCE: Level III.

Post-traumatic stress disorder in living donors after pediatric liver transplantation: A cross-sectional investigation study.

Liver transplantation can lead to post-traumatic stress disorder (PTSD) in recipients, but the risk factors associated with PTSD in living donors are unknown. To investigate this progression in pediatric living donors, a cross-sectional investigation was carried out.All participants completed 2 questionnaires: a PTSD self-rating scale (PTSD-SS) and a validated Chinese version of the Medical Outcomes Study Short Form-36 (SF-36). Clinical and demographic data were collected from medical records and self-report questionnaires. Univariate analysis was conducted to identify statistical differences.The prevalence of full PTSD (all symptom clusters) and partial PTSD (2 out of 3 symptom clusters) was 12.1% and 31.1%, respectively. Those with an educational status of elementary school (P = .001), who were donors to their children (P = .008), who were in the first 6 months after transplant (P < .001), or were involved in transplants where the recipients had severe complications (P = .02) were more likely to have higher PTSD-SS scores than other groups. The non-PTSD group had a higher health-related quality-of-life score compared with the full and partial PTSD groups in the domains of physical function, role-physical, bodily pain, general health, vitality, social functioning, role-emotional, and mental health. In addition, the occurrence of PTSD was related to a poorer quality of life.The occurrence of PTSD was common in living donors after pediatric liver transplantation. Those with a lower educational status, who were donors to their children, were in the first 6 months after transplant, or were involved in transplants where the recipients had severe complications were most likely to experience PTSD. Post-traumatic stress symptom severity was significantly associated with a poorer quality of life after transplant.

Influence of bioluminescence imaging dynamics by D-luciferin uptake and efflux mechanisms.

Bioluminescence imaging (BLI) detects light generated by luciferase-mediated oxidation of substrate and is used widely for evaluating transgene expression in cell-based assays and in vivo in relevant preclinical models. The most commonly used luciferase for in vivo applications is firefly luciferase (fLuc), for which D-luciferin serves as the substrate. We demonstrated previously that the expression of the ABCG2 efflux transporter can significantly reduce BLI signal output and that HhAntag-691 can inhibit the efflux of D-luciferin, thereby enhancing BLI signal. Here we show that an HhAntag-691-sensitive uptake mechanism facilitates the intracellular concentration of D-luciferin and that the BLI dynamics of different cell lines are coregulated by this uptake mechanism in conjunction with ABCG2-mediated efflux. After administration of D-luciferin, the HhAntag-691-sensitive uptake mechanism generates a rapid increase in BLI signal that decreases over time, whereas ABCG2-mediated efflux stably reduces signal output. We implicate SLC22A4 (OCTN1), a member of the organic cation/zwitterion uptake transporter family, as one potential mediator of the HhAntag-691-sensitive D-luciferin uptake. These findings provide insight into mechanisms that contribute to the cellular uptake kinetics and in vivo biodistribution of D-luciferin.

Identification of inhibitors of ABCG2 by a bioluminescence imaging-based high-throughput assay.

ABCG2 is a member of the ATP-binding cassette (ABC) family of transporters, the overexpression of which is associated with tumor resistance to a variety of chemotherapeutic agents. Accordingly, combining ABCG2 inhibitor(s) with chemotherapy has the potential to improve treatment outcome. To search for clinically useful ABCG2 inhibitors, a bioluminescence imaging (BLI)-based assay was developed to allow high-throughput compound screening. This assay exploits our finding that d-luciferin, the substrate of firefly luciferase (fLuc), is a specific substrate of ABCG2, and ABCG2 inhibitors block the export of d-luciferin and enhance bioluminescence signal by increasing intracellular d-luciferin concentrations. HEK293 cells, engineered to express ABCG2 and fLuc, were used to screen the Hopkins Drug Library that includes drugs approved by the Food and Drug Administration (FDA) as well as drug candidates that have entered phase II clinical trials. Forty-seven compounds showed BLI enhancement, a measure of anti-ABCG2 activity, of > or =5-fold, the majority of which were not previously known as ABCG2 inhibitors. The assay was validated by its identification of known ABCG2 inhibitors and by confirming previously unknown ABCG2 inhibitors using established in vitro assays (e.g., mitoxantrone resensitization and BODIPY-prazosin assays). Glafenine, a potent new inhibitor, also inhibited ABCG2 activity in vivo. The BLI-based assay is an efficient method to identify new inhibitors of ABCG2. As they were derived from a FDA-approved compound library, many of the inhibitors uncovered in this study are ready for clinical testing.

Hedgehog pathway inhibitor HhAntag691 is a potent inhibitor of ABCG2/BCRP and ABCB1/Pgp.

HhAntag691 (GDC-0449), a low-molecular weight inhibitor of the tumor-promoting hedgehog (Hh) signaling pathway, has been used to treat medulloblastoma in animal models and has recently entered clinical trials for a variety of solid tumors. Here, we show that HhAntag691 inhibits multiple ATP-binding cassette (ABC) transporters. ATP-binding cassette transporters are within a family of membrane proteins, the overexpression of which is associated with multidrug resistance, a major impediment to successful cancer treatment. HhAntag691 is a potent inhibitor of two ABC transporters, ABCG2/BCRP and ABCB1/Pgp, and is a mild inhibitor of ABCC1/MRP1. In ABCG2-overexpressing HEK293 cells, HhAntag691 increased retention of the fluorescent ABCG2 substrate BODIPY-prazosin and resensitized these cells to mitoxantrone, an antineoplastic ABCG2 substrate. In Madin-Darby canine kidney II cells engineered to overexpress Pgp or MRP1, HhAntag691 increased the retention of calcein-AM and resensitized them to colchicine. HhAntag691 also resensitized human non-small cell lung carcinoma cells NCI-H460/par and NCI-H460/MX20, which overexpress ABCG2 in response to mitoxantrone, to mitoxantrone, and to topotecan or SN-38. The IC(50) values of HhAntag691 for inhibition of ABCG2 and Pgp were approximately 1.4 and approximately 3.0 microM, respectively. Because ABC transporters are highly expressed at the blood-brain barrier and on many tumor cells, they contribute significantly to treatment failure of many types of cancer, particularly of those within the neuraxis. In addition to its effect on Hh signaling, the ability of HhAntag691 and related compounds to inhibit two key ABC transporters could contribute to their effectiveness in treating malignancies.

ABCG2/BCRP expression modulates D-Luciferin based bioluminescence imaging.

Bioluminescence imaging (BLI) is becoming indispensable to the study of transgene expression during development and, in many in vivo models of disease such as cancer, for high throughput drug screening in vitro. Because reaction of d-luciferin with firefly luciferase (fLuc) produces photons of sufficiently long wavelength to permit imaging in intact animals, use of this substrate and enzyme pair has become the method of choice for performing BLI in vivo. We now show that expression of the ATP-binding cassette (ABC) family transporter ABCG2/BCRP affects BLI signal output from the substrate d-luciferin. In vitro studies show that d-luciferin is a substrate for ABCG2/BCRP but not for the MDR1 P-glycoprotein (ABCB1/Pgp), multidrug resistance protein 1 (MRP1/ABCC1), or multidrug resistance protein 2 (MRP2/ABCC2). d-Luciferin uptake within cells is shown to be modulated by ABC transporter inhibitors, including the potent and selective ABCG2/BCRP inhibitor fumitremorgin C. Images of xenografts engineered to express transgenic ABCG2/BCRP, as well as xenografts derived from the human prostate cancer cell line 22Rv1 that naturally express ABCG2/BCRP, show that ABCG2/BCRP expression and function within regions of interest substantially influence d-luciferin-dependent bioluminescent output in vivo. These findings highlight the need to consider ABCG2/BCRP effects during d-luciferin-based BLI and suggest novel high throughput methods for identifying new ABCG2/BCRP inhibitors.