Discrimination between benign and malignant gallbladder lesions on enhanced CT imaging using radiomics.

BACKGROUND: Gallbladder cancer is a rare but aggressive malignancy that is often diagnosed at an advanced stage and is associated with poor outcomes. PURPOSE: To develop a radiomics model to discriminate between benign and malignant gallbladder lesions using enhanced computed tomography (CT) imaging. MATERIAL AND METHODS: All patients had a preoperative contrast-enhanced CT scan, which was independently analyzed by two radiologists. Regions of interest were manually delineated on portal venous phase images, and radiomics features were extracted. Feature selection was performed using mRMR and LASSO methods. The patients were randomly divided into training and test groups at a ratio of 7:3. Clinical and radiomics parameters were identified in the training group, three models were constructed, and the models' prediction accuracy and ability were evaluated using AUC and calibration curves. RESULTS: In the training group, the AUCs of the clinical model and radiomics model were 0.914 and 0.968, and that of the nomogram model was 0.980, respectively. There were statistically significant differences in diagnostic accuracy between nomograms and radiomics features (P <0.05). There was no significant difference in diagnostic accuracy between the nomograms and clinical features (P >0.05) or between the clinical features and radiomics features (P >0.05). In the testing group, the AUC of the clinical model and radiomics model were 0.904 and 0.941, and that of the nomogram model was 0.948, respectively. There was no significant difference in diagnostic accuracy between the three groups (P >0.05). CONCLUSION: It was suggested that radiomics analysis using enhanced CT imaging can effectively discriminate between benign and malignant gallbladder lesions.

Evaluation of treatment effect modification by biomarkers measured pre- and post-randomization in the presence of non-monotone missingness.

In vaccine studies, an important research question is to study effect modification of clinical treatment efficacy by intermediate biomarker-based principal strata. In settings where participants entering a trial may have prior exposure and therefore variable baseline biomarker values, clinical treatment efficacy may further depend jointly on a biomarker measured at baseline and measured at a fixed time after vaccination. This makes it important to conduct a bivariate effect modification analysis by both the intermediate biomarker-based principal strata and the baseline biomarker values. Existing research allows this assessment if the sampling of baseline and intermediate biomarkers follows a monotone pattern, i.e., if participants who have the biomarker measured post-randomization would also have the biomarker measured at baseline. However, additional complications in study design could happen in practice. For example, in a dengue correlates study, baseline biomarker values were only available from a fraction of participants who have biomarkers measured post-randomization. How to conduct the bivariate effect modification analysis in these studies remains an open research question. In this article, we propose approaches for bivariate effect modification analysis in the complicated sampling design based on an estimated likelihood framework. We demonstrate advantages of the proposed method over existing methods through numerical studies and illustrate our method with data sets from two phase 3 dengue vaccine efficacy trials.

An anatomical study of persistent trigeminal artery detected by computed tomography angiography and magnetic resonance angiography: proposal for a modified classification and a novel basilar artery grading system.

PURPOSE: The purpose of this study is to investigate the anatomical characteristics of persistent trigeminal artery (PTA) detected by computed tomography angiography (CTA) and magnetic resonance angiography (MRA), propose a modified classification and a novel grading system for basilar artery (BA). METHODS: Patients who underwent head CTA or MRA in our hospital between August 2014 and August 2022 were reviewed retrospectively. The prevalence, sex, and course of PTA were evaluated. PTA types were modified based on Weon's classification. Type I to IV were similar to those in Weon's classification except the presence of intermed fetal-type posterior cerebral artery (IF-PCA). Type V was the same as that in Weon's classification. Type VI included subtypes of VIa (concomitant IF-PCA based on type I to IV) and VIb (other variants). BA was assessed based on a scale of 0 to 5 compared with PTA's caliber (0, BA aplasia; 1 and 2, BA non-dominant; 3, equilibrium; 4 and 5, BA dominant). RESULTS: A total of 57 patients (0.06%) with PTA, including 36 females and 21 males, were detected in 94,487 patients. Six patients (10.5%) were medial type and 51 patients (89.5%) were lateral type. Thirty-seven patients (64.9%) were type I, 1 (1.8%) as type II, 13 (22.8%) as type III, 3 (5.3%) as type IV, 1 (1.8%) as type V, and 2 (3.5%) as type VI. For BA grading, 4 (7.0%), 21 (36.8%), 17 (29.8%), 6 (10.5%), 6 (10.5%), and 3 (5.3%) of the patients were grade 0, 1, 2, 3, 4, and 5, respectively. Fifteen patients (26.3%) had intracranial aneurysms. One cases (1.8%) had a fenestration of the PTA. CONCLUSION: The prevalence of PTA in our study was lower than that in most previous reports. The modified PTA classification and BA grading system can be used to better understand the vascular structure of PTA patients.

Neutralizing Antibody Correlates Analysis of Tetravalent Dengue Vaccine Efficacy Trials in Asia and Latin America.

Background: In the CYD14 and CYD15 Phase 3 trials of the CYD-TDV dengue vaccine, estimated vaccine efficacy (VE) against symptomatic, virologically confirmed dengue (VCD) occurring between months 13 and 25 was 56.5% and 60.8%, respectively. Methods: Neutralizing antibody titers to the 4 dengue serotypes in the CYD-TDV vaccine insert were measured at month 13 in a randomly sampled immunogenicity subcohort and in all VCD cases through month 25 (2848 vaccine, 1574 placebo) and studied for their association with VCD and with the level of VE to prevent VCD. Results: For each trial and serotype, vaccinees with higher month 13 titer to the serotype had significantly lower risk of VCD with that serotype (hazard ratios, 0.19-0.43 per 10-fold increase). Moreover, for each trial, vaccinees with higher month 13 average titer to the 4 serotypes had significantly higher VE against VCD of any serotype (P < .001). Conclusions: Neutralizing antibody titers postdose 3 correlate with CYD-TDV VE to prevent dengue. High titers associate with high VE for all serotypes, baseline serostatus groups, age groups, and both trials. However, lowest titers do not fully correspond to zero VE, indicating that other factors influence VE.

Increased serum amyloid A in nasal polyps is associated with systemic corticosteroid insensitivity in patients with chronic rhinosinusitis with nasal polyps: a pilot study.

BACKGROUND: Serum amyloid A (SAA) was involved in the pathogenesis of glucocorticoid resistance in lung diseases. However, their association with systemic corticosteroid insensitivity in chronic rhinosinusitis with nasal polyps (CRSwNP) patients remains to be assessed. METHODS: This study enrolled 32 CRSwNP patients to evaluate the association between SAA expression in NP and corticosteroid insensitivity, and the value of polyp SAA level for predicting the response to oral corticosteroids in CRSwNP patients. All patients were given a course of oral prednisone (30 mg daily for 2 weeks) and subdivided into glucocorticoid(GC)-sensitive and -insensitive subgroup according to the change in polyp size scores. The polyp specimens were obtained before and after corticosteroid treatment. SAA levels in polyp tissues were evaluated by enzyme-linked immunosorbent assay and quantitative reverse transcription polymerase chain reaction. Regression analysis was performed to analyze the association between SAA protein levels and corticosteroid insensitivity. RESULTS: 13/32 (40.62%) CRSwNP patients were insensitive to the oral corticosteroid therapy. SAA mRNA and protein levels were significantly increased in GC-insensitive NP compared to those in GC-sensitive NP. Tissue SAA protein levels were positively correlated with tissue neutrophil numbers. Regression analysis revealed tissue SAA levels were significantly correlated with corticosteroid insensitivity (P < 0.01). ROC curves indicated that the area under the curve was 0.87. When the polyp SAA protein level was 122.2 ng/ml or higher, the sensitivity and specificity were 76.92 and 73.68%, respectively. CONCLUSIONS: Our findings suggest that increased SAA in NP is associated with reduced response to oral corticosteroids in CRSwNP. SAA levels in NP may have potential value in predicting corticosteroid insensitivity in CRSwNP patients.

Atypical choroid plexus papilloma: clinicopathological and neuroradiological features.

Background Atypical choroid plexus papilloma (APP) is a rare, newly introduced entity with intermediate characteristics. To date, few reports have revealed the magnetic resonance (MR) findings. Purpose To analyze the clinicopathological and MR features of APP. Material and Methods The clinicopathological data and preoperative MR images of six patients with pathologically proven APP were retrospectively reviewed. The MR features including tumor location, contour, signal intensity, degree of enhancement, intratumoral cysts, and necrosis; and flow voids, borders, peritumoral edema, and associated hydrocephalus were analyzed. Results The APP were located in the ventricle (n = 4) and cerebellopontine angle (CPA, n = 2). Tumor dissemination along the spinal subarachnoid space was found in one patient. The tumors appeared as milt-lobulated (n = 5) or round mass (n = 1), with slightly heterogeneous signals (n = 5) or mixed signals (n = 1) on T1-weighted and T2-weighted images. Heterogeneous and strong enhancement were found in five cases on contrast-enhanced images. Three of four intraventricular tumors had a partly blurred border with ventricle wall. Four tumors had mild to moderate extent of surrounding edema signals. A slight hydrocephalus was seen in four patients. Incomplete capsule was seen in four tumors at surgery. Histopathologically, mild nuclear atypia was seen in all tumors with a mitotic rate of 2-5 per 10 high-power fields. Conclusion APP should be included in the differential diagnosis when an intraventricular or CPA tumor appearing as a multi-lobulated solid mass with slight heterogeneity, heterogeneous strong enhancement, partly blurred borders, mild to moderate peritumoral edema, or slight hydrocephalus are present.

Photostability study of cis-configuration neonicotinoid insecticide cycloxaprid in water.

Cycloxaprid (CYC) is a new cis-configuration neonicotinoid insecticide, which is currently under development in China for agricultural pest control. Considering the photodegradation of CYC is important for the application of CYC in the future, the photochemical behavior of CYC in aqueous solution was herein investigated in a "merry-go-round" reactor under a 300 W high-pressure mercury lamp. Twenty-five photodegradation products were identified via UPLC-TOF-ESI-MS/MS. The results suggested that NTN32692, the precursor of CYC was the predominant photodegradation product. CYC photodegrades via a more complex mechanism than imidacloprid and four potential photodegradation pathways were proposed.

Application of Y-Z deformable magnetic ring for recanalization of transanal single-access rectal stricture.

Magnetic compression anastomosis has been reported to have remarkable clinical outcomes. Here, we tested the applicability of a Y-Z deformable magnetic ring (DMR) for non-surgical manipulation of rectal stenosis (RS) in a beagle dog model under a transanal single-access condition. RS was modeled in 8 beagle dogs using partial ligation with silk thread. Under X-ray guidance, the Y-Z DMR was positioned at the proximal and distal ends of the RS, and the magnetic ring was bent into an "O" shape, such that the two rings were magnetically attracted. Operation time, complications during or after operation, and discharge time of the magnetic rings were recorded. The anastomosis bursting pressure was measured two weeks after removing the rings, and its formation was assessed through gross and histological examination. Partial ligation with a silk thread successfully established the canine RS model. After Y-Z DMR installation, the magnetic ring was successfully reconfigured from an "S" to an "O" shape. Strong attraction existed between the rings. The operation time was 9-15 min (average: 11.75 ± 1.98 min). No rectal bleeding or perforation occurred during or after operation. The ring was naturally expelled 7-10 days after surgery. A pressure of > 300 mmHg was recorded at the point of anastomosis rupture. The rectal anastomosis appeared to have healed properly on the surface, which was confirmed histologically, signifying the success of this procedure. A Y-Z DMR facilitated the successful recanalization of transanal single-channel RS without needing surgery in an animal model.

A novel magnetic compression technique for establishment of a vesicovaginal fistula model in Beagle dogs.

Vesicovaginal fistula lacks a standard, established animal model, making surgical innovations for this condition challenging. Herein, we aimed to non-surgically establish vesicovaginal fistula using the magnetic compression technique, and the feasibility of this method was explored using eight female Beagle dogs as model animals. In these dogs, cylindrical daughter and parent magnets were implanted into the bladder and vagina, respectively, after anesthesia, and the positions of these magnets were adjusted under X-ray supervision to make them attract each other, thus forming the structure of daughter magnet-bladder wall-vaginal wall-parent magnet. Operation time and collateral damage were recorded. The experimental animals were euthanized 2 weeks postoperatively, and the vesicovaginal fistula gross specimens were obtained. The size of the fistula was measured. Vesicovaginal fistula was observed by naked eye and under a light microscope. Magnet placement was successful in all dogs, and remained in the established position for the reminder of the experiment. The average operation time was 14.38 min ± 1.66 min (range, 12-17 min). The dogs were generally in good condition postoperatively and were voiding normally, with no complications like bleeding and urine retention. The magnets were removed from the vagina after euthanasia. The vesicovaginal fistula was successfully established according to gross observation, and the fistula diameters were 4.50-6.24 mm. Histological observation revealed that the bladder mucosa and vaginal mucosa were in close contact on the internal surface of the fistula. Taken together, magnetic compression technique is a simple and feasible method to establish an animal model of vesicovaginal fistula using Beagle dogs. This model can help clinicians study new surgical techniques and practice innovative approaches for treating vesicovaginal fistula.

Post-operative cognitive dysfunction in patients following gastrointestinal endoscopic treatment.

We aimed to explore the changes in post-operative cognitive dysfunction (POCD) after gastrointestinal endoscopic treatment using detailed neuropsychological assessments. Patients hospitalized for gastrointestinal endoscopic polypectomy were recruited for neuropsychological evaluations, which included the Chinese version of the Mini-Mental State Examination (MMSE), Auditory-Verbal Learning Test, Digit Span Test (DST), Trail Making Task (TMT), Verbal Fluency Test, Clock Drawing Test, and Stroop test. Cognitive assessments were performed twice: one day before and 24 h after treatment. Healthy control subjects participated in the neuropsychological assessment during the same intervals. Detailed cognitive assessments were performed for 40 patients and 60 control subjects. Based on the Z scores, the incidence of POCD 24 h after gastrointestinal endoscopic treatment was 20%. Patients with POCD had significant impairment in the post-operative MMSE, forward DST, TMT, and Stroop interference effect correct count tests (all p < 0.05). Our preliminary results showed that patients were not fully recovered, and 20% had impairment in multiple cognitive assessments 24 h after a gastrointestinal endoscopy. As attention was affected, safety while discharging those patients should be a concern.

Altered resting-state hippocampal functional connectivity in breast cancer survivors with chemotherapy-induced amenorrhea.

INTRODUCTION: Amenorrhea induced decrease of hormones is associated with cognitive impairment. This study aimed to evaluate hippocampal functional connectivity patterns in chemotherapy-induced amenorrhea (CIA) breast cancer (BC) patients, to evaluate the relationship between the functional connectivity features and hormone levels. METHOD: Neuropsychological test, functional magnetic resonance imaging, and assessment of hormone levels were conducted in 21 premenopausal BC patients before chemotherapy (t0 ) and 1 week after completing chemotherapy (t1 ). Twenty matched healthy controls (HC) were also included and underwent the same assessments at similar time intervals. Mixed effect analysis and paired t-test were used to compare differences in brain functional connectivity. RESULTS: Voxel-based paired t-tests revealed increased functional connectivity of the right and left hippocampus with the left fusiform gyrus, inferior and middle temporal gyrus, inferior occipital gyrus, left lingual gyrus, and parahippocampal gyrus after chemotherapy (p < .001) in CIA patients. Repeated measures analysis revealed significant group-by-time interactions in the left hippocampus with the bilateral fusiform gyrus, right parahippocampal gyrus, left inferior temporal gyrus, and left inferior occipital gyrus (p < .001). Premenopausal BC patients had no significant differences in cognitive function compared with HC at baseline. However, the CIA patients had high levels of self-rating depression scale, self-rating anxiety scale, total cholesterol, and triglycerides. Further, the CIA patients showed significant differences in hormone and fasting plasma glucose levels and cognitive performances between t0 and t1 (p < .05). Functional connectivity changes between the left hippocampus and the left inferior occipital gyrus was negatively correlated with E2 and luteinizing hormone changes (p < .05). CONCLUSION: The CIA patients had cognitive dysfunction mainly in memory and visual mobility. Chemotherapy may affect hippocampal-posterior cortical circuit which mediates visual processing in CIA patients. Moreover, E2 may be involved in this process.

CircRNA ACVR2A Sponges miR-1290 to Modulate Cell Progression in Gastric Cancer.

BACKGROUND: In recent years, the abnormal expression of circRNAs has been identified to be strongly associated with tumor tissues. In this study, we focused on circACVR2A with a remarkably upregulated expression in gastric tissues and further explored its role in the pathogenic progression of gastric cancer (GC). METHODS: The differentially expressed circACVR2A in GC tissues and four cell lines (MKN-45, SNU-1, HGC-27, and SGC-7901) was identified by qRT-PCR method. Then, the effect of circACVR2A and miR-1290 on HGC-27 cell proliferation was measured by CCK8 and the colony formation methods. The effect of circACVR2A and miR-1290 on HGC-27 cell metastasis was estimated by transwell assay. The interaction of circACVR2A and miR-1290 was further detected. RESULTS: The relative level of circACVR2A in GC tissues and cell lines is remarkably upregulated. The downregulation of circACVR2A promotes GC cell proliferation and metastasis and suppressed the expression level of E-cadherin and Vimentin. The miR-1290 inhibitor reversed the effect of circACVR2A on cell progression in GC cell. CONCLUSION: circACVR2A competitively sponged miR-1290 and was exerted as a tumor suppressor gene oncogene via a circACVR2A/miR-1290 axis, suggesting it as a possible biomarker for GC therapy.

SENSITIVITY ANALYSIS FOR EVALUATING PRINCIPAL SURROGATE ENDPOINTS RELAXING THE EQUAL EARLY CLINICAL RISK ASSUMPTION.

This article addresses the evaluation of post-randomization immune response biomarkers as principal surrogate endpoints of a vaccine's protective effect, based on data from randomized vaccine trials. An important metric for quantifying a biomarker's principal surrogacy in vaccine research is the vaccine efficacy curve, which shows a vaccine's efficacy as a function of potential biomarker values if receiving vaccine, among an 'early-always-at-risk' principal stratum of trial participants who remain disease-free at the time of biomarker measurement whether having received vaccine or placebo. Earlier work in principal surrogate evaluation relied on an 'equal-early-clinical-risk' assumption for identifiability of the vaccine curve, based on observed disease status at the time of biomarker measurement. This assumption is violated in the common setting that the vaccine has an early effect on the clinical endpoint before the biomarker is measured. In particular, a vaccine's early protective effect observed in two phase III dengue vaccine trials (CYD14/CYD15) has motivated our current research development. We relax the 'equal-early-clinical-risk' assumption and propose a new sensitivity analysis framework for principal surrogate evaluation allowing for early vaccine efficacy. Under this framework, we develop inference procedures for vaccine efficacy curve estimators based on the estimated maximum likelihood approach. We then use the proposed methodology to assess the surrogacy of post-randomization neutralization titer in the motivating dengue application.

Concentration and distribution of sialic acid in human milk and its correlation with dietary intake.

Purpose: This study evaluates the content, distribution, and changing trend of sialic acid in human milk and the correlation between dietary intake of sialic acid and that in human milk. Methods: The study included 33 mothers of full-term and exclusively breastfed infants. At least 2 ml of milk was collected on the 3rd, 8th, 30th, and 90th day after delivery, and 24-h diet recalls of the lactating mothers were obtained each time. The correlation of human milk sialic acid concentration with lactating women's dietary sialic acid intake during lactation was analyzed by statistical analysis software SPSS. Results: The average concentration of sialic acid in colostrum, transition, and 1 and 3 months were 1,670.74 ± 94.53, 1,272.19 ± 128.74, 541.64 ± 55.2, and 297.65 ± 20.78 mg/L, respectively. The total sialic acid concentration in colostrum was about 5.6 times higher than that at 3 months (P < 0.001). The average dietary sialic acid intake of lactating mothers on the 2nd, 7th, 30th, and 90th day after delivery were 106.06 ± 7.51, 127.64 ± 8.61, 120.34 ± 10.21, and 95.40 ± 6.34 mg/day, respectively. The intake of sialic acid was relatively high on the 7th day, and there was no significant difference in dietary intake of sialic acid on different days (P < 0.05). In addition, there was no correlation between the intake of dietary sialic acid and the content of total sialic acid and various forms of sialic acid in milk (P < 0.05). Conclusion: During the lactation period, the distribution of sialic acid in breast milk is relatively stable and its content fluctuates greatly, which may not be affected by the mother's diet, but mainly depends on the self-regulation oft physiological needs.

Four circulating exosomal miRNAs as novel potential biomarkers for the early diagnosis of human colorectal cancer.

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths worldwide. Our study aimed to identify novel circulating miRNAs as early diagnostic biomarkers of CRC. The Gene Expression Omnibus (GEO) datasets were analyzed by using the online tool GEO2R. Isolated exosomes were verified by using the transmission electron microscope (TEM), Nanosight, and western blot. qRT-PCR was implemented to examine miRNA expression. The diagnostic value of circulating exosomal miRNAs was identified by using the receiver operating characteristic curve (ROC). In this study, we found that serum exosomal miRNAs are more suitable for diagnosing CRC when compared to serum miRNAs. Furthermore, we identified four exosomal miRNAs (miR-126, miR-1290, miR-23a, and miR-940) in the serum of CRC patients as novel potential biomarkers for the early diagnosis of CRC because they showed high diagnostic values to differentiate CRC patients at TNM stage I from healthy controls (HCs). In addition, our data suggested that CRC cells may secrete miRNAs into the extracellular environment through exosomes regardless of intracellular miRNA expression. In conclusion, we identified serum exosomal miR-126, miR-1290, miR-23a, and miR-940 as novel potential biomarkers for the early diagnosis of CRC.

Neurologic Factors in Patients with Vascular Mild Cognitive Impairment Based on fMRI.

This study focused on the application of functional magnetic resonance imaging and neuropsychology in diagnosis of vascular mild cognitive impairment (MCI) and the exploration of its relevant factors. The study enrolled 28 patients with vascular MCI in an observation group and 30 healthy individuals in a control group. All patients underwent magnetic resonance imaging. An automatic segmentation algorithm based on graph theory was adopted to process the images. Age, sex, disease course, Montreal Cognitive Assessment score, regional homogeneity, and amplitude of low-frequency fluctuation levels were recorded. There were no significant differences in age, gender, and course of disease between the observation group and the control group (P > 0.05). The level of regional homogeneity in the left posterior cerebellum in the observation group was significantly higher than that in the control group (P < 0.05).The regional homogeneity level of bilateral cingulate cortex was negatively correlated with Montreal Cognitive Assessment score (P < 0.05). The amplitude of low-frequency fluctuation of bilateral inferior parietal lobe, parietal lobe, and prefrontal lobe in the observation group was significantly lower than that in the control group, and the amplitude of low-frequency fluctuation of bilateral anterior cingulate gyrus, superior medial frontal gyrus, orbital frontal gyrus, right middle frontal gyrus, and right auxiliary motor area was higher than that in the control group (P < 0.05). Heart disease, such as myocardial infarction and atrial fibrillation, is a high risk factor for vascular MCI. Functional magnetic resonance imaging combined with an automatic segmentation algorithm can noninvasively observe the changes of a patient's brain tissue, which can be used in the recognition of vascular MCI. The global network attributes of patients with depression tend to be more randomized and have stronger resilience under targeted attacks.

Identification of Tumorigenic and Prognostic Biomarkers in Colorectal Cancer Based on microRNA Expression Profiles.

OBJECTIVE: Although noncoding RNAs, especially the microRNAs, have been found to play key roles in CRC development in intestinal tissue, the specific mechanism of these microRNAs has not been fully understood. METHODS: GEO and TCGA database were used to explore the microRNA expression profiles of normal mucosa, adenoma, and carcinoma. And the differential expression genes were selected. Computationally, we built the SVM model and multivariable Cox regression model to evaluate the performance of tumorigenic microRNAs in discriminating the adenomas from normal tissues and risk prediction. RESULTS: In this study, we identified 20 miRNA biomarkers dysregulated in the colon adenomas. The functional enrichment analysis showed that MAPK activity and MAPK cascade were highly enriched by these tumorigenic microRNAs. We also investigated the target genes of the tumorigenic microRNAs. Eleven genes, including PIGF, TPI1, KLF4, RARS, PCBP2, EIF5A, HK2, RAVER2, HMGN1, MAPK6, and NDUFA2, were identified to be frequently targeted by the tumorigenic microRNAs. The high AUC value and distinct overall survival rates between the two risk groups suggested that these tumorigenic microRNAs had the potential of diagnostic and prognostic value in CRC. CONCLUSIONS: The present study revealed possible mechanisms and pathways that may contribute to tumorigenesis of CRC, which could not only be used as CRC early detection biomarkers, but also be useful for tumorigenesis mechanism studies.

METTL14 Inhibits Hepatocellular Carcinoma Metastasis Through Regulating EGFR/PI3K/AKT Signaling Pathway in an m6A-Dependent Manner.

PURPOSE: Hepatocellular carcinoma (HCC) ranks as the fourth leading cause of cancer-related deaths worldwide. N6-methyladenosine (m6A) RNA methylation is the most common modification of messenger RNAs (mRNAs). The prognosis of HCC patients with metastasis remains poor. Our study aimed to elucidate the regulatory role of m6A on HCC metastasis. PATIENTS AND METHODS: All HCC patients were enrolled from The Affiliated Huai'an No. 1 People's Hospital of Nanjing Medical University. The expression levels of gene were tested by quantitative polymerase chain reaction (qPCR), Western blot, or immunohistochemistry (IHC) analysis. Wound healing assay, Transwell invasion assay, and lung metastasis model were implemented to investigate the migration and invasion ability of HCC cells. Candidate targets were selected by a comprehensive analysis of RNA-sequencing and m6A-sequencing of HepG2 cells. RESULTS: In this study, we demonstrated that METTL14 was significantly downregulated in HCC and significantly associated with the prognosis of HCC patients. METTL14 knockdown promoted the migration, invasion, and epithelial-mesenchymal transition (EMT) of HCC cells in vitro and in vivo. In addition, overlapping RNA-sequencing and m6A-sequencing data, we identified EGFR as a direct target of METTL14 in HCC. Mechanistically, METTL14 was found to inhibit HCC cell migration, invasion, and EMT through modulating EGFR/PI3K/AKT signaling pathway in an m6A-dependent manner. CONCLUSION: Targeting METTL14/EGFR/PI3K/AKT signaling pathway may facilitate the development of a new treatment strategy against the metastasis of HCC.

Microneutralization assay titer correlates analysis in two phase 3 trials of the CYD-TDV tetravalent dengue vaccine in Asia and Latin America.

We previously showed that Month 13 50% plaque reduction neutralization test (PRNT50) neutralizing antibody (nAb) titers against dengue virus (DENV) correlated with vaccine efficacy (VE) of CYD-TDV against symptomatic, virologically-confirmed dengue (VCD) in the CYD14 and CYD15 Phase 3 trials. While PRNT is the gold standard nAb assay, it is time-consuming and costly. We developed a next-generation high-throughput microneutralization (MN) assay and assessed its suitability for immune-correlates analyses and immuno-bridging applications. We analyzed MN and PRNT50 titers measured at baseline and Month 13 in a randomly sampled immunogenicity subset, and at Month 13 in nearly all VCD cases through Month 25. For each serotype, MN and PRNT50 titers showed high correlations, at both baseline and Month 13, with MN yielding a higher frequency of baseline-seronegatives. For both assays, Month 13 titer correlated inversely with VCD risk. Like PRNT50, high Month 13 MN titers were associated with high VE, and estimated VE increased with average Month 13 MN titer. We also studied each assay as a valid surrogate endpoint based on the Prentice criteria, which supported each assay as a valid surrogate for DENV-1 but only partially valid for DENV-2, -3, and -4. In addition, we applied Super-Learner to assess how well demographic, Month 13 MN, and/or Month 13 PRNT50 titers could predict Month 13-25 VCD outcome status; prediction was best when using demographic, MN, and PRNT50 information. We conclude that Month 13 MN titer performs comparably to Month 13 PRNT50 titer as a correlate of risk, correlate of vaccine efficacy, and surrogate endpoint. The MN assay could potentially be used to assess nAb titers in immunogenicity studies, immune-correlates studies, and immuno-bridging applications. Additional research would be needed for assessing the utility of MN titer in correlates analyses of other DENV endpoints and over longer follow-up periods.

Simultaneous Inference of Treatment Effect Modification by Intermediate Response Endpoint Principal Strata with Application to Vaccine Trials.

In randomized clinical trials, researchers are often interested in identifying an inexpensive intermediate study endpoint (typically a biomarker) that is a strong effect modifier of the treatment effect on a longer-term clinical endpoint of interest. Motivated by randomized placebo-controlled preventive vaccine efficacy trials, within the principal stratification framework a pseudo-score type estimator has been proposed to estimate disease risks conditional on the counter-factual biomarker of interest under each treatment assignment to vaccine or placebo, yielding an estimator of biomarker conditional vaccine efficacy. This method can be used for trial designs that use baseline predictors of the biomarker and/or designs that vaccinate disease-free placebo recipients at the end of the trial. In this article, we utilize the pseudo-score estimator to estimate the biomarker conditional vaccine efficacy adjusting for baseline covariates. We also propose a perturbation resampling method for making simultaneous inference on conditional vaccine efficacy over the values of the biomarker. We illustrate our method with datasets from two phase 3 dengue vaccine efficacy trials.