Development and validation of real-time recombinase polymerase amplification-based assays for detecting HPV16 and HPV18 DNA.

IMPORTANCE: HPV DNA screening is an effective approach for the prevention of cervical cancer. The novel real-time recombinase polymerase amplification-based HPV detection systems we developed constitute an improvement over the HPV detection methods currently used in clinical practice and should help to extend cervical cancer screening in the future, particularly in point-of-care test settings.

Skeletal Muscle Mass Measurement Using Cone-Beam Computed Tomography in Patients With Head and Neck Cancer.

Background and purpose: Head and neck cancer (HNC) patients usually present with malnutrition during radiotherapy, leading to loss of skeletal muscle mass (SMM) and poor clinical outcomes. CT has been used in clinical practice for measuring SMM in cancer patients. However, its clinical application for monitoring SMM is limited by the expensive price and high radiation exposure. This study aimed to investigate the feasibility of cone-beam computed tomography (CBCT) for assessing SMM and its changes in HNC patients undergoing radiotherapy. Materials and methods: This study was divided into two parts. In part 1 (n = 32), the cross-sectional of skeletal muscle area (SMA) at the third cervical vertebra (C3) based on CBCT and computed tomography (CT) was assessed. In part 2 (n = 30), CT and CBCT were performed, and patients' weight was measured before and at four different time points during radiotherapy. SMAs at C3 were independently identified by three senior radiation oncologists. The interobserver agreement of SMA on CBCT (SMACBCT) findings was analyzed using the intraclass correlation coefficient (ICC). One-way analysis of variance was used to evaluate the interobserver variability and statistical significance for SMA measurements. CBCT and CT measurement differences and correlations were analyzed using paired sample t-test and Pearson correlation analysis, respectively. The Krouwer variant of the Bland-Altman plot was used to analyze the agreement of SMA measurements between CBCT and CT. A simple linear regression model was used to analyze the relationship of SMA measurements between the two imaging techniques, and the equation was established. A repeated-measures ANOVA was performed to evaluate the effects and interactions between weight loss, SMA loss, and time. Results: SMACBCT demonstrated excellent interobserver reliability; no significant difference between SMACBCT and SMA on CT (SMACT) at C3 was observed in all patients. The SMACBCT and SMACT were highly correlated (r = 0.966; 95% confidence interval = 0.955-0.975; p < 0.001). Bland-Altman analysis revealed that SMACBCT was generally higher than SMACT. The predicted SMA value at C3 on CT using CBCT was similar to the actual value. Moreover, significant differences between SMA and weight loss (F =10.99, p = 0.002), groups (weight loss and SMA loss) and times (4 time points) (F = 3.93, p = 0.013), and mean percent loss over time (F = 7.618, p < 0.001) were noted. Conclusion: CBCT may be used as an alternative for CT to measure SMA in HNC patients during radiotherapy.

Genome-Wide Placental Gene Methylations in Gestational Diabetes Mellitus, Fetal Growth and Metabolic Health Biomarkers in Cord Blood.

Gestational diabetes mellitus (GDM) "program" an elevated risk of metabolic syndrome in the offspring. Epigenetic alterations are a suspected mechanism. GDM has been associated with placental DNA methylation changes in some epigenome-wide association studies. It remains unclear which genes or pathways are affected, and whether any placental differential gene methylations are correlated to fetal growth or circulating metabolic health biomarkers. In an epigenome-wide association study using the Infinium MethylationEPIC Beadchip, we sought to identify genome-wide placental differentially methylated genes and enriched pathways in GDM, and to assess the correlations with fetal growth and metabolic health biomarkers in cord blood. The study samples were 30 pairs of term placentas in GDM vs. euglycemic pregnancies (controls) matched by infant sex and gestational age at delivery in the Shanghai Birth Cohort. Cord blood metabolic health biomarkers included insulin, C-peptide, proinsulin, IGF-I, IGF-II, leptin and adiponectin. Adjusting for maternal age, pre-pregnancy BMI, parity, mode of delivery and placental cell type heterogeneity, 256 differentially methylated positions (DMPs,130 hypermethylated and 126 hypomethylated) were detected between GDM and control groups accounting for multiple tests with false discovery rate <0.05 and beta-value difference >0.05. WSCD2 was identified as a differentially methylated gene in both site- and region-level analyses. We validated 7 hypermethylated (CYP1A2, GFRA1, HDAC4, LIMS2, NAV3, PAX6, UPK1B) and 10 hypomethylated (DPP10, CPLX1, CSMD2, GPR133, NRXN1, PCSK9, PENK, PRDM16, PTPRN2, TNXB) genes reported in previous epigenome-wide association studies. We did not find any enriched pathway accounting for multiple tests. DMPs in 11 genes (CYP2D7P1, PCDHB15, ERG, SIRPB1, DKK2, RAPGEF5, CACNA2D4, PCSK9, TSNARE1, CADM2, KCNAB2) were correlated with birth weight (z score) accounting for multiple tests. There were no significant correlations between placental gene methylations and cord blood biomarkers. In conclusions, GDM was associated with DNA methylation changes in a number of placental genes, but these placental gene methylations were uncorrelated to the observed metabolic health biomarkers (fetal growth factors, leptin and adiponectin) in cord blood. We validated 17 differentially methylated placental genes in GDM, and identified 11 differentially methylated genes relevant to fetal growth.

Deep Learning Prediction of Ovarian Malignancy at US Compared with O-RADS and Expert Assessment.

Background Deep learning (DL) algorithms could improve the classification of ovarian tumors assessed with multimodal US. Purpose To develop DL algorithms for the automated classification of benign versus malignant ovarian tumors assessed with US and to compare algorithm performance to Ovarian-Adnexal Reporting and Data System (O-RADS) and subjective expert assessment for malignancy. Materials and Methods This retrospective study included consecutive women with ovarian tumors undergoing gray scale and color Doppler US from January 2019 to November 2019. Histopathologic analysis was the reference standard. The data set was divided into training (70%), validation (10%), and test (20%) sets. Algorithms modified from residual network (ResNet) with two fusion strategies (feature fusion [hereafter, DLfeature] or decision fusion [hereafter, DLdecision]) were developed. DL prediction of malignancy was compared with O-RADS risk categorization and expert assessment by area under the receiver operating characteristic curve (AUC) analysis in the test set. Results A total of 422 women (mean age, 46.4 years ± 14.8 [SD]) with 304 benign and 118 malignant tumors were included; there were 337 women in the training and validation data set and 85 women in the test data set. DLfeature had an AUC of 0.93 (95% CI: 0.85, 0.97) for classifying malignant from benign ovarian tumors, comparable with O-RADS (AUC, 0.92; 95% CI: 0.85, 0.97; P = .88) and expert assessment (AUC, 0.97; 95% CI: 0.91, 0.99; P = .07), and similar to DLdecision (AUC, 0.90; 95% CI: 0.82, 0.96; P = .29). DLdecision, DLfeature, O-RADS, and expert assessment achieved sensitivities of 92%, 92%, 92%, and 96%, respectively, and specificities of 80%, 85%, 89%, and 87%, respectively, for malignancy. Conclusion Deep learning algorithms developed by using multimodal US images may distinguish malignant from benign ovarian tumors with diagnostic performance comparable to expert subjective and Ovarian-Adnexal Reporting and Data System assessment. © RSNA, 2022 Online supplemental material is available for this article.

Comparison of the Diagnostic Performances of Ultrasound-Based Models for Predicting Malignancy in Patients With Adnexal Masses.

AIM: This study aimed to compare different ultrasound-based International Ovarian Tumor Analysis (IOTA) prediction models, namely, the Simple Rules (SRs) the Assessment of Different NEoplasias in the adneXa (ADNEX) models, and the Risk of Malignancy Index (RMI), for the pre-operative diagnosis of adnexal mass. METHODS: This single-centre diagnostic accuracy study involved 486 patients. All ultrasound examinations were analyzed and the prediction models were applied. Pathology was the clinical reference standard. The diagnostic performances of prediction models were measured by evaluating receiver-operating characteristic curves, sensitivities, specificities, positive and negative predictive values, positive and negative likelihood ratios, and diagnostic odds ratios. RESULTS: To discriminate benign and malignant tumors, areas under the ROC curves (AUCs) for ADNEX models were 0.94 (95% CI: 0.92-0.96) with CA125 and 0.94 (95% CI: 0.91-0.96) without CA125, which were significantly higher than the AUCs for RMI I-III: 0.87 (95% CI: 0.83-0.90), 0.83 (95% CI: 0.80-0.86), and 0.82 (95% CI: 0.78-0.86), (all P < 0.0001). At a cut-off of 10%, the ADNEX model with CA125 had the highest sensitivity (0.93; 95% CI: 0.87-0.97) compared with the other models. The SRs model achieved a sensitivity of 0.93 (95% CI: 0.86-0.97) and a specificity of 0.86 (95% CI: 0.82-0.89) when inconclusive diagnoses (11.7%) were classified as malignant. CONCLUSION: ADNEX and SRs models were excellent at characterising adnexal masses which were superior to the RMI in Chinese patients.

Pregnancy Outcomes in Patients With Adult-Onset Still's Disease: A Cohort Study From China.

Objective: Adult-onset Still's disease (AOSD) is an autoinflammatory disease with a higher prevalence rate in young females. The purpose of this study is to investigate whether AOSD has an adverse impact on pregnancy outcomes, or conversely exacerbated by pregnancy. Methods: The outcomes of 191 pregnancies were evaluated in 86 female patients with AOSD. The generalized linear mixed model and propensity score matching method were conducted to evaluate the influence of AOSD on pregnancy outcomes. A dependent sample sign test was applied to assess the impact of pregnancy on the relapse of AOSD. Results: The results showed that the post-AOSD group had a lower proportion of normal delivery (25.0 vs. 52.4%, p = 0.036) and a higher proportion of spontaneous abortion (STA) (18.8 vs. 0.6%, p = 0.002) compared with the pre-AOSD group. Moreover, pregnancy after being diagnosed with AOSD was a significant high risk factor of STA (adjusted OR = 4.577, 95% CI: 4.166-845.119; p = 0.003). Disease flare upon conception was observed in one of 16 post-AOSD pregnancies (p = 1.000). There were 11 patients with new-onset AOSD during gestation or postpartum, among which five (45.4%) evolved into the polycyclic course. Conclusions: AOSD patients might suffer from a higher risk of STA, however, pregnancy might not be related with the exacerbation of diagnosed AOSD. New-onset AOSD during gestation or postpartum tend to evolve into the polycyclic course.

Sex dimorphism in the associations of gestational diabetes with cord blood adiponectin and retinol-binding protein 4.

INTRODUCTION: Gestational diabetes (GD) is associated with impaired insulin sensitivity in newborns. Adiponectin and retinol-binding protein 4 (RBP-4) are involved in regulating insulin sensitivity. Females are more likely to develop diabetes at young ages than males. We tested the hypothesis that GD may affect RBP-4 and adiponectin levels in early life, and there may be sex-dimorphic associations. RESEARCH DESIGN AND METHODS: In a nested case-control study of 153 matched pairs of neonates of mothers with GD and euglycemic pregnancies in the Shanghai Birth Cohort, we evaluated cord plasma leptin, high molecular weight (HMW) and total adiponectin and RBP-4 concentrations. RESULTS: Comparing GD versus euglycemic pregnancies adjusted for maternal and neonatal characteristics in female newborns, cord plasma total adiponectin (mean±SD: 30.8±14.3 vs 37.1±16.1 µg/mL, p=0.048) and HMW adiponectin (14.6±7.7 vs 19.3±8.3 µg/mL, p=0.004) concentrations were lower, while RBP-4 concentrations were higher (21.7±5.4 vs 20.0±4.8 µg/mL, p=0.007). In contrast, there were no differences in male newborns (all p>0.2). RBP-4 concentrations were higher in female versus male newborns (21.7±5.4 vs 18.8±4.5 µg/mL, p<0.001) in GD pregnancies only. HMW adiponectin concentrations were significantly higher in female versus male newborns in euglycemic pregnancies only (19.3±8.3 vs 16.1±7.4 µg/mL, p=0.014). CONCLUSIONS: GD was associated with lower cord plasma HMW adiponectin and higher RBP-4 concentrations in female newborns only. The study is the first to reveal a sex-dimorphic early life impact of GD on metabolic health biomarkers in the offspring. GD may alter the normal presence (HMW adiponectin) or absence (RBP-4) of sex dimorphism in some insulin sensitivity regulation-relevant adipokines in early life.

The Sex Specific Association Between Maternal Gestational Diabetes and Offspring Metabolic Status at 1 Year of Age.

Previous studies showed the association between maternal GDM and long-term effects of overweight in offspring. However, the nature of this association in the early postnatal period is still undetermined. The aim of this prospective cohort study was to evaluate whether maternal GDM is associated with overweight and obesity status in offspring at age 1 year. We studied 1167 infants born at a large obstetrical care hospital including 778 normal glucose tolerance (NGT) and 389 GDM pregnancies, matched in a 1:2 ratio according to offspring's gender, during the years 2016-2017. Overweight and obesity status in offspring of both groups were evaluated at 1 year of age through questionnaires. Infant outcomes were defined according to the WHO Child Growth Standards based on the length-based BMI-for-age. Female offspring from the GDM group exhibited a higher mean BMI (17.2 vs. 16.6, p < 0.01), a higher rate of obesity (13.9% vs. 7.7%; p < 0.05), and overweight (33.1% vs. 23.5%; p < 0.05) as compared to the NGT female group. In the multivariable regression model, maternal GDM was found to be independently and significantly associated with overweight or obesity in 1-year aged female offspring only (OR 1.61, 95% CI 1.09-2.37, p < 0.05). We found a sex specific association between maternal GDM and the overweight risk only in female offspring at 1 year of age.

Vitamin A and E Nutritional Status in Relation to Leptin, Adiponectin, IGF-I and IGF-II in Early Life - a Birth Cohort Study.

The metabolic health effects of vitamin A and E nutritional status in early life are largely unknown. We assessed whether vitamin A and vitamin E nutritional status may affect circulating leptin, adiponectin, insulin-like growth factor (IGF)-I and IGF-II levels in early life in humans. In a singleton birth cohort (n = 248), vitamin A and E nutritional status in fetuses/newborns were assessed by cord plasma concentrations of retinol, ß-carotene, α- and γ-tocopherols. The primary outcomes were cord plasma leptin, adiponectin, IGF-I and IGF-II concentrations. Cord plasma retinol was significantly positively correlated to IGF-I in girls (r = 0.42, P < 0.0001) but not in boys (r = 0.14, P = 0.11). Adjusting for maternal and newborn's characteristics, one log unit increase in cord plasma retinol was associated with a 28.0% (95% CI: 11.1-47.5%) increase in IGF-I in girls (P < 0.001) but not in boys (P = 0.75). One log unit increment in cord plasma α-tocopherol was associated with a 6.6% (0.4-12.3%) decrease in adiponectin (P = 0.04), while one log unit increment in cord plasma γ-tocopherol was associated with a 21.2% (4.7-34.8%) decrease in leptin (P = 0.01). There may be a sex-specific association between retinol and IGF-I, a negative association between α-tocopherol and adiponectin, and a negative association between γ-tocopherol and leptin in early life in humans.

Early and Late Postnatal Accelerated Growth Have Distinct Effects on Metabolic Health in Normal Birth Weight Infants.

Accelerated growth in postnatal life in low birth weight infants has been associated with insulin resistance and metabolic syndrome-related disorders in later life. Postnatal accelerated growth in also common in normal birth weight infants, but little is known about the impact on metabolic health. In a prospective cohort study of 203 term normal birth weight infants, we evaluated the impacts of accelerated (Δweight Z score > 0.5) or decelerated (Δweight ΔZ < -0.5) growth during early (0-3 months) and late (3-12 months) postnatal life on metabolic health indicators at age 1-year. The primary outcomes were homeostasis model assessment of insulin resistance (HOMA-IR), ß-cell function [homeostasis model assessment of ß-cell function (HOMA-ß)], and fasting plasma lipids. Adjusting for maternal, paternal, and infant characteristics, accelerated growth during the first 3 months of life was associated with a 41.6% (95% confidence interval 8.9-84.2%) increase in HOMA-ß, and a 8.3% (0.7-15.4%) decrease in fasting plasma total cholesterols, and was not associated with HOMA-IR in infants at age 1-year. Accelerated growth during 3-12 months was associated with a 30.9% (3.3-66.0%) increase in HOMA-IR and was not associated with HOMA-ß. Neither accelerated nor decelerated growth was associated with fasting plasma triglycerides, high-density lipoprotein or low-density lipoprotein cholesterol concentrations in infants at age 1-year. Accelerated growth during early postnatal life may be beneficial for ß-cell function, but during late postnatal life harmful for insulin sensitivity in normal birth weight infants.

Postnatal weight gain induced by overfeeding pups and maternal high-fat diet during the lactation period modulates glucose metabolism and the production of pancreatic and gastrointestinal peptides.

The impact of rapid weight gain on glucose metabolism during the early postnatal period remains unclear. We investigated the influence of rapid weight gain under different nutritional conditions on glucose metabolism, focusing on the production of pancreatic and gastric peptides. On postnatal day (PND) 2, C57BL/6N pups were divided into three groups: control (C) pups whose dams were fed a control diet (10%kcal fat) and nursed 10 pups each; maternal high-fat diet (HFD) pups whose dams were fed an HFD (45%kcal fat) and nursed 10 pups each; and overfeeding (OF) pups whose dams were fed the control diet and nursed 4 pups each. Data were collected on PND 7, 14 and 21. The body weight gains of the HFD and OF pups were 1.2 times higher than that of the C pups. On PND 14, the HFD pups had higher blood glucose levels, but there were no significant differences in serum insulin levels between the HFD and C pups. The OF pups had higher blood glucose and serum insulin levels than that of the C pups. Insulin resistance was found in the HFD and OF pups. On PND 14, the content of incretins in the jejunum was increased in the OF pups, and acyl ghrelin in the stomach was upregulated in the HFD and OF pups. These results suggest that neonatal weight gain induced by overfeeding pups and maternal high-fat diet during the early postnatal period modulates the insulin sensitivity and the production of pancreatic and gastrointestinal peptides.

A maternal mouse diet with moderately high-fat levels does not lead to maternal obesity but causes mesenteric adipose tissue dysfunction in male offspring.

The impact of an increase in maternal fat consumption on fetal metabolic programming separately from maternal obesity remains unclear. The purpose of this study was to document the effect of in utero high-fat diet exposure on the development of metabolic syndrome characteristics in offspring. C57BL/6 female mice were fed either a control diet (10% fat) or a moderately high-fat (MHF) diet (45% fat) until delivery. All pups were fostered to mothers fed with the control diet. Pups were raised on the control diet and assessed until 35 weeks of age. The caloric intake from fat was significantly increased in the MHF dams compared with the control dams. There were no significant differences in the maternal weight at mating or at gestational Day 18 between the two groups. The MHF offspring did not become obese, but they developed hypertension and glucose intolerance. Moreover, the MHF offspring had significantly higher serum non-esterified fatty acid and triglyceride levels during the refeeding state following fasting as compared with the control offspring. Serum adiponectin levels were significantly lower, and the cell size of the mesenteric adipose tissue was significantly larger in the MHF offspring than in the control offspring. The mRNA levels of the proinflammatory macrophage markers in the mesenteric adipose tissue were significantly higher in the MHF offspring than those of the control offspring. These results suggest that in utero high-fat diet exposure causes hypertension and glucose intolerance resulting from mesenteric adipose tissue dysfunction in offspring, independently of maternal obesity.