Objective:To establish and evaluate a second-tier screening method for neonatal congenital adrenal hyperplasia (CAH) and develop appropriate screening interpretation criteria.Methods:We employed liquid chromatography-tandem mass spectrometry to simultaneously detect five steroid hormones in dried blood spots: 17α-hydroxyprogesterone (17α-OHP), androstenedione (A4), 11-deoxycortisol (11-DOC), 21-deoxycortisol (21-DOC), and cortisol (F), calculating (17α-OHP+A4)/F and (17α-OHP+21-DOC)/F ratios for second-tier CAH screening. The study utilized 429 residual dried blood spot samples from neonates (0-7 days) who completed first-tier screening between January 2020 and March 2024 in Guangzhou Women and Children's Medical Center, Guangzhou Medical University, including first-tier negatives ( n=369), confirmed false positives ( n=50), and CYP21A2-confirmed 21-hydroxylase deficiency patients ( n=10). Mann-Whitney U and Kruskal-Wallis tests analyzed steroid concentration variations across gestational ages and birth weights in all negative samples, with reference intervals established via P2.5- P97.5 percentiles and screening cutoffs set at population P97.5. Receiver operating characteristic (ROC) curve analysis identified optimal interpretation indicators among steroid hormone profiles, with second-tier screening performance evaluated by comparing sensitivity and specificity across different steroid hormone indicators to establish the optimal diagnostic criteria. Results:The five steroid hormones demonstrated intra-assay precision with coefficient of variation (CV) of 9.8%-14.2% and inter-assay precision with CV of 4.7%-14.4% across three different concentration levels of quality control materials. Accuracy ranged from 98.5% to 110.0% and the lower limits of quantification were 0.25 ng/ml for 17α-OHP, 0.05 ng/ml for A4/11-DOC, 0.31 ng/ml for 21-DOC, and 0.1 ng/ml for F. Stratification by gestational age categorized 17α-OHP into ≤31, 32-34, and ≥35 weeks; A4 into ≤31, 32-36, and ≥37 weeks; and 11-DOC into ≤31 and ≥32 weeks, while the remaining indicators were not stratified. When grouped by birth weight (low/normal), all measured parameters except 21-DOC showed statistically significant differences between groups (all P<0.05). Established reference intervals included 17α-OHP: 0.53-7.82 ng/ml (≤31 weeks), <0.25-3.60 ng/ml (32-34 weeks), <0.25-1.64 ng/ml (≥35 weeks); A4: 0.12-2.36 ng/ml (≤31 weeks), <0.05-1.45 ng/ml (32-36 weeks), 0.17-0.95 ng/ml (≥37 weeks); 11-DOC: 0.43-4.04 ng/ml (≤31 weeks), 0.08-1.46 ng/ml (≥32 weeks); F: 1.70-83.70 ng/ml; 21-DOC: <0.31-0.69 ng/ml; (17α-OHP+A4)/F: 0.01-0.74; and (17α-OHP+21-DOC)/F: 0.01-0.69. Comprehensive comparison of CAH second-tier screening performance demonstrated that interpretation based on elevated 17α-OHP accompanied by either elevated 21-DOC or elevated ratios [(17α-OHP+A4)/F or (17α-OHP+21-DOC)/F] achieved 100% sensitivity, 96% specificity, and a 96% reduction in false-positive rate. Conclusion:The application of liquid chromatography-tandem mass spectrometry for multi-steroid hormone profiling in second-tier neonatal CAH screening, utilizing gestational age-specific 17α-OHP cutoffs combined with elevated 21-DOC or ratio criteria, demonstrated 100% screening sensitivity while substantially reducing false-positive rates from primary screening, though further validation with expanded sample sizes remains necessary.

Objective:To establish and evaluate a second-tier screening method for neonatal congenital adrenal hyperplasia (CAH) and develop appropriate screening interpretation criteria.Methods:We employed liquid chromatography-tandem mass spectrometry to simultaneously detect five steroid hormones in dried blood spots: 17α-hydroxyprogesterone (17α-OHP), androstenedione (A4), 11-deoxycortisol (11-DOC), 21-deoxycortisol (21-DOC), and cortisol (F), calculating (17α-OHP+A4)/F and (17α-OHP+21-DOC)/F ratios for second-tier CAH screening. The study utilized 429 residual dried blood spot samples from neonates (0-7 days) who completed first-tier screening between January 2020 and March 2024 in Guangzhou Women and Children's Medical Center, Guangzhou Medical University, including first-tier negatives ( n=369), confirmed false positives ( n=50), and CYP21A2-confirmed 21-hydroxylase deficiency patients ( n=10). Mann-Whitney U and Kruskal-Wallis tests analyzed steroid concentration variations across gestational ages and birth weights in all negative samples, with reference intervals established via P2.5- P97.5 percentiles and screening cutoffs set at population P97.5. Receiver operating characteristic (ROC) curve analysis identified optimal interpretation indicators among steroid hormone profiles, with second-tier screening performance evaluated by comparing sensitivity and specificity across different steroid hormone indicators to establish the optimal diagnostic criteria. Results:The five steroid hormones demonstrated intra-assay precision with coefficient of variation (CV) of 9.8%-14.2% and inter-assay precision with CV of 4.7%-14.4% across three different concentration levels of quality control materials. Accuracy ranged from 98.5% to 110.0% and the lower limits of quantification were 0.25 ng/ml for 17α-OHP, 0.05 ng/ml for A4/11-DOC, 0.31 ng/ml for 21-DOC, and 0.1 ng/ml for F. Stratification by gestational age categorized 17α-OHP into ≤31, 32-34, and ≥35 weeks; A4 into ≤31, 32-36, and ≥37 weeks; and 11-DOC into ≤31 and ≥32 weeks, while the remaining indicators were not stratified. When grouped by birth weight (low/normal), all measured parameters except 21-DOC showed statistically significant differences between groups (all P<0.05). Established reference intervals included 17α-OHP: 0.53-7.82 ng/ml (≤31 weeks), <0.25-3.60 ng/ml (32-34 weeks), <0.25-1.64 ng/ml (≥35 weeks); A4: 0.12-2.36 ng/ml (≤31 weeks), <0.05-1.45 ng/ml (32-36 weeks), 0.17-0.95 ng/ml (≥37 weeks); 11-DOC: 0.43-4.04 ng/ml (≤31 weeks), 0.08-1.46 ng/ml (≥32 weeks); F: 1.70-83.70 ng/ml; 21-DOC: <0.31-0.69 ng/ml; (17α-OHP+A4)/F: 0.01-0.74; and (17α-OHP+21-DOC)/F: 0.01-0.69. Comprehensive comparison of CAH second-tier screening performance demonstrated that interpretation based on elevated 17α-OHP accompanied by either elevated 21-DOC or elevated ratios [(17α-OHP+A4)/F or (17α-OHP+21-DOC)/F] achieved 100% sensitivity, 96% specificity, and a 96% reduction in false-positive rate. Conclusion:The application of liquid chromatography-tandem mass spectrometry for multi-steroid hormone profiling in second-tier neonatal CAH screening, utilizing gestational age-specific 17α-OHP cutoffs combined with elevated 21-DOC or ratio criteria, demonstrated 100% screening sensitivity while substantially reducing false-positive rates from primary screening, though further validation with expanded sample sizes remains necessary.

Objective:To investigate the lipidomics differences of CD4+T immune cells in diabetic kidney disease(DKD)mice,and screen out the differential metabolites with biological significance.Methods:CD4(L3T4)MicroBeads immunomagnetic beads were used to isolate CD4+T immune cells from spleen of BKS.Cg-Dock7m+/+Leprdb/J mice with spontaneous DKD;the purity of CD4+T cells were identified by flow cytometry.The non-targeted lipidomics of CD4+T cells were detected by LC-MS/MS,and the differ-ential metabolites were analyzed.Results:A total of 463 metabolites were detected by LC-MS.PCA and OPLS-DA analysis showed that the metabolic components were significantly separated;twenty-four differential metabolites were screened out.KEGG and enrich-ment analysis showed that the differential metabolites involved in the disorder of glycerol phospholipid metabolism.Conclusion:Phos-pholipid metabolism of CD4+T cells is closely related to the occurrence of DKD.Phospholipid metabolism targeting DKD CD4+T cells in DKD may be a new direction of DKD treatment.

Objective: To investigate the longitudinal association of plasma Irisin concentrations with changes in blood pressure (BP) levels among children,and to assess the moderating effect of physical activity (PA) or sedentary behavior (SB) on the relationship between Irisin levels and BP. Methods: Based on a cohort study, a cluster sampling method was used to select 3 651 school aged children from five schools in Guangzhou in 2017 at the baseline survey and follow up in 2019. Both at baseline and during follow up, PA and SB were assessed by validated questionnaires, and BP levels were measured by an electronic sphygmomanometer. A final sample of 521 children were enrolled based on the PA and SB at baseline. Plasma Irisin concentrations were measured by ELISA at baseline. Logistic regression analysis was recruited for exploring the associations of plasma Irisin concentrations with changes in BP. Moderating effects of PA and SB on the relationship between Irisin concentrations and BP were estimated using stratified analysis. Results: Logistic regression analysis indicated that there was no significant association between Irisin concentrations and changes in BP levels among children ( OR =0.98, P >0.05). After stratification for SB, Irisin levels in the low SB subgroup were inversely associated with changes in diastolic blood pressure ( OR=0.87, 95%CI=0.77-0.98, P =0.02). In addition, SB level had a moderating effect on the relationship between Irisin levels and the DBP changes ( P =0.01). Conclusion Increased Irisin concentration is associated with the decrease of DBP level among low SB children. Furthermore, SB level shows moderating role in the relationship between Irisin concentrations and changes in DBP levels.

Humans ; Autistic Disorder ; China/epidemiology* ; Cohort Studies ; Autism Spectrum Disorder/genetics* ; Asian People

Objective: To explore the longitudinal association of the levels of plasma irisin among children with changes in obesity related parameters and newly onset obesity, and to explore whether physical activity (PA) and sedentary behavior(SB) have regulatory effects, to provide a scientific basis for the prevention and control of childhood obesity work. Methods: Cluster random sampling method was used to select 521 children from five schools in Guangzhou in 2017 at baseline and were followed up in 2019. A based on baseline PA and SB, children who meet the following criterion were selected:moderate vigorous intensity PA≥60 min/d or <150 min/week; and gender , age specific SB≥ P 75 or SB < P 25 . Plasma irisin concentration was measured in all the selected children. Multiple linear regression and Logistic regression were conducted to analyze the association. Results: The two year cumulative incidence of obesity, overweight and obesity, and central obesity was 2.82%, 6.57%, and 6.81%, respectively. There was no statistically significant association between plasma irisin levels and changes in obesity related parameters, newly onset overweight obesity or central obesity among children ( P >0.05). After stratified by PA, the irisin concentration in the low PA group was positively associated with weight change ( B=0.229, P =0.03). After stratified by SB, the irisin concentration in the low SB group was positively associated with the height change ( B=0.210, P <0.05). In addition, PA level and SB level both had a moderating effect on the association between plasma irisin levels and the weight change ( P PA=0.01, P SB =0.05). Conclusion PA and SB show moderating effect on plasma irisin concentration and weight gain. No association of irisin concentration with newly onset overweight or obesity among children has been found.

Objective: To determine imaging features that may help predict the presence of placenta accreta, placenta increta or placenta percreta on prenatal MRI scanning in order to identify the most diagnostic findings. Methods: The prenatal MRI scan data of placenta accreta, placenta increta or placenta percreta (placenta implantation group, n=15) and normal placenta(placenta normal group, n=15) diagnosed and treated by surgical pathology from January 2010 to December 2017 in the People's Hospital of Huadu District were retrospectively analyzed.Two expert MRI doctors were blinded to the patients' true diagnosis and were asked to score a total of 10 MRI features of the placenta and adjacent structures.The interrater reliability was assessed using kappa statistics.The features with a moderate kappa statistic or better(kappa>0.40 ) were then compared with the true diagnosis for each observer. Results: Eight of the scored features had an interobserver reliability of kappa>0.40: placenta previa(κ=0.89); abnormal uterine bulging(κ=0.57); intraplacental hemorrhage(κ=0.45); the presence of dark intraplacental bands on T₂W imaging(κ=0.76); flow-empty blood vessels in placenta(κ=0.67); border on placenta and uterus blurring(κ=0.63); heterogeneity of signal intensity on T₂-weighted(T₂W) imaging(κ=0.53); and continuity of myometrium was interrupted(κ=0.64). Using Fisher's two-sided exact test, there were statistically significant differences between the proportion of patients with placental invasion and those without placental invasion for three of the features: abnormal uterine bulging(P=0.015, P=0.011); heterogeneity of T₂W imaging signal intensity(P=0.006, P=0.013); and presence of dark intraplacental bands on T₂W imaging(P=0.032, P=0.010). Conclusion MRI can be a useful adjunct to ultrasound in diagnosing placenta accreta prenatally.Three features that are seen on MRI in patients with placental invasion appear to be useful for diagnosis: uterine bulging; heterogeneous signal intensity within the placenta; and the presence of dark intraplacental bands on T₂W imaging.

Objective: To investigate prospectively molecular and clinical characteristics of infants with congenital hypothyroidism (CH) caused by DUOX2 mutations in Guangzhou. Methods: A population-based cohort of 83 patients with CH were recruited based on newborn screening results among 108 899 newborns who were born in Guangzhou between April 1 and September 30 in 2015.Genetic analysis of DUOX2 hotspots(including 11 exons)by PCR-direct sequencing was performed in 52 patients with suspected thyroid dyshormonogenesis (SDH) according to thyroid ultrasound at diagnosis.All the patients were followed up for 3 years.The data of this cohort study(prevalence of CH, detection rate of DUOX2, clinical features) were compared with those of 96 patients with SDH in 2011-2012. Results: (1) The incidence of CH in 2015 was 1∶1 312, and 73.5%(61/83 cases) of CH patients were classified as SDH.Compared with those founded in 2011-2012, the incidence of CH was increased (1∶1 312 vs.1∶2 779), and the difference was significant (P<0.001), while the frequency of SDH was not different significantly (73.5%vs.76.6%, P=0.593). (2)There were 27 cases(51.9%) with SDH detected DUOX2 hotspots variants, including 6 cases with biallelic variants, 21 cases with monoallelic variants, and 1 possible new pathogenic variant p. S1091F.The p. K530X was the most common mutation accounting for 51.5%(17/33 cases) detected allelic and involving in 16 cases (30.8%) with SDH.Novel p. S1091F variant was probably damaging variant.Both detected rate and spectrum of DUOX2 variant were not significantly different compared with those in 2011-2012 (all P>0.05). (3) There were no significant differences in the levels of thyrotropin (bsTSH), serum TSH (sTSH), free thyroxine (FT4) and thyroglobulin in neonates with dry blood spot at diagnosis between children with DUOX2 and without DUOX2 variants cases(all P>0.05). Among 27 cases, 24(88.9%) patients with DUOX2 mutation were transient CH, and 3 cases were permanent CH. Conclusions The incidence of CH was increased in last few years in Guangzhou.Most of them were SDH, and 51.9% of SDH cases had DUOX2 hotspots variants.Temporary CH is the main clinical outcome.The p. K530X was the most common mutation in this cohort population.

Objective: To evaluate the response of neoadjuvant chemotherapy on osteosarcoma by semi-quantitative parameters of dynamic contrast-enhancement magnetic resonance imaging (DCE-MRI). Methods: Retrospectively analysis 25 cases of osteosarcoma confirmed by pathology.All cases received DCE-MRI scan before and after 4 cycles of neoadjuvant chemotherapy.The following semi-quantitative parameters were calculated by post-processing software: early dynamic enhancement wash-in slope (Slope), maximum signal intensity (SI_max), time to peak (TTP), signal enhanced extent (SEE), peak percent enhancement (PPE), wash out rate (WOR), enhancement rate (R). All cases were divided into good response group (tumor necrosis rate ≥90%, n=12) and non-response group (tumor necrosis rate <90%, n=13) according to the Huvos grading method. Semi-quantitative parameters of DCE-MRI before and after neoadjuvant chemotherapy between good response group and non-response group were compared by Mann-whitney U test. Receiver operating characteristic curve (ROC) analysis was performed to evaluate the efficacy of different good response group and non-response group after neoadjuvant chemotherapy (NAC). Results: Slope, SI_max, TTP, SEE, PPE, WOR, R, TTP, WOR before and after NAC in good response group were significant different (P<0. 05), but only SI_max, SEE in non-response group. TTP, R were significant different between the above two groups before NAC, and Slope, SI_max, TTP, SEE, WOR, R were significant different after NAC (P<0.05). ROC was used to predict the diagnostic efficiency of various parameters for tumor necrosis rate after osteosarcoma NAC. It was found that the sensitivity and specificity of Slope, TTP and R parameters for predicting the response of osteosarcoma after chemotherapy were 83.3% and 92.3%, 91.7% and 69.2%, 84.6% and 75.0% respectively. The area under the curve (AUC) were 0.872 (95% CI: 0.716 to 1.027), 0.846 (95% CI: 0.685 to 1.007), 0.833 (95% CI: 0.662 to 1.004), the cut-off value were 0.032, 175 s, 5.441, Youden index were 0.756, 0.609, 0.596, respectively. Conclusions Slope, TTP, R were the most valuable semi-quantitative parameter of DCE-MRI to predict the response of NAC in osteosarcoma.

Objective: To explore the TPO, DUOX2 and DUOXA2 genotypes and phenotypes of children with permanent congenital hypothyroidism(PCH) suspected dyshormonogenesis in Guangzhou, identified and treated at Guangzhou Newborn Screening Center. Six of them were born between 2011 and 2012. Method: Retrospectively analyzed the clinical data of 9 children with PCH suspected dyshormonogenesis. Genetic analysis of TPO, DUOX2 and DUOXA2 genes were performed with Sanger sequencing. Result: Of the 9 patients, four were identified variants in TPO gene including three cases with biallelic variants and one case with monoallelic variant. Novel c. 1784G>C( p. R595T) variant in TPO was predicted to be damaging by SIFT and PolyPhen-2. Four patients harbored monoallelic known variants in DUOX2 gene and the other one harbored heterozygous known mutation c. 738C>G(p.Y246X) in DUOXA2 gene.Two adolescent patients with biallelic variants in TPO gene showed classical PCH phenotypes with thyroid goiter or nodules. The six patients with monoallelic variant in TPO, DUOX2 or DUOXA2 presented variable phenotypes. Among the 433 578 newborns in the 2011-2012 cohort, there were 156 cases of CH. Six of these cases were PCH suspected dyshormonogenesis, among which 1 case was confirmed TPO biallelic variants and 5 cases were monoallelic variants of TPO, DUOX2, or DUOXA2 genes. Conclusion TPO and DUOX2 variants are the common molecular pathogenesis in children with PCH suspected dyshormonogenesis. Monoallelic variants in TPO, DUOX2 or DUOXA2 are associated with PCH and showed wide variability in their phenotypes. The novel variant p. R595T in TPO is probably a pathologic variant. The prevalence of PCH caused by TPO gene defects is rare in Guangzhou.