The Application of the First Year Inventory for ASD Screening in China.

PURPOSE: The First Year Inventory (FYI) is a parent-report instrument, and is developed to assess behaviors of 12-month-old infants that could suggest risk for an eventual diagnosis of autism. This study was designed to examine the application of the FYI in the Chinese community. DESIGN AND METHODS: FYIs were completed at a community health center by 541 families during the child's physical examination at 12 months of age from 2013 to 2015. The weighted risk scores used in this study were based on US norms, and compared the FYI differences between China and the U.S. RESULTS: The total risk scores ranged from 5 to 42 points; the 95th percentile cutoff was 27.00(9.8 points higher than the 95th percentile cutoff in the US), the 98th percentile cutoff was 29.66(7.04 points higher than the 98th percentile cutoff in the US), and the 99th percentile cutoff was 31.83. Higher risk scores were found for boys than girls. Mothers with a junior college education reported significantly higher FYI risk scores than other three groups including high school, college graduates and post-graduates. CONCLUSIONS: There were no significant effects of birth parity, investigator, or investigation year on risk scores. Large-scale longitudinal research is encouraged in the future to develop an early detection model of autism in China.

Circular RNA expression profile and potential function of hsa_circ_0045272 in systemic lupus erythematosus.

Circular RNAs (circRNAs) represent a class of non-coding RNAs that form covalently closed RNA circles with extensive expression and conservation in mammals. Circular RNAs regulate gene expression through acting as competitive endogenous RNAs (ceRNAs) and modulating gene transcription. Accumulating evidence supports the implication of circRNAs in a variety of human diseases, but studies of circRNA role in systemic lupus erythematosus (SLE) are lacking. The present study measured the circRNA expression profiles in T cells from patients with SLE and healthy controls with human circRNA microarray and identified 127 differentially expressed circRNAs in SLE patients. Down-regulation of hsa_circ_0045272 in SLE T cells was verified with quantitative PCR. Jurkat cells with stable hsa_circ_0045272 knockdown were generated using specific lentiviral short hairpin RNA for functional studies. Flow cytometric analysis indicated that hsa_circ_0045272 knockdown significantly up-regulated the early apoptosis of Jurkat cells. Meanwhile, ELISA showed that hsa_circ_0045272 knockdown significantly enhanced interleukin-2 production of activated Jurkat cells. Then, ceRNAs were predicted for hsa_circ_0045272 and the significant down-regulation of two mRNAs predicted as ceRNAs, NM_003466 (PAX8) and NM_015177 (DTX4), but not their corresponding proteins, was validated. Furthermore, dual luciferase reporter assay indicated binding of hsa_circ_0045272 with hsa-miR-6127. Circular RNA-mRNA co-expression networks showed the correlation of circRNAs with mRNAs and provided additional clues to circRNA functions. Our study demonstrated dysregulated circRNAs in SLE and revealed the function of hsa_circ_0045272 in negatively regulating apoptosis and interleukin-2 secretion and its potential mechanism. The implication of hsa_circ_0045272 and other abnormal circRNAs in SLE merits further investigation.

Characterization of the phytochelatins and their derivatives in rice exposed to cadmium based on high-performance liquid chromatography coupled with data-dependent hybrid linear ion trap orbitrap mass spectrometry.

RATIONALE: The identification and quantification of phytochelatins (PCs) and their derivatives are important to understand their roles in plant growth and development. A method couplling high-performance liquid chromatography with hybrid linear ion trap Orbitrap mass spectrometry (HPLC-LTQ/Orbitrap) was developed to screen PCs that have the same characteristic product ions. This approach was used for the fragmentation pattern analysis of glutathione (GSH) and PC standards, which allowed identification of the fragmentation pathways of their derivatives isolated from rice roots, stems and leaves. METHODS: In this study, we developed a method to detect and identify PCs and their derivatives in rice based on HPLC/LTQ-Orbitrap. Spectrum interpretation and MS/MS fragmentation patterns of PCs provide sufficient information to discover the novel PC derivatives. This approach includes precursor ion scan and product ion scan to detect and character the novel PC derivatives. RESULTS: Based on HCD-MS/MS fragmentation patterns, four PCs and 18 PC derivatives were identified. Among them, seven PC derivatives, i.e., iso-PC2 (Asn), iso-PC3 (Asn), iso-PC2 (Cys), des-γGlu-iso-PC3 (Ser), des-Cys-iso-PC2 (Glu), des-Cys-iso-PC3 (Glu) and des-Cys-iso-PC4 (Glu), have not been previously reported. This method was validated by profiling GSH, PCs and PC derivatives in rice. Preliminary results revealed that PCs and their derivatives, except GSH, are markedly induced by Cd treatment. CONCLUSIONS: The HPLC/LTQ-Orbitrap method was successfully developed for the identification of PCs and their derivatives. The C-terminal linked to Gly is replaced with Glu, Ser, Asn, Gln or Cys, thereby creating a family of chemicals that share several structural properties. This technique could be particularly useful for investigators studying plant metabolomics. Copyright © 2016 John Wiley & Sons, Ltd.

Associations between maternal lipid profile and pregnancy complications and perinatal outcomes: a population-based study from China.

BACKGROUND: Dyslipidemia in pregnancy are associated with gestational diabetes mellitus (GDM), preeclampsia, preterm birth and other adverse outcomes, which has been extensively studied in western countries. However, similar studies have rarely been conducted in Asian countries. Our study was aimed at investigating the associations between maternal dyslipidemia and adverse pregnancy outcomes among Chinese population. METHODS: Data were derived from 934 pairs of non-diabetic mothers and neonates between 2010 and 2011. Serum blood samples were assayed for fasting total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), and low-density lipoprotein-cholesterol (LDL-C) concentrations during the first, second and third trimesters. The present study explored the associations between maternal lipid profile and pregnancy complications and perinatal outcomes. The pregnancy complications included GDM, preeclampsia and intrahepatic cholestasis of pregnancy (ICP); the perinatal outcomes included preterm birth, small/large for gestational age (SGA/LGA) infants and macrosomia. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated and adjusted via stepwise logistic regression analysis. Optimal cut-off points were determined by ROC curve analysis. RESULTS: After adjustments for confounders, every unit elevation in third-trimester TG concentration was associated with increased risk for GDM (OR = 1.37, 95% CI: 1.18-1.58), preeclampsia (OR = 1.50, 95% CI: 1.16-1.93), ICP (OR = 1.28, 95% CI: 1.09-1.51), LGA (OR = 1.13, 95% CI: 1.02-1.26), macrosomia (OR = 1.19, 95% CI: 1.02-1.39) and decreased risk for SGA (OR = 0.63, 95% CI: 0.40-0.99); every unit increase in HDL-C concentration was associated with decreased risk for GDM and macrosomia, especially during the second trimester (GDM: OR = 0.10, 95% CI: 0.03-0.31; macrosomia: OR = 0.25, 95% CI: 0.09-0.73). The optimal cut-off points for third-trimester TG predicting GDM, preeclampsia, ICP, LGA and SGA were separately ≥ 3.871, 3.528, 3.177, 3.534 and ≤ 2.530 mmol/L. The optimal cut-off points for third-trimester HDL-C identifying GDM, macrosomia and SGA were respectively ≤ 1.712, 1.817 and ≥ 2.238 mmol/L. CONCLUSIONS: Among Chinese population, maternal high TG in late pregnancy was independently associated with increased risk of GDM, preeclampsia, ICP, LGA, macrosomia and decreased risk of SGA. Relative low maternal HDL-C during pregnancy was significantly associated with increased risk of GDM and macrosomia; whereas relative high HDL-C was a protective factor for both of them.

THE RELATIONSHIP BETWEEN SERUM 25-HYDROXYVITAMIN D AND GLUCOSE HOMEOSTASIS IN OBESE CHILDREN AND ADOLESCENTS IN ZHEJIANG, CHINA.

OBJECTIVE: Evidence of the association between vitamin D, insulin resistance, and oral disposition index (oDI) in obese children and adolescents is limited. To fill this research gap, we measured serum 25-hydroxyvitamin D (25[OH]D) levels in obese children and analyzed the relationship between serum 25(OH)D levels and glucose homeostasis. METHODS: Altogether, 348 obese and 445 nonobese children and adolescents (age, 6 to 16 years) were enrolled in this study. Obese children were divided into 4 subgroups: normal glucose tolerance (NGT), impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and combined IFG and IGT (IFG+IGT) according to oral glucose tolerance test results. We measured serum 25(OH)D levels and calculated the homeostasis model assessment (HOMA) of insulin resistance (IR), the whole-body insulin sensitivity index (WBISI), and the disposition index. RESULTS: The levels of 25(OH)D in the obese group were significantly lower than in the nonobese group; serum 25(OH)D level in the NGT subgroup was higher than those of the other 3 subgroups, and it was significantly inversely correlated with logHOMA-IR (r = -0.090; P = .045) and positively correlated with logWBISI and logHOMA-oDI (r = 0.091, P = .049; and r = 0.108, P = .046, respectively). Obese patients with vitamin D deficiency thus have a significantly higher risk of disturbances in glucose metabolism. CONCLUSION: 25(OH)D deficiency or insufficiency is quite common in obese children and adolescents in Zhejiang, China. Obese patients with 25(OH)D deficiency (<30 nmol/L) are shown to be at higher risk for abnormal glucose metabolism.

Elucidation of in vitro phase I metabolites of droperidol using UPLC-QTOF MS.

Droperidol, an antidopaminergic drug clinically used as an antiemetic and antipsychotic, has been reported to induce cardiac toxicity in patients. Due to the close relationship between drug metabolism and efficiency and toxicity, the present study aims to investigate the phase I metabolites using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The NADPH-supplemented phase I incubation system was used to elucidate the in vitro phase I metabolites. Five metabolites were detected after droperidol was incubated with phase I incubation mixture, including one hydrogenated droperidol, three oxidative metabolites, and one N-dealkylated droperidol, elucidated by individual retention time and MS/MS fragmentation. Due to the existed phase II metabolic reaction, further phase II metabolism should be investigated in the future. In conclusion, the phase I metabolism of droperidol was investigated in the present study, and five new metabolites were identified. The efficiency and toxicity of these phase I metabolites should be investigated in the future.

Construction of a lentiviral T/A vector for direct analysis of PCR-amplified promoters.

The promoter plays an important role in the regulation of gene expression. To analyze a promoter's activity, we developed a novel lentiviral T/A vector that contains two reporter genes, a luciferase (Luc2) gene and a green fluorescent protein (Venus) gene, that are linked via an internal ribosome entry site (IRES2). To test the performance of this vector, phosphoglycerate kinase-1 (PGK) and elongation factor-1α (EF1α) promoters were amplified by PCR and inserted into this lentiviral T/A vector using T4 DNA ligase, yielding two promoter-reporter vectors: pLent-T-PGK and pLent-T-EF1α. When these vectors were transfected into 293T cells, we observed a higher level of Venus expression under a fluorescence microscopy in the case of pLent-T-EF1α as compared to pLent-T-PGK. The results of the luciferase reporter assay showed that the ratio of the promoter activities of EF1α and PGK was approximately 9:1. The two promoter-reporter vectors were also packaged as lentiviral particles to conduct promoter activity assay in cultured cells. The ratio of the promoter activities of EF1α and PGK was 4.23:1 when they were infected into 293T cells at a multiplicity of infection of 1. This value is comparable to that of a parallel experiment using the commercial luciferase reporter vector pGL4.10 with an activity ratio of 5.99:1 for EF1α and PGK. These results indicate that lentiviral T/A vector will be a useful tool for analysis of promoter activity and specificity.

[Methylation analysis of CpG island DNA of FMR1 gene in the fragile X syndrome].

OBJECTIVE: To establish a method of methylation-sensitive restriction enzymes based quantitative PCR (MSRE-qPCR) for analysis of CpG island DNA of FMR1 gene, and to assess its value for molecular diagnosis of fragile X syndrome. METHODS: Thirty boys with mental retardation and abnormal repeats of 5'(CGG)n in the FMR1 gene and 20 mothers were analyzed by conventional PCR screening. Eag I was used to digest genomic DNA, and qPCR was performed to amplify CpG island in the FMR1 gene using both undigested and digested templates. Raw Ct values were obtained through quantitative PCR amplification. The degree of CpG island methylation was calculated by 2 - U+0394 U+0394 Ct. The result of MSRE-qPCR was verified by Southern blotting. 30 healthy females and 30 healthy males were used as controls to optimize the established MSRE-qPCR method. RESULTS: The ranges of 2 - U+0394 U+0394 Ct value for normal methylation, partial methylation and full methylation were determined. Among the 30 patients, 3 were found to have partial methylation of CpG island of the FMR1 gene, and 27 were found to have full methylation (3/30 results were verified by Southern blotting). Only 7 mothers were found abnormal methylation of CpG island of FMR1 gene, whilst the remaining 13 mothers were normal. CONCLUSION: MSRE-qPCR is a quick and reliable method for quantitative analysis of CpG island methylation status in FMR1 gene, which may provide a new strategy for the diagnosis of fragile X syndrome.

[Construction of Saccharomyces cerevisiae whole-cell biocatalyst system for conversion miltiradiene].

Tanshinones are the bioactive components of the Chinese medicinal herb Salvia miltiorrhiza, while its biosynthetic pathway remains to be characterized. Rapid identification and characterization of the genes correlated to tanshinones biosynthesis is very important. As one of the intermediates of tanshinones biosynthesis, the ferruginol content is relative low in both root and engineered bacteria. It is urgent to construct an efficient system for conversion of miltiradiene to ferruginol to obtain large amount of ferruginol as the substrates for further identifying other downstream genes involved in tanshinones biosynthesis. In this study, we constructed the whole-cell yeast biocatalysts co-expressing miltiradiene oxidase CYP76AH1 and cytochrome P450 reductases (SmCPR1) from Salvia miltiorrhiza, and then characterized it with RT-PCR. After permeabilization, the yeast whole-cell could catalyze turnover of miltiradiene to ferruginol efficiently through single-step biotransformation with a conversion efficiency up to 69.9%. The yeast whole-cell biocatalyst described here not only provide an efficient platform for producing ferruginol in recombinant yeast but also an alternative strategy for identifying other CYP genes involved in tanshinones biosynthesis.

Associations between cord blood metabolic factors and early-childhood growth and overweight and obesity.

Objective: This prospective cohort study was aimed at investigating the associations between cord blood metabolic factors and early-childhood growth, further elucidating the relationships between cord blood metabolites and overweight and obesity in early life. Methods: A total of 2,267 pairs of mothers and offspring were recruited in our study. Cord blood plasma was assayed for triglycerides (TGs), total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), C-peptide, insulin, and glycosylated hemoglobin type A1C (HbA1c) levels. Data of anthropometric measurements were collected from offspring at birth, 6 months, 12 months, and 18 months. Multiple linear regression models were used to evaluate the correlations between cord blood metabolic factors and weight Z-scores, body mass index (BMI) Z-scores, and weight gains at the early stage of life. Forward stepwise logistic regression analyses were applied to explore the associations between cord blood metabolic factors and early-childhood overweight and obesity. Receiver operating characteristic (ROC) curve analyses were applied to determine the optimal cutoff points for cord blood metabolic factors in predicting early-childhood overweight and obesity. Results: After adjustments for covariates, cord blood TG concentrations and TG/TC ratios were negatively associated with weight Z-scores from birth to 18 months. Cord blood C-peptide and HbA1c levels were inversely associated with weight Z-scores at 6 months and 18 months. Cord blood TG concentrations and TG/TC ratios were negatively correlated with BMI Z-scores up to 18 months. Cord blood C-peptide levels and HbA1c levels were inversely correlated with BMI Z-scores at 18 months. Cord blood TG, TG/TC ratios, C-peptide, and HbA1c had negative correlations with weight gains from birth to 6 months, but the correlations attenuated as time went on. Increase in cord blood TG and HbA1c levels and TG/TC ratios were significantly associated with decreased risks of overweight and obesity at 6 months, 12 months, and 18 months. Conclusions: Cord blood metabolic factors were significantly associated with early-childhood growth patterns.

Neural pathway connectivity and discharge changes between M1 and STN in hemiparkinsonian rats.

Alterations of electrophysiological activities, such as changed spike firing rates, reshaping the firing patterns, and aberrant frequency oscillations between the subthalamic nucleus (STN) and the primary motor cortex (M1), are thought to contribute to motor impairment in Parkinson's disease (PD). However, the alterations of electrophysiological characteristics of STN and M1 in PD are still unclear, especially under specific treadmill movement. To examine the relationship between electrophysiological activity in the STN-M1 pathway, extracellular spike trains and local field potential (LFPs) of STN and M1 were simultaneously recorded during resting and movement in unilateral 6-hydroxydopamine (6-OHDA) lesioned rats. The results showed that the identified STN neurons and M1 neurons exhibited abnormal neuronal activity after dopamine loss. The dopamine depletion altered the LFP power in STN and M1 whatever in rest or movement states. Furthermore, the enhanced synchronization of LFP oscillations after dopamine loss was found in 12-35 Hz (beta frequencies) between the STN and M1 during rest and movement. In addition, STN neurons were phase-locked firing to M1 oscillations at 12-35 Hz during rest epochs in 6-OHDA lesioned rats. The dopamine depletion also impaired the anatomical connectivity between the M1 and STN by injecting anterograde neuroanatomical tracing virus into M1 in control and PD rats. Collectively, impairment of' electrophysiological activity and anatomical connectivity in the M1-STN pathway may be the basis for dysfunction of the cortico-basal ganglia circuit, correlating with motor symptoms of PD.

[Estimation of PMI using late postmortem phenomena in the basis of 49 cases].

OBJECTIVE: To discuss the influencing factors of using late postmortem phenomena to estimate PMI and to provide experience for an accurate estimation. METHODS: Forty-nine corpses of late postmortem were collected in Shaoxing City, Zhuji area from 2004 to 2011. The related factors were analyzed including season, scene, estimated PMI, exact PMI, cause of death and main factors effected PMI, etc. RESULTS: Of all 49 cases, 20 corpses were outdoor, 11 were indoor and 18 were in water. Thirty-seven cases were successful to estimate PMI and 12 cases were unsuccessful. The main factors affected PMI were infection, poisoning, human destruction and high-pressure electric shock, etc. CONCLUSION: In general, PMI can be correctly estimated by late postmortem phenomenon. When the cases included infection, poisoning and human destruction, we should estimate PMI with the comprehensive analysis.

Rapid and Visual Detection of Toxoplasma gondii in Blood Samples from Pet Cats and Dogs by Loop-Mediated Isothermal Amplification.

Background: Toxoplasma gondii is an obligate intracellular parasite that invades nearly all nucleated cells of a broad spectrum of vertebrate hosts, and which may cause serious disease in immunocompromised patients, as well as in the immunologically incompetent fetus. This study aimed to establish a loop-mediated isothermal amplification (LAMP) technique to rapidly detect T. gondii in the blood infection by targeting the 529 bp repeat element of T. gondii. Materials and Methods: A turbidity monitoring system, together with visual reagent, was used to test the amplification result of the LAMP assay. In addition, the specificity and sensitivity of the LAMP assay were measured. Results: The results suggest that the successfully established LAMP assay profile can detect the DNA of T. gondii at 67°C within 40 min. The limit of detection of the LAMP assay was 101 copies/µL. No cross reaction occurred with Plasmodium vivax, Toxocara cati, Clonorchis sinensi, Spirometra mansoni or Cryptosporidium parvum. We validated the developed LAMP assay by detecting T. gondii in DNA extracted from 353 blood samples collected from domestic cats and dogs. The percentages of positive results in detecting these blood samples by LAMP and conventional PCR were 5.38% and 2.83%, respectively. Conclusions: Our findings show that the developed LAMP assay offers higher analytical sensitivity than conventional PCR and good analytical specificity, minimizes aerosol contamination, and can be applied to on-site rapid detection of T. gondii.

A machine learning-based diagnostic model for children with autism spectrum disorders complicated with intellectual disability.

Background: Early detection of children with autism spectrum disorder (ASD) and comorbid intellectual disability (ID) can help in individualized intervention. Appropriate assessment and diagnostic tools are lacking in primary care. This study aims to explore the applicability of machine learning (ML) methods in diagnosing ASD comorbid ID compared with traditional regression models. Method: From January 2017 to December 2021, 241 children with ASD, with an average age of 6.41 ± 1.96, diagnosed in the Developmental Behavior Department of the Children's Hospital Affiliated with the Medical College of Zhejiang University were included in the analysis. This study trained the traditional diagnostic models of Logistic regression (LR), Support Vector Machine (SVM), and two ensemble learning algorithms [Random Forest (RF) and XGBoost]. Socio-demographic and behavioral observation data were used to distinguish whether autistic children had combined ID. The hyperparameters adjustment uses grid search and 10-fold validation. The Boruta method is used to select variables. The model's performance was evaluated using discrimination, calibration, and decision curve analysis (DCA). Result: Among 241 autistic children, 98 (40.66%) were ASD comorbid ID. The four diagnostic models can better distinguish whether autistic children are complicated with ID, and the accuracy of SVM is the highest (0.836); SVM and XGBoost have better accuracy (0.800, 0.838); LR has the best sensitivity (0.939), followed by SVM (0.952). Regarding specificity, SVM, RF, and XGBoost performed significantly higher than LR (0.355). The AUC of ML (SVM, 0.835 [95% CI: 0.747-0.944]; RF, 0.829 [95% CI: 0.738-0.920]; XGBoost, 0.845 [95% CI: 0.734-0.937]) is not different from traditional LR (0.858 [95% CI: 0.770-0.944]). Only SVM observed a good calibration degree. Regarding DCA, LR, and SVM have higher benefits in a wider threshold range. Conclusion: Compared to the traditional regression model, ML model based on socio-demographic and behavioral observation data, especially SVM, has a better ability to distinguish whether autistic children are combined with ID.

[Characteristics of Xin'an medical masters in treatment of bi syndrome with acupuncture and moxibustion based on consolidating the root and cultivating the primary].

Based on the theory of consolidating the root and cultivating the primary, the paper collates and reviews the theoretical evidences and the characteristics of Xin'an medical masters in treatment of bi syndrome with acupuncture and moxibustion so as to provide the ideas for further research. Xin'an medical masters thoroughly acquainted with the theory of consolidating the root and cultivating the primary in treatment of bi syndrome with acupuncture and moxibustion, emphasizing the regulation of qi and blood, yin and yang, the nutrient qi and the defensive qi; and replenishing the middle jiao (spleen and stomach) and the lower jiao (kidney). The acupoint selection was distinctive, in which, the acupoints were selected and stimulated in terms of the etiology and the pathogenesis of diseases, as well as the properties of special points. The remarkable therapeutic effect on bi syndrome was ensured through specially selecting he-sea points, ashi points and "yin-yang opposite" points; effectively using the penetrating needling technique and moxibustion method and combining acupuncture with herbal medication.

Intelligence profiles and adaptive behaviors of high-functioning autism spectrum disorder and developmental speech and language disorders.

Objective: The present study was aimed at investigating the intelligence profiles and adaptive behaviors of children with high-functioning autism spectrum disorder (HFASD) and developmental speech and language disorders (DSLDs). We compared the similarities and differences of cognitive capabilities and adaptive functions and explored their correlations in the HFASD and DSLDs groups. Methods: 128 patients with HFASD, 111 patients with DSLDs and 114 typically developing (TD) children were enrolled into our study. Wechsler Intelligence Scale for Children-IV (WISC-IV) and Adaptive Behavior Assessment System-II (ABAS-II) were respectively applied to evaluate intelligence profiles and adaptive behaviors. Intelligence quotient (IQ) scores and adaptive functioning scores among the HFASD, DSLDs and TD groups were compared through one-way ANOVA. Pearson correlation coefficient was applied to examine the relationships between WISC indices and ABAS domains. Results: Outcomes showed significantly poorer intelligence profiles and adaptive behaviors in HFASD and DSLDs groups. Both children with HFASD and DSLDs demonstrated impairments in verbal comprehension and executive functions. Processing speed and working memory were the predominant defects of children with HFASD and DSLDs in the field of executive functions, respectively. Whereas perceptual reasoning was a relative strength for them. Children with DSLDs had balanced scores of all the domains in ABAS-II; nevertheless, HFASD individuals demonstrated striking impairments in Social domain. Correlation analysis showed IQs of children with HFASD were positively correlated with all the domains and General Adaptive Composite (GAC) of ABAS-II. Additionally, IQs were positively correlated with Conceptual domain and GAC for children with DSLDs. Compared with DSLDs group, intelligence displayed stronger correlations with adaptive behaviors in HFASD group. Conclusion: Our study expanded insights regarding intelligence profiles and adaptive behaviors of children with HFASD and DSLDs. Moreover, this study made breakthroughs in discovering positive correlations between IQs and adaptive functions in the two neurodevelopmental disorders.

Spatiotemporal expression of PSD-95 in Fmr1 knockout mice brain.

To investigate and compare the spatial and temporal expression of post-synaptic density-95 (PSD-95) in Fmr1 knockout mice (the animal model of fragile X syndrome, FXS) and wild-type mice brain, on postnatal day 7 (P7), P14, P21, P28 and P90, mice from each group were decapitated, and three principal brain regions (cerebral cortex, hippocampus and cerebellum) were obtained and stored for later experiments. PSD-95 mRNA in the three brain areas was analyzed with quantitative RT-PCR. PSD-95 protein was measured by immunohistochemical staining and Western blot. In the three principal brain areas of Fmr1 knockout mice and wild-type mice, the expression of PSD-95 mRNA and protein were detected at the lowest levels on P7, and then significantly increased on P14, reaching the peak levels in adolescents or adults. Moreover, it was found that PSD-95 mRNA and protein in the hippocampus were significantly decreased in Fmr1 knockout mice during the developmental period (P7, P14, P21 and P28) as well as at adulthood (P90) (P < 0.05, and P < 0.01, respectively). However, there was no significant difference of expression of PSD-95 in the cortex and cerebellum between Fmr1 knockout and wild mice. The expression of PSD-95 in the hippocampus might be regulated by fragile X mental retardation protein (FMRP) during mice early developmental and adult periods. It is suggested that impairment of PSD-95 is possibly involved in hippocampal-dependent learning defects, which are common in people with FXS.

Evaluation of Whole Blood Trace Element Levels in Chinese Children with Autism Spectrum Disorder.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder, which has increased markedly during the last decades. Essential trace elements play an important role in neurological function and their imbalances are common in children with ASD. The objective of the present study was to investigate whole blood levels of trace elements including zinc (Zn), copper (Cu), iron (Fe), and magnesium (Mg) in Chinese children with ASD. In total, 113 children diagnosed with ASD and 141 age-matched and gender-matched neurotypical children, divided into two gender and age groups of preschool age (2-5 years old) and school (6-10 years old) age, were examined. The quantitative analyses of whole blood trace element contents were performed by using flame atomic absorption spectroscopy. In the present study, the children with ASD generally had lower whole blood levels of Zn than the neurotypical controls. No significant differences in the whole blood Cu, Zn/Cu ratio, Fe, or Mg was detected between the ASD group and the control group. It is notable that whole blood Fe level in boys with ASD was significantly higher than in girls with ASD, and was nearly significant when compared with the control level of boys. After stratification for age, a significant 6% decrease in whole blood Zn levels was detected in preschool-aged children with ASD as compared to the control values. However, this significant ASD-related change was not detected in school-aged children. The whole blood Zn level and Zn/Cu ratio were significantly increased in school-aged children than in preschool-aged children in both ASD and control group. In addition, school-aged children with ASD had a significantly higher level of whole blood Fe than preschool-aged children with ASD. The results of the present study suggest an association between whole blood levels of Zn in Chinese children with ASD.

Absolute Quantification of Allergen Glb33 in Rice by Liquid Chromatography-Mass Spectrometry using Two Isotope-Labeled Standard Peptides.

Allergen Glb33 is an important allergen in rice that can cause allergic reactions such as asthma and atopic dermatitis. However, knowledge of the content in rice is sparse. In the present work, an absolute protein quantification method was established for allergen Glb33 in rice samples using liquid chromatography-tandem mass spectrometry. After extraction of allergen Glb33 from rice grains using salt solution, the isotope-labeled peptide internal standard was added to the extract, followed by enzymatic digestion with trypsin. The signature peptide and its isotope-labeled analogue from the tryptic hydrolysates of allergen Glb33 and the internal standard were detected by liquid chromatography-tandem mass spectrometry. The quantitative bias caused by tryptic efficiency and matrix effect was corrected by using two isotope-labeled standard peptides. The method exhibited good linearity in the range of 1-200 nM, with coefficients of determination of R2 > 0.998. A high sensitivity was observed, with a limit of quantification of 0.97 nM. Mean recoveries obtained from different rice matrices ranged from 82.7%-98.1% with precision <8.5% in intraday trials ( n = 6), while mean recoveries were from 75.1%-107.4% with precision <14.6% in interday trials ( n = 14). The developed method was successfully applied to the analysis of allergen Glb33 in 24 different rice cultivars.

Effects of pharmaceuticals on microbial communities and activity of soil enzymes in mesocosm-scale constructed wetlands.

Cyperus alternifolius based mesocosm-scale constructed wetland was employed to remove pharmaceuticals. We investigated the microbial community composition using phosphor lipid fatty acids (PFLAs) analysis and substrate enzyme activity during long-term exposure to pharmaceuticals in mesocosm-scale constructed wetlands. The results showed that there was no visible inhibition effect of pharmaceuticals on CW substrate enzymes activities in the experimental range (0-500 µg/L). Microbial communities, as revealed by PFLAs, were enhanced by the presence of plants, while the PFLAs content was highest when the pharmaceutical concentration was 10 µg/L or 30 µg/L at CWs. Except for anaerobic bacteria and Saturated fatty acids, the maximum PLFAs levels were reached when the pharmaceuticals were 10 µg/L or 30 µg/L, while Bacteria, G (-), fungal bacteria, Aerobic bacteria and Monounsaturated fatty acids were remarkably affected by high pharmaceuticals (100-500 µg/L). However, the main microbial florae were not changed among the treatments. In this study, the removal efficiencies of the studied pharmaceuticals in Planted (30) was greatest, which could be attributed to the higher microbial biomass. These results indicate that C. alternifolius can phytoremediate pharmaceutical-contaminated waters in CWs. Individual fatty acid cannot be used to represent specific species; therefore, more approaches to species identification such as rRNA-based methods must be included in future studies to better understand the metabolic mechanisms of microorganisms involved in the removal of studied pharmaceuticals and improve the performance of CWs.