Objective: To develop an HPLC method for the simultaneous determination of 3 ultraviolet absorbers (benzylidene camphor sulfonic acid, camphor benzalkonium methosulfate, 4-methylbenzylidene camphor) in cosmetics.Methods: After extracted by acetonitrile-methanol-ammonium acetate aqueous solution-tetrahydrofur(30∶35∶30∶5, v/v)(for lotion and milk) or acetonitrile-methanol-ammonium acetate aqueous solution(30∶20∶50, v/v)(for cream and powder), the UV absorbers were separated on a Thermo scientific C18 column (250 mm×4.6 mm,5 μm) at 35 ℃ with methanol, acetonitrile and 0.02 mol·L-1ammonium acetate aqueous solution (pH 5.15 adjusted by glacial acetic acid) as the mobile phase with gradient elution at the flow rate of 1.0 ml·min-1, and then analyzed by a DAD detector at the wavelength of 288 nm and 300 nm.Results: Benzylidene camphor sulfonic acid, camphor benzalkonium methosulfate and 4-methylbenzylidene camphor showed good linearity within the range of 12.26-98.06 μg·ml-1(r=0.999 9), 9.31-74.48 μg·ml-1(r=0.999 9) and 6.86-54.88 μg·ml-1(r=0.999 9), respectively.The limit of detection (LOD) and the limit of quantitation (LOQ) were 4.6, 6.4 and 1.6 ng and 17.4, 15.9 and 5.5 ng, respectively.The average recovery was 99.2%(RSD=1.58%), 99.8%(RSD=2.38%) and 99.2%(RSD=2.03%)(n=36), respectively.Conclusion: The method is simple, rapid, reproducible, accurate and reliable, and can be applied in the determination of ultraviolet absorbers in cosmetics.

Objective To determine the role and mechanism of peroxisome proliferator activated receptors (PPAR) α in a mouse model of D-galactosamine/lipopolysaccharide (D-GalN/LPS)-induced acute liver failure(ALF).Methods Firstly,C57BL/6 mice were randomly divided into control group(n =8),ALF 2h group(n =8),ALF 4h group (n =8),ALF 6h group (n =8).Secondly C57BL/6 mice were randomly divided into control group(n =8),ALF group(n =8),WY14643 group(n =8).To induce ALF,the mice were injected intraperitoneally with D-GalN (700 mg/kg) and LPS (10 μg/kg).WY14643 (6 mg/kg),the selective agonist of PPAR α,was administered via tail vein two hours prior to D-GalN/LPS exposure.Two,four,and six hours after D-GalN/LPS treatment in the first study,mice were anesthetized and blood was collected,6h after D-GalN/LPS treatment in the second study,blood was collected.The liver tissue was harvested for histology and mRNA extraction.Serum levels of ALT and AST were measured to evaluate the hepatic damage.Inflammatory cytokines (TNFα,IL-1β,IL-6) and chemokines (CXCL-1,CXCL-10) were detected by real-time quantitative PCR.Differential protein expression of p-NF-κBp65,p-JNK,p-ERK,p-p38 in inflammatory pathways was detected by Western blotting.Significance of inter-group differences was assessed by one-way ANOVA,and pairwise comparison was performed by the least significant difference test.Results The gene and protein expression of PPAR α were gradually reduced during the development of ALF.Compared with the model group,the liver architecture was better preserved almost with normal morphology in WY14643-treated mice.Serum ALT and AST levels in WY14643-treated group were significantly lower [ALT:(555 ±62)U/L vs (2 898 ±822) U/L,P ＜0.05; AST:(791 ±58) U/L vs (3 013 ±997)U/L,P ＜ 0.05].The expression of proinflammatory cytokines and chemokines was significantly suppressed during the activation of PPAR α.In the second study,the levels of gene expression of proinflammatory cytokines and chemokines were detected in control group,ALF group and WY14643 group respectively as followings:TNFα (0.161 ± 0.085,7.996 ± 1.068,3.346 ± 0.94,P ＜ 0.05),IL-1β(0.041 ±0.002,3.657 ±0.904,0.176±0.089,P＜0.01),IL-6 (0.018 ±0.008,1.762 ±0.589,0.163±0.0487,P ＜0.05),CXCL-1 (0.063 ±0.008,7.881 ±0.966,2.737 ±0.864,P ＜0.01),CXCL-10 (0.054 ±0.005,5.671 ±0.948,2.578 ±0.804,P ＜0.05).Conclusion Our findings first demonstrate that PPARα protects liver from injury in an ALF mouse model by suppressing inflammatory response,indicating PPARα as a potential new therapeutic target for ALF.

OBJECTIVE: To analyze the pathogenic variants of the KIF1A gene and its corresponding protein structure in an autism spectrum disorder (ASD) family trio carrying harmful missense variants in the KIF1A gene. METHODS: The peripheral blood DNA of the patient and his parents was extracted and sequenced using whole exome sequencing (WES) technology and verified by Sanger sequencing. Bioinformatics software SIFT, PolyPhen-2, Mutation Taster, and CADD software were used to analyze the harmfulness and conservation of variants. The Human Brain Transcriptome (HBT) database was used to analyze the expression of the KIF1A gene in the brain. PredictProtein and SWISS-MODEL were further used to predict the secondary structure and tertiary structure of KIF1A wild-type protein and variant protein. PyMOL V2.4 was utilized to investigate the change of hydrogen bond connection after protein variant. RESULTS: The WES sequencing revealed a missense variant c.664A>C (p.Asn222His) in the child's KIF1A gene, and this variant was a de novo variant. The harmfulness prediction results suggest that this variant is harmful. By analyzing expression level of KIF1A gene in the brain. It is found that KIF1A gene widely expressed in various brain regions during embryonic development. By analyzing the variant protein structure, the missense variant of KIF1A will cause many changes in the secondary structure of protein, such as alpha-helix, beta-strand, and protein binding domain. The connection of hydrogen bond and spatial structure will also change, thereby changing the original biological function. CONCLUSION The KIF1A gene may be a risk gene for ASD.
Autism Spectrum Disorder/genetics* ; Child ; Female ; Humans ; Kinesin/genetics* ; Mutation ; Mutation, Missense ; Pregnancy ; Protein Domains ; Whole Exome Sequencing

Objective: To explore susceptibility genes for autism spectrum disorders (ASD). Methods: Whole-exome sequencing was carried out for 60 family trios affected with sporadic ASD. Genetic variants discovered in over 10% of the patients were selected for genotype-phenotype correlation and pathway enrichment analysis using Phenolyzer software and metascape database. Combining gene-phenotypic scores, pathway-related genes associated with neural and neurite triggering were screened for the candidates. Results: A total of 170 common variants were found to be associated with the ASD phenotype. Among these, there was only one high-confidence gene [SHANK2 (0.8146)] and four medium-confidence genes [ERBB2 (0.1322), LAMC3 (0.1117), PPFIA4 (0.1059), DISC1 (0.1002)]. Twenty-pathways and four biological processes were found to be statistically significant by pathway enrichment analysis, which included neuron projection morphogenesis (GO: 0048812), regulation of neuroblast proliferation (GO: 1902692), modulation of excitatory postsynaptic potential (GO: 0098815), and dendrite morphogenesis (GO: 0048813). Twenty-one genes were found to be closely associated with neurological and neurite triggering, among which only SHANK2, ERBB2, and DISC1 had above-medium confidence correlation scores with the ASD phenotypes. Conclusion Abnormal neuron projection morphogenesis (GO: 0048812) may be closely related to the occurrence of ASD. SHANK2, ERBB2, and DISC1 are susceptibility genes for ASD.

OBJECTIVE: To carry out genetic testing for a patient with 45,X/46,XY mosaicism and autism spectrum disorder (ASD). METHODS: Peripheral blood samples of the patient and his parents were collected for the extraction of genomic DNA. Trio-based whole exome sequencing and Sanger sequencing were carried out thereafter. RESULTS: The proband and his father were found to harbor a heterozygous c.4781G>A (p.Arg1594Gln) variant of the CACNA1I gene. In addition, the proband was also found to harbor a de novo c.268C>T (p.Arg90Trp) missense variant of the MTRR gene. Based on guidelines of the American College of Medical Genetics and Genomics (ACMG), the c.4781G>A (p.Arg1594Gln) variant of the CACNA1I gene was predicted to be pathogenic (PVS1, PM1, PM2, PP3), while the c.268C>T (p.Arg90Trp) variant of the MTRR gene was predicted to be of uncertain significance. CONCLUSION Variants of the CACNA1I and MTRR genes, together with the chromosomal mosaicism, may have predisposed to the susceptibility to the ASD in this patient.
Autism Spectrum Disorder/genetics* ; Genomics ; Heterozygote ; Humans ; Mosaicism ; Whole Exome Sequencing