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To retrospectively explore the characteristics of plasma amino acids (PAAs) in children with autism spectrum disorder and their clinical association via case-control study. A total of 110 autistic and 55 healthy children were recruited from 2014 to 2018. The clinical phenotypes included severity of autism, cognition, adaptability, and regression. Compared with the control group, autistic children had significantly elevated glutamate, γ-Amino-n-butyric acid, glutamine, sarcosine, δ-aminolevulinic acid, glycine and citrulline. In contrast, their plasma level of ethanolamine, phenylalanine, tryptophan, homocysteine, pyroglutamic acid, hydroxyproline, ornithine, histidine, lysine, and glutathione were significantly lower. Elevated neuroactive amino acids (glutamate) and decreased essential amino acids were mostly distinct characteristics of PAAs of autistic children. Increased level of tryptophan might be associated with severity of autism.

The SLC25A32 dysfunction is associated with neural tube defects (NTDs) and exercise intolerance, but very little is known about disease specific mechanisms due to a paucity of animal models. Here, we generated homozygous (Slc25a32Y174C/Y174C and Slc25a32K235R/K235R) and compound heterozygous (Slc25a32Y174C/K235R) knock-in mice by mimicking the missense mutations identified from our patient. A homozygous knock-out (Slc25a32-/-) mouse was also generated. The Slc25a32K235R/K235R and Slc25a32Y174C/K235R mice presented with mild motor impairment and recapitulated the biochemical disturbances of the patient. While Slc25a32-/- mice die in utero with NTDs. All Slc25a32 mutations did not hinder the mitochondrial uptake of folates but specifically blocked the uptake of flavin adenine dinucleotide (FAD). A positive correlation between SLC25A32 dysfunction and flavoenzyme deficiency was observed. Besides the flavoenzymes involved in fatty acid β-oxidation and amino acid metabolism impaired, Slc25a32-/- embryos still had a subunit of glycine cleavage system–dihydrolipoamide dehydrogenase damaged, resulting in glycine accumulation and glycine derived-formate reduction, which further disturbed folate-mediated one-carbon metabolism, leading to 5-methyltetrahydrofolate shortage and other folate intermediates accumulated. Maternal formate supplementation increased the 5-methyltetrahydrofolate levels and ameliorated the NTDs in Slc25a32-/- embryos. The Slc25a32K235R/K235R mice had no glycine oxidation defect but had another formate donor-choline metabolism interrupted and mitochondrial folates deficient. Formate supplementation increased mitochondrial folates amounts of mice, but this effect was not restricted to the Slc25a32 mutant mice. In summary, we established novel animal models, which enabled us to better understand the function of SLC25A32 and to elucidate the role of SLC25A32 dysfunction in human disease development and progression.

Stress plays an important role in reproductive health and likely is one of the psychological factors affecting ART success. This study was designed to examine the relationship between the stress level as inferred from the amount of the enzyme alpha-amylase secreted in saliva (SAA) and pregnancy outcome in infertile couples undergoing in vitro fertilization and embryo transplantation (IVF-ET). A prospective cohort study was conducted in the Reproductive Medicine Centre of Zhengzhou University Hospital in Henan, China. Four hundred fifty-seven infertile couples undergoing in vitro fertilization and embryo transplantation (IVF-ET) for the first time participated in the study. Couples collected saliva samples the morning before the start of their first treatment cycle for the measurement of SAA. We found that the level of SAA (and hence, the amount of stress) in female partners, male partners, and couples analyzed together significantly affected IVF-ET outcome. Cutoff levels of SAA that predicted pregnancy failure were 136 μmol/L, 149 μmol/L, and 288 μmol/L in female partners, male partners, and couples, respectively. Female partners, male partners, and couples with high SAA levels had increased risk of pregnancy failure compared to those with low SAA levels. The SAA level directly correlated with the follicle-stimulating hormone level and was inversely proportional to the anti-Müllerian hormone level and endometrial thickness. Some semen parameters of male partners, such as density, survival rate, sperm rapid progressive motility (A%), and progressive motility [(A + B)%], were significantly lower in the high-SAA than in the low-SAA group. Furthermore, couples in the high SAA group had fewer transferable and high-quality embryos. We concluded that a high SAA level, known to be an objective indicator of high stress, increases the risk of pregnancy failure in infertile couples undergoing IVF-ET. Lay summary To explore the relationship between stress, as measured by the levels of the stress biomarker salivary alpha-amylase (SAA), and pregnancy outcome in infertile couples undergoing in vitro fertilization, a prospective cohort study was conducted in the Reproductive Medicine Centre of Zhengzhou University Hospital in Henan, China. Four hundred fifty-seven infertile couples undergoing IVF-ET collected saliva samples the morning before the start of their first treatment cycle for the measurement of SAA. Results of this study demonstrated that a high SAA level, known to be an objective indicator of high stress, increases the risk of pregnancy failure in infertile couples undergoing IVF-ET.

To study the molecular genetic mechanism and genetic diagnosis of pyruvate dehydrogenase complex deficiency (PHD), and to provide a basis for genetic counseling and prenatal genetic diagnosis of PHD.Polymerase chain reaction (PCR) was performed to amplify the 11 exons and exon junction of the PDHA1 gene from a child who was diagnosed with PHD based on clinical characteristics and laboratory examination results. The PCR products were sequenced to determine the mutation. An analysis of amino acid conservation and prediction of protein secondary and tertiary structure were performed using bioinformatic approaches to identify the pathogenicity of the novel mutation.One novel duplication mutation, c.1111_1158dup48bp, was found in the exon 11 of the PDHA1 gene of the patient. No c.1111_1158dup48bp mutation was detected in the sequencing results from 50 normal controls. The results of protein secondary and tertiary structure prediction showed that the novel mutation c.1111 _1158dup48bp led to the duplication of 16 amino acids residues, serine371 to phenylalanine386, which induced a substantial change in protein secondary and tertiary structure. The conformational change was not detected in the normal controls.The novel duplication mutation c.1111_1158dup48bp in the PDHA1 gene is not due to gene polymorphisms but a possible novel pathogenic mutation for PHD.

To study the relationship of karyotypes and gonadal development in children with Turner syndrome.Fifty-one children with Turner syndrome were studied. Pelvic ultrasonography was performed on the children. Twenty healthy age-matched girls were used as control group.Eleven kinds of karyotypes were detected in 51 children with Turner syndrome. Children were classified into two groups based on karyotypes: Group 1 (45,XO; n= 24) and Group 2 (karyotypes other than 45, XO; n=27). 45,XO karyotype was the most common (47.1%), followed by 46,X,i (Xq)/45, XO (21.6%), 46,XX/45, XO (7.8%) and 47,XXX/45, XO (5.9%). Pelvic ultrasonography showed a primordial uterus in 41 cases, infantile uterus in 7 cases, congenital absence of uterus and ovary in 3 cases, simple anovarism in 42 cases and ovarian dysgenesis in 6 cases. Uterine size in both Groups 1 and 2 were significantly smaller than those of the control group (P0.05). Group 1 showed smaller size of uterus than Group 2 (P0.05). Ovaries were not detected in all Group 1 patients.The dysplasia and atrophy of ovaries and uterus exist in children with Turner syndrome. The patients with 45,XO karyotype had poorer gonadal development than those with other karyotypes.

Female precocious puberty is caused by premature activation of the hypothalamic-pituitary-gonadal axis, exposure to exogenous sex steroid hormones, and the presence of endogenous sex steroids caused by various factors. Estrogen is the final key factor to start onset of puberty. However,in some cases of precocious puberty in girls estrogen elevation could not be detected. The raised sensitivity of estrogen receptor, which may caused by ESR1 mutation or polymorphism, has been frequently mentioned for interpreting the etiology of sporadic low estrogen type cases. But no case evidence has been found in clinical practice. For the purpose of screening possible mutations in estrogen receptor gene, leukocyte genomic DNA were collected from 16 girls with precocious puberty of sporadic low estrogen,and exons of ESR1 were amplified and analysized using PCR-SSCP/silver staining method. A single strand conformation change in exon 8 was found in one of the patients (No. 14). The suspected fragment were cloned to a T vector and sequenced for analysis. Sequencing of these clones revealed that this conformation change is caused by a C to T transition. This mutation results in the replacement of arginine by cystine at position 548 of ESR1 protein. The mutation created an extra Btsl digest site and made it can be readily identified by PCR-PFLP method. Further detection using this method, and sequencing of cloned exon8 colonies from patients proved that the patient No. 14 is Arg548/Cys548 heterozagous in genotype. This mutation increased hydrophobility of the area dramatically. The position and the conservative of this residue in vertebrates suggested Arg548 may play an important role in ESR1 function. For study the role of this mutation in the onset of precocious puberty, a firefly luciferase reporter plasmid pGL3-promoter-ERE was constructed,and a pCR3. 1-hermut pisimid expressing Cys548 ER was constructed based on wild type pCR3. 1her. Co-transfection of reporter and pCR3. 1 -hermut in CMF-7 cell strain proved that Cys548 mutant can significantly increase the transcription activity over the Arg548 wild type.