ABSTRACT
Objective:To study the different effects of pre-pregnancy obesity (PO), excessive gestational weight gain (EGWG), pre-pregnancy obesity combined with excessive gestational weight gain (PO+EGWG) of maternal rats on glucose and lipid metabolism in neonatal offspring, and to explore the possible mechanisms.Methods:Animal models of PO, EGWG and PO+EGWG were established by feeding SD rats with high-fat diets at different periods. Thirty-six SD rats were randomly divided into four groups, with nine rats in each group. The control group had a normal diet before and during pregnancy. The PO group had a high-fat diet before pregnancy and a normal diet during pregnancy. The EGWG group had a normal diet before pregnancy and a high-fat diet during pregnancy. And the PO+EGWG group had a high-fat diet before and during pregnancy. The body weight of maternal rats before and during pregnancy and the birth weight of neonatal rats were recorded. Nine male neonatal rats in each group were selected, fasting blood glucose levels were detected by glucometer, fasting insulin levels were detected by enzyme-linked immunosorbent assay kit, hepatic triglyceride and cholesterol levels were detected by glycerol phosphate oxidase-peroxidase method, hepatic lipid deposition were observed by hematoxylin-eosin staining and oil red O staining. The mRNA levels of hepatic key genes in glucose metabolism pathway IR, IRS, AKT and lipid metabolism FASN, SREBP1c, PPARα were detected by reverse transcription-polymerase chain reaction analyses.Results:The pre-pregnancy weight of maternal rats in high-fat diet group before pregnancy (PO group and PO+EGWG group) was significantly higher than those in normal diet group (control group and EGWG group). The percentage of weight gain of maternal rats in high-fat diet group during pregnancy (EGWG group and PO+EGWG group) was significantly higher than those in normal diet group (control group and PO group) ( P<0.05). The birth weight of neonatal rats in PO group, EGWG group and PO+EGWG group were significantly higher than that in control group ( P<0.05), and the birth weight of neonatal rats in PO+EGWG group was the largest. The fasting glucose, insulin level and insulin resistance index of newborn rats in PO, EGWG and PO+EGWG groups were higher than those in the control group, and the mRNA levels of IR, IRS and AKT were lower than those in the control group, but the differences were not statistically significant ( P>0.05). The hepatic triglyceride and cholesterol contents and mRNA levels of FASN and SREBP1c were higher in the EGWG and PO+EGWG groups than those in the control group, and the mRNA level of PPARα was higher in the PO+EGWG group than in the control and PO groups, with statistically significant differences ( P<0.05). Conclusions:Animal models of PO, EGWG and PO+EGWG were successfully constructed by feeding SD rats with high-fat diets before pregnancy, during pregnancy, before and during pregnancy. PO+EGWG had the most significant effects on the birth weight and glucose and lipid metabolism in neonatal offspring. Compared with EGWG, PO had a relatively significant effect on glucose metabolism in neonatal offspring. And compared with PO, EGWG had a relatively significant effect on lipid metabolism in neonatal offspring. The effects of maternal obesity on glucose and lipid metabolism in neonatal offspring were considered to be related to the expression changes of genes in glucose and lipid metabolism.
ABSTRACT
Neonatal inherited metabolic diseases (IMD) screening has been widely conducted worldwide. Tandem mass spectrum (MS/MS) is the main procedure of IMD screening. As a new technique, gene sequencing has been put into practice for IMD screening. Nowadays, the morbidity and disease spectrum of IMD in China is still unclear. A summary of general and single morbidity, and disease spectrum of China's IMD from publications of MS/MS screening could provide evidence for establishing neonatal IMD's genetic test and formulation of laws and regulations.
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Childhood inherited leukodystrophies are common and diagnostic challenge disorders in inherited metabolic diseases.The rate of definite diagnosis of these diseases is lower,the early diagnosis is difficult,and the clinicians lack awareness for it.With the development of head imaging technology and the application of high-throughput gene sequencing technology,the list of inherited leukodystrophies has been increased to more than one hundred typies.Now,the classification,clinical features,specific biochemical tests and imaging characteristics of inherited leukodystrophies are reviewed,and the application of high-throughput sequencing in the diagnosis of these diseases in order to improve understanding of childhood inherited leukodystrophies are introduced.
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Objective To explore the excitotoxic role of NMDA receptors in striatal neurons in glutaric aciduria type I (GA1). Methods A GA1 cell model was established by lentivirus-mediated shRNA to GCDH and excessive intake of lysine. The expression levels of NMDA receptors were determined by Western blotting. The striatal neurons were preprocessed by MK801(a NMDA receptor antagonist), then infected with lentivirus and cultured in high concentration lysine. Cell viability was measured using MTT. Apoptosis was assessed using Hoechst33342 staining. Results Compared with the control group, the expression of NR2B protein in the experimental group was increased, and there was statistical difference (P<0.001). The differentces in the cell viability and normal nuclear proportion among experimental group, control group, and MK-801 pretreatment group were statistically significant (P<0.01). The cell viability and normal nucleus proportion in experimental group were significantly lower than those in control group while they were significantly higher in MK-801 pretreated group than those in the experiment group but still significantly lower than those in control group (P all <0.05). Conclusion The accumulation of metabolites in GA 1 played a toxic role in striatal neurons through NMDR receptors.
ABSTRACT
Objective To explore the excitotoxic role of NMDA receptors in striatal neurons in glutaric aciduria type I (GA1). Methods A GA1 cell model was established by lentivirus-mediated shRNA to GCDH and excessive intake of lysine. The expression levels of NMDA receptors were determined by Western blotting. The striatal neurons were preprocessed by MK801(a NMDA receptor antagonist), then infected with lentivirus and cultured in high concentration lysine. Cell viability was measured using MTT. Apoptosis was assessed using Hoechst33342 staining. Results Compared with the control group, the expression of NR2B protein in the experimental group was increased, and there was statistical difference (P<0.001). The differentces in the cell viability and normal nuclear proportion among experimental group, control group, and MK-801 pretreatment group were statistically significant (P<0.01). The cell viability and normal nucleus proportion in experimental group were significantly lower than those in control group while they were significantly higher in MK-801 pretreated group than those in the experiment group but still significantly lower than those in control group (P all <0.05). Conclusion The accumulation of metabolites in GA 1 played a toxic role in striatal neurons through NMDR receptors.
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Objective To study the NR5A1 gene mutation in patients with idiopathic hypogonadotropic hypogonadism(IHH), and to find the new mutation point.Methods Sixty-one IHH patients and 100 normal control subjects were collected and genomic DNA was extracted from blood samples.These patients were with normal karyotype and no abnormality was discovered in magnetic resonance imaging (MRI) scan of the pituitary.Other endocrine diseases were also excluded.The 2-7 exons and splice-sites of NR5A1 gene were amplified with polymerase chain reaction.DNA of the coding sequence and splice-sites of NR5A 1 were sequenced by double deoxidizing terminal end sequencing method in 61 IHH and 100 normal control subjects.The results of sequencing were compared with their corresponding sequence data.61 IHH kindreds were investigated and the clinical data of these patients were collected.Finally, the phenotype and genotype positive cases were analyzed.Results Six patients carried NR5A1 gene mutational sites in 61 cases of IHH.Analysis of sequencing results from 100 age and ethnicity matched control subjects did not show any of these novel changes.Conclusions One mutation in NR5A1 gene may affect protein structure and function, which should be considered in male IHH patients with normal karyotype and without insufficiency of adrenal function.
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Objective To investigate the expressions of insulin receptor substrate-1 (IRS-1) and insulin receptor sub-strate-2 (IRS-2) in adipocytes during catch-up growth in neonatal rats with intrauterine growth restriction (IUGR) and their correlations with the insulin resistance. Methods Sprague-Dawley rats (clean grade) were randomly divided into control group and food-restricted group after fertilization. Food-restricted group were received about 30%of food amount consumed in control group every day through the whole pregnant period to establish IUGR animal model, and were fed increased amount of breast-milk from postnatal day 1 to 21 to establish the period of catch-up growth in IUGR animal model (IUGR-CG). Fasting serum glu-cose, insulin and triglyceride were measured in blood from heart ventricles of 4-week old SD rats and insulin resistance index was calculated. Pre-adipocytes and mature adipocytes were obtained from SD rats at different age (1-week, 3-week, 5-week and 7-week old) and the former were induced to differentiate toward mature adipocytes. The levels of IRS-1, IRS-2 in the two kinds of mature adipocytes were detected by Real-Time PCR and Western blot. Results The expression levels of IRS-1, IRS-2 mRNA in mature adipocytes of IUGR-CG rats were signiifcantly lower than those of IUGR rats at 5-weeks and 7-weeks old (P<0.05) while the ex-pression levels of IRS-1, IRS-2 mRNA in differentiated adipocytes of IUGR-CG rats were signiifcantly lower than those of IUGR rats at 5-weeks old (P<0.05). The expression levels of IRS-1, IRS-2 protein in two kinds of adipocytes (mature and differentiated adipocytes) of IUGR-CG rats were signiifcantly lower than those of IUGR rats from postpartum week 1 through 7 (P<0.05). Conclusions IRS-1 and IRS-2 expression levels are downregulated in adipocytes during catch-up growth of IUGR rats, which may be closely related with insulin resistance.