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Objective@#To investigate the physical and intellectual development and mutation characteristics of the phenylalanine hydroxylase (PAH) gene among 53 newborns with phenylketonuria (PKU), so as to provide insights into the management and genetic counseling of PKU@*Methods@#The medical records of 54 children with definitive diagnosis of PKU and standardized therapy until 2 years at the Center for Neonatal Disease Screening of Shanxi Children' s Hospital from 2018 to 2021 were collected. Newborns' body weight and height developments were evaluated using the World Health Organization growth chart (2006 version), and the intellectual development was assessed using the national criteria of Development Behavior Assessment Scale among Children at Ages of 0 to 6 Years (WS/T 580-2017). The gene mutations were detected among neonates and their children, and the physical, intellectual developments and genetic characteristics of neonates with PKU were descriptively analyzed.@*Results@#The 53 PKU cases included 29 male children and 24 female children, 36 cases with classic PKU and 17 cases with mild PKU, and 30 cases from rural areas and 23 cases from urban areas. The study subjects had a median age of 30 (10) d at initial therapy, and a mean blood phenylalanine concentration of (1 507±685) μmol/L at definitive diagnosis. There were 52 cases with normal height developments (98.11%), and all cases had normal weight and intellectual developments. The mean developmental functional quotient (DFQ) was significantly greater among urban children with PKU than among rural children [(94.92±8.57) vs. (87.65±6.57); t=-3.498, P=0.001], and the mean DFQ was significantly higher among children with mild PKU than among those with classic PKU [(95.55±8.76) vs. (88.57±7.11); t=-3.095, P=0.003]. There were 37 mutations detected in the PAH gene, which were mainly distributed in exons 3, 6, 7, 11, 12 and intron 4. Three high-frequency mutation sites were detected, including c.728G>A, c.611A>G and c.1197A>T, including three novel mutations (c.674C>G, c.1316-2A>C and c.1069T>C).@*Conclusions@#Following standardized treatment, the children with PKU have comparable physical and intellectual developments as compared to normal children. c.728G>A, c.611A>G and c.1197A>T were predominant mutations in the PAH gene among these 53 children with PKU, and three novel mutations were identified, including c.674C>G, c.1316-2A>C and c.1069T>C.
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Objective:To investigate the distribution of clustered regularly interspaced short palindromic repeats (CRISPR) in group B Streptococcus (GBS) in the genital tract of women during the third trimester and in infants with invasive infection and its relationship with multilocus sequence typing (MLST) and drug-resistance genes. Methods:This study retrospectively collected 84 GBS strains isolated from pregnant women with GBS colonization and infants with invasive GBS infection who were admitted to Children's Hospital Affiliated to Shanxi Medical University from January 2017 to January 2022. CRISPR, MLST, and drug-resistance phenotype and genes were detected and analyzed using χ 2 test or Fisher exact probability method. MEGA11 was used to construct a dendrogram. Results:There were ten sequence typing in the 84 GBS strains and ST10 was the dominant one (46.4%). GBS was sensitive to penicillin, and its resistance rates to erythromycin (75.0%) and clindamycin (73.8%) were high. Among the 17 invasive GBS strains, ST10 had 100% resistance to erythromycin, clindamycin, and levofloxacin. CRISPR1 gene was amplified in 62 strains (73.8%). CRISPR1-positive strains had a significantly higher proportion of ST10 [56.5%(35/62) vs 18.2%(4/22), χ 2=9.56, P=0.002] and ermB, gyrA, parC [54.8%(34/62) vs 22.7%(5/22), 67.7%(42/62) vs 36.4%(8/22), 71.0%(44/62) vs 36.4%(8/22); χ 2=6.73, 6.64, and 8.25, all P<0.05], and a lower proportion of ermA [6.5%(4/62) vs 31.8%(7/22), χ 2=7.09, P=0.008] than CRISPR1-negative strains. Conclusions:ST10 is the main GBS genotype among the colonized microbiota the genital tract of pregnant women and in infants with invasive GBS infection, which is also a dominant type in CRISPR1-positive strains. GBS is sensitive to penicillin and CRISPR1 gene is linked to the spread of some drug-resistance genes.
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Objective:To investigate the effect and mechanism of silent information regulator 6 (SIRT6) and gluconeogenesis-dependent rate-limiting enzymes in hepatocytes in oxidative stress injury rats and chronic-on-acute (sub-acute) liver failure (ACLF) patients.Methods:From August 2016 to May 2018, 10 patients with ACLF from Beijing Youan Hospital Affiliated to Capital Medical University were included in the ACLF group, and 10 normal donors were included in the normal control group. Level of fasting blood glucose, total bilirubin, albumin, and alanine aminotransferase (ALT) were studied. Sprague Dawley rat hepatocytes were isolated and divided into control group (without any intervention), model group (H 2O 2 intervention for 6 h), mammalian rapamycin target protein (mTOR) activation group (mTOR activation was added to the model group), mTOR inhibition group (mTOR inhibitor was added on the basis of the model group). Protein electrophoresis and polymerase chain reaction was used to detect the relative expression of glucose-6-phosphatase (G6P), phosphoenolpyruvate (PEPCK), SIRT6, and mTOR. Results:The ALT and total bilirubin level in ACLF group were significantly higher than those in the normal control group, and the differences were statistically significant (all P<0.05). In ACLF group, level of SIRT6 (0.15±0.07) μg/L and fasting blood glucose (3.19±0.59) mmol/L were significantly lower than those in the normal control group (0.46±0.15) μg/L and (7.07±2.07) mmol/L, the difference was statistically significant (all P<0.05). The relative expression of PEPCK and G6P protein in liver tissue of ACLF group was significantly lower than that of normal control group. The relative expression of SIRT6, PEPCK, and G6P in the model group were lower than those in the control group, and the differences were statistically significant (all P<0.05). When mTOR is activated, the relative expression of PEPCK, G6P, and SIRT6 was higher than those in the model group, and after mTOR inhibition, the relative expression of PEPCK, G6P, and SIRT6 was lower than in the model group. Conclusion:ACLF, SIRT6 may inhibit gluconeogenesis, and increased the occurrence of hypoglycemia through activating mTOR signaling pathway. Blocking of SIRT6 levels may slow down the progress of ACLF.