Genotype and phenotype analysis in a case of cutis laxa type 3 / 中华内分泌代谢杂志

Objective:To report the clinical and genetic characteristics of autosomal dominant cutis laxa type 3 caused by ALDH18A1 mutation, and therefore to further understand this rare disease.Methods:High-precision full-exon sequencing was performed for the patient from the Department of Endocrinology and Genetic Metabolism, Children′s Hospital of Chongqing Medical University and genotype-phenotype correlation was summarized. Relevant literature was also reviewed.Results:A 9-month-old boy was admitted with complaint of " development retardation for 9 months, cough for 3 days" , accompanied by skin laxity, special features, skeletal malformation, tracheal bronchus, inguinal hernia, gastroesophageal reflux, and abnormal creases on palms. The heterozygous variation of ALDH18A1 c. 274C>G(p.Leu92Val) on chromosome 10 was revealed using high-precision full-exon sequencing. Together with imaging and metabolomics results, the diagnosis of cutis laxa type 3 was determined. The clinical presentations of this disease are variable, encompassing skin, bone, joint, and neuromuscular system.Conclusion:For suspected pediatric case, it is very important to evaluate the clinical manifestations and metabolic index at regular intervals, and to identify the molecular basis of the disease with gene sequencing early on.

DNA microarray screening analysis in children with profound hearing impairment in Hubei province / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate characteristics of molecular etiology of children with profound sensorineural hearing loss in Hubei province, and to provide reference for deafness treatment and genetic counseling.@*METHOD@#Three hundred and six children with profound sensorineural hearing loss in Hubei province were enrolled, their genomic DNA were extracted from peripheral blood and a deafness gene test chip was used to screen nine hot spot mutation in the GJB2, GJB3, SLC26A4, and mitochondria 12SrRNA gene. All patients with SLC26A4 gene mutation were given temporal bone CT scan.@*RESULT@#One hundred and thirty-two (43.14%) out of 306 children were found carrying at least one pathogenic gene mutation. The mutation rates of GJB2, SLC26A4 and mitochondria DNA 12SrRNA gene were 29.41% (90/306), 13.72% (42/306) and 0.65% (2/306), respectively. None out of 306 children was detected GJB3 gene mutation. Thirty-six patients carrying SLC26A4 gene mutation were detected enlarged vestibular aqueduct by CT scan.@*CONCLUSION@#Mutations of GJB2 and SLC26A4 gene are two major pathogenic gene for genetic hearing loss in children. 235delC mutation is the main mutation type, followed by IVS7-2A> G mutation type. The screening of SLC26A4 gene common mutations contribute to the diagnosis of enlarged vestibular aqueduct syndrome.

The study on 235delC mutation of GJB2 gene in patients with idiopathic sudden hearing loss / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To analyze the rate of 235delC mutation in GJB2 gene in patients with idiopathic sudden hearing loss, and to explore its possible correlation with pathogenesis of idiopathic sudden hearing loss.@*METHOD@#Two hundred and thirty-four patients with diagnosis of idiopathic sudden hearing loss in otolaryngology department were recruited as experimental group. Eighty people with normal hearing level were enrolled as control group. Their peripheral blood samples were obtained and genomic DNA was extracted. Using polymerase chain reaction, the coding region of GJB2 gene was amplified, and 235delC mutation is screened for in GJB2 gene by restriction endonuclease. At same time the clinical data of 234 patients was collected to analyze.@*RESULT@#In 234 cases of idiopathic sudden hearing loss, 5 cases were found to have heterozygous 235delC mutation, none of them harbored homozygous 235delC mutation, the 235delC mutation rate was 2.1% (5/234). No 235delC mutation was found in control group. The rate of 235delC mutation in two group showed no statistically significant difference (P > 0.05).@*CONCLUSION@#This research shows that the rate of 235delC mutation in GJB2 is low in patients with idiopathic sudden hearing loss, and suggest that 235delC mutation possible has no correlation with idiopathic sudden hearing loss.

Survey on quality of life in children and adolescents with type 1 diabetes / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To survey the quality of life in children and adolescents with type 1 diabetes.</p><p><b>METHODS</b>Ninety-eight children and adolescents with type 1 diabetes who participated in Diabetes Summer Camp held in Chongqing, Wuhan and Cheng during 2012 April and December were recruited in the study. The American juvenile diabetes patients quality of life scale Diabetes Quality of Life for Youths was used to assess the quality of life and SPSS19.0 was used for statistical analysis.</p><p><b>RESULTS</b>The scale had satisfactory reliability and validity with a Cronbach's Alpha score of 0.942 and a validity score of 0.679. All three dimension of scales: scales of impact, scales of worries and scales of satisfaction were significantly correlated with self-health assessment (P<0.01). The scores of impact and worries accounted for >50% of total scores as the same for the self health assessment scores. The score of disease course, diet and blood glucose control were positive correlated with each other. Age and HbA1c were positively correlated with the scale of impact, while gender has negative correlation with satisfaction scale (P<0.05). The diabetes diet had significant effects on the quality of life.</p><p><b>CONCLUSION</b>The quality of life in children and adolescents with type 1 diabetes is decreased, especially for those with longer disease course and female adolescents. The form of Diabetes Quality of Life for Youth used in the study has good reliability and validity, which can reflect the quality of life of Chinese diabetic children and adolescents.</p>

NADPH oxidase-dependent oxidative stress and mitochondrial damage in hippocampus of D-galactose-induced aging rats / 华中科技大学学报（医学）（英德文版）

Mitochondrial DNA (mtDNA) common deletion (CD) plays a significant role in aging and age-related diseases. In this study, we used D-galactose (D-gal) to generate an animal model of aging and the involvement and causative mechanisms of mitochondrial damage in such a model were investigated. Twenty 5-week-old male Sprague-Dawley rats were randomly divided into two groups: D-gal group (n=10) and control group (n=10). The quantity of the mtDNA CD in the hippocampus was determined using a TaqMan real-time PCR assay. Transmission electron microscopy was used to observe the mitochondrial ultrastructure in the hippocampus. Western blot was used to detect the protein levels of NADPH oxidase (NOX) and uncoupling protein 2 (UCP2). We found that the level of mtDNA CD was significantly higher in the hippocampus of D-gal-induced aging rats than in control rats. In comparison with the control group, the mitochondrial ultrastructure in the hippocampus of D-gal-treated rats was damaged, and the protein levels of NOX and UCP2 were significantly increased in the hippocampus of D-gal-induced aging rats. This study demonstrated that the levels of mtDNA CD and NOX protein expression were significantly increased in the hippocampus of D-gal-induced aging rats. These findings indicate that NOX-dependent reactive oxygen species generation may contribute to D-gal-induced mitochondrial damage.

NADPH oxidase-dependent oxidative stress and mitochondrial damage in hippocampus of D-galactose-induced aging rats / 华中科技大学学报（医学）（英德文版）

Mitochondrial DNA (mtDNA) common deletion (CD) plays a significant role in aging and age-related diseases. In this study, we used D-galactose (D-gal) to generate an animal model of aging and the involvement and causative mechanisms of mitochondrial damage in such a model were investigated. Twenty 5-week-old male Sprague-Dawley rats were randomly divided into two groups: D-gal group (n=10) and control group (n=10). The quantity of the mtDNA CD in the hippocampus was determined using a TaqMan real-time PCR assay. Transmission electron microscopy was used to observe the mitochondrial ultrastructure in the hippocampus. Western blot was used to detect the protein levels of NADPH oxidase (NOX) and uncoupling protein 2 (UCP2). We found that the level of mtDNA CD was significantly higher in the hippocampus of D-gal-induced aging rats than in control rats. In comparison with the control group, the mitochondrial ultrastructure in the hippocampus of D-gal-treated rats was damaged, and the protein levels of NOX and UCP2 were significantly increased in the hippocampus of D-gal-induced aging rats. This study demonstrated that the levels of mtDNA CD and NOX protein expression were significantly increased in the hippocampus of D-gal-induced aging rats. These findings indicate that NOX-dependent reactive oxygen species generation may contribute to D-gal-induced mitochondrial damage.

The recombinant adeno-associated viruses of the serotypes 2 mediated overexpression of MnSOD protects against oxidation-induced apoptosis in stria marginal cells of rat cochlea / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the influence of overexpression of manganese superoxide dismutase (MnSOD) of stria marginal cells (MCs) of the rat cochlea by the recombinant adeno-associated viruses of the serotypes 2 (AAV2) mediated gene-delivery for hydrogen peroxide-induced oxidative stress in vitro.@*METHOD@#Primary cultures of MCs were infected using rAAV2-MnSOD-EGFP at dosage of multiplicity of infection (MOI) 10(1)v x /cell and using rAAV2-EGFP as control. The expression of MnSOD in MCs was examined using western blot and the activity of MnSOD was determinated by colorimetric assays. Oxidative stress was induced in MCs by exposing them to H2O2 (400 micromol/L) for 2 hour and preculturing them in normal medium. After 24 h the amount of the lipid peroxidation production malondialdehyde (MDA) was detected. Apoptosis was assessed by flow cytometry by Propidium oidium staining. The expression of the cleaved Caspase-3 was assessed by Western blot.@*RESULT@#(1) EGFP expression in MCs could not be detected until 4 days after rAAV2- MnSOD-EGFP infection and reached fastigium after 10 days and lasted over a month. The MnSOD level in the rAAV2- MnSOD-EGFP group was higher than that in the control group. (2) After being exposed to H2O2, the amounts of MDA in rAAV2-MnSOD-EGFP group, control group and normal group were 0.464 +/- 0.049, 1.103 +/- 0.033 and 0.185 +/- 0.005 (nmol/mg prot), respectively. The expression of the cleaved-caspase-3 in rAAV2-MnSOD-EGFP group was lower than that in control group and the number of apoptotic cells decreased significantly.@*CONCLUSION@#The results demonstrate that the rAAV2-MnSOD-EGFP can effectively transfect cultured MCs, and the transgenic cells show a high expression of MnSOD which can protect the MCs against oxidative challenge. The role of overexpression MnSOD in MCs apoptosis induced by oxidative injury may be associated with suppressing the activation of caspase-3.

Protective roles of alpha-lipoic acid in rat model of mitochondrial DNA4834bp deletion in inner ear / 华中科技大学学报（医学）（英德文版）

The protective roles of alpha-lipoic acid in the rat model of mitochondrial DNA (mtDNA) 4834bp deletion in inner ear were investigated. Forty female Wistar rats at 4 weeks of age were divided into four groups: group A (D-galactose group, n=10), group B (D-galactose+alpha-lipoic acid group, n=10), group C (alpha-lipoic acid group, n=10), and group D (control group, n=10). Auditory brainstem response (ABR) was used to detect the hearing threshold. Colorimetry was used to analyze activity of superoxide dismutase (SOD) and concentration of malondialdehyde (MDA). The percentage of mtDNA4834bp deletion in inner ear was identified by real-time PCR. There was no significant difference in ABR threshold shift among all groups. The percentage of mtDNA4834bp deletion in group A was higher than that in other groups, but there was no significant difference in percentage of mtDNA4834bp deletion among groups B, C, and D. The activity of SOD in group A was lower than that in other groups. The concentration of MDA in group A was higher than that in other groups. It was concluded that there was no significant hearing loss when the percentage of mtDNA4834bp deletion was lower than 12.5%. alpha-Lipoic acid could prevent the reactive oxygen species (ROS)-induced mtDNA4834bp deletion in inner ear of rats.

The study of the high sensitivity to noise trauma in the inner ear mimetic aging model / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Use the inner ear mimetic aging model which has been established by our research institute, investigate its sensitivity to noise trauma and the possible role of mitochondrial DNA deletions.@*METHOD@#Thirty-two Wistar rats of 2 months old were randomly divided into four groups. In group A, D-galactose was subcutaneously injected at dose of 150 mg/kg weigh per day for 8 weeks, after that these rats were exposed to 110 dB SPL noise 4 hours each day, for 2 days. Group B were given normal saline (NS) injected at dose of 150 mg/kg weigh per day for 8 weeks,also given noise exposure as that of group A. Group C were give D-galactose without noise exposure. Group D were given normal saline (NS) without noise exposure. The thresholds of auditory brainstem response (ABR) were measured 2 weeks after stopping of noise exposure. And T-SOD and MDA of the inner membranous labyrinthine tissue were measured. Nest polymerase chain reaction (Nest PCR) were used to identify the mtDNA common deletion (CD), and PCR products were sequenced in the meantime.@*RESULT@#The elevation of the mean ABR thresholds in group A was higher than that in group B, and the difference had statistic significance (P < 0.01). The reduction of T-SOD in group A was obvious, while the level of MDA was greatly increased. The difference in the levels of T-SOD and MDA between group A and group B had statistic significance (P < 0.01). The detection rate of mtDNA 4834 deletion were as follows: group A 87.5% (7/8); group B 12.5% (1/8); group C 75.0% (1/8); group D 0(0/8).@*CONCLUSION@#The rat in the inner ear mimetic aging model are hypersensitive to noise exposure, and mtDNA4834 deletions in the inner ear may play an important role in it.

Expression change of mGluR5 in rat MVN after unilateral labyrinthectomy / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the expression of mGluR5 in the medial vestibular nucleus (MVN) following unilateral labyrinthectomy (UL).@*METHOD@#Thirty Wistar rats were randomly divided into two groups. Twenty four animals received unilateral labyrinthectomy while the others maintained labyrinthine well. After setting left labyrinthine, the change of mGluR5 was induced by immunohistochemistry, in situ hybridization.@*RESULT@#mGluR5 was increased in lesioned side MVN after unilateral labyrinthectomy. The 12 h post-UL was highest. Then it was decrease in 36 h post-UL, while 7 d post-UL was same as control group. The change of contralateral was same as that in ipsilateral.@*CONCLUSION@#UL can induce increase of mGluR5 in the MVN. The reduced resting discharge in the primary vestibular afferents or in the central vestibular neurons may be responsible for the change of mGluR5. However the significance of the change of mGluR1 in the vestibular compensation is still unknown.