A novel mutation in the ABCD1 gene causes adrenomyeloneuropathy by impairing autophagy level / 中华神经科杂志

Objective:To report a Chinese family with a novel ABCD1 gene mutation at c.332T>G (p.V111G) site and discuss its clinical characteristics and molecular mechanism.Methods:The clinical data, laboratory examination, and imaging examination results were analyzed to make the clinical diagnosis of a middle-aged onset patient from the First Affiliated Hospital of Zhengzhou University in May 2017. High-throughput sequencing was used to discover a novel ABCD1 gene mutation. Sanger sequencing was used to find out whether other family members contain the same ABCD1 gene mutation. The pathogenicity of this mutation was explored by protein structure prediction and pathogenicity analysis. Adrenoleukodystrophy protein-green fluorescent protein (ALDP-GFP) and ALDP-GFP (V111G) plasmids were constructed and human embryonic kidney 293 cells were transfected, then immunofluorescence and Western blotting were used to explore the molecular mechanism of this mutation (completed in Henan Provincial People′s Hospital).Results:The proband (a 39-year-old male) was diagnosed as adrenomyeloneuropathy, a subset of X-linked adrenoleukodystrophy, with a novel heterozygous missense mutation in the ABCD1 gene at c.332T>G (p.V111G) site, and his mother and two daughters were all carriers. Protein structure prediction and pathogenicity results suggested that this mutation is pathogenic. Overexpression of ALDP-GFP (V111G) in the human embryonic kidney 293 cells resulted in a significant decrease in the expression levels of ALDP and the abnormal localization from the peroxisomal membrane to the cytoplasm, accompanied by significant down-regulation of LC3-Ⅱ/LC3-Ⅰ and beclin-1.Conclusion:c.332T>G (p.V111G) is a novel pathogenic mutation in the ABCD1 gene, which causes adrenomyeloneuropathy by impairing autophagy.

Genetic diagnosis of a child with aortic stenosis and thumb aplasia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the molecular mechanism and prognosis of a child with aortic stenosis and thumb aplasia.</p><p><b>METHODS</b>The karotypes of the child and his parents were analyzed with routine G-banding. Their genomic DNA was also analyzed with array comparative genomic hybridization(aCGH) for chromosomal duplications/deletions.</p><p><b>RESULTS</b>No karyotypic abnormality was detected at cytogenetic level for the child and his parents. aCGH identified a de novo 5.86 Mb deletion at 2q22.3-q23.3 in the child.</p><p><b>CONCLUSION</b>The child was diagnosed with 2q23.1 microdeletion syndrome. MBD5 may be the key gene for the 2q23.1 microdeletion syndrome.</p>

Genetic and prenatal diagnosis for a haemophilia A family with two novel mutations of F8 gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To conduct genetic diagnosis for a family affected with hamophilia A.</p><p><b>METHODS</b>Potential mutations of the F8 gene were analyzed with PCR and Sanger sequencing. Carriers of the mutation were identified through linkage analysis using short tandem repeat (STR) markers. Suspected mutations were verified among 100 healthy controls to rule out genetic polymorphism. Prenatal diagnosis was provided based on the above results.</p><p><b>RESULTS</b>Sequencing analysis has identified two mutations, c.1 A>T and c.4 C>T, which have replaced the start codon (ATG) with leucine (TTG) and glutamine (GAA) with the stop codon (TAA), respectively. The same mutations were not found among the 100 healthy controls. The patient's mother and sister were heterozygous for the same mutations. Upon prenatal diagnosis, the fetus was determined as a male and did not harbor the above mutations. Linkage analysis also confirmed that the fetus has inherited the non-risk X chromosome from his maternal grandfather.</p><p><b>CONCLUSION</b>Detection of pathogenic mutations can enable prenatal diagnosis for the disease.</p>

Genetic diagnosis for a family without exonic deletions and duplications of dystrophin gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To conduct genetic diagnosis for a family in which no exonic deletions and duplications of the dystrophin gene were detected.</p><p><b>METHODS</b>Potential exonic deletions and duplications of the dystrophin gene were initially analyzed with using multiplex ligation-dependent probe amplification (MLPA). Subsequently, all of the 79 exons of the dystrophin gene of the proband and a pregnant woman from the family were analyzed with PCR amplification and DNA sequencing. Following identification of the causative mutation, prenatal diagnosis was provided.</p><p><b>RESULTS</b>MLPA analysis had detected no exonic deletions and duplications of the dystrophin gene. Sequence analysis has identified a C>T mutation on the 22nd nucleotide position of the 70th exon of the dystrophin gene (c.10108 C>T), which has replaced the codon CGA to a stop codon (TGA). The patient's mother and sister were both heterozygous for the same mutation. Upon prenatal diagnosis, the fetus was found to be positive for the Y chromosome sex-determining gene (SRY) and has carried above mutation. The result of short tandem repeat linkage analysis also confirmed that the fetus has inherited the mutant X chromosome.</p><p><b>CONCLUSION</b>The causative mutation of the dystrophin gene has been discovered in an affected family, which has enabled prenatal diagnosis of the disease.</p>

Predictive value of HLA-DRB1 gene for the treatment of unexplained recurrent spontaneous abortion with paternal lymphocyte alloimmunization therapy in Henan Hans / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the value of HLA-DRB1 gene in predicting the outcome of unexplained recurrent spontaneous abortion (URSA) treated with paternal lymphocyte alloimmunization therapy (PLAT) in Henan Hans.</p><p><b>METHODS</b>Three hundred URSA patients were recruited. Following PLAT treatment, they were divided into two groups according to the outcome of pregnancy. Polymerase chain reaction sequence specific primer (PCR-SSP) were conducted to analyze the HLA-DRB1 gene.</p><p><b>RESULTS</b>For those who have received PLAT treatment, the frequency of HLA-DRB1*11 was significantly lower in successfully treated cases than those with abortion (0.052 vs. 0.110, P < 0.05, OR=0448), whilst the frequency of HLA-DRB1*15 was significantly greater in the former (0.207 vs. 0.100, P < 0.05, OR=2.352).</p><p><b>CONCLUSION</b>For patients who have received PLAT treatment, those with HLA-DRB1*15 are more likely to conceive that those with HLA-DRB1*11.</p>

Association of polymorphisms of HLA-DRB1 gene with unexplained recurrent spontaneous abortion in ethnic Hans from Henan / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of polymorphisms of human leukocyte antigen DRB1 gene (HLA-DRB1) with susceptibility to unexplained recurrent spontaneous abortion (URSA).</p><p><b>METHODS</b>The HLA-DRB1 gene was typed with polymerase chain reaction-specific sequence primers (PCR-SSP) method in 200 couples with URSA and 200 couples with a normal pregnancy history.</p><p><b>RESULTS</b>The frequencies of DRB1*09 and DRB1*13 alleles were significantly greater in the URSA group compared with the control group (14.50% vs. 9.50%, and 7.00% vs. 4.38%, both P<0.05), whilst the frequencies of DRB1*04 and DRB1*12 alleles were significantly lower (7.13% vs. 10.75%, and 8.63% vs. 14.38%, both P<0.05). For females from the URSA group, the frequency of DRB1*09 allele (14.00%) was significantly higher compared with the controls (9.25%) (P=0.036), whilst the frequency of DRB1*12(8.50%) allele was significantly lower (14.00%) (P=0.014). For males in the URSA group, the frequencies of DRB1*09 and DRB1*13 alleles were significantly higher than those of the controls (15.00% vs. 9.75%, and 9.25% vs. 4.00%, both P<0.05), whilst the frequencies of DRB1*04 and DRB1*12 alleles were significantly lower (5.75% vs. 12.25%, and 8.75% vs. 14.75%, P<0.05).</p><p><b>CONCLUSION</b>The DRB1*09 and DRB1*13 alleles may contribute to the susceptibility of URSA, while DRB1*04 and DRB1*12 alleles may confer a protective effect factors. For females, however, no significant association of DRB1*13 and DRB1*04 alleles with URSA was found.</p>

Association of methionine synthase reductase gene polymorphism with unexplained recurrent spontaneous abortion / 中华妇产科杂志

Objective To explore the relationship between the polymorphism of methionine synthase reductase(MTRR) A66G and the susceptibility to unexplained repeated spontaneous abortion (URSA).Methods Total of 200 Henan Han couples with URSA (URSA group) and 76 Henan Han healthy couples without URSA (control group)were enrolled in this study.Their MTRR A66G genotypes were determined by PCR restriction fragment length polymorphism (PCR-RFLP).Results (1) The allele frequencies of MTRR A66G:the frequencies of allele A and allele G in URSA group were 76.5％ (153/200)in husband and 72.8％ (146/200) in wife,23.5％ (47/200) in husband and 27.2％ (54/200) in wife,respectively.The frequencies of allele A and allele G in control group were 78.9％ (60/76) in husband and 78.3％ (59/76) in wife,21.1％ (16/76) in husband and 21.7％ (16/76) in wife,respectively.The frequencies of allele A and allele G were not significantly different between female and male subjects within the same experimental group (P ＞ 0.05),and also there were not significantly different between the same gender subjects at URAS and control groups(P ＞ 0.05).(2) The genotype frequencies of MTRR A66G:the frequencies of genotype AA,AG and GG in URSA group were 57.0％ (114/200) in husband and 52.0％ (104/200) in wife,39.0％ (78/200) in husband and 41.5％ (83/200) in wife,4.0％ (8/200) in husband and 6.5％ (13/200) in wife,prepectively.The frequencies of genotype AA,AG and GG in control group were 59.2％ (45/76) in husband and 59.2％ (50/76) in wife,39.5％ (30/76) in husband and 38.2％ (29/76) in wife;1.3 ％ (1/76) in husband and 2.6％ (2/76) in wife,prepectively.The frequencies of genotype AA,AG and GG were not significantly different between female and male subjects within the same group (P ＞ 0.05),and also there were not significantly different between the same gender subjects at URSA and control groups (P ＞0.05).(3) Combined genotype of couples:the combined genotype frequencies of GG + GG,GG + AG,GG +AA,AG + AG,AG + AA and AA + AA in URSA group were 1.0％ (2/200),2.5％ (5/200),6.0％ (12/200),20.0％ (40/200),38.0％ (76/200),and 32.5 ％ (65/200),prepectively ; the combined genotype frequencies in control group were 0,1.3％ (1/76),2.6％ (2/76),17.1％ (13/76),42.1％ (32/76),36.8％ (28/76),prepectively.The combined genotype analysis between the two groups were also not significantly different (P ＞ 0.05).Conclusion The polymorphism of MTRR A66G gene was not associated with the susceptibility to URSA (P ＞ 0.05),and so it was not the inherited genetic risk factor of URSA.

The neuroprotective effect of hyperbaric oxygenation on the mitochondria of dopaminergic neurons in a mouse model of Parkinson's disease / 中华物理医学与康复杂志

Objective To investigate any neuroprotective effect of hyperbaric oxygenation (HBO) on the mitochondria of dopaminergic neurons using a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mouse model of Parkinson's disease.Methods Forty-eight male SPF C57BL/6 mice were randomly assigned to either a normal control group (treated with a 30 ml/kg intraperitoneal injection of physiological saline once a day),a model group ( treated with a 30 mg/kg intraperitoneal injection of MPTP once a day) or an HBO therapy group ( treated with a 30 mg/kg intraperitoneal injection of MPTP and HBO once a day).The expression of tyrosine hydroxylase (TH) protein,PINK1 protein and caspase-3 in brain tissue was measured using immunohistochemistry and Western blotting assays.Results Compared with the control group,the expression levels of TH protein and PINK1 protein were significantly lower in the neurons of the substantia nigra in the model mice.HBO therapy upregulated the expression of TH and PINK1 protein.Compared with the control group,the average level of caspase-3 protein in the neurons of the substantia nigra in the model mice was significantly higher.HBO therapy downregulates the expression of caspase-3 protein.Conclusions HBO can protect mitochondria and inhibit apoptosis of dopaminergic neurons in the substantia nigra of brains with (MPTP-induced) Parkinson's disease by upregulating the expression of PINK1 protein and TH protein,and downregulating the expression of caspase-3 protein.