ABSTRACT
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.
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Objective To study the effectiveness of the laboratory diagnosis of multiple myeloma(MM)patients with immun-ofixtion electrophoresis (IFE),protein electrophoresis (SPE)and immunoglobulins and light chain quantitative analysis. Methods Selected 192 MM patients and 30 healthy controls during June 2012 to December 2013 and analyzed the results of IFE,SPE and immunoglobulins,and light chain quantitative analysis in MM patients.Results M protein bands were seen in 120 cases (62.5%)by using SPE and M protein were positive in 174 cases (90.6%)among the 192 MM patients by using IFE.IFE showed that IgG were maximum type of the M protein (106 of 192,55.2%).There were IgG-λ type 66 cases (34.4%),IgA type 36 cases (18.8%)and free light chain type 24 cases (12.5%).Immunoglobulins of different immuno-phenotypes had higher than the nomal group with serum immunoglobulin and light chain quantitative analysis (P <0.05). The detection rate was 67.7% (130/192).Whateverκ-type M protein orλ-type M protein,the ratio ofκ/λwas significantly abnormal (P <0.05).The detection rate was 85.4% (164/192).Conclusion The better detection rate of immunological techniques such as immunofixtion electrophoresis and immunoglobulins quantitative analysis might provide valuable basis for the diagnosis and treatment of MM clinically and prevent misdiagnosis.
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Objective To study the protective effects of DADLE on pulmonary function in rats with sepsis and the mechanism.Methods Seventy-two SD rats were randomly divided into sham operated group(SO),septic group(SEP),and DADLEtreated group (DADLE).Sepsis model was induced by cecal ligation and puncture(CLP).In SO group,the abdomen was opened without any other treatment.In DADLE-treated group,DADLE(0.5 mg/ml) was administerd at a dose of 5 ml/kg by intravenous injection after CLP.Rats were sacrificed at the 2nd,6th and 10th h after CLP.Arterial blood was collected for blood gas analysis.The wet-to-dry lung weight ratio was measured,and the levels of TNF-a and IL-6 in blood were detected.The levels of MPO,MDA and ATP in lung tissue were determined.The pathologic changes of the lungs were examined under a light microscope.Results As compared with SEP group,PaCO_2,and PaO_2 were significantly increased(P<0.05),while the wet-to-dry ratio and levels of serum TNF-a and IL-6 were significantly reduced(P<0.05),and levels of MPO and MDA were significantly reduced(P<0.05) and content of ATP in lung tissue was significantly increased (P<0.05)in DADLE group.More severe pathological changes were found in SEP group than in DADLE group.Conclusion DADLE has pulmonary protective effects in rats with sepsis.