Objective To identify the ABCD1 gene mutation in a patient suspected with adrenoleukodystrophy (ALD) and perform its gene analysis in his family members to make a definite diagnosis. Methods Total RNA and genomic DNA were extracted from the leukocytes of peripheral blood in the proband and the family members. The ABCD1 coding region of cDNA in the proband was amplified and sequenced. Mutational site in the ABCD1 gene of the proband was further confirmed by PGR and direct sequencing; at the same time, the mutation in the ABCD1 gene of the genomic DNA in the family members was analyzed by direct sequencing. Results Two bases deletions (656_657delGA)were identified and the corresponding mutation of fs R89 was detected in the ABCD1 gene of the proband, which could make the definite diagnosis of ALD that belonged to adrenomyeloneuropathy. The same gene mutation (ALD hemizygote) was noted in his cousin; his mother, younger sister of his mother and his younger female cousin were noted as the ALD carrers. His older sister was noted as ABCD1 normal genotype. Conclusions A novel ABCD1 gene mutation (fs R89) was identified in Chinese patient with ALD. Molecular testing is an effective way in making diagnosis on patient suspected as having X-linked ALD.

Adult ; Female ; Hepatitis B, Chronic ; complications ; physiopathology ; Humans ; Liver ; physiopathology ; Liver Function Tests ; Male ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; complications ; physiopathology

OBJECTIVECoronary heart disease (CHD) is one of the most common causes of death in the world. Some studies suggested that CHD begins in childhood. Obesity and dyslipidemia are important risk factors of coronary heart disease. Apolipoprotein (apo)E gene associated with dyslipidemia and coronary heart disease. The present study was designed to investigate the expression status of apoE gene in peripheral blood monocyte and association of apoE gene expression with lipids in children with obesity.

METHODSAmong 32 children with obesity and 32 healthy children without obesity or overweight, ApoE gene expressions were determined by competitive reverse transcription-polymerase chain reaction in peripheral blood monocyte. The concentrations of plasma triglyceride, total cholesterol, low density lipoprotein-cholesterol, high density lipoprotein-cholesterol, lipoprotein(a), apoA I, apoB(100) and apoE were measured.

RESULTSExpression of apoE gene was detected in peripheral blood monocyte. Expression of apoE gene was significantly reduced in children with obesity as compared with control group (0.29 +/- 0.14 moles/mole GAPDH mRNA vs. 0.36 +/- 0.10 moles/mole GAPDH mRNA, t = 2.15, P < 0.05). The more severe was the degree of obesity, the more significantly reduced the expression of apoE gene; the degree of obesity was negatively correlated with the levels of expression of apoE gene (correlation coefficient = -0.40, P < 0.05). Compared with control group, the levels of triglyceride, total cholesterol, low density lipoprotein-cholesterol, and apoB(100) were higher, and those of high density lipoprotein-cholesterol, apoA I and apoE were lower in children with obesity [(1.68 +/- 0.50) mmol/L vs. (0.99 +/- 0.54) mmol/L, (4.47 +/- 0.91) mmol/L vs. (3.33 +/- 0.90) mmol/L, (2.23 +/- 0.71) mmol/L vs. (1.13 +/- 0.96) mmol/L, (94.48 +/- 9.97) mg/dl vs. (83.81 +/- 15.64) mg/dl, (1.47 +/- 0.39) mmol/L vs. (1.73 +/- 0.36) mmol/L, (112.71 +/- 27.86) mg/dl vs. (134.80 +/- 45.36) mg/dl, (24.50 +/- 10.92) mg/L vs.(35.07 +/- 9.79) mg/L, respectively, P < 0.05]. ApoE gene expression was associated with plasma lipids metabolism in children with obesity. The quantity of apoE gene expression was inversely associated with low density lipoprotein-cholesterol, positively correlated with apoE (correlation coefficient = -0.33, 0.35, respectively, P < 0.05). The quantity of apoE gene expression was not associated with total cholesterol, triglyceride, high density lipoprotein-cholesterol, lipoprotein(a), apoA I, and apoB(100) (correlation coefficient = -0.19, -0.11, 0.16, 0.09, 0.18, 0.22, P > 0.05).

CONCLUSIONExpression of apoE gene was significantly reduced in peripheral blood monocyte in children with obesity. The quantity of apoE gene expression was associated with degree of obesity and abnormality of blood lipids.

Apolipoproteins E ; genetics ; Child ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene Expression ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Obesity ; blood ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Triglycerides ; blood

Objective To explore the effect and molecular mechanism of miR-202 on the differentiation of 3T3-L1 preadipocyte .Methods Through lentivirus infected with 3T3-L1 preadipocytes , we set up the AMO-miR-202 group and the random control group , then, these cells were induced to differentiate , nine days later, differentiation was assessed by Oil Red O staining and we examined the mRNA expression of PPARγ2 and aP2 by RT-PCR method. We examined the mRNA expression of PPARγ2,aP2 and PGC1βby Western blot method .Results After packa-ging lentivirus with AMO-miR-202 and random sequence control miRNA through cell line 293T, 80%-90%cells with fluorescence were found under fluorescence microscope; After these two lentivirus respectively infected with 3T3-L1 preadipocytes, About 70%-80%cells with fluorescence were found under fluorescence microscope .Oil Red O staining test showed that these cells with Oil Red O stained bright red fat droplets of AMO-miR-202 group and PPARγ2 and aP2 mRNA expression in the AMO-miR-202 group significantly lower than control groups (P<0.05). Western blot assay showed that the protein expression of PGC 1βin the AMO-miRNA-202 group was significantly increased(P<0.05), but the expression of aP 2 and PPARγ2 was significantly decreased (P<0.01).However, the random control group and the adipocyte group had no significant effect on the above indexes .Conclusions miR-202 can promote the differentiation of 3T3-L1 preadipocyte by inhibiting the protein expression of PGC 1βand im-proving the protein expression of PPARγ2 and aP2.