The aim of this study was to investigate the clinical features and DGUOK gene mutations of an infant with mitochondrial DNA depletion syndrome (MDS). The patient (more than 7 months old) manifested as hepatosplenomegaly, abnormal liver function, nystagmus and psychomotor retardation. Genetic DNA was extracted from peripheral blood samples of the patient and her parents. Targeted Exome Sequencing was performed to explore the genetic causes. Sanger sequencing was carried out to confirm the detected mutations. The sequencing results showed that the patient was a compound heterozygote for c.679G>A and c.817delT in the DGUOK gene. The former was a reportedly pathogenic missense mutation of maternal origin, while the latter, a frameshift mutation from the father, has not been described yet. The findings in this study expand the mutation spectrum of DGUOK gene, and provide molecular evidence for the etiologic diagnosis of the patient as well as for the genetic counseling and prenatal diagnosis in the family.
Female ; Humans ; Infant ; Mitochondrial Diseases ; genetics ; therapy ; Mutation ; Phosphotransferases (Alcohol Group Acceptor) ; chemistry ; genetics

OBJECTIVEThis research intends to amplify the entire coding region sequences of SLC25A13 mRNA which encodes citrin, and to investigate sequence features of the transcripts for this gene in cultured human amniocytes. This study will provide laboratory evidence for prenatal diagnosis of neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) at mRNA level.

METHODSOne amniocyte sample was collected from a pregnant woman who underwent prenatal diagnosis of citrin deficiency and whose fetus has proven a carrier of 851del4 mutation by genomic DNA analysis. Another amniocyte sample, as a control, was from a fetus without family history of citrin deficiency. Total RNA was extracted from cultured amniocytes, cDNA was synthesized, and then nested-PCR was performed to amplify the entire coding region sequences of SLC25A13. The PCR products were cloned and analyzed by sequencing.

RESULTSThe entire coding region of SLC25A13 gene was successful amplified from two cultured human amniocytes. The splice variant of SLC25A13, SLCA (normal mRNA), was identified in the two samples. SLCB (CAG insertion between exon 9-10) was identified in the control. SLCC (exon 5-11 skipping), but not transcriptional product from the allele with 851del4 mutation, was identified in the 851del4 mutation carrier.

CONCLUSIONSThis study demonstrated that the entire coding region of SLC25A13 cDNA can be successfully amplified from two cultured human amniocytes, and revealed exon 5-11 skipping as a novel SLC25A13 transcript. Normal mRNA predominated in the transcripts in normal control and 851del4 mutation carrier, suggesting that the two fetuses were not at risk for NICCD. These SLC25A13 transcription features provided laboratory evidence for prenatal diagnosis of NICCD.

Amniotic Fluid ; cytology ; metabolism ; Calcium-Binding Proteins ; deficiency ; Cholestasis, Intrahepatic ; diagnosis ; Cloning, Molecular ; Female ; Humans ; Mitochondrial Membrane Transport Proteins ; genetics ; Organic Anion Transporters ; deficiency ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis ; methods ; RNA, Messenger ; analysis ; Sequence Analysis, DNA ; Transcription, Genetic

Animals ; Cell Proliferation ; Hepatic Stellate Cells ; Liver Cirrhosis/metabolism* ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; Rats

OBJECTIVETo investigate the expression differences of apoptosis-inducing factor (AIF) and caspase-3 among colorectal carcinoma, adenoma and normal mucosa, and to identify the relationship between AIF and caspase-3 expression in colorectal adenoma-carcinoma sequence.

METHODSFormalin-fixed paraffin embedded colorectal tissues from 174 cases, including 84 adenomas, 72 carcinomas, and 18 normal mucosa, were examined for expression of AIF and caspase-3 by streptavidin-peroxidase (SP) immunohistochemistry.

RESULTSThe positive rates of AIF and caspase-3 in colorectal adenoma were higher than those in normal mucosa (P <0.05). The positive rate of AIF in adenoma showed no significant difference compared to colorectal carcinoma (P >0.05). However, caspase-3 expression in adenomas was significantly higher than that in carcinoma (P <0.05). The positive rate of AIF in tubular adenoma was significantly higher than that in villous adenoma (P <0.05), while the positive expression rate of caspase-3 in the two types of adenoma showed no significant difference (P >0.05). AIF expression had no prominent correlation with the caspase-3 expression (P >0.05).

CONCLUSIONSThe dysregulation of caspase-independent pathway of apoptosis may be an early event in the development of colorectal carcinogenesis, while the dysregulation of the caspase-dependent pathway of apoptosis may be one of contributing factors of colorectal carcinogenesis. The caspase-independent pathway of apoptosis and the caspase-dependent pathway of apoptosis are two relatively independent pathways in colorectal carcinogenesis.

Adenoma ; metabolism ; pathology ; Adult ; Aged ; Apoptosis Inducing Factor ; metabolism ; Caspase 3 ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Intestinal Mucosa ; metabolism ; pathology ; Middle Aged ; Young Adult

Objective To achieve detailed genomic characterization and investigate the antibiotic-resistant mechanisms of plasmid pA1137 carrying the aminoglycoside resistance gene aacC2.Methods Antibiotic-resistant genes were deter-mined by PCR.Conjugation experiments were performed to verify the transferability of plasmid pA 1137.The minimum in-hibitory concentration(MIC)values of bacterial strains were tested with microdilution method.The genetic background, mobile elements and antibiotic resistance mechanisms of pA 1137 were determined using a whole genome sequencing meth-od.Results Both carbapenem-resistant gene blaIMP-8and aminoglycoside-resistant genes aacC2 and aacA4 were carried by A1137 isolated from Enterobacter cloacae(ECL).aacC2 was located in plasmid pA1137 while the other two resistant genes were observed in chromosomes.Plasmid pA1137 was an IncFⅡplasmid,whose total length was 68.97 kb,and GenBank accession number was MF190369.Plasmid pA1137 contained multiple replicons and intact conjugative transfer regions,so it could be transferred into ECL through conjugation experiments and confer corresponding antibiotic resistance to the transconjugant A1137-EC600.Conclusion IncFⅡ plasmid pA1137 has a single accessory region, the first reported Tn5403-based aacC2-tmrB-related region,which can cause stable inheritance and mediate the resistance to aminoglycoside antibiotics in ECL A1137.

This study aimed to report the clinical characteristics and COMP gene mutation of a family with pseudoachondroplasia (PSACH), a relatively rare spinal and epiphyseal dysplasia that is inherited as an autosomal dominant trait. Clinical information on a 5-year-2-month-old PSACH child and his parents was collected and analyzed. Diagnosis was confirmed by PCR amplification and direct sequencing of all the 19 exons and their flanking sequences of COMP gene, and the mutation was further ascertained by cloning analysis of exon 10. The child presented with short and stubby fingers, bow leg, short limb dwarfism and metaphysic broadening in long bone as well as lumbar lordosis. A mutation c.1048_1116del (p.Asn350_Asp372del) in exon 10, inherited from his father who did not demonstrate any phenotypic feature of PSACH, was detected in the child. PSACH was diagnosed definitively by means of COMP mutation analysis, on the basis of the child's clinical and imaging features. The non-penetrance phenomenon of COMP mutation was described for the first time in PSACH.
Achondroplasia ; genetics ; Cartilage Oligomeric Matrix Protein ; genetics ; Child, Preschool ; Cloning, Molecular ; Humans ; Male ; Mutation