OBJECTIVETo evaluate the feasibility of genetic and prenatal diagnosis for a family affected with pyruvate kinase deficiency (PKD).

METHODSTargeted sequence capture and high-throughput sequencing technology was used to detect the exons and exon-intron boundaries of the PKLR gene in a clinically suspected PKD patient. Meanwhile, the genotype of the pedigree was validated by Sanger sequencing. Prenatal genetic diagnosis was performed by amniotic fluid sampling after genotype of the mother of the proband was determined.

RESULTSThe proband was found to harbor double heterozygous mutations, c.661G>A (Asp221Asn) and c.1528C>T (Arg510Ter), which resulted in amino acid substitution Asp221Asn and Arg510Ter. Such mutations were confirmed by Sanger sequencing. The mother and father of the proband were detected to have respectively carried c.1528C>T (Arg510Ter) and c.661G>A (Asp221Asn) mutation. The fetus was found to have carried the same mutations as the proband. Following selected abortion, analysis of fetal tissue was consistent with the result of prenatal diagnosis.

CONCLUSIONThe compound mutations of c.661G>A and c.1528C>T of PKLR gene probably underlie the PKD in the family. Prenatal diagnosis of the mutations analysis can facilitate detection of affected fetus in time.

Adult ; Anemia, Hemolytic, Congenital Nonspherocytic ; embryology ; enzymology ; genetics ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Pyruvate Kinase ; deficiency ; genetics ; metabolism ; Pyruvate Metabolism, Inborn Errors ; embryology ; enzymology ; genetics

OBJECTIVETo investigate the gene expression profiles in lung adenocarcinoma (LA), tumor adjacent tissue (TAT) and fetal lung tissue (FLT) by cDNA microarray technique.

METHODSTotal RNA from LA, TAT and FLT was extracted and purified. The cDNA was made by RT-PCR, and then labeled with Cy5 and Cy3 fluorescence as probes which were hybridized with the whole gene chips. Subsequently, the signal images were scanned by ScanArray 4000 fluorescence scanner and analyzed by Gene Pix PRO3.0.

RESULTSIn 4 cases with LA and TAT, 25 genes were screened out for differences in gene expression level, among which 3 were upregulated and 22 downregulated; in FLT and TAT cases, 316 genes were screened out, among which 192 were upregulated and 124 downregulated; 16 genes were found to be differentially expressed genes in common in LA, TAT and FLT, among which 12 were upregulated and 4 downregulated.

CONCLUSIONThe 25 differentially expressed genes in LA and TAT may be related to occurrence and development of lung cancer, while the 316 genes in FLT and TAT may be related to fetal developmental. The 16 differentially expressed genes may be related to the initiation of lung cancer.

Adenocarcinoma ; genetics ; metabolism ; Fetal Development ; genetics ; Fetus ; Gene Expression Profiling ; Humans ; Interleukin-6 ; metabolism ; Lung ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis ; Oncogenes ; genetics ; Pyruvate Kinase ; metabolism ; Receptors, CXCR4 ; metabolism ; Thioredoxin-Disulfide Reductase ; metabolism

BACKGROUND/AIMS: M2 pyruvate kinase (M2-PK) is an enzyme that is produced in undifferentiated and proliferating tissues. This study aims to evaluate the usefulness of the immunochromatographic M2 pyruvate kinase (iM2-PK) for the screening of colorectal cancer (CRC) and premalignant lesions. METHODS: Healthy volunteers and patients with colorectal neoplasia were enrolled in six academic hospitals in the capital province of Korea. The iM2-PK value was compared with the immunochromatographic fecal occult blood test (iFOBT) and fecal tumor M2-PK enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 323 subjects were enrolled. The sensitivity of iM2-PK for CRC was 92.8%, which was superior to iFOBT (47.5%, p<0.0001). For adenomatous lesions, the sensitivity of iM2-PK was 69.4%, which was also superior to iFOBT (12.1%, p<0.001). Compared with M2-PK ELISA, iM2-PK exhibited significantly enhanced sensitivity for CRC (97.5% vs 80.0%, p=0.0289). The sensitivity of iM2-PK was higher in advanced stages of CRC compared with cancers confined to the mucosa and submucosa (p<0.05). However, lymph node metastasis had no influence on the sensitivity of iM2-PK. CONCLUSIONS: The iM2-PK exhibited increased sensitivity for identifying CRC and adenomatous lesions compared with iFOBT. Given its rapid results and convenience, CRC screening using iM2-PK is promising.
Adenoma/*diagnosis ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor/*analysis ; Clinical Enzyme Tests/*instrumentation ; Colorectal Neoplasms/*diagnosis ; Enzyme-Linked Immunosorbent Assay ; Feces/*enzymology ; Female ; Healthy Volunteers ; Humans ; Immunochromatography/methods ; Male ; Middle Aged ; Occult Blood ; Precancerous Conditions/diagnosis/enzymology ; Predictive Value of Tests ; Pyruvate Kinase/*analysis ; Reagent Kits, Diagnostic ; Republic of Korea ; Sensitivity and Specificity

OBJECTIVETo characterize the biologic featrues of hepatic oval cells and their protein expression profiles during induced differentiation in vitro.

METHODSRat hepatic oval cells were treated with epidermal growth factor (EGF) and hepatocyte growth factor (HGF) in vitro, followed by morphological and molecular marker assessment by electromicroscopy, immunocytochemistry, RT-PCR and protein expression chip technology.

RESULTSTen weeks after induction, the levels of GST-P mRNA and M2-PK mRNA were significantly reduced, whereas those of ALB and CK18 were elevated. Significant variations of expression was seen in 8 protein species during the course of the induced differentiation.

CONCLUSIONCombined EGF and HGF treatment in vitro induces cell differentiation of hepatic oval cells, a process in which 8 protein species may play some regulatory roles.

Albumins ; metabolism ; Animals ; Cell Differentiation ; drug effects ; Epidermal Growth Factor ; pharmacology ; Glutathione Transferase ; biosynthesis ; genetics ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; metabolism ; ultrastructure ; Immunohistochemistry ; Keratin-18 ; metabolism ; Protein Array Analysis ; Pyruvate Kinase ; biosynthesis ; genetics ; RNA, Messenger ; metabolism ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the difference of genes expression profiles between focal cerebral ischemia tissue and that treated with Baicalin using cDNA microarray.

METHODThe total RNAs were isolated from rat brains of sham-operation, vehicle (focal cerebral ischemia of rat brain) and baicalin-treated groups. mRNAs were reversely transcribed to cDNA with incorporation of fluorescent dUTP (Cy5 or Cy3 dUTP) to prepare hybridization probes. The PCR products of 4096 genes were spotted on the chip after a serial of treatment. The mixed probes were hybridized to the cDNA microarray. Axon Genepix 4000B and GenePixPro 3.0 software were used to scan and analyze the fluorescent signals.

RESULTThe expressions of 199 and 12 genes were found up-regulated and down-regulated, respectively, in the vehicle group compared with the sham-operation one. But the numbers of genes whose expressions were up-regulated and down-regulated were 89 and 88, respectively, when comparing the gene expression in the Baicalin-treated rat brain with that in the vehicle group. Moreover, one down-regulated and three up-regulated genes in the vehicle group were up-regulated and down-regulated in the Baicalin-treated group, respectively. Expressions of three up-regulated genes in the vehicle group were further reinforced in the Baicalin-treatment group.

CONCLUSIONMultiple pathways and nodes may be involved in the pharmacological effect of Baicalin on brain ischemia.

Animals ; Brain Ischemia ; genetics ; metabolism ; Flavonoids ; isolation & purification ; pharmacology ; GTP-Binding Proteins ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; drug effects ; Male ; Oligonucleotide Array Sequence Analysis ; Plants, Medicinal ; chemistry ; Pyruvate Kinase ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Scutellaria ; chemistry ; Vimentin ; genetics ; metabolism