OBJECTIVETo perform cytogenetic analysis for children, especially newborns suspected for chromosome abnormalities.

METHODSPeripheral blood or born marrow specimens were respectively cultured in proper media. Karyograms were analyzed following G-banding.

RESULTSOf 154 blood specimens, numerical chromosomal abnormalities were identified in 20 patients, which included 19 with trisomy 21. Structural aberrations were identified in 13 patients, among which chromosome 9 aberrations were seen in 6 cases. These included 3 inversions, 1 deletion, 1 insertion and 1 duplication. All aberrations were located in pericentromere region of chromosome 9 with clinical manifestations including congenital heart disease, peculiar facial appearance, paralysis, dysplasia and/or movement disorder. Chromosome polymorphisms were found in 20 patients, most of which had absence of satellites or variation of heterochromatin on chromosome 9. Of 10 bone marrow specimens from children suspected for acute leukemia, chromosome abnormalities were identified in 5 patients.

CONCLUSIONCytogenetic analysis is useful for children featuring multiple congenital abnormalities. Chromosome 9 abnormalities and their clinical relevance should attract more attention.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; Chromosome Banding ; Chromosome Disorders ; epidemiology ; Chromosomes, Human, Pair 9 ; Humans ; Infant ; Infant, Newborn ; Physical Chromosome Mapping ; Prevalence

Chromosome Mapping ; Enhancer Elements, Genetic ; Erythropoietin ; genetics ; Exercise ; Humans ; Military Personnel ; Physical Endurance ; Polymorphism, Single Nucleotide ; Young Adult

The incidence of type 2 diabetes is rising rapidly because of an increase in the incidence of being overweight and obesity. Identification of genetic determinants for complex diseases, such as type 2 diabetes, may provide insight into disease pathogenesis. The aim of the study was to investigate the shared genetic factors that predispose individuals to being overweight and developing type 2 diabetes. We conducted genome-wide linkage analyses for type 2 diabetes in 386 affected individuals (269 sibpairs) from 171 Korean families and association analyses with single-nucleotide polymorphisms of candidate genes within linkage regions to identify genetic variants that predispose individuals to being overweight and developing type 2 diabetes. Through fine-mapping analysis of chromosome 4q34-35, we detected a locus potentially linked (nonparametric linkage 2.81, logarithm of odds 2.27, P=6 x 10-4) to type 2 diabetes in overweight or obese individuals (body mass index, BMI> or =23 kg m-2). Multiple regression analysis with type 2 diabetes-related phenotypes revealed a significant association (false discovery rate (FDR) P=0.006 for rs13144140; FDR P=0.002 for rs6830266) between GPM6A (rs13144140) and BMI and waist-hip ratio, and between NEIL3 (rs6830266) and insulin level from 1314 normal individuals. Our systematic search of genome-wide linkage and association studies, demonstrate that a linkage peak for type 2 diabetes on chromosome 4q34-35 contains two type 2 diabetes-related genes, GPM6A and NEIL3.
Body Mass Index ; Chromosomes, Human, Pair 4/*genetics ; Diabetes Mellitus, Type 2/*complications/*genetics ; Female ; Genetic Linkage ; *Genetic Loci ; *Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Male ; Middle Aged ; Overweight/*complications/*genetics ; Phenotype ; Physical Chromosome Mapping ; Statistics, Nonparametric

We report a long-term follow-up of unstable hemoglobin (Hb) patient. He was diagnosed as Hb Madrid [beta115(G17)Ala-->Pro] by direct DNA sequencing and restriction enzyme analysis. Hydroxyurea had been given for beta-chain hemoglobinopathies through activation of gamma(gamma)-chain synthesis. Nowadays he still needs transfusion three or four times per year, but he had been free of hemolytic crisis after hydroxyurea. Although he has been treated for hemochromatosis with parenteral and oral iron-chelating agents, liver cirrhosis complicated by esophageal varix was developed and treated with endoscopic ligation. In addition, he is on warfarin maintenance for anticoagulation therapy for extensive portal vein and superior mesenteric vein thrombosis which presented with abdominal pain and diagnosed by CT scan. In management of unstable Hb patients, physician should monitor and control the serum ferritin level with iron-chelating agents, and be aware of possible long-term complication including hemochromatosis, cirrhosis or thromboembolism.
Abdominal Pain ; Esophageal and Gastric Varices ; Ferritins ; Fibrosis ; Follow-Up Studies ; Hemochromatosis ; Hemoglobinopathies ; Hemoglobins ; Hemoglobins, Abnormal ; Humans ; Hydroxyurea ; Ligation ; Liver Cirrhosis ; Mesenteric Veins ; Organothiophosphorus Compounds ; Portal Vein ; Restriction Mapping ; Sequence Analysis, DNA ; Thromboembolism ; Thrombosis ; Warfarin

PURPOSE: We applied a simplified method using polymerase chain reaction (PCR)-based enzymatic digestion for the detection of epidermal growth factor receptor (EGFR) mutation. MATERIALS AND METHODS: We selected 74 samples of adenocarcinoma of the lung with EGFR exons 19 and 21 that had been previously sequenced. We designed PCR primers and chose a DNA restriction enzyme. Seventy four additional lung cancer samples were tested as a test set. For test sets, the PCR-based method was performed first, followed by validation of the result by DNA sequencing. RESULTS: In the first sample group, we found 15 (20.3%) mutations in exon 19, and 9 (12.2%) mutations in exon 21 using the sequencing method. By using the PCR-based method, we were able to identify all of the mutated samples detected by the sequencing method. The PCR-based method also detected mutations in exon 19 in three additional samples and in exon 21 in one additional sample. In the second sample group, by performing the PCR-based method, we found 10 (13.5%) and 7 (9.5%) mutations in exons 19 and 21, respectively. Additional mutations in exon 19 were identified in 2 samples by the sequencing method. However, the sequencing method failed to identify a mutation in exon 21 in one sample. CONCLUSION: The sensitivity of the PCR-based enzymatic digestion method seems to be comparable to that of the traditional sequencing method for detecting EGFR mutations. Our method can be widely used as a screening test to select patients who may benefit from EGFR targeted therapy.
Adenocarcinoma ; Carcinoma, Non-Small-Cell Lung ; Digestion ; DNA ; DNA Restriction Enzymes ; Epidermal Growth Factor ; Exons ; Genes, erbB-1 ; Humans ; Lung ; Lung Neoplasms ; Mass Screening ; Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; Restriction Mapping

Serratia marcescens jn01 was employed as the host for the isolation of phages from environmental sewage. One strain of phage named SmPjn was purified by picking transparent plaque with 2mm diameter and clear edge on the double-layer agar repeatedly. Electron micrographs indicated that the phage head was icosahedral with head size and tail length of (58 +/- 2.16) x (55 +/- 0.47) nm and (7 +/- 1.25) nm, respectively. On the basis of the morphology, this phage belongs to the family Podoviridae. Host-range determination revealed that the phage was capable of infecting the other two isolates of S. marcescens, P25 and CMCC41002. The optimal multiplicity of infection was 1. A one-step growth curve of SmPjn indicated that the latent period and burst size were estimated at 50 min and 1,125 pfu/cell, respectively . Genomic DNA of SmPjn was above 27kb in size and could be digested by Hind Ill and EcoR I into 11 and 9 visible fragments after electrophoresis, respectively. A novel Podoviridae-phage infecting S. marcescens was firstly reported in China.
China ; DNA, Viral ; genetics ; isolation & purification ; metabolism ; Host Specificity ; Podoviridae ; genetics ; growth & development ; isolation & purification ; Restriction Mapping ; Serratia marcescens ; physiology

OBJECTIVETo identify the RUNX2 gene mutation in two unrelated Chinese families with cleidocranial dysplasia (CCD), and to assess the feasibility of gene diagnosis for patients with CCD.

METHODSGenomic DNA was isolated from peripheral blood samples of 4 patients and 4 healthy members in the two pedigrees as well as 102 unrelated healthy controls. All 7 coding exons and their flanking intronic sequences of the RUNX2 gene were amplified by PCR, then the PCR products were sequenced bi-directionally. The sequencing results were compared with normal sequences in GenBank to identify the mutation. The mutation was confirmed by RFLP with restriction endonuclease.

RESULTSIn one family, a novel heterozygous missense mutation c.346T to A (W116R) in exon 1 of the RUNX2 gene was detected in the two affected individuals, and the mutation was further confirmed with Bsr I restriction endonuclease digestion. In the other family, a novel nonsense mutation c.610A TO T (K204X) was identified in the two patients. No above sequence change was found in the 102 healthy controls.

CONCLUSIONTwo novel RUNX2 mutations were found in two unrelated Chinese families with cleidocranial dysplasia. The identification of these mutations further extended the mutation spectrum of RUNX2 gene and will facilitate prenatal diagnosis and gene diagnosis of CCD.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Cleidocranial Dysplasia ; genetics ; physiopathology ; Core Binding Factor Alpha 1 Subunit ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Infant ; Male ; Mutation ; Pedigree ; Restriction Mapping

The aim of this study was to evaluate the presence of Helicobacter (H.) spp. in swine affected by gastric ulceration. Stomachs from 400 regularly slaughtered swine were subjected to gross pathological examination to evaluate the presence of gastric ulcers. Sixty-five samples collected from ulcerated pars esophagea and 15 samples from non-ulcerated pyloric portions were submitted to histopathological and molecular analyses, to detect Helicobacter spp., H. suis and H. pylori by PCR. Feces and saliva swabs were also collected from 25 animals in order to detect in vivo the presence of Helicobacter spp.. Gastric ulcers were detected in 373 cases (93%). The presence of ulcers in association with inflammatory processes was further confirmed by histological examination. Forty-nine percent (32/65) of the ulcerated esophageal portions as well as 53% (8/15) of the non-ulcerated pyloric portions were positive for Helicobacter spp. by PCR. The Helicobacter spp. positive samples were also positive for H. suis, while H. pylori was not detected. These results were confirmed by restriction enzyme analysis. With regard to feces and saliva samples, 15/25 (60%) and 16/25 (64%) were positive for Helicobacter spp. PCR, respectively but all were negative in H. suis and H. pylori specific PCR.
Animals ; Feces/*microbiology ; Helicobacter/*isolation & purification ; Polymerase Chain Reaction/veterinary ; Restriction Mapping/veterinary ; Saliva/*microbiology ; Stomach/*microbiology ; Stomach Ulcer/microbiology/pathology/*veterinary ; Swine ; Swine Diseases/*microbiology/pathology

BACKGROUND: Extended-spectrum cephalosporins (ESCs)-resistant Enterobacter cloacae is one of the pathogens of nosocomial infection the incidence of which is on the rise. Moreover, plasmid-mediated quinolone resistance genes (PMQR genes) that reduce quinolone sensitivity have been shown to be widely distributed among clinical isolates of Enterobacteriaceae. This study was carried out to observe the distribution of PMQR genes in clinical isolates of E. cloacae resistant to ESCs. MATERIALS AND METHODS: Fourty-three ESCs-resistant E. cloacae strains from the blood collected during the span of 7 years, from 1994 to 2001, at Seoul National University Children's Hospital (SNUCH) were included in this study. Isoelectric focusing and enzyme specific PCR were performed to characterize beta-lactamase. The presence of qnrA, qnrB, qnrC, qnrS, qepA, and aac(6')-Ib-cr was determined by PCR, restriction enzyme analyses of PCR products, and DNA sequencing. Minimal inhibitory concentration (MIC) of several quinolones was measured by agar dilution method. RESULTS: The PMQR genes were detected in 9 (21%) of 43 ESCs-resistant E. cloacae isolates. Among them, five isolates were positive for qnrB2, and each two isolates harbored qnrB4 or qnrB5, respectively. qnrA, qnrC, qnrS or qepA was not identified. aac(6')-Ib was detected in 27 isolates, but aac(6')-Ib-cr was not found. Among the 9 qnrB-positive isolates, 5 produced SHV-12, 3 were derepressed mutants, and 1 produced pI 7.5 beta-lactamase. MIC ranges and percent resistances of nalidixic acid, ofloxacin, levofloxacin, and moxifloxacin for the PMQR genes-positive isolates were higher than PMQR genes-negative isolates. CONCLUSION: In this study, ESCs-resistant E. cloacae showed a high prevalence of PMQR genes, and qnrB was the only PMQR gene identified.
Agar ; Aza Compounds ; beta-Lactamases ; Cephalosporins ; Cloaca ; Cross Infection ; Enterobacter ; Enterobacter cloacae ; Enterobacteriaceae ; Incidence ; Isoelectric Focusing ; Nalidixic Acid ; Ofloxacin ; Polymerase Chain Reaction ; Prevalence ; Quinolines ; Quinolones ; Restriction Mapping ; Sequence Analysis, DNA

BACKGROUND: Extended-spectrum cephalosporins (ESCs)-resistant Enterobacter cloacae is one of the pathogens of nosocomial infection the incidence of which is on the rise. Moreover, plasmid-mediated quinolone resistance genes (PMQR genes) that reduce quinolone sensitivity have been shown to be widely distributed among clinical isolates of Enterobacteriaceae. This study was carried out to observe the distribution of PMQR genes in clinical isolates of E. cloacae resistant to ESCs. MATERIALS AND METHODS: Fourty-three ESCs-resistant E. cloacae strains from the blood collected during the span of 7 years, from 1994 to 2001, at Seoul National University Children's Hospital (SNUCH) were included in this study. Isoelectric focusing and enzyme specific PCR were performed to characterize beta-lactamase. The presence of qnrA, qnrB, qnrC, qnrS, qepA, and aac(6')-Ib-cr was determined by PCR, restriction enzyme analyses of PCR products, and DNA sequencing. Minimal inhibitory concentration (MIC) of several quinolones was measured by agar dilution method. RESULTS: The PMQR genes were detected in 9 (21%) of 43 ESCs-resistant E. cloacae isolates. Among them, five isolates were positive for qnrB2, and each two isolates harbored qnrB4 or qnrB5, respectively. qnrA, qnrC, qnrS or qepA was not identified. aac(6')-Ib was detected in 27 isolates, but aac(6')-Ib-cr was not found. Among the 9 qnrB-positive isolates, 5 produced SHV-12, 3 were derepressed mutants, and 1 produced pI 7.5 beta-lactamase. MIC ranges and percent resistances of nalidixic acid, ofloxacin, levofloxacin, and moxifloxacin for the PMQR genes-positive isolates were higher than PMQR genes-negative isolates. CONCLUSION: In this study, ESCs-resistant E. cloacae showed a high prevalence of PMQR genes, and qnrB was the only PMQR gene identified.
Agar ; Aza Compounds ; beta-Lactamases ; Cephalosporins ; Cloaca ; Cross Infection ; Enterobacter ; Enterobacter cloacae ; Enterobacteriaceae ; Incidence ; Isoelectric Focusing ; Nalidixic Acid ; Ofloxacin ; Polymerase Chain Reaction ; Prevalence ; Quinolines ; Quinolones ; Restriction Mapping ; Sequence Analysis, DNA