OBJECTIVETo investigate EXT1 and EXT2 genes mutations in a family with hereditary multiple osteochondromas (HME).

METHODSA four-generation family with HME from Linyi city of Shandong Province was studied. There were 6 affected individuals among the 17 family members. Physical examination and radiographical evaluations were carried out for all family members. Genomic DNA was extracted from peripheral venous blood and the samples were subjected to mutation screening by PCR of the coding regions of EXT1 and EXT2 genes.

RESULTSThe family has featured an autosomal dominant inheritance pattern. Sequencing of the EXT1 and EXT2 genes suggested the causative gene in this family was in linkage with the second exon of EXT2. A c.244delG mutation was detected, which has resulted in a frameshift mutation p.Asp81IlefsX30. The mutation was found in all of the 6 affected individuals but not in normal family members. And the mutation has co-segregated with the phenotype.

CONCLUSIONThe mutation c.244delG in the EXT2 gene is the probably the cause of the disease in this family.

Adult ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; N-Acetylglucosaminyltransferases ; genetics ; Pedigree ; Point Mutation ; Young Adult

OBJECTIVETo determine the linkage between Smith-Fineman-Myers syndrome (SFMS) and X-linked nuclear protein(XNP) locus.

METHODSPolymerase chain reaction and denaturing polyacrylamide gel electrophoresis were used to genotype two polymorphic short tandem repeats within XNP gene.

RESULTSOne of the two short tandem repeats was informative in SFMS family from Shandong, China. Recombination between SFMS locus and XNP gene was observed in the SFMS family.

CONCLUSIONXNP gene is not associated with the disease in the SFMS family from Shandong, China. SFMS exhibits locus heterogeneity at molecular level.

Abnormalities, Multiple ; genetics ; Craniofacial Abnormalities ; genetics ; DNA Helicases ; Female ; Genetic Linkage ; Growth Disorders ; genetics ; Humans ; Intellectual Disability ; genetics ; Male ; Muscle Hypotonia ; genetics ; Nuclear Proteins ; genetics ; Pedigree ; Phenotype ; Polymorphism, Genetic ; Recombination, Genetic ; Syndrome ; X Chromosome ; X-linked Nuclear Protein

OBJECTIVETo determine the genomic structure of low density lipoprotein receptor related protein 5 (LRP5) gene.

METHODScDNA sequence encoding LRP5 was used to screen genomic clones containing LRP5 gene by computer hybridization approach. By comparing the cDNA sequence of LRP5 with the genomic sequences, the genomic structure of LRP5 was determined, and then it was conformed by amplifying and sequencing the sequences of exons and splicing junction.

RESULTSThe genomic sequence of LRP5 gene was 131.6 kb in length, containing 23 exons and 22 introns. Three single nucleotide polymorphisms were detected within the coding sequences of LRP5 gene, namely A459G in exon 2, C2220T in exon 10 and G4416C in exon 21. Four polymorphic markers, D11S1917, D11S4087, D11S1337 and D11S4178, located in the 5' flank sequence, introns 1, 4, and 13 of the LRP5 gene, respectively.

CONCLUSIONThe characterization of genomic structure of LRP5 gene allows the investigators to detect disease-causing mutation within the gene and further study the function of LRP5 gene.

Base Sequence ; DNA ; chemistry ; genetics ; Exons ; Genes ; genetics ; Humans ; Introns ; LDL-Receptor Related Proteins ; Low Density Lipoprotein Receptor-Related Protein-5 ; Polymorphism, Single Nucleotide ; Receptors, LDL ; genetics ; Sequence Analysis, DNA

OBJECTIVETo study the clinical manifestations and identify causative mutations for a Chinese family affected with X-linked Charcot-Marie-Tooth disease.

METHODSClinical, electrophysiological and pathological features of the family were carefully analyzed by neurologists. Blood samples were obtained from the proband and other family members. Genomic DNA was extracted. Mutation analysis of GJB1 gene was analyzed with PCR and direct sequencing.

RESULTSThe family has fit with X-linked inheritance, and the affected individuals have typical clinical manifestations. A c.614A>G (p.Asn205Ser) mutation was detected in the GJB1 gene in all affected individuals in the family.

CONCLUSIONA c.614A>G (p.Asn205Ser) mutation of GJB1 gene is co-segregated with the disease phenotype in this family and probably underlies the disease.

Asian Continental Ancestry Group ; genetics ; Charcot-Marie-Tooth Disease ; genetics ; Child ; Connexins ; genetics ; Female ; Genes, X-Linked ; genetics ; Genetic Diseases, X-Linked ; genetics ; Humans ; Male ; Mutation ; Pedigree

OBJECTIVETo analyze clinical features and mutation in MYH9 gene for a family featuring autosomal dominant May-Hegglin anomaly.

METHODSClinical and pathological features of all family members were analyzed. Blood samples were collected from the proband and other family members, and genomic DNA was extracted. Potential mutations of MYH9 gene exons 10, 25, 26, 30, 38 and 40 were screened with PCR and direct sequencing. After a mutation was identified in the proband, other affected members as well as healthy members from this family were analyzed with a pair of primers to amplify the mutant site. The PCR products were digested with Taq I enzyme and analyzed with agarose gel electrophoresis.

RESULTSAll affected members had bleeding tendency and typical features including giant platelets, thrombocytopenia and characteristic Dohle body-like leukocyte inclusions. A heterozygous missense mutation c.5521G>A (p.Glu1841Lys) in exon 38 of the MYH9 gene was identified in all affected members from this family.

CONCLUSIONThe variant, c.5521G>A (p.Glu1841Lys) of MYH9, has co-segregated with the phenotype in the family. The mutant site is a hot spot in Chinese population.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Exons ; Female ; Genes, Dominant ; Hearing Loss, Sensorineural ; Humans ; Male ; Molecular Motor Proteins ; genetics ; Mutation ; Myosin Heavy Chains ; genetics ; Pedigree ; Phenotype ; Thrombocytopenia ; diagnosis ; genetics

OBJECTIVETo detect potential mutation in a Chinese family affected with autosomal-dominant synpolydactyly and to provide the basis for prenatal diagnosis.

METHODSInheritance pattern was determined by clinical examination and pedigree analysis. Blood samples were obtained from members of the family. Genomic DNA was extracted and sequenced following PCR amplification. Suspected mutation was confirmed by subclone sequencing and agarose gel electrophoresis.

RESULTSA 27 bp expansion mutation in exon 1 of HOXD13 was identified in all affected individuals from the family but not in unaffected members and normal controls. The mutation has caused insertion of 9 alanines in the polyalanine-expansion region of HOXD13 protein.

CONCLUSIONA polyalanine-expansion within the HOXD13 probably underlies the disease in this family.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA Mutational Analysis ; Exons ; Female ; Genes, Dominant ; Homeodomain Proteins ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Syndactyly ; genetics ; Transcription Factors ; genetics

OBJECTIVE:To establish HPLC fingerprint of Miao medicine Fufukang spray. METHODS:The determination was performed on Thermo Hypersil GOLD C18 column with mobile phase consisted of acetonitrile-0.2％ glacial acetic acid (gradient elution) at the flow rate of 0.6 mL/min. The detection wavelength was set at 262 nm,and column temperature was 25 ℃. The sample size was 10 μL. Using 6′-hydroxyl-justicidin B as reference control,HPLC chromatograms of 10 batches of samples were drawn. Similarity evaluation system software(2004 A edition)of TCM fingerprint was used to evaluate the similarity and determine common peaks of 10 batches of samples. The established method was used to determine the contents of 6′-hydroxyl-justicidin B and justicidin B in 10 batches of samples. RESULTS:There were 12 common peaks in HPLC fingerprints of 10 batches of samples with similarity>0.90. HPLC fingerprint showed a good consistency with the control fingerprint. The contents of 6′-hydroxyl-justicidin B and justicidin B in 10 batches of samples were 1.64-5.84,0.83-2.78 mg/100 mL,respectively. CONCLUSIONS:The established method is simple,accurate,reliable and suitable for quality evaluation of Fufukang spray.

OBJECTIVETo study the prevalence of methylenetetrahydrofolate reductase (MTHFR) C677T genotype and its association with deep vei n thrombophilia in Chinese.

METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was conducted to examine mutation with 63 deep vein thrombophilic patients and 80 health controls in Shandong Hans. The genotype frequencies were calculated by gene counting in patients and controls, and an analysis was made on the association of MTHFR C677T mutation with deep venous thrombosis in Shandong Hans.

RESULTSIn case- controls, the frequencies of C/T heterozygote were 41.27% and 43.75%; whereas those of T/T homozygote were 52.38% and 36.25%. Significantly elevated mutation was observed in patients(Chi-square=6.372, P 0.01 OR(T/T)=4.552 95% confidence interval:1.440-14.390, Chi-square =6.742 P=0.009).

CONCLUSIONThe C677T mutation of methylenetetrahydrofolate reductase gene is a risk factor associated with deep vein thrombophilia in Shandong Hans.

China ; DNA ; genetics ; Gene Frequency ; Genotype ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2) ; Odds Ratio ; Oxidoreductases Acting on CH-NH Group Donors ; genetics ; Point Mutation ; Polymorphism, Restriction Fragment Length ; Thrombophilia ; enzymology ; genetics ; Venous Thrombosis ; enzymology ; genetics

OBJECTIVETo map the gene responsible for nonsyndromic hearing impairment in a consanguineous family.

METHODSFirstly, X chromosome scanning was used to exclude X chromosome. Secondly, candidate gene analyzing and genome scanning were performed by homozygosity mapping. Then, additional markers flanking the tightly linked marker were tested to confirm linkage and decide the candidate region.

RESULTSThe nonsyndromic hearing impairment of this family was autosomal recessive. Twenty-five known genes were excluded. Autosomal genome scanning indicated that D17S1293 was tightly linked with disease gene. And further study mapped the disease gene to a 5.07 cM interval bounded by D17S1850 and D17S1818.

CONCLUSIONThe disease gene of the family is mapped to a 5.07 cM interval between D17S1850 and D17S1818, which is a new locus of autosomal recessive nonsyndromic hearing impairment.

Chromosome Mapping ; methods ; Chromosomes, Human, Pair 17 ; genetics ; Chromosomes, Human, Pair 18 ; genetics ; Chromosomes, Human, X ; genetics ; Consanguinity ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Hearing Loss, Sensorineural ; genetics ; Humans ; Male ; Microsatellite Repeats ; Pedigree

OBJECTIVE: To determine the types and frequency of deafness-related variants among 7875 newborns from Dongying area of Shandong Province. METHODS: One hundred loci of 18 common deafness genes were subjected to semiconductor sequencing. Variant site, frequency and distribution of the variants were analyzed. RESULTS: In total 552 deafness gene variants were detected among the 7875 newborns, which yielded a detection rate of 7.01%. Among these, common variant sites for GJB2, SLC26A4 and GJB3 genes were c.235delC, IVS7-2A>G and c.538C>T, respectively. The variant frequencies of matrilinear inheritance deafness genes MT-CO1, MT-RNR1, MT-TL1 and MT-TS1 were 0.38%, 0.25%, 0.1% and 0.01%, respectively. Four newborns were diagnosed with deafness, among which one had unilateral hearing loss. Analysis of the proportions of neonatal deafness-related variants in five counties of Dongying showed that the highest variant rate for the SLC26A4 gene compared with GJB2 was in Lijin county (51.76% vs. 40%), while the lowest was in Hekou county (30.77% vs. 56.41%). CONCLUSION The carrier rate of deafness-related variants in Dongying area is higher than other regions of China, which may be attributed to the increased types and variant sites covered by the semiconductor sequencing method compared with the chip method and time-of-flight mass spectrometry. Due to geographical and population aggregation factors, the proportion of deafness variants in the five counties of Dongying differed significantly. Above results may provide a guide for the prevention of congenital deafness in Dongying area.