ObjectiveTo investigate the molecular pathogenesis of a pedigree of X-linked spondyloepiphyseal dysplasia atarda (SEDL) and to establish methods of gene diagnosis. Methods Clinical diagnosis was made based on height measurement, radiological examination and pedigree analysis. Peripheral blood samples of relevant family members were collected. After genomic DNA extraction, single strand conformation polymorphism (SSCP) followed with DNA sequencing was used to detect SEDL gene exons 36. Microsatellite marker DXS16 was selected for linkage analysis. Results The abnormal electrophoretic bands were detected in exon 4 of probands by PCR-SSCP. A c. 218C ＞ T mutation in exon 4 of SEDL gene was found in three probands, which resulted in a change in amino acid sequence S37L. The heterozygous exon 4 mutation was identified in three carriers, but not in healthy individuals, and no mutations were detect in exon 3, 5 and 6 of probands. Three unmarried young females (Ⅲ10, Ⅳ6 and Ⅳ7) were found to harbor the mutation by DNA sequencing analysis. ConclusionsA c. 218C ＞ T missense mutation in exon 4 of SEDL gene is the cause of molecular pathogenesis of the pedigree. SSCP and DNA sequencing can be used for prenatal gene diagnosis.

ObjectiveTo study the chemical constituents from the roots of Flemingia philippinensis.MethodsThe chemical constituents were isolated and purified by combination of silica gel column,Sephadex LH-20,polyamide,and ODS column chromatography.The structures of the isolated compounds were identified by means of spectral data.ResultsTen compounds were isolated from F.philippinensis and identified as isoderrone (1),dalparvin A (2),prunetin (3),7,3'-dihydroxy-5,4',5'-trimethoxyisoflavone (4),pratensein-7-O-β-D-glucoside (5),sissotrin (6),sophororicoside (7),formononetin (8),orobol (9),and biochanin A (10).ConclusionCompounds 1-6 are obtained from this plant for the first time.

OBJECTIVETo identify the gene causing hereditary multiple exostoses in a Chinese pedigree.

METHODSLinkage analysis was carried out in the family using microsatellite markers close linkage to the EXT1 and EXT2 genes to define the candidate gene. Then the whole coding sequence and the intron-exon boundaries of the candidate gene were amplified and sequenced.

RESULTSThe disease-causing gene of the family was linked to the EXT2 gene. A nonsense mutation of 536G>A in exon3 of the EXT2 gene was detected, which was co-segregated with the disease phenotype. The mutation resulted in a stop codon in codon 180. A nonpenetrant case was found in the family.

CONCLUSIONThe mutation 536G>A in the EXT2 gene is the disease-causing mutation in the pedigree with hereditary multiple exostoses.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Child, Preschool ; China ; Codon, Nonsense ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; N-Acetylglucosaminyltransferases ; genetics ; Pedigree ; Young Adult

Objective To screen the hemostatic active extracts from Callicarpa nudif lora .Methods The powdered Callicarpanudiflora was extracted with 70% EtOH and concentrated to give EtOH-extract .The EtOH-extract was further chromatographed over HP-20 macroporous resin column ,eluting with aqua and 95% EtOH to get HP-H2 O-elution and HP-EtOH-elution ,respectively .The obtained three extracts of EtOH-extrract ,HP-EtOH-elution and HP-H2 O-elution were set as large ,middle and small dosage groups for drug preparation ,respectively .Yunnan Baiyao was used as a positive control group . The weight increment ,bleeding time and clotting time of fed mice were detected by cutting tail and grass slide methods after in-tragastric administration for 7 days .Results As compared with blank model and positive control ,each dosage groups of HP-EtOH-elution could significantly shortened the bleeding time ,of which the small dosage group and middle dosage group even ex-hibited better results than the positive control group .Whereas the EtOH-extract ,HP-EtOH-elution and HP-H2 O-elution didn′t demonstrated significant effect on clotting time as well as the weight increment .Conclusion The HP-EtOH-elution was suggested to be the major active extract possessing hemostatic activity to mice under tested dosages .The hemostatic mechanism may through in-trinsic coagulation pathway .This study would be helpful for further phytochemical investigation of C .nudi f lora .

OBJECTIVETo explore the possible roles of polymorphisms of SPO11 and glutathionine S-transferase (GST) genes in idiopathic male infertility in a ethnic Han Chinese population from Henan.

METHODSMultiplex PCR and DNA sequencing were performed to determine the SPO11 c.517C>T(rs28368082) and GST genes (GSTM1, GSTT1, GSTP1) polymorphisms in 216 idiopathic male infertility cases and 198 normal samples.

RESULTSThe frequencies of the SPO11 CC and CT genotypes were 87.5% (189/216) and 12.5% (27/216) in the patients, and 97.5% (193/198) and 2.5% (5/198) in the controls, respectively. The frequencies of SPO11 CC and CT genotypes, the A>G transition at nucleotide 313 in the exon 5 of the GSTP1 gene, and the frequencies of combined genotypes GSTM1 (-/-), GSTT1 (+/+), GSTP1 (AA) and SPO11 (CT) were significantly different between the two groups (P<0.05).

CONCLUSIONThe rs28368082 polymorphism of the SPO11 gene, the A>G transition at nucleotide 313 in the exon 5 of the GSTP1 gene, and the combined genotypes of GSTM1 (-/-), GSTT1 (+/+), GSTP1 (AA) and SPO11 (CT) may be associated with idiopathic male infertility in ethnic Han Chinese.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Endodeoxyribonucleases ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; ethnology ; genetics ; Genotype ; Glutathione S-Transferase pi ; genetics ; Glutathione Transferase ; genetics ; Humans ; Infertility, Male ; enzymology ; ethnology ; genetics ; Linkage Disequilibrium ; Male ; Mutation ; Odds Ratio ; Polymorphism, Genetic ; Sequence Analysis, DNA

Objective To investigate the chemical composition from the flowers of Callerya speciosa,and reveal the metabolites difference at different flowering periods based on metabolomics technology.Methods The primary and secondary metabolites,volatile chemical components in flowers of C.speciosa were analyzed combined by GC-MS and UPLC-Q-Exactive MS.Principal component analysis(PCA),orthogonal partial least squares-discriminant analysis(OPLS-DA),and hierarchical cluster analysis(HCA)were performed to identify differential metabolites.Results A total of 332 compounds were identified by UPLC-Q-Exactive MS,mainly including secondary metabolites such as flavonoids,triterpenoids,phenylpropanoids.A total of 297 compounds were identified by GC-MS,mainly including primary metabolites and volatile chemical components,such as organic acids,amino acids,saccharides,heterocycles,alcohols.The PCA analysis demonstrated that the metabolites of the four flowering periods were divided into two groups:bud,initial bloom and blooming periods clustered into one group,while wilting period clustered into the other group,the main differences were filtered and identified as flavonoids and triterpenoids,organic acids,respectively.Compared to the upright type,the flowers of vine type contained more characteristic flavonoids as differential metabolites during the bud,initial bloom and blooming periods,and some flavonoids decrease gradually with the development of flowering.Conclusion The results indicated that the flowers of C.speciosa possessed abundant active flavonoid metabolites for further utilization,and the best harvest stage is initial bloom,the best harvest plant is vine type.This study provides a scientific basis for the scientific development and rational use of the flowers of C.speciosa.