Single Nucleotide Polymorphisms in the 5'-Upstream Sequence of Chemokine Like Factor and their Association with Asthma / 中国呼吸与危重监护杂志

Objective To detect the single nucleotide polymorphisms (SNPs) in the upstream promoter region of chemokine like factor (CKLF) gene and analyze their possible associations with asthma and asthma-related phenotypes.Methods Direct Sequence of the 1553bp upstream promoter region of CKLF gene was performed in 245 Chinese Han human genomic DNAs (119 asthmatics and 126 controls).The frequencies of alleles, genotypes, and haplotypes were determined and the association of these SNPs with asthmawere further analyzed.Results Fournovel SNPs,SNP88 (T>C),SNPI96 (T>C),SNP568 (C> G) ,and SNP1047 (C > G) were found in the promoter region of CKLF.The frequency of rare allele was 0.168 (SNP88C), 0.168 (SNP196C), 0.352 (SNP568G) and 0.167 (SNP1047G), respectively.Haplotypes,their frequencies and the linkage disequilibrium coefficients between SNPs were constructed.Complete linkage disequilibrium (LDs) were observed between SNP88 and SNP196,SNP88 and SNP1047, as well as SNPI96 and SNP1047 ,respectively (D1 = 1.000,r2 = 1.000).SNP568 was in partial LD with the other three SNPs (r2 = 0.366).No association between asthma and the SNPs was observed.Conclusions Four SNPs in the regulatory region of CKLF in Chinese Han population were firstly identified.Although no significant correlation with asthma was revealed, the SNP and haplotype information is useful for other disease association studies in the future.

Programmed Cell Death 10,Beyond an Apoptosis-related Molecule / 生物化学与生物物理进展

Homo sapiens PDCD10(programmed cell death 10,alias,"TF-1 cell apoptosis related gene 15,TFAR15"),cloned by means of cDNA-representational differences analysis,had been initially identified associated with cell apoptosis.Recent research suggested mutations within the PDCD10 gene or deletion were responsible for cerebral cavernous malformations,and PDCD10 was the third CCM gene.On the other hand,other research demonstrated that PDCD10 was strictly modulated and up regulated in many kinds of tumors,which implicated that PDCD10 participated in tumorous signal transduction.The recent research confirmed that PDCD10 interacts with MST4,a member of Ste20-related kinases,and the interaction promoted cell proliferation and transformation via modulation of the ERK-MARK pathway.In conclusion,all these demonstrate that PDCD10 has many biological effects,which suggests that it is a novel player in vascular morphogenesis and/or remodeling,as well as tumorigenesis and cancer progression.

Preparation and identification of monoclonal antibodies against human programmed cell death 10(PDCD10) / 北京大学学报(医学版)

Objective: To obtain monoclonal antibodies against programmed cell death 10(PDCD10) for further study of the structure and function of PDCD10 protein.Methods: Balb/c mice were immunized with recombinant PDCD10,hybridoma cell lines secreting monoclonal antibodies against PDCD10 were screened by regular cell fusion and subcloning approach.The specificities of these monoclonal antibodies were determined by ELISA,Western blotting and Immunofluorescecence assay.Results: Three hybridoma cell lines(5G1,4F7 and 3H5) stable in secreting specific monoclonal antibodies were successfully obtained.Subclass of IgG belonged to IgG1(4F7 and 5G1)and IgG2b(3H5),respectively.The ascite titers of these monoclonal antibodies reached 1∶10~7.They could specifically bind to recombinant PDCD10 and endogenous and overexpressed PDCD10 proteins proved by ELISA and Western blotting.They failed to react with E.coli lysates and glutathione S-transferase(GST).In addition,these three monoclonal antibodies could recognize different epitopes of PDCD10 proteins assessed by immune fluorescence competitive binding assay.Both endogenous and overexpressed PDCD10 protein mainly located in the nucleus.Conclusion: Monoclonal antibodies against PDCD10 with high titers and specificity have been successfully prepared,which has laid the foundation for further study of PDCD10 protein.

Mutation detection in ED1 gene in hypohidrotic ectodermal dysplasia (HED) families / 北京大学学报(医学版)

Objective: To detect ED1 gene mutation in three hypohidrotic ectodermal dysplasia (HED) nuclear families. Methods: Peripheral blood samples were obtained from three different families of hypohidrotic ectodermal dysplasia. Genomic DNA was extracted. Polymerase chain reaction, direct sequencing and restriction enzyme reaction were performed to identify the mutations. Results: Different missense mutation in ED1 gene were found in each family: C412G, A1201G and C1375T. Two of the mutations had not been previously reported. Conclusion: Mutations in the ED1 gene are responsible for the phenotypes of HED of the patients in the family.