Detection and analysis of an ATP2A2 mutation in a family with Darier-White disease / 中华医学遗传学杂志

OBJECTIVE@#To explore the molecular basis for a pedigree affected with Darier-White disease.@*METHODS@#Genomic DNA was isolated from 3 patients and 1 unaffected member from the pedigree, as well as 80 healthy controls. Targeted sequence capture and next-generation sequencing were used to screen mutations of skin disease-related genes. Candidate mutations were verified by Sanger sequencing, and co-segregation analysis was carried out to confirm the pathogenicity of mutation. Conservation analysis and protein structure and function were also predicted with Bioinformatic tools.@*RESULTS@#A heterozygous mutation c.2246G>T (p.G749V) was identified in exon 15 of ATP2A2 gene in all 3 patients from the pedigree, but not in the unaffected member or 80 healthy controls. The corresponding amino acid was highly conserved, and mutation of which can lead to structural and functional changes of the protein.@*CONCLUSION@#The c.2246G>T missense mutation of the ATP2A2 gene probably underlies the Darier-White disease in this pedigree by causing damages to the structure and function of sarcoplasmic/endoplasmic reticulum calcium ATPase 2 (SERCA2).

"The teaching exploration of common optional course""forensic medicine"" for clinical medicine undergraduate students" / 中华医学教育探索杂志

In teaching forensic medicine as a common optional course for clinical medicine undergraduate students, we carried on the beneficial attempt in the overall set-up of teaching program, the effective implementation of the case-based and discussion-based teaching methods, the use of multimedia and internet classroom, plentiful of teaching models such as demonstration and practice, discussion forums between teachers and students etc. Through these efforts, we enhanced the teaching quality of the public elective course offorensic medicine.

Screening of binding proteins interact with phosphotyrosine-interacting domain of DOC-2 by yeast two hybrid system / 中国肿瘤生物治疗杂志

Objective: To screen for proteins which can interact with phosphotyrosine-interacting domain (PID) of differentially expressed gene in human ovarian cancer cell line DOC-2 by yeast-two hybrid technique, so as to provide evidence for the signal pathway of DOC-2. Methods: The cDNA sequence of human DOC-2 gene was amplified and its PID domain (nDOC-2) was subcloned into the bait vector pGBKT7 of yeast two-hybrid system; the product was then used to screen an embryo brain cDNA library and the proteins interacting with nDOC-2 were identified. Quadrople dropout(QDO) medium and X-?-gal were used for selecting the positive clones. PCA was used to analyze the amplified sequence. After elimination of the false positive clones, the positive clones were sequenced and analyzed by bioinformatic methods. Results:Twenty-one candidate positive clones were obtained and 3 of them were plasmids encoding Homo sapiens partial mRNA for betaglycan (TBR III gene), Homo sapiens protocadherin gamma subfamily C 3 (PCDHGC3), and APLP1(amyloid beta precursor-like protein 1).Conclusion: The proteins obtained in this study may play important roles in the signal pathway of DOC-2, which provides a new orientation for DOC-2 gene therapy of ovarian cancers

Preparation of rabbit-anti human ERA serum / 细胞与分子免疫学杂志

Aim To express h-ERA and h-ERA C-terminal domain in E.coli and prepare antiserum against human ERA. Methods Human era(h-era) cDNA and h-ERA C-terminal domain gene were amplified by PCR and ligated with prokaryotic expression vector pRSET-C and pDH respectively. E.coli TAP106 transformed with the recombinant plasmid pDH-h-era-C was induced at 42℃ to express h-ERA C-terminal domain protein. Antiserum against h-ERA was prepared by immunizing rabbit with h-ERA C-terminal domain protein purified by SDS-PAGE. E.coli BL21(DE3) transformed with the recombinant plasmid pRSET-C-h-era was induced with IPTG to express (His)6-h-ERA fusion protein for Western-blot analysis.Results The expressed h-ERA C-terminal domain and (His)6-h-ERA fusion protein occupied about 40％ and 80％ of total bacterial protein respectively. The(His)6-h-ERA fusion protein expressed in E.coli can be detected with the rabbit antiserum. Conclusion The h-ERA C-terminal domain protein and (His)6-h-ERA fusion protein were expressed with high efficiency in E.coli, and the rabbit antiserum against h-ERA was prepared successfully.