AIM:To construct a recombinant eukaryotic expression vector pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/? and detect its expression in NIH 3T3 cells.METHODS:CD28-? cDNA was amplified from the plasmids pBULLET and inserted into pLNCX vector that contained anti-CD20 scFv/IgGFc/CD80 gene.The recombinant plasmids were transfected into NIH 3T3 cells,and resistant clones were obtained by G418 selection.The gene expression of the fusion protein was determined by RT-PCR and FACS.RESULTS:The recombinant eukaryotic vector was constructed successfully,determined by PCR and enzyme digestion analysis.The target gene was amplified from NIH 3T3 cells transfected with the vectors by RT-PCR.The FACS showed that recombinant protein was expressed in NIH 3T3 cells.CONCLUSION:Construction of pLNCX/anti-CD20scFv/IgGFc/CD80/CD28/? expression vector and its expression in NIH 3T3 cells lay the foundation for further research of generation of modified T lymphocytes to CD20 positive lymphoma.

The required time of conventional magnetic resonance spectroscopic imaging technique is too long to be applied to clinic. It is necessary to develop the fast methods for magnetic resonance spectroscopic imaging. Nowadays there are 7 kinds of methods presented, which come from MRI techniques. In this contribution the conventional spectroscopic imaging and 7 sorts of fast spectroscopic imaging are elaborated. It is envisaged that more rapid imaging techniques will be designed, if these arbitrary trajectory reconstruction methods in MRI are applied to spectroscopic imaging.
Magnetic Resonance Imaging ; methods ; trends ; Magnetic Resonance Spectroscopy ; methods

A PC-based software was developed and programmed with VC++6 for reconstructing MR images from the data acquired on an irregular k-space trajectory. It can read clinical MRI raw data and image data, create numerical phantoms, design k-space trajectories, generate k-space data from numerical phantom, calculate weighting functions, reconstruct images, and carry out error analysis for the reconstructed images. It is helpful to the investigations of new k-space trajectories and new reconstruction algorithms.
Algorithms ; Bayes Theorem ; Brain ; anatomy & histology ; Fourier Analysis ; Humans ; Image Processing, Computer-Assisted ; methods ; Magnetic Resonance Imaging ; methods ; Programming Languages ; Software ; Software Design

OBJECTIVETo screen for KIT gene mutations in two Han Chinese pedigrees affected with Piebaldism.

METHODSClinical data of the pedigrees was collected. Genomic DNA was extracted from blood samples collected from the pedigrees and 120 unrelated healthy controls. All coding exons of the KIT gene were subjected to PCR amplification and direct sequencing.

RESULTSTwo missense mutations, c.1861G>A(p.Ala621Thr) and c.1872G>A(p.Met624Ile), were identified respectively in the two pedigrees. Neither mutation was found among healthy members from the respective pedigree and the 120 unrelated healthy controls. c.1872G>A is a novel mutation.

CONCLUSIONMutations of the KIT gene may affect the structure and function of the transmembrane receptor KIT, which lead to the disease.