ABSTRACT
Objective To observe apoptosis of primary cultured synoviocytes from patients with rheumtoid arthritis (RA) which induced by transfected recombinant plasmid carring Fas ligand (FasL) gene, aimed to detect the aim targets and develope gene therapy for RA by intra-articular injection. Methods FasL cDNA was introducted into the plasmid pcDNA3.1-neo by reverse transcriptase-polymerse chain reaction (RT-PCR) to set up the recombinated human FasL gene expression carrier, Then the pcDNA3.1-FasL was transducted into the primary cultured synovial cells by catholyte liposomes. After screened by G418, transgenic synovial cells were choosen. Finally through the microscope, TUNEL, Annexin Ⅴ-FITC and PI staining with flow cytometry, we observed the apoptosis state of each synovial cell. Results Prokaryotic and eukaryon carrier with FasL gene were established successfilly. The gene sequencing had completely coincident with the lierature. When eukaryon carrier with FasL (pcDNA3.1-FasL) was transducted into the index number livinging RA synovial cells after 15 h, most synovial cells presented distortion transformation, and the minority sheded from the bottom wall. The transgenic RA synovial cells after screened by G418, only a little amount survived about 1~2 per sight, and the cells grew slowly, the normal matched cells were cloned after 2~4 weeks. The FasL gene transgenic RA synovial cells presented positive findings by TUNEL staining. Early apoptosis cells by Annexin Ⅴ/PI staining, but the difference wasn′t significant. Conclusion The eukaryon carrier with FasL (pcDNA3.1-FasL) can be transferred into RA synovial cells. It can induce synovial cell apoptosis.
ABSTRACT
Objective To study the relationship between KAI1 expression and metastasis of colon can cer. Methods KAI1 mRNA expression was determined by Northern blot anal ysis and in situ hybridization in 10 normal colon and 24 colorectal cancer tissu es. Results By Northorn blot analysis, there was a wide range of KAI1 mRNA expression levels in both normal and cancerous tissue samples. Primary colo n cancers with lymph node metastasis exhibited significantly lower KAI1 mRNA e xpressio n levels than primary tumors without lymph node metastasis(P
ABSTRACT
By using single spleen colony transplantation technique and sex chromosome typing as natural cytogenetic markers, we have been able to demonstrate that most spleen colony forming cells in the adult bone marrow or in foetal liver of inbred LACA or C57 mice possess the potential of re-establishment of myeloid and lymphoid lines of cells in lethally irradiated mice. However, most of the spleen colony forming cells in the peripheral [blood of normal mice possess weak or no potentiality to proliferate or re-establish hemopoiesis in lethally irradiated mice. Our results support the notion that the CFU-S population in mice is heterogeneous.From the nature of colony forming cells in the peripheral blood of normal mice, we are convinced that the assessment of CFU-S content relying upon the spleen colonies as the unique sign to indicate the self-renewal ability of a spleen colony forming cell is by no means valid inasmuch as some plu-ripotent . hemopoietic progenitors derived from pluripotent hemopoietic stem cells may have the same ability to form colonies in vivo.