Effect of interferon on HEL cell apoptosis and JAK2 V617F mutation gene expression / 实用医学杂志

Objective To observe the effect of Interferon-α2b on HEL cells (human erythroleukemia cell line) growth, apoptosis and JAK2 V617F mutation gene expression. Methods HEL cells were placed in RPMI1640 containing 10% FBS and incubated in a cell incubator. Cells in the logarithmic growth phasem were collected, adjusting the cell density to 1 × 105/mL for experimental research. The interferon concentration in five groups were 0, 5 × 105, 10 × 105, 50 × 105, 100 × 105 U/L, with different incubation time (0, 24, 72, 120 h), respectively. The cell growth status in different groups was observed in the inverted optical microscope; MTT was used to detect the inhibition of interferon on HEL cell proliferation. Cell apoptosis was detected by flow cytometry. Fluorescence quantitative PCR was used to detect the mutation gene of JAK2 V617F expression. Results Inhibition rates of Interferon on the HEL cell proliferation in 5 × 105 U/L, 10 × 105 U/L, 50 × 105 U/L, 100 × 105 U/L groups were 18.57%, 25.10%, 42.10%, 57.00%, respectively. JAK2 V617F/GAPDH by fluorescence quantitative was 1.556, 1.213, 0.870 respectively under the concentration of interferon 100 × 105 U/L for 24, 72, 120 h. Conclusions Interferon-α2b can inhibit HEL cells proliferation and induce HEL cells apoptosis. Increasing concentration of interferon increases HEL cell apoptosis rate. Interferon can inhibit JAK2 V617F expression of HEL cells in a dose-dependent manner.

Advances in gene chips for early gastric cancer / 国际外科学杂志

Gastric cancer incidence is one of the most common malignancies in our country and is the second most common in the worldwide,clinic doctors always emphasize early diagnosis and treatment of gastric cancer patients,in order to reduce the mortality,however,most patients' condition often have been in the late fall and these patients were badly in efficacy.Looking for a new diagnosis way is a medical prddem,with molecular biology advance and gene chips was improved,it is possible for the early screening of gastric cancer.This assay aims to briefly analyse the role of gene chips in the research progress of early gastric cancer.

Expression of Cystic Fibrosis Transmembrane Regulator in Nasal Polyps / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Nasal polyps are also prominent features of cystic fibrosis. The purpose of this study is to investigate the CFTR expression and CF mutation genes in nasal polyps to verify genetic influence in the nasal polyp formation. MATERIALS AND METHODS: We have evaluated 30 nasal polyps, 10 recurrent nasal polyps, and 10 inferior turbinates. RT-PCR was done for the CFTR mRNA expression and mutaion genes were studied by RFLP. Immunohistochemical study and western blotting were done for CFTR expression. RESULTS: RT-PCR revealed no differences in the expressions of CFTR transcripts between nasal polyps and nasal turbinates. The expression of CFTR protein was localized on apical portion of some ciliated cells on immunohistochemistry, and western blotting showed no differences in expression levels of CFTR protein. Three different mutations (deltaF508, 591 del 18, G551D) were analysed. One case of deltaF508 was detected in the samples. CONCLUSIONS: The expression levels of CFTR mRNA and CFTR protein may not be associated with the pathogenesis of nasal polyps, but it needs to be studied further on the physiological base. We also need to study further regarding the relation between CFTR mutaion genes and the development of nasal polyps with more mutaional screenings in cDNA levels.

Cloning of human ? thalassemic mutation ? IVS II654(C→T) and establishment of its mammalian expression system / 中国病理生理杂志

AIM: To clone human ?-globin gene carrying a thalassemic mutation IVS II654(C→T) and establish a eukaryotic expression system for high-level expression of human ? IVS II654 gene in mouse erythroleukaemia(MEL) cells. METHODS: The fragments of human ? 654 gene isolated from the ? thalassemia patients homozygous for the ? 654 mutation were amplified by PCR, and cloned to plasmid pBGT51. Then, the human ? LCR and ? 654 gene were subcloned from plasmid pBGT51 to the stable mammalian expression vector pcDNA3.1+ together, and the MEL cells were transfected with this vector using commercially available cationic lipid FuGENE6. The MEL cells were induced for further maturation by DMSO and the expression of human ? 654 gene in the MEL cells was identified by RT-PCR. RESULTS: A mammalian expression system of human ? thalassemic mutation ?IVS II654(C→T) was established. CONCLUSION: The level and the reliability of expression of human ? 654 gene in the MEL cells in vitro are similar to that in vivo in human body. This may be a valuable gene therapy model for human ? thalassemic mutation ?IVS II654(C→T).