BACKGROUND: 1200009K10 gene is from the lung cDNA pool of adult mouse and announced in January 2002, but its function is still not known.The predicted protein has five domains: Kelch motif, Kelch domain, BTB/POZ domain, BTB domain and β propeller domain, all of which are involved in protein-protein interactions and some enzymatic activities.OBJECTIVE: To explore the correlation between the expression of 1200009K10 gene and retinitis pigmentosa in mice.DESIGN: Observational and comparative trial.SETTING: Institute of Zhongshan Ophthalmic Center of Sun Yat-sen University.MATERIALS: The rd mice, rds mice and passage mice from C3B mice as normal control were purchased from Jackson Laboratory (Bar Harbor,Maine 04609, USA), and raised in second class unit of Experimental Animal Center of Sun Yat-sen University.METHODS: The retinal RNA of mice was extracted and isolated and 1200009K10 gene was amplified by rapid amplification of eDNA ends method (RACE). The gene expressions in the retina of rds, rd and C3B mice were analyzed, respectively by gene-specific real-time quantitative PCR.MAIN OUTCOME MEASURES: The expression contents and patterns of 1200009K10 gene in retina of three kinds of mice.RESULTS: One clone was obtained after RACE of retinal RNA extracted from retina of mice, which had an almost identical sequence with the 1200009K10 gene. Expression of 1200009K10 gene was classified into 6stages: 7, 12, 25, 37, 50 (near sex maturity) and 150 days after born. The expressions of 1200009K10 gene in the retina of rds, rd and C3B mice were low at postnatal day 7 (P7), higher between P7-12, stable between P12-50, and finally increased a little. The expression trend and level of 1200009K10 gene of C3B mice was nearly the same with that of the rds and rd mice, and higher than that of rds and rd mice at P12. Although there were differences in expression levels of 1200009K10 gene among the three kinds of mice, the expression patterns were almost the same.CONCLUSION: 1200009K10 gene may not participate in the development of retinal degeneration.

AIM: To identify the genes that were differentially expressed in the retina of rds mouse during the development of retinitis pigmentosa.METHODS: mRNA differential display method was used to compare and analyze mRNA samples prepared from the retina of rds mouse and normal mouse on postnatal day 25(P25). Differentially expressed fragments were cloned, sequenced and compared with GenBank database by BLASTN. Expression difference was further investigated by gene-specific primer RT-PCR.RESULTS: Obvious difference in gene expression occurred between rds mouse and normal mouse. One fragment, clone No.5, shared 91% homology with rat NADH-cytochrome b5 reductase (b5R) cDNA. Thus, it was identified as mouse b5R cDNA. Gene-specific RT-PCR confirmed that b5R mRNA level was increased in the retina of rds mouse compared with normal mouse on postnatal day 12, 25 and 37, respectively.CONCLUSION: Certain oxidative factors may up-regulate the expression of b5R resulting in large consumption of NADH and production of NAD +, through which apoptotic retinal cell death was enhanced.

Objective To investigate the effect and mechanism of cell differentiation agent Ⅱ (CDA-Ⅱ) on the differentiation of human acute mycloid leukemia (APL) HL-60 cells.Methods The cell morphology and differentiation was detected by Wright-Giemsa staining,the expression of cell surface differentiation antigen CD11b of HL-60 was detected by flow cytometry,the differentially expressed genes were screened by gene expression profile chip (NimbleGen).Results The result of Wright-Giemsa staining showed that CDA-Ⅱ induced HL-60 differentiation towards mature stages in a time-dependent manner.After treated with CDA-Ⅱ,the percentage of CD11b-positive HL-60 cells significantly increased in a time-and dose-dependent manner.The result of gene expression profiling indicated differentially expressed genes including 113 up-regulation genes and 140 down-regulation genes.The up-regulation expression genes involved in six pathways including mineral absorption,complement and coagulation cascades,down-regulation expression genes involved in nine pathways including pyrimidine metabolism,RNA polymerase,purine metabolism and so on.Conclusion CDA-Ⅱ can induce HL-60 differentiation and make gene differentially expressed.The data provide the feasibility of CDA-Ⅱ in differentiation induction therapy for APL.

AIM: To understand the effect of the RB1 gene mutation on the function of pRB (the protein product of the RB1 gene) in the patients with retinoblastoma (RB). METHODS: The genomic DNA from retinoblastoma patients was extracted. After amplification, the promoter and all 27 exons were screened by SSCP-heteroduplex method. The mutation was cloned and identified by sequencing. The effect of the mutation product on the function of pRB was analyzed. RESULTS: One missense mutations of the exon 4 of the RB1 gene was identified in the genomic DNA from RB patients. This mutation was outside the large pocket of the pRB. No mutation of the RB1 gene was found in the genome DNA of the patient's parents. This is the fourth report that there was a genome mutation located outside the large pocket of pRB in the RB patients. CONCLUSION: The amino-terminus of the pRB may be essential for growth suppression.

Objective To investigate the effects of shRNA interference (RNAi) targeting the histone lysine specific demethylase 1 (LSD1) on the proliferation and apoptosis in acute leukemia (AL) cells. Methods LSD1 shRNA vectors were constructed and transduced into HL-60 and SHI-1 AL cell lines. Inhibitory efficiency of LSD1 was detected by real-time quantitative PCR (RT-qPCR) and Western blot respectively. Cell proliferation was assessed by methyl thiazolyl tetrazolium (MTT), and cell apoptosis was measured by flow cytometry. Results After interference of LSD1, the expression levels of LSD1 mRNA and protein in HL-60 and SHI-1 cells (mRNA: 0.242 ±0.023, 0.207 ±0.006; Protein: 0.256 ±0.015, 0.486 ±0.042) were decreased compared with blank control group without transfection process (mRNA: 1.021 ±0.178, 1.039 ±0.395; Protein:0.552 ±0.026, 0.754 ±0.060) and empty vector negative control group (shNC group) (mRNA: 0.935 ±0.136, 1.016±0.203;Protein: 0.500±0.026, 0.750±0.049) (P<0.05). The levels of their cell proliferation (absorbance value: 0.712±0.010, 0.549±0.007) were inhibited compared with blank control group (absorbance value:0.823±0.010, 0.625±0.005) and shNC group(absorbance value: 0.818±0.019, 0.621±0.003) (P< 0.05). While cell apoptosis rates were increased [(32.80 ±1.35) %, (23.49 ±1.40) %] compared with blank control group [(8.08 ±0.62) %, (7.28 ±1.17) %] and shNC group [(8.08 ±0.62) %, (7.28 ±1.17) %] (P< 0.05). Conclusions Lentivirus-mediated shRNA interferencing LSD1 can inhibit cells ' proliferation and promote apoptosis of HL-60 and SHI-1 AL cell lines, indicating that LSD1 may be a potential biological molecular marker and a new treatment target for AL.