Effect of Down-Regulating the CD59 by RNAi Lentivirus on the Expression of Acute T-lineage Leukemia Jurkat Cell Line / 中国实验血液学杂志

OBJECTIVE@#To analyze the effect of down-regulating the CD59 gene expression by RNAi lentivirus as vector on Jurkat cell line of acute T-lineage leukemia.@*METHODS@#The expression of CD59 in Jurkat cell line of acute T-line leukemia was induced to decrease by RNAi lentivirus as vector. The transfection of RNA lentivirus and the localization of CD59 molecule were analyzed by laser confocal technique. The relative expression of CD59 gene in blank control, negative control and RNAi lentivirus transfected group was detected by real-time fluorescence quantitative PCR, and the enzyme-linked immunosorbent assay was used to detect the expression of TNF-β and IL-3 in supernatants of cultured cells in 3 groups. The expression levels of apoptosis-related molecules including Caspase-3, Survivin, BCL-2 and BCL-2-associated X protein (BAX) were measured by Western blot.@*RESULTS@#The transfection efficiency for Jurkat cells was higher than 90%. CD59 was mainly located on the cell membrane. Compared with the blank control group and the negative control group, the expression level of CD59 mRNA and protein in the RNAi lentivirus transfected group significantly decreased (P<0.05). Compared with the blank control group and the negative control group, the expression of TNF-β and IL-3 in the RNAi lentivirus transfected group were significantly higher and lower (P<0.05) respectively. The expression levels of Survivin and BCL-2 in the RNAi lentivirus transfected group were significantly lower than those in the blank control group and the negative control group, while the expression levels of Caspase-3 and BAX in the RNAi lentivirus transfected group were significantly higher than those in the blank control group and the negative control group (P< 0.05).@*CONCLUSION@#The down-regulation of CD59 gene expression induced by RNAi lenti-virus can decrease the expression of proliferation and differentiation-promoting molecule such as IL-3 and increase the expression of TNF-related factor in Jurkat cell line of acute T-lineage leukemia, which also can increase the expression of apoptosis-related proteins such as Caspase-3 and BAX, and decrease the expression of anti-apoptosis-related proteins such as Survivin and BCL-2.

[Mapping and cloning of GAD1-2 for long awn using CSSLs in rice (Oryza sativa L.)].

Rice is an important food crop in the world. The awn may protect rice seeds from being cut by birds, which is important in rice domestication, survival and diffusion. However, the characteristic of awn is gradually washed out during rice domestication and artificial selection. Mapping and cloning of rice awn genes is the basis of studying the genetic mechanism of awn domestication. In this study, 146 chromosome segment substitution lines (CSSLs) derived from DongNanHui 810/ZhangPu wild rice with DongNanHui 810 as the recurrent parent were used to analyze the quantitative trait loci (QTL) controlling the long awn of rice. The results showed that four CSSLs contained one QTL for the long awn. Using substitution mapping, the GAD1-2 gene was mapped between two markers (Ind8-10 and RM4936) on chromosome 8, with a genetic distance of about 4.75 Mb. Using the dominant individuals of segregating populations, the GAD1-2 gene was eventually located between two Indel markers, with a physical distance of about 27 kb, which contained only two candidate genes Os08g0485500 and Os08g0485400. Sequencing analysis showed that Os08g0485500 was the candidate gene of GAD1-2. Further analysis showed that there were six bases missing in the conservative ORF region, resulting in the absence of serine and cysteine that led to the long awn of the four CSSLs. The GAD1 gene was also cloned in this position, suggesting that GAD1-2 and GAD1 were allelic. This study laid a foundation for further understanding of the genetic regulation mechanism and genetic evolution of the awn gene in rice.

[Morphological characteristics and gene mapping of a palea degradation(pd2) mutant in rice].

The yield and quality of rice are directly impacted by floral organ development in rice. Understanding of the floral development mechanism will be useful in genetic improvement of yield and quality. In this study, a rice mutant palea degradation 2 (pd2) in an indica cultivar '8PW33' was obtained after 60Co γ-ray treatment. Analysis of the mutant showed that, compared to the wild type, plant height, total grain number per panicle, and sword leaf width were significantly increased, but the seed setting rate were significantly decreased. The florets of the mutant exhibited degraded palea and sickle-shaped tortuous lemma. Detail examination using scanning electron microscopy revealed that when epidermis of the vane and lemma were normal, epidermis of the palea were arranged tightly, which might result from degraded palea. Genetic analysis supported that this mutation phenotype was controlled by a single recessive gene. Polymorphic analysis of simple sequence repeat markers demonstrated that PD2 gene is located on chromosome 9. With a larger mapping population and more indel markers, we further mapped PD2 gene between 2 indel markers with a physical region of about 82 kb. Within this region, there is a cloned gene REP1 known to control rice palea development. By comparing the DNA sequences of REP1 from pd2 and 8PW33, in combination with the results of phenotypic analysis, we concluded that PD2 is an allele of REP1.

[Study of the pH sensitive polymer].

A pH sensitive polymer was prepared by copolymerization of methacrylic acid as monomer, diethylene glycol dimethacrylate as cross-linking reagent, heptane as porogen, and fluorescent dye eosin as indicator. The factors of influence on the preparation, and the character of the pH sensitive polymer for pH were studied. The maximal emission wavelength of eosin was red shifted in the polymer than in solution, the apparent Ka largened, and the dissociation equilibrium of indicator was shifted to acidity direction, because the polarity of polymer diminished. Under the optimal condition, the calibration curve of the pH sensitive polymer covered the range of pH 0-3.0 with good reproduction and reversibility.

Study on mechanism of the anti-tumor activity of Acanthopanax gracilistylus / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the mechanism of anti-tumor activity of Acanthopanax gracilistylus extract (Age).</p><p><b>METHODS</b>The tumor cells proliferation was detected by using (3H)-TdR incorporation method, and the effects of Age on cell cycle of tumor cells, retinoblastoma (Rb) protein and cyclin-dependent kinases (Cdk) were analyzed by flow cytometry and Western blotting assay, respectively.</p><p><b>RESULTS</b>It was indicated by cytoactivity test in vitro that Age only had effect in inhibiting the proliferation of tumor cells, it couldn't lead to death of cells. Under action of Age, the proliferation of tumor cells was halted at G0/G1 stage of cell cycle, and showed no direct cytotoxic effect by Age. Age could induce lowering of the expression of Rb, Cdk2 and Cdk4, cause halt of tumor cell proliferation.</p><p><b>CONCLUSION</b>The tumor inhibitory effect of Age is realized by way of regulating the activity of cell cycle controlling enzymes to suspend the proliferation of tumor cells.</p>