Functional analysis of transgenic cry1Ah-1 maize.

Maize is an important food crop in the world, but the yield and quality of maize have been significantly reduced due to the impact of insect pests. In order to address this issue, the cry1Ah gene was subjected to error-prone PCR for mutagenesis, and subsequently, the mutant cry1Ah-1 gene was introduced into maize inbred line GSH9901 callus using the Agrobacterium-mediated method. The T2 generation transformed plants were obtained by subculture, and 9 transgenic positive plants were obtained by molecular detection which was carried out by PCR, qRT-PCR, Bt gold-labeled immunoassay test strips, Western blot and ELISA. It was found that the Cry1Ah-1 gene could be transcribed normally in maize leaves, of which OE1 and OE3 had higher relative expression levels and could successfully express proteins of 71.94 KD size. They were expressed in different tissues at the 6-leaf stage, heading stage and grain-filling stage, and could ensure the protection of maize from corn borer throughout the growth period. The biological activities of OE1 and OE3 were tested indoors and in the field, and the results showed that in indoors, the corn borer that fed on OE1 and OE3 corn leaves had a mortality rate of 100 % after 3 days; in the field, OE1 and OE3 had strong insecticidal activity against corn borer, reaching a high resistance level. In conclusion, the transgenic cry1Ah-1 maize has a strong insecticidal effect on corn borer, and has a good prospect of commercialization.

[GAO Wei-bin's clinical experience in treatment of medulla oblongata paralysis with nape acupuncture].

This paper introduces GAO Wei-bin's academic thought in treatment of medulla oblongata paralysis with acupuncture. Through analyzing the etiologies and locations of medulla oblongata paralysis, in accordance with "selecting the nearby acupoints of the affected area", the acupoints are selected from the nape region, the nape acupuncture therapy and the corresponding new points are developed. Based on the human anatomy of the nape region, the anatomic structures of new points (e.g. Gongxue, Tunyan-1, Tunyan-2, Fayin, Zhiqiang and Tiyan) and their effect mechanism are explained. The treatment principle, "distinguishing the symptoms from the root causes, mutual treatment for both symptoms and root causes", is proposed, and the importance of electric stimulation of nape acupuncture is suggested in treatment of medulla oblongata paralysis.

[Reducing the maize amylopectin content through RNA interference manipulation].

To regulate the biosynthesis of maize starch, gene segment of starch branch enzyme (SBE) in maize was cloned and an expression vector of small interference RNA with the fragment of antisense gene + sense gene (pCJSBE2b, Fig.1) was constructed. The expression vector of pCJSBE2b was transformed into maize inbred lines by using pollen-tube pathway. Results of PCR amplification and Southern blotting indicated that the aimed gene with the fragment of antisense gene + sense gene had integrated into receptor maize genome (Fig.4,5) and inherited steadily in the transgenic plants (Table 1). Northern hybridization analysis showed that the aimed gene transcribed normally in the transgenic plants, which resulted in a reduction of endogenous SBE mRNA (Fig.6). The analysis of the activity of starch branching enzyme showed that the enzyme activity decreased obviously in the transgenic plants and maximum decrease was 85% compared to the control plants (Fig.7a). Although the total starch content of transgenic plants showed no change compared to the control plants but the amylose content increased to 50% (Fig.7b) in transgenic plants about 27% higher than receptor maize plants.