Gene transformation by in situ pistil delivery method in Brassica

Agrobacterium-mediated gene transformation in Brassica crops is a time-con-suming and labor-cost method （tissue culture and plant regeneration） in a genotype-depen-dent way. Here we described a new method 7n situ pistil delivery （ISPDy method with no tissue culture and adaptive to three Brassica species. Briefly this method can be stated as placement of a liquid drop containing binary vector DNA onto the cut surface of emasculated and artificial pollinated Brassica stigma. Transformation rate （Basta-resistant seedlings per 100 pistils） of different species ranged from 0.4% to 16.3%. Factors affecting transforma-tion success included tinning and buffer type. PCR and Southern blot confirmed transgenic events. Reciprocal cross showed reporter gene steadily inherited in nuclear transformed pattern. GUS histochemical assay of transgenic immature seeds showed stained embroys and endosperm in the same seed, indicating DNA delivered through style and integrated into Brassica seed/genome after double fertilization.

Efficiency of Different Methods for dsRNA Delivery in Cotton Bollworm(Helicoverpa armigera)

RNAi trigged by dsRNA not only facilitates the development of molecular biology, but also initiates a new way for pest control by silence of fatal genes. However, one of the key limitations in pest control is lack of the convenient and efficient method for dsRNA delivery. In this study, different dsRNA delivery methods at their own optimum conditions were evaluated comparatively for their efficiency with Helicoverpa armigera as test animal. It was found that the popular one- time injection of larvae with dsRNA could reduce the pupation rate by 43.0% and enhance larva mortality by 11.7%. One- time ingestion of dsRNA did not result in any significant effect on phenotype. Continuous ingestion of in vitro synthesized dsRNA by refreshing the bait diet every day caused 40.4% decrease in successful pupation and 10.0% increase in larval mortality, which was similar as one-time injection. The most efficient method was found to be the continuous ingestion of the bacteria containing dsRNA expressed, which reduced the rate of pupation by 68.7% and enhanced the larval mortality by 34.1%. Further analysis found that dsRNA was degraded faster in midgut juice than in hemolymph. However, the cell of bacteria could protect dsRNA and delay the degradation in the midgut juice of H. armigera. These results throw light on the application of dsRNA in pest management with proper ways.

The Pubic Arch Angle： A Study on the Predictability of Labor and Delivery

Study objective： to verify if the PAA （angle of the pubic arch） is a predictive data for the mode of delivery, the duration of labour and the expulsion period. Methods： we chose to measure the PAA in 100 women recruited, chosen from the 38th to the 42th week of gestation. The method involves the use of two-dimensional transperineal ultrasound. Data collection took place over a period of time from May to September 2015. The ante-partum data taken into consideration were the patient’s name and surname, PAA, parity, age, height; those acquired in the post-partum provide for the calculation in minutes of the active phase of labor, the duration of the expulsion period, the execution of the amniorrhexis and the use of the synthetic oxytocin, the fetal cranial circumference, the fetal weight and the position of the presented part. Results： results discovered ultrasoundly 100 PAA, with an average value of 115.5° and we proceeded by evaluating 90 vaginal births. Infants at birth had a CC with an average value of 34.5 mm. By relating the two variables PAA and CC for the dependent variable “period expulsion in minutes”, confirming that the expulsive period depends on the relationship between PAA and CC. The average of the minutes of the expulsion period is 50＇ in women with a PAA 〉 111.5° and a CC 〉 34.5 mm while in women with an PAA 〉 111.5° the media time is 30＇ regardless of the CC. It has been shown that oxytocin and amniorrhexis reduce the minutes of the expulsion period only in a case of PAA 〉 111.5° and a CC 〉 34.5 mm bringing them from 60＇ to 23＇. Conclusion： the duration of the active phase of labor is not influenced by the PAA but it depends on external factors. In the case of the expulsion period, a predictability of the PAA on its duration is shown if the CC is taken into consideration; as regards the influence of the PAA on the method of delivery, it was not possible to carry out any analysis since all the useful cases had vaginal delivery.

Novel nanoparticles composed of chitosan and β-cyclodextrin derivatives as potential insoluble drug carrier

This research was aim to develop novel cyclodextrin/chitosan （CD/CS） nanocarriers for insoluble drug delivery through the mild ionic gelation method previously developed by our lab. A series of different β- cyclodextrin （β-CD） derivatives were incorporated into CS nanoparticles including hydroxypropyl-β- cyclodextrin （HP-β-CD）, sulphobutylether-β-cyclodextrin （SB-β-CD）, and 2,6-di-O-methy-βcyclodex- trin （DM-β-CD）. Various process parameters for nanoparticle preparation and their effects on physicochemical properties of CD/CS nanoparticles were investigated, such as the type of CD derivatives, CD and CS concentrations, the mass ratio of CS to TPP （CS/TRP）, and pH values. In the optimal condition, CDICS nanoparticles were obtained in the size range of 215-276 nm and with the zeta potential from 30.22 mV to 35.79 mY. Moreover, the stability study showed that the incorporation of CD rendered the CD/CS nanocarriers more stable than CS nanoparticles in PBS buffer at pH 6.8. For their easy preparation and adjustable parameters in nanoparticle formation as well as the diversified hydrophobic core of CD derivatives, the novel CD/CS nanoparticles developed herein might represent an interesting and versatile drug delivery platform for a variety of poorly water-soluble drugs with different physicochemical properties.