[Objective] Ursolic acid and oleanolic acid in Acantopanax senticosus (Rupr. et Maxim.) Harms were extracted with the ultrasonic technology. A determination method of ursolic acid and oleanolic acid was established with the high performance liquid chromatograph and photodiode array detector. [Method]The chromatographic column, Nova-Pak C18 (3.9?300 mm, 4 ?m), methanol-water (88∶12,V/V), mobile phase with 0.8 mL/min flow rate, the detected wave length (210nm),and the column temperature(25 ℃) were adopted. [Results] The calibration curve of ursolic acid was linear under the content of 0.136 ~1.224 ?g and the correlation coefficient was 0.9995. The calibration curve of oleanolic acid was linear under the content of 0.052 ~0.468 ?g and the correlation coefficient was 0.9994. The average recovery of ursolic acid was 98.1 % and RSD was 1.1 % (n=5). The average recovery of oleanolic acid was 97.5 % and RSD was 1.0 % (n=5). [Conclusion]The content determination of HPLC is accurate and reliable, and can be used to analyse quantificationally ursolic acid and oleanolic acid in Acantopanax senticosus (Rupr. et Maxim.) Harms.

To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.
Agrobacterium ; Chromatography, Thin Layer ; Culture Media ; Dianthus ; growth & development ; Plant Growth Regulators ; Plant Leaves ; Plant Roots ; growth & development ; Plants, Genetically Modified ; Rhizobium ; Tissue Culture Techniques ; Transformation, Genetic

Objective:To explore the effect of mind map on health education of children with asthma and their parents.Methods:Ninety children with asthma admitted for the first time were divided into observation group (45 cases) and control group (45 cases) by random digits table method. The control group used the traditional health education method, and the observation group used the mind map pattern health education method. The drug compliance of the two groups was evaluated before discharge and at the 1st, 2nd,3rd month after discharge, and the satisfaction of the children and parents and therapeutic effect of the two groups were compared at the time of discharge.Results:There was no significant difference in treatment adherence between the two groups before discharge ( P > 0.05). The treatment adherence at the 1st, 2nd,3rd month after discharge was (7.22±1.31), (7.58±1.55), (7.78±2.11) points in the observation group, and (6.98±2.01), (7.01±1.58), (7.11±2.48) points in the control group, there were significant differences between the two groups ( t values were 4.426, 3.884, 5.233, all P < 0.05). The satisfaction of the children and parents was (96.01±3.45) points in the observation group,and (78.13±2.39) points in the control group, there was significant difference between the two groups ( t value was 38.223, P < 0.01). The total effective rate was 77.8%(35/45) in the observation group,and 64.4%(29/45) in the control group, there was significant difference between the two groups ( χ2 value was 4.013, P < 0.05). Conclusions:Health education integrated with mind map can effectively improve the satisfaction of children and their parents, and improve the drug adherence and therapeutic effect of children.

To use hairy roots for producing medicinal ingredients of Phytolacca americana L. we studied the factors influencing the induction and in vitro culture. Hairy roots could be incited from the veins of cut surface (morphological lower) of P. americana L. leaf explants around 18 days after infection with the strain of Agrobacterium rhizogenes ATCC15834. The highest rooting rate, 70%, was obtained when leaf explants were pre-cultured for 1 day, infected for 20 min, and co-cultured for 4 days. The transformation was confirmed by PCR amplification of rolC of Ri plasmid and silica gel thin-layer chromatography of opines from P. americana L. hairy roots. All the hairy root lines could grow rapidly on solid exogenous phytohormone-free MS medium. Among the 9 hairy root lines, the hairy root line 2 had most rapid growth, most branched lateral roots and most intensive root hair; the root surface of some hairy root lines seemed purple or red, while that of the other hairy root line appeared white. Among liquid media MS, 1/2MS, B5 and 6,7-V tested, the best growth for hairy root lines was attained in liquid exogenous phytohormone-free MS medium. Compared with exogenous phytohormone-free MS medium, 6,7-V medium was better for synthesis and accumulation of esculento side A in hairy roots. The established optimal conditions for induction and in vitro culture of P. americana hairy roots had laid an experimental and technological foundation for production of medicinal constituents esculento side A from large scale culture of hairy roots.