BACKGROUND: The study indicated that directed neural differentiation of bone marrow stromal cells (BMSCs) was associated with changes in local micro-environment in injured spinal cord following cerebral ischemia, in particular neurotrophic factor induction. Group pre-test has confirmed that acupuncture can increase the expression of vadous cytokines and neurotrophic factors to promote nerve regeneration and repair.OBJECTIVE: To observe the effects of electro-acupuncture combined with BMSC kansplantaUon on recovery of neurological function in rats with spinal cord injury.DESIGN, TIME AND SETTING: Randomized controlled animal experiments were performed at the Laboratory of Cell Biology,Harbin Medical University from March 2005 to July 2006.MATERIALS: A total of 80 pure healthy Sprague Dawley rats were selected. Eight were used for the isolation and culture of BMSCs, and the remaining 72 rats were equally and randomly divided into 4 groups: blank control group, cell transplantation group, electro-acupuncture group, and combination group. KWD-80811 pulse-type apparatus was produced by the third radio factory in Jiangsu Wujin production.METHODS: In vitro isolation and culture of the third passage of BMSCs were obtained and labeled by BrdU at hour 72 before transplantation, and BMSCs at 1 ×10 11/L were harvested for usa. Rat models of spinal cord injury were established in each group.Following model establishment, BMSC suspension was injected into the injured spinal cord near the junction of the gray matter and white matter in the cell transplantation group, totally 6 × 10<'5> cells. The same volume of phosphate buffer saline was infused into rats of the blank control group by the same method. In the electro-acupuncture group, at hour 24 after the successful model establishment, Jiaji Electroacupuncture Electroacupuncture instrument treatment was performed by pulse electro-acupuncture apparatus at 3.0-4.0 mm from median line in the upper and lower end of the spinous process vertebral body adjacent to open space for 20 minutes, once a day. In the combination group, rats underwent BMSC transplantation + Jiaji electro-acupuncture treatment. MAIN OUTCOME MEASURES: After transplantation for 3, 7, 14 days, neurological conditions were assessed by combined behavioral scores in rats with spinal cord injury. Glial fibrillary acidic protein and neuron-specific enolase expression was measured in BrdU-labeled BMSCs by immuno-double-labeling method.RESULTS: 3, 7, 14 days after injury, compared with the blank control group, significant differences in combined behavioral scores were determined in the call transplantation group, electro-acupuncture group, and combination group (P <0.05, 0.05, 0.01).The recovery of neurological function was significantly greater in the combination group than in the cell transplantation and electro-acupuncture groups (P < 0.05). No significant difference was detected between the cell transplantation group and the electro-acupunctura group (P > 0.05). Compared with blank control group, spinal cord structure was relatively complete in the cell transplantation group and electro-acupuncture group. The structure was more integrity in the combination group. Bone marrow stromal cell transplantation presented the organization BrdU-labeled cells in the lesion and the surrounding area, with obvious aggregation and survival. At day 14 following cell transplantation, positive rates of neuron-specific enolase and glial fibrillary acidic protein were respectively 7.2% and 1.5% in the call transplantation group, 7.9% and 2.1% in the combination group.CONCLUSION: The BMSCs after transplantation can survive in the host body. Electro-acupuncture could promote the differentiation of BMSCs into neural cells. Combination of electro-acupuncture and BMSC transplantation can significantly improve the motor and sensory function in rats with spinal cord injury.

OBJECTIVE：To investigate the activity of lycorine to the in vivo apoptosis of tumor cells in H 22-bearing mice and its mechanism. METHODS ：Kunming mice were inoculated subcutaneously with ascites of H 22 hepatoma mice in the armpit of forelimb to establish solid tumor model. After modeling ，mice were randomly divided into negative control group ，positive control group（hydroxycamptothecin 6 mg/kg），lycorine low-dose ，medium-dose and high-dose groups （10，20，40 mg/kg），with 10 mice in each group. Negative control group was given constant volume of normal saline intragastrically ，and administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 7 days. After last medication ，the weight of tumor was detected and anti-tumor rate was calculated. Ascites tumor model of mice was established by intraperitoneal injection of H 22 hepatoma mice ascites ，and then were grouped with same method and given relevant medicine as above. After last medication ， survival time of mice was recorded and the life prolongation rate was calculated. The early apoptotic rate of tumor cells in mice was detected by flow cytometry. On the basis of normal control group （normal mice without tumor ），the mitochondrial membrane permeability of tumor cells in each group was investigated by Calcein AM staining. The changes of mitochondrial potential were investigated by Rhodamine 123 staining. Colorimetry and Western blot assay were adopted to detect the Caspase-3 activity and expression of apoptosis-related protein （Bcl-2，Bax，Cyt-C and Caspase- 9）. RESULTS ：Compared with negative control UN- group，the tumor weight of positive control group and lycorine PYSCT-2017208） groups were decreased significantly ，while the survival time was significantly prolonged ，and the early apoptotic rate of tumor cells was significantly increased （P＜0.05 or P＜0.01）；the anti-tumor rates were 39.41％ ， 23.36％ ， 36.50％ ， 56.93％，and life prolonga tion rates were 49.23％，29.09％， E-mail：ym913@yahoo.com.cn 50.19％，69.08％. Compared with normal control group ，the mitochondrial membrane permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased，while the mitochondrial membrane potential and Bcl- 2/Bax ratio were decreased significantly （P＜0.05 or P＜0.01）. Compared with negative control group ，mitochondrial membrane permeability and Bcl- 2/Bax ratio were decreased significantly in administration groups ，while mitochondrial permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Lycorine can induce the apoptosis of tumor cells in H22-bearing mice ，the effects of which may be associated with opening mitochondrial membrane permeability transition pore to increase mitochondrial permeability ， decreasing mitochondrial membrane potential and up-regulating the expression of apoptosis-related proteins.