Objective To investigate the method and optimum conditions of electric transfection,and the major influential factors of electransfection efficiency and the survival rate of dendritic cells. Methods RNA was extracted from human hepatocarcinoma cell line(Bel 7402).Purified monocytes as precursor DC-s(pDC-s) were separated from human peripheral blood cells(PBMCs) by density gradient centrifugalization with lymphocyte gradation fluid and adherence method,pDCs were incubated in RPMI-1640 medium containing rhGM-CSF(8?10~5IU/L) and rIL-4(5?10~5IU/L) for 7 days and made them fully differentiate into immature DCs(imDCs).The total RNA human hepatocarcinoma cell and green fluorescent protein(GFP) were electransfected respectively into imDCs by electroporation apparatus with different electric voltages,times of impulse,cell concentrations,temperatures and electroporation buffers.Numbers of green fluorescence positive cells and the total cell number were counted respectively under fluorescent microscope,and visible light microscope.One day after the electric transfection,the cells were stained with 0.4% trypan blue,and electransfection efficiency and the cell survival rate were counted. Results Electransfection efficiency was increased to the highest value,up to about 49.7% when imDCs with the concentration of 5?10~6 cells/ml were mixed with 40?g-total RNA of human hepatocarcinoma cell,the electric voltage of electroporation apparatus was set at 300V,and the time of impulse was 500Us.Conclusion Electric transfection provides a technical possibility to make human hepatocarcinoma RNA into imDCs.The major influential factors of the electransfection efficiency were electric voltage and impulsing time.As receptor cells,the imDCs growing condition was also an important influential factor.

Objective:To probe the effect of noise on the capabilities of learning and memory of growing rat and its possible mechanism.Methods:24 Wistar growing rats were divided randomly into two groups,noise group and control group.The growing rats in the noise group were exposed to 90dB(A) noise constantly for one month.The capabilities of learning and memory of growing rats were measured by means of Morris water maze test.An immunohistochemical method was used to observe the expression of aminobuty-ric acid(GABA) positive neurons in hippocampus.Results:The test of Morris water maze showed that the mean escape latency of growing rats in the noise group were longer than that in the control group and the frequency to bestride platform were less.The number of GABA positive neurons and the magnitude of tinction in hippocampus of growing rats in the noise group were reduced markedly than that in the control group.Conclusion:The noise does impair the capabilities of learning and memory of growing rats.The impairment may be related with the reduction of GABA contents in hippocampus,which could decrease the depressive effect of neuronic synapse and impair the induction and maintenance of the long term potentiation.

Objective To observe the clinical effect of inhibition about gallstone stone after extracorporeal shock wave by Liver gallstones sheet. Methods 60 patients diagnosed gallstone stone from July 2015 to November 2016, randomly divided into treatment group and control group. he treatment group received oral treatment of liver gallstones sheet, control group without special treatment, patients were followed up and recorded gallbladder wall thickness, gallbladder function and gallstone patients were relapse situation, using statistical methods to analyze data. Results After one year observation of patients gallbladder wall thickness, gallbladder function scores were (2.79±0.34,57.70±4.80) than the control group (3.14±0.39,53.13±5.01), and the differences were statistically significant (P<0.05). After one year observation group gallstone recurrence rate was 10.0%, far lower than the control group, 43.3%, and the difference was statistically significant (P<0.05). Conclusion It is worthy of further research and application and it has a good clinical results for the liver gallstones sheet after extracorporeal shock wave lithotripsy inhibition of gallstone stone.

Objective To investigate the clinical effect of motor imagery therapy on upper limb motor dysfunction in stroke patients.Methods 120 patients with post stroke upper limb disability were selected.According to the digital table,they were divided into observation group and control group,60 cases in each group.Two groups were treated with primary disease and rehabilitation treatment,the observation group used motor imagery therapy,the two groups were treated for 3 months.Limb simplified Fugl-Meyer rating scale(FMA) and self-care ability Barthel index(MBI) were used to evaluate the motor function and self-care ability of the patients before and after treatment.The upper limb movement was evaluated by ARAT before and after treatment.The quality of life score before and after treatment was observed.Results Before treatment,there were no statistically significant differences in FMA and BI scores between the two groups(t=0.13,0.23,all P>0.05).After treatment,the FMA score of the control group was (29.74±4.04)points,the BI score was (57.29±4.23)points,the FMA and BI scores of the observation group was (33.29±4.14)points,(63.12±4.21)points,respectively,the differences were statistically significant between before and after treatment in the self-group(t=3.56,5.61,all P<0.05),and the degree of elevation of the observation group was more significant than those of the control group,the differences were statistically significant(t=2.38,4.14,all P<0.05).After treatment,the degrees of elevation of the scores of ARAT in the observation group[grab (13.34±2.00)points,grip (10.23±0.39)points,pinch (14.53±1.43)points,thick (8.20±0.42)points]were significantly higher than those in the control group,the differences were statistically significant(t=3.45,2.87,3.17,2.98,all P<0.05).After treatment,the rise degree of the scores of quality of life in the observation group[physiological field (68.71±6.62)points,psychological field (69.02±6.38)points,social relations (67.72±6.79)points,environmental field (70.72±6.98)points]were higher than those in the control group,the differences were statistically significant(t=4.23,3.98,3.23,2.98,all P<0.05).Conclusion The effect of exercise therapy on the upper limb of patients after stroke has significantly clinical efficacy,and it can help patients to restore limb motor function,improve self-care ability and quality of life,it is safe and worthy of clinical promotion.

al blood lactic acid monitoring is simple, easy and practical to use in predicting prognosis of ICU patients sensitively.

Objective To explore the relationship between epilepsy and stroke,the features of secondary epilepsy and its treatment. Methods 233 stroke patients were observed and the clinical data of 21 patients who had secondary epilepsy were analyzed. Results Secondary epilepsy after stroke was mainly seen in the cortical stoke patients, the incidence of epilepsy of the cortical stroke group was 16.7% , which was significantly higher than that of the gubcortical stroke patients(5.7% ) ,and there was significant difference between the two groups. The incidence of epilepsy of stroke with a lesion >4cm group was 17. 3% .which was significantly higher than that of stroke with a lesion <4cm group(5.4% ), there was also significant difference between the two groups. There was no significant difference between ischeraic stroke and hemorrhagic stroke(P>0.05). Conclusion The incidence of secondary epilepsy after stroke had no correlations with the stroke types,but was related to the area and the location of the stroke.

Objective: A method for the determination of gallic acid in the extract of Quercus salece- na Blume (Fagaceae) and its preparation (Urocalum Capsules) by means of RP-HPLC was established. Method: The hydrolysable tannins contained in the sample is hydrolyzed with hy- drochloric acid, the hydrolysis component-gallic acid is determined by HPLC using RP-C18 column, 0.005mol. L-1 of phosphoric acid-methanol (98: 2) as mobile phase , 270 nm as detec- tion wavelength. Result: The linear range of gallic acid is 0. 3～3. 1?g , r=0. 9999, the average recovery is 100. 4%. Conclusion: It is confirmed that the established method for assay of gallic acid meet the criteria of quantitative analysis of herbal medicine by validation of hydrolysis condition, repeatebility, stability and recovery, The method is simple , good reproducible and precise , and it can also be applied to analysis of other herbal medicine containing hydrolysable tannins.

AIM:To study effects of resveratrol on osteoprotegerin(OPG)and receptor activator of NF-?B ligand(RANKL)expression in femur of ovariectomized rats.METHODS:48 female SD rats(3 months old)were assigned into 6 groups randomly:sham-operated group(SHAM);ovariectomized group(OVX);17?-estradiol replacement therapy group(ERT,0.1 mg?kg-1?d-1,sc);high-dose resveratrol-treate group(RH,40 mg?kg-1?d-1,i.p.);middle-dose resveratrol-treated group(RM,20 mg?kg-1?d-1,i.p.);low-dose resveratrol-treated group(RL,10 mg?kg-1?d-1,i.p.).Except for SHAM group,rats in other groups were operated by bilateral ovariectomy.Administration started at the 8th day after operation and lasted 8 weeks.At the end of the 9th week after operation,all rats were sacrificed.The bone mineral density(BMD)and vitodynamics properties(including ELASTIC,M-STRESS,M-LORD and STIFFNESS)of femurs were measured.The expression of OPG and RANKL in femurs of rats were observed by immunohistochemical staining methods.RESULTS:Compared with OVX group,resveratrol treatment(40 mg?kg-1?d-1 and 20 mg?kg-1?d-1)increased expression of OPG in femurs while decreased expression of RANKL in ovariectmized rats.In RH,RM,and ERT group,BMD,ELASTIC,M-STRESS of femurs were higher than those of OVX group.CONCLUSION:Resveratrol increased expression of OPG in femur while decreased expression of RANKL in ovariectmized rats,which was probably one of mechanism that resveratrol improve osteoporosis.

Objective To investigate the effect of nuclear factor-?B (NF-?B), NF-?B-inducing kinase (NIK) and I-?B in basic fibroblast growth factor (bFGF) promoting gene expression and signal transduction of tenocyte NF-?B. Methods Tenocytes isolated from rabbits were divided into 3 groups with 8 dishes: no bFGF (Group A), 2 ng/ml bFGF (Group B) and 10 ng/ml bFGF (Group C). Cells in all dishes were cultured for 5 days. The effect of bFGF on tenocytes proliferation was examined by means of growth curve analysis. The mRNA was isolated after 5 days and the gene expressions of NIK,IKK? and IKK? induced by NF-?B investigated by reverse transcription-polymerase chain reaction (RT-PCR). Results The growth curve moved forward with the dosage increase of bFGF. Gene expressions of NF-?B,NIK, I?B kinase (IKK)? and IKK? were 0.4? 0.2 , 0.4 ? 0.1 , 0.3 ?0.1 and 0.2?0.1 respectively in the Group B and 1.8?0.5,1.0?0.3, 0.8 ? 0.2 and 1.5 ? 0.4 in the Group C. Gene expressions were all up-regulated in the Groups B and C posterior to adding of bFGF. Conclusions bFGF stimulates tenocyte proliferation and up-regulates gene expression of NF-?B,NIK,IKK? and IKK?, as indicates that the signal transduction of promoting tenocyte proliferation by bFGF may be through NF-?B pathway.