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.

Child ; Humans ; Neoplasms

Purpose To assess the potentia1 of DNA image cytometry and urinary cyto1ogy in screening for urothe1ia1 carcinomas,and to find the idea1 method of ear1y diagnosis of urothe1ia1 cancer. Methods Voided urine specimens from 121 patients with urothe1ia1 carcinoma were random1y co11ected as test group and 95 patients with symptomatic uro1ogic disease but not urothe1ia1 carcinoma as con-tro1 group,a11 of the specimens were ana1yzed by both urinary cyto1ogy and DNA image cytometry. Results Urinary cyto1ogy had a higher sensitivity but 1ower specificity in diagnosis of urothe1ia1 carcinoma than that of DNA cytometry. Urinary cyto1ogy yie1ded an o-vera11 sensitivity of 85. 10%,specificity of 76. 80%. DNA cytometry revea1ed a sensitivity of 81. 80%,specificity of 81. 10%. Among the 121 urothe1ia1 carcinoma,there were 103 b1adder cancers,18 patients with upper urothe1ia1 carcinoma. For different types of urothe1ia1 carcinoma,both of the two methods demonstrated good sensitivity in the high 1eve1 tumor. And DNA cytometry had exce11ent sensitivity in the diagnosis of urothe1ia1 carcinoma in invasive b1adder cancer and upper urinary tract cancer,and both were more than 94%. Conclusions In 1ight of its high1y good sensitivity and specificity in urothe1ia1 carcinoma,especia11y in invasive b1adder cancer and upper urinary tract cancer,DNA cytometry shou1d be used to eva1uate suspect urothe1ia1 ce11s in urinary cyto1ogy specimens. Com-bined with the conventiona1 urinary cyto1ogy,the detection for urothe1ia1 carcinoma wou1d be significant1y improved.

Delayed cerebral vasospasm is the leading cause of death in patients with subarachnoid hemorrhage. A good in vivo animal model of cerebral vasospasm after subarachnoid hemorrhage helps to understand the occurrence, development of this cerebral vasospasm, and the complicated and various pathophysiological mechanisms. It will provide important information for clinical interventions

Periodontal ligament stem cells(PDLSCs) are not only the crucial type of cells in maintaining dynamic homeostasis and wound repair of periodontal tissues but also the basis for novel therapeutic strategy for periodontal regeneration.Although possessing the excellent proliferation and differentiation ability,PDLSCs behave differently under various conditions representative of several influential factors.Understanding of these influential factors will help us to exploit more about PDLSCs,and more importantly,lead a new modality in treating periodontal diseases based on stem cells.Here we review multiple important factors affecting function of PDLSCs and propose its use in future regenerative therapy of periodontal defects.

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.