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

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.

BACKGROUND:Umbilical cord mesenchymal stem cels are from the umbilical cord of newly born individuals and have no ethical issues, and therefore are promising candidates for seeded cels as a substitute for cel transplantation and regenerative medicine. OBJECTIVE:To investigate the effects of serum from liver injury rats on induced differentiation of human umbilical cord mesenchymal stem cels into hepatocyte-like cels and provide experimental evidence for use of human umbilical cord mesenchymal stem cels in the treatment of patients with end-stage liver disease in the clinic. METHODS: Rat models of acute liver injury were established by intraperitoneal injection of 10% carbon tetrachloride. Rats in the control group were intraperitonealy administered the same amount of soybean oil. Forty-eight hours after modeling, abdominal aorta blood was taken for serum preparation. Passage 3 human umbilical cord mesenchymal stem cels were cultured with 20% serum from liver injury rats and 20% fetal bovine serum. Morphology of human umbilical cord mesenchymal stem cels was observed before and after culture. Levels ofα-fetoprotein and albumin in the supernatant were detected. RESULTS AND CONCLUSION:Cels exhibited shuttle-shaped appearance and grew in whirlpool-like manner at 1 day after culture with serum from liver injury rats, exhibited short shuttle-shaped appearance at 2 days, were oval-shaped at 3 days, and were round and an extremely smal number of cels were floated at 4 days. At 4 days after culture with serum from liver injury rats, level of albumin in the cel supernatant was significantly increased than that before induction and that in the control group (P< 0.001), and there was no significant difference in level of α-fetoprotein in the cel supernatant. These results suggest that serum of liver injury rats can induce differentiation of umbilical cord mesenchymal stem cels into hepatocyte-like cels.

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.

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.

OBJECTIVE: To investigate and statistics the application of the oral L-carnitine in the outpatient departments of our hospital, and to promote clinically rational drug use. METHODS: A total of prescriptions were collected from the information system in our hospital during 2010-2012, then the software of excel was employed to statistically analyze the data. RESULTS: The application of the oral L-carnitine in urology was the most widely, followed by the department of neurology, gastroenterology, medical and pediatrics. The oral L-carnitine was mainly used to treat the disease of male infertility, varicocele, male hypogonadism, metabolic myopathy, mitochondrial disease, fatty liver, organic acidemia and other diseases. CONCLUSION: The oral L-carnitine is widely used to treat the diseases which induced by carnitine deficiency. The off-label indications of the oral L-carnitine existed in the department of urology, so the safety and effectiveness should be concerned.

Objective: To predict the growth trend of total health expenditure in China based on the dynamic relationship between total health expenditure and economic growth. Methods: First, analyze the relationship between total health expenditure and economic growth from 1978 to 2011 with the state space model, evaluate the accuracy of the model and predict total health expenditure range from 2012 to 2016 with the status space mode. Results: Up to 2016, the total health expenditure would reach to 5.681 trillion yuan, and the ratio of the expenditure to gross domestic product(GDP) would increase to 5.692%. Conclusion: It is needed to balance the relationship between total health expenditure and economic growth.