This paper is to report our study of using single radical hemolysis(SRH) technique to detect the specific antibody for Japanese B encephalitis (JBE) virus in 101 samples of pigs' serum at Chongqing area. It was found that SRH was more sensitive and more specific in the detection of the JBE virus antibody in the pig's serum than CF or HI. SRH is simple in its technique and easy to perform. In addition, it is very sensitive and specific and it can be reproduced easily. It is suggested that SRH be used in clinical diagnosis and in seroepidermic survey of JBE virsus infection.

Objective: To evaluate the characteristic and explore the mechanism of microwave extraction (MAE) in extracting Chinese medicines by comparing with conventional extractions on Radix et Rhizoma Rhei. Methods : The sum of anthraquinone was determined by spectrophotometry and aphrostase in paraffin section was observed by microphotography. Results : Among the four methods, the efficiency of MAE was significantly the highest, which was 3.5 times of supersonic extraction and 1.5 times of Sohlex extraction and 1.5 times of decocting by water, respectively. The time of MAE was the shortest. MAE could destroy the cell organization to pick up the speed of dissolving. Conclusion : MAE is efficient, saving energy and time in extracting Chinese medicines.

AIM: To evaluate the characteristic and explore the mechanism of MAE on Semen Cassiae by comparing MAE with commonly used extraction method. METHODS : The amount of anthraquinone was determined by spectrophotometer. The surface and cross section of Semen Cassiae were observed by microphotography. RESULTS : Among the four methods,the efficiency of MAE is 16 times that of ultrasonic extraction,3 times that of Sohlex extraction and 1.1 times of decocting by water,respectively. Micrographs taken after extraction differed markedly indicated that the degree of damage varied considerably. CONCLUSION : The MAE method is more advantageous than other traditional extraction methods (Soxhlet extraction and ultrasonic extraction) with regard to the extraction yield and the time and cost of the procedure.

Objective: To establish a fluorescence method based on turncated aptamer for the determination of bisphenol A in water. Methods: The bisphenol A truncated aptamer containing 38 bases was selected as a recognition module, and was modified with the fluorophore 6-FAM at the 5'end. The 3'end of the complementary sequence cDNA was modified with the quencher DABCYL. The standard solutions of bisphenol A and interfering compounds were configured. The detection system was established after optimizing the number of bases in cDNA, the concentration ratio of truncated aptamer to cDNA, the incubation temperature and time, and the pH of the buffer. The specificity and recovery experiments were carried out. Results: When the complementary sequence cDNA included 9 bases, the concentration ratio of the truncated aptamer to cDNA was 1:1.5, the pH value of the buffer solution was 7.5, the cDNA was incubated at 55 ℃ for 60 minutes, in the concentration range of 10-75 pmol/L, the linear regression equation was y=2 230.7x+110 825, the correlation coefficient was 0.926. The limits of detection was 3.3 pmol/L. The difference values of fluorescence intensity between tetrabromobisphenol A, estradiol, estriol, bisphenol S and bisphenol A were obviously different, so there was no significant interference to the test result. The recovery rates were 97.8%, 98.8% and 102.3% with the spiked concentrations of 20.0, 40.0 and 60.0 pmol/L. The relative standard deviations were 4.4%, 2.1% and 2.6% (n=5), respectively. Conclusion The fluorescence method based on turncated aptamer has the advantages of easy operation, high sensitivity and specificity, which can be used for the determination of bisphenol A in water.

Object To show that ELSD is an excellent detector for the detection of chemical compounds devoid of chromophore, such as sarsasapogenin Methods Sarsasapogenin in crude Anemar rhena asphodeloides Bunge and its preparation was determinated by RP HPLC ELSD Results The well separated chromatographic peaks show linearity with recovery of the added sample of 100 5% in crude medicinal material and 91 38% in its preparation, r= 0 999 0 Conclusion The method was advanced, reliable, simple and can be used for quality control of crude A asphodeloides and its preparations

Object To explore the regularity of microwave extraction (ME) on Chinese medicines in different morphological structure and different polar compositions. Methods Anthraquinone in Radix et Rhizoma Rhei (RRR), Semen Cassiae (SC), cholorogenic acid in Flos Lonicerae, baicalin in Radix Scutellariae were determined as index compositions by HPLC. The extraction rate was measured by orthogonal design. Results ME selectivity to different anthraquinone in RRR is not significant, while at the same temperature, the extraction rates of emodin, chrysophanol, physcion in RRR are higher than those in SC. Conclusion The ME selectivity to the different morphological structure of Chinese medicines is obvious, but to the different polar compositions is not distinct.

Objective To establish a method for isolation of cynomolgus monkey umbilical cord mesenchymal stem cells.Methods Fresh cynomolgus monkey umbilical cord was directly minced into pasty fine pieces, and the pieces were cultured in tissue flask with DMEM/F12 medium supplemented with 10% fetal bovine serum.The morphological characteristics of the resulting cells were examined, and their expression of mesenchymal cell surface markers were analyzed by flow cytometry.The multidifferentiation potential was examined in vitro, too.Results The fibroblast-like cells were successfully isolated from the fresh umbilical cord by an adherent culture procedure.These adherent cells expressed mesenchymal markers including CD29, CD44, and CD90, and also could be induced to differentiate into adipocytes, osteoblasts and chondrocytes.Conclusion Mesenchymal stem cells can be isolated from fresh cynomolgus monkey umbilical cord by using an adherent culture procedure.

Objective To investigate the influences of grape seed proanthocyanidin extract (GSPE) on cardiovascular system, nervous system and respiratory system of experimental animals, and provide general pharmacological data for further research and application. Methods The influences of GSPE on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs after duodenal administration were observed, the impacts on spontaneous activity, coordinated motion, and the sleep situation with threshold dose and subthreshold dose of pentobarbital sodium in mice after intragastric administration were observed. Results GSPE showed no side effects on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs at the dosage of 857.00, 214.29, 42.86 mg/kg (P>0.05). At the dosage of 428.57, 214.29, 42.86 mg/kg, GSPE had no obvious influence on spontaneous activities and coordinated movements in mice (P>0.05). GSPE did not evidently change the number of sleeping animals, the sleep latency and the sleeping duration with subthreshold dose and threshold dose of pentobarbital sodium (P>0.05). Conclusion GSPE has no evident adverse effects on central nervous system, cardiovascular system and respiratory system in animals.

BACKGROUND: Studies have shown that umbilical cord mesenchymal stem cells are ideal seed cells for tissueengineering research.OBJECTIVE: To isolate, culture and identify tree shrew umbilical cord mesenchymal stem cells, in order toestablish a standardized tree shrew umbilical cord mesenchymal stem cell lines.METHODS: Caesarean-isolated tree shrew umbilical cord samples were used to isolate and culture umbilical cordmesenchymal stem cells using tissue explant adherent method. Flow cytometry assay was used to detect cellsurface markers. Osteogenic and adipogenic induction media were used to induce umbilical cord mesenchymalstem cells to differentiate into osteoblasts and adipocytes.RESULTS AND CONCLUSION: The cultured umbilical cord mesenchymal stem cells expressed CD90 and CD105 with the positive rate of 99.9% and 99.8% respectively. Hematopoietic stem cell marker CD34 expression ratewas 0.0% and the endothelial cell marker CD31 expression rate was 0.7%, in line with the characteristics of umbilicalcord mesenchymal stem cell surface markers. Calcium nodules by alizarin red staining and lipid droplets by oil red Ostaining were observed in the induced cells. These experimental findings indicated that umbilical cord mesenchymalstem cells from tree shrews capable of osteogenic and adipogenic differentiation were successfully isolated and cultured.

AIM:To explore the effects of confluent endothelial progenitor cells (EPCs) derived from young and aged rats on the phenotype conversion, proliferation and migration of vascular smooth muscle cells ( SMCs) .METH-ODS:Mononuclear cells were obtained from the bone marrow of young (1~2 month old) and aged (19 to 26 month old) Sprague-Dawley rats and cultured with medium DMEM/F12 ( containing 15% fetal bovine serum, endothelial cell growth supplements (ECGs) 100 g/L, 1 ×105 units/L of penicillin and streptomycin, respectively).EPCs were characterized as double positive for DiI-Ac-LDL uptake and lectin binding.Abdominal aorta was obtained from 1 to 2 month old Sprague-Dawley rats.Vascular SMCs were cultured by tissue explant method and identified byα-SM-actin immunofluorescence.In transwell co-culture system, the confluent EPCs located in the upper chamber and SMCs were seeded on the lower cham-ber.The experiments were divided into passage 3 SMCs group (P3), passage 4 SMCs group (P4), passage 4 SMCs co-culture with EPCs derived from young rats group (P4YE) and passage 4 SMCs co-culture with EPCs derived from aged rats group (P4AE).The protein expression ofα-SM-actin and osteopontin was detected by Western blotting.[3H]-TdR incor-poration assay was used to determine the proliferation.SMC migration was analyzed by scratch wound healing assay.RE-SULTS:Compared with P3 group,α-SM-actin expression in P4 group significantly decreased and osteopontin protein ex-pression obviously increased, whereas no significant change was found in P4YE group.Compared with P4 group, confluent EPCs derived from young and aged rats both markedly increased α-SM-actin and decreased osteopontin expression in P4 SMCs.Compared with aged rat-derived EPCs, young rat-derived EPCs were more effectively to induce a delayed SMC phe-notype transition (from contractile phenotype to a synthetic phenotype), and to inhibit SMC proliferation and migration. CONCLUSION:Co-culture of confluent EPC induces a delayed vascular SMC phenotype transition and inhibits SMCs pro-liferation and migration.Young rat derived EPCs are more effective to induce a delayed vascular SMC phenotype transition and has stronger inhibitory effects on SMCs proliferation and migration compared with that derived from aged rats.