Objective: To investigate the interaction between tanshinone IIA (TS IIA) and cholesteryl ester transfer protein (CETP), and to explore the ways of impact on CETP. Methods: The various structures of TS IIA and CETP were built based on the crystal structure and then performed molecular dynamics (MD). The simulation software is Gromacs 4.0 with force field of Gromos 96 53a6. The temperature is 300 K and the simulation time is 20 ns. All trajectories were recorded to analyze the changes of overall shape and local structures of CETP, and the interaction energy between TS IIA and CETP. Results: When the phosphatidyl choline zones of CETP were full, the structure was rigid. When only the cavity was loaded, CETP was easy to change. The out area of two side, phosphatidyl choline area and cavity change greatly corresponded to the frame changes of CETP. Stronger interaction between TS IIA and CETP occured in the two phosphatidyl choline area and the left side of cavity. Conclusion: CETP is a carrier protein with easily changed structure which changes according to the differences in the number and structures of loading ligands. TS IIA might affect the morphology of the CETP and then inhibit its transportation ability under the help of phosphatidyl choline or cholesterol ester.

Objective: To construct the eukaryotic expression vector of human survivin gene and stably transfect it into mouse melanoma B16 cell line. Methods: The full length human survivin cDNA fragment was amplified by PCR and inserted into eukaryotic expression vector pIRES-neo; the restriction enzyme position and 6 his tag were added to obtain recombinant plasmid pIRES-neo-SUR-(his) 6, which was then transfected into B16 cells by Lipofectamine 2000. After screening culture by G418, a stably transfected cell line was established, and the transcription and expression of the human survivin gene were identified by RT-PCR, Western blotting analysis and immunofluorescence assay. Results: The result of restriction enzyme digestion and the sequence analysis showed that the recombinant of pIRES-neo-SUR-(his)6 was successfully constructed. RT-PCR results showed survivin gene (about 530 bp) was amplified from the total RNA in the group stably transfected with pIRES-neo-SUR-(his)6. Western blotting analysis showed the expression of survivin protein in pIRES-neo-SUR-(his)6 transfection group, but not in the control group. FACS and immunofluorescence assay showed high fluorescence signal in pIRES-neo-SUR-(his)6 transfection group, with the fluorescence positive rate being 91.38% when No. 5 monoclonal antibody was used; no fluorescence signal was found in the control group( transfected with pIRES-neo). Conclusion: We have successfully constructed the eukaryotic expression vector of human survivin pIRES-neo-SUR-(his)6, and established a B16 cell line stably and highly expressing human survivin gene.

The immunoprotective effect of Bacille Calmette-Guerin (BCG) is controversial. Appearance of multi-drug resistant tubercle bacilli(MDR-TB) and prevalence of AIDS lead to upswing of tuberculosis (TB) incidence, which poses a challenge for TB control. Therefore, the new TB vaccines become an urgent need for prevention and treatment of TB. This article will review the latest developments of TB vaccine research.

Objective: To study the culture condition and biological characteristics of canine multipotent adult progenitor cells(MAPC) and their potential to differentiate into smooth muscle-like cells in vitro. Methods: The morphology and growth of in vitro cultured MAPC were observed and the surface marker of MAPC was detected. The cell cycle and phenotype of MAPC were determined by FACS. RT-PCR was used to examine the expression of OCT-4 in MAPC and smooth muscle cells-myosin heavy chain (SM-MHC) in the differentiated cells. The surface markers of smooth muscle cells were detected in the differentiated cells. Results: Under microscope, MAPCs were polygon and had large nuclei, multi-parapodiums and affluent cytoplasma. The cells also showed strong proliferation ability and were positive of CD13, SSEA-1 and negative of CD44, CD45, CD133, and MHC II. RT-PCR showed expression of OCT-4. The cells differentiated from MAPC expressed the surface markers of smooth muscle cells, including α-SMA, calponin and SM-MHC. RT-PCR demonstrated the expression of SM-MHC. Conclusion: We have successfully cultured canine MAPC in vitro, which has similar biological characteristics as reported previoulsy. MAPC has the potential to differentiate into smooth muscle-like cells under certain physiological condition in vitro.

OBJECTIVE: To explore the proliferation inhibition and apoptosis induction effect of dextran-magnetic layered double hydroxide-fluorouracil (DMF) drug delivery system on colon cancer cell SW480 cultivated in vitro. METHODS: MTT experiment was used to detect the cell proliferation inhibition rates of DET-MLDH-FU, MLDH-FU and MLDH supra-molecules at various concentrations. Light microscopy and Giemsa dyeing methods were applied for characterizing the shape change of the apoptotic cells. Flow cytometric analysis was employed to test the apoptosis rates of SW480 cells treated with different drugs for 24 h. DNA ladder method was used to analyze nuclei breakup of the apoptosis cells. RESULTS: The IC50 of MLDH, MLDH-FU and DMF three-level supra-molecules for SW480 cells were 44.83(25.74, 48.78), 15.16(13.78, 16.66) and 13.33 (11.03, 15.13) μg · mL-1, respectively. The proliferation of colon cancer cells SW480 was obviously restrained after being treated with 5-Fu and equivalent MLDH, MLDH-FU and DMF of different concentrations for 24, 48 and 72 h, respectively. The result revealed that the inhibition rates increased in the sequence of MLDH

OBJECTIVE: To study the effect of AICAR on uterine smooth muscle of pregnant and nonpregnant rats. METHODS: The action of AICAR and dibutyryl-cAMP on the contractile function of isolated myometrium of pregnant and nonpregnant Sprague-Dawley rats was investigated. AICAR or dibutyryl-cAMP was used at cumulative doses from 10-9 to 10-4 mol · L-1 for myometrium of pregnant rats, and at 10-5 mol · L for myometrium of nonpregnant rats. The amplitude of the contractility of myometruim was recorded. RESULTS: For the myometrium of pregnant or nonpregnant rats, AICAR and dibutyryl-cAMP exerted strong inhibitory action on the contractile function in dose-dependent manners. For the myometrium of pregnant rats, AICAR or dibutyryl-cAMP showed inhibitory effect at a low concentration of 10-7 mol · L-1. AICAR and dibutyryl-cAMP also caused significant inhibition in myometrium from nonpregnant rats. CONCLUSION: AICAR exerted strong inhibitory action on the contractile function of myometrium from pregnant or nonpregnant rats, suggesting that AICAR may have inhibitory effect on spontaneous activity of myometrium of human.

Objective: To analyze the genetic relationship among the medicinal plants of Potentilla L. Methods: Six species of germplasm resources in Potentilla L. were analyzed by ISSR molecular markers. To make up the systematic diagram of genetic relationship by Popgene 1.32 software, cluster by UPGMA method, and establish the dendrogram. Results: A total of 105 ISSR bands were scored for 11 primers, among which 89 were polymorphic bands. The average percentage of polymorphic bands was 84.76%. Genetic similarity coefficient changed from 0.4476 to 0.7905. Conclusion: By cluster analysis, it shows that some of the medicinal plants in Potentilla L. from the same region are in the same group and demonstrates the rule of geographical distribution in the tested materials. This study will provide the foundation for identification and development of medicinal plants in Potentilla L., and guide collection and evaluation of germplasm resources.

Objective To develop a simple, rapid and accurate analysis method for determination of L-corydalmine (L-CDL) in rat plasma. Methods The plasma sample was determined with propranolol as the internal standard (IS), the separation was accomplished in a Capcell PAK C18(2.1 mm×750 mm, 3 μm) column, and the mobile phase consisted of water (including 0.1% formic acid and 5 mmol/L ammonium formate) and acetonitrile (including 0.1% formic acid) at a flow rate of 0.25 ml/min. The mass spectrometer consisted of an ESI interface operating at positive ionization mode and the detection was performed using multiple reaction monitoring(MRM) at the transitions of m/z 342→192.1 for L-CDL and m/z 260→116.2 for propranolol. Method validation included the evaluation of the linearity range, lower LOQ, within-run and between-run precision, accuracy and stability, matrix effect and extraction recovery. Results The calibration curve was linear across the concentration range of 1-5000 ng/ml for L-CDL with a lower LOQ of 1 ng/ml. The within-run and between-run precision (RSD%) was in the range 0.1%-10%. The extraction recovery was in 93.5%-102.8% and the matrix effect for three QC was 108.3%-112.5%. L-CDL reached the peak concentration at 0.5 h after dosing. The main pharmacokinetic parameters of rats after igl administration were as follows: T1/2: (7.04±3.93)h, Cmax: (557.8±330.1)ng/ml, AUC0~24: (2408±630)h•ng/ml. Conclusion A simple, rapid, accurate, high sensitivity and repeatability method has been successfully developed, which can analyze the concentration of L-CDL in rat plasma. The method can be used for the investigation of pharmacokinetics of L-CDL in rats.

P-glycoprotein(P-gp) is a multi-drug efflux transporter that plays a significant role in governing the bioavailability of various anti-cancer drugs. P-gp transporter impedes the permeability of drugs through physiological barriers resulting in limited pharmacological response. Modulation of this efflux transporter by various traditional "chemosensitizers" forms a distinctive approach in improving pharmacokinetics and conquering drug resistance. This review summarizes the recent advances in structural information of P-gp interactions with investigated ligands and new strategies to improve the poor efficiency in chemotherapy.

Objective • To optimize the method of the flow cytometry MoFlo Astrios EQ on single-cell sorting in 96-well plate. Methods • Using different aperture nozzles and sorting ways, the 32D, U937, iBMDM and 293T cells were used for single-cell sorting after the precise adjustment of the instrument and various parameters. The hole numbers with single cell and single-cell clones were detected after sorting. Results • In the single-cell sorting mode, the hole numbers with single cell were 83-91 by 70 μm nozzle and 87-93 by 100 μm nozzle. After 7-10 days of culture, the hole numbers with single-cell clones were 36-58 by 70 μm nozzle. In 100 μm nozzle, the hole numbers with single-cell clones were 53-78 by electrostatic charge sorting and 69-81 by straight-down sorting, respectively. Conclusion • In single-cell sorting, a better cell viability and higher cloning rate are observed in 100 μm nozzle and straight-down sorting.