OBJECTIVETo study isolation and culture from SD rat bone marrow mesenchymal stem cells (rBMMSCs) and differentiate into endothelial-like cells (ELCs) with VEGF and bFGF.

METHODSIn vitro rBMMSCs were cultured with the method of percoll (1.073 g/ml) density centrifugation and adherence to plastic dishes. Then they were seeded in LG-DMEM supplemented with 10% fetal bovine serum. The relative biologic characteristics of rBMMSCs including cell morphology, phenotype, growth curve, cell cycle and ultrastructure were studied by inverted microscope, immunocytochemistry, flow cytometry, MTT method, transmission electron microscopy(TEM). The induced cells were identified by immunocytochemistry with CD31, CD34, CD144(VE-cadherin), FITC-UEA-1 and had ability in Dil-ac-LDL uptake and were observed under inverted microscope.

RESULTSThe morphology of rBMMSCs was spindle and has whirlpool. Growth curve of P4 showed that there was difference of latency, activity and flat periods. TEM showed that there were two of rMSCs , the small of both were infantile cells. GO/G1 phase of cell cycle was 95.67% and was suggested that most of cells were not in period of proliferation. The part differentiated cells demonstrated the characters of endothelial-like cells under inverted microscope and showed the expression of CD31, CD144, CD34, and possed the functions of endothelial-like cells staining double-positive for Dil-Ac-LDL and FTIC-UEA-1.

CONCLUSION1.073 g/ml percoll density centrifugation and cultured adherence is an effective approach to obtain rBMMSCs. In vitro, the cells have potential to differentiate into endothelial-like cells

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; Rats ; Vascular Endothelial Growth Factor A ; pharmacology

Objective To observe the distribution and expression changes of cavcelin-1 under shear stress in a turbulence flow channel model mimicking susceptible sites of atheroscleresis in vitro.Methods Human umbilical vein endothelial cells (HUVECs) were inoculated to flow channel and then exposed to distubed shear stress for6,12,24 and 48 hours,respectively.HUVECs inoculated to flow channel without shear stress served as controls.Immunofluorescence staining was performed with SABC-FITC to detect the caveolin-1 distribution and RT-PCR was used to determine the mRNA expression of caveolin-1 gone in these cells.Results Confocal microcopy revealed ellipse-like shaped HUVECs under disturbed shear stress and caveulin-1 didn't move toward to the upstream edge of HUVECs but moved toward to the edge of HUVECs,and expression of caveolin-1 gone in HUVECs was significantly downregulated after exposure to shear stress for 48 hours compared to HUVECs without shear stress stimulation.Conclusion Disturbed shear stress affected disposition and expression of caveolin-1 which might be responsible for shear stress induced endothelial cell pathological atherosclerotic changes.

OBJECTIVE To study the pharmacokinetic and distribution of arctiin in rats. METHODS Each rat was given a single dose at random by oral administration. The arctiin in serum and organs were determined by use of RP-HPLC. All pharmacokinetic parameters were calculated with a 3P87 program. RESULTS After oral administration of arctiin at the dose of 300mg·kg-1, Arctiin plasma C-T curve conform to open two-compartment model. The Pharmacokinetic parameters were as follow: A=(37.374 5±8.964 7)μg·mL-1;B=(6.210 6±1.489 3)μg·mL-1;α=(0.004 3±0.000 9)min-1;β=(0.000 4±0.000 2)min-1;Kα=(0.420 2±0.167 5)min -1;t1/2α=(115.192 6±14.382 4)min ;t1/2β=(1 485.578 1±161.173 3)min;K10 =(0.001 0±0.000 4)min -1;K21=(0.001 4±0.000 6)min -1 ;K12=(0.002 3±0.001 3)min -1 ;Cmax=(41.786 3±7.521 7)μg·mL-1 ;Tmax=(9.891 9±4.341 4)min;AUC=(22 503.272 7±4 120.182 8)μg·min·mL-1. Liver had the highest concentration of arctiin after oral administration. CONCLUSION RP-HPLC method is rapid, sensitive and specific for the research of arctiin pharmacokinetic and its distribution in rats. Arctiin is distributed and eliminated quickly in rats.

OBJECTIVETo observe the effect of 25-hydroxyvitamin D3 on the permeability and ZO-1 expression in normal human airway epithelial cells.

METHODSMTT assay was used to assess the viability of human airway epithelial cell line 16HBE following a 24-hour exposure to different concentrations of 25-hydroxy vitamin D3, and the transepithelial electrical resistance (TER) of the cell monolayer was measured using a Millicell-ERS voltohmmeter. Real-time quantitative RT-PCR was employed to determine the changes of ZO-1 mRNA expression in the cells following the exposures.

RESULTSExposure to 25-hydroxyvitamin D3 resulted in significantly increased permeability of 16HBE cells, but the exspression of ZO-1 showed no obvious changes. 25-hydroxyvitamin D3 at 4×10(-9) mol/L showed the strongest effect in increasing the permeability of cell monolayer.

CONCLUSION25-hydroxyvitamin D3 increases the permeability of normal bronchial airway epithelial cell monolayer in vitro, but this effect is not mediated by upregulation of ZO-1 expression.

Bronchi ; cytology ; metabolism ; Calcifediol ; pharmacokinetics ; pharmacology ; Cell Line ; Cell Membrane Permeability ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Humans ; RNA, Messenger ; genetics ; metabolism ; Zonula Occludens-1 Protein ; genetics ; metabolism

Objective To solve the danger and difficulty in transferring seriously injured victims. Methods The operating principle, construction design, electronic control system and software program flowchart of a robot transfer arm for victim-transfer were introduced.Results and Conclusion The victim didn not have to change their body posture during transfer. The procedure is very simple.A push at only one key is enough,without secondary injury.

AIMTo explore a method to replace the isotope probe in the PCR-Select differential screening Kit with DIG labeled probe.

METHODSDifferential expressed sequences isolated from Suppression Subtractive Hybridization (SSH) was translocated onto nitric fibrin film. Probes were prepared with DIG-11-dUTP mixed in by PCR. Hybridization and coloration followed routine operation procedures of DIG labeled probe. Positive hybridization results were verified with reverse Northern blot.

RESULTSThe positive results from the PCR-Select differential screening Kit improved with DIG labeled probe achieved 90% correspondence verified by reverse Northern blot.

CONCLUSIONThe PCR-Select differential screening Kit improved with DIG labeled probe maintained high specificity while avoiding isotope pollution. It can replace isotope probe completely.

DNA Probes ; Digoxin ; Nucleic Acid Hybridization ; Polymerase Chain Reaction ; instrumentation ; methods ; Reagent Kits, Diagnostic

AIMTo explore the molecular mechanism of heat stress by isolating differentially expressed genes occurred during the course of heat stress and filtrating functional gene segments concerned with heat stress with Suppression Subtractive Hybridization technique.

METHODSThere were 2 groups in the experiment: heat stress group and normal temperature control group. mRNAs were isolated from the liver tissue individually and then transcripted into cDNAs. After cut with endonuclease and ligation with adaptors, SSH was carried out with cDNAs of heat stress group as tester while cDNAs of control group as driver. Products were cloned into T/A vector to form recombined plasmids to transfect competence cells for filtration and appraisal.

RESULTSmRNAs were abundant and of high quality, cDNAs were reverse-transcripted and then cut into segments about 500 bp successfully. Ligation efficiency was satisfied. The subtractive hybridization library was constructed successfully after effective hybridization and 180 segments were primarily isolated from it.

CONCLUSIONConstruction of heat stress differentially expressed genes subtractive library has established the foundation of filtrating heat stress concerned genes, it is of active significance for exploring functional genes that controlled heat stress.

Animals ; DNA, Complementary ; Gene Expression ; Heat Stress Disorders ; genetics ; Liver ; metabolism ; Male ; Nucleic Acid Hybridization ; methods ; Rats ; Rats, Sprague-Dawley ; Transcriptome

Fluoroquinolones are frequently used for the treatment of a variety of infectious diseases, but some adverse effects appeared in the clinical application. By consulting internal and abroad literatures, this review summaried some familiar adverse effects and drug interaction of these drugs, so as to provide reference for rational use of drugs clinically.

OBJECTIVE To study influence of arctiin from seeds of Arctium lappa on hemorheology of experimental rats with the blood stasis syndrone. METHODS The blood hemorheology parameters, Fib, aPTT and PT of experimental rats with the blood stasis syndrone were evaluated using semi-automatic biochemical analysis. RESULTS Arctiin obviously decreased their high shear, middle shear, low shear, the blood viscosity, red blood cell aggregation index, red blood cell rigidity index and reductive viscosity. It also significantly prolonged the time of aPTT and PT and lowed the Fib concentration. CONCLUSION Arctiin apparently ameliorated the blood rheology abnormality and enhanced anti-coagulation effect on experimental rats with the blood stasis.

OBJECTIVETo investigate the clinical indications of asthma control test (ACT).

METHODSA total of 120 asthmatic patients with a diagnosis in line with the American Thoracic Society criteria and treated for over a month were enrolled in this study. The patients were asked to complete a survey to assess their symptoms and asthma attacks, and ACT evaluation was conducted by physicians familiar with ACT evaluation. The patients were classified into two groups based on the pulmonary function test (positive for bronchodilator test and provocation test) or based on disease severity (mild and moderate-to-severe asthma groups). The effect of ACT evaluation was graded as good (no less than 4 item available for evaluation), fair (2-3 items available) and poor (no more than 1 item). To further analyze the ACT sensitivity in relation to different disease severity, 29 asthmatic patients with an initial diagnosis and BDT positivity were included, and the ACT score of the patients with mild, moderate and severe asthma based on FEV1% were compared.

RESULTSIn patients positive for bronchodilator test, good, fair and poor evaluation effects were found in 48, 15, and 5 cases, as compared to 10, 15, and 27 in those positive for provocation test, respectively, showing significant differences between the two groups (P < 0.001). In mild asthma group, good, fair and poor evaluation effects were found in 12, 15, and 18 cases, respectively, significantly different from those in moderate-to- severe asthma group (50, 21, and 4 cases, P < 0.001). ACT scores showed a positive correlation to FEV1% in 29 patients with positive BDT (r = 0.55, P = 0.003). ACT scores had no significant difference between mild and moderate asthma groups (P > 0.05), but showed significant differences between mild and severe groups (P = 0.009) and between moderate and severe groups (P = 0.008).

CONCLUSIONACT is more suitable for evaluating patients positive for bronchodilator test or with moderate to severe asthma.

Adolescent ; Adult ; Aged ; Asthma ; diagnosis ; physiopathology ; Female ; Humans ; Male ; Mass Screening ; Middle Aged ; Predictive Value of Tests ; Respiratory Function Tests ; Sensitivity and Specificity ; Severity of Illness Index ; Surveys and Questionnaires ; Young Adult