Objective To explore the application value of dynamic vascular patterns(DVP) curve and parameter figure in the focal liver lesions by contrast-enhanced ultrasound(CEUS).Methods 83 cases of focal liver lesions diagnosed by pathologically or clinically that had undergone CEUS were analyzed by SonoLiver software.Then DVP parametric images were constructed and the time-intensity curve (TIC) parameters were calculated by TIC and DVP curve.Results Rise time(RT),time to peak(TTP),maximum intensity(IMAX),mean transit (mTT),slope of rise,and the absolute value for slope of half down between malignant and benign lesions groups indicated statistically significant difference (P ＜0.05).94.9％ of type Ⅰ was malignant lesions and 100％ Ⅳ type was benign lesions.The four types of DVP curve(Ⅰ,Ⅱ,Ⅲ,Ⅳ)in malignant and benign lesions were 72.5％,13.7％,13.7％,0 and 6.2％,43.8％,31.2％,18.8％,respectively.The proportion and characteristic of DVP parametric images and DVP curves in malignant and benign lesions groups were consistent.Conclusions DVP curve and parametric images of CEUS can demonstrate the difference of flow perfusion static between lesions area and surrounding liver parenchyma dynamically and directly.These are new qualitative indexes of the differential diagnosis in focal liver lesions.

Objective To investigate the influence of matrix metalloproteinases-9 (MMP-9) on permeability of blood-spinal cord barrier (BSCB) after spinal cord injury (SCI). Methods 68 male C57BL/6 mice were randomly divided into sham group, 2 days group (S2), 7 days group (S7) and 14 days group (S14) after SCI with 17 mice in each group. All the groups received a moderate impacted spinal cord injury except the sham group. Evan's Blue (EB) was administered intraperitoneally to detect the permeability of BSCB. Occludin was analyzed by immunofluorescence, the expressions of occludin and MMP-9 were detected by Western blotting. Results After SCI, BMS score significantly reduced, compared with S2 group, S14 group showed a significant increase (P<0.01). The permeability of BSCB was seriously damaged after SCI. Compared with S2 group, S14 group showed a notable down-regulation in the permeability of injured micro-vessels (P<0.05). The expression of occludin was down-regulated and the expression of MMP-9 was up-regulated 7 days after SCI (P<0.05). Compared with S2 group, S14 group showed a significant up-regulation of occludin and a remarkable down-regulation of MMP-9 (P<0.05). Conclusion After SCI, MMP-9 might mediate the expression of occludin to influence the BSCB permeability.

Objective To observe the effects of melatonin on synaptic plasticity impaired by spinal cord injury in rats. Methods A total of 54 female Sprague-Dawley rats were divided into sham group (n=18), control group (n=18) and melatonin group (n=18). Spinal cord inju-ry model was established with modified Allen's method at T10 (10 g from 25 mm height). The number of neurons and the expression of the Nissl body were detected with immunofluorescence and Nissl staining. The expression of neurofilament-200 (NF-200), brain-derived neuro-trophic factors (BDNF), Synapsin I and growth-associated protein-43 (GAP-43) was detected with Western blotting. Results Seven days af-ter injury, the number of motoneurons, the expression of Nissl body in motoneurons, and the expression of BDNF, Synapsin I and GAP-43 decreased in the control group compared with those in the sham group, and they increased in the melatonin group compared with those in the control group. Conclusion Melatonin can repair the impaired synaptic plasticity, which might promote the functional recovery after spi-nal cord injury.

Objective To investigate the influence of lentivirus mediated short hairpin RNA(shRNA)target to human papilloma virus(HPV)16 E6 on invasive ability of cervical cancer Caski cells.Methods Lentivirus was produced after shRNA target to human papilloma virus(HPV)16 E6 and to nonsense was cloned to lentivirus work vector.Infection ratio was assessed by assay of EGFP positive cells of Caski.Total mRNA of E6 was determined by RT-PCR after Caski cells were infected by lentivirus.The change of E6 protein expression was analyzed by Western blot.The invasive ability of Caski cells was assayed employing Transwell.Results The optimal MOI (Multiplicity of infection)of lentivirus to Caski was 2.5.Total mRNA and protein of E6 were decreased (by 70%and 63%)in interfering group compared with control group.The invasive ability of Caski cells also reduced after infected by lentivirus.Conclusion shRNA mediated by lentivirus can inhibit expression of HPV16 E6 and invasive ability of cervical cancer cells.

China ; Dust ; analysis ; Female ; Humans ; Male ; Manufactured Materials ; Occupational Exposure ; prevention & control ; statistics & numerical data ; Occupational Health

Objective To establish and optimize the technology and method of producing large quantity and high-paclitaxe yield callus of 〖WTBX〗Taxus chinensis var. mairei. Methods Wild maternal tree grown in Lingchuan County of Shanxi Province and cultivated tree grown in Xi′an were used as explant source. And the optimum maternal tree for explant cutting, optimum explant type, basic medium, composition and concentration of growth regulators in medium and so on, which were factors of affecting on callus induction, growth and paclitaxe yield, were examined in a series order. Results The juvenile stem segments were the optimum explants because of their earlier and higher rate callus induction than that of other explants. Medium Y5: MS+2,4-D 4.0 mg/L+KIN 1.0 mg/L or medium B5 Ⅲ: B5+2,4-D 3.0 mg/L+KIN 0.1 mg/L+Phe 0.1 mol/L was confirmed optimum callus induction medium in which callus induction rate had reached to 100%. In callus subculture medium, lower concentration of 2,4-D (0.5—3 mg/L) always increased callus growth, but higher concentration of 2,4-D (8 mg/L) reduced callus growth. When 2,4-D concentration was suitable, callus grown on B5 medium displayed lighter browning and faster tissue growth than that on MS medium. Further more, HPLC analysis confirmed that the paclitaxel yield in callus grown on medium MSⅢ was highest and had reached 0.004% of callus dry weight. In a general condition, the level of paclitaxel in calli derived from juvenile stems of wild maternal tree was higher than that in calli initiated from cultivated maternal tree's juvenile stems. Conclusion The optimization sequence of obtaining a large quantity and high-paclitaxe yield callus of T. chinensis var. mairei are dividing juvenile stem segments from wild maternal tree in May and culturing on medium Y5 or B5 Ⅲ for callus induction. After the calli having been subcultured on the same medium for 8—10 generations, one or two generations are recultured on medium MSⅢ. Finally, the calli with more paclitaxel are obtained by extracting paclitaxel out of it.

BACKGROUND: Apelin/APJ system has a wide range of physiological functions,but its intracellular signal transduction,in particular,apelin receptor desensitization,internalization,resensitization degradation,have still no consistent opinion.OBJECTIVE: To construct eukaryotic expression vector expressing human apelin receptor(APJ)tagged to red fluorescent protein(pDsRED-express-C1),and to determine the expression in human embryo kidney 293 cells.METHODS: The plasmid pcDNA3.1-hAPJ was used as a template for PCR amplification of human APJ.Following PCR amplification the PCR product were removed and enzymatic digestion with EcoR I and BamH I.Same enzymes were used to cut vector pDsRED-express-C1.The digestive product was ligated by conventional methods of connection,then transfected into Competent E.coli TOP10.Single clones were picked plasmid extraction,followed by restriction enzyme digestion and finally DNA sequencing.The recombinant plasmid with correct sequencing was transfected into human embryonic kidney cells,PI staining,followed by the observation under a confocal microscope.RESULTS AND CONCLUSION: PCR amplified a 1.2-kb fragment,which was consistent with the expected size of the human APJ.The pDsRed-hAPJ recombinant plasmid was cut into two fragments,one corresponded to the pDsRED-express-C1 vector size,and the other fragment corresponded to APJ target fragment.Confocal microscopy analysis showed that,APJ was expressed mainly in the membrane of human embryo kidney 293 cells.The pDeRed-hAPJ eukaryotic plasmid expression vector was successfully constructed and effective expression of this fusion protein is achieved,which might be instrumental in the study of displacement and intracellular localization of human APJ.

OBJECTIVETo establish the migraine rheumatism stasis syndrome animal model.

METHODThe rat migraine rheumatism stasis syndrome animal model was established through rheumatism stimulation with manual climate box, 5-HT reduction caused by reserpine and local cerebral vasospasm. General vital signs (activity, weight, eye gum, hair, feeding, excrement), head scratch frequency and image collection were observed to analyze the changes in biological signs of stasis syndrome (tongue image RGB), thrombin and serotonin of model rats.

RESULTThe reserpine group and the reserpine plus rheumatism model group showed significant reduction in blood coagulation time, pain threshold and 5-HT content in blood and brain (P < 0.01); the reserpine plus rheumatism model group showed an increase in eye gum and decreases in activity, feeding, with thin sloppy stool. According to the tough RGB values, the control group showed light red toughs, the reserpine group showed dark purple toughs, the reserpine plus rheumatism model group showed gray toughs, with notable differences in tough RGB values in all three group.

CONCLUSIONThe rheumatism stimulation with manual climate box, 5-HT reduction caused by reserpine and local cerebral vasospasm can be used to induce the migraine rheumatism stasis syndrome animal model, but its modeling assessment method and process shall be further improved.

Animals ; Blood Circulation ; Diagnosis, Differential ; Disease Models, Animal ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Migraine Disorders ; diagnosis ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Rheumatic Diseases ; diagnosis ; physiopathology

A fusion protein of glucagons-like peptide-1 and human serum albumin (GLP-1/HSA) was expressed and secreted into the fermentation broth with recombinant Pichia pastoris. The productivity of expressed GLP-1/HSA could reach 63.6mg/L in 10L fermentor. After concentrated with hollow-fiber ultrafiltration membrane, GLP-1/HSA was purified from fermentation broth by hydrophobic chromatography, negative ion exchange chromatography and gel filtration chromatography in turn. The HPLC analysis showed that the purified GLP-1/HSA had an overall purity of 95.8%. Furthermore, the analysis of in vivo activity indicated that GLP-1/HSA had the bioactivity of native GLP-1, and could significantly reduce blood glucose level 4h after intraperitoneal administration. It was concluded that a great deal of GLP-1/HSA with higher purity could be harvested by Pichia pastoris expression system and the established purification methods. Preliminary studies show a new potential for developing the long-acting GLP-1 analogs for clinical applications.

Two endophytic-bacteria isolates of G18 and F19 were isolated from the stem of Taxus chinensis. The G18 and F19 were respectively classified into Psudomonas sp. and Stenotrophomonas sp. based on biological characteristics and 16S rDNA sequence analysis. The bioactivity analysis showed that the fermented broths of the G18 and F19 exhibited antagonistic activities against three pathogenic bacteria, and had good antagonistic effectiveness to Verticillium dahliae and Colletotrichum gloeosporioides, respectively. The G18 can degrade salicylic acid, and the F19 can do dichlorvos.