Biomedical Research ; Humans ; MicroRNAs ; Neoplasm Invasiveness ; genetics ; Neoplasm Metastasis ; genetics

Objective To establish a rapid method for the isolation of murine peritoneal macrophages. Methods Abdominal lavage was performed with NS and the peritoneal macrophages was purified from the lavage,which was later transferred into high glucose DMEM cul-ture medium. Cell viability was measured via the typan blue staining. Purity was observed via Wright's stain. Results High purity macropha-ges with typical morphology were obtained. Conclusion A simple and realistic method was set up for the isolation of murine peritoneal mac-rophages.

Objective:To study the clinical efficacy of acupuncture for ischemic stroke and its effect on ET-1 levels in IS cases.Method:The 63 cases were randomized into a treatment group (31 cases),receiving acupuncture and routine method,and control group (32 cases),receiving routine method alone.Seven days constitute a course of treatment,a 2-day interval between two courses.The clinical efficacy and ET-1 level in two groups were compared after four weeks.Result:The total effective rate in the treatment group and control group were 96.8% and 75% respectively,with a statistical significance (P＜0.05);the ET-1 level in blood plasma also with a statistical significance after the treatment (P＜0.05);the pre-treatment and post-treatment ET-1 level in the treatment group showed a statistical significance (P＜0.01),whereas the control group didn't.Conclusion:Acupuncture is effective for ischemic stroke and can lower the ET-1 level in ischemic stroke cases.

Objective To explore the changes of intereferon-?(IFN-?)and interleukin-4(IL-4)in peripheral blood of children with mycoplasma pneumoniae(MP) encephalitis at the acute phase.Methods The peripheral blood concentrations of IFN-? and IL-4 were measured by enzyme-linked immunosorbent assay(ELISA) in 24 cases of children with MP encephalitis at the acute stage.The samples from 24 cases of healthy children were control group.Twenty-four children with MP encephalitis were intravenous drip with azithromycin,at the same time,10 cases received hexadecadrol and 15 cases received gamma globulin.Results The serum concentrations of IFN-? and IL-4 in the mycoplasmal encephalitis group were(98.56?12.93) and(45.55?17.58) ng/L,respectively,which were significantly higher than those in control group [(85.35?6.91) and(26.78?9.89) ng/L] respectively(Pa

A rapid and convenient ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) method for analysis of ribavirin and its two main kinds of metabolites, 1,2,4-triazole-3-carboxamide (TCONH2 ) and 1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxylic acid (RTCOOH), in fresh eggs has been developed and validated. The sample was extracted with acetonitrile water (9 ∶ 1, V/ V), added N-hexane to move liposoluble substances, after centrifugation, the upper solvent was cleaned-up by C18 and GCB, then determined by UPLC-MS / MS coupled with Agilent ZORBAX SB-Aq column (100 mm×3. 0 mm, 1. 8 μm). Over the concentration range of 2. 0-200. 0 μg / L (0. 5-200. 0 μg / L and 5. 0-200. 0 μg / L) for ribavirin (TCONH2 and RTCOOH) in fresh egg matrix, the correlation coefficients (R2 ) of the calibration curves were above 0. 99, and the limits of detection (LODs) were 0. 54 μg / L (0. 09 and 1. 54 μg / L). The limits of quantitation (LOQs) were 1. 79, 0. 31 and 5. 13 μg / L, respectively. At three spiked concentration levels of 5. 0, 10. 0 and 50. 0 μg / L, the recoveries of ribavirin and RTCOOH ranged from 96. 1% to 99. 6%and 42. 9% to 58. 3% . And at three spiked concentration levels of 0. 5, 2. 0 and 5. 0 μg / L, the recoveries of TCONH2 ranged from 75. 9% to 106. 7% . The results of the determination in various real samples showed that the method is simple, accurate, rapid and suitable determination of ribavirin and its two main kinds of metabolites in fresh eggs.

Objective To evaluate the effectiveness of treating glioma in combination of two kinds of drugs by comparing the size of tumors and the survival time of different groups in rat glioma after targeted blood‐brain barrier (BBB) disruption by focused ultrasound under MRI‐guide. Methods The stereotaxis instruments and the 10 μl gas‐tight syringes were used to inject gliosarcoma cells into the targeted area of the brain in 40 male Sprague‐Dawley rats. The glioma‐bearing rats models were established. Rats were divided into 4 groups to receive different treatment :(1) no treatment (control, n = 8), (2) IV Avastin (Avastin only, n =10), (3) IV liposomal doxorubicin (DOX only, n =10), (4) IV Avastin and liposomal doxorubicin (Avastin+DOX, n =10). The SonoVue microbubbles and DOX or Avastin were injected into the tail vein respectively on the 12th day after implantation. The tumor size was measured by MRI on immediacy, once a week after targeted BBB disruption by focused ultrasound, and the life span in rat glioma was recorded. Results The average survival time of different groups in rat glioma was as follows :no treatment(17 ± 4)d, Avastin(20 ± 4)d, DOX(25 ± 5)d, DOX+ Avastin(40 ± 5)d. The tumor size after post‐treatment of different groups in rat glioma was as follows :no treatment(5 7.0 ± 4 3.0)mm, Avastin(4 3.0 ± 2 5.0)mm, DOX(4 1.2 ± 3 1.0)mm, DOX + Avastin(2. 20 ± 1. 30)mm. There was significant increased in average survival time and decreased in tumor size after a combination treatment DOX+ Avastin compared with other groups( P < 0 0.1). Conclusions The microbubble blasting by MRI‐guided focused ultrasound enhances the local tissue permeability and promotes the drug delivery of chemotherapy and anti‐angiogenesis locally in glioma‐bearing rats. Especially, the combination of two kinds of drugs has a synergism efficacy that may reduce tumor growth and increase survival time significantly after BBB disruption.

Objective To evaluate the effectiveness of treating glioma in combination with drugs multiply by comparing the size of tumor and the survival time of different groups in rat glioma after targeted blood-brain barrier (BBB ) disruption by MRI-guided focused ultrasound.Methods The stereotaxis instruments and the 10 μl gas-tight syringes were used to inject gliosarcoma cells into the targeted area of the brain in 50 male Sprague-Dawley rats.The glioma-bearing rat model was established.Each rat received either:(1 )no treatment (control;n =8);(2)single liposomal doxorubicin (DOX;n = 10);(3)multiple DOX (n =10);(4)single Avastin (AVS)and DOX (n =10);(5)multiple AVS and DOX (n =10).The SonoVue microbubble ultrasonic contrast agent and DOX or AVS were injected into the tail vein respectively on day 12 after implantation.The tumor size was measured by MRI on pre-treatment,immediacy and once a week of post-treatment after targeted BBB disruption by focused ultrasound,and the life span in rat glioma was recorded.Results The mediam survival of different groups in rat glioma(The range of the life span 13-90 d):no treatment (7 d);single DOX (12 d);multiple DOX (1 5 d);single AVS + DOX (22 d), multiple AVS+ DOX (30 d).There was significant difference of the groups on mediam survival comparison (P < 0.01 ).The tumor growth pattern after post-treatment of different groups in rat glioma except control:single DOX was noticeable fast and multiple AVS+DOX was visibly delayed comparable to other groups,and finally the tumor size of multiple AVS + DOX even became small.Conclusions The microbubble blasting enhances the local tissue permeability and promotes the drug delivery of chemotherapy and anti-angiogenesis locally in glioma-bearing rats by MRI-guided focused ultrasound.Especially,the combination with drugs multiply has a synergism efficacy that may enhance the effectiveness of chemotherapy,reduce tumor growth,and even become small of the tumor size,and increase survival time significantly after BBB disruption.

Objective To measure the expression of activin receptor-like kinases 1(ALK1)in dermal fibroblasts from patients with systemic scleroderma(SSc)and to estimate its role in the production of fibronectin and plasminogen activator inhibitor-1(PAI-1).Methods Dermal fibroblasts were isolated from the lesions of 12 patients with SSc as well as the normal skin of 14 healthy controls,and subjected to a primary culture.The third-passage fibroblasts were used in the next experiment.Western blot and indirect immunofluorescence technique were utilized to quantify the expression of ALK1.A specific siRNA targeting ALK1 was designed,constructed,and transiently transfected into the control dermal fibroblasts,which were then classified into 2 groups to be cultured with or without the presence of transforming growth factor(TGF)-β1 for 72 hours followed by the detection of fibronectin and PAI-1 expression with Western blot.Results As Western blot and direct immunofluorescence technique showed,both control and SSc fibroblasts showed an expression of ALK1 in the cytoplasm and membrane,and the expression intensity of ALK1 in SSc fibroblasts was significantly higher than that in the control fibroblasts(1.97 ± 0.05 vs.1.12 ± 0.03,t =50.96,P ＜ 0.05).The expression of ALK1,fibronectin and PAI-1 was decreased by 90％,58％ and 31％ respectively in specific siRNA-transfected SSc fibroblasts compared with the control siRNA-transfected fibroblasts.TGFβ1 significantly increased the expression of ALK1,fibronectin and PAI-1 in the control siRNA-transfected fibroblasts,but the increase was markedly inhibited by the siRNA-targeting ALK1.Conlusion TGFβ1 can promote the production of fibronectin and PAI-1 via ALK1 in fibroblasts,and ALK1 may be involved in the development of sclerosis in SSc.

Objective:To investigate the effects and mechanism of β-carotene on inflammatory factors (IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells.Methods:Firstly,RAW264.7 cells of being induced by 4 (5 μg/ml)for 24 h were treated with different concentration of β-carotene (20,40,80,160 pmol/L)for 3 h.The cells viability was measured by MTIT,the mRNA relative expression of IL-1 β,IL-6,TNF-cα was detected by fluorescence quantitative PCR,the secretion capacity of IL-1 β,IL-6,TNF-α was detected by ELISA and the protein relative expression of NF-κB p65 protein was measured by Western blot.Secondly,RAW264.7 cells were induced by LPS(5 μg/ml) and different concentration of PDTC(1,5,10 μg/ml)for 24 h,NF-κB p65 protein was measured by Western blot and inflammatory factors were detected by fluorescence quantitative PCR and ELISA.Finally,compared the changes in the relative expression of inflammatory factors and NF-κB p65 protein between LPS+PDTC group and LPS+PDTC + β-carotene group.Results:Compared with the LPS-induced group,β-carotene could increase the cell viability of LPS-induced RAW264.7 cells and inhibied the relative expression of inflammatory factors and NF-κB p65 protein.Inhibited the relative expression of NF-κB p65 protein could reduce the relative expression of inflammatory factors.Compared with the LPS+PDTC group,LPS +PDTC + β-carotene group could inhibit the relative expression of inflammatory factors significantly (P＜0.05).But,there was little difference about the relative expression of NF-κB p65 protein between this two groups.Conclusion:β-carotene inhibits the relative expression of inflammatory factors(IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells through inhibition of NF-κB p65 protein in NF-κB pathway,this pathway isn't unique.