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Broth microdilution method was hired to measure the minimum inhibitory concentrations (MICs) of luteolin against Trueperella pyogenes,Escherichia coli,Salmonella and Streptococcus in order to evaluate the antimicrobial activity of luteolin in vitro.Meanwhile,the bacteria growth curves in medium containing sub-inhibitory concentration of luteolin were measured in this test.The results indicated that Tureperella pyogenes was the most sensitive to luteolin with a MIC of 0.078 g/L than that of these strains;and Salmonella was also sensitive to luteolin (MIC:1.25 g/L).However,The inhibitory effect of luteolin on Escherichia coli and Streptococcus is relatively weak,and shared the same MIC with 2.5 g/L.Luteolin showed inhibitory effects on the growth curves of all the strains in this test at sub-inhibitory concentration,and the inhibitory effects on the growth curves increased with the concentration of luteolin(P＜0.05).
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Objective To study the expression of HIF-2α and VEGF in colorectal cancer and to investigate the relationship between them and clinicopathologic parameter.Methods Immunohistochemistry staining was conducted to detect the expression of HIF-2α and VEGF protein in 67 samples of colorectal tumor tissues and 67 samples of normal adjacent tissue.Results The expression of HIF-2α and VEGF in colorectal cancer tissues was significantly higher than that in adjacent tissues.The expression of HIF-2α and VEGF increased with the clinical stage and lymph node metastasis.The expression of HIF-2α increased with the tumor volume.The expression of HIF-2α and VEGF was not related to the age,sex,tumor location and tumor differentiation of the patients.HIF-2α was positively correlated with VEGF expression.Kaplan-Meier survival analysis showed that HIF-2α expression was associated with survival,that is,the higher expression of HIF-2α the worse of prognosis was obtained.Conclusion HIF-2α is involved in the process of growth,invasion and metastasis of colorectal cancer.This process may be related to the regulation of VEGF expression.
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Objective To investigate the relationship between serum inflammatory cytokines and atherosclerosis in type 2 diabetic patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods One hundred and three patients with type 2 diabetes were divided into OSAHS group (OSAHS group) and type 2 diabetes mellitus group (non-OSAHS group) according to the results of polysomnography(PSG),and 35 healthy subjects were selected as control group.Serum levels of total cholesterol,triglyceride,fasting plasma glucose(FPG),glycosylated hemoglobin(HbA1c),tumor necrosis factor-α(TNF-α),high-sensitivity C-reactive protein (hs-CRP) and carotid intima-media thickness (IMT) were measured,and their relationship with OSAHS were analyzed.Results The waist circumference,BMI,FPG,HbA1c,IMT,TNF-α and CRP in OSAHS group were significantly higher than those in non-OSAHS group (P＜0.05).Logistic regression analysis showed that,IMT was independently associated with TNF-α and hs-CRP.Conclusion Patients of T2DM with OSAHS have poor blood glucose control and higher incidence of atherosclerosis.High levels of TNF-α and hs-CRP may be involved in the formation of atherosclerosis and plaque occurrence and development.
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Objective To construct a prokaryotic expression vector pET-22b+with Middle East respiratory syndrome ( MERS) coronavirus nuclocapsid protein( NP) gene and to express and purify N protein.Methods N gene amplified by PCR was inserted into the prokaryotic expression vector pET-22b+.Recombinant plasmid was confirmed using DNA elec-trophoresis and sequencing.NP was expressed in E.coli BL21(DE3) by IPTG induced and purified by cation exchange chromatography using the AKTA purification system.Results The NP gene sequence was proved to be correct by sequen-cing and the protein was expressed in both soluble and insoluble forms in E.coli BL21 ( DE3 ) after IPTG induction.The purity and concentration of recombinant protein was improved obviously by cation exchange chromatography and enrich-ment.Conclusion Recombiant NP of high purity and concentration is purified and will facilitate NP functional research.
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BACKGROUND:Our previous studies have shown that strontium-doped calcium polyphosphate containing low-dose strontium appears to have a significant effect on angiogenesis-related behaviors of monocultured umbilical vein endothelial cells and osteoblasts. OBJECTIVE:To investigate the effect of strontium-doped calcium polyphosphate on angiogenesis-related behaviors of umbilical vein endothelial cells and osteoblasts co-cultured, including celladhesion, spreading, proliferation, as wel as the protein secretion of vascular endothelial growth factor and basic fibroblast growth factor from co-culture system in vitro. METHODS:Human umbilical vein endothelial cells and osteoblastic cells (MG63) were utilized in this study. cells from passage 3 were used for preparation of the cel-scaffold constructs. After placed in 24-wel plate at a ratio of 2:1, human umbilical vein endothelial cells and MG63 cells were seeded onto strontium-doped calcium polyphosphate, calcium polyphosphate and hydroxyapatite scaffolds and co-cultured for 7 days. The vascular endothelial growth factor and basic fibroblast growth factor protein levels were determined through a double ligand enzyme-linked immunosorbent assay. The colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay was performed to quantify the effect of scaffolds on cellproliferation. RESULTS AND CONCLUSION:Compared with those on calcium polyphosphate and hydroxyapatite scaffolds, cells on strontium-doped calcium polyphosphate scaffolds attached and spread better with a significantly improved cellproliferation. More importantly, the vascular endothelial growth factor and basic fibroblast growth factor expressions were significantly higher in the strontium-doped calcium polyphosphate group than the other two groups (P<0.05), indicating strontium-doped calcium polyphosphate can up-regulate levels of vascular endothelial growth factor and basic fibroblast growth factor proteins.
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Objective To investigate the differential expression profiles of microRNAs in the liver of 60Co γ-ray irradiated mice using microRNA microarray and to explore their main functions by bioinformatic analysis.Methods After SPF C57BL/6J mice expose to 4 Gy-single whole body radiation,total number of peripheral WBC and the fMNPCE were measured at 3 d.The differentially expressed miRNAs in mouse liver were detected with miRNA microarray,miRNA-124 and miR-34a were confirmed by real time RT-PCR assay.Bioinformatic analysis was applied to explore target genes and the main functions of the differential expressed miRNAs.Results Compared with control group,the total number of peripheral WBC decreased( t = 2.87,P ＜ 0.05 ) ,while the fMNPCE in bone marrow increased ( t =-2.91,P ＜0.05) after 4 Gy γ-ray irradiation.miRNA microarray revealed that 17 miRNAs were differentially expressed,in which 9 up-regulated,8 down-regulated.The expression levels of miR-124 and miR-34a were coincident with the result of real time RT-PCR.GO analysis showed that some pathways including adherens junction and cell cycle were suppressed,while some immune-related pathways were activated.Conclusions miR-34a and miR-194 were involved in the regulation of acute radiation damage,some other miRNAs including miR-124、miR-382 and miR-92a* also played important roles in radiation process.
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We have cloned full-length MDR1 cDNA from human normal liver tissue in previous study. Using this MDR1 cDNA as probe, we observed the MDR1 gene expression in human hepatocellular carcinoma treated with and without chemotherapy by Northern blot. The results showed that expression of MDR1 gene in hepatocellular carcinoma tissues was higher than that in their adjacent normal liver tissues; and enhanced MDR1 gene expression was also observed in hepatocellular carcinoma treated with chemotherapic agents. We also explored a method for quantitative analysis of MDR1 gene expression in hepatocellular carcinoma by polymerase chain reaction. This study suggests that overexpression of MDR1 gene may be responsible for the intrinsic and acquired drug resistance of human hepatocellular carcinoma, and PCR is a preferable method for quantitative analysis of MDR1 gene expression in hepatocellular carcinoma.
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Overexpression of multidrug resistance gene, (MDR1) is responsible for the resistance of anticancer agents in cancer cells. In this study, we designed three pairs of DNA primers to clone MDR, cDNA from human normal liver by polymerase chain reaction. The sites of these primers in MDR cDNA are (I)-64-46 (5') and 1680-1698 (3'); (II) 1471-1489 (5') and 2905-2923 (3'); (III)2729-2747 (5') and 3845-3863 (3). The three reactions were underwent after the human normal liver mRNA was reverse transcripted into single strand DNA. The lengths of PCR products are 1762bp, 1452 bp and 1134bp, respectively. The former two products were subcloned in pBluscript SK, respectively and the latter product was subcloned in pGEM7Z (named as pMDR1.7, pMDR1.4 and pMDRl.l, respectively). Cloned genes were certificated as MDR1 cDNA by sequence analysis. Full-length MDR, cDNA was obtained after further subcloning. Full-length MDR1 cDNA we obtained will be a very important tool in molecular diagnosis and gene therapy of anticancer drug resistance.