Circ-DDX5 inhibits the proliferation and invasion of human breast cancer cell lines by targeting miR-3940 / 基础医学与临床

Objective To investigate the expression of circular-RNA DDX5(circ-DDX5)in breast cancer tissues and its relationship with the clinical stage of breast cancer patients,and to analyze the regulatory mechanism of circ-DDX5 on the proliferation and invasion of human breast cancer cell line.Methods The expression level of circ-DDX5 in breast cancer tissues and its correlation with the clinical stage of breast cancer patients were analyzed by TCGA database.Bioinformatics analysis and dual-luciferase reporter gene experiments verified the targeting rela-tionship between circ-DDX5 and miR-3940.The correlation between circ-DDX5 and miR-3940 expression in breast cancer tissues was analyzed by TCGA database.The expression level of circ-DDX5 in breast cancer SK-BR-3,MDA-MB-231,BT-549,MCF-7,and HCC-1937 cells was detected by RT-qPCR.The circ-DDX5 over-expression plasmid and negative control plasmid were transfected into MDA-MB-231 cells,which were named circ-DDX5 group and NC group,respectively.The proliferation and invasion of MDA-MB-231 cells in the circ-DDX5 group and the NC group were detected by colony formation assay and Transwell assay.The expressions of proliferation pheno-type protein and invasion phenotype protein of MDA-MB-231 cells were detected by Western blot.The expression level of miR-3940 in MDA-MB-231 cells of circ-DDX5 group and NC group was detected by RT-qPCR.Results The expression of circ-DDX5 in breast cancer tissues was lower than that in adjacent tissues(P＜0.01)and the ex-pression level of circ-DDX5 was negatively correlated with the clinical stage of breast cancer patients(P＜0.01).There was a targeting relationship between circ-DDX5 and miR-3940(P＜0.01).The expression of circ-DDX5 and miR-3940 in breast cancer tissue was negatively correlated(P＜0.01).The expression of circ-DDX5 in human breast cancer cell lines was lower than that in immortalized breast epithelial cells MCF-10A(P＜0.05 or P＜0.01).Compared with the NC group,the over-expression of circ-DDX5 could significantly inhibit the proliferation and in-vasion of MDA-MB-231 cells(P＜0.01),as well as the proliferation phenotype proteins(cyclin C,CDK3)and in-vasion phenotype proteins(Snail,vimentin)expression(P＜0.01)and miR-3940 expression(P＜0.01).Conclu-sions The expression of circ-DDX5 in breast cancer tissues and cells is low.circ-DDX5 inhibits the proliferation and invasion of breast cancer MDA-MB-231 cells by targeting the expression of miR-3940.

Correlation between autophagy and polarization of macrophages in atherosclerosis plaque in arteriosclerosis obliterans amputees / 药学学报

This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-β (TGF-β), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.

Immuno-regulatory effects on murine macrophages by low dose naltrexone (LDW) / 中国免疫学杂志

Objective:The effect of naltrexone on function of murine peritoneal macrophage in vitro was studied in order to illuminate its immune activity futher. Methods:Peritoneal macrophages were divided in three groups:RPMI1640 blank control group, LPS positive control group and NTX treated group. Various phenotypic and functional indices were tested by MTS, flow cytometry technology,phagocytosis experiment and ELISA. Results: Compared with RPMI1640 group,at a LDN exhibits paradoxical properties;the expression of CD64 on surface increased while the expression of CD206 decreased in NTX group;the expression of tumour necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)were increased. Conclusion:The results of experiment had proved that LDN could influence macrophage polarzation, regulate the inflamatory mediators production and affect the phagocytosis function of peritoneal macrophage.

Analysis on Volatile Components in Cuscuta chinensis from Different Habitats by HS-SPME-GC-MS / 中国药房

OBJECTIVE:To establish a method for analyzing the volatile components in Cuscuta chinensis,and compare the difference of the volatile components in C. chinensis. METHODS:HS-SPME-GC-MS was adopted:sampling amount was 1.0 g, extracting fibers was 65 μm PDMS/DVB,equilibrium temperature was 120 ℃,equilibrium time was 15 min,extraction time was 30 min,resolution time was 3 min;GC conditions:the column was HP-5MS quartz capillary column,programmed temperature, inlet temperature was 230 ℃,carrier gas was high purity helium,the flow rate was 1.0 ml/min,splitless injection;MS condi-tions:ion source was electron ionization,temperature was 230 ℃,quadrupole temperature was 150 ℃,electron energy was 70 eV,photomultiplier tube voltage was 1.2 kV,the interface temperature was 280 ℃,and scanning range was m/z 35-550. Com-bined with the qualitative analysis for volatile components of C. chinensis from different habitats by HP ChemStation,the relative content was calculated by peak area normalization,and the data was analyzed by principal component analysis and cluster analysis. RESULTS:Totally 52 components were identified,9 of which were the common components in C. chinensis,namely leaf alcohol, 1-octene-3-ol,3-octanol,malt alcohol,diethyl phthalate,caryophyllene,nonaldehyde,octanol and palmitic acid. sample 1,2,3 were clustered into a group,then clustered with 4,5,6 into a group,sample 7,8,9 was clustered into a group,then clustered with 10,11,12 into a group,and sample 13,14,15 clustered into a group individually. CONCLUSIONS:The method is stable and reliable,and suitable for the rapid analysis of volatile components in C. chinensis;and differences of volatile components in C. chinensis from diflerent habitats are discernible.

Antioxidant Activity of Total Flavonoids of Cinnamomum Septentrionale Hand.-Mzt / 中国药师

Objective:To investigate the antioxidant activity of the total flavonoids of Cinnamomum septentrionale Hand.-Mzt. Methods:A ferricyanide reduction method was used to study the reduction ability. The scavenging activity of the total flavonoids on DPPH· free radical( DPPH·),hydroxyl radicals( ·OH)and superoxide anion free radical( O2-·)was investigated as well. Re-sults:Cinnamomum septentrionale Hand.-Mzt. had good reduction and scavenging activity on the free radicals in a concentration-de-pendant manner. When the concentration of total flavonoids of Cinnamomum septentrionale Hand.-Mzt reached 0. 15 mg·ml-1 ,86%of DPPH free radical( DPPH·),68% of hydroxyl radicals( ·OH)and 59% of ultra oxygen anion free radical( O2-·)were scav-enged. Conclusion:Cinnamomum septentrionale Hand.-Mzt. has good antioxidant activity,and as a natural antioxidant,it is worthy of further research and development.

GC-MS Comparative Analysis of Volatile Compounds Extracted from Rhodiola Crenulata Respectively by HS-SPME and SD / 中国药师

Objective: To compare the volatile compounds extracted from Rhodiola renulata respectively by HS-SPME and SD. Methods:The volatile constituents from Rhodiola crenulata were extracted respectively by HS-SPME and SD, and then the contents and the names were confirmed by GC-MS. Results:Totally 39 compounds were identified from Rhodiola crenulata by HS-SPME while 16 ones were identified by SD. Among them, 4 common compounds were detected. Conclusion: There are some differences between the two methods. Compared with SD, HS-SPME is obviously better because more volatile constituents can be extracted from the herb, furthermore, HS-SPME has notable advantages of higher retrieval matching and sensitivity.