MiR-30e-5p overexpression promotes proliferation and migration of colorectal cancer cells by activating the CXCL12 axis via downregulating PTEN / 南方医科大学学报

OBJECTIVE@#To investigate the regulatory effects of miR-30e-5p on biological behaviors of colorectal cancer cells and the role of PTEN/CXCL12 axis in mediating these effects.@*METHODS@#Bioinformatic analysis was performed to explore the differential expression of miR-30e-5p between colorectal cancer tissues and normal tissues. RT-qPCR was used to detect the differential expression of miR-30e-5p in intestinal epithelial cells and colorectal cancer cells. Bioinformatics and dual luciferase assay were used to predict and validate the targeting relationship between miR-30e-5p and PTEN. Human and murine colorectal cancer cell lines were transfected with miR-30e-5p mimics, miR-30e-5p inhibitor, miR-30e-5p mimics+LV-PTEN, or miR-30e-5p inhibitor + si-PTEN. The changes in biological behaviors of the cells were detected using plate clone formation assay, CCK-8 assay, flow cytometry, scratch healing and Transwell assays. PTEN and CXCL12 expressions in the cancer cells were detected by Western blotting. The effects of miR-30e-5p inhibitor on colorectal carcinogenesis and development were observed in nude mice.@*RESULTS@#Bioinformatic analysis showed that miR-30e-5p expression was significantly elevated in colorectal cancer tissues compared with the adjacent tissue (P < 0.01). Higher miR-30e-5p expression was detected in colorectal cancer cell lines than in intestinal epithelial cells (P < 0.01). Dual luciferase assay confirmed the targeting relationship between miR-30e-5p and PTEN (P < 0.05). Transfection with miR-30e-5p mimics significantly enhanced proliferation and metastasis and inhibited apoptosis of the colorectal cancer cells (P < 0.05), and co-transfection with LV-PTEN obviously reversed these changes (P < 0.05). MiR-30e-5p mimics significantly inhibited PTEN expression and enhanced CXCL12 expression in the cancer cells (P < 0.01), and miR-30e-5p inhibitor produced the opposite effect. Transfection with miR-30e-5p inhibitor caused cell cycle arrest in the cancer cells, which was reversed by co-transfection with si-PTEN (P < 0.05). In the in vivo experiments, the colorectal cancer cells transfected with miR-30e-5p inhibitor showed significantly lowered tumorigenesis.@*CONCLUSION@#Overexpression of miR-30e-5p promotes the malignant behaviors of colorectal cancer cells by downregulating PTEN to activate the CXCL12 axis.

Comparative study of white matter diffusion properties in vulnerable and resistant individuals to continuous attention after short term sleep deprivation / 中华行为医学与脑科学杂志

Objective:To investigate the differences of white matter diffusion properties between vulnerable and resistant individuals to continuous attention after sleep deprivation.Methods:According to the psychomotor vigilance test performance before and after sleep deprivation, the participants were divided into the vulnerable group( n=24) and resistant group( n=25). All participants underwent diffusion tensor imaging (DTI) scans.Tract based spatial statistics(TBSS) was used to compare fractional anisotropy(FA), mean diffusivity(MD), axial diffusivity(AD), radial diffusivity(RD) maps between the two groups.Spearman correlation analysis was conducted by SPSS 24.0 to investigate the relationships between the altered DTI metrics and PVT task performance. Results:(1) Compared with resistant group, FA value of vulnerable group decreased in the body of corpus callosum(x, y, z=-8, 9, 25, t=-7.855), right superior longitudinal fasciculus(x, y, z=-39, -7, 26, t=-6.252), bilateral anterior limb of internal capsule(x, y, z=-13, 8, 13, t=-5.235; x, y, z=12, 8, 3, t=-5.024) and right posterior thalamic radiation(x, y, z=-26, -56, 17, t=-5.469)(TFCE corrected, P<0.05, cluster size≥50 voxel). (2) Compared with resistant group, MD value of vulnerable group increased in the body of corpus callosum(x, y, z=-3, -6, 26, t=7.613), right superior longitudinal fasciculus(x, y, z=-31, -19, 38, t=5.314), bilateral anterior limb of internal capsule(x, y, z=-16, 7, 8, t=6.898; x, y, z=15, 5, 7, t=6.652), splenium of corpus callosum(x, y, z=27, -53, 17, t=6.541), and AD value increased in the right superior longitudinal fasciculus(x, y, z=-33, -19, 39, t=4.892), splenium of corpus callosum(x, y, z=-22, -49, 21, t=5.450), genu of corpus callosum(x, y, z=4, 26, 0, t=4.332), as well as RD value increased in the right superior corona radiata(x, y, z=-17, 1, 33, t=7.558), body of corpus callosum(x, y, z=4, -8, 26, t=6.699), right anterior limb of internal capsule(x, y, z=-12, 7, 3, t=5.212) (TFCE corrected, P<0.05, cluster size≥50 voxel). (3) Correlational analysis revealed that the negative correlations were found between PVT task performance and the FA value in the right superior longitudinal fasciculus( r=-0.492, P<0.001), right anterior limb of internal capsule( r=-0.510, P<0.001), right posterior thalamic radiation( r=-0.502, P<0.001) and body of corpus callosum( r=-0.464, P<0.001). The positive correlations were found between PVT task performance and the MD value in the body of corpus callosum( r=0.500, P<0.001), right superior longitudinal fasciculus( r=0.499, P<0.001), splenium of corpus callosum( r=0.462, P<0.001), right anterior limb of internal capsule( r=0.471, P<0.001), and AD value in right superior longitudinal fasciculus( r=0.643, P<0.001), as well as RD value in right superior corona radiate( r=0.498, P<0.001) (Bonferroni corrected, P<0.003). Conclusion:Differences in the microstructural characteristics of white matter fiber tracts in specific brain regions may constitute the potential neuropathological basis for the phenotypes of vulnerable and resistant individuals to continuous attention after sleep deprivation.

CXC chemokine receptor 4 regulates breast cancer cell cycle through S phase kinase associated protein 2 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of CXC chemokine receptor 4 (CXCR4) on cell cycle of breast cancer and its molecular mechanisms.</p><p><b>METHODS</b>The expression of CXCR4 and S phase kinase associated protein 2 (Skp2) was detected by real-time fluorescence quantitative PCR (fqRT-PCR) and Western blot in breast cancer cells. The expression of signal proteins and the downstream genes of Skp2 was detected by Western blot. The effect of CXCR4, PI3K/Akt pathway inhibitor LY294002 and ERK pathway inhibitor U0126 on cell cycle of breast cancer was detected by propidium iodide staining.</p><p><b>RESULTS</b>Skp2 was significantly down-regulated in CXCR4-downregulated cells and up-regulated in CXCR4-upregulated cells. CXCR4 also regulated the expression of Skp2 and other downstream genes by signaling protein. The proportion of cells in G/Gphase increased and that in S phase declined in CXCR4-downregulated cell, and the effect was more significant when combined with the use of LY294002 or U0126.</p><p><b>CONCLUSIONS</b>CXCR4 can affect cell cycle and inhibit the proliferation of breast cancer cells by regulating Skp2 gene expression through PI3K/Akt and ERK signaling pathway.</p>

Effects of lncRNA RP11-770J1.3 and TMEM25 expression on paclitaxel resistance in human breast cancer cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of long non-coding RNA(lncRNA) RP11-770J1.3 and transmembrane protein 25 (TMEM25) on paclitaxel resistance in human breast cancer MCF-7/PR cell line.</p><p><b>METHODS</b>The expression of lncRNA RP11-770J1.3 and TMEM25 in human breast cancer MCF-7(paclitaxel sensitive) and MCF-7/PR(paclitaxel resistant) cells were detected by quantitative RT-PCR. The synthetic interfering fragments of lncRNA RP11-770J1.3 and TMEM25 were transfected into MCF-7/PR cells. Sulforhodamine B assay was used to detect the sensitivity of MCF-7/PR cells to paclitaxel after interference of lncRNA RP11-770J1.3 and TMEM25. The expression of multidrug-resistance genes and proteins were detected by qRT-PCR and Western blot, respectively.</p><p><b>RESULTS</b>lncRNA RP11-770J1.3 and TMEM25 were highly expressed in MCF-7/PR cells, and were significantly down-regulated after transfection of synthetic interfering fragments. Down-regulation of lncRNA RP11-770J1.3 and TMEM25 enhanced the sensitivity of MCF-7/PR cells to paclitaxel, and inhibited the expression of MRP, BCRP and MDR1/P-gp (all<0.05). Such effects were more significant when lncRNA RP11-770J1.3 and TMEM25 were both down-regulated (all<0.05).</p><p><b>CONCLUSIONS</b>lncRNA RP11-770J1.3 and TMEM25 are highly expressed in MCF-7/PR cells, and the down-regulation of lncRNA RP11-770J1.3 and TMEM25 can enhance paclitaxel sensitivity in MCF-7/PR cells.</p>

Mechanical properties, fluorine release behavior and bacterial inhibition of hydroxyapatite-glass ionomer cement / 实用口腔医学杂志

Objective:To investigate mechanical properties,fluorine release behavior and bacteria inhibition effect of hydroxyapatite-improved glass ionomer cement(HA/GIC).Methods:HA/GIC was prepared with HA mass fraction of 28％.Bending strength and compressive strength of HA/GIC and GIC were measured by an universal material testing machine.Microstructure of the materials was observed by scanning microscope technique (SEM).Accumulation of fluorine release was measured by fluorine ion selective electrode.Bacterial inhibition of total bacteria and Streptococcus mutans was monitored by fluorescence in situ hybridization.Results:Bending strength,compressive strength and fluorine release accumulation of HA/GIC were significantly higher than those of GIC (P ＜ 0.05).There were many.irregular polymerized particle monomers and micro cracks within the GIC.In HA/GIC,hydroxyapatite particles were connected with original composition.Bacterial inhibition of total bacteria and Streptococcus mutans by HA/GIC was more effective than that by GIC.Conclusion:HA/GIC has perfect mechanical properties,fluorine release behavior and bacterial inhibition effect.

Effect of mitochondrial DNA deletion on growth and invasiveness of human lymphoma Namalwa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effect of mitochondrial DNA (mtDNA) deletion on the growth and invasiveness of human lymphoma Namalwa cells.</p><p><b>METHODS</b>ρ(0)-Namalwa cells with mtDNA deletion were generated by treating Namalwa cells with ethidium bromide and confirmed by selective ρ(0) test medium analysis, PCR and Western blotting. The growth of ρ(0)-Namalwa cells was evaluated by MTT assay and cell cycle analysis, and the cell migration and invasiveness were assessed with Transwell assay. Reactive oxygen species (ROS) production and cytosolic Ca(2+) were detected by flow cytometry.</p><p><b>RESULTS</b>ρ(0)-Namalwa cells could grow and divide normally in selective medium supplemented with uridine and pyruvate but not in nonselective medium. PCR did not yield the products of mtDNA, nor was COXII expression detected in ρ(0)-Namalwa cells. ρ(0)-Namalwa cells showed an obvious attenuation of cell proliferation and migration abilities with significantly lowered ROS production and cytosolic Ca(2+).</p><p><b>CONCLUSION</b>The suppressed growth and migration of ρ(0)-Namalwa cells may be the result of decreased ROS production and cytosolic Ca(2+).</p>

Effect of Oxaliplatin on cell cycle of hepatocellular carcinoma cell line HepG2 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of Oxaliplatin (L-OHP) on cell cycle in hepatocellular carcinoma cell line HepG2 and the involved mechanism.</p><p><b>METHODS</b>Inhibitory effect of L-OHP on the proliferation of HepG2 cells was determined by MTT assay. Cell cycle distribution was shown by flow FCM. The expression levels of cyclinD1, CDK2, CDK4, p16, p21, p53 were detected by RT-PCR and Western blot.</p><p><b>RESULTS</b>MTT method revealed that L-OHP inhibited proliferation of hepatocellular carcinoma HepG2 cells in a dose- and time-dependent manner. L-OHP induced S cell cycle arrest in HepG2 cell; down-regulated the levels of CDK4, cyclinD1 and up-regulated the levels of p21, p53. There were no significant changes of CDK2 and p16 after L-OHP treatment.</p><p><b>CONCLUSION</b>L-OHP inhibits the proliferation of HepG2 cells by blocking cell at S stage, which may be resulted from the activity of CDK4, CyclinD1 and p21.</p>

Occlusion of dentinal tubules using tricalcium silicate / 中国组织工程研究

BACKGROUND:A number of in vitro experiments have confirmed that the tricalcium silicate not only can be closely integrated with the dentin through self-curing process, but also can induce dentin remineralization in the physiological environment, thereby effectively blocking the dentinal tubules. OBJECTIVE:To further verify the effects of tricalcium silicate solution on the occlusion of dentinal tubules. METHODS:Thirty-six dentinal discs were made of free first premolars from orthodontic patients, and divided into three pretreatment groups randomly. The teeth were soaked in pretreatment solution for 2 minutes, namely 0.29 mol/L ethylene diamine tetraacetie acid, 6%citric acid, and rinsed ultrasonical y with deionized water 20 minutes, respectively. Every above-mentioned group was randomly assigned into experimental group (tricalcium silicate), control group (sodium fluoride) and blank group, and corresponding materials in each group were used to coat the outer dentinal tubules (2 minutes/time). Then, the dentinal discs were saved in artificial saliva in a 37 observed using scanning electron microscope. Diameter and area of open dentinal tubules were calculated. RESULTS AND CONCLUSION:After pretreatment, the dentinal tubules were at open state;except for the blank control group to maintain the original state, acid etching and ethylene diamine tetraacetie acid pretreatment solutions had a stronger capacity of demineralization, which led to the dentinal tubules open. After the dentinal tubules were treated with sodium fluoride and tricalcium silicate, there were varying degrees of sediments, and open dentinal tubule area and average diameter in the sodium fluoride and tricalcium silicate groups were lower than those in the control group (P<0.05). The dentinal tubule treated with tricalcium silicate was almost entirely closed homogeneously, and occasional y, a single open dentinal tubule was seen. Open dentinal tubule area and average diameter in the tricalcium silicate group were significantly lower than those in the sodium fluoride group (P<0.05). The findings verify that dentin occlusion using tricalcium silicate is superior to that using sodium fluoride;and dentin tubule pretreatment with acid etching or ethylene diamine tetraacetie acid is beneficial to desensitization effects.

Suppression of breast cancer proliferation and induction of apoptosis via AKT and ERK1/2 signal transduction pathways by synthetic polypeptide derived from viral macrophage inflammatory protein II / 华中科技大学学报（医学）（英德文版）

SDF-1α, a ligand for the chemokine receptor CXCR4, is well known for mediating the migration of breast cancer cells. In a previous study we demonstrated that a synthetic 21-mer peptide antagonist of CXCR4 (NT21MP) derived from the viral macrophage inflammatory protein II could antagonize tumor growth in vivo by inhibiting cellular proliferation and inducing apoptosis in breast cancer cells. However, the role of SDF-1α in the signaling pathways underlying the proliferation of human breast cancer cells and associated signaling pathways and inhibiting signal pathways of NT21MP remained unclear. The present study investigated the mechanism of NT21MP on anti-tumor in breast cancer in vitro. The effect of NT21MP on the viability of cells was determined by the MTT assay. Annexin V-FITC and PI staining was performed to detect early stage apoptosis in SKBR3 cells treated with SDF-1α and AMD3100 or NT21MP. Western blotting techniques were used to assay the composition of phosphoproteomics and total proteins present in the SKBR3 breast cancer cells. RT-PCR and Western blotting technique were used to detect the effect of NT21MP and AMD3100 on Bcl-2 and Bax expression. The results indicated that SDF-1α prevented apoptosis and promoted the proliferation of SKBR3 human breast cancer cells. As compared with untreated SKBR3 cells, Treatment with SDF-1α significantly increased cell viability, and NT21MP abolished the protective effects of SDF-1α dose-dependently (P<0.05). There was a significant decrease in the percentage of apoptotic cells after SDF-1α treatment as compared with control group (2.7%±0.2% vs. 5.7%±0.4%, P<0.05). But pretreatment of SKBR3 cells with NT21MP significantly attenuated the antiapoptotic effects of SDF-1α as compared with SKBR3 cells without NT21MP pretreatment. The proliferative and anti-apoptotic effects of SDF-1α in SKBR3 cells were associated with an increase in AKT and ERK1/2 phosphorylation as well as a decrease in Bax expression and an increase in Bcl-2 expression. These changes in intracellular processes were blocked by NT21MP in a dose-dependent manner(P<0.05). In conclusion, NT21MP efficiently inhibits SDF-1α-induced proliferation and antiapoptosis in SKBR3 cells by reducing the levels of phosphorylated AKT and ERK1/2, as well as decreasing the ratio of expression of Bcl-2 relative to Bax.

The expression and significance of TGF-β1 and TGF-β receptor Ⅰ , Ⅱ in experimental rat pulpitis / 中国基层医药

Objective To observe the dynamic expression and location of TGF-β1 and TGF-β receptor Ⅰ ,Ⅱ at different time in an experimental model of rat molars reversible pulpitis, to evaluate the role of TGF-β1 and TGFβ receptor Ⅰ , Ⅱ on pulp injury and repair. Methods The reversible pulpitis animal model was established by drilling the enamel without water cooling and then acid etching dentine. The maxillary and teeth of each animal were made 5μm thick serial slides. The slides were stained with hematoxylin-eosin and SABC immunehistochemical technique and then studied under the light microscope and under image analysis, all the data were statistically analyzed by analysis of variance (ANOVA) and t-test. Results Positive staining of TGF-β1 and TβR Ⅰ ,TβR Ⅱ was found in odontoblasts, pulp fibroblasts and vascular endothelial cells. In pulpitis stage,the expressions of TGF-β1 and TGF-β receptor Ⅰ , Ⅱ were increased markedly, and showed dynamic changes at different time;TGF-β1 show a peak at 5d and followed by decreased significantly. TGF-β receptor Ⅰ ,Ⅱshowed the highest at 3d ,and then decreased slightly. From 0d to 3 d,TGF-β1 and TGF-β receptorⅠ,Ⅱ showed uptrends, but their uptrends had no direct proportion relationship. Conclusion The results presented here suggested that TGF-β1 and TGF-β receptorⅠ,Ⅱ might play an important role in pulp wound and repair. The inflammatory reaction in response to TGF-β1 was mediated by TGF-β receptorⅠ, Ⅱ; Although TGF-β1 showed a possible function for the TGF-β receptor,but a direct proportion relationship couldn't be concluded.

Grape seed extract inhibit proliferation of breast cancer cell MCF-7 and decrease the gene expression of survivin / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of grape seed extract (GSE) on breast cancer cell MCF-7 about the of proliferation and the gene expression of survivin, and to investigate the molecular biological mechanism of inhibition by GSE.</p><p><b>METHOD</b>MTT was used to measure the role of proliferated inhibition of GSE at the dosage of 40, 80, 120, 160, 200 mg x L(-1) with different time. To calculate the IC50 of GSE and to select the suitable concentration and treatment time. The change of cell cycle was detected by flow cytometry. mRNA expression of Survivin was observed by RT-PCR. Luciferase kit was used to observe the change of core promoter of survivin which was inserted in the flourescence report vector. And further to analyse the bind side of transcription factor on the sequence of core promoter of survivin was further analysed by using the microsoft of bioinformatics.</p><p><b>RESULT</b>GSE could inhibit the proliferation of breast cancer cell MCF-7 with time and dosage-dependent (P < 0.05). GSE could arrest the cell cycle in S periods (P < 0.05) and also inhibit the mRNA expression of survivin effectively (P < 0.01); GSE could down-regulate the activity of core promoter of survivin significantly (P < 0.01).</p><p><b>CONCLUSION</b>GSE can inhibit the proliferation of breast cancer cell MCF-7 through arresting the cell cycle in S periods. The mechanism may be that GSE regulate the activity of transcription factor which are related to the activity of core promoter of survivin and decrease the gene expression of survivin.</p>

TrKA-siRNA inhibits the expression of NF-?B and promote apoptosis of breast cancer cell line MCF-7 / 基础医学与临床

Objective Investigate the effect of TrKA variation on the expression of neucleoprotein NF-?B P65 and apoptosis.Methods To construct the expression vector of TrKA small interfering RNA,the recombinant was transfected into MCF-7 cells.the stable cell line expressing TrKA small interfering RNA were selected by G418.The mRNA and protein of TrKA were tested by real-time PCR,Western-blot and Immunohistochemistry.The change of neucleoprotein NF-?B P65 was detected by WB,Flow cytometry was used to observe the cell apoptosis.ResultsThe expression vector of TrKA-siRNA was successfully constructed.The mRNA and protein of TrKA were decreased by 74.7% and 80.5% respectively(P

Observation and Nursing to Hepatic Malignancy Patients Treated by Radio Frequency / 中国实用护理杂志

Treating Hepatic Malignancies with radio- frequency is a new microinvasive method which has been developed in recent years. The patients treated with this method felt less painfull and recover quickly after operation. The author has reported the detailed observaticn to 39 patients with hepatic carcinomas and cooperated with the doctors for patients'psychological nursing in this article. During the treatment,the author tried to explain the aim,method and treating effect of the operation, help the patients to relieve nervousness end worries, observe the patients' inoperative response under cardiopneumograph and guide the patients for the postoperative observation of complications so that the treatment was completed successfully. The author suggests that sentimental support is an important step in the whole course of nursing

Relationship between Occupational Lead Exposure and Spontaneous Abortion / 环境与健康杂志

Objective To analyze and evaluate the relationship between occupational lead exposure and spontaneous abortion.Methods The papers on the relationship of occupational exposure to lead and spontaneous abortion of female workers published in Chinese in 1991-2007 were searched by using computer and manual search,after strict selection,the homogeneity test and integrated analysis for the abstracted data of the eligible studies were conducted by using Rev.Man Statistic Software.The combined RR value was used as the total effect target of each project of meta-analysis.Results In total,14 original researches were included(totally 4 330 subjects and 11 970 controls),compared with the control,the occupational exposure to lead was significantly associated with spontaneous abortion(RR=2.83,95% CI:1.97-4.07)(P

Effects of recombinant viral chemokine vMIP-II on cellular immunity stimulated by LPS / 中国病理生理杂志

AIM: To investigate the produce of intracellular cytokine following short-term in vitro stimulation with vMIP and LPS, and discuss the effect of vMIP to cellular immunity. METHODS: The methods of Cross-linking of radioactivity, ELISA and four-colors flow cytometer were used to test the level of the secretion of chemokine IL-12 and intracellular cytokine IFN-? and IL-4. RESULTS: After treated the PBMCs with vMIP-II, the levels of secretion of IL-12, IFN-? and IL-4 were reduced in the present of LPS by competitively combining chemokine receptor; vMIP promoted CD4+T cell to secrete IL-12, IFN-? and IL-4. CONCLUSION: vMIP-II can protect systemic response of immunity and reduce extremely inflammation by down-regulating proinflammation.

Study of the RNA interference silencing of survivin gene and chemosensitivity to docetaxel in MCF-7 cells / 中国药理学通报

Aim Through interfering the expression of survivin with short hairpin RNA(shRNA) technology,to investigate the effect of downregulation of survivin on cell apoptosis and chemosensitivity to docetaxel in breast cancer MCF-7 cells.Methods(1) Using MCF-7 cells as a model system,three groups were set up transfected with lipofectamine,RNAi control plasmid and survivin RNAi plasmid,respectively.The expression of survivin in MCF-7 cells was measured at transcriptional and translational level by using RT-PCR and Western blot methods.(2) The effect on the cell cycle and apoptosis was analyzed with flow cytometry.(3) The viability of cells applied with different doses of ducetaxel was determined by using the method of 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction(MTT method).Results(1) RT-PCR and Western blot demonstrated that survivin expression was significantly decreased by transfection with RNAi targeting plasmid;the expression proportion was reduced by nearly 75.4% and 79.8%(P