Objective To investigate the differences in tumor volume and metastatic tumors of the liver and regional lymph nodes between contrast-enhanced computed tomography (CT) and diffusion-weighted magnetic resonance imaging (DWMRI) through a comparative analysis,as well the useful information for target volume delineation,and to guide radiotherapy in clinical practice.Methods A total of 40 patients with pancreatic cancer were enrolled and underwent contrast-enhanced CT and DWMRI in the same position.The target volume was delineated,the major axis of the maximum tumor section was measured,and the numbers of liver metastatic tumors and metastatic tumors of the lymph nodes with a diameter of 5-8 mm or＞8 mm were measured based on the CT and DWMRI images.The analysis was performed by using paired t-test or paired Wilcoxon rank sum test.Results The mean gross tumor volume (GTV) delineated by contrast-enhanced CT and DWMRI was 54.95 cm3 and 41.67 cm3(P =0.000),and the mean value-different value was 13.28 cm3.The major axis of the maximum tumor section measured by contrast-enhanced CT and DWMRI were 4.18 cm and 3.94 cm (P=0.000),respectively,and in two patients,dCT was smaller than dDWMRI.A total of 83 liver metastatic tumors were identified by contrast-enhanced CT,and 112 were identified by DWMRI;the liver metastatic tumors detected by contrast-enhanced CT accounted for 74％ of those detected by DWMRI.As for the metastatic tumors of the lymph nodes with a diameter of 5-8 mm or＞8 mm,103 or 46 were detected by contrast-enhanced CT,and 200 or 56 were detected by DWMRI,and the tumors detected contrast-enhanced CT accounted for 52％ or 82％ of those detected by DWMRI.There were significant differences in all data between contrast-enhanced CT and DWMRI.Conclusions GTV and the major axis of the maximum tumor section measured by DWMRI are lower than those measured by contrast-enhanced CT,and contrast-enhanced CT is sensitive in detecting the metastatic tumors of the liver and lymph nodes.However,it is necessary to conduct further controlled experiments with reference to pathology.

Objective To investigate the effect of signal transducers and activators of transcription 3(STAT-3)on sen-sitizing oral squamous cell carcinoma to cis-dichlorodiamineplatinum via downregulating miRNA-21. Methods Tscca and Tca8113P160 human tongue squamous cell carcinoma cell lines were employed in this study. WP1066 was used to suppress STAT-3 signaling pathway. Cells were divided into three groups:dimethyl sulphoxide (DMSO) group, cis-dichlorodiamine-platinum (DDP) group and WP1066+DDP group. Transcription level of miR-21 was assessed by real-time PCR, while the expression levels of STAT-3, p-STAT-3, tissue inhibitor of metalloproteinase-3 (TIMP-3) and matrix metalloproteinase-2/9 (MMP-2/9 ) were evaluated by Western blot assay. Matrigel matrix and transwell assay were used to determine cancer cell colony formation and invasive ability respectively. Expression level of miR-21 was examined by luciferase reporter gene as-say. Results Expression levels of STAT-3, pSTAT-3 and miR-21 were significantly suppressed by WP1066 treatment. The diameters of culture colony in cells treated with WP1066 and DDP were smaller than those in control group. The number of tongue cancer cells that migrated through the transwell membrane in WP1066 and DDP treated group was less than that in control group. Additionally, MMP-2/9 expression decreased while TIMP-3 increased dramatically in both cell lines in WP1066+DPP group compared to the other two groups. Conclusion Reduction of STAT-3 can sensitize oral squamous cell carcinoma to cis-dichlorodiamineplatinum via downregulating miR-21. Our study shows that DDP, in combination with WP1066, might be used as a potential target in the treatment of human oral squamous cell cancer.

Objective: To study the effect of and possible mechanism of knockinng down PI3Kp85α using siRNA in MCF-7 human breast cancer cell line. Methods: Oligofectamine was used to transfect PI3Kp85α siRNA to knock down the PI3Kp85α expression level in MCF-7 human breast cancer cell line in vitro. Real-time PCR was conducted to detect the expression of PI3Kp85α. The effect of PI3Kp85αsiRNA on the growth of MCF-7 cells was measured by MTT. The cell cycle distribution and cell apoptosis were detected by cell flow cytometry. Protein expression was evaluated by immunofluorescence staining and Western blot. Results: The expression of PI3Kp85 α was knocked down with PI3Kp85α siRNA in MCF-7 cells. Cell growth was delayed in PI3Kp85αsiRNA-treated group. Conclusion: The suppressive effect of PI3Kp85αsiRNA on the growth of MCF-7 human breast cancer cell line is significant and PI3Kp85α could be a candidate for gene therapy for breast cancer.

Objective: To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF-7 cells. Methods: The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85 (rAd5-siAKT1-siPI3K), was transfected into human breast carcinoma MCF-7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by real-time PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF-7 cells were measured by MTT, flow cytometry and 2-dementinal and 3-dementional matrigel assay. Results: Recombinant adenovirus vector rAd5-siAKT1-siPI3K dramatically down-regulated AKT1 and PI3K P85 mRNA and protein expressions in MCF-7 cells; the downstream factors PCNA and cyclin D1 were also down-regulated, while P53 was up-regulated. Growth of MCF-7 cells was inhibited by over 50% in rAd5-siAKT1-siPI3K group as measured by MTT assay, and cell cycle was arrested in G_1/G_0 phase compared with untransfected and rAd5-siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5-siAKT1-siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion: shRNA targeting AKT1 and PI3K P85 can significantly down-regulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF-7 cells, and inhibit the growth of MCF-7 cells in vitro.

Tumor immune checkpoint therapy is a clinical treatment strategy developed based on the new principle of the inhibition of negative immune regulation. In this article, the tumor immune checkpoint therapy and the drug delivery strategies were reviewed, mainly including immunity and tumor therapy, tumor immune checkpoint therapy and its mechanism of action, clinical application of tumor immune checkpoint therapy and therapeutic drugs, immune resistance of programmed cell death protein 1 (PD1)/programmed cell death ligand 1 (PDL1) treatment and countermeasures, drug delivery strategies for tumor immune checkpoint therapeutic agents, etc. As a revolutionary new immunotherapy strategy, tumor immune checkpoint therapy has shown obvious superior therapeutic efficacy in a variety types of tumor. However, tumor immune checkpoint therapy is also faced with a big challenge, namely, immunotherapy resistance. With the discovery of new mechanism, the continuous development of new therapeutic drugs and delivery strategies, tumor immune checkpoint therapy is expected to further improve the clinical efficacy of tumor.

Objective To study the mechanism of miR-21 in regulating the invasion of human glioblastoma (GBM) cells in vitro. Methods The transfection reagent oligofectamine was mixed with antisense miRNA-21 (AS-miR-21) and nonsense oligodeoxyribonucleotides (ODN), respectively, and then, they were added into the medium of U251 GBM cell line as AS-miR-21 treatment group and nonsense ODN treatment group, respectively; control group (treated with PBS) was also established.MiR-21 luciferase reporter assay was used to detect the miR-21 knocking down effect. Matrigel cell growth assay and Transwell assay were used to determine the invasion and migration abilities of U251 cells. Western blotting was employed to test the expressions of invasion-related proteins (FAK,MMP-9/2, TIMP-1 and Tubulin-α); immunofluorescence was also employed to observe the morphology of Tubulin-α protein in GBM cells. Results Luciferase intensity in as-miR-21 treated U251 cells was significantly suppressed as compared with that in the control group and nonsense ODN treatment group (P＜0.05). The diameter of cultured clone in as-miR-21 treated U251 cells was smaller than that in the controls and nonsense ODN treatment group (F=102.819, P=0.000). Decreased cells via the transwell member in thc AS-miR-21 treatment group were detected as compared with those in the controls and cnonsensc ODN treatment group (F=243.465, P=0.000). The expressions of FAK, MMP-2/9 weredown-regulated and that of TIMP-1 was up-regulated in the AS-miR-21 treated tumor cells as compared with the other 2 groups (P＜0.05). No obvious changes were noted on the expression of Tubulin α,however, the morphology of Tubulin α protein in the AS-miR-21 treatment group changed. Conclusion High expression of miR-21 induce the ability of U251 GBM cell invasion and miR-21 can be taken as a candidate for gene therapy of human glioma.

OBJECTIVETo study the effects of c9,t11-conjugated linoleic acid on the killing ability of macrophage to B16-MB cells in C57 mice and explore its possible mechanism.

METHODSThe five levels of CLA was designed as 0, 25, 50, 75, 100 micro mol/L. After macrophage was treated with CLA for 24 h, the killing ability of macrophage on B16-MB cells was evaluated by MTT, The expression of C57 mice macrophage cytokine IL-6, TNF-alpha and iNOS mRNA was detected by RT-PCR. The expression of Erk protein was examined by Western Blot assay.

RESULTSThe inhibitory effect of macrophage on tumor cell depend on the treatment of the increased c9,t11-CLA level, at the same time, the expression of IL-6, TNF-alpha and iNOS mRNA increased, the expression of Erk decreased with the elevating dose of CLA.

CONCLUSIONSc9,t11-CLA could increase the killing ability of macrophage in mice to B16-MB cells, and it was associated with induction of IL-6, TNF-alpha and iNOS mRNA expression. We speculate that antitumor ability of CLA may be associated with taking part in body immune regulation action, and the effects of CLA on the killing ability of murine macrophage to B16-MB cells was not associated with the MAPKErk pathway.

Animals ; Blotting, Western ; Cell Division ; Cell Line, Tumor ; Coculture Techniques ; Dose-Response Relationship, Drug ; Interleukin-6 ; genetics ; Linoleic Acids, Conjugated ; pharmacology ; Macrophages ; drug effects ; physiology ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred C57BL ; Mitogen-Activated Protein Kinases ; metabolism ; Nitric Oxide Synthase ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; genetics

Objective To establish a quick,economical and reproducible method for high-quality RNA extraction from pancreas.Methods We utilized TRIzol Reagent and liquid nitrogen to isolate total RNA from the rat pancreas.The RNA quality was determined by detection of its content and optic density(A) at 260/280nm,and electrophoresis in 1% non-denatured agarose gel.Then reverse transcription-polymerase chain reaction(RT-PCR) was performed to detect expression of the pancreas-specific genes.Results The content of the total RNA extracted from the rat pancreas reached 3-6?g/mg pancreatic tissues,and A260/280 ratio was 1.75-1.89.Electrophoresis of the total RNA showed 28S and 18S rRNA bands with clear smear between them.The RT-PCR products of pancreas-specific genes including insulin 1,glucagon,?-amylase and housekeeping gene ?-actin all exhibited clear bands on 1% agarose gel,which were located in the expected positions,respectively.Conclusion These results suggest that we have successfully isolated the high-quality and intact RNA from the rat pancreas with TRIzol Reagent and liquid nitrogen.The extracted total RNA can be used in RT-PCR for pancreatic gene expression.

Objective To investigate the effect of micro RNA-21 (miRNA-21) knocking on the Tb3.1 human tongue squamous cell carcinoma growth. Methods Anti-sense miRNA-21 oligonucleotide was delivered with oligofectamine to suppress Tb 3. 1 tongue cancer cell growth in vitro. Real-time polymerase chain reaction (PCR) was conducted to detect the miRNA-21 expression after transfection. Methyl thiazolyl tetrazolium(MTT) assay was used to determine Tb 3. 1 cell survival rate. Apoptosis were examined by flowcytometry. Matrigel matrix and transwell assay were used to determine Tb 3.1 cell colony formation and migration ability. Antigen KI-67 (Ki67), B cell lymphoma (Bcl-2), phosphatase and tensin homolog (PTEN), matrirx metalloproteinase 2(MMP-2, MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) protein expression in Tb 3. 1 cell were measured by Western blotting. Results miRNA-21 expression was decreased in miRNA-21 antisense oligonucleotide (ASODN) group. The survival rate of Tb 3. 1 cells with AS-miRNA-21 transfection was significantly suppressed (F=27.02, P = 0.00) and early phase apoptosis(F =26. 641 ,P = 0. 001) induced in Tb 3.1 cell. Ki67, Bcl-2, MMP-2 and MMP-9 protein weredown regulated while PTEN and TIMP-1 protein expression was increased. Conclusions Blocking miRNA-21 expression in Tb3.1 cell could suppress cancer cell growth in vitro and miRNA-21 can serve as a novel target candidate for human tongue cancer gene therapy.

It is known that silibinin has antioxidant, anti-tumor, anti-inflammatory and immunomodulatory effects, and which is widely used for liver damage caused by a variety of reasons. In recent years, it is found that silibinin has potential anti-allergic reactions. However, even larger doses of silibinin still show no significant side effects . The rare literature reports that silibinin can cause allergic reactions. The paper reports a middle-aged patient who orally took silibinin for the prevention of tuberculosis chemotherapy-induced liver damage, and he occurred symptoms of lip pain and anabrosis, foreign body sensation, and difficulty eating one day after treatment. The patient was misdiagnosed as"vesicular stomatitis"and was treated by anti-viral therapy. The patient was discharged from the hospital after treating allergic reactions. As a safe and effective drug for prevention of liver damages in clinic, silibinin should be alert to induce possible allergies when there are local skin manifestations such as lip pain and anabrosis.