Objective To explore the expression of DKK-1 and its clinical value in patients with e-sophageal cancer. Methods The levels of serum DKK-1 were measured by sandwich enzyme linked immu-nosorbent assay(ELISA) in 80 patients with esophageal squamous cell cancer and in 35 healthy subjects. The sensitivities of serum DKK-1, CEA and CYFRA21-1 in the patients with esophageal squamous cell cancer were calculated and analyzed to compare their values in diagnosis of esophageal cancer. Results The serum DKK- 1 levels in patients with esophageal squamous cell cancer were significantly higher than those of healthy subjects(P<0.05). According to the highest value of medicine reference of DKK- 1 (14. 54 ng/ml), its sensitivity and specificity were 66. 25% and 82. 86% respectively; There was a significant difference between the sensitivity of DKK-1 in patients with esophageal squamons cell cancer(66. 25%) and that in healthy subjects(17.14%) (P<0.05). The sensitivitis of serum DKK- 1 in patients with esophageal squamous cell cancer were associated with tumor size, TNM stage and lymph node metastasis(P <0.05),but not associated with gender,age and differentiation grade(P>0.05). And the sensitivity of DKK-1 (66. 25%) was significantly higher than that of CEA, CYFRA21-1 (39. 44% ,25. 35%, P <0. 01). Conclusion DKK-1 played a very important role in the growth and metastasis of esophageal cancer and may be a new tumor marker or an important index in the diagno-sis of esophageal cancer.

Objective To investigate effect of Bailing Capsule combined with Xihuangwan in stage III breast cancer patients of ER, MMP-9, tumor markers and estrogen.Methods 76 Patients with stage III breast cancer were selected and divided into control group anf experiment group.Control group were treated by clinical routine method.Experiment group were treated on the base of the control group with Bailing Capsule combined with Xihuangwan.The ER, MMP-9, tumor markers, estrogen, survival time and quality of life were observed and compared.ResuIts Compared with control group, the survival period of the experiment group were longer(P<0.05),the quality of life score were higher(P<0.05),the ER, MMP-9, E2, E1 level and the serum levels of CA15-3, CA125 and CEA were lower(P<0.05).ConcIusion Bailing capsule combined with Xihuang Pill can effectively reduce stage III breast cancer tumor marker levels, decrease the ER, the expression of MMP-9 and estrogen level and prolong survival time, improve the quality of life, has the vital significance to the clinical therapy for stage III breast cancer.

AIM: To investigate the inhibitory effect of arsenic trioxide on human glioma cell line U251 growth, change of gene expression and intracellular calcium content. METHODS: MTT method was used to observe the growth inhibition effect. Cell cycle, positive rate of proliferation cell nuclear antigen (PCNA), apoptosis associated protein Fas and Bcl-2, and intracellular calcium ion (IECa~ 2+ ) levels were measured by flow cytometry in U251 cells treated with different doses of As_2O_3. Apoptosis was detected with annexin V-FITC+PI dual parameter. RESULTS: As_2O_3 inhibited the growth of U251 cells dramatically. There were obvious dosage-effect and time-effect correlations (P0.05). The cell cycle was arrested in G_2M phase. Apoptosis occurred in U251 cells treated with As_2O_3 by annexin V-FITC+PI dual parameter detection. CONCLUSION: As_2O_3 inhibits the growth of U251 cells in vitro dramatically and induces apoptosis. The mechanism is probably associated with the improvement of Fas expression and IECa~ 2+ levels, decrease in PCNA protein expression and cell cycle arrest.

Objective To investigate the effect of S-1 on serum thymidine kinase-1 ( TK1 ) , vascular endothelial growth factor ( VEGF ) and insulin-like growth factor-1 (IGF-1) and tumor biomarkers, estrgen levels and quality of life in patients with advanced breast cancer.Methods 69 cases with advanced breast cancer from January 2014 to May 2015 in the hospital were selected and randomly divided into two groups.34 cases in control group were treated by conventional therapy, and 34 cases in experimental group were treated with S-1.Serum TK1, VEGF and IGF-1, tumor biomarkers, estrogen level and quality of life score were compared pre-and post-treatment.ResuIts Compared with control group, the levels of VEGF, IGF-1 and TK1 were lower (P<0.05), serum CA125, CEA and CA15-3 concentrations were lower (P<0.05),the LH, E2 and E1 levels were lower (P<0.05) and the survival quality score was higher in experimental group ( P<0.05 ) .ConcIusion S-1 has better clinical curative effect in treatment of patients with advanced breast cancer, and effectively reduce serum TK1, VEGF, IGF-1, tumor biomarker levels, improve the quality of survival, which has important significance.

Objective To explore the optimal protocol of lower-extremity contrast-enhanced MRA (CE-MRA) in the evaluation of diabetic foot.Methods Twenty eight healthy volunteers were scanned by CE-MRA in crus twice with parellel imaging factor (PIF) of 3 or 4.The signal-to-noise ratio (SNR),contrast-to-noise ratio (CNR) and image quality of popliteal artery,posterior tibial artery,anterior tibial artery and peroneal artery were compared.Twenty patients with diabetic foot underwent CE-MRA by both of protocol 1 and 2 in leg,crus and foot.Protocol 1 was the traditional Care-bolus protocol and protocol 2 was the optimized K-space center filling delay-time protocol.The difference of two protocols in venous aliasing and in display of femoral artery,popliteal artery,posterior tibial artery,anterior tibial artery,peroneal artery,dorsalis pedis artery,medial plantar artery and lateral plantar artery were compared.The SNR,CNR of two different PIF sequences were compared by paired t test,and the display of artery of crus was compared by Wilcoxon.The display of vessels and venous aliasing of 2 protocols of diabetic foot patients were compared by Wilcoxon.Results In the images of healthy volunteers with PIF of 3,the SNR were 267±84,174±51,147±42;and the CNR were 232 ±83,139±51,108±39 at popliteal artery,posterior tibial artery and peroneal artery.However,in the images with PIF of 4,the SNR were 239±73,157±53,132±35;and CNR were 206±71,124±50,103±33,respectively.Both the SNR and CNR were higher in the former than in the latter(t values were 2.31 to 4.11,P＜0.05).There was no significant difference in the vessel display between the different PIF volunteers (P＞0.05).In the protocol 1 of patients with diabetic foot,the display of popliteal artery,posterior tibial artery,anterior tibial artery,peroneal artery,dorsalis pedis artery,medial plantar artery and lateral plantar artery,the venous aliasing in crus and foot were 3.40±0.82,2.70±0.80,2.50±1.00,2.20±0.77,2.30±0.92,2.15± 1.04,1.45±0.60,2.20± 1.01,2.20± 1.06.And in the protocol 2,they were 3.85±0.37,3.55± 0.69,3.30±0.92,2.90±0.79,3.30±0.92,3.25±0.79,1.95±1.10,3.70±0.47,3.65±0.49,respectively(P＜0.05).All of these parameters of protocol 2 were superior to protocol 1.Conclusion Using a higher PIF properly,setting the personalized K-space center filling delay-time can contribute to improving the image quality of whole lower-extremity MRA in patients with diabetic foot.

Objective To explore the roles of PKCθ(protein kinase Cθ)signaling pathway on the activation,proliferation and TH1/TH2 cytokines production of the γδT lymphocytes stimulated by Mycobacterium tuberculosis antigen(Mtb-Ag) in vitro.Methods Peripheral blood mononuclear cells(PBMC)were pretreated with or without Rottlerin at 5.0 μmol/L,and stimulated with Mtb-Ag and cultured in rhIL-2 containing medium.After different time of culture,activation molecules and cytokines production of γδT cells were measured by flow cytometry.The proliferation proportion and the percentage of each generation of γδT cells were determined by carboxylfluoreseein diacetate succinmidyl ester(CFSE)labeling technique and flow cytometry.Results After 3d of stimulation with Mtb-Ag,the expression rates of CD69 and CD25 of γδT cells were 46.2%and 45.6%,respectively.Pretreatment of 5.0 μmol/L Rottlerin markedly inhibited the both expressions of CD69 and CD25 in γδT cells(P＜0.01).After stimulation and expansion in vitro for 5,10,and 15 d,the percentages of the γδT cell were 9.6%,54.6%and 82.4%,respectively.There was a few γδT cells in propagation on the 5th day of culture,and almost all γδT cell divisions were above 6 generations on the 10th and 15th day of culture.Pretreatment of the Rotflerin significantly suppressed the γδT cell proliferation,but after 10 d culture,there were still a few parts of γδT cells in propagation.Meanwhile,after 7,14,and 21d of culture,upon stimulation with PMA+Ionomycin,the IFN-γ producing-γδT cells were about 80%at all times.But only after 21d culture,IL-4-producing γδT cells was 2.6%.,The percentage of IFN-γ producing γδT cells markedly reduced in Rottlerin group(P＜0.01).IL-4 secretion of the γδT cells was almost completely blocked.Conclusion PKCO signal pathway involves in activation,proliferation and differentiation of the γδT lymphocytes stimulated by Mtb-Ag.

To understand the genetic origin and variation of porcine epidemic diarrhea virus (PEDV) prevailing in Henan province,a total of 22 PEDV positive samples from suckling piglets with severe diarrhea were collected from 16 pig farms and amplified by RT-PCR.ORF3 genes were then cloned into pMD18-T vector and sequenced.The result of sequencing showed that the length of ORF3 genes all were 675 bp which encoded 224 amino acids.The strains in this study had eight amino acid substitutions when compared with CV777 strain.The nucleotide homologies of the 22 strains were 95.9％-100％ and amino acid homologies were 96.4％-100％.By comparing with those of European strain CV777 and vaccine strain truncated CV777,the nucleotide homologies were 97.1％-97.9％ and 94.8％-95.4％,respectively.Based on the phylogenetic relationship of ORF3 genes,the PEDV field strains and PEDV reference strains could be divided into two groups,and all the field strains identified in this study belong to group 2.It suggested that prevalent PEDV strains in this study seem to be closely related to Henan isolates in previous years,domestic strains,Janpanese strains,American strains as well as Korean strains but differ genetically from European strains or vaccine strains used in China.This experiment analyzed the ORF3 gene sequence features in prevailing of Henan region in recent years,which provides a new support of epidemiology study and protein function research of PEDV ORF3 gene.

ObjectiveTo investigate the role of OPN in human breast cancer cell line ( MDA-MB-231) by using small interfering RNA to specifically knockdown OPN expression. MethodsOPN ShRNA expression vector was stably transfected to MDA-MB-231 cell line.The expression of OPN mRNA and protein were analyzed using reverse transcription polymerase chain reaction (RT-PCR)and Western blot,respectively.The growth of MDA-MB-231 cells were observed by MTT.The effect of OPN siRNA on the transplanted tumor growth and tumor hypoxia were assessed in nude mice. ResultsThe expression level of OPN in MDA-MB-231 cells were significantly lower under hypoxia or normoxia(P ＜ 0.05 ).OPN silence with RNAi significantly inhibited the invasion ability and proliferation of MDA-MB-231 cell lines (P ＜ 0.01 ).Inhibition of OPN with RNAi significantly inhibited the growth ability of MDA-MB-231 cells in vivo(P ＜0.05).The tumor hypoxia significantly decreased(P ＜ 0.05). ConclusionsOPN silence with RNAi can effectively inhibit cell proliferation and tumor growth of MDA-MB-231 cells,and decrease the bypoxia level of MDA-MB-231 transplanted tumor in nude mice.

Objective To investigate the dosimetric performance of COMPASS system,a novel 3D quality assurance system for the verification of nasopharyngeal carcinoma volumetric modulated therapy (VMAT) treatment plan.Methods Eight VMAT treatment plans of nasopharyngeal carcinoma patients were performed with MasterPlan,a treatment planning system (TPS),and then these treatment plans were sent to the COMPASS and MOSAIQ system,a coherent control system,respectively.Comparison of the COMPASS reconstructed dose versus TPS dose was conducted by using the dose volume-based indices:dose received by 95％ volume of target ( D95％ ),mean dose ( Dmean ) and γ pass rate,dose to the 1％ of the spinal cord and brain stem volume ( D1％ ),mean dose of leaf and right parotid ( Dmean ),and the volume received 30 Gy for left and right parotid (V30).COMPASS can reconstruct dose with the real measured delivery fluence after detector commissioning.Results The average dose difference for the target volumes was within 1％,the difference for D95 was within 3％ for most treatment plans,and the γ pass rate was higher than 95％ for all target volumes.The average differences for the D1％ values of spinal cord and brain stem were ( 4.3 ± 3.0) ％ and ( 5.9± 2.9 ) ％ respectively,and the average differences for the Dmean values of spinal cord and brain stem were ( 5.3 ± 3.0 ) ％ and ( 8.0 ± 3.5 ) ％ respectively.In general the COMPASS measured doses were all smaller than the TPS calculated doses for these two organs.The average differences of the Dmean values of the left and right parotids were( 6.1± 3.1 ) ％ and ( 4.7 ± 4.4 ) ％ respectively,and the average differences of the V30 values of the left and right parotids were (9.4 ± 7.5 ) ％ and (9.4 ± 9.9)％ respectively.Conclusions An ideal tool for the VMAT verification,the patient anatomy based COMPASS 3D dose verification system can check the dose difference between the real delivery and TPS calculation directly for each individual organ,either target volumes or critical organs.

ObjectiveTo construct a protein interaction network based on high-throughput sequencing data of liver cancer-related non-coding RNAs, to perform a functional enrichment analysis, and to screen out circular RNAs (circRNAs) participating in the development and progression of liver cancer via the mechanism of competitive endogenous RNA (ceRNA). MethodsThe circRNA-miRNA-mRNA network was constructed using gene expression omnibus (GEO) data based on the ceRNA theory. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) analyses were performed to identify circRNAs with potential ceRNA function and explore their functions. ResultsA total of 9 co-expressed circRNAs, 20 co-expressed miRNAs, and 153 co-expressed mRNAs were screened out from the GEO database, and the liver cancer-related circRNA-miRNA-mRNA network was successfully constructed. The GO analysis revealed 90 biological processes, which mainly involved 12 functional clusters including hepatocyte differentiation, phase-change regulation of cell cycle, and negative regulation of transcription factor activity. The KEGG analysis showed that the co-expressed circRNAs were also involved in the p53 and PI3K-Akt signaling pathways. ConclusionThis study provides new insights for circRNAs mediating the development and progression of liver cancer through the mechanism of ceRNA.