Abstract:Objective: To explore the biological function of TIAM1 gene in colorectal cancer cells and its role in cell signaling network by doing bioinformatics analysis. Methods: Firstly, we forecasted the miRNA which interacted with TIAM1 by using miRNA predicting tools ,next analyzed the miRNA which related with the deletion of TIAM1 by using the expression profiles of colorectal cancer cell lines SW480 with deleted TIAM1; then selected the miRNA (hsa-mir-340) which related with TIAM1 from the above two analyses. We forecasted target genes of the miRNA and doing pathway enrichment analysis. We compared the enrichment results and TIAM1 interaction protein pathway enrichment result.We did the pathway enrichment for the gene set whose expression profile logarithmic ratio was greater than 1 compared with that of wild type sw480 cell lines.We did the pathway enrichment using the protein which in protein-protein interaction network of TIAM1.We then did the pathway enrichment for the protein which interacted with TIAM1 directly and indirectly to epithelial mesenchymal transition (EMT) markers named E-cadhrin and Vimentin. Results: All of the direct prediction and indirect prediction show that microRNA hsa-mir-340 can interact with TIAM1 and coordinate with TIAM1 on the function seen from the pathway enrichment result.We analyzed the expression data of the SW480 which deleted the TIAM1 and clarified that the lack of the TIAM1 is related to the over-active of the P53 signal pathway,DNA damage response,Toll-like receptor signal pathway and so on.That means the lack of the TIAM1 may related to the enhance the antitumor effect.According to the pathway analysis results, TIAM1 gene participated in a lot of signaling pathways such as the cell adhesion, cytoskeleton remodeling, cell proliferation and growth, etc. These signaling pathways were possibly related with the phenomenon of the EMT in colorectal cancer. Conclusions: hsa-mir-340 can regulate the expressions of TIAM1.Signal pathway related with TIAM1 has close relationship with cancer metastasis.TIAM1 can influence the function of EMT markers E-cadherin and Vimentin and promote the colorectal cancer occurrence and metastasis by affecting EMT signal pathways such as TGF-β receptor、androgen receptor、NF-kB、MAPK.

Abstract:Objective To observe the expression of interleukin13 (IL13) in Peripheral circulating of polycystic ovary syndrome (PCOS) patients, and analyze the relevance with the body mass index (BMI), reproductive relevant hormone, fasting blood glucose (FBG) and blood fat. Methods Clinical control trial was carried out. All objects of study were assigned to three groups: PCOS with obesity (40 cases), PCOS with non-obesity (35 cases) and normal control (39 cases). The body weight, body height, waist circumference and hip circumference were measured and BMI and waist/hip ratio (WHR) were calculated. ELISA kit for IL-13 was used to test the expression in peripheral circulating. Chemiluminescence’s method was applied to detect FBG, fasting insulin (Ins), cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL) level for the three group objects. Reproductive hormones including follicle-stimulating hormone (FSH), corpus luteum erythropoietin (LH), estradiol (E2), progesterone (P), prolactin (PRL) and testosterone (T) were assayed by Radiation immunoassays (RI). Results The blood level of IL13 in PCOS with obesity was higher than that in PCOS with non-obesity (P = 0.000), and there were significance for both of all compared with that in normal control group (P = 0.000；P = 0.019). Ins, LDL, HDL, TG, TC, IR, E2, FSH and P serum levels were no statistical difference; and WHR, BMI, FBG, LH and T index serum value, both of the PCOS with obesity group and PCOS non-obesity group are higher than those in the control group. There was a moderate correlation for IL13 with BMI(R=0.527, P=0.000) and low-grade correlation with FBG (R=0.449, P=0.000) and T (R=0.452, P=0.000). Conclusion Inflammatory marker IL13 in the peripheral blood PCOS patients increased significantly, and has a positive correlation with BMI, FBG and T indexes.

Abstract:【Abstract】 Objective: To sort the CD133 /CD44 stem cell through MACS from human prostate cancer cell line PC-3 and lay the foundation for the further functional research. Methods: We detected the expressions of CD133 and CD44 on the membrane of PC-3 cell before and after MACS through Flow Cytometry (FCM); then observed the formation of spheres cultured by serum-free medium (SFM) and make subsequent immunofluorescence (IF) assay; distinguished the differences of the morphology、proliferation capacity of PC-3 cell between before and after MACS; detected the expression of prostatic acid phosphatase (PAP) before and after the differentiation assay through immunohistochemical (IHC) and Western blot. Results: The positive expressions of the CD133 and CD44 of the PC-3 cell were (1.33?0.05)% and (0.87?0.06)% through FCM, while they were (84.82?0.07)% and (99.91?0.03)% respectively after MACS; the positive expressions of CD133 and CD44 remained after SFM cultured by immunofluorescence (IF); the proliferation capacity of CD133 /CD44 cell was higher than PC-3 cell (t=11, P=0.008) and CD133 /CD44 cell after induced (t=40.1, P=0.001); the expression of the PAP of the CD133 /CD44 cell after induced by TGF-β was positive, while it was negative in the uninduced CD133 /CD44 cell by IHC and Western blot. Conclusions: The CD133 /CD44 cell had some characteristics of stem cell after MACS from PC-3 cell line through initial functional identification, which could be matting for further explore of the prostate cancer stem cell.

Abstract:Objectives：To develop a method for the preparation of hepatoma glypican 3（GPC3） antibody modified lipid microbubble and to evaluate its efficiency of targeting hepatoma cells in vitro. Methods：Lipid microbubbles were prepared using the mechanical oscil－lation method and the morphology and size of the microbubbles were determined using optical and electron microscopy. The biotin-a－vidin bridge was used to link the GPC3 antibody with the microbubbles and the efficiency of conjugation was determined using flow cytometry. Expressions of GPC3 in 3 kinds of common hepatoma cells were investigated using the immunocytochemical assay. GPC3 antibody modified microbubbles were targeted to hepatoma cells in vitro and the targeting efficiency was determined using confocal analysis. Results：Lipid microbubbles showed a round shape and were evenly distributed without significant aggregation. Diameter of the microbubbles was between 350 nm and 450 nm. 85.05% microbubbles were successfully conjugated with GPC3 antibodies. Fur－thermore，SMMC-7721，HepG2 and Huh7 cells showed positive GPC3 expressions and confocal analysis revealed that the microbub－bles could specifically bind to SMMC-7721 cells. Conclusions：Nanoscale lipid microbubbles conjugated with GPC3 antibody are successfully obtained via biotin-avidin bridging. GPC3 antibody modified microbubbles could selectively target hepatoma cells. Our study provides experimental support for the application of antibody modified nanoscale microbubbles in diagnosis and treatment.

Abstract:Objective：To sort the CD133+/CD44+ stem cell through magnetic bead cell sorting（MACS） from human prostate cancer cell line PC-3 and to lay the foundation for the further functional research. Methods：Expressions of CD133 and CD44 on the membrane of PC-3 cell before and after MACS were detected through flow cytometry（FCM）. Formation of spheres cultured by serum-free medi－um（SFM） was observed and subsequent immunofluorescence（IF） assay was made. Differences in the morphology and proliferation capacity of PC-3 cell before and after MACS were distinguished. Expressions of prostatic acid phosphatase（PAP） before and after the differentiation assay were detected by immunohistochemistry（IHC） and Western blot. Results：Positive expressions of CD133 and CD44 of PC-3 cell were （1.33±0.05）% and （0.87±0.06）% through FCM，while those after MACS were （84.82±0.07）% and （99.91±0.03）% respectively. Positive expressions of CD133 and CD44 remained after SFM detected by IF. Proliferation capacity was higher in CD133+/CD44+ cell than in PC-3 cell（t=11，P=0.008） and CD133+/CD44+ cell after inducement（t=40.1，P=0.001）. Ex－pressions of PAP of CD133+/CD44+ cell were positive after being induced by transforming growth factor-β and were negative in the uninduced CD133+/CD44+ cell based on IHC and Western blot. Conclusions：CD133+/CD44+ cell had some characteristics of stem cell after MACS from PC-3 cell line through initial functional identification，which could be matting for further explore of the prostate cancer stem cell.

Abstract:Objective：To analyze the differential expressions of peripheral serum protein in patients with metastatic and non-metastatic melanoma by proteomic technology and to explore its clinical significance in the treatment of patients with metastatic melanoma. Methods：Totally 101 patients were selected and divided into metastatic melanoma group（n=25） and control group（n=76，31 cases of non-metastatic melanoma and 45 healthy cases）. Seventy-three cases（18 cases of metastatic melanoma，20 cases of non-metastatic melanoma and 35 healthy cases） were randomly extracted as diagnostic model group，the rest 28 cases（7 cases of metastatic melanoma，11 cases of non-metastatic melanoma and 10 healthy cases） were used to verify for model group by blind tests. Matrix assisted laser desorption ionization time of flight mass spectrometry（MALDI-TOF-MS） was applied to test peripheral serum protein. Ciphergen pro－tein chips were used to analyze protein profiling. Results：Mass-to-charge ratios of peripheral serum of patients with metastatic mela-noma against that of control group were 2 598.33，3 646.79，4 605.48，8 603.10，7 756.64 m/z and 9 598.82 m/z. Differences in expres－sions of six protein peaks were statistically significant（all P=0.000）. Diagnostic model of metastatic melanoma was built according to the statics above. Sensitivity of diagnostic model group was 83.33% and specificity was 74.55%. In addition，validation groups were used for this model by blind tests. Sensitivity of metastatic melanoma was 71.43% and specificity was 74.55% based on the model. Conclusions：There are significant changes in expression profile of peripheral serum protein between patients with metastatic melanoma and controls；proteins with relative molecular mass of 1 528.33，2 046.79，3 685.48，4 392.10，5 756.64 m/z and 11 670.82 m/z can be used as biological markers in the treatment of early metastatic melanoma.

Abstract:Objective：To explore the biological function of TIAM1 gene in colorectal cancer cells and its role in cell signaling network by doing bioinformatics analysis. Methods：Firstly，we forecasted the miRNA which interacted with TIAM1 by miRNA predicting tools；then analyzed the miRNA which related with the deletion of TIAM1 by expression profiles of colorectal cancer cell lines SW480 with deleted TIAM1；then selected the miRNA most possibly related with TIAM1 by synthesizing the intersection of the above two analyses. We clarified the signaling pathway participated by these target genes by predicting the target genes of hsa-mir-340 and doing pathway enrichment analysis. We made comparison between enrichment analysis results of target genes and results of proteins related with TIAM1. We clarified the signaling pathway situation which related with the deletion of TIAM1 by selecting the genes in sw480 cell lines with deleted TIAM1，whose expression profile logarithmic ratio was greater than 1 compared with that of wild type sw480 cell lines，and by analyzing the pathway enrichment of these genes. We analyzed the pathway enrichment for the protein in the protein-protein interaction network of TIAM1. We then analyzed the pathway enrichment for the protein which interacted with TIAM1 di－rectly and indirectly to epithelial mesenchymal transition（EMT） markers named E-cadherin and Vimentin. Results：Direct and indi－rect forecast demosnterated that microRNA hsa-mir-340 can interact with TIAM1 and coordinate with TIAM1 on the func－tion. Based on the analysis of SW480 with deleted TIAM1，ex－pressions of P53 signal pathway，DNA damage response，and Toll-like receptor signal pathway were up-regulated，indicating deleted TIAM1 may correlated with tumor inhibitory. According to the pathway analysis results，TIAM1 gene participated in a lot of signaling pathways such as the cell adhesion，cytoskeleton remodeling，cell proliferation and growth，etc and was possibly related with cancer development. TIAM1 and EMT markers E-cadherin and Vimentin regulatory network proteins were the signaling pathway members of transforming growth factor-β（TGF-β） receptor，androgen recep－tor，nuclear factor-?资B（NF-?资B），mitogen-activated protein kinase（MAPK），etc. These signaling pathways were possibly related with the occurrence of the EMT of colorectal cancer. Conclusions：Predicting hsa-mir-340 can regulate the expressions of TIAM1. TIAM1 deletion may correlate with tumor inhibitory and signaling pathway related with TIAM1 has close relationship with cancer metastasis. TIAM1 can influence the function of EMT markers E-cadherin and Vimentin and promote the colorectal cancer occurrence and metastasis by affecting EMT signaling pathways such as TGF-β receptor，androgen receptor，NF-kB，MAPK.

Abstract:Objective：To investigate the effects of STA-21，a specific inhibitor of signal transducers and activators of transcription 3（STAT3） on the proliferation and apoptosis of breast cancer MCF-7B cells. Methods：MTT assay was applied to observe the inhibi－tion of STA-21 on growth of MCF-7B cells after MCF-7B cells being treated with different concentrations of STA-21. Apoptosis and cell cycle of MCF-7B cells were examined by flow cytometry（FCM）. Expressions of P-STAT3，STAT3，CyclinD1 and Bcl-xl protein in MCF-7B cells were detected by Western blot. Results：After cells being exposed to different concentrations（10，30，50 μmol/L）of STA-21，inhibitory rate and apoptotic rate were increased markedly（P=0.000）. It was showed by FCM assay that STA-21 could inhibit the MCF-7B cells at G1 phase. Meanwhile，STA-21 could reduce the expressions of P-STAT3，CyclinD1 and Bcl-xl protein（P=0.000），but STA-21 exerted no effect on STAT3 protein（P=0.367）. Conclusions：Our study demonstrates that STA-21 can inhibit prolifer－ation and promote apoptosis of MCF-7B，mechanism of which may be related with the inhibition of STAT3 activity.

Abstract:Objective：To construct the lentiviral vector expressing shRNA targeting silent information regulator 1（SIRT1） gene and to investigate its effects on proliferation and apoptosis of breast cancer cell MCF-7. Methods：Two shRNA sequences targeting SIRT1 gene were designed and synthesized and were connected to pLentilox3.7-green fluorescent protein（GFP） vector to reconstruct lentivi－ral plasmids pshSIRT1-1 and pshSIRT1-2.Meanwhile，shRNA negative control pshCont targeting no gene was constructed. Recombi－nant lentiviral vector expressing shRNA together with pLP1，pLP2 and pLP/VSVG were co-tranfected into 293FT cells and lentivirus was produced. The titer of virus was tested according to the expression level of GFP in 293FT cells． MCF-7 cells were infected with recombinant lentivirus and the silence efficiency was measured by real-time PCR and Western blot. Proliferation and apoptosis of MCF-7 cells were determined by trypan blue exclusive assay and flow cytometry respectively. Results：Lentiviral vectors containing shRNAs targeting SIRT1 gene were successfully established and corresponding lentiviruses were acquired. Real-time PCR and West－ern blot analysis showed that these two shRNA targeting SIRT1 gene significantly decreased SIRT1 mRNA（P=0.000 2） and protein levels. SIRT1 silencing resulted in marked decrease of proliferation and increase of apoptosis in MCF-7 cells（P=0.006 0）. Conclu－sions：Lentivirus-mediated RNA interference of SIRT1 can significantly inhibit MCF-7 cell proliferation and induce its apoptosis.

Abstract:Objective：To study the effects of phosphoinositide 3kinase（PI3K） inhibitor alone and in combination with epidermal growth factor receptor（EGFR） inhibitor on the proliferation and migration of triple-negative breast cancer（TNBC） cell lines. Meth－ods：Effects of PF-04691502（PF），a PI3K inhibitor，alone or in combination with gefitinib，a EGFR inhibitor，on the proliferation and migration of TNBC cell lines（HCC1937 and MDA-MB 231） were measured by the proliferation，colony formation，wound-healing，flow cytometry and ELISA. Results：PF prevented the proliferation，colony formation and migration of TNBC cell lines，inhibited the protein kinase B（ＰＫＢ or Akt） phosphorylation of these cell lines and retarded the cells in G1 phase（P<0.0５）. Significant synergistic effects were observed when in combination with gefitinib，a EGFR inhibitor. Conclusions：PI3K inhibitor when used alone could pre－vent the proliferation，colony formation and migration of TNBC cell lines and when used in combination with EGFR inhibitors could form significant synergistic effects，mechanisms of which may relate with it’s inhibition of Akt phosphorylation and G1 phase arrest.

Abstract:Objective：To construct Krüppel-like factor 8（KLF8）expression plasmid pcDNA3.1-KLF8 and KLF8 interference plasmid pGPU6/GFP/Neo-KLF8 and to investigate role of KLF8 in regulating angiogenesis related factors in SMMC7721. Methods：KLF8 ex－pression plasmid pcDNA3.1-KLF8 and KLF8 interference plasmid pGPU6/GFP/Neo-KLF8 targeting four interference sites were con－structed and then were screened for the most effective one. Real-time quantitative PCR and Western blot were used to confirm the effects on regulating KLF8 levels. pcDNA3.1-KLF8，pcDNA3.1，pGPU6/GFP/Neo-KLF8 and pGPU6/GFP/Neo-ShNC plasmid were transfected into human SMMC7721 cells separately. mRNA levels of hypoxia-inducible factor（HIF）1-α，vascular endothelial growth factor（VEGF），angiopoietin 1（Ang1），angiopoietin 2（Ang2），angiopoietin receptor Tie2，matrix metallo-proteinase（MMP）2 were investigated by qPCR. Results：Sequencing showed that KLF8 eukaryotic expression plasmids pcDNA3.1-KLF8 was successfully constructed and pcDNA3.1-KLF8 significantly up-regulated KLF8 expressions（relative expression：68.8±6.5）. The most efficient KLF8 interference plasmids pGPU6/GFP/Neo-KLF8 was obtained and pGPU6/GFP/Neo-KLF8 down-regulated KLF8 expressions（relative expression：0.31±0.01）. Compared with those in control group，mRNA levels of VEGF，Ang2，HIF1-α and MMP2 were in－creased（P<0.05） while angiopoietin receptor Tie2 and Ang1 had no significant changes（P >0.05）in experiment group（transfected with pcDNA3.1-KLF8）. Meanwhile，compared with those in control group（transfected with pGPU6/GFP/Neo-ShNC），mRNA levels of Ang2 and HIF1-α were decreased（P<0.05）while VEGF，angiopoietin receptor Tie2，MMP2 and Ang1 displayed no significant changes（P >0.05） in experiment group（transfected with pGPU6/GFP/Neo-KLF8）. Conclusion：KLF8 may regulate angiogenesis in SMMC7721 by regulating several angiogenesis related factors such as VEGF，Ang2，HIF1-α and MMP2.

Abstract:Objective：To observe the inhibitory effects of 125I interstitial brachytherapy combined with cytokine-induced killer（CIK） cells on the growth of human hepatocellular carcinoma（HCC） xenografts in nude mice. Methods：Peripheral blood mononuclear cells were taken from health people and different cytokines were added to promote the mature of CIK cells. CIK cell phenotype was detected by flow cytometry；CD3+CD56+ double positive cells reaching 20% was considered as qualified. BALB/c nude mice were transplanted with SMMC-7721 HCC cell to establish tumor model and the mice were divided into 125I radioactive particle group（group A），CIK cell group（group B），125I radioactive particle + CIK cell group（group C） and blank control group（group D）. Treat－ment was given respectively，diameters of tumors in each group were measured every 4 d and inhibitory effects of different treatments on growth of HCC xenografts were observed. Pathological changes of xenografted tumors were detected by HE staining，histo-pathol－ogy and apoptotic rate of these tumors were examined by TUNEL；expressions of Ki-67 protein were detected by immunohistochemical staining. Results：125I radioactive particle implantation combined with CIK cells intravenously can significantly inhibit the growth of HCC xenografts in nude mice. In group C，tumor volume was （0.42±0.17） cm3，less than those in group A（1.23±0.83） cm3，group B（3.77±1.42） cm3 and group D（6.62 ± 0.53） cm3，with significant differences among groups（F=155.706，P<0.001）.Inhibition rate in group C was 93.71%，significantly higher than those in group A（81.33%） and group B（43.06%）. Immunohistochemical staining re－vealed that mean absorbance value of Ki-67 protein in each group were：group C（0.481±0.063），group A（0.592±0.104），group B（0.669±0.120），group D（0.797±0.113） respectively，with statistically significant differences among groups（F=24.334，P<0.001）. According to the results of TUNEL，apoptotic mean absorbance value in group C was （0.859±0.067），higher than that in group A （0.756±0.055） and group B（0.517±0.051） respectively，with statistically significant differences in each group（F=318.373，P<0.001）. Conclusions：Permanently implanted radioactive 125I particle combined with CIK cells immune therapy can significantly inhibit the growth of HCC xenografts in nude mice，restrain the proliferation of tumor and induce apoptosis in vivo，thus it could become a thera－py for hepatoma.

Abstract:Objective：To explore the influence of newcastle disease virus（NDV） replication in LX-2 cells on the viability and biologi－cal function of hepatic stellate cell（HSC）. Methods：Transforming growth factor-β1（TGF-β1）-stimulated LX-2 cells were infected by different titres of oncolytic NDV-Italien，the proliferation rate of LX-2 cells was detected by MTT assay. Immunofluorescent tech－nique was used to detect the expression of activation markers α-SMA in LX-2 cells. Results：NDV infection in LX-2 cells inhibited the cell proliferation in a dose-dependent manner. The growth inhibition rate of activated LX-2 cells was increased，the expression of activation markers α-SMA of LX-2 cells was down-regulated dramaticlly compared with those of controls. Conclusions：Replication of NDV in activated LX-2 cells inhibits the cell proliferation and attenuates the activation of LX-2 cells.

Abstract:Objective：To evaluate expressions of angiopoietin2（Ang2） protein in human gastric cancer cells SGC-7901 transfected by pEPFP-N1-antisense Ang2 and to investigate whether antisense Ang2 gene could inhibit the vitro growth of human gastric carcinoma cell line. Methods：Two different plasmids including pEGFP-N1 and pEGFP-N1 SGC-7901 antisense Ang2 were separately trans－ferred into an in vitro cultured human gastric cancer cell line SGC-7901 using Lipofectamine2000 transfection technique. After trans－fection，the cells were selected by G418. Then resistant clones were chosen and expanded in DMEM culture medium，with parental SGC-7901 cells as control. RT-PCR and immunohistochemical method were done to determine whether target genes and its proteins had expressed. Cell viability was determined by MTT assay. Apoptosis was determined by flow cytometry with a double dyeing method using FITC-conjugated annexin V and PI. Results：According to results of RT-PCR and immunohistochemical methods，angiopoietin mRNA and its proteins were not expressed in transfected antisense Ang2 gene SGC-7901 cells. MTT assay showed that pEGFP-N1-anti Ang2 transfection group had much lower cell viability than other groups（F=10.39，P=0.002 4）. Meanwhile，pEGFP-N1-anti Ang2 transfection group had an increase in apoptosis rates compared with those of the other groups based on results of flow cytometry（?字2=3 188.98，P＜0.000 1）. Conclusions：pEGFP-N1-anti Ang2 successfully transfects human gastric cancer cells SGC-7901，which in－hibits Ang2 mRNA and protein expressions as well as suppresses the malignant phenotype of human gastric cells.

Abstract:Objective：To explore the efficacy and safety of individualized selection of chemotherapy drug guided by excision repair cross complement group1（ERCC1），ribonucleotide reduc tase 1（RRM1） and tubulin beta 3（TUBB3） in patients with advanced non-small cell lung cancer（NSCLC）. Methods：Totally 47 cases of advanced NSCLC were divided into individualized chemotherapy treat－ment group（group A） and control group（group B） according to histopathological tissue sufficient or not. In group A，mRNA levels of ERCC1，RRM1 and TUBB3 were measured by branched DNA-liquid chip technique and chemotherapy regimen was formulated ac－cording to the measured results. Group B were treated by gemcitabine plus cisplatin. Results：Effective rate of group A was 63.2%，significantly higher than that of group B（P<0.05）. Disease control rates of group A and group B were 78.9% and 50.0% respectively（P<0.05）. Effective rate was significantly higher in patients treated by gemcitabine plus cisplatin in group A than in patients received the same treatment in group B. Quality of life in both groups was improved significantly after treatment（P<0.05） than before treat－ment，but there was no significant difference in improvement rate（P >0.05）. There was no difference in adverse reactions between both groups and all patients were well tolerated. Conclusions：Determination of mRNA levels of ERCC1，RRM1 and TUBB3 by branched DNA-liquid chip technique can guide individualized chemotherapy for advanced NSCLC and significantly improve chemotherapy efficiency； however，further exploration of more effective individualized treatment plan is needed.

Abstract:Objective：To study the expressions of angiopoietin2（Ang2） in gastric carcinoma tissues and its relationship with pathologi－cal stage and to investigate the biological effects of antisense Ang2 cDNA（pEGFP-N1-anti-Ang2 cDNA） on human gastric carcinoma cell line SGC-7901. Methods：Immunohistochemical method was used to detect expressions of Ang2 protein in 53 samples of gastric carcinoma tissues and Western blot was used to detect expressions of Ang2 protein in 10 samples of gastric carcinoma and 10 normal gastric tissues. Two different plasmids including pEGFP-N1 and pEGFP-N1-antisense Ang2 cDNA were transferred seperately into an in vitro cultured human gastric carcinoma cell line SGC-7901 by using Lipofectamine2000 transfection technique. mRNA and protein expressions of Ang2 after transfection in 53 samples of gastric carcinoma tissue were determined by RT-PCR and immunohisto－chemical method. Effects of antisense Ang2 cDNA on cell viability and apoptosis were determined by MTT assay and flow cytometry（FCM）. The three kinds of gastric carcinoma cells were subcutaneously injected into 30 nude mice and were divided into three groups. Velocity of tumor formation and amount of angiogenesis in tumor tissues were detected. Results：Ang2 protein was positively expressed in gastric carcinoma tissues at different pathological stages（F=166.76，P＜0.000 1），while was unexpressed in normal gastric tissues. Based on RT-PCR and immunohistochemical method，SGC-7901 cell transfected antisense Ang2 gene did not express Ang2 mRNA and its protein. MTT assay demonstrated that the group transfected with antisense Ang2 gene had much lower proliferative capacity than other groups（F=10.39，P=0.002 4）. FCM demonstrated that the group transfected with antisense Ang2 gene also had an decreased cell viability than the other groups（ χ2=3 188.980 5，P＜0.000 1）. Tumor formation was slower in cell transfected with antisense Ang2 gene than in other groups（the 6th d：F=10.18，P=0.000 5；the 18th d：F=7.80，P=0.002 1；the 30th d：F=79.58，P＜0.000 1）. Average number of newly formed vessels was fewer in mice transfected with antisense Ang2 gene than in the other groups（the 1th week：F=15.18，P＜0.000 1；the 2nd week，F=3.50，P=0.044 4；the 3rd week，F=39.02，P＜0.000 1）. Conclusions：Ang2 plays an important role in the angiogenesis and tumor formation. mRNA and protein expression of Ang2 in SGC-7901 cells can be inhibited by antisense Ang2 gene. Antisense Ang2 gene can suppress the in vitro growth of human gastric carcinoma cells as well as the growth and angio－genesis in the human SGC-7901 gastric carcinoma.

Abstract:Objective：To explore the anti-tumor effects of dendritic cell（DC） vaccine carrying HPV-16 E6/E7 gene on the CaSki cell transplantable tumor of human cervical cancer in nude mice. Methods：Immature C57BL/6 DC（imDC） was cultured and was infected with recombinant adenovirus vector carrying human papillomavirus-16 E6/E7 gene. Expressions of E7 protein were detected by West－ern blot. CaSki cell transplantable tumor model of human cervical cancer was constructed in nude mice and was treated by cytotoxic T lymphocytes（CTL），acquired from BALB/c mice. Changes in tumor volume were observed. Nude mice were sacrificed and Paraffin-embedded on the 28th d since tumors appeared. Tumor cell morphology was observed in each group by HE staining. Expressions of Bcl-2 and Bax protein of tumor tissues were detected by immunohistochemical techniques. Results：DC was successfully cultured and DC vaccine was prepared. On the 28th d，tumor volume of nude mice was smaller in pAd-E6/E7-DC-CTL-CaSki group than in pAd-mock-DC-CTL-CaSki group，DC-CTL-CaSki group and CaSki group（P<0.05）. Bax protein expressions of tumor tissues detected by immunohistochemical techniques demonstrated that：expressions in pAd-E6/E7-DC-CTL-CaSki group were the highest while in CaS－ki group were the lowest. Bcl-2 protein expressions of tumor tissue detected by immunohistochemical techniques demonstrated that：expressions in pAd-E6/E7-DC-CTL-CaSki group were the lowest while those in CaSki group were the highest. Conclusions：DC vac－cine modified by HPV-16 E6/E7 gene can inhibit the grow of cervical cancer transplantable tumor in nude mice by inducing CTL.

Abstract:Objective：To discuss the prognostic value of excision repair cross complementary gene 1（ERCC1） and breast cancer sus－ceptibility gene 1（BRCA1） in patients with non-small cell lung cancer（NSCLC） who received postoperative platinum-based chemother－apy. Methods：Lung tissues of 80 patients who were eventually diagnosed as NSCLC and were treated by surgery between 2004 and 2008 were collected. Protein expressions of ERCC1 and BRCA1 in lung cancer tissues of 80 NSCLC cases and in adjacent normal lung tissues of 20 cases were detcted by immunohistochemisty. All tissues came from patients with NSCLC who received at least two cycles of platinum-based postoperative adjuvant chemotherapy. Relationships between expressions of two proteins with choice of chemotherapy，disease-free survival（DFS）time and overall survival（OS） time were analyzed. Results：①Positive expression rates of ERCC1 and BRCA1 were significantly higher in lung cancer tissues than in adjuvant normal lung tissues with statistical differences（P=0.036，P=0.041）.Expressions of two proteins were not correlated with patients’ age，sex，clinical stages and pathological classifi－cation. ②Patients in lower ERCC1 expression group had longer DFS time and OS time than those in higher expression group（P=0.004，P=0.032）；similar conclusion was drawn concerning expressions of BRCA1（P=0.023，P=0.006）. ③Patients with lower ERCC1 and BRCA1 expressions had longer DFS time and OS time than those with higher expressions. ④Patients in lower ERCC1 and BRCA1 expression group benefited most from cisplatin-based chemotherapy. Conclusions：ERCC1 and BRCA1 could effectively estimate prognosis of NSCLC patients receiving postoperative adjuvant chemotherapy. Higher ERCC1 and BRCA1 expression may be predictior of patients with NSCLC who have adverse outcomes after postoperative adjuvant chemotherapy.

Abstract:Objective：To investigate significances of variation of telomerase activity and human telomerase reverse transcriptase （hTERT） mRNA in monitoring the inhibition of multiple myeloma（MM） U266 cell proliferation and apoptosis as a result of thalido－mide. Methods：Inhibition of thalidomide on cell proliferation was determined by CCK-8 assay. Percentage of U266 cell apoptosis was measured by flow cytometry. Telomerase activity of U266 cells was performed by TRAP-ELISA. RT-PCR was used to detect the ex－pressions of hTERT mRNA. Results：Thalidomide inhibited the growth of U266 cells and induced cells apoptosis in a time-dose dependent manner. Cells were treated with 10 μg/ml thalidomide for 24 h and 48 h；telomerase activities of U266 cells were respec－tively 7.96±0.09 and 7.65±0.07，showing no significant difference compared with those in control group（P =0.082 0）. Expressions of hTERT mRNA were 2.82±0.03 and 2.31±0.04 respectively，significantly different from those in control group（P=0.033 0）. Telom－erase activities and hTERT mRNA expressions of 50 μg/ml and 100 μg/ml groups were significantly different from those in con－trol group（P=0.002 8）. Conclusions：Thalidomide can inhibit U266 cell proliferation and induce its apoptosis，the mechanism of which is related with telomerase. Change is more sensitive in hTERT mRNA expression than in telomerase activity and it is an ef－fective index when treating MM with thalidomid.

Abstract:Objective：To explore the role of serum intercellular adhesion molecule-1（sICAM-1） in the pathogenesis of hepatitis B virus related liver diseases by detecting the levels of sICAM-1 in patients with hepatitis B virus related liver diseases. Methods：Lev－els of sICAM-1 were measured by enzyme-linked immunosorbent assay（ELISA） in the 120 subjects（30 patients with chronic hep－atitis B，30 patients with liver cirrhosis，30 patients with liver failure and 30 health controls）. Results：Serum levels of sICAM-1 were higher in patients with chronic hepatitis B，cirrhosis and liver failure than in healthy controls（P=0.029，P=0.000 07，P=0.000 1）. In patients with hepatitis B virus related liver diseases，sICAM-1 levels were positively correlated with levels of alanine aminotransferase（ALT），aspartate aminotransferase（AST），total bilirubin（TBIL）（ALT：rs=0.240，P=0.023；AST:rs=0.225，P=0.033；TBIL：rs=0.262，P=0.013）and were negatively correlated with the levels of albumin（ALB），prothrombin activity（PTA）（ALB：rs=-0.297，P=0.005；PTA：rs=-0.218，P=0.039）. Conclusions：sICAM-1 may interfere the procedure of immunological damage of hepatitis B and plays an impor－tant role in the pathogenesis of live fibrosis and cirrhosis. Examination of serum levels of sICAM-1 in patients with hepatitis B virus related liver diseases can be used to judge the patients’ condition.

Abstract:Objective：To observe changes in levels of serum hyaluronic acid（HA），type Ⅲ procollagen amino peptide（PⅢNP），laminin（LN） and collagen Ⅳ（CⅣ） in patients with liver cirrhosis and to explore the relationship between changes of above indexes and de－grees of liver fibrosis. Methods：Blood samples of 204 patients with liver disease were collected in our center from January 2011 to September 2012. The 204 patients（experimental group） included 124 males and 80 females，aged from 26 to 67，48 in average；44 patients with mild chronic hepatitis，50 with moderate chronic hepatitis，31 with severe chronic hepatitis and 79 with posthepatitic cirrhosis. Forty-six health controls from staffs and physical examination persons in our center were enrolled as normal control group，including 25 males and 21 females，aged from 22 to 52，38 in average. Levels of HA，PⅢNP，LN and CⅣ were detected by γ-radia－tion immunity counter. Differences in levels of the indexes between each experimental group and normal control group were analyzed. Results：Levels of HA，PⅢNP，LN and CⅣ were elevated in each experimental group than in normal control group，with statistically significant differences（P<0.01）. Moreover，these indexes were increased gradually followed by the development of disease and the in－crease of HA levels was the most obvious. Conclusions：Serum levels of HA，PⅢNP，LN and CⅣ in patients could be used to indicate the occurrence of liver fibrosis and the degree of fibrosis and could also be used as one of the prognostic indicators after treatment.

Abstract:Objective：To investigate the relationship between plasma copeptin，proadrenomedullin N-terminal 20 peptide （PAMP），N-terminal pro-brain natriuretic peptide（NT-proBNP） levels and severity of chronic heart failure（CHF）. Methods：Seventy-two patients with CHF and 32 non-CHF controls were enrolled. Plasma copeptin and PAMP concentrations were measured by ELISA and concen－tration of NT-proBNP was detected by triaged detection instrument. Cardiac structure and function were evaluated by color Doppler ultrasound. Results：Plasma copeptin and NT-proBNP levels were significantly higher in patients with CHF than in non-CHF controls（P<0.001），which were elevated with the increase of the New York heart association（NYHA） class. Concentration of copeptin was posi－tively correlated with left ventricular end-diastolic diameter（LVEDD） （r=0.508），serum creatinine（r=0.320），NYHA class（r=0.567），NT-proBNP（r=0.412） （all P<0.01） and was negatively correlated with left ventricular ejection fraction（r=-0.663，P<0.01）. There was no significant difference in terms of plasma PAMP levels between patients with CHF and non-CHF controls. Conclusions：Plasma copeptin and NT-proBNP levels are higher in patients with CHF and are closely related with the severity of CHF.

Abstract:Objective：To study the clinical value of serum markers（CA15-3，CA125，CYFRA21-1，CA72-4 and tumor associated ma－terial（TAM）） in the diagnosis of breast cancer at Ⅲ+Ⅳ stages and during postoperative follow-up. Methods：Serum levels of CA15-3，CA125，CYFRA21-1，CA72-4（with electrochemiluminescence immunoassay） and TAM （with biochemical process） were detected in 104 patients with breast cancer（58 patients at stagesⅠ+Ⅱ，46 patients at stages Ⅲ+Ⅳ），60 patients with benign breast tumor and 65 patients with follow-up recurrence. Eighty patients received chemotherapy including those with breast cancer at stages Ⅲ+Ⅳand those with postoperative recurrence（33 patients in invalid group and 47 patients in effective group）. Results：Serum concentration of each marker was obviously higher in breast cancer stages Ⅲ+Ⅳgroup and postoperative recurrence group than in breast benign tumor group and breast cancer stagesⅠ+Ⅱ group. Positive diagnosis rate in breast cancer stagesⅠ+Ⅱ group by single or combined detec－tion was not high. Some commonly used combination detections were compared and analyzed and the combined detection of CYFRA21-1 and CA72-4 was of great significance，which can achieve combined effect of five indexes. CA15-3，CA125，CYFRA21-1，CA72-4 and TAM concentrations were significantly reduced in effective group than in before chemotherapy group and invalid group，with sig－nificant statistical differences（P<0.01）. Conclusions：Combined detection of CYFRA21-1 and CA72-4 is complementary and is great of value in monitoring postoperative relapse and metastasis. Detections of serum CA125，CA15-3，CYFRA21-1，CA72-4 and TAM are of great help for chemotherapy drug efficacy evaluation.

Abstract:Objective：To observe the dynamic changes of plasma high-mobility group box 1（HMGB1） and tumor necrosis factor-α （TNF-α） in patients with severe blunt chest trauma，to assess the severity of injury and to predict its prognostic value. Methods：A total of 57 consecutive blunt chest trauma patients with an abbreviated injury scale≥3 from March 2011 to December 2011 were included and blood samples were taken from patients every other day from d1 to d7 after admission. Plasma concentration of HMGB1 and TNF-α were measured by a quantitative ELISA. Development of post-traumatic complications was observed and their relations with plasma levels of HMGB1 and TNF-α were analyzed. Results：Patients’ median age was （44.5±12.6） years and the median in－jury severity score（ISS） was （18.4±6.02）. Both the plasma levels of HMGB1 and TNF-α had positive correlation with ISS（d1：r=0.29，P=0.068 vs. r=0.507，P=0.016；d3：r=0.578，P<0.001 vs. r=0.411，P=0.021；d5：r=0.645，P<0.001 vs. r=0.365，P=0.07；d7：r=0.683，P<0.001 vs. r=0.321，P=0.019）. There were significantly increases in mean plasma HMGB1 levels on d3，5，7 in patients with ISS≥20 compared with those with ISS<20（all P<0.05）. Plasma levels of HMGB1 showed a higher sensitivity but less specificity than TNF-α in prediction of complications occurrence（P<0.05）. Conclusions：Plasma levels of HMGB1 seem to be most suitable for severity assessment and prediction of post-traumatic complications occurrence in patients with severe blunt chest trauma.

Abstract:Objective：To observe the expressions of interleukin-13（IL-13） in peripheral blood of patients with polycystic ovary syn－drome（PCOS） and to analyze their relationship with the body mass index（BMI），reproductive relevant hormone，fasting blood glucose （FBG） and blood fat. Methods：Clinical control trial was carried out. All patients were assigned to three groups：PCOS with obesity （40 cases），PCOS with non-obesity（35 cases） and controls（39 cases）. Body weight，body height，waist circumference and hip cir－cumference were measured and BMI and waist/hip ratio（WHR） were calculated. Enzyme-linked immuno sorbent assay（ELISA） kit for IL-13 was used to test the expression in peripheral blood. Chemiluminescence method was applied to detect FBG，fasting insulin（Ins），cholesterol（TC），triglycerides（TG），high-density lipoprotein（HDL） and low-density lipoprotein（LDL） level for the patients in three groups. Reproductive hormones including follicle-stimulating hormone（FSH），corpus luteum erythropoietin（LH），estradiol（E2），progesterone（P），prolactin（PRL） and testosterone（T） were assayed by radiation immunoassays（RI）. Results：Expression of IL－13 in peripheral blood was higher in PCOS with obesity than in PCOS with non-obesity（P=0.000） and that was higher in PCOS with obesi－ty and PCOS with non-obesity than in controls（P=0.000；P=0.019）. Serum levels of Ins，LDL，HDL，TG，TC，IR，E2，FSH and P were not statistically different. Serum levels of WHR，BMI，FBG，LH and T were higher in PCOS with obesity and PCOS non-obesity than in controls. Simple correlated regression analysis showed that IL-13 was correlated with BMI（R=0.527，P=0.000），FBG（R=0.449，P=0.000），WHR （R=0.272，P=0.021），LH（R=0.247，P=0.037） and T（R=0.452，P=0.000） and multiple linear regression formula was IL-13=90.814+1.255BMI+0.755WHR+1.696FBG+0.146LH+0.738T. Conclusions：Expression of inflammatory marker IL-13 is in－creased in the peripheral blood of PCOS patients and has positive correlation with BMI，WHR，FBG，LH and T indexes.

Abstract:Objective：To investigate the relationship of plasma protein levels of epidermal fatty acid binding protein（E-FABP） and fatty acid synthase（FASN） with risk factors of human femoral atherosclerosis. Methods：One hundred and seventy-eight patients were detected by color Doppler with intimal-media thickness（IMT） measurement. Then，patients were divided into femoral atherosclerosis group（n=128） and control group（n=50） according to IMT. Fasting protein levels of E-FABP and FASN in plasma were detected and their relationship with femoral atherosclerotic risk factors was measured. Results：Plasma protein levels of E-FABP and FASN in atherosclerotic group were increased compared with those in control group（P<0.05）. Correlation analysis showed that there was a pos－itive correlation between plasma levels of E-FABP and FASN（P<0.05）. E-FABP level was correlated positively with age，body mass index（BMI），triglyceride（TG），adiponection，C reactive protein（CRP） and IMT（P<0.05）. FASN level was correlated positively with age，BMI，fasting plasma glucose，homeostasis model assessment-insulin resistance（HOMA-IR），TG，adiponectin，CRP and IMT（P<0.05）. Plasma levels of E-FABP and FASN were negatively correlated with high-density lipoprotein-cholesterol（P<0.05）. Logistic re－gression analysis showed that age，FASN，E-FABP，HOMA-IR，CRP and TG were the risk factors of femoral atherosclerosis（P<0.05）. Conclusions：Femoral atherosclerosis associated with plasma levels of E-FABP and FASN，which maybe a new circulating biomarker in diagnosis and evaluation of human femoral atherosclerosis.

Abstract:Objective：To study of relationship between postoperative serum thyroid-stimulating hormone（TSH） concentration and re－currence of nodular goiter in Changji Hui autonomous prefecture of Xinjiang and its influencing factors and to further investigate the appropriate level of serum TSH suppression after nodular goiter operation. Methods：Retrospective analysis was made on 781 cases of nodular goiter in People’s Hospital of Changji Hui Autonomous Prefecture. Number of cases within different serum TSH values was counted based on different genders，ages，maximum nodule diameters，nodule numbers and operation modes. Differences in recurrence rates were statistically analyzed. Results：Concentration of serum TSH was significantly higher in those with nodular goiter recurrence than in those without recurrence（3.21±1.34） mIU/L vs. （1.09±0.63） mIU/L（t=20.344 9，P=0.000 0）. With the reduction of postop－erative serum TSH level，recurrence rate in patients with nodular goiter was gradually reduced. But when serum TSH level was reduced to less than 1.0 mIU/L，recurrence rate was no longer declined. When postoperative serum TSH level was lower than 4.5 mIU/L，the larger the thyroidectomy ranges，the lower the recurrence rates. When serum TSH concentration was higher than 4.5 mIU/L，recurrence rate was lower in subtotal thyroidectomy group than in partial thyroidectomy group and lobectomy+isthmusectomies group，with no sig－nificant difference in recurrence rate between the latter two groups（ χ2=0.000，P=1.000）. Conclusions：Excising enough thyroid and suppressing TSH to less than 1.0 mIU/L can maximally prevent the recurrence and minimize the adverse effects due to the application of levothyroxine.

Abstract:Objective：To observe the level of serum leptin in psoriasis and psoriasis with metabolic syndrome（MS） before and after the treatment and to analyze its clinical significance. Methods：Data of height，weight，waistline，psoriasis area and severity index，fasting plasma glucose，blood pressure，blood lipid and MS of psoriasis patients were collected. Patients were divided into two groups according to whether they were suffering from MS. Serum leptin of patients was tested and compared with that of normal group. Results：Duration of psoriasis with MS was longer than that without MS，and patients with MS had poor treatment effect. Before treatment，serum leptin level was higher in all psoriasis patients than in normal controls（P=0.000） and leptin level was the highest in psoriasis with MS. After treatment，leptin level was decreased slightly in psoriasis with MS（P=0.209），but was significantly decreased in psoriasis without MS（P=0.000）. Conclusions：Traditional treatment is ineffective for psoriasis with MS and leptin may involve in the resistance.

Abstract:Objective:To study the expression of Ang-2 in gastric carcinoma tissue, and its relation with pathological stage, To investigate the biological effects of antisense Ang-2 cDNA （pEGFP-N1-anti-Ang-2 cDNA）in human SGC-7901 gastric carcinoma cell line. Methods: Immunohistochemical method was used to detect the expression of Ang-2 protein in 53 samples of gastric carcinoma tissues, and western blot technique was used to the expression of Ang-2 protein in 10 samples of gastric carcinoma tissues and 10 normal gastric tissues. Using the lipofectamine 2000 transfection technique,We transferred separately two different plasmids induding pEGFP-N1,pEGFP-N1- antisense Ang-2 cDNA into an in vitro cultured human gastric carcinoma cell line SGC-7901. The expression of Ang-2 mRNA in 53 gastric carcinoma tissue specimens were determined by RT-PCR. Immunohistochemistry was used to detect the expression of Ang-2 in those tissues as well.The cell viability was detrmined by MTT assay.Apoptosis was determined by flow cytometry . the three kinds of gastric carcinoma cells were subcutaneously injected into 30 nude mice divided into three groups freely. After injection, the tumor mass grew and the angiogenesis of the tumor could be detected. Results:Ang-2 was expressed in gastric carcinoma tissue, while normal gastric tissue was unexpressed. An increased expression of Ang-2 was strong association between the different pathological stages of tumors. On RT-PCR and immunohistochemical method,the SGC-7901 cells transfected antisense Ang-2 gene did not show expression of Ang-2 mRNA and its protein.MTT assay showed the group transfected with antisense Ang-2 gene had a much lower cell viability than other groups(P＜0.05).The group also had an increase in the rate of apoptosis as measured by flow cytometry Compared with other groups(P＜0.01). the tumor grew slower in transfected mice with antisense Ang-2 gene than other groups. The average number of newly formed vessels in the transfected mice with antisense Ang-2 gene was smaller than other groups（P＜0.05）. Conclusions:Ang-2 plays an important role in the angiogenesis and tumor growth, the expression of Ang-2 mRNA and its protein in SGC-7901 cells can be inhibited by antisense Ang-2 gene.The study also show that antisense Ang-2 gene can suppress the in vitro growth of human gastric carcinoma cells. The antisense Ang-2 gene may suppress growth and angiogenesis in the human SGC-7901 gastric carcinoma apparently.