Objective To explore the effects of COX-2 inhibitor Mavacoxib on proliferation， apoptosis and related signaling pathways of human osteosarcoma stem cells. Methods The MG-63 cells were cultured in vitro to obtain osteosarcoma stem cells. The MTT assay was used to detect the proliferation inhibition rates after treatment with Mavacoxib（0， 1， 10， 50， 100 μmol/L）. Meanwhile， Annexin-FITC/PI double staining and PI staining were used to detect the apoptotic rates at 24， 48 h and cell cycle at 48 h after different concentrations of Mavacoxib. The effects of Mavacoxib on the Wnt/β-catenin and PI3K/Akt signaling pathway were measured by Western blotting. Results In the range of 10-100 μmol/L， Mavacoxib could improve the proliferation inhibition rates of osteosarcoma stem cells in a dose and time dependent manner with statistical significant difference（P＜0.05）. Compared with 0 μmol/L， the apoptotic rates and the percentage of G0/G1 phase cells were increased， the proportions of S phase and G2/M phase were decreased in other concentrations（P＜0.05）. After the treatment of Mavacoxib， the levels of PTEN in the PI3K/Akt pathway were increased， the levels of Akt in the PI3K/Akt pathway and the levels of β-catenin， C-myc and Cyclin D1 in the Wnt/β-catenin pathway were decreased（P＜0.05）.Conclusion Mavacoxib has toxic effect on human osteosarcoma stem cells， and can induce apoptosis and cell cycle arrest and inhibit the activation of Wnt/β-catenin and PI3K/Akt signaling pathway.

Objective To investigate the expression levels of retinoic acid receptor 3 （RARRES3） and migration and invasion enhancement factor 1 （MIEN1） in non-small cell lung cancer （NSCLC） tissues and their relationship with clinical pathological features and prognosis. Methods In this study， 86 cases of NSCLC tissues and 72 cases of adjacent tissues were collected from our hospital. The expression of RARRES3 and MIEN1 in cancer tissues and adjacent tissues were detected by immunohistochemical SP method， and the expression level was divided into low expression and high expression level according to the two-grade score. The relationship between the expression level of two protein and clinical pathological parameters （gender， age， pathological type， tumor size， differentiation type， TNM stage， T stage， smoking history and lymph node metastasis） were analyzed. We further investigated the correlation between both proteins.
Results The positive particles of RARRES3 and MIEN1 were located in the cytoplasm. The expression of RARRES3 in NSCLC tissues was 29.07％ （25/86）， lower than 55.56％ （40/72） of para-carcinoma tissues （P＜0.05）， while the expression of MIEN1 in NSCLC tissues was 62.79％ （54/86）， higher than 31.94％（23/72） of para-carcinoma tissues （P＜0.05）. The expression of RARRES3 and MIEN1 protein in NSCLC was related to TNM stage and lymph node metastasis. Moreover， RARRES3 was also associated with tumor size， and MIEN1 protein was also associated with differentiation type （P＜0.05）. RARRES3 protein expression were negatively correlated with MIEN1 （r=-0.411，P=0.000）. The AUC， sensitivity and specificity of RARRES3 and MIEN1 in NSCLC were 0.825 and 0.779， 85.1％ and 75.4％， 79.2％ and 83.3％， respectively. The median overall survival （OS） was 32.5 months in the RARRES3 high expression group， which was higher than 16.0 months in the low expression group， and the median OS was 15.5 months in MIEN1 high expression group， which was lower than 28.0 months in the low expression group （P＜0.05）. Conclusion In NSCLC tissues， RARRES3 was lowly expressed and MIEN1 was highly expressed. Both proteins were related to TNM staging， lymph node metastasis and prognosis. It may be related to the occurrence and development of NSCLC.

Objective To explore the expression of Toll like receptor-9（TLR-9）and its correlation with microvessel density（MVD）and vascular endothelial growth factor（VEGF）in cervical squamous cell carcinoma. Methods The expression of TLR-9 was detected by immunohistochemical SP method in 48 cases of cervical squamous cell carcinoma and 20 cases of normal cervical epithelium. The tissues were labeled with CD34 monoclonal antibody to calculate MVD， and the level of VEGF was measured by enzyme linked immunosorbent assay. The effectiveness of TLR-9 expression in the diagnosis of cervical squamous cell carcinoma was evaluated by receiver operating characteristic curve. Results In the tissues from cervical squamous cell carcinoma， the positive expression rate of TLR-9 was 83.3％（40/48）， and the MVD and level of VEGF were 36.8±7.25 and（25.58±3.49）ng/mg， all higher than those of normal cervical epithelial tissues（P＜0.05）. The high expression of TLR-9 was related to the clinical stage and the expression of matrix metalloproteinase 9， but unrelated with histological grade and lymph node metastasis. The levels of VEGF and MVD in tissues with TLR-9 positive expression were significantly higher than those in the negative group（P＜0.05）. The diagnostic efficiency of TLR-9 in cervical squamous cell carcinoma was good， and its AUC， sensitivity and specificity were 0.830， 76.1％ and 82.7％， respectively. Conclusion The carcinogenesis and metastasis with high expression of TLR-9 may be involved in cervical squamous cell carcinoma， and angiogenesis is closely related with cervical squamous cell carcinoma.

Objective To investigate the expression of HOXB9 in gastric cancer tissues and its correlation with clinicopathological features， and to analyze its prognostic value in patients with gastric cancer. Methods Immunohistochemical SP method was used to detect the expression of HOXB9 in 190 gastric cancer tissues and adjacent normal tissues. And the correlation of its expression with clinicopathological features and prognosis was studied. Results The positive rate of HOXB9 was 95.8％（182/190） and 45.3％（86/190） in adjacent normal tissue and gastric cancer tissue（P＜0.001）. The expression of HOXB9 was associated with histological differentiation， TNM stage and lymph node metastasis，but not with gender and age. The median overall survival（OS） of gastric cancer patients was 31 months， and the patients with HOXB9 positive expression was 26.0 months， 1ower than 36.0 months of patients with negative expression（P＜0.005）. Cox proportional hazards model revealed that HOXB9 expression was an independent prognostic factor for postoperative survival of gastric cancer. Conclusion HOXB9 may be involved in the occurrence and development of gastric cancer. HOXB9 may serve as an prognostic biomarker for the survival of patients with gastric cancer.

Objective To investigate the expression level of Septin 9 isoform 1（SEPT9_i1）and promyelocytic leukaemia zinc finger（PLZF）protein in gastric cancer tissues， and to analyze the relations between them with the clinical pathological features and prognosis of gastric cancer. Methods The immunohistochemistry EnVision two step method was used to detect the SEPT9_i1 and PLZF expression in paraffin embedded 75 cases of gastric cancer tissues and 62 cases of paracancerous tissue. The staining Results of the tumors were expressed as the product of the staining intensity and the percentage of positive tumor cells. The expression was divided into high expression group（＜5）and low expression group（≥5）. We analyzed the correlation between tumor tissue expression and clinical parameters， such as gender， age， clinical stage， tumor location， Lauren typing， tumor size， differentiation， depth of invasion and lymph node metastasis. Moreover， the correlation between tumor tissue expression and prognosis were also analyzed. The Spearman correlation analysis was used to analyze the expression of PLZF and SEPT9_i1 in gastric cancer tissues. Results The higher expression of SEPT9_i1 in gastric cancer tissues was higher than that in para-carcinoma tissues（65.33％ vs. 37.10％， P＜0.05）， and the PLZF expression was lower than that in para-carcinoma tissues（41.33％ vs. 85.48％， P＜0.05）. There was negative correlation between expression of SEPT9_i1 and PLZF in gastric cancer tissues（P=0.009， r=-0.299）. The expression of both proteins was related to clinical stage and lymph node metastasis. Expression of SEPT9_i1 was related to the depth of invasion， and the expression of PLZF was related to the differentiation degree（P＜0.05）. The median overall survival（OS）was 21.5 months in patients with high expression of SEPT9_i1， lower than 32.0 months of patients with low expression of SEPT9_i1（P＜0.05）. The median OS was 26.5 months in patients with high expression of PLZF， similar to 31.0 months in patients with low expression of PLZF（P＞0.05）. Conclusion SEPT9_i1 was highly expressed in gastric cancer tissues but PLZF was low expression. Both were related to clinical stage and lymph node metastasis, and SEPT9_i1 expression is related to the prognosis of gastric cancer patients，suggesting that they may play an important role in the occurrence and development of gastric cancer，and can be used in the diagnosis and evaluation of gastric cancer.

Objective To investigate the expression of latexin（LXN） in gastric cancer and determine the relationship of its expression with clinical features and prognosis. Methods The expression level of LXN was detected by tissue microarray technology and immunohistochemistry in 143 cases of gastric cancer and paired adjacent normal tissues. The relationship between the expression of LXN and clinical parameters were analyzed as well as overall survival（OS）. Results The positive rate of LXN in gastric cancer tissues was 60.1％（86/143）， lower than 100.0％（143/143） of para-carcinoma tissues（P＜0.05）. A significant correlation was observed between the expression of LXN and N stage（P=0.005）， tumor differentiation（P=0.023）， but not with age， gender， tumor size， T stage and TNM stage. The medium OS of 143 patients was 56.0 months，and for the patients with LXN negative expression was 27.0 months， shorter than 79.5 months of LXN positive expression（P＜0.05）. Multivariate analysis revealed that tumor size was the independent prognostic factor in gastric cancer patients（P=0.018）. Conclusion LXN is down-regulated in gastric cancer and may relate to tumorgenesis and development. Loss of LXN expression is associated with the poor prognosis of gastric cancer.

Objective To investigate the expression and significance of RIPK4 protein in human osteosarcomatous tissue and cells. Methods Imunohistochemistry was performed to detect the expression of RIPK4 protein in paraffin speciments of 30 osteosarcoma patients（and 10 osteochondroma patients. The correlation among clinicopathologic characteristics and RIPK4 protein was analyzed. The expression of RIPK4 mRNA in osteosarcoma MG-63， U-2OS and osteoblast cells was detected by RT-PCR， and differences were compared. Results The positive expression of RIPK4 in osteosarcoma was 60.0％（18/30）， while it was 20.0％（2/10） in osteochondroma.The differences were statistically significant（P＜0.05）. RIPK4 mRNA of osteoblast cells was 0.6500±0.2121， which was significantly lower than that of MG-63（1.6079±0.2661） and U-2OS（1.5000±0.1414），with singnificant differences（P＜0.05）. The expression of RIPK4 protein was related to TNM stage and lung metastasis（P＜0.05）. Conclusion The expression of RIPK4 protein and gene in osteosarcoma tissues and cells is high. RIPK4 may be involved in the progress of epithelial osteosarcoma.

Objective To investigate the relationship between helicobacter pylori（Hp） infection and expression level of survivin mRNA in the peripheral blood of gastric cancer patients. Methods The expression levels of survivin mRNA in the peripheral blood of gastric cancer and gastritis were detected by real-time quantitative reverse transcription polymerase chain reaction （qPCR）. Hp infection was determined by rapid urease test and giemasa staining. Results In the specimens of gastritis patients and gastric cancer patients，the expression rates of survivin mRNA were 0 and 84.0％. Median value was 0.46 for expression of survivin mRNA in the peripheral blood of gastric cancer patients. The expression levels of survivin mRNA were unrelated with age， gender and lymph node metastasis（P＞0.05）， but was significantly related with TNM stage（P＜0.05）. The positive expression rate of Hp in gastric cancer was 80.0％（40/50）， significantly higher than 58.0％（29/50） in gastritis patients（P＜0.05）. In the gastric cancer patients, the expression levels of survivin mRNA in Hp postive infection was 0.578±0.507， higher than 0.194±0.419 in Hp negative infection with significant differences （P＜0.05）. The expression survivin mRNA in gastric cancer patients and Hp infection was positively correlated（r=0.464， P＜0.05）. Conclusion The expression of survivin and Hp infection may be correlated with gastric cancer. Survivin may participate in the progress of gastric cancer with Hp infection.

Objective To explore the expression and clinical significance of lncRNA CADM1-AS1 in lung adenocarcinoma. Methods In this study, 72 cases of lung adenocarcinoma tissues and corresponding para carcinoma tissues were collected from our hospital. Real time quantitative PCR （qPCR） was used to detect the level of CADM1-AS1 in tissue samples. The relationship between CADM1-AS1 level and clinical pathological parameters （gender， age， tumor size， differentiation， clinical stage， T stage， smoking history and lymph node metastasis） was analyzed. The median overall survival （OS） was compared between different CADM1-AS1 levels according to the long-term survival data. The levels of CADM1-AS1 in common non-small cell lung cancer cells （NCI-H1299， SPC-A， NCI-H1650 and A549） were detected by qPCR assay. The effectiveness of CADM1-AS1 level in early diagnosis of lung cancer was evaluated by the receiver operating characteristic curve （ROC）. Results The level of CADM1-AS1 in lung adenocarcinoma tissues was lower than that in para carcinoma tissues （P＜0.05）. Compared with the normal bronchial epithelial cell line 16HBE，the CADM1-AS1 of the non small cell lung cancer was lower （P＜0.05）. CADM1-AS1 level was related to TNM stage， differentiation， tumor size， T stage and lymph node metastasis of lung adenocarcinoma （P＜0.05）. The median OS in patients with high CADM1-AS1 level was 32.5 months， higher than 16.0 months of patients with low CADM1-AS1 level （P＜0.05）. The value of CADM1-AS1 in the early diagnosis of lung adenocarcinoma was good with the area under the curve of 0.874， and the sensitivity and specificity were 87.8％ and 85.2％ respectively. Conclusion There were relative low levels of CADM1-AS1 in lung adenocarcinoma cell line and non-small cell lung cancer cell line. The CADM1-AS1 level is related to the TNM stage， differentiation and prognosis， which may be related to the occurrence and development of lung adenocarcinoma. It has a good value for early diagnosis of lung adenocarcinoma.

Objective To assess the effect of optimizing the neck node level Ⅳ in IMRT clinical target volume（CTV）for nasopharyngeal carcinoma（NPC）on the treatment outcome and thyroid dose， in order to provide the basis for optimization of IMRT target area in NPC. Methods For selected 64 cases of NPC patients treated with IMRT，including 32 cases of neck node level Ⅳ region which optimized，and a control group of 32 cases which Ⅳ region is still within the CTV；Radiation dose parameters of thyroid were measured using treatment plan system for all patients. These radiation dose parameters of thyroid were compared between optimization and no optimization groups，as well as among different clinical stages， T and N stages， respectively. Meanwhile， a survival analysis was performed using Kaplan-Meier method. Results Reasonably optimizing the neck node level Ⅳ in CTV can significantly reduce the thyroid mean dose and volume percent dose V50（P＜0.05） and does not decrease the local control probability. The thyroid mean dose and V50 of patients with stage Ⅳ were much higher than those with other stages， and for the N0-2 patients， optimization of neck node level Ⅳ in CTV can significantly reduce the thyroid mean dose and V50 comparing with no optimization group. Conclusion For NPC patients， reasonably optimizing the neck node level Ⅳ in CTV is safe and can much better protect the thyroid gland.

Objective To evaluate clinical efficacy and prognosis of gefitinib and erlotinib against brain metastases of non-small cell lung cancer（NSCLC）and to compare the difference between two drugs. Methods A total of 67 brain metastases patients of pulmonary adenocarcinoma harboring activating EGFR mutation were reviewed retrospectively. All of them were treated with oral either gefitinib（250 mg/day, n=38）or erlotinib（150 mg/day, n=29）. These patients discontinued administration of gefitinib or erlotinib when intracranial disease progression， death or intolerable side effects appeared. RECIST 1.1 was applied in response analysis. Survival analysis was compared with Kaplan-Meier method and Log-rank test respectively. Results In terms of intracranial diseases， response rate（RR）and disease control rate（DCR）were 44.8％ and 92.5％， respectively. Furthermore， RR and DCR were 42.1％， 92.1％ and 48.3％，93.1％ in gefitinib and erlotinib group， respectively（P=0.881）. As for extracranial diseases， RR and DCR were 53.7％ and 95.5％， respectively. In addition， RR and DCR were 52.6％， 94.7％ and 55.2％， 96.6％ in gefitinib and erlotinib group， respectively（P=0.932）. The median progression free survival（PFS）and overall survival time（OS）was 10.8 months and 15.3 months， respectively. Furthermore， median PFS and OS was 10.6 months， 14.8 months and 11.7 months， 15.7 months in gefitinib and erlotinib group， respectively（P=0.720， P=0.569）.
Conclusion Gefitinib and erlotinib showed promising antitumor activity against brain metastases in NSCLC patients harboring activating EGFR mutation and appear to be the treatment of choice in this clinical setting. Gefitinib and erlotinib seem to show no difference in efficacy and prognosis of these patients.

Objective To investigate the value of PET-CT image fusion to delineate the target of intensity modulated radiation therapy（IMRT） in locally advanced non-small cell lung cancer（NSCLC）， and the impact on target volume and dose of normal lung tissue. Methods Thirty NSCLC patients of clinical stage ⅢA and ⅢB were randomly selected. Target and organ at risk were delineated on the same fixed position according to enhanced CT images and fusion images of PET/CT and CT, respectively. Then the volume of gross tumor volume（GTV） and planning target volume（PTV） under these two status were compared. Moreover， the percent of the total lung volume exceeding 5 Gy（V5）， percent of the total lung volume exceeding 20 Gy（V20）， mean dose of lung irradiated（MLD） in two different conformal IMRT plans were observed when the dosage of PTV was up to 60 Gy/30 f. Results The GTV volume on fusion images of PET-CT/CT was （248.39±94.80）cm3, less than （311.22±99.16）cm3 on the enhanced CT images in 30 cases. The difference was statistically significant（P＜0.05）. PTV， extended from GTV， in fusion images of PET-CT/CT was （356.68±92.73）cm3, while in the enhanced CT images were （433.58±107.89 cm3） with significant difference（P＜0.01）. The V5， V20 and MLD of whole lung in the two plans were compared. In fusion images of PET-CT/CT program， V5， V20 and MLD was （51.26±10.50）％，（25.71±5.17）％ and （1595.27±148.24） cGy, remarkly less than （56.41±9.55）%, （29.09±4.10）% and （1693.59±100.60） cGy in the enhanced CT program with significant difference（P＜0.05）. Conclusion Application of PET-CT/CT fusion image to delineate the targets of IMRT can improve the accuracy of target delineation volume and reduce the dose of normal lung tissues.

Objective To explore the clinical effect of the radiofrequency ablation （RFA） in combination with S-1 and oxaliplatin （SOX regimen） for colorectal cancer with liver metastases. Methods Among 44 patients with colorectal liver metastasis from April 2011 to April 2013， 21 cases received RFA in combination with SOX regimen （combined treatment group） and other 23 cases only received SOX regimen （chemotherapy alone group）. The curative effect of chemotherapy and RFA was analyzed according to Response Evaluation Criteria in Solid Tumors （RECIST） 1.1 or three-phase enhanced CT， respectively. The long-term survival of the patients was followed up， and the complications of RFA and the chemotherapy-induced adverse reactions were recorded. Results All the 44 patients were evaluable for response. In combined treatment group， there were 9 cases of CR， 6 cases of PR， 3 cases of SD and 3 cases of PD with the response rate （RR） and disease control rate（DCR） of 71.4％ and 85.7％. In chemotherapy alone group， there were 2 cases of CR， 6 cases of PR， 5 cases of SD and 10 cases of PD with RR and DCR of 34.8％ and 6.5％. There were higher DCR and RR in combined treatment group versus chemotherapy alone group （P＜0.05）. Among the 31 lesions receiving RFA， 19 lesions （61.3％） were completely destroyed and 13 lesions （41.9％） were locally recurred. The survival rates of 1-， 2-， 3-year were 71.4％ （15/21）， 52.4％ （11/21） and 33.3％ （7/21） in combined treatment group， higher than 26.1％ （6/23）， 21.7％ （5/23） and 8.70％ （2/23） in chemotherapy alone group （P＜0.05）. The main complications of RFA were fever and local pain. There were similar adverse reactions in both groups， mainly including bone marrow suppression and gastrointestinal reaction， and the difference had not statistical significance （P＞0.05）. Conclusion RFA in combination with systemic chemotherapy treatment of colorectal liver metastasis can effectively control the tumor and prolong the patients survival.

Hepatocellular carcinoma （HCC） is one of the most common malignancy tumors in the world， which is mainly caused by Hepatitis B virus （HBV） infection. HBV DNA integration may promote tumorigenesis via cis-activation of oncogenes or altered tumor suppressor activity, genomic destabilization and production of mutant HBV proteins with tumor-promoting potential. The X gene product of HBV （HBx） has extensive trans-activating functions， which can regulate cell apoptosis， suppress cell DNA repair， intervene intracellular signaling pathways and change epigenetic. In this review， we summarized the research progress of prevalence and incidence of HBV related HCC， HBV DNA integration and HBx protein.

Lung cancer is the leading cause of cancer deaths throughout the world. Non-small cell lung cancer （NSCLC） is the most common type of lung cancer. However， its pathogenesis has not been elucidated. Recently， researches have shown that p44/WDR77 expression is essential for the growth of lung epithelial cells and orthotropic lung adenocarcinoma tumors. The expression of 44-KDa androgen receptor cofactor/WD repeat domain 77 （p44/WDR77） gene in NSCLC is significantly higher than that in adjacent tissues. Loss of p44/WDR77 expression in lung epithelial cells and lung adenocarcinoma cancer cells leads to G1 cell cycle arrest via the p21-Rb-E2F signaling pathway. P44/WDR77 was re-expressed in lung cancer， and silencing p44/WDR77 expression strongly inhibited growth of lung adenocarcinoma tumor xenografts in mice. This article will briefly summarize the research progresses of p44/WDR77 in NSCLC.