Objective: To study the prognostic significance of the subtype of SYT-SSX fusion gene, E-cadherin, β-Catenin and clinicopathologicel parameters for the metastasis of synovial sarcomas. Methods: A total of 98 synovial sar-coma patients with complete clinical and follow-up data were reviewed. RT-PCR was used to detect the subtype of SYT-SSX fusion geneo The expression of E-cadherin and β-catenin was detected by immunohistochemistry. Univariate and multivariate analyses were performed to analyze the influence of the above factors and clinicopathological parameters on the metastasis free survival to explore the factors affecting the metastasis of synovial sarcoma. Results: Of all the pa-tients, 69.4% (68/98) had metastasis during follow-up. The median metastasis free survival was 48 months. The metastasis free 1-, 2-, 3-, 4-, and 5-year survival rate after surgery was 97.5%, 75.5%, 63.5%, 54.0%, and 48.5%, respectively; 31.6% (31/98) patients were found with SYT-SSX1 and 68.4% (67/98) patients with SYI-SSX2. The positive rate of E-cadherin ex-pression was 38.8% (38/98), the positive rate of β-catenin expression was 39.8% (39198) on cellular membrane and 53.1% (52/98) in cellular nucleus/cytoplasm. Univariate analysis showed that age (P=0.003), mitotic figure (P=0.002), histological grade (P=0.001), the subtype fusion gene of SYT-SSX (P=0.014), E-cadherin expression (P=0.015) and β-catenin expres-sion on cellular membrane (P=0.020) were significantly correlated with metastasis free survival of synovial sarcoma pa-tients. Sex (P=0.190), tumor location (P=0.105), tumor size (P=0.180), histological type (P=0.354), necrosis (P=0.451), β-catenin expression in cell nucleus/cytoplasm (P=0.911), radiotherapy (P=0.193), and chemotherapy (P=0.249) had no sig-nificant correlation with metastasis free survival of synovial sarcoma patients. Multivariate analysis revealed that the sub-type of SYT-SSX1 fusion gene (RR=2.505, P=0.003), negative expression of E-cadherin (RR=3.282, P=0.000), patient age (RR=2.157, P=0.004), and grade Ⅲ (RR=1.784, P=0.030) were independent risk factors for metastasis of synovial sarco-ma. Conclusion: The subtype of SYT-SSX, expression of E-cadherin, histological grade and the age of patients are impor-tant factors for evaluating the metastasis and prognosis of synovial sarcoma.

Objective: To study the prognostic significance of cell proliferation and apoptosis, MVD and clinicopathologi-cal parameters for the recurrence of synovial sarcoma. Methods: We analyzed the clinical and follow-up data of 56 synovial sarcoma patients without metastasis. RT-PCR was used to detect the subtype of SYT-SSX fusion gene. The expression of Ki67 and MVD was detected by immunohistochemistry. Univariate analysis was employed to analyze the influence of the above factors and clinicopathological parameters on the recurrence free survival and to explore the influencing factors for the recurrence of synovial sarcoma. Results: Of all the patients, 73.2% (41/56) had recurrence during the follow-up. The median recurrence free survival was 19.5 months. The recurrence free 1-, 2-, 3-, 4-, and 5-year survival rates after surgery were 45.0%, 41.0%, 34.0%, 28.0%, and 28.0%, respectively. Ki-67 labeling index (LI) was 19.98%±11.64% and MVD was 51.83±21.92 per ×400. There was no significant difference in apoptotic index (AI) between the two groups (P=0.607). Χ~2 analysis showed that histological type (P=0.000) and MVD (P=0.045) were significantly correlated with the recurrence of sy-novial sarcoma. Univariate analysis showed that Ki67 LI (P=0.009), histological type (P=0.012) and radiotherapy (P= 0.014) were significantly correlated with the recurrence free survival of synovial sarcoma patients. Sex (P=0.015), tumor lo-cation (P=0.411), tumor size (P=0.801), necrosis (P=0.486), MVD (P=0.454), chemotherapy (P=0.272), and apoptotic grade (P=0.899) were not correlated with the recurrence free survival of synovial sarcoma patients. Multivariate analysis re-vealed that higher expression of Ki67 (RR=1.944, P=0.045), radiotherapy (RR=0.482, P=0.04), and histological type (RR= 0.207, P=0.031) were independent risk factors for the recurrence of synovial sarcoma. Conclusion: The expression of Ki67, radiotherapy and histological type are important factors for evaluating the recurrence and prognosis of synovial sarcoma.

Objective: To investigate the impact of lincRNA-ROR, a ceRNA by binding miR-145 on the expression of the downstream genes Oct4, Sox2 and Nanog, and related biological characteristics of colon cancer stem cells, and to elucidate the clinical significance of this molecular regulatory network. Methods: Fifty-two cases of colorectal cancer tissue and adjacent tissue were collected at Nanyang City Central Hospital and Nanyang Second Hospital, Henan Province, from 2014 to 2016. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression of lincRNA-ROR and miR-145 in colorectal cancer tissue and isolated colon cancer cells. The correlation between the expression of lincRNA-ROR, miR-145 and the clinicopathologic features of colon cancer was performed. CD44^-CD133^- and CD44⁺ CD133⁺ cells were isolated from SW1116 by using flow cytometry. The expression of CD44, CD133, Oct4, Sox2, Nanog, lincRNA-ROR and miR-145 in cells were detected by qPCR. The relationship between lincRNA-ROR, miR-145, Oct4, Sox2 and Nanog was analyzed by bioinformatics, dual luciferase reporter assay, qPCR and Western blot. The effects of silencing lincRNA-ROR on the proliferation and chemosensitivity of colon cancer stem cells were detected by MTT, colony formation. Results: LincRNA-ROR was frequently up-regulated and inversely correlated with miR-145 down-regulation in the colon cancer specimens(P<0.05). LincRNA-ROR was related to tumor size, lymph node involvement and distant metastasis(P<0.05), and miR-145 was found related to tumor size and tumor location(P<0.05). CD44⁺ CD133⁺ cells were successfully isolated from SW1116 by flow cytometry. The levels of CD44, CD133, Oct4, Sox2, Nanog, lincRNA-ROR in CD44⁺ CD133⁺ cells were significantly increased, while miR-145 was decreased compared with CD44^-CD133^-cells(P<0.05). The levels of CD44, CD133, lnc-ROR in CD44⁺ CD133⁺ cells were significantly reduced upon cell adherence, while miR-145 was significantly increased(P<0.05). Bioinformatics analysis revealed that lincRNA-ROR shared miRNA response elements with core transcription factors Oct4, Sox2 and Nanog. MiR-145 significantly inhibited the expression of lincRNA-ROR, Oct4, Sox2 and Nanog. Silencing lincRNA-ROR significantly inhibited colon cancer stem cells proliferation and increased the sensitivity to chemotherapy. Conclusions Linc-ROR functions as a key ceRNA to prevent core TFs, e. g., Oct4, Sox2, Nanog, from miR-145-mediated suppression in colon cancer stem cells and regulates cell proliferation and chemosensitivity.The data may provide insights into the pathophysiological interactions of the components of genetic networks in the development of colon cancer and may lead to new therapies in the future.

Objective: To investigate differentially expressed protein profiles in B16-F10 grafted melanoma and its metastasis in the lung in order to identify molecular markers of melanoma metastasis. Methods: Differentially expressed proteins in B16-F10 grafted melanoma and its metastatic lesion in the lung were isolated and identified by fluorescence two-dimensional differential gel electrophoresis(2D-DIGE)coupled with matrix assisted laser desorption ionisation time-of-flight mass spectrometry(MALDI-TOF-MS).Some of identified proteins were further confirmed by Real-time PCR analysis. Results: High resolutional images of differential gel electrophoresis were obtained and 9 of 30 differentially expressed proteins (IRatiol≥2,P<0.01)were identified by MALDI-TOF-MS.The expression of Myoglobin(MB),vimentin(VIM),phosphoglycerate kinase 1(PGK1),Triosephosphate isomerase(TPI or TIM),heavy-chain binding protein(BiP),α-enolase,β-actin,γ-actin,and laminin-binding protein were up-regulated in the experimental group compared with the control group.These proteins were involved in the cytoskeletal formation,glycolysis and so on.Real-time PCR analysis showed up-regulation of mRNA expression of PGK1 and TPI in the experimental group(P=0.001 and 0.003),which was in consistent with the resuits of proteomic analysis. Conclusion: A variety of abnormally expressed proteins contribute to the metastasis of mice melanoma.Glycolytic enzymes PGK1 and TPI may be involved in this process.

Objective: To study the relationship of LPPCN with neovascularization and to analyze its clini-copathologic significance, in an effort to find a new target for anti-vascular therapies. Methods: Sixty-eight ma-lignant melanoma specimens were analyzed to observe the distribution of LPPCN and to examine the expres-sion of CD105 and TGFβ1 using immunohistochemistry. The distribution of vasculogenic mimicry (VM) was observed by immunohistochemical and histochemical double staining of CD31 and PAS. Results: (1) The tu-lines and networks. Of the 68 cases of melanoma, 55.89% (38/68) were recognized as having LPPCN. (2) In malignant melanoma specimens, the rate of vasculogenic mimicry density (VMD) and microvessel density (MVD) labeled by CD105 in LPPCN-positive group were higher than those in LPPCN-negative group, with sig-nificant differences (P<0.05). VMD and MVD were positively-correlated with the density of LPPCN. The posi-tive expression of TGFβ1 in LPPCN-positive group was higher than that in LPPCN-negative group and its ex-pression in the regions of LPPCN was obviously higher than that in circumambient tumor cells, with a signifi-cant difference (P<0.05). The expression of TGFβ1 was positively correlated with MVD labeled by CD105. (3) There was no relationship between LPPCN and gender, age, site, tumor embolus, lymph node metastasis or distant metastasis (P>0.05), but LPPCN was correlated with tumor size, mitosis figure count and Breslow depth (P<0.05). Kaplan-Meier survival analysis showed the survival rate of patients with LPPCN was lower than that of patients without LPPCN, with a statistical significance (P<0.05). The presence of LPPCN indicat-ed poor prognosis. Conclusion: LPPCN exists in malignant melanoma and is associated with VM and angio- genesis. Some tumor cells undergoing LPPCN have a spacial foundation for VM and angiogenesis. LPPCN can be an index for the evaluation of the prognosis of melanoma patients.

Objective To measure the value of orthopedic physiological and operative severity score for the enumeration of mortality and morbidity (POSSUM) and Portsmouth modified POSSUM (P-POSSUM) scoring systems in predicting operative risks in aged hip fracture patients.Methods Orthopedic POSSUM and P-POSSUM were performed to predict complication incidence and mortality for 164 aged patients operated for hip fracture.Validation of the scoring systems was tested by assessing observed to expected ratio,discrimination,and calibration.Discriminative ability and calibration of both scores were estimated using receiver operation characteristic curve (ROC) and Hosmer-Lemeshow test respectively.Results Orthopedic POSSUM score performed in predicting incidence of postoperative complications showed overall observed to expected ratio of 0.86,area under the curve of 0.82,and good calibration (H2 =3.66,df=8,P ＞ 0.05).P-POSSUM performed in predicting mortality showed overall observed to expected ratio of 0.80,area under the curve of 0.93 and good calibration (H2 =3.21,df =4,P ＞ 0.05).While orthopedic POSSUM overestimated postoperative mortality (overall observed to expected ratio =0.27).Conclusion Orthopedic POSSUM and P-POSSUM scores are respectively accurate in predicting postoperative complication incidence and mortality in aged hip fracture patients,but orthopedic POSSUM score overestimates the mortality.

Objective To analyze the expression of Kruppel-like factor 4 (KLF4 ) and CD44 in esophageal squamous cell carcinoma (ESCC) tissues and adjacent non-cancerous tissues ,and to investigate their effects on the prognosis .Methods From June 2012 to September 2016 ,in The Second People′s Hospital of Nanyang ,a total of 100 patients with ESCC who receiving operation were selected .The ESCC tissues and the adjacent non-cancerous tissues (control) of the patients were collected .The expression levels of KLF4 and CD44 protein were detected by immunohistochemistry .The expressions of KLF4 and CD44 at mRNA and protein level of 50 paired fresh tissues were examined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting ,respectively . T-test ,chi-square ,Kaplan-Meier method and Pearson correlation analysis were performed for statistical analysis .Results The positive expression rate of KLF4 protein in ESCC tissues was 55％ (55/100) ,which was lower than that in adjacent non-cancerous tissues (77％ ,77/100) ,and the difference was statistically significant (χ2 =10 .778 ,P=0 .001) .The positive expression rate of CD44 protein in ESCC tissues was 81％ (81/100) ,which was higher than that in adjacent non-cancerous tissues (11％ ,11/100) ,and the difference was statistically significant (χ2=112 .600 ,P<0 .01) .The expression level of KLF4 mRNA in 43 cases was lower than that in adjacent non-cancerous tissues ,the expression level of CD44 mRNA in 46 cases was much higher than that of adjacent non-cancerous tissues ,and the differences were statistically significant (χ2 =51 .837 and 70 .563 ,both P< 0 .01) .There were statistically significant differences in positive expression rates of KLF4 in cancer tissues between different gender , differentiation degree , invasion depth ,TNM stage and lymph node metastasis (all P<0 .05) .Similarly there were statistically significant differences in positive expression rates of CD 44 in cancer tissues between different differentiation degree ,invasion depth ,TNM stage and lymph node metastasis (all P< 0 .05) .The expression of KLF4 was negatively correlated with CD44 expression ,either at protein level or mRNA level (r= -0 .284、-0 .518 ,both P< 0 .01) .The median survival time of patients with positive KLF4 expression in cancer tissues was 33 months ,which was longer than that of patients with negative KLF4 expression (20 months) ,and the difference was statistically significant (χ2 =4 .021 , P= 0 .019) .The median survival time of patients with positive CD44 expression in cancer tissues was 24 months ,which was shorter than that of patients with negative CD44 expression (37 months) , and the difference was statistically significant (χ2 = 4 .379 , P= 0 .016) .The results of univariate analysis showed that TNM stage ,KLF4 expression and CD44 expression were correlated with overall survival time (all P<0 .05) . The results of multivariate analysis indicated that TNM stage ,lower KLF4 expression and higher CD44 expression were the independent risk factors for survival (all P<0 .05) .Conclusion Lower expression of KLF4 and higher expression of CD44 in ESCC may be closely correlated with its occurrence ,development and prognosis .

Objective To investigate the expression levels of serum miR-210and miR-375in patients with non-small cell lung cancer (NSCLC).Methods A total of 25NSCLC patients (NSCLC group) and 14healthy volunteers (control group) were enrolled in this study.The relative expression levels of 9miRNAs (miR-182, miR-126, miR-31, miR-21, miR-221, miR-200b, miR-183, miR-210and miR-375) in 6 NSCLC patients and 6healthy volunteers were measured by RT-qPCR.The dysregulated miRNAs will be selected as candidate miR-NAs.The diagnostic value were evaluated by ROC curve.Results Compared with control group, 2 (miR-210and miR-375) out of 9miRNAs were up-regulated in NSCLC group, and the differences were statistically significant (P＜0.05), while the other 7miRNAs were not consistent with the reported literatures.Therefore, miR-210and miR-375were selected as candidate miRNAs.We found that the relative expression level of miR-210in the lung adenocarcinoma group was significantly different from control group (P＜0.05), while there was no significant difference between the squamous cell carcinoma group and the control group (P＞0.05).There was no significantly statistical difference in the relative expression level of miR-375between lung squamous cell carcinoma group, lung adenocarcinoma group and the control group (P＞0.05).The AUC of serum miR-210of lung adenocarcinoma group was 0.737 5 (95％CI:0.498 3-0.976 7, P=0.091 4) with a medium diagnostic value.Conclusion MiR-210is highly expressed in the serum of patients with lung adenocarcinoma, suggesting that miR-210may be a novel tumor marker for the diagnosis of lung adenocarcinoma.The value of miR-375in the diagnosis of lung cancer still needs to be further explored.