Objective To detect biomarker proteins in relation to metastasis by comparing serum protein profiles of primary colorectal cancer patients with or without metastasis. Methods A total of 219 serum samples were analyzed using surface enhanced laser desorption ionization time of flight mass spectrometry(SELDI-TOF-MS).The samples were divided into two groups:the training group consisting of 57 primary colorectal cancer patients、63 metastatic colorectal cancer patients and 42 healthy controls,and the test group consisting of 26 primary colorectal cancer patients and 31 metastatic colorectal cancer patients.Samples in training group were analyzed to identify serum biomarker proteins which could differentiate colorectal cancer patients with or without metastasis.The sensitivity and specificity of biomarker proteins were examined by results from blind test group. Results In the m/z region of 2000～30000,31 proteins had statistically significant difference between primary colorectal cancer patients and healthy controls.The m/z of difierentiated proteins were respectively 3240.7、9289.3、5334.4、4596.1 and 4792.4 according to P value.38 proteins had statistically significant diffefence between metastatic colorectal cancer patients and healthy controls.The m/z of differentiated proteins were respectively 3240.7、9289.3、9184.4、3191.5 and 9340.9 according to P value.Only two protein peaks(9184.4 and 9340.9)were found of statistical difference between primary eolorectal cancer patients and metastatic colorectal cancer patients.The sensitivity and specificity of the combination use of the two biomarkers were respectively 90.3% and 88.5% in the test group. Conclusion SELDI-TOF-MS was helpful to find protein biomarkers with relation to metastasis in colorectal carcinoma patients.

ObjectiveTo investigate emergency diagnosis and treatment of pelvic fracture complicated with traumatic rupture of urethra and bladder,and to improve the success rate of treatment on pelvic fracture.MethodsClinical data of 52 cases of pelvic fracture complicated with traumatic rupture of urethra and bladder in department of emergency and urology from 2000 to 2010 was retrospectively analyzed.Results Among the 52 patients,there was 41 cases of pelvic fracture complicated with posterior urethral disruption,15 cases complicated with rupture of bladder and 4 cases complicated withtraumatic rupture of urethra and bladder at the same time.In 41 cases with posterior urethral rupture,6 individual's condition were relatively so severe that they onlyunderwent bladder puncture nephrostomy,and 29 cases underwent traction urethral realignment,the other 6 cases didn't undergo surgery; In 15 cases of patients with bladder rupture,2 patients were performed urethral realignment and bladder repair,11 patients underwent the bladder repair only and the other 2 patients were not performed surgery.There were 8 patients died and the mortality rate was 15.4％.Six died cases failed to conduct emergency surgery because of uncontrollable bleeding and another 2 cases died due to multiple organ failure.ConclusionPelvic fractures is a disease with more complications,it should be diagnosed as early as possible.Patients invalid for conventional anti-shock should be performed pelvic external fixation and emergency embolization to stop bleeding in the emergency department,and undergo associated processing after they are in stable condition.

Objective To establish a direct reverse transcription real-time fluorescence quantitative polymerase chain reaction ( RT-qPCR-D ) method for detecting serum circulating B cell-specific moloney murine leukemia virus integration site-1 (Bmi-1) mRNA, and analyze the levels of serum circulating Bmi-1 mRNA in colorectal cancer patients by using of this method for exploring its diagnosis value in colorectal cancer.Methods Methodology establishment.RNA was extracted from colorectal cancer HT 29 cell line, and detection standard curves of Bmi-1, ubiquitin C ( UBC), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) mRNAs were established , then the amplification efficiencies were calculated.Bmi-1 mRNA level was directly detected in serum and preparation buffer mixture , then the specificity of assay was evaluated by melting curve, and detection limit was observed through diluted serum samples.The serum circulating Bmi-1 mRNA levels were detected by ELISA in 158 cases with colorectal cancer , of which there were 26 cases of tumor node metastasis ( TNM)Ⅰstage, 53 cases of TNMⅡ, 47 cases of TNMⅢ, 32 cases of TNMⅣand 53 cases of controls with normal colonoscopy collected from January 2008 to January 2009 in Qilu Hospital of Shandong University.Comparisons of groups were determined by applying Mann-Whitney U test or Kruskal-Wallis test, and receiver operating characteristic ( ROC) curves were established to illustrate the diagnostic performance.Results The log values of Bmi-1, UBC and GAPDH showed good linear correlations with quantification cycle (Cq) values(R2 =0.990, 0.990, 0.991, all P <0.001), and the amplification efficiencies were 0.875, 0.917 and 0.935, respectively.Using the established RT-qPCR-D method, the peak of melting curve of Bmi-1, UBC and GAPDH mRNAs were single, the detection limit was up to 1.25μl.The levels of serum circulating Bmi-1 mRNA detected by RT-qPCR-D were 0.138 ( 0.078-0.228 ) in colorectal cancer stage Ⅰ patients, 0.163(0.067 -0.287) instage Ⅱ patients, 0.217(0.072-0.267) instage Ⅲpatients, 0.273(0.139 -0.419) in stage Ⅳ patients and 0.021(0.008 -0.029) in health controls, a significant difference was found among groups ( H =89.5, P <0.001 ).The levels of serum circulating Bmi-1 mRNA in each stage colorectal cancer were all significantly higher than that in control group(U=58.0, 287, 246, 72.5,all P<0.001).The levels in Ⅳstage patients were significantly higher than those in other stages patients (U=247, 590, 540,P=0.008, 0.020, 0.035), while no significant differences among Ⅰstage,Ⅱstage and Ⅲstage patients(U=633, 514, 1170,all P>0.05).ROC curve analysis showed area under the ROC curve ( AUC) for serum circulating Bmi-1 mRNA was 0.921(95%CI=0.876-0.953), which was significantly superior to the AUC of CEA (0.745, 95%CI=0.680-0.802, Z=4.697, P<0.001 ).When cutoff value was 0.034, the diagnostic sensitivity and specificity was 89.2%(141/158) and 90.6%(48/53), while 41.8%(66/158) and 73.6%(39/53) using CEA.The AUC for combination of circulating Bmi-1 mRNA and CEA was 0.933(95%CI=0.890-0.963 ) , which was no statistical significance when compared with the AUC of circulating Bmi-1 mRNA(Z=4.697, P>0.05).Conclusions The study establishes a higher sensitive, specific for detecting serum circulating Bmi-1 mRNA. Based on this method , serum circulating Bmi-1 mRNA is found to be increased in colorectal cancer , and is superior to traditional tumor marker CEA in diagnosis of colorectal cancer, which may become a potential detection index for early detection of colorectal cancer.

Objective:To analyze the alternative splicing (AS) events of patients with thyroid carcinoma (THYC) and explore the correlation between AS events and the prognosis of THYC.Methods:The clinical data and the Percent Splice In (PSI) value of AS events of THYC were downloaded from The Cancer Gene Atlas (TCGA) database and the TCGA SpliceSeq database respectively. The occurrence of seven kinds of AS events including AA, AD, AF, AP, ME, ES and RI in THYC was investigated and the matrix of AS events and survival data was constructed. Univariate Cox regression analysis was used to screen AS events related to prognosis of THYC. To avoid over-fitting, the least absolute shrinkage and selection operator (LASSO) regression analysis was performed. Then Multivariate Cox regression analysis was used to construct prognosis model. Kaplan-Meier curve and receiver operating characteristic (ROC) curve were performed to evaluate the prognosis ability of the risk model. We also used Pearson correlation analysis to select splicing factors (SF) which were correlated with survival associated AS events. Above SF genes were enrolled to gene ontology (GO) enrichment and KEGG pathway analysis.Results:A total of 10 447 genes and 45 150 AS events in 507 THYC patients were found in the present study. Among them, ES was the main type (38.84%) and ME was the type with the least frequency (0.51%). Totally 1 842 AS events associated with prognosis of THYC patients were identified. Three AS events including USHBP1-48249-AA、CACNB1-40626-AT and BEX5-89679-AP were selected to construct the prognosis model. The risk score of 0.807 was indicated as the best cut-off value of prognosis model. The patients were divided into high-risk group (240 cases) and low-risk group (241 cases) based on the risk score. The results demonstrated that the risk model could be used as a valuable prognostic factor for THYC ( P<0.001, AUC=0.929). The SF-AS network was constructed and several SF genes, including CDK12, RBM25, DDX39B, SRRM2 and DDX46 were identified as hub genes. Conclusions:The risk model based on 3-AS events was valuable prognosis predictor of THYC. The SF-AS network provided new insight for the exploration of tumorigenesis and development of THYC.

Tight-junction (TJ) is a complex supramolecular entity composed of complete membrane proteins, membranes and soluble cytoplasmic proteins, which is distributed in almost all barrier structures in the body. It can maintain the polarity of epithelial cells, close the intercellular space and prevent the overflow of materials in the epithelial space, and is a highly dynamic signaling entity. Occludin is one of the most representative members of TJ proteins, mainly responsible for sealing intercellular connections, maintaining intercellular permeability, and participating in maintaining the integrity of vascular endothelium. The integrity of occludin is related to the integrity of TJ, and the function of occludin is often associated with the barrier properties of various tissues, and the abnormal expression of occludin is related to the occurrence and development of various diseases. Occludin contains abundant Ser and Thr residues and has multiple phosphorylation sites. Phosphorylation is necessary for the combination of occludin and TJ, which can regulate the location of occludin, regulate the expression of occludin, and enhance the permeability and barrier function of TJ. Therefore, phosphorylation regulation is a mechanism that cannot be ignored in the regulation of occludin function. Occludin also interacts with many other proteins, such as co-forming the cytoskeleton with ZO-1, and is regulated by a variety of transcription factors. Studies have confirmed that in pathological conditions, a variety of signaling pathways can disrupt the integrity of cell barrier by regulating the expression and distribution of occludin. Myosin light chain kinase (MLCK) signal transduction pathway is one of the important ways to regulate the structure and function of TJ. It influences the expression of occludin by altering the cytoskeleton. MLCK mainly uses the phosphorylation of myosin light chain (MLC) as a medium to promote actin contraction, secondary decomposition of tightly binding proteins, resulting in increased or changed cellular barrier permeability, and increased MLC phosphorylation is also a biochemical marker of actomyosin contraction. Activation of MLCK causes Thr18 and Ser19 phosphorylation of MLC, which promotes the assembly of myosin II into myosin fibers and activates the hydrolysis of ATP, which relaxes the intercellular connections and reduces the ability of upper cortex to resist external invaders. Protein kinase C (PKC) plays an important role in the regulation of tightly connected signaling molecules, affecting the dynamic changes of paracellular permeability. PKC pathway is a key link in many cell signal transduction pathways, which influences all aspects of cell activities by catalyzing Ser/Thr residues phosphorylation of membrane proteins and many enzyme proteins. After PKC activation, it can regulate cellular barrier function by phosphorylating occludin and inducing its redistribution, and directly affect TJ action. Specific PKC subunits such as PKCα, PKCδ and PKCγ are activated and act on occludin molecules to promote their phosphorylation and cause the increase of TEER. The increase of TEER helps to regulate intercellular TJ and enhance the tightness of intercellular connections. Mitogen-activated protein kinases (MAPK) are usually activated by inflammatory factors, during which different signal transduction pathway subfamilies are formed to regulate occludin expression and affect tight junction and mucosal barrier functional integrity. Meanwhile, occludin is easily affected by various factors (such as cytokines and flora toxins), and abnormal expression of occludin will lead to structural damage of TJ and further damage of the intercellular barrier. Therefore, this paper summarizes the molecular structure and physiological function of occludin, and further summarizes its related signal regulation pathways and influencing factors, in order to provide theoretical support for maintaining the integrity of barrier function of occludin.