With the development of proteomic technologies, large numbers of putative biomarkers have been screened out but few of them are put into the clinical application.Recently, MRM-MS based approaches and strategies have aroused extensive concern in domestic and overseas clinical laboratory community because of their cost-efficiency, time-saving, high analytical sensitivity and accuracy.MRM-MS based approaches, especially SISCAPA, may gradually become a key technology for the protein biomarker research in the near future.In this review, the bottleneck existing in the field of protein biomarkers research, and features as well as applications of MRM-MS are discussed and the future trend of research is prospected.

Objective:To apply the strategy of proteomics with two dimensional gel electrophoresis(2DE) in esophageal cancer.Methods:the protein of esophageal cancer tissues and its adjacent normal tissue were separated by 2DE, the gels were analyzed with software ImageMaster4.01 to find out the spots different between cancer and control, and the proteins in the spots were identified by matrix associated laser desorption/ionization time of flight(MALDI TOF) mass spectrometry.Results:a new reasonable strategy of study for proteomics on esophageal cancer was raised and candidate cancer marker of two pyruvate kinase isozyme were found under such strategy.Conclusion:Under reasonable strategy, the analysis of proteomics with 2DE on human tissue is a useful method for discovering valuable cancer marker candidates. Pyruvate kinase isozyme M2/M1 which were found in cancer tussues will be very important in diagnosis and treatment of esophageal cancer.

Objective To identify the different proteins of pancreatic cancer with and without diabetes mellitus (DM) ,so as to provide some clues for understanding the molecular mechanisms of DM in the development .of pancreatic cancer. Methods Samples were divided into pancreatic cancer with or without DM and their normal adjacent tissue with 5 each. The protein expression profiles were analyzed using 2-DE in combination with MALDI-TOF/TOF MS/MS. Western blot was used to verify the expression of a candidate protein CapG. Results Seven proteins were obviously up-regulated in pancreatic cancer with DM(P＜0.05). These proteins were involved in cell motility, metabolism, oxidative stress, etc. Additionally, CapG implicated a functional contribution of cancer invasion and metastasis. Conclusion The results also provide evidence to support the hypothesis that DM plays an important role in the development of pancreatic cancer.

Objective:To analyze the causes of postoperative stricture of biliary-enteric anastomotic for congenital choledochal cysts.Methods:These 28 patients underwent salvage operation on an average 15 years (0.2-25 years) after initial surgeries at the Department of Hepatobiliary Surgery, Hunan Provincial People's Hospital from Jan 2014 to Jun 2018.Results:In 26 patients the biliary-enteric anastomotic stenosis was benign, and in 2 the stricture was caused by cancerration. In 26 cases the Roux-en-Y hepaticojejunostomy was redone,among them 8 cases underwent concurrent hepatectomy for a better exposure of the intrahepatic bile duct. In 2 cases the anastomotic stenosis was found to be caused by canceration with extensive intraabdominal metastasis ,an external drainage was adopted. There were no inhospital deaths, and no serious complications. The postoperative follow-up time was 6-67 months. Two cancerated patients died within half a year, and the remaining patients had no long-term complications.Conclusions:Biliary-enteric anastomotic stenosis is one of the serious complications in postoperative patients for congenital choledochal cysts. Hence a wide, tension free biliary-enteric anastomosis performed by a experienced hand is necessary.