Proteomic-based analysis for identification of potential serum biomarkers in gallbladder cancer.

Gallbladder cancer is the most common malignant tumor of the biliary tract. Early diagnosis of gallbladder cancer is difficult because of the latent onset and lack of good biomarkers. To identify new biomarkers that improve the early diagnosis and/or serve as possible therapeutic targets in gallbladder cancer is essential. In the present study, serum proteins were separated by two-dimensional gel electrophoresis (2-DE) in 3 patients with gallbladder cancer and 3 healthy volunteers. The differentially expressed spots were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Western blotting and immunohistochemistry were performed to verify the expression of certain candidate proteins. Protein expression and clinical correlation was evaluated. We found that 64 protein spots were significantly changed in gallbladder cancer. Twenty-four proteins including S100A10, haptoglobin, cystatin-B, profilin-1 and superoxide dismutase were successfully identified. Among these proteins, S100A10 and haptoglobin were validated using Western blotting. Immunohistochemically, the expression of S100A10 and haptoglobin proteins was found to be higher in gallbladder cancer tissues compared to that in gallbladder adenoma, liver cholangiocarcinoma and cholecystitis tissue. Patients with high expression of S100A10 and haptoglobin were linked to late stage disease and poor clinical prognosis. Our data suggest that combined comparative proteomic analysis by 2-DE and MALDI-TOF-MS is an effective method for identifying differentially expressed proteins in serum samples. These proteomic approaches could be used for identifying new serum biomarkers in gallbladder cancer. S100A10, haptoglobin and other identified proteins may be potential molecular targets for early gallbladder cancer diagnostics and therapeutic applications.

[Surgery for T4 hypopharyngeal cancer and reconstruction after hypopharyngo-oesphagectomy].

OBJECTIVE: To explore the surgical treatment of hypopharyngeal and cervical esophageal cancers and the ways of reconstruction after hypopharyngo-oesphagectomy, and to evaluate their efficacy. METHODS: Twenty five patients with cancer of the laryngopharynx and cervical esophagus treated in our department between 1995 and 2007 were included in this study. Their clinical data were restrspectively analyzed. Among them, 17 cases had the tumor originated from the pyriform sinus, 3 of the posterior pharyngeal wall and 5 of the postcricoid region. Acording to the 2002 UICC criteria, all the tumors were stage T4, including 9 patients with cN0, 11 with cN1, and 5 with cN2 disease. The pharyngoesophageal defect reconstruction methods were as following: pharyngogastric anastomosis in 7 patients, free jejunal transplantion in 4, laryngotracheal flap in 8, and pectoralis major musculocutaneous flap in 6 patients. All patients were treated with modified and/or selective neck dissection. Among them, 8 cases received pre-operation radiotherapy, 17 received post-operative auxiliary radiotherapy. RESULTS: There was no operation death case in this group. All patients were followed up for 3 to 5 years. Three patients died in the first year. According to Kaplan-Meier analysis, the 1-year survival rate was 88.0%, 3-year survival rate was 48.0%, and 5-year survival rate was 28.0%. CONCLUSIONS: The use of primary repair of the defects of laryngopharynx and cervical esophagus expands the operative indication for cancers of the laryngopharynx and cervical esophagus, improves the survival rate and life quality of the patients. Regarding the repair method of choice, site of the tumor and size of the defect are the most important factors regarding choice of reconstruction method, while the patients' age and general condiction should also be considered to minimize the complications as more as possible.

[Immunological killing effect of recombicant adenovirus vector rAD-mTERT-m4-1BBL on mouse hepatoma cell line Hepa1-6 cells co-cultured with T lymphocytes].

OBJECTIVE: To study the immunological suppressing effect of recombinant adenovirus vector rAD-mTERT promotor-m4-1BBL (rAD-mTERT) on mouse hepatoma cell line Hepa1-6 cells in co-culture with T lymphocytes. METHODS: Adding recombinant adenovirus rAD, rAD-CMV-m4-1BBL (rAD-CMV) and rAD-mTERT to Hepa1-6 and L929 cells, respectively, to observe the effect of these adenoviruses on growth and apoptosis of these cells in co-culture with T lymphocytes. RESULTS: Adding adenovirus significantly suppressed the growth and slightly increased apoptosis of the two types of cells (P < 0.05). rAD-mTERT promotor-m4-1BBL showed only pro-apoptotic effect on Hepa1-6 cells. When co-cultured with T lymphocytes, rAD-CMV-m4-1BBL showed promoting effect on apoptosis of the cells. Compared with that of T cells pre-co-culture, CD4(+) and CD8(+) T cells were proliferated, and the ratio of CD4/CD8 was significantly reduced (from 1.27 to 1.08). CONCLUSION: Adding the recombinant adenoviruses only suppresses the cell growth, but not promotes their apoptosis. In co-culture with T lymphocytes, recombinant adenovirus vector rAD-mTERT promotor-m4-1BBL can targetingly suppress the growth and induce apoptosis of Hepa1-6 cells. The apoptosis is induced through the immunological killing effect of T lymphocytes.

[The application of epiglottis and cervical anterior muscle to laryngeal reconstruction].

OBJECTIVE: To evaluate the methods of reconstruction of laryngeal function after the extended laryngectomy. METHODS: The laryngeal functions were reconstructed after the extended laryngectomy performed on 22 cases with the laryngocarcinoma of T2 and T3 stages of glottic type, using epiglottis and cervical anterior muscle to reconstruct the laryngeal cavity and larynx bracket, from September 1998 to October 1999. RESULTS: Twenty cases recovered normal swallow function in 20 days post-operation as well as the entire function of larynx was recovered, and the rate of decannulation was 90.9% (20/22). One case with pharyngeal fistula recovered a month later, but the other case recovered to normal diet by operation repaired. All the patients pronounced clearly and were able to keep the characteristics of their own voice. The fibred laryngoscope examination showed that the sphincter valves were formed at the larynx atrium in all postoperative cases. The rate of 3-year survival for T2 and T3 stages was 100% (9/9) and 92.3% (12/13), respectively. CONCLUSION: The laryngeal cavity could be enlarged and the laryngeal atrium was reconstructed by employing epiglottis ifraplacement and cervical anterior muscle to reduce the tension of epiglottis and minimize the injury of the membrane as well as to overcome the shortcomings of insufficient material if mere employing epiglottis. The application of combined methods to reconstruct the laryngeal cavity is a practical way to restore the larynx function and improve the life quality.