Objective To construct a new recombinant immunotoxin expression vector fused with a murine interleukin18(IL18) gene and a truncated pseudomonas exotoxin (PE38) gene, and examine the expression of IL-18-PE38 fusion protein in Escherichia coli (E. coli). Method Murine IL-18 (mIL-18) cDNA was cloned from murine liver tissue through reverse transcription-polymerase chain reaction (RT-PCR). The mIL-18 cDNA was ligased with a PE38 gene carried by PRKL expression vector through T4 DNA ligase and constructed into fusion protein expression plasmid PRKL-IL18-PE38. The recombinant vector was identified by restriction endonucleases digestion, PCR and DNA sequencing. After transformed into E.coli BL21 and induced by IPTG, the expressed product was obtained and the molecular weight and specificity were determined by SDS-PAGE and Western-blotting. Result The new recombinant immunotoxin expression vector was constructed successfully. DNA sequencing revealed that the mIL-18 and PE38 gene were consistent with NCBI Gene Bank. The IL-18-PE38 fusion protein was expressed in E.coli BL21, and Western-blotting analysis indicated that the molecular weight of the expression product is about 56 kDa, and could react with the specific antibody against mIL-18. Conclusion IL-18-PE38 recombinant immunotoxin expression vector will provide the basis for study on the targeted cytotoxic activity to Th1 cells and may have some potential value in the treatment of Th1 cell-mediated autoimmune diseases.

Objective To approach the expression and significance of CD30 in Hodgkin lymphoma cells which were transfected with mtr Ⅱ gene. Methods The CD30 expression of Hodgkin lymphoma cells in those transfected with mtr Ⅱ gene was analysed by immunohistochemical technique respectively. Results The CD30 expression rate of Hodgkin lymphoma cells which were transfected with mtr Ⅱ gene was higher than that of Hodgkin lymphoma cells which weren't transfected with mtrⅡ gene (P ＜0.01). Conclusion The mtr Ⅱ gene may be related to the malignant transformation of Hodgkin lymphoma.

Objective To investigate the clinical features, effective treatment, survival and prognostic factors of second primary tongue squamous cell carcinoma (SPTSCC) after nasopharyngeal carcinoma (NPC) radiotherapy. Methods The clinical data of 35 cases with SPTSCC after NPC radiotherapy were analyzed retrospectively. Kaplan-Meier method, Log-Rank test and COX proportional hazard mode was performed for statistical analysis. Results 3-year and 5-year overall survival rates were 55 % and 47 %, respectively, lymph node metastasis rate was 5.71 %. Univariate analysis indicated that gender (χ2 = 8.89, P = 0.00), T classification (χ2= 5.58, P= 0.02), clinical stage (χ2 = 8.51, P= 0.04) and treatment methods (χ2 = 29.37, P = 0.00) were important factors of prognosis. Multivariate analysis showed that treatment methods (P = 0.00) and T classification (P = 0.03) were independent prognostic factors. Operative treatment group had better prognosis than the non-operative treatment group, the difference was statistically significant (P <0.05), male patients in the risk of SPTSCC was higher than the female patients, and the incidence of SPTSCC was increased along with extension of the time after NPC radiotherapy. Conclusion The rate of the lymph node metastasis is lower for SPTSCC after NPC radiotherapy and treatment patterns and T stage are independent prognostic factors. Long-term follow-up after NPC radiotherapy is necessary to the early diagnosis of SPTSCC, so that to give surgery or combined therapy with surgery in order to achieve a good effect.

Objective To approach the proliferative activities of Hodgkin's disease cells were transfected with mtrⅡ gene.Methods The ki67 and PCNA expression of Hodgkin's disease cells and those which were transfected with mtrⅡ gene by immunohistochemical technique respectively.Results The expression rate of ki67 and PCNA in Hodgkin's disease cells which were transfected with mtrⅡ gene was higher than that weren't transfected(P

OBJECTIVE: To identify long-term outcomes and safety of transplantation of autologous peripheral blood stem cells (PBSC) for treating dilated cardiomyopathy.METHODS: A total of 38 cases with dilated cardiomyopathy received treatment at the Department of Cardiology, Guangdong General Hospital of Chinese People's Armed Police Forces, were selected, including 26 males and 12 females, aged 42-72 years, mean aged 56 years. Based on given standard therapy, 38 patients divided randomly into the transplantation group (n=20) and the control group (n=18). Patients in the transplantation group were received recombinant human granulocyte colony-stimulating factor (rhG-CSF) 300 ug/d once per day for 5 days to mobilize stem cells. At day 6, PBSC were collected with blood-cells separator and were transplanted through intracoroary way. The routine medication was performed in the control group. Blood routine test, hepatic function, renal function, glucose, triglyeride (TG), cholesterol, low density cholesterol (C-LDL), high density cholest- erol (C-HDL), uric acid (UA), creatine kinase (CK), isoenzyme of creatine kinase (CK-MB) and high sensitive C-reactive protein (hsCRP) were measured before and at months 6 and 12 after transplantation. All patients also received ultrasonic echocardiography, ECG Holter monitor and six-minute-walk test before and at 12 and 24 months after the procedure. Survival rate and incidence rate of heart incidents were compared. The study end-point was death from any cause. RESULTS: All patients received a 12-24 month follow-up with mean (18±6) months. One patient in the transplantation group received mitral valve replacement. One patient of the transplantation group and 2 of the control group died due to refractory heart failure. The blood routine test and biochemical indicators of the transplantation group had no significant differences among 6 months and 12 months after transplantation compared with control and pre-transplantation (P > 0.05). Six-minute-walking distance in the transplantation group significantly increased at 12 months after transplantation than pre-transplantation level, which was also higher than that of control patients (P < 0.05). The left ventricular ejection fraction (LVEF) was increased (P < 0.01). The left ventricular diastolic diameter (LVDd) decreased significantly in the transplantation group (P < 0.01). In the control group, improvement in LVEF and LVDd were observed, but there was no significant difference (P > 0.05). After 24 months of follow-up, the above-mentioned indexes had not improved in the transplantation group without significant differences. No malignant arrhythmias and severe side effects could be observed around transplantation and during 24 months follow-up. Survival was similar between the two groups during 24 months follow. CONCLUSION: Transplantation of mobilized autologous PBSC might be a safe and effective method for the treatment of dilated cardiomyopathy, which may improve the ventricular systolic function in a short-term, however, the long-term effects still uncertain.

ObjectiveTo detect BAG1 expressions in digestive tract cancer by tissue microarray and to evaluate its clinical significance.MethodsTissue microarray of digestive tract cancer and normal tissues were analyzed by DAKO Envision system immunohistochemical staining for apoptosis related gene BAG-1 expression.ResultsThe positive rate of BAG-1 expression among esophagus cancer,gastric cancer and rectal cancer were higher than that of normal tissues respectively(P<0.01).ConclusionThere is an overexpression of BAG-1 in digestive tract cancer,which suggest that apoptosis related gene BAG-1 may be related to these cancer.

Objective To investigate the proliferative activities and significance of Hodgkin's disease cells which were transfected with mtr Ⅱ gene. Methods The ki67 and proliferating cell nuclear antigen (PCNA) expression in Hodgkin's disease cells transfected with mtr Ⅱ gene were determined with immunohistochemistry. Results The ki67 and PCNA expression in cells transfected with mtr Ⅱ gene was more than that of cells without mtrⅡ gene transfected (P<0.05). Conclusion The proliferative activities of Hodgkin's disease cells transfected with mtr Ⅱ gene increased obviously, which may be related to the progress of Hodgkin's disease.

OBJECTIVETo study the apoptotic effects of influenza A virus on the Raji cell line.

METHODSCultured Raji cells were infected with influenza A virus at a multiplicity of infection (m.o.i) of 20 and the effects of apoptosis were detected at different time points post infection using the following methods: electron microscope, DNA agarose gel electrophoresis, PI stained flow cytometry (FCM) and Annexin-V FITC/PI stained FCM.

RESULTSRaji cells infected with influenza A virus showed changes of morphology apoptosis, DNA agarose electrophoresis also demonstrated a ladder-like pattern of DNA fragments in a time-dependent manner. PI stained FCM showed "apoptosis peak" and FITC/PI stained FCM showed apoptotic cells. Quantitative analysis indicated that the percentage of apoptotic Raji cells increased after infection, and cycloheximide (CHX), an eukaryotic transcription inhibitor, could effectively inhibit the apoptotic effects of influenza A virus in vitro.

CONCLUSIONSInfluenza A virus can induce apoptosis in Raji cell line suggesting that it may lead to a potential method for tumor therapy.

Humans ; Male ; Middle Aged ; Pedigree ; Porokeratosis ; genetics

Adult ; Female ; Humans ; Male ; Pedigree ; Tuberous Sclerosis ; diagnosis