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Objective:To analyze the factors affecting lymph node failure pattern after radiotherapy/chemotherapy in esophageal squamous cell carcinoma without initial clinical metastasis. Methods:A total of 263 patients, who were diagnosed as thoracic esopha-geal carcinoma from January 2002 to December 2009, were included in this retrospective study. Factors affecting lymph node failure pattern with general clinical data and tumor local factors were analyzed. Results:Among the 263 esophageal cancer cases, 31 (11.8%) had lymph node metastasis after treatment, including 18 cases of simple lymph node metastasis and 13 other cases of lymph node metas-tasis with esophageal and other organ metastasis or recurrence. The numbers of cases for lymph node metastasis in the upper, middle, and lower thoracic esophagus were 11 (13.3%), 13 (10.1%), and 7 (13.7%), respectively. Univariate analysis showed that recent cura-tive effect, length of tumor on X-rays, maximum tumor diameter, and tumor volume were the significant factors associated with lymph node metastasis (χ2=7.597, 9.717, 5.361, and 4.815;P=0.006, 0.002, 0.021, and 0.028). Logistic regression analysis results showed that recent curative effect and length of tumor on X-rays were independent significant factors (P=0.004 and 0.026). Conclusion:Recent cu-rative effect and length of tumor on X-rays were the significant factors associated with lymph node failure pattern after radiotherapy/chemotherapy in esophageal squamous cell carcinoma without initial clinical metastasis.
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Objective To prepare and characterize the polyclonal antibody against N protein of SARS-CoV. Methods The polyclonal antibody against SARS-CoV N was obtained from immunized rabbit with purified GST-N. The titer of the antibody was determined by indirect ELISA, and the specificity by Western blot and immunochemical staining. Results The rabbit′s antibody against SARS-CoV N was prepared successfully. The titer of antiserum against SARS-CoV N was about 1.2?10~ -5 . Western blot and immunochemical staining analysis showed that the polyclonal antibody could bind to the expressed fusion protein specifically. Conclusion The rabbit′s antibody against SARS-CoV N has been prepared successfully, and it can be a useful reagent for clinical diagnosis and further research.
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Objective To construct an expression vector pGEX-2T/N, and to express the fusion protein consisting of N protein of SARS-CoV in E. coli.Methods The N region gene of SARS-CoV was obtained by RT-PCR. The expression vector PGEX-2T/N was constructed by DNA recombination. The recombinant plasmid was transformed into E. coli BL21(DE3). The expression of the fusion protein was determined by Western blot with anti-SARS-CoV antibody positive blood sera. Results The N region gene of SARS-CoV was obtained. The fusion protein GST-N was soluble. Western blot analysis showed that the reaction of GST-N to anti-SARS-CoV sera was positive. Conclusion The pGEX-2T/N has been constructed and expressed in the form of fusion protein GST-N successfully, and the result lays the foundation for further study of SARS-CoV N protein.
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Objective To compare the cellular immune responses induced by two different vectors,eukaryotic plasmid pcDN3.1(+) and attenuated typhia live vaccine Ty21a,-associated mimic epitopes of HCV hypervariable region 1,so to find a better way for vaccine immunization.Methods The DNA sequence based on the peptides' sequence of HCV hypervariable region 1 related consensus polymimotopes was synthesize and then inserted into plasmid pcDN3.1(+) to construct recombinant plasmid pcDN3.1(+)-SP,which then was transfected into live attenuated typhia vaccine Ty21a to construct Ty21a-SP.The mice were immunized orally with Ty21a-SP and intramuscularly with pcDN3.1(+)-SP,respectively,and then sacrificed by exsanguination.The splenocytes were separated and restimulated with pooled synthesized peptides,and then collected.Flow cytometry was employed to identify CD8+IFN-?+ T cells,and non-radioactive MTS method was adopted to test T cell proliferation,and non-radioactive LDH method was used to test cytotoxic T cytolytic reaction(CTL).Results Compared with control groups,the proliferation of splenocytes was apparently enhanced,the proportion of CD8+IFN-?+ cells obviously increased,and CTL responses also significantly increased after the spenocytes of mice immunized by pcDN3.1-SP and Ty21a-SP were restimulated with synthesized peptides.And the responses mentioned above in the mice immunized by Ty21a-SP were stronger than those mice immunized by pcDN3.1-SP.Conclusion Using attenuated typhia live vaccine Ty21a as DNA vector is an effective way to induce cellular immune responses.