AIM: To investigate the correlation between ezrin expression and invasive phenotype formation in malignantly transformed esophageal epithelial cells.
METHODS: The experimental cell line employed in the present study w...AIM: To investigate the correlation between ezrin expression and invasive phenotype formation in malignantly transformed esophageal epithelial cells.
METHODS: The experimental cell line employed in the present study was originated form the progressive induction of a human embryonic esophageal epithelial cell line (SHEE)by the E6E7 genes of human papillomavirus (HPV) type 18.The cells at the 35th passage after induction called SHEEIMM were in a state of immortalized phase and used as the control,while that of the 85th passage denominated as SHEEMT represented the status of cells that were malignantly transformed. The expression changes of ezrin and its mRNA in both cell passages were respectively analyzed by RT-PCR and Western blot. Invasive phenotype was assessed in vivo by inoculating these cells into the severe combined immunodeficient (SCID) mice via subcutaneous and intraperitoneal injection, and in vitro by inoculating them on the surface of the amnion membranes, which then was determined by light microscopy and scanning electron microscopy.
RESULTS: Upregulated expression of ezrin protein and its mRNA was observed in SHEEMT compared with that in SHEEIMM cells. The SHEEMT cells inoculated in SCID mice were observed forming tumor masses in both visceral organs and soft tissues in a period of 40 days with a special propensity to invading mesentery and pancreas, but did not exhibit hepatic metastases. Pathologically, these tumor cells harboring larger nucleus, nucleolus and less cytoplasm could infiltrate and destroy adjacent tissues. In the in vitro study,the inoculated SHEEMT cells could grow in cluster on the amniotic epithelial surface and intrude into the amniotic stroma. In contrast, unrestricted growth and invasiveness were not found in SHEEIMM cells in both in vivo and in vitroexperiment.
CONCLUSION: The upregulated ezrin expression is one of the important factors that are possibly associated with the invasive phenotype formation in malignantly transformed esophageal epithelial cells.展开更多
AIM: To systematically study the expression of carbonic anhydrase (CA) isowmes Ⅸ and Ⅻ in gastric tumors.METHODS: We analyzed a representative series of specimens from non-neoplastic gastric mucosa and from various ...AIM: To systematically study the expression of carbonic anhydrase (CA) isowmes Ⅸ and Ⅻ in gastric tumors.METHODS: We analyzed a representative series of specimens from non-neoplastic gastric mucosa and from various dysplastic and neoplastic gastric lesions for the expression of CA IX and XII. Immunohistochemical staining was performed using isozyme-specific antibodies and biotinstreptavidin complex method.RESULTS: CA IX was highly expressed in the normal gastric mucosa and remained positive in many gastric tumors. In adenomas, CA IX expression significantly decreased towards the high grade dysplasia. However, the expression resumed back to the normal level in well differentiated adenocarcinomas,while it again declined in carcinomas with less differentiation.In comparison, CA Ⅻ showed no or weak immunoreaction in the normal gastric mucosa and was slightly increased in tumors.CONCLUSION: These results demonstrate that CA Ⅸexpression is sustained in several types of gastric tumors.The variations observed in the CA Ⅸ levels support the concept that gastric adenomas and carcinomas are distinct entities and do not represent progressive steps of a single pathway.展开更多
Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of...Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of HBV in E.coli.Methods.The genes of HBcAg (1-144)and preS1(1-42)were amplified and fused by PCR.This fused gene was inserted in the prokaryotic expression vector pET-11d and expre ssed in E.coli.Results.It was showed by SDS-PAGE that the pr otein molecular weight of the coexpr ession product was about 20kD,20%of all bacteria prote in.The monoclonal antibodies again st core and preS1antibody could re-act with this fused protein by Western-blot technique respectively.The fused gene was verified by sequencin g.Under the immune electron microscop y,this fused protein is typical particles of HBcAg but in an aggregated form.Conclusion.The results might aid for studying T c ell immunotherapeutic vaccine for c hronic hepatitis B.展开更多
Cases (n=44) with squamous cell cervical cancer (SCCA) and age-matched healthy controls (n=176) were analyzed. Significant difference due to human papillomavirus (HPV) infection, ages at the first marriage, ages at th...Cases (n=44) with squamous cell cervical cancer (SCCA) and age-matched healthy controls (n=176) were analyzed. Significant difference due to human papillomavirus (HPV) infection, ages at the first marriage, ages at the first sexual intercourse, ages at the first birth given to baby, number of gravidities, number of deliveries, Body Mass Index (BMI), education level of women and their husbands (p<0.05) was observed. According to multivariate logistic regression analysis, four factors have entered the model (p<0.05), including HPV infection [OR (odds ratio)=26.13, 95%CI (confidence interval)=9.40?72.60], education level of women (OR=0.41, 95%CI=0.21?0.79), education level of spouses (OR=0.45, 95%CI=0.22?0.94), BMI (OR=0.73, 95%CI=0.57?0.93). Moreover, HPV infection is relative to education level of women (r=?0.14), and their spouses (r=?0.21), age at the first marriage (r=?0.20), age at the first birth given to baby (r=?0.20) and BMI (r=?0.15).展开更多
An E1B-defective adenovirus named r1/Ad was constructed by homologous recombination. The construction, selection and propagation of recombinant virus was done in the human embryonic kidney 293 cells (HEK293). Thein vi...An E1B-defective adenovirus named r1/Ad was constructed by homologous recombination. The construction, selection and propagation of recombinant virus was done in the human embryonic kidney 293 cells (HEK293). Thein vitro study demonstrated that the recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as the human glioblastoma tumor cells (U251) and human bladder tumer cells (EJ) but not in the normal cells with functional p53 such as the human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated that the U251 cells were more sensitive to the infection of r1/Ad than that of EJ cells under identical conditions. In this paper, it was found that r1/Ad could be very useful in studying thein vitro selective replication of E1B-defective adenovirus. This may help to determine the safety of using any E1B-defective adenoviruses in cancer gene therapy. Key words E1B-defective adenovirus - cytopathic effect - cancer gene therapy CLC number Q78 Foundation item: Supported by the National Natural Science Foundation of China (3988003)Biography: Xue Feng (1978-), male, Master, research direction: cancer gene therapy by recombinant virus.展开更多
The recombinant adeno-associated viral vector with human coagulation Factor Ⅸ minigene which wasregulated by CMV promoter was constructed. Largequantity of recombinant adeno-associated viral particles (rAAV/ hFⅨ) wa...The recombinant adeno-associated viral vector with human coagulation Factor Ⅸ minigene which wasregulated by CMV promoter was constructed. Largequantity of recombinant adeno-associated viral particles (rAAV/ hFⅨ) was prepared by the HSV/AAV hybrid helper virus method. Southern dot blot assay and QC-PCR indicated that the titer of the virus was 3.6×1012 v.g./mL. It demonstrated that this method can effectively overcome the hurdles of mass production of AAV vector. Followed by anintramuscular injection of viral vectors (7.5×1011 v.g./mouse) in the quadriceps femoris, an elevation of human Factor Ⅸexpression in the plasma of hemophilia B mice was detected (387 ng/mL) and persisted more than 12 weeks. The level of anti-virus antibody in plasma aligned with the Factor Ⅸexpression curve. The QC-PCR method is easier and moreaccurate than traditional dot hybridization fordetermination of the titer of recombinant adeno-associated virus. Moreover, there are no HSV particles existing inproduced AAV assayed by RT-PCR. AAV is the only virus that has been amplified from AAV-injected muscle by PCR.展开更多
The Fourth China National Congress on Medical Virology was held from October 26 to 31, 1993 in Xiamen, Fujian Province. A total of 217 virology workers participated in the congress, which received 312 papers, 82 of wh...The Fourth China National Congress on Medical Virology was held from October 26 to 31, 1993 in Xiamen, Fujian Province. A total of 217 virology workers participated in the congress, which received 312 papers, 82 of which were selected for oral presentations. The plenary session was held on October 27, 1993. Four invited lectures were delivered first, followed by five presentations. The workshops were divided into 3 groups on October 28 and the morning of October 29.The first group included hemorrhagic fever with renal syndrome virus and arboviruses. The second group included enteric viruses, rabies virus and other viruses. The third group incluffed hepatitis viruses and herpes viruses.展开更多
The three-dimensional structure of Periplaneta fuliginosa densovirus (pfDNV) is determined at 2.3 nm reso-lution using the techniques of cryo-electron microscopy and image reconstruction. The pfDNV contains five struc...The three-dimensional structure of Periplaneta fuliginosa densovirus (pfDNV) is determined at 2.3 nm reso-lution using the techniques of cryo-electron microscopy and image reconstruction. The pfDNV contains five structural proteins and 60 protein subunits arranged on a T = 1 icosa-hedral shell with a relatively smooth surface. Its recon-struction reveals its distinct capsid structure from those ob-served in CPV and GmDNV. As in GmDNV, spike-like protrusions are not present in pfDNV at the threefold axes; while two small thorn-like protrusions are identified there. However, different from CPV and GmDNV, cylindrical channels along the fivefold axes are closed in pfDNV; while a small thorn-like protrusions, which have not been reported in other parvovirus, are observed there in pfDNV although their function is yet to be investigated. The pfDNV has dim-ple-like depressions at the icosahedral twofold axes; but has no canyon-like regions encircling the fivefold axes. The ico-sahedrally well-ordered nucleic acid has also been observed in pfDNV, suggesting that the protein and nucleic acid probably form closed interaction.展开更多
基金the National Natural Science Foundation of China (No.39830380,39900069)Research and Development Foundation of Shantou University(L00012)the Chinese National Human Genome Center,Beijing
文摘AIM: To investigate the correlation between ezrin expression and invasive phenotype formation in malignantly transformed esophageal epithelial cells.
METHODS: The experimental cell line employed in the present study was originated form the progressive induction of a human embryonic esophageal epithelial cell line (SHEE)by the E6E7 genes of human papillomavirus (HPV) type 18.The cells at the 35th passage after induction called SHEEIMM were in a state of immortalized phase and used as the control,while that of the 85th passage denominated as SHEEMT represented the status of cells that were malignantly transformed. The expression changes of ezrin and its mRNA in both cell passages were respectively analyzed by RT-PCR and Western blot. Invasive phenotype was assessed in vivo by inoculating these cells into the severe combined immunodeficient (SCID) mice via subcutaneous and intraperitoneal injection, and in vitro by inoculating them on the surface of the amnion membranes, which then was determined by light microscopy and scanning electron microscopy.
RESULTS: Upregulated expression of ezrin protein and its mRNA was observed in SHEEMT compared with that in SHEEIMM cells. The SHEEMT cells inoculated in SCID mice were observed forming tumor masses in both visceral organs and soft tissues in a period of 40 days with a special propensity to invading mesentery and pancreas, but did not exhibit hepatic metastases. Pathologically, these tumor cells harboring larger nucleus, nucleolus and less cytoplasm could infiltrate and destroy adjacent tissues. In the in vitro study,the inoculated SHEEMT cells could grow in cluster on the amniotic epithelial surface and intrude into the amniotic stroma. In contrast, unrestricted growth and invasiveness were not found in SHEEIMM cells in both in vivo and in vitroexperiment.
CONCLUSION: The upregulated ezrin expression is one of the important factors that are possibly associated with the invasive phenotype formation in malignantly transformed esophageal epithelial cells.
基金Sigrid Juselius FoundationFinnish Cultural Foundation and Finnish Dental Society+2 种基金the National Institutes of Health(DK40163,GM34182,and GM53405)Slovak Grant Agency(2/7152/20)Bayer Corporation
文摘AIM: To systematically study the expression of carbonic anhydrase (CA) isowmes Ⅸ and Ⅻ in gastric tumors.METHODS: We analyzed a representative series of specimens from non-neoplastic gastric mucosa and from various dysplastic and neoplastic gastric lesions for the expression of CA IX and XII. Immunohistochemical staining was performed using isozyme-specific antibodies and biotinstreptavidin complex method.RESULTS: CA IX was highly expressed in the normal gastric mucosa and remained positive in many gastric tumors. In adenomas, CA IX expression significantly decreased towards the high grade dysplasia. However, the expression resumed back to the normal level in well differentiated adenocarcinomas,while it again declined in carcinomas with less differentiation.In comparison, CA Ⅻ showed no or weak immunoreaction in the normal gastric mucosa and was slightly increased in tumors.CONCLUSION: These results demonstrate that CA Ⅸexpression is sustained in several types of gastric tumors.The variations observed in the CA Ⅸ levels support the concept that gastric adenomas and carcinomas are distinct entities and do not represent progressive steps of a single pathway.
文摘Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of HBV in E.coli.Methods.The genes of HBcAg (1-144)and preS1(1-42)were amplified and fused by PCR.This fused gene was inserted in the prokaryotic expression vector pET-11d and expre ssed in E.coli.Results.It was showed by SDS-PAGE that the pr otein molecular weight of the coexpr ession product was about 20kD,20%of all bacteria prote in.The monoclonal antibodies again st core and preS1antibody could re-act with this fused protein by Western-blot technique respectively.The fused gene was verified by sequencin g.Under the immune electron microscop y,this fused protein is typical particles of HBcAg but in an aggregated form.Conclusion.The results might aid for studying T c ell immunotherapeutic vaccine for c hronic hepatitis B.
文摘Cases (n=44) with squamous cell cervical cancer (SCCA) and age-matched healthy controls (n=176) were analyzed. Significant difference due to human papillomavirus (HPV) infection, ages at the first marriage, ages at the first sexual intercourse, ages at the first birth given to baby, number of gravidities, number of deliveries, Body Mass Index (BMI), education level of women and their husbands (p<0.05) was observed. According to multivariate logistic regression analysis, four factors have entered the model (p<0.05), including HPV infection [OR (odds ratio)=26.13, 95%CI (confidence interval)=9.40?72.60], education level of women (OR=0.41, 95%CI=0.21?0.79), education level of spouses (OR=0.45, 95%CI=0.22?0.94), BMI (OR=0.73, 95%CI=0.57?0.93). Moreover, HPV infection is relative to education level of women (r=?0.14), and their spouses (r=?0.21), age at the first marriage (r=?0.20), age at the first birth given to baby (r=?0.20) and BMI (r=?0.15).
文摘An E1B-defective adenovirus named r1/Ad was constructed by homologous recombination. The construction, selection and propagation of recombinant virus was done in the human embryonic kidney 293 cells (HEK293). Thein vitro study demonstrated that the recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as the human glioblastoma tumor cells (U251) and human bladder tumer cells (EJ) but not in the normal cells with functional p53 such as the human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated that the U251 cells were more sensitive to the infection of r1/Ad than that of EJ cells under identical conditions. In this paper, it was found that r1/Ad could be very useful in studying thein vitro selective replication of E1B-defective adenovirus. This may help to determine the safety of using any E1B-defective adenoviruses in cancer gene therapy. Key words E1B-defective adenovirus - cytopathic effect - cancer gene therapy CLC number Q78 Foundation item: Supported by the National Natural Science Foundation of China (3988003)Biography: Xue Feng (1978-), male, Master, research direction: cancer gene therapy by recombinant virus.
基金supported by the National Natural Science Foundation of China(Grant No.30100102).
文摘The recombinant adeno-associated viral vector with human coagulation Factor Ⅸ minigene which wasregulated by CMV promoter was constructed. Largequantity of recombinant adeno-associated viral particles (rAAV/ hFⅨ) was prepared by the HSV/AAV hybrid helper virus method. Southern dot blot assay and QC-PCR indicated that the titer of the virus was 3.6×1012 v.g./mL. It demonstrated that this method can effectively overcome the hurdles of mass production of AAV vector. Followed by anintramuscular injection of viral vectors (7.5×1011 v.g./mouse) in the quadriceps femoris, an elevation of human Factor Ⅸexpression in the plasma of hemophilia B mice was detected (387 ng/mL) and persisted more than 12 weeks. The level of anti-virus antibody in plasma aligned with the Factor Ⅸexpression curve. The QC-PCR method is easier and moreaccurate than traditional dot hybridization fordetermination of the titer of recombinant adeno-associated virus. Moreover, there are no HSV particles existing inproduced AAV assayed by RT-PCR. AAV is the only virus that has been amplified from AAV-injected muscle by PCR.
文摘The Fourth China National Congress on Medical Virology was held from October 26 to 31, 1993 in Xiamen, Fujian Province. A total of 217 virology workers participated in the congress, which received 312 papers, 82 of which were selected for oral presentations. The plenary session was held on October 27, 1993. Four invited lectures were delivered first, followed by five presentations. The workshops were divided into 3 groups on October 28 and the morning of October 29.The first group included hemorrhagic fever with renal syndrome virus and arboviruses. The second group included enteric viruses, rabies virus and other viruses. The third group incluffed hepatitis viruses and herpes viruses.
基金supporled by the National Natural Science Foundation of China(Grant No.39470032)
文摘The three-dimensional structure of Periplaneta fuliginosa densovirus (pfDNV) is determined at 2.3 nm reso-lution using the techniques of cryo-electron microscopy and image reconstruction. The pfDNV contains five structural proteins and 60 protein subunits arranged on a T = 1 icosa-hedral shell with a relatively smooth surface. Its recon-struction reveals its distinct capsid structure from those ob-served in CPV and GmDNV. As in GmDNV, spike-like protrusions are not present in pfDNV at the threefold axes; while two small thorn-like protrusions are identified there. However, different from CPV and GmDNV, cylindrical channels along the fivefold axes are closed in pfDNV; while a small thorn-like protrusions, which have not been reported in other parvovirus, are observed there in pfDNV although their function is yet to be investigated. The pfDNV has dim-ple-like depressions at the icosahedral twofold axes; but has no canyon-like regions encircling the fivefold axes. The ico-sahedrally well-ordered nucleic acid has also been observed in pfDNV, suggesting that the protein and nucleic acid probably form closed interaction.
