AIM: Based on the pathogenesis of severe acute pancreatitis and our experimental studies, to investigate the effect of dexamethasone and dextran in treatment of patients with severe acute pancreatitis.METHODS: Thirty-...AIM: Based on the pathogenesis of severe acute pancreatitis and our experimental studies, to investigate the effect of dexamethasone and dextran in treatment of patients with severe acute pancreatitis.METHODS: Thirty-two patients with severe acute pancreatitis were treated with 0.5-1 mg/kg per day dexamethasone for 3-5 d, and 500-1 000 mL/d of dextran 40 for 7 d, besides the routine therapy.RESULTS: After 4-8 h of treatment, abdominal pain began to be relieved; range of tenderness began to be localized in 27 patients. They were cured with nonsurgical treatment.Five of them were deteriorated, and treated with surgery.Four patients in this group died.CONCLUSION: Dexamethasone and dextran 40 block the pathologic process of severe acute pancreatitis through inhibition of inflammatory mediators and improvement of microcirculation disorders respectively.展开更多
AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse...AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing,the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.RESULTS: ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E.coli M15 and expressed. The expressed ODC protein was verified with Western blotting.CONCLUSION: The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.展开更多
AIM: To investigate the ornithine decarboxylase (ODC)gene expression in colorectal carcinoma, ODC mRNA was assayed by RT-PCR and ODC protein was detected by a monoclonal antibody against fusion of human colon ODC prep...AIM: To investigate the ornithine decarboxylase (ODC)gene expression in colorectal carcinoma, ODC mRNA was assayed by RT-PCR and ODC protein was detected by a monoclonal antibody against fusion of human colon ODC prepared by hybridoma technology.METHODS: Total RNA was extracted from human colorectal cancer tissues and their normal counterpart tissues. ODC mRNA levels were examined by RT-PCR.ODC genes amplified from RT-PCR were cloned into a prokaryotic vector pQE-30. The expressed proteins were purified by chromatography. Anti-ODC mAb was prepared with classical hybridoma techniques and used to determine the ODC expression in colon cancer tissues by immunohistochemical and Western blotting assay.RESULTS: A cell line, which could steadily secrete antiODC mAb, was selected through subcloning four times.Western blotting reconfirmed the mAb and ELISA showed that its subtype was IgG2a. RT-PCR showed that the ODC mRNA level increased greatly in colon cancer tissues (P＜0.01). Immunohistochemical staining showed that colorectal carcinoma cells expressed a significantly higher level of ODC than normal colorectal mucosa (98.6±1.03%vs 5.26±5%, P＜0.01).CONCLUSION: ODC gene overexpression is significantly related to human colorectal carcinoma. ODC gene expression may be a marker for the gene diagnosis and therapy of colorectal carcinoma.展开更多
文摘AIM: Based on the pathogenesis of severe acute pancreatitis and our experimental studies, to investigate the effect of dexamethasone and dextran in treatment of patients with severe acute pancreatitis.METHODS: Thirty-two patients with severe acute pancreatitis were treated with 0.5-1 mg/kg per day dexamethasone for 3-5 d, and 500-1 000 mL/d of dextran 40 for 7 d, besides the routine therapy.RESULTS: After 4-8 h of treatment, abdominal pain began to be relieved; range of tenderness began to be localized in 27 patients. They were cured with nonsurgical treatment.Five of them were deteriorated, and treated with surgery.Four patients in this group died.CONCLUSION: Dexamethasone and dextran 40 block the pathologic process of severe acute pancreatitis through inhibition of inflammatory mediators and improvement of microcirculation disorders respectively.
基金Scientific Research Fund of national Ministry of Health,No.98-1-173
文摘AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing,the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.RESULTS: ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E.coli M15 and expressed. The expressed ODC protein was verified with Western blotting.CONCLUSION: The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.
基金Supported by the Scientific Research Fund of National Ministry of Health, No. 98-1-173
文摘AIM: To investigate the ornithine decarboxylase (ODC)gene expression in colorectal carcinoma, ODC mRNA was assayed by RT-PCR and ODC protein was detected by a monoclonal antibody against fusion of human colon ODC prepared by hybridoma technology.METHODS: Total RNA was extracted from human colorectal cancer tissues and their normal counterpart tissues. ODC mRNA levels were examined by RT-PCR.ODC genes amplified from RT-PCR were cloned into a prokaryotic vector pQE-30. The expressed proteins were purified by chromatography. Anti-ODC mAb was prepared with classical hybridoma techniques and used to determine the ODC expression in colon cancer tissues by immunohistochemical and Western blotting assay.RESULTS: A cell line, which could steadily secrete antiODC mAb, was selected through subcloning four times.Western blotting reconfirmed the mAb and ELISA showed that its subtype was IgG2a. RT-PCR showed that the ODC mRNA level increased greatly in colon cancer tissues (P＜0.01). Immunohistochemical staining showed that colorectal carcinoma cells expressed a significantly higher level of ODC than normal colorectal mucosa (98.6±1.03%vs 5.26±5%, P＜0.01).CONCLUSION: ODC gene overexpression is significantly related to human colorectal carcinoma. ODC gene expression may be a marker for the gene diagnosis and therapy of colorectal carcinoma.
