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Objective To study the death mechanism of LoVo cells transfected with retroviral vector carrying cytosine deaminase (CD) suicide gene(G1CEACDNa). Methods Plasmid G1CEACDNa was transferred into the LoVo cells using liposomes method. RT-PCR was performed to detect CD mRNA expression using the total RNA extracted with TRIzol reagent. After exposed to 5-FC (1mmol?L -1 ), the cell growth inhibition rate and the “bystander effect” were determined by MTT, the cell ultramicroscopic structure was observed by electron microscope, and cell apoptosis was detected by flow cytometry. Results RT-PCR detection showed that there was 1.5kb band of CD product. The growth of the trasfected LoVo cells started to be inhibited after exposed 5-FC for 48h, and the inhibition rates at 72h,96h and 120h were 30%,50% and 80%, respetively.Electron microscope examination showed that there were apoptotic body. Meanwhile,a few cells showed necrosis alteration.Flow cytometry analysis showed that a few cells appeared apoptosis after exposed to 5-FC for 48h, the apoptotic rate and the necrosis rate at 72h and 96h were 23.8%, 35.1% and 20.4%, 26.0%, respectively. Conclusion The death mechanism of LoVo cells transfected with plasmid G1CEACDNa followed 5-FC treatment was involved in both necrosis and apoptosis. Apoptosis possibly was a mechanism of the bystander effect.
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Objective To study the mechanism of SW480 cell line death caused by transfection of retroviral vector-G1CEACDNa.Methods Plasmid G1CEACDNa was transferred into the SW480 cell line using liposomes method. RT-PCR was performed to examin the expression of CD gene indrectly.The cell growth curve was measured by means of cell counting. When the CD+SW480 cells were exposed to 5-FC (1mmol/L), the growth inhibition rate and the "bystander effect" were detected by MTT method.The ultrastructure was observed by electron microscope.Apoptosis was verified by flow cytometer .Results The product of RT-PCR showed a band at 1.5kb on the photo of electrophoresis. The growth of CD+ SW480 cells was inhibited 24h after administrating 5-FC,and the inhibition rates at 72h,96h,120h were 30.0%,50.0% and 80.0%,respectively.Apoptosis cells in different phases and apoptotic bodies in the field of electron microscope were observed. Meanwhile ,a few cells showed necrosis.Flow cytometer verified that a few cells appearred apoptosis 48h after exposed to 5-FC (1mmol/L), the apoptosis rate and the necrosis rate at 72h,96h were 20.2%,30.7% and 19.6%,21.1% respectively.Conclusions The death mechanism of SW480 cells transfected with G1CEACDNa followed by 5-FC treatment includes both necrosis and apoptosis.Apoptosis is possibly the bystander effect.
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Objective To investigate the effect of thermochemotherapy with 5 fluorocytosine(5 FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue specific cytosine deaminase (CD) in vitro.Methods Recombinant retroviral vector G1CEACDNa, in which CD gene was controlled under the CEA promoter, was introduced through liposome technique to the human colorectal carcinoma cell line SW480, and then the cells were selectively cultured in G418. The proliferated colonies were treated with the combined therapy of 5 FC and hyperthermia at a 43℃ over 30 min for 3 times. RT PCR was performed to analyze the expression of CD gene in target cells after thermochemotherapy. Results The CD gene expressed steadily in the target cells after 3 times of hyperthermic treatment. After the transfection of CD gene, SW480 CEACD cells were more sensitive to 5 FC than their parental cells (P
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Objective To investigate the diagnosis and treatment of cystic adenoid tumor of the pancreas. Methods Retrospective analysis was made on the clinical data of 11 cases of pancreatic cystic neoplasms confirmed by operation and pathology at our hospital from 1978 to 1998. Results Cystic adenoid tumor of the pancreas is more likely to occur in young females with middle age, clinical presentations were as follows: a long-term persistent mass and distending pain in the epigastrium. Utrasounography, CT and ERCP could contribute to the diagnosis. Of the 11 cases, pancreaticoduodenectomy was performed in 2, distal pancreatectomy and splenectomy in 6, local excision of the mass in 2, and loop type cystojejunostomy in 1. The rete of resection was 90.9%. Of them, 1 case died of abdominal abscess and septicemia postoperatively. All the 10 cases who survived postoperatively were followed up. Among the 10 cases, 1 case of pancreatic cystoadenoma died of cardiac infarction 3 months after operation, and 1 case of cystoadenocarcinoma died of metastasis 51 months after tumor excision. The other 8 cases still alived well now. Conclusions Cystic tumor of the pancreas should be suspected when young or middle aged female patients have a long term big mass in epigastrium or left upper quadrant. Ultrasonography and CT scan are helpful in the diagnosis of the tumor including cystoademoma and cystoadenocarcinoma, Although the volume of the tumors may be very big, they often have a intact capsure,so easy to excise. The excision of the total tumor and some arounding pancreatic tissue is the first choice of the treatment for cystic tumor of the pancreas, and the curative effectiveness is satisfctory.
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0.05). RFA showed a marked short term therapeutic effect for patients with PHC.Conclusions After RFA treatment of patients with PHC,the serum levels of TNF and AFP decreased,and this suggests an augmentation of organism immune function. The serum levels of TNF and AFP can assist in the assessment of the therapeutic effect and prognosis of primary hepatic carcinoma.
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Objective To introduce how to improve the operation quality and clinical effect of pancreatoduodenectomy (PD). Methods The morbidity and mortality of 126 patients with pancreatic head cancer or peri ampullary cancer treated by PD of traditional Child method and modified Child method ( pancreatic stump closing style pancrea to jejunum anastomosis) in our department from 1973 to 2002 were analyzed retrospectively. Results In modified Child method group, the morbidity of pancreatic leakage and the mortality of the operation were 2.4% and 4.7% respectively, which were significantly lower than those in the traditional method group(P