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Mouse hybridoma monoclonal antibody is the most commonly used antibody in immunology because of its stable source, easy preparation in later stage and high yield. The traditional time-consuming and laborious hybridoma preparation technology could not meet the growing market demand. In this paper, we describe the rapid preparation techniques involved in antigen design and screening, B cell enrichment and screening, transgenic myeloma cells, fusion technology improvement, positive hybridoma cell screening and rapid detection of monoclonal antibody performance, to provide a reference for rapid preparation of mouse hybridoma monoclonal antibody.
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Animals , Mice , Antibodies, Monoclonal , Antigens , B-Lymphocytes , HybridomasABSTRACT
Objective To investigate the co-transfection of p53 and angiostatin gene in the inhibition of SG7901. Methods Transfected the pVITRO2-hp53-hAS into SG7901 with lipofectamine.After transfection, RT-PCR were used to know whether the aimed gene had been transfected and expressed or not. Cell clones trial, MTT growth curve, cell cycle measuring were used to analyze the differences. Results The cells were suppressed by the two genes and inhibition of the combined genes is more powerful than single one. Conclusion The effection of combined genes pVITRO2-hp53-hAS on SG7901 is stronger than either single one. Combined-gene-therapy is a useful anti-carcinoma method.
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Objective To investigate the function of estrogen in the proliferative and involuting stages of infantile hemangioma. Meothods Expression of estrogen receptor(ER),VEGF and bFGF were detected with SP immunohistochemical method in 42 cases of hemangiomas and 17 cases of vascular malformations.Results The label index(LI) of ER,VEGF and bFGF in the hemangioma was significantly higher than those in the vascular malformation and normal skin(P
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Objective To study the relationship between the expression of chemokines monocyte chemoattractant protein 1 (MCP 1) and the biological behavior of colorectal carcinoma by detecting chemokine mRNA and protein in human colorectal cancerous tissues. Methods The expression of the MCP 1 mRNA was detected in colorectal carcinoma collected freshly from surgical specimens by RT PCR and the expression of the MCP 1 protein was assessed in colorectal carcinoma tissues collected from surgical specimens by immunohistochemistry. Results All the 12 colorectal carcinoma expressed the MCP 1 mRNA, which was detected by RT PCR; MCP 1 protein was detected in 90% of the tumors; The expression of the MCP 1 protein in colorectal carcinoma was correlated with its state of metastasis and the Dukes' stage. Conclusions The expression of chemokine MCP 1 in colorectal carcinoma may influence its biological behaviour.
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Purpose:To study the expression and clinical significance of p27 in esophageal carcinoma. Methods:Immunohistochemical staining was used to determine the expression of p27 in 32 cases of normal esophageal issues and 88 cases of esophageal carcinoma.Results:The positive incidences of P27 expression in esophageal carcinoma was 40.9% while in normal esophageal issues it was 75.0%. The expression of p27 was correlated to degree of differentiation of carcinoma and lymph node metastasis, but not to length, depth of invasion and TNM stage. The p27 negative group was seen to have the worst survival rate among all groups.Conclusions:The results suggest that p27 gene plays an important role in carcinogenesis and progress of esophageal carcinoma. Expression of P27 may prove to be valuable in prognosis assessment of esophageal carcinoma.
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Objectives:To investigate the therapeutic effect of somatostatin and parenteral nutrition on early postoperative inflammatory intestinal obstruction. Methods:16 patients(test group) were treated with somatostatin and parenteral nutrition.Somatostatin was administered in 6 mg doses daily through continuous intravenous drip and the formula of parenteral nutrition was provided according to clinical routines.The therapeutic effect was compared with that of previous 13 cases(control group) without use of somatostatin and parenteral nutrition. Results:The rates of transferring to operation in the test group and the control group were 12.5 % and 61.5%( P
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Objectives:To investigate the effect of GH added early postoperative enteral nutrition on protein metabolism and immune function. Methods:Thirty six patients undergoing total or subtotal gatrectomy were randomly divided into three groups:traditional entertal nutrition (TEN),early postoperative enteral nutrition(EPEN),and GH added early postoperative enteral nutrition(GH+EPEN).Nutritional support lasted for 7 days.GH daily dose was 8 IU.Nutritional markers and immune markers were detected in several time points. Results:On postoperative day 4,the levels of Alb,Tf,TCL,and CD4/CD8 were all significantly increased in GH+EPEN as compared with those of TEN(all P
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Objectives:To investigate the effect of growth hormone(GH) and parenteral nutrition (PN) on protein metabolism in patients with cirrhosis of liver. Methods:The test group received GH and PN,and the control group received PN only.All the treatments were maintained for 7 days.The serum levels of Alb,TFN and PA were detected on the day before the treatment and the day 3,5 and 7 after the treatment. Results:As compared with the day before the treatment,the levels of Alb,TFN and PA on the day 3,5 and 7 in the two groups were all increased.However,the changes in the test group were greater than those in the control group.As compared with the control group,the level of Alb on the day 7 and the levels of TFN and PA on the day 5 and 7 in the test group were significantly higher. Conclusions:The therapeutic value of combined use of GH and PN is much better than that of single PN in patients with cirrhosis of liver.
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Objective To study the diagnosis and treatment of gastric stromal tumor(GST). Methods The clinical manifestation,pathological features ,diagnosis and treatment of 14 patients with GST were retrospectively analysed . Results Of the 14 GST,7 located in the fundus and the body of the stomach, 7 in the antrum of the stomach. The main symptom was abdominal pain, All the 14 GST were diagnosed by endoscopey, barium, ultrasound and CT. All patients were treated by excision. Histological diagnosis was as benign tumor in 8(57.1%)cases, uncertain types in 5 (35.7%)cases , and malignant tumor in 1 ( 7.1 %)case. There were local recurrence in 3 cases and death in 1 case after the operation. Conclusions The complete local excision is recommended for GST patients. Long term postoperative follow up is necessary for patients with GST. Reexcision may be helpful to the patients with recurrence or metastasis.