Utilizing higher expressing vector and a synthetic oligonucleotide linker,a new expressing clone, pBMhIL 4, has been constructed in our laboratory. The intact human rIL-4 protein molecule was expressed under the control of pL promoter in E. coli. The gene encoding protein was expressed in the plasma of E. coli in the form of inclusion bodies, the expressed IL-4 constituting about 5-10% of the total cellular proteins by SDS-PAGE analysis. The molecular weight of human rIL-4 was about 15KD. After extraction and renaturation, the yieldedsoluble human rIL-4 exhibited biological activity. 1?10~6 units of human IL-4 were produced from 1 liter of bacteria extracts assassed by the assay of its TCGF activity. Using 3.5M Guanidine hydrochloride and 0.25% Triton X100 to lyze inclusion bodies resulted in higher yield and higher biological activity.

Utilizing Polmerase Chain Reaction (PCR) and recombination DNA techniques with synthetic oligonucleotide primers,a high level expressing clone for human IL- 6, pBMhIL6, was constructed successfully in our laboratory. The intact human rIL-6 protein molecular was expressed under the control of p_Rp_L promoters, synthetic SD sequence and the terminal codon TAA which replaced TAG of original human IL-6 in E. coli. the expressed human rIL-6 protein, molecular wight 21 KDa, with specific binding to Anti-IL-6 McAb, constituted about 28% of the total cellular proteins assessed by SDS-PAGE, densitometry analysis and Western Blot assay. The results of study on kinetics of inducing human rIL-6 expression in the defferent E. coli strains and influence of bacteria growth states ingdicate that the higher productive ratio for human rIL-6 inducing expression was obtainde in E. coli DH5a at O. D=0. 7. 5?10~6 unites of human rIL-6 HPGF bioactivities were produced from 1 litre of bacteria extracts assayed by 3HTdR uptake of 7TD1 cell line.

Two hour after LAK—like cells were co—cultured with gastric cancer cells,the mixed sus-pension was spread on slids and dyed with histochemical method.Through analysing the dyedcells,we directly confirmed what subset the LAK—like cells belonged to.The featrue of LAK—like cells adhering to cancer cell was observed under electron micrscope in our experiment.A-mong the lymphoctes which were adherent to cancer cells,the number of CD8~+ cells was in-creased.Most of adherent to cancer cells,the number of CD8~+ cells was increased.Most of ad-herent cells were TAC~+,suggesting that the expression of TAC antigen plays an important rolein cell mediated cytotoxity.

Objective To construct a highly expresses catalase of Helicobacter pylori (H.pylori) and to test it's activity. Methods The catalase DNA was amplified from H.pylori chromosomal DNA by PCR techniques, inserted into the prokaryotie expression vector pET 22b(+) and then transformed into the BL21(DE3) E.coli strain which expressed catalase recombinant protein. Then the activity of H.pylori catalase was assayed by the Beers and Sizers. Results DNA sequence analysis showed that the sequence of catalase DNA was the same as that published by GenBank. The catalase recombinant protein amounted to 24.4% of the total bacterial protein after inducing with IPTG for 3 hours at 37?C and the activity of H.pylori catalase was high in the BL21 (DE3) E.coli strain. Conclusions A clone of expressing high activity H.pylori catalase has been obtained, and thus a good foundation for further study has been laid.

Objective To study the changes of serum soluble tumor necrosis factor receptor Ⅰ (sTNFR Ⅰ) and immunosuppressive acidic protein (IAP) of patients with pancreatic cancer before and after intra arterial chemotherapy and evaluate their significance. Methods The levels of sTNFR Ⅰ and IAP of 55 cases with pancreatic cancer before and after intra arterial chemotherapy were measured by enzyme linked immunosorbent assay (ELISA) and one direction immunodiffusion test respectively and compared them with those of healthy controls. Results The levels of sTNFR Ⅰ and IAP of patients with pancreatic cancer before intra arterial chemotherapy were higher than those of healthy controls ( P

Objective To construct a recombinant strain which expresses adhesin AlpA of Helicobacter pylori(H.pylori) and to study the immunogenicity of adhesin AlpA. Methods The adhesin AlpA DNA was amplified by PCR and inserted into the prokaryotie expression vector pET-22b(+)and expressed in the BL21(DE3) E.coli strain. Adhesin AlpA immunogenicity was studied by Western blot test. Results DNA sequence analysis showed that the sequence of adhesin AlpA DNA was almost the same as that of the published by GenBank. The adhesin AlpA recombinant protein accounted for 31.9% of the total bacterial protein. Western blot analysis of rAlpA confirmed that it could be specially recognized by serum from rabbit immunized with AlpA itself and H.pylori infected patients. Conclusion Cloning and high-expression of adhesin AlpA and primary confirmation of its immunogenicity lay the foundation for H.pylori gene engineering vaccine preparation and adhering mechanism research.

The studies were undertaken on the trauma model of rats,whose right hind legs were amputated,in order to investigate the change in the immune adherence function of red cells,and the effect of treatment with Traditional Chinese Medicine,consisting of a mixture of Shen-er,to decrease immune adherence function of red cells after trauma.The results indicats that after trauma (1) the activity of C3b receptor of red cells decreased;(2) the amount of immune complexes on the surface of red cells increased;(3) the activity of the enhancing factors for the immune adherence of red cells dropped;(4) the activity of the inhibitors for the immune adherence of red cells elevated;(5) there is remarkable interrelation in the changes between the activity of the C3b receptors of red cells and the activity of the inhibitors for the immune adherence of red cells;(6) the mixture of Shen-er could effectively raise the inhibited activity of the C3b receptor of red cells;(7) the mixture could also effectively regulate the activity of the inhibitors of the immune adherence of red cells.

To observe changes in blood pressure of dogs with hemorrhagic shock during infusion of 7 5% NaCl, five mongrel dogs were bled to MAP 5 33～6 7kPa and then kept at this pressure for 1 hour. Each dog received 7 5% NaCl solution (4ml/kg) intravenously and the infusion time was 2 min. The results showed that during the 2 min infusion period the systolic pressure (SP) was obviously higher than that before the infusion ( P

To study the changes in serum immunosuppressive acidic protein (IAP) and T lymphocytes rDNA transcription activity in peripheral blood of patients with pancreatic cancer before and after operation. The level of IAP and T lymphocytes rDNA transcription activity in peripheral blood of 58 patients with pancreatic cancer were measured before and after opreration by one directional immunodiffusion test and a cell image analysis of Ag NORs, respectively, and the results were compared with that of healthy controls. The level of IAP was higher while T lymphocytes rDNA transcription activity was lower before operation compared with healthy controls( P 0 05). The levels of IAP and T lymphocytes rDNA transcription activity were closely related to tumor invasion, metastasis and clinical stages (TNM). Serial determination of the levels of IAP and T lymphocytes rDNA transcription activity might serve as an important index for evalution of tumor invasion and metatasis, and also prognosis for the patients with pancreatic cancer

Aim To investigate roles of ROS in oxaliplatin-induced PUMA expression and apoptosis in colon cancer cells.Methods ROS was used as an oxidative stress in vitro and PUMA expression was determined by Western blot analysis,the apoptosis induced by ROS was assessed by Hoechst 33258 dye staining,the proliferation of the colon cancer cells treated with oxaliplatin and antioxidant was determined by MTT assay.Results ROS induced apoptosis and PUMA expression in colon cancer cells;suppression of PUMA expression by stable transfecting PUMA anti-sense vector decreased apoptosis induced by ROS;oxaliplatin-induced PUMA expression was abrogated by antioxidant and the proliferation of colon cancer cells treated with oxalipla-tin was increased by antioxidant.Conclusions Our data suggests that PUMA and ROS play important roles in oxaliplatin-induced apoptosis.Oxaliplatin induces PUMA expression and apoptosis partly through ROS in colon cancer cells.