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 investigate the effects of catalase on expression of cytokines and activation of nuclear factor-?B in the intestinal mucosa in rat with ulcerative colitis(UC). Methods UC was reproduced in rats with the oral innoculation of dextran sulfate sodium (DSS).The expressions of TNF-?, IL-1? and IL-8 in the intestinal mucosa of rats were respectively determined by a semi-quantatitive assay, RT-PCR. The activation of NF-?B in the intestinal mucosa were assessed with electrophoretic mobility shift assay (EMSA). Results Comparing the mice challenged with DSS alone and to those treated with catalase, both symptoms and the lesions in the colonic mucosa were milder in the animals pretreated with catalase during the induction of colitis than that of control group and catalase treatment after induction of colitis group. Furthermore, the expressions of TNF-?, IL-1?, IL-8 were significantly down-regulated(P

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.

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

To study adhesion of human bifidobacterial strains to cultured human intestinal epithelial cells in vitro mediated by purified adhesin. Results showed that the binding of human bifidobacterial strains to human intestinal epithelial cells appeared to be specific. The adhesion was time and dose-dependent.These data suggested adhesion of human bifidobacterial strains to cultured human intestinal epithelial cells in vitro can be mediated specifically by purified adhesin.

Objective To evaluate the expression of Fas Ligand (FasL) on gastric T cell during H.pylori infection and its cytotoxicity to gastric epithelial cells. Methods FasL protein expressions on mono nuclear cells of gastric mucosa with and without H.pylori infection were examined by immunohistochemistry assay. FasL mRNA expressions were detected in gastric T cell lines isolated from H.pylori infected and uninfected gastric biopsies by RT PCR. The function of FasL expressed by gastric T cells to induce apoptosis in Fas bearing cells was determined by a co culture bioassay (JAM), while the Fas mediated apoptosis of gastric epithelial cells induced by gastric T cells were also evaluated by the same assay. Results FasL was expressed by the mononuclear cells accumulated in gastric mucosa with H.pylori infection, whereas the few mononuclear cells presented in uninfected gastric tissues were negative for FasL. FasL mRNA was detected in gastric T cells isolated from H.pylori infected gastric biopises, while that of uninfected gastric T cell lines was either negative or weak positive. Gastric T cells were capable of inducing apoptosis of Fas positive jurkat cells, while this could be blocked by Fas blocking antibody, ZB4. Conclusions T cells presented in gastric mucosa during H.pylori infection could damage gastric epithelium through Fas/FasL interaction.