Daily i.v. injection of 4ml of the traditional Chinese medicine antiinflammatio No.6(AI6) to rats for 3 days resulted in a marked acceleration of chemotaxis (Cht) of neutrophil granulocytes(NG). Daily i.m. injection of streptomycin(SM) and chloromycetin(CM), each in a dose of 50mg per animal, given to rats for 3 days led to a marked depression of Cht of NG, while combined with daily i.v. injection of 4ml of AI6 caused not only a complete cancelling of the suppressive effect of the antibiotics on NG Cht, but also a marked acceleration of NG Cht. Combined application of AI6 and certain antibiotics in the clinical treatment of certain acute bacterial infections is suggested.

Recent studies have shown that the the nervous system possesses immunoregulatory function. We investigated the effect of substance P(SP) a neuropeptide on the expression of HLA-DR and HLA-DQ antigens on normal human monocytcs of the peripheral blood. APAAP (alkaline phosphatase-antialkalinc phosphatasc) enzyme immunoassay was adopted to detect the HLA class Ⅱ molccults, using monoclonal antibodies Tu36, Tu22, in the reaction system. The results, showed that SP at concentrations higher than the physiological serum level (10~(-10)mol/L, 30~(-8)mol/L, 10~(-6)mol/L) promoted the expression of HLA-DQ antigen on monocytcs significantly in vitro (P0.05). The maximal effect was observed at 10~(-8)mol/L. Expression of HLA-DR antigen, however, was not influenced by SP in the same range of SP concentrations. It is reasoned by the authors that the immunosupprcssivc effect of excessive glucocorricoids during stress might partly be counterbalanced by the uprcgulatory action of increased blood SP on the immune function.

Objective　To construct a recombinant plasmid pcDNA3-sHLA-G1 expressing soluble HLA-G1. Methods　 Total cell RNA was extracted from the cell line Jeg-3 and the cDNA was amplified by RT-PCR； The cDNA fragment was inserted into the eukaryotic expressing vector pcDNA3 and the recombinant plasmid was identified by restriction endonucleases digestion and sequencing. Results　 After restriction endonucleases treatment and sequencing, it was confirmed that the pcDNA3-sHLA-G1 had been constructed successfully. Conclusion　 In this study, the recombinant plasmid pcDNA3-sHLA-G1 had been constructed successfully.

Transmembrane TNF-? is an integral protein expressed on the surface of avtivated monocytes/macrophages. In order to define the interrelation between TM-TNF and biomembrane, we used an in vitro translation system and microsomal membranes to engender expression models of two types of TM-TNF; TM-TNF anchored to microsomes and naked 26kD TNF without membranes. When the cytotoxic activity of these two types of TNF was compared, the synthesized 26kD TNF was found to exert its effects only in the presence of microsomal membranes. With the use of indirect immunofluorescence technique, we further studied the binding of the in vitro translated 26kD TNF with TNFR on the membrane of HL60 cells. It was found that only TM-TNF attached to the microsomes could bind effectively with TNFR, while naked 26kD TNF could not. These results suggest that binding with biomembrane may be the prerequisite for 26kD TNF to fold properly for displaying its biological effects. In case 26kD TNF is freed from membrane, it can no more bind with TNFR, there by leading to loss of its cytotoxic effect.

In order to establish stable high expression cell lines, the eukaryotic expression vector pIRES2EGFP and recombinant plasmid pIRES2EGFP-TIM-3 were transfected into mammalian cells CHO by Lipofectamine. The transfected cells were cultivated under selective growth medium including G418 and green fluorescent protein (GFP) positive cells were sorted by FACS. Simultaneously, growing transfectants were selected only by G418 in the medium. The GFP expression in stably transfected cells was detected by FACS. Under selective growth conditions with G418, the percentage of GFP positive cells was reduced rapidly and GFP induction was low. In contrast, the percentages of GFP positive cells were increased gradually after FACS. By 3 rounds of GFP selection, the stable high expression cell lines were established. Furthermore, using FACS analysis GFP and the target protein TIM-3 co-expression in the stable transfectants cultured in nonselective medium was detected. Theses results demonstrated that the stably transfected cell lines that express high titer of recombinant protein can be simply and fleetly obtained by using GFP and selective growth medium.

Objective:To study the effects of secreted stem cell factor(S SCF) and transmembrane SCF(TM SCF) on the biological functions of mast cells,including proliferation,release of histamine and secretion of cytokine to explore the biological characteristics of the two forms of SCF.Methods:MTT,fluorescence technique,in situ hybridization were used to detect the several biological functions of mast cells,such as proliferation,release of histamine and the transcription of TNF ? mRNA.Results:It was shown that S SCF could significantly promote the proliferation of mast cells,release of histamine and TNF ? mRNA transcription,while TM SCF had only a weaker positive effect on histamine relase and no any effect on proliferation and TNF ? mRNA transcription.Conclusion:The biological actions of TM SCF on mast cells are different from that of S SCF.

Objective:To determine the region of human transmembrane tumor necrosis factor alpha (TM TNF?),essential for cytotoxic activity against mouse L929 cell.Methods:Single amino acid substituted TM TNF? mutant proteins(muteins) were produced by in vitro transcription linked translation techniques.An expression cDNA for TM TNF? was site directed mutagenized by recombinant PCR.7 single amino acid substituted TM TNF? muteins were generated and assayed for cytotoxic activity.Results:The cytotoxic activities of TM TNF? muteins,eg,TM TNF? -71/Lys was

The activity of the NK cells in patients with preeclampsia was studied to investigate the pathogenesis of preeclampsia. By using MTT and 51Cr releasing technique, the proliferation and killing ability of the NK cells in maternal and umbilical blood from preeclampsia patients (n = 18) and normal third trimester pregnant women (n = 18) were detected. The NK-92 cell line was as the positive control. The results showed that the NK cell counts of umbilical blood in preeclampsia patients and normal third trimester pregnant women were significantly greater than those of maternal blood (both P<0.05). Compared with that in normal third trimester pregnant women, the proliferative ability of the NK cells in preeclampsia patients was apparently increased (P<0.05). Compared with that in maternal blood, the proliferative ability of the NK cells in umbilical blood from both preeclampsia patients and normal third trimester pregnant women was dramatically increased. The killing ability of the NK cells in preeclampsia patients was significantly higher than that in normal third trimester pregnant women (P <0.05). It was suggested that both number and function of the NK cells in preeclampsia women were increased, and that in umbilical blood was greater than that in maternal blood, speculating that the function of the NK cells may affect the maintenance of the maternal and fetal immune tolerance during pregnancy.
Cytotoxicity, Immunologic/*immunology ; Fetal Blood/cytology ; Immune Tolerance ; Killer Cells, Natural/*immunology ; Killer Cells, Natural/pathology ; Pre-Eclampsia/blood ; Pre-Eclampsia/*immunology ; Pregnancy Trimester, Third

Objective Objective In order to better keep up with the development of transplantation immunobiology.we established and compared two types of mouse heterotopic heart transplantation,and hope to help further organ transplantation studies.Methods According to the surgical procedures of Ono's type and Chen's type of mouse model of heterotopic heart transplantation with some modification,we performed celiac and cervical heteropotic heart transplantation between iso-strains and hetero-strains,and compared the operation suecess rate,operation time,allografi survival time,and histopathology of those establishment methods.Results The success rates of mouse celiac and cervical heterotopic heart transplantation were 86.7% and 83.3%,respectively,with a non-significant difference(P>0.05) between the two methods of operation regarding the total operation time,survival time of the allografts,and histopathological findings.Conclusions Based on the mastery of microsurgical techniques,the two models of heterotopic mouse heart transplantation can be established equally,and either of them can be considered depending on the particular requirements of studies.

AIM: To compare the tumorigenecity of H22 cells transfected with TNF-? gene and its mutants (secreted TNF-? mutant, S-TNFm, transmembrane TNF-? mutant, TM-TNFm and wild type of TNF-?, Wt-TNF) in vivo . METHODS: Three kinds of mouse liver cancer cell line H22 expressing TNF-? and its two mutants were mixed with untransfected H22 at different effector/target ratio separately. The growth of tumor was examined after injection of 2.5?10 5 (100 ?L) mixed H22 tumor cells into mice. The lymphocyte infiltration in the site of tumor and the expression of Fas on tumor cells were detected by immunohistochemistry. RESULTS: The tumorigenecity of H22 cells transfected with TNF-? gene and its mutants was significantly weakened ( P