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.

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.

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.

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

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 antitumor effects of transmembrane TNF-? and secretory TNF-? in vivo. Methods: Three types of TNF-? cDNA plasmids (wild type TNF-?; transmembrane TNF-? mutant; secretory TNF-? mutant) were directly injected into tumor-tearing mice. Results: The three types of TNF-? could be expressed by tumor cells and all of them could inhibit evidently the rate of tumor growth. The tumor regression after treatment with transmembrane TNF-? mutant at the early stage was more significant than that with the other two types of TNF-?( P

Objective: To observe the inhibiting effects of an antisense u-PAR vector on invasion by highly invasive PC-3M cell subclones. Methods: The effects of an antisense vector on invasion by highly invasive PC-3M cell subclones were observed and compared in vitro by monolayer invasion assay and soft agar clone. Then, both a quantitative RT-PCR and zymography were used to exam the effects of the antisense u-PAR on activity of MMP-9 in those highly invasive cell subclones. Furthermore, the tumorigenesis rate and invasions by the cell subclones with or without the antisense u-PAR were observed in nude mice. Results: It is found that the speed of growth in vitro was slowing down by highly invasive PC-3M cell subclones transfected with the antisense u-PAR, and the ability of anchorage-independent growth of those cell subclones was also decreasing sharply,and the inhibiting rate was 79% and 60%, respectively. Although the antisense u-PAR didn′t change MMP-9 gene transcription, but they could inhibit the activation of MMP-9 of highly invasive PC-3M cell subclones. Moreover, the tumorigenesis rate of the cell subclones with the antisense u-PAR decreased and the growth of a neoplasm also slowed down. The t tests showed the difference between experimental and control groups reached statistical significance ( P

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.