MG132 Induced Apoptosis Pathway in HL-60 Cells and Impact of Allogeneic Mixed Lymphocyte Reaction

Objective： To investigate the proteasome inhibitor MG132-induced apoptosis pathway in HL-60 cells and the role of allogeneic mixed lymphocyte reaction. Methods： Cell apoptosis was analyzed by flow cytometry. The expressions of p21 protein, p27 protein and p53 protein in HL-60 ceils treated with MG132 were measured by Western blot. The proliferation of, peripheral blood mononuclear cells （PBMNCs） after treatment with 75 Gy irradiated HL-60 cells treated with MG132 was measured with CCK-8. Results： High-dose MG132 induced apoptosis in HL-60 cells. No significant change was observed in MG132-induced apoptosis after inhibiting caspase-8 and caspase-9 pathway. The expressions of p21 protein and p27 protein increased in MG132-induced apoptosis. HL-60 cells treated with low-dose MG132 improved the proliferation of PBMNCs from healthy volunteers. Conclusion： High-dose MG132 induced apoptosis and directly killed HL-60 ceils. MG132 induced apoptosis in a caspase-8- and caspase-9-independent pathway, p21 protein and p27 protein were involved in MG132-induced apoptosis in HL-60 cells. HL-60 cells treated with Low-dose MG132 improved the effect of promoting the proliferation of PBMNCs from healthy volunteers.

Numerical simulation of micromixing effect on the reactive flow in a co-rotating twin screw extruder

To control the multicomponent reactions in extrusion, reactive-mixing flow in a co-rotating twin screw extruder was numerically studied in the present paper. Effects of initial species distribution, rotating speed and flow rate on a competitive-parallel reaction were investigated and the relationship between mixing and reactions was discussed from the view of chemical reaction engineering. The simulation results show the studied operational parameters, which determine residence time distribution, earliness of mixing and segregation degree of reactive-mixing flows, affect the local species concentration and reaction time and hence have significant influences on the reaction extent. Orthogonal test was adopted to clarify the significance of operational parameters.The analysis shows that initial species distribution and flow rate are the most important factors in the control of reaction extent, and effect of rotating speed is conditional depending on the micro-mixing status of the fluid.

IMPAIRED AUTOLOGOUS MIXED LYMPHOCYTE REACTION CORRELATED WITH DECREASED EXPRESSION OF HLA-Ⅱ ANTIGENS ON MONOCYTES IN PATIENTS WITH MYELOID LEUKEMIA

The proliferative response of T-cells to autolo-gous non-T-cells is referred to as the autologous mixed lymphocyte reaction (AMLR). Recent studies have suggested that AMLR represents a mechanism of immune regulation in vivo. We investigated AMLR in patients with acute- and chronic myeloid leukemia (AML and CML). AMLR was found to be significantly depressed (P<0.001) in AML patients (n=17, cpm=532±95) and CML patients (n=13, cpm=688±99) when compared with that of their healthy HLA-identical siblings serving as controls (n=17, cpm=4152±619 and n=13 cpm=4086±421, respectively). In order to understand the cellular basis of the defective AMLR in patients with AML end CML, we performed mitogen-treated T-cell cultures analysis of T-cell subsets and HLA-Ⅱ antigen detection on monocytes. The results indicated that the defect of AMLR in patients resided at the stimulator monocyte level rather than at the responder T-cell level. Enumeration of monocytes reactive with monoclonal antibody Tu22, which recognizes determinants of HLA-DQ, demonstrated that ML patients had a significantly decreased (P<0.091) number of circulating Tu22+ monocytes when compared with normal controls. These studies suggest that a deficiency of HLA-DQ+ monocytes contributes to the depression of AMLR in ML and possibly underlies the abnormalities of immune response present in this disease.

STUDIES ON THE CHROMOPHORIC REACTIONS OF MIXED POLYNUCLEAR COMPLEXES OF RARE EARTH-ARSENAZO Ⅲ-COPPERO-PHENANTHROLINE

In an acidic solution the mixed polynuclear complexes could be formed by association of rareearth-arsenazo Ⅲ complexing anions with copper o-phenanthrolinate cations.In this paper the formationconditions of lanthanum-arsenazo Ⅲ-copper-o-phenanthroline complex and its related reaction mechanismhave been studied in detail.The complex exhibits an absorption maximum at 663 nm and its apparent molarabsorptivity is found to be 1.22×105L·mol-1·cm-1.The molar ratio of the components in the complex isestimated to be La:arsenazo Ⅲ:Cu:Phen=1:1:1:3.Its possible molecular formula can expressed as[Cu（Phen）3]（La-arsenazo Ⅲ）.The reaction is used to determine micro amounts of lanthanum（or rareearths）in copper containning synthetic samples and satisfactory results are obtained.

Temperature-dependent Hydrothermal Synthesis of Two Distinct Three-dimensional Copper Complexes

Two distinct copper coordination polymers, namely [Cu^Ⅱ2（2,5-pydc）2（bpp）2]·H2O（1） and Cu2^ⅠCu^Ⅱ（2,5-pydc）2（bpp）2（2）（2,5-pydc = pyridine-2,5-dicarboxylic acid, bpp = 1,3-bi（4-pyridyl）propane）, have been successfully synthesized through hydrothermal conditions under different temperatures. Single-crystal X-ray structural analysis revealed that both complexes 1 and 2 are 3D frameworks. Complex 1 is an 8-connected 2-fold interpenetrating network based on [Cu（2,5-pydc）]4 molecular building block（MBB）, and also can be simplified as a 4-connected net if the Cu（Ⅱ） ion is regarded as an independent node, whereas 2 shows a（4,4）-connected non-interpenetrated framework which contains mixed valence Cu（Ⅰ/Ⅱ） centers. The results demonstrate that temperature plays a significant role in the final structures of the complexes.

Clinical Grade of Gerneration of Dendritic Cells for Immunotherapy

In order to develop a protocol for clinical grade generation of dendritic cells (DCs) for cancer immumotherapy, aphereses were performed with the continuous flow cell separator and mate- rials were derived from 10 leukemia patients that had achieved complete remission. Peripheral blood monocytes were cultured in vitro with GM-CSF, IL-4 for 6 days, then TNF-α (the TNF-α group) or TNF-α, IL-1β, IL-6, PGE2 (the cytokine mixture group) were added to promote maturation. Cell number was counted by hematology analyzer, and phenotype study (CD1a, CD14, CD83) was carried out by flow cytometry, and the function of DCs was examined by mixed lymphocyte reaction. The results showed that (0.70±0.13)×107/mL (the TNF-α group) and (0.79±0.04)×107/mL (the cytokine mixture group) DCs were generated respectively in peripheral blood obtained by leucapheresis. The phenotypes were as follows: CD1a+ (74.65±4.45)%, CD83+ (39.50±4.16)%, CD14+ (2.90±1.76)% in TNF-α group, and CD1a+ (81.86±5.87)%, CD83+ (81.65±6.36)%, CD14+ (2.46±1.68)% in the cyto- kine mixture group. It was concluded that leucapheresis may be a feasible way to provide large num- ber of peripheral blood monocytes for DC generation, and combined administration of TNF-α, IL-1β, IL-6, and PGE2 may greatly promote maturity.

Mixed lymphocyte reaction induced by multiple alloantigens and the role for IL-10 in proliferation inhibition

The frequency of T cells that can respond to alloantigens is unusually high.It remains unclear how T cells would respond when stimulated by multiple major histocompatibility complex(MHC)disparate alloantigens in the same cultures.In this report,we examined potential interactions of T cell clones that were stimulated simultaneously by two sets of complete MHC disparate alloantigens using mixed lymphocyte reaction(MLR).In this assay,we observed that proliferation of B6 lymphocytes(H-2b)stimulated by both BALB/c(H-2d)and C_(3)H(H-2k)allogeneic cells was not increased but rather reduced as compared to B6 cells stimulated with either BALB/c or C_(3)H allogeneic cells.Interestingly,interleukin(IL)-10 expressions at both protein level and mRNA level was signifi cantly increased in cultures stimulated with the two MHC alloantigens,while IL-2,tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β1 production did not show any differences.In addition,Foxp_(3) mRNA expression was comparable amongst all groups.In conclusion,we observed an inhibitory effect in T cell proliferation in response to multiple MHC mismatched alloantigens in MLR,and this effect might be associated with the upregulation of IL-10 expression.

Mixed Reactions

UN Secretary General Ban Kimoon, while welcoming the diplomatic efforts by Turkey and Brazil, reiterated hiscall to Iran to show greater transparency on the nuclear fuel swap deal. He said

MIXED REACTION TO WORKERS' RIGHTS LAW

Legal protection for workers’ rights, adopted this year, has been applauded by workers but criticized by management Few Chinese laws have taken as longto draft as the Labor Contract Law,which after 11 years in the draft

Carboxymethylpachymaran Enhances Immunologic Function of Dendritic Cells Cultured in Two Kinds of Hepatoma Carcinoma Cell Line's Supernatant via Nuclear Factor κ B/Rel Pathway

Objective：To study the immunologic function of dendritic cells（DCs） cultured in two kinds of hepatoma cell line＇s supernatant and the enhancing effects of carboxymethylpachymaran（CMP） on DCs.Methods：DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor （GM-CSF） and interleukin（IL）-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines（HepG2 and HepG2.2.15） were collected for condition medium（CM） according to a volume ratio of supernatant to incomplete RPMI-1640 medium,which was 3：1.CMP was dissolved in incomplete RPMI-1640 medium.Experimental groups were divided according to the culture medium,either CM or with CMP in it.DCs subsets CD83,CD86,CD1a,and d-related human leukocyte antigens（HLA-DR） were analyzed by flow cytometry.The proliferation ability of allogeneic T cells in mixed lymphocyte reaction（MLR） stimulated by DCs was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide（MTT） analysis.IL-12p70,interferon-γ（IFN-γ）,and nuclear factor k B（NF- k B） were detected by enzyme-linked immunosorbent assay analysis.Results：The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group（P0.01）.Compared with the normal group,groups treated with CMP showed a higher expression level of DCs subsets,lymphocyte reproductive activity,as well as IL-12 and IFN-γsecretion levels.Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γsecretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group（P0.05）.Compared with the normal group,the expression level of NF-k B in DCs nuclear was higher in CMP groups but lower in two CM groups（P0.05）.After CMP was added,the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added（P0.05）.However,there was no significant difference between the two CM groups（P0.05）.Conclusions：Two kinds of hepatoma cell line＇s supernatant can inhibit the immunologic function of DCs.This suppressive effect may be related to the inhibition of NF-κB/Rel pathway.CMP may up-regulate the DCs function by activating the NF-κB/Rel pathway.