Objective To investigate effect of first, second, and third trimester placental factors (PF) on CD4, CCR5, and CXCR4 expression in human peripheral blood lymphocytes (PBLs), and to explore their influence on human immunodeficiency virus (HIV) vertical transmission.Methods Human peripheral blood mononuclear cells (PBMCs) were treated with first, second,and third trimester PF (concentration 25%) respectively for 24 hours. The expression of CD4, CCR5,and CXCR4 in PBLs, and the percentages of CCR5+, CXCR4+,and CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes were determined with flow cytometry.Results All trimester PFs reduced CCR5 expression in PBLs. The efficiency of the first trimester PF was higher than that of the second and third trimester PF. The percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of PF groups was significantly lower than that of the control group, and the percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the second and third trimester group. The percentages of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of PF groups were significantly decreased as compared with the control group, and the percentage of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the third trimester PF group.Conclusion PF can reduce the expression of CCR5 in human PBLs and peripheral blood CD4+ lymphocytes, indicating that PF might reduce R5 virus infection via preventing HIV entry, and might play an important role in reducing R5 virus intrauterine infection.

AIM: To study the relation between prolon ge d survival of skin allograft by chuan-ke-zhi (CKZ, drug of Chinese herbal) and C D4+CD25+ regulatory T cells (CD4+CD25+ Tr) in mice. METHODS: Skin allograft and isograft model in mice were establi shed and CKZ was administered by intraperitoneal injection. To observe its influ ence on survival of the graft, three color fluorescent staining together with fl ow cytometry was used to analyze the change of CD4+CD25+ Tr. RESULTS: The survival of skin allograft in CKZ group was signifi cantly prolonged compared to control group, (19.5?2.3) days and (10.2?2.2) days, respectively, P0.05). CONCLUSION: CKZ has an effect of prolonging the survival of skin allograft. Enhancement of CD4+CD25+ Tr might be one of the mechanisms under lying its immunosuppressive effect.

AIM: To study the effect of supernatants from cord blood mononuclear cells (CBMC) and peripheral blood mononuclear cells (PBMC) on infection of HIV-1 in vitro, and develop effectively soluble factors for human acquired immunodeficiency syndrome (AIDS) treatment. METHODS: Different supernatants from CBMC and PBMC activated by PHA for 5 hours and 12 hours were added to cell culture systems between HIV-1ⅢB/H9 cells labeled by calcein-AM and MT-2 cells, then to count the fusion under a reverted fluorescent microscope after 2 hours, respectively, and different soluble factors in supernatants were detected by Luminex 100~ TM . RESULTS: These supernatants from CBMC and PBMC activated by PHA for 5 hours and 12 hours inhibited the formation of fusion, and there is no difference between supernatants collected in CBMC and PBMC at same time point. However, the supernatants collected in 5 hours were more effective to inhibit the formation of fusion than those in 12 hours (P

Objective To investigate the effect of Pingfei Oral Liquid (POL) on the distribution of mast cells (MCs) and the expression of interleukin 6 (IL-6) in the lung tissue of radiation pneumonia rats. MethodsForty-five SD rats were randomized into 3 groups : normal control,model group and POL group. The rat model of radiation lung fibro-sis was set up by a single X-ray dose of 20Gy irradiation over the whole chest of the rats. POL (20 g·kg-1·d-1,once a day, five times a week) was given orally one week before irradiation and the treatment lasted 5 weeks. MCs in the lung tissue were stained with toluidine blue firstly and then were counted 2, 4 and 8 weeks after irradiation. IL-6 protein expression of lung tissue was measured by immunohistochemical assay 8 weeks after irradiation, and mRNA ex-pression was determined with RT-PCR 4 weeks after irradiation. ResultsIt's showed the aggregation of large amount of pulmonary mast cells and increase of IL-6 protein expression 8 weeks after irradiation (P < 0.01).IL-6 mRNA expression in the irradiated lung of rats increased 4 weeks after irradiation (P < 0. 01). POL could reduce the aggrega- tion of MCs (P < 0. 01) and the expression of IL-6 protein (P < 0. 01) and mRNA (P < 0. 05) in the lung tissue. ConclusionPOL can prevent radiation pneumonia in rats by reducing the aggregation of mast cells and inhibiting IL-6 expression in the lung tissue.

This study was aimed to establish a HPLC method for the determination of isogarcinol. Daojin Inertsil WP300 C18 (4.6 mm× 150 mm, 5μm) was employed with methanol and water (75?25) as the mobile phase. The flow rate was 1.0 mL·min-1. The column temperature was set at 40°C. The detection wavelengthγ was set at 277 nm. The results showed that the linear range of isogarcinol was 0.005 7-0.039 9μg. The average recovery rate was 99.58%. The relative standard deviation (RSD) was 1.25%. The contents of isogarcinol inGarcinia mangostana,Garcinia oblongifolia,Garcinia oligantha,Cratoxylum cochinchinense andCalophyllum membranaceum were 0.285%, 0.199%, 0.857%, 0.161% and 0.006%, respectively. Isogarcinol was not detected inCratoxylum formosum orCalophyllum inophyllum. It was concluded that the method was convenient, accurate with high sensitivity, good stability and repeatability. It can be used for determination of isogarcinol content in Chinese herbal medicine.

AIM: To investigate the details of CD4 + T cell polarized to Th1/Th2 in vitro . METHODS: After isolated the PBMCs and blood-plasma from adult human peripheral blood by Ficoll-Hypaque centrifugation, the PBMC culture procedure with or without the self-blood -plasma was applied to polarize T cells in vitro , these cells were polarized by PHA(20 mg/L),non-PHA respectively. The polarized rates of Th cell after 24 h,48 h,72 h were estimated respectively by flow cytometry following two-color immunofluorescent staining. RESULTS: CD4 +T cell would polarize to Th1/Th2 two subsets after self-cytokines and PHA activation in vitro . The polarized rates of T cell after cultured for 24 h,48 h and 72 h were (13.28%?1.59%)/(12.70%?1.65%),(17.19%?1.03%)/(17.50%?1.30%),(19.49%?2.87%)/(18.58%?1.49%) respectively, but the polarized rates of T cell were very low if without self-blood-plasma. The difference between them was significant. The ratios of Th1/Th2 cells were about 1. CONCLUSION: CD4 +T cell from adult human peripheral blood would polarize to Th1/Th2 two subsets in the presence of self-blood-plasma and PHA(20 mg/L) in vitro , and the cell number of Th1 and Th2 would be in balance.

AIM: Peripheral blood mononuclear cells (PBMC) were cultured in vitro to study the effect of gossypol, a polyphenolic antifertility agent, on the activation of normal human T cells. METHODS: Double fluorescent staining together with flow cytometry was adopted to analyze the influence of gossypol on expression of the early activation antigen CD69 on T-lymphocytes under stimulation of mitogen or phorbol ester. RESULTS: Analysis of T cell activation in vitro revealed that preincubation of PBMC with 100 ?mol/L gossypol could completely inhibit the expression of early activation marker CD69 on CD3 + T cells in response to 10 mg/L PHA, and block T cell activation by 10 -7 mol/L PDB as well. The suppression of CD69 expression was dose-dependent and IC 50 of gossypol on PDB and PHA were (35 7?2 9) ?mol/L and (32 8?1.5) ?mol/L( ?s ), respectively. Besides, gossypol had similar inhibitory effect on CD69 expression of CD3 - lymphocytes. However, it did not have any significant effect on T cell surface molecule CD3 down-regulation. CONCLUSION: Gossypol could inhibit T cell activation in vitro in response to polyclonal activators, both PHA and PDB, suggesting that its action site may be at PKC or its downstream and that gossypol possessed potential immuno-regulatory effect. [

AIM: To investigate the influences of protein kinase C(PKC) inhibitors on the expression of interleukin-2(IL-2) and interferon-?(IFN-?) by in vitro activated T-lymphocytes. METHODS: Double fluorescent staining together with flow cytometry was adopted to detect intracellular cytokines and to analyze the effects of H7 and gossypol on IL-2 and IFN-? expression levels of T-lymphocytes stimulated with phorbol ester (PDB)+ionomycin(I) in the presence of monensin.RESULTS: The expression rates of IL-2 and IFN-? of CD3 + T cells stimulated with PDB+I for 4 h were 16.64?2.04 and 25.81?3.53( ?s ), respectively, which were significantly higher than that of control (1.06?0.22 and 3.12?0.77)( P

Objective To investigate the status of cellular immunity from Th cell polarization in pa-tients with different courses of condyloma acuminatum(CA).Methods The isolated PBMC were polarized by PHA and self-plasma for72hours,then followed by two-color immunofluorescent staining with anti-CD4-PE and anti-CCR5-FITC,or with anti-CD4-FITC and anti-CCR3-PE.Finally the stained cells were analyzed by flow-cytometry.Results The percentages of Th1/Th2cells of the short-course group and long-course group were(25.82?2.22)%/(14.80?1.14)%and(12.20?1.37)%/(13.74?0.99)%,respectively;the differ-ences between normal control and two CA groups were significant(P

AIM: To confirm that CD4~+CD25~+ regulatory T cells don't have an instinctive defection in IL-2 secretion, and to have an insight into the maturation state of CD4~+CD25~+ T cells in cord blood. METHODS: CD4~+CD25~+ and CD4~+CD25~- T cells were purified from cord blood of term infants (CB) and adult peripheral blood (PB) by autoMACS, and stimulated with PDB plus ionomycin. After 45 hours of culture, cells were detected for expression of CD69 and CD25 by flow cytometry, and the supernatants were measured for 7 kinds of cytokines by Luminex. RESULTS: CD4~+CD25~+ T cells from both CB and PB proliferated comparably with CD4~+CD25~- T cells when stimulated with PDB plus ionomycin. After 45 hours of culture, however, the CD4~+CD25~+ T cells underwent a tendency of cell death. Expression of CD25 was further upregulated when CD25~+ cells were activated. Under stimulation of PDB plus ionomycin, both CD4~+CD25~+ and CD4~+CD25~- T cells in PB secreted high levels of IFN-?, IL-2 and TNF-?, with CD25~+ cells secreted much higher level of IL-5, IL-4 and IL-10 than those in CD25~- cells; CD4~+CD25~+ and CD4~+CD25~- T cells in CB also secreted high level of IL-2 and TNF-? but much lower level of IFN-? than those in PB, and no secretion of IL-5, IL-4 and IL-10 was observed. CONCLUSION: CD4~+CD25~+ regulatory T cells don't have an instinctive defection in IL-2 secretion, otherwise there may be a different TCR signaling pattern in CD4~+CD25~+ T cells from traditional T cells. The CD4~+CD25~+ T cells in cord blood have not fully matured in function.