To evaluate the effect of bFGF on the calcium influx in SGCs and its antagonistical effect on streptomycin. The SGCs of guinea pig were isolated using an enzyme machine methods and loaded with 10?mol/L Fluo 3/AM for 30 min at 37℃,then cultured 60min at room temperature. Individual Fluo 3 loaded SGCs were examined with a confocal microscope (ACAS Ultima, USA) using a 20 x objective lens and linear scan mean.The level of SGCs[Ca 2+ ] i was steady within the process of normal extracellular liquid perfusion. SGCs[Ca 2+ ] i was increased when SGCs were perfused with 150mmol/L high potassium media(10/10) and normal media containing 1nmol/L bFGF(8/9), but the level of SGCs[Ca 2+ ] i did not change in high potassium free calcium media(9/9) and free calcium media containing 1nmol/L bFGF(10/11). After the treatment with 1 mmol/L streptomycin, the level of SGCs[Ca 2+ ] i was increased 1/11 and 5/12, respectively, when perfused high potassium media and bFGF media. And after being treated with 1nmol/L bFGF, high potassium media, SGCs[Ca 2+ ] i was increased more obviously and persisted for a longer time. After being treated with 1 mmol/L streptomycin, 0 1nmol/L, 1nmol/L and 10nmol/L bFGF, respectively, and 14/14 the level of SGCs[Ca 2+ ] i was increased 5/11, 9/12, and 14/14 when perfused high potassium media. High potassium media and bFGF perfusion could result in an increase of SGCs[Ca 2+ ] i ,and SGCs[Ca 2+ ] i increase was the result of calcium influx, and there was a synergic effect between high potassium and bFGF. Streptomycin could block the process of calcium influx induced by high potassium media, the blocking effect could be antagonized by bFGF, and the antagonistic effect was bFGF concentration dependent.

OBJECTIVETo investigate the expression of CD160 on the surface of human natural killer (NK) cells and its possible relationship with hematological malignancies.

METHODSCD160 expression on human leukemia cell line NK92 cells was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The proliferation characteristics and cell surface markers of this cell line were determined. Cytotoxicity of NK92 against 2 human myeloid leukemia cell lines, K562 and THP-1 was analyzed ex vivo. CD160 blocking antibody CL1-R2 was employed to clarify its role in NK cell mediated cytolysis. Then, the expression of CD160 on NK cells in peripheral blood from various patients with hematological malignancies were measured by flow cytometry.

RESULTSThe mRNA and protein levels of CD160 expressions on NK92 cells were confirmed by RT-PCR and Western blot, respectively. The flow cytometry results demonstrated that the strong positive expression of CD160 could be detected on the NK92 cell surface. NK92 could effectively kill K562 and THP-1 cells, while the cytolysis effect was abrogated in the presence of CD160 blocking antibody CL1-R2. The high levels of HVEM were expressed on both target cells, but the HLA class I molecules were absent on K562. The expression of CD160 on CD3CD56 NK cells in peripheral blood from patients with acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL) and allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients was significant lower than that in the normal controls (P<0.05).

CONCLUSIONThe cytolysis function of human NK cells is mediated partially by CD160 molecule. The decrease of CD160 expression on NK cells from patients with various hematological malignancies implies that down-regulation of CD160 expression may be a novel mechanism of tumor immune escape.

OBJECTIVE: To investigate the characteristic changes of the plasma cytokine profile in Chinese patients with idiopathic multicentric Castleman diseases (iMCD). METHODS: The plasma samples from 22 patients with confirmed diagnosis of iMCD were collected before treatments; Specimens from 17 patients with newly diagnosed multiple myeloma, 10 non Hodgkin's lymphoma, and 15 healthy donors were used as control. Seventeen kinds of cytokines were measured by cytokine beads array (CBA) and ELISA respectively. RESULTS: Six cytokines were measured by ELISA. The concentrations of IL-2, IL-6, IL-21 and VEGF were significantly higher in the plasma of iMCD patients than those of the healthy donors (P＜0.01) and the level of IL-21 was highest in the iMCD group. There was no significant difference in the levels of IL-1β and IL-4 between the iMCD and healthy donor groups. Thirteen cytokines were measured by CBA assay, besides IL-6 level was confirmed to be higher in iMCD group than that in healthy controls (P＜0.01）, IL-12-p70 and IL-33 levels were also higher in iMCD group than those in control group (P＜0.05）, no significant difference of the rest cytokines was found between iMCD and the control group. CONCLUSION IL-6 and VEGF has shown to involved in the pathogenesis of iMCD, the results of preliminary study imply the role of IL-2 、IL-21、IL-12-p70 and IL-33 in this rare lymphoproliferative disease. Further studies are needed to elucidate the mechanism of these cytokines, which may shed some light on the identification of novel therapeutic targets against iMCD.
Castleman Disease ; Cytokines ; Humans ; Interleukin-12 ; Interleukin-1beta ; Plasma