The aim of this study was to examine the priming effect of sphingosine 1-phosphate (S1P) on fMLP-activated neutrophils, mainly to detect the neutrophil respiratory burst products, and to investigate the signaling pathway involved in S1P activity. Flow cytometry was used to evaluate the new isolated neutrophil; the superoxide anion output was detected indirectly by cytochrome C reduction in respiratory burst; the dihydro-rhodamine 123 was used to detect the intensity of respiratory burst; the signal transduction pathways of neutrophil respiratory burst were explored by Western blot. The results showed that after pretreated with S1P, the level of superoxide anion released by fMLP-activated neutrophils significantly increased; the Rhodamine 123 mean fluorescence intensity in S1P primed fMLP-activated neutrophils group was significantly higher than that in fMLP treatment group; PI3K and Akt proteins involved in the signal pathway of neutrophil respiratory burst. It is concluded that S1P is a new priming reagent, which primes respiratory burst of fMLP-activated neutrophils; this signal pathway may be that S1P interacts with its receptor, activates PI3K, then activates Akt-transmitting signals through NADPH oxidase, finally results in the respiratory burst.
Cells, Cultured ; Humans ; Lysophospholipids ; metabolism ; NADPH Oxidases ; metabolism ; Neutrophils ; metabolism ; physiology ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptors, Lysosphingolipid ; metabolism ; Respiratory Burst ; Signal Transduction ; Sphingosine ; analogs & derivatives ; metabolism ; Superoxides ; metabolism

OBJECTIVETo study source apportionment of atmospheric PM10 (particle matter ≤ 10 µm in aerodynamic diameter) and PM2.5 (particle matter ≤ 2.5 µm in aerodynamic diameter) in Beijing,Urumqi and Qingdao, China.

METHODSThe atmospheric particle samples of PM10 and PM2.5 collected from Beijing between May 17th and June 18th, 2005, from Urumqi between April 20th and June 1st, 2006 and from Qingdao between April 4th and May 15th, 2005, were detected to trace the source apportionment by factor analysis and enrichment factor methods.

RESULTSIn Beijing, the source apportionment results derived from factor analysis model for PM10 were construction dust and soil sand dust (contributing rate of variance at 45.35%), industry dust, coal-combusted smoke and vehicle emissions (contributing rate at 31.83%), and biomass burning dust (13.57%). The main pollution element was Pb, while the content (median (minimum value-maximum value)was 0.216 (0.040-0.795) µg/m(3)) . As for PM2.5, the sources were construction dust and soil sand dust (38.86%), industry dust, coal-combusted smoke and vehicle emissions (25.73%), biomass burning dust (13.10%) and burning oil dust (11.92%). The main pollution element was Zn (0.365(0.126-0.808) µg/m(3)).In Urumqi, source apportionment results for PM10 were soil sand dust and coal-combusted dust(49.75%), industry dust, vehicle emissions and secondary particles dust (30.65%). The main characteristic pollution element was Cd (0.463(0.033-1.351) ng/m(3)). As for PM2.5, the sources were soil sand dust and coal-combusted dust (43.26%), secondary particles dust (22.29%), industry dust and vehicle emissions (20.50%). The main characteristic pollution element was As (14.599 (1.696-36.741) µg/m(3)).In Qingdao, source apportionment results for PM10 were construction dust (30.91%), vehicle emissions and industry dust (29.65%) and secondary particles dust (28.99%). The main characteristic pollution element was Pb (64.071 (5.846-346.831) µg/m(3)). As for PM2.5, the sources were secondary particles dust, industry dust and vehicle emissions (49.82%) and construction dust (33.71%). The main characteristic pollution element was Pb(57.340 (5.004-241.559) µg/m(3)).Enrichment factors of Zn, Pb, As and Cd in PM2.5 were higher than those in PM10 both in Beijing and Urumqi.

CONCLUSIONThe major sources of the atmospheric particles PM10 and PM2.5 in Beijing were cement dust from construction sites and sand dust from soil; while the major sources of those in Urumqi were pollution by smoke and sand dust from burning coal. The major sources of the atmospheric particles PM10 in Qingdao were cement dust from construction sites; however, the major sources of PM2.5 there were secondary particles dust, industry dust and vehicle emissions. According to our study, the heavy metal elements were likely to gather in PM2.5.

Air Pollutants ; analysis ; China ; Cities ; Dust ; analysis ; Environmental Monitoring ; methods ; Particle Size ; Seasons ; Vehicle Emissions ; analysis

OBJECTIVETo study the stereoselective glucuronidation of carvedilol (CARV) by three Chinese liver microsomes.

METHODSThe metabolites of CARV were identified by a hydrolysis reaction with beta-glucuronidase and HPLC-MS/MS. The enzyme kinetics for CARV enantiomers glucuronidation was determined by a reversed phase-high pressure liquid chromatography (RP-HPLC) assay using (S)-propafenone as internal standard after precolumn derivatization with 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosylisothiocyanate.

RESULTSTwo CARV glucuronides were found in three Chinese liver microsomes incubated with CARV. The non-linear regression analysis showed that the values of K(m) and V(max) for (S)-CARV and (R)-CARV enantiomers were (118+/-44) micromol/L, (2 500+/-833) pmol/(min.mg protein) and (24+/-7) micromol/L, (953+/-399) pmol/(min.mg protein), respectively.

CONCLUSIONThese results suggested that there was a significant (P<0.05) stereoselective glucuronidation of CARV enantiomers in three Chinese liver microsomes, which might partly explain the enantioselective pharmacokinetics of CARV.

Carbazoles ; metabolism ; China ; Glucuronic Acid ; metabolism ; Glucuronides ; metabolism ; Microsomes, Liver ; metabolism ; Propanolamines ; metabolism ; Stereoisomerism

OBJECTIVETo study the influence of human plasma exosomes-like vesicles on the regulatory function of macrophages.

METHODSThe exosomes-like vesicles were purified from healthy donors plasma with a series of high-speed centrifugation and ultrafiltration. Macrophages were derived from cultured human blood monocytes. The molecular markers of macrophages were assayed by FACS. After cultured with exosomes-like vesicles, the changes of macrophages cytoplasma Ca(2+), and related genes and proteins were assayed by FACS, RT-PCR and Western Blot, respectively.

RESULTSAfter cultured with exosomes-like vesicles, mean fluorescent intensity (MFI) of macrophages cytoplasma Ca(2+) was increased. The vesicles enhanced macrophages to express cytokines genes, the expression of IL-1β and TNF-α genes being increased by 0.85 and 1.69 times respectively at 2 h, and that of IL-6 gene 3.7 times compared with the control at 8 h. However, the vesicles inhibited the expression of macrophages IL-10 gene, had no influence on the Frizzled5 receptor expression and could induce CaMKII phosphorylation.

CONCLUSIONSExosomes-like vesicles can up-regulat macrophages expression of inflammatory cytokines genes, and increase the secretion of inflammatory cytokines by activating the Wnt5A-Ca(2+) signaling pathway.

Adolescent ; Adult ; Calcium ; metabolism ; Calcium Signaling ; Exosomes ; Female ; Humans ; Macrophage Activation ; Macrophages ; metabolism ; Middle Aged ; Proto-Oncogene Proteins ; metabolism ; Wnt Proteins ; metabolism ; Wnt-5a Protein ; Young Adult

OBJECTIVETo investigate the effects of murine bone marrow endothelial cell conditioned medium (mBMEC-CM) on the growth of yolk sac hematopoietic progenitors.

METHODSThe serum-free mBMEC-CM was obtained from subcultures of murine endothelial cell line derived from bone marrow which was established in our laboratory. The murine yolk sacs were harvested on day 8.5 postcoitus (pc) and incubated with 0.1% collagenase in 10% fetal calf serum at 37 degrees C for 40 minutes. Yolk sac cells were incubated in tissue culture dishes at 37 degrees C for 1 hour. Nonadherent cells were collected for semisolid culture assay of granulocyte-macrophage colony forming unit (CFU-GM) and high proliferative potential-colony forming cell (HPP-CFC) after being cultured in DMEM with 10% mBMEC-CM and 10% FBS for 24 hours. The number of CFU-GM and HPP-CFC was counted at day 7 and day 14 respectively.

RESULTSThe growth of CFU-GM and HPP-CFC was supported by mBMEC-CM with GM-CSF. mBMEC-CM could induce the proliferation and differentiation of yolk sac hematopoietic stem cells and progenitors in liquid culture system. The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (119.5 +/- 5.7)% and (130.8 +/- 9.8)% respectively after 24 hours liquid culture (P < 0.05). The expansion effects of mBMEC-CM on CFU-GM and HPP-CFC were enhanced by compounded with flt3 ligand (FL) and thrombopoietin (TPO). The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (132.0 +/- 6.2)% and (176.9 +/- 12.8)% respectively after 24 hours liquid culture (P < 0.01).

CONCLUSIONMurine bone marrow endothelial cell conditioned medium could support the growth and proliferation of yolk sac hematopoitic stem cells and progenitors, and this promoting effect was further enhanced by addition of FL and TPO.

Animals ; Bone Marrow Cells ; cytology ; Cell Division ; Cells, Cultured ; Culture Media, Serum-Free ; Endothelium ; cytology ; Female ; Hematopoiesis ; Hematopoietic Stem Cells ; cytology ; Male ; Mice ; Yolk Sac ; cytology

OBJECTIVETo explore the biological characteristic of third-party-derived tolerogenic DC(tDC) and the influence of third-party-derived tDC on acute graft-versus-host-disease (aGVHD) following allogeneic bone marrow transplantation (allo-BMT) in mice.

METHODStDC from bone marrow cells of D1 mice was cultured with low doses of GM-CSF, IL-10 and TGF-β1D1. The phenotype, expression of cytokines and function associated molecules were identified with FACS and RT-PCR. Mixed lymphocyte reaction was applied to analyze the influence of third-party-derived tDC on allo-CD4(+)T cells proliferation in vitro. Different doses of D1-tDC were adoptive transferred in the aGVHD model in allogeneic BMT which B6 mice as donors and D2 mice as recipients. Survival time, clinical GVHD score and the levels of Th1/2 cytokines in serum were monitored after allo-BMT using the aGVHD model as control.

RESULTStDC expressed lower levels of MHC II and co-stimulatory molecules, such as CD80, CD86 and CD40, even when stimulated by LPS. The results by RT-PCR indicated that tDC expressed low levels of IL-12p40 and high levels of immunosuppressive molecules, such as IL-10, TGF-β, Fas Ligand, indoleamine 2, 3-dioxygenase (IDO) and arginase. In the allogeneic MLR, third-party tDC suppressed allo-CD4(+)T cells proliferation, which was relative to the dose of tDC. In the B6→D2 mouse model, all aGVHD mice died within 18 days. Remarkably, if 10(4) third-party tDC were transferred, 60% mice survived at least 60 days. When the doses of tDC were reduced to 10(3) cells, only 20% of mice survived day 60, and when increased tDC to 10(5), all of the mice died within day 37 after allo-BMT. The cytokine levels in serum indicated that 10(4) tDC-treated mice secreted in vivo high level of IL-10 21d after BMT (P < 0.05), the levels of IL-10 in 10(3), 10(4) and 10(5) tDC-treated mice were (114.23 ± 7.78), (646.18 ± 212.02), (121.97 ± 10.47) ng/L, respectively.

CONCLUSIONThird-party tDC could suppress allo-CD4(+)T cells proliferation in vitro and prevent aGVHD in allogeneic BMT mode, which may be mediated by modulating tolerogenic cytokines secretion, such as IL-10. And this effect was associated with the dose of tDC. Adoptive therapy by transfusing third-party tDC cultured with low doses of GM-CSF, IL-10 and TGF-β1 could significantly prolong the survival of recipients and prevent aGVHD in allogeneic BMT.

Animals ; Bone Marrow Transplantation ; adverse effects ; CD4-Positive T-Lymphocytes ; cytology ; Cell Proliferation ; Dendritic Cells ; cytology ; immunology ; metabolism ; Graft vs Host Disease ; prevention & control ; Interleukin-10 ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Transforming Growth Factor beta1 ; immunology ; Transplantation, Homologous

OBJECTIVE: To study the value of anti-neutrophil cytoplasmic antibody (ANCA) in assessing the severity of bronchiolitis obliterans (BO) in children. METHODS: A prospective analysis was performed on 59 children who were diagnosed with BO from June 2009 to October 2014. ELISA was used to measure the concentrations of myeloperoxidase (MPO)-ANCA and proteinase 3 (PR3)-ANCA in serum. According to the results of ELISA, the children were divided into three groups: double-negative ANCA (n=22), single-positive ANCA (n=17), and double-positive ANCA (n=20). The three groups were compared in terms of the scores of BO risk factors, clinical symptoms, chest high-resolution computed tomography (HRCT), and lung pathology on admission, as well as the changes in the expression level of ANCA and the scores of clinical symptoms and chest HRCT over time. RESULTS: Compared with the double-negative ANCA group, the double-positive ANCA group had a significantly higher score of BO risk factors (P<0.05), and the single-positive ANCA group and the double-positive ANCA group had significantly higher scores of clinical symptoms, chest HRCT, and lung pathology (P<0.05). The children were followed up for 6 months after discharge, and there were significant reductions in MPO-ANCA and PR3-ANCA titers from admission and discharge to the end of follow-up (P<0.05), as well as a significant reduction in the score of clinical symptoms from admission to the end of follow-up (P<0.05), while there was no significant change in the score of chest HRCT from admission to the end of follow-up (P>0.05). The single-positive ANCA and double-positive ANCA groups still had a significantly higher score of clinical symptoms than the double-negative ANCA group (P<0.05). CONCLUSIONS The expression level of ANCA is correlated with the severity of BO in children and thus has certain clinical significance in disease evaluation.
Antibodies, Antineutrophil Cytoplasmic ; Bronchiolitis Obliterans ; Child ; Humans ; Myeloblastin ; Peroxidase ; Prospective Studies

Objective To analyze the efficacy of ponatinib as salvage therapy in relapse chronic myeloid leukemia with T315I mutation (CML-T315D after allogeneic stem cell transplantation (allo-HSCT).Methods Twelve patients with CML-T315I (10 cases of T315I mutation before transplantation and 2 cases of T315I mutation at the time of relapse after transplantation) were included in this retrospective analysis.Ponatinib was used as single agent or combined with chemotherapy and/or donor lymphocyte infusion.The samples obtained for RTQ-PCR were also analyzed for the BCR ABL1 mutation by direct sequencing.Scanning of the ABL KD (amino acids 219-506) for the presence of mutations was sequenced by Sanger.Results In 12 patients with relapse after transplantation,2 patients with molecular relapse were treated with only single-agent ponatinib,and among 10 patients with hematologic relapse,1 patient was treated with single-agent ponatinib and 3 patients were given ponatinib combined with donor lymphocyte infusion (DLI),the remaining 6 patients were treated with ponatinib combined with chemotherapy and DLI.After the treatment with ponatinib,11 patients had a good response,10 patients obtained complete hematologic remission (CHR),1 patient obtained partial hematologic remission (PHR) and 1 patient had no response (NR).For cytogenetic response,10 patients obtained complete cytogenetic response (CCyR),1 patient obtained partial cytogenetic response (PCyR) and one patient had no cytogenetic response.For the molecular biological response,9 patients obtained complete molecular response (CMR),1 patient obtained majore molecular response (MMR) and 2 patients had no molecular biological response.The median time to obtain CHR was 36 days (29-96 days),the median time to obtain CCyR was 63 days (32-127 days),and the median time to obtain CMR was 89 days (27-152 days).The median follow-up time after treatment with ponatinib was 598 (range,93-1470) days,9 patients survived and 3 died.Causes of deaths included leukemia relapse (n =2)and ineffective treatment (n =1).The 2-year overall and disease-free survival rate after relapse in 12 patients was 75.0％ ± 12.5％ and 31.7％ ± 14.9％,respectively.Conclusion This small sample data suggested that ponatinib as salvage therapy had a good response to the relapse CML-T315I after allo-HSCT.

AIM: To observe the anti-oxidative activity and adverse laxative effect of raw, traditional processed and fermented products of Polygoni Multiflori Radix (PMR), and furthermore, to evaluate the fermentation method used in the processing procedure of PMR. METHODS: In vitro ferric reducing antioxidant power (FRAP) assay was carried out to evaluate the anti-oxidative activity. Modulation of normal defecation and effect on gastrointestinal motility in mice were carried out to investigate their adverse laxative effect. RESULTS: Fermented PMR induced less severe laxative adverse effect than Polygoni Multiflori Radix Praeparata (PMRP). PMR fermented with Rhizopus sp. (FB) could modulate the defecation significantly. The gastrointestinal motility was inhibited by PMRP and PMR fermented with Rhizopus oryzae (FA). FA and FB showed better antioxidant activity than PMRP in 50% and 95% ethanol group. Contents of 2, 3, 5, 4'-tetrahydroxy-stilbene-2-O-β-D-glucoside (TSG) were reduced significantly after traditional processing but maintained after fermentation. Emodin and physcion were increased after traditional processing and fermented with Rhizopus oryzae. CONCLUSION All processing procedure, including fermentation, might reduce its anti-oxidative activity. However, most of the processed products could lessen the adverse effect on gastrointestinal tract compared to PMR. Fermentation with Rhizopus oryzae was considered as a promising processing method of PMR.
Animals ; Antioxidants ; pharmacology ; Defecation ; drug effects ; Emodin ; adverse effects ; analogs & derivatives ; pharmacology ; Female ; Fermentation ; Gastrointestinal Motility ; drug effects ; Gastrointestinal Tract ; drug effects ; Laxatives ; adverse effects ; Male ; Mice ; Mice, Inbred Strains ; Plant Extracts ; adverse effects ; pharmacology ; Plant Roots ; chemistry ; Polygonum ; adverse effects ; chemistry ; Rhizopus

Objective:To observe the effect of compressive low elastic bandage on upper limb lymphedema after breast cancer surgery. Methods:From November, 2017 to December, 2018, 64 patients with upper limb lymphedema after breast cancer surgery were randomly divided into control group (n = 32) and observation group (n = 32). Both groups accepted manual lymphatic drainage and low elastic bandage, while the observation group increased the pressure with sponge pads on the fibrous swelling site as bandaging, five times a week for two weeks. They were measured L-Dex with bioelectrical spectroscopy before the first treatment (T1), before the second treatment (T2), before the third treatment (T3), before the sixth treatment (T4), and 24 hours after the tenth treatment (T5), respectively. The arm circumference of affected side and un-affected side was also measured at T1 and T5. Results:L-Dex decreased since T2 in both groups, but increased at T4 in the observation group and more than that of the control group (t = 2.13, P < 0.05), and less than that of the control group at T5 (t = -2.29, P < 0.05). The difference of arm circumference between affected side and un-affected side was less in the observation group than in the control group (t = -3.78, P < 0.001)。 Conclusion:Low elastic bandage combined with lymphatic drainage technique can alleviate the fibrous swelling. On the basis of low elastic bandage with local sponge pad compression is more effective than simple low elastic bandage on fibrous swelling after lymphedema of upper limbs after breast cancer surgery.