Association of polycyclic aromatic hydrocarbon exposure and CYP1A1 gene and enzyme activity of peripheral blood monocyte cells in coke oven workers / 中华劳动卫生职业病杂志

Objective@#To explore the association between polycyclic aromatic hydrocarbons (PAHs) exposure and cytochrome P450 CYP1A1 expression at gene and enzyme activity levels in the peripheral blood monocyte cells in coke oven workers, and to provide a certain basis for the biological monitoring of health damage in coke oven workers.@*Methods@#We surveyed 118 coke oven workers and 63 controls (energy power workers in the same company) using self-designed questionnaire, determined their post-shift urinary 1-hydroxypyrene (1-OH-Py) concentration using high performance liquid chromatography (HPLC)-fluorescence detector method. We also isolated the peripheral blood mononuclear cell (PBMC) from fasting venous blood, and detected DNA damage using comet assay, CYP1A1 mRNA level using the real-time fluorescent quantitative PCR (FQ-PCR), and EROD activity using spectrophotometry. Statistical analyses including one-way analysis of variance and multiple linear regressions were used to analyze the association of urinary 1-OH-Py and CYP1A1 mRNA level and EROD activity.@*Results@#Compared to the control group, the urinary 1-OH-Py concentration and PBMC DNA tail moment were significantly increased in coke oven workers (P<0.05), and CYP1A1 gene level and EROD activity in PBMC were significantly decreased (P<0.05). Multiple linear regression showed that a ten-fold increase of urinary 1-OH-Pycon centration was associated with a decrease of 0.77 (95%CI: -1.33--0.21) in CYP1A1 gene level, and a decline of 0.15 (95%CI: -0.76--0.16) in EROD activity of PBMC in coke oven workers (P<0.05).@*Conclusion@#Occupational PAHs exposure induced DNA damage, which was associated with the decreased level in CYP1A1 gene cavel and EROD activity in PBMC of coke oven workers.

Peripheral blood absolute lymphocyte count, absolute monocyte count and their ratio in prognosis prediction of newly diagnosed multiple myeloma / 肿瘤

Objective: To investigate the value of peripheral blood absolute lymphocyte count (ALC), absolute monocyte count (AMC) and ALC/AMC ratio (LMR) in predicting the prognosis of patients with newly diagnosed multiple myeloma (MM). Methods: Retrospective analysis was performed in 190 patients with newly diagnosed MM from Tianjin Medical University Cancer Institute and Hospital between January 2005 and December 201 5. The relationships of peripheral blood ALC, AMC and LMR with peripheral blood hemoglobin (Hb), β2-microglobulin (β2-MG) and lactate dehydrogenase (LDH) and the proportion of bone marrow plasma cells were analyzed. The cutoff values of ALC, AMC and LMR were determined by the receiver operating characteristic (ROC) curve in patients with newly diagnosed MM. Survival analysis was performed using Kaplan-Meier method and the log-rank test for univariate analysis of prognosis, and a COX proportional hazard model was used for multivariate analysis of prognosis. Results: The cutoff values of ALC, AMC and LMR determined by ROC curve were 1.24×109/L, 0.60×109/L and 3.90, respectively. According to these cutoff values, the patients were divided into high value groups and low value groups. The results of multivariate analysis showed that ALC 247 U/L (HR: 1.972, 95% CI: 1.087-3.576; P = 0.025) were independent poor prognostic factors in untreated MM patients. According to the number of poor prognostic factors (each poor prognostic factor was scored as one), the patients were divided into score 0, 1-2 and 3 groups, the overall survival and progression-free survival of the three groups were significantly different (all P < 0.05). Conclusion: Lower ALC and LMR values may indicate poor prognosis. ALC < 1.24×109/L and LMC≤3.90 maybe the independent poor prognostic factors in patients with newly diagnosed MM.

The expression of TLR4 on peripheral blood monocytes from uremic patients with diabetic nephropathy and its relation with plasma MCP-1 concentration / 中国免疫学杂志

Objective:To explore the expression of TLR4 on peripheral blood monocytes in uremic patients with diabetic nephropathy and the relation between TLR4 and diabetic nephropathy.Methods:RT-PCR analysis and flow cytometry were undertaken to measure the expression of TLR4 mRNA and protein.The plasma concentration of MCP-1 was measured with ELISA.Results:The gene and protein expression of TLR4 on peripheral blood monocytes in patients with diabetic nephropathy was significantly increased compared with that of the health control,and TLR4 expression was positively related with the the plasma concentration of MCP-1.Conclusion:TLR4 expression is upregulated on peripheral blood monocytes in patients with diabetic nephropathy and the upregulation is related with the pathogenesis of diabetic nephropathy.

Degeneration of Dendritic Cells from Human Peripheral Blood and Acquirement Antigen from Apoptotic Hepatochlangioma Cells / 中国肿瘤临床

Objective:To eastablish the efficient presentation of antigen from apoptotic cells by human DC from peripheral blood. Methods: using recombinant human granulocyte/macrophage colonystimulating factor(GM- CSF) and interleukin 4 (IL- 4 ) we have established dendritic cells (DC)from peripheral blood monocyte that maintain the antigen capturing and processing capacity characteristic of immature dendritic cells in vivo. GM - CSF 50ng/ml , IL- 41 000ng/ml once two days(total four). on the 3 rd day of culture, immature DC and apoptotic hepatochlangioma cells were in coculture lasting 7 days. Results:these cells had typical dendritic morphology, express high levels of CD1a ,B7 and acquired antigen from apoptotic cells and induced an increased T cell stimulatory capacity in MLR. Conclusions:we have established DC from blood mononuclear tells using GM- CSF and IL- 4 and DC can be efficiently drived from apoptotic cells and can induce the increase of T cells obviously. It probably becomes an effective approach of antigen transduced with DC.

Clinical implication of serum TNF-alpha and IL-1beta measurement in patients with sepsis / 결핵

BACKGROUND: It is well known that when macrophages are stimulated with endotoxin, they produce a wide variety of cytokine mediators, including TNF-α and IL-1β. However, there is an alterationnin the macrophages responsiveness when they are challenged with repeated bouts of endotoxin, termed 'endotoxin tolerance' which is regarded as a self-protective phenomenon from continuous stimulation. In this study, endotoxin tolerance in the peripheral blood monocytes of sepsis patients was evaluated. METHODS: Fourteen patients with organism-documented sepsis were included. The severity of illness was evaluated by APACHE IIscore. Peripheral blood monocytes were isolated from the patients and diluted to 1×105/well. After stimulation with endotoxin(LPS of E. coli O114:B4, 100 ng/ml), they were incubated at 37℃ in 5% CO2 incubator for 24 hours. Supernatant was collected for the measurement of TNF-αand IL-1β with ELISA method. Peripheral blood monocytes of seven healthy volunteers were used as control. RESULTS: The APACHE IIscore(mean±SD) of the patients at the time of blood sampling was 12.2±5.7. The primary infection foci were urinary tract infection, pneumonia, subacute bacterial endocarditis, and catheter related infection, etc. The causative organisms were gram negative rods(10 cases), gram positive cocci(6 cases) with two cases of mixed infection. Serum TNF-α could be measured in 4 cases with 29.9±27.7 pg/ml. Serum IL-1β was measureable in only one patient. The TNF-α level of supernatant of cultured peripheral blood monocytes was 2,703±2,066 pg/ml in patients and 2,102±1,914 pg/ml in controls. The IL-1β level of supernatant was 884±1,050 pg/ml in patients and 575±558 pg/ml in controls. There was no difference of TNF-α and IL-1β level between patients and controls. CONCLUSION: We cannot prove the phenomenon of endotoxin tolerance in this study. Future study needs to be focused on the more severe sepsis patients who were taken for sampling earlier. Addition of serum to the culture medium could be an another valuable option for the success of this study.

Expression and significance of toll-like receptor 4 by peripheral blood mononuclear cell in rats with adjuvant-induced arthritis / 西安交通大学学报(医学版)

Objective To demonstrate the role of innate immunity reaction in rheumatoid arthritis and other autoimmune diseases by studying the expression of toll-like receptor 4(TLR4) by peripheral blood mononuclear cell(PBMC) in rats with adjuvant-induced arthritis(AIA) and analyzing the relationship between expression of TLR4 by PBMC and score of arthritis severity.Methods Both the proportion of CD14+TLR4+ cells in CD14+ cells and the mean intensity of fluorescence(MIF) of TLR4+ cells were analyzed by flow cytometry.Expression of TLR4 mRNA by PBMC was determined by Northern blot.CD14+TLR4+/CD14+ ratio,TLR4 MIF and expression of TLR4 mRNA by PBMC in the model group at 15 days after AIA were compared,respectively,with normal control group and model group at 25 days.The correlation between TLR4 MIF and score of arthritis severity was analyzed by Mann-Whitey U test.Results CD14+TLR4+/CD14+ ratio,TLR4 MIF and TLR4 mRNA expression in model group at 15 days after AIA were obviously increased compared with those in control and model groups at 25 days(P0.05).It indicated that TLR4 participated in the pathogenesis of rheumatoid arthritis.Conclusion TLR4 expressed by PBMC is closely associated with pathogenesis and pathophysiology of rheumatoid arthritis,which suggests that innate immunity response plays an important role in pathogenesis of RA and other autoimmune diseases.