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Objective:To investigate the expression of macrophage migration inhibitory factor (MIF) in bone marrow fluid and peripheral blood of patients with acute myeloid leukemia (AML) and its effect on the expression of interleukin-8 (IL-8) in bone marrow mesenchymal stem cells (BM-MSC).Methods:Fifty bone marrow fluid samples and 50 peripheral blood samples were collected from 50 patients with AML diagnosed in the First People's Hospital of Yunnan Province from October 2017 to January 2019, of which 17 patients were newly diagnosed, 26 patients were complete remission (CR), and 7 patients were partial remission (PR) or non-remission (NR). Fifty plasma samples from 50 healthy subjects and 50 bone marrow fluid samples from 50 patients with iron deficiency anemia were used as the controls. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of MIF protein in the samples, and the relationship between MIF expression level and clinicopathological characteristics of AML patients was analyzed. BM-MSC was successfully isolated and cultured from 42 bone marrow fluid samples of AML patients, the suitable samples for experiment were chosen and divided into BM-MSC control group (untreated BM-MSC), recombinant human macrophage migration inhibitory factor (rhMIF) group and rhMIF+ISO-1 group. ELISA and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression level of IL-8 protein and mRNA in each BM-MSC group.Results:The expression levels of MIF protein in bone marrow fluid and plasma in AML group were (24.9±7.7) ng/ml and (60.5±12.1) ng/ml, the difference was statistically significant ( P < 0.01), and those in control group were (5.3±2.6) ng/ml and (2.0±1.1) ng/ml, respectively, and there were statistical differences between the two groups (t values were 136.71, 33.97 and 17.58, all P < 0.01). MIF protein expression levels in bone marrow fluid and plasma of AML patients in newly diagnosed group and PR+NR group were higher than those in CR group, and the differences were statistically significant (all P < 0.01). MIF protein expression levels were higher in bone marrow fluid and plasma of patients with ≥60 years of age, peripheral blood white blood cell count ≥30×10 9/L and bone marrow myeloblast ratio > 0.50 (all P < 0.05), but the differences were not statistically significant between patients with different gender (both P > 0.05). The detection results of each BM-MSC group showed that rhMIF promoted the IL-8 expression in BM-MSC at the gene and protein levels, which could be inhibited by the MIF inhibitor ISO-1 (all P < 0.01). Conclusion:The increased expression levels of MIF in bone marrow fluid and plasma of patients with AML are associated with the disease progression, and rhMIF can promote the expression of IL-8 in BM-MSC.
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Hepatocellular carcinoma (HCC) is the second largest cause of cancer deaths worldwide.TLRs signal transduction pathways are related to the progress of HCC.As an innate immune recognition receptors on the surface of the cell,TLRs are involved in the ligand-mediated activation of various cells and the secretion of cytokines in liver through signal transduction,which is part of the cause and development of HCC.A better understanding of TLRs and these signaling pathways in the liver will help us understand the mechanism of hepatocarcinogenesis and provide a new therapeutic target for HCC.
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Objective to explore the correlation between interferon-γ receptor(IFNGR)2 amino acid sites Val14Met and GIn64Arg polymorphism and atherosclerosis plaque stability in Yunnan Han nationality.Methods The patients with unstable atherosclerotic plaque in our hospital from March 2014 to March 2015 were collected as the observation group and contemporaneous patients with stable atherosclerotic plaque/non-plaque as the control group.The peripheral venous blood was collected for extracting genomic DNA.IFNGR Va114Met and GIn64Arg loci genotype was detected by the PCR product direct sequencing method.The sequencing results were analyzed by adopting the DNAStar and GeneTool software.The levels of plasma cytokines(IFN-γ)was detected by the flow cytometry.Results Two hundreds and four native Han patients were included in this study,including the observation group,109 cases,age(76.89±12.08)years old;the control group,95 cases,age(65.99±16.32)years old.The polymorphism change of IFNGR1 Vall4Met loci was not found irn the two groups.In the observation group,the frequency of IFNGR2 Gln64Arg genotype AA was 51.95％(40/77),which of AGwas 53.06 ％(52/98)and which of GG was 58.62％(17/29);in the control group,the frequencies were 48.05 ％ (37/77),46.94 ％ (46/98) and 41.38 ％ (12/29),chi-square test,P =0.824.The IFNGR2 Gln64Arg genotypes AA,AG and GG had no relation with atherosclerotic plaque stability.The A and G allele frequencies in the observation group were 52.38％ (132/252)and 55.13 ％ (86/156)respectively,which of the control group were 47.62 ％ (120/252)and 44.87％ (70/156),chi-square test's P=0.661.The IFNGR2 Gln64Arg A/G allele had no relation with atherosclerotic plaque stability.By Hardy-Weinberg genetic balance test,the gene frequency in this sample population was in accordance with the genetic equilibrium law.The plasma IFN-γ level in the observation group was(4.60 ± 1.91)ng/mL,which in the control group was (4.88 ± 2.10)ng/ mL,the difference was not statistically significant(P=0.318);the plasma IFN-γ content had no relation with atherosclerotic plaque stability(P=0.308).Conclusion The genetic polymorphisms of IFNGR can not serve as a warning indicator of atherosclerotic plaque stability.
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[ ABSTRACT] Tyrosine kinase inhibitors ( TKIs) are now advocated as the first-line treatment for chronic myeloid leukemia ( CML) , but facing resistance and relapse .Leukemia stem cells ( LSCs ) are leukemia-initiating cells as the source of resistance and relapse .It is therefore important to discover the molecular biomarker of LSCs for developing anti -LSC strategies in leukemic therapy .15-Lipoxygenase (15-LO) is a key enzyme in the pathway of arachidonic acid and plays an important role in the occurrence and development of CML , which is specifically required for chronic myeloid LSCs . This review summarizes the influence of 15-LO on the chronic myeloid LSC characteristics of marked survival , self-renewal, proliferation , differentiation and apoptosis .
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Objective To investigate the changes of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpressions in bone marrow of chro‐nic myeloid leukemia(CML)patients .Methods The IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γexpression levels were detected by u‐sing flow cytometry in 30 cases of CML chronic phase(CML‐CP) ,21 cases of CML accelerated phase(CML‐AP) ,15 cases of CML blastic phase(CML‐BP) ,42 cases of CML remission after treatment and 7 cases of non‐remission .Then the detection results were compared with those in the control group .Results The expression levels of INF‐γ and IL‐2 in each CML groups were lower than those in the control group(PCML‐AP> CML‐CP ,the difference among groups had statistical significance (P0 .05) .Conclusion The changes of serum cytokines in bone marrow microenvironment of CMLpatients have certain significance to the occurrence ,development and prognosis of CML ;the de‐tection of IL‐1β,IL‐2 ,IL‐4 ,IL‐6 ,IL‐10 and INF‐γlevels in bone marrow is hopeful to provide new ideas and theoretical basis for im‐mune therapy and prognosis judgment of CML patients .
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Objective To investigate the inhibitory effect of rhIFN-γ on human chronic myeloid leukemia cell line K562,and the impaction on the expression of CD123.Methods MTT method was used to test cell relative viability with rhIFN-γ at different concentrations.The expression of CD123 on K562 ceils was detected by flow cytometry.Results The relative inhibition of K562 cells proliferation was hampered when the cells were treated with rhIFN-γ for 72 h at the concentration of 5 x 10P ～ 108 U/L,respectively.However,rhIFN-γ at 2 x 105 U/L was benefit to K562 cells proliferation.After treatment with rhIFN-γ at 0,2 x 105 U/L and 107 U/L,the percentages of CD123 expression on K562 cells were (4.10 ± 1.46) ％,(7.2 ± 2.50) ％ as well as (21.6 ± 1.17) ％,respectively.Compared with the control group,107 U/L rhIFN-γ significantly increased the expression of CD123 on K562 cells (P＜0.05).Conclusion The effect of rhIFN-γ on the growth of K562 cells has two aspects (inhibition or proliferation),and it can increase the expression of CD 123.
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Objective:This study aimed to detect the expression of annexin A1 in human non-small cell lung cancer (NSCLC) and to explore its clinical significance. Methods:We detected the expression levels of annexin A1 in NSCLC and the same patient's dis-tal cancerous tissue (from the edge of the tumor >5 cm) by real-time fluorescence quantitative polymerase chain reaction (real-time PCR), Western blot, and immunohistochemical methods, and then analyzed their correlation with clinical pathological parameters. Re-sults: Real-time PCR showed that the expression of annexin A1 mRNA in NSCLC was higher than that of distal cancerous tissue (0.574±1.403 vs. 0.240±0.893, t=2.060, P=0.045). Moreover, the expression of annexin A1 in NSCLC was associated with the degree of differentiation, lymph node metastasis, and TNM staging (P0.05). Western blot and immunohistochemistry revealed that the expression of annexin A1 protein in NSCLC was higher than that of distal cancerous tissue. Conclusion:Annexin A1 is highly expressed in the cancer tissues of patients with NSCLC, which may be correlated with the occurrence and development of tumor and its invasion and metastasis.
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Objective To investigate the differentiation of mesenchymal stem cells(rMSCs) of young rat into neuron-like cells with Ligustrazin hydrochloride. Methods rMSCs were separated from femurs marrow flushed out with DMEM(low glucose) by using a needle and syringe, then planted in plastic culture flask. Through expanded to 5 passages, rMSCs were induced to differentiate into neuron-like cells with Ligustrazin hydrochloride. Anti-neurofilament(NF-M), nestin, neuron-specific enolase(NSE), MAP-2,GAP-43 and glial fibrillary acidic protein(GFAP) antibodies were detected by immunohistochemistry. Results rMSCs were comprised a single phenotypic population and displayed a fibroblast-like morphology after 5 passage in culture. With 10*!?g/L bFGF pre-induce for 24*!h, then the medium was replaced with induction medium containing Ligustrazin hydrochloride. The induced-rMSCs exhibited neuronal morphological characteristics from the first half an hour to 5*!h. The neuron-like cells expressed NF-M, NSE, MAP-2,GAP-43 and nestin positive, but didn't express glial astrocyte marker GFAP.Conclusion rMSCs can be induced to differentiate into neuron like cells with Ligustrazin hydrochloride in vitro.