Associations of content and gene polymorphism of macrophage inhibitory factor-1 and chronic hepatitis C virus infection

BACKGROUND The expression of macrophage inhibitory factor-1(MIC-1) is increased in peripheral blood of patients with chronic hepatitis and liver cirrhosis. However, whether MIC-1 gene polymorphism is correlated with relevant diseases is not yet reported.AIM To explore the correlation between gene polymorphism in MIC-1 exon region and chronic hepatitis C virus(HCV) infection.METHODS This case-control study enrolled 178 patients with chronic hepatitis C(CHC) in the case group, and 82 healthy subjects from the same region who had passed the screening examination comprised the control group. The genotypes of rs1059369 and rs1059519 loci in the MIC-1 gene exon were detected by DNA sequencing. Also, the MIC-1 level, liver function metrics, liver fibrosis metrics, and HCV RNA load were determined. Univariate analysis was used to compare the differences and correlations between the two groups with respect to these parameters. Multivariate logistic regression was used to analyze the independent relevant factors of CHC.RESULTS The plasma MIC-1 level in the CHC group was higher than that in the control group(P < 0.05), and it was significantly positively correlated with alanine aminotransferase, aspartate aminotransferase(AST), type III procollagen N-terminal peptide(known as PIIINP), type IV collagen, and HCV RNA(P < 0.05), whereas negatively correlated with total protein and albumin(P < 0.05). The genotype and allele frequency distribution at the rs1059519 locus differed between the two groups(P < 0.05). The allele frequency maintained significant difference after Bonferroni correction(Pc < 0.05). Logistic multiple regression showed that AST, PIIINP, MIC-1, and genotype GG at the rs1059519 locus were independent relevant factors of CHC(P < 0.05). Linkage disequilibrium(LD) was found between rs1059369 and rs1059519 loci, and significant difference was detected in the distribution of haplotype A-C between the CHC and control groups(P < 0.05). Meanwhile, we found the MIC-1 level trend to increase among rs1059519 genotypes(P = 0.006) and the level of MIC-1 in GG genotype to be significantly higher than CC genotype(P = 0.009, after Bonferroni correction).CONCLUSION Plasma MIC-1 level was increased in CHC patients and correlated with liver cell damage, liver fibrosis metrics, and viral load. The polymorphism at the MIC-1 gene rs1059519 locus was correlated with HCV infection, and associated with the plasma MIC-1 level. G allele and GG genotype may be an important susceptible factor for CHC.

Mudskipper interleukin-34 modulates the functions of monocytes/macrophages via the colony-stimulating factor-1 receptor 1

Interleukin-34(IL-34)is a novel cytokine that plays an important role in innate immunity and inflammatory processes by binding to the colonystimulating factor-1 receptor(CSF-1R).However,information on the function of IL-34 in fish remains limited.In the present study,we identified an IL-34 homolog from mudskippers(Boleophthalmus pectinirostris).In silico analysis showed that the mudskipper IL-34(BpIL-34)was similar to other known IL-34 variants in sequence and structure and was most closely related to an orange-spotted grouper(Epinephelus coioides)homolog.BpIL-34 transcripts were constitutively expressed in various tissues,with the highest level of expression found in the brain.Edwardsiella tarda infection significantly up-regulated the mRNA expression of BpIL-34 in the mudskipper tissues.The recombinant mature BpIL-34 peptide(rBpIL-34)was purified and used to produce anti-rBpIL-34 IgG.Western blot analysis combined with PNGase F digestion revealed that native BpIL-34 in monocytes/macrophages(MOs/MФs)was N-glycosylated.In vitro,rBpIL-34 treatment enhanced the phagocytotic and bactericidal activity of mudskipper MOs/MФs,as well as the mRNA expression of pro-inflammatory cytokines like tumor necrosis factorα(BpTNF-α)and BpIL-1βin these cells.Furthermore,the knockdown of mudskipper CSF-1R1(BpCSF-1R1),but not mudskipper BpCSF-1R2,significantly inhibited the rBpIL-34-mediated enhanced effect on MO/MФfunction.In conclusion,our results indicate that mudskipper BpIL-34 modulates the functions of MOs/MФs via BpCSF-1R1.

Local inhibition of matrix metalloproteinases reduced M2 macrophage activity and impeded recovery in spinal cord transected rats after treatment with fibroblast growth factor-1 and nerve grafts

Alternatively activated macrophages （M2 macrophages） promote central nervous system regeneration. Our previous study demonstrated that treatment with peripheral nerve grafts and fibroblast growth factor-1 recruited more M2 macrophages and improved partial functional recovery in spinal cord transected rats. The migration of macrophages is matrix metalloproteinase （MMP） dependent. We used a general inhibitor of MMPs to influence macrophage migration, and we examined the migration of macrophage populations and changes in spinal function. Rat spinal cords were completely transected at Ts, and 5 mm of spinal cord was removed （group T）. In group R, spinal cord-transected rats received treatment with fibroblast grow th factor- 1 and peripheral nerve grafts. In group RG, rats received the same treatment as group R with the addition of 200 μM GM6001 （an MMP inhibitor） to the fibrin mix. We found that MMP-9, but not MMP- 2, was upregulated in the graft area of rats in group R. Local application of the MMP inhibitor resulted in a reduction in the ratio of arginase-1 （M2 macrophage subset）/inducible nitric oxide synthase-postive cells. When the MMP inhibitor was applied at 8 weeks postoperation, the partial functional recovery observed in group R was lost. This effect was accompanied by a decrease in brain-derived neurotrophic factor levels in the nerve graft. These results suggested that the arginase-1 positive population in spinal cord transected rats is a migratory cell population rather than the phenotypic conversion of early iNOS^＋ cells and that the migration of the arginase-1^＋ population could be regulated locally. Simultaneous application of MMP in- hibitors or promotion of MMP activity for spinal cord injury needs to be considered if the coadministered treatment involves M2 recruitment.

Serum and ascites levels of macrophage migration inhibitory factor, TNF-α and IL-6 in patients with chronic virus hepatitis B and hepatitis cirrhosis

Objective: To study the potential role of macrophage migration inhibitory factor (MIF), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the development of chronic virus hepatitis B (CH) and hepatitis cirrhosis (HC). Methods: The serum concentrations of MIF, TNF-α and IL-6 in 18 patients with chronic virus hepatitis B and in 14 patients with hepatitis cirrhosis without as- citic fluid, and the serum and ascites cytokine con- centrations in 22 HC patients with ascitic fluid were detected by enzyme linked immunity sorbed assay. Results: The cytokine concentrations of the patients were significantly higher than those of the controls. The serum levels of MIF, TNF-α and IL-6 of the 22 patients with ascitic fluid were higer than those of 14 HC patients without ascites. In the 18 patients with CH, the serum cytokine concentrations were the low- est. The serum cytokine concentrations of the 22 HC patients with ascites were significantly higher than those of the 14 HC patients without ascites (P< 0. 01). Their serum cytokine concentrations were sig- nificantly higher than those in the 18 patients with CH (P<0. 01). The concentration of IL-6 in ascites was the highest among all the groups. The serum le- vels of MIF, TNF-α and IL-6 are correlated with al- anine aminotransferase (ALT) in the patients with CH, but not in those with HC with or without asci- tes. Conclusions: These results indicated that MIF, TNF- α and IL-6 may participate in the pathological process of CH and cirrhosis, that IL-6 seems to play an important role in ascites formation, and that se- rum levels of MIF, TNF-α and IL-6 appear to reflect the severity of tissue injury in HBV disease.

Macrophage migration inhibitory factor regulates proliferation of gastric cancer cells via the PI3K/Akt pathway

AIM:To investigate the effects of macrophage migration inhibitory factor (MIF) on proliferation of human gastric cancer MGC-803 cells and expression of cyclin D1 and p27Kip1 in them,and further determine whether the effects are related to the PI3K/Akt signal transduction pathway. METHODS:Gastric cancer MGC-803 cells were cultured and then treated with 50 μg/L recombinant human MIF (rhMIF) with and without a PI3K inhibitor,LY294002 (25 μmol/L). MTT assay was used to detect the prolifer-ation of MGC-803 cells. Cell cycle was detected by flow cytometry. Expression of cyclin D1 and p27Kip1 mRNA was by reverse transcription-polymerase chain reaction. Protein expression of phosphorylated Akt (p-Akt),Akt,cyclin D1 and p27Kip1 was examined by immunocyto-chemistry and Western blotting. RESULTS:rhMIF signifi cantly stimulated the prolifera-tion of MGC-803 cells and cell cycle progression from G1 phase to S phase in a concentration-and time-de-pendent manner. After the MGC-803 cells were treated with rhMIF for 24 h,the expression of cyclin D1 was signifi cantly up-regulated compared with the cells not treated with rhMIF at both mRNA and protein levels(0.97 ± 0.02 vs 0.74 ± 0.01,P = 0.002; 0.98 ± 0.05 vs 0.69 ± 0.04,P = 0.003). The p27Kip1 was down-regulated but only statistically significant at the protein level. rhMIF significantly increased the expression of p-Akt,which reached the peak at 30 min,but did not affect the expression of Akt. However,LY294002 inhibited all the effects of rhMIF.CONCLUSION:Macrophage MIF increases the proliferation of gastric cancer cells,induces the expression of cyclin D1 at the transcriptional level and inhibits the expression of p27Kip1 at the post-transcriptional level via the PI3K/Akt pathway.

Role of Cyclin D1b in Inducing Macrophages Toward a Tumor-associated Macrophage-like Phenotype in Murine Breast Cancer

Objective:Tumor-associated macrophages(TAMs)of the M2 phenotype are frequently associated with cancer progression.Invasive cancer cells undergoing epithelial-mesenchymal transition(EMT)have a selective advantage as TAM activators.Cyclin D1b is a highly oncogenic splice variant of cyclin D1.We previously reported that cyclin D1b enhances the invasiveness of breast cancer cells by inducing EMT.However,the role of cyclin D1b in inducing macrophage differentiation toward tumor-associated macrophage-like cells remains unknown.This study aimed to explore the relationship between breast cancer cells overexpressing cyclin Dlb and TAMs.Methods:Mouse breast cancer 4T1 cells were transfected with cyclin D1b variant and co-cultured with macrophage cells in a Transwell coculture system.The expression of characteristic cytokines in differentiated macrophages was detected using qRT-PCR,ELISA and zymography assay.Tumor-associated macrophage distribution in a transplanted tumor was detected by immunofluorescence staining.The proliferation and migration ability of breast cancer cells was detected using the cell counting kit-8(CCK-8)assay,wound healing assay,Transwell invasion assay,and lung metastasis assay.Expression levels of mRNAs were detected by qRT-PCR.Protein expression levels were detected by Western blotting.The integrated analyses of The Cancer Genome Atlas(TCGA)datasets and bioinformatics methods were adopted to discover gene expression,gene coexpression,and overall survival in patients with breast cancer.Results:After co-culture with breast cancer cells overexpressing cyclin D1b,RAW264.7 macrophages were differentiated into an M2 phenotype.Moreover,differentiated M2-like macrophages promoted the proliferation and migration of breast cancer cells in turn.Notably,these macrophages facilitated the migration of breast cancer cells in vivo.Further investigations indicated that differentiated M2-like macrophages induced EMT of breast cancer cells accompanied with upregulation of TGF-β1 and integrinβ3 expression.Conclusion:Breast cancer cells transfected with cyclin D1b can induce the differentiation of macrophages into a tumor-associated macrophage-like phenotype,which promotes tumor metastasis in vitro and in vivo.

抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用

目的探讨敲低长链非编码RNA(lncRNA)牛磺酸上调基因1(TUG1)抑制核苷酸结合寡聚结构域样受体蛋白1(NLRP1)炎症小体在缓解阿尔茨海默病进展中的作用。方法选取9~10周龄遗传背景为C57/BL6的野生型小鼠(WT组,10只)或淀粉样前体蛋白(APP)/早老素1(PS1)转基因小鼠(30只)。APP/PS1转基因小鼠随机分为模型(model)组,模型+敲低lncRNA TUG1组[model+lncRNA TUG1短发夹RNA(shRNA)组]和model+shRNA非靶标(NT)组,每组10只。分别采集12周龄第1天(3月龄)和32周龄第1天(8月龄)小鼠外周血和脑皮质组织,并分离皮质中的原代小胶质细胞和原代星形胶质细胞,每个时间点每组5只小鼠。Real-time PCR分别测定3月龄和8月龄上述4个分组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和巨噬细胞移动抑制因子(MIF)mRNA的水平,以及原代星形胶质细胞中补体蛋白C1r和C1s mRNA的水平。ELISA法测定其外周血浆中MIF含量。对3月龄和8月龄小鼠脑皮质原代小胶质细胞和原代星形胶质细胞共培养。CCK-8法测定上述2种细胞的增殖能力。Western blotting分别测定3月龄和8月龄上述4个分组小鼠脑皮质组织中MIF、白细胞介素1β前体(pro-IL-1β)、凋亡相关斑点样蛋白(ASC)、Caspase-1(p20)、Caspase-1(full)、NLRP1及NLRP3蛋白的表达水平。采用免疫荧光染色法测定8月龄各分组小鼠脑皮质组织中β淀粉样蛋白(Aβ)表达。结果3月龄和8月龄时,与WT组小鼠相比,model组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF相对表达水平显著上调,原代小胶质细胞和原代星形胶质细胞增殖能力增强(P<0.05)。与model组相比,model+lncRNA TUG1 shRNA组小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF的相对表达水平显著降低,原代小胶质细胞和原代星形胶质细胞增殖能力降低(P<0.05)。与WT组相比,model组小鼠外周血浆中MIF含量显著升高;小鼠脑皮质组织中pro-IL-1β、ASC、Caspase-1(p20)、Caspase-1(full)、NLRP1以及NLRP3的蛋白表达水平显著升高;Aβ免疫荧光强度明显增强(P<0.05)。与model组相比,model+lncRNA TUG1 shRNA组小鼠外周血浆中MIF含量显著降低;小鼠脑皮质组织中pro-IL-1β、ASC、Caspase-1(p20)、Caspase-1(full)和NLRP1的蛋白表达水平显著降低,Aβ免疫荧光强度明显降低(P<0.05),而NLRP3蛋白质的表达水平无明显变化(P>0.05)。与model组相比,model+shRNA NT组小鼠上述所有检测指标差异均无显著性(P>0.05)。结论APP/PS1转基因小鼠脑皮质组织和原代小胶质细胞中lncRNA TUG1和MIF因子表达上调与脑皮质内NLRP1炎症小体激活成正相关,敲低lncRNA TUG1可缓解阿尔茨海默病的进展。

Salidroside Inhibits Lipopolysaccharide-ethanol-induced Activation of Proinflammatory Macrophages via Notch Signaling Pathway

Activation of macrophages is a key event for the pathogenesis of various inflammatory diseases.Notch signaling pathway recently has been found to be a critical pathway in the activation of proinflammatory macrophages.Salidroside (Sal),one of main bioactive components in Rhodiola crenulata (Hook.F.et Thoms) H.ohba,reportedly possesses anti-inflammatory activity and ameliorates inflammation in alcohol-induced hepatic injury.However,whether Sal regulates the activation of proinflammatory macrophages through Notch signaling pathway remains unknown.The present study investigated the effects of Sal on macrophage activation and its possible mechanisms by using both alcohol and lipopolysaccharide (LPS) to mimic the microenvironment of alcoholic liver.Detection of THP-1-derived macrophages exhibited that Sal could significantly decrease the expression of tumor necrosis factor-α(TNF-α),interleukinbeta (IL-1β)and IL-6 in the macrophages at both mRNA and protein levels.Furthermore,Sal significantly suppressed NF-kB activation via Notch-Hes signaling pathway in a dose-dependent manner.Moreover,in the microenvironment of alcoholic liver,the expression of Notch-dependent pyruvate dehydrogenase phosphatase 1 (PDP1) was elevated,and that of Ml gene expression [inducible NO synthase (NOS2)] was up-regulated.These changes could all be effectively ameliorated by Sal.The aforementioned findings demonstrated that Sal could inhibit LPS-ethanol-induced activation of proinflammatory macrophages via Notch signaling pathway.

Macrophage secretory products induce an inflammatory phenotype in hepatocytes

AIM:To investigate the influence of macrophages on hepatocyte phenotype and function.METHODS:Macrophages were differentiated from THP-1 monocytes via phorbol myristate acetate stimulation and the effects of monocyte or macrophageconditioned medium on HepG2 mRNA and protein expression determined.The in vivo relevance of these findings was confirmed using liver biopsies from 147 patients with hepatitis C virus(HCV)infection.RESULTS:Conditioned media from macrophages,but not monocytes,induced a transient morphological change in hepatocytes associated with upregulation of vimentin(7.8±2.5-fold,P=0.045)and transforming growth factor(TGF)-β1(2.6±0.2-fold,P<0.001)and downregulation of epithelial cadherin(1.7±0.02-fold,P=0.017)mRNA expression.Microarray analysis revealed significant upregulation of lipocalin-2(17-fold,P <0.001)and pathways associated with inflammation,and substantial downregulation of pathways related to hepatocyte function.In patients with chronic HCV,realtime polymerase chain reaction and immunohistochemistry confirmed an increase in lipocalin-2 mRNA(F0 1.0 ±0.3,F1 2.2±0.2,F2 3.0±9.3,F3/4 4.0±0.8,P= 0.003)and protein expression(F1 1.0±0.5,F2 1.3± 0.4,F3/4 3.6±0.4,P=0.014)with increasing liver injury.High performance liquid chromatography-tandem mass spectrometry analysis identified elevated levels of matrix metalloproteinase(MMP)-9 in macrophageconditioned medium,and a chemical inhibitor of MMP-9 attenuated the change in morphology and mRNA expression of TGF-β1(2.9±0.2 vs 1.04±0.1,P<0.001) in macrophage-conditioned media treated HepG2 cells.In patients with chronic HCV infection,hepatic mRNA expression of CD163(F0 1.0±0.2,F1/2 2.8±0.3,F3/4 5.3±1.0,P=0.001)and MMP-9(F0 1.0±0.4,F1/2 2.8±0.3,F3/4 4.1±0.8,P=0.011)was significantly associated with increasing stage of fibrosis.CONCLUSION:Secreted macrophage products alter the phenotype and function of hepatocytes,with increased expression of inflammatory mediators,suggesting that hepatocytes actively participate in liver injury.

血清MIF、MCP-1、suPAR水平与脓毒症严重程度及合并ARDS风险的关系

目的探讨血清巨噬细胞迁移抑制因子(MIF)、单核细胞趋化蛋白-1(MCP-1)、可溶性尿激酶型纤溶酶原激活物受体(suPAR)水平与脓毒症严重程度及合并急性呼吸窘迫综合征(ARDS)风险的关系。方法回顾性分析2022年2月至2023年5月太原钢铁(集团)有限公司总医院收治的86例脓毒症患者的临床资料。依据病情程度不同将患者分为脓毒症组(n=20)、严重脓毒症组(n=48)和脓毒症休克组(n=18)。入院72 h内参考ARDS诊断标准将患者分为ARDS组(n=27)和非ARDS组(n=59)。检测并比较各组脓毒症患者血清MIF、MCP-1、suPAR水平。收集ARDS组与非ARDS组患者年龄、性别、体重指数、合并症、感染类型、既往史、心率、急性生理学和慢性健康状况评价Ⅱ(APACHEⅡ)、脓毒症相关性器官衰竭评价(SOFA)评分、白细胞计数、血乳酸、天冬氨酸转移酶(AST)、丙氨酸转移酶(ALT)、总胆固醇等指标。采用多因素Logistic回归分析对影响脓毒症患者并发ARDS的危险因素进行分析。通过受试者工作特征(ROC)曲线分析血清MIF、MCP-1、suPAR水平预测脓毒症患者并发ARDS的价值。结果脓毒症休克组患者血清MIF、MCP-1、suPAR水平分别为(94.02±10.13)、(506.55±45.15)、(13.89±3.95)ng/mL,均高于脓毒症组[(76.93±7.01)、(148.38±35.74)、(6.07±2.13)ng/mL]和严重脓毒症组[(85.46±8.74)、(327.08±40.62)、(8.42±1.07)ng/mL],而严重脓毒症组患者血清MIF、MCP-1、suPAR水平均高于脓毒症组,差异均有统计学意义(P<0.05)。ARDS组与非ARDS组患者的年龄、性别构成比、体重指数、合并症、感染类型、白细胞计数、心率、吸烟史、饮酒史、血乳酸、AST、ALT、总胆固醇比较,差异均无统计学意义(P>0.05);ARDS组患者APACHEⅡ评分、SOFA评分、有急腹症和胰腺炎占比及血清MIF、MCP-1、suPAR水平均高于非ARDS组,差异均有统计学意义(P<0.05)。经多因素Logistic回归分析结果显示,急腹症、胰腺炎、APACHEⅡ评分、SOFA评分、MIF、MCP-1、suPAR是影响脓毒症患者并发ARDS的独立危险因素(P<0.05)。经ROC曲线分析结果显示,血清MIF、MCP-1、suPAR水平均能预测脓毒症患者ARDS的发生,曲线下面积分别为0.904、0.910、0.917,预测价值较好(P<0.05)。结论血清MIF、MCP-1、suPAR水平与脓毒症患者病情程度、并发ARDS密切相关,且血清MIF、MCP-1、suPAR水平对ARDS的发生有较好的预测价值。

超声内镜、巨噬细胞抑制因子-1、糖类抗原19-9联合诊断胰腺癌的临床价值分析

目的探究超声内镜、巨噬细胞抑制因子-1(MIC-1)、血清肿瘤标志物糖类抗原19-9(CA19-9)联合诊断胰腺癌的临床价值。方法选取2019年3月至2022年3月在南京医科大学附属苏州市立医院行超声内镜检查的100例高度疑似胰腺癌患者作为观察组,同期来院进行体检的50例健康志愿者作为对照组。采用化学发光法检测CA19-9水平,采用酶联免疫法检测MIC-1水平并进行组间比较。以术后患者的病理检查结果为金标准,比较超声内镜、MIC-1、CA19-9分别及联合应用对胰腺癌的诊断价值。结果2组研究对象的血清CA19-9、MIC-1水平比较差异有统计学意义(P<0.05)。与健康对照组相比较,观察组患者的血清CA19-9、MIC-1水平更高,差异有统计学意义(P<0.05)。超声内镜、MIC-1、CA19-9三者联合诊断的敏感度(84.21%)均低于三者分别诊断(92.11%、90.79%、90.79%),特异度(91.67%)均高于三者分别诊断(79.17%、83.33%、75.00%)。三者联合诊断的阳性预测值为96.97%,阴性预测值为64.71%。结论在对胰腺肿瘤进行良恶性诊断时,采用超声内镜、MIC-1、CA19-9联合诊断可以有效提高其诊断效能。

高迁移率族蛋白B1巨噬细胞转移抑制因子联合CXC亚家族趋化因子16对高危型HPV感染患者诊断价值

目的:探究高迁移率族蛋白B1(HMGB1)、巨噬细胞转移抑制因子(MIF)联合CXC亚家族趋化因子16(CXCL16)在高危型人乳头状瘤病毒(HPV)感染患者中的表达意义及诊断价值。方法:纳入本院2020年10月至2023年10月期间接诊的150例HPV感染患者作为研究组,根据患者核酸检测结果分为高危型HPV感染组(n=40)和低危型HPV感染(n=110)。另选取同期于本院接受经阴道镜活检检查的健康女性作为对照组(n=100)。对研究组患者均采用HPV-DNA分型试剂盒检测;对研究组和对照组人员采用酶联免疫吸附法检测HMGB1和CXCL16水平,免疫组化检测MIF染色强度。对比对照组和研究组HMGB1、MIF和CXCL16差异;对比低危型和高危型HPV感染组HMGB1、MIF和CXCL16差异;采用受试者特征曲线(ROC)分析HMGB1、MIF和CXCL16联合检测对高危型HPV感染诊断价值;Spearman相关性分析HMGB1、MIF和CXCL16与不同HPV感染分型相关性。结果:与对照组相比,研究组血清HMGB1、CXCL16水平和MIF阳性细胞占比得分显著更高(均P<0.05);与低危型HPV感染组相比,高危型HPV感染组血清HMGB1、CXCL16水平和MIF阳性细胞占比得分显著更高(均P<0.05);ROC工作曲线分析结果显示HMGB1、MIF和CXCL16单独及联合诊断高危型HPV感染的AUC分别为0.748、0.684、0.791和0.934,联合检测诊断价值显著高于单独检测(Z=2.577、3.152、2.096,均P<0.05);Spearman相关性结果显示HMGB1、MIF和CXCL16与不同HPV感染分型存在显著正相关(r=0.615、0.633、0.649均P<0.05)。结论:高危型HPV感染患者血清HMGB1、CXCL16水平和MIF阳性细胞占比得分显著高于低危型和对照组,临床检查中应用HMGB1、MIF和CXCL16联合检测可提升高危型HPV感染临床诊断率,为临床提供一种新型检测方法和治疗依据。

lncRNA MIF-AS1调节miR-423-5p/PYCR1轴对前列腺癌细胞恶性生物学行为的影响

目的探究长链非编码RNA(lncRNA)巨噬细胞移动抑制因子反义RNA1(MIF-AS1)调节miR-423-5p/吡咯啉-5-羧酸还原酶1(PYCR1)轴对前列腺癌(PC)细胞恶性生物学行为的影响。方法体外培养PC3细胞,敲低MIF-AS1或下调miR-423-5p表达,检测PC患者肿瘤组织与癌旁组织和细胞中MIF-AS1、miR-423-5p和PYCR1 mRNA的表达;检测细胞增殖、凋亡、迁移与侵袭;Western blot检测PYCR1蛋白表达;验证miR-423-5p和MIF-AS1、PYCR1的关系。结果MIF-AS1、PYCR1 mRNA在肿瘤组织中呈高表达,miR-423-5p呈低表达。沉默MIF-AS1可抑制PC3细胞增殖、迁移、侵袭及PYCR1表达,上调miR-423-5p表达,诱导细胞凋亡(P<0.05);抑制miR-423-5p表达可逆转沉默MIF-AS1对PC3细胞恶性行为的抑制作用(P<0.05)。MIF-AS1、PYCR1与miR-423-5p存在靶向调控关系。结论沉默MIF-AS1可能通过上调miR-423-5p来抑制PYCR1表达,抑制PC细胞的恶性行为。

巨噬细胞抑制因子-1与早期非小细胞肺癌诊断及预后相关性

背景与目的巨噬细胞抑制因子-1(macrophage inhibitory cytokine-1,MIC-1)是人转化生长因子β(transforming growth factor-β,TGF-β)超家族中重要成员,研究发现MIC-1表达水平在多种上皮来源肿瘤患者血清中均有显著升高。本研究旨在探讨MIC-1在早期非小细胞肺癌(non-small cell lung cancer,NSCLC)诊断及其与临床病理特征间的关系,以及与术后复发/转移及预后的相关性。方法采用酶联免疫吸附试验(enzymelinked immunosorbent assay,ELISA)方法检测152例早期肺癌、48例肺良性疾病患者及105例正常对照人群血清MIC-1浓度,分析MIC-1诊断肺癌中的作用,同时分析血清MIC-1浓度与临床病理特征、复发/转移及预后的相关性。结果早期肺癌患者组MIC-1血清水平高于正常对照组(P<0.001)和肺良性疾病组(P<0.001),设1,000 pg/m L为诊断肺癌的临界值,MIC-1检测肺癌的敏感性和特异性分别为70.4%和99.0%[曲线下面积(area under curve,AUC):0.90;95%CI:0.87-0.94];MIC-1血清水平与年龄(P=0.001)、性别(P=0.03)有关,病理TNM分期T2的患者MIC-1血清水平高于T1患者(P=0.022);血清MIC-1>1,465 pg/m L组的患者3年生存率为77.6%,低于血清MIC-1<1,465 pg/m L组的患者94.8%(P=0.022),Cox回归多因素分析结果显示,血清MIC-1>1,465 pg/m L是I期、II期NSCLC独立的预后因素(HR=3.37,95%CI:1.09-10.42,P=0.035)。结论 MIC-1作为血清肿瘤生物标志物,可能有助于提高肺癌早期诊断。MIC-1的检测对判断I期、II期NSCLC患者预后有预测价值,可能为其独立的预后指标。

ISO-1对大肠癌细胞侵袭的影响

目的研究选择性MIF互变异构酶活性抑制剂ISO-1对人大肠癌细胞侵袭的影响并探讨其可能的机制。方法实验组用0．01-100μmol／L的ISO-1作用于大肠癌细胞Lovo，对照组为相应浓度的DMSO处理；MIF互变异构酶活性通过L-多巴色素甲酯测定；微孔迁移法检测ISO-1对Lovo细胞体外侵袭的影响；ELISA法测定ISO-1作用后培养上清中MIF蛋白水平；逆转录聚合酶链反应（RT-PCR）检测ISO-1对Lovo细胞MMP-9和IL-8 mRNA表达的影响。结果较高浓度ISO-1（10-100μmol／L）均能明显抑制MIF互变异构酶活性，但不影响细胞外分泌MIF蛋白水平；100μmol／L ISO-1作用24h Lovo细胞穿透聚碳酸酯膜显著减少，与对照组比较差异有显著性（153±13vs204±10，P〈0．05）；100μmol／L ISO-1作用24h Lovo细胞表达MMP-9和IL-8 mRNA水平显著降低，与对照组比较差异有显著性（MMP-9 mRNA：0．5187±0．072vs0．6757±0．026．P〈0．05；IL-8 mRNA：0．1541±0．019vs0．2081±0．030，P〈0．05）。结论选择性MIF活性抑制剂ISO-1降低了Lovo细胞的侵袭，其可能机制为ISO-1抑制了MIF互变异构酶活性并下调MMP-9和IL-8的表达。

MIC-1、VEGF和P53蛋白表达与直肠癌临床病理及预后的关系

背景与目的:巨噬细胞是重要的免疫效应细胞,对肿瘤的发生、发展发挥着重要的调控作用。本研究探讨巨噬细胞抑制细胞因子-1(MIC-1)、血管内皮生长因子(VEGF)、P53蛋白的表达与直肠癌临床病理特征及预后的相关性。方法:用免疫组化法检测73例直肠癌中MIC-1、VEGF和P53的表达,并与临床病理因素及预后进行相关性分析。结果:MIC-1表达与VEGF、P53表达呈正相关(P<0.05);MIC-1、VEGF和P53表达与直肠癌肿瘤临床分期、淋巴结转移呈明显相关性(P<0.01);MIC-1和VEGF、P53阳性组和阴性组的5年生存率差异均有显著性(P<0.01)。结论:MIC-1、VEGF和P53与直肠癌淋巴结转移、分期及预后有密切关系,淋巴结转移是最重要的独立的预后因素。

IGF-1、HMGB-1、GSN和MIF水平在高血压脑出血中的意义

目的观察血清胰岛素样生长因子(IGF)-1、高迁移率族蛋白(HMGB)-1、凝溶胶蛋白(GSN)和巨噬细胞移动抑制因子(MIF)检测在高血压性脑出血患者中意义。方法选择2013年1月至2015年6月诊治的高血压性脑出血患者78例,为高血压性脑出血组。根据神经功能损伤程度分为轻度17例,中度36例和重度25例。根据脑出血量分为:小量出血组36例,中量出血组27例和大量出血组15例。根据格拉斯哥预后评分将出院患者分为:预后良好组43例和预后不良组35例。选择同期就诊单纯脑出血患者和单纯高血压患者分别为脑出血组(45例)和高血压组(35例)。选择同期在我院健康体检者30例为健康对照组。观察高血压性脑出血组,脑出血组,高血压组和健康对照组的血清IGF-1、HMGB-1、GSN和MIF水平,高血压性脑出血患者血清IGF-1、HMGB-1、GSN和MIF水平与神经功能缺损评分和脑出血面积的关系,各指标与预后和各指标之间的相关性分析。结果高血压脑出血组的IGF-1和GSN水平明显低于脑出血组,高血压组和健康对照组(P<0.01),高血压组和脑出血组的水平明显低于健康对照组(P<0.01);高血压脑出血组的HMGB-1和MIF水平明显高于脑出血组,高血压组和健康对照组(P<0.01),高血压组和脑出血组的水平明显高于健康对照组(P<0.01)。高血压脑出血患者的IGF-1和GSN水平随着神经损害程度和脑出血面积的增加而降低(P<0.01),预后良好组的IGF-1和GSN水平明显高于预后不良组(P<0.01),而HMGB-1和MIF水平神经损害程度和脑出血面积增加而升高(P<0.01),预后良好组的HMGB-1和MIF水平明显低于预后不良组(P<0.01)。并发现IGF-1水平与HMGB-1(r=-0.457,P<0.05)和MIF(r=-0.536,P<0.05)呈负相关,与GSN呈正相关(r=0.754,P<0.05);HMGB-1水平与GSN呈负相关(r=-0.486,P<0.05),与MIF呈正相关(r=0.864,P<0.05),GSN与MIF呈负相关(r=-0.758,P<0.05)。结论 IGF-1、HMGB-1、GSN和MIF参与了高血压性脑出血疾病的发生发展,对于判断高血压脑出血的病情和预后具有重要意义。

不对称二甲基精氨酸对THP-1源性巨噬细胞表达巨噬细胞移动抑制因子的影响

目的研究不对称二甲基精氨酸对THP-1源性巨噬细胞表达巨噬细胞移动抑制因子的影响。方法 150nmol/L佛波酯诱导THP-1细胞48h使其转化为巨噬细胞,并用抗人CD68进行免疫细胞化学鉴定,以不同浓度的不对称二甲基精氨酸(1、5、10、20和30μmol/L)作用于THP-1源性巨噬细胞24h及20μmol/L不对称二甲基精氨酸作用于THP-1源性巨噬细胞0h、6h、12h、24h及48h,逆转录聚合酶链反应检测各组细胞的巨噬细胞移动抑制因子mRNA表达水平,酶联免疫吸附法检测各组细胞上清液中巨噬细胞移动抑制因子的蛋白含量。结果与对照组相比,5、10、20和30μmol/L的不对称二甲基精氨酸作用于THP-1源性巨噬细胞24h后,巨噬细胞移动抑制因子的mRNA和蛋白表达水平均明显提高(P<0.05)。与0h组相比,20μmol/L不对称二甲基精氨酸作用于THP-1源性巨噬细胞12、24和48h,巨噬细胞移动抑制因子的mRNA和蛋白表达水平均明显提高(P<0.05)。结论不对称二甲基精氨酸可上调THP-1源性巨噬细胞的巨噬细胞移动抑制因子表达,并呈剂量和时间依赖性,不对称二甲基精氨酸可能通过诱导巨噬细胞移动抑制因子的表达而促进动脉粥样硬化的发生和发展。

巨噬细胞抑制因子-1及其联合多种标志物在肺癌早期诊断和诊断中的临床意义

目的研究巨噬细胞抑制因子-1(macrophage inhibitory cytokine-1,MIC-1)在早期肺癌诊断中的辅助价值,并评价多种肿瘤标志物联合应用的临床意义。方法应用MIC-1定量检测试剂盒及Roche Cobas 601电化学发光免疫分析仪分别检测663例未经治疗的不同临床分期的肺癌患者和488例正常人群血清样本中的MIC-1、CEA、CA125、NSE、SCC和CYFRA21-1水平和分布,分析患者血清MIC-1水平与肺癌临床分期、病理分型和细胞分化程度的关系,并研究多种标志物联合检测的价值。结果肺癌患者血清MIC-1水平显著高于正常人群(P<0.001);MIC-1水平随临床分期的进展呈上升的趋势(P<0.001),且与肿瘤浸润(P<0.001)、淋巴结转移(P=0.02)、远端转移(P<0.001)和肿瘤分化程度(P<0.001)显著相关。单一检测MIC-1诊断肺癌的敏感度(76.6%)高于其他五种肺癌标志物的联合应用(72.2%),且MIC-1在鳞癌、腺癌、小细胞癌诊断中的敏感度均能达到甚至超过其他五种标志物的联合诊断水平(81.6%vs 82.8%;74.7%vs 68.9%;84.9%vs 83.0%)。在肺癌早期,以MIC-1为主的六种肿瘤标志物联合检测的诊断敏感度(Ⅰ期:79.8%;Ⅱ期:87.7%)显著高于其他五种肿瘤标志物联合诊断的敏感度(Ⅰ期:44.9%;Ⅱ期:72.6%)。结论 MIC-1是肺癌,尤其是早期肺癌有价值的血清肿瘤标志物,MIC-1和CEA、CA125、NSE、SCC、CYFRA21-1联合检测用于普通人群体检和肺癌早期诊断具有重要的临床意义和价值。

巨噬细胞抑制因子-1联合多种标志物在结直肠癌诊断中的价值研究

目的研究巨噬细胞抑制因子1(MIC-1)在结直肠癌诊断和早期诊断中的价值,并探讨多种标志物联合应用于检测结直肠癌的可行性。方法应用自主研制的MIC-1定量检测试剂盒及Roche Cobas 601电化学发光免疫分析仪分别检测429例不同临床分期的结直肠癌患者和129例健康人血清样本中的MIC-1、CA19-9、CEA和TPS水平,比较MIC-1水平与其他肿瘤标志物的关系、与患者TNM分期的关系,以及在结直肠癌诊断和早期诊断中的价值。结果结直肠癌患者血清中的MIC-1水平显著高于正常对照人群(1045.88±892.67 vs 398.04±263.19,P<0.001);肿瘤患者血清MIC-1水平随肿瘤浸润深度(T分期)增加而升高(P=0.001),并与淋巴结转移(N分期,P=0.007)和远端转移显著正相关(P<0.001);MIC-1、CEA和TPS三种标志物联合检测早期结直肠癌检测敏感性可达61.3%,显著高于CEA的检出率(25.6%)。结论 MIC-1是结直肠癌尤其是早期结直肠癌有价值的血清肿瘤标志物,MIC-1、CEA和TPS联合检测对于提高结直肠癌的早期诊断率具有重要的临床意义和价值。