Increased Co-Expression of Macrophage Migration Inhibitory Factor and Matrix Metalloproteinase 9 Is Associated with Tumor Recurrence of Meningioma

Background and Objective: We detected the expression of MIF and matrix metalloproteinase 9 (MMP9) in meningiomas to determine whether they are valuable recurrence predictor for meningioma.Methods: 67 cases of meningiomas, including 57 benign tumors (WHO grade I) and 10 non-benign tumors (WHO grade II and III), were collected, and expression of MIF and MMP9 in tissue microarray was evaluated immunohistochemically. The correlations between immunostainings and clinicopathological parameters, as well as the follow-up data of patients, were analyzed statistically.Results: Increased expressions of both MIF (58.2%, 39/67) and MMP9 (55.2%, 37/67) were significantly associated with microvessel density (MVD) of tumor, but only dual high-expression of MIF and MMP9 was in relation to tumor invasion (P=0.016) and tumor recurrence (P=0.001). Based on univariate analysis, histological grade, tumor invasion and co-expression of MIF and MMP9 were significant predictors for recurrence. However, only histological grade and co-expression of MIF and MMP9 in tumor were independent recurrence factors with a hazard ratio of 49.033 (P=0.002) and 37.766 (P=0.002) in multivariate analysis.Conclusions: Together with histological grade, increased co-expression of MIF and MMP9 in tumor might be a valuable predictor for recurrence, especially for benign meningiomas.     

The Incidence of Invasion and Metastasis of Nasopharyngeal Carcinoma at Different Anatomic Sites in the Skull Base

This study aimed to investigate the incidence of direct invasion and metastasis of nasopharyngeal carcinoma (NPC) at different anatomic sites in the skull base using magnetic resonance imaging (MRI). MRI data from 101 NPC patients with skull base invasion were collected and we analyzed the incidence and anatomic sites of invasion of NPC in the skull base. Of the 101 NPC patients, 84 had direct invasion at the skull base (83.2%), and 17 had skull base metastasis (16.8%). Affected sites with direct invasion in the skull base included sphenoid sinus and sella base, cavernous sinus, internal carotid canal, and clivus blumenbachii. Skull base metastasis sites included the internal carotid canal and jugular foramen area. Because of early lymphatic metastasis of NPC to the skull base, MRI examination can be helpful in increasing the accuracy of diagnostic imaging for skull base invasion of NPC and selecting appropriate target sites, radiotherapy techniques, and operative approaches. Anat Rec, 2012. © 2012 Wiley Periodicals, Inc.     

心绞痛患者血浆C反应蛋白及巨噬细胞移动抑制因子的变化

目的通过检测稳定型及不稳定型心绞痛患者血浆高敏c反应蛋白（hsCRP）及巨噬细胞移动抑制因子（MIF），了解hsCRP及MIF在其中的变化。方法61例稳定型心绞痛患者、69例不稳定型心绞痛患者和55例健康志愿者。以ELISA法检测分析其血浆中高敏C反应蛋白及巨噬细胞移动抑制因子的变化。结果不稳定型心绞痛组患者血浆hsCRP水平（12．93±3．67）mg／L及MIF水平（23．60±6．31）μg／L高于稳定型心绞痛患者[hsCRP（4．37±1．28）mg／L，MIF（9．89±1．04）μg／L]及对照组[hsCRP（3．51±1．47）mg／L，MIF（9．58±1．25）μg／L]，差异有统计学意义（P均〈0．05）；而稳定型心绞痛患者及对照组hsCRP及MIF水平比较差异则无统计学意义（P〉0．05）。结论不稳定型心绞痛患者其hsCRP及MIF水平较健康对照组及稳定型心绞痛患者均有明显升高，对hsCRP及MIF检测可能有助于鉴别高危冠心病患者。     

鼻咽癌组织中巨噬细胞移动抑制因子和MMP-2、MMP-9表达的关系

目的　研究巨噬细胞移动抑制因子 (macrophagemigrationinhibitoryfactor,MIF)和MMP 2、MMP 9在鼻咽癌组织中的表达水平及相互关系 ,探讨鼻咽癌细胞早期侵袭转移的机制。方法　收集 4 5例确诊的鼻咽原发癌活检组织标本 ,采用免疫组化LSAB法检测鼻咽癌组织中MIF和MMP 2、MMP 9的表达 ,并分析患者的临床参数的关系。结果　在 4 5例鼻咽原发癌组织中 ,MIF、MMP 2和MMP 9的阳性表达率分别为 77 8% (35 / 4 5 )、6 4 4 % (2 9/ 4 5 )和 71 1% (32 / 4 5 )。其中 ,癌细胞MIF和MMP 9的表达水平均显示与淋巴结转移有关 ,伴有淋巴结转移的癌组织中二者表达水平均高于无淋巴结转移的癌组织 (P值均 <0 0 5 )。MIF阳性组的癌细胞MMP 9的表达 (5 0 2 %± 33 5 % )明显高于MIF阴性病例 (11 7%± 2 2 7% ) ,两者差异有显著性 (P <0 0 1) ,且MIF的表达与MMP 9的表达亦呈正相关 (rs=0 .4 92 ,P <0 0 1) ,但癌细胞MMP 2的表达与MIF、MMP 9的表达以及是否有淋巴结转移则均未显示相关性。以Schmincke型生长方式分布的癌细胞MIF表达水平 (6 7 4 %±35 2 % )也高于以Regaud型方式分布的癌细胞 (32 9%± 2 9 7% ) ,差异有显著性 (P <0 0 1)。结论　鼻咽癌组织中癌细胞的MIF和MMP 9同步过表达 ,可能在鼻咽癌细胞的转移     

Epitope Mapping of Monoclonal Antibody 1B9 Against Plasmodium falciparum-Derived Macrophage Migration Inhibitory Factor

A homologue of mammalian macrophage migration inhibitory factor (MIF) has been identified in Plasmodium falciparum (PfMIF). This parasite-derived cytokine and its antiserum were detected in the circulation of patients with malaria. Using a monoclonal antibody, designated mAb1B9, against PfMIF, we performed biopanning using two phage display peptide libraries to screen for the main sequence of the epitope recognized by the antibody. We then expressed a series of truncated peptides in order to identify the precise sequence of the epitope. The epitope recognized by mAb1B9 is 22 amino acids long and has the following sequence: ³⁶LGYIMSNYDYQKNLRFGGSNEA⁵⁷. Western analysis showed that the residues ³⁶LG³⁷ and ⁵²G differentiated PfMIF from the rodent malaria parasite-derived MIFs, and the residues ⁴³Y and ⁴⁸NL⁴⁹ differentiated PfMIF from P. vivax- and P. knowlesi-derived MIFs. The precise identification of this epitope, the first identified for PfMIF, will increase the specificity of the sandwich ELISA assays used to evaluate patients with malaria. These results indicate that mAb1B9 is useful for investigating the function of PfMIF in immune responses to malaria. Both the epitope and the monoclonal antibody against it will be valuable tools in epidemiological studies concerning this P. falciparum-derived cytokine.     

Chemical Characterization of Macrophage Cytotoxicity Factor,Macrophage Migration Inhibitory Factor,T-Helper Cell-Replacing Factor and Colony-Stimulating Factor from Culture Supernatants of Concanavalin A-Stimulated Murine Spleen Cells

Supernatants from Concanavalin A-stimulated murine spleen cells were subjected to hydrophobic interaction chromatography on phenyl-Sepharose. Macrophage cytotoxicity factor (MCF), macrophage migration inhibitory factor (MIF), T-helper cell-replacing factor (TRF) and colony-stimulating factor (CSF) were bound at high ionic strength and were released stepwise at low ionic strength. CSF thus could be separated from MCF, MIF and TRF and the bulk of other proteins. Chromatography of pools containing MCF, MIF and TRF on Sephadex did not lead to a separation of the three activities which were all found in a molecular weight range of 25.000-55.000. Isoelectric focusing of these pools in pH range from 4 to 9 gave two peaks for MCF at pH 8.2 and 7.2, whereas MIF activity focused from pH 4.5 to 5.5. TRF activity was found in a single sharp peak at pH 5.3. The results demonstrate that the four biological activities can be distinguished on a chemical basis and are accessible for purification and chemical characterization.     

Reduced MTA1 Expression by RNAi Inhibits in Vitro Invasion and Migration of Esophageal Squamous Cell Carcinoma Cell Line

To distinguish aggressive esophageal squamous cell carcinoma from indolent disease is the important clinical challenge. Studies have indicated that metastasis-associated gene 1(Mta1) played a role in the process of metastasis of carcinoma. The overexpression of Mta1 gene has been found in a variety of tumors. To identify the detailed roles of MTA1 protein in the carcinogenesis of esophageal squamous cell carcinoma, this study analyzed the pathological specimens on tissue microarray derived from 72 patients using immunohistochemistry. MTA1 expression increased in the nuclear with the development of esophageal squamous cell carcinoma from normal epithelial cell, dysplasia, to invasive cancer. In biological studies with human esophageal squamous cell carcinoma cell line, MTA1 plays its roles to promote cancer cell invasion, adhesion and movement. RNA interference (RNAi) against MTA1 decreased the malignant phenotypes. Gene microarray analysis revealed some metastasis-associated genes were altered by MTA1 RNAi. This study started an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1.     

Increased expression of IRS-1 is associated with lymph node metastasis in nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is a head and neck malignant tumor rare throughout most of the world but common in Southeast Asia, especially in Southern China, which is with characteristics of early cervical lymph node metastasis and high incidence rate of distant metastasis. Insulin receptor substrate 1 (IRS-1) is a signaling adapter protein that is encoded by the IRS-1 gene in humans, plays an important role in the development, progression, invasion and metastasis of tumors. The aim of the present study was to investigate the association between the expression of IRS-1 protein and clinicopathological characteristics in NPC by immunohistochemistry. The results showed that the expression level of IRS-1 was significant higher in NPC than that in the control nasopharyngeal epithelia (P = 0.042). The positive percentage of IRS-1 expression in NPC with lymph node metastasis was also significantly higher than those without lymph node metastasis (P = 0.008). Positive expression of IRS-1 was proved to be the independent predicted factor for lymph node metastasis of NPC (P = 0.025) regardless of age, gender, histological type and clinical stages by multivariate logistic regression analysis. In addition, results showed higher sensitivity and agreement rate of IRS-1 for predicting lymph node metastasis of NPC patients. Taken together, high expression of IRS-1 might be closely correlated with lymph node metastasis in NPC and positive expression of IRS-1 could be used as an independent biomarker for predicting lymph node metastasis of NPC.     

Macrophage migration inhibitory factor deficiency protects pancreatic islets from cytokine-induced apoptosis in vitro

During pathogenesis of diabetes, pancreatic islets are exposed to high levels of cytokines and other inflammatory mediators that induce deterioration of insulin-producing beta cells. Macrophage migration inhibitory factor (MIF) plays a key role in the onset and development of several immunoinflammatory diseases and also controls apoptotic cell death. Because the occurrence of apoptosis plays a pathogenetic role in beta cell death during type 1 diabetes development and MIF is expressed in beta cells, we explored the influence of MIF deficiency on cytokine-induced apoptosis in pancreatic islets. The results indicated clearly that elevated MIF secretion preceded C57BL/6 pancreatic islets death induced by interferon (IFN)-γ + tumour necrosis factor (TNF)-α + interleukin (IL)-1β. Consequently, MIF-deficient [MIF-knock-out (KO)] pancreatic islets or islet cells showed significant resistance to cytokine-induced death than those isolated from C57BL/6 mice. Furthermore, upon exposure to cytokines pancreatic islets from MIF-KO mice maintained normal insulin expression and produced less cyclooxygenase-2 (COX-2) than those from wild-type C57BL6 mice. The final outcome of cytokine-induced islet apoptosis in islets from wild-type mice was the activation of mitochondrial membrane pore-forming protein Bcl-2-associated X protein and effector caspase 3. In contrast, these apoptotic mediators remained at normal levels in islets from MIF-KO mice suggesting that MIF absence prevented initiation of the mitochondrial apoptotic pathway. Additionally, the protection from apoptosis was also mediated by up-regulation of prosurvival kinase extracellular-regulated kinase 1/2 in MIF-KO islets. These data indicate that MIF is involved in the propagation of pancreatic islets apoptosis probably via nuclear factor-κB and mitochondria-related proteins.     

High-expression of DJ-1 and Loss of PTEN Associated with Tumor Metastasis and Correlated with Poor Prognosis of Gastric Carcinoma

Background and aims: DJ-1 and PTEN have been shown to involve in multiple cell processes and play an important role in cancer development and progression. However, their relationship with gastric carcinoma (GC) has not been identified yet. The purpose of this study is to clarify the relationship of DJ-1 and phosphatase and tensin homolog (PTEN) with clinicopathological parameters and prognosis in GC.Methods: 114 specimens were collected from GC patients and expression of DJ-1 and PTEN in tissue microarray was evaluated by immunohistochemical staining. Correlation between immunostainings and clinicopathological parameters, follow-up data of patients, was analyzed statistically.Results: High expression of DJ-1 was found in 66.7% (76/114) and associated with tumor depth (P=0.003), lymph node metastasis (P=0.011), distant metastasis (P=0.001) and advanced clinical stage (P=0.001). Loss or downregulation of PTEN was found in 58.7% (67/114) and associated with advanced clinical stage (P=0.018) and high expression of DJ-1 in tumor cells (P=0.006). In univariate survival analysis, high-expression of DJ-1 or loss of PTEN was significantly associated with poor prognosis of GC patients. However, only tumor depth (P=0.011) and coexistence of DJ-1 and PTEN abnormal expression (P=0.009) emerged as strong independent prognostic factors for overall survival of GC patients.Conclusions: the present study indicates that DJ-1 and PTEN may play their roles in progression of GC in a cooperating pattern. Co-existence of abnormal DJ-1 and PTEN expression is likely to serve as an independent predictive factor for prognosis of GC patients.     

VEGF和MMP-2的表达对肝细胞癌侵袭转移的影响

采用免疫组化 ABC法对 5 0例肝细胞癌手术切除标本的血管内皮生长因子 (VEGF)和基质金属蛋白酶- 2 (MMP- 2 )表达进行检测 ,以探讨 VEGF及 MMP- 2在肝细胞癌中的表达与其复发、转移的关系。结果发现 VEGF和 MMP- 2在肝细胞癌组织中的阳性表达率分别为 86 % (43/5 0 )和 6 0 % (30 /5 0 ) ,在正常肝组织中分别为 5 3.3%(16 /30 )和 30 % (9/30 ) ,癌组织与正常组织间存在着显著差异 (P<0 .0 5 ) ;VEGF和 MMP- 2的阳性表达率与肝细胞癌的转移和包膜形成相关 ,在肝细胞癌生长、侵袭和转移过程中起着重要的作用 ,对预测肝细胞癌复发、转移具有重要的意义     

巨噬细胞移动抑制因子在大肠黏膜癌变患者中表达增强

目的 探讨巨噬细胞移动抑制因子(MIF)表达与大肠癌临床病理因素的关系.方法 采用免疫组化法测定大肠组织中MIF的表达.ELISA测定血清中MIF水平.结果 MIF在正常大肠黏膜、大肠腺瘤、大肠癌中阳性表达强度和阳性表达率依次增高,3组之间存在显著性差异(P<0.001);组织中MIF表达强度与血清中MIF水平呈正相关性;MIF的表达与大肠癌的分化程度、淋巴结转移、肝转移有关.结论 MIF可能在大肠癌的发生和进展中起着重要的作用.     

Production of Five Human Lymphokines (GranulocyteMacrophage Colony Stimulating Factor,Interferon-γ, Interleukin 2, Macrophage Cytotoxicity Factor and Macrophage Migration Inhibitory Factor) from Con A Stimulated Lymphocyte Cultures in Bioreactors

Cultivation parameters for the production of five lymphokines, granulocyte-macrophage colony stimulating factor (GM-CSF), interferon-y (IFN-γ), interleukin 2 (IL-2), macrophage cytotoxicity factor (MCF), and macrophage migration inhibitory factor (MIF) from human spleen cells or peripheral blood lymphocytes were optimized. Cultivation was done in bioreactors containing up to 200 ml of medium, usually serum-free. The reactors were equipped with surface aeration facilities, stirrers and oxygen electrodes.Whereas stirring speed alone did not influence the yields of lymphokines, good aeration was especially beneficial for high IL-2 yields. However, all lymphokines were also produced under anaerobic conditions. The concentration of the mitogen concanavalin A was mainly critical for optimal IL-2 release. Optimal cell concentrations varied from 5 x 10₆/ml (for GM-GSF and MCF) to 10 x 10₆/ml (for IL-2 and IFN-γ). It was possible to increase the yields of individual lymphokines 3 to 10-fold per batch of lymphocytes by a reinduction procedure which involved a change of medium and mitogen every 24 hrs. Reinduction was possible up to 4 times, especially when serum was present in the culture media.     

糖尿病肾病合并冠心病患者C反应蛋白及巨噬细胞移动抑制因子的改变

目的通过检测糖尿病肾病合并冠心病患者血浆高敏C反应蛋白（hsCRP）及巨噬细胞移动抑制因子（MIF），了解hsCRP及MIF在糖尿病肾病合并冠心病患者中的变化，协助临床决策。方法收入64例糖尿病肾病合并冠心病患者及57例糖尿病肾病但无明确冠心病依据的患者，同时收入54例健康志愿者，以ELISA法检测分析其血浆中高敏C反应蛋白及巨噬细胞移动抑制因子的变化。结果糖尿病肾病合并冠心病组患者血浆hsCRP水平（9．93±2．58）mg／L及MIF水平（23．61±6．64）μg／L高于无明确冠心病依据的患者[hsCRP（7．37±1．32）mg／L，MIF（15．56±3．70）μg／L]，差异有统计学意义（P均〈0．05），两组患者血浆hsCRP及MIF水平又均显著高于对照组[hsCRP（3．51±2．00）mg／L，MIF（9．57±1．25）μg／L]。结论检测糖尿病肾病患者血浆hsCRP及MIF，可能有助于鉴别糖尿病肾病患者有无合并冠心病的危险。     

Upregulation of LMP1 Expression by Histone Deacetylase Inhibitors in an EBV Carrying NPC Cell Line

OBJECTIVES: In about 60% of Epstein-Barr virus (EBV) carrying nasopharyngeal carcinomas (NPC) LMP1 expressing cells can be detected. The frequency of LMP1 positive cells and the expression level varies from cell to cell in the different tumors. Cell lines derived from EBV positive NPCs loose the virus during in vitro culture. The in vitro infected NPC cell line TWO3-EBV used in our study carries the neomycin-resistance gene containing EBV and expresses low level of LMP1. With this cell line it was thus possible to study the regulation of LMP1 expression by modification of chromatin acetylation state. STUDY DESIGN: The TWO-EBV cell line was treated with n -butyrate (NB) or trichostatin A (TSA). RESULTS: Shown by immunoblotting, the LMP1 level was elevated in the treated samples. Already 2 h after TSA exposure LMP1 expression was higher and it increased up to 24 h. Immunofluorescence staining showed that nearly all cells were LMP1 positive. Neither EBNA2 nor BZLF1 were induced. Tested first 2 h after the treatment, acetylated histone H3 and H4 were already detectable, and their level increased up to 8 h. Chromatin immunoprecipitation (ChIP) verified that the LMP1-promoter (LMP1p) (ED-L1) was acetylated after TSA treatment. CONCLUSION: EBV carrying epithelial cells do not express EBNA-2. We showed that LMP1 expression was upregulated by histone deacetylase inhibitors in an in vitro infected, EBV carrier NPC cell line.     

Low EphA7 Expression Correlated with Lymph Node Metastasis and Poor Prognosis of Patients with Esophageal Squamous Cell Carcinoma

As a member of the Eph family of receptor tyrosine kinases, EphA7 plays an important role in cancer. However, the expression and significance of Eph receptors in esophageal squamous cell carcinoma (ESCC) remain unclear. Here, we detected the expression of EphA7 by immunohistochemistry in a sample of 352 patients with ESCC, and aimed to investigate the expression status of EphA7 in ESCC and its impact on prognosis. The results showed that low EphA7 expression significantly correlated with lymph node metastases (N0: 29%; N1: 64%. p<0.001), poor degree of tumor differentiation (G1: 31%; G2: 49%; G3: 58%. p=0.009) and pTNM staging (I+II: 33%; III+IV: 58%. p<0.001). Furthermore, in a combined analysis, patients with low EphA7-expressing tumors showed a shorter overall survival than those with high expression, resulting in a five-year overall survival rate of 47.4% vs. 52.6%, respectively (p=0.016). Consequently, patients with a low EphA7 expression have poorer prognosis in ESCC compared with those manifesting high expression.     

syndecan-1表达对肝细胞癌增殖、血管生成和转移的影响

目的探讨syndecan-1在肝细胞癌（hepatocellular carcinoma,HCC）表达的临床病理意义,及其与肿瘤转移、增殖和血管生成的关系。方法用免疫组织化学染色检测了syndecan-1在30例HCC原发癌组织、癌旁组织和肝内转移灶的表达,血管内皮生长因子（vascularendothelial growth factor,VEGF）和CD34-微血管密度（microvessel density,MVD）在原发灶和肝内转移灶的表达,及Ki-67在原发灶的表达。结果与HCC癌旁组织相比,syndecan-1在原发癌组织表达水平明显增高,syndecan-1在肝内转移灶表达水平明显下降（χ2=25.62,P〈0.001）。在HCC原发癌组织中,syndecan-1表达强度与Ki-67指数呈正相关（r=0.386,P〈0.05）。在肝内转移灶中,syndecan-1表达强度与CD34-MVD呈正相关（r=0.370,P〈0.05）。结论 HCC组织syndecan-1表达可能影响肿瘤转移、增殖和血管生成,提示syndecan-1在HCC生长和转移中的特殊作用可能为HCC有效治疗策略提供新思路。     

CD44v4在乳腺浸润性导管癌中的表达及 其与浸润转移的关系

目的 探讨CD44v4在浸润性乳腺导管癌中的表达及其与浸润转移的关系。方法 选取2011年1月~2018年3月我院收治的乳腺浸润性导管癌185例设为观察组,另选取同期收治的乳腺良性疾病患者60例设为对照组,采用免疫组化法检测两组CD44v4的表达情况,并比较观察组CD44v4表达及临床病理因素情况。结果 观察组CD44v4阳性表达率为60.54%,高于对照组的5.00%,差异有统计学意义（P＜0.05）。CD44v4表达在不同年龄、绝经前后月经状况、原发瘤大小间比较,差异无统计学意义（P＞0.05）;不同TNM分期、组织学分级、淋巴结转移患者的CD44v4表达比较,差异有统计学意义（P＜0.05）。结论 CD44v4蛋白在乳腺浸润性导管癌患者中为高表达,检测其表达可作为判断肿瘤预后的新指标。