Expression of midkine in the cochlea

Midkine (MK) is one of a new family of heparin-binding growth factors involved in the regulation of growth and differentiation. We have analyzed expression of MK in the cochlea using ICR mice within 1 day from birth. The expression of MK in the cochlea was confirmed by Western blotting and immunohistochemistry. Anti-MK immunoreactivity was observed in the stria vascularis, spiral prominence, spiral ganglion, and ganglion nerve fibers. These findings suggest that MK plays a role in the development of the cochlea.     

喉癌病人外周血中期因子水平与喉癌临床特征及预后的关系

目的　研究喉癌病人外周血中期因子 (midkine,MK)水平与喉癌临床特征及预后的关系。方法　采用酶联免疫吸附测定检测喉癌病人及健康体检者 (对照组 )外周血中MK水平 ,通过SPSS软件分析MK与喉癌临床特征及预后的关系。结果　喉癌病人血清MK水平为 (40 0± 2 2 9) pg/ml,对照组为(1 1 8± 97) pg/ml,差异有统计学意义 (P <0 .0 0 1 )。以 30 0 pg/ml为分界值 ,MK阳性率随喉癌进展呈上升趋势 (P <0 .0 1 ) :I期 5 3%(1 0 / 1 9) ,Ⅱ期 6 0 %(1 8/ 30 ) ,Ⅲ期 6 2 %(2 8/ 4 5 ) ,Ⅳ期 75 %(1 5 / 2 0 )。MK水平与T分期成正相关 (P <0 .0 1 )。随访病例 5 8例 ,经Kaplan Meier生存曲线分析 ,5年生存率MK阳性者为 5 3%,阴性者为 77%,差异有统计学意义(P <0 .0 5 )。结论　高水平的MK(≥ 30 0 pg/ml)与不良预后相关。血清MK水平可以作为预测喉癌病人的生存期的一个临床指标。     

MK因子在不同病理类型宫颈癌组织的表达及临床意义

目的:探讨MK因子(中期因子)在宫颈癌的表达与临床病理特征的关系。方法:用免疫组织化学法(SP法)检测178例宫颈癌组织和74例宫颈上皮内瘤变组织中MK因子的表达。结果:178例宫颈癌组织中,MK蛋白阳性表达率为91.57%,明显高于宫颈上皮内瘤样病变组织的14.86%(P<0.001)。宫颈癌临床分期不同MK蛋白阳性表达率不同,临床Ⅰ期阳性表达率显著低于临床Ⅱ~Ⅳ期(P<0.001)。宫颈鳞癌MK阳性表达率显著高于腺癌(P<0.001),但与组织分化程度无关(P=0.123)。有盆腔淋巴结转移者MK阳性表达率显著高于无盆腔淋巴结转移者(P=0.026)。结论:MK因子阳性表达与宫颈癌病理特征关系密切,可能成为一种有价值的肿瘤标志物,对宫颈癌的早期诊断及治疗有一定的临床指导意义。     

MK和CD105在大肠癌中的表达及其临床意义

 目的探讨MK和CD105的表达与大肠癌生物学行为的关系及对预后的意义。方法采用免疫组织化学方法检测50例大肠癌组织MK和CD105表达水平,并对其与大肠癌临床病理特征的关系进行统计学分析。结果本组50例中,MK阳性表达率为72%。MK为阳性时CD105标记的MVD值(71.73±6.99)明显高于MK为阴性组的MVD值(66.09±7.49)(P<0.05)。两者的表达均与大肠癌的Duke's分期、淋巴结转移、浸润深度有关。结论MK的阳性表达与大肠癌的发生、发展和预后密切相关,可联合CD105作为一组有价值的肿瘤标记和预后指标。     

Midkine, a new neurotrophic factor, is present in glial cytoplasmic inclusions of multiple system atrophy brains

The glial cytoplasmic inclusion (GCI) is a histological hallmark for multiple system atrophy (MSA): these inclusions are found in oligodendrocytes and consist of abnormal granule-coated fibrils of approximately 24- to 40-nm diameter. To clarify the significance of the presence of midkine (MK) in these GCIs, we carried out immunohistochemical, electron and immunoelectron microscopical, and Western blot analyses of MSA brains using a monoclonal antibody against the C-terminal region of human MK. Immunohistochemically, most of the GCIs were intensely stained by the antibody to MK. Electron and immunoelectron microscopy showed that the GCIs were composed of MK-positive granule-coated fibrils that were essential constituents of these inclusions. No significant MK immunoreactivity was observed in oligodendrocytes, astrocytes and neurons of the normal control subjects. The presence of MK in MSA brain but not in normal brain was confirmed by Western blotting. Together with the fact that MK is associated with fetal morphogenesis during the midgestation period, the presence of MK immunoreactivity in oligodendroglial GCIs may suggest the existence of a repair mechanism on the basis of morphogenesis in the degenerated oligodendrocytes themselves as well as the affected neurons and their axons through the oligodendrocyte-axon-neuron relationship. Received: 8 November 1999 / Revised, accepted: 31 January 2000     

中期因子与脑膜瘤血管生成关系的研究

目的:探讨中期因子(MK)蛋白与脑膜瘤中血管生成关系。方法:采用S-P免疫组织化学技术,检测52例脑膜瘤组织和10例正常脑膜组织中MK蛋白的表达和微血管密度(MVD),并对二者的关系进行统计学分析。结果:52例脑膜瘤组织中,MK蛋白阳性表达率为63.5%,正常脑膜组织中无MK蛋白表达,明显低于肿瘤组织,差异有显著性(P<0.05)。MK蛋白阳性表达程度与MVD呈正相关(r=0.756,P<0.05),与肿瘤的血供(r=0.6518,P<0.05)密切相关。结论:MK蛋白在脑膜瘤组织中有过度表达;MK蛋白的表达与脑膜瘤血管生成相关;MK蛋白有望成为临床上对脑膜瘤生物学行为进行判定的指标。抑制MK的表达,有望为高表达MK的脑膜瘤提供一个新的治疗途径。     

Development of bead based multiplexed immunoassay for evaluation of midkine,syndecan-1, and ANGPTL4 in patient serum

Background: Angiogenesis is associated with tumor progression in a range of malignancies. Herein, we develop custom immunobead assays for several mechanistically important targets and evaluated these against sera from cohorts of non-small cell lung cancer (NSCLC) patients.

Methods: Antigen “capture” antibodies for midkine, syndecan-1, and ANGPTL4 were independently conjugated to MagPlex® Microspheres using standard carbodiimide/NHS-based chemistry. These reagents served as the basis for quantitative sandwich assay assembly using biotinylated detection antibodies and R-phycoerythrin-conjugated streptavidin reporter system. Standard curves were created using dilution series of recombinant target proteins with assay performance characteristics calculated, accordingly. Finally, we evaluated a range of serum samples from NSCLC patients (n = 32) to verify assay performance.

Results: Multiplexed assays for midkine, syndecan-1, and ANGPTL4 were developed with three orders of magnitude in dynamic range, excellent intra- and inter-assay precision, and accuracy parameters (<10%, and <15% variability, respectively). Detection and quantifications limits were suitable for the three assays to efficiently evaluate sera across a range of disease stages with a four-fold dilution factor.

Conclusion: We successfully developed and analytically validated a 3-plex immunobead assay for quantifying midkine, syndecan-1, and ANGPTL4 in patient sera. This multiplexed assay will provide an important tool for future studies delineating the role of angiogenesis in lung cancer progression.     

Identification and validation of a poor clinical outcome subtype of primary prostate cancer with Midkine abundance

Recent studies identified Midkine (MDK) as playing a key role in immune regulation. In this study, we aimed to discover the clinical significance and translational relevance in prostate cancer (PCa). We retrospectively analyzed 759 PCa patients who underwent radical prostatectomy from Huashan Hospital, Fudan University (training cohort, n = 369) and Chinese Prostate Cancer Consortium (validation cohort, n = 390). A total of 325 PCa patients from The Cancer Genome Atlas (TCGA) database (external cohort) were analyzed for exploration. Immune landscape and antitumor immunity were assessed through immunohistochemistry and flow cytometry. Patient‐derived explant culture system was applied for evaluating the targeting potential of MDK. We found that intratumoral MDK expression correlated with PCa progression, which indicated an unfavorable biochemical recurrence (BCR)‐free survival for postoperative PCa patients. Addition of MDK expression to the postoperative risk assessment tool CAPRA‐S could improve its prognostic value. Tumors with MDK abundance characterized the tumor‐infiltrating CD8⁺ T cells with less cytotoxicity production and increased immune checkpoint expression, which were accompanied by enriched immunosuppressive contexture. Moreover, MDK inhibition could reactivate CD8⁺ T cell antitumor immunity. MDK mRNA expression negatively correlated with androgen receptor activity signature and positively associated with radiotherapy‐related signature. In conclusion, intratumoral MDK expression could serve as an independent prognosticator for BCR in postoperative PCa patients. MDK expression impaired the antitumor function of CD8⁺ T cells through orchestrating an immunoevasive microenvironment, which could be reversed by MDK inhibition. Moreover, tumors with MDK enrichment possessed potential sensitivity to postoperative radiotherapy while resistance to adjuvant hormonal therapy of PCa. MDK could be considered as a potential therapeutic target for PCa.     

人胃腺癌BGC823细胞Midkine高表达前后基因表达谱芯片分析

目的：用基因芯片技术分析、筛选Midkine高表达前后胃腺癌相关基因谱的差异表达。方法：建立Midkine高表达BGC823胃腺癌细胞系,提取细胞总RNA,逆转录Cy3、Cy5标记cDNA探针,应用晶芯唧表达谱芯片,分析差异表达的基因。结果：发现Midkine高表达前后BGC823胃腺癌细胞有显著性表达差异的基因共有550个,其中上调基因407个,下调基因143个。对550条差异表达基因进行初步功能分类分析,结果表明这些基因与Midkine促进胃腺癌发病机制存在相关性。结论：Midkine高表达前后BGC823胃腺癌细胞部分功能基因存在显著的表达差异,多数差异表达的基因参与细胞黏附、细胞骨架形成、细胞周期以及细胞代谢等过程。     

中期因子对心肌梗死大鼠心肌胶原代谢的影响

目的探讨中期因子（Midkine,MK）对大鼠心肌胶原代谢的影响。方法 40只Wistar大鼠随机分为4组,每组10只：空白组（Control组）、伪手术组（Sham组）、梗死模型组（MI组）、MK治疗组（MI＋MK组）。通过结扎大鼠左冠状动脉前降支制作大鼠心肌梗死模型,MI＋MK组于心梗后立即给予MK心肌注射（1μg/只,分五个点注射）。4周后,各组剩余大鼠采血检测血清基质金属蛋白酶-9（MMP-9）、Ⅲ型前胶原氨基端肽（PⅢNP）水平;取心脏称重,计算全心肥厚指数;将心肌行冠状切片,行Masson染色,测定各组大鼠梗死区厚度、长度、梗死面积、梗死区及非梗死区胶原容积分数（CVF）。结果 MI组较Control、Sham组血清MMP-9、PⅢNP水平明显增高（P〈0.05）;MI＋MK组较MI组血清中MMP-9水平明显降低（P〈0.05）,PⅢNP水平明显增高（P〈0.05）;MI＋MK组较MI组梗死区厚度明显增厚,长度明显变短,梗死面积明显变小（P〈0.05）;MI＋MK组较MI组梗死区胶原容积分数明显增高（P〈0.05）,非梗死区胶原容积分数明显减低（P〈0.05）。结论 MK能够有效减轻心肌纤维化程度,其机制是通过调节心肌梗死区及非梗死区的胶原代谢来实现的。