肝细胞癌患者外周血中Midkine、CTCs的表达水平及临床意义

目的:观察中期因子（Midkine）、循环肿瘤细胞（CTCs）在肝细胞癌患者外周血中的表达水平,并研究它们与肝癌临床病理参数之间的关系。方法:收集我院96例原发性肝癌患者临床资料为肝癌组,40例健康体检组作为对照组。采用酶联免疫吸附法（ELISA）和电化学发光法检测Midkine水平,采用阴性富集法检测CTCs计数。分析Midkine、CTCs在肝细胞癌患者外周血中的表达与临床病理参数之间的关系以及肝细胞癌患者外周血Midkine表达与CTCs表达的相关性。结果:肝癌组外周血Midkine的表达水平显著高于对照组（P＜0.001）;肝癌组外周血中CTCs阳性检出率(70.8%,68/96)显著高于对照组(7.5%,3/40)（P＜0.001）。肝癌患者外周血Midkine表达在肿瘤大小、血管侵犯、Child-pugh分级、AFP水平、BCLC分期及Edmondson-Steiner分级均有显著差异（P＜0.05）。肝癌患者外周血CTCs表达在血管侵犯、Child-pugh分级、AFP水平、BCLC分期及Edmondson-Steiner分级均有显著差异（P＜0.05）。本研究结果显示,96例肝癌患者中,上皮型CTCs阳性率为31.3%,间质型CTCs阳性率为44.8%,混合型CTCs阳性率为67.7%。三种CTCs类型在肿瘤大小、血管侵犯、AFP水平、BCLC分期及Edmondson-Steiner分级均有显著差异（P＜0.05）。Spearman相关性分析得出,外周血Midkine表达与CTCs表达呈正相关（r=0.531,P=0.007）。结论:肝细胞癌患者外周血Midkine与CTCs的表达均高于正常组,Midkine与CTCs可能在肝癌的发生发展中发挥作用,且两者在肝细胞癌患者外周血中的表达呈正相关。     

妊娠中肾细胞因子mRNA在人食管癌组织中的表达

目的 研究妊娠中肾细胞因子（midkine, MK)mRNA在食管癌组织中的表达。方法 用Trizol提取食管癌组织和相应癌旁正常组织RNA，经RT－PCR获得扩增的MK cDNA，溴化乙锭琼脂糖凝胶电泳检测PCR产物。结果 6例食管癌组织在450bp处均出现一条MK的特征条带，其中1例低分化食管癌MK条带最深。6例癌旁正常组织均检测不出MK mRNA。此外，2例食管癌组织除显示MK的特征条带外，在280bp处出现一条截短型MK（tMK)区带。结论 MK在食管癌组织特异表达，表达程度可能与细胞分化程度相关。     

Midkine expression in colorectal tumors: correlation with Ki-67 labeling in sporadic, but not ulcerative colitis-associated ones

Midkine (MK) is a heparin-binding growth factor encoded by a retinoic acid responsive gene. To investigate the possible contribution of MK to genesis of colorectal carcinomas, an immunohistochemical examination of protein expression was conducted in sporadic and ulcerative colitis (UC)-associated tumors. MK expression significantly differed among normal mucosa, adenomas with low-grade dysplasia (LGD), adenomas with high-grade dysplasia (HGD) and invasive adenocarcinomas: MK expression was increased along with tumor progression. UC-associated lesions (regenerative mucosa of UC, UC-associated dysplasia and UC-associated adenocarcinoma) had similar variations. MK expression in UC-associated lesions was significantly higher than in normal mucosa, although there was no significant difference among UC-associated lesions. However, in UC-associated dysplasia, MK expression did not differ between the upper and lower halves, in contrast to adenoma with LGD and HGD, in which MK expression was significantly higher in the upper than lower halves, corresponding to cell proliferative zone. Furthermore, correlations with Ki-67 and single-strand DNA labeling, respectively, reflecting cellular proliferative activity and apoptosis, were noted in sporadic but not UC-associated lesions. These results suggest that MK is involved in genesis/development of sporadic colorectal tumors as well as of UC-associated tumors, but might contribute differently to genesis/development in these two types of tumors.     

MK对人脐静脉内皮细胞增殖及迁移的影响

目的 通过研究midkine（MK）蛋白对人脐静脉内皮细胞（HUVEC）增殖、迁移的影响,阐明MK蛋白在血管生成中的作用.方法 选择HUVEC为研究对象进行细胞常规培养,分为5组分别给予不同浓度MK进行干预,实验组MK浓度分别为0.5、5、50和500 ng/ml,以未处理组作为对照组.利用CCK-8试剂盒检测MK对HUVEC增殖能力的影响,利用Transwell技术检测MK对HUVEC迁移能力的影响.结果 与对照组相比,5 ng/ml的MK作用24h和48 h可以促进细胞增殖,差异有统计学意义（P＜0.05）;5 ng/ml的MK作用24 h可以促进细胞迁移,差异有统计学意义（P＜0.05）.结论 一定浓度的MK作用于内皮细胞,可以促进其增殖和迁移,提示MK在血管生成中具有重要作用.     

中期因子在喉癌中的表达及其临床意义

目的 检测中期因子(Midkine,MK)在喉癌组织中的表达,探讨MK高表达与喉癌颈淋巴结转移及预后的关系。方法 应用免疫组化法检测喉癌组织中MK的表达水平及微血管密度(MVD)。结果 14例喉正常黏膜和14例乳头状瘤组织中未见MK蛋白染色或仅见微弱染色(-)(2例乳头状瘤),表达率为0;而在75例喉癌中35例弱阳性(+),21例强阳性(++),表达率为74.7%。肿瘤细胞的MK与生存时间有关(P=0.001),MK表达从阴性(-)到阳性(+、++)生存期较原来下降5.63倍,应用多因素分析结果表明淋巴结的有无与复发有关,有淋巴结转移的复发是无淋巴结转移的5.488倍。结论 MK在喉癌组织中的高表达与颈淋巴结转移密切相关,可以作为喉癌诊断、预后的判定指标。     

中期因子和CD105在下咽癌组织中的表达及其临床意义

目的探讨中期因子MK和CD105在下咽鳞状细胞癌组织中的表达及临床意义。方法采用免疫组织化学方法检测47例下咽鳞状细胞癌组织中MK的表达和CD105标记的微血管密度MVD情况,并根据检测结果分析与下咽癌各临床病理因素之间的关系。结果 MK的阳性表达率为68.08%,MK表达为阳性时CD105标记的MVD值(37.21±7.94)明显高于MK表达为阴性时的MVD值(25.11±7.94)(P<0.01),二者表达与临床分期、淋巴结转移有关。结论 MK及CD105标记的微血管密度(MVD)与下咽癌分期及淋巴结转移具有相关性,与下咽癌的分化程度无相关性,可作为一组有价值的肿瘤标记和预后指标。     

中期因子在宫颈癌组织中表达的临床意义

目的 探讨中期因子(MK )在宫颈癌组织中表达的临床意义.方法 采用免疫组化链霉菌抗生物素蛋白一过氧化酶联结法(SP)检测134例宫颈癌、42例宫颈上皮内瘤样病变和46例慢性宫颈炎组织MK的阳性表达情况,比较不同宫颈疾病组织中MK表达有无差异及比较宫颈癌组织中MK表达在不同临床分期、组织分化程度、有无淋巴转移病理类型有无差异. 结果 慢性宫颈炎、宫颈上皮内瘤样病变和宫颈癌组织中MK的阳性表达率分别为8.7%、26.8%和76.1%,宫颈癌组织中MK的阳性表达率高于慢性宫颈炎、宫颈上皮内瘤样病变,差异有统计学意义(P＜0.05),慢性宫颈炎组织与宫颈上皮瘤样病变组织中MK阳性表达率比较,差异有统计学意义(P＜0.05).宫颈癌组织中MK的阳性表达率与宫颈癌组织分化程度、临床分期和淋巴结转移有关,差异均有统计学意义(P＜0.05).结论 MK可能与宫颈癌的发生、发展有关,对宫颈癌的早期诊断及治疗有一定的临床指导意义.     

Midkine and its receptor in regenerating rat skeletal muscle after bupivacaine injection

Midkine (MK) is a multifunctional cytokine and heparin-binding growth factor with neurotrophic activity. MK and its receptor were examined for up to 14 days in a chemically injured rat muscle regeneration process caused by the injection of bupivacaine using immunohistochemical and Western blot analysis. Although MK immunoreactivity was not detectable in the mature uninjured skeletal muscle, MK was strongly detected in the regenerating muscle cells. MK immunoreactivity was observed in the myoblast-like cells and myotubes, which were desmin-positive cells, whereas it was not detectable in the surviving normal muscle fibers. Most myotubes labeling for desmin showed MK immunoreactivity 5-7days after the injury. However, MK immunoreactivity was not detected 14 days after the injury. Immunoreactivity of low-density lipoprotein receptor-related protein (LRP), a cell membrane receptor of MK, was detected in the regenerating muscle cells, whereas it was not detected in the normal adult skeletal muscle and surviving muscle. These findings suggested that MK was involved. MK may have a role for differentiation during skeletal muscle regeneration and may be taken up in an autocrine fashion with LRP.     

Increased serum midkine levels in autism spectrum disorder patients

Background: Midkine (MK) is a heparin binding growth factor and is involved in neurogenesis, neural development and neuroprotection. Additionally, MK may contribute to cancer development and pathogenesis of neurodegenerative disorders and schizophrenia. Considering these effects of MK, this study researched whether MK is involved in autism spectrum disorders (ASD) pathogenesis.

Methods: We evaluated serum MK levels of 38 patients with ASD and 32 healthy control group. MK levels were measured with ELISA, while ASD severity was assessed with Childhood Autism Rating Scale.

Results: Our data showed that the serum MK concentration in ASD patients (mean ± SD, 11.51 ± 8.53 pg/ml) is significantly higher than healthy controls (mean ± SD, 6.19 ± 3.94 pg/ml) (p = 0.007).

Conclusions: According to these results, MK may play a role in ASD pathogenesis.     

The growth factor midkine may play a pathophysiological role in rheumatoid arthritis

Objectives: Midkine (MK) is involved in cell proliferation, differentiation, migration, and survival. In this study, we measured serum MK levels in rheumatoid arthritis (RA) and investigated the correlation of serum MK with RA disease activity. Expression and effect of MK in RA synovial tissue were also examined.

Methods: Serum MK and production of inflammatory mediators by rheumatoid synovial fibroblasts (RSFs) were measured by enzyme-linked immunosorbent assay. MK expression in synovial tissue was examined by immunohistochemistry. MK receptor expression was analyzed by RT-PCR and Western blotting.

Results: RA patients had a significantly higher serum MK level than healthy controls. In RA patients, the MK level was correlated with DAS28-ESR, disability index of the Health Assessment Questionnaire, and rheumatoid factor level. The serum MK level tended to be decreased by anti-TNF therapy. MK was expressed by synovial lining cells in RA synovial tissues and it enhanced the production of IL-6, IL-8, and CCL2 by RSFs. RSFs expressed LDL receptor-related protein 1, candidate receptor for MK.

Conclusions: The serum MK level could be a marker of disease activity in RA and an indicator of a poor prognosis. MK may have a role in the pathogenesis of RA via induction of inflammatory mediators.