血清与腹腔液中趋化因子RANTES水平在子宫内膜异位症患者中的临床应用

目的:探讨血清与腹腔液中趋化因子RANTES水平在子宫内膜异位症(EM)患者中的临床意义。方法:选取2012年5月-2013年5月本院收治的33例EM患者(观察组)、33例良性卵巢肿瘤患者(对照组)和33例健康体检者(健康对照组),应用ELISA法对血清与腹腔液中趋化因子RANTES水平进行检测,分析RANTES水平与患者r-AFS分期及痛经程度的相关性。结果:观察组血清RANTES水平明显高于对照组和健康对照组,差异均有统计学意义(t=7.163,6.743,均P0.05);观察组腹腔液RANTES水平亦高于对照组,两组比较差异有统计学意义(t=5.927,P0.05);观察组血清及腹腔液中RANTES水平与r-AFS分期呈正相关(r=0.975,0.893,均P0.05),且随分期增高而呈递增趋势;观察组血清RANTES水平与患者痛经评分无明显的相关性(r=-0.312,P0.05);而腹腔液中RANTES水平与患者痛经评分呈正关(r=0.517,P0.05)。结论:EM患者血清与腹腔液中趋化因子RANTES水平明显上升,应用ELISA法检测RANTES水平可辅助EM诊断,有利于提高诊断准确率。     

子宫内膜异位症患者血清可溶性B7-H4的水平及意义

目的:探讨子宫内膜异位症患者血清可溶性B7-H4(sB7-H4)的水平及其临床意义。方法:用ELISA夹心法检测43例子宫内膜异位症患者术前血清sB7-H4的水平及40例子宫内膜异位症患者术后血清sB7-H4的水平,同时选取30例体检健康妇女血清sB7-H4水平作为对照。结果:子宫内膜异位症患者血清sB7-H4水平为(36.23±5.67)μg/L,体检健康者血清sB7-H4水平为(31.24±4.56)μg/L,两者比较差异有统计学意义(P0.01)。手术前,子宫内膜异位症患者血清sB7-H4水平为(36.23±5.67)μg/L,明显高于术后(32.54±4.27)μg/L(P0.05)。子宫内膜异位症患者血清sB7-H4水平与CA125水平呈显著正相关(r=0.531,P0.01)。结论:血清可溶性B7-H4可能与子宫内膜异位症的发病有关,检测血清中可溶性B7-H4水平对内异症的辅助诊断和疗效观察可能具有一定的临床意义。     

趋化因子受体CCR3在上皮性卵巢癌中的表达及临床意义

目的:探讨趋化因子受体CCR3在上皮性卵巢癌组织中的表达情况,以及其与卵巢癌临床病理特征的关系。方法:收集上皮性卵巢癌组织、良性上皮性卵巢肿瘤组织以及正常卵巢组织标本各30例,采用多聚腺苷酸加尾实时荧光定量反转录聚合酶链反应[poly(A)-RT-qPCR]检测其CCR3的表达,并分析上皮性卵巢癌组织中CCR3的表达与患者临床病理特征之间的关系。结果:上皮性卵巢癌组织中CCR3的表达显著高于良性卵巢肿瘤组和正常卵巢组,差异有统计学意义(P0.05);且上皮性卵巢癌组织中CCR3的表达与患者的分期、组织分级及淋巴结转移均显著相关(P0.05)。结论:CCR3在上皮性卵巢癌组织中呈高表达,且在上皮性卵巢癌的发生和发展过程中均起着十分重要的作用。     

膀胱癌组织中CXCR4 和CXCR7 的表达及临床意义

目的:探讨膀胱癌组织中趋化因子受体4(CXCR4)和趋化因子受体7(CXCR7)的表达及临床意义。方法:收集2012年1月至2014年1月我院收集的膀胱癌组织标本96例,肿瘤旁正常组织标本42例,采用免疫组化方法检测组织标本中CXCR4和CXCR7的表达情况。结果:96例癌组织中检出CXCR4阳性59例,阳性率为61.46%,检出CXCR7阳性表达71例,阳性率为73.96%;42例癌旁组织中检出CXCR4阳性11例,阳性率26.19%,检出CXCR7阳性8例,阳性率为19.05%,癌组织与癌旁组织中CXCR4和CXCR7的表达具有统计学差异(均P0.05);相关性分析显示在膀胱癌组织中,CXCR4和CXCR7的表达呈正相关性(r=0.497,P=0.001);CXCR4和CXCR7在浸润性高(T2-T3)的膀胱癌和分化程度低(G2-G3)的膀胱癌表达强度较高,且差异具有统计学意义(均P0.05)。结论:CXCR4和CXCR7协同参与了膀胱癌的发生发展,并且与肿瘤的分化程度和浸润程度密切相关,有望成为诊断和治疗的重要靶点,在临床应用上具有重要意义。     

ER和PRmRNAs在内异症子宫内膜表达的变化

目的 :探讨雌激素受体 (ER)和孕激素受体 (PR)在子宫内膜异位症 (内异症 )子宫内膜的表达。方法 :利用大鼠内异症动物模型 ,采用逆转录聚合酶链反应 (RT PCR)技术 ,检测子宫内膜ER和PRmRNAs的表达情况。结果 :内异症模型组大鼠异位内膜ER、PRmRNAs的表达低于在位内膜和对照组正常子宫内膜 ,与后两者比较差异有显著性意义 (P <0 .0 1) ;而模型组在位内膜ER、PRmRNAs的表达与正常对照组比较差异无显著性意义 (P >0 .0 5 )。内异症模型组异位内膜ER/PRmRNA比值大于在位内膜和正常子宫内膜ER/PRmRNA比值 (P <0 .0 1)。结论 :内异症大鼠异位内膜ERmRNA表达的相对增高在内异症的发生与发展中起着一定的作用。     

P物质在子宫内膜异位症中的表达及其意义

目的:探讨P物质(substance P,SP)在子宫内膜异位症(endometriosis,EM)中的表达及其意义。方法:采用免疫组织化学法检测2012年10月至2013年4月在哈尔滨医科大学附属第一医院经腹腔镜及病理证实的EM患者异位子宫内膜组织20例,与其配对的在位子宫内膜组织10例以及因非EM(子宫肌瘤)行腹腔镜下子宫全切术或肌瘤核除患者的正常子宫内膜组织20例中SP的表达情况,并分析和比较术中盆腔粘连发生情况。结果:SP在EM患者的异位子宫内膜、在位子宫内膜及非EM患者的正常子宫内膜的阳性表达率分别为75%、80%、20%,异位子宫内膜和在位子宫内膜比较无显著性差别(P=1.0),但均高于非EM患者的正常子宫内膜,差异均有统计学意义(P=0.002,P=0.004);EM组盆腔粘连的阳性率高于非EM组,EM患者异位子宫内膜中SP阳性组盆腔粘连阳性率高于SP阴性组,差异均有统计学意义(P=0.001,P=0.032),非EM患者正常子宫内膜中SP阳性组盆腔粘连阳性率与SP阴性组相比,差异不具有统计学意义(P=0.061)。结论:SP在EM的异位子宫内膜和在位子宫内膜中的表达上调,并与EM合并盆腔粘连有关,其具体机制尚有待进一步的研究。     

子宫内膜异位症患者在位内膜及异位内膜中Claudin-1、Claudin-3的表达及意义

目的:探讨紧密连接蛋白claudin-1、claudin-3在子宫内膜异位症(内异症)发生发展中的作用及意义.方法:免疫组织化学方法检测正常子宫内膜、内异症患者的在位内膜和异位内膜组织中claudin-1、claudin-3蛋白质的表迭.结果:chudin-1、claudin-3蛋白主要定位于子宫内膜细胞的细胞膜/质,内异症患者的异位内膜中claudin-1、claudin-3蛋白的表达显著低于在位内膜及正常子宫内膜(P＜0.01).结论:claudin-1、claudin-3蛋白在内异症患者异住内膜中的表达明显下调,二者可能与内异症的发生发展相关.     

趋化因子CCL11-糖胺聚糖-趋化因子受体CCR3的相互作用研究

目的 探究趋化因子CCL11与受体CCR3、糖胺聚糖（glycosaminoglycans,GAGs）相互作用过程及机制,为深入阐明CCL11-GAGs-CCR3相互作用关系提供理论参考。方法 利用基因工程技术,构建筛选了CCR3-EGFP单分子表达水平的CHO稳转细胞系,利用全内反射荧光成像（total internal reflection fluorescence,TIRF）与等温滴定量热（isothermal titration calorimetry,ITC）技术研究了不同体外溶液条件下GAGs与CCL11的相互作用,并利用趋化实验及活细胞单分子成像实验考察了GAGs-CCL11对CCR3-EGFP稳转细胞趋化行为的调控及CCR3-EGFP在细胞膜上聚集状态的影响。结果 随着硫酸软骨素链长度的增加,其与CCL11结合放热增多,表明其相互作用力增强,其促进CCL11聚集作用增强。单分子荧光成像技术结合趋化试验研究发现,不同种类及不同比例的GAGs均会影响CCL11与CCR3的相互作用,GAGs的加入,抑制了CCL11对CCR3-EGFP稳转细胞的趋化效应及促CCR3-EGFP聚集的能力,且随着硫酸软骨素分子质量的增加,抑制作用显著增强。结论 GAGs的存在可以显著调控CCL11的聚集状态,进而影响其与受体CCR3的相互作用,本研究为进一步阐明CCL11-GAGs-CCR3相互作用关系提供了一定的实验基础。     

CXCL12/CXCR4/CXCR7轴在子宫内膜异位症中的研究进展

子宫内膜异位症(endometriosis, EMT)是常见的妇科疾病,发病率高,且有年轻化的趋势。因其治疗困难且复发率高,严重影响了女性的生活质量和生育能力。研究发现趋化因子CXCL12与其受体CXCR4和CXCR7在恶性肿瘤中起重要作用。虽然EMT为良性疾病,但有恶性肿瘤的生物学特征,近来发现CXCL12/CXCR4/CXCR7轴可以影响子宫内膜异位症的定植、侵袭和转移。本文就当前国内外研究CXCL12/CXCR4/CXCR7轴在EMT发生发展过程中的作用进行了综述,旨在为EMT的治疗找到新靶点。     

VEGF和IGF-I在子宫内膜异位症患者血清中的表达及临床意义

目的：探究血管内皮生长因子（VEGF）和胰岛素样生长因子-I（IGF-I）在子宫内膜异位症患者血清中的表达及临床意义,为子宫内膜异位症的治疗提供参考。方法：选取我院2015 年1 月至2016 年1 月收治的子宫内膜异位症患者50 例为实验组,另选体检中心健康妇女50 例为对照组。实验组患者根据疾病不同分期分为I、II期（n=24）和III、IV 期（n=26）。通过酶联免疫吸附法（ELISA）检测两组对象血清中VEGF和IGF-I的水平,采用Pearson相关分析法分析实验组患者血清中VEGF 和IGF-I 表达的相关性。结果：实验组患者血清中VEGF和IGF-I的水平均明显高于对照组,差异有统计学意义（P<0.05）;实验组III、IV期患者血清中VEGF和IGF-I的水平明显高于I、II期患者,差异有统计学意义（P<0.05）;Pearson 相关性分析显示实验组患者血清中VEGF和IGF-I的水平变化呈正相关关系（r=0.507,P<0.05）。结论：子宫内膜异位症患者血清中VEGF和IGF-I的水平高于正常水平,并随着病情的加重而不断升高,且二者呈正相关关系,可协同作用加快病情发展。     

Alkylsulfone-containing trisubstituted cyclohexanes as potent and bioavailable chemokine receptor 2 (CCR2) antagonists

We describe novel alkylsulfones as potent CCR2 antagonists with reduced hERG channel activity and improved pharmacokinetics over our previously described antagonists. Several of these new alkylsulfones have a profile that includes functional antagonism of CCR2, in vitro microsomal stability, and oral bioavailability. With this improved profile, we demonstrate that two of these antagonists, 2 and 12, are orally efficacious in an animal model of inflammatory recruitment.     

Hypoxia inhibits the expression of the CCR5 chemokine receptor in macrophages

Hypoxia, a decrease in oxygen tension occurring in pathological tissues, has a profound effect on macrophage functions. Here, we provide the first evidence that hypoxia inhibits CCR5 chemokine receptor expression in mouse macrophages. CCR5 was constitutively expressed in macrophages and upregulated by IFNgamma. Hypoxia downregulated both constitutive and IFNgamma-induced CCR5 mRNA and protein. Reoxygenation of hypoxic cells reverted CCR5 inhibition. CCR5 upregulation by IL-10, LPS, and IL-4 was also antagonized by hypoxia. CCR5 inhibition may be a way to retain/concentrate recruited macrophages at hypoxic sites or a feedback mechanism to control the autocrine activation of macrophages which produce CCR5 ligands.     

HIV chemokine receptor inhibitors as novel anti-HIV drugs

The chemokine receptors CXCR4 and CCR5 are the main coreceptors used by the T-cell-tropic (CXCR4-using, X4) and macrophage-tropic (CCR5-using, R5) HIV-1 strains, respectively, for entering their CD4⁺ target cells. In this review, we focus on the function of these chemokine receptors in HIV infection and their role as novel targets for viral inhibition. Besides some modified chemokines with antiviral activity, several low-molecular weight CCR5 and CXCR4 antagonistic compounds have been described with potent antiviral activity. The best CXCR4 antagonists described are the bicyclam derivatives, which consistently block X4 but also R5/X4 viral replication in PBMCs. We believe that chemokine receptor antagonists will become important new antiviral drugs to combat AIDS. Both CXCR4 and CCR5 chemokine receptor inhibitors will be needed in combination and even in combinations of antiviral drugs that also target other aspects of the HIV replication cycle to obtain optimum antiviral therapeutic effects.     

Interaction of soluble CD4 with the chemokine receptor CCR5

The chemokine receptor CCR5 is constitutively associated with the T cell co-receptor CD4 in plasma cell membranes. The CD4-CCR5 complex exhibits distinct binding properties for macrophage inflammatory protein 1beta (MIP-1beta) and enhanced G-protein signaling as compared with those of CCR5 alone. Here we report that recombinant soluble CD4, when refolded into its dimeric form, allosterically modulates CCR5 and decreases the affinity for its natural ligand MIP-1beta. Monomeric soluble CD4 had little inhibitory effect on CCR5. In contrast, the two-domain amino-terminal fragment of soluble CD4 was able to completely inhibit the interaction of CCR5 with MIP-1beta. Thus, we suggest that various conformational states of CD4 exist, which differ markedly with regard to inhibiting the interaction of CCR5 with its ligand MIP-1beta. R5-tropic HIV-1 glycoprotein 120, but not interleukin-16, the natural agonist, or X4-tropic glycoprotein 120, inhibited MIP-1beta binding to CCR5 in the presence of monomeric and dimeric soluble CD4.     

小鼠CCR7基因重组腺病毒的构建与鉴定

目的:构建含有小鼠趋化因子受体-7(CCR7)基因的重组腺病毒,为下一步转染不成熟树突状细胞(imDC)诱导免疫耐受研究奠定基础。方法:提取小鼠胸腺总RNA,应用逆转录PCR(RT-PCR)方法,以自行设计的带有酶切位点的引物,扩增获得CCR7基因全部序列,经过T-A克隆,酶切亚克隆到穿梭质粒pAdTrack-CMV上,在BJ5183菌内和pAdeasy-1同源重组,筛选阳性克隆,酶切鉴定,线性化后脂质体法转染HEK293细胞进行包装、PCR鉴定及扩增,得到含有CCR7基因的重组腺病毒,根据报告基因GFP测定病毒滴度。结果:成功构建小鼠CCR7基因重组腺病毒,病毒滴度为1×10~9U/mL。结论:该重组腺病毒载体的成功构建,为进一步研究携带CCR7基因的imDC的趋化迁移性等研究提供了一定的工作基础。     

Chemokine receptor CCR7 is expressed in muscle fibers in juvenile dermatomyositis

To better elucidate the pathogenesis of lymphocyte recruitment of memory CD4(+) T cells in inflammatory myopathies, we studied the expression of CCR5 and CCR7 on CD4 memory T cells in muscle tissue from 11 patients with juvenile dermatomyositis, six adult patients with polymyositis, two patients with Duchenne muscular dystrophy, and two patients with spinal muscular atrophy. A prevalent infiltration of CCR5(+) effector CD4 T memory cells is observed in inflammatory myopathies. Moreover, we found a strong expression of CCR7 in perifascicular atrophic and in degenerating/regenerating muscle fibers in juvenile dermatomyositis (JDM) but not in fibers from adult polymyositis and Duchenne muscular dystrophy. The selective expression of CCR7 in JDM may open new perspectives in the understanding of the pathogenesis of inflammatory myopathies, offering a new tool for the differential diagnosis of these disorders.     

Cell membrane hybrid bilayers containing the G-protein-coupled receptor CCR5

A hybrid bilayer membrane is a planar model membrane that is formed at an alkanethiol monolayer-coated gold surface by the spontaneous reorganization of phospholipid vesicles. Membrane vesicles from monkey kidney COS-1 cells also reorganize at an alkanethiol/lipid monolayer-coated surface resulting in the formation of a cell membrane hybrid bilayer. Atomic force microscopy and spectroscopic ellipsometry indicate that the cell membrane layer is equivalent to the thickness of one leaflet of the membrane and is continuous over large areas. Cell membrane hybrid bilayers were formed from membrane vesicles from COS-1 cells that were transiently transfected with a synthetic human CCR5 chemokine receptor gene. Preparations that contained "inside out" and "right side out" membrane vesicles were used. Binding of monoclonal antibodies to either the amino- or carboxyl-terminus of CCR5 was observed by surface plasmon resonance and confirmed the presence and the random orientation of these integral membrane receptors. Specific and concentration-dependent binding of the beta-chemokine RANTES to the cell membrane hybrid confirmed that CCR5 retained ligand-binding activity. The ability to form cell membrane hybrid bilayers that contain functional G-protein-coupled or other multispanning receptors without requiring protein isolation, purification, and reconstitution offers a promising method for the rapid screening of potential ligands.     

CCR7和B7-2在抗原负载树突状细胞诱导特异性CTL抗肿瘤效应中的表达和意义

目的:研究CCR7(趋化因子受体7)和B7-2(白细胞分化抗原86)与抗原负载树突状细胞(dentritic cell,DC)诱导特异性CTL(细胞毒性T淋巴细胞)抗肿瘤效应的关系.方法:分离和培养DC,制备B16黑色素瘤细胞抗原,进行共培养,即为抗原负载的DC,建立B16黑色素瘤小鼠模型,于肿瘤周围皮下注射抗原负载的DC.应用原位杂交和免疫组织化学方法检测CCR7和B7-2的表达情况.结果:原位杂交和免疫组织化学染色显示,CCR7和B7-2阳性细胞主要分布于肿瘤周围组织,随着注射抗原负载DC时间的进展,CCR7和B7-2呈强阳性表达.结论:CCR7和B7-2的表达与抗原负载树突状细胞诱导特异性CTL抗肿瘤效应有关.     

VEGF-C和CCR7的表达与卵巢癌淋巴结转移之间的关系

目的观察血管内皮生长因子(VEGF)-C和趋化因子受体CCR7在卵巢癌组织内的表达情况,分析VEGF-C和CCR7的表达与癌淋巴结转移之间的关系。方法取卵巢癌病例72例,其中,淋巴结转移组46例,无淋巴结转移组26例。应用免疫组化技术观察VEGF-C和CCR7在卵巢癌组织内的表达。结果 VEGF-C和CCR7主要表达于卵巢癌细胞胞浆或/和胞膜内,VEGF-C和CCR7在淋巴结转移组的阳性表达率分别是71.7%和78.2%,在无淋巴结转移组的表达率分别是30.8%和26.9%,二者在淋巴结转移组的表达率均明显高于无淋巴结转移组(P<0.01)。VEGF-C和CCR7蛋白同时阳性表达在淋巴结转移组和非淋巴结转移组中的表达率分别为65.2%和15.4%,VEGF-C和CCR7的表达具有显著的相关性(P<0.01),联合检测VEGF-C和CCR7诊断卵巢癌淋巴结转移具有较高的准确度,ROC曲线下面积达0.791。结论 VEGF-C和CCR7表达在卵巢癌淋巴结转移过程中发挥重要作用,VEGF-C和CCR7在促进卵巢癌淋巴结转移中可能具有一定的协同作用,二者联合检测有助于预示卵巢癌淋巴结转移的判断。