生物活性肽——蛋氨酸脑啡肽治疗白血病一例报告

蛋氨酸脑啡肽是内源性阿片样物质。它是由5个氨基酸残基组成的多肽,其第5位氨基酸残基为蛋氨酸。通过抽取病人静脉血,分离淋巴细胞,进行体外扩增激活,然后回输患者体内,患者外周血白细胞计数为3．6×10^9／L,血红蛋白122g/L,血小板116×10^9／L,异常淋巴细胞消失,幼粒细胞消失。骨髓象示粒细胞系统、红细胞系统增生活跃,各阶段细胞比值及形态大致正常;淋巴细胞比值减低,形态正常;造血功能恢复。由此可见,蛋氨酸脑啡肽体外激活免疫系统并回输体内作为一种新的治疗白血病的方法,具有广泛的应用前景。     

生物活性肽——蛋氨酸脑啡肽的免疫

源自肾上腺前脑啡肽原的具有吗啡样生物活性的内源性神经肽蛋氨酸脑啡肽(methionine enkephalin,MENK),由５个氨基酸残基Tyr Gly Gly Phe Met组成,与G蛋白偶联的７次跨膜特异性受体结合后发挥不同的生物学功能。目前,对蛋氨酸脑啡肽的作用及其机制研究已经取得了很大进展。本文就MENK对于免疫系统的调节及其对肿瘤、自身免疫病、艾滋病等免疫系统相关疾病的治疗作用予以综述。     

蛋氨酸脑啡肽质量标准研究

建立了蛋氨酸脑啡肽的质量标准。采用液相色谱-质谱联用仪检测蛋氨酸脑啡肽分子量和鉴别蛋氨酸脑啡肽的各氨基酸组成,反相高效液相色谱测定蛋氨酸脑啡肽含量。蛋氨酸脑啡肽分子量为573.7,氨基酸组成为Tyr-Gly-Gly-Phe-Met。蛋氨酸脑啡肽质量浓度在7~280 mg/L(r=0.999 8)内线性关系良好,平均回收率为98.54%,RSD为0.89%。所建立的方法科学,可靠,重复性好。可准确地对蛋氨酸脑啡肽进行定性定量检测。     

气相色谱法测定液相合成蛋氨酸脑啡肽中杂质残留量

测定液相法合成蛋氨酸脑啡肽中三氟乙酸和醋酸的残留含量。应用气相色谱法,程序升温,离子火焰反应检测器,定量测定。结果可见,该法线性范围为0.001～0.08 mg/L,在此浓度范围内与峰面积呈良好的线性关系,方法的检出限为0.0003～0.012 mg/L,测定的回收率为97.2%～101.7%,相对标准偏差<1.09%。所建立的检测方法简便,快速,灵敏度高,重复性好,可应用于检测蛋氨酸脑啡肽中残留醋酸的含量。     

生物活性肽—蛋氨酸脑啡肽免疫调节作用研究进展

蛋氨酸脑啡肽( methionine-enkephaIin,Met-ENK)是具有内源性吗啡样活性物质,它是由5个氨基酸残基组成的多肽,其第5位氨基酸残基为蛋氨酸.目前,对Met-ENK的生物学功能及其作用机制的研究已取得了显著进展.就Met-ENK对免疫细胞调控、抗肿瘤作用等方面做一综述.     

刺激大鼠尾壳核对脑内脑啡肽含量的影响

为确定刺激某一脑区时,远隔脑区脑啡肽含量是否改变;如有改变,甲-脑啡肽和亮-脑啡肽的变化是否一致。本工作在乌拉坦麻醉大鼠上,用放射免疫测定方法测定了电刺激尾壳核对六个脑区(下丘脑、纹状体、丘脑、海马、皮层及脑干)两种脑啡肽含量的影响。放射免疫测定的结果表明:(1)动物经乌拉坦麻校舍醉及手术操作后,除纹状体外,其它五个脑区亮-脑啡肽含量均明显降低,甲-脑啡肽含量只有下丘脑和丘脑明显降低,其它脑区无显著变化。(2)在乌拉坦麻醉及手术的基础上,再刺激尾壳核,除丘脑无显著变化外,其它五个脑区甲-脑啡肽含量均明显降低,亮-脑啡肽含量均明显升高,甲-脑啡肽/亮-脑啡肽比值减少(其中纹状体的比值改变无统计学显著性)。以上结果表明,刺激大鼠尾壳核可明显影响远隔脑区脑啡肽含量。关于甲-脑啡肽和亮-脑啡肽反向变化的意义有待进一步研究。     

S- 腺苷蛋氨酸对梗阻性黄疸患者术后肝功能及营养状况影响的临床研究

目的：探讨S-腺苷蛋氨酸对梗阻性黄疸患者术后肝功能及营养状况的影响。方法：选择2010 年8 月至2012年7 月我院肝 胆病区收治的90 例梗阻性黄疸患者为研究对象,随机分为S- 腺苷蛋氨酸治疗组（48 例）和对照组（42 例）,比较和分析静脉滴注 S-腺苷蛋氨酸对梗阻性黄疸患者术后第5 d、10 d肝功能及营养指标的影响。结果：术后5 d、10 d,两组患者血总胆红素、直接胆 红素、谷丙转氨酶、酌-谷氨酰转肽酶、碱性磷酸酶水平较术前1d 显著降低,且组内比较差异有统计学意义（P<0.05）,治疗组以上指 标的下降程度较对照组更明显,差异有统计学意义（P<0.05）。术后第10d,两组患者的血白蛋白、前白蛋白、转铁蛋白水平较术后 第5 d显著改善（P<0.05）;术后第5、10 d,两组组间血白蛋白、前白蛋白、转铁蛋白水平比较差异有统计学意义（P<0.05）。结论：梗 阻性黄疸患者术后应用腺苷蛋氨酸能促进黄疸消退,加快胆红素的排泄和肝功能的恢复,有利于患者营养状况的改善。     

S-腺苷蛋氨酸对梗阻性黄疸患者术后肝功能及营养状况影响的临床研究

目的：探讨S-腺苷蛋氨酸对梗阻性黄疸患者术后肝功能及营养状况的影响。方法：选择2010年8月至2012年7月我院肝胆病区收治的90例梗阻性黄疸患者为研究对象,随机分为S-腺苷蛋氨酸治疗组（48例）和对照组（42例）,比较和分析静脉滴注S-腺苷蛋氨酸对梗阻性黄疸患者术后第5d、10d肝功能及营养指标的影响。结果：术后5d、10d,两组患者血总胆红素、直接胆红素、谷丙转氨酶、1．谷氨酰转肽酶、碱性磷酸酶水平较术前1d显著降低,且组内比较差异有统计学意义（P〈0．05）,治疗组以上指标的下降程度较对照组更明显,差异有统计学意义（P〈0．05）。术后第10d,两组患者的血白蛋白、前白蛋白、转铁蛋白水平较术后第5d显著改善（P〈0．05）;术后第5、10d,两组组间血白蛋白、前白蛋白、转铁蛋白水平比较差异有统计学意义（P〈0．05）。结论：梗阻性黄疸患者术后应用腺苷蛋氨酸能促进黄疸消退,加快胆红素的排泄和肝功能的恢复,有利于患者营养状况的改善。     

胎脑黑质脑内移植矫正PD鼠纹状体内脑啡肽mRNA过度表达

用６－ＯＨＤＡ损毁大鼠一侧黑质—纹状体通路可引起ＰＤ模型鼠脑纹状体内多巴胺匿乏．同时,亦可导致脑啡肽ｍＲＮＡ过度表达。我们用地高辛标记的ｃＲＮＡ探针对纹状体内脑啡肽ｍＲＮＡ含量进行了斑点杂交定量研究,发现损伤侧脑啡肽ｍＲＮＡ含量较健侧增高８０％,胎中脑移植到失神经支配的纹状体内,脑啡肽ｍＲＮＡ过度表达得以矫正至正常水平,说明胎多巴胺能神经元脑内移植能够调节脑啡肽基因表达,提供了移植物能与宿主发生神经整合、建立突触联系的间接证据。     

S- 腺苷蛋氨酸在肝病患者中抗抑郁作用的研究进展

S-腺苷蛋氨酸(S-adenosyl-L-methionine,SAMe)是一种天然分子存在于人体所有细胞中,在肝脏、肾上腺以及松果体中具有较高浓度,在大脑中亦均匀分布。S-腺苷蛋氨酸通过抗氧化自由基及促进肝细胞再生等机制对肝细胞具有多重保护作用长久以来被广泛应用于肝病所致的肝内胆汁淤积、抗肝纤维化等治疗。近年来,研究亦发现S-腺苷蛋氨酸可增加患者脑中神经元膜的流动性及促进兴奋性神经递质的产生等对改善肝病患者情绪、治疗肝病患者抑郁症等方面具有重要的双重作用。通过静脉或口服用药,均可提高患者脑脊液中SAMe浓度,对于轻、中度抑郁患者的辅助治疗,更具安全性、有效性,这为人类的"生理-心理"疾病的治疗带来更广阔的应用前景。现将其对肝病患者抗抑郁的作用机制及临床应用综述如下。     

晚期胃癌患者化疗中胸腺肽α1治疗疗效的临床研究

目的：探讨应用流式细胞仪（FCM）检测胃癌患者外周血免疫指标,以评价胸腺肽α1联合化疗对胃癌患者免疫功能的影响。方法：70例胃癌患者随机分为用药组和对照组。用药组35例采用胸腺肽α1＋化疗,对照组35例单用化疗,两组化疗方案相同。胸腺肽α1每次1.6mg,每周2次,连续4周（共8针）。28天为一疗程,共用2个疗程。采用FCM检测两组患者化疗前后外周血CD4、CD8、CD3＋CD4＋、CD3＋CD8＋免疫指标。结果：用药组CD4、CD8、CD3＋CD4＋、CD4/CD8均高于化疗前和对照组化疗后水平,均具有统计学意义（P〈0.05）。结论：胸腺肽α1配合化疗能提高患者的免疫功能,用FCM检测外周血免疫指标为临床观察肿瘤患者的免疫状态提供有效的依据。     

TKI治疗晚期tPd,细胞肺癌患者免疫功能的变化及意义

目的：通过观察晚期非小细胞肺癌患者TKI治疗前后外周血IgG、[gM、IgA、C3、c4、C-反应蛋白及CD3＋、CD4＋、CD8＋、CD4＋／CD8＋细胞的表达变化,探讨TKI治疗对晚期非小细胞肺癌患者免疫功能的影响及意义。方法：检测TKI组30例非小细胞肺癌患者TKI治疗前、治疗一个月后外周血IgG、IgM、IgA、C3、C4、C-反应蛋白及CD3＋、CD4＋,CD8＋、CD4＋／CD8＋细胞表达水平,分析表达变化及与疗效的关系。30例非小细胞肺癌患者作为对照组。结果：治疗前,TKI组与对照组IgG、IgM、IgA、C3、C4、c＋反应蛋白水平基本正常,但CD4＋细胞数量减低、CD4＋／CD8＋比值较低、CD8＋细胞数量增高,两组相比IgG、IgM、IgA、C3、C4、C-反应蛋白、CD3＋、CD4＋．CD8＋、CD4＋／CD8‘差异均无统计学意义（P〉0．05）;TKI治疗一个月后,TKI组与对照组IgG、IgM、IgA、C3、C4、C-反应蛋白水平无明显变化,而CD4＋细胞数量增多、CD4＋／CD8＋较前增高,CD8＋细胞数量较前减低,两组相比CD3＋、IgG、IgM、IgA、C3、C4、c-反应蛋白差异无统计学意义（P〉0．05）,而CD4＋．CD4＋／CD8＋、CD8＋差异有统计学意义（P〈O．01）。结论：TKI治疗后,晚期非小细胞肺癌患者细胞免疫功能得到改善,体现在CD4＋,CD8＋细胞数量的变化上,且TKI治疗的疗效可通过比较外周血CD4＋、CD4＋／CD8＋、cD8＋细胞表达变化体现。     

TKI治疗晚期非小细胞肺癌患者免疫功能的变化及意义

目的:通过观察晚期非小细胞肺癌患者TKI治疗前后外周血IgG、IgM、IgA、C3、C4、C-反应蛋白及CD3+、CD4+、CD8+、CD4+/CD8+细胞的表达变化,探讨TKI治疗对晚期非小细胞肺癌患者免疫功能的影响及意义。方法:检测TKI组30例非小细胞肺癌患者TKI治疗前、治疗一个月后外周血IgG、IgM、IgA、C3、C4、C-反应蛋白及CD3+、CD4+、CD8+、CD4+/CD8+细胞表达水平,分析表达变化及与疗效的关系。30例非小细胞肺癌患者作为对照组。结果:治疗前,TKI组与对照组IgG、IgM、IgA、C3、C4、C-反应蛋白水平基本正常,但CD4+细胞数量减低、CD4+/CD8+比值较低、CD8+细胞数量增高,两组相比IgG、IgM、IgA、C3、C4、C-反应蛋白、CD3+、CD4+、CD8+、CD4+/CD8+差异均无统计学意义(P0.05);TKI治疗一个月后,TKI组与对照组IgG、IgM、IgA、C3、C4、C-反应蛋白水平无明显变化,而CD4+细胞数量增多、CD4+/CD8+较前增高,CD8+细胞数量较前减低,两组相比CD3+、IgG、IgM、IgA、C3、C4、C-反应蛋白差异无统计学意义(P0.05),而CD4+、CD4+/CD8+、CD8+差异有统计学意义(P0.01)。结论:TKI治疗后,晚期非小细胞肺癌患者细胞免疫功能得到改善,体现在CD4+、CD8+细胞数量的变化上,且TKI治疗的疗效可通过比较外周血CD4+、CD4+/CD8+、CD8+细胞表达变化体现。     

注射液胸腺法新（日达仙）辅助希罗达联合奥沙利铂（XELOX方案）治疗 晚期胃癌的临床疗效观察

目的:探讨注射液胸腺法新(日达仙)辅助希罗达联合奥沙利铂治疗晚期胃癌的临床疗效和对患者免疫功能及生活质量的影响。方法:选择2010-2013入院治疗的晚期胃癌患者116例,随机平均分为实验组(58例)和对照组(58例)。对照组给予卡培他滨片(希罗达)联合奥沙利铂,试验组在对照组的基础上辅用注射液胸腺法新(日达仙)治疗。每治疗两到三个周期后进行一次系统疗效评估,评估项目包括影像学腹部CT、胸片、腹部彩超、血常规、尿常规、肝功、肾功,肿瘤标记物癌胚抗原(CEA)、糖类抗原(CA19-9,CA72-4)、免疫指标CD3+、CD8+、CD4/CD8、NK细胞,总治疗周期为6-8个周期。结果:实验组疾病治疗有效率为48%,对照组有效率为29%,两组比较差异具有统计学意义(p=0.041),实验组患者中位无疾病进展期(PFS)为12.5月,显著高于对照组10.1月,两组比较具有统计学意义(P0.05)。实验组患者生活质量评分、外周血CD3+、CD8+、CD4/CD8、NK细胞数量均显著优于对照组(P0.05)。结论:注射液胸腺法新(日达仙)辅助希罗达联合奥沙利铂(XELOX方案)能够提高晚期胃癌的临床疗效,改善患者的生活质量和免疫力,减少患者化疗用药的毒副反应。     

乌苯美司联合紫杉醇与顺铂治疗晚期非小细胞肺癌的疗效及对患者免疫功能的影响

目的:探讨乌苯美司联合紫杉醇与顺铂(TP)方案治疗晚期非小细胞肺癌的临床疗效及对患者免疫功能的影响。方法:选择2013年10月-2015年6月在我院接受治疗的晚期非小细胞肺癌患者60例,随机分为研究组和对照组。两组患者均给予全身化疗治疗,研究组在此基础上给予口服乌苯美司治疗。观察并比较两组患者的临床疗效、免疫功能变化及不良反应的发生情况。结果:研究组患者治疗有效率明显高于对照组,差异具有统计学意义(P0.05);两组肿瘤控制情况比较,差异无统计学意义(P0.05);与治疗前比较,两组患者治疗后外周血CD3~+,CD4~+,CD4~+/CD8~+及NK细胞升高,而CD8~+下降,差异具有统计学意义(P0.05);与对照组比较,研究组患者治疗后外周血CD3~+,CD4~+,CD4~+/CD8~+及NK细胞显著升高,而CD8~+显著降低,差异具有统计学意义(P0.05);研究组患者白细胞减少、恶心、呕吐及骨髓抑制等不良反应的发生率低于对照组,差异具有统计学意义(P0.05)。结论:乌苯美司联合TP方案能够改善晚期非小细胞肺癌患者机体免疫功能,减少不良反应的发生率,值得临床推广应用。     

Effect of adoptive T-cell immunotherapy on immunological parameters and prognosis in patients with advanced pancreatic cancer

Background aimsImmunotherapy is effective for many types of cancer, but its benefits in advanced pancreatic cancer, which has a poor prognosis, are not well established. In this study, the authors examined the effects of adoptive T-cell immunotherapy (ATI) on immune cell profiles and prognosis in patients with unresectable advanced pancreatic cancer.MethodsSeventy-seven patients with unresectable advanced pancreatic cancer were treated with six cycles of αβ T cells alone or in combination with chemotherapy or chemoradiation. Immune cell profiles in peripheral blood samples obtained before and after treatment were comprehensively evaluated by flow cytometry. Furthermore, associations between changes in immune cell frequencies and prognosis were determined.ResultsATI prolonged survival to 18.7 months compared with previous estimates of 6.2–11.1 months for patients treated with chemotherapy alone. ATI decreased CD3+CD4+CD8? T cell frequency in peripheral blood and increased CD3+CD4?CD8+ T cell frequency. An increase in CD3+ T cells and CD3+TCRγδ? T cells in peripheral blood after treatment was associated with a good prognosis.ConclusionsATI altered the immune profile in peripheral blood, including CD3+CD4?CD8+ T cells, and improved prognosis in pancreatic cancer.     

Treatment combinations targeting apoptosis to improve immunotherapy of melanoma

Immunotherapy based on T cell responses to the tumor is believed to involve killing of cancer cells by induction of apoptosis. The predominant mechanisms are death ligand-induced signaling mainly by TNF-related apoptosis-inducing ligand (TRAIL) mediated by CD4 T cells, monocytes and dendritic cells, and perforin/granzyme mediated apoptosis mediated by CD8 T cells and NK cells. Resistance against TRAIL involves loss of TRAIL death receptors and/or activation of the MEK and/or Akt signal pathways. Resistance to CD8 CTL responses also involves activation of the MEK and/or Akt pathways. Apoptosis induced by immune responses is regulated by the Bcl-2 family of proteins. Many reagents have been developed against the Bcl-2 antiapoptotic proteins and clinical trials combining them with immunotherapy are awaited. The second group of agents that regulate the Bcl-2 family of proteins are the signal pathway inhibitors. Clinical trials with inhibitors of RAS, RAF or MEK are in progress and would appear an exciting combination with immunotherapy. One of the main drivers of resistance to apoptosis are adaptive mechanisms that allow cancer cells to overcome endoplasmic reticulum (ER) stress. These adaptive mechanisms inhibit practically all known apoptotic pathways and create an acidic environment that may reduce infiltration of lymphocytes against the tumor. The signal pathway inhibitors may be effective against these adaptive processes but additional agents that target ER stress pathways are in development. In conclusion, combination of immunotherapy with agents that target antiapoptotic mechanisms in cancer cells offers a new approach that requires evaluation in clinical trials.     

EC144, a synthetic inhibitor of heat shock protein 90, blocks innate and adaptive immune responses in models of inflammation and autoimmunity

Heat shock protein 90 (Hsp90) is a molecular chaperone involved in folding and stabilizing multiple intracellular proteins that have roles in cell activation and proliferation. Many Hsp90 client proteins in tumor cells are mutated or overexpressed oncogenic proteins driving cancer cell growth, leading to the acceptance of Hsp90 as a potential therapeutic target for cancer. Because several signal transduction molecules that are dependent on Hsp90 function are also involved in activation of innate and adaptive cells of the immune system, we investigated the mechanism by which inhibiting Hsp90 leads to therapeutic efficacy in rodent models of inflammation and autoimmunity. EC144, a synthetic Hsp90 inhibitor, blocked LPS-induced TLR4 signaling in RAW 264.7 cells by inhibiting activation of ERK1/2, MEK1/2, JNK, and p38 MAPK but not NF-κB. Ex vivo LPS-stimulated CD11b(+) peritoneal exudate cells from EC144-treated mice were blocked from phosphorylating tumor progression locus 2, MEK1/2, and ERK1/2. Consequently, EC144-treated mice were resistant to LPS administration and had suppressed systemic TNF-α release. Inhibiting Hsp90 also blocked in vitro CD4(+) T cell proliferation in mouse and human MLRs. In vivo, semitherapeutic administration of EC144 blocked disease development in rat collagen-induced arthritis by suppressing the inflammatory response. In a mouse collagen-induced arthritis model, EC144 also suppressed disease development, which correlated with a suppressed Ag-specific Ab response and a block in activation of Ag-specific CD4(+) T cells. Our results describe mechanisms by which blocking Hsp90 function may be applicable to treatment of autoimmune diseases involving inflammation and activation of the adaptive immune response.     

氟比洛芬酯对食管癌患者围术期外周血淋巴细胞亚群的影响

目的:探究氟比洛芬酯对食管癌患者围术期外周血淋巴细胞亚群的影响。方法:选择2014年6月~2016年8月期间在我院择期行食管癌根治术患者72例为研究对象,采用随机数字法将其分为氟比洛芬酯组(39例)和对照组(33例),患者均给予常规麻醉处理,对照组患者泵入5 mg托烷司琼与20μg/kg芬太尼;氟比洛芬酯组患者给予氟比洛芬酯2 mg/kg。评价术后12 h、24 h和48 h患者疼痛情况(VAS评分),并于术前1 h、术后24 h、术后72 h检测患者血清T细胞中CD3~+、CD4~+、CD8~+及CD56~+比例。结果:两组患者术后不同时刻VAS评分比较,差异均无统计学意义(P0.05);术后24 h两组患者CD3~+、CD4~+、CD4~+/CD8~+均显著降低(P0.05),术后72 h两组患者CD3~+、CD4~+、CD4~+/CD8~+水平均升高,且氟比洛芬酯组患者恢复到术后1 h水平,而对照组患者均仍低于术后1 h,且术后72 h氟比洛芬酯组CD3~+、CD4~+、CD4~+/CD8~+均高于对照组,差异均有统计学意义(P0.05),而CD8~+与CD56~+比例在两组各个时点均没有变化(P0.05);两组患者均未发生严重不良反应。结论:食管癌患者在手术过程中均出现免疫抑制,氟比洛芬酯麻醉效果较好,且对机体免疫功能具有保护作用,促进手术患者免疫功能的恢复,具有重要的临床价值。     

Alterations of pineal gland and of T lymphocyte subsets in metastatic cancer patients: preliminary results

Melatonin (MLT), the main hormone produced by the pineal gland, has been seen to play a role in antineoplastic activity either by exerting a direct inhibitory effect on cancer cell growth, or by stimulating the immune system. Moreover, MLT blood levels have been shown to be often increased in cancer patients. On the basis of these data, a study was started to evaluate what relation exists between MLT levels and T lymphocyte subsets in patients with metastatic solid neoplasm. The study included 28 patients (breast: 10; non-small cell lung: 18). None of the patients was previously treated for their metastatic disease. Abnormally high MLT levels and a low T helper/suppressor ratio (CD4/CD8) were seen in 10/28 and in 11/28 patients, respectively. Serum mean levels of MLT were significantly higher in patients with low CD4/CD8 ratio than in those with a normal ratio. These results would suggest that immune dysfunctions may represent a signal for MLT release from the pineal in patients with metastatic solid neoplasm.