幽门螺杆菌感染与胃癌共刺激分子OX40和 OX40L表达的相关性

目的研究幽门螺杆菌(H.pylori)感染与胃癌共刺激分子OX40、OX40L表达的相关性。方法收集本院2017年1月至2019年1月进行胃癌手术的68例患者为研究对象。采用Giemsa染色和PCR方法检测胃癌组织H.pylori感染情况。应用免疫组织化学法检测胃癌和癌旁组织中共刺激分子OX40、OX40L的表达,并分析共刺激分子OX40、OX40L表达与胃癌组织H.pylori感染的相关性。分析OX40、OX40L表达与胃癌患者临床病理特征的关系。采用二元Logistic回归分析胃癌组织H.pylori感染的影响因素。结果胃癌组织中OX40、OX40L阳性表达率显著高于癌旁组织(均P0.05);Giemsa染色和PCR检测胃癌组织中H.pylori感染阳性率分别为86.76%(59/68)和91.18%(62/68)。H.pylori阳性胃癌组织中OX40、OX40L阳性表达率显著高于H.pylori阴性胃癌组织(均P0.05)。OX40、OX40L表达与胃癌患者肿瘤浸润深度、分化程度、T分期、淋巴结转移均无显著相关性(均P0.05),而与胃癌肿块大小相关(P0.05)。OX40、OX40L阳性表达是胃癌组织中H.pylori感染的危险因素。结论 H.pylori感染可能与胃癌组织中OX40、OX40L异常表达相关,可为探究H.pylori致癌机制提供一定参考。     

辛伐他汀对IFN-γ 诱导的人外周血单个核细胞OX40L 表达的影响

目的:探讨经IFN-γ刺激后,人外周血单个核细胞OX40L表达的变化,以及辛伐他汀对单个核细胞OX40L表达的影响.方法:将实验标本随机分为2组,分别干扰素-γ(IFN-γ)刺激组、辛伐他汀干预组.应用RT-PCR及Western blotting技术,观察IFN-γ诱导的人外周血单个核细胞OX40L表达情况及辛伐他汀对人单个核细胞OX40L表达的影响.结果:1.1000U/mlIFN-与人单个核细胞共同培养24h后,OX40L mRNA和蛋白水平的表达明显增加.2.预先给予10 mol/L的辛伐他汀干预1h可以明显降低IFN-诱导的OX40L表达.结论:IFN-可诱导人单个核细胞OX40L表达.辛伐他汀可以抑制IFN-诱导人单个核细胞OX40L表达的增强,从而可能抑制了OX40L信号通路介导的与炎症有关的血管损伤,延缓动脉粥样硬化的进程.     

辛伐他汀对IFN-y诱导的人外周血单个核细胞OX40L表达的影响

目的：探讨经IFN-y刺激后,人外周血单个核细胞OX40L表达的变化,以及辛伐他汀对单个核细胞OX40L表达的影响。方法：将实验标本随机分为2组,分别干扰素-y（IFN-y）刺激组、辛伐他汀干预组。应用RT—PCR及Western blotting技术,观察IFN-y诱导的人外周血单个核细胞OX40L表达情况及辛伐他汀对人单个核细胞OX40L表达的影响。结果：1．1000U／ml IFN-与人单个核细胞共同培养24h后,OX40LmRNA和蛋白水平的表达明显增加。2．预先给予10mol／L的辛伐他汀干预1h可以明显降低IFN-诱导的OX40L表达。结论：IFN-可诱导人单个核细胞OX40L表达。辛伐他汀可以抑制IFN-诱导人单个核细胞OX40L表达的增强,从而可能抑制了OX40L信号通路介导的与炎症有关的血管损伤,延缓动脉粥样硬化的进程。     

冠心病患者介入治疗前后血清IL-18、sCD40L 和hs-CRP 水平的 变化及其意义

目的:探讨冠心病患者血清白介素18(Interleukin-18,IL-18)、白细胞分化抗原40配体(CD40L)、及高敏C反应蛋白(hs-CRP)水平在经皮冠状动脉介入术治疗前后的变化和意义。方法:选择经冠状动脉造影确诊的冠心病患者85例,根据病变程度分为单支病变组(n=32)、双支病变组(n=28)和多支病变组(n=25),采用双抗体夹心ELISA法测定PCI术前术后血清IL-18、CD40L和hs-CRP水平。结果:血清IL-18水平测定结果:多支病变组高于双支病变组,双支病变组高于单支病变组;支架置入术后显著高于术前,差异均有统计学意义(P<0.01)。血清CD40L水平测定结果:多支病变组高于双支病变组和单支病变组,差异均有统计学意义(P<0.01),双支病变组与单支病变组间差异无统计学意义(P>0.05);支架置入术后较术前显著升高,差异有统计学意义(P<0.01)。血清hs-CRP水平测定结果:多支病变组高于双支病变组,双支病变组高于单支病变组;支架置入术后显著高于术前,差异均有统计学意义(P<0.01)。结论:冠心病患者血清IL-18、CD40L和hs-CRP与冠脉病变程度密切相关,介入治疗可使冠心病患者血清IL-18、CD40L和hs-CRP水平升高,监测血清中IL-18、CD40L和hs-CRP水平变化可了解治疗效果和炎症程度。     

不同剂量瑞舒伐他汀对老年冠心病患者疗效及安全性观察

目的:探讨不同剂量的瑞舒伐他汀对老年冠心病患者的疗效及安全性的影响。方法:抽取2012年6月至2013年6月于我院住院行冠脉造影检查示冠状动脉临界病变,诊断为冠状动脉粥样硬化性心脏病的患者210例(≥75岁),随机分为甲组,乙组,对甲组给予10 mg/d的瑞舒伐他汀药物口服治疗,乙组给予20 mg/d的瑞舒伐他汀药物口服治疗,其它协同药物服用方法均采集。收集患者开始服用瑞舒伐他汀药物前及服用后1月、6月、12月的血生化指标及颈动脉彩超结果,包括低密度脂蛋白胆固醇(Low-density Lipoprotein Cholesterol,LDL-C),总胆固醇(Total Cholesterol,TC),甘油三脂(Triglyceride,TG),高密度脂蛋白胆固醇(High-density Lipoprotein Cholesterol,HDL-C),超敏C反应蛋白(High Sensitivity C Reactive Protein,hs-CRP),颈动脉内-中膜厚度(Intima-media thickness,IMT),谷丙转氨酶(Alanine-Aminotransferase,ALT),观察这些指标服药前后变化的情况。结果:瑞舒伐他汀药物治疗1月、6月、12月后两组患者指标相比较,乙组患者较甲组患者TC、TG、LDL-C、hs-CRP、IMT降低更明显,而HDL-C升高水平则明显高于甲组,差别有统计学意义(P<0.05)。两组患者之间血生化指标ALT及药物治疗期间发生瑞舒伐他汀相关严重不良反应例数,无明显差别(P>0.05)。两组均无因心脏不良事件(MACE事件)而终止实验的患者。结论:对于老年冠心病患者,瑞舒伐他汀药物治疗可以获益,高强度的瑞舒伐他汀药物获益更显著,并且是安全的。     

瑞舒伐他汀联合丹参酮对老年心力衰竭患者血清NF-κB和IL-1β水平的影响

目的:探讨瑞舒伐他汀联合丹参酮对老年心力衰竭患者血清核转录因子kappa B(NF-κB)和白细胞介素-1β(IL-1β)水平的影响。方法:选择2011年7月~2015年7月于我院进行治疗的老年心力衰竭患者68例,按照电脑生成的随机数字表将所有患者随机分为实验组和对照组,每组各34例,对照组使用瑞舒伐他汀进行治疗,实验组患者在对照组的治疗基础上加用丹参酮进行治疗,对比分析两组患者血清核因子-κB(NF-κB)、白细胞介素-1β(IL-1β)、超敏C反应蛋白(hs-CRP)、一氧化氮(NO)、内皮素-1(ET-1)及B型钠尿肽(BNP)水平,并对两组患者的临床疗效进行评价。结果:与治疗前相比,治疗后两组患者血清NF-κB、IL-1β,ET-1、BNP及hs-CRP水平均降低,血清NO水平均升高(P0.05),且与对照组相比,实验组患者血清NF-κB、IL-1β、hs-CRP、ET-1及BNP水平较低,NO水平及临床总有效率较高(P0.05)。结论:瑞舒伐他汀联合丹参酮对老年心力衰竭具有很好的治疗效果,推测其机制与降低血清NF-κB、IL-1β、hs-CRP、ET-1及BNP水平及升高NO水平有关。     

瑞舒伐他汀和阿托伐他汀在急性冠脉综合症中对氯吡格雷抗血 小板活性的对比研究

目的：在急性冠脉综合征( acute coronary syndromes, ACS )的治疗中,抗血小板治疗及调脂治疗是最基础的治疗方案。近来 有学者提出,氯吡格雷和他汀类药物都经过细胞色素CYP 3A4 途径代谢,二者因存在竞争性抑制,有可能降低氯吡格雷抗血小板 的活性。本试验将针对阿托伐他汀及瑞舒伐他汀进行研究。方法：选择急性冠脉综合症的患者42 例,所有患者均接受氯吡格雷治 疗（负荷剂量300 mg,维持剂量75 mg/d）。随机分配为A、B 两组,A 组（n=20）服用阿托伐他汀治疗（20 mg/d）,B 组（n=22 服用瑞 舒伐他汀治疗（10 mg/d）。分别于氯吡格雷服用前、服药治疗后3 天、服药治疗后7 天后采静脉血送检,测定ADP（10 滋mol/L）诱导 的血小板聚集率。结果：阿托伐他汀组（A 组）及瑞舒伐他汀组（B 组）相比,服用氯吡格雷前ADP 诱导的血小板聚集率基线值无 统计学差异。服用氯吡格雷3 日及7 日后,ADP诱导的血小板聚集率明显降低,(3.85± 2.58)vs(3.09± 2.27),(0.65± 0.88)vs(1.05± 0.95),P>0.05,无明显统计学差异。结论：氯吡格雷的确可以降低血小板的活性。同时,短期之内氯吡格雷的抗血小板活性未受到 他汀类的影响,包括经过CPY3A4途径的他汀,如阿托伐他汀。     

瑞舒伐他汀对高血压合并高胆固醇血症患者血清高敏C 反应蛋白水平的影响*

摘要目的：观察瑞舒伐他汀对高血压合并高胆固醇血症患者血清高敏C 反应蛋白(hi-sensitive C-reactive protein,hs-CRP)水平的 影响。方法：选择60 例高血压合并高胆固醇血症的患者为研究对象,将其随机分为瑞舒伐他汀治疗组(实验组)和常规方法治疗组 (对照组),对照组仅给予常规治疗,实验组在常规组治疗的基础上每日加服一次瑞舒伐他汀10mg,疗程8 周,测定和比较两组患 者治疗前后血压、血清总胆固醇(Total cholesterol,TC)、低密度脂蛋白胆固醇(Low density lipoprotein-cholesterol,LDL-C)和hs-CRP 的水平。结果：治疗8 周后,实验组患者的血清SBP、DBP、TC、LDL-C、hs-CRP水平均较治疗前明显降低(P<0.05),且较对照组组更 低(P<0.05)。结论：瑞舒伐他汀辅助治疗能显著降低高血压合并高胆固醇血症患者的血压和血清血脂水平,并可减少血清高敏C 反应蛋白水平,有益于改善患者的预后。     

不同剂量瑞舒伐他汀对急性冠脉综合征患者炎症反应及预后的影响

目的:探讨不同剂量瑞舒伐他汀对急性冠脉综合征患者(ACS)炎症反应及预后的影响。方法:选择2014年5月至2015年10月间收治的120例ACS患者为研究对象,随机分为A、B、C三组,每组40例。所有患者先给予常规治疗,A组患者给予20mg/d普伐他汀治疗;B组患者给予10 mg/d瑞舒伐他汀;C组患者给予20 mg/d瑞舒伐他汀,3组均持续治疗6个月。比较3组患者治疗前后血脂、超敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)的水平;随访6个月心血管事件发生情况。结果:3组患者治疗后hs-CRP、TNF-α、IL-6水平均明显下降,且C组下降最明显,其次为B组,比较差异均有统计学意义(P0.05);治疗后血脂水平均的改善,且C组改善最明显,其次是B组,比较差异均有统计学意义(P0.05);随访6个月,C组心血管事件发生率为2.5%,远低于B组(10.0%)及A组(20.0%),差异具有显著性(P0.05)。结论:大剂量(20 mg/d)瑞舒伐他汀治疗ACS降脂、抗炎效果明显,预后相对较好,值得临床推广使用。     

冠心病患者血清sCD40L水平及其与hs-CRP关系研究

目的:研究冠心病患者血清sCD40L和hs-CRP临床特点及其关系,探讨其在冠心病预测和治疗中的意义.方法:采用夹心法酶联免疫吸附测定分析法及微粒子增强透射免疫分析法分别对对照组15例、稳定型心绞痛(SAP)组27例、不稳定型心绞痛(UAP)组35例及急性心肌梗死(AMI)组14例受试者血清sCD40L和hs-CRP水平进行检测,并观察其与冠脉狭窄程度的相关性.结果:1)、血清hs-CRP水平:SAP组、UAP组、AMI组呈依次递增(AMI组比UAP组、UAP组比SAP组P＜0.05,AMI组、UAP组比对照组(P＜0.01);SAP组与对照组血清hs-CRP水平相似;2)、血清sCIMOL水平:AMI组及UAP组血清sCD40L水平高于SAP组和对照组(P＜0.01),AMI组与UAP组之间、SAP与对照组间没有统计学差异;3)、相关分析显示血清sCD40L水平与hs-CRP水平显著相关(r=0.787,P＜0.0001),两者均与冠脉狭窄程度无相关性.结论:血清hs-CRP、sCD40L水平升高与冠心病临床表现类型和病情是否稳定有关,与冠状动脉狭窄程度无关,此两项指标能够用于冠心病病情不稳定性的判断和预测.     

免疫共刺激分子OX40L对乙型肝炎核酸疫苗的免疫佐剂作用

[目的]为了进一步增强HBV DNA疫苗的免疫反应,本研究将共刺激分子OX40L 作为HBV DNA疫苗的分子佐剂免疫小鼠,旨在探讨共刺激分子OX40L对HBV DNA疫苗诱导体液和细胞免疫应答的影响.[方法]我们将HBV DNA疫苗(pcDS2)单独或联合共刺激分子质粒pOX40L免疫C57BL/6小鼠;分别在第0,2,4周进行免疫,在第6周检测抗-HBs IgG、IgG1和IgG2a,T淋巴细胞增殖指数,细胞因子表达水平和体内细胞毒性T淋巴细胞杀伤作用(CTL)等免疫学指标.[结果]pceDS2联合pOX40L免疫组小鼠的抗-HBs水平显著提高,抗-HBs IgG亚类以IgG2a占优;免疫小鼠的T淋巴细胞体外经乙型肝炎表面抗原(HBsAg)刺激后,联合免疫组刺激指数(SI)明显高于pcDS2组;联合免疫组CD4 + T淋巴细胞的IL-4和IFN-γ表达水平及CD8 + T淋巴细胞的IFN-γ表达水平显著升高;DNA疫苗免疫的各组小鼠,HBsAg特异性体内CTL高于对照组,其中联合免疫组小鼠的体内CTL杀伤作用最强.[结论]共刺激分子OX40L不仅能增强HBV DNA疫苗诱导特异性体液免疫应答,还能增强特异性细胞免疫反应,尤其增强体内CTL的杀伤活性,为HBV DNA疫苗的研究奠定了基础.     

携带OX40L重组溶瘤病毒的构建及靶向杀伤肝癌的研究

目的 拯救携带OX40L的重组溶瘤流感病毒株，全面鉴定并评价其靶向杀伤肝癌细胞的效果。方法 将编码OX40L的基因片段嵌合在A/PuertoRico/8/34（PR8）的NS片段特定位置，利用反向遗传学技术（reverse genetics，RG），与PR8流感病毒的剩余7个骨架质粒pHW191-PB2、pHW192-PB1、pHW193-PA、pHW194-HA、pHW195-NP、pHW196-NA、pHW197-M共转染COSⅠ/MDCK细胞，成功拯救获得重组溶瘤流感病毒，命名为rFlu-OX40L。经血凝、TCID₅₀方法测定病毒滴度；重组病毒纯化后电镜观察病毒形态特征及大小分布；检测MDCK细胞病毒生长曲线；MTS法检测重组病毒对肝癌细胞活力的影响；流式细胞术检测重组病毒诱导肝癌细胞死亡方式；利用肝癌荷瘤小鼠模型评价重组病毒rFlu-OX40L在动物体内的抗肿瘤效果。结果 重组溶瘤流感病毒rFlu-OX40L可在鸡胚中稳定传代，HA效价达2^7~8，病毒滴度可达 7~8 LgTCID₅₀/ml；重组病毒rFlu-OX40L与流感病毒PR8生长曲线相一致，72 h达高峰；MTS结果显示，3 MOI重组病毒可显著降低肝癌细胞活力，且对正常肝细胞无明显影响，呈时间和剂量依赖性；流式细胞结果显示，重组病毒可显著诱导肝癌细胞凋亡，呈时间和剂量依赖性；利用肝癌荷瘤小鼠模型，瘤内注射重组溶瘤流感病毒rFlu-OX40L较PR8、PBS对照组，小鼠脾细胞中CD3+、CD4+、CD8+、CD45+、CD69+ T细胞数量呈现显著升高趋势。结论 携带OX40L的重组溶瘤流感病毒rFlu-OX40L在体内外均可选择性杀伤肝癌细胞，有望为肝癌的临床治疗提供新的免疫治疗策略。     

Soluble OX40L favors tumor rejection in CT26 colon carcinoma model

OX40 receptor-expressing regulatory T cells (Tregs) populate tumors and suppress a variety of immune cells, posing a major obstacle for cancer immunotherapy. Different ways to functionally inactivate Tregs by triggering OX40 receptor have been suggested, including anti-OX40 antibodies and Fc:OX40L fusion proteins. To investigate whether the soluble extracellular domain of OX40L (OX40Lexo) is sufficient to enhance antitumor immune response, we generated an OX40Lexo-expressing CT26 colon carcinoma cell line and studied its tumorigenicity in immunocompetent BALB/c and T cell deficient nu/nu mice.We found that soluble OX40L expressed in CT26 colon carcinoma favors the induction of an antitumor response which is not limited just to cells co-expressing EGFP as an antigenic determinant, but also eliminates CT26 cells expressing another fluorescent protein, KillerRed. Tumor rejection required the presence of T lymphocytes, as indicated by the unhampered tumor growth in nu/nu mice. Subsequent re-challenge of tumor-free BALB/c mice with CT26 EGFP cells resulted in no tumor growth, which is indicative of the formation of immunological memory. Adoptive transfer of splenocytes from mice that successfully rejected CT26 OX40Lexo EGFP tumors to naïve mice conferred 100% resistance to subsequent challenge with the CT26 EGFP tumor.     

Induction of xenogeneic islet transplantation tolerance by simultaneously blocking CD28-B7 and OX40-OX40L co-stimulatory pathways

It has been demonstrated that prolonged graft survival can be achieved through inhibiting the activation of T cells, and addition of soluble CTLA4Ig and OX40Ig proteins to mixed lymphocyte reactions can effectively inhibit T cell proliferation. To explore the potential of this type of treatment in xenotransplantation, we infected streptozotocin-induced diabetic BalB/c mice (H-2d) (200 mg/kg, IV) with 5×10⁸ pfu AdCTLA4Ig-IRES-OX40Ig on day 1 before islets transplantation through the tail vein. The results showed that this treatment prolonged the islet xenografts survival significantly. The reaction to exogenous glucose stimulation was normal and the cytokine secretion of the type Th1 cells was inhibited. The AdCTLA4Ig-IRES-OX40Ig-mediated treatment effectively induced the T cells into anergy and the Th1/Th2 cells into deviation. These results strongly supported the therapeutic potential of blockade of costimulation by Ad-CTLA4Ig-IRES-OX40Ig genes transfer in inducing the organ transplantation tolerance.     

Induction of xenogeneic islet transplantation tolerance by simultaneously blocking CD28-B7 and OX40-OX40L co-stimulatory pathways

Type 1 diabetes mellitus is an autoimmune disease caused by T cell-mediated destruction of pancreatic beta islets. With its incidence continuous rising in the pediatric age group in recent years, it is becoming a serious threat to the human health. Up to now, there has been no effective therapy for diabetes mellitus, and islet transplantation still remains a promising ap-proach to the treatment of Type 1 diabetes. However, two serious problems hinder the successful islets transplantation, tha…     

The expression of CD40-CD40L and activities of matrix metalloproteinases in atherosclerotic rats

This study investigated the expression of CD40, CD40 ligand (CD40L) and matrix metalloproteinases (MMPs) in dietary-induced atherosclerosis in rats. Wister rats were fed with high cholesterol diet (As group, n = 6) or with normal diet (N group, n = 6). Blood cells that express CD40 and CD40L were sorted by flow cytometry, the MMP-2 and MMP-9 were measured by zymography method. The morphological locations of MMP-2 and MMP-9 in the aorta were studied with immunohistochemistry and by microscopy. The results showed that the expression of CD40, CD40L and matrix metalloproteinase were higher in As group than those in control group. The MMP-2 and MMP-9 were positive in As group but negative in control group by immunohistochemistry study. Our results suggest that the expression of CD40 and CD40L in the blood cells and the activities of MMP-2 and MMP-9 in plasma were higher in As group than those in Normal group, indicating that they may contribute to the formation of atherosclerosis.     

OX40–OX40 ligand interaction may activate phospholipase C signal transduction pathway in human umbilical vein endothelial cells

Studies have recently supported the emerging role of OX40/OX40L interaction in atherosclerosis. The mechanism of OX40/OX40L interaction may be related to a variety of signal pathways. The most important signal pathway involves the activation of phospholipase C (PLC) which induces diacylglycerol–protein kinase C (DAG–PKC) and the inositol trisphosphate (IP₃)–intracellular free calcium ([Ca²⁺]_i) pathway. The aim of this work was to investigate whether OX40–OX40L interaction can stimulate the PLC signal pathway in human umbilical vein endothelial cells (HUVEC). The DAG and IP₃ level in HUVEC were measured by radio-enzymatic assay. The activity of PKC was detected by its ability to transfer phosphate from [γ-³²P]ATP to lysine-rich histone. [Ca²⁺]_i concentrations were measured by flow cytometric analysis. Results showed that the DAG level was markedly increased in a concentration-dependent, biphasic manner in HUVEC induced by OX40. The early phase was rapid, peaking at 30 s. The late phase reached the maximum level at 15 min and decayed slowly. OX40 increased PKC activity in a dose-dependent manner with two peaks at 40–50 s and 12–16 min, then decreased slowly, yet maintained a high level for at least 30 min. PKC activity was mainly in cytosol at rest and translocated from cytosol to membrane when stimulated by OX40. Similarly, OX40-induced rapid IP₃ formation coincided with the peak of DAG level. Moreover, OX40 also induced peak [Ca²⁺]_i responses including the rapid transient phase and the sustained phase. Anti-OX40L antibody significantly suppressed OX40-induced DAG–PKC and IP₃–[Ca²⁺]_i signal pathway activation in HUVEC. In conclusion, the data suggested that OX40–OX40L interaction induced a robust stimulation of phospholipase C signal transduction pathway in HUVEC.     

Basophil-associated OX40 Ligand Participates in the Initiation of Th2 Responses during Airway Inflammation

Asthma is characterized by increased airway submucosal infiltration of T helper (Th) cells and myeloid cells that co-conspire to sustain a chronic inflammation. While recent studies have demonstrated that the myeloid basophils promote Th2 cells in response to various types of allergens, the underlying mechanisms are poorly understood. Here, we found for the first time that in a mouse model of allergic asthma basophils highly expressed OX40 ligand (OX40L) after activation. Interestingly, blockade of OX40-OX40L interaction suppressed basophils-primed Th2 cell differentiation in vitro and ameliorated ovalbumin (OVA)-induced allergic eosinophilic inflammation mediated by Th2 activation. In accordance, the adoptive transfer of basophils derived from mediastinal lymph nodes (MLN) of OVA-immunized mice triggered a robust Th2 response and eosinophilic inflammation in wild-type mice but largely muted in OX40^−/− mice and mice receiving OX40L-blocked basophils. Taken together, our results reveal a critical role of OX40L presented by the activated basophils to initiate Th2 responses in an allergic asthma model, implicating OX40-OX40L signaling as a potential therapeutic target in the treatment of allergic airway inflammation.