急性白血病患者外周血Treg细胞与Th17细胞的相关性

目的:检测急性白血病患者外周血中Treg细胞与Th17细胞的比例以及相关细胞因子如IL-17、IL-6、TGF-β的变化,分析其相关性。方法:选取兰州大学第二医院血液科急性白血病初诊患者15例,另取15名健康志愿者为对照。流式细胞术检测外周血中CD3+CD4+TIL-17+辅助性T细胞(Th17细胞)、CD4+TCD25+Foxp3+调节性T细胞(Treg细胞)占CD4+T细胞的比例,ELISA法检测血清中细胞因子IL-17、TGF-β、IL-6的水平。结果:急性白血病患者外周血中Th17细胞占CD4+T细胞的(1.39±0.24)%,高于对照组的(0.26±0.11)%(P<0.05);急性白血病患者外周血Treg细胞占CD4+T细胞的(11.58±2.17)%,高于对照组的(2.47±0.72)%(P<0.05);且Treg细胞与Th17细胞呈正相关(γ=0.37)。急性白血病患者血清中TGF-β、IL-6、IL-17的水平分别为(26.06±2.43)、(14.66±2.47)、(18.63±2.38)pg/ml,高于对照组的(13.41±1.92)、(1.44±0.29)、(10.34±1.71)pg/ml(均P<0.05)。结论:急性白血病患者外周血中Treg、Th17细胞比例升高,且两者呈正相关;急性白血病患者血清中TGF-β、IL-6、IL-17水平升高,可能影响Treg与Th17细胞的平衡。     

Th17细胞及其相关细胞因子在胃癌中的表达及临床意义

摘 要：［目的］ 检测Th17细胞及其相关细胞因子在胃癌中的表达,探讨其与胃癌临床病理因素之间的关系。［方法］ 收集45例胃癌患者和20例健康体检者外周血,分离单个核细胞（PBMC）,采用流式细胞分析技术检测Th17细胞比例;收集30例手术切除胃癌组织标本,包括癌组织和非癌对照组织,采用免疫组化法检测胃癌组织内Th17细胞浸润情况;采用荧光定量RT-PCR法检测Th17细胞相关因子维甲酸相关孤核受体C（RORc）、IL-17、IL-23、IL-21、IL-22、IL-1β、TGF-β、IL-6 mRNA表达。［结果］ 与健康对照组相比,胃癌患者外周血中Th17细胞比例显著增多;胃癌组织内可见大量CD4+/IL-17+细胞浸润,而非癌对照组织内极少见CD4+/IL-17+细胞;与非癌对照组织相比,胃癌组织内RORc、IL-17、IL-21、TGF-β、IL-6 mRNA表达水平明显增高,但两组间IL-22、IL-23和IL-1β mRNA表达无显著差异。胃癌组织内RORc mRNA表达水平与IL-17、TGF-β、IL-6 mRNA表达水平呈正相关;并且,RORc mRNA表达水平与患者TNM分期有相关性。［结论］ Th17细胞促进了胃癌的发生和进展。     

Notch信号通路对急性B淋巴细胞白血病患者CD4+CD25+CD127dim/-调节性T细胞的调控作用

摘 要：［目的］ 观察急性B淋巴细胞白血病（B-cell acute lymphoblastic leukemia,B-ALL）患者外周血Notch信号通路分子表达和CD4+CD25+CD127dim/-调节性T细胞（regulatory T cells,Treg）水平,评估Notch信号通路对B-ALL患者Treg活性的影响。 ［方法］ 入组31例B-ALL患者和20名对照者,分选血浆和外周血单个核细胞（peripheral blood mononuclear cells,PBMC）,纯化CD4+CD25+CD127dim/- Treg。实时定量PCR法检测PBMC中Notch1~4、Hes1、Hes5 mRNA相对表达量,流式细胞术检测CD4+CD25+CD127dim/- Treg比例,酶联免疫吸附实验检测血浆白细胞介素10（interleukin-10,IL-10）和IL-35水平。使用Notch信号通路抑制剂GSI刺激B-ALL患者分选的PBMC,检测细胞增殖、Treg比例、IL-10和IL-35表达变化。使用GSI刺激B-ALL患者纯化的Treg,与自体PBMC以1∶10比例共培养,检测细胞增殖、IL-10、IL-35、干扰素-γ水平变化。两组间比较采用独立样本t检验或配对t检验。［结果］ B-ALL患者PBMC中Notch受体（包括Notch1、Notch2、Notch3、Notch4）和Notch下游信号分子Hes1、Hes5 mRNA相对表达量均较对照者升高（P<0.001）。B-ALL患者CD4+CD25+CD127dim/- Treg比例高于对照者（8.90%±2.41% vs 4.68%±1.01%,P<0.001）。B-ALL患者外周血IL-10和IL-35水平均高于对照者（P<0.05）。GSI刺激后B-ALL患者PBMC增殖水平与无GSI刺激组差异无统计学意义（P=0.689）,但GSI刺激后Treg比例和IL-10、IL-35水平均较无GSI刺激组降低（P<0.05）。使用GSI刺激Treg后与自体PBMC共培养,其抑制PBMC增殖的能力减弱（P<0.001）,IL-10和IL-35水平减少（P<0.05）,但干扰素-γ水平增加（P<0.001）。［结论］ B-ALL患者外周血中Notch受体表达升高可能诱导CD4+CD25+CD127dim/- Treg数量增加和免疫抑制活性增强。     

Klotho 蛋白通过TGF-β1/Foxp3/RORγt 通路抑制Treg 和Th7 细胞介导的宫颈癌细胞的免疫逃逸

目的: 研究抗衰老蛋白Klotho 对荷宫颈癌小鼠体内调节性T细胞（Treg 细胞）和辅助性T细胞17（Th7）细胞介导宫颈癌细胞免疫逃逸的影响及其作用机制。方法: 建立宫颈癌U14 细胞移植瘤小鼠模型,并设Control 组（正常小鼠组）、Model 组（荷宫颈癌小鼠模型组）、Klotho 处理组（荷宫颈癌小鼠经Klotho 蛋白处理组,200 ng/d)。分别在处理7、14 d 时称取各组小鼠体内宫颈癌移植瘤质量,以流式细胞术检测各组小鼠脾淋巴细胞功能、Treg和Th7细胞比例变化,qPCR检测各组小鼠Treg和Th7细胞关键转录因子Foxp3、RORγt的表达情况,ELISA方法检测各组小鼠脾淋巴细胞培养液中IL-17、IL-6、IL-10、TGF-β、IL-23 等因子的含量变化,WB检测各组小鼠脾淋巴细胞中Klotho、TGF-β1、Foxp3、RORγt 蛋白表达的变化。结果: 14 d 时,Klotho 组宫颈癌荷瘤小鼠肿瘤抑瘤率显著高于Model 组[(52.16±8.25)% vs (23.33±6.29)%,P<0.05]。Model 组与Control 组相比,荷瘤小鼠脾淋巴细胞中Treg、Th7 细胞比例均显著升高（均P<0.05）,总T淋巴细胞（CD3+）、辅助/诱导T淋巴细胞（CD3+CD4+）比例及免疫指数（CD3+CD4+/CD3+CD8+）显著下降（均P<0.05）,Foxp3、RORγt 基因mRNA表达显著增加（均P<0.05）,IL-17、IL-6、IL-10、TGF-β、IL-23 等因子分泌显著增加（均P<0.05）,Klotho 蛋白水平明显下降（P<0.05）,TGF-β1、Foxp3、RORγt 蛋白表达均明显升高（均P<0.05）;而Klotho 处理组与Model 组相比,上述指标均出现了相反的变化（均P<0.05）,但与Control 组无显著差异（均P>0.05）。结论: Klotho 蛋白可能通过调控TGF-β1/Foxp3/RORγt信号通路抑制宫颈癌荷瘤小鼠体内Treg和Th7 细胞介导的免疫逃逸,从而发挥抗肿瘤作用。     

胃癌患者外周血Th17和Treg细胞的特异性转录因子与相关细胞因子的检测及临床意义

目的 了解胃癌患者外周血Th17和Treg细胞的转录因子及其相关细胞因子的表达水平,探讨其在胃癌发展进程中的作用.方法 收集57例胃癌患者、31例胃部良性疾病患者和40例健康志愿者的外周血,采用实时荧光定量逆转录聚合酶链反应(QRT-PCR)技术,检测外周血单个核细胞中Th17和Treg细胞的特异性转录因子RORγt和FoxP3 mRNA的表达水平;采用酶联免疫吸附试验(ELISA),检测胃癌患者血浆中IL-17、IL-23、转化生长因子β(TGF-β)和IL-10的浓度.结果 胃癌组FoxP3和RORγt mRNA的表达水平分别为2.349±0.491和0.794±0.304,明显高于健康对照组和良性疾病组(均P<0.05),FoxP3/RORγt比值也高于健康对照组和良性疾病组(P<0.05),而良性疾病组与健康对照组差异无统计学意义(P>0.05).中晚期胃癌组的FoxP3/RORγt比值明显高于早期胃癌组(P<0.05).胃癌组血浆中IL-17、IL-23、TGF-β和IL-10的水平明显高于健康对照组(P<0.05),中晚期胃癌组TGF-β和IL-10水平明显高于早期胃癌组(P<0.05).结论 胃癌患者存在高Th17和Treg细胞水平,且随着病程的进展Treg细胞仍维持强势,FoxP3/RORγt比值明显增高.监测胃癌患者外周血Th17和Treg细胞的特异性转录因子和相关细胞因子水平,有助于对病程的判断.     

JAK-STAT信号通路与骨髓增生异常综合征Th17/Treg细胞免疫失调相关性的研究

目的:研究骨髓增生异常综合征（myelodysplastic syndrome,MDS）患者外周血中Th17与Treg细胞比例及其相关细胞因子白细胞介素-2（IL-2）、白细胞介素-6（IL-6）和转化生长因子-β1（TGF-β1）的表达及意义。方法:采用流式细胞术对健康对照组、初诊MDS患者、初诊急性髓系白血病（acute myeloid leukemia,AML）患者的外周血Th17细胞和Treg细胞进行检测,采用双抗体夹心酶联免疫吸附法(ELISA)分别检测上述三组患者外周血IL-2、IL-6与TGF-β1分泌水平,应用RT-PCR和Western blot实验检测STAT3、STAT5 mRNA与蛋白表达水平。结果:与健康对照组相比,初诊MDS患者和AML患者外周血中Treg细胞比例增高（P＜0.05）,但MDS患者组与AML患者组之间没有明显差异（P＞0.05）,三组受试者之间Th17细胞比例无明显差异（P＞0.05）;与健康对照组相比,初诊MDS患者组IL-2表达明显升高（P＜0.05）,初诊AML患者组TGF-β1和IL-2表达均显著增加（P＜0.05）;而MDS患者组与AML患者组相比,TGF-β1、IL-6、IL-2表达水平均无明显差异（P＞0.05）;与健康对照组相比,初诊AML患者组、MDS患者组外周血中STAT5 mRNA与蛋白表达均明显增加（P＜0.05）,但两者之间无差异（P＞0.05）。结论:IL-2/STAT5信号通路可能调节初始Th细胞向Treg细胞转化的关键点,也支持Treg细胞数量增高与MDS的病情进展及其向白血病转化可能有关的论断。     

复方中药联合TP方案治疗晚期非小细胞肺癌的临床疗效观察

目的 探讨复方中药艾迪注射液联合多西他赛+奈达铂(TP)化疗方案治疗晚期非小细胞肺癌(NSCLC)的临床疗效及对Treg/Th17的影响.方法 采用回顾性分析的方法,将95例晚期NSCLC患者按照治疗方法的不同分为对照组47例和观察组48例.对照组患者给予TP化疗方案进行治疗,观察组患者在TP化疗方案的基础上加用艾迪注射液进行治疗.比较两组患者的临床疗效和不良反应发生情况,流式细胞仪检测Treg、Th17细胞比例,液相芯片检测血清IL-10、TGF-β、IL-17、IL-23含量.结果 治疗后,观察组患者的临床获益率为91.7%,高于对照组的76.6%,差异有统计学意义(P=0.044);观察组患者的3~4级中性粒细胞减少、血小板减少的发生率分别为4.2%和2.1%,均低于对照组患者的17.0%和12.8%,差异有统计学意义(P=0.044、0.028);治疗前,两组患者的各指标比较,差异无统计学意义(P﹥0.05);治疗后,两组患者的Treg、Th17细胞比例及相关细胞因子(IL-10、TGF-β、IL-17、IL-23)的含量较治疗前降低,Treg/Th17比例较治疗前升高(P﹤0.05);且观察组患者的各指标均优于对照组患者(P﹤0.05).结论 艾迪注射液联合TP方案治疗晚期NSCLC的疗效肯定,安全性好,且可调节Treg/Th17,值得临床推广.     

转录因子MafB在非小细胞肺癌中的表达及对CD14+单核细胞分泌Ⅰ型干扰素的影响

摘 要：［目的］ 研究非小细胞肺癌（NSCLC）患者中转录因子MafB的表达变化，并评估MafB对NSCLC患者CD14+单核细胞分泌Ⅰ型干扰素和诱导CD4+T细胞分化的影响。［方法］ 入组41例NSCLC患者（28例鳞癌和13例腺癌）和21例健康志愿者。收集外周血和支气管肺泡灌洗液（BALF），分离血浆和外周血单个核细胞（PBMC），纯化CD14+单核细胞和CD4+T细胞。酶联免疫吸附实验检测血浆和BALF干扰素（IFN）-α和IFN-β的水平，反转录实时定量PCR检测外周血和BALF中MafB mRNA相对表达量，Western blot检测MafB蛋白水平。利用MafB siRNA转染CD14+单核细胞，观察抑制MafB对CD14+单核细胞分泌IFN-α和IFN-β的影响，检测干扰素调节因子3（IRF3）磷酸化水平。建立CD14+单核细胞和CD4+T细胞的直接接触和间接接触培养系统，通过ELISA法检测培养上清中IFN-γ、白细胞介素（IL）-4、IL-17和IL-21表达水平评估抑制MafB对CD14+单核细胞调控CD4+T细胞分化的影响。［结果］ 血浆IFN-α和IFN-β水平在健康志愿者和NSCLC患者之间差异无统计学意义，但肿瘤部位BALF中IFN-α（242.5±59.98pg/ml vs 282.5±45.24pg/ml，P=0.013）和IFN-β（12.40±2.81pg/ml vs 34.42±7.83pg/ml，P<0.0001）水平均显著性低于非肿瘤部位。PBMC中MafB mRNA相对表达量和蛋白水平在健康志愿者和NSCLC患者之间差异亦无统计学意义，但肿瘤部位BALF中MafB mRNA和蛋白水平则显著性高于非肿瘤部位（P<0.0001）。MafB siRNA转染可显著性抑制CD14+单核细胞中MafB mRNA和蛋白的表达。MafB siRNA转染可促进CD14+单核细胞IFN-β的分泌（16.09±5.79pg/ml vs 6.73±1.78pg/ml，P<0.0001），增加IRF3磷酸化（P<0.0001），但对IFN-α表达无明显影响（P>0.05）。而抑制NSCLC患者CD14+单核细胞中MafB对CD14+单核细胞和CD4+T细胞共培养系统中IFN-γ、IL-4、IL-17和IL-21的分泌水平并无显著性影响（P>0.05）。［结论］ NSCLC患者肿瘤部位过度表达的MafB可能诱导了Ⅰ型干扰素抑制，但MafB对肿瘤部位CD14+单核细胞的免疫调控功能可能无影响。     

Th17/Treg平衡失调与食管鳞状细胞癌浸润转移的相关性

目的:检测食管鳞状细胞癌患者外周血中辅助性T细胞17(T helper 17,Th17)和调节性T细胞(regulatory T cell,Treg)的表达,探讨食管鳞状细胞癌中Th17/Treg比例的变化与食管鳞状细胞癌转移的关系。方法:收集山东大学齐鲁医院2009年9月至2010年5月食管鳞状细胞癌患者37例,其中男22例、女15例,平均年龄(61±7.85)岁;23例健康志愿者为对照。流式细胞术检测外周血Th17的表达情况,分析Th17占CD3+T细胞的比例,同样流式细胞术检测外周血Treg的表达及其占CD4+T细胞的比例。分析Th17/Treg比例变化与食管鳞状细胞癌病理特征的相关性。结果:食管鳞状细胞癌患者外周血中Th17占CD3+T细胞的比例显著升高[(2.11±0.66)%vs(0.84±0.19)%,P<0.01],Treg占CD4+T细胞的比例也显著升高[(6.21±1.48)%vs(4.43±0.78)%,P<0.01];食管鳞状细胞癌患者转移组与未转移组相比,Th17比例明显升高[(2.42±0.55)%vs(1.61±0.51)%,P<0.01],而Treg比例无明显差异。食管鳞状细胞癌患者外周血中Th17/Treg比例显著高于正常对照组[(0.35±0.13)%vs(0.19±0.04)%,P<0.01];食管鳞状细胞癌患者淋巴结转移组与未转移组相比,Th17/Treg比例显著升高[(0.39±0.13)%vs(0.29±0.10)%,P<0.05]。结论:食管鳞状细胞癌患者外周血中Th17/Treg比例显著升高,可能参与食管鳞状细胞癌的浸润和转移。     

肺癌患者外周血CD+4 CDHi25 CDLo127 调节性T细胞检测的临床意义

 目的　探讨肺癌患者外周血中CD+4 CDHi25 CDLo127 调节性T细胞（Treg）占CD+4 T淋巴细胞比例的变化及其意义。方法　采用流式细胞术检测肺癌患者30例（鳞癌20例、腺癌10例）及健康人（对照组）20名外周血中Treg的变化;ELISA法检测同一标本血清中白细胞介素10（IL-10）,转化生长因子-β（TGF-β）的表达水平。结果　鳞癌、腺癌患者外周血中Treg 占CD+4 T淋巴细胞的比例分别为（6.81±1.52）％、（7.08±1.17）％,两组间比较差异无统计学意义（P＞0.05）,二者均高于对照组的（4.91±1.24）％ ,差异有统计学意义（P＜0.05）;肺癌组血清中IL-10水平为（24.36±4.02）ng／ml,高于健康对照组的（21.53±4.30）ng／ml;肺癌组血清中TGF-β水平为（218.49±35.23）ng／ml,高于健康对照组的（124.31±20.32）ng／ml。结论　肺癌患者外周血中Treg比例明显升高,可能是导致免疫抑制的原因之一,并且可能通过分泌IL-10和TGF-β,发挥免疫抑制的效应。     

Bacteria and cancer--antagonisms and benefits

There is considerable historical and recent evidence concerning the antagonisms between acute bacterial infections or their toxins and cancer and allied diseases. These data provide renewed incentives to undertake clinical programmes with mixed bacterial vaccines in many countries at the present time.     

Religiosity and physical and emotional functioning among African American and White colorectal and lung cancer patients

The literature suggests that religiosity helps cope with illness. The present study examined the role of religiosity in functioning among African Americans and Whites with a cancer diagnosis. Patients were recruited from an existing study and mailed a religiosity survey. Participants (N = 269; 36% African American, 56% women) completed the mail survey, and interview data from the larger cohort was utilized in the analysis. Multivariate analyses indicated that in the overall sample religious behaviors were marginally and positively associated with mental health and negatively with depressive symptoms. Among women, religious behaviors were positively associated with mental health and negatively with depressive symptoms. Religiosity was not a predictor of study outcomes for men. Among African Americans, religious behaviors were positively associated with mental health and vitality. Among Whites, religious behaviors were negatively associated with depressive symptoms. These findings suggest a mixed role of religious involvement in cancer outcomes. The current findings may have applied potential in the areas of emotional functioning and depression.     

Renal irradiation and the pharmacology and toxicity of methotrexate and cisplatinum

We used a rat model to study the effects of renal irradiation on the pharmacology of methotrexate (MTX) and cisplatinum (cis-Pt). Unanesthetized rats were given bilateral kidney irradiation (20 Gy in 9 fractions). At 9 months after irradiation, 3% of the animals had died and survivors showed moderately impaired renal function. At 15 months, 30% of the animals had died and survivors showed severely impaired renal function. Some animals were given i.v. MTX 1 week to 15 months after irradiation. In irradiated rats, the area under the MTX plasma clearance curve equaled that of controls through 6 months, and was significantly above controls from 9 months on. Other animals were given i.p. cis-Pt 1 week to 9 months after irradiation. The acute toxicity of cis-Pt was the same in control and irradiated rats when cis-Pt was given immediately before or after irradiation. Beginning 3 months after irradiation there was a progressive increase in cis-Pt toxicity and a simultaneous decrease in urinary platinum excretion. Irradiated animals that survived cis-Pt treatment showed increased radiation nephritis; the greatest effect occurred when cis-Pt was given 3 months or more after irradiation. MTX and cis-Pt clearance decreased when renal dysfunction was first observed and changes in renal function preceded changes in drug clearance and toxicity.     

Salivary and serum proteomics in head and neck carcinomas: before and after surgery and radiotherapy

Several body fluids have been evaluated as new sources for cancer biomarker discovery. In this context, salivary and serum proteomics seem promising diagnostic and predictive tools for head and neck diseases. In the present study, we performed a proteomic analysis of saliva and serum from patients presenting head and neck squamous cell carcinoma (HNSCC) and compared the results before and after therapy. In saliva of cancer patients, we observed an altered protein profile, including over-expression of PLUNC and zinc-alpha-2-glycoprotein. Both proteins may contribute to control tumor growth and, therefore, represent targets for new analysis. We also detected serotransferrin and a modified transthyretin form with altered levels in serum from patients. Comparing preoperative and postreatment samples, the results showed that the protein profile after treatment reverted to a pattern closer to those observed for controls. These results add information on the role of secreted proteins in the cancer process and emphasize the potential of saliva and serum analysis for diagnosis and monitoring of HNSCC.     

Morphine and tumor growth and metastasis

Morphine is an analgesic widely used to alleviate cancer pain. In addition, the perioperative management of pain in cancer surgery patients most often includes opioids. However, there are reports that these drugs may alter cancer recurrence or metastasis. Several mechanisms have been proposed, such as the modulation of the immune response or cellular pathways that control the survival and migratory behavior of cancer cells. The published literature, however, presents some discrepancies, with reports suggesting that opioids may either promote or prevent the spread of cancer. It is of great importance to determine whether opioids, in particular the most widely used, morphine, may increase the risk of metastasis when used in cancer surgery. This review examines the available data on the effects of morphine which influence cancer metastasis or recurrence, including immunomodulation, tumor cell aggressiveness, and angiogenesis, with special emphasis on recently published clinical and laboratory based studies. We further discuss the parameters that may explain the difference between reports on the effects of morphine on cancer.