Suppressive effects of antigens on the activity of specific activated lymphocytes：A test to define the specificity of activated lymphocytes

Objective：With the regular mixed lymphocytes culture （MLC） to detect the allograft rejection, the reactivity of the activated lymphocytes （primed lymphocytes） of a recipient shows sometimes increase and sometimes decrease against the antigens from the donor, which is inconsistent with the clinical results. In order to establish a convenient method for testing the specificity of the activated lymphocytes in vitro, so as to know the rejection occurred or not by testing the existence of the specific activated lymphocytes against donor＇s HLA antigens in the recipient＇s peripheral blood. Methods： Anti-IL-2 neutralizing monoclonal antibody （anti-IL-2 N-mAb） and immunosuppressors were introduced in this test system in the presence of specific stimulators and activated lymphocytes. Results ： When the activated lymphocytes were chosen from the one-way MLC 4 d to undergo re-stimulation by specific stimulators, the activity of activated lymphocytes in the treatment group was suppressed significantly compared with that in the control group. The result of this test method is consistent with the biopsy in the clinical diagnosis of rejection. Conclusion：h suggests that the activated lymphocytes can be inactivated by specific antigens in certain conditions. This can be a useful tool to define the specificity of the activated lymphocytes.     

人脐血来源树突状细胞体外扩增及诱导特异性抗宫颈癌免疫应答

目的:研究脐血来源树突状细胞(DC)体外扩增及诱导特异性抗宫颈癌细胞的免疫效应。方法:自人脐血分离单核细胞诱导DC,制备宫颈癌细胞系CaSki冻融物作为抗原负载DC。ELISA法检测成熟DC上清中IL-12的含量,混合淋巴细胞反应(MLR)测定DC体外刺激T细胞增殖的能力,MTT法检测抗原负载DC激活的CTL对宫颈癌细胞CaSki和HeLa的杀伤作用。结果:与未经抗原负载的DC相比,抗原负载的DC刺激同种异体T细胞增殖和IL-12分泌的能力均有提高(P<0·05),激活的CTL对CaSki细胞有特异性杀伤,而此CTL对HeLa细胞无特异性杀伤。结论:宫颈癌细胞CaSki冻融抗原负载DC激活的CTL可诱导出特异性杀伤宫颈癌细胞的免疫效应。     

新穿心莲内酯对小鼠巨噬细胞呼吸爆发及淋巴细胞增殖的影响

目的: 观察新穿心莲内酯对小鼠RAW264. 7巨噬细胞呼吸爆发和BALB/C小鼠淋巴细胞增殖的影响,探讨其抗炎免疫活性。方法:分别用化学发光法和AlamarBlue染色法考察新穿心莲内酯对佛波豆蔻酸乙酯刺激的小鼠巨噬细胞呼吸爆发及植物凝集素刺激的BALB/C小鼠脾淋巴细胞增殖的影响。结果:新穿心莲内酯在 75 ~150μmol·L-1浓度范围内对刺激的巨噬细胞呼吸爆发及淋巴细胞增殖有抑制作用,而在 1. 5~7. 5μmol·L-1浓度范围内可协同刺激剂引起的呼吸爆发。结论:新穿心莲内酯在高浓度时具有抗炎活性,而低浓度时具有免疫刺激活性,提示其可能具有双向调节功能。     

Use of the tail moment of the lymphocytes to evaluate DNA damage in human biomonitoring studies.

The Comet assay has gained increasing popularity for use in human biomonitoring or epidemiologic studies; however, one of the shortcomings of the Comet assay is a lack of agreement on a single appropriate Comet parameter that is capable of adequately describing observed DNA damages. Among the tail parameters of Comet features, the most frequently used are the tail moments (both the Olive tail moment and the extent tail moment), the tail DNA, and the tail length. Some studies comparing Comet parameters have been found in cell toxicity research, but there are few comparative studies that use human biomonitoring or epidemiologic data. In this study, we evaluate those four tail parameters in both high and low DNA damaged cells with the use of epidemiologic data. To do this, a new graphical approach, the so-called quantile dispersion graphs (QDGs) are used. In a comparison of an exposed group and a control group, either the tail moment or tail DNA is preferable to the tail length. With respect to providing smaller variability in quantiles for the amount of DNA damage, however, the tail moment is the preferred parameter for both groups. Moreover, the tail moment provides the most stable estimates for DNA damage because it has a larger degree of uniformity in quantile dispersions. To study high degrees of damage from toxic exposure using B cells or G cells, however, the tail DNA showed more significant discrepancies than the other parameters, in terms of both the mean differences and the graphical differences between the two groups. In view of this result, it is suggested that both the tail moment and the tail DNA be presented as tail parameters in human biomonitoring studies.     

III期结直肠癌中NLRC3与预后和肿瘤免疫的关系

目的：探讨核苷酸寡聚化结构域样受体家族半胱天冬酶募集结构域蛋白3(nucleotide binding oligomerization domain-like receptor family caspase recruitment domain containing 3,NLRC3)与III期结直肠癌的预后及肿瘤免疫相关指标 的关系。方法：回顾性收集中南大学湘雅医院2012年至2013年122例经手术根治切除的III期结直肠癌患者的相关资 料。利用免疫组织化学法分析NLRC3与CD8+ T细胞的表达情况,利用患者的术前临床资料计算中性粒细胞/淋巴细 胞比值(neutrophil to lymphocyte ratio,NLR),并检测其微卫星稳定性。采用χ2检验分析NLRC3与临床病理因素之间的 关系,采用COX回归模型分析III期结直肠癌的独立预后因素。结果：在III期结直肠癌中,肿瘤组织CD8+ T细胞浸润 分期(χ2=27.79,P<0.01)、NLR值(χ2=6.35,P<0.05)、淋巴结转移分期(χ2=10.12,P<0.01)以及微卫星稳定性(χ2=6.05, P<0.05)与NLRC3的表达有关。NLRC3(OR=0.066,95% CI：0.020~0.218)、血管癌栓(OR=3.119,95% CI：1.547~6.286) 及NLR(OR=5.103,95% CI：2.465~10.563)对III期结直肠癌的5年总生存期(overall survival,OS)有影响(均P<0.05);另 外,NLRC3(OR=0.144,95% CI：0.055-0.377)、血管癌栓(OR=3.589,95% CI：1.859~6.932)及NLR(OR=2.939,95% CI： 1.509~5.723)对III期结直肠癌的无病生存期(disease free survival,DFS)同样有影响(均P<0.05)。结论：NLRC3,血管癌栓 和NLR是III期结直肠癌的独立预后因素。NLRC3通过抑制系统性炎症、促进局部抗肿瘤免疫而使III期结直肠癌患者 具有良好的预后。     

Prognostic impact of the preoperatory neutrophil/lymphocyte index on early surgical complications of patients with colorectal cancer

Colorectal cancer (CRC) is found among those with greatest frequency and exponential increase worldwide, with high mortality rates, which are observed as increasing due to the postsurgical complications that come to present. Systemic inflammation participates in the development and progression of cancer; therefore, inflammatory and/or immunological response markers such as the neutrophil/lymphocyte index (NLI) can aid us in predicting the poor results of our interventions. The purpose of our study was to determine the impact of an NLI of ≥2.6 as a predictor of early postsurgical complications. By means of a prospective cohort, we analyzed 158 patients with CRC who were submitted to elective surgery with a later 30-day follow-up. We found that the preoperatory NLI of ≥2.6 obtained an odds ratio (OR) = 2.24 (95% confidence interval [CI], 1.15-4.36) as a prognostic factor of early postsurgical complications according to the Clavien-Dindo classification scale, which represents a low prognostic impact due to its predictive yield with low accuracy, which is the opposite of what other reports have previously published. The use of chemotherapy before the surgical procedure was also determined to be a risk factor for post-surgical complications.     

茶多酚有效成分对淋巴细胞增殖的抑制和抗氧化作用

研究表明,茶多酚具有很强的消除有害自由基、抗衰老等多种药理作用[1],茶多酚临床治疗白癜风具有较明显的效果,但其各类组分在临床治疗白癜风所发挥的作用还未见报道。1材料与方法1.1动物C57BL/6(B6)小鼠,体质量(20±2)g,由浙江中医药大学实验动物中心提供。1.2主要药品与试剂H2O2、EGCG均购自美国Sigma公     

六味地黄汤相关苷类单体对小鼠脾淋巴细胞增殖作用的研究

目的：研究六味地黄汤中芍药苷、薯蓣皂苷元、马钱苷、莫诺苷、桃叶珊瑚苷对小鼠T、B淋巴细胞增殖的影响。方法：制备脾淋巴细胞,MTT比色法检测不同浓度5种苷类单体单独作用及其与丝裂原（ConA、LPS）共同作用时对小鼠T、B淋巴细胞增殖作用的影响。结果：10-5M芍药苷能显著促进淋巴细胞增殖,对LPS诱导的B淋巴细胞增殖有明显的协同作用,10-7～10-6M芍药苷对ConA诱导的T淋巴细胞增殖有明显协同作用,与对照组比较有显著性差异（P〈0.01,P〈0.05）;薯蓣皂苷元有一定促进淋巴细胞增殖作用,但与对照组比较无显著性差异;10-7～10-5M马钱苷能显著促进淋巴细胞增殖,10-7M马钱苷对LPS诱导的B淋巴细胞增殖有显著协同作用,随剂量增加,增殖作用反而降低;10-7～10-5M莫诺苷能显著促进淋巴细胞增殖,对LPS诱导的B淋巴细胞增殖有明显的协同作用,对ConA诱导的T淋巴细胞增殖有明显协同作用,与对照组比较有显著性差异（P〈0.01）。10-7～10-5M桃叶珊瑚苷基本无促进淋巴细胞增殖作用1,0-5M桃叶珊瑚苷对LPS诱导的B淋巴细胞增殖有明显的协同作用,10-7～10-5M桃叶珊瑚苷对ConA诱导的T淋巴细胞增殖有明显协同作用,与对照组比较有显著性差异（P〈0.01,P〈0.05）。结论：六味地黄汤中5种苷类单体对T、B淋巴细胞均有不同程度增殖作用,莫诺苷作用最明显,马钱苷低剂量促进增殖,高剂量反而抑制。     

淀粉样β蛋白1-40对爪蟾卵母细胞表达的淋巴细胞神经递质受体功能的影响

观察淀粉样β蛋白_1-40(Aβ_1-40)对爪蟾卵母细胞表达的淋巴细胞神经递质受体(NR)功能的影响,探讨淋巴细胞NR功能检测在阿尔茨海默痴呆(AD)病理生理学上的意义. 将提取的人外周血淋巴细胞mRNA(50 ng)显微注射到成熟的非洲爪蟾卵母细胞,在(19.0±1.0)℃条件下孵育24 h后采用双电极电压钳位记录-60 mV维持电位时的受体激活电流. Aβ_1-40于记录前12 h加入孵育液. 结果发现注射人外周血淋巴细胞mRNA的爪蟾卵母细胞能表达出乙酰胆碱, 谷氨酸, 多巴胺, 5-羟色胺和γ-氨基丁酸受体. 这些表达的受体激活电流的共性是由氯离子载流的内向电流,其平衡电位接近于-22 mV. Aβ_1-40 60 nmol·L^-1对卵母细胞表达的NR功能有抑制效应,其NR峰电流降低分别为乙酰胆碱23%, 谷氨酸27%, 多巴胺25%,维生素E有拮抗Aβ_1-40效应的作用。外周血淋巴细胞NR功能检测作为AD外周评价指标值得更深入的研究.     

三七总皂苷对类风湿关节炎患者环瓜氨酸肽特异性T细胞增殖的影响

目的观察三七总皂苷(PNS)对类风湿关节炎(RA)患者环瓜氨酸肽抗原特异性T细胞(CCP/AST)体外增殖的影响。方法体外分离培养24例RA患者外周血淋巴细胞,分为以下5组,空白对照组:不加任何干预药物;CCP组:仅加CCP干预(CCP终浓度为20μg/mL);甲氨蝶呤(MTX)对照组:同时加CCP与甲氨蝶呤干预(CCP与甲氨蝶呤终浓度均为20μg/mL);PNS低剂量组:同时加CCP与PNS干预(CCP与PNS终浓度分别为20、10μg/mL);PNS高剂量组:同时加CCP与PNS干预(CCP与PNS终浓度均为20μg/mL)。体外培养72h后,用CCK-8法测定T细胞增殖水平。结果 CCP组患者外周血淋巴细胞增殖(0.70±0.08)与空白对照组(0.49±0.06)相比明显升高,差异有统计学意义(P<0.01);PNS低剂量组和高剂量组与CCP组相比,淋巴细胞增殖均可被明显抑制(0.57±0.06、0.47±0.06与0.70±0.08),差异均有统计学意义(P<0.01);且PNS高剂量组与PNS低剂量组相比,差异有统计学意义(P<0.01)。结论 PNS对CCP/AST增殖的抑制作用可能在RA的治疗中发挥重要作用,其抑制程度可能与PNS剂量有一定的相关性。