促癌变剂对淋巴细胞核仁rRNA基因转录的活化作用——rDNA原位杂交荧光显示法

用原位杂交荧光显示法观察了人淋巴细胞在促癌变剂黄芫花提取物(WCE)和12-0-十四烷巳豆醇-13乙酸酯(TPA)处理后,间期核仁rDNA的定位与数量改变,并与丝裂原植物血细胞凝集素(PHA)的效应作了比较,同时用银染色法观察了核仁。对照组淋巴细胞核仁小,原位杂交的rDNA为少数明亮荧光斑和分散的荧光点。经促癌变剂WCE和TPA处理后,银染色的核仁增大,银染颗粒增多,表明rDNA转录活化。原位杂交证明rDNA信号数目明显扩增,许多荧光小点断续相连形成网织状结构,与PHA刺激核仁转录活化的表现一致。对核仁内所含银染颗粒和rDNA荧光斑点数均值的统计学分析表明,WCE、TPA和PHA各加药组均明显多于对照组。3个加药组之间无明显差别。提示两种促癌变剂皆具有刺激核仁rDNA扩增和转录活化的效应。     

Immunohistological analysis of the lymphoid infiltrate in cutaneous malignant melanomas

Summary The immunological phenotypes of the lymphoid cells in 39 cutaneous malignant melanomas have been investigated by staining cryostat sections with a panel of 20 monoclonal antibodies against lymphoid cells and their subsets. Staining was performed by the alkaline phosphatase: anti-alkaline phosphatase (APAAP) method in which the substrate label (red) is easily distinguishable from melanin. The lymphoid infiltrates had an essentially identical composition in all cases, consisting of T-lymphocytes associated with both Langerhans cells and HLA-DR-positive tissue macrophages. B-lymphocytes and natural killer cells were either absent or only present in low numbers. The ratio between T8 (suppressor/cytotoxic) and T4 (helper/inducer) lymphocytes varied and showed no correlation with melanoma subtype, level of invasion or magnitude of lymphocytic response. Examination for markers associated with T-cell activation and/or with cell proliferation revealed that all lesions contained HLA-DR-positive T-lymphocytes, whereas expression of the transferrin receptor and the interleukin-2 receptor (Tac-antigen) occurred mainly in melanomas with a significant inflammatory infiltrate. These data support the concept that malignant melanomas are capable of evoking autologous T-cell immune reactions.     

Monocytes and lymphocytes as active participants in the pathogenesis of experimental shock

We investigated the role played by monocytes and lymphocytes in the pathogenesis of experimental shock. Splanchnic artery occlusion (SAO) shock was induced in anaesthetized rats by clamping splanchnic arteries for 45 min followed by reperfusion. Sham operated animals were used as controls. SAO shocked rats had a decreased survival time (80±11 min, while sham shocked rats survived more than 4 h), increased serum (248±21 U/ml) and macrophage (145±15 U/ml) levels of TNF-, enhanced myeloperoxidase (MPO) activity in the ileum (3.38±0.2 U×10^–3/g tissue), decreased number of monocytes, lymphocytes and neutrophils and a profound hypotension. In addition we found an increased expression of vascular cell adhesion molecule-1 (VCAM-1) on aortic endothelium and a reduced percentage of VLA-4 positive monocytes and lymphocytes. Inhibition of TNF- synthesis, reversed the increased endothelial expression of VCAM-1, increased the percentage of integrin VLA-4 positive leukocytes and improved monocyte, lymphocyte and neutrophil count. Furthermore a passive immunization with specific antibodies raised against VCAM-1 (2 mg/kg, i.v. 3 h before SAO) increased survival, reduced MPO activity in the ileum (0.034±0.04 U×10^–3/g tissue) and improved mean arterial blood pressure. Our data suggest that monocytes and lymphocytes participate in the pathogenesis of splanchnic ischaemia-reperfusion injury and may amplify the adhesion of neutrophils to peripheral tissues.by I. Ahnfelt Rønne     

Psychoneurodendocrine correlates of lymphocyte subsets during healthy ageing

Ageing has been associated with increased cortisol levels and absolute counts of T lymphocytes with memory phenotype. Although the mechanisms underlying these changes are still unknown, it has been speculated that this could be related to a dysfunction in FAS/CD95 expression in naive or memory cells. In this study, we investigated the role of psychoneuroendocrine variables in regulating CD95 expression on lymphocyte subsets. Forty-six elderly subjects (65-91 years) and 33 young adults (20-40 years) were recruited accordingly the SENIEUR protocol. The psychological status was measured by structured clinical interviews, salivary cortisol was assessed along the day (9, 12 and 22h) and peripheral blood lymphocytes were immunophenotyped. The elderly were more stressed, depressed and anxious than the young subjects. Cortisol levels were increased in the elderly, indicating an activation of the hypothalamic-pituitary-adrenal (HPA) axis. We observed reduced counts of CD45RA+CD95+ cells in the elderly compared to young adults. The elderly subjects also showed a reduced expression of CD3 and CD62L in contrast to increased CD95 expression in CD45RA+ cells. The emotional state was positively correlated with the lymphocyte markers. Our data suggest the healthy ageing is associated with psychoneuroendocrine alterations that may be implicated in the regulation of CD95 expression on peripheral T cells.     

Lectin histochemistry of the lymphoid organs of the chicken

Cellular interactions within the immune system are in part mediated via the carbohydrate-rich coat of the cell membrane, the glycocalyx, of which the terminal carbohydrate residues are of particular functional importance. Thus, these carbohydrate residues from thymus, bursa of Fabricius, spleen and bone marrow of 2- and 30-day-old chickens were investigated by lectin histochemistry. In the thymus, mannose as well as N-acetyl-glucosamine (glcNAc)-specific lectins labelled macrophages, epithelial reticulum cells and lymphocytes within the cortex. In the bursa of Fabricius, the brush border of the lining epithelium, the macrophages and the endothelium were labelled by mannose-specific lectins. The follicle-associated epithelium was labelled by a broad spectrum of lectins. Epithelial cells that separated the cortex from the medulla and large mononuclear cells in the cortex were only being labelled by N-acetyl-galactosamine (galNAc)-specific and glcNAc-specific lectins, respectively. In the spleen, lymphocytes of the peri-ellipsoid lymphocyte sheaths and macrophages of the red pulp were labelled by lectins of nearly all sugar specificities. In general, glycotopes of these organs were more intensively labelled in the 2-day-old chicken than in the 30-day-old chicken, indicating changes in glycotope expression during post-hatching development. Thus, cells of the avian immune system are as rich and diverse in their lectin binding sites as their mammalian counterparts, indicating that similar carbohydrate lectin interactions between cells and matrices take place in birds as well.     

504名放射工作人员淋巴细胞微核率分析

目的 了解长期接触低剂量电离辐射的放射工作人员外周血淋巴细胞微核率的总体水平,为采取相应的放射防护措施提供依据。方法 以2019年1—12月504名资料完整的在岗放射工作人员为放射组,以岗前职业健康检查拟从事放射工作的105名健康成年人为对照组,2组均采用常规培养法测定周围血淋巴细胞微核率,分析比较检测结果。结果 放射组微核率高于对照组（P < 0.001）;女性放射工作人员微核率高于男性（P < 0.01）;不同年龄组微核率差异有统计学意义（P < 0.05）,其中年龄41～50岁的微核率高于21～30岁（P < 0.05）;不同工龄组微核率差异有统计学意义（P < 0.05）,其中21～30年的微核率高于0～10年（P < 0.05）;不同工种间的微核率差异有统计学意义（P < 0.01）,放射治疗岗位人员的微核率较其他岗位最高;不同级别的单位放射工作人员微核率中,市级医院最高,其中省级医院与企业（P < 0.05）、市级医院之间（P < 0.01）的微核率的差异有统计学意义。结论 放射工作人员的微核率高于非接触人员,女性的微核率高于男性,放射治疗的工作人员的微核率较高,应加强对此类人群的放射防护与日常管理。     

Human leukocyte response to an endurance race

Summary The response of circulating leukocytes (WBC's) with regard to changes in number, proportion of neutrophils versus lymphocytes and changes in lymphocyte function as well as proportions of T and B cells was studied in eleven men who ran a 20-mile race. A marked leukocytosis was noted 10–15 min after the race with the predominant increase being polymorphonuclear leukocytes (P<0.001). A significant rise in mean serum cortisol levels was also noted (P<0.001) which correlated with both the increase in total WBC's (P< 0.001) and granulocytes (P<0.001), but not lymphocytes. The increase in serum cortisol was inversely correlated with miles of prior training (P<0.001). An increase in lymphocytes from 1767±112/mm³ to 2431±202/mm³ was less than that previously described in short-term exercise. As with short-term exercise the most significant increase in lymphocytes was in B lymphocytes bearing surface immunoglobulin (P<0.0025). However, in contrast to short-term exercise lymphocytes maintained good in vitro response to the mitogen phytohemagglutinin.This study demonstrates that endurance racing produces a more marked granulocytosis and less lymphocytosis than short bouts of exercise. It is suggested that the degree of leukocytosis is stress dependent in that it was positively correlated with serum cortisol and inversely correlated with prior training.Supported in part by a joint fellowship from the National and Wisconsin Kidney Foundations     

嗜酸性粒细胞在小鼠体内的抗原呈递过程

目的探讨嗜酸性粒细胞(EOS)在体内能否将抗原呈递给T淋巴细胞,了解EOS在体内呈递抗原的过程和特征.方法以鸡卵清蛋白致敏和雾化吸入刺激BALB/c小鼠以诱发EOS在气道聚集.收集并纯化气道EOS以荧光素标记后注入小鼠气道,应用荧光显微镜观察EOS的移行过程.将接受气管EOS注入小鼠的气管旁淋巴结取出后制备单细胞悬液,以流式细胞仪检测其中T细胞的增殖反应并鉴定增殖T细胞的亚群.结果荧光标记的EOS注入正常鼠气道后8h即可出现于气管旁淋巴结(19.0个/mm2±1.8个/mm2),24h达高峰(59.2个/mm2±7.2个/mm2),并至少可以维持120h(29.6个/mm2±2.8个/mm2).致敏小鼠气管内注入5×105个接触过抗原的EOS1d后气管旁淋巴结增殖的T细胞百分数(6.9%±0.5%)即明显高于基础对照值(3.2%±0.3%,P<0.01),3d后达到峰值(10.8%±0.8%,P<0.01),7d以后下降(6.1%±0.6%,P<0.05).此外,EOS呈递抗原所引起的T细胞增殖反应具抗原特异性,出现增殖反应的T细胞仅限于CD4+细胞.结论气道EOS在体内可成为抗原呈递细胞,从而促使CD4细胞出现显著的增殖反应.     

中药复方962对老年大鼠外周血淋巴细胞DNA损伤的影响

目的：观察不同年龄组（4月和24月龄）大鼠和连续灌胃给予中药复方962 3个月的老年大鼠的外周血淋巴细胞DNA的损伤情况。方法：利用快速、灵敏的单细胞电泳技术结合激光扫描共聚焦显微镜来观察大鼠外周血淋巴细胞的DNA损伤,以彗星样细胞出现率（计数一定量细胞中彗星样细胞所占的比例）、占彗星长度（“彗星”电泳方向上的最大长度）和尾距（尾部DNA占总DNA的百分比与头尾部中心间距的乘积）来评价淋巴细胞DNA的损伤程度。结果：发现老年大鼠外周血淋巴细胞中彗尾样细胞出现率显著高于青年大鼠（P＜0．01）,复方962不仅能明显降低老年大鼠的淋巴细胞的彗尾样细胞出现率,而且能明显降低老年大鼠外周血彗星样细胞的总彗星长度和尾蹑（P＜0．01）。结论：实验表明复方962对老年大鼠淋巴细胞DNA的损伤有明显的保护作用。