大肠癌患者外周血T细胞亚群和NK细胞活性的检测

目的检测大肠癌患者外周血T淋巴细胞亚群及NK细胞活性的表达,探讨大肠癌患者免疫状况与肿瘤分期的关系。方法采用流式细胞分析法检测60例大肠癌患者和30例正常对照者外周血T淋巴细胞亚群及NK细胞活性。结果与正常组比较,大肠癌患者CD4^＋细胞、CD4^＋／CD5^＋比值、NK细胞活性降低,CD8^＋细胞升高,这种变化随临床分期的增高更加显著,且化疗后较化疗前明显,术后发生转移者较未转移者明显;术后上述变化有所改善。结论大肠癌患者细胞免疫功能紊乱,并随病情进展及肿瘤转移而加重。切除肿瘤有助于改善患者细胞免疫功能。     

红景天多糖对老年小鼠免疫功能的影响

目的 探讨中药红景天多糖在免疫功能调节方面的作用机制.方法 应用流式细胞技术、放射免疫和ELISA方法检测老年小鼠免疫功能T淋巴细胞亚群,外周血白介素-2(IL-2)含量及免疫球蛋白G(IgG)的含量.结果 红景天多糖对老年小鼠T淋巴细胞亚群、外周血IL-2含量及抗体形成均有增强作用(P<0.05).结论 红景天多糖能够提高老年小鼠的免疫功能,是一种良好的免疫调节剂.     

毛细胞白血病外周血淋巴细胞亚群和NK细胞活性的初步观察

用一组抗Ｔ、Ｂ细胞分化抗原的单克隆抗体检测５例毛细胞白血病病人外周血单个核细胞的表面标记。其中３例Ｂ淋巴细胞占优势（ＣＤ２０：５８％、５０％、６７％；ＣＤ２２：４８％、４３％、５０％，ＳｍＩｇ：８０％、４７％、６０％），包括１例伴有ＩｇＭ单珠峰的病人；２例Ｂ细胞表面标记阳性率很低（ＣＤ２０：２０％，１７％；ＣＤ２２；１５％；１０％；ＳｍＩｇ：１７％、１８％）。５例病人外周血ＷＴ３＋、ＷＴ４＋细胞及ＷＴ４／ＷＴ８比值明显低于正常对照组（Ｐ＜０．００１），ＷＴ８＋细胞增高与正常组比较差异无显著意义（Ｐ＞０．０５）。４例病人ＮＫ细胞活性降低，脾切除后的ＮＫ细胞活性明显升高。     

IL-2I、L-21诱导人外周血单个核细胞及其抗肿瘤作用

目的探讨IL-2I、L-21对人外周血单个核细胞（PBMC）的诱导增殖和表型改变的影响及其体外抗瘤作用。方法取PBMC细胞,调整浓度为1×10^6个/ml,分为四组I,L-2组加入IL-2 100 IU/mlI,L-21组加入IL-21 100IU/ml,联合组加入IL-2 50 IU/ml和IL-21 50 IU/ml,对照组加入0.1 ml生理盐水,台盼蓝染色法计数各组活细胞,流式细胞仪检测各组PBMC表型;以上述四组为效应细胞（E）,以对数生长期的4种肿瘤细胞（人胃癌细胞系M85,胃癌细胞系BGC823,结肠癌细胞系HCT116,结直肠腺癌细胞系HCT8）为靶细胞（T）,稀释靶细胞5×10^3个/孔,E∶T为1∶1和2∶1,MTT法测定细胞毒性（杀伤率）。结果联合组活细胞计数高于IL-2组I、L-21组、对照组,P均〈0.05;联合组I、L-2组和IL-21组CD3^-/CD56^＋、CD3^＋/CD56^＋水平高于对照组,P〈0.05或0.01;联合组对4种肿瘤细胞的杀伤率均高于其余三组I,L-2组I、L-21组高于对照组,P均〈0.05。结论 IL-2联合IL-21的协同刺激作用,可有效地刺激PBMC的增殖和表型改变,对4种消化道肿瘤细胞株均有较强的杀伤力。     

化疗肺癌患者外周血T淋巴细胞亚群水平分析

目的了解肺癌病人化学治疗后体内T淋巴细胞亚群的变化情况,为临床化疗和免疫调节提供依据和指导。方法用流式细胞术检测化疗后肺癌病人体内各T淋巴细胞亚群的百分比和绝对计数值,并和健康人群进行比较,进行综合分析。结果化疗的肺癌病人CD3^＋T淋巴细胞百分比为（64．43±12．48）％,CD4^＋T淋巴细胞百分比为（35．75±8．69）％,与对照组比较差异均有统计学意义（t=3．838,P=0．000;t=6．188,P=0．000）;CD8^＋T淋巴细胞百分比为（24．23±11．74）%,与对照组比较无统计学差异（P〉0．05）。但CD8^＋T淋巴细胞绝对计数与对照组比较,差异有统计学意义（t=8．473,P〈0．05）。结论化疗的肺癌病人免疫功能显著受损,而且以细胞免疫功能受损为主,临床上进行免疫调节时应考虑这一因素。     

低氧对人外周血单个核细胞迁移活性的影响及其临床意义

目的探讨低氧对机体外周血单个核细胞（PBMC）迁移的影响及其临床意义。方法分离外周血单个核细胞,采用Transwell小室实验检测常氧和低氧条件下PBMC迁移能力的变化,流式细胞术（FCM）检测常氧和低氧条件下T、B淋巴细胞,NK细胞及单核细胞迁移的变化。结果低氧环境下PBMC的迁移率降低31％。对其细胞亚群的分析结果显示,NK细胞的迁移率降低31．7％,单核细胞的迁移率降低63．1％,T、B淋巴细胞的迁移率无显著变化。结论低氧能显著影响单核细胞及NK细胞的迁移功能;检测低氧条件下PBMC的迁移能力,可为以免疫细胞迁移功能为靶点的炎症等疾病提供新的治疗对策。     

仿刺参糖胺聚糖对健康人和肺癌患者外周血单个核细胞体外增殖活性的影响

目的探讨不同浓度的仿刺参糖胺聚糖(HGAG)对健康人和肺癌患者外周血单个核细胞(PBMC)体外增殖活性的影响。方法采用Ficoll密度梯度离心法分别从健康人和肺癌患者外周血分离PBMC,在体外与0、0.1、1、10、50、100μg/m L的HGAG共同培养,CCK-8法检测PBMC增殖的光密度(OD)值。结果 HGAG在1~50μg/m L浓度范围内健康人PBMC的OD值均增高,与0μg/m L相比P均<0.05,浓度为10μg/m L时OD值最高(P均<0.05),0.1、100μg/m L与0μg/m L相比差异无统计学意义;HGAG在10~100μg/m L浓度范围内肺癌患者PBMC的OD值均增高,与0μg/m L相比P均<0.05,且HGAG浓度越高其OD值越高(P均<0.05)。结论HGAG在一定浓度范围内均能够促进健康人和肺癌患者PBMC的增殖,从而有助于提高机体的免疫功能,发挥抗肿瘤作用。     

茶多糖对NOD小鼠1型糖尿病的预防作用

目的探讨茶多糖（TPS）对非肥胖糖尿病（NOD）小鼠1型糖尿病（DM）的预防作用。方法比较TPS预免疫组和生理盐水（NS）对照组NOD小鼠1型DM的发病率、血清C肽和谷氨酸脱羧酶抗体水平、胰岛组织病理学和免疫组化、脾脏T细胞亚群比例。结果TPS预免疫组与NS组比较，DM发病率显著降低，血清C肽水平显著增高，胰岛炎症程度减轻，CD8T细胞亚群比例显著增高，CD4／CD8比例显著降低。结论早期应用茶多糖预免疫可以预防或延缓NOD小鼠1型糖尿病的发生。     

慢性HBV感染者外周血T淋巴细胞亚群和NK细胞活性变化

探讨慢性HBV感染者外周血T淋巴细胞亚群及NK细胞活性变化情况。应用流式细胞法检测所有研T淋巴细胞亚群和NK细胞。慢性乙肝、肝炎肝硬化和慢性重型乙肝组患者CD3 、CD4 百分率及CD4 ／CD8 比值与正常组比较均有所下降,且慢性重型乙肝组患者CD4 百分率和CD4 ／CD8 比值与正常组比较差异有显著性。各临床类型慢性HBV感染者NK细胞百分率均降低,与正常对照组比较有统计学意义。慢性HBV感染者细胞免疫功能低下。检测T淋巴细胞亚群及NK细胞活性变化对判断病变程度、指导临床治疗具有一定参考价值。     

Protease-induced immunoregulatory activity of platelet factor 4

Intravenous injection of human or mouse serum or platelet material secreted from appropriately stimulated platelets ("releasate") together with antigen alleviates the immunosuppression in SJL/J mice induced by injection of irradiated lymphoma cells or in (CB6)F1 mice induced by injection of concanavalin A. We now report that injection of releasate from 10(6) human platelets restores plaque-forming cells to the unsuppressed number; greater amounts increase responses further. Immunoregulatory activity is released from platelets exposed to thrombin in parallel with other alpha-granule components. Heparin-agarose absorbs activity. Purified platelet factor 4 (PF4) has activity; beta-thromboglobulin and platelet-derived growth factor have little or none. Activity in serum is neutralized by goat anti-human PF4. An enzymatic step is necessary for production of immunoregulatory activity. Releasates boiled immediately after platelet aggregation with 250 nM A23187 or those produced by adding A23187 in the presence of 100 microM serine protease inhibitor (p-amidinophenyl)methanesulfonyl fluoride (APMSF) are ineffective, whereas releasates boiled or mixed with APMSF after incubation for 60 min are active. Activity is generated by incubating a mixture of heparin-absorbed releasate (as enzyme source) and heparin-agarose eluate of releasate made in the presence of APMSF (as substrate source). The enzymatic step does not alter the heparin-neutralizing activity of PF4. Apparently a secreted platelet protease converts PF4 to a form with immunoregulatory activity.     

Effects of propantheline on the motor activity of the normal human esophagus

The aim of this study was to examine the action of orally administrated propantheline bromide, an anticholinergic agent, on esophageal motor function. To evaluate these effects a double blind randomized study was carried out in ten normal volunteers. An optimal effective dose for each subject was determined according to the Sun and Shay method. Esophageal motor activity following dry and wet swallow was markedly altered by propantheline. This drug: a) dramatically lowered the peristaltic wave amplitude in the smooth muscle part of the esophagus (P less than 0.001); b) decreased the wave velocity in the proximal part of the smooth muscle (P less than 0.05), and c) increased the frequency of both the non peristaltic and repetitive waves (P less than 0.001). On the other hand propantheline weakly but significantly diminished the lower esophageal sphincter resting pressure (P less than 0.05). These results suggest that: a) the peristaltic function of the human esophageal smooth muscle is greatly dependent on muscarinic neurotransmission, and b) in patients with gastroesophageal reflux, orally as well as parenterally administrated anticholinergic agents are contraindicated.     

Lyophilized whole human melanoma cells stimulate human PBMC proliferation and enhance suppressive action of PBMC toward survival of the same malignant cell line in vitro

The goal of this work was to determine: a) do lyophilized human melanoma BG or Fem-X cells affect the proliferative capacity of normal human peripheral blood mononuclear cells (PBMC) and b) does the PBMC six-days preincubation in nutrient medium with FBS with, or without lyophilized human melanoma BG or Fem-x cells, affect their suppressive action on the survival of the same malignant cell line in vitro. In the aim to avoid any stimulating effect of FBS, other group of experiments were done in nutrient medium with human AB serum in order to determine: c) does the PBMC six-day-preincubation with lyophilized human melanoma BG or Fem-x cells affect their antiproliferative action on the corresponding malignant cell line in vitro and d) does the PBMC six-day preincubation with lyophilized normal PBMC, obtained from healthy volunteer (as a source of allogenous, but not of tumor antigens), affect their suppressive action on the survival of both melanoma BG and Fern-x cell lines in vitro. Results obtained in the presence of FBS in nutrient medium, showed that lyophilized BG cells induced a proliferation of the healthy PBMC, depending on the number of stimulating lyophilized cells. Lyophilized Fem-x cells induced healthy PBMC proliferation in lesser degree than lyophilized BG cells. This stimulation was almost constant, not dependent on the number of stimulating lyophilized Fem-x cells. Six-day stimulation in vitro by both lyophilized melanoma cells enhanced the suppressive action of PBMC on the survival of the corresponding malignant cell line. Experiments done in nutrient medium with normal human AB serum showed that six-day stimulation with lyophilized melanoma cells enhanced, again, the suppressive action of PBMC on the survival of the corresponding malignant cell line. Contrary, six day preincubation of normal PBMC with the lyophilized healthy PBMC (obtained from other healthy person) inhibited their suppressive action on the survival of both malignant cell lines in vitro.     

树舌多糖对胃癌MGC-803细胞蛋白激酶A活性的影响

目的 研究树舌多糖(GAPS)对胃癌MGC-803细胞增殖抑制与细胞蛋白激酶A(PKA)活性的关系,探讨GAPS抗肿瘤作用机制.方法 倒置显微镜观察细胞形态;PepTag非放射法检测PKA活性.结果 GAPS组细胞形状不规则,细胞间隙增大折光性差,生长状态差;GAPS组PKA活性的上调,与细胞对照组比较有差异(P＜0.05).结论 GAPS能抑制胃癌MGC-803细胞的增殖,上调PKA活性.     

Effects of nutrient liquids on human gastroduodenal motor activity

The effects of intragastric infusion of 10% Intralipid and 10% dextrose on the intraluminal pressures in the antrum, pylorus and duodenal bulb have been examined. Ten studies with each infusate have been performed in 10 normal subjects and the results compared with those obtained previously in 22 studies during intragastric infusion of isotonic saline. During saline infusion, contractile activity varied. In six studies fasting motor activity persisted; in the remainder, variable activity, without recognisable pattern was recorded. With saline, the gastroduodenal region usually functioned as a unit and the pylorus was the least autonomous part. Neither a sustained rise of basal pressure nor rhythmic, independent contractions were recorded from the pylorus. The contractile activity of the gastroduodenal region with Intralipid and dextrose was more uniform than with saline. Fasting motor activity was always abolished. The gastroduodenal region ceased to contract as a unit and the pylorus acquired autonomous activity. Rhythmic, independent contractions of the pylorus were recorded in nine of 10 studies during Intralipid infusion and six of 10 studies with dextrose. In addition, a sustained rise in pyloric basal pressure was recorded in eight of 10 studies with Intralipid and three of 10 studies with dextrose. Pyloric motility indices were significantly greater with fat than with dextrose. The observed differences in gastroduodenal motility are consistent with a role for the pylorus in the control of emptying of liquid from the stomach.     

Spectrum of immunoregulatory functions and properties of human alveolar macrophages

In health, pulmonary alveoli are maintained free of inflammatory responses to inhaled foreign antigens. The specific role of alveolar macrophages (AM) in modulating the local cellular immune response to antigens is controversial. Immunoregulatory function and properties of AM and blood monocytes (MN) were compared. The AM were obtained by bronchoalveolar lavage of healthy volunteers, MN by adherence of peripheral blood mononuclear cells to plastic. These accessory cells were added in increasing ratios to a responder population rendered rigorously accessory cell-dependent by nylon wool adherence and depletion of cells bearing the surface Class II MHC determinant, HLA-DR. At low ratios of mononuclear phagocytes to lymphocytes (less than or equal to 1:10), MN and AM supported significant and comparable blastogenic responses to tetanus toxoid (3H-thymidine incorporation at a 1:10 ratio was 9,697 +/- 2,508 for MN and 8,969 +/- 1,454 for AM, mean cpm +/- SE, n = 9, p = NS) and other antigens. Interleukin-1 (IL-1) activity in supernatants of MN stimulated with lipopolysaccharide (LPS), 10 micrograms/ml, was 115 +/- 28 versus 67 +/- 21 U/ml in supernatants of AM (n = 9, p greater than 0.2). At suboptimal concentrations of LPS, however, MN expressed more IL-1 activity than did AM. The specific mean fluorescence intensity of surface expression of HLA-DR determinants as assessed by flow cytometry was similar for MN and AM. At the higher ratio of 1:2, MN supported 32% increased responses to tetanus toxoid compared with that at 1:5 (p less than 0.05). In contrast, AM at a ratio of 1:2 suppressed lymphocyte response by 69% (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Biological and immunological characterization of a human liver immunoregulatory protein

The liver immunoregulatory protein (LIP) was originally characterized as human liver-derived soluble factor which inhibited the alloantigen and phytohemagglutinin-induced proliferation of human lymphocytes (1). Soluble extracts prepared under the same experimental conditions from kidney, spleen, heart, lymph nodes, and erythrocytes did not exert any inhibitory activity (2). The purpose of this study was to characterize the immunobiological properties of LIP. In the primary one-way mixed lymphocyte culture, LIP depressed the generation of suppressor T cells which inhibited the lymphocyte proliferation induced by phytohemagglutinin or alloantigens. In addition, LIP suppressed in primary mixed lymphocyte culture the induction of cytotoxic T cells and memory cells as determined by cell-mediated lympholysis and secondary mixed lymphocyte culture, respectively. In the presence of LIP, the concanavalin A-mediated induction of suppressor T cells, the pokeweed mitogen-induced IgG synthesis in vitro and the cytolytic activity of K cells reacting in the antibody-dependent cell-mediated cytotoxicity were also inhibited. Cytotoxic effects could be excluded since the viability of human lymphoblastoid cells, hepatocytes, and allogeneically stimulated lymphocytes was not affected by LIP. LIP was shown to be different from other liver-derived substances like acute phase proteins, immunoregulatory alpha-globulins, C-reactive protein, lipoproteins, and F antigen. Furthermore, LIP is not identical to other serum components like the immunoregulatory rosette inhibition factor and the serum inhibitory factor (3). However, the characteristics described herein strongly indicate that LIP is very similar to the liver extract described by Chisari (4) and the liver-derived inhibitory protein (LIP) described by Grol and Schumacher (5).(ABSTRACT TRUNCATED AT 250 WORDS)     

树舌多糖抑制胃癌MGC-803细胞增殖及对酪氨酸激酶活性的影响

目的探讨树舌多糖对人胃癌MGC-803细胞增殖、细胞周期分布及其对酪氨酸激酶表皮生长因子受体(EGFR)、血小板衍生生长因子受体β(PDGFRβ)表达的影响。方法 MTT法检测树舌多糖对胃癌细胞MGC-803增殖抑制;流式细胞仪检测树舌多糖作用后MGC-803细胞的周期分布、EGFR及PDGFRβ表达变化;荧光显微镜下观察细胞EGFR、PDGFRβ表达情况。结果树舌多糖对胃癌MGC-803细胞增殖具有抑制作用,并呈时间及剂量依赖性。2mg/ml树舌多糖作用MGC-803细胞2h后,细胞周期阻滞于G0/G1期,S期和G2/M期细胞减少,与对照组细胞比较有差异(P0.05)。树舌多糖下调MGC-803细胞EGFR、PDGFRβ的表达,与对照组细胞比较有差异(P0.05);荧光显微镜和流式细胞仪检测结果一致。结论树舌多糖抑制胃癌MGC-803细胞增殖,下调酪氨酸激酶EGFR、PDGFRβ表达和阻止胃癌细胞由G1期进入S期,延缓细胞周期进程。