胃癌组织端粒酶与外周血T淋巴细胞的关系研究

目的探讨胃癌患者外周血T淋巴细胞与胃癌组织端粒酶的关系。方法采用TRAP-ELISA法检测38例胃癌患者、33例正常对照胃黏膜组织端粒酶的活性,采用流式细胞仪(FCM)检测38例胃癌患者、40例健康对照者外周血的T淋巴细胞亚群。结果胃癌组织中端粒酶阳性率78.9%,而正常胃黏膜组织为9.0%,胃癌组端粒酶活性显著高于对照组(P<0.01);胃癌组外周血淋巴细胞亚群CD4+及CD4+/CD8+比值都低于对照组(P<0.01);TNM分期Ⅲ/Ⅳ期患者显著低于Ⅰ/Ⅱ期患者及正常人(P<0.01);端粒酶活性与外周血淋巴细胞两者无相关性(r=-0.136,P<0.05)。结论端粒酶活性在胃癌患者中高度表达;胃癌患者CD4+及CD4+/CD8+降低,表明T细胞介导的细胞免疫功能降低和平衡失调,但端粒酶与T淋巴细胞亚群的改变无相关性。

Analysis of the T-Lymphocytes and Their Subpopu-lations of the Peripheral Blood in Patients with Urticaria

Objective: To study the function of cellular immunity of patients with urticaria. Methods:T-lymphocytes subpopulations of the peripheral Mood ire 60 patients with urticaria and 40 healthy controls were examined by flaw cytometry. Results: The number of CD3+ and CD4+ cells in the urticaria, group were significantly lower than tliose in the control group (P<0. 01) , especially in patients with acute urticaria. Conclusion: There was immunologic dysfunction of T lymphocyte in the patients with urticaria, and not only humoral immunity takes part but also cellular immunity plays a certain role in the pathogenesis of urticaria.

T-regulatory lymphocytes in peripheral blood of gastric and colorectal cancer patients

AIM: To assess the absolute number of T-regulatory cells (Tregs; CD4+CD25+Foxp3+) in the peripheral blood of gastric and colorectal cancer patients. METHODS: We enrolled 70 cancer patients (33 gastric cancer, 37 colorectal cancer) and 17 healthy volunteers. The CD3+CD4+ lymphocytes and CD4+CD25+Foxp3+ Tregs in the peripheral blood were analyzed with flow cytometry. The absolute numbers of Tregs were calculated based on the CD4+CD25+Foxp3+ cells percent-age of CD3+CD4+ cells and the absolute numbers of CD3+CD4+ cells per microliter. RESULTS: The mean number of CD4+CD25+Foxp3+ cells per microliter in colorectal cancer patients was 15.7 (SD: 21.8), for gastric cancer patients 12.2 (SD: 14.3), and for controls 17.5 (SD: 11.4). The absolute number of Tregs was significantly lower in gastric cancer patients than in controls (P = 0.026). There was no statistically significant difference for gastric vs colorectal cancer or colorectal cancer vs controls. The absolute number of Tregs was also significantly depressed in N+ vs Ncancer patients [22.0 (27.7) vs 10.1 (9.0), P = 0.013], and in the subgroup of gastric cancer patients [30.3 (27.6) vs 9.6 (8.0), P = 0.003]. No statistical difference was observed in the proportion of Tregs in the CD4+ population between the groups. CONCLUSION: The absolute number of Tregs in peripheral blood of gastric cancer but not colorectal cancer patients was significantly decreased in comparison with that in healthy controls.

Flow cytometric characterizations of leukocyte subpopulations in the peripheral blood of northern pig-tailed macaques （Macaca leonina）

Pig-tailed macaques(Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. Northern pig-tailed macaques(M. leonina) are distributed in China and other surrounding Southeast Asia countries. Although northern pig-tailed macaques have been bred on a large scale as experimental animals since 2012, the reference value of normal levels of leukocytes is not available. To obtain such information, 62 blood samples from male and female healthy northern pig-tailed macaques at different ages were collected. The normal range of major leukocyte subpopulations, such as T lymphocytes, B lymphocytes, natural killer(NK) cells, monocytes, and the expression levels of activation or differentiation related molecules(CD38, HLA-DR, CCR5, CD21, IgD, CD80 and CD86) on lymphocytes were analyzed by flow cytometry. The counts of B cells decreased with age, but those of CD8+ T cells and NK cells and the frequency of CD38+HLA-DR+CD4+ T cells were positively correlated with age. The counts of leukocyte subpopulations were higher in males than those in females except for CD4+ T cells. Males also showed higher expression levels of Ig D and CD21 within B cells. This study provides basic data about the leukocyte subpopulations of northern pig-tailed macaques and compares this species with commonly used Chinese rhesus macaques(M. mulatta), which is meaningful for the biomedical application of northern pig-tailed macaques.

Analysis of Peripheral T-Lymphocyte Subpopulations in Patients with Early Syphilis by Flow Cytometry

Objective: To study the cellular immunity function of patientswith early syphilis and the effects on immune modifiersEsberitox N or IFN. Methods: T-lymphocyte subpopulations of the peripheralblood in 44 patients with syphilis and 40 healthy controls wereexamined by flow cytometry. Results: The number of CD_4+ cells and the CD_4+/CD_8+ ratioin patients with syphilis were found to be significantly lowerthan those in the control (P<0.0l), while the number of CD_8+cells was higher than that in the control (P<0.01). TheCD_4+/CD_8+ ratio in those with active disease was lower thanthat in those who had been cured (P<0.05). The CD_4+ countand the CD_4+/CD_8+ ratio in those treated with antibiotics alone(Penicillin G or Cephalosporins) were lower than those treatedwith both antibiotics and immunomodulators (P<0.05). Conclusions: Cellular immunity in the patients with earlysyphilis was prominently suppressed, and treatment withimmunomodulators may be helpful for the recovery of cellularimmunity of these patients.

Effect of copper excess on peripheral blood T-lymphocytes in the chicken

Experimental study was conducted to examine the effect of copper excess on the peripheral blood T-lymphocyte by the methods of flow cytometry （FCM） and experimental pathology. 420 one-day-old Avian chickens were randomly divided into seven groups, and fed on diets as follows： 1. controls （Cu 11mg/kg） and 2. copper excess （ Cu 100mg/kg, copper excess group Ⅰ; Cu 200mg/kg, copper excess group Ⅱ; Cu 30（hng/kg, copper excess group Ⅲ; Cu 400mg/kg, copper excess group Ⅳ; Cu 500mg/kg, copper excess group Ⅴ; Cu 600mg/kg, copper excess group VI） for six weeks. The results were as follows- 1 ） In thymus, lymphocytes in the medulla were decreased in number in copper excess groups Ⅲ,Ⅳ,Ⅴ, and Ⅵ, and the increased and enlarged thymic corpuscles and the proliferated reticular cells were also observed in both copper excess group V and copper excess group VI in comparison with those of control group. 2） The percentage of CD4＋ T cells was markedly decreased from 2 to 6 weeks of age in copper excess groups Ⅳ, Ⅴ and Ⅵ （P 〈0.05 or P 〈0.01）. 3） The percentage of CD8＋ T cell was not varied in six copper excess groups during the experiment when compared with that of control group （ P 〉 0.05 ）. 4） The CD4 ＋ / CD8＋ ratio was lower from 2 to 6 weeks of age in copper excess groups Ⅳ, Ⅴ and VI than in control group （ P 〈0.05 or P 〈0.01）. 5） It was concluded that dietary copper in excess of 300mg / kg suppressed the development of T-lymphocytes and reduced the percentage of CD4＋ T cells and the CD4＋/CD8＋ ratio, and resulted in pathological injury of the thymus. Cellular immune function was finally impaired.

T Lymphocytes and Th1/Th2 Cytokines in Peripheral Blood of Patients with Recurrent Genital Herpes

Objective: This study analyzed the T lymphocytes and Th1/Th2 type cytokine profile shift in the peripheral blood ofpatients with recurrent genital herpes (RGH). Methods: Immunofluorescent staining or cell surface antigenand intracellular cytokines(IL-2、IL-4、IL-12、IFN-r)inperipheral blood from 20 RGH patients and 10 controls wereanalyzed using flow cytometric techniques. Results: RGH patients had signiflcantly lower levels ofCD3^+T cells, CD4^+T cells and CD4^+ T/ CD8^+ T cells ratiocompared to control levels (P<0.001), and IL-2-producing,IFN-r-producing and IL-12-producing T cells were increasedin RGH patients (CD4^+T: P<0.001, CD8^+T: P<0.05respectively), whereas IL-4-producing T cells were increased inRGH patients compared to controls (CD4^+T: P<0.05; CD8^+T:P<0.001 respectively). Conclusions: RGH patients have T lymphocyte subsetvariations and Th1/Th2 cytokine changes. The increase in Th2cells Th1/Th2 imbalance may have important implications forRGH pathogenesis.

Effect of Xuebijing injection on peripheral T-lymphocyte subpopulations in patients with severe trauma

Objective： To investigate the effect and clinical significance of Xuebijing injection on peripheral T-lymphocyte subpopulations in patients with severe trauma. Methods： Thirty-three patients with severe trauma were randomly divided into a control group （n=16） and a treatment group （n=17）. The patients of two groups were all treated conventionally, and the only difference was that Xuebijing injection was given to patients of the treatment group. The CD4^＋ and CD8^＋ subpopulations of T-lymphocyte in the peripheral blood were detected respectively on admission, 3rd and 5th days after trauma by double antibody labeling and flow cytometry. Results： The CD4^＋ T-lymphocytes and CD4^＋/CD8^＋ ratio of peripheral blood in patients with severe trauma decreased markedly on the 3rd and 5th days after trauma. Furthermore, compared with control group, the peripheral CD4^＋ T-lymphocytes and CD4^＋/CD8^＋ ratio of treatment group renewed obviously on the 5th day after trauma, and showed statistical differences （P〈0.05）. Conelusion： In the treatment of patients with severe trauma, the early use of Xuebijing injection is effective in correcting disorder or suppression of T-lymphocyte subpopulations regulating network, and promoting a more balanced profile of immunologic function.

Protein kinase C-δ and -β coordinate flow-induced directionality and deformation of migratory human blood T-lymphocytes

T-tym phocyte migration under flow is critical for immune responses, but the mechanisms by which flow modulates the migratory beha- viors of T-lymphocytes remain unclear. Human peripheral blood T-lymphocytes （PBTLs）, when stimulated with phorboL 12-myristate 13-acetate （PMA）, stretched their ceU bodies dramatically and moved alongthe flow direction. In contrast, stromal ceil-derived factor- lα-stimulated PBTI.s deformed and migrated in a random manner. Here we elucidated the molecular mechanisms underlying flow- induced directionality and deformation of PMA-stimulated PBTLs. PMA primed PBTLs for polarization under flow, with protein kinase C （PKC）-δ enriched in the leading edge, PKC-β1 in the microtubuie organizing center, and PKC-1311 in the uropod and peripheral region. PKC-δ regulated cell protrusions in the leading edge through Tiaml/Racl/caLmoduUn, whereas PKC-β regulated RhoA/Rho- associated kinase activity and microtubule stability to modulate uropod contractility and detachment. Our findings indicate that PKC-δ and -β coordinate in the cell Leading edge and uropod, respectively, to modu｜ate the directionality and deformability of migratory T-Lymphocytes under flow.

Effect of cold-dryness on pulmonary and immunologic function in chronic obstructive pulmonary disease model rats

OBJECTIVE： To study the effects of cold-dryness on pulmonary and immunologic function of peripheral T-lymphocytes in chronic obstructive pulmonary disease （COPD） model rats, and to provide references for the prevention and treatment of cold-dryness COPD in the Xinjiang region. METHODS： The COPD model was established with an elastase drip into the trachea combined with smoking. The cold-dryness COPD model was developed by stressing with a cold-dry environment. Success of the model was determined by observation of pathologic lung sections. Rats were sacrificed by exsanguination from the femoral artery and changes of peripheral blood CD4＋, CD8＋, and CD4＋/CD8＋ were detected by flow cytometryo Data were analyzed with SAS 11.5 statistical software. RESULTS： On the ninetieth day after ending the ex- periment, Peak expiratory flow in the cold-dryness COPD group was lower than that in the COPD and normal control groups （P〈0.01）. The time of inspiration in the cold-dryness COPD group was higher than that in the COPD and normal groups （P〈0.05）. Time of expiration （Te） in the cold-dryness COPD group was higher than that in the COPD and normal groups （P〈0.01）. 50% tidal volume expiratory flow （EFS0） in the cold-dryness COPD group was lower than that in the COPD and normal groups （P〈 0.01）, and EFS0 in the COPD group was lower than that in the normal group （P〈0.05）. CD4＋ content of peripheral blood in the cold-dryness COPD group was lower than that in the COPD and the normal groups （P〈0.05）. CD8＋ content in the cold-dryness COPD and COPD groups was higher than that in the normal control group （P〈0.01）, and CD8＋ content in the cold-dryness COPD group was higher than that in the COPD group （P〈0.01）. CD4＋/CD8＋ in the cold-dryness COPD group and the COPD group was lower than that in the normal control group （P〈0.01）, and CD4＋/CD8＋ in the cold-dryness COPD group was lower than that in the COPD group （P〈0.05）. CONCLUSION： In the cold-dryness COPD model, CD8＋ increased and CD4＋/CD8＋ decreased. Moreover, cold-dryness may aggravate this state. The effects of cold-dryness on pulmonary function main- ly manifested as prolongation of Te and decrease of EF50, which could be one of causes of cold-dryness environment in the northwest of China leading to COPD with region characteristics.